Soluble guanylate cyclase stimulators for the treatment of hypertension: Discovery of MK-2947

Article abstract of DOI:10.1016/j.bmcl.2020.127574

The NO-sGC-cGMP signaling pathway plays an important role in the cardiovascular system. Loss of nitric oxide tone or impaired signaling has been associated with cardiovascular diseases, such as hypertension, pulmonary hypertension and heart failure. Direct activation of sGC enzyme independent of NO represents a novel approach for modulating NO signaling with tremendous therapeutic potential. Herein, we describe the design of a structurally novel class of heme-dependent sGC stimulators containing the 3,3-dimethylpyrrolidin-2-one moiety which resulted in the identification of the potent, selective stimulator 30 (MK-2947) for the treatment of hypertension.

Full text of DOI:10.1016/j.bmcl.2020.127574

Bioorganic&MedicinalChemistryLetters30(2020)127574
Contents lists available at ScienceDirect
Bioorganic & Medicinal Chemistry Letters
journal homepage: www.elsevier.com/locate/bmcl
Soluble guanylate cyclase stimulators for the treatment of hypertension:
Discovery of MK-2947
Linda Brockuniera, John Stelmacha, Jian Guoa, Tracy Spencera, Keith Rosauera, Alka Bansalb,
Sheng-Jian Caib, Nancy Chenc, John Cummingsd, Li Huangb, Timothy Johnsone, Sonia Levesquec,
Lin Luob, Kevin Maloneyf, Joseph Metzgerc, Christopher Mortkog, Karen Ortegac, Lee-Yuh Paic,
Antonio Pereirad, Gino Saliturod, Jackie Shangd, Cherrie Shepherdc, Shiyao Sherrie Xud,
⁎
Qifeng Yangb, Jisong Cuib, Sophie Royb, Emma Parmeea, Subharekha Raghavana,
b Cardio-metabolic Disease, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
c In vivo pharmacology, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
d Pharmacokinetics, Pharmacodynamics, and Drug Metabolism, 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
e Investigative In-Vivo Safety Pharmacology, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA
f Process Research & Development, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
g Discovery Pharmaceutical Science, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
a Discovery Chemistry, Merck & Co., Inc., 2000 Galloping Hill Road, Kenilworth, NJ 07033, USA
A R T I C L E I N F O
A B S T R A C T
Keywords:
The NO-sGC-cGMP signaling pathway plays an important role in the cardiovascular system. Loss of nitric oxide
tone or impaired signaling has been associated with cardiovascular diseases, such as hypertension, pulmonary
hypertension and heart failure. Direct activation of sGC enzyme independent of NO represents a novel approach
for modulating NO signaling with tremendous therapeutic potential. Herein, we describe the design of a
structurally novel class of heme-dependent sGC stimulators containing the 3,3-dimethylpyrrolidin-2-one moiety
which resulted in the identification of the potent, selective stimulator 30 (MK-2947) for the treatment of hy-
pertension.
Soluble guanylate cyclase
Nitric oxide
Heme-dependent sGC stimulators
sGC stimulators
Hypertension
cGMP
SHR
MK-2947
NO-sGC-cGMP
Soluble guanylate cyclase (sGC), a heme containing enzyme present
in vascular smooth muscle cells and platelets, is the effector of en-
dogenous nitric oxide (NO). Binding of NO to sGC stimulates the en-
zyme to catalyze the conversion of guanosine-5′-triphosphate to cyclic
guanosine- 3′,5′-monophosphate (cGMP), an important secondary
messenger. The corresponding increase in cGMP concentration results
in several beneficial downstream physiological effects including va-
sorelaxation, inhibition of platelet aggregation and inhibition of smooth
muscle cell proliferation.1 The NO-sGC-cGMP signaling pathway plays
an important role in the regulation of the cardiovascular system. Loss of
NO-tone or impaired signaling has been associated with cardiovascular
diseases, such as hypertension, pulmonary hypertension and heart
failure.2 Several small molecule stimulators of sGC have been dis-
covered in the past several years.3 These sGC stimulators synergize with
NO and, more importantly, directly activate the native (Fe2+) state of
the enzyme independent of NO.4 Because of this dual mode of action,
heme-dependent sGC stimulators may be advantageous in preserving
and amplifying the endogenous NO signaling pathway. Riociguat
(ADEMPAS™) is a first-in-class sGC stimulator approved for treatment
of pulmonary arterial hypertension and chronic thrombo-embolic pul-
monary hypertension.5 Vericiguat demonstrated efficacy for the treat-
ment of patients with heart failure (HF) in Phase III clinical trials.6a
Praliciguat is also being evaluated for treatment of HF.6b sGC stimula-
tors are being explored for multiple indications.7 NO-independent, di-
rect activation of sGC thus represents a novel approach for modulating
NO signaling with tremendous therapeutic potential.
Given the clinical evidence and the promising literature regarding
sGC for the treatment of cardiovascular disease, we were interested in
developing a stimulator for the treatment of essential hypertension.7
Hypertension is a risk factor for coronary heart disease, myocardial
⁎ Corresponding author.
E-mail address: subha_raghavan@merck.com (S. Raghavan).
https://doi.org/10.1016/j.bmcl.2020.127574
Received 9 August 2020; Received in revised form 13 September 2020; Accepted 18 September 2020
Availableonline24September2020
0960-894X/©2020ElsevierLtd.Allrightsreserved.

L. Brockunier, et al.
Bioorganic&MedicinalChemistryLetters30(2020)127574
Fig. 1. sGC stimulator riociguat and MSD lead structures.
infarction, and stroke. It has been estimated that > 66% of patients do
not adequately control hypertension, most often due to increased sys-
tolic blood pressure (SBP).8 Drugs that form NO (e.g. nitroglycerin,
isosorbide mononitrate), which are used for treatment of angina, have
also been beneficial in treating the elderly patients with predominant or
isolated systolic hypertension.9 Despite its efficacy, however, clinical
use of nitrate therapy is limited due to the short duration of action and
the development of “nitrate tolerance”.10 We hypothesized that a non-
NO dependent sGC stimulator would provide sustained blood pressure
lowering and exert beneficial effects on end organ protection. In this
letter, we describe the discovery of structurally novel class sGC heme-
dependent stimulators resulting in the identification of the potent, se-
lective stimulator 30 (MK-2947).
Table 2
SAR of 1H-indazol-1-yl-(pyrimidin-5-yl) carbamates.
Compound
R1
R2
R3
CASA EC50 (μM)
hERG IC50 (μM)
5
Me
MeO
Cl
F
2-F Bn
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
Me
H
2.66
2.01
0.35
0.96
4.30
1.71
0.30
0.27
0.11
0.27
0.07
0.67
> 30
> 30
6.3
6
2-F Bn
7
2-F Bn
Conducting a high throughput screening campaign led to identifi-
cation of compound 1, a weak lead bearing the imidazo[1,5-a]pyridine
heterocycle motif (Fig. 1). Recognizing the potential similarity to the
1H-pyrazolo[3,4-b]pyridine scaffold in riociguat, we explored other 6–5
fused heterocyclic scaffolds, combining them with 2-fluorobenzyl sub-
stituent and the 4,6-diamino pyrimidine NeO, dimethyl carbamate
moiety present in riociguat. Thus, compounds 2–4 bearing the imidazo
[1,5-a]pyridine, imidazo[1,5-b]pyridazine and 1H-indazole scaffolds
were synthesized and tested in biological assays.11
8
2-F Bn
15.1
7.4
9
CN
H
2-F Bn
10
11
12
13
14
15
16
2-Cl Bn
10.9
7.2
H
2,3-diFBn
2,5-diFBn
2,3,5-triFBn
2,3,6-triFBn
2,3,6-triFBn
2,3,6-triFBn
H
3.4
H
28
H
> 30
3.9
Cl
Cl
> 30
The in vitro potency (Table 1) was determined by measuring cGMP
production in CHO cells over-expressing the human sGC α1/β1 isoform
(CASA assay). In vivo efficacy was evaluated in conscious spontaneously
hypertensive rat (SHR) model implanted with telemetry device that
enabled continuous monitoring of hemodynamic parameters.11 Systolic
blood pressure (SBP) decreases for compounds 2–4 measured 4-hours
post oral administration are summarized in Table 1. Compound 4,
containing a 1H-indazole scaffold with carbon-linked 2-fluoro benzyl
substituent, was the most potent (CASA EC50 = 0.3 μM) and exhibited a
good off-target profile, particularly against hERG. Hence, further opti-
mization was focused on this scaffold with an eye toward improving
potency and pharmacokinetic properties. With this goal in mind,
compounds 5–16 (Table 2) were synthesized to explore the SAR of
substituents at the C-5 position of the 1H-indazole core (R1), benzyl
group (R2) and the carbamate nitrogen (R3).
with chloro and fluoro analogs (compound 7 and 8), having minimal
effect on sGC potency, while introduction of alkyl groups resulted in
decreased activity compared to compound 4. Exploration of benzyl
group SAR (R2) led to the identification of di-fluoro and trifluoro ana-
logs with similar or enhanced potency relative to compound 4 (Table 2,
compounds 11–15). Compound 15, featuring a 2,3,6-trifluorobenzyl
group, showed
a
significant improvement in potency (CASA
EC50 = 0.07 μM), albeit with decreased hERG selectivity relative to
lead 4. In the SHR model, dosing of compound 15 led to a decrease in
SBP by 60 mmHg at 3 mg/kg dose, with the BP effects lasting for over
24 h (-40 mmHg at 24 h). The duration of effect was surprising given
that the half-life of compound 15 in a rat PK study was ~3.6 h
(Table 3). On further examination, it was discovered compound 15
underwent in vivo N-demethylation at R3 to afford compound 16, a
The C-5 position of 1H-indazole (R1) was sensitive to substitution
Table 3
SD Rat PK properties of compounds 8, 14, 15, and 16.
Table 1
A comparison of the activity of compounds 1–4.
8
14
15
16e
Compound CASAa EC50
SBP SH ratsb
Rat PKc T1/2
hERG IC50
(μM)
Clpa (ml/min/kg)
AUCnormb(μM.h kg/mg)
t1/2c(h)
1.8
4.6
1.5
22
1.6
15.3
2.2
68
0.9
18
~0.24
25.2
~9
(μM)
(mmHg)
(h), %F
3.6
51
1
2
3
4
4.6
20.4
1.0
n.t.
n.t.
n.t.
Forald(%)
37
−20
−30
−55
0.9,15
0.6,4
0.9,32
0.40
16.1
> 30
a
Total plasma clearance; bAUCnorm dose-normalized area under the plasma
concentration vs. time curve following oral dosing; cterminal half-life; doral
bioavailability; edue to the long half-life, the duration of the study was in-
sufficient to fully define the PK parameters.
0.31
a
Potency measured in CHO cell (n = 1), bSBP (systolic blood pressure re-
duction 4 h post oral dose of 30 mg/kg), drat IV (1 mg/kg) and PO (2 mg/kg).
2

L. Brockunier, et al.
Bioorganic&MedicinalChemistryLetters30(2020)127574
Table 4
Table 6
SAR of 7,9-dihydro-purin-8-one and 5,7-dihydro-6H-pyrrolo[2,3-d]pyrimidin-
Pharmacokinetic Properties of compound 30 in in rat, dog and rhesus monkey.a
6-one analogs.
Species
Dose (mg/kg)
CL (ml/
AUCNpo (μM.h.kg/
t1/2 (h) Fpo (%)
(IV/PO)
min/kg)
mg)
Rat
dog
1/2
29
39
34
0.98
0.23
0.74
2.6
1.3
1.4
80
21
63
1/10
monkey 0.5/2
a
Pharmacokinetic parameters were obtained following oral administration
in DMA:PEG400:water (10:45:45; v/v/v) to rats, imwitor742/tween 80 (50:50;
v/v) to dogs and monkeys.
Table 7
Changes in hemodynamic parameters and plasma levels in SHRs induced by
compound 30.
Dosage (mg/kg)
0.3
1.0
3.0
Δmean BP (mmHg)
Δsystolic BP (mmHg)
Δdiastolic BP (mmHg)
Δheart rate (bpm)
Plasma level (µM) 4 h
post-dose
12
3
3
25
4
5
5
9
36
3
14
28
38
3
8
3
20
30
3
Compound
R1
R3
X
CASA EC50 (μM)
hERG IC50 (μM)
17
8
58
86
12
0.036
0.006
0.145
0.036
0.392
0.089
17
18
19
20
21
H
H
H
Cl
H
H
NH
3.3
> 15
> 30
> 30
> 30
> 30
Me
Et
Me
–
NMe
NEt
0.12
0.02
0.14
0.16
NMe
CH2
circulating metabolite with a long rat half-life (> 9h, Table 3). Based on
this result, we hypothesized that 16 could be contributing to the ex-
tended SBP lowering effect observed. Independent synthesis of com-
pound 16 and evaluation in the SHR model at a 3 mg/kg dose led to a
decrease in SBP by 30 mmHg that was sustained for 24 h. Attenuation
of the in vivo efficacy of des-methyl analog 16 compared to compound
15 was expected given the potency difference between the two com-
pounds.
Table 5
SAR of gem-dimethyl lactams.
To avoid the presence of an active circulating metabolite with a long
duration of action, we explored ways of increasing metabolic stability
of the N-methyl carbamate moiety on the pyrimidine ring while
maintaining the sGC enzyme potency and in vivo efficacy of compound
15 and improving the hERG selectivity. One of the first design elements
explored was cyclization of the carbamate moiety with the pendant
amino group to form the N-alkyl-7,9-dihydro-purin-8-one type struc-
tures exemplified in Table 4.
Compound R1 R2
CASA EC50
SBP SHRa
(mmHg)
hERG
(nM)
IC50 (μM)
22
23
24
25
26
27
28
29
30
31
H
2-F Bn
3
−40
n.t.
4.9
Cl 2-F Bn
18
2
1.5
H
2,3,6-triFBn
n.t.
2.2
Compounds 18–20, with C-7 methyl or ethyl substituents, were
potent in vitro and exhibited minimal hERG activity. Compound 18 was
tested in vivo and had a short rat half-life and poor oral bioavailability
(t1/2 = 0.3 h; %F = 3) and showed no in vivo efficacy in SHRs at
10 mg/kg. Reasoning that the poor PK profile of analog 18 could again
be attributed to N-demethylation, we explored replacement of the ni-
trogen atom with a carbon atom to eliminate the possibility of com-
petitive de-methylation to afford 5,7-dihydro-6H-pyrrolo[2,3-d]pyr-
imidin-6-one type analogs such as compound 21.
Cl 2,3,6-triFBn
Cl CH2 CH2CH3
7.3
546
47
2.3
−80
n.t.
1.4
9.6
Cl CH2CH2CH2CH3
−40
n.t.
12.3
9.0
Cl CH2CH2CH2CH2CH3 90
H
CH2CH2CF3
882
87
n.t.
> 30
> 30
4.1
Cl CH2CH2CF3
28
−40
−40
Cl CH2CH2CH2CF3
117
a
Maximum peak decreases of SBP.
Scheme 1. Synthesis of 30. Reagents and conditions: (a) t-BuOK (1.0 eq.), THF, 50 °C, 18 h (100%); (b) Aminoguanidine HCl (1.5 eq.), BF3OEt2 (2.0 eq.), MeOH
100 °C, 1.5 h, (quantitative, 100%); (c) 34 (2.0 eq.), t-BuOK (1.0 eq.), n-butanol, 120 °C, 1.5 h (99%); (d) CuI (0.2 eq.), N,N' dimethylethylenediamine (2.0 eq.), DMF,
rt, 0.5 h (41%).
3

L. Brockunier, et al.
Bioorganic&MedicinalChemistryLetters30(2020)127574
Compound 21 exhibited similar potency to analog 18 and was ef-
ficacious in vivo (decreased SBP 20 mmHg; 3 mg/kg); however, it ex-
hibited very high clearance and a short half-life in rats (Cl = 95 mL/
min/kg, t1/2 = 0.5 h, %F = 6). Furthermore, compound 21 was un-
stable, undergoing oxidative degradation upon standing at room tem-
perature to form the corresponding pyrrolidine-2,3-dione. Gem-di-
methyl substitution on the lactam methylene was pursued to mitigate
the chemical instability of the parent 21. Hence, compounds 22–25,
which combined the potency-enhancing R1 and R2 groups identified
from the previous SAR efforts, were synthesized (Table 5).
decrease in mean SBP (Table 7). The corresponding plasma levels were
determined in a parallel pharmacokinetic study. Administration of
compound 30 (0.3 and 1 mg/kg/day, PO) daily to SHR for 14 con-
secutive days evoked consistent decreases in SBP that were similar in
magnitude to those determined in single dose studies (~10–15 mmHg
at 0.3 mg/kg and ~20–30 mmHg at 1 mg/kg). Heart rate changes were
attenuated, indicating the presence of reflex tachycardia.
In summary, we have discovered compound 30, a novel potent,
selective sGC stimulator, also known as MK-2947. To address the issue
of N-demethylation of the original lead 15, medicinal chemistry efforts
focused on designing cyclic analogs that were not susceptible to oxi-
dation or demethylation. These efforts led to discovery of the 3,3-di-
methylpyrrolidin-2-one group, which afforded several potent sGC sti-
mulators that robustly lowered BP in SHR model. Compound 30 was
selected as a clinical candidate based on its potency, off-target se-
lectivity, pharmacokinetic and favorable pharmaco-dynamic profile in
preclinical species. Further studies on MK-2947 are pending and will be
reported in due course.
A key breakthrough was the discovery of analog 25, which com-
bined the choro-substituent at R1 with the 2,3,6-tri-fluoro benzyl sub-
stituent at R2 identified from previous SAR findings. Compound 25 was
extremely potent both in vitro (CASA EC50 = 7 nM) and in vivo (peak
SBP decrease of 80 mmHg at 3 mg/kg), with a favorable rat pharma-
cokinetic profile (T1/2 = 4 h, %F = 53). An unfortunate liability with
analog 25 was the loss in hERG selectivity. Consequently, replacement
of the fluorobenzyl groups at R2 with aliphatic groups (compounds
26–31) was explored to mitigate hERG activity. Compound 27, bearing
a butyl group at R2, exhibited the highest potency, followed by the
pentyl analog 28. These compounds likewise showed steady improve-
ment in hERG selectivity. Replacing the R2 terminal methyl groups on
compounds 26–28 with a trifluoromethyl group afforded compounds
29–31. Interestingly, compound 29 had no hERG activity
(IC50 > 30 μM) but lost 10-fold potency against the sGC enzyme
(CASA EC50 = 882 nM) when the R1 substituent was a hydrogen. This
loss in potency was restored by installing the choro-group at R1 (com-
pound 30 CASA IC50 = 87 nM). Increasing the R2 chain length to a
trifluoro-butyl group resulted in an erosion in hERG selectivity.
Compound 30 differentiated from the rest of the series due to its sGC
potency and hERG selectivity and was ultimately chosen for further
profiling as a potential clinical candidate.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.bmcl.2020.127574.
References
In order to provide sufficient quantities of compound 30 for further
evaluation, a five-step synthesis of compound 30 was accomplished
(Scheme 1). Methyl 3,3-dicyano-2,2-dimethyl propanoate (34) was
obtained by treating malononitrile (32) with potassium tert-butoxide
and methyl-2-bromo isobutyrate (33) in THF. Treatment of the ketone
3511 with aminoguanidine hydorochloride in the presence of boron
trifluoro etherate in methanol afforded the aminoguanidine hydrazone
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of penultimate 37 resulted in the indazole ring formation to afford the
desired compound 30 in 41% yield.
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