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Abstract 摘要

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The first-in-class soluble guanylate cyclase (sGC) stimulator riociguat was recently introduced as a novel treatment option for pulmonary hypertension. Despite its outstanding pharmacological profile, application of riociguat in other cardiovascular indications is limited by its short half-life, necessitating a three times daily dosing regimen. In our efforts to further optimize the compound class, we have uncovered interesting structure–activity relationships and were able to decrease oxidative metabolism significantly. These studies resulting in the discovery of once daily sGC stimulator vericiguat (compound 24, BAY 1021189), currently in phase 3 trials for chronic heart failure, are now reported.
首次引入的同类可溶性鸟苷酸环化酶（sGC）激动剂瑞伐西单抗最近被作为一种新型肺动脉高压治疗选择。尽管其药理特性卓越，但由于其半衰期短，瑞伐西单抗在其他心血管适应症中的应用受到限制，需要每日三次的给药方案。在我们进一步优化该化合物类别的努力中，我们发现了有趣的构效关系，并显著降低了氧化代谢。这些研究导致了每日一次 sGC 激动剂维西单抗（化合物 24，BAY 1021189）的发现，目前该药物正在接受慢性心力衰竭的 3 期临床试验，现在予以报道。

Subjects

Introduction 引言

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The NO–sGC–cGMP axis belongs to the key signal transduction pathways involved in regulating the cardiovascular system. (1, 2) Central to this pathway is soluble guanylate cyclase (sGC), an intracellular enzyme present in the smooth muscle cells of blood vessels and in platelets but also in various other cell types like cardiomyocytes. sGC displays a high affinity for, and is activated by, the first messenger signaling molecule nitric oxide (NO). Biosynthesis of NO is mediated by three distinct isoforms of nitric oxide synthases, endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), and the inducible isoform of nitric oxide synthase (iNOS), all synthesizing NO from l-arginine. Within endothelial cells, NO is produced by eNOS and diffuses rapidly to the underlying smooth muscle cells where it binds to its target enzyme sGC. This binding leads to the stimulation of sGC, resulting in increased production of intracellular second messenger cGMP. cGMP interacts with three types of intracellular proteins such as cGMP-dependent protein kinases, cGMP-regulated ion channels, and phosphodiesterases. Further downstream, these transduction cascades mediate various physiological and tissue-protective effects including smooth muscle relaxation and inhibition of smooth muscle proliferation, leukocyte recruitment, and platelet function. The pathogenesis of various diseases, especially those of the cardiovascular system, has been related to insufficient bioavailability of NO and thus impaired stimulation of sGC and therefore decreased cGMP production. (3-7)
NO-sGC-cGMP 通路是调节心血管系统的重要信号转导通路之一。（1，2）该通路的核心是可溶性鸟苷酸环化酶（sGC），这是一种存在于血管平滑肌细胞和血小板中的细胞内酶，也存在于各种其他细胞类型中，如心肌细胞。sGC 对第一信使信号分子一氧化氮（NO）具有高亲和力，并由其激活。一氧化氮的生物合成由三种不同的氧化酶同型异构体介导，即内皮型一氧化氮合酶（eNOS）、神经元型一氧化氮合酶（nNOS）和诱导型一氧化氮合酶（iNOS），它们均从 L-精氨酸合成一氧化氮。在内皮细胞中，一氧化氮由 eNOS 产生并迅速扩散到其下方的平滑肌细胞，在那里它与靶酶 sGC 结合。这种结合导致 sGC 的刺激，从而增加细胞内第二信使 cGMP 的产生。cGMP 与三种类型的细胞内蛋白相互作用，如 cGMP 依赖性蛋白激酶、cGMP 调节的离子通道和磷酸二酯酶。下游进一步，这些转导级联介导多种生理和组织保护作用，包括平滑肌松弛、抑制平滑肌增殖、白细胞募集和血小板功能。各种疾病的发病机制，尤其是心血管系统疾病，与一氧化氮的生物利用度不足有关，从而损害 sGC 的刺激，因此导致 cGMP 产生减少。 (3-7)

sGC itself is a cytosolic, heterodimeric protein composed of an α- and a β-subunit with a prosthetic heme group located in the β-subunit (heme-binding domain). (8-13) Mechanistically, NO stimulates sGC by binding to the Fe²⁺ of the heme group, which induces cleavage of an Fe²⁺–histidine (His¹⁰⁵) bond, likely resulting in conformational reorganizations which propagate into the catalytic subunit, increasing cGMP production. cGMP interacts with three types of intracellular proteins, namely cGMP-dependent protein kinases, cGMP-regulated ion channels, and phosphodiesterases. Further downstream, these transduction cascades mediate various physiological and tissue-protective effects, including smooth muscle relaxation and inhibition of smooth muscle proliferation, leukocyte recruitment, and platelet function. (1)
sGC 本身是一种细胞质中的异源二聚体蛋白，由 α-亚基和 β-亚基组成，β-亚基中含有一个位于其上的辅因子血红素（血红素结合域）。（8-13）从机制上讲，NO 通过与血红素中的 Fe2+ 结合来刺激 sGC，这会导致 Fe2+-组氨酸（His105）键的断裂，这很可能是导致构象重组，进而传播到催化亚基，增加 cGMP 的产生。cGMP 与三种类型的细胞内蛋白相互作用，即 cGMP 依赖性蛋白激酶、cGMP 调节的离子通道和磷酸二酯酶。进一步下游，这些转导级联反应介导各种生理和器官保护作用，包括平滑肌松弛、抑制平滑肌增殖、白细胞募集和血小板功能。（1）

sGC can exist in two different states, a native heme-containing or reduced form of sGC, which is the endogenous receptor for NO, and a heme-free form of sGC. (1, 14) Under conditions of oxidative stress, which is thought to be causal in the pathogenesis of many cardiovascular diseases, reactive oxygen species (ROS) are produced. ROS are capable of oxidizing the heme iron of sGC (Fe²⁺ → Fe³⁺), resulting ultimately in heme loss from oxidized sGC. As a consequence, the heme-free form of sGC is no longer binding NO, is not responsive to NO, and thus termed dysfunctional. During the last 20 years, two distinct compound classes have been discovered at Bayer that are capable of activating sGC in a NO-independent manner, the heme-dependent sGC stimulators and the heme-independent sGC activators. sGC stimulators display a dual mode of action: they synergize with endogenous NO and, on top of this, are also able to directly stimulate the native form of the enzyme independently of NO, resulting in increased cGMP production. (15) In contrast, sGC activators are capable of activating the dysfunctional heme-free sGC, resulting in increased cGMP production even under conditions of reduced NO bioavailability. (16)
sGC 可以存在于两种不同的状态，即含有天然血红素的 sGC 或还原形式的 sGC，它是 NO 的内源性受体，以及不含血红素的 sGC。在氧化应激条件下，氧化应激被认为是许多心血管疾病发病机制的原因，会产生活性氧（ROS）。ROS 能够氧化 sGC 中的血红素铁（Fe2+ → Fe3+），最终导致氧化 sGC 失去血红素。因此，不含血红素的 sGC 不再结合 NO，对 NO 无反应，因此被称为功能障碍。在过去 20 年里，拜耳公司发现了两种不同的化合物类别，它们能够以 NO 非依赖性方式激活 sGC，即血红素依赖性 sGC 激动剂和血红素非依赖性 sGC 激活剂。sGC 激动剂具有双重作用模式：它们与内源性 NO 协同作用，并且在此基础上，还能直接刺激酶的天然形式，独立于 NO，从而增加 cGMP 的产生。 (15) 相比之下，sGC 激活剂能够激活功能失调的无铁血红素 sGC，即使在 NO 生物利用度降低的条件下，也能增加 cGMP 的产生。 (16)

The discovery of the sGC stimulators at Bayer, along with the efforts of several other pharmaceutical companies to identify sGC stimulators and activators, has recently been reviewed. (2) Riociguat (1) is the first sGC stimulator to have made a successful transition from animal experiments to controlled clinical studies in patients. (17) In 2013, 1 gained market approval for two life-threatening diseases: pulmonary arterial hypertension (PAH) (18) and chronic thromboembolic pulmonary hypertension (CTEPH). (19, 20) However, application of 1 in other cardiovascular indications, such as heart failure, is limited by its short half-life. (21)
贝耶尔公司发现了 sGC 激动剂，以及多家制药公司努力寻找 sGC 激动剂和激活剂的努力，最近得到了综述。 (2) Riociguat（1）是第一个成功从动物实验过渡到患者临床试验的 sGC 激动剂。 (17) 2013 年，1 号药物获得了市场批准，用于治疗两种危及生命的疾病：肺动脉高压（PAH）(18) 和慢性血栓栓塞性肺动脉高压（CTEPH）(19, 20)。然而，由于 1 号药物的半衰期较短，其在其他心血管适应症，如心力衰竭中的应用受到限制。 (21)

Herein, we report our recent efforts in optimizing the pharmacokinetic profile of sGC stimulators, culminating in the discovery of vericiguat (compound 24, BAY 1021189), which has been evaluated in phase 2 studies (SOCRATES trials) in heart failure patients with reduced (HFrEF) and preserved (HFpEF) ejection fraction. (22, 23) Currently, 24 is being investigated in a phase 3 outcome study (VICTORIA trial) in heart failure patients with HFrEF in codevelopment with MSD (vericiguat code name: MK-1242). (24) That study is aimed at assessing whether 24, on top of standard of care, is able to decrease mortality and morbidity in such HFrEF patients.

Results and Discussion

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Optimization Strategy

Our goal was to identify orally bioavailable sGC stimulators with a longer duration of action than 1 in order to support a profile allowing for a once daily oral dosing and less oxidative metabolism in order to lower interaction potentials. 1 is a very potent sGC stimulator in vitro and in vivo; however, it has a moderate half-life in different animal species (25) and this pharmacokinetic profile translated into a three times daily dosing regimen in patients. N-Demethylation to compound 9, as described by Gnoth et al. (26) in 2015, is the major biotransformation of 1 and is mainly catalyzed by CYP1A1 and also by CYP3A4, CYP3A5, and CYP2J2. (21, 26-31) Our strategy was to further optimize the metabolic stability of 1 and hence reduce blood clearance to achieve a longer half-life. We began by optimizing the substituents on the 5-carbamoyl residue on the pyrimidine ring, aiming to achieve metabolically more stable derivatives while maintaining good potency. Later, in a second optimization step, we focused on variations of the central pyrazolopyridine scaffold.

Chemistry

Scheme 1 outlines the strategy for the synthesis of the 5-carbamoyl variations. Starting from triamine 1c, which is available in two steps from amidine 1a and phenyldiazenyl-substituted malonodinitrile 1b, selective carbamate formation at the 5-amino group was achieved either by treatment with the corresponding chloroformate or by addition of the respective alcohol preactivated with triphosgene to yield compounds 9–14. (25) Deprotonation of the carbamate NH group and subsequent treatment with the corresponding halide or trichloromethanesulfonate electrophile allowed further derivatization to the corresponding N-alkylated carbamates (1, 2, 4–8). Alternatively, the alkyl side chain on the 5-amino group could first be introduced via reductive amination and the product 1d subsequently treated with methyl chloroformate, giving access to the hydroxyethyl derivative 3. For the cyclic 5-(2-oxo-1,3-oxazolidin-3-yl) analogues 15–18, the previously employed conditions for the selective formation of carbamates provided 1e–g bearing a chloro- or bromo-substituted alkyl chain, allowing for a base-induced intramolecular cyclization (NaHMDS).

The synthesis of the compounds with a core variation is outlined in Schemes 2–5. For the 1H-pyrazolo[4,3-b]pyridine derivative 19, the synthesis started from 3,5-dichloropyridine-2-carbonyl chloride (2a), which was converted into 3-(2-fluorobenzyl)-1H-pyrazolo[4,3-b]pyridine (2b) in four steps. The fluoro derivative 2d was synthesized in a similar manner from the corresponding difluoropyridine analogue 2c. Both key intermediates 2b and 2d were then reacted with 2-chloro-5-nitropyrimidine-4,6-diamine, (32) followed by reduction of the pyrimidine 5-nitro group and methyl carbamate formation, to give the 1H-pyrazolo[4,3-b]pyridine core analogues 19 and 20 (Scheme 2).

Scheme 2. Synthesis of Compounds 19 and 20^a

A different route was used for the preparation of 1H-pyrazolo[3,4-c]pyridazine 21 (Scheme 3), which began with the cyclization of methyl 2-aminothiophene-3-carboxylate (3a) with hydrazine hydrate to give 1H-pyrazolo[3,4-c]pyridazin-3-ol (3b). (33) Subsequent bromination afforded intermediate 3c, which was alkylated with 2-fluorobenzyl bromide to yield 3d. Then, conversion of the bromide into the cyanide 3e and Pinner-type reaction of the cyano group with sodium methoxide and ammonium chloride afforded key intermediate 3f. Further modifications, analogous to the formation of carbamate 9 from 1a (Scheme 1), yielded the methyl carbamate derivative 21.

The preparation of imidazo[1,5-a]pyrimidine 22 (Scheme 4) started with condensation of the readily available thioimide 4a and amine 4b to access aminoimidazole 4c. Further orthoester condensation then gave the bicyclic imidazopyrimidine derivative 4d bearing an ethyl ester functionality. Subsequent saponification, amidation, and dehydration yielded nitrile 4g as a key intermediate. Pinner-type reaction with sodium methoxide and ammonium chloride delivered the corresponding amidine 4h, which could be converted into the final compound 22 in analogy to the previous examples.

The synthesis of the related imidazo[1,5-b]pyridazine derivative 23 (Scheme 5) started from methyl 5-aminolevulinate hydrochloride (5a). Addition of hydrazine hydrate yielded the dihydropyridazinone 5b, which was reacted with the readily available acid chloride 5c under basic conditions. The resulting amide was oxidized with bromine to provide the corresponding pyridazinone 5d, which underwent an intramolecular condensation in the presence of phosphoryl chloride to give the bicyclic intermediate 5e. Dechlorination of the pyridazine core catalyzed by palladium on charcoal and subsequent bromination of the imidazo core with N-bromosuccinimide gave access to 5f. Further modifications, as outlined previously, then yielded amidine 5g as a key intermediate which was converted into the final target compound 23.

The synthesis of the 1H-pyrazolo[3,4-b]pyridine derivative (24) (Scheme 6), involved initial activation of 2,2,3,3-tetrafluoro-1-propanol (6a) with trifluoromethanesulfonic anhydride and treatment with morpholine to yield 6b after aqueous workup and distillation. At 135 °C, 6b was alkylated with methylmethanesulfonate, leading to quaternized derivative 6c which was converted into 6d using sodium hydroxide. Further reaction with morpholine and triethylamine yielded 6e after crystallization from toluene. Acrylaldehyde derivative 6e was reacted with ethyl 5-amino-1-(2-fluorobenzyl)-1H-pyrazole-3-carboxylate (25) in ethanol, yielding ethyl ester intermediate 6f. Ester 6f was then converted in three steps into the corresponding amidine 6i via amide 6g and nitrile 6h in analogy to the chemistry outlined in Scheme 4. The synthesis of 24 was completed as described for compound 9 (Scheme 1) via the diazenyl derivative 6j and triamine 6k.

SAR and DMPK Optimization

The SAR of the series of novel N-substituted methyl carbamates 1–8 was explored using a cGMP formation assay with sGC-overexpressing Chinese hamster ovary (CHO) cells. (34, 35) In addition, the metabolic stability of the compounds was assessed in vitro by incubation with rat hepatocytes. Initially, different carbamate N-substituents were tested with the aim of achieving higher metabolic stability than 1 while maintaining high potency (Table 1). Increasing steric bulk as in the ethyl derivative 2 had no beneficial effect on either potency or metabolic stability. The introduction of polarity (e.g., a hydroxyethyl functionality, as in 3) led to a significant loss of potency relative to 1, along with decreased metabolic stability. The next strategy, the introduction of fluorine atoms at the terminal position of the N-substituent, was designed to block metabolism. Interestingly, both the 2,2-difluoroethyl and 2,2,2-trifluoroethyl derivatives, 4 and 5, exhibited high potency in the cGMP assay but had a 2–4.5-fold higher clearance in rat hepatocytes. N-Fluorobenzyl substitution revealed that the ortho-fluoro derivative 6 is significantly more potent (MEC = 0.1 μM) than the meta and para isomers, 7 and 8 (MEC = 0.2 and 0.7 μM, respectively). Nevertheless, all the benzylic derivatives had a high clearance in rat hepatocytes. These results and the testing of many further derivatives (data not shown) suggested that improving the metabolic stability by altering the N-substitution might be very difficult to achieve. Hence, optimization efforts were focused on the main metabolite of 1, the N-desmethyl derivative 9. This compound is less potent than 1 but displayed a somewhat higher metabolic stability in the rat hepatocyte assay, with a clearance of 0.1 L/h/kg. When tested in vivo in rats (iv dosing), compound 9 had a moderate clearance of 1.2 L/h/kg and a short half-life of only 1.2 h.

Table 1. Properties of the N-Substituted Methyl Carbamates 1–8

^{Table a}

MEC: Minimal effective concentration to achieve stimulation of cGMP formation (≥3-fold increase in basal luminescence) in a recombinant sGC-overexpressing cell line. (34)

Therefore, variations in the carbamate alkoxy group while leaving the N-position unsubstituted were examined next, with the goal of improving the half-life and other properties relative to compound 9 (Table 2).

Table 2. Properties of the N–H Alkyl Carbamates 9–14

^{Table a}

MEC: Minimal effective concentration to achieve stimulation of cGMP formation (≥3-fold increase in basal luminescence) in a recombinant sGC-overexpressing cell line. (34)

^{Table b}

n.d.: not determined.

The introduction of steric bulk in the form of an isopropyl residue (compound 10) led to a strong decrease in potency relative to 9 as well as decreased metabolic stability. Interestingly, there was increased potency with cyclobutyl derivative 11; however, 11 was also unfavorable with respect to rat hepatocyte clearance. The oxetanyl group is well-known in medicinal chemistry for improving metabolic stability (36, 37) over related alkyl derivatives. Remarkably, the exchange of cyclobutyl for oxetanyl (compound 12) led to a 14-fold higher stability in rat hepatocytes, but sGC stimulation was weaker than with derivative 9 or 11. Because the permeability across Caco-2 cell monolayers was also very low (P_app A–B = 2 nm/s), combined with a high efflux ratio of 74, compound 12 was not pursued further. Additional efforts to improve the overall profile by the introduction of fluorine or steric bulk (compounds 13 and 14) led to a slight increase in potency (MEC = 0.2 vs 0.3 μM for 9); however, metabolic stability was dramatically reduced.

The oxazolidinone derivatives 15–18, as conformationally fixed versions of the foregoing carbamates, were also studied to assess the influence of the constrained nature on sGC stimulation (Table 3). The parent oxazolidinone 15 was as equipotent as carbamate 9 and exhibited slightly decreased stability in rat hepatocytes. Intravenous dosing of compound 15 to rats resulted in a moderate clearance of 1.9 L/h/kg and a short half-life of about 1.0 h, and thus 15 was not profiled further. Other attempts to increase metabolic stability by the introduction of steric bulk were unsuccessful, probably due to the more lipophilic (38) character of the resulting compounds 16–18.

Table 3. Properties of the Oxazolidinones 15–18

^{Table a}

MEC: Minimal effective concentration to achieve stimulation of cGMP formation (≥3-fold increase in basal luminescence) in a recombinant sGC-overexpressing cell line. (34)

^{Table b}

n.d.: not determined.

In summary, although further optimization of the carbamate motif of 1 could not be achieved, a number of interesting and novel SAR observations were made, with the N-unsubstituted carbamate 9 being identified as the most promising path forward. Thus, the focus of the optimization strategy was shifted away from the carbamate moiety and our efforts were directed to the central scaffold and the identification of alternative cores that could lead to superior overall pharmacokinetic profiles of the corresponding compounds.

Our studies with respect to central scaffold modifications are summarized in Table 4. Efforts were concentrated on novel core systems which had not been tested before in combination with 4,6-diaminopyrimidin-5-ylcarbamates.

Table 4. Properties of the Core Variation Compounds 19–24

^{Table a}

MEC: Minimal effective concentration to achieve stimulation of cGMP formation (≥3-fold increase in basal luminescence) in a recombinant sGC-overexpressing cell line. (34)

^{Table b}

n.d.: not determined.

First, changes were made to the pyrazolo portion of the molecule, leading to the 1H-pyrazolo[4,3-b]pyridine derivative 19. In addition to the benefit of a shorter synthetic route than that for compound 9, 19 proved to be a moderately potent sGC stimulator with an MEC of 1.2 μM and had high metabolic stability when tested in rat hepatocytes. The introduction of substituents at the 6-position of the 1H-pyrazolo[4,3-b]pyridine core was, in general, not well tolerated, leading to a dramatic loss of potency, with the exception of the 6-fluoro derivative 20, which exhibited an MEC of 0.5 μM and good metabolic stability. To our surprise, compounds 19 and 20 had very different in vivo clearances when compared in a rat pharmacokinetic experiment (iv dosing): fluoro derivative 20 had a low clearance of 0.3 vs 1.0 L/h/kg for compound 19. As metabolite identification did not point to metabolism occurring at the pyridine core, the rationale for this 3-fold reduction in blood clearance remains unclear.

Novel 1H-pyrazolo[3,4-c]pyridazine 21 proved to be a reasonably potent sGC stimulator but exhibited a high clearance of 3.8 L/h/kg when tested in vivo in rats and thus was not further profiled, along with imidazo[1,5-a]pyrimidine 22 for similar reasons. In contrast, imidazo[1,5-b]pyridazine 23 exhibited potent sGC stimulator properties (MEC = 0.7 μM), good metabolic stability in rat hepatocytes, and a low to moderate clearance of 0.9 L/h/kg after intravenous dosing to rats. Finally, revisiting 1H-pyrazolo[3,4-b]pyridine 9 with an additional fluorine at the 5-position resulted in the potent sGC stimulator 24 (MEC = 0.3 μM) with good metabolic stability in rat hepatocytes and a surprisingly low clearance of 0.3 L/h/kg after intravenous dosing to rats (Table 4). Thus, the blood clearance of derivative 9 in rats (1.2 L/h/kg) was reduced 4-fold by fluorination at position 5.

Finally, compounds 20, 23, and 24 were selected for further pharmacokinetic profiling across species (Table 5). Fluoropyrazolo[3,4-b]pyridine derivative 24 exhibited the best overall pharmacokinetic profile by far, with a low clearance and long half-life in rats and dogs after intravenous dosing, as well as high oral bioavailability. In addition, 24 had no inhibitory effects on major CYP isoforms (1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, and 3A4), as indicated by IC₅₀ values of >50 μM. Metabolite profiling in human hepatocytes was characterized by a low turnover, with glucuronide 25 (major) and debenzylated compound 26 (minor) being the only metabolites identified (Figure 1). Thus, the main biotransformation pathway shifted from predominantly phase I oxidative CYP-mediated metabolism (1) to primarily phase II UGT-mediated conjugation with glucuronic acid.

Table 5. In Vivo Pharmacokinetic Properties of 20, 23, and 24 in Comparison with 1

compd	species	V_ss [L/kg]	CL_b [L/h/kg]	t_1/2 [h]	bioavailability [%]
20	rat	0.5	0.3	1.5	26
	dog	1.0	0.2	4.1	56
23	rat	0.3	0.9	0.5	37
	dog	2.0	0.9 (Cl_p)^a	1.8	n.d.^b
24	rat	1.0	0.3	3.4	65
	dog	1.4	0.2	6.2	75
1	rat	1.2	1.3	1.4	46
	dog	0.7	0.3 (Cl_p)^a	2.4	79

Cl_p: plasma clearance.

n.d.: not determined.

Figure 1. Biotransformation products of 24 in human hepatocytes.

Pharmacology

24 was extensively profiled preclinically in vitro, on the isolated sGC enzyme and on isolated vessels, ex vivo in isolated hearts, and in vivo in a rat model of cardiovascular disease associated with cardiorenal syndrome.

Highly Purified Recombinant sGC

Studies on the in vitro effects of 24 on highly purified sGC revealed that 24 (0.01 μM to 100 μM) stimulates recombinant sGC concentration dependently, by 1.7-fold to 57.6-fold (Figure 2). When combined with the NO donor diethylamine/nitric oxide complex (DEA/NO), 24 and DEA/NO had a synergistic effect on the enzyme activity over a wide range of concentrations. At highest concentrations of 24 (100 μM) and DEA/NO (100 nM), the specific activity of sGC was 341.6-fold above baseline (Figure 2). Moreover, the effects of 24 in the presence of the sGC inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) were evaluated. The sGC stimulation induced by 24 was nearly completely blocked by ODQ and reduced to 18-fold (Figure 2). Thus, 24 exhibits all the characteristics of a potent and selective sGC stimulator, stimulating sGC NO-independently and in synergy with NO. In addition, 24 predominantly acts on the heme-containing nonoxidized form of sGC.

Figure 2. Effects of 24 and NO on the stimulation of highly purified sGC and blocking effects of the sGC inhibitor ODQ.

sGC-Overexpressing Cells

The stimulation of sGC by 24 was examined with a recombinant CHO cell line overexpressing rat sGC. 24 stimulated the sGC reporter cell line concentration dependently, with an EC₅₀ of 1005 ± 145 nM. In the presence of the NO donor S-nitroso-N-acetyl-d,l-penicillamine (SNAP) (30 and 100 nM), the EC₅₀ value shifted to 39.0 ± 5.1 and 10.6 ± 1.7 nM, respectively. In the presence of ODQ, pretreatment of the sGC reporter cell line with 10 μM ODQ for 3 h resulted in a significantly reduced efficacy of 24, with an EC₅₀ of 256 ± 40 nM being observed.

Isolated Vessels and Tolerance

24 inhibited phenylephrine-induced contractions of rabbit saphenous artery rings, rabbit aortic rings, and canine femoral vein rings concentration dependently, with IC₅₀ values of 798, 692, and 3072 nM, respectively. In addition, 24 inhibited the U46619-induced contractions of porcine coronary artery rings concentration dependently, with an IC₅₀ of 956 nM.

Chronic administration of organic nitrates leads to the rapid development of nitrate tolerance. Thus, the vasorelaxant effect of 24 on isolated saphenous artery rings taken from normal and nitrate-tolerant rabbits was examined. Treatment with isosorbide dinitrate for 3–4 days resulted in a marked inhibition of glycerol trinitrate (GTN) mediated vasodilation. GTN inhibited phenylephrine-induced contractions with an IC₅₀ of 1.9 nM in control vessels and an IC₅₀ of 9.6 nM in tolerant vessels, confirming the presence of nitrate tolerance. In contrast to GTN, 24 is a potent inhibitor of phenylephrine-induced contractions both in normal and nitrate-tolerant saphenous artery rings, with IC₅₀ values of 5.6 and 5.8 nM, respectively.

Langendorff-Perfused Hearts

In rat heart Langendorff preparations, ex vivo, 24 reduced the coronary perfusion pressure in a concentration-dependent manner (Figure 3). Up to the highest concentration tested, 24 had no effect on heart rate, left ventricular diastolic pressure, and contractility.

Figure 3. Effects of 24 on rat heart Langendorff preparations.

Long-Term Study with L-NAME-Treated Renin Transgenic Rats

The potential impact on cardiovascular health of the stimulation of sGC was evaluated by determining the long-term effects of 24 on hemodynamic and hormonal parameters in renin transgenic rats (RenTG) carrying the additional mouse renin gene (mRenR2)27. These RenTG(mRenR2)27 rats were additionally treated with the NO synthase inhibitor N_ω-nitro-l-arginine methyl ester (L-NAME) in the drinking water. This well-established rodent disease model is characterized by hypertension, heart and kidney failure, and increased mortality. (39) The RenTG(mRenR2)27/L-NAME-supplemented rats were chronically treated with either placebo (2 mL/kg qd), which served as the control group, or 24 (3 or 10 mg/kg qd).

Effects on Blood Pressure and on the Heart

Chronic oral treatment with 3 or 10 mg/kg 24 qd resulted in a significant attenuation of blood pressure increase during the course of the study. However, the overall rise of blood pressure increase was not halted in the 3 and 10 mg/kg treatment groups (Figure 4).

Figure 4. Increase in systolic blood pressure in mmHg during the course of the study with L-NAME-treated renin transgenic rats.

In addition, a significant and dose-dependent reduction of heart hypertrophy, in both the right and left ventricle, was found in the 3 and 10 mg/kg treatment groups compared to the placebo group (Figure 5). Furthermore, plasma ANP levels decreased significantly in both treatment groups, also suggesting a functional improvement of the heart.

Figure 5. Right and left ventricle weight normalized on body weight (left/middle) and plasma atrial natriuretic peptide levels [in pg/mL] at the study end, after 3 weeks of treatment.
图 5. 右心室和左心室重量按体重标准化（左/中）以及研究结束时，经过 3 周治疗后血浆心房利钠肽水平[以 pg/mL 计]。

Effects on the Kidneys 肾脏影响

With respect to kidney damage, 24 treatment at 3 or 10 mg/kg led to a significant reduction in kidney injury molecule Kim-1 and osteopontin expression which are used as biomarkers for renal injury and dysfunction (data not shown). In addition, proteinuria was significantly and dose dependently decreased in the treatment groups, also suggesting a functional improvement of the kidneys (Figure 6).
关于肾脏损伤，3 毫克/千克或 10 毫克/千克的 24 小时治疗显著降低了肾脏损伤分子 Kim-1 和骨桥蛋白的表达，这些指标被用作肾损伤和功能障碍的生物标志物（数据未显示）。此外，治疗组的蛋白尿显著且剂量依赖性地减少，这也表明肾脏功能有所改善（图 6）。

Figure 6. Effects on proteinuria at the study end after 3 weeks of treatment.
图 6. 治疗结束后 3 周对蛋白尿的影响。

Effects on Mortality 死亡率影响

Treatment with 24 resulted in a significant and dose-dependent increase in survival rates. In the 3 and 10 mg/kg qd treatment groups, the rat survival rate was 70% and 90%, respectively, at the study end. In contrast, the survival rate in the placebo group was only 25% after 21 days (Figure 7).
治疗 24 小时后，生存率显著且呈剂量依赖性增加。在 3 毫克/千克和 10 毫克/千克每日一次的治疗组中，研究结束时大鼠的生存率分别为 70%和 90%。相比之下，安慰剂组的生存率在 21 天后仅为 25%（见图 7）。

Figure 7. Kaplan–Meier survival curves.
图 7. Kaplan-Meier 生存曲线。

These in vivo data strongly suggest that the sGC stimulator 24 can maintain heart and kidney function in a model of hypertension-induced end-organ damage, with substantially reduced overall mortality, strongly suggesting a beneficial role of 24 for the treatment of cardiovascular diseases associated with cardiorenal syndrome. (40)
这些体内数据强烈表明，sGC 激动剂 24 可以在高血压诱导的终末器官损伤模型中维持心脏和肾脏功能，显著降低总体死亡率，强烈表明 24 在治疗与心肾综合征相关的心血管疾病中具有积极作用。（40）

Conclusion 结论

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In summary, we have identified 24 as a potent, orally available stimulator of sGC. Our optimization work starting from 1 led to the identification of compounds with superior in vitro and in vivo pharmacokinetic and metabolic profiles. 24, the compound with the best overall profile from these preclinical studies, was selected as clinical candidate and proved to have a pharmacokinetic profile in humans suitable for once daily dosing. Additional in vivo studies in animal models of hypertension, heart failure, and kidney disease have revealed dose-dependent antifibrotic and organ-protective properties in line with the sGC stimulator mode of action. 24 is currently being investigated in a phase 3 clinical trial in HFrEF patients.
总结来说，我们已确定 24 号化合物是一种强效、可口服的 sGC 激动剂。从 1 号化合物开始的优化工作，使我们鉴定出具有优越的体外和体内药代动力学及代谢特征的化合物。在这些临床前研究中，24 号化合物因其最佳的整体特征被选为临床候选药物，并在人体中表现出适合每日一次给药的药代动力学特征。在高血压、心力衰竭和肾脏疾病动物模型中的进一步体内研究揭示了 24 号化合物具有剂量依赖性的抗纤维化和器官保护作用，这与 sGC 激动剂的作用模式一致。目前，24 号化合物正在 HFrEF 患者中进行 3 期临床试验。

Experimental Section 实验部分

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Chemistry 化学

General Procedures 通用程序

Unless otherwise noted, all nonaqueous reactions were carried out under an argon atmosphere with commercial-grade reagents and solvents. All final products were at least 95% pure, as determined by analytical HPLC.
除非另有说明，所有非水溶液反应均在氩气气氛下，使用商业级试剂和溶剂进行。所有最终产品纯度至少为 95%，通过分析高效液相色谱法确定。

¹H NMR spectra were recorded on Bruker Avance spectrometers operating at 400 or 500 MHz. Chemical shifts (δ) are reported in ppm relative to TMS as an internal standard, and coupling constants (J) are given in hertz (Hz). Spin multiplicities are reported as s = singlet, br s = broad singlet, d = doublet, dd = doublet of doublets, t = triplet, q = quartet, m = multiplet. LC-MS analysis was performed using the respective method a–d, as noted. Method a: instrument, Micromass TOF-MUX interface 4× parallel injection with Waters 600 HPLC; column, Phenomenex Synergi 2 μm Hydro-RP Mercury 20 mm × 4 mm; mobile phase A, H₂O (1 L) + 50% formic acid (0.5 mL); mobile phase B, MeCN (1 L) + 50% formic acid (0.5 mL); gradient, 0.0 min 90% A → 2.5 min 30% A → 3.0 min 5%A → 4.5 min 5% A; oven, 20 °C; flow, 1 mL/min; UV detection, 210 nm. Method b: instrument, Micromass Quattro Premier with Waters Acquity UPLC; column, Thermo Hypersil GOLD 1.9 μm, 50 mm × 1 mm; mobile phase A, H₂O (1 L) + 50% formic acid (0.5 mL); mobile phase B, MeCN (1 L) + 50% formic acid (0.5 mL); gradient, 0.0 min 90% A → 0.1 min 90% A → 1.5 min 10% A → 2.2 min 10% A; oven, 50 °C; flow, 0.33 mL/min; UV detection, 210 nm. Method c: instrument, Waters Micromass Quattro Micro with Agilent 1100 series HPLC; column, Thermo Hypersil GOLD 3 μm, 20 mm × 4 mm; mobile phase A, H₂O (1 L) + 50% formic acid (0.5 mL); mobile phase B, MeCN (1 L) + 50% formic acid (0.5 mL); gradient, 0.0 min 100% A → 3.0 min 10% A → 4.0 min 10% A; ove,: 50 °C; flow, 2 mL/min; UV detection, 210 nm. Method d: instrument, Waters Acquity SQD UPLC system; column, Waters Acquity UPLC HSS T3 1.8 μm, 50 mm × 1 mm; mobile phase A, H₂O (1 L) + 99% formic acid (0.25 mL); mobile phase B, MeCN (1 L) + 99% formic acid (0.25 mL); gradient, 0.0 min 90% A → 1.2 min 5% A → 2.0 min 5% A; oven, 50 °C; flow, 0.4 mL/min; UV detection, 208–400 nm.

Materials 材料

Intermediates 1a–c, (25)3b, (33)4h, (41) and compounds 1, (25)2, (42)4–8, (43)9, (44)10, (44)11–14, (45)15–18, (43)19, (46)20, (47) and 23 (48) were synthesized according to the methods described previously.
中间体 1a-c、(25)3b、(33)4h、(41)以及化合物 1、(25)2、(42)4-8、(43)9、(44)10、(44)11-14、(45)15-18、(43)19、(46)20、(47)和 23（48）均按照先前描述的方法合成。

Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}(2-hydroxyethyl)carbamate (3)
甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}(2-羟基乙基)甲酰胺(3)

1c (200 mg, 0.57 mmol) and AcOH (33 μL, 0.57 mmol) were dissolved in MeOH (20 mL), and the mixture was cooled to 0 °C. Then a solution of glycolaldehyde (34 mg, 0.57 mmol) in MeOH (20 mL) was added dropwise. After complete addition, the mixture was stirred for a further 15 min at 0 °C. Then NaBH₃CN (50 mg, 0.80 mmol) was added portionwise. The mixture was stirred at rt for 20 h and then concentrated under reduced pressure. The residue was diluted with EtOAc and washed with concd aq NH₄Cl solution. The organic layer was concentrated under reduced pressure, and the residue was purified by HPLC (MeCN/H₂O gradient) to give 2-({4,6-diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}amino)ethanol (1d) as a yellow solid. Yield 165 mg (73%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.04 (dd, J = 8.1, 1.7 Hz, 1H), 8.58 (dd, J = 4.7, 1.7 Hz, 1H), 7.27–7.39 (m, 2H), 7.22 (dd, J = 10.0, 8.8 Hz, 1H), 7.08–7.16 (m, 2H), 6.09 (br s, 4H), 5.77 (s, 2H), 4.77 (t, J = 5.1 Hz, 1H), 3.47 (td, J = 5.6, 5.1 Hz, 2H), 3.27 (t, J = 6.4 Hz, 1H), 2.83 (dt, J = 6.4, 5.6 Hz, 2H). LC-MS (method a): t_R (min) = 1.27. MS (EI+): m/z = 395 [M + H]⁺.

1d (265 mg, 0.67 mmol) was dissolved in pyridine (5.0 mL), and the solution was cooled to 0 °C. Methyl chloroformate (83 mg, 0.87 mmol) was added, and the mixture was stirred at rt overnight. Then, the mixture was concentrated under reduced pressure, and the residue was purified by preparative HPLC (H₂O/MeCN/1% TFA 56:30:14) to give 3 as a white solid. Yield 37 mg (12%). ¹H NMR (500 MHz, DMSO-d₆): δ = 9.04 (dd, J = 8.0, 1.5 Hz, 1H), 8.59 (dd, J = 4.5, 1.5 Hz, 1H), 7.34–7.38 (m, 1H), 7.32 (dd, J = 8.0, 4.5 Hz, 1H), 7.22 (dd, J = 10.0, 8.8 Hz, 1H), 7.09–7.15 (m, 2H), 6.07 (br s, 4H), 5.78 (s, 2H), 4.14 (t, J = 5.7 Hz, 2H), 3.70 (s, 3H), 3.54 (t, J = 7.0 Hz, 1H), 3.00 (dt, J = 7.0, 5.7 Hz, 2H). LC-MS (method a): t_R (min) = 1.49. MS (EI+): m/z = 453 [M + H]⁺.

3-Bromo-1H-pyrazolo[3,4-c]pyridazine (3c)
3-溴-1H-吡唑并[3,4-c]吡啶杂氮（3c）

1H-Pyrazolo[3,4-c]pyridazin-3-ol (33) (3b; 18.00 g, 132.24 mmol) was dissolved in sulfolane (176 mL), and POBr₃ (39.81 g, 138.85 mmol) was then added to the solution. The mixture was stirred at 150 °C for 3 h. After being cooled to rt, the mixture was poured onto ice–water and extracted with EtOAc (3×). The combined organic extracts were washed with H₂O and brine, dried over Na₂SO₄, filtered, and concentrated until a precipitate formed. The solids were collected by filtration and dried under reduced pressure. Yield 16.27 g (62%). ¹H NMR (400 MHz, DMSO-d₆): δ = 14.90 (br s, 1H), 9.23 (d, J = 5.6 Hz, 1H), 8.10 (d, J = 5.6 Hz, 1H). LC-MS (method b): t_R (min) = 1.15. MS (ESI+): m/z = 199 [M + H]⁺.
1H-Pyrazolo[3,4-c]pyridazin-3-ol（33）（3b；18.00 克，132.24 摩尔）溶于磺化物（176 毫升），然后向溶液中加入 POBr3（39.81 克，138.85 摩尔）。混合物在 150°C 下搅拌 3 小时。冷却至室温后，将混合物倒入冰水中，用乙酸乙酯（EtOAc）萃取（3 次）。合并有机萃取物，用水和盐水洗涤，经无水硫酸钠干燥，过滤，浓缩至有沉淀形成。通过过滤收集固体，并在减压下干燥。产率 16.27 克（62%）。1H NMR（400 MHz，DMSO-d6）：δ = 14.90（br s，1H），9.23（d，J = 5.6 Hz，1H），8.10（d，J = 5.6 Hz，1H）。LC-MS（方法 b）：tR（分钟）= 1.15。MS（ESI+）：m/z = 199 [M + H]+。

3-Bromo-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine (3d)
3-溴-1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪（3d）

3c (16.27 g, 81.75 mmol) and Cs₂CO₃ (31.97 g, 98.10 mmol) were mixed in DMF (150 mL). A solution of 2-fluorobenzyl bromide (17.00 g, 89.93 mmol) in DMF (50 mL) was then added dropwise, and the mixture was stirred at rt overnight. The mixture was then diluted with EtOAc (500 mL) and washed with H₂O (3×) and brine (1×). The organic phase was separated, dried over MgSO₄, filtered, and concentrated. The residue was purified by flash column chromatography on silica gel (DCM/MeOH gradient). The obtained crude product was triturated with MTBE and DCM, collected by filtration, washed with MTBE, and dried under reduced pressure. Yield 11.36 g (45%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.30 (d, J = 5.6 Hz, 1H), 8.15 (d, J = 5.6 Hz, 1H), 7.35–7.44 (m, 2H), 7.16–7.28 (m, 2H), 5.96 (s, 2H). LC-MS (method c): t_R (min) = 1.11. MS (ESI+): m/z = 307 [M + H]⁺.
3c（16.27 克，81.75 毫摩尔）和 Cs2CO3（31.97 克，98.10 毫摩尔）在 150 毫升 DMF 中混合。随后，将 2-氟苄基溴（17.00 克，89.93 毫摩尔）的 DMF 溶液（50 毫升）逐滴加入，混合物在室温下搅拌过夜。然后，用 500 毫升乙酸乙酯稀释混合物，并用水（3 次）和盐水（1 次）洗涤。将有机相分离，用 MgSO4 干燥，过滤，浓缩。将残留物通过硅胶柱色谱（DCM/MeOH 梯度）纯化。得到的粗产品与 MTBE 和 DCM 研磨，过滤收集，用 MTBE 洗涤，减压干燥。产率 11.36 克（45%）。1H NMR（400 MHz，DMSO-d6）：δ = 9.30（d，J = 5.6 Hz，1H），8.15（d，J = 5.6 Hz，1H），7.35–7.44（m，2H），7.16–7.28（m，2H），5.96（s，2H）。LC-MS（方法 c）：tR（分钟）= 1.11。MS（ESI+）：m/z = 307 [M + H]+。

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carbonitrile (3e)
1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基腈（3e）

3d (1.00 g, 3.26 mmol) and CuCN (0.32 g, 3.58 mmol) were treated with anhyd DMSO (10 mL), and the mixture was stirred at 150 °C for 9 h. The mixture was brought to rt, then poured onto ice–water and treated with 25% aq NH₄OH until all solids were dissolved (blue solution). The mixture was then extracted with EtOAc (3×). The combined organic extracts were dried over Na₂SO₄, filtered, and concentrated. The residue was purified by flash column chromatography on silica gel (cyclohexane/EtOAc 4:1) to give the crude product which was used without further purification. Yield 0.57 g (51%, purity 74%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.45 (d, J = 5.9 Hz, 1H), 8.50 (d, J = 5.9 Hz, 1H), 7.39–7.51 (m, 2H), 7.18–7.30 (m, 2H), 6.11 (s, 2H). LC-MS (method d): t_R (min) = 0.94. MS (ESI+): m/z = 254 [M + H]⁺.
3d（1.00 克，3.26 毫摩尔）和 CuCN（0.32 克，3.58 毫摩尔）与无水 DMSO（10 毫升）混合，在 150°C 下搅拌 9 小时。将混合物冷却至室温，倒入冰水中，用 25%的氨水溶液处理至所有固体溶解（蓝色溶液）。然后将混合物用乙酸乙酯（3 次）萃取。将合并的有机层干燥后用无水硫酸钠过滤，浓缩。得到的残留物通过硅胶柱色谱（环己烷/乙酸乙酯 4:1）纯化，得到粗产品，未进一步纯化即使用。产率 0.57 克（51%，纯度 74%）。1H NMR（400 MHz，DMSO-d6）：δ = 9.45（d，J = 5.9 Hz，1H），8.50（d，J = 5.9 Hz，1H），7.39–7.51（m，2H），7.18–7.30（m，2H），6.11（s，2H）。LC-MS（方法 d）：tR（分钟）= 0.94。MS（ESI+）：m/z = 254 [M + H]+。

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carboximidamide (3f)
1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-羧基脒（3f）

3e (1.19 g, 4.70 mmol) and NaOMe (0.96 g, 17.86 mmol) were treated with anhyd MeOH (31 mL), and the mixture was stirred at rt for 2 h. NH₄Cl (0.30 g, 5.64 mmol) and AcOH (1.05 mL) were then added, and the mixture was stirred under reflux overnight. The mixture was then concentrated, and the residue was taken up in EtOAc and 1 M aq NaOH. The organic phase was separated, washed with 1 M aq NaOH, dried over Na₂SO₄, filtered, and concentrated. The solid residue was triturated with MTBE, collected by filtration, washed with DCM/MeOH (50:1), and dried under reduced pressure. Yellow solid; yield 0.43 g (34%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.26 (d, J = 5.1 Hz, 1H), 8.48 (d, J = 5.1 Hz, 1H), 7.38 (d, J = 5.9 Hz, 1H), 7.11–7.32 (m, 3H), 6.71 (br s, 2H), 5.99 (s, 2H). LC-MS (method c): t_R (min) = 0.52. MS (ESI+): m/z = 271 [M + H]⁺.

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (3i)
2-[(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪-3-基]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（3i）

Aniline (0.202 mL, 2.22 mmol) was dissolved in H₂O (2.0 mL), and the solution was cooled to 0 °C. Aqueous HCl (37%, 0.38 mL) was then added dropwise, followed by a solution of NaNO₂ (153 mg, 2.22 mmol) in H₂O (0.5 mL). The mixture was stirred for an additional 15 min, and a solution of NaOAc (231 mg, 2.82 mmol) in H₂O (0.5 mL) was then added dropwise followed by a solution of malononitrile (147 mg, 2.22 mmol) in EtOH (3.0 mL). The mixture was then stirred for an additional 2 h at 0 °C. The resulting precipitate was collected by filtration, washed with H₂O (3×), and then dissolved in DMF (5.0 mL). In a separate flask, 3f (599 mg, 2.22 mmol) was dissolved in DMF (5.0 mL), then Et₃N (0.309 mL, 2.22 mmol) was added dropwise. The mixture was warmed to 85 °C, and the aforementioned DMF solution was added. The resulting mixture was stirred at 100 °C overnight then cooled to 0 °C and treated with H₂O. The formed solids were collected by filtration, washed with H₂O and MeOH, and dried to give 2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine as a yellow solid. Yield 0.68 g (68%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.35 (d, J = 5.4 Hz, 1H), 9.01 (d, J = 5.6 Hz, 1H), 8.54 (br s, 2H), 7.89–8.07 (m, J = 7.8 Hz, 4H), 7.45–7.54 (m, 2H), 7.31–7.44 (m, 3H), 7.16–7.30 (m, 2H), 6.06 (s, 2H). LC-MS (method d): t_R (min) = 1.08. MS (ESI+): m/z = 441 [M + H]⁺.

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidine-4,5,6-triamine (3h)
2-[1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶并[3,4-c]三胺]-4,5,6-三胺（3h）

3i (300 mg, 0.68 mmol) was suspended in DMF (15 mL) and 10% Pd/C (58 mg) was added. The mixture was stirred in an H₂ atmosphere at ambient pressure overnight. The solids were collected by filtration and washed with MeOH and DCM, and the filtrate was concentrated under reduced pressure. The residue was triturated with DCM, and the solids were collected by filtration and dried under reduced pressure at 50 °C to give 2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidine-4,5,6-triamine. The crude product was used without further purification. Yield 198 mg (74%, purity 90%). LC-MS (method d): t_R (min) = 1.08. MS (ESI+): m/z = 352 [M + H]⁺.
3i（300 毫克，0.68 毫摩尔）悬浮于 15 毫升 DMF 中，加入 10% Pd/C（58 毫克）。混合物在常压下的 H2 气氛中搅拌过夜。通过过滤收集固体，并用甲醇和二氯甲烷洗涤，然后在减压下浓缩滤液。将残渣与二氯甲烷研磨，通过过滤收集固体，并在 50°C 下减压干燥，得到 2-[1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基]嘧啶-4,5,6-三胺。粗产品未经进一步纯化即使用。产率 198 毫克（74%，纯度 90%）。液相色谱-质谱（方法 d）：保留时间（分钟）= 1.08。质谱（ESI+）：相对分子质量 m/z = 352 [M + H]+。

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (4i)
2-[8-(2-氟苯基咪唑[1,5-a]嘧啶-6-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（4i）

8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidine-6-carboximidamide (41) (4h; 70% purity, 1.30 g, 3.40 mmol) was mixed with DMF (24.6 mL) and Et₃N (0.52 mL, 3.74 mmol). At 85 °C, a solution of [(E)-phenyldiazenyl]malononitrile (1b; 694 mg, 4.08 mmol) in DMF (12.0 mL) was added dropwise and the mixture was stirred for an additional 15 h at 100 °C. The resulting mixture was then poured into H₂O (300 mL), and the resulting precipitate was collected by filtration, washed with H₂O, and dried. Then, MeCN (160 mL) was added and the remaining solids were collected by filtration and dried again to give 2-[8-(2-fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine. Yield 419 mg (28%). ¹H NMR (400 MHz, DMSO-d₆): δ = 10.23–10.31 (m, 1H), 9.42 (br s, 1H), 8.60–8.66 (m, 1H), 8.12 (d, J = 7.6 Hz, 2H), 7.51–7.58 (m, 2H), 7.44–7.50 (m, 1H), 7.23–7.36 (m, 3H), 7.15–7.23 (m, 1H), 7.08–7.15 (m, 1H), 4.44 (s, 2H). LC-MS (method c): t_R (min) = 1.14. MS (ESI+): m/z = 440 [M + H]⁺.

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidine-4,5,6-triamine (4j)
2-[8-(2-氟苯基)咪唑并[1,5-a]嘧啶-6-基]嘧啶-4,5,6-三胺（4j）

4i (100 mg, 0.228 mmol) was dissolved in DMF (11.4 mL) and MeOH (2.9 mL). The solution was cooled with ice, and then 10% Pd/C (20 mg) was added. The mixture was stirred at 0 °C under 1 atm of H₂ for 48 h. The resulting mixture was filtered through Celite, and the solids were washed with MeOH. The combined filtrate was concentrated under reduced pressure to give crude 2-[8-(2-fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidine-4,5,6-triamine, which was purified by preparative reversed-phase HPLC (H₂O + 0.1% NH₃/MeCN gradient). Brown solid; yield 36 mg (45%). ¹H NMR (400 MHz, DMSO-d₆): δ = 10.13 (br d, J = 7.4 Hz, 1H), 8.20 (br d, J = 3.9 Hz, 1H), 7.19–7.31 (m, 2H), 7.04–7.17 (m, 2H), 6.81 (dd, J = 3.7, 7.6 Hz, 1H), 5.89 (s, 4H), 4.27 (s, 2H), 4.06 (s, 2H). LC-MS (method d): t_R (min) = 0.74. MS (ESI+): m/z = 351 [M + H]⁺.
4i（100 毫克，0.228 毫摩尔）溶解于 DMF（11.4 毫升）和 MeOH（2.9 毫升）中。溶液冷却至冰点，然后加入 10% Pd/C（20 毫克）。混合物在 0°C、1 个大气压的 H2 下搅拌 48 小时。所得混合物通过 Celite 过滤，固体用 MeOH 洗涤。合并滤液，减压浓缩，得到粗品 2-[8-(2-氟苯基)咪唑[1,5-a]嘧啶-6-基]嘧啶-4,5,6-三胺，经制备型反相高效液相色谱（H2O + 0.1% NH3/MeCN 梯度）纯化。棕色固体；产率 36 毫克（45%）。1H NMR（400 MHz，DMSO-d6）：δ = 10.13（br d，J = 7.4 Hz，1H），8.20（br d，J = 3.9 Hz，1H），7.19–7.31（m，2H），7.04–7.17（m，2H），6.81（dd，J = 3.7，7.6 Hz，1H），5.89（s，4H），4.27（s，2H），4.06（s，2H）。LC-MS（方法 d）：tR（分钟）= 0.74。MS（ESI+）：m/z = 351 [M + H]+。

4-(2,2,3,3-Tetrafluoropropyl)morpholine (6b)
4-(2,2,3,3-四氟丙基)吗啉（6b）

Tf₂O (252.5 g, 0.895 mol) was heated to 40 °C and, at this temperature, 2,2,3,3-tetrafluoro-1-propanol (6a; 130.0 g, 0.984 mol) was metered in while cooling. After the addition was completed, the mixture was heated to 70–75 °C and stirred for 2 h. Then the mixture was cooled to 20 °C, and the reaction solution of 2,2,3,3-tetrafluoropropyl trifluoromethanesulfonate was used without further purification.
Tf2O（252.5 克，0.895 摩尔）被加热至 40°C，在此温度下，2,2,3,3-四氟-1-丙醇（6a；130.0 克，0.984 摩尔）被计量加入，同时冷却。加入完成后，混合物被加热至 70-75°C，搅拌 2 小时。然后混合物被冷却至 20°C，未进一步纯化的 2,2,3,3-四氟丙基三氟甲烷磺酸反应溶液被使用。

Morpholine (158.5 g, 1.82 mol) was cooled to 5 °C. At 5–10 °C, the reaction solution of the triflate (189.5 g, max 0.455 mol) was added dropwise while cooling and then the mixture was stirred at 5–10 °C for 30 min. Then, the mixture was heated to 40 °C and stirred for 1 h. After cooling to 20 °C, H₂O (160 mL) and toluene (160 mL) were added and the phases were separated. The organic phase was washed with H₂O (160 mL) and concentrated on a rotary evaporator at 50 °C/50 mbar. The residue (81.0 g) was distilled at 67–68 °C/18 mbar to give 6b. Yield 77.0 g (84%). ¹H NMR (400 MHz, CDCl₃): δ = 5.83–6.22 (m, 1H), 3.61–3.78 (m, 4H), 2.89 (tt, J = 14.0, 1.7 Hz, 2H), 2.53–2.70 (m, 4H).
Morpholine（158.5 克，1.82 摩尔）被冷却至 5°C。在 5-10°C 时，将三氟甲磺酸酯（189.5 克，最大 0.455 摩尔）逐滴加入并继续冷却，然后在 5-10°C 下搅拌 30 分钟。之后，将混合物加热至 40°C，搅拌 1 小时。冷却至 20°C 后，加入 160 毫升水和 160 毫升甲苯，分离两相。用 160 毫升水洗涤有机相，并在 50°C/50 mbar 下用旋转蒸发器浓缩。得到的残留物（81.0 克）在 67-68°C/18 mbar 下蒸馏，得到 6b。产率 77.0 克（84%）。1H NMR（400 MHz，CDCl3）：δ = 5.83-6.22（m，1H），3.61-3.78（m，4H），2.89（tt，J = 14.0，1.7 Hz，2H），2.53-2.70（m，4H）。

4-Methyl-4-(2,2,3,3-tetrafluoropropyl)morpholin-4-ium Methanesulfonate (6c)
4-甲基-4-(2,2,3,3-四氟丙基)吗啉-4-ium 甲磺酸盐（6c）

Methylmethanesulfonate (143.7 g, 1.31 mol) was heated to 135 °C and, at this temperature, 6b (250.0 g, 1.24 mol) was added dropwise. The mixture was stirred at 100 °C for 22 h, then cooled to 85 °C and i-PrOH (375 mL) was added. After cooling to 0–5 °C, the mixture was stirred for a further 30 min. The product was collected by suction filtration, washed with i-PrOH (3 × 125 mL), and dried at 45 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 336.8 g (87%). ¹H NMR (400 MHz, D₂O): δ = 6.13–6.48 (m, 1H), 4.33–4.51 (m, 2H), 4.01–4.24 (m, 4H), 3.68–3.93 (m, 4H), 3.55 (s, 3H), 2.81 (s, 3H).
甲基甲烷磺酸（143.7 克，1.31 摩尔）加热至 135°C，在此温度下，6b（250.0 克，1.24 摩尔）逐滴加入。混合物在 100°C 下搅拌 22 小时，然后冷却至 85°C，加入 375 毫升-PrOH。冷却至 0-5°C 后，继续搅拌 30 分钟。产品通过吸滤收集，用-PrOH（3×125 毫升）洗涤，并在 45°C 下，在真空干燥柜中用温和的 N2 气流干燥。产率 336.8 克（87%）。1H NMR（400 MHz，D2O）：δ = 6.13–6.48（m，1H），4.33–4.51（m，2H），4.01–4.24（m，4H），3.68–3.93（m，4H），3.55（s，3H），2.81（s，3H）。

4-Methyl-4-(2,3,3-trifluoroprop-1-enyl)morpholin-4-ium Methanesulfonate (6d)
4-甲基-4-(2,3,3-三氟丙-1-烯基)吗啉-4-ium 甲磺酸盐（6d）

First, 45% aq NaOH (16.9 g, 189.9 mmol) was metered into a solution of 6c (53.8 g, 172.7 mol) in H₂O (40 mL) at 50–55 °C, and the mixture was stirred at 50 °C for 1 h then cooled to 20 °C. Then the precipitated salts were removed by suction filtration and washed with H₂O (5 mL). The aqueous solution of product 6d (102.1 g, max 172.7 mmol) was used in the next stage. For analytical purposes, a sample was concentrated and dried. ¹H NMR (400 MHz, D₂O): δ = 6.74–6.83 (m, 1H), 6.39–6.69 (m, 1H), 4.12–4.20 (m, 2H), 3.97–4.09 (m, 4H), 3.76–3.85 (m, 2H), 3.59 (s, 3H), 2.81 (s, 3H).
首先，将 45%的 NaOH 溶液（16.9 克，189.9 毫摩尔）加入 6c 溶液（53.8 克，172.7 摩尔）中，该溶液在 40 毫升水中，温度控制在 50-55°C，混合物在 50°C 下搅拌 1 小时，然后冷却至 20°C。随后，通过吸滤去除沉淀的盐，并用 5 毫升水洗涤。产品 6d 的溶液（102.1 克，最大 172.7 毫摩尔）用于下一阶段。为了分析目的，取样品浓缩并干燥。1H NMR（400 MHz，D2O）：δ = 6.74-6.83（m，1H），6.39-6.69（m，1H），4.12-4.20（m，2H），3.97-4.09（m，4H），3.76-3.85（m，2H），3.59（s，3H），2.81（s，3H）。

2-Fluoro-3-(morpholin-4-yl)acrylaldehyde (6e)
2-氟-3-(4-吗啉基)丙烯醛（6e）

A mixture of morpholine (30.2 g, 345.3 mmol) and Et₃N (52.5 g, 518.0 mmol) was heated to 75 °C, and the aqueous solution of 6d (max 172.7 mmol) was added dropwise at 75–80 °C. The mixture was stirred under reflux for 2 h, cooled to 23 °C, and washed with DCM (100 mL). The aqueous phase was washed twice with DCM/Et₃N (100:15, 115 mL), and the combined organic phases were washed with satd aq K₂CO₃ solution (85 mL) and concentrated under reduced pressure at 45–50 °C. Toluene (120 mL) was added, then toluene (60 mL) was distilled off. The suspension was stirred at rt overnight, and the product was collected by suction filtration and dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 19.2 g (68%). ¹H NMR (500 MHz, CDCl₃): δ = 8.59 (d, J = 18.9 Hz, 1H), 6.16 (d, J = 27.1 Hz, 1H), 3.72–3.83 (m, 4H), 3.51–3.60 (m, 4H).
一种由 30.2 克（345.3 毫摩尔）的哌啶和 52.5 克（518.0 毫摩尔）的 Et3N 混合物在 75°C 下加热，然后逐滴加入最多 172.7 毫摩尔的 6d 水溶液，温度保持在 75-80°C。混合物在回流条件下搅拌 2 小时，冷却至 23°C，并用 100 毫升二氯甲烷（DCM）洗涤。水相用二氯甲烷/三乙胺（100:15，115 毫升）洗涤两次，合并后的有机相用饱和碳酸钾溶液（85 毫升）洗涤，并在 45-50°C 下减压浓缩。加入 120 毫升甲苯，然后蒸馏掉 60 毫升甲苯。将悬浮液在室温下搅拌过夜，通过吸滤收集产物，并在 50°C 下用氮气流在真空干燥柜中干燥。产率 19.2 克（68%）。1H NMR（500 MHz，CDCl3）：δ = 8.59（d，J = 18.9 Hz，1H），6.16（d，J = 27.1 Hz，1H），3.72-3.83（m，4H），3.51-3.60（m，4H）。

Ethyl 5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxylate (6f)
乙基 5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酸酯（6f）

Ethyl 5-amino-1-(2-fluorobenzyl)-1H-pyrazole-3-carboxylate (49) (22.3 g, 84.8 mmol) was initially charged into EtOH (59.5 mL), and MsOH (11.0 mL, 169.6 mmol), LiCl (9.0 g, 212.1 mmol), and 6e (15.0 g, 84.8 mmol) were added at rt. The mixture was stirred under reflux for 4.5 h. After cooling to rt, the product was collected by suction filtration, washed with EtOH (2 × 4.5 mL), and stirred with H₂O (325 mL) for 1 h. The solids were collected by suction filtration, washed with H₂O (2 × 11.5 mL), and dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 21.8 g (81%). ¹H NMR (400 MHz, CDCl₃): δ = 8.51 (dd, J = 2.7, 1.7 Hz, 1H), 8.15 (dd, J = 7.7, 2.8 Hz, 1H), 7.19–7.33 (m, 1H), 6.93–7.16 (m, 3H), 5.88 (s, 2H), 4.52 (q, J = 7.2 Hz, 2H), 1.48 (t, J = 7.2 Hz, 3H). MS (ESI+): m/z = 318 [M + H]⁺.

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxamide (6g)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酰胺（6g）

EtOH (10 mL), formamide (14.9 mL, 441.2 mmol), and 30% NaOMe in MeOH (3.6 g, 19.8 mmol) were added to 6f (7.0 g, 22.1 mmol). The reaction mixture was heated to 95–100 °C, and the volatile solvents were distilled off by a downward distillation within 1 h. The mixture was stirred at 125 °C for 1.5 h, H₂O (30 mL) was added, and the resulting mixture was cooled to rt and stirred for 1 h. The precipitated solids were collected by suction filtration, washed with H₂O (3 × 8.5 mL), and dried at 45 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 6.2 g (97%). ¹H NMR (400 MHz, DMSO-d₆): δ = 8.72 (dd, J = 2.7, 1.7 Hz, 1H), 8.28 (dd, J = 8.3, 2.8 Hz, 1H), 7.87 (br s, 1H), 7.60 (br s, 1H), 7.34–7.40 (m, 1H), 7.12–7.26 (m, 3H), 5.87 (s, 2H). MS (ESI+): m/z = 289 [M + H]⁺.
乙醇（10 mL）、甲酰胺（14.9 mL，441.2 mmol）和 30% NaOMe 的甲醇溶液（3.6 g，19.8 mmol）加入 6f（7.0 g，22.1 mmol）。反应混合物加热至 95-100°C，在 1 小时内通过向下蒸馏去除挥发性溶剂。混合物在 125°C 下搅拌 1.5 小时，加入 30 mL 水，然后将混合物冷却至室温并搅拌 1 小时。通过吸滤收集析出的固体，用 8.5 mL 水洗涤 3 次，并在真空干燥柜中在 45°C 下用温和的氮气流干燥。产率 6.2 g（97%）。1H NMR（400 MHz，DMSO-d6）：δ = 8.72（dd，J = 2.7，1.7 Hz，1H），8.28（dd，J = 8.3，2.8 Hz，1H），7.87（br s，1H），7.60（br s，1H），7.34-7.40（m，1H），7.12-7.26（m，3H），5.87（s，2H）。MS（ESI+）：m/z = 289 [M + H]+。

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile (6h)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基腈（6h）

6g (17.3 g, 60.0 mmol) was heated to 103–107 °C in sulfolane (40.5 mL) and MeCN (5.4 mL). POCl₃ (6.9 g, 45.0 mmol) was slowly added dropwise while stirring, the dropping funnel was rinsed with MeCN (2.8 mL), and then the mixture was stirred at 107 °C for 1.5 h until conversion was complete (HPLC). Then the mixture was cooled to rt and sulfolane/MeCN (5:1, 2.8 mL) and then H₂O (17.8 mL) were added dropwise. The mixture was stirred for 0.5 h, a solution of aq NH₃ (28%, 9.4 g) in H₂O (22.7 mL) was added dropwise, and the resulting mixture was stirred for a further 2 h. The precipitated solids were collected by suction filtration, washed with H₂O (3 × 20.5 mL), and dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 14.7 g (92%). ¹H NMR (400 MHz, DMSO-d₆): δ = 8.87 (dd, J = 2.6, 1.7 Hz, 1H), 8.52 (dd, J = 8.1, 2.6 Hz, 1H), 7.17–7.42 (m, 4H), 5.87 (s, 2H). MS (ESI+): m/z = 271 [M + H]⁺.
6g（17.3g，60.0mmol）在 103～107°C 下用磺酰烷（40.5mL）和甲腈（5.4mL）加热。在搅拌下缓慢滴加 POCl3（6.9g，45.0mmol），滴液漏斗用甲腈（2.8mL）冲洗，然后在 107°C 下搅拌 1.5 小时，直至转化完全（HPLC）。然后冷却至室温，加入磺酰烷/甲腈（5:1，2.8mL）和 H2O（17.8mL），滴加。搅拌 0.5 小时，滴加 28%的氨水溶液（9.4g，22.7mL），继续搅拌 2 小时。通过吸滤收集沉淀固体，用 H2O（3×20.5mL）洗涤，在 50°C 下在真空干燥柜中用温和的 N2 气流干燥。产率 14.7g（92%）。1H NMR（400MHz，DMSO-d6）：δ=8.87（dd，J=2.6，1.7Hz，1H），8.52（dd，J=8.1，2.6Hz，1H），7.17–7.42（m，4H），5.87（s，2H）。MS（ESI+）：m/z=271 [M + H]+。

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboximidamide Hydrochloride (6i)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-羧基脒盐酸盐（6i）

6h (406.0 g, 1.50 mol) was suspended in EtOH (2.08 L). Then, 30% NaOMe in MeOH (54.1 g, 0.30 mol) was added and the mixture was stirred at rt overnight. NH₄Cl (88.4 g, 1.65 mol) was added, and the mixture was heated to 65 °C and stirred at 65 °C for 3.5 h. The solvents were distilled off, and the residue was stirred with EtOAc (1.60 L) overnight. The precipitated solids were collected by suction filtration, washed with EtOAc (2 × 140 mL), and dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 441.4 g (91%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.35 (br s, 3H), 8.86 (dd, J = 2.5, 1.5 Hz, 1H), 8.48 (dd, J = 8.8, 2.6 Hz, 1H), 7.36–7.43 (m, 1H), 7.29–7.35 (m, 1H), 7.22–7.28 (m, 1H), 7.15–7.20 (m, 1H), 5.90 (s, 2H). MS (ESI+): m/z = 288 [M + H]⁺.
6 小时（406.0 克，1.50 摩尔）的原料在乙醇（2.08 升）中悬浮。然后，加入 30%的 NaOMe 甲醇溶液（54.1 克，0.30 摩尔），混合物在室温下搅拌过夜。加入氯化铵（88.4 克，1.65 摩尔），将混合物加热至 65°C，并在 65°C 下搅拌 3.5 小时。蒸馏掉溶剂，将残留物与乙酸乙酯（1.60 升）搅拌过夜。通过抽滤收集析出的固体，用乙酸乙酯（2×140 毫升）洗涤，并在 50°C 下，在真空干燥柜中用温和的氮气流干燥。产率 441.4 克（91%）。1H NMR（400 MHz，DMSO-d6）：δ = 9.35（br s，3H），8.86（dd，J = 2.5，1.5 Hz，1H），8.48（dd，J = 8.8，2.6 Hz，1H），7.36–7.43（m，1H），7.29–7.35（m，1H），7.22–7.28（m，1H），7.15–7.20（m，1H），5.90（s，2H）。MS（ESI+）：m/z = 288 [M + H]+。

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (6j)
2-[(5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（6j）

Concd HCl (262 g, 2.59 mol) and H₂O (117.5 mL) were added dropwise at 0–5 °C to H₂O (1.525 L) and aniline (117.5 g, 1.26 mol). Then, a solution of NaNO₂ (87.1 g, 1.26 mol) in H₂O (222.5 mL) was added dropwise within 1 h, the dropping funnel was rinsed with H₂O (60 mL), and the mixture was stirred at 0–5 °C for 15 min. Thereafter, at this temperature, a solution of NaOAc (131.4 g, 1.60 mol) in H₂O (665 mL) was added dropwise within 45 min, the dropping funnel was rinsed with H₂O (60 mL), and a solution of malononitrile (83.4 g, 1.26 mol) in EtOH (233 mL) was added dropwise within 1 h. The dropping funnel was rinsed with EtOH (68.5 mL), and the mixture was further stirred at 0–5 °C for 2 h. The yellow solids were collected by suction filtration and washed with H₂O (3 × 625 mL) and cold toluene (488 mL). The still-moist residue was dissolved in DMF (872 g), which gave a DMF solution of [(E)-phenyldiazenyl]malononitrile (1.117 kg). 6i (30.0 g, 92.7 mmol) was suspended in DMF (72 mL). The mixture was heated to 100 °C, and a mixture of Et₃N (14.2 mL, 101.9 mmol) and the DMF solution of [(E)-phenyldiazenyl]malononitrile (150 g) was added dropwise at this temperature within 30 min. The dropping funnel was rinsed with DMF (30 mL), and the mixture was further stirred at 100 °C for 20 h. Then, it was cooled to 95–90 °C, H₂O (24 mL) was added dropwise within 10 min, and the resulting mixture was cooled to 0–5 °C within 1.5 h and stirred for 1 h. The solids were collected by suction filtration, washed with H₂O (60 mL)/DMF (63 mL), twice with H₂O (50 mL)/MeOH (63 mL), and then with MeOH (63 mL), suction-dried, and then dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 35.5 g (84%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.03 (dd, J = 8.8, 2.8 Hz, 1H), 8.65–8.77 (m, 1H), 8.50 (br s, 2H), 8.02 (d, J = 7.6 Hz, 2H), 7.86–7.98 (m, 2H), 7.44–7.57 (m, 2H), 7.32–7.44 (m, 2H), 7.11–7.31 (m, 3H), 5.84 (s, 2H). LC-MS (method d): t_R (min) = 1.15. MS (ESI+): m/z = 458 [M + H]⁺.
浓盐酸（262 克，2.59 摩尔）和水（117.5 毫升）在 0-5°C 下逐滴加入 1.525 升水中和苯胺（117.5 克，1.26 摩尔）。然后，在 1 小时内逐滴加入 87.1 克亚硝酸钠（1.26 摩尔）的水溶液（222.5 毫升），滴液漏斗用 60 毫升水冲洗，混合物在 0-5°C 下搅拌 15 分钟。之后，在此温度下，在 45 分钟内逐滴加入 131.4 克醋酸钠（1.60 摩尔）的水溶液（665 毫升），滴液漏斗用 60 毫升水冲洗，并逐滴加入 83.4 克丙二腈（1.26 摩尔）的乙醇溶液（233 毫升），在 1 小时内完成。滴液漏斗用 68.5 毫升乙醇冲洗，混合物在 0-5°C 下继续搅拌 2 小时。黄色固体通过吸滤收集，用 3×625 毫升水和 488 毫升冷甲苯洗涤。仍然潮湿的残留物溶解在 872 克二甲基甲酰胺中，得到[(E)-苯基叠氮基]丙二腈的二甲基甲酰胺溶液（1.117 千克）。6i（30.0 克，92.7 毫摩尔）悬浮在 72 毫升二甲基甲酰胺中。将混合物加热至 100°C，并在 30 分钟内逐滴加入 14.2 毫升三乙胺（101.9 毫摩尔）和[(E)-苯基叠氮基]丙二腈的二甲基甲酰胺溶液（150 克）。滴定漏斗用 DMF（30 mL）冲洗，混合物在 100°C 下搅拌 20 小时。然后，将其冷却至 95-90°C，在 10 分钟内逐滴加入 24 mL 水，所得混合物在 1.5 小时内冷却至 0-5°C，并搅拌 1 小时。通过吸滤收集固体，用 H2O（60 mL）/DMF（63 mL）洗涤，然后用 H2O（50 mL）/甲醇（63 mL）洗涤两次，接着用甲醇（63 mL）洗涤，吸滤干燥，然后在 50°C 下，在真空干燥柜中用温和的氮气流干燥。产率 35.5 g（84%）。1H NMR（400 MHz，DMSO-d6）：δ = 9.03（dd，J = 8.8，2.8 Hz，1H），8.65-8.77（m，1H），8.50（br s，2H），8.02（d，J = 7.6 Hz，2H），7.86-7.98（m，2H），7.44-7.57（m，2H），7.32-7.44（m，2H），7.11-7.31（m，3H），5.84（s，2H）。LC-MS（方法 d）：tR（min）= 1.15。MS（ESI+）：m/z = 458 [M + H]+。

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidine-4,5,6-triamine (6k)
2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-4,5,6-三胺（6k）

6j (182.0 g, 0.39 mol) was initially charged into DMF (1.82 L), and then 5% Pd/C (50% water-moist, 4.2 g) was added. Hydrogenation was effected at 60 °C and H₂ pressure of 60 bar while stirring overnight. The mixture was filtered through kieselguhr, and the solids were washed thoroughly with DMF (150 mL) and then with MeOH (150 mL). The filtrate was concentrated at 60–70 °C to a weight of 425 g of distillation residue. The residue was heated to 75–80 °C, MeOH (300 mL) was added dropwise at this temperature, and the mixture was stirred for 15 min. The mixture was cooled to rt within 1 h, then H₂O (1.29 L) was added dropwise and the mixture was stirred overnight. The solids were collected by suction filtration, washed with H₂O (2 × 500 mL), suction-dried, and then dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 159.7 g. For analytical purposes, a sample was purified by chromatography on silica gel (DCM/MeOH 9:1). ¹H NMR (400 MHz, DMSO-d₆): δ = 8.85 (dd, J = 9.0, 2.9 Hz, 1H), 8.62 (dd, J = 2.8, 1.7 Hz, 1H), 7.32–7.39 (m, 1H), 7.10–7.26 (m, 3H), 5.86 (br s, 4H), 5.75 (s, 2H), 4.04 (br s, 2H). MS (ESI+): m/z = 369 [M + H]⁺.
6j（182.0 克，0.39 摩尔）最初加入 1.82 升 DMF 中，然后加入 5% Pd/C（50%水湿，4.2 克）。在 60°C 和 60 bar 的 H2 压力下搅拌过夜进行加氢。混合物通过硅藻土过滤，固体用 DMF（150 毫升）和甲醇（150 毫升）彻底洗涤。滤液在 60-70°C 浓缩至 425 克蒸馏残渣。残渣加热至 75-80°C，在此温度下逐滴加入 300 毫升甲醇，混合物搅拌 15 分钟。混合物在 1 小时内冷却至室温，然后逐滴加入 1.29 升水，混合物搅拌过夜。固体通过吸滤收集，用水（2×500 毫升）洗涤，吸滤干燥，然后在真空干燥柜中在 50°C 下用温和的 N2 气流干燥。产率 159.7 克。为分析目的，样品通过硅胶柱层析（DCM/MeOH 9:1）进行纯化。1H NMR（400 MHz，DMSO-d6）：δ = 8.85（dd，J = 9.0，2.9 Hz，1H），8.62（dd，J = 2.8，1.7 Hz，1H），7.32-7.39（m，1H），7.10-7.26（m，3H），5.86（br s，4H），5.75（s，2H），4.04（br s，2H）。MS（ESI+）：m/z = 369 [M + H]+。

Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidin-5-yl}carbamate Formic Acid Salt (21)
甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基]嘧啶-5-基}甲酰胺甲酸盐（21）

The crude intermediate (3h: 198 mg, 0.51 mmol) was treated with pyridine (15.0 mL), and the mixture was cooled to 0 °C. Next, a solution of methyl chloroformate (43.6 μL, 0.57 mmol) in DCM (1.0 mL) was slowly added and the mixture was stirred at rt for 3 d. Additional methyl chloroformate (0.1 equiv) in DCM (1.0 mL) was added at 0 °C, and the mixture was stirred for a further 30 min at rt. The reaction mixture was then concentrated, and the residue was treated with MeCN. The resulting suspension was filtered, and the filtrate was concentrated. The residue was purified by preparative reversed-phase HPLC (H₂O + 0.05% formic acid/MeOH gradient) to give 21 as a white solid. Yield 72 mg (33%, purity 97%). ¹H NMR (400 MHz, DMSO-d₆): δ = 9.28 (d, J = 5.4 Hz, 1H), 8.88 (d, J = 5.6 Hz, 1H), 8.15 (s, 1H), 7.95–8.06 (m, 1H), 7.34–7.43 (m, 1H), 7.12–7.33 (m, 4H), 6.17–6.32 (m, 4H), 6.01 (s, 2H), 3.62 (s, 3H). LC-MS (method d): t_R (min) = 0.66. MS (ESI+): m/z = 410 [M + H]⁺.
粗中间体（3h：198 mg，0.51 mmol）与吡啶（15.0 mL）反应，混合物冷却至 0°C。随后，将甲基氯甲酸酯（43.6 μL，0.57 mmol）的 DCM（1.0 mL）溶液缓慢加入，混合物在室温下搅拌 3 天。在 0°C 下，再向混合物中加入 0.1 当量的甲基氯甲酸酯（DCM 1.0 mL），并在室温下继续搅拌 30 分钟。然后将反应混合物浓缩，将残留物用 MeCN 处理。得到的悬浮液经过滤，滤液浓缩。残留物通过制备型反相高效液相色谱（H2O + 0.05%甲酸/MeOH 梯度）纯化，得到 21 号化合物，为白色固体。产率 72 mg（产率 33%，纯度 97%）。1H NMR（400 MHz，DMSO-d6）：δ = 9.28（d，J = 5.4 Hz，1H），8.88（d，J = 5.6 Hz，1H），8.15（s，1H），7.95–8.06（m，1H），7.34–7.43（m，1H），7.12–7.33（m，4H），6.17–6.32（m，4H），6.01（s，2H），3.62（s，3H）。LC-MS（方法 d）：tR（min）= 0.66。MS（ESI+）：m/z = 410 [M + H]+。

Methyl {4,6-Diamino-2-[8-(2-fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidin-5-yl}carbamate (22)
甲基{4,6-二氨基-2-[8-(2-氟苯基)咪唑[1,5-a]嘧啶-6-基]嘧啶-5-基}甲酰胺（22）

4j (45 mg, 0.128 mmol) was dissolved in pyridine (3.46 mL), and the solution was cooled to 0 °C. Then, methyl chloroformate (12.9 μL, 0.167 mmol) was added and the mixture was stirred for an additional 5 min at 0 °C and for 19 h at rt. The resulting mixture was concentrated under reduced pressure to a volume of 1.5 mL, and the residue was purified by preparative reversed-phase HPLC (H₂O + 0.1% NH₃/MeCN gradient) to give 22 as a yellow solid. Yield 24 mg (45%). ¹H NMR (400 MHz, DMSO-d₆): δ = 10.22 (d, J = 7.15 Hz, 1H), 8.28 (d, J = 3.8 Hz, 1H), 7.97 (br s, 0.65 H), 7.67 (br s, 0.35 H), 7.20–7.29 (m, 2H), 7.11–7.18 (m, 1H), 7.05–7.11 (m, 1H), 6.89 (dd, J = 3.7, 7.3 Hz, 1H), 6.24 (br s, 4H), 4.30 (s, 2H), 3.61 (br s, 3H). LC-MS (method d): t_R (min) = 0.73. MS (ESI+): m/z = 409 [M + H]⁺.
4j（45 mg，0.128 mmol）溶解于吡啶（3.46 mL）中，溶液冷却至 0°C。然后加入甲基氯甲酸酯（12.9 μL，0.167 mmol），在 0°C 下搅拌 5 分钟，并在室温下搅拌 19 小时。将得到的混合物在减压下浓缩至 1.5 mL，并通过制备型反相高效液相色谱（H2O + 0.1% NH3/MeCN 梯度）纯化，得到 22 号化合物，为黄色固体。产率 24 mg（45%）。1H NMR（400 MHz，DMSO-d6）：δ = 10.22（d，J = 7.15 Hz，1H），8.28（d，J = 3.8 Hz，1H），7.97（br s，0.65 H），7.67（br s，0.35 H），7.20–7.29（m，2H），7.11–7.18（m，1H），7.05–7.11（m，1H），6.89（dd，J = 3.7，7.3 Hz，1H），6.24（br s，4H），4.30（s，2H），3.61（br s，3H）。LC-MS（方法 d）：tR（min）= 0.73。MS（ESI+）：m/z = 409 [M + H]+。

Methyl {4,6-Diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}carbamate (24)
甲基{4,6-二氨基-2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}甲酰胺（24）

6k (77% by weight, 4.0 g, 8.36 mmol) in i-PrOH (37.9 mL) was heated to 35 °C, and then methyl chloroformate (0.84 mL, 10.87 mmol) was added dropwise. The mixture was stirred at 35–40 °C for 20 h, heated to 50 °C, and MeOH (9.5 mL) was added. Then, Et₃N (1.9 mL) was added dropwise within 0.5 h, the dropping funnel was rinsed with MeOH (1.3 mL), and the resulting mixture was stirred at 50 °C for 1 h. Thereafter, the mixture was cooled to rt and stirred at rt for 1 h. The solids were collected by suction filtration, washed with EtOH (3 × 8 mL), suction-dried, and then dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet. Yield 3.4 g of crude product. The crude product (3.0 g) was stirred in DMSO (8 mL) for 5 min, EtOAc (13.0 mL) and activated carbon (50 mg) were added, and the mixture was heated under reflux (84 °C) for 15 min. The suspension was hot-filtered, and the filter residue was washed with EtOAc (1.9 mL). EtOAc (60 mL) and EtOH (16 mL) were heated to 60 °C, the combined filtrates were added dropwise, and the resulting mixture was stirred at 60 °C for 1.5 h. The suspension was cooled to rt within 25 min, stirred for a further 1.5 h, cooled further to 0–5 °C, and stirred for a further 1 h. The solids were collected by suction filtration, washed with EtOAc (2 × 6.4 mL), suction-dried, and then dried at 50 °C under a gentle N₂ stream in a vacuum drying cabinet to give 24. Yield 2.2 g (70%). ¹H NMR (400 MHz, DMSO-d₆): δ = 8.89 (dd, J = 9.0, 2.8 Hz, 1H), 8.66 (m, 1H), 7.99 and 7.67 (2 br s, 1H), 7.32–7.40 (m, 1H), 7.19–7.26 (m, 1H), 7.10–7.19 (m, 2H), 6.22 (br s, 4H), 5.79 (s, 2H), 3.62 (br s, 3H). LC-MS (method d): t_R (min) = 0.79. MS (ESI+): m/z = 427 [M + H]⁺.
6k（质量分数 77%，4.0 克，8.36 毫摩尔）在-PrOH（37.9 毫升）中加热至 35°C，然后逐滴加入甲基氯甲酸酯（0.84 毫升，10.87 毫摩尔）。混合物在 35-40°C 下搅拌 20 小时，加热至 50°C，加入甲醇（9.5 毫升）。接着，在 0.5 小时内逐滴加入三乙胺（1.9 毫升），用甲醇（1.3 毫升）冲洗分液漏斗，所得混合物在 50°C 下搅拌 1 小时。之后，将混合物冷却至室温，在室温下搅拌 1 小时。通过吸滤收集固体，用乙醇（3×8 毫升）洗涤，吸滤干燥，然后在真空干燥柜中在 50°C 下用温和的氮气流干燥。产率 3.4 克粗产品。将粗产品（3.0 克）在 DMSO（8 毫升）中搅拌 5 分钟，加入乙酸乙酯（13.0 毫升）和活性炭（50 毫克），混合物在回流（84°C）下加热 15 分钟。热过滤悬浮液，用乙酸乙酯（1.9 毫升）洗涤滤渣。将乙酸乙酯（60 毫升）和乙醇（16 毫升）加热至 60°C，将混合滤液逐滴加入，所得混合物在 60°C 下搅拌 1.5 小时。将悬浮液在 25 分钟内冷却至室温，再搅拌 1 小时。5 小时，进一步冷却至 0-5°C，并搅拌 1 小时。通过吸滤收集固体，用乙醚（2×6.4 mL）洗涤，吸滤干燥，然后在 50°C 下，在真空干燥柜中用温和的氮气流干燥，得到 24.产率 2.2 g（70%）。1H NMR（400 MHz，DMSO-d6）：δ = 8.89（dd，J = 9.0，2.8 Hz，1H），8.66（m，1H），7.99 和 7.67（2 br s，1H），7.32-7.40（m，1H），7.19-7.26（m，1H），7.10-7.19（m，2H），6.22（br s，4H），5.79（s，2H），3.62（br s，3H）。LC-MS（方法 d）：tR（min）= 0.79。MS（ESI+）：m/z = 427 [M + H]+。

Biology 生物

General 通用

Animal experiments were conducted in accordance with the German animal welfare laws, approved by local authorities, and in accordance with the ethical guidelines of Bayer AG.
动物实验是在遵守德国动物福利法、经当地当局批准以及符合拜耳公司伦理指南的情况下进行的。

CYP Inhibition Assay CYP 抑制测定法

The inhibitory potency of 24 was assessed in vitro by means on formation of metabolites from standard probes mediated by CYP isoforms (for details, please refer to the Supporting Information) based on assay conditions described. (50) To investigate time-dependency, preincubation experiments on CYP3A4 were performed. (51)
24 的抑制活性通过体外实验评估，该实验基于标准探针在 CYP 同工酶介导下形成代谢物的条件进行（具体细节请参阅补充材料）。为了研究时间依赖性，对 CYP3A4 进行了预孵育实验。（50）为了研究时间依赖性，对 CYP3A4 进行了预孵育实验。（51）

In Vitro Clearance Determinations with Rat and Human Hepatocytes
体外清除度测定：大鼠和人肝细胞

Incubations with hepatocytes were performed at 37 °C, pH 7.4, in a total volume of 1.5 mL using a modified Janus robotic system (PerkinElmer). The incubation mixtures contained 1 × 10⁶ cells/mL (corrected, according to the viability of the cells, determined via microscopy after staining with trypan blue), 1 μM substrate, and Williams’ medium E (Sigma, product no. W1878). The final MeCN concentration was ≤1%. Aliquots of 125 μL were withdrawn from the incubation mixture after 2, 10, 20, 30, 50, 70, and 90 min and dispensed in a 96-well plate, containing MeCN (250 μL) to stop the reaction. After centrifugation at 1000g, supernatants were analyzed by LC-MS/MS (AB Sciex Triple Quad 5500).
在 37°C、pH 7.4 的条件下，使用改良的 Janus 机器人系统（PerkinElmer）进行肝细胞的培养。培养混合物中含有 1×10^6 个细胞/mL（根据细胞活力校正，通过显微镜染色后用台盼蓝染色确定），1 μM 底物和 Williams’ E 培养基（Sigma，产品编号 W1878）。最终 MeCN 浓度≤1%。在 2、10、20、30、50、70 和 90 分钟后，从培养混合物中取出 125 μL 的样品，并将其分装到含有 MeCN（250 μL）的 96 孔板中，以终止反应。在 1000g 的离心后，上清液通过 LC-MS/MS（AB Sciex Triple Quad 5500）进行分析。

The calculation of in vitro clearance values from half-life data using hepatocytes, reflecting substrate depletion, was performed using the following equations: CL′_intrinsic [mL/(min·kg)] = (0.693/in vitro t_1/2 [min]) (liver weight [g liver/kg body mass]) (cell no. [1.1 × 10⁸]/liver weight [g])/(cell no. [1 × 10⁶]/incubation volume [mL]). The CL_blood was estimated using the nonrestricted well-stirred model: CL_blood well-stirred [L/(h·kg)] = (Q_H [L/(h·kg)]·CL′_intrinsic [L/(h·kg)])/(Q_H[L/(h·kg)] + CL′_intrinsic [L/(h·kg)]). For calculations, the following values were used: human specific liver weight of 21 g/kg body mass, hepatic blood flow of 1.32 L/(h·kg), cell number in the liver was estimated to be 1.1 × 10⁸ cells/g liver; rat specific liver weight of 32 g/kg body mass, hepatic blood flow of 4.2 L/(h·kg), cell number in the liver was estimated to be 1.1 × 10⁸ cells/g liver. (52)
使用肝细胞从半衰期数据计算体外清除值，反映底物耗竭，采用以下公式进行：CL′intrinsic [mL/(min·kg)] = (0.693/in vitro t1/2 [min]) × (肝重 [g 肝/kg 体重]) × (细胞数 [1.1 × 10^8]/肝重 [g]) / (细胞数 [1 × 10^6]/培养体积 [mL])。CLblood 使用非限制性全混模型进行估算：CLblood well-stirred [L/(h·kg)] = (QH [L/(h·kg)] × CL′intrinsic [L/(h·kg)]) / (QH[L/(h·kg)] + CL′intrinsic [L/(h·kg)])。计算中使用了以下数值：人体特异肝重为 21 g/kg 体重，肝血流量为 1.32 L/(h·kg)，肝脏中的细胞数估计为 1.1 × 10^8 个细胞/g 肝；大鼠特异肝重为 32 g/kg 体重，肝血流量为 4.2 L/(h·kg)，肝脏中的细胞数估计为 1.1 × 10^8 个细胞/g 肝。 (52)

Caco-2 Permeability Assay
Caco-2 渗透性测定

The in vitro permeation of test compounds across a Caco-2 cell monolayer, a well-established in vitro system to predict the permeability from the gastrointestinal tract, was tested according to Artursson and Karlsson. (53) Caco-2 cells (ACC 169, DSMZ, German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany) were seeded on 24-well insert plates and were allowed to grow for 14–16 d. For permeability studies, the test compounds were dissolved in DMSO and diluted to the final test concentration of 2 μM with transport buffer [Hanks’ Buffered Salt Solution, Gibco/Invitrogen, further supplemented with glucose (final concentration 19.9 mM) and HEPES (final concentration 9.8 mM)]. For determination of the apical to basolateral permeability (P_app A–B), the test compound solution was added to the apical side of the cell monolayer and transport buffer to the basolateral side of the monolayer. For determination of the basolateral to apical permeability (P_app B–A), the test compound solution was added to the basolateral side of the cell monolayer and transport buffer to the apical side of the monolayer. Samples were taken from the donor compartment at the beginning of the experiment to confirm mass balance. After an incubation of 2 h at 37 °C, samples were taken from both compartments. Samples were analyzed by LC-MS, and the apparent permeability coefficients were calculated. The efflux ratio was calculated as P_app B–A/P_app A–B. Lucifer yellow permeability was assayed for each cell monolayer to ensure cell monolayer integrity, and the permeability of atenolol (low permeability marker) and sulfasalazine (marker for active excretion) was determined for each batch as a quality control.
在体外渗透实验中，测试化合物通过 Caco-2 细胞单层（一种用于预测胃肠道渗透性的成熟体外系统）的渗透性，按照 Artursson 和 Karlsson 的方法进行测试。（53）Caco-2 细胞（ACC 169，来源于德国微生物和细胞培养物收藏中心 DSMZ，位于德国不伦瑞克）被接种在 24 孔板中，并允许其生长 14-16 天。在渗透性研究中，测试化合物溶解于 DMSO 中，并稀释至最终测试浓度 2 μM，使用转运缓冲液[汉克斯缓冲盐溶液，Gibco/Invitrogen，进一步补充葡萄糖（最终浓度 19.9 mM）和 HEPES（最终浓度 9.8 mM）]。为了确定细胞单层的顶端到基底侧渗透性（Papp A-B），将测试化合物溶液添加到细胞单层的顶端，转运缓冲液添加到基底侧。为了确定细胞单层的基底侧到顶端渗透性（Papp B-A），将测试化合物溶液添加到细胞单层的基底侧，转运缓冲液添加到顶端。实验开始时从供体室取样，以确认质量平衡。在 37°C 下孵育 2 小时后，从两个室中取样。样品通过液相色谱-质谱联用（LC-MS）进行分析，并计算了表观渗透系数。通过计算 Papp B-A/Papp A-B 得到外排比。对每个细胞单层进行了 Lucifer yellow 渗透性测定，以确保细胞单层完整性，并确定了每个批次中 atenolol（低渗透性标志物）和 sulfasalazine（活性排泄标志物）的渗透性，以进行质量控制。

Pharmacokinetic Parameters after Intravenous and Oral Application in Rats and Dogs
药代动力学参数在大鼠和狗体内静脉和口服给药后的研究

For in vivo pharmacokinetic experiments, male Wistar rats and female beagle dogs were used. Intravenous application was carried out with a species-specific plasma/DMSO formulation in rats and with a H₂O/PEG 400/EtOH formulation in dogs. Oral application in both species was by gavage with a H₂O/PEG 400/EtOH formulation. For simplification of blood drawing in rats, a silicone catheter was implanted into the right vena jugularis externa. The surgery was performed at least 1 d before substance application under isoflurane anesthesia and additional administration of an analgetic (atropine/rimadyl 3:1, 0.1 mL sc). Blood drawing (usually more than 10 time points) was done in a time window that included at least two time points after 24 h (postsubstance application). Blood was passed into heparinized tubes. Afterward, blood plasma was obtained by centrifugation at 1000g. Where necessary, the plasma was stored at −20 °C until further analysis.
在体内药代动力学实验中，使用了雄性 Wistar 大鼠和雌性比格犬。在老鼠身上，采用特定物种的血浆/DMSO 制剂进行静脉给药；在狗身上，采用 H2O/PEG 400/EtOH 制剂。两种动物均通过灌胃方式使用 H2O/PEG 400/EtOH 制剂进行口服给药。为了简化老鼠的采血，将硅胶导管植入右侧外颈静脉。手术在给药前至少 1 天进行，使用异氟醚麻醉，并额外给予镇痛剂（阿托品/瑞马唑 3:1，0.1 mL 皮下注射）。采血（通常超过 10 个时间点）在包括给药后至少 24 小时的两个时间点的时间窗口内进行。血液注入肝素化试管中。之后，通过 1000g 离心获得血浆。如有必要，将血浆储存在-20°C 直至进一步分析。

An internal standard was added to the sample, calibration, and qualifier solutions. The internal standard could also have been a compound from a different chemical class than the analyte of interest. Afterward, protein precipitation was performed by using an excess of MeCN. A buffer solution was added with a composition based on the mobile phases used in subsequent liquid chromatography. After centrifugation at 1000g, the supernatant was analyzed by LC-MS using different C18 reversed-phase columns and various mobile phase compositions. Quantification of the substance was conducted by using peak height or area calculated from extracted ion chromatograms of specific selected ion-monitoring experiments or high-resolution LC-MS experiments.
在样品、校准液和质控液中添加了内部标准。该内部标准也可以是与分析物不属于同一化学类的化合物。随后，通过使用过量的甲醇氰化物（MeCN）进行蛋白质沉淀。随后加入了一种缓冲溶液，其组成基于后续液相色谱中使用的流动相。在 1000g 的离心力下离心后，上清液通过使用不同的 C18 反相色谱柱和不同的流动相组成，由液相色谱-质谱联用（LC-MS）进行分析。通过使用从特定选择的离子监测实验或高分辨率 LC-MS 实验中提取的离子色谱图计算出的峰高或面积进行物质的定量分析。

From the plasma concentration–time course, the pharmacokinetic parameters CL (clearance), t_1/2 (terminal half-life), V_SS (volume of distribution at steady state), and F (bioavailability after oral administration) were calculated by using a validated internal pharmacokinetic calculation software.
从血浆浓度-时间曲线中，通过使用经过验证的内部药代动力学计算软件，计算了药代动力学参数 CL（清除率）、t1/2（终末半衰期）、VSS（稳态分布容积）和 F（口服后的生物利用度）。

Because substance quantification was done in plasma, the blood/plasma distribution needed to be analyzed to calculate a blood clearance value. Therefore, a defined amount of the substance was added to blood in heparinized tubes and incubated for 20 min by gently swinging. The plasma was obtained by centrifugation at 1000g. The c_blood/c_plasma value was calculated after measurement of the substance concentration in plasma and blood by using peak height or area calculated from extracted ion chromatograms of specific selected ion-monitoring experiments or high-resolution LC-MS experiments.
因为物质定量是在血浆中进行的，所以需要分析血液/血浆分布来计算血液清除率。因此，将一定量的物质添加到肝素化试管中的血液中，并轻轻摇动孵育 20 分钟。通过 1000g 的离心获得血浆。通过测量血浆和血液中物质浓度，并使用从特定选择的离子监测实验或高分辨率 LC-MS 实验中提取的离子色谱图计算出的峰高或面积来计算 cblood/cplasma 值。

Highly Purified sGC 高度纯化的 sGC

Enzyme activity was measured by the formation of [³²P]-cGMP from α-[³²P]-GTP, modified according to Hoenicka et al. (54) and Schermuly et al. (55) The modifications included using GTP, Mn²⁺/Mg²⁺, and cGMP at concentrations of 200 μM, 3 mM, and 1 mM, respectively. Enzyme concentrations were chosen carefully to achieve a substrate turnover of less than 10%, thus avoiding substrate or cofactor depletion. The characterization of the purified enzyme was performed at a protein concentration of 0.2 μg/mL. All measurements were performed in duplicate and were repeated five times. For enzyme characterization, the specific activity of sGC was expressed as x-fold stimulation vs specific basal activity. The highest DMSO concentration in the assay was 1% (v/v) and did not elicit any effect per se on cGMP production.
酶活性通过α-[32P]-GTP 形成[32P]-cGMP 来测定，方法参照 Hoenicka 等（54）和 Schermuly 等（55）的研究，并进行了修改。修改内容包括使用 GTP、Mn2+/Mg2+和 cGMP，其浓度分别为 200 μM、3 mM 和 1 mM。酶浓度选择得非常谨慎，以实现底物转换率低于 10%，从而避免底物或辅因子耗竭。纯化酶的特性分析在蛋白质浓度为 0.2 μg/mL 时进行。所有测量均重复两次，并重复五次。对于酶的特性分析，sGC 的比活性以倍数表示，与特定基础活性相比。实验中 DMSO 的最高浓度为 1%（体积比），本身对 cGMP 的产生没有影响。

Recombinant sGC-Overexpressing Cell Line
重组 sGC 高表达细胞系

The cellular activity of the test compounds was determined using a recombinant sGC-overexpressing cell line, as previously described. (34) Briefly, cells were plated in a volume of 25 μL on white 384-well Greiner Bio-One microplates and were cultured for 1 or 2 d in medium. Medium was removed, and cells were loaded for 3 h with calcium-free Tyrode-containing coelenterazine. Serial dilutions of the test compounds in a volume of 10 μL in calcium-free Tyrode were applied to the cells for 6 min. Thereafter, 35 μL of Tyrode-containing calcium (final concentration: 3 mM) was added to the cells and the emitted light was measured for 40 s using a CCD camera in a light-tight box. The minimal effective concentration (MEC) was determined as the concentration where a ≥3-fold increase in the basal luminescence value was observed.
细胞活性测试化合物的活性通过重组 sGC 过表达细胞系进行测定，方法如前所述。（34）简要来说，细胞以 25 μL 的体积接种于白色 384 孔 Greiner Bio-One 微孔板中，并在培养基中培养 1 或 2 天。移除培养基后，细胞用无钙 Tyrode 培养基和海葵素共培养 3 小时。随后，将测试化合物在 10 μL 无钙 Tyrode 培养基中进行系列稀释，并作用于细胞 6 分钟。之后，向细胞中加入含钙 Tyrode 培养基（终浓度：3 mM）35 μL，并使用密封箱中的 CCD 相机测量发射的光 40 秒。最小有效浓度（MEC）被定义为观察到基线荧光值增加≥3 倍时的浓度。

Isolated Vessels and Tolerance
孤立船舶与容忍

The relaxing effects of 24 on aortas, saphenous arteries, coronary arteries, and veins, as well as the investigation on aortic rings taken from either normal or nitrate-tolerant rabbits, were performed as previously described. (39)
24 对主动脉、隐静脉、冠状动脉和静脉的放松作用，以及从正常或耐受硝酸盐的兔子身上取出的主动脉环的研究，均按先前所述进行。（39）

Rat Heart Langendorff Preparation
大鼠心脏 Langendorff 制备

Male Wistar rats (200–250 g) were anesthetized using Narcoren (100 mg/kg ip). The heart was rapidly excised and connected to a Langendorff perfusion system (FMI GmbH, Seeheim-Ober Beerbach, Germany). The heart was perfused at a constant flow rate of 10 mL/min with Krebs–Henseleit buffer solution equilibrated with 95% O₂ and 5% CO₂. The perfusion solution contained (in mmol/L): NaCl 118, KCl 3, NaHCO₃ 22, KH₂PO₄ 1.2, MgSO₄ 1.2, CaCl₂ 1.8, glucose 10, and sodium pyruvate 2. A pressure transducer registered the perfusion pressure in the system. The left ventricular pressure was measured using a second pressure transducer connected to a water-filled balloon which was inserted into the left ventricle via the left atrium. The end diastolic pressure was initially set to 8–10 mmHg by adjusting the volume of the balloon. The hearts were spontaneously beating. The signals from the pressure transducer were amplified, registered, and used for the calculation of the heart frequency and + dP/dt_max by a personal computer. 24 was dissolved in a mixture of 10% DMSO and 90% saline and infused for 20 min with increasing concentration steps into the aortic cannula at a rate of 1% of the total flow rate. All values are presented as relative changes of baseline values before compound application.
雄性 Wistar 大鼠（体重 200-250 克）使用纳可伦（100 毫克/千克，ip）进行麻醉。迅速取出心脏并将其连接到 Langendorff 灌流系统（德国 FMI GmbH，Seeheim-Ober Beerbach）。心脏以每分钟 10 毫升的恒定流速用 Krebs-Henseleit 缓冲液灌流，该缓冲液与 95%氧气和 5%二氧化碳平衡。灌流液成分（以毫摩尔/升计）：NaCl 118，KCl 3，NaHCO3 22，KH2PO4 1.2，MgSO4 1.2，CaCl2 1.8，葡萄糖 10，和钠丙酮酸 2。一个压力传感器记录系统中的灌流压力。使用连接到水充气囊的第二压力传感器测量左心室压力，该气囊通过左心房插入左心室。初始将舒张末期压力设置为 8-10 毫米汞柱，通过调整气囊的体积来实现。心脏自发跳动。压力传感器的信号经过放大、记录，并由个人计算机用于计算心率及+ dP/dtmax。 24%的溶液溶解在 10%的二甲基亚砜和 90%的盐水中，以总流量的 1%的速率，通过增加浓度梯度，在主动脉导管中持续输注 20 分钟。所有数值均以化合物应用前的基础值相对变化表示。

Chronic Treatment Study with L-NAME-Treated Renin Transgenic Rats
慢性 L-NAME 处理肾素转基因大鼠治疗研究

Fifty male renin transgenic rats carrying an additional mouse renin gene [RenTG(mRRen2)27] at the age of 8 weeks were used. L-NAME was chronically administered via the drinking water (50 mg/L) in all study groups. Animals were randomly allocated to three study groups: placebo (control) (n = 20), 24 low dose, and 24 high dose (3 and 10 mg/kg per day, respectively, administered po by gavage qd, n = 15 per group). Blood pressure was measured via the tail-cuff method once before the start of the study (day 0) to exclude preexisting differences between the groups and on day 7, 14, and 21. Body weight and survival were assessed on day 1, 8, and 15 and at the study end. At the end of the study (day 22), all animals were anesthetized, blood was collected, and animals were sacrificed; blood was taken in order to assess plasma parameters, and the heart was dissected into the left and right ventricles and was weighed to assess potential heart hypertrophy. Creatinine, urea, and renin activity in plasma were determined after extraction, as previously described. (39, 56)
在 8 周龄时，使用了 50 只携带额外小鼠肾素基因[RenTG(mRRen2)27]的雄性肾素转基因大鼠。所有研究组均通过饮用水（50 mg/L）慢性给予 L-NAME。动物被随机分配到三个研究组：安慰剂（对照组）（n = 20）、24 只低剂量组和 24 只高剂量组（分别每天口服灌胃 3 mg/kg 和 10 mg/kg，每组 n = 15）。血压在研究开始前（第 0 天）和第 7 天、14 天和 21 天通过尾动脉血压计法测量，以排除各组之间存在的先存差异。体重和存活情况在第 1 天、第 8 天、第 15 天和研究结束时进行评估。在研究结束时（第 22 天），所有动物均被麻醉，采集血液，并进行安乐死；血液采集用于评估血浆参数，心脏被解剖成左心室和右心室，并称重以评估潜在的心脏肥大。血浆中的肌酐、尿素和肾素活性在提取后按先前描述的方法测定。（39，56）

Statistics 统计

The unpaired t test was used to detect significant differences between the groups of interest. Results (mean ± SEM) were considered significant when the probability error (P) was less than 0.05, 0.01,and 0.001 for *, **, and ***, respectively.
配对样本 t 检验用于检测感兴趣组之间的显著差异。结果（均值±标准误）认为当概率误差（P）小于 0.05、0.01 和 0.001 时，分别表示为*、**和***，认为结果具有统计学意义。

Supporting Information 支持信息

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The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jmedchem.7b00449.
支持信息可在 ACS 出版物网站上免费获取，网址为 DOI：10.1021/acs.jmedchem.7b00449。

¹H NMR spectra of selected compounds 3, 21, 22, and 24; CYP-inhibition results of 24 (PDF)
1H NMR 光谱图选自化合物 3、21、22 和 24；化合物 24 的 CYP 抑制结果（PDF）
Molecular formula strings (CSV)
分子式字符串（CSV）

Discovery of the Soluble Guanylate Cyclase Stimulator Vericiguat (BAY 1021189) for the Treatment of Chronic Heart Failure

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Discovery of the Soluble Guanylate Cyclase Stimulat
发现可溶性鸟苷酸环化酶激动剂

or Vericiguat (BAY 1021189) for the
或维利吉单抗（BAY 1021189）用于

Treatment of Chronic Heart Failure
慢性心力衰竭的治疗

Markus Follmann,* Jens Ackerstaff, Gorden Redlich,
马库斯·福尔曼、* 詹斯·阿克斯塔夫、戈登·雷德里希，

Frank Wunder, Dieter Lang, Armin
弗兰克·温德，迪特·朗，阿明

Kern, Peter Fey, Nils Griebenow, Walter Kroh, Eva-M
凯恩，彼得·费伊，尼尔·格里本诺，瓦尔特·克罗，伊娃-玛

aria Becker-Pelster, Axel Kretschmer,
贝克尔-佩尔斯特，阿克斯·克雷施默

Volker Geiss, Volkhart Min-Jian Li, Alexander Strau
Volker Geiss，Volkhart Min-Jian Li，Alexander Strau

b, Joachim Mittendorf, Rolf Jautelat,
b, 耶奥希姆·米特多夫，罗尔夫·贾特拉特，

Hartmut Schirok, Karl-Heinz Schlemmer, Klemens Lust
哈特穆特·施里罗克，卡尔-海因茨·施莱默，克莱门斯·卢斯特

ig, Michael Gerisch, Andreas Knorr,
""" ig，迈克尔·格里斯奇，安德烈亚斯·诺尔， """

Hanna Tinel, Thomas Mondritzki, Hubert Trübel, Pete
汉娜·蒂内尔，托马斯·蒙德里茨基，胡伯特·特鲁贝尔，皮特

r Sandner, and Johannes-Peter Stasch
桑德纳，约翰内斯-彼得·斯塔什

Bayer AG, Drug Discovery, Aprather Weg 18a, 42113 W
拜耳股份公司，药物发现，阿帕瑟尔韦格 18a，42113 W 区

uppertal, Germany 乌普塔尔，德国

*Corresponding author: 对应作者：

Dr. Markus Follmann
马库斯·福尔曼博士

E-mail: 电子邮件：

markus.follmann@bayer.com

Supporting information 支持信息

NMR spectra of selected compounds
核磁共振谱图选定的化合物

Table S6 表 S6

: CYP-inhibition results of vericiguat
维立西格他尔的 CYP 抑制结果

H NMR of compound
化合物核磁共振氢谱

H NMR of compound

Terms & Conditions 《服务条款》

Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html.
大多数电子辅助信息文件无需订阅 ACS Web Editions 即可获取。这些文件可以按文章下载用于研究（如果相关文章链接了公共使用许可，该许可可能允许其他用途）。如需其他用途，可通过 RightsLink 许可系统向 ACS 申请许可：http://pubs.acs.org/page/copyright/permissions.html。

Author Information 作者信息

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Corresponding Author 对应作者
- Markus Follmann - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany; http://orcid.org/0000-0003-1246-3603; Email: markus.follmann@bayer.com
  马库斯·福尔曼 - 药物发现，拜耳公司，德国乌珀塔尔市阿帕瑟尔韦格 18a 号，42113； http://orcid.org/0000-0003-1246-3603；邮箱：markus.follmann@bayer.com
Authors 作者
- Jens Ackerstaff - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Gorden Redlich - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Frank Wunder - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Dieter Lang - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Armin Kern - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Peter Fey - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Nils Griebenow - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Walter Kroh - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Eva-Maria Becker-Pelster - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Axel Kretschmer - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Volker Geiss - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Volkhart Li - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Alexander Straub - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Joachim Mittendorf - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Rolf Jautelat - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Hartmut Schirok - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Karl-Heinz Schlemmer - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Klemens Lustig - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Michael Gerisch - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Andreas Knorr - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Hanna Tinel - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Thomas Mondritzki - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Hubert Trübel - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Peter Sandner - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
- Johannes-Peter Stasch - Drug Discovery, Bayer AG, Aprather Weg 18a, 42113 Wuppertal, Germany
Author Contributions
The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.
Notes
The authors declare the following competing financial interest(s): All authors are or have been employees of Bayer AG.

Acknowledgment

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Picture used for abstract/Table of Contents graphic taken from “Unseen Extremes: Mapping the World’s Greatest Mountains” by courtesy of Deutsches Zentrum für Luft- und Raumfahrt eV.

Abbreviations Used

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sGC	soluble guanylate cyclase
cGMP	cyclic guanosine 5′-monophosphate
HMDS	hexamethyldisilazane
MEC	minimum effective concentration
ClogD	calculated logarithm of distribution coefficient
Cl_b	blood clearance

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This article references 56 other publications.

1
Evgenov, O. V.; Pacher, P.; Schmidt, P. M.; Hasko, G.; Schmidt, H. H. H. W.; Stasch, J.-P. NO-independent stimulators and activators of soluble guanylate cyclase: discovery and therapeutic potential Nat. Rev. Drug Discovery 2006, 5, 755– 768 DOI: 10.1038/nrd2038
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NO-independent stimulators and activators of soluble guanylate cyclase: discovery and therapeutic potential
Evgenov, Oleg V.; Pacher, Pal; Schmidt, Peter M.; Hasko, Gyoergy; Schmidt, Harald H. H. W.; Stasch, Johannes-Peter
Nature Reviews Drug Discovery (2006), 5 (9), 755-768CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)
A review. Sol. guanylate cyclase (sGC) is a key signal-transduction enzyme activated by nitric oxide (NO). Impaired bioavailability and/or responsiveness to endogenous NO has been implicated in the pathogenesis of cardiovascular and other diseases. Current therapies that involve the use of org. nitrates and other NO donors have limitations, including non-specific interactions of NO with various biomols., lack of response and the development of tolerance following prolonged administration. Compds. that activate sGC in an NO-independent manner might therefore provide considerable therapeutic advantages. Here we review the discovery, biochem., pharmacol. and clin. potential of heme-dependent sGC stimulators (including YC-1, BAY 41-2272, BAY 41-8543, CFM-1571 and A-350619) and heme-independent sGC activators (including BAY 58-2667 and HMR-1766).
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Follmann, M.; Griebenow, N.; Hahn, M. G.; Hartung, I.; Mais, F.-J.; Mittendorf, J.; Schaefer, M.; Schirok, H.; Stasch, J.-P.; Stoll, F.; Straub, A. The chemistry and biology of soluble guanylate cyclase stimulators and activators Angew. Chem., Int. Ed. 2013, 52, 9442– 9462 DOI: 10.1002/anie.201302588
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2
The Chemistry and Biology of Soluble Guanylate Cyclase Stimulators and Activators
Follmann, Markus; Griebenow, Nils; Hahn, Michael G.; Hartung, Ingo; Mais, Franz-Josef; Mittendorf, Joachim; Schaefer, Martina; Schirok, Hartmut; Stasch, Johannes-Peter; Stoll, Friederike; Straub, Alexander
Angewandte Chemie, International Edition (2013), 52 (36), 9442-9462CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
A review. The vasodilatory properties of nitric oxide (NO) have been utilized in pharmacotherapy for more than 130 years. Still today, NO-donor drugs are important in the management of cardiovascular diseases. However, inhaled NO or drugs releasing NO and org. nitrates are assocd. with noteworthy therapeutic shortcomings, including resistance to NO in some disease states, the development of tolerance during long-term treatment, and nonspecific effects, such as post-translational modification of proteins. The beneficial actions of NO are mediated by stimulation of sol. guanylate cyclase (sGC), a heme-contg. enzyme which produces the intracellular signaling mol. cGMP. Recently, two classes of compds. have been discovered that amplify the function of sGC in a NO-independent manner, the so-called sGC stimulators and sGC activators. The most advanced drug, the sGC stimulator riociguat, has successfully undergone Phase III clin. trials for different forms of pulmonary hypertension.
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Cerra, M. C.; Pellegrino, D. Cardiovascular cGMP-generating systems in physiological and pathological conditions Curr. Med. Chem. 2007, 14, 585– 599 DOI: 10.2174/092986707780059715
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3
Cardiovascular cGMP-generating systems in physiological and pathological conditions
Cerra, M. C.; Pellegrino, D.
Current Medicinal Chemistry (2007), 14 (5), 585-599CODEN: CMCHE7; ISSN:0929-8673. (Bentham Science Publishers Ltd.)
A review. The intracellular messenger cyclic GMP (cGMP) represents the key signal in several transduction pathways throughout the animal world. In the heart cGMP signaling contributes to functional interaction of different cell types. Nitric oxide (NO) and natriuretic peptides (NPs), major autocrine-paracrine cardiovascular regulators, increment intracellular cGMP through guanylate cyclases (GCs). NO and NPs interact with two GC types: cytosolic (sol.: sGC) and membrane bound [particulate: pGC (NP receptor types A and B)], resp. Depending on sub-cellular localization and regulation of the enzymes, cGMP produced by either pGC or sGC exerts different complementary effects. The two pathways are reciprocally regulated. NPs-depending pGC is modulated by NO-cGMP signaling, and the activity of NO is influenced by cellular concns. of both NO itself and NPs. This heterologous feedback regulates GCs, linking cardiovascular autocrine-paracrine activities of NPs and NO. Importance of these cGMP converging routes goes far beyond their role under normal conditions. They are of relevance esp. in disease states when tissue and circulating levels of NPs, and local NO prodn. are altered. An example is the endothelial dysfunction assocd. with deficient NO prodn. and uncoupled endothelium-myocardium communications. In this case, NPs-pGC-cGMP could supplement the reduced activity of NO-scGC-cGMP pathway. In addn., these systems regulate cell growth and apoptosis, playing a role in myocardial pathol. morpho-functional remodeling. Here we will review recent concepts on NO/NPs dependent control of heart function in vertebrates, also focusing on cGMP-activated downstream signaling and its role in health and disease conditions.
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Gladwin, M. T. Deconstructing endothelial dysfunction: soluble guanylyl cyclase oxidation and the NO resistance syndrome J. Clin. Invest. 2006, 116, 2330– 2332 DOI: 10.1172/JCI29807
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4
Deconstructing endothelial dysfunction: soluble guanylyl cyclase oxidation and the NO resistance syndrome
Gladwin, Mark T.
Journal of Clinical Investigation (2006), 116 (9), 2330-2332CODEN: JCINAO; ISSN:0021-9738. (American Society for Clinical Investigation)
A review. In this issue of the JCI, Stasch and colleagues suggest that a novel drug, BAY 58-2667, potently activates a pool of oxidized and heme-free sol. guanylyl cyclase (sGC; see the related article beginning on page 2552). The increased vasodilatory potency of BAY 58-2667 the authors found in a no. of animal models of endothelial dysfunction and in human blood vessels from patients with diabetes suggests that there exists a subphenotype of endothelial dysfunction characterized by receptor-level NO resistance. Diseases assocd. with NO resistance would appear to be ideally suited for therapies directed at restoring redox homeostasis, sGC activity, and NO sensitivity.
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Hoenicka, M.; Schmid, C. Cardiovascular effects of modulators of soluble guanylyl cyclase activity Cardiovasc. Hematol. Agents Med. Chem. 2008, 6, 287– 301 DOI: 10.2174/187152508785909555
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5
Cardiovascular effects of modulators of soluble guanylyl cyclase activity
Hoenicka, Markus; Schmid, Christof
Cardiovascular & Hematological Agents in Medicinal Chemistry (2008), 6 (4), 287-301CODEN: CHAAA5; ISSN:1871-5257. (Bentham Science Publishers Ltd.)
A review. Sol. guanylyl cyclase (sGC) is one of the key enzymes of the nitric-oxide (NO)/cyclic 3',5'-guanosine monophosphate (cGMP) pathway. Located in virtually all mammalian cells, it controls the vessel tone, smooth muscle cell growth, platelet aggregation, and leukocyte adhesion. In vivo sGC activity is mainly regulated by NO which in turn is released from L-arginine by nitric oxide synthases. One of the main diseases of the cardiovascular system, endothelial dysfunction, leads to a diminished NO synthesis and thus increases vessel tone as well as the risk of thrombosis. The predominant therapeutic approach to this condition is a NO replacement therapy, as exemplified by org. nitrates, molsidomin, and other NO releasing substances. Recent advances in drug discovery provided a variety of other approaches to activate sGC, which may help to circumvent both the tolerance problem and some non-specific actions assocd. with NO donor drugs. Substances like BAY 41-2272 stimulate sGC in a heme-dependent fashion and synergize with NO, allowing to enhance the effects both of endogenous NO and of exogenous NO donors. On the other hand, heme-independent activators like BAY 58-2667 allow to activate sGC even if it is rendered unresponsive to NO due to oxidative stress or heme loss. Furthermore, a few substances have been described as specific inhibitors of sGC that allow to alleviate the effects of excess NO prodn. as seen in shock. This review discusses the cardiovascular effects of heme-dependent and heme-independent activators as well as of inhibitors of sGC.
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Priviero, F. B.; Webb, R. C. Heme-dependent and independent soluble guanylate cyclase activators and vasodilation J. Cardiovasc. Pharmacol. 2010, 56, 229– 233 DOI: 10.1097/FJC.0b013e3181eb4e75
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Heme-Dependent and Independent Soluble Guanylate Cyclase Activators and Vasodilation
Priviero, Fernanda B. M.; Webb, R. Clinton
Journal of Cardiovascular Pharmacology (2010), 56 (3), 229-233CODEN: JCPCDT; ISSN:0160-2446. (Lippincott Williams & Wilkins)
A review. Since the discovery of nitric oxide (NO), which is released from endothelial cells as the main mediator of vasodilation, its target, the sol. guanylyl cyclase (sGC), has become a focus of interest for the treatment of diseases assocd. with endothelial dysfunction. NO donors were developed to suppress NO deficiency; however, tolerance to org. nitrates was reported. Non-NO-based drugs targeting sGC were developed to overcome the problem of tolerance. In this review, we briefly describe the process of sGC activation by its main physiol. activator NO and the advances in the development of drugs capable of activating sGC in a NO-independent manner. sGC stimulators, as some of these drugs are called, require the integrity of the reduced heme moiety of the prosthetic group within the sGC and therefore are called heme-dependent stimulators. Other drugs are able to activate sGC independent of heme moiety and are hence called heme-independent activators. Because pathol. conditions modulate sGC and oxidize the heme moiety, the heme-independent sGC activators could potentially become drugs of choice because of their higher affinity to the oxidized enzyme. However, these drugs are still undergoing clin. trials and are not available for clin. use.
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Packer, C. S. Soluble guanylate cyclase (sGC) down-regulation by abnormal extracellular matrix proteins as a novel mechanism in vascular dysfunction: implications in metabolic syndrome Cardiovasc. Res. 2006, 69, 302– 303 DOI: 10.1016/j.cardiores.2005.12.006
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Soluble guanylate cyclase (sGC) down-regulation by abnormal extracellular matrix proteins as a novel mechanism in vascular dysfunction: Implications in metabolic syndrome
Packer, C. Subah
Cardiovascular Research (2006), 69 (2), 302-303CODEN: CVREAU; ISSN:0008-6363. (Elsevier B.V.)
A review. The role of endothelial NO prodn., accumulation of extracellular matrix proteins and altered activity of sol. guanylate cyclase in endothelial dysfunction and metabolic syndrome is discussed.
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Mayer, B.; Koesling, D. cGMP signalling beyond nitric oxide Trends Pharmacol. Sci. 2001, 22, 546– 548 DOI: 10.1016/S0165-6147(00)01889-7
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8
cGMP signalling beyond nitric oxide
Mayer, Bernd; Koesling, Doris
Trends in Pharmacological Sciences (2001), 22 (11), 546-548CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Science Ltd.)
A review. Many of the physiol. effects of nitric oxide are mediated by activation of sol. guanylyl cyclase, resulting in cellular cGMP accumulation. In the 1990s, the benzylindazole deriv. YC-1 was identified as a novel modulator of cGMP signaling that exerted complex actions in a NO-independent manner. A recent study describes a high-affinity YC-1 analog that decreases blood pressure in hypertensive rats and increases bleeding time, which suggests that this drug might have therapeutic potential as a vasodilator with antiplatelet activity.
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Nioche, P.; Berka, V.; Vipond, J.; Minton, N.; Tsai, A. L.; Raman, C. S. Femtomolar sensitivity of a NO sensor from Clostridium botulinum Science 2004, 306, 1550– 1553 DOI: 10.1126/science.1103596
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9
Femtomolar sensitivity of a NO sensor from Clostridium botulinum
Nioche, Pierre; Berka, Vladimir; Vipond, Julia; Minton, Nigel; Tsai, Ah-Lim; Raman, C. S.
Science (Washington, DC, United States) (2004), 306 (5701), 1550-1553CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
Nitric oxide (NO) is extremely toxic to Clostridium botulinum, but its mol. targets are unknown. Here, the authors identify a heme protein sensor (SONO) that displays femtomolar affinity for NO. The crystal structure of the SONO heme domain reveals a previously undescribed fold and a strategically placed tyrosine residue that modulates heme-nitrosyl coordination. Furthermore, the domain architecture of a SONO ortholog cloned from Chlamydomonas reinhardtii indicates that NO signaling through cGMP arose before the origin of multicellular eukaryotes. The authors' findings have broad implications for understanding bacterial responses to NO, as well as for the activation of mammalian NO-sensitive guanylyl cyclase.
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Pellicena, P.; Karow, D. S.; Boon, E. M.; Marletta, M. A.; Kuriyan, J. Crystal structure of an oxygen-binding heme domain related to soluble guanylate cyclases Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 12854– 12859 DOI: 10.1073/pnas.0405188101
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Crystal structure of an oxygen-binding heme domain related to soluble guanylate cyclases
Pellicena, Patricia; Karow, David S.; Boon, Elizabeth M.; Marletta, Michael A.; Kuriyan, John
Proceedings of the National Academy of Sciences of the United States of America (2004), 101 (35), 12854-12859CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
Sol. guanylate cyclases are nitric oxide-responsive signaling proteins in which the nitric oxide sensor is a heme-binding domain of unknown structure that we have termed the heme-NO and oxygen binding (H-NOX) domain. H-NOX domains are also found in bacteria, either as isolated domains, or are fused through a membrane-spanning region to methyl-accepting chemotaxis proteins. We have detd. the crystal structure of an oxygen-binding H-NOX domain of one such signaling protein from the obligate anaerobe Thermoanaerobacter tengcongensis at 1.77-Å resoln., revealing a protein fold unrelated to known structures. Particularly striking is the structure of the protoporphyrin IX group, which is distorted from planarity to an extent not seen before in protein-bound heme groups. Comparison of the structure of the H-NOX domain in two different crystal forms suggests a mechanism whereby alteration in the degree of distortion of the heme group is coupled to changes on the mol. surface of the H-NOX domain and potentially to changes in intermol. interactions.
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Wedel, B.; Humbert, P.; Harteneck, C.; Foerster, J.; Malkewitz, J.; Bohme, E.; Schultz, G.; Koesling, D. Mutation of His-105 in the β₁ subunit yields a nitric oxide-insensitive form of soluble guanylyl cyclase Proc. Natl. Acad. Sci. U. S. A. 1994, 91, 2592– 2596 DOI: 10.1073/pnas.91.7.2592
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Mutation of His-105 in the β1 subunit yields a nitric oxide-insensitive form of soluble guanylyl cyclase
Wedel, Barbara; Humbert, Peter; Harteneck, Christian; Foerster, John; Malkewitz, Jurgen; Boehme, Eycke; Schultz, Gunter; Koesling, Doris
Proceedings of the National Academy of Sciences of the United States of America (1994), 91 (7), 2592-6CODEN: PNASA6; ISSN:0027-8424.
Sol. guanylyl cyclase [GTP pyrophosphate-lyase (cyclizing); EC 4.6.1.2] is a hemoprotein that exists as a heterodimer; the heme moiety has been proposed to bind nitric oxide, resulting in a dramatic activation of the enzyme. Mutation of six conserved His residues reduced but did not abolish nitric oxide stimulation whereas a change of His-105 to Phe in the β1 subunit yielded a heterodimer that retained basal cyclase activity but failed to respond to nitric oxide. Heme was not detected as a component of the mutant heterodimer and protophorphyrin IX failed to stimulate enzyme activity. The activity of the His mutant was almost identical to that of the wild-type enzyme in the presence of KCN, suggesting that disruption of heme binding is the principal effect of the mutation. Thus, the mutation provides a means to inhibit the nitric oxide-sensitive guanylyl cyclase signalling pathway.
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Zabel, U.; Hausler, C.; Weeger, M.; Schmidt, H. H. Homodimerization of soluble guanylyl cyclase subunits. Dimerization analysis using a glutathione S-transferase affinity tag J. Biol. Chem. 1999, 274, 18149– 18152 DOI: 10.1074/jbc.274.26.18149
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Homodimerization of soluble guanylyl cyclase subunits. Dimerization analysis using a glutathione S-transferase affinity tag
Zabel, Ulrike; Hausler, Christoph; Weeger, Monika; Schmidt, Harald H. H. W.
Journal of Biological Chemistry (1999), 274 (26), 18149-18152CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
Sol. guanylyl cyclase (sGC) is an α/β-heterodimeric hemoprotein that, upon interaction with the intercellular messenger mol. NO, generates cGMP. Although the related family of particulate guanylyl cyclases (pGCs) forms active homodimeric complexes, it is not known whether homodimerization of sGC subunits occurs. We report here the expression in Sf9 cells of glutathione S-transferase-tagged recombinant human sGCα1 and β1 subunits, applying a novel and rapid purifn. method based on GSH-Sepharose affinity chromatog. Surprisingly, in intact Sf9 cells, both homodimeric GSTα/α and GSTβ/β complexes were formed that were catalytically inactive. Upon coexpression of the resp. complementary subunits, GSTα/β or GSTβ/α heterodimers were preferentially formed, whereas homodimers were still detectable. When subunits were mixed after expression, e.g. GSTβ and β or GSTα and β, no dimerization was obsd. In conclusion, our data suggest the previously unrecognized possibility of a physiol. equil. between homo- and heterodimeric sGC complexes.
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Zabel, U.; Weeger, M.; La, M.; Schmidt, H. H. W. Human soluble guanylate cyclase: functional expression and revised isoenzyme family Biochem. J. 1998, 335, 51– 57 DOI: 10.1042/bj3350051
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Human soluble guanylate cyclase: functional expression and revised isoenzyme family
Zabel, Ulrike; Weeger, Monika; La, Mylinh; Schmidt, Harald H. H. W.
Biochemical Journal (1998), 335 (1), 51-57CODEN: BIJOAK; ISSN:0264-6021. (Portland Press Ltd.)
Sol. guanylate cyclase (sGC), a heterodimeric (α/β) heme protein that converts GTP to the second messenger cGMP, functions as the receptor for nitric oxide (NO) and nitrovasodilator drugs. Three distinct cDNA species of each subunit (α1-α3, β1-β3) have been reported from various species. From human sources, none of these have been expressed as functionally active enzyme. Here the authors described the expression of human α/β heterodimeric sGC in Sf9 cells yielding active recombinant enzyme that was stimulated by the nitrovasodilator sodium nitroprusside or the NO-independent activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1). At the protein level, both α and β subunits were detected in human tissues, suggesting co-expression also in vivo. Moreover, resequencing of the human cDNA clones [originally termed α3 and β3] revealed several sequencing errors in human α3; correction of these eliminated major regions of divergence from rat and bovine α1. As human β3 also displays more than 98% similarity to rat and bovine β1 at the amino acid level, α3 and β3 represent the human homologs of rat and bovine α1 and β1, and the isoenzyme family is decreased to two isoforms for each subunit (α1, α2; β1, β2). Having access to the human key enzyme of NO signaling will now permit the study of novel sGC-modulating compds. with therapeutic potential.
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Murad, F. Shattuck Lecture. Nitric oxide and cyclic GMP in cell signaling and drug development N. Engl. J. Med. 2006, 355, 2003– 2011 DOI: 10.1056/NEJMsa063904
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Nitric oxide and cyclic GMP in cell signaling and drug development
Murad, Ferid
New England Journal of Medicine (2006), 355 (19), 2003-2011CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
A review. The topics discussed include: guanylyl cyclase activity; endothelial activity; nitric oxide synthase; activation of sol. guanylyl cyclase; steady-state levels of cGMP; and cGMP signaling in drug development.
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Stasch, J. P.; Hobbs, A. J. NO-independent, haem-dependent soluble guanylate cyclase stimulators Handb. Exp. Pharmacol. 2009, 191, 277– 308 DOI: 10.1007/978-3-540-68964-5_13
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NO-independent, haem-dependent soluble guanylate cyclase stimulators
Stasch, Johannes-Peter; Hobbs, Adrian J.
Handbook of Experimental Pharmacology (2009), 191 (cGMP: Generators, Effectors and Therapeutic Implications), 277-308CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
A review. The nitric oxide (NO) signalling pathway is altered in cardiovascular diseases, including systemic and pulmonary hypertension, stroke, and atherosclerosis. The vasodilatory properties of NO have been exploited for over a century in cardiovascular disease, but NO donor drugs and inhaled NO are assocd. with significant shortcomings, including resistance to NO in some disease states, the development of tolerance during long-term treatment, and non-specific effects such as post-translational modification of proteins. The development of pharmacol. agents capable of directly stimulating the NO receptor, sol. guanylate cyclase (sGC), is therefore highly desirable. The benzylindazole compd. YC-1 was the first sGC stimulator to be identified; this compd. formed a lead structure for the development of optimized sGC stimulators with improved potency and specificity for sGC, including CFM-1571, BAY 41-2272, BAY 41-8543, and BAY 63-2521. In contrast to the NO- and haem-independent sGC activators such as BAY 58-2667, these compds. stimulate sGC activity independent of NO and also act in synergy with NO to produce anti-aggregatory, anti-proliferative, and vasodilatory effects. Recently, aryl-acrylamide compds. were identified independent of YC-1 as sGC stimulators; although structurally dissimilar to YC-1, they have a similar mode of action and promote smooth muscle relaxation. Pharmacol. stimulators of sGC may be beneficial in the treatment of a range of diseases, including systemic and pulmonary hypertension, heart failure, atherosclerosis, erectile dysfunction, and renal fibrosis. An sGC stimulator, BAY 63-2521, is currently in clin. development as an oral therapy for patients with pulmonary hypertension. It has demonstrated efficacy in a proof-of-concept study, reducing pulmonary vascular resistance and increasing cardiac output from baseline. A full, phase 2 trial of BAY 63-2521 in pulmonary hypertension is underway.
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Schmidt, H. H.; Schmidt, P. M.; Stasch, J. P. NO- and haem-independent soluble guanylate cyclase activators Handb. Exp. Pharmacol. 2009, 191, 309– 339 DOI: 10.1007/978-3-540-68964-5_14
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NO- and haem-independent soluble guanylate cyclase activators
Schmidt, Harald H. H. W.; Schmidt, Peter M.; Stasch, Johannes-Peter
Handbook of Experimental Pharmacology (2009), 191 (cGMP: Generators, Effectors and Therapeutic Implications), 309-339CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
A review. Oxidative stress, a risk factor for several cardiovascular disorders, interferes with the NO/sGC/cGMP signalling pathway through scavenging of NO and formation of the strong intermediate oxidant, peroxynitrite. Under these conditions, endothelial and vascular dysfunction develops, culminating in different cardio-renal and pulmonary-vascular diseases. Substituting NO with org. nitrates that release NO (NO donors) has been an important principle in cardiovascular therapy for more than a century. However, the development of nitrate tolerance limits their continuous clin. application and, under oxidative stress and increased formation of peroxynitrite foils the desired therapeutic effect. To overcome these obstacles of nitrate therapy, direct NO- and haem-independent sGC activators have been developed, such as BAY 58-2667 (cinaciguat) and HMR1766 (ataciguat), showing unique biochem. and pharmacol. properties. Both compds. are capable of selectively activating the oxidized/haem-free enzyme via binding to the enzyme's haem pocket, causing pronounced vasodilatation. The potential importance of these new drugs resides in the fact that they selectively target a modified state of sGC that is prevalent under disease conditions as shown in several animal models and human disease. Activators of sGC may be beneficial in the treatment of a range of diseases including systemic and pulmonary hypertension (PH), heart failure, atherosclerosis, peripheral arterial occlusive disease (PAOD), thrombosis and renal fibrosis. The sGC activator HMR1766 is currently in clin. development as an oral therapy for patients with PAOD. The sGC activator BAY 58-2667 has demonstrated efficacy in a proof-of-concept study in patients with acute decompensated heart failure (ADHF), reducing pre- and afterload and increasing cardiac output from baseline. A phase IIb clin. study for the indication of ADHF is currently underway.
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Stasch, J.-P.; Evgenov, O. V. Soluble guanylate cyclase stimulators in pulmonary hypertension Handb. Exp. Pharmacol. 2013, 218, 279– 313 DOI: 10.1007/978-3-662-45805-1_12
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Soluble Guanylate Cyclase Stimulators in Pulmonary Hypertension
Stasch, Johannes-Peter; Evgenov, Oleg V.
Handbook of Experimental Pharmacology (2013), 218 (Pharmacotherapy of Pulmonary Hypertension), 279-313CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
A review. Sol. guanylate cyclase (sGC) is a key enzyme in the nitric oxide (NO) signalling pathway. On binding of NO to its prosthetic heme group, sGC catalyzes the synthesis of the second messenger cyclic guanosine monophosphate (cGMP), which promotes vasodilation and inhibits smooth muscle proliferation, leukocyte recruitment, platelet aggregation and vascular remodelling through a no. of downstream mechanisms. The central role of the NO-sGC-cGMP pathway in regulating pulmonary vascular tone is demonstrated by the dysregulation of NO prodn., sGC activity and cGMP degrdn. in pulmonary hypertension (PH). The sGC stimulators are novel pharmacol. agents that directly stimulate sGC, both independently of NO and in synergy with NO. Optimization of the first sGC stimulator, YC-1, led to the development of the more potent and more specific sGC stimulators, BAY 41-2272, BAY 41-8543 and riociguat (BAY 63-2521). Other sGC stimulators include CFM-1571, BAY 60-4552, vericiguat (BAY 1021189), the acrylamide analog A-350619 and the aminopyrimidine analogs. BAY 41-2272, BAY 41-8543 and riociguat induced marked dose-dependent redns. in mean pulmonary arterial pressure and vascular resistance with a concomitant increase in cardiac output, and they also reversed vascular remodelling and right heart hypertrophy in several exptl. models of PH. Riociguat is the first sGC stimulator that has entered clin. development. Clin. trials have shown that it significantly improves pulmonary vascular haemodynamics and increases exercise ability in patients with pulmonary arterial hypertension (PAH), chronic thromboembolic PH and PH assocd. with interstitial lung disease. Furthermore, riociguat reduces mean pulmonary arterial pressure in patients with PH assocd. with chronic obstructive pulmonary disease and improves cardiac index and pulmonary vascular resistance in patients with PH assocd. with left ventricular systolic dysfunction. These promising results suggest that sGC stimulators may constitute a valuable new therapy for PH. Other trials of riociguat are in progress, including a study of the haemodynamic effects and safety of riociguat in patients with PH assocd. with left ventricular diastolic dysfunction, and long-term extensions of the phase 3 trials investigating the efficacy and safety of riociguat in patients with PAH and chronic thromboembolic PH. Finally, sGC stimulators may also have potential therapeutic applications in other diseases, including heart failure, lung fibrosis, scleroderma and sickle cell disease.
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Ghofrani, H.-A.; Galie, N.; Grimminger, F.; Gruenig, E.; Humbert, M.; Jing, Z.-C.; Keogh, A. M.; Langleben, D.; Kilama, M. O.; Fritsch, A.; Neuser, D.; Rubin, L. J. Riociguat for the treatment of pulmonary arterial hypertension N. Engl. J. Med. 2013, 369, 330– 340 DOI: 10.1056/NEJMoa1209655
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Riociguat for the treatment of pulmonary arterial hypertension
Ghofrani, Hossein-Ardeschir; Galie, Nazzareno; Grimminger, Friedrich; Gruenig, Ekkehard; Humbert, Marc; Jing, Zhi-Cheng; Keogh, Anne M.; Langleben, David; Kilama, Michael Ochan; Fritsch, Arno; Neuser, Dieter; Rubin, Lewis J.
New England Journal of Medicine (2013), 369 (4), 330-340CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
BACKGROUND: Riociguat, a sol. guanylate cyclase stimulator, has been shown in a phase 2 trial to be beneficial in the treatment of pulmonary arterial hypertension. METHODS: In this phase 3, double-blind study, we randomly assigned 443 patients with symptomatic pulmonary arterial hypertension to receive placebo, riociguat in individually adjusted doses of up to 2.5 mg three times daily (2.5 mg-max. group), or riociguat in individually adjusted doses that were capped at 1.5 mg three times daily (1.5 mg-max. group). The 1.5 mg-max. group was included for exploratory purposes, and the data from that group were analyzed descriptively. Patients who were receiving no other treatment for pulmonary arterial hypertension and patients who were receiving endothelin-receptor antagonists or (nonintravenous) prostanoids were eligible. The primary end point was the change from baseline to the end of week 12 in the distance walked in 6 min. Secondary end points included the change in pulmonary vascular resistance, N-terminal pro-brain natriuretic peptide (NT-proBNP) levels, World Health Organization (WHO) functional class, time to clin. worsening, score on the Borg dyspnea scale, quality-of-life variables, and safety. RESULTS: By week 12, the 6-min walk distance had increased by a mean of 30 m in the 2.5 mg-max. group and had decreased by a mean of 6 m in the placebo group (least-squares mean difference, 36 m; 95% confidence interval, 20 to 52; P<0.001). Prespecified subgroup analyses showed that riociguat improved the 6-min walk distance both in patients who were receiving no other treatment for the disease and in those who were receiving endothelin-receptor antagonists or prostanoids. There were significant improvements in pulmonary vascular resistance (P<0.001), NT-proBNP levels (P<0.001), WHO functional class (P = 0.003), time to clin. worsening (P = 0.005), and Borg dyspnea score (P = 0.002). The most common serious adverse event in the placebo group and the 2.5 mg-max. group was syncope (4% and 1%, resp.). CONCLUSIONS: Riociguat significantly improved exercise capacity and secondary efficacy end points in patients with pulmonary arterial hypertension.
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Ghofrani, H.-A.; D’Armini, A. M.; Grimminger, F.; Hoeper, M. M.; Jansa, P.; Kim, N. H.; Mayer, E.; Simonneau, G.; Wilkins, M. R.; Fritsch, A.; Neuser, D.; Weimann, G.; Wang, C. Riociguat for the treatment of chronic thromboembolic pulmonary hypertension N. Engl. J. Med. 2013, 369, 319– 329 DOI: 10.1056/NEJMoa1209657
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Riociguat for the treatment of chronic thromboembolic pulmonary hypertension
Ghofrani, Hossein-Ardeschir; D'Armini, Andrea M.; Grimminger, Friedrich; Hoeper, Marius M.; Jansa, Pavel; Kim, Nick H.; Mayer, Eckhard; Simonneau, Gerald; Wilkins, Martin R.; Fritsch, Arno; Neuser, Dieter; Weimann, Gerrit; Wang, Chen
New England Journal of Medicine (2013), 369 (4), 319-329CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
BACKGROUND Riociguat, a member of a new class of compds. (sol. guanylate cyclase stimulators), has been shown in previous clin. studies to be beneficial in the treatment of chronic thromboembolic pulmonary hypertension. METHODS In this phase 3, multicenter, randomized, double-blind, placebo-controlled study, we randomly assigned 261 patients with inoperable chronic thromboembolic pulmonary hypertension or persistent or recurrent pulmonary hypertension after pulmonary endarterectomy to receive placebo or riociguat. The primary end point was the change from baseline to the end of week 16 in the distance walked in 6 min. Secondary end points included changes from baseline in pulmonary vascular resistance, N-terminal pro-brain natriuretic peptide (NT-proBNP) level, World Health Organization (WHO) functional class, time to clin. worsening, Borg dyspnea score, quality-of-life variables, and safety. RESULTS By week 16, the 6-min walk distance had increased by a mean of 39 m in the riociguat group, as compared with a mean decrease of 6 m in the placebo group (least-squares mean difference, 46 m; 95% confidence interval [CI], 25 to 67; P<0.001). Pulmonary vascular resistance decreased by 226 dyn·sec·cm-5 in the riociguat group and increased by 23 dyn·sec·cm-5 in the placebo group (least-squares mean difference, -246 dyn·sec·cm-5; 95% CI, -303 to -190; P<0.001). Riociguat was also assocd. with significant improvements in the NT-proBNP level (P<0.001) and WHO functional class (P = 0.003). The most common serious adverse events were right ventricular failure (in 3% of patients in each group) and syncope (in 2% of the riociguat group and in 3% of the placebo group). CONCLUSIONS Riociguat significantly improved exercise capacity and pulmonary vascular resistance in patients with chronic thromboembolic pulmonary hypertension.
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Hambly, N.; Granton, J. Riociguat for the treatment of pulmonary hypertension Expert Rev. Respir. Med. 2015, 9, 679– 695 DOI: 10.1586/17476348.2015.1106316
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Riociguat for the treatment of pulmonary hypertension
Hambly, Nathan; Granton, John
Expert Review of Respiratory Medicine (2015), 9 (6), 679-695CODEN: ERRMBF; ISSN:1747-6348. (Taylor & Francis Ltd.)
A review. Nitric oxide (NO) is a crit. signaling mol. in the pulmonary vasculature. NO activates sol. guanylate cyclase (sGC) resulting in the synthesis of cGMP - a key mediator of pulmonary artery vasodilatation that may also inhibit smooth muscle proliferation and platelet aggregation. Pulmonary hypertension, a serious, progressive and often fatal disease is characterized by NO-sGC-sGMP pathway dysregulation. Riociguat is a member of a novel therapeutic class known as sol. guanylate stimulators. Riociguat has a dual mode of action, acting in synergy with endogenous NO and also directly stimulating sGC independently of NO availability. Phase 3 randomized control trials have demonstrated that riociguat improves clin., physiol. and hemodynamic parameters in patients with pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. In this review we will discuss the pharmacol. properties of riociguat and its appropriate implementation into clin. practice.
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Frey, R.; Mueck, W.; Unger, S.; Artmeier-Brandt, U.; Weimann, G.; Wensing, G. Single-dose pharmacokinetics, pharmacodynamics, tolerability, and safety of the soluble guanylate cyclase stimulator BAY 63–2521: an ascending-dose study in healthy male volunteers J. Clin. Pharmacol. 2008, 48, 926– 934 DOI: 10.1177/0091270008319793
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Single-dose pharmacokinetics, pharmacodynamics, tolerability, and safety of the soluble guanylate cyclase stimulator BAY 63-2521: an ascending-dose study in healthy male volunteers
Frey, Reiner; Mueck, Wolfgang; Unger, Sigrun; Artmeier-Brandt, Ulrike; Weimann, Gerrit; Wensing, Georg
Journal of Clinical Pharmacology (2008), 48 (8), 926-934CODEN: JCPCBR; ISSN:0091-2700. (Sage Publications)
The aim of the study was to assess the safety, tolerability, pharmacokinetics, and pharmacodynamics of BAY 63-2521, a new drug in development for pulmonary hypertension. Fifty-eight healthy male volunteers received a single oral dose of BAY 63-2521 (0.25-5 mg) or placebo. No serious adverse events were reported; there were no life-threatening events. Heart rate over 1 min, an indicator of the effect of a vasodilating agent on the cardiovascular system in healthy subjects, was increased dose dependently vs. placebo at BAY 63-2521 doses of 1 to 5 mg (P < .01). Mean arterial and diastolic pressures were decreased vs. placebo at doses of 1 mg (P < .05) and 5 mg (P < .01). Systolic pressure was not significantly affected. BAY 63-2521 was readily absorbed and exhibited dose-proportional pharmacokinetics. The pharmacodynamic and pharmacokinetic properties of BAY 63-2521 suggest that it can offer a unique mode of action in the treatment of pulmonary hypertension.
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Gheorghiade, M.; Greene, S. J.; Butler, J.; Filippatos, G.; Lam, C. S.; Maggioni, A. P.; Ponikowski, P.; Shah, S. J.; Solomon, S. D.; Kraigher-Krainer, E.; Samano, E. T.; Müller, K.; Roessig, L.; Pieske, B. Effect of vericiguat, a soluble guanylate cyclase stimulator, on natriuretic peptide levels in patients with worsening chronic heart failure and reduced ejection fraction: the SOCRATES-REDUCED randomized trial JAMA, J. Am. Med. Assoc. 2015, 314, 2251– 2262 DOI: 10.1001/jama.2015.15734
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Pieske, B.; Butler, J.; Filippatos, G.; Lam, C.; Maggioni, A. P.; Ponikowski, P.; Shah, S.; Solomon, S.; Kraigher-Krainer, E.; Samano, E. T.; Scalise, A. V.; Mueller, K.; Roessig, L.; Gheorghiade, M. Rationale and design of the SOluble guanylate Cyclase stimulatoR in heArT failurE Studies (SOCRATES) Eur. J. Heart Failure 2014, 16, 1026– 38 DOI: 10.1002/ejhf.135
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Rationale and design of the SOluble guanylate Cyclase stimulatoR in heArT failurE Studies (SOCRATES)
Pieske, Burkert; Butler, Javed; Filippatos, Gerasimos; Lam, Carolyn; Maggioni, Aldo Pietro; Ponikowski, Piotr; Shah, Sanjiv; Solomon, Scott; Kraigher-Krainer, Elisabeth; Samano, Eliana Tibana; Scalise, Andrea Viviana; Mueller, Katharina; Roessig, Lothar; Gheorghiade, Mihai; N/A
European Journal of Heart Failure (2014), 16 (9), 1026-1038CODEN: EJHFFS; ISSN:1388-9842. (John Wiley & Sons Ltd.)
Aims : The clin. outcomes for patients with worsening chronic heart failure (WCHF) remain exceedingly poor despite contemporary evidence-based therapies, and effective therapies are urgently needed. Accumulating evidence supports augmentation of cyclic guanosine monophosphate (cGMP) signalling as a potential therapeutic strategy for HF with reduced or preserved ejection fraction (HFrEF and HFpEF, resp.). Direct sol. guanylate cyclase (sGC) stimulators target reduced cGMP generation due to insufficient sGC stimulation and represent a promising method for cGMP enhancement. Methods : The phase II Sol. guanylate Cyclase stimulatoR in heArT failurE Study (SOCRATES) program consists of two randomized, parallel-group, placebo-controlled, double-blind, multicentre studies, SOCRATES-REDUCED (in patients with LVEF <45%) and SOCRATES-PRESERVED (in those with LVEF ≥45%), that will explore the pharmacodynamic effects, safety and tolerability, and pharmacokinetics of four dose regimens of the once-daily oral sGC stimulator vericiguat (BAY 1021189) over 12 wk compared with placebo. These studies will enrol patients stabilized during hospitalization for HF at the time of discharge or within 4 wk thereafter. The primary endpoint in SOCRATES-REDUCED is change in NT-proBNP at 12 wk. The primary endpoints in SOCRATES-PRESERVED are change in NT-proBNP and left atrial vol. at 12 wk. Perspectives : SOCRATES will be the first program to enrol specifically both inpatients and outpatients with WCHF and patients with reduced or preserved ejection fraction. Results will inform the benefits of pursuing subsequent event-driven clin. outcome trials with sGC stimulators in this patient population. Trial registration : NCT01951625 (SOCRATES-REDUCED) and NCT01951638 (SOCRATES-PRESERVED).
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Clinicaltrials.gov. Identifier: NCT02861534 (accessed August 5, 2016).
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Mittendorf, J.; Weigand, S.; Alonso-Alija, C.; Bischoff, E.; Feurer, A.; Gerisch, M.; Kern, A.; Knorr, A.; Lang, D.; Muenter, K.; Radtke, M.; Schirok, H.; Schlemmer, K.-H.; Stahl, E.; Straub, A.; Wunder, F.; Stasch, J.-P. Discovery of riociguat (BAY 63–2521): a potent, oral stimulator of soluble guanylate cyclase for the treatment of pulmonary hypertension ChemMedChem 2009, 4, 853– 865 DOI: 10.1002/cmdc.200900014
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Discovery of Riociguat (BAY 63-2521): A Potent, Oral Stimulator of Soluble Guanylate Cyclase for the Treatment of Pulmonary Hypertension
Mittendorf, Joachim; Weigand, Stefan; Alonso-Alija, Cristina; Bischoff, Erwin; Feurer, Achim; Gerisch, Michael; Kern, Armin; Knorr, Andreas; Lang, Dieter; Muenter, Klaus; Radtke, Martin; Schirok, Hartmut; Schlemmer, Karl-Heinz; Stahl, Elke; Straub, Alexander; Wunder, Frank; Stasch, Johannes-Peter
ChemMedChem (2009), 4 (5), 853-865CODEN: CHEMGX; ISSN:1860-7179. (Wiley-VCH Verlag GmbH & Co. KGaA)
Direct stimulation of sol. guanylate cyclase (sGC),a key signal-transduction enzyme activated by nitric oxide (NO), represents a promising therapeutic strategy for the treatment of a range of diseases, including the severely disabling pulmonary hypertension (PH). Impairments of the NO-sGC signaling pathway have been implicated in the pathogenesis of cardiovascular and other diseases. Optimization of the unfavorable drug metab. and pharmacokinetic (DMPK) profile of previous sGC stimulators provided riociguat I, which is currently being investigated in phase III clin. trials for the oral treatment of PH. The compd. I exhibited an improved DMPK profile and exerted strong effects on pulmonary hemodynamics and exercise capacity in patients with PH.
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Gnoth, M. J.; Hopfe, P. M.; Czembor, W. Determination of riociguat and its major human metabolite M-1 in human plasma by stable-isotope dilution LCMS/MS Bioanalysis 2015, 7, 193– 205 DOI: 10.4155/bio.14.257
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Becker, C.; Frey, R.; Thomas, D.; Reber, M.; Weimann, G.; Arens, E. R.; Mück, W.; Unger, S.; Dietrich, H. Pharmacokinetic interaction of riociguat with ketoconazole, clarithromycin, and midazolam Pulm. Circ. 2016, 6, S49– 57 DOI: 10.1086/685016
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Pharmacokinetic interaction of riociguat with ketoconazole, clarithromycin, and midazolam
Becker, Corina; Frey, Reiner; Unger, Sigrun; Thomas, Dirk; Reber, Michael; Weimann, Gerrit; Dietrich, Hartmut; Arens, Erich R.; Mueck, Wolfgang
Pulmonary Circulation (2016), 6 (Suppl.1), S49-S57CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
Riociguat is a sol. guanylate cyclase stimulator for the treatment of pulmonary hypertension that is principally metabolized via the cytochrome P 450 (CYP) pathway. Three studies in healthy males investigated potential pharmacokinetic interactions between riociguat and CYP inhibitors (ketoconazole, clarithromycin, and midazolam). In two studies, subjects were pretreated with either oncedaily ketoconazole 400 mg or twice-daily clarithromycin 500 mg for 4 days before cotreatment with either riociguat 0.5 mg ± ketoconazole 400 mg or riociguat 1.0 mg ± clarithromycin 500 mg. In the third study, subjects received riociguat 2.5 mg 3 times daily (tid) for 3 days, followed by cotreatment with riociguat 2.5 mg tid ± midazolam 7.5 mg. Pharmacokinetic parameters, the effect of smoking on riociguat pharmacokinetics, safety, and tolerability were assessed. Pre- and cotreatment with ketoconazole and clarithromycin led to increased riociguat exposure. Pre- and cotreatment with riociguat had no significant effect on midazolam plasma concns. In all studies, the bioavailability of riociguat was reduced in smokers because its clearance to the metabolite M1 increased. Riociguat ± ketoconazole, clarithromycin, or midazolam was generally well tolerated. The most common treatment-emergent adverse events (TEAEs) across all studies were headache and dyspepsia. One serious TEAE was reported in the midazolam study. Owing to the potential for hypotension, concomitant use of riociguat with multipathway inhibitors, such as ketoconazole, should be approached with caution. Coadministration of riociguat with strong CYP3A4 inhibitors, for example, clarithromycin, does not require addnl. dose adjustment. No significant drugdrug interaction was revealed between riociguat and midazolam.
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Frey, R.; Becker, C.; Unger, S.; Schmidt, A.; Wensing, G.; Mück, W. Assessment of the effects of renal impairment and smoking on the pharmacokinetics of a single oral dose of the soluble guanylate cyclase stimulator riociguat (BAY 63–2521) Pulm. Circ. 2016, 6, S15– 26 DOI: 10.1086/685017
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Assessment of the effects of renal impairment and smoking on the pharmacokinetics of a single oral dose of the soluble guanylate cyclase stimulator riociguat (BAY 63-2521)
Frey, Reiner; Becker, Corina; Unger, Sigrun; Schmidt, Anja; Wensing, Georg; Mueck, Wolfgang
Pulmonary Circulation (2016), 6 (Suppl.1), S15-S26CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
Renal impairment is a common comborbidity in patients with pulmonary hypertension. The breakdown of riociguat, an oral sol. guanylate cyclase stimulator used to treat pulmonary hypertension, may be affected by smoking because polycyclic arom. hydrocarbons in tobacco smoke induce expression of one of the metabolizing enzymes, CYP1A1. Two nonrandomized, nonblinded studies were therefore performed to investigate the pharmacokinetics and safety of a single oral dose of riociguat 1.0 mg in individuals with mild, moderate, or severe renal impairment compared with age-, wt.-, and sex-matched healthy controls, including either smokers and nonsmokers (study I) or nonsmokers alone (study II). Pharmacokinetic analyses focused on the integrated per-protocol data set of both studies (N = 63). In patients with renal impairment, the renal clearance of riociguat was reduced and its terminal half-life prolonged compared with those in healthy controls. There was a monotonic relationship between creatinine clearance on treatment day and riociguat renal clearance (R2 = 0.62). However, increased riociguat exposure with decreasing renal function was not strictly proportional. Riociguat exposure appeared to be greater in nonsmokers than in the combined population of smokers and nonsmokers, irresp. of renal function. Adverse events were mild to moderate and in line with the mode of action of riociguat. No serious adverse events occurred. In conclusion, renal impairment was assocd. with reduced riociguat clearance compared with that in controls; however, riociguat exposure in patients with renal impairment was highly variable, and ranges overlapped with those obsd. in healthy controls.
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Saleh, S.; Frey, R.; Becker, C.; Unger, S.; Wensing, G.; Mück, W. Bioavailability, pharmacokinetics, and safety of riociguat given as an oral suspension or crushed tablet with and without food Pulm. Circ. 2016, 6, S66– 74 DOI: 10.1086/685020
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Bioavailability, pharmacokinetics, and safety of riociguat given as an oral suspension or crushed tablet with and without food
Saleh, Soundos; Frey, Reiner; Becker, Corina; Unger, Sigrun; Wensing, Georg; Mueck, Wolfgang
Pulmonary Circulation (2016), 6 (Suppl.1), S66-S74CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
Riociguat is approved for the treatment of pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. Some patients have difficulty swallowing tablets; therefore, 2 randomized, nonblinded, crossover studies compared the relative bioavailability of riociguat oral suspensions and immediate-release (IR) tablet and of crushed-tablet prepns. vs. whole IR tablet. In study 1, 30 healthy subjects received 5 single riociguat doses: 0.3 and 2.4 mg (0.15 mg/mL suspensions), 0.15 mg (0.03 mg/mL), and 1.0 mg (whole IR tablet) under fasted conditions and 2.4 mg (0.15 mg/mL) after a high-fat, high-calorie American-style breakfast. In study 2, 25 healthy men received 4 single 2.5-mg doses: whole IR tablet and crushed IR tablet suspended in applesauce and water, resp., under fasted conditions, and whole IR tablet after a continental breakfast. In study 1, dose-normalized pharmacokinetics of riociguat oral suspensions and 1.0-mg whole IR tablet were similar in fasted conditions; 90% confidence intervals for riociguat area under the curve (AUC) to dose and mean max. concn. (Cmax) to dose were within bioequivalence criteria. After food, dosenormalized AUC and Cmax decreased by 15% and 38%, resp. In study 2, riociguat exposure was similar for all prepns.; AUC ratios for crushed-IR-tablet prepns. to whole IR tablet were within bioequivalence criteria. The Cmax increased by 17% for crushed IR tablet in water vs. whole IR tablet. Food intake decreased Cmax of the whole tablet by 16%, with unaltered AUC vs. fasted conditions. Riociguat bioavailability was similar between the oral suspensions and the whole IR tablet; exposure was similar between whole IR tablet and crushed-IR-tablet prepns. Minor food effects were obsd. Results suggest that riociguat formulations are interchangeable.
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Zhao, X.; Wang, Z.; Wang, Y.; Zhang, H.; Blode, H.; Yoshikawa, K.; Becker, C.; Unger, S.; Frey, R.; Cui, Y. Pharmacokinetics of the soluble guanylate cyclase stimulator riociguat in healthy young Chinese male non-smokers and smokers: results of a randomized, double-blind, placebo-controlled study Clin. Pharmacokinet. 2016, 55, 615– 624 DOI: 10.1007/s40262-015-0337-4
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Pharmacokinetics of the Soluble Guanylate Cyclase Stimulator Riociguat in Healthy Young Chinese Male Non-Smokers and Smokers: Results of a Randomized, Double-Blind, Placebo-Controlled Study
Zhao, Xia; Wang, Zining; Wang, Yukun; Zhang, Hong; Blode, Hartmut; Yoshikawa, Kenichi; Becker, Corina; Unger, Sigrun; Frey, Reiner; Cui, Yimin
Clinical Pharmacokinetics (2016), 55 (5), 615-624CODEN: CPKNDH; ISSN:0312-5963. (Springer International Publishing AG)
Background and Objectives: The aim of this study was to investigate the pharmacokinetics, safety, and tolerability of riociguat after single and multiple oral doses of 1 or 2 mg three times daily (tid), and to det. the effect of smoking on riociguat pharmacokinetics in Chinese men. Methods: In a randomized, double-blind, placebo-controlled, single-center study stratified for smokers and non-smokers, healthy Chinese men aged 18-45 years received two riociguat doses: Dose Step 1 (1 mg) then Dose Step 2 (2 mg) conducted after the safety and tolerability at Dose Step 1 was confirmed. For each step, 12 subjects received riociguat and six received placebo. A single dose was given on Day 1, followed by a 48-h pharmacokinetic profile. Multiple-dose treatment tid was then given for 6 days (Days 3-8), with a last single dose on Day 9, followed by a 72-h pharmacokinetic profile. Primary outcomes were pharmacokinetic parameters for riociguat after single and multiple dosing. Results: Thirty-six subjects (18 smokers; 18 non-smokers) were randomized and provided valid pharmacokinetic data. Riociguat and its pharmacol. active metabolite M1 (BAY 60-4552) showed nearly dose-proportional pharmacokinetics. Accumulation was minimal in smokers and approx. two-fold in non-smokers. Exposure for riociguat was decreased by ≥60 % in smokers. No serious or significant adverse events occurred during the study. Conclusions: Riociguat pharmacokinetics showed dose proportionality in healthy Chinese men, as previously demonstrated in healthy white male individuals. Exposure to riociguat was substantially decreased in smokers compared with non-smokers. Riociguat was well tolerated in Chinese men.
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Highlights of Prescribing Information: Riociguat (Adempas); U.S. Food and Drug Administration: Silver Spring, MD, 2014; http://www.accessdata.fda.gov/drugsatfda_docs/label/2014/204819s002lbl.pdf (accessed April 4, 2014).
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Bitterli, P.; Erlenmeyer, H. Some derivatives of triazolopyrimidine Helv. Chim. Acta 1951, 34, 835– 840 DOI: 10.1002/hlca.19510340311
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Reinecke, M. G.; Woodrow, T. A.; Brown, E. S. Pyrazolo[3,4-c]pyridazines from hydrazine and aminothiophenecarboxylates J. Org. Chem. 1992, 57, 1018– 1021 DOI: 10.1021/jo00029a046
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Wunder, F.; Stasch, J.-P.; Hütter, J.; Alonso-Alija, C.; Hüser, J.; Lohrmann, E. A cell-based cGMP assay useful for ultra-high-throughput screening and identification of modulators of the nitric oxide/cGMP pathway Anal. Biochem. 2005, 339, 104– 112 DOI: 10.1016/j.ab.2004.12.025
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A cell-based cGMP assay useful for ultra-high-throughput screening and identification of modulators of the nitric oxide/cGMP pathway
Wunder, Frank; Stasch, Johannes-Peter; Huetter, Joachim; Alonso-Alija, Cristina; Hueser, Joerg; Lohrmann, Emanuel
Analytical Biochemistry (2005), 339 (1), 104-112CODEN: ANBCA2; ISSN:0003-2697. (Elsevier)
We have established a rapid, homogeneous, cell-based, and highly sensitive assay for guanosine 3'-5'-cyclic monophosphate (cGMP) that is suitable for fully automated ultra-high-throughput screening. In this assay system, cGMP prodn. is monitored in living cells via Ca2+ influx through the olfactory cyclic nucleotide-gated cation channel CNGA2, acting as the intracellular cGMP sensor. A stably transfected Chinese hamster ovary (CHO) cell line was generated recombinantly expressing sol. guanylate cyclase, CNGA2, and aequorin as a luminescence indicator for the intracellular calcium concn. This cell line was used to screen more than 900,000 compds. in an automated ultra-high-throughput screening assay using 1536-well microtiter plates. In this way, we have been able to identify BAY 58-2667, a member of a new class of amino dicarboxylic acids that directly activate sol. guanylate cyclase. The assay system allows the real-time cGMP detection within living cells and makes it possible to screen for activators and inhibitors of enzymes involved in the nitric oxide/cGMP pathway.
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Hillisch, A.; Heinrich, N.; Wild, H. Computational chemistry in the pharmaceutical industry: from childhood to adolescence ChemMedChem 2015, 10, 1958– 1962 DOI: 10.1002/cmdc.201500346
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Computational Chemistry in the Pharmaceutical Industry: From Childhood to Adolescence
Hillisch, Alexander; Heinrich, Nikolaus; Wild, Hanno
ChemMedChem (2015), 10 (12), 1958-1962CODEN: CHEMGX; ISSN:1860-7179. (Wiley-VCH Verlag GmbH & Co. KGaA)
Computational chem. within the pharmaceutical industry has grown into a field that proactively contributes to many aspects of drug design, including target selection and lead identification and optimization. While methodol. advancements have been key to this development, organizational developments have been crucial to our success as well. In particular, the interaction between computational and medicinal chem. and the integration of computational chem. into the entire drug discovery process have been invaluable. Over the past ten years we have shaped and developed a highly efficient computational chem. group for small-mol. drug discovery at Bayer HealthCare that has significantly impacted the clin. development pipeline. In this article we describe the setup and tasks of the computational group and discuss external collaborations. We explain what we have found to be the most valuable and productive methods and discuss future directions for computational chem. method development. We share this information with the hope of igniting interesting discussions around this topic.
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Burkhard, J. A.; Wuitschik, G.; Rogers-Evans, M.; Müller, K.; Carreira, E. M. Oxetanes as versatile elements in drug discovery and synthesis Angew. Chem., Int. Ed. 2010, 49, 9052– 9067 DOI: 10.1002/anie.200907155
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Oxetanes as Versatile Elements in Drug Discovery and Synthesis
Burkhard, Johannes A.; Wuitschik, Georg; Rogers-Evans, Mark; Mueller, Klaus; Carreira, Erick M.
Angewandte Chemie, International Edition (2010), 49 (48), 9052-9067CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
A review. Sizable resources, both financial and human, are invested each year in the development of new pharmaceutical agents. However, despite improved techniques, the new compds. often encounter difficulties in satisfying and overcoming the numerous physicochem. and many pharmacol. constraints and hurdles. Oxetanes have been shown to improve key properties when grafted onto mol. scaffolds. Of particular interest are oxetanes that are substituted only in the 3-position, since such units remain achiral and their introduction into a mol. scaffold does not create a new stereocenter. This Minireview gives an overview of the recent advances made in the prepn. and use of 3-substituted oxetanes. It also includes a discussion of the site-dependent modifications of various physicochem. and biochem. properties that result from the incorporation of the oxetane unit in mol. architectures.
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Wuitschik, G.; Carreira, E. M.; Wagner, B.; Fischer, H.; Parrilla, I.; Schuler, F.; Rogers-Evans, M.; Müller, K. Oxetanes in drug discovery: structural and synthetic insights J. Med. Chem. 2010, 53, 3227– 3246 DOI: 10.1021/jm9018788
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Oxetanes in Drug Discovery: Structural and Synthetic Insights
Wuitschik, Georg; Carreira, Erick M.; Wagner, Bjorn; Fischer, Holger; Parrilla, Isabelle; Schuler, Franz; Rogers-Evans, Mark; Muller, Klaus
Journal of Medicinal Chemistry (2010), 53 (8), 3227-3246CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)
The use of oxetanes as replacements for gem-di-Me or carbonyl groups and their effects on the aq. soly., lipophilicity, metabolic stability, and conformation for various compds. are studied; methods for the prepn. of a variety of substituted oxetanes are given. The magnitude of changes in properties and in metabolic stability with oxetane substitution depends on the structural context; for example, substitution of a gem-di-Me group with an oxetane, aq. soly. may increase by a factor of 4 to more than 4000 while reducing the rate of metabolic degrdn. in most cases. Incorporation of an oxetane into an aliph. chain increases in some cases the preference for synclinal conformations rather than antiplanar conformations of the chain. Spirocyclic oxetanes such as an oxazaspiroheptane resemble commonly used fragments in drug discovery, such as morpholines, and in some cases increase aq. soly. more effectively than morpholines. An improved chemoselective oxidn. of 3-oxetanol to 3-oxetanone is disclosed; olefination of 3-oxetanone by a variety of methods yields alkylideneoxetanes I [R = (EtO)2P(:O), OHC, O2N, EtO2C, NC, PhO2S, MeCO, 1-(4-chlorophenyl)-1-cyclobutanecarbonyl]. I (R = EtO2C, OHC, O2N) undergo addn. reactions with nucleophiles such as amines, carbonyl compds., and arylboronic acids to give oxetanes such as II. The crystal structures of a variety of oxetanes are detd.
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Leeson, P. D.; Young, R. J. Molecular property design: does everyone get it? ACS Med. Chem. Lett. 2015, 6, 722– 725 DOI: 10.1021/acsmedchemlett.5b00157
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Molecular Property Design: Does Everyone Get It?
Leeson, Paul D.; Young, Robert J.
ACS Medicinal Chemistry Letters (2015), 6 (7), 722-725CODEN: AMCLCT; ISSN:1948-5875. (American Chemical Society)
A review. The principles of mol. property optimization in drug design have been understood for decades, yet much drug discovery activity today is conducted at the periphery of historical druglike property space. Lead optimization trajectories aimed at reducing physicochem. risk, assisted by ligand efficiency metrics, could help to reduce clin. attrition rates.
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Sharkovska, Y.; Kalk, P.; Lawrenz, B.; Godes, M.; Hoffmann, L. S.; Wellkisch, K.; Geschka, S.; Relle, K.; Hocher, B.; Stasch, J.-P. Nitric oxide-independent stimulation of soluble guanylate cyclase reduces organ damage in experimental low-renin and high-renin models J. Hypertens. 2010, 28, 1666– 1675 DOI: 10.1097/HJH.0b013e32833b558c
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Nitric oxide-independent stimulation of soluble guanylate cyclase reduces organ damage in experimental low-renin and high-renin models
Sharkovska, Yuliya; Kalk, Philipp; Lawrenz, Bettina; Godes, Michael; Hoffmann, Linda Sarah; Wellkisch, Kathrin; Geschka, Sandra; Relle, Katharina; Hocher, Berthold; Stasch, Johannes-Peter
Journal of Hypertension (2010), 28 (8), 1666-1675CODEN: JOHYD3; ISSN:0263-6352. (Lippincott Williams & Wilkins)
The nitric oxide-sol. guanylate cyclase (sGC)-cGMP signal transduction pathway is impaired in different cardiovascular diseases, including pulmonary hypertension, heart failure and arterial hypertension. Riociguat is a novel stimulator of sol. guanylate cyclase (sGC). However, little is known about the effects of sGC stimulators in exptl. models of hypertension. We thus investigated the cardio-renal protective effects of riociguat in low-renin and high-renin rat models of hypertension. The vasorelaxant effect of riociguat was tested in vitro on isolated saphenous artery rings of normal and nitrate tolerant rabbits. The cardiovascular in-vivo effects of sGC stimulation were evaluated in hypertensive renin-transgenic rats treated with the nitric oxide-synthase inhibitor N-nitro-L-arginine Me ester (L-NAME) (high-renin model) and in rats with 5/6 nephrectomy (low-renin model). In both animal models, riociguat treatment improved survival and normalized blood pressure. Moreover, in the L-NAME study part, riociguat reduced cardiac target organ damage as indicated by lower plasma ANP, lower relative left ventricular wt. and lower cardiac interstitial fibrosis, and reduced renal target organ damage as indicated by lower plasma creatinine and urea, less glomerulosclerosis and less renal interstitial fibrosis. In the 5/6 nephrectomy study part, riociguat reduced cardiac target organ damage as indicated by lower plasma ANP, lower relative left ventricular wt., lower myocyte diam. and lower arterial media/lm ratio, and reduced renal target organ damage as indicated by improved creatinine clearance and less renal interstitial fibrosis. We demonstrate for the first time that the novel sGC stimulator riociguat shows in 2 independent models of hypertension a potent protection against cardiac and renal target organ damage.
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Dubin, R. F.; Shah, S. J. Soluble guanylate cyclase stimulators: a novel treatment option for heart failure associated with cardiorenal syndromes? Curr. Heart Failure Rep. 2016, 13, 132– 139 DOI: 10.1007/s11897-016-0290-z
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Soluble Guanylate Cyclase Stimulators: a Novel Treatment Option for Heart Failure Associated with Cardiorenal Syndromes?
Dubin, Ruth F.; Shah, Sanjiv J.
Current Heart Failure Reports (2016), 13 (3), 132-139CODEN: CHFRCF; ISSN:1546-9549. (Springer)
Heart failure in the setting of chronic kidney disease (CKD) is an increasingly common scenario and carries a poor prognosis. Clinicians lack tools for primary or secondary heart failure prevention in patients with cardiorenal syndromes. In patients without CKD, angiotensin-converting enzyme inhibitors (ACE-I) or angiotensin receptor blockers (ARB) and statins mitigate cardiovascular risk in large part due to salutary effects on the endothelium. In the setting of CKD, use of these therapies is limited by adverse effects of hyperkalemia in pre-dialysis CKD (ACE-I/ARB), or potential increased risk of stroke in end-stage renal disease (statins). The sol. guanylate cyclase (sGC) stimulators are a novel class of medications that promote endothelial and myocardial function with no known risk of hyperkalemia or stroke. In this review, we discuss the evidence emerging from recent clin. trials of sGC stimulators in pulmonary hypertension and heart failure, the diseased pathways involved in cardiorenal syndromes likely to be restored by sGC stimulators, and several strategies for designing future clin. trials of cardiorenal syndromes that might shorten the timeline for discovery and approval of effective cardiovascular therapies in these high-risk patients.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Griebenow, N.; Lang, D.; Wunder, F.; Huebsch, W.; Lindner, N.; Vakalopoulos, A.; Tersteegen, A. Preparation of annelated pyrimidine derivatives useful in the treatment and prophylaxis of cardiovascular diseases. WO2013030288, 2013.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Baerfacker, L.; Weigand, S. Bicyclic aza-heterocycles as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012028647, 2012.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Schirok, H.; Jautelat, R. Substituted methyl pyrimidin-5-yl carbamates as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012010578, 2012.
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Alonso-Alija, C.; Bischoff, E.; Muenter, K.; Stasch, J.-P.; Stahl, E.; Weigand, S.; Feurer, A. Preparation of [(pyrazolopyridinyl)pyrimidinyl]carbamates stimulating soluble guanylate cyclase for treating cardiovascular diseases and/or sexual dysfunction. WO 2003095451, 2003.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Wunder, F.; Li, V. M.-J.; Mittendorf, J.; Jautelat, R. Carbamate-substituted diaminopyrimidines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012010576, 2012.
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Schirok, H.; Mittendorf, J.; Stasch, J.-P.; Wunder, F.; Stoll, F.; Schlemmer, K.-H. Azabicyclic derivatives as stimulators of guanylate cyclase, their preparation, pharmaceutical compositions, and use for the treatment of cardiovascular disorders. WO 2008031513, 2008.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Mittendorf, J.; Schlemmer, K.-H.; Jautelat, R. Substituted 6-fluoro-1H-pyrazolo[4,3-b]pyridines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012059549, 2012.
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Follmann, M.; Stasch, J.-P.; Redlich, G.; Straub, A.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J. Substituted imidazopyridazines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012152630, 2012.
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Straub, A.; Feurer, A.; Alonso-Alija, C.; Stasch, J.-P.; Perzborn, E.; Huetter, J.; Dembowsky, K.; Stahl, E. Substituted pyrazole derivatives condensed with six-membered heterocyclic rings as cardiovascular agents and their preparation. WO 2000006569, 2000.
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Walsky, R. L.; Obach, R. S. Validated assays for human cytochrome P450 activities Drug Metab. Dispos. 2004, 32, 647– 660 DOI: 10.1124/dmd.32.6.647
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Validated assays for human cytochrome P450 activities
Walsky, Robert L.; Obach, R. Scott
Drug Metabolism and Disposition (2004), 32 (6), 647-660CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)
The measurement of the effect of new chem. entities on human cytochrome P 450 marker activities using in vitro experimentation represents an important exptl. approach in drug development. In vitro drug interaction data can be used in guiding the design of clin. drug interaction studies, or, when no effect is obsd. in vitro, the data can be used in place of an in vivo study to claim that no interaction will occur in vivo. To make such a claim, it must be assured that the in vitro expts. are performed with abs. confidence in the methods used and data obtained. To meet this need, 12 semiautomated assays for human P 450 marker substrate activities have been developed and validated using approaches described in the GLP (good lab. practices) as per the code of U.S. Federal Regulations. The assays that were validated are: phenacetin O-deethylase (CYP1A2), coumarin 7-hydroxylase (CYP2A6), bupropion hydroxylase (CYP2B6), amodiaquine N-deethylase (CYP2C8), diclofenac 4'-hydroxylase and tolbutamide methylhydroxylase (CYP2C9), (S)-mephenytoin 4'-hydroxylase (CYP2C19), dextromethorphan O-demethylase (CYP2D6), chlorzoxazone 6-hydroxylase (CYP2E1), felodipine dehydrogenase, testosterone 6β-hydroxylase, and midazolam 1'-hydroxylase (CYP3A4 and CYP3A5). High-pressure liq. chromatog.-tandem mass spectrometry, using stable isotope-labeled internal stds., has been applied as the anal. method. This anal. approach, through its high sensitivity and selectivity, has permitted the use of very low incubation concns. of microsomal protein (0.01-0.2 mg/mL). Anal. assay accuracy and precision values were excellent. Enzyme kinetic and inhibition parameters obtained using these methods demonstrated high precision and were within the range of values previously reported in the scientific literature. These methods should prove useful in the routine assessments of the potential for new drug candidates to elicit pharmacokinetic drug interactions via inhibition of cytochrome P 450 activities.
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Obach, R. S.; Walsky, R. L.; Venkatakrishnan, K. Mechanism-based inactivation of human cytochrome p450 enzymes and the prediction of drug-drug interactions Drug Metab. Dispos. 2007, 35, 246– 255 DOI: 10.1124/dmd.106.012633
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Mechanism-based inactivation of human cytochrome P450 enzymes and the prediction of drug-drug interactions
Obach, R. Scott; Walsky, Robert L.; Venkatakrishnan, Karthik
Drug Metabolism and Disposition (2007), 35 (2), 246-255CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)
The ability to use vitro inactivation kinetic parameters in scaling to in vivo drug-drug interactions (DDIs) for mechanism-based inactivators of human cytochrome P 450 (P 450) enzymes was examd. using eight human P 450-selective marker activities in pooled human liver microsomes. These data were combined with other parameters (systemic Cmax, estd. hepatic inlet Cmax, fraction unbound, in vivo P 450 enzyme degrdn. rate consts. estd. from clin. pharmacokinetic data, and fraction of the affected drug cleared by the inhibited enzyme) to predict increases in exposure to drugs, and the predictions were compared with in vivo DDIs gathered from clin. studies reported in the scientific literature. In general, the use of unbound systemic Cmax as the inactivator concn. in vivo yielded the most accurate predictions of DDI with a mean -fold error of 1.64. Abbreviated in vitro approaches to identifying mechanism-based inactivators were developed. Testing potential inactivators at a single concn. (IC25) in a 30-min preincubation with human liver microsomes in the absence and presence of NADPH followed by assessment of P 450 marker activities readily identified those compds. known to be mechanism-based inactivators and represents an approach that can be used with greater throughput. Measurement of decreases in IC50 occurring with a 30-min preincubation with liver microsomes and NADPH was also useful in identifying mechanism-based inactivators, and the IC50 measured after such a preincubation was highly correlated with the kinact/K1 ratio measured after a full characterization of inactivation. Overall, these findings support the conclusion that P 450 in vitro inactivation data are valuable in predicting clin. DDIs that can occur via this mechanism.
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Obach, R. S. Predicting clearance in humans from in vitro data Curr. Top. Med. Chem. 2011, 11, 334– 339 DOI: 10.2174/156802611794480873
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Predicting clearance in humans from in vitro data
Obach, R. Scott
Current Topics in Medicinal Chemistry (Sharjah, United Arab Emirates) (2011), 11 (4), 334-339CODEN: CTMCCL; ISSN:1568-0266. (Bentham Science Publishers Ltd.)
A review. The use of in vitro metab. in scaling to predict human clearance of new chem. entities has become a commonplace activity in the research and development of new drugs. The measurement of in vitro lability in human liver microsomes, a rich source of drug metabolizing cytochrome P 450 enzymes, has become a high throughput screen in many research organizations which is a testament to its usefulness in drug design. In this chapter, the methods used to scale in vitro intrinsic clearance data to predict in vivo clearance are described. Importantly, the numerous assumptions that are required in order to use in vitro data in this manner are laid out. These include assumptions regarding the scaling process as well as tech. aspects of the generation of the in vitro data. Finally, some other drug clearance processes that have been emerging as important are described with regard to ongoing research efforts to develop clearance prediction methods.
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Artursson, P.; Karlsson, J. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells Biochem. Biophys. Res. Commun. 1991, 175, 880– 885 DOI: 10.1016/0006-291X(91)91647-U
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Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells
Artursson, P.; Karlsson, J.
Biochemical and Biophysical Research Communications (1991), 175 (3), 880-5CODEN: BBRCA9; ISSN:0006-291X.
Monolayers of a well differentiated human intestinal epithelial cell line, Caco-2, were used as a model to study passive drug absorption across the intestinal epithelium. Absorption rate consts. (expressed as apparent permeability coeffs.) were detd. for 20 drugs and peptides with different structural properties. The permeability coeffs. ranged from approx. 5 × 10-8 to 5 × 10-5 cm/s. A good correlation was obtained between data on oral absorption in humans and the results in the Caco-2 model. Drugs that are completely absorbed in humans had permeability coeffs. >1 × 10-6 cm/s. Drugs that are absorbed to >1% but <100% had permeability coeffs. of 0.1-1.0 × 10-6 cm/s while drugs and peptides that are absorbed to <1% had permeability coeffs. of ≤1 × 10-7 cm/s. The results indicate that Caco-2 monolayers can be used as a model for studies on intestinal drug absorption.
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Hoenicka, M.; Becker, E.-M.; Apeler, H.; Sirichoke, T.; Schröder, H.; Gerzer, R.; Stasch, J.-P. Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: stimulation by YC-1, nitric oxide, and carbon monoxide J. Mol. Med. (Heidelberg, Ger.) 1999, 77, 14– 23 DOI: 10.1007/s001090050292
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Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: Stimulation by YC-1, nitric oxide, and carbon monoxide
Hoenicka, Markus; Becker, Eva-Maria; Apeler, Heiner; Sirichoke, Tim; Schroder, Henning; Gerzer, Rupert; Stasch, Johannes-Peter
Journal of Molecular Medicine (Berlin) (1999), 77 (1), 14-23CODEN: JMLME8; ISSN:0946-2716. (Springer-Verlag)
Sol. guanylyl cyclase (sGC) is the main receptor for nitric oxide, a messenger mol. with multiple clin. implications. Understanding the activation of sGC is an important step for establishing new therapeutic principles. We have now overexpressed sGC in a baculovirus/Sf9 system optimized for high protein yields to facilitate spectral and kinetic studies of the activation mechanisms of this enzyme. It was expressed in a batch fermenter using a defined mixt. of viruses encoding the α1 and β1 subunits of the rat lung enzyme. The expressed enzyme was purified from the cytosolic fraction by anion exchange chromatog., hydroxyapatite chromatog., and size exclusion chromatog. By use of this new method 2.5 L culture yielded about 1 mg of apparently homogeneous sGC with a content of about one heme per heterodimer without the need of a heme reconstitution step. The enzyme did not contain stoichiometric amts. of copper. The basal activities of the purified enzyme were 153 and 1259 nmol min-1 mg-1 in the presence of Mg2+ and Mn2+, resp. The nitric oxide releasing agent 2-(N,N-diethylamino)-diazenolate-2-oxide (DEA/NO) stimulated the enzyme 160-fold with Mg2+, whereas the NO-independent activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) induced an increase in the activity of 101-fold at a concn. of 300 μM. The combination of DEA/NO (10 μM) and YC-1 (100 μM) elicited a dose-dependent synergistic stimulation with a max. of a 792-fold increase over the basal activity in the presence of Mg2+, resulting in a specific activity of 121 μmol min-1 mg-1. The synergistic stimulation of DEA/NO and YC-1 was attenuated by the sGC inhibitor 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ) (10 μM) by 94%. In a different exptl. setup a satd. carbon monoxide soln. in the absence of ambient oxygen or NO stimulated the enzyme 15-fold in the absence and 1260-fold in the presence of YC-1 compared to an argon control. The heme spectra of the enzyme showed a shift of the Soret peak from 432 to 399 and 424 nm in the presence of DEA/NO or carbon monoxide, resp. The heme spectra were not affected by YC-1 in the absence or in the presence of DEA/NO or of carbon monoxide, which reflects the fact that YC-1 does not interact directly with the heme group of the enzyme. In summary, this study shows that our expression/purifn. procedure is suitable for producing large amts. of highly pure sGC which contains one heme per heterodimer without a reconstitution step. The activator expts. show that in a synergistic stimulation with YC-1 sGC can be activated maximally both by nitric oxide and by carbon monoxide and that YC-1 does not directly act via heme. The described method should help to facilitate the investigation of the new therapeutic principle of NO-independent guanylyl cyclase activators.
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Schermuly, R. T.; Stasch, J.-P.; Pullamsetti, S. S.; Middendorff, R.; Müller, D.; Schlüter, K.-D.; Dingendorf, A.; Hackemack, S.; Kolosionek, E.; Kaulen, C.; Dumitrascu, R.; Weissmann, N.; Mittendorf, J.; Klepetko, W.; Seeger, W.; Ghofrani, H. A.; Grimminger, F. Expression and function of soluble guanylate cyclase in pulmonary arterial hypertension Eur. Respir. J. 2008, 32, 881– 891 DOI: 10.1183/09031936.00114407
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Expression and function of soluble guanylate cyclase in pulmonary arterial hypertension
Schermuly, R. T.; Stasch, J.-P.; Pullamsetti, S. S.; Middendorff, R.; Mueller, D.; Schlueter, K.-D.; Dingendorf, A.; Hackemack, S.; Kolosionek, E.; Kaulen, C.; Dumitrascu, R.; Weissmann, N.; Mittendorf, J.; Klepetko, W.; Seeger, W.; Ghofrani, H. A.; Grimminger, F.
European Respiratory Journal (2008), 32 (4), 881-891CODEN: ERJOEI; ISSN:0903-1936. (European Respiratory Society)
Alterations of the nitric oxide receptor, sol. guanylate cyclase (sGC) may contribute to the pathophysiol. of pulmonary arterial hypertension (PAH). In the present study, the expression of sGC in explanted lung tissue of PAH patients was studied and the effects of the sGC stimulator BAY 63-2521 on enzyme activity, and hemodynamics and vascular remodelling were investigated in two independent animal models of PAH. Strong upregulation of sGC in pulmonary arterial vessels in the idiopathic PAH lungs compared with healthy donor lungs was demonstrated by immunohistochem. Upregulation of sGC was detected, similarly to humans, in the structurally remodelled smooth muscle layer in chronic hypoxic mouse lungs and lungs from monocrotaline (MCT)-injected rats. BAY 63-2521 is a novel, orally available compd. that directly stimulates sGC and sensitizes it to its physiol. stimulator, nitric oxide. Chronic treatment of hypoxic mice and MCT-injected rats, with fully established PAH, with BAY 63-2521 (10 mg·kg-1·day-1) partially reversed the PAH, the right heart hypertrophy and the structural remodelling of the lung vasculature. Upregulation of sol. guanylate cyclase in pulmonary arterial smooth muscle cells was noted in human idiopathic pulmonary arterial hypertension lungs and lungs from animal models of pulmonary arterial hypertension. Stimulation of sol. guanylate cyclase reversed right heart hypertrophy and structural lung vascular remodelling. Sol. guanylate cyclase may thus offer a new target for therapeutic intervention in pulmonary arterial hypertension.
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Geschka, S.; Kretschmer, A.; Sharkovska, Y.; Evgenov, O. V.; Lawrenz, B.; Hucke, A.; Hocher, B.; Stasch, J.-P. Soluble guanylate cyclase stimulation prevents fibrotic tissue remodeling and improves survival in salt-sensitive Dahl rats PLoS One 2011, 6, e21853 DOI: 10.1371/journal.pone.0021853
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Yunhan Jiang, Pingxian Liu, Zhiqiang Qiu, Meng Zhou, Mengdi Cheng, Tao Yang. The U.S. FDA approved cardiovascular drugs from 2011 to 2023: A medicinal chemistry perspective. European Journal of Medicinal Chemistry 2024, 275 , 116593. https://doi.org/10.1016/j.ejmech.2024.116593
Lingling Wu, Mario Rodriguez, Karim El Hachem, W. H. Wilson Tang, Chayakrit Krittanawong. Management of patients with heart failure and chronic kidney disease. Heart Failure Reviews 2024, 29 (5) , 989-1023. https://doi.org/10.1007/s10741-024-10415-9
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Yuichi Hattori, Kohshi Hattori, Kuniaki Ishii, Masanobu Kobayashi. Challenging and target-based shifting strategies for heart failure treatment: An update from the last decades. Biochemical Pharmacology 2024, 224 , 116232. https://doi.org/10.1016/j.bcp.2024.116232
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Po-Cheng Chang, Hui-Ling Lee, Hung-Ta Wo, Hao-Tien Liu, Ming-Shien Wen, Chung-Chuan Chou, . Vericiguat suppresses ventricular tachyarrhythmias inducibility in a rabbit myocardial infarction model. PLOS ONE 2024, 19 (4) , e0301970. https://doi.org/10.1371/journal.pone.0301970
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Doaa M. Mustafa, Nancy Magdy, Noha F. El Azab. Different spectrophotometric methods for simultaneous quantitation of Vericiguat and its alkaline degradation product: a comparative study with greenness profile assessment. Scientific Reports 2023, 13 (1) https://doi.org/10.1038/s41598-023-50097-1
Louise F. Dow, Alfie M. Case, Megan P. Paustian, Braeden R. Pinkerton, Princess Simeon, Paul C. Trippier. The evolution of small molecule enzyme activators. RSC Medicinal Chemistry 2023, 14 (11) , 2206-2230. https://doi.org/10.1039/D3MD00399J
Siju Bi, Wenshuang Diao, Ting Zhou, Kuaile Lin, Weicheng Zhou. Development of a new synthetic route of the key intermediate of riociguat. Synthetic Communications 2023, 53 (21) , 1844-1853. https://doi.org/10.1080/00397911.2023.2252539
Frank Wunder, Johannes‐Peter Stasch, Andreas Knorr, Thomas Mondritzki, Damian Brockschnieder, Eva‐Maria Becker‐Pelster, Peter Sandner, Hanna Tinel, Gorden Redlich, Ingo V. Hartung, Alexandros Vakalopoulos, Markus Follmann. Identification and characterization of the new generation soluble guanylate cyclase stimulator BAY‐747 designed for the treatment of resistant hypertension. British Journal of Pharmacology 2023, 180 (19) , 2500-2513. https://doi.org/10.1111/bph.16142
Figueroa‐Valverde Lauro, López‐Ramos Maria, Rosas‐Nexticapa Marcela, Alvarez‐Ramirez Magdalena, Díaz‐Cedillo Francisco, Lopez‐Gutierrez Tomas. Evaluation of biological activity of some pyridine derivatives on perfusion pressure and their interaction with the M 2 muscarinic receptor. Vietnam Journal of Chemistry 2023, 61 (5) , 594-604. https://doi.org/10.1002/vjch.202300005
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Chiara Mozzini, Mauro Pagani. The Heart Failure Knights. Current Problems in Cardiology 2023, 48 (9) , 101834. https://doi.org/10.1016/j.cpcardiol.2023.101834
Aimi Yokoi, Toru Kawada, Shohei Yokota, Midori Kakuuchi, Hiroki Matsushita, Akitsugu Nishiura, Meihua Li, Kazunori Uemura, Joe Alexander, Ryou Tanaka, Keita Saku, . Impact of vericiguat on baroreflex-mediated sympathetic circulatory regulation: An open-loop analysis. PLOS ONE 2023, 18 (8) , e0286767. https://doi.org/10.1371/journal.pone.0286767
Andreas Friebe, Jan R. Kraehling, Michael Russwurm, Peter Sandner, Achim Schmidtko. The 10th International Conference on cGMP 2022: recent trends in cGMP research and development—meeting report. Naunyn-Schmiedeberg's Archives of Pharmacology 2023, 396 (8) , 1669-1686. https://doi.org/10.1007/s00210-023-02484-8
Girish Chandra, Durg Vijay Singh, Gopal Kumar Mahato, Samridhi Patel. Fluorine-a small magic bullet atom in the drug development: perspective to FDA approved and COVID-19 recommended drugs. Chemical Papers 2023, 77 (8) , 4085-4106. https://doi.org/10.1007/s11696-023-02804-5
Ghulam Shabir, Aamer Saeed, Wajeeha Zahid, Fatima Naseer, Zainab Riaz, Nafeesa Khalil, Muneeba, Fernando Albericio. Chemistry and Pharmacology of Fluorinated Drugs Approved by the FDA (2016–2022). Pharmaceuticals 2023, 16 (8) , 1162. https://doi.org/10.3390/ph16081162
Bao-Anh Tran, Erini S. Serag-Bolos, Joel Fernandez, Aimon C. Miranda. Vericiguat: The First Soluble Guanylate Cyclase Stimulator for Reduction of Cardiovascular Death and Heart Failure Hospitalization in Patients With Heart Failure Reduced Ejection Fraction. Journal of Pharmacy Practice 2023, 36 (4) , 905-914. https://doi.org/10.1177/08971900221087096
Melinton Miguel López Ruano. Uso de vericiguat en pacientes con falla cardiaca y fracción de eyección reducida. Revista Ciencia Multidisciplinaria CUNORI 2023, 7 (2) , 141-151. https://doi.org/10.36314/cunori.v7i2.233
Anna Gorący, Jakub Rosik, Joanna Szostak, Bartosz Szostak, Szymon Retfiński, Filip Machaj, Andrzej Pawlik. Improving mitochondrial function in preclinical models of heart failure: therapeutic targets for future clinical therapies?. Expert Opinion on Therapeutic Targets 2023, 27 (7) , 593-608. https://doi.org/10.1080/14728222.2023.2240021
Burkert Pieske, Elisabeth Pieske‐Kraigher, Carolyn S.P. Lam, Vojtěch Melenovský, Karen Sliwa, Yuri Lopatin, Juan Luis Arango, M. Cecilia Bahit, Christopher M. O'Connor, Mahesh J. Patel, Lothar Roessig, Daniel A. Morris, Martin Kropf, Cynthia M. Westerhout, Yinggan Zheng, Paul W. Armstrong, . Effect of vericiguat on left ventricular structure and function in patients with heart failure with reduced ejection fraction: The VICTORIA echocardiographic substudy. European Journal of Heart Failure 2023, 25 (7) , 1012-1021. https://doi.org/10.1002/ejhf.2836
Eric H. Mace, Melissa J. Kimlinger, Frederic T. Billings, Marcos G. Lopez. Targeting Soluble Guanylyl Cyclase during Ischemia and Reperfusion. Cells 2023, 12 (14) , 1903. https://doi.org/10.3390/cells12141903
Urjosee Sahana, Markus Wehland, Ulf Simonsen, Herbert Schulz, Daniela Grimm. A Systematic Review of the Effect of Vericiguat on Patients with Heart Failure. International Journal of Molecular Sciences 2023, 24 (14) , 11826. https://doi.org/10.3390/ijms241411826
Doaa M. Mustafa, Nancy Magdy, Noha F. El Azab. The first validated stability-indicating HPLC/DAD method for quantitation of Vericiguat in its pharmaceutical formulation; Application to degradation kinetic studies. Talanta 2023, 259 , 124498. https://doi.org/10.1016/j.talanta.2023.124498
Wei-Lin Liang, Bo Liang. Soluble Guanylate Cyclase Activators and Stimulators in Patients with Heart Failure. Current Cardiology Reports 2023, 25 (6) , 607-613. https://doi.org/10.1007/s11886-023-01884-9
Jay Patel, Negin Rassekh, Gregg C. Fonarow, Prakash Deedwania, Farooq H. Sheikh, Ali Ahmed, Phillip H. Lam. Guideline-Directed Medical Therapy for the Treatment of Heart Failure with Reduced Ejection Fraction. Drugs 2023, 83 (9) , 747-759. https://doi.org/10.1007/s40265-023-01887-4
Yu. N. Belenkov, M. V. Kozhevnikova. Soluble guanylate cyclase: restoration of the NO–sGC–cGMP signaling pathway activity. A new opportunity in the treatment of heart failure. Kardiologiia 2023, 63 (5) , 68-76. https://doi.org/10.18087/cardio.2023.5.n2422
Guofang Ma, Yuefang Pan, Chaoyi Qu, Feng Li. The efficacy of vericiguat for heart failure: A meta-analysis of randomized controlled trials. Medicine 2023, 102 (21) , e33807. https://doi.org/10.1097/MD.0000000000033807
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Carla Rizzo, Sara Amata, Ivana Pibiri, Andrea Pace, Silvestre Buscemi, Antonio Palumbo Piccionello. FDA-Approved Fluorinated Heterocyclic Drugs from 2016 to 2022. International Journal of Molecular Sciences 2023, 24 (9) , 7728. https://doi.org/10.3390/ijms24097728
Ahmed K. Siddiqi, Stephen J. Greene, Marat Fudim, Robert J Mentz, Javed Butler, Muhammad Shahzeb Khan. Vericiguat for the treatment of heart failure with reduced ejection fraction. Expert Review of Cardiovascular Therapy 2023, 21 (4) , 245-257. https://doi.org/10.1080/14779072.2023.2189101
Jianhua Ma, Sheng Guo, Huan Jiang, Bo Li. Efficacy and safety of vericiguat in heart failure: a meta-analysis. Journal of International Medical Research 2023, 51 (3) https://doi.org/10.1177/03000605231159333
Bo-ang Hu, Yu-lin Li, Hai-tao Han, Bin Lu, Xu Jia, Lu Han, Wei-xuan Ma, Ping Zhu, Zhi-hao Wang, Wei Zhang, Ming Zhong, Lei Zhang. Stimulation of soluble guanylate cyclase by vericiguat reduces skeletal muscle atrophy of mice following chemotherapy. Frontiers in Pharmacology 2023, 14 https://doi.org/10.3389/fphar.2023.1112123
Shuo Yuan, Dan-Shu Wang, Hui Liu, Sheng-Nan Zhang, Wei-Guang Yang, Meng Lv, Yu-Xue Zhou, Sai-Yang Zhang, Jian Song, Hong-Min Liu. New drug approvals for 2021: Synthesis and clinical applications. European Journal of Medicinal Chemistry 2023, 245 , 114898. https://doi.org/10.1016/j.ejmech.2022.114898
Rohit Patel, Yiling Fu, Ser Khang, Agnes M. Benardeau, Scott C. Thomson, Volker Vallon. Responses in Blood Pressure and Kidney Function to Soluble Guanylyl Cyclase Stimulation or Activation in Normal and Diabetic Rats. Nephron 2023, 147 (5) , 281-300. https://doi.org/10.1159/000526934
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Jingrui He, Ziyi Li, Gagan Dhawan, Wei Zhang, Alexander E. Sorochinsky, Greg Butler, Vadim A. Soloshonok, Jianlin Han. Fluorine-containing drugs approved by the FDA in 2021. Chinese Chemical Letters 2023, 34 (1) , 107578. https://doi.org/10.1016/j.cclet.2022.06.001
Qi Lou, Luyifei Li, Guangzhong Liu, Tiankai Li, Li Zhang, Yanxiang Zang, Chengchuang Zhan, Hong Wang, Weimin Li. Vericiguat reduces electrical and structural remodeling in a rabbit model of atrial fibrillation. Journal of Cardiovascular Pharmacology and Therapeutics 2023, 28 https://doi.org/10.1177/10742484231185252
Eamon P. Mulvaney, Fabiana Renzo, Rui Adão, Emilie Dupre, Lucia Bialesova, Viviana Salvatore, Helen M. Reid, Glória Conceição, Julien Grynblat, Aida Llucià-Valldeperas, Jean-Baptiste Michel, Carmen Brás-Silva, Charles E. Laurent, Luke S. Howard, David Montani, Marc Humbert, Anton Vonk Noordegraaf, Frédéric Perros, Pedro Mendes-Ferreira, B. Therese Kinsella. The thromboxane receptor antagonist NTP42 promotes beneficial adaptation and preserves cardiac function in experimental models of right heart overload. Frontiers in Cardiovascular Medicine 2022, 9 https://doi.org/10.3389/fcvm.2022.1063967
Nadine Haase, Sarah M. Kedziora, Dominik Nikolaus Müller, Ralf Dechend. Lösliche Guanylatzyklase(sGC)-Stimulation mit Vericiguat. Die Kardiologie 2022, 16 (6) , 466-478. https://doi.org/10.1007/s12181-022-00576-y
Barry H. Greenberg. Emerging Treatment Approaches to Improve Outcomes in Patients with Heart Failure. Cardiology Discovery 2022, 2 (4) , 231-240. https://doi.org/10.1097/CD9.0000000000000060
Guangchen Li, Yifu Cheng, Chi Han, Chun Song, Niu Huang, Yunfei Du. Pyrazole-containing pharmaceuticals: target, pharmacological activity, and their SAR studies. RSC Medicinal Chemistry 2022, 13 (11) , 1300-1321. https://doi.org/10.1039/D2MD00206J
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Xiao-Yan Jia, Yong-Ming Liu, Yong-Fei Wang, Jin-Yang An, Ke-Ling Peng, Hua Wang. Bibliometric study of soluble guanylate cyclase stimulators in cardiovascular research based on web of science from 1992 to 2021. Frontiers in Pharmacology 2022, 13 https://doi.org/10.3389/fphar.2022.963255
Masashi Tawa, Tomio Okamura. Factors influencing the soluble guanylate cyclase heme redox state in blood vessels. Vascular Pharmacology 2022, 145 , 107023. https://doi.org/10.1016/j.vph.2022.107023
Karel Urbánek. Vericiguat: pharmacological profile. Klinická farmakologie a farmacie 2022, 36 (2) , 77-80. https://doi.org/10.36290/far.2022.013
Alberto Aimo, Vincenzo Castiglione, Giuseppe Vergaro, Giorgia Panichella, Michele Senni, Carlo Mario Lombardi, Michele Emdin. The place of vericiguat in the landscape of treatment for heart failure with reduced ejection fraction. Heart Failure Reviews 2022, 27 (4) , 1165-1171. https://doi.org/10.1007/s10741-021-10146-1
Vojtěch Melenovský. (Vericiguat - soluble guanylate cyclase stimulator, in therapy of heart failure). Cor et Vasa 2022, 64 (3) , 316-319. https://doi.org/10.33678/cor.2022.034
Garyfallia I. Makrynitsa, Aikaterini I. Argyriou, Aikaterini A. Zompra, Konstantinos Salagiannis, Vassiliki Vazoura, Andreas Papapetropoulos, Stavros Topouzis, Georgios A. Spyroulias. Mapping of the sGC Stimulator BAY 41-2272 Binding Site on H-NOX Domain and Its Regulation by the Redox State of the Heme. Frontiers in Cell and Developmental Biology 2022, 10 https://doi.org/10.3389/fcell.2022.925457
Michael Boettcher, Hans‐Dirk Düngen, Frank Donath, Gerd Mikus, Nikos Werner, Petra A. Thuermann, Mahir Karakas, Nina Besche, Tanja Koch, Matthias Gurniak, Corina Becker. Vericiguat in Combination with Short‐Acting Nitroglycerin in Patients With Chronic Coronary Syndromes: The Randomized, Phase Ib, VENICE Study. Clinical Pharmacology & Therapeutics 2022, 111 (6) , 1239-1247. https://doi.org/10.1002/cpt.2574
J.R. González-Juanatey, M. Anguita-Sánchez, A. Bayes-Genís, J. Comín-Colet, A. García-Quintana, A. Recio-Mayoral, J.L. Zamorano-Gómez, J.M. Cepeda-Rodrigo, L. Manzano. Vericiguat en insuficiencia cardíaca: de la evidencia científica a la práctica clínica. Revista Clínica Española 2022, 222 (6) , 359-369. https://doi.org/10.1016/j.rce.2021.12.005
J.R. González-Juanatey, M. Anguita-Sánchez, A. Bayes-Genís, J. Comín-Colet, A. García-Quintana, A. Recio-Mayoral, J.L. Zamorano-Gómez, J.M. Cepeda-Rodrigo, L. Manzano. Vericiguat in heart failure: From scientific evidence to clinical practice. Revista Clínica Española (English Edition) 2022, 222 (6) , 359-369. https://doi.org/10.1016/j.rceng.2021.12.006
Wiebke Janssen, Thomas Schwarz, Ulf Bütehorn, Wolfram Steinke, Steffen Sandmann, Dieter Lang, Armin Kern, Frank Hucke, Michael Gerisch. Pharmacokinetics and mass balance of vericiguat in rats and dogs and distribution in rats. Xenobiotica 2022, 52 (5) , 453-462. https://doi.org/10.1080/00498254.2022.2082899
Bibhuti B. Das. Therapeutic Approaches in Heart Failure with Preserved Ejection Fraction (HFpEF) in Children: Present and Future. Pediatric Drugs 2022, 24 (3) , 235-246. https://doi.org/10.1007/s40272-022-00508-z
Juan Xia, Nan Hui, Lei Tian, Chengyuan Liang, Jie Zhang, Jifang Liu, Jun Wang, Xiaodong Ren, Xiaolin Xie, Kun Wang. Development of vericiguat: The first soluble guanylate cyclase (sGC) stimulator launched for heart failure with reduced ejection fraction (HFrEF). Biomedicine & Pharmacotherapy 2022, 149 , 112894. https://doi.org/10.1016/j.biopha.2022.112894
Nicole Campbell, Julie Kalabalik-Hoganson, Kathleen Frey. Vericiguat: A Novel Oral Soluble Guanylate Cyclase Stimulator for the Treatment of Heart Failure. Annals of Pharmacotherapy 2022, 56 (5) , 600-608. https://doi.org/10.1177/10600280211041384
Barbara Petzuch, Agnès Bénardeau, Lucas Hofmeister, Jutta Meyer, Elke Hartmann, Mira Pavkovic, Ilka Mathar, Peter Sandner, Heidrun Ellinger-Ziegelbauer. Urinary miRNA Profiles in Chronic Kidney Injury—Benefits of Extracellular Vesicle Enrichment and miRNAs as Potential Biomarkers for Renal Fibrosis, Glomerular Injury, and Endothelial Dysfunction. Toxicological Sciences 2022, 187 (1) , 35-50. https://doi.org/10.1093/toxsci/kfac028
Markus Follmann, Corina Becker, Lothar Roessig, Peter Sandner, Johannes‐Peter Stasch. Discovery and Development of the Soluble Guanylate Cyclase Stimulator Vericiguat for the Treatment of Chronic Heart Failure. 2022, 27-50. https://doi.org/10.1002/9781119627784.ch3
Molly Weisert, Jennifer A. Su, Jondavid Menteer, Robert E. Shaddy, Paul F. Kantor. Drug Treatment of Heart Failure in Children: Gaps and Opportunities. Pediatric Drugs 2022, 24 (2) , 121-136. https://doi.org/10.1007/s40272-021-00485-9
Davide Benedetto Tiz, Luana Bagnoli, Ornelio Rosati, Francesca Marini, Luca Sancineto, Claudio Santi. New Halogen-Containing Drugs Approved by FDA in 2021: An Overview on Their Syntheses and Pharmaceutical Use. Molecules 2022, 27 (5) , 1643. https://doi.org/10.3390/molecules27051643
Aušra Mongirdienė, Laurynas Skrodenis, Leila Varoneckaitė, Gerda Mierkytė, Justinas Gerulis. Reactive Oxygen Species Induced Pathways in Heart Failure Pathogenesis and Potential Therapeutic Strategies. Biomedicines 2022, 10 (3) , 602. https://doi.org/10.3390/biomedicines10030602
Eddie L Myers. Bicyclic 5–6 Systems: Other Four Heteroatoms 2:2. 2022, 500-564. https://doi.org/10.1016/B978-0-12-818655-8.00056-1
Antonio García-Quintana, Alejandro Recio-Mayoral, José María Cepeda-Rodrigo, José Luis Zamorano, José Ramón González-Juanatey. Papel del vericiguat en la etiopatogenia global de la insuficiencia cardiaca con fracción de eyección reducida. Posicionamiento actual. Revista Española de Cardiología Suplementos 2022, 22 , 8-14. https://doi.org/10.1016/S1131-3587(22)00002-4
Hayah Kassis-George, Nathan J Verlinden, Sheng Fu, Manreet Kanwar. Vericiguat in Heart Failure with a Reduced Ejection Fraction: Patient Selection and Special Considerations. Therapeutics and Clinical Risk Management 2022, Volume 18 , 315-322. https://doi.org/10.2147/TCRM.S357422
Barry Greenberg. Medical Management of Patients With Heart Failure and Reduced Ejection Fraction. Korean Circulation Journal 2022, 52 (3) , 173. https://doi.org/10.4070/kcj.2021.0401
Lin Luo, Xu Yang, Kai Tang, Jianli Wu, Dejin Li, Jiuju Ran, Li Zhang, Dan Wang, Dan Zhao, Min Yu, Anfang Chen, Maya Saranathan. Efficacy of three novel drugs in the treatment of heart failure: A network meta-analysis. Medicine 2022, 101 (29) , e29415. https://doi.org/10.1097/MD.0000000000029415
Hauke Ruehs, Dagmar Klein, Matthias Frei, Joachim Grevel, Rupert Austin, Corina Becker, Lothar Roessig, Burkert Pieske, Dirk Garmann, Michaela Meyer. Population Pharmacokinetics and Pharmacodynamics of Vericiguat in Patients with Heart Failure and Reduced Ejection Fraction. Clinical Pharmacokinetics 2021, 60 (11) , 1407-1421. https://doi.org/10.1007/s40262-021-01024-y
Dmitriy Yu. Vandyshev, Evgeniya A. Kosheleva, Vladimir A. Polikarchuk, Daria A. Mangusheva, Gleb L. Denisov, Khidmet S. Shikhaliev. Regioselective synthesis of novel imidazo[1,5-b]pyridazine derivatives from diaminoimidazoles and α-acylacrylonitriles. Mendeleev Communications 2021, 31 (6) , 821-823. https://doi.org/10.1016/j.mencom.2021.11.017
David J. Cordwin, Theodore J. Berei, Kristen T. Pogue. The Role of sGC Stimulators and Activators in Heart Failure With Reduced Ejection Fraction. Journal of Cardiovascular Pharmacology and Therapeutics 2021, 26 (6) , 593-600. https://doi.org/10.1177/10742484211042706
Zh. D. Kobalava, P. V. Lazarev. Nitric oxide — soluble guanylate cyclase — cyclic guanosine monophosphate signaling pathway in the pathogenesis of heart failure and search for novel therapeutic targets. Cardiovascular Therapy and Prevention 2021, 20 (6) , 3035. https://doi.org/10.15829/1728-8800-2021-3035
Jean-Sébastien Hulot, Jean-Noël Trochu, Erwan Donal, Michel Galinier, Damien Logeart, Pascal De Groote, Yves Juillière. Vericiguat for the treatment of heart failure: mechanism of action and pharmacological properties compared with other emerging therapeutic options. Expert Opinion on Pharmacotherapy 2021, 22 (14) , 1847-1855. https://doi.org/10.1080/14656566.2021.1937121
Andrew J. S. Coats, Heli Tolppanen. Drug Treatment of Heart Failure with Reduced Ejection Fraction: Defining the Role of Vericiguat. Drugs 2021, 81 (14) , 1599-1604. https://doi.org/10.1007/s40265-021-01586-y
Amr Abdin, Michael Böhm. Renal function and vericiguat in heart failure patients: light at the end of the tunnel!. European Journal of Heart Failure 2021, 23 (8) , 1322-1324. https://doi.org/10.1002/ejhf.2280
Amr Abdin, Johann Bauersachs, Norbert Frey, Ingrid Kindermann, Andreas Link, Nikolaus Marx, Mitja Lainscak, Jonathan Slawik, Christian Werner, Jan Wintrich, Michael Böhm. Timely and individualized heart failure management: need for implementation into the new guidelines. Clinical Research in Cardiology 2021, 110 (8) , 1150-1158. https://doi.org/10.1007/s00392-021-01867-2
Shalini Murali Krishnan, Johannes Nordlohne, Lisa Dietz, Alexandros Vakalopoulos, Petra Haning, Elke Hartmann, Roland Seifert, Jörg Hüser, Ilka Mathar, Peter Sandner. Assessing the Use of the sGC Stimulator BAY-747, as a Potential Treatment for Duchenne Muscular Dystrophy. International Journal of Molecular Sciences 2021, 22 (15) , 8016. https://doi.org/10.3390/ijms22158016

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Journal of Medicinal Chemistry

Cite this: J. Med. Chem. 2017, 60, 12, 5146–5161

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Abstract
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Scheme 1
Scheme 1. Synthesis of Compounds 1–18^a
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Scheme aReagents and conditions: (a) NaOMe, DMF, 110 °C; (b) H₂ (65 bar), Raney nickel, DMF; (c) R²OCOCl, pyridine, or (i) triphosgene, pyridine, (ii) R²OH, pyridine; (d) R¹X (X = halide or trichloromethanesulfonate), LiHMDS; (e) glycolaldehyde, NaBH₃CN, AcOH, MeOH, 0 °C to rt; (f) methyl chloroformate, pyridine, rt; (g) NaHMDS, THF, 0 °C to rt.
Scheme 2
Scheme 2. Synthesis of Compounds 19 and 20^a
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Scheme aReagents and conditions: (a) methyl 2-(2-fluorophenyl)acetate, LiHMDS; (b) aq NaCl solution, DMSO, microwave, 150 °C; (c) hydrazine hydrate, pyridine, reflux; (d) H₂, 10% Pd/C, Et₃N, EtOH, THF; (e) NaH, DMF, 80 °C; (f) H₂, 10% Pd/C, pyridine; (g) methyl chloroformate, pyridine.
Scheme 3
Scheme 3. Synthesis of Compound 21^a
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Scheme aReagents and conditions: (a) hydrazine hydrate; (b) POBr₃, sulfolane, 150 °C; (c) 2-fluorobenzyl bromide, Cs₂CO₃, DMF, rt; (d) CuCN, DMSO, 150 °C; (e) NaOMe, NH₄Cl, AcOH, MeOH; (f) 1b, DMF, Et₃N, 100 °C; (g) H₂ (1 bar), 10% Pd/C, DMF; (h) methyl chloroformate, pyridine, DCM.
Scheme 4
Scheme 4. Synthesis of Compound 22^a
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Scheme aReagents and conditions: (a) MeI, K₂CO₃, acetone; (b) TBAI, KOt-Bu, DCM, then aq HCl, Et₂O; (c) dioxane, 60 °C; (d) 1,1,3,3-tetramethoxypropane, aq HCl, MeOH, EtOH, reflux; (e) NaOH; (f) HATU, NH₄Cl, Hünig’s base, DMF; (g) POCl₃, 120 °C; (h) NaOMe, NH₄Cl, MeOH; (i) 1b, DMF, Et₃N, 100 °C; (j) H₂ (1 bar), 10% Pd/C, DMF, 0 °C; (k) methyl chloroformate, pyridine.
Scheme 5
Scheme 5. Synthesis of Compound 23^a
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Scheme aReagents and conditions: (a) hydrazine hydrate, Et₃N, reflux; (b) Et₃N, MeCN; (c) Br₂, AcOH, 50 °C; (d) POCl₃, DCE, reflux; (e) H₂, 5% Pd/C, Et₃N, EtOAc; (f) NBS, DCM; (g) CuCN; (h) NaOMe, NH₄Cl, AcOH; (i) 1b, DMF, Et₃N, 100 °C; (j) H₂ (1 bar), 10% Pd/C, DMF, MeOH; (k) methyl chloroformate, pyridine.
Scheme 6
Scheme 6. Synthesis of 24^a
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Scheme aReagents and conditions: (a) Tf₂O, 70 °C, then morpholine, 5 °C, then 40 °C; (b) MeSO₃Me, 135 °C, then 100 °C; (c) 45% aq NaOH, 50 °C; (d) morpholine, Et₃N, reflux; (e) MsOH, LiCl, EtOH, reflux; (f) formamide, NaOMe, MeOH, EtOH, 95–125 °C; (g) POCl₃, sulfolane, 107 °C; (h) NaOMe, NH₄Cl, MeOH, EtOH, 65 °C; (i) 1b, DMF, Et₃N, 100 °C; (j) H₂ (60 bar), 5% Pd/C, DMF, 60 °C; (k) methyl chloroformate, i-PrOH, MeOH, then Et₃N, 50 °C.
Figure 1
Figure 1. Biotransformation products of 24 in human hepatocytes.
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Figure 2
Figure 2. Effects of 24 and NO on the stimulation of highly purified sGC and blocking effects of the sGC inhibitor ODQ.
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Figure 3
Figure 3. Effects of 24 on rat heart Langendorff preparations.
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Figure 4
Figure 4. Increase in systolic blood pressure in mmHg during the course of the study with L-NAME-treated renin transgenic rats.
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Figure 5
Figure 5. Right and left ventricle weight normalized on body weight (left/middle) and plasma atrial natriuretic peptide levels [in pg/mL] at the study end, after 3 weeks of treatment.
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Figure 6
Figure 6. Effects on proteinuria at the study end after 3 weeks of treatment.
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Figure 7
Figure 7. Kaplan–Meier survival curves.
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References
This article references 56 other publications.
1. 1
  Evgenov, O. V.; Pacher, P.; Schmidt, P. M.; Hasko, G.; Schmidt, H. H. H. W.; Stasch, J.-P. NO-independent stimulators and activators of soluble guanylate cyclase: discovery and therapeutic potential Nat. Rev. Drug Discovery 2006, 5, 755– 768 DOI: 10.1038/nrd2038
  1
  NO-independent stimulators and activators of soluble guanylate cyclase: discovery and therapeutic potential
  Evgenov, Oleg V.; Pacher, Pal; Schmidt, Peter M.; Hasko, Gyoergy; Schmidt, Harald H. H. W.; Stasch, Johannes-Peter
  Nature Reviews Drug Discovery (2006), 5 (9), 755-768CODEN: NRDDAG; ISSN:1474-1776. (Nature Publishing Group)
  A review. Sol. guanylate cyclase (sGC) is a key signal-transduction enzyme activated by nitric oxide (NO). Impaired bioavailability and/or responsiveness to endogenous NO has been implicated in the pathogenesis of cardiovascular and other diseases. Current therapies that involve the use of org. nitrates and other NO donors have limitations, including non-specific interactions of NO with various biomols., lack of response and the development of tolerance following prolonged administration. Compds. that activate sGC in an NO-independent manner might therefore provide considerable therapeutic advantages. Here we review the discovery, biochem., pharmacol. and clin. potential of heme-dependent sGC stimulators (including YC-1, BAY 41-2272, BAY 41-8543, CFM-1571 and A-350619) and heme-independent sGC activators (including BAY 58-2667 and HMR-1766).
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28XptVCltrw%253D&md5=6c2a2ad7db529a7fe366bcfa0d61a6af
2. 2
  Follmann, M.; Griebenow, N.; Hahn, M. G.; Hartung, I.; Mais, F.-J.; Mittendorf, J.; Schaefer, M.; Schirok, H.; Stasch, J.-P.; Stoll, F.; Straub, A. The chemistry and biology of soluble guanylate cyclase stimulators and activators Angew. Chem., Int. Ed. 2013, 52, 9442– 9462 DOI: 10.1002/anie.201302588
  2
  The Chemistry and Biology of Soluble Guanylate Cyclase Stimulators and Activators
  Follmann, Markus; Griebenow, Nils; Hahn, Michael G.; Hartung, Ingo; Mais, Franz-Josef; Mittendorf, Joachim; Schaefer, Martina; Schirok, Hartmut; Stasch, Johannes-Peter; Stoll, Friederike; Straub, Alexander
  Angewandte Chemie, International Edition (2013), 52 (36), 9442-9462CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
  A review. The vasodilatory properties of nitric oxide (NO) have been utilized in pharmacotherapy for more than 130 years. Still today, NO-donor drugs are important in the management of cardiovascular diseases. However, inhaled NO or drugs releasing NO and org. nitrates are assocd. with noteworthy therapeutic shortcomings, including resistance to NO in some disease states, the development of tolerance during long-term treatment, and nonspecific effects, such as post-translational modification of proteins. The beneficial actions of NO are mediated by stimulation of sol. guanylate cyclase (sGC), a heme-contg. enzyme which produces the intracellular signaling mol. cGMP. Recently, two classes of compds. have been discovered that amplify the function of sGC in a NO-independent manner, the so-called sGC stimulators and sGC activators. The most advanced drug, the sGC stimulator riociguat, has successfully undergone Phase III clin. trials for different forms of pulmonary hypertension.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3sXhtlSiu77K&md5=d84536785fcea442b23179a6d7bf7246
3. 3
  Cerra, M. C.; Pellegrino, D. Cardiovascular cGMP-generating systems in physiological and pathological conditions Curr. Med. Chem. 2007, 14, 585– 599 DOI: 10.2174/092986707780059715
  3
  Cardiovascular cGMP-generating systems in physiological and pathological conditions
  Cerra, M. C.; Pellegrino, D.
  Current Medicinal Chemistry (2007), 14 (5), 585-599CODEN: CMCHE7; ISSN:0929-8673. (Bentham Science Publishers Ltd.)
  A review. The intracellular messenger cyclic GMP (cGMP) represents the key signal in several transduction pathways throughout the animal world. In the heart cGMP signaling contributes to functional interaction of different cell types. Nitric oxide (NO) and natriuretic peptides (NPs), major autocrine-paracrine cardiovascular regulators, increment intracellular cGMP through guanylate cyclases (GCs). NO and NPs interact with two GC types: cytosolic (sol.: sGC) and membrane bound [particulate: pGC (NP receptor types A and B)], resp. Depending on sub-cellular localization and regulation of the enzymes, cGMP produced by either pGC or sGC exerts different complementary effects. The two pathways are reciprocally regulated. NPs-depending pGC is modulated by NO-cGMP signaling, and the activity of NO is influenced by cellular concns. of both NO itself and NPs. This heterologous feedback regulates GCs, linking cardiovascular autocrine-paracrine activities of NPs and NO. Importance of these cGMP converging routes goes far beyond their role under normal conditions. They are of relevance esp. in disease states when tissue and circulating levels of NPs, and local NO prodn. are altered. An example is the endothelial dysfunction assocd. with deficient NO prodn. and uncoupled endothelium-myocardium communications. In this case, NPs-pGC-cGMP could supplement the reduced activity of NO-scGC-cGMP pathway. In addn., these systems regulate cell growth and apoptosis, playing a role in myocardial pathol. morpho-functional remodeling. Here we will review recent concepts on NO/NPs dependent control of heart function in vertebrates, also focusing on cGMP-activated downstream signaling and its role in health and disease conditions.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD2sXhs1yhur8%253D&md5=e908a3f3ba079740f192df00daf01396
4. 4
  Gladwin, M. T. Deconstructing endothelial dysfunction: soluble guanylyl cyclase oxidation and the NO resistance syndrome J. Clin. Invest. 2006, 116, 2330– 2332 DOI: 10.1172/JCI29807
  4
  Deconstructing endothelial dysfunction: soluble guanylyl cyclase oxidation and the NO resistance syndrome
  Gladwin, Mark T.
  Journal of Clinical Investigation (2006), 116 (9), 2330-2332CODEN: JCINAO; ISSN:0021-9738. (American Society for Clinical Investigation)
  A review. In this issue of the JCI, Stasch and colleagues suggest that a novel drug, BAY 58-2667, potently activates a pool of oxidized and heme-free sol. guanylyl cyclase (sGC; see the related article beginning on page 2552). The increased vasodilatory potency of BAY 58-2667 the authors found in a no. of animal models of endothelial dysfunction and in human blood vessels from patients with diabetes suggests that there exists a subphenotype of endothelial dysfunction characterized by receptor-level NO resistance. Diseases assocd. with NO resistance would appear to be ideally suited for therapies directed at restoring redox homeostasis, sGC activity, and NO sensitivity.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28Xps1Sgs7Y%253D&md5=9e5f9bf0c5d662275529e43902d83368
5. 5
  Hoenicka, M.; Schmid, C. Cardiovascular effects of modulators of soluble guanylyl cyclase activity Cardiovasc. Hematol. Agents Med. Chem. 2008, 6, 287– 301 DOI: 10.2174/187152508785909555
  5
  Cardiovascular effects of modulators of soluble guanylyl cyclase activity
  Hoenicka, Markus; Schmid, Christof
  Cardiovascular & Hematological Agents in Medicinal Chemistry (2008), 6 (4), 287-301CODEN: CHAAA5; ISSN:1871-5257. (Bentham Science Publishers Ltd.)
  A review. Sol. guanylyl cyclase (sGC) is one of the key enzymes of the nitric-oxide (NO)/cyclic 3',5'-guanosine monophosphate (cGMP) pathway. Located in virtually all mammalian cells, it controls the vessel tone, smooth muscle cell growth, platelet aggregation, and leukocyte adhesion. In vivo sGC activity is mainly regulated by NO which in turn is released from L-arginine by nitric oxide synthases. One of the main diseases of the cardiovascular system, endothelial dysfunction, leads to a diminished NO synthesis and thus increases vessel tone as well as the risk of thrombosis. The predominant therapeutic approach to this condition is a NO replacement therapy, as exemplified by org. nitrates, molsidomin, and other NO releasing substances. Recent advances in drug discovery provided a variety of other approaches to activate sGC, which may help to circumvent both the tolerance problem and some non-specific actions assocd. with NO donor drugs. Substances like BAY 41-2272 stimulate sGC in a heme-dependent fashion and synergize with NO, allowing to enhance the effects both of endogenous NO and of exogenous NO donors. On the other hand, heme-independent activators like BAY 58-2667 allow to activate sGC even if it is rendered unresponsive to NO due to oxidative stress or heme loss. Furthermore, a few substances have been described as specific inhibitors of sGC that allow to alleviate the effects of excess NO prodn. as seen in shock. This review discusses the cardiovascular effects of heme-dependent and heme-independent activators as well as of inhibitors of sGC.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD1cXhtlCntbjF&md5=329ce40010581e109f96a6331b8d754f
6. 6
  Priviero, F. B.; Webb, R. C. Heme-dependent and independent soluble guanylate cyclase activators and vasodilation J. Cardiovasc. Pharmacol. 2010, 56, 229– 233 DOI: 10.1097/FJC.0b013e3181eb4e75
  6
  Heme-Dependent and Independent Soluble Guanylate Cyclase Activators and Vasodilation
  Priviero, Fernanda B. M.; Webb, R. Clinton
  Journal of Cardiovascular Pharmacology (2010), 56 (3), 229-233CODEN: JCPCDT; ISSN:0160-2446. (Lippincott Williams & Wilkins)
  A review. Since the discovery of nitric oxide (NO), which is released from endothelial cells as the main mediator of vasodilation, its target, the sol. guanylyl cyclase (sGC), has become a focus of interest for the treatment of diseases assocd. with endothelial dysfunction. NO donors were developed to suppress NO deficiency; however, tolerance to org. nitrates was reported. Non-NO-based drugs targeting sGC were developed to overcome the problem of tolerance. In this review, we briefly describe the process of sGC activation by its main physiol. activator NO and the advances in the development of drugs capable of activating sGC in a NO-independent manner. sGC stimulators, as some of these drugs are called, require the integrity of the reduced heme moiety of the prosthetic group within the sGC and therefore are called heme-dependent stimulators. Other drugs are able to activate sGC independent of heme moiety and are hence called heme-independent activators. Because pathol. conditions modulate sGC and oxidize the heme moiety, the heme-independent sGC activators could potentially become drugs of choice because of their higher affinity to the oxidized enzyme. However, these drugs are still undergoing clin. trials and are not available for clin. use.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BC3cXhtFKmur%252FF&md5=6a2d08cf3f0894a5584cb988ed6ca344
7. 7
  Packer, C. S. Soluble guanylate cyclase (sGC) down-regulation by abnormal extracellular matrix proteins as a novel mechanism in vascular dysfunction: implications in metabolic syndrome Cardiovasc. Res. 2006, 69, 302– 303 DOI: 10.1016/j.cardiores.2005.12.006
  7
  Soluble guanylate cyclase (sGC) down-regulation by abnormal extracellular matrix proteins as a novel mechanism in vascular dysfunction: Implications in metabolic syndrome
  Packer, C. Subah
  Cardiovascular Research (2006), 69 (2), 302-303CODEN: CVREAU; ISSN:0008-6363. (Elsevier B.V.)
  A review. The role of endothelial NO prodn., accumulation of extracellular matrix proteins and altered activity of sol. guanylate cyclase in endothelial dysfunction and metabolic syndrome is discussed.
  >> More from SciFinder ^®
  https://chemport.cas.org/services/resolver?origin=ACS&resolution=options&coi=1%3ACAS%3A528%3ADC%252BD28Xls1Wrtg%253D%253D&md5=c08169802b8f12d26d67479e3121a02b
8. 8
  Mayer, B.; Koesling, D. cGMP signalling beyond nitric oxide Trends Pharmacol. Sci. 2001, 22, 546– 548 DOI: 10.1016/S0165-6147(00)01889-7
  8
  cGMP signalling beyond nitric oxide
  Mayer, Bernd; Koesling, Doris
  Trends in Pharmacological Sciences (2001), 22 (11), 546-548CODEN: TPHSDY; ISSN:0165-6147. (Elsevier Science Ltd.)
  A review. Many of the physiol. effects of nitric oxide are mediated by activation of sol. guanylyl cyclase, resulting in cellular cGMP accumulation. In the 1990s, the benzylindazole deriv. YC-1 was identified as a novel modulator of cGMP signaling that exerted complex actions in a NO-independent manner. A recent study describes a high-affinity YC-1 analog that decreases blood pressure in hypertensive rats and increases bleeding time, which suggests that this drug might have therapeutic potential as a vasodilator with antiplatelet activity.
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9. 9
  Nioche, P.; Berka, V.; Vipond, J.; Minton, N.; Tsai, A. L.; Raman, C. S. Femtomolar sensitivity of a NO sensor from Clostridium botulinum Science 2004, 306, 1550– 1553 DOI: 10.1126/science.1103596
  9
  Femtomolar sensitivity of a NO sensor from Clostridium botulinum
  Nioche, Pierre; Berka, Vladimir; Vipond, Julia; Minton, Nigel; Tsai, Ah-Lim; Raman, C. S.
  Science (Washington, DC, United States) (2004), 306 (5701), 1550-1553CODEN: SCIEAS; ISSN:0036-8075. (American Association for the Advancement of Science)
  Nitric oxide (NO) is extremely toxic to Clostridium botulinum, but its mol. targets are unknown. Here, the authors identify a heme protein sensor (SONO) that displays femtomolar affinity for NO. The crystal structure of the SONO heme domain reveals a previously undescribed fold and a strategically placed tyrosine residue that modulates heme-nitrosyl coordination. Furthermore, the domain architecture of a SONO ortholog cloned from Chlamydomonas reinhardtii indicates that NO signaling through cGMP arose before the origin of multicellular eukaryotes. The authors' findings have broad implications for understanding bacterial responses to NO, as well as for the activation of mammalian NO-sensitive guanylyl cyclase.
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10. 10
  Pellicena, P.; Karow, D. S.; Boon, E. M.; Marletta, M. A.; Kuriyan, J. Crystal structure of an oxygen-binding heme domain related to soluble guanylate cyclases Proc. Natl. Acad. Sci. U. S. A. 2004, 101, 12854– 12859 DOI: 10.1073/pnas.0405188101
  10
  Crystal structure of an oxygen-binding heme domain related to soluble guanylate cyclases
  Pellicena, Patricia; Karow, David S.; Boon, Elizabeth M.; Marletta, Michael A.; Kuriyan, John
  Proceedings of the National Academy of Sciences of the United States of America (2004), 101 (35), 12854-12859CODEN: PNASA6; ISSN:0027-8424. (National Academy of Sciences)
  Sol. guanylate cyclases are nitric oxide-responsive signaling proteins in which the nitric oxide sensor is a heme-binding domain of unknown structure that we have termed the heme-NO and oxygen binding (H-NOX) domain. H-NOX domains are also found in bacteria, either as isolated domains, or are fused through a membrane-spanning region to methyl-accepting chemotaxis proteins. We have detd. the crystal structure of an oxygen-binding H-NOX domain of one such signaling protein from the obligate anaerobe Thermoanaerobacter tengcongensis at 1.77-Å resoln., revealing a protein fold unrelated to known structures. Particularly striking is the structure of the protoporphyrin IX group, which is distorted from planarity to an extent not seen before in protein-bound heme groups. Comparison of the structure of the H-NOX domain in two different crystal forms suggests a mechanism whereby alteration in the degree of distortion of the heme group is coupled to changes on the mol. surface of the H-NOX domain and potentially to changes in intermol. interactions.
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11. 11
  Wedel, B.; Humbert, P.; Harteneck, C.; Foerster, J.; Malkewitz, J.; Bohme, E.; Schultz, G.; Koesling, D. Mutation of His-105 in the β₁ subunit yields a nitric oxide-insensitive form of soluble guanylyl cyclase Proc. Natl. Acad. Sci. U. S. A. 1994, 91, 2592– 2596 DOI: 10.1073/pnas.91.7.2592
  11
  Mutation of His-105 in the β1 subunit yields a nitric oxide-insensitive form of soluble guanylyl cyclase
  Wedel, Barbara; Humbert, Peter; Harteneck, Christian; Foerster, John; Malkewitz, Jurgen; Boehme, Eycke; Schultz, Gunter; Koesling, Doris
  Proceedings of the National Academy of Sciences of the United States of America (1994), 91 (7), 2592-6CODEN: PNASA6; ISSN:0027-8424.
  Sol. guanylyl cyclase [GTP pyrophosphate-lyase (cyclizing); EC 4.6.1.2] is a hemoprotein that exists as a heterodimer; the heme moiety has been proposed to bind nitric oxide, resulting in a dramatic activation of the enzyme. Mutation of six conserved His residues reduced but did not abolish nitric oxide stimulation whereas a change of His-105 to Phe in the β1 subunit yielded a heterodimer that retained basal cyclase activity but failed to respond to nitric oxide. Heme was not detected as a component of the mutant heterodimer and protophorphyrin IX failed to stimulate enzyme activity. The activity of the His mutant was almost identical to that of the wild-type enzyme in the presence of KCN, suggesting that disruption of heme binding is the principal effect of the mutation. Thus, the mutation provides a means to inhibit the nitric oxide-sensitive guanylyl cyclase signalling pathway.
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12. 12
  Zabel, U.; Hausler, C.; Weeger, M.; Schmidt, H. H. Homodimerization of soluble guanylyl cyclase subunits. Dimerization analysis using a glutathione S-transferase affinity tag J. Biol. Chem. 1999, 274, 18149– 18152 DOI: 10.1074/jbc.274.26.18149
  12
  Homodimerization of soluble guanylyl cyclase subunits. Dimerization analysis using a glutathione S-transferase affinity tag
  Zabel, Ulrike; Hausler, Christoph; Weeger, Monika; Schmidt, Harald H. H. W.
  Journal of Biological Chemistry (1999), 274 (26), 18149-18152CODEN: JBCHA3; ISSN:0021-9258. (American Society for Biochemistry and Molecular Biology)
  Sol. guanylyl cyclase (sGC) is an α/β-heterodimeric hemoprotein that, upon interaction with the intercellular messenger mol. NO, generates cGMP. Although the related family of particulate guanylyl cyclases (pGCs) forms active homodimeric complexes, it is not known whether homodimerization of sGC subunits occurs. We report here the expression in Sf9 cells of glutathione S-transferase-tagged recombinant human sGCα1 and β1 subunits, applying a novel and rapid purifn. method based on GSH-Sepharose affinity chromatog. Surprisingly, in intact Sf9 cells, both homodimeric GSTα/α and GSTβ/β complexes were formed that were catalytically inactive. Upon coexpression of the resp. complementary subunits, GSTα/β or GSTβ/α heterodimers were preferentially formed, whereas homodimers were still detectable. When subunits were mixed after expression, e.g. GSTβ and β or GSTα and β, no dimerization was obsd. In conclusion, our data suggest the previously unrecognized possibility of a physiol. equil. between homo- and heterodimeric sGC complexes.
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13. 13
  Zabel, U.; Weeger, M.; La, M.; Schmidt, H. H. W. Human soluble guanylate cyclase: functional expression and revised isoenzyme family Biochem. J. 1998, 335, 51– 57 DOI: 10.1042/bj3350051
  13
  Human soluble guanylate cyclase: functional expression and revised isoenzyme family
  Zabel, Ulrike; Weeger, Monika; La, Mylinh; Schmidt, Harald H. H. W.
  Biochemical Journal (1998), 335 (1), 51-57CODEN: BIJOAK; ISSN:0264-6021. (Portland Press Ltd.)
  Sol. guanylate cyclase (sGC), a heterodimeric (α/β) heme protein that converts GTP to the second messenger cGMP, functions as the receptor for nitric oxide (NO) and nitrovasodilator drugs. Three distinct cDNA species of each subunit (α1-α3, β1-β3) have been reported from various species. From human sources, none of these have been expressed as functionally active enzyme. Here the authors described the expression of human α/β heterodimeric sGC in Sf9 cells yielding active recombinant enzyme that was stimulated by the nitrovasodilator sodium nitroprusside or the NO-independent activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1). At the protein level, both α and β subunits were detected in human tissues, suggesting co-expression also in vivo. Moreover, resequencing of the human cDNA clones [originally termed α3 and β3] revealed several sequencing errors in human α3; correction of these eliminated major regions of divergence from rat and bovine α1. As human β3 also displays more than 98% similarity to rat and bovine β1 at the amino acid level, α3 and β3 represent the human homologs of rat and bovine α1 and β1, and the isoenzyme family is decreased to two isoforms for each subunit (α1, α2; β1, β2). Having access to the human key enzyme of NO signaling will now permit the study of novel sGC-modulating compds. with therapeutic potential.
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14. 14
  Murad, F. Shattuck Lecture. Nitric oxide and cyclic GMP in cell signaling and drug development N. Engl. J. Med. 2006, 355, 2003– 2011 DOI: 10.1056/NEJMsa063904
  14
  Nitric oxide and cyclic GMP in cell signaling and drug development
  Murad, Ferid
  New England Journal of Medicine (2006), 355 (19), 2003-2011CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
  A review. The topics discussed include: guanylyl cyclase activity; endothelial activity; nitric oxide synthase; activation of sol. guanylyl cyclase; steady-state levels of cGMP; and cGMP signaling in drug development.
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15. 15
  Stasch, J. P.; Hobbs, A. J. NO-independent, haem-dependent soluble guanylate cyclase stimulators Handb. Exp. Pharmacol. 2009, 191, 277– 308 DOI: 10.1007/978-3-540-68964-5_13
  15
  NO-independent, haem-dependent soluble guanylate cyclase stimulators
  Stasch, Johannes-Peter; Hobbs, Adrian J.
  Handbook of Experimental Pharmacology (2009), 191 (cGMP: Generators, Effectors and Therapeutic Implications), 277-308CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
  A review. The nitric oxide (NO) signalling pathway is altered in cardiovascular diseases, including systemic and pulmonary hypertension, stroke, and atherosclerosis. The vasodilatory properties of NO have been exploited for over a century in cardiovascular disease, but NO donor drugs and inhaled NO are assocd. with significant shortcomings, including resistance to NO in some disease states, the development of tolerance during long-term treatment, and non-specific effects such as post-translational modification of proteins. The development of pharmacol. agents capable of directly stimulating the NO receptor, sol. guanylate cyclase (sGC), is therefore highly desirable. The benzylindazole compd. YC-1 was the first sGC stimulator to be identified; this compd. formed a lead structure for the development of optimized sGC stimulators with improved potency and specificity for sGC, including CFM-1571, BAY 41-2272, BAY 41-8543, and BAY 63-2521. In contrast to the NO- and haem-independent sGC activators such as BAY 58-2667, these compds. stimulate sGC activity independent of NO and also act in synergy with NO to produce anti-aggregatory, anti-proliferative, and vasodilatory effects. Recently, aryl-acrylamide compds. were identified independent of YC-1 as sGC stimulators; although structurally dissimilar to YC-1, they have a similar mode of action and promote smooth muscle relaxation. Pharmacol. stimulators of sGC may be beneficial in the treatment of a range of diseases, including systemic and pulmonary hypertension, heart failure, atherosclerosis, erectile dysfunction, and renal fibrosis. An sGC stimulator, BAY 63-2521, is currently in clin. development as an oral therapy for patients with pulmonary hypertension. It has demonstrated efficacy in a proof-of-concept study, reducing pulmonary vascular resistance and increasing cardiac output from baseline. A full, phase 2 trial of BAY 63-2521 in pulmonary hypertension is underway.
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16. 16
  Schmidt, H. H.; Schmidt, P. M.; Stasch, J. P. NO- and haem-independent soluble guanylate cyclase activators Handb. Exp. Pharmacol. 2009, 191, 309– 339 DOI: 10.1007/978-3-540-68964-5_14
  16
  NO- and haem-independent soluble guanylate cyclase activators
  Schmidt, Harald H. H. W.; Schmidt, Peter M.; Stasch, Johannes-Peter
  Handbook of Experimental Pharmacology (2009), 191 (cGMP: Generators, Effectors and Therapeutic Implications), 309-339CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
  A review. Oxidative stress, a risk factor for several cardiovascular disorders, interferes with the NO/sGC/cGMP signalling pathway through scavenging of NO and formation of the strong intermediate oxidant, peroxynitrite. Under these conditions, endothelial and vascular dysfunction develops, culminating in different cardio-renal and pulmonary-vascular diseases. Substituting NO with org. nitrates that release NO (NO donors) has been an important principle in cardiovascular therapy for more than a century. However, the development of nitrate tolerance limits their continuous clin. application and, under oxidative stress and increased formation of peroxynitrite foils the desired therapeutic effect. To overcome these obstacles of nitrate therapy, direct NO- and haem-independent sGC activators have been developed, such as BAY 58-2667 (cinaciguat) and HMR1766 (ataciguat), showing unique biochem. and pharmacol. properties. Both compds. are capable of selectively activating the oxidized/haem-free enzyme via binding to the enzyme's haem pocket, causing pronounced vasodilatation. The potential importance of these new drugs resides in the fact that they selectively target a modified state of sGC that is prevalent under disease conditions as shown in several animal models and human disease. Activators of sGC may be beneficial in the treatment of a range of diseases including systemic and pulmonary hypertension (PH), heart failure, atherosclerosis, peripheral arterial occlusive disease (PAOD), thrombosis and renal fibrosis. The sGC activator HMR1766 is currently in clin. development as an oral therapy for patients with PAOD. The sGC activator BAY 58-2667 has demonstrated efficacy in a proof-of-concept study in patients with acute decompensated heart failure (ADHF), reducing pre- and afterload and increasing cardiac output from baseline. A phase IIb clin. study for the indication of ADHF is currently underway.
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17. 17
  Stasch, J.-P.; Evgenov, O. V. Soluble guanylate cyclase stimulators in pulmonary hypertension Handb. Exp. Pharmacol. 2013, 218, 279– 313 DOI: 10.1007/978-3-662-45805-1_12
  17
  Soluble Guanylate Cyclase Stimulators in Pulmonary Hypertension
  Stasch, Johannes-Peter; Evgenov, Oleg V.
  Handbook of Experimental Pharmacology (2013), 218 (Pharmacotherapy of Pulmonary Hypertension), 279-313CODEN: HEPHD2; ISSN:0171-2004. (Springer GmbH)
  A review. Sol. guanylate cyclase (sGC) is a key enzyme in the nitric oxide (NO) signalling pathway. On binding of NO to its prosthetic heme group, sGC catalyzes the synthesis of the second messenger cyclic guanosine monophosphate (cGMP), which promotes vasodilation and inhibits smooth muscle proliferation, leukocyte recruitment, platelet aggregation and vascular remodelling through a no. of downstream mechanisms. The central role of the NO-sGC-cGMP pathway in regulating pulmonary vascular tone is demonstrated by the dysregulation of NO prodn., sGC activity and cGMP degrdn. in pulmonary hypertension (PH). The sGC stimulators are novel pharmacol. agents that directly stimulate sGC, both independently of NO and in synergy with NO. Optimization of the first sGC stimulator, YC-1, led to the development of the more potent and more specific sGC stimulators, BAY 41-2272, BAY 41-8543 and riociguat (BAY 63-2521). Other sGC stimulators include CFM-1571, BAY 60-4552, vericiguat (BAY 1021189), the acrylamide analog A-350619 and the aminopyrimidine analogs. BAY 41-2272, BAY 41-8543 and riociguat induced marked dose-dependent redns. in mean pulmonary arterial pressure and vascular resistance with a concomitant increase in cardiac output, and they also reversed vascular remodelling and right heart hypertrophy in several exptl. models of PH. Riociguat is the first sGC stimulator that has entered clin. development. Clin. trials have shown that it significantly improves pulmonary vascular haemodynamics and increases exercise ability in patients with pulmonary arterial hypertension (PAH), chronic thromboembolic PH and PH assocd. with interstitial lung disease. Furthermore, riociguat reduces mean pulmonary arterial pressure in patients with PH assocd. with chronic obstructive pulmonary disease and improves cardiac index and pulmonary vascular resistance in patients with PH assocd. with left ventricular systolic dysfunction. These promising results suggest that sGC stimulators may constitute a valuable new therapy for PH. Other trials of riociguat are in progress, including a study of the haemodynamic effects and safety of riociguat in patients with PH assocd. with left ventricular diastolic dysfunction, and long-term extensions of the phase 3 trials investigating the efficacy and safety of riociguat in patients with PAH and chronic thromboembolic PH. Finally, sGC stimulators may also have potential therapeutic applications in other diseases, including heart failure, lung fibrosis, scleroderma and sickle cell disease.
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18. 18
  Ghofrani, H.-A.; Galie, N.; Grimminger, F.; Gruenig, E.; Humbert, M.; Jing, Z.-C.; Keogh, A. M.; Langleben, D.; Kilama, M. O.; Fritsch, A.; Neuser, D.; Rubin, L. J. Riociguat for the treatment of pulmonary arterial hypertension N. Engl. J. Med. 2013, 369, 330– 340 DOI: 10.1056/NEJMoa1209655
  18
  Riociguat for the treatment of pulmonary arterial hypertension
  Ghofrani, Hossein-Ardeschir; Galie, Nazzareno; Grimminger, Friedrich; Gruenig, Ekkehard; Humbert, Marc; Jing, Zhi-Cheng; Keogh, Anne M.; Langleben, David; Kilama, Michael Ochan; Fritsch, Arno; Neuser, Dieter; Rubin, Lewis J.
  New England Journal of Medicine (2013), 369 (4), 330-340CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
  BACKGROUND: Riociguat, a sol. guanylate cyclase stimulator, has been shown in a phase 2 trial to be beneficial in the treatment of pulmonary arterial hypertension. METHODS: In this phase 3, double-blind study, we randomly assigned 443 patients with symptomatic pulmonary arterial hypertension to receive placebo, riociguat in individually adjusted doses of up to 2.5 mg three times daily (2.5 mg-max. group), or riociguat in individually adjusted doses that were capped at 1.5 mg three times daily (1.5 mg-max. group). The 1.5 mg-max. group was included for exploratory purposes, and the data from that group were analyzed descriptively. Patients who were receiving no other treatment for pulmonary arterial hypertension and patients who were receiving endothelin-receptor antagonists or (nonintravenous) prostanoids were eligible. The primary end point was the change from baseline to the end of week 12 in the distance walked in 6 min. Secondary end points included the change in pulmonary vascular resistance, N-terminal pro-brain natriuretic peptide (NT-proBNP) levels, World Health Organization (WHO) functional class, time to clin. worsening, score on the Borg dyspnea scale, quality-of-life variables, and safety. RESULTS: By week 12, the 6-min walk distance had increased by a mean of 30 m in the 2.5 mg-max. group and had decreased by a mean of 6 m in the placebo group (least-squares mean difference, 36 m; 95% confidence interval, 20 to 52; P<0.001). Prespecified subgroup analyses showed that riociguat improved the 6-min walk distance both in patients who were receiving no other treatment for the disease and in those who were receiving endothelin-receptor antagonists or prostanoids. There were significant improvements in pulmonary vascular resistance (P<0.001), NT-proBNP levels (P<0.001), WHO functional class (P = 0.003), time to clin. worsening (P = 0.005), and Borg dyspnea score (P = 0.002). The most common serious adverse event in the placebo group and the 2.5 mg-max. group was syncope (4% and 1%, resp.). CONCLUSIONS: Riociguat significantly improved exercise capacity and secondary efficacy end points in patients with pulmonary arterial hypertension.
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19. 19
  Ghofrani, H.-A.; D’Armini, A. M.; Grimminger, F.; Hoeper, M. M.; Jansa, P.; Kim, N. H.; Mayer, E.; Simonneau, G.; Wilkins, M. R.; Fritsch, A.; Neuser, D.; Weimann, G.; Wang, C. Riociguat for the treatment of chronic thromboembolic pulmonary hypertension N. Engl. J. Med. 2013, 369, 319– 329 DOI: 10.1056/NEJMoa1209657
  19
  Riociguat for the treatment of chronic thromboembolic pulmonary hypertension
  Ghofrani, Hossein-Ardeschir; D'Armini, Andrea M.; Grimminger, Friedrich; Hoeper, Marius M.; Jansa, Pavel; Kim, Nick H.; Mayer, Eckhard; Simonneau, Gerald; Wilkins, Martin R.; Fritsch, Arno; Neuser, Dieter; Weimann, Gerrit; Wang, Chen
  New England Journal of Medicine (2013), 369 (4), 319-329CODEN: NEJMAG; ISSN:0028-4793. (Massachusetts Medical Society)
  BACKGROUND Riociguat, a member of a new class of compds. (sol. guanylate cyclase stimulators), has been shown in previous clin. studies to be beneficial in the treatment of chronic thromboembolic pulmonary hypertension. METHODS In this phase 3, multicenter, randomized, double-blind, placebo-controlled study, we randomly assigned 261 patients with inoperable chronic thromboembolic pulmonary hypertension or persistent or recurrent pulmonary hypertension after pulmonary endarterectomy to receive placebo or riociguat. The primary end point was the change from baseline to the end of week 16 in the distance walked in 6 min. Secondary end points included changes from baseline in pulmonary vascular resistance, N-terminal pro-brain natriuretic peptide (NT-proBNP) level, World Health Organization (WHO) functional class, time to clin. worsening, Borg dyspnea score, quality-of-life variables, and safety. RESULTS By week 16, the 6-min walk distance had increased by a mean of 39 m in the riociguat group, as compared with a mean decrease of 6 m in the placebo group (least-squares mean difference, 46 m; 95% confidence interval [CI], 25 to 67; P<0.001). Pulmonary vascular resistance decreased by 226 dyn·sec·cm-5 in the riociguat group and increased by 23 dyn·sec·cm-5 in the placebo group (least-squares mean difference, -246 dyn·sec·cm-5; 95% CI, -303 to -190; P<0.001). Riociguat was also assocd. with significant improvements in the NT-proBNP level (P<0.001) and WHO functional class (P = 0.003). The most common serious adverse events were right ventricular failure (in 3% of patients in each group) and syncope (in 2% of the riociguat group and in 3% of the placebo group). CONCLUSIONS Riociguat significantly improved exercise capacity and pulmonary vascular resistance in patients with chronic thromboembolic pulmonary hypertension.
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20. 20
  Hambly, N.; Granton, J. Riociguat for the treatment of pulmonary hypertension Expert Rev. Respir. Med. 2015, 9, 679– 695 DOI: 10.1586/17476348.2015.1106316
  20
  Riociguat for the treatment of pulmonary hypertension
  Hambly, Nathan; Granton, John
  Expert Review of Respiratory Medicine (2015), 9 (6), 679-695CODEN: ERRMBF; ISSN:1747-6348. (Taylor & Francis Ltd.)
  A review. Nitric oxide (NO) is a crit. signaling mol. in the pulmonary vasculature. NO activates sol. guanylate cyclase (sGC) resulting in the synthesis of cGMP - a key mediator of pulmonary artery vasodilatation that may also inhibit smooth muscle proliferation and platelet aggregation. Pulmonary hypertension, a serious, progressive and often fatal disease is characterized by NO-sGC-sGMP pathway dysregulation. Riociguat is a member of a novel therapeutic class known as sol. guanylate stimulators. Riociguat has a dual mode of action, acting in synergy with endogenous NO and also directly stimulating sGC independently of NO availability. Phase 3 randomized control trials have demonstrated that riociguat improves clin., physiol. and hemodynamic parameters in patients with pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. In this review we will discuss the pharmacol. properties of riociguat and its appropriate implementation into clin. practice.
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21. 21
  Frey, R.; Mueck, W.; Unger, S.; Artmeier-Brandt, U.; Weimann, G.; Wensing, G. Single-dose pharmacokinetics, pharmacodynamics, tolerability, and safety of the soluble guanylate cyclase stimulator BAY 63–2521: an ascending-dose study in healthy male volunteers J. Clin. Pharmacol. 2008, 48, 926– 934 DOI: 10.1177/0091270008319793
  21
  Single-dose pharmacokinetics, pharmacodynamics, tolerability, and safety of the soluble guanylate cyclase stimulator BAY 63-2521: an ascending-dose study in healthy male volunteers
  Frey, Reiner; Mueck, Wolfgang; Unger, Sigrun; Artmeier-Brandt, Ulrike; Weimann, Gerrit; Wensing, Georg
  Journal of Clinical Pharmacology (2008), 48 (8), 926-934CODEN: JCPCBR; ISSN:0091-2700. (Sage Publications)
  The aim of the study was to assess the safety, tolerability, pharmacokinetics, and pharmacodynamics of BAY 63-2521, a new drug in development for pulmonary hypertension. Fifty-eight healthy male volunteers received a single oral dose of BAY 63-2521 (0.25-5 mg) or placebo. No serious adverse events were reported; there were no life-threatening events. Heart rate over 1 min, an indicator of the effect of a vasodilating agent on the cardiovascular system in healthy subjects, was increased dose dependently vs. placebo at BAY 63-2521 doses of 1 to 5 mg (P < .01). Mean arterial and diastolic pressures were decreased vs. placebo at doses of 1 mg (P < .05) and 5 mg (P < .01). Systolic pressure was not significantly affected. BAY 63-2521 was readily absorbed and exhibited dose-proportional pharmacokinetics. The pharmacodynamic and pharmacokinetic properties of BAY 63-2521 suggest that it can offer a unique mode of action in the treatment of pulmonary hypertension.
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22. 22
  Gheorghiade, M.; Greene, S. J.; Butler, J.; Filippatos, G.; Lam, C. S.; Maggioni, A. P.; Ponikowski, P.; Shah, S. J.; Solomon, S. D.; Kraigher-Krainer, E.; Samano, E. T.; Müller, K.; Roessig, L.; Pieske, B. Effect of vericiguat, a soluble guanylate cyclase stimulator, on natriuretic peptide levels in patients with worsening chronic heart failure and reduced ejection fraction: the SOCRATES-REDUCED randomized trial JAMA, J. Am. Med. Assoc. 2015, 314, 2251– 2262 DOI: 10.1001/jama.2015.15734
  There is no corresponding record for this reference.
23. 23
  Pieske, B.; Butler, J.; Filippatos, G.; Lam, C.; Maggioni, A. P.; Ponikowski, P.; Shah, S.; Solomon, S.; Kraigher-Krainer, E.; Samano, E. T.; Scalise, A. V.; Mueller, K.; Roessig, L.; Gheorghiade, M. Rationale and design of the SOluble guanylate Cyclase stimulatoR in heArT failurE Studies (SOCRATES) Eur. J. Heart Failure 2014, 16, 1026– 38 DOI: 10.1002/ejhf.135
  23
  Rationale and design of the SOluble guanylate Cyclase stimulatoR in heArT failurE Studies (SOCRATES)
  Pieske, Burkert; Butler, Javed; Filippatos, Gerasimos; Lam, Carolyn; Maggioni, Aldo Pietro; Ponikowski, Piotr; Shah, Sanjiv; Solomon, Scott; Kraigher-Krainer, Elisabeth; Samano, Eliana Tibana; Scalise, Andrea Viviana; Mueller, Katharina; Roessig, Lothar; Gheorghiade, Mihai; N/A
  European Journal of Heart Failure (2014), 16 (9), 1026-1038CODEN: EJHFFS; ISSN:1388-9842. (John Wiley & Sons Ltd.)
  Aims : The clin. outcomes for patients with worsening chronic heart failure (WCHF) remain exceedingly poor despite contemporary evidence-based therapies, and effective therapies are urgently needed. Accumulating evidence supports augmentation of cyclic guanosine monophosphate (cGMP) signalling as a potential therapeutic strategy for HF with reduced or preserved ejection fraction (HFrEF and HFpEF, resp.). Direct sol. guanylate cyclase (sGC) stimulators target reduced cGMP generation due to insufficient sGC stimulation and represent a promising method for cGMP enhancement. Methods : The phase II Sol. guanylate Cyclase stimulatoR in heArT failurE Study (SOCRATES) program consists of two randomized, parallel-group, placebo-controlled, double-blind, multicentre studies, SOCRATES-REDUCED (in patients with LVEF <45%) and SOCRATES-PRESERVED (in those with LVEF ≥45%), that will explore the pharmacodynamic effects, safety and tolerability, and pharmacokinetics of four dose regimens of the once-daily oral sGC stimulator vericiguat (BAY 1021189) over 12 wk compared with placebo. These studies will enrol patients stabilized during hospitalization for HF at the time of discharge or within 4 wk thereafter. The primary endpoint in SOCRATES-REDUCED is change in NT-proBNP at 12 wk. The primary endpoints in SOCRATES-PRESERVED are change in NT-proBNP and left atrial vol. at 12 wk. Perspectives : SOCRATES will be the first program to enrol specifically both inpatients and outpatients with WCHF and patients with reduced or preserved ejection fraction. Results will inform the benefits of pursuing subsequent event-driven clin. outcome trials with sGC stimulators in this patient population. Trial registration : NCT01951625 (SOCRATES-REDUCED) and NCT01951638 (SOCRATES-PRESERVED).
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24. 24
  Clinicaltrials.gov. Identifier: NCT02861534 (accessed August 5, 2016).
  There is no corresponding record for this reference.
25. 25
  Mittendorf, J.; Weigand, S.; Alonso-Alija, C.; Bischoff, E.; Feurer, A.; Gerisch, M.; Kern, A.; Knorr, A.; Lang, D.; Muenter, K.; Radtke, M.; Schirok, H.; Schlemmer, K.-H.; Stahl, E.; Straub, A.; Wunder, F.; Stasch, J.-P. Discovery of riociguat (BAY 63–2521): a potent, oral stimulator of soluble guanylate cyclase for the treatment of pulmonary hypertension ChemMedChem 2009, 4, 853– 865 DOI: 10.1002/cmdc.200900014
  25
  Discovery of Riociguat (BAY 63-2521): A Potent, Oral Stimulator of Soluble Guanylate Cyclase for the Treatment of Pulmonary Hypertension
  Mittendorf, Joachim; Weigand, Stefan; Alonso-Alija, Cristina; Bischoff, Erwin; Feurer, Achim; Gerisch, Michael; Kern, Armin; Knorr, Andreas; Lang, Dieter; Muenter, Klaus; Radtke, Martin; Schirok, Hartmut; Schlemmer, Karl-Heinz; Stahl, Elke; Straub, Alexander; Wunder, Frank; Stasch, Johannes-Peter
  ChemMedChem (2009), 4 (5), 853-865CODEN: CHEMGX; ISSN:1860-7179. (Wiley-VCH Verlag GmbH & Co. KGaA)
  Direct stimulation of sol. guanylate cyclase (sGC),a key signal-transduction enzyme activated by nitric oxide (NO), represents a promising therapeutic strategy for the treatment of a range of diseases, including the severely disabling pulmonary hypertension (PH). Impairments of the NO-sGC signaling pathway have been implicated in the pathogenesis of cardiovascular and other diseases. Optimization of the unfavorable drug metab. and pharmacokinetic (DMPK) profile of previous sGC stimulators provided riociguat I, which is currently being investigated in phase III clin. trials for the oral treatment of PH. The compd. I exhibited an improved DMPK profile and exerted strong effects on pulmonary hemodynamics and exercise capacity in patients with PH.
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26. 26
  Gnoth, M. J.; Hopfe, P. M.; Czembor, W. Determination of riociguat and its major human metabolite M-1 in human plasma by stable-isotope dilution LCMS/MS Bioanalysis 2015, 7, 193– 205 DOI: 10.4155/bio.14.257
  There is no corresponding record for this reference.
27. 27
  Becker, C.; Frey, R.; Thomas, D.; Reber, M.; Weimann, G.; Arens, E. R.; Mück, W.; Unger, S.; Dietrich, H. Pharmacokinetic interaction of riociguat with ketoconazole, clarithromycin, and midazolam Pulm. Circ. 2016, 6, S49– 57 DOI: 10.1086/685016
  27
  Pharmacokinetic interaction of riociguat with ketoconazole, clarithromycin, and midazolam
  Becker, Corina; Frey, Reiner; Unger, Sigrun; Thomas, Dirk; Reber, Michael; Weimann, Gerrit; Dietrich, Hartmut; Arens, Erich R.; Mueck, Wolfgang
  Pulmonary Circulation (2016), 6 (Suppl.1), S49-S57CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
  Riociguat is a sol. guanylate cyclase stimulator for the treatment of pulmonary hypertension that is principally metabolized via the cytochrome P 450 (CYP) pathway. Three studies in healthy males investigated potential pharmacokinetic interactions between riociguat and CYP inhibitors (ketoconazole, clarithromycin, and midazolam). In two studies, subjects were pretreated with either oncedaily ketoconazole 400 mg or twice-daily clarithromycin 500 mg for 4 days before cotreatment with either riociguat 0.5 mg ± ketoconazole 400 mg or riociguat 1.0 mg ± clarithromycin 500 mg. In the third study, subjects received riociguat 2.5 mg 3 times daily (tid) for 3 days, followed by cotreatment with riociguat 2.5 mg tid ± midazolam 7.5 mg. Pharmacokinetic parameters, the effect of smoking on riociguat pharmacokinetics, safety, and tolerability were assessed. Pre- and cotreatment with ketoconazole and clarithromycin led to increased riociguat exposure. Pre- and cotreatment with riociguat had no significant effect on midazolam plasma concns. In all studies, the bioavailability of riociguat was reduced in smokers because its clearance to the metabolite M1 increased. Riociguat ± ketoconazole, clarithromycin, or midazolam was generally well tolerated. The most common treatment-emergent adverse events (TEAEs) across all studies were headache and dyspepsia. One serious TEAE was reported in the midazolam study. Owing to the potential for hypotension, concomitant use of riociguat with multipathway inhibitors, such as ketoconazole, should be approached with caution. Coadministration of riociguat with strong CYP3A4 inhibitors, for example, clarithromycin, does not require addnl. dose adjustment. No significant drugdrug interaction was revealed between riociguat and midazolam.
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28. 28
  Frey, R.; Becker, C.; Unger, S.; Schmidt, A.; Wensing, G.; Mück, W. Assessment of the effects of renal impairment and smoking on the pharmacokinetics of a single oral dose of the soluble guanylate cyclase stimulator riociguat (BAY 63–2521) Pulm. Circ. 2016, 6, S15– 26 DOI: 10.1086/685017
  28
  Assessment of the effects of renal impairment and smoking on the pharmacokinetics of a single oral dose of the soluble guanylate cyclase stimulator riociguat (BAY 63-2521)
  Frey, Reiner; Becker, Corina; Unger, Sigrun; Schmidt, Anja; Wensing, Georg; Mueck, Wolfgang
  Pulmonary Circulation (2016), 6 (Suppl.1), S15-S26CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
  Renal impairment is a common comborbidity in patients with pulmonary hypertension. The breakdown of riociguat, an oral sol. guanylate cyclase stimulator used to treat pulmonary hypertension, may be affected by smoking because polycyclic arom. hydrocarbons in tobacco smoke induce expression of one of the metabolizing enzymes, CYP1A1. Two nonrandomized, nonblinded studies were therefore performed to investigate the pharmacokinetics and safety of a single oral dose of riociguat 1.0 mg in individuals with mild, moderate, or severe renal impairment compared with age-, wt.-, and sex-matched healthy controls, including either smokers and nonsmokers (study I) or nonsmokers alone (study II). Pharmacokinetic analyses focused on the integrated per-protocol data set of both studies (N = 63). In patients with renal impairment, the renal clearance of riociguat was reduced and its terminal half-life prolonged compared with those in healthy controls. There was a monotonic relationship between creatinine clearance on treatment day and riociguat renal clearance (R2 = 0.62). However, increased riociguat exposure with decreasing renal function was not strictly proportional. Riociguat exposure appeared to be greater in nonsmokers than in the combined population of smokers and nonsmokers, irresp. of renal function. Adverse events were mild to moderate and in line with the mode of action of riociguat. No serious adverse events occurred. In conclusion, renal impairment was assocd. with reduced riociguat clearance compared with that in controls; however, riociguat exposure in patients with renal impairment was highly variable, and ranges overlapped with those obsd. in healthy controls.
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29. 29
  Saleh, S.; Frey, R.; Becker, C.; Unger, S.; Wensing, G.; Mück, W. Bioavailability, pharmacokinetics, and safety of riociguat given as an oral suspension or crushed tablet with and without food Pulm. Circ. 2016, 6, S66– 74 DOI: 10.1086/685020
  29
  Bioavailability, pharmacokinetics, and safety of riociguat given as an oral suspension or crushed tablet with and without food
  Saleh, Soundos; Frey, Reiner; Becker, Corina; Unger, Sigrun; Wensing, Georg; Mueck, Wolfgang
  Pulmonary Circulation (2016), 6 (Suppl.1), S66-S74CODEN: PCUIAX; ISSN:2045-8940. (University of Chicago Press)
  Riociguat is approved for the treatment of pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension. Some patients have difficulty swallowing tablets; therefore, 2 randomized, nonblinded, crossover studies compared the relative bioavailability of riociguat oral suspensions and immediate-release (IR) tablet and of crushed-tablet prepns. vs. whole IR tablet. In study 1, 30 healthy subjects received 5 single riociguat doses: 0.3 and 2.4 mg (0.15 mg/mL suspensions), 0.15 mg (0.03 mg/mL), and 1.0 mg (whole IR tablet) under fasted conditions and 2.4 mg (0.15 mg/mL) after a high-fat, high-calorie American-style breakfast. In study 2, 25 healthy men received 4 single 2.5-mg doses: whole IR tablet and crushed IR tablet suspended in applesauce and water, resp., under fasted conditions, and whole IR tablet after a continental breakfast. In study 1, dose-normalized pharmacokinetics of riociguat oral suspensions and 1.0-mg whole IR tablet were similar in fasted conditions; 90% confidence intervals for riociguat area under the curve (AUC) to dose and mean max. concn. (Cmax) to dose were within bioequivalence criteria. After food, dosenormalized AUC and Cmax decreased by 15% and 38%, resp. In study 2, riociguat exposure was similar for all prepns.; AUC ratios for crushed-IR-tablet prepns. to whole IR tablet were within bioequivalence criteria. The Cmax increased by 17% for crushed IR tablet in water vs. whole IR tablet. Food intake decreased Cmax of the whole tablet by 16%, with unaltered AUC vs. fasted conditions. Riociguat bioavailability was similar between the oral suspensions and the whole IR tablet; exposure was similar between whole IR tablet and crushed-IR-tablet prepns. Minor food effects were obsd. Results suggest that riociguat formulations are interchangeable.
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30. 30
  Zhao, X.; Wang, Z.; Wang, Y.; Zhang, H.; Blode, H.; Yoshikawa, K.; Becker, C.; Unger, S.; Frey, R.; Cui, Y. Pharmacokinetics of the soluble guanylate cyclase stimulator riociguat in healthy young Chinese male non-smokers and smokers: results of a randomized, double-blind, placebo-controlled study Clin. Pharmacokinet. 2016, 55, 615– 624 DOI: 10.1007/s40262-015-0337-4
  30
  Pharmacokinetics of the Soluble Guanylate Cyclase Stimulator Riociguat in Healthy Young Chinese Male Non-Smokers and Smokers: Results of a Randomized, Double-Blind, Placebo-Controlled Study
  Zhao, Xia; Wang, Zining; Wang, Yukun; Zhang, Hong; Blode, Hartmut; Yoshikawa, Kenichi; Becker, Corina; Unger, Sigrun; Frey, Reiner; Cui, Yimin
  Clinical Pharmacokinetics (2016), 55 (5), 615-624CODEN: CPKNDH; ISSN:0312-5963. (Springer International Publishing AG)
  Background and Objectives: The aim of this study was to investigate the pharmacokinetics, safety, and tolerability of riociguat after single and multiple oral doses of 1 or 2 mg three times daily (tid), and to det. the effect of smoking on riociguat pharmacokinetics in Chinese men. Methods: In a randomized, double-blind, placebo-controlled, single-center study stratified for smokers and non-smokers, healthy Chinese men aged 18-45 years received two riociguat doses: Dose Step 1 (1 mg) then Dose Step 2 (2 mg) conducted after the safety and tolerability at Dose Step 1 was confirmed. For each step, 12 subjects received riociguat and six received placebo. A single dose was given on Day 1, followed by a 48-h pharmacokinetic profile. Multiple-dose treatment tid was then given for 6 days (Days 3-8), with a last single dose on Day 9, followed by a 72-h pharmacokinetic profile. Primary outcomes were pharmacokinetic parameters for riociguat after single and multiple dosing. Results: Thirty-six subjects (18 smokers; 18 non-smokers) were randomized and provided valid pharmacokinetic data. Riociguat and its pharmacol. active metabolite M1 (BAY 60-4552) showed nearly dose-proportional pharmacokinetics. Accumulation was minimal in smokers and approx. two-fold in non-smokers. Exposure for riociguat was decreased by ≥60 % in smokers. No serious or significant adverse events occurred during the study. Conclusions: Riociguat pharmacokinetics showed dose proportionality in healthy Chinese men, as previously demonstrated in healthy white male individuals. Exposure to riociguat was substantially decreased in smokers compared with non-smokers. Riociguat was well tolerated in Chinese men.
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31. 31
  Highlights of Prescribing Information: Riociguat (Adempas); U.S. Food and Drug Administration: Silver Spring, MD, 2014; http://www.accessdata.fda.gov/drugsatfda_docs/label/2014/204819s002lbl.pdf (accessed April 4, 2014).
  There is no corresponding record for this reference.
32. 32
  Bitterli, P.; Erlenmeyer, H. Some derivatives of triazolopyrimidine Helv. Chim. Acta 1951, 34, 835– 840 DOI: 10.1002/hlca.19510340311
  There is no corresponding record for this reference.
33. 33
  Reinecke, M. G.; Woodrow, T. A.; Brown, E. S. Pyrazolo[3,4-c]pyridazines from hydrazine and aminothiophenecarboxylates J. Org. Chem. 1992, 57, 1018– 1021 DOI: 10.1021/jo00029a046
  There is no corresponding record for this reference.
34. 34
  Wunder, F.; Stasch, J.-P.; Hütter, J.; Alonso-Alija, C.; Hüser, J.; Lohrmann, E. A cell-based cGMP assay useful for ultra-high-throughput screening and identification of modulators of the nitric oxide/cGMP pathway Anal. Biochem. 2005, 339, 104– 112 DOI: 10.1016/j.ab.2004.12.025
  34
  A cell-based cGMP assay useful for ultra-high-throughput screening and identification of modulators of the nitric oxide/cGMP pathway
  Wunder, Frank; Stasch, Johannes-Peter; Huetter, Joachim; Alonso-Alija, Cristina; Hueser, Joerg; Lohrmann, Emanuel
  Analytical Biochemistry (2005), 339 (1), 104-112CODEN: ANBCA2; ISSN:0003-2697. (Elsevier)
  We have established a rapid, homogeneous, cell-based, and highly sensitive assay for guanosine 3'-5'-cyclic monophosphate (cGMP) that is suitable for fully automated ultra-high-throughput screening. In this assay system, cGMP prodn. is monitored in living cells via Ca2+ influx through the olfactory cyclic nucleotide-gated cation channel CNGA2, acting as the intracellular cGMP sensor. A stably transfected Chinese hamster ovary (CHO) cell line was generated recombinantly expressing sol. guanylate cyclase, CNGA2, and aequorin as a luminescence indicator for the intracellular calcium concn. This cell line was used to screen more than 900,000 compds. in an automated ultra-high-throughput screening assay using 1536-well microtiter plates. In this way, we have been able to identify BAY 58-2667, a member of a new class of amino dicarboxylic acids that directly activate sol. guanylate cyclase. The assay system allows the real-time cGMP detection within living cells and makes it possible to screen for activators and inhibitors of enzymes involved in the nitric oxide/cGMP pathway.
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35. 35
  Hillisch, A.; Heinrich, N.; Wild, H. Computational chemistry in the pharmaceutical industry: from childhood to adolescence ChemMedChem 2015, 10, 1958– 1962 DOI: 10.1002/cmdc.201500346
  35
  Computational Chemistry in the Pharmaceutical Industry: From Childhood to Adolescence
  Hillisch, Alexander; Heinrich, Nikolaus; Wild, Hanno
  ChemMedChem (2015), 10 (12), 1958-1962CODEN: CHEMGX; ISSN:1860-7179. (Wiley-VCH Verlag GmbH & Co. KGaA)
  Computational chem. within the pharmaceutical industry has grown into a field that proactively contributes to many aspects of drug design, including target selection and lead identification and optimization. While methodol. advancements have been key to this development, organizational developments have been crucial to our success as well. In particular, the interaction between computational and medicinal chem. and the integration of computational chem. into the entire drug discovery process have been invaluable. Over the past ten years we have shaped and developed a highly efficient computational chem. group for small-mol. drug discovery at Bayer HealthCare that has significantly impacted the clin. development pipeline. In this article we describe the setup and tasks of the computational group and discuss external collaborations. We explain what we have found to be the most valuable and productive methods and discuss future directions for computational chem. method development. We share this information with the hope of igniting interesting discussions around this topic.
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36. 36
  Burkhard, J. A.; Wuitschik, G.; Rogers-Evans, M.; Müller, K.; Carreira, E. M. Oxetanes as versatile elements in drug discovery and synthesis Angew. Chem., Int. Ed. 2010, 49, 9052– 9067 DOI: 10.1002/anie.200907155
  36
  Oxetanes as Versatile Elements in Drug Discovery and Synthesis
  Burkhard, Johannes A.; Wuitschik, Georg; Rogers-Evans, Mark; Mueller, Klaus; Carreira, Erick M.
  Angewandte Chemie, International Edition (2010), 49 (48), 9052-9067CODEN: ACIEF5; ISSN:1433-7851. (Wiley-VCH Verlag GmbH & Co. KGaA)
  A review. Sizable resources, both financial and human, are invested each year in the development of new pharmaceutical agents. However, despite improved techniques, the new compds. often encounter difficulties in satisfying and overcoming the numerous physicochem. and many pharmacol. constraints and hurdles. Oxetanes have been shown to improve key properties when grafted onto mol. scaffolds. Of particular interest are oxetanes that are substituted only in the 3-position, since such units remain achiral and their introduction into a mol. scaffold does not create a new stereocenter. This Minireview gives an overview of the recent advances made in the prepn. and use of 3-substituted oxetanes. It also includes a discussion of the site-dependent modifications of various physicochem. and biochem. properties that result from the incorporation of the oxetane unit in mol. architectures.
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37. 37
  Wuitschik, G.; Carreira, E. M.; Wagner, B.; Fischer, H.; Parrilla, I.; Schuler, F.; Rogers-Evans, M.; Müller, K. Oxetanes in drug discovery: structural and synthetic insights J. Med. Chem. 2010, 53, 3227– 3246 DOI: 10.1021/jm9018788
  37
  Oxetanes in Drug Discovery: Structural and Synthetic Insights
  Wuitschik, Georg; Carreira, Erick M.; Wagner, Bjorn; Fischer, Holger; Parrilla, Isabelle; Schuler, Franz; Rogers-Evans, Mark; Muller, Klaus
  Journal of Medicinal Chemistry (2010), 53 (8), 3227-3246CODEN: JMCMAR; ISSN:0022-2623. (American Chemical Society)
  The use of oxetanes as replacements for gem-di-Me or carbonyl groups and their effects on the aq. soly., lipophilicity, metabolic stability, and conformation for various compds. are studied; methods for the prepn. of a variety of substituted oxetanes are given. The magnitude of changes in properties and in metabolic stability with oxetane substitution depends on the structural context; for example, substitution of a gem-di-Me group with an oxetane, aq. soly. may increase by a factor of 4 to more than 4000 while reducing the rate of metabolic degrdn. in most cases. Incorporation of an oxetane into an aliph. chain increases in some cases the preference for synclinal conformations rather than antiplanar conformations of the chain. Spirocyclic oxetanes such as an oxazaspiroheptane resemble commonly used fragments in drug discovery, such as morpholines, and in some cases increase aq. soly. more effectively than morpholines. An improved chemoselective oxidn. of 3-oxetanol to 3-oxetanone is disclosed; olefination of 3-oxetanone by a variety of methods yields alkylideneoxetanes I [R = (EtO)2P(:O), OHC, O2N, EtO2C, NC, PhO2S, MeCO, 1-(4-chlorophenyl)-1-cyclobutanecarbonyl]. I (R = EtO2C, OHC, O2N) undergo addn. reactions with nucleophiles such as amines, carbonyl compds., and arylboronic acids to give oxetanes such as II. The crystal structures of a variety of oxetanes are detd.
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38. 38
  Leeson, P. D.; Young, R. J. Molecular property design: does everyone get it? ACS Med. Chem. Lett. 2015, 6, 722– 725 DOI: 10.1021/acsmedchemlett.5b00157
  38
  Molecular Property Design: Does Everyone Get It?
  Leeson, Paul D.; Young, Robert J.
  ACS Medicinal Chemistry Letters (2015), 6 (7), 722-725CODEN: AMCLCT; ISSN:1948-5875. (American Chemical Society)
  A review. The principles of mol. property optimization in drug design have been understood for decades, yet much drug discovery activity today is conducted at the periphery of historical druglike property space. Lead optimization trajectories aimed at reducing physicochem. risk, assisted by ligand efficiency metrics, could help to reduce clin. attrition rates.
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39. 39
  Sharkovska, Y.; Kalk, P.; Lawrenz, B.; Godes, M.; Hoffmann, L. S.; Wellkisch, K.; Geschka, S.; Relle, K.; Hocher, B.; Stasch, J.-P. Nitric oxide-independent stimulation of soluble guanylate cyclase reduces organ damage in experimental low-renin and high-renin models J. Hypertens. 2010, 28, 1666– 1675 DOI: 10.1097/HJH.0b013e32833b558c
  39
  Nitric oxide-independent stimulation of soluble guanylate cyclase reduces organ damage in experimental low-renin and high-renin models
  Sharkovska, Yuliya; Kalk, Philipp; Lawrenz, Bettina; Godes, Michael; Hoffmann, Linda Sarah; Wellkisch, Kathrin; Geschka, Sandra; Relle, Katharina; Hocher, Berthold; Stasch, Johannes-Peter
  Journal of Hypertension (2010), 28 (8), 1666-1675CODEN: JOHYD3; ISSN:0263-6352. (Lippincott Williams & Wilkins)
  The nitric oxide-sol. guanylate cyclase (sGC)-cGMP signal transduction pathway is impaired in different cardiovascular diseases, including pulmonary hypertension, heart failure and arterial hypertension. Riociguat is a novel stimulator of sol. guanylate cyclase (sGC). However, little is known about the effects of sGC stimulators in exptl. models of hypertension. We thus investigated the cardio-renal protective effects of riociguat in low-renin and high-renin rat models of hypertension. The vasorelaxant effect of riociguat was tested in vitro on isolated saphenous artery rings of normal and nitrate tolerant rabbits. The cardiovascular in-vivo effects of sGC stimulation were evaluated in hypertensive renin-transgenic rats treated with the nitric oxide-synthase inhibitor N-nitro-L-arginine Me ester (L-NAME) (high-renin model) and in rats with 5/6 nephrectomy (low-renin model). In both animal models, riociguat treatment improved survival and normalized blood pressure. Moreover, in the L-NAME study part, riociguat reduced cardiac target organ damage as indicated by lower plasma ANP, lower relative left ventricular wt. and lower cardiac interstitial fibrosis, and reduced renal target organ damage as indicated by lower plasma creatinine and urea, less glomerulosclerosis and less renal interstitial fibrosis. In the 5/6 nephrectomy study part, riociguat reduced cardiac target organ damage as indicated by lower plasma ANP, lower relative left ventricular wt., lower myocyte diam. and lower arterial media/lm ratio, and reduced renal target organ damage as indicated by improved creatinine clearance and less renal interstitial fibrosis. We demonstrate for the first time that the novel sGC stimulator riociguat shows in 2 independent models of hypertension a potent protection against cardiac and renal target organ damage.
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40. 40
  Dubin, R. F.; Shah, S. J. Soluble guanylate cyclase stimulators: a novel treatment option for heart failure associated with cardiorenal syndromes? Curr. Heart Failure Rep. 2016, 13, 132– 139 DOI: 10.1007/s11897-016-0290-z
  40
  Soluble Guanylate Cyclase Stimulators: a Novel Treatment Option for Heart Failure Associated with Cardiorenal Syndromes?
  Dubin, Ruth F.; Shah, Sanjiv J.
  Current Heart Failure Reports (2016), 13 (3), 132-139CODEN: CHFRCF; ISSN:1546-9549. (Springer)
  Heart failure in the setting of chronic kidney disease (CKD) is an increasingly common scenario and carries a poor prognosis. Clinicians lack tools for primary or secondary heart failure prevention in patients with cardiorenal syndromes. In patients without CKD, angiotensin-converting enzyme inhibitors (ACE-I) or angiotensin receptor blockers (ARB) and statins mitigate cardiovascular risk in large part due to salutary effects on the endothelium. In the setting of CKD, use of these therapies is limited by adverse effects of hyperkalemia in pre-dialysis CKD (ACE-I/ARB), or potential increased risk of stroke in end-stage renal disease (statins). The sol. guanylate cyclase (sGC) stimulators are a novel class of medications that promote endothelial and myocardial function with no known risk of hyperkalemia or stroke. In this review, we discuss the evidence emerging from recent clin. trials of sGC stimulators in pulmonary hypertension and heart failure, the diseased pathways involved in cardiorenal syndromes likely to be restored by sGC stimulators, and several strategies for designing future clin. trials of cardiorenal syndromes that might shorten the timeline for discovery and approval of effective cardiovascular therapies in these high-risk patients.
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41. 41
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Griebenow, N.; Lang, D.; Wunder, F.; Huebsch, W.; Lindner, N.; Vakalopoulos, A.; Tersteegen, A. Preparation of annelated pyrimidine derivatives useful in the treatment and prophylaxis of cardiovascular diseases. WO2013030288, 2013.
  There is no corresponding record for this reference.
42. 42
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Baerfacker, L.; Weigand, S. Bicyclic aza-heterocycles as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012028647, 2012.
  There is no corresponding record for this reference.
43. 43
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Schirok, H.; Jautelat, R. Substituted methyl pyrimidin-5-yl carbamates as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012010578, 2012.
  There is no corresponding record for this reference.
44. 44
  Alonso-Alija, C.; Bischoff, E.; Muenter, K.; Stasch, J.-P.; Stahl, E.; Weigand, S.; Feurer, A. Preparation of [(pyrazolopyridinyl)pyrimidinyl]carbamates stimulating soluble guanylate cyclase for treating cardiovascular diseases and/or sexual dysfunction. WO 2003095451, 2003.
  There is no corresponding record for this reference.
45. 45
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Wunder, F.; Li, V. M.-J.; Mittendorf, J.; Jautelat, R. Carbamate-substituted diaminopyrimidines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012010576, 2012.
  There is no corresponding record for this reference.
46. 46
  Schirok, H.; Mittendorf, J.; Stasch, J.-P.; Wunder, F.; Stoll, F.; Schlemmer, K.-H. Azabicyclic derivatives as stimulators of guanylate cyclase, their preparation, pharmaceutical compositions, and use for the treatment of cardiovascular disorders. WO 2008031513, 2008.
  There is no corresponding record for this reference.
47. 47
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J.; Mittendorf, J.; Schlemmer, K.-H.; Jautelat, R. Substituted 6-fluoro-1H-pyrazolo[4,3-b]pyridines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012059549, 2012.
  There is no corresponding record for this reference.
48. 48
  Follmann, M.; Stasch, J.-P.; Redlich, G.; Straub, A.; Ackerstaff, J.; Griebenow, N.; Knorr, A.; Wunder, F.; Li, V. M.-J. Substituted imidazopyridazines as guanylate cyclase inhibitors and their preparation and use in the treatment of cardiovascular diseases. WO 2012152630, 2012.
  There is no corresponding record for this reference.
49. 49
  Straub, A.; Feurer, A.; Alonso-Alija, C.; Stasch, J.-P.; Perzborn, E.; Huetter, J.; Dembowsky, K.; Stahl, E. Substituted pyrazole derivatives condensed with six-membered heterocyclic rings as cardiovascular agents and their preparation. WO 2000006569, 2000.
  There is no corresponding record for this reference.
50. 50
  Walsky, R. L.; Obach, R. S. Validated assays for human cytochrome P450 activities Drug Metab. Dispos. 2004, 32, 647– 660 DOI: 10.1124/dmd.32.6.647
  50
  Validated assays for human cytochrome P450 activities
  Walsky, Robert L.; Obach, R. Scott
  Drug Metabolism and Disposition (2004), 32 (6), 647-660CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)
  The measurement of the effect of new chem. entities on human cytochrome P 450 marker activities using in vitro experimentation represents an important exptl. approach in drug development. In vitro drug interaction data can be used in guiding the design of clin. drug interaction studies, or, when no effect is obsd. in vitro, the data can be used in place of an in vivo study to claim that no interaction will occur in vivo. To make such a claim, it must be assured that the in vitro expts. are performed with abs. confidence in the methods used and data obtained. To meet this need, 12 semiautomated assays for human P 450 marker substrate activities have been developed and validated using approaches described in the GLP (good lab. practices) as per the code of U.S. Federal Regulations. The assays that were validated are: phenacetin O-deethylase (CYP1A2), coumarin 7-hydroxylase (CYP2A6), bupropion hydroxylase (CYP2B6), amodiaquine N-deethylase (CYP2C8), diclofenac 4'-hydroxylase and tolbutamide methylhydroxylase (CYP2C9), (S)-mephenytoin 4'-hydroxylase (CYP2C19), dextromethorphan O-demethylase (CYP2D6), chlorzoxazone 6-hydroxylase (CYP2E1), felodipine dehydrogenase, testosterone 6β-hydroxylase, and midazolam 1'-hydroxylase (CYP3A4 and CYP3A5). High-pressure liq. chromatog.-tandem mass spectrometry, using stable isotope-labeled internal stds., has been applied as the anal. method. This anal. approach, through its high sensitivity and selectivity, has permitted the use of very low incubation concns. of microsomal protein (0.01-0.2 mg/mL). Anal. assay accuracy and precision values were excellent. Enzyme kinetic and inhibition parameters obtained using these methods demonstrated high precision and were within the range of values previously reported in the scientific literature. These methods should prove useful in the routine assessments of the potential for new drug candidates to elicit pharmacokinetic drug interactions via inhibition of cytochrome P 450 activities.
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51. 51
  Obach, R. S.; Walsky, R. L.; Venkatakrishnan, K. Mechanism-based inactivation of human cytochrome p450 enzymes and the prediction of drug-drug interactions Drug Metab. Dispos. 2007, 35, 246– 255 DOI: 10.1124/dmd.106.012633
  51
  Mechanism-based inactivation of human cytochrome P450 enzymes and the prediction of drug-drug interactions
  Obach, R. Scott; Walsky, Robert L.; Venkatakrishnan, Karthik
  Drug Metabolism and Disposition (2007), 35 (2), 246-255CODEN: DMDSAI; ISSN:0090-9556. (American Society for Pharmacology and Experimental Therapeutics)
  The ability to use vitro inactivation kinetic parameters in scaling to in vivo drug-drug interactions (DDIs) for mechanism-based inactivators of human cytochrome P 450 (P 450) enzymes was examd. using eight human P 450-selective marker activities in pooled human liver microsomes. These data were combined with other parameters (systemic Cmax, estd. hepatic inlet Cmax, fraction unbound, in vivo P 450 enzyme degrdn. rate consts. estd. from clin. pharmacokinetic data, and fraction of the affected drug cleared by the inhibited enzyme) to predict increases in exposure to drugs, and the predictions were compared with in vivo DDIs gathered from clin. studies reported in the scientific literature. In general, the use of unbound systemic Cmax as the inactivator concn. in vivo yielded the most accurate predictions of DDI with a mean -fold error of 1.64. Abbreviated in vitro approaches to identifying mechanism-based inactivators were developed. Testing potential inactivators at a single concn. (IC25) in a 30-min preincubation with human liver microsomes in the absence and presence of NADPH followed by assessment of P 450 marker activities readily identified those compds. known to be mechanism-based inactivators and represents an approach that can be used with greater throughput. Measurement of decreases in IC50 occurring with a 30-min preincubation with liver microsomes and NADPH was also useful in identifying mechanism-based inactivators, and the IC50 measured after such a preincubation was highly correlated with the kinact/K1 ratio measured after a full characterization of inactivation. Overall, these findings support the conclusion that P 450 in vitro inactivation data are valuable in predicting clin. DDIs that can occur via this mechanism.
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52. 52
  Obach, R. S. Predicting clearance in humans from in vitro data Curr. Top. Med. Chem. 2011, 11, 334– 339 DOI: 10.2174/156802611794480873
  52
  Predicting clearance in humans from in vitro data
  Obach, R. Scott
  Current Topics in Medicinal Chemistry (Sharjah, United Arab Emirates) (2011), 11 (4), 334-339CODEN: CTMCCL; ISSN:1568-0266. (Bentham Science Publishers Ltd.)
  A review. The use of in vitro metab. in scaling to predict human clearance of new chem. entities has become a commonplace activity in the research and development of new drugs. The measurement of in vitro lability in human liver microsomes, a rich source of drug metabolizing cytochrome P 450 enzymes, has become a high throughput screen in many research organizations which is a testament to its usefulness in drug design. In this chapter, the methods used to scale in vitro intrinsic clearance data to predict in vivo clearance are described. Importantly, the numerous assumptions that are required in order to use in vitro data in this manner are laid out. These include assumptions regarding the scaling process as well as tech. aspects of the generation of the in vitro data. Finally, some other drug clearance processes that have been emerging as important are described with regard to ongoing research efforts to develop clearance prediction methods.
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53. 53
  Artursson, P.; Karlsson, J. Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells Biochem. Biophys. Res. Commun. 1991, 175, 880– 885 DOI: 10.1016/0006-291X(91)91647-U
  53
  Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells
  Artursson, P.; Karlsson, J.
  Biochemical and Biophysical Research Communications (1991), 175 (3), 880-5CODEN: BBRCA9; ISSN:0006-291X.
  Monolayers of a well differentiated human intestinal epithelial cell line, Caco-2, were used as a model to study passive drug absorption across the intestinal epithelium. Absorption rate consts. (expressed as apparent permeability coeffs.) were detd. for 20 drugs and peptides with different structural properties. The permeability coeffs. ranged from approx. 5 × 10-8 to 5 × 10-5 cm/s. A good correlation was obtained between data on oral absorption in humans and the results in the Caco-2 model. Drugs that are completely absorbed in humans had permeability coeffs. >1 × 10-6 cm/s. Drugs that are absorbed to >1% but <100% had permeability coeffs. of 0.1-1.0 × 10-6 cm/s while drugs and peptides that are absorbed to <1% had permeability coeffs. of ≤1 × 10-7 cm/s. The results indicate that Caco-2 monolayers can be used as a model for studies on intestinal drug absorption.
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54. 54
  Hoenicka, M.; Becker, E.-M.; Apeler, H.; Sirichoke, T.; Schröder, H.; Gerzer, R.; Stasch, J.-P. Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: stimulation by YC-1, nitric oxide, and carbon monoxide J. Mol. Med. (Heidelberg, Ger.) 1999, 77, 14– 23 DOI: 10.1007/s001090050292
  54
  Purified soluble guanylyl cyclase expressed in a baculovirus/Sf9 system: Stimulation by YC-1, nitric oxide, and carbon monoxide
  Hoenicka, Markus; Becker, Eva-Maria; Apeler, Heiner; Sirichoke, Tim; Schroder, Henning; Gerzer, Rupert; Stasch, Johannes-Peter
  Journal of Molecular Medicine (Berlin) (1999), 77 (1), 14-23CODEN: JMLME8; ISSN:0946-2716. (Springer-Verlag)
  Sol. guanylyl cyclase (sGC) is the main receptor for nitric oxide, a messenger mol. with multiple clin. implications. Understanding the activation of sGC is an important step for establishing new therapeutic principles. We have now overexpressed sGC in a baculovirus/Sf9 system optimized for high protein yields to facilitate spectral and kinetic studies of the activation mechanisms of this enzyme. It was expressed in a batch fermenter using a defined mixt. of viruses encoding the α1 and β1 subunits of the rat lung enzyme. The expressed enzyme was purified from the cytosolic fraction by anion exchange chromatog., hydroxyapatite chromatog., and size exclusion chromatog. By use of this new method 2.5 L culture yielded about 1 mg of apparently homogeneous sGC with a content of about one heme per heterodimer without the need of a heme reconstitution step. The enzyme did not contain stoichiometric amts. of copper. The basal activities of the purified enzyme were 153 and 1259 nmol min-1 mg-1 in the presence of Mg2+ and Mn2+, resp. The nitric oxide releasing agent 2-(N,N-diethylamino)-diazenolate-2-oxide (DEA/NO) stimulated the enzyme 160-fold with Mg2+, whereas the NO-independent activator 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) induced an increase in the activity of 101-fold at a concn. of 300 μM. The combination of DEA/NO (10 μM) and YC-1 (100 μM) elicited a dose-dependent synergistic stimulation with a max. of a 792-fold increase over the basal activity in the presence of Mg2+, resulting in a specific activity of 121 μmol min-1 mg-1. The synergistic stimulation of DEA/NO and YC-1 was attenuated by the sGC inhibitor 1H-(1,2,4)oxadiazole(4,3-a)quinoxalin-1-one (ODQ) (10 μM) by 94%. In a different exptl. setup a satd. carbon monoxide soln. in the absence of ambient oxygen or NO stimulated the enzyme 15-fold in the absence and 1260-fold in the presence of YC-1 compared to an argon control. The heme spectra of the enzyme showed a shift of the Soret peak from 432 to 399 and 424 nm in the presence of DEA/NO or carbon monoxide, resp. The heme spectra were not affected by YC-1 in the absence or in the presence of DEA/NO or of carbon monoxide, which reflects the fact that YC-1 does not interact directly with the heme group of the enzyme. In summary, this study shows that our expression/purifn. procedure is suitable for producing large amts. of highly pure sGC which contains one heme per heterodimer without a reconstitution step. The activator expts. show that in a synergistic stimulation with YC-1 sGC can be activated maximally both by nitric oxide and by carbon monoxide and that YC-1 does not directly act via heme. The described method should help to facilitate the investigation of the new therapeutic principle of NO-independent guanylyl cyclase activators.
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55. 55
  Schermuly, R. T.; Stasch, J.-P.; Pullamsetti, S. S.; Middendorff, R.; Müller, D.; Schlüter, K.-D.; Dingendorf, A.; Hackemack, S.; Kolosionek, E.; Kaulen, C.; Dumitrascu, R.; Weissmann, N.; Mittendorf, J.; Klepetko, W.; Seeger, W.; Ghofrani, H. A.; Grimminger, F. Expression and function of soluble guanylate cyclase in pulmonary arterial hypertension Eur. Respir. J. 2008, 32, 881– 891 DOI: 10.1183/09031936.00114407
  55
  Expression and function of soluble guanylate cyclase in pulmonary arterial hypertension
  Schermuly, R. T.; Stasch, J.-P.; Pullamsetti, S. S.; Middendorff, R.; Mueller, D.; Schlueter, K.-D.; Dingendorf, A.; Hackemack, S.; Kolosionek, E.; Kaulen, C.; Dumitrascu, R.; Weissmann, N.; Mittendorf, J.; Klepetko, W.; Seeger, W.; Ghofrani, H. A.; Grimminger, F.
  European Respiratory Journal (2008), 32 (4), 881-891CODEN: ERJOEI; ISSN:0903-1936. (European Respiratory Society)
  Alterations of the nitric oxide receptor, sol. guanylate cyclase (sGC) may contribute to the pathophysiol. of pulmonary arterial hypertension (PAH). In the present study, the expression of sGC in explanted lung tissue of PAH patients was studied and the effects of the sGC stimulator BAY 63-2521 on enzyme activity, and hemodynamics and vascular remodelling were investigated in two independent animal models of PAH. Strong upregulation of sGC in pulmonary arterial vessels in the idiopathic PAH lungs compared with healthy donor lungs was demonstrated by immunohistochem. Upregulation of sGC was detected, similarly to humans, in the structurally remodelled smooth muscle layer in chronic hypoxic mouse lungs and lungs from monocrotaline (MCT)-injected rats. BAY 63-2521 is a novel, orally available compd. that directly stimulates sGC and sensitizes it to its physiol. stimulator, nitric oxide. Chronic treatment of hypoxic mice and MCT-injected rats, with fully established PAH, with BAY 63-2521 (10 mg·kg-1·day-1) partially reversed the PAH, the right heart hypertrophy and the structural remodelling of the lung vasculature. Upregulation of sol. guanylate cyclase in pulmonary arterial smooth muscle cells was noted in human idiopathic pulmonary arterial hypertension lungs and lungs from animal models of pulmonary arterial hypertension. Stimulation of sol. guanylate cyclase reversed right heart hypertrophy and structural lung vascular remodelling. Sol. guanylate cyclase may thus offer a new target for therapeutic intervention in pulmonary arterial hypertension.
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56. 56
  Geschka, S.; Kretschmer, A.; Sharkovska, Y.; Evgenov, O. V.; Lawrenz, B.; Hucke, A.; Hocher, B.; Stasch, J.-P. Soluble guanylate cyclase stimulation prevents fibrotic tissue remodeling and improves survival in salt-sensitive Dahl rats PLoS One 2011, 6, e21853 DOI: 10.1371/journal.pone.0021853
  There is no corresponding record for this reference.
Supporting Information
Supporting Information
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jmedchem.7b00449.
- ¹H NMR spectra of selected compounds 3, 21, 22, and 24; CYP-inhibition results of 24 (PDF)
- Molecular formula strings (CSV)
- jm7b00449_si_001.pdf (679.41 kb)
- jm7b00449_si_002.csv (1.38 kb)
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Discovery of the Soluble Guanylate Cyclase Stimulator Vericiguat (BAY 1021189) for the Treatment of Chronic Heart Failure发现可溶性鸟苷酸环化酶激动剂维利吉乌特（BAY 1021189）用于治疗慢性心力衰竭Click to copy article link点击复制文章链接Article link copied!

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Abstract 摘要

Introduction 引言

Results and Discussion

Optimization Strategy

Chemistry

Scheme 1

Scheme 2

Scheme 3

Scheme 4

Scheme 5

Scheme 6

SAR and DMPK Optimization

Figure 1

Pharmacology

Highly Purified Recombinant sGC

Figure 2

sGC-Overexpressing Cells

Isolated Vessels and Tolerance

Langendorff-Perfused Hearts

Figure 3

Long-Term Study with L-NAME-Treated Renin Transgenic Rats

Effects on Blood Pressure and on the Heart

Figure 4

Figure 5

Effects on the Kidneys 肾脏影响

Figure 6 图 6

Effects on Mortality 死亡率影响

Figure 7 图 7

Conclusion 结论

Experimental Section 实验部分

Chemistry 化学

General Procedures 通用程序

Materials 材料

Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}(2-hydroxyethyl)carbamate (3)甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}(2-羟基乙基)甲酰胺(3)

3-Bromo-1H-pyrazolo[3,4-c]pyridazine (3c)3-溴-1H-吡唑并[3,4-c]吡啶杂氮（3c）

3-Bromo-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine (3d)3-溴-1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪（3d）

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carbonitrile (3e)1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基腈（3e）

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carboximidamide (3f)1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-羧基脒（3f）

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (3i)2-[(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪-3-基]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（3i）

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidine-4,5,6-triamine (3h)2-[1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶并[3,4-c]三胺]-4,5,6-三胺（3h）

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (4i)2-[8-(2-氟苯基咪唑[1,5-a]嘧啶-6-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（4i）

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidine-4,5,6-triamine (4j)2-[8-(2-氟苯基)咪唑并[1,5-a]嘧啶-6-基]嘧啶-4,5,6-三胺（4j）

4-(2,2,3,3-Tetrafluoropropyl)morpholine (6b)4-(2,2,3,3-四氟丙基)吗啉（6b）

4-Methyl-4-(2,2,3,3-tetrafluoropropyl)morpholin-4-ium Methanesulfonate (6c)4-甲基-4-(2,2,3,3-四氟丙基)吗啉-4-ium 甲磺酸盐（6c）

4-Methyl-4-(2,3,3-trifluoroprop-1-enyl)morpholin-4-ium Methanesulfonate (6d)4-甲基-4-(2,3,3-三氟丙-1-烯基)吗啉-4-ium 甲磺酸盐（6d）

2-Fluoro-3-(morpholin-4-yl)acrylaldehyde (6e)2-氟-3-(4-吗啉基)丙烯醛（6e）

Ethyl 5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxylate (6f)乙基 5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酸酯（6f）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxamide (6g)5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酰胺（6g）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile (6h)5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基腈（6h）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboximidamide Hydrochloride (6i)5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-羧基脒盐酸盐（6i）

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (6j)2-[(5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（6j）

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidine-4,5,6-triamine (6k)2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-4,5,6-三胺（6k）

Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidin-5-yl}carbamate Formic Acid Salt (21)甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基]嘧啶-5-基}甲酰胺甲酸盐（21）

Methyl {4,6-Diamino-2-[8-(2-fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidin-5-yl}carbamate (22)甲基{4,6-二氨基-2-[8-(2-氟苯基)咪唑[1,5-a]嘧啶-6-基]嘧啶-5-基}甲酰胺（22）

Methyl {4,6-Diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}carbamate (24)甲基{4,6-二氨基-2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}甲酰胺（24）

Biology 生物

General 通用

CYP Inhibition Assay CYP 抑制测定法

In Vitro Clearance Determinations with Rat and Human Hepatocytes体外清除度测定：大鼠和人肝细胞

Caco-2 Permeability AssayCaco-2 渗透性测定

Pharmacokinetic Parameters after Intravenous and Oral Application in Rats and Dogs**药代动力学参数在大鼠和狗体内静脉和口服给药后的研究**

Highly Purified sGC 高度纯化的 sGC

Recombinant sGC-Overexpressing Cell Line重组 sGC 高表达细胞系

Isolated Vessels and Tolerance孤立船舶与容忍

Rat Heart Langendorff Preparation大鼠心脏 Langendorff 制备

Chronic Treatment Study with L-NAME-Treated Renin Transgenic Rats慢性 L-NAME 处理肾素转基因大鼠治疗研究

Statistics 统计

Supporting Information 支持信息

Terms & Conditions 《服务条款》

Author Information 作者信息

Acknowledgment

Abbreviations Used

References

Cited By

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Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}(2-hydroxyethyl)carbamate (3)
甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}(2-羟基乙基)甲酰胺(3)

3-Bromo-1H-pyrazolo[3,4-c]pyridazine (3c)
3-溴-1H-吡唑并[3,4-c]吡啶杂氮（3c）

3-Bromo-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine (3d)
3-溴-1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪（3d）

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carbonitrile (3e)
1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基腈（3e）

1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazine-3-carboximidamide (3f)
1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-羧基脒（3f）

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (3i)
2-[(2-氟苯基)-1H-吡唑并[3,4-c]吡啶嗪-3-基]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（3i）

2-[1-(2-Fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidine-4,5,6-triamine (3h)
2-[1-(2-氟苯基)-1H-吡唑并[3,4-c]吡啶并[3,4-c]三胺]-4,5,6-三胺（3h）

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (4i)
2-[8-(2-氟苯基咪唑[1,5-a]嘧啶-6-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（4i）

2-[8-(2-Fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidine-4,5,6-triamine (4j)
2-[8-(2-氟苯基)咪唑并[1,5-a]嘧啶-6-基]嘧啶-4,5,6-三胺（4j）

4-(2,2,3,3-Tetrafluoropropyl)morpholine (6b)
4-(2,2,3,3-四氟丙基)吗啉（6b）

4-Methyl-4-(2,2,3,3-tetrafluoropropyl)morpholin-4-ium Methanesulfonate (6c)
4-甲基-4-(2,2,3,3-四氟丙基)吗啉-4-ium 甲磺酸盐（6c）

4-Methyl-4-(2,3,3-trifluoroprop-1-enyl)morpholin-4-ium Methanesulfonate (6d)
4-甲基-4-(2,3,3-三氟丙-1-烯基)吗啉-4-ium 甲磺酸盐（6d）

2-Fluoro-3-(morpholin-4-yl)acrylaldehyde (6e)
2-氟-3-(4-吗啉基)丙烯醛（6e）

Ethyl 5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxylate (6f)
乙基 5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酸酯（6f）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboxamide (6g)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-甲酰胺（6g）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carbonitrile (6h)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基腈（6h）

5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridine-3-carboximidamide Hydrochloride (6i)
5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-羧基脒盐酸盐（6i）

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]-5-[(E)-phenyldiazenyl]pyrimidine-4,6-diamine (6j)
2-[(5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基)]-5-[(E)-苯基偶氮基]嘧啶-4,6-二胺（6j）

2-[5-Fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidine-4,5,6-triamine (6k)
2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-4,5,6-三胺（6k）

Methyl {4,6-Diamino-2-[1-(2-fluorobenzyl)-1H-pyrazolo[3,4-c]pyridazin-3-yl]pyrimidin-5-yl}carbamate Formic Acid Salt (21)
甲基{4,6-二氨基-2-[(1-氟苯基)-1H-吡唑并[3,4-c]吡啶-3-基]嘧啶-5-基}甲酰胺甲酸盐（21）

Methyl {4,6-Diamino-2-[8-(2-fluorobenzyl)imidazo[1,5-a]pyrimidin-6-yl]pyrimidin-5-yl}carbamate (22)
甲基{4,6-二氨基-2-[8-(2-氟苯基)咪唑[1,5-a]嘧啶-6-基]嘧啶-5-基}甲酰胺（22）

Methyl {4,6-Diamino-2-[5-fluoro-1-(2-fluorobenzyl)-1H-pyrazolo[3,4-b]pyridin-3-yl]pyrimidin-5-yl}carbamate (24)
甲基{4,6-二氨基-2-[5-氟-1-(2-氟苯基)-1H-吡唑并[3,4-b]吡啶-3-基]嘧啶-5-基}甲酰胺（24）

In Vitro Clearance Determinations with Rat and Human Hepatocytes
体外清除度测定：大鼠和人肝细胞

Caco-2 Permeability Assay
Caco-2 渗透性测定

Pharmacokinetic Parameters after Intravenous and Oral Application in Rats and Dogs
药代动力学参数在大鼠和狗体内静脉和口服给药后的研究

Recombinant sGC-Overexpressing Cell Line
重组 sGC 高表达细胞系

Isolated Vessels and Tolerance
孤立船舶与容忍

Rat Heart Langendorff Preparation
大鼠心脏 Langendorff 制备

Chronic Treatment Study with L-NAME-Treated Renin Transgenic Rats
慢性 L-NAME 处理肾素转基因大鼠治疗研究