Pyrazolo[3,4-d]pyrimidinone compounds for the treatment of chronic heart failure

Abstract

This application relates to a series of novel compounds that are selective inhibitors of phosphodiesterase type 9 ("PDE9") for the treatment of chronic heart failure. More particularly, the application relates to pyrazolo[3,4-d]pyrimidinone compounds for use in the treatment and prevention of chronic heart failure.

Description

Attorney Docket No.: CDPH-003 / 001WO 341287-2404 PYRAZOLO[3,4-D]PYRIMIDINONE COMPOUNDS FOR THE TREATMENT OF CHRONIC HEART FAILURE RELATED APPLICATIONS

[0001] The present application claims priority to, and the benefit of, US Provisional Application No. 63 / 533,961 filed August 22, 2023, and US Provisional Application No. 63 / 643,495 filed May 7, 2024, the contents of each of which are incorporated herein by reference in their entirety. BACKGROUND

[0002] Approximately 6 million adults in the United States have heart failure, and heart failure is the most frequent cause of hospitalization in those older than 65 years. Heart failure costs the United States about $30.7 billion per year with 50% of patients dying within 5 years. New therapeutics that reduce morbidity and mortality in heart failure are needed.

[0003] Recently, evidence has emerged supporting that PDE9A plays an important role in the biology of heart failure and is upregulated in cardiac myocytes of patients with heart failure, dampening signaling through the beneficial natriuretic peptide receptor / cGMP / protein kinase GI (PKGI) signaling pathway.

[0004] The cyclic guanosine monophosphate (cGMP) / protein kinase GI (PKGI) signaling pathway plays a critical role in cardiovascular pathophysiology. For example, PKGI activation inhibits cardiac hypertrophy and remodeling. In the cardiomyocyte, signaling through the cGMP / PKGI pathway is achieved via four hormones; nitric oxide (NO) and three natriuretic peptides (atrial natriuretic peptide [ANP], b-type natriuretic peptide [BNP], and c- type natriuretic peptide [CNP]). NO acts through the soluble guanylate cyclase (sGC) receptor located in the cytoplasm, CNP acts through the guanylate cyclase type B receptor (GC-B), and both ANP and BNP act through the guanylate cyclase type A receptor (GC-A) located in the cell membrane. Stimulation of guanylate cyclase results in the conversion of guanosine triphosphate to cGMP.

[0005] cGMP is a second messenger molecule that transduces NO- and natriuretic-peptide- coupled signaling. cGMP binds to and activates protein kinase G leading to phosphorylation changes of protein kinase G substrates in the myocardium. cGMP generally has a protective function on the heart, attenuating adverse remodeling. The breakdown or hydrolysis of cGMPAttorney Docket No.: CDPH-003 / 001WO 341287-2404 in the cardiovascular system can occur under the action of two enzymes, PDE-5 and PDE-9. Inhibition of PDE-5 and / or PDE-9 could augment cGMP levels by blocking its degradation. Inhibitors of PDE9 have been described, see, e.g., U.S. Patent No. 7,964,607, which is incorporated herein by reference in its entirety.

[0006] Both heart failure with reduced ejection fraction (HFrEF) and heart failure with preserved ejection fraction (HFpEF) are major health problems with a significant impact on morbidity, mortality, quality of life, and healthcare costs. They are leading global causes of hospitalizations, and despite treatment advances, 5-year mortality rates remain approximately 50% within 5 years of first diagnosis. New therapeutics, such as PDE-9 inhibitors, that reduce morbidity and mortality in heart failure (HF) are needed. SUMMARY

[0007] In one aspect, provided herein is a method of treating chronic heart failure in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, wherein the compound has the structure of Formula (I),or a pharmaceutically acceptable salt thereof, wherein: R is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C3- C8)cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, each of which optionally may be substituted with one to three substituents, the substituents being independently selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, halo, and (C1-C4)haloalkyl, R1is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C2-C4)alkenyl, (C2- C4)alkynyl, (C1-C4)haloalkyl, and cyclopropyl; R2is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1- C6)haloalkyl, heteroaryl selected from the group consisting of pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, and ER5, wherein the heteroaryl optionally may be substituted with one to three substituents independently selected from the group consisting of (C1- C4)alkyl and (C1-C4)haloalkyl;Attorney Docket No.: CDPH-003 / 001WO 341287-2404 R3is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C2-C4)alkenyl, (C2- C4)alkynyl, (C3-C6)cycloalkyl, and (C1-C4)haloalkyl; E is selected from the group consisting of —CH2—, —CH2CH2—, —CH2CH2CH2—, and — C(O)—; and R5is selected from the group consisting of (C3-C8)cycloalkyl, heterocycloalkyl, aryl, aryloxy, and heteroaryl, any of which optionally may be substituted with one to three substituents, such substituents being independently selected from the group consisting of (C1-C4)alkyl, (C2-C4)alkenyl, (C2-C4)alkynyl, (C1-C4)hydroxyalkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1-C4)haloalkoxy, (C3-C8)cycloalkyl, halo, cyano, phenyl, morpholinyl, (C1- C4)alkylamino, pyrazolyl, triazolyl, and imidazolyl.

[0008] In another aspect, provided herein is a method of treating chronic heart failure in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound where the compound is 6-((3S,4S)-4-methyl-1-(pyrimidin-2- ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one (“Compound A”) or a pharmaceutically acceptable salt thereof.

[0009] In some embodiments, the method comprises administering 5 mg to 50 mg of the compound to the subject. In some embodiments, the method comprises administering 5 mg to 25 mg of the compound to the subject. In some embodiments, the method comprises administering 50 mg of the compound to the subject. In some embodiments, the method comprises administering the compound twice daily. In some embodiments, the method comprises administering the compound every 12 hours. In some embodiments, the method comprises administering the compound for at least two weeks. In some embodiments, the method comprises administering the compound for at least 10 weeks. In some embodiments, the method comprises administering the compound for the rest of the subject’s life. In some embodiments, the method comprises administering the compound of Formula (I) orally.

[0010] In some embodiments, the chronic heart failure is chronic heart failure with reduced ejection fraction. In some embodiments, the chronic heart failure is chronic heart failure with preserved ejection fraction. In some embodiments, the chronic heart failure is chronic heart failure with midrange ejection fraction. In some embodiments, the chronic heart failure is chronic heart failure with super ejection fraction.

[0011] In some embodiments, the subject has plasma or serum NT-proBNP levels of at least 600 pg / ml prior to the administration.

[0012] In some embodiments, the method results in a reduction in N-terminal pro b-type natriuretic peptide (NT-proBNP) levels of about 30% as measured in the plasma or serum ofAttorney Docket No.: CDPH-003 / 001WO 341287-2404 the subject. In some embodiments, the method results in a reduction in b-type natriuretic peptide (BNP) levels of about 30% as measured in the plasma or serum of the subject. In some embodiments, the method results in an increase in cGMP levels of about 30% as measured in the plasma or serum of the subject. In some embodiments, the method results in an increase in cGMP levels of about 30% as measured in the urine of the subject. In some embodiments, the method results in an increase in the ratio of plasma or cGMP to plasma NT-proBNP levels of about 30%. In some embodiments, the method results in an increase in the ratio of plasma cGMP to plasma BNP levels of about 30%. In some embodiments, the method results in an increase in the Kansas City Cardiomyopathy Questionnaire-23 (KCCQ- 23) score of at least 5 points.

[0013] In another aspect, provided herein is a method of treating chronic heart failure in a subject in need thereof, the method comprising orally administering to the subject 25 mg of 6-((3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (“Compound A”), or a pharmaceutically acceptable salt thereof, every 12 hours. BRIEF DESCRIPTION OF DRAWINGS

[0014] Fig. 1 is a schematic of Part A of the study described in Example 3. Abbreviations: D = Day; q12h = every 12 hours; V = Visit.

[0015] Fig. 2 is a schematic of Part B of the study described in Example 3. Abbreviations: D = Day; V = Visit.

[0016] Fig. 3 is a graph of geometric mean (ng / mL) of urinary cGMP as a function of time measured at day 0 (baseline), 1, 14, and 28 for the placebo and Compound A.

[0017] Fig. 4 is two graphs showing the mean percent change from the baseline for plasma cGMP as a function of time measured at day 1 and week 4 of administration.

[0018] Fig. 5 is two graphs showing the time-matched mean percent change from baseline in plasma cGMP as a function of time measured at week 4 for Entresto-treated and Entresto- naïve patients.

[0019] Fig. 6 is a graph showing the change in the geometric mean (95% CI) of NT-proBNP (pg / mL) measured at day 0, 14, 28, and 84 for patients treated with the placebo and Compound A.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0020] Fig. 7 is a graph showing the mean change in KCCQ scores (i.e., Clinical Summary Score (CSS), Overall Summary Score (OSS), and Total Summary Score (TSS)) between the placebo and Compound A groups at week 12.

[0021] Fig. 8 is three graphs showing the percentage of patients with >5-, >10-, and >20- point increases in magnitude to the OSS and CSS KCCQ summary scores between the placebo and Compound A groups at week 12. DETAILED DESCRIPTION

[0022] Provided herein are methods of treating chronic heart failure using selective PDE9 inhibitors, as well as unit dosage forms and pharmaceutical compositions. Compounds

[0023] The present application comprises selective PDE9 inhibitors of Formula (I),and pharmaceutically acceptable salts thereof, wherein: R is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C3- C8)cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, each of which optionally may be substituted with one to three substituents, the substituents being independently selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, halo, and (C1-C4)haloalkyl. R1is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C2-C4)alkenyl, (C2- C4)alkynyl, (C1-C4)haloalkyl, and cyclopropyl; R2is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1- C6)haloalkyl, heteroaryl selected from the group consisting of pyridinyl, pyridazinyl,pyrimidinyl, and pyrazinyl, and ER5, wherein the heteroaryl optionally may be substituted with one to three substituents independently selected from the group consisting of (C1- C4)alkyl and (C1-C4)haloalkyl; R3is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C1-C4)alkenyl, (C2- C4)alkynyl, (C3-C6)cycloalkyl, and (C1-C4)haloalkyl; E is selected from the group consisting of —CH2—, —CH2CH2—, —CH2CH2CH2—, and — C(O)—; andAttorney Docket No.: CDPH-003 / 001WO 341287-2404 R5is selected from the group consisting of (C3-C8)cycloalkyl, heterocycloalkyl, aryl, aryloxy, and heteroaryl, any of which optionally may be substituted with one to three substituents, such substituents being independently selected from the group consisting of (C1-C4)alkyl, (C2-C4)alkenyl, (C2-C4)alkynyl, (C1-C4)hydroxyalkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1-C4)haloalkoxy, (C3-C8)cycloalkyl, halo, cyano, phenyl, morpholinyl, (C1- C4)alkylamino, pyrazolyl, triazolyl, and imidazolyl.

[0024] Preferably, R is selected from the group consisting of ethyl, isopropyl, trifluoroethyl, cyclobutyl, cyclopentyl, difluorocyclohexyl, methoxyphenyl, and tetrahydro-2H-pyran-4-yl; R1is hydrogen or methyl; R2is methyl, trifluoroethyl, trifluorobutyl, pyrimidinyl, trifluoromethylpyrimidinyl, or ER5; R3is methyl, ethyl, isopropyl, trifluoromethyl, trifluoroethyl, or cyclopropyl; E is —CH2— or —C(O)—; and R5is selected from the group consisting of substituted or unsubstituted cyclopentyl, morpholinyl, phenyl, naphthyl, benzyloxy, pyrimidinyl, pyridinyl, quinolinyl, quinoxalinyl, pyrazinyl, pyrazolyl, benzimidazolyl, cinnolinyl, naphthydrinyl, pyrido[2,3-b]pyrazinyl, imidazo[4,5-c]pyridinyl, benzothiadiazolyl, tetrahydropyrazolo[1,5-a]pyridinyl, dihydrobenzodioxinyl, imidazolyl, dihydrobenzofuranyl, triazolyl, oxazolyl, isoxazolyl, benzodioxinyl, thiazolyl, imidazo[1,2- a]pyridinyl, tetrahydrobenzothiazolyl, dihydrobenzoxazinyl, tetrahydropyranyl, tetrahydropyrazolo[1,5-a]azepinyl, and dihydropyrrolo[1,2-b]pyrazolyl.

[0025] More preferably, R is selected from the group consisting of isopropyl, cyclobutyl, cyclopentyl, and tetrahydro-2H-pyranyl; R1is hydrogen; R2is ER5; R3is methyl or ethyl; E is —CH2—; and R5is selected from the group consisting of phenyl, pyrimidin-2-yl, pyridin-2- yl, pyrazin-2-yl, and 5-methylpyrazin-2-yl.

[0026] In other preferred embodiments, the compound is:

[0027] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(2-methoxyphenyl)-1,5-dihydro- 4H-pyrazolo [3,4-d]pyrimidin-4-one

[0028] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H- pyrazolo [3,4-d]pyrimidin-4-one;

[0029] 6-[(3S,4S)-1-benzyl-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0030] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-pyrimidin-2-ylpyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0031] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[4-(trifluoromethyl)pyrimidin-2- yl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0032] 6-[(3,4-trans)-1-benzoyl-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H- pyrazolo [3,4-d]pyrimidin-4-one;

[0033] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(pyridin-3-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0034] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[3-(trifluoromethyl)benzyl]pyrrolidin-3- yl}1,5-dihydro-4H -pyrazolo [3,4-d]pyrimidin-4-one;

[0035] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinolin-2-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0036] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinolin-4-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0037] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-{[6-(trifluoromethyl)pyridin-3- yl]methyl}pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0038] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinoxalin-2-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0039] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0040] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(pyrimidin-5-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0041] 1-cyclopentyl-6-[(3,4-trans)-1,4-dimethylpyrrolidin-3-yl]-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0042] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(2,2,2-trifluoroethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0043] 1-cyclopentyl-6-(3,4-trans)-4-methyl-1-[(2-methylpyridin-3-yl)methyl]pyrrolidin-3- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0044] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinolin-8-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0045] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(quinolin-3-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0046] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(6-methylpyridin-3-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0047] 6-[(3,4-trans)-1-benzyl-4-isopropylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0048] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-cyclopentyl-3-methyl-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0049] 1-cyclopentyl-6-[(3S,4S)-4-methyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0050] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(2-phenylethyl)pyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0051] 1-cyclopentyl-6-{(3,4-trans)-1-[(6-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0052] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(pyridin-2-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0053] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(3-methylpyridin-2-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0054] 6-[(3,4-trans)-1-benzyl-4-ethylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0055] 6-[(3,4-trans)-1-benzyl-4-cyclopropylpyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0056] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-isopropyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0057] 6-[(3S,4S)-1-benzyl-4-methylpyrrolidin-3-yl]-1-isopropyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0058] 1-isopropyl-6-[(3,4-trans)-4-methyl-1-(quinolin-2-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0059] 1-isopropyl-6-[(3,4-trans)-4-methyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0060] 1-isopropyl-6-[(3,4-trans)-4-methyl-1-(quinolin-3-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0061] 6-[(3,4-trans)-1-benzyl-4-(trifluoromethyl)pyrrolidin-3-yl]-1-cyclopentyl-1,5-dihydro- 4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0062] 1-isopropyl-6-[(3S,4S)-4-methyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0063] 1-isopropyl-6-[(3S,4S)-4-methyl-1-(quinolin-3-ylmethyl)pyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0064] 1-cyclopentyl-6-{(3S,4S)-4-methyl-1-[(5-methylpyrazin-2-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0065] 6-[(3,4-trans)-1-benzyl-4-ethylpyrrolidin-3-yl]-1-isopropyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0066] 6-[(3S,4S)-1-benzyl-4-ethylpyrrolidin-3-yl]-1-isopropyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0067] 1-isopropyl-6-{(3S,4S)-4-methyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0068] 6-(1-benzylpyrrolidin-3-yl)-1-cyclopentyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin- 4-one;

[0069] 1-isopropyl-6-(3S,4S)-1-[(6-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin-3-yl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0070] 6-[(3,4-trans)-4-ethyl-1-(quinolin-3-ylmethyl)pyrrolidin-3-yl]-1-isopropyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0071] 6-{(3,4-trans)-4-ethyl-1-[(6-methoxypyridin-3-yl)methyl]pyrrolidin-3-yl}-1- isopropyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0072] 6-[(3,4-trans)-4-ethyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]-1-isopropyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0073] 1-cyclopentyl-6-{(3S,4S)-1-[(1,3-dimethyl-1H-pyrazol-5-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0074] 1-cyclopentyl-6-[(3S,4S)-4-methyl-1-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3- ylmethyl) pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0075] 1-cyclopentyl-6-{(3S,4S)-4-methyl-1-[(1-methyl-1H-benzimidazol-2-yl) methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0076] 1-isopropyl-6-{(3S,4S)-4-methyl-1-[(5-methylpyrazin-2-yl)methyl]pyrrolidin-3-yl}- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0077] 6-[(3S,4S)-1-(cinnolin-3-ylmethyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0078] 1-cyclopentyl-6-[(3,4-trans)-1-(quinoxalin-6-ylmethyl)-4-(trifluoromethyl)pyrrolidin- 3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0079] 1-cyclopentyl-6-{(3S,4S)-4-methyl-1-[(2-methylpyrimidin-4-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0080] 1-cyclopentyl-6-[(3S,4S)-1-{[2-(dimethylamino)pyrimidin-4-yl]methyl}-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0081] 1-cyclopentyl-6-{(3,4-trans)-4-cyclopropyl-1-[(5-methylpyrazin-2- yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0082] 1-cyclopentyl-6-[(3,4-trans)-4-cyclopropyl-1-(quinoxalin-6-ylmethyl)pyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo [3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0083] 1-cyclopentyl-6-[(3,4-trans)-4-cyclopropyl-1-methylpyrrolidin-3-yl]-1,5-dihydro-4H- pyrazolo [3,4-d]pyrimidin-4-one;

[0084] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-ethyl-1,5-dihydro-4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0085] 6-{(3,4-trans)-4-ethyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin-3-yl}-1- isopropyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0086] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0087] 6-[(3S,4S)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0088] 6-[(3,4-trans)-1-benzyl-4-(2,2,2-trifluoroethyl)pyrrolidin-3-yl]-1-cyclopentyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0089] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(4,4-difluorocyclohexyl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0090] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(2,2,2-trifluoroethyl)-1,5-dihydro- 4H-pyrazolo [3,4-d]pyrimidin-4-one;

[0091] 1-isopropyl-6-[(3S,4S)-4-methyl-1-(1,5-naphthyridin-4-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0092] 1-isopropyl-6-[(3S,4S)-4-methyl-1-(1,8-naphthyridin-4-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0093] 1-isopropyl-6-[3S,4S)-4-methyl-1-(quinolin-4-ylmethyl)pyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0094] 1-isopropyl-6-[(3S,4S)-4-methyl-1-(pyrido[2,3-b]pyrazin-8-ylmethyl)pyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0095] 1-isopropyl-6-{(3,4-trans)-1-[(6-methoxy-1,5-naphthyridin-4-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0096] 6-{(3,4-trans)-1-[(8-fluoroquinolin-2-yl)methyl]-4-methylpyrrolidin-3-yl}-1- isopropyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0097] 1-isopropyl-6-{(v)-1-[(6-methoxyquinolin-4-yl)methyl]-4-methylpyrrolidin-3-yl}-1,5- dihydro-4H-pyrazolo[3.4-d]pyrimidin-4-one;

[0098] 6-[(3,4-trans)-1-benzyl-4-methylpyrrolidin-3-yl]-1-cyclobutyl-1,5-dihydro-4H- pyrazolo[3,4-d]pyrimidin-4-one;

[0099] 6-{(3S,4S)-4-methyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0100] 6-{(3,4-trans)-4-ethyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0101] 6-{(3S,4S)-4-methyl-1-[(5-methylpyrazin-2-yl)methyl]pyrrolidin-3-yl}-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0102] 6-{(3S,4S)-1-[(6-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0103] 6-[(3S,4S )-4-methyl-1-(quinolin-3-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0104] 6-{(3S,4S)-4-methyl-1-[(2-methylpyrimidin-4-yl)methyl]pyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0105] 6-{(3S,4S)-4-methyl-1-[(6-methylpyridin-3-yl)methyl]pyrrolidin-3-yl}-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0106] 6-{(3S,4S)-4-methyl-1-}([6-(trifluoromethyl)pyridin-3-yl]methyl}pyrrolidin-3-yl]-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0107] 6-{(3S,4S)-4-methyl-1-[(1-methyl-1H-imidazo[4,5-c]pyridin-2-yl)methyl]pyrrolidin- 3-yl}-1-(tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0108] 6-{(3S,4S)-1-[(1,3-dimethyl-1H-pyrazol-5-yl)methyl]-4-methylpyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0109] 1-cyclobutyl-6-{(3,4-trans)-4-methyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0110] 6-[(3S,4S)-1-(2,1,3-benzothiadiazol-5-ylmethyl)-4-methylpyrrolidin-3-yl]-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0111] 6-[(3S,4S)-4-methyl-1-(quinoxalin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0112] 6-[(3S,4S)-4-methyl-1-(quinolin-4-ylmethyl)pyrrolidin-3-yl]-1-tetrahydro-2H-pyran- 4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0113] 6-[(3S,4S)-4-methyl-1-(pyridin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H-pyran- 4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0114] 6-[(3S,4S)-1-benzyl-4-methylpyrrolidin-3-yl]-3-methyl-1-(tetrahydro-2H-pyran-4-yl)- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0115] 6-[(3S,4S)-1-(3-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0116] 6-[(3S,4S)-1-(35-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0117] 6-{(3S,4S)-4-methyl-1-[4-(trifluoromethyl)benzyl]pyrrolidin-3-yl}-1-(tetrahydro-2H- pyran-4-y)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0118] 6-[(3,4-trans)-1-benzyl-4-ethylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0119] 6-[(3S,4S)-1-benzyl-4-ethylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0120] 3-methyl-6-[(3S,4S)-4-methyl-1-(pyridin-3-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0121] 3-methyl-6-{(3S,4S)-4-methyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin-3-yl}-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0122] 6-{(3S,4S)-1-[(6-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin-3-yl}-3-methyl-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0123] 6-{(3S,4S)-4-methyl-1-[(6-methylpyridin-2-yl)methyl]pyrrolidin-3-yl}-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0124] 6-[(3S,4S)-1-(4-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0125] 6-[(3S,4S)-1-(2-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4-yl)- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0126] 6-{(3S,4S)-4-methyl-1-[2-(trifluoromethyl)benzyl]pyrrolidin-3-yl}-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0127] 6-[(3S,4S)-1-(2,4-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran- 4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0128] 6-[(3S,4S)-1-(4-methoxybenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0129] 6-[(3S,4S)-1-benzyl-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-thiopyran-4-yl)-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0130] 6-[(3S,4S)-1-(2-methoxybenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0131] 6-[(3S,4S)-1-(3-methoxybenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0132] 6-{(3S,4S)-4-methyl-1-[3-(trifluoromethyl)benzyl]pyrrolidin-3-yl}-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0133] 6-[(3S,4S)-1-(26-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0134] 6-{(3S,4S)-4-ethyl-1-[(5-methylpyrazin-2-yl)methyl]pyrrolidin-3-yl}-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0135] 6-{(3S,4S)-4-ethyl-1-[(6-methoxypyridin-3-yl)methyl]pyrrolidin-3-yl}-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0136] 6-[(3S,4S)-4-ethyl-1-(pyridin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H-pyran-4- yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0137] 6-[(3S,4S)-4-ethyl-1-(quinoxalin-2-ylcarbonyl)pyrrolidin-3-yl]-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0138] 6-[(3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl]-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0139] 2-({(3S,4S)-3-ethyl-4-[4-oxo-1-(tetrahydro-2H-pyran-4-yl)-45-dihydro-1H- pyrazolo[3,4-d]pyrimidin-6-yl]pyrrolidin-1-yl}methyl)benzonitrile;

[0140] 3-({(3S,4S)-3-ethyl-4-[4-oxo-1-(tetrahydro-2H-pyran-4-yl)-45-dihydro-1H- pyrazolo[3,4-d]pyrimidin-6-yl]pyrrolidin-1-yl}methyl)benzonitrile;

[0141] 4-({(3S,4S)-3-ethyl-4-[4-oxo-1-(tetrahydro-2H-pyran-4-yl)-45-dihydro-1H- pyrazolo[3,4-d]pyrimidin-6-yl]pyrrolidin-1-yl}methyl)benzonitrile;

[0142] 1-cyclopentyl-6-(3,4-trans)-4-methyl-1-[3-(1H-pyrazol-1-yl)benzyl]pyrrolidin-3-yl}- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0143] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(2-methylpyridin-4-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0144] 6-[(3,4-trans)-1-(2-chloro-6-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0145] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0146] 1-cyclopentyl-6-{(3,4-trans)-1-[2-(difluoromethoxy)benzyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-]pyrimidin-4-one

[0147] 1-cyclopentyl-6-{(3,4-trans)-1-[(2-ethoxypyridin-3-yl)methyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0148] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(4,5,6,7-tetrahydropyrazolo[1,5-a]pyridin-3- ylmethyl)pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0149] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dihydro-1,4-benzodioxin-6-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0150] 1-cyclopentyl-6-{(3,4-trans)-1-[4-(1H-imidazol-1-yl)benzyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0151] 1-cyclopentyl-6-[(3,4-trans)-1-(2,5-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0152] 1-cyclopentyl-6-[(3,4-trans)-1-(4-methoxy-3-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0153] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dihydro-1-benzofuran-7-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0154] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0155] 1-cyclopentyl-6-[(3,4-trans)-1-(5-fluoro-2-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0156] 1-cyclopentyl-6-[(3,4-trans)-1-(2-fluoro-4-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0157] 1-cyclopentyl-6-[(3,4-trans)-1-(3-fluoro-4-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0158] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(2-methyl-1,3-thiazol-5- yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0159] 1-cyclopentyl-6-{(3,4-trans)-1-[(4-isopropyl-1,3-thiazol-2-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0160] 1-cyclopentyl-6-{(3,4-trans)-1-[(1,3-dimethyl-1H-pyrazol-5-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0161] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-difluoro-4-methylbenzyl)-4-methylpyrrolidin-3- yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0162] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-{[6-(1H-pyrazol-1-yl)pyridin-2- yl]methyl}pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0163] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(4-methylbenzyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0164] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(2-naphthylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0165] 1-cyclopentyl-6-{(3,4-trans)-1-[(2-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0166] 1-cyclopentyl-6-[(3,4-trans)-1-(2-ethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0167] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[4-(1H-1,2,4-triazol-1-yl)benzyl]pyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0168] 1-cyclopentyl-6-[(3,4-trans)-1-(3-methoxy-4-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0169] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(1-naphthylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0170] 1-cyclopentyl-6-[(3,4-trans)-1-(3-fluoro-4-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0171] 1-cyclopentyl-6-[(3,4-trans)-1-(2,5-dimethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0172] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(5-methylisoxazol-3-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0173] 1-cyclopentyl-6-[(3,4-trans)-1-(2-fluoro-6-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0174] 1-cyclopentyl-6-[(3,4-trans)-1-(2,4-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0175] 1-cyclopentyl-6-[(3,4-trans)-1-(4-fluoro-3-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0176] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dihydro-1,4-benzodioxin-5-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0177] 6-[(3,4-trans)-1-(2-chloro-4-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0178] 1-cyclopentyl-6-[(3,4-trans)-1-(2,4-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0179] 1-cyclopentyl-6-[(3,4-trans)-1-(3,5-dimethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0180] 1-cyclopentyl-6-[(3,4-trans)-1-(3-ethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0181] 6-[(3,4-trans)-1-(4-chloro-2-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0182] 3-{[(3,4-trans)-3-(1-cyclopentyl-4-oxo-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6- yl)-4-methylpyrrolidin-1-yl]methyl}benzonitrile;

[0183] 1-cyclopentyl-6-[(3,4-trans)-1-(2,5-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0184] 2-{[(3,4-trans)-3-(1-cyclopentyl-4-oxo-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6- yl)-4-methylpyrrolidin-1-yl]methyl}benzonitrile;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0185] 6-[(3,4-trans)-1-(3-chloro-4-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0186] 1-cyclopentyl-6-{(3,4-trans)-1-[4-(difluoromethoxy)benzyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0187] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(3-methylbenzyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0188] 1-cyclopentyl-6-[(3,4-trans)-1-(3,4-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0189] 1-cyclopentyl-6-[(3,4-trans)-1-(2,5-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0190] 6-[(3,4-trans)-1-(3-chloro-2-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0191] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0192] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(1,3-thiazol-2-ylmethyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0193] 1-cyclopentyl-6-[(3,4-trans)-1-(3-fluoro-2-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0194] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(2-methylpyrimidin-5-yl)methyl]pyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0195] 1-cyclopentyl-6-{(3,4-trans)-1-[(2-ethylpyrimidin-5-yl)methyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0196] 1-cyclopentyl-6-[(3,4-trans)-1-(4-isopropylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0197] 1-cyclopentyl-6-{(3,4-trans-1-[(1-ethyl-1H-pyrazol-4-yl)methyl]-4-methylpyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0198] 1-cyclopentyl-6-{(3,4-trans)-1-[(4-methoxypyridin-3-yl)methyl]-4-methylpyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0199] 1-cyclopentyl-6-[(3,4-trans)-1-(isoxazol-5-ylmethyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0200] 1-cyclopentyl-6-[(3,4-trans)-1-(4-ethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0201] 1-cyclopentyl-6-[(3,4-trans)-1-{[6-(1-hydroxy-1-methylethyl)pyridin-3-yl]methyl}-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0202] 1-cyclopentyl-6-{(3,4-trans)-1-[(2,2-dimethyl-2,3-dihydro-1-benzofuran-5- yl)methyl]-4-methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0203] 1-cyclopentyl-6-[(3,4-trans)-1-(3,4-dimethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0204] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(5-methylpyrazin-2-yl)methyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0205] 1-cyclopentyl-6-[(3,4-trans)-1-(imidazo[1,2-a]pyridin-2-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0206] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(2-phenyl-1,3-oxazol-4- yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0207] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(2-methylbenzyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0208] 1-cyclopentyl-6-[(3,4-trans)-1-(2-isopropoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0209] 6-[(3,4-trans)-1-(cinnolin-3-ylmethyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0210] 1-cyclopentyl-6-{(3,4-trans)-1-[3-(difluoromethoxy)benzyl]-4-methylpyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0211] 1-cyclopentyl-6-[(3,4-trans)-1-(4-fluoro-3-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0212] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[4-(1H-pyrazol-1-yl)benzyl]pyrrolidin-3-yl}- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0213] 1-cyclopentyl-6-{(3,4-trans)-1-[(2,7-dimethylimidazo[1,2-a]pyridin-3-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0214] 1-cyclopentyl-6-[(3,4-trans)-1-(3,5-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0215] 1-cyclopentyl-6-[(3,4-trans)-1-(4-isopropoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0216] 1-cyclopentyl-6-[(3,4-trans)-1-{[2-(1-hydroxy-1-methylethyl)pyridin-4-yl]methyl}-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0217] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(4,5,6,7-tetrahydro-1,3-benzothiazol-2- ylmethyl)pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0218] 1-cyclopentyl-6-[(3,4-trans)-1-(mesitylmethyl)-4-methylpyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0219] 1-cyclopentyl-6-[(3,4-trans)-1-(2,6-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0220] 4-{[(3,4-trans)-3-(1-cyclopentyl-4-oxo-4,5-dihydro-1H-pyrazolo[3,4-d]pyrimidin-6- yl)-4-methylpyrrolidin-1-yl]methyl}benzonitrile;

[0221] 1-cyclopentyl-6-[(3,4-trans)-1-(2-fluoro-5-methoxybenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0222] 1-cyclopentyl-6-[(3,4-trans)-1-(2,6-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0223] 1-cyclopentyl-6-{(3,4-trans)-1-[(4-methoxy-3,5-dimethylpyridin-2-yl)methyl]-4- methylpyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0224] 1-cyclopentyl-6-[(3,4-trans)-1-(3,5-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0225] 1-cyclopentyl-6-[(3,4-trans)-1-(3,4-dimethylbenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0226] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(1-methyl-1H-benzimidazol-2-yl) methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0227] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(4-methyl-3,4-dihydro-2H-1,4-benzoxazin- 7-yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0228] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(3-phenylpropyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0229] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[2-(trifluoromethyl)benzyl]pyrrolidin-3-yl}- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0230] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(4,4,4-trifluorobutyl)pyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0231] 1-cyclopentyl-6-[(3,4-trans)-1-(3-methoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0232] 1-cyclopentyl-6-[(3,4-trans)-1-(cyclopentylmethyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0233] 1-cyclopentyl-6-[(3,4-trans)-1-(2,4-dimethoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0234] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[4-(morpholin-4-ylmethyl)benzyl]pyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0235] 6-[(3,4-trans)-1-(2,1,3-benzothiadiazol-5-ylmethyl)-4-methylpyrrolidin-3-yl]-1- cyclopentyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0236] 6-{(3,4-trans)-1-[2-(benzyloxy)ethyl]-4-methylpyrrolidin-3-yl}-1-cyclopentyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0237] 1-cyclopentyl-6-[(3,4-trans)-1-(2,6-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0238] 1-cyclopentyl-6-[(3,4-trans)-1-(2-methoxybenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0239] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(3,5,6-trimethylpyrazin-2- yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0240] 1-cyclopentyl-6-[(3,4-trans)-1-(2,4-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0241] 1-cyclopentyl-6-[(3,4-trans)-4-methyl-1-(5,6,7,8-tetrahydro-4H-pyrazolo[1,5- a]azepin-3-ylmethyl)pyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0242] 1-cyclopentyl-6-[(3,4-trans)-1-(3-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0243] 1-cyclopentyl-6-[(3,4-trans)-1-(2,3-dihydro-1-benzofuran-5-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0244] 1-cyclopentyl-6-[(3,4-trans)-1-(2-methoxy-5-methylbenzyl)-4-methylpyrrolidin-3-yl]- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0245] 1-cyclopentyl-6-[(3,4-trans)-1-(2-fluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5-dihydro- 4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0246] 6-[(3,4-trans)-1-(2-chlorobenzyl)-4-methylpyrrolidin-3-yl]-1-cyclopentyl-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0247] 1-cyclopentyl-6-[(3,4-trans)-1-(3,4-dichlorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0248] 6-[(3,4-trans)-1-(2,1,3-benzothiadiazol-4-ylmethyl)-4-methylpyrrolidin-3-yl]-1- cyclopentyl-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0249] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(2-propylpyrimidin-5-yl)methyl]pyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0250] 1-cyclopentyl-6-{(3,4-trans)-1-[(1-ethyl-1H-pyrazol-5-yl)methyl]-4-methylpyrrolidin- 3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0251] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[2-(trifluoromethoxy)benzyl]pyrrolidin-3- yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0252] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[4-(trifluoromethyl)benzyl]pyrrolidin-3-yl}- 1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0253] 1-cyclopentyl-6-{(3,4-trans)-4-methyl-1-[(1-methyl-1H-imidazo[4,5-c]pyridin-2- yl)methyl]pyrrolidin-3-yl}-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one;

[0254] 1-cyclopentyl-6-[(3,4-trans)-1-(3,5-difluorobenzyl)-4-methylpyrrolidin-3-yl]-1,5- dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; or

[0255] 1-cyclopentyl-6-[(3,4-trans)-1-(5,6-dihydro-4H-pyrrolo[1,2-b]pyrazol-3-ylmethyl)-4- methylpyrrolidin-3-yl]-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one; or a pharmaceutically acceptable salt thereof.

[0256] In one preferred embodiment, the compound is 6-((3S,4S)-4-methyl-1-(pyrimidin-2- ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one ("Compound A”) or a pharmaceutically acceptable salt thereof.

[0257] The compounds of the application have been surprisingly found to show pharmacological activity including selectively inhibiting PDE9 that makes them suitable for the treatment, prevention and / or control of in treating conditions that may be regulated or normalized by inhibition of PDE9.

[0258] The compounds and intermediates of the present application may be named according to either the IUPAC (International Union for Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature systems.

[0259] The compounds of Formula (I) may contain asymmetric or chiral centers and, therefore, exist in different stereoisomeric forms. Those skilled in the art will appreciate that, unless otherwise specified, all stereoisomers (e.g., enantiomers and diastereoisomers, and racemic mixtures thereof) of the novel compounds and intermediates described, illustrated and / or discussed herein are within the scope of the claimed application. In addition, unless otherwise specified, the present application embraces all geometric and positional isomers. The (3S,4S) enantiomer of the core pyrrolidinyl configuration is preferred.

[0260] Diasteriomeric mixtures can be separated into their individual diastereomers on the basis of their physical chemical differences by methods well-known to those of ordinary skill in the art, such as by chromatography and / or fractional crystallization. Enantiomers can be separated by converting the enantiomeric mixture into a diasteriomeric mixture by reaction with an appropriate optically active compound (e.g., alcohol), separating the diastereomers and converting (e.g., hydrolyzing) the individual diastereomers to the corresponding pure enantiomers. Additional methods include resolution of racemic mixtures using chiral salts, as well as chiral chromatography.

[0261] Those skilled in the art will further recognize that the compounds of Formula (I) can exist in crystalline form as hydrates wherein molecules of water are incorporated within theAttorney Docket No.: CDPH-003 / 001WO 341287-2404 crystal structure thereof and as solvates wherein molecules of a solvent are incorporated therein. All such hydrate and solvate forms are considered part of this application.

[0262] Practitioners will appreciate that certain compounds of Formula (I) may exist as tautomeric isomers, i.e., that equilibrium exists between two isomers which are in rapid equilibrium with each other. A common example of tautomerism is keto-enol tautomerism, i.e.,

[0263] The degree to which one tautomer is present over the other depends upon various factors, including substitution pattern and solvent type. Other examples in accordance with the present application will be recognized by those skilled in the art. All tautomeric forms of Formula (I) are included within the scope of the application unless otherwise specified.

[0264] The present application also includes prodrugs of the compounds of the application. The term “prodrug” refers to a drug precursor which, following administration, releases the drug in vivo via a chemical or physiological process (e.g., upon being brought to physiological pH or through enzymatic activity). The prodrug may itself be biologically active, or may be converted to a biologically active compound (e.g. by metabolism or hydrolysis) during its residence time in the body. A discussion of the preparation and use of prodrugs is provided by Higuchi & Stella, “Prodrugs as Novel Delivery Systems”, Vol. 14 of the A.C.S. Symposium Series, and in “Bioreversible Carriers in Drug Design,” ed. Edward B. Roche, American Pharmaceutical Association and Pergamon Press, 1987. All prodrugs of the various compounds of Formula (I) are within the scope of the present application.

[0265] The present application also embraces isotopically-labeled compounds of Formula (I) that are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of Formula (I) include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, such as2H,3H,13C,14C,15N,18O,17O,31P,32P,35S,18F and36Cl, respectively. The compounds of Formula (I), and pharmaceutically acceptable salts thereof, that contain the aforementioned isotopes and / or other isotopes of the other atoms are within the scope of the instant application.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0266] Certain isotopically-labeled compounds of Formula (I), for example those into which radioactive isotopes such as3H and14C are incorporated, are useful in drug and / or substrate tissue distribution assays. Tritiated, i.e.,3H and14C, isotopes are particularly preferred for their ease of preparation and detectability. Furthermore, substitution with heavier isotopes such as deuterium, i.e.,2H, may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life, or reduced dosage requirements and, hence, may be preferred in some circumstances. Isotopically-labeled compounds of Formula (I), and pharmaceutically acceptable salts thereof, can be generally prepared by carrying out analogous procedures to those disclosed in the Schemes and / or in the Examples below, by substituting a readily available isotopically-labeled reagent for a non-isotopically labeled reagent. Definitions

[0267] Certain terms used herein are generally defined as follows:

[0268] The carbon atom content of the various hydrocarbon-containing moieties herein may be indicated by a prefix designating the minimum and maximum number of carbon atoms in the moiety. Thus, for example, (C1-C6)alkyl refers to an alkyl group of one to six carbon atoms inclusive.

[0269] The term “alkoxy” refers to a straight or branched, monovalent, saturated aliphatic hydrocarbon radical bonded to an oxygen atom that is attached to a core structure. Examples of alkoxy groups include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert- butoxy, pentoxy, and the like.

[0270] The term “alkyl” means a saturated monovalent straight or branched aliphatic hydrocarbon radical. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, sec-butyl, pentyl, isopentyl, neopentyl, hexyl, isohexyl, and the like.

[0271] The term “alkenyl” means a partially unsaturated straight or branched aliphatic hydrocarbon radical having one or more double bonds. Examples of alkenyl groups include ethenyl (also known as “vinyl”), allyl, 1-propenyl, isopropenyl, n-butenyl, n-pentenyl, and the like. The term “alkenyl” embraces radicals having “cis” and “trans” orientations, or alternatively, “Z” and “E” orientations.

[0272] The term “alkynyl” means a partially unsaturated straight or branched aliphatic hydrocarbon radical having one or more double bonds. Examples of alkynyl groups include 1-propynyl, 2-propynyl (also known as “propargyl”), 1-butynyl, 2-butynyl, 1-pentynyl, and the like.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0273] The term “aryl” denotes a monocyclic or polycyclic aromatic ring system, for example, anthracenyl, benzyl, fluorenyl, indenyl, naphthyl, phenanthrenyl, phenyl and the like. The term “aryl” is also intended to include the partially hydrogenated derivatives of such ring systems, e.g. 1,2,3,4-tetrahydronaphthyl.

[0274] The term “aryloxy” denotes an aryl radical bonded to an oxygen atom that is attached to a core structure, such as benzyloxy.

[0275] The term “cycloalkyl” denotes a saturated monocyclic or bicyclic cycloalkyl group. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like.

[0276] The term “halogen” or “halo” represents chlorine, bromine, fluorine and iodine atoms and radicals.

[0277] The term “haloalkyl” refers to an alkyl or cycloalkyl substituent wherein at least one hydrogen radical is replaced with a halogen radical. Where more than one hydrogen is replaced with halogen, the halogens may be the same or different. Examples of haloalkyl radicals include trifluoromethyl, 2,2,2-trifluoroethyl, 4,4,4-trifluorobutyl, 4,4- difluorocyclohexyl, chloromethyl, dichloromethyl, trichloromethyl, 1-bromoethyl, and the like.

[0278] The term “haloalkoxy” refers to an alkoxy radical in which at least one hydrogen radical is replaced with a halogen radical. Where more than one hydrogen is replaced with halogen, the halogens may be the same or different. Examples of haloalkoxy radicals include difluoromethoxy, trifluoromethoxy, 2,2,2-trifluoroethoxy, chloromethoxy, bromomethoxy, and the like.

[0279] The term “heteroaryl” as used herein includes heterocyclic unsaturated ring systems containing one or more heteroatoms such as nitrogen, oxygen, and sulfur. If the heteroaryl group contains more than one heteroatom, the heteroatoms may be the same or different. The heteroaryl radicals may be bonded via a carbon atom or a heteroatom. The term “heteroaryl” is also intended to include the partially hydrogenated derivatives of such ring systems. Examples of heteroaryl groups include furanyl (also known as “furyl”), imidazolinyl, imidazolyl (also known as “1,3-diazolyl”), indolyl, oxadiazolyl, oxazinyl, oxazolyl, isoxazolyl, pyranyl, pyrazinyl (also known as “1,4-diazinyl”), pyrazolyl (also known as “1,2- diazolyl”), pyrazolinyl, pyrazyl, pyridazinyl (also known as “1,2-diazinyl”), pyridyl (also known as pyridinyl), pyrimidinyl (also known as “1,3 diazinyl” and “pyrimidyl”), pyrrolyl, thiadiazinyl, thiadiazolyl, thiatriazolyl, thiazolyl, isothiazolyl, thienyl, thiofuranyl (also known as “thiophenyl”), thiopyranyl, triazinyl, triazolyl, and the like.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0280] The term “heteroaryl” also embraces radicals in which 2 or 3 rings are fused together, wherein at least on such ring contains a heteroatom as a ring atom, including radicals wherein (a) a heterocycloalkyl ring is fused with an aryl or heteroaryl ring, or (b) a cycloalkyl ring is fused with a heteroaryl ring. Examples of 2-fused ring heteroaryls include benzodioxinyl, dihydrobenzodioxinyl, benzofuranyl, dihydrobenzofuranyl, isobenzofuranyl, benzimidazolyl, benzothiadiazolyl, tetrahydrobenzothiadiazolyl, benzothiazolyl, benzothienyl (also known as “benzothiophenyl,” “thionaphthenyl,” and “benzothiofuranyl”), benzoxazinyl, dihydrobenzoxazinyl, benzoxazolyl, chromanyl, isochromanyl, chromenyl, cinnolinyl (also known as “1,2-benzodiazinyl”), imidazopyridinyl (e.g. imidazo[1,2-a]pyridinyl or imidazo[4,5-c]pyridinyl), indazolyl, indolinyl, isoindolinyl, indolizinyl, indolyl, isoindolyl, naphthyridinyl, oxathiolopyrrolyl, pteridinyl, pthalazinyl, purinyl (also known as “imidazo[4,5-d]pyrimidinyl”), pyranopyrrolyl, pyrazoloazepinyl, tetrahydropyrazoloazepinyl (e.g. tetrahydropyrazolo[1,5-a]azepinyl), pyrazolopyridinyl, tetrahydropyrazolopyridinyl (e.g. tetrahydropyrazolo[1,5-a]pyridinyl), pyrazolopyrimidinyl (e.g. pyrazolo[3,4-d]pyrimidinyl), pyridopyrazinyl (e.g. pyrido[2,3-b]pyrazinyl), pyridopyridinyl, pyrrolopyrazolyl, dihydropyrrolopyrazolyl (e.g. dihydropyrrolo[1,2-b]pyrazolyl), quinazolinyl (also known as “1,3-benzodiazinyl”), quinolinyl (also known as “1-benzazinyl”), isoquinolinyl (also known as “2-benzazinyl”), quinolizinyl, quinolyl, isoquinolyl, quinoxalinyl, dithianaphthalenyl, thienofuranyl (e.g. thieno[3,2-b]furanyl), and the like.

[0281] Examples of 3-fused ring heteroaryls include acridinyl, diazaanthryl, triazaphenanthrene, carbazolyl, carbolinyl, furocinnolinyl, perimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, thianthrenyl, xanthenyl, and the like.

[0282] The term “heterocycloalkyl” denotes a saturated monocyclic or polycyclic cycloalkyl group, in which at least one of the carbon atoms is replaced with a heteroatom such as nitrogen, oxygen or sulfur. If the heterocycle contains more than one heteroatom, the heteroatoms may be the same or different. The heterocycloalkyl radicals may be bonded via a carbon atom or a heteroatom. Examples of heterocycloalkyl groups include azetidinyl, dioxacyclohexyl, 1,3-dioxolanyl, imidazolidinyl, morpholinyl, piperazinyl, piperidinyl, pyrazolidinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, thiazanyl, and the like.

[0283] A cyclic group may be bonded to another group in more than one way. If no particular bonding arrangement is specified, then all possible arrangements are intended. For example, the term “pyridyl” includes 2-, 3- or 4-pyridyl (2-, 3-, or 4-pyridinyl).Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0284] The term “oxo” means a carbonyl group formed by the combination of a carbon atom and an oxygen atom.

[0285] The term “salts” refers to both organic and inorganic salts of a compound of Formula (I). Such salts can be prepared in situ during the final isolation and purification of a compound, or by separately reacting a compound, prodrug or stereoisomer of Formula (I) with a suitable organic or inorganic acid or base and isolating the salt thus formed. Representative anionic salts include hydrobromide, hydrochloride, hydroiodide, sulfate, bisulfate, nitrate, acetate, trifluoroacetate, oxalate, besylate, palmitate, pamoate, malonate, stearate, laurate, malate, borate, benzoate, lactate, phosphate, hexafluorophosphate, benzene sulfonate, tosylate, formate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate and laurylsulphonate salts and the like. Representative cationic salts include sodium, potassium, calcium, and magnesium salts and the like. See generally, e.g., Berge, et al., J. Pharm. Sci., 66, 1-19 (1977).

[0286] A salt of a compound of Formula (I) may be readily prepared by mixing together solutions of a compound of Formula (I) and the desired acid or base, as appropriate. The salt may precipitate from solution and be collected by filtration or may be recovered by evaporation of the solvent.

[0287] The term “radical” denotes a group of atoms that behaves as a single reactant in a chemical reaction, e. g., an organic radical is a group of atoms that imparts characteristic properties to a compound containing it, or which remains unchanged during a series of reactions or transformations.

[0288] The symbol “—” represents a covalent bond.

[0289] The phrase “reaction-inert solvent” or “inert solvent” refers to a solvent, or mixture of solvents, that does not interact with starting materials, reagents, intermediates or products in a manner that adversely affects their desired properties.

[0290] The terms “treat,” “treating,” “treated” or “treatment” as used herein includes preventative (e.g., prophylactic), palliative or curative uses or results. Pharmaceutical Compositions

[0291] The application also includes pharmaceutical compositions comprising an amount of a compound of Formula (I), or a pharmaceutically acceptable salt of the compound, and optionally a pharmaceutically acceptable vehicle, carrier or diluent. In some embodiments, the pharmaceutical composition consists essentially of a compound of Formula (I) or a pharmaceutically acceptable salt thereof. In a preferred embodiment, the pharmaceuticalAttorney Docket No.: CDPH-003 / 001WO 341287-2404 composition is of an amount effective at inhibiting the enzyme PDE9 in a mammal. In another preferred embodiment, the mammal is a human.

[0292] Methods of preparing various pharmaceutical compositions with amounts of active ingredients are known, or will be apparent in light of this disclosure, to those skilled in this art. See, for example, Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., 19th Ed. (1995).

[0293] Suitable pharmaceutical carriers, vehicles and diluents for such compositions include inert solid diluents or fillers, sterile aqueous solutions and various organic solvents. The pharmaceutical compositions formed by combining a compound of this application and pharmaceutically acceptable carriers, vehicles or diluents are readily administered in a variety of dosage forms such as tablets, powders, lozenges, syrups, injectable solutions and the like.

[0294] Solid dosage forms for oral administration include capsules, tablets, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert conventional pharmaceutical excipient (or carrier) such as sodium citrate, calcium carbonate, or dicalcium phosphate, or (a) fillers or extenders, such as for example, starches, lactose, sucrose, mannitol and silicic acid; (b) binders, such as for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, such as for example, glycerol; (d) disintegrating agents, such as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, such as for example, paraffin; (f) absorption accelerators, such as for example, quaternary ammonium compounds; (g) wetting agents, such as for example, cetyl alcohol and glycerol monostearate; (h) adsorbents, such as for example, kaolin and bentonite; and / or (i) lubricants, such as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or mixtures thereof. In the case of capsules and tablets, the dosage forms may further comprise buffering agents.

[0295] Solid dosage forms may be formulated as modified release and pulsatile release dosage forms containing excipients such as those detailed above for immediate release dosage forms together with additional excipients that act as release rate modifiers, these being coated on and / or included in the body of the device. Release rate modifiers include, but are not limited to, hydroxypropylmethyl cellulose, methyl cellulose, sodium carboxymethylcellulose, ethyl cellulose, cellulose acetate, polyethylene oxide, xanthan gum, ammoniomethacrylate copolymer, hydrogenated castor oil, carnauba wax, paraffin wax, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, methacrylic acidAttorney Docket No.: CDPH-003 / 001WO 341287-2404 copolymer and mixtures thereof. Modified release and pulsatile release dosage forms may contain one or a combination of release rate modifying excipients.

[0296] The pharmaceutical compositions of the application may further comprise fast dispersing or dissolving dosage formulations (FDDFs). The terms dispersing or dissolving as used herein to describe FDDFs are dependent upon the solubility of the drug substance used i.e., where the drug substance is insoluble, a fast dispersing dosage form may be prepared, and where the drug substance is soluble, a fast dissolving dosage form may be prepared.

[0297] Solid compositions of a similar type may also be employed as fillers in soft or hard filled gelatin capsules using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

[0298] Solid dosage forms such as tablets, dragees, capsules, and granules can be prepared with coatings and shells, such as enteric coatings and others well-known to one of ordinary skill in the art. They may also comprise opacifying agents, and can also be of such composition that they release the active compound(s) in a delayed, sustained or controlled manner. Examples of embedding compositions that can be employed are polymeric substances and waxes. The active compound(s) can also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

[0299] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compounds, the liquid dosage form may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethanol, isopropanol, ethyl carbonate, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil, and sesame seed oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances, and the like.

[0300] In addition to the active compound(s), the pharmaceutical composition may further include suspending agents, such as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar, and tragacanth, or mixtures of these substances, and the like. Sweeteners, flavoring, and perfuming agents may also be included.

[0301] The pharmaceutical compositions of the application may further comprise adjuvants, such as preserving, wetting, emulsifying and dispersing agents. Prevention of microorganism contamination of the instant compositions can be accomplished with various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid and the like. ItAttorney Docket No.: CDPH-003 / 001WO 341287-2404 may also be desirable to include isotonic agents, for example, sugars, sodium chloride and the like. Prolonged absorption of injectable pharmaceutical compositions may be affected by the use of agents capable of delaying absorption, for example, aluminum monostearate and gelatin.

[0302] For parenteral administration, solutions in sesame or peanut oil, aqueous propylene glycol, or in sterile aqueous solutions may be employed. Such aqueous solutions should be suitably buffered if necessary and the liquid diluent first rendered isotonic with sufficient saline or glucose. These aqueous solutions are especially suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration. In this connection, the sterile aqueous media employed are all readily available by standard techniques known to those skilled in the art.

[0303] For intranasal administration or administration by inhalation, the compounds of Formula (I) are conveniently delivered in the form of a solution or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve to deliver a metered amount. The pressurized container or nebulizer may contain a solution or suspension of a compound of this application. Capsules and cartridges (made, for example, from gelatin) for use in an inhaler or insulator may be formulated containing a powder mix of a compound or compounds of the application and a suitable powder base such as lactose or starch.

[0304] Pharmaceutical compositions of the present application may also be configured for treatments in veterinary use, where a compound of the present application, or a veterinarily acceptable salt thereof, or veterinarily acceptable solvate or pro-drug thereof, is administered as a suitably acceptable formulation in accordance with normal veterinary practice and the veterinary practitioner will determine the dosing regimen and route of administration which will be most appropriate for a particular animal.

[0305] In general, the compounds of Formula (I), and pharmaceutically acceptable salts thereof, may be prepared according to the exemplary routes disclosed in the Schemes and Examples disclosed in U.S. Patent No. 7,964,607, which is incorporated herein by reference in its entirety, as well as by other conventional preparative procedures known, or apparent in light of the instant disclosure, to one of ordinary skill in the art.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Methods of Treatment

[0306] The application also includes methods of treating chronic heart failure in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of: (a) a compound of Formula (I), or a pharmaceutically acceptable salt thereof; or (b) a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in a pharmaceutically acceptable vehicle, carrier or diluent; either alone or in combination with a second agent described above.

[0307] The application also includes use of (a) a compound of Formula (I), or a pharmaceutically acceptable salt thereof; or (b) a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for treating chronic heart failure in a subject in need thereof.

[0308] The application also includes (a) a compound of Formula (I), or a pharmaceutically acceptable salt thereof; or (b) a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceutically acceptable salt thereof, for use in treating chronic heart failure in a subject in need thereof.

[0309] In some embodiments, the compound of Formula (I), or a pharmaceutically acceptable salt thereof is 6-((3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl)-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one (“Compound A”) or a pharmaceutically acceptable salt thereof. Dosage and Administration

[0310] The appropriate dosage regimen, the amount of each dose administered and the intervals between doses of the compound will depend, among others, upon the compound of Formula (I) of this application being used, the type of pharmaceutical compositions being used, the characteristics of the subject being treated and the type and severity of the conditions to be treated. In general, an effective dose for compounds of Formula (I) or pharmaceutically acceptable salts thereof, is in the range of from about 5 mg to about 100 mg per day. In some embodiments, the compound of Formula (I) is administered at a dose of 0.1 mg to 0.5 mg, 0.5 mg to 1 mg, 1 mg to 5 mg, 5 mg to 90 mg, 5 mg to 80 mg, 5 mg to 70 mg, 5 mg to 60 mg, 5 mg to 50 mg, 5 mg to 45 mg, 5 mg to 40 mg, 5 mg to 35 mg, 5 mg to 30 mg, 5 mg to 25 mg, 5 g to 20 mg, 5 mg to 15 mg, 5 mg to 10 mg, 10 mg to 50 mg, 50 g to 100 mg, 100 mg to 150 mg, 150 mg to 200 mg, 200 mg to 250 mg, or 250 mg to 300 mg. In some embodiments, the compound of Formula (I) is administered at a dose of 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, or 300 mg.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0311] Administration may be in single (e.g., once daily) or multiple doses. In some embodiments, the compound of Formula (I) is administered once every 4 hours, once every 8 hours, once every 12 hours, once every 18 hours, or once every 24 hours, once daily, once every other day, one every three days, or once a week. In some embodiments, 25 mg of the compound of Formula (I) are administered every 12 hours. In some embodiments, 50 mg of the compound of Formula (I) are administered per day. In preferred embodiments, the compound of Formula (I) is administered twice daily. In other preferred embodiments, the compound of Formula (I) is administered every 12 hours. In some embodiments, the administration is continued for 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 6 months, 8 months, 10 months, one year, or more. The administration may be continued until the patient improves. In some embodiments, the compound of Formula (I) is administered for the rest of the patient’s life. The administration can be continuous or intermittent.

[0312] Some variability in the general dosage range may be required depending upon the age and mass of the subject being treated, the intended route of administration, the particular compound being administered, and the like. The determination of dosage ranges and optimal dosages for a particular mammalian subject is within the ability of a skilled person having benefit of the instant disclosure.

[0313] The compounds of Formula (I) may be administered by a variety of conventional routes of administration, including oral, buccal, sublingual, ocular, topical (e.g., transdermal), parenteral (e.g., intravenous, intramuscular, or subcutaneous), rectal, intracisternal, intravaginal, intraperitoneal, intravesical, local (e.g., powder, ointment, or drop), nasal and / or inhalation dosage forms or using a “flash” formulation, i.e., allowing the medication to dissolve in the mouth without the need to use water. As will be recognized by one of skill in the art, the appropriate dosage regimen, the amount of each dose administered and the intervals between doses of the compound will depend upon the compound of Formula (I), or the prodrug thereof, being used, the type of pharmaceutical compositions being used, the characteristics of the subject being treated, and / or the severity of the conditions being treated. In preferred embodiments, the compound of Formula (I) is administered orally.

[0314] The application also includes methods of inhibiting PDE9 in a mammal comprising administering to the mammal in need of such inhibition a PDE9 inhibiting amount of: (a) a compound of Formula (I), or a pharmaceutically acceptable salt thereof; or (b) a pharmaceutical composition comprising a compound of Formula (I), or a pharmaceuticallyAttorney Docket No.: CDPH-003 / 001WO 341287-2404 acceptable salt thereof, in a pharmaceutically acceptable vehicle, carrier or diluent; either alone or in combination with a second agent as described above. Patient Population

[0315] The methods of treatment described herein are useful for the treatment of heart conditions, in particular those associated with abnormal PDE9 / cGMP signaling. In preferred embodiments, a patient treated in accordance with a method described herein is afflicted with heart failure.

[0316] In some embodiments, a subject treated in accordance with the methods described herein has chronic heart failure. In some embodiments, the patient has chronic heart failure with reduced ejection fraction (EF). In some embodiments, the patient has chronic heart failure with preserved EF. In some embodiments, the patient has chronic heart failure with midrange EF (between 40% and 60% or between 40% and 50%). In some embodiments, the patient has chronic heart failure with super EF (above 60%). In some embodiments, the patient has left ventricular hypertrophy. In some embodiments, the patient has congestive heart failure.

[0317] In some embodiments, the subject has an ejection fraction (EF) of less than 40% as determined by echocardiography. In some embodiments, the subject has an EF of 40% to 60% as determined by echocardiography. In some embodiments, the subject has an EF of more than 60% as determined by echocardiography. In some embodiments, the subject has an EF of less than 60% as determined by echocardiography. In some embodiments, the subject has an EF of more than 40% and left atrial enlargement by echocardiography. The treatment of the chronic heart failure may result in the improvement in the EF.

[0318] In some embodiments, the subject has plasma NT-proBNP levels of at least 600 pg / ml. In some embodiments, the subject has plasma NT-proBNP levels of at least 300 pg / ml. In some embodiments, the subject as atrial fibrillation or flutter and an NT-proBNP level of ≥1000 pg / mL. In some embodiments, the subject as atrial fibrillation or flutter and an NT-proBNP level of ≥500 pg / mL. The treatment of the chronic heart failure may result in the improvement of NT-proBNP levels in the patient (e.g., plasma NT-proBNP, or serum NT- proBNP levels) .

[0319] The terms “subject” and “patient” are used interchangeably herein. In some embodiments, the patient is human. Determining Efficacy

[0320] The term “therapeutically effective amount” as it refers to the compound of Formula (I) here describes an amount of the compound of Formula (I) that is sufficient to bring aboutAttorney Docket No.: CDPH-003 / 001WO 341287-2404 one or more desired treatment effects. Such effects include, for example, an improvement of a symptom or of a clinical marker. Any suitable method known in the art or described herein may be used to measure the efficacy of a method of treatment described herein.

[0321] In some embodiments, a method described here results in a reduction in N-terminal pro b-type natriuretic peptide (NT-proBNP) levels and / or in b-type natriuretic peptide (BNP) in the subject’s plasma. NT-proBNP and BNP are circulating biomarkers that can be detected in the blood. Both originate from the same pro-peptide produced by the heart in response to myocyte stretching. BNP induces diuresis, vasodilatation, and decreased renin and aldosterone secretion (see Hall, J Card Fail 2005 Jun;11(5 Suppl):S81-3). Increased levels of NT-proBNP and / or BNP are poor prognostic biomarkers in several types of cardiac disease (Salah et al., Heart 2019; 105:1182-1189). The normal range of NT-proBNP is 0-125 pg / mL for patients under 74 years of age, and 0-450 pg / mL for patients about 74 years of age. Levels of 450 mg / mL in patients under 50 and levels above 900 pg / mL in patients of 50 are older indicate compromised heart function. BNP levels of less than 100 pg / mL are considered normal, and levels above 100 pg / mL indicate compromised heart function.

[0322] In some embodiments, a method of treatment described herein results in a decrease of NT-proBNP levels of about 10- 20%, about 20%-30%, about 30%-40%, or about 40%-50%, as measured in the plasma of the subject. In some embodiments, a method of treatment described herein results in a decrease of NT-proBNP levels of about 10%, about 20%, about 30%, about 40%, about 50%, or more than 50% as measured in the plasma of the subject.

[0323] In some embodiments, a method of treatment described herein results in a decrease of BNP levels of about 10- 20%, about 20%-30%, about 30%-40%, or about 40%-50%, as measured in the plasma of the subject. In some embodiments, a method of treatment described herein results in a decrease of BNP levels of about 10%, about 20%, about 30%, about 40%, about 50%, or more than 50% as measured in the plasma of the subject.

[0324] In some embodiments, a method of treatment described herein results in an increase in cGMP levels in the plasma or urine of the subject. 3’, 5’-cyclic guanosine monophosphate (cGMP) is a second messenger involved in various signaling pathways which produced by guanylate cyclases. cGMP is broken down by phosphodiesterases 5 and 9 in the myocardium (PDE5 and PDE9). Treatment with the compound of Formula (I), which inhibits PDE9, is believed to increase cGMP levels.

[0325] In some embodiments, a method of treatment described herein results in an increase of cGMP levels of about 10- 20%, about 20%-30%, about 30%-40%, or about 40%-50%,. In some embodiments, a method of treatment described herein results in an increase of cGMPAttorney Docket No.: CDPH-003 / 001WO 341287-2404 levels of about 10%, about 20%, about 30%, about 40%, about 50%, or more than 50%. In some embodiments, the cGMP levels are measured in the urine of the subject. In some embodiments, the cGMP levels are measured in the blood of the subject. In some embodiments, the cGMP levels are measured in the plasma of the subject.

[0326] In some embodiments, a method of treatment described herein modulate the ratio of cGMP to NT-proBNP and / or the ratio of cGMP to BNP. A decreased cGMP to BNP ratio has been described as a poor prognostic factor in heart failure patients (see Lourenço et al., Eur J Heart Fail. 2009 Feb. 11(2): 185-190).

[0327] In some embodiments, a method of treatment described herein results in an increase of the cGMP to NT-proBNP ratio of about 10-20%, about 20%-30%, about 30%-40%, or about 40%-50%. In some embodiments, a method of treatment described herein results an increase of the cGMP to NT-proBNP ratio of about 10%, about 20%, about 30%, about 40%, about 50%, or more than 50%. In some embodiments, the cGMP levels are measured in the blood of the subject. In some embodiments, the cGMP levels are measured in the plasma of the subject. In some embodiments, the cGMP levels are measured in the urine of the subject. In some embodiments, the NT-proBNP levels are measured in the plasma of the subject.

[0328] In some embodiments, a method of treatment described herein results in an increase of the cGMP to BNP ratio of about 10-20%, about 20%-30%, about 30%-40%, or about 40%- 50%. In some embodiments, a method of treatment described herein results an increase of the cGMP to BNP ratio of about 10%, about 20%, about 30%, about 40%, about 50%, or more than 50%. In some embodiments, the cGMP levels are measured in the blood of the subject. In some embodiments, the cGMP levels are measured in the plasma of the subject. In some embodiments, the BNP levels are measured in the blood of the subject. In some embodiments, the BNP levels are measured in the plasma of the subject. In some embodiments, the cGMP levels are measured in the urine of the subject.

[0329] In some embodiments, a method of treatment described herein results in an increase in the Kansas City Cardiomyopathy Questionnaire (KCCQ) score of a patient. The KCCQ is an FDA-approved clinical outcome assessment and recommended as a performance measure for quantifying the quality of care in heart failure (see Spertus et al., JACC 2020, 76(20). It can measure the impact of heart failure on patients’ lives and is strongly associated with clinical events over time. Several versions of the KCCQ are available, including a 12-item version (KCCQ-12) and a 23-item version (KCCQ-23). The KCCQ scores range from 1-100, represent health status as follows: 0 to 24: very poor to poor; 25 to 49: poor to fair; 50 to 74: fair to good; and 75 to 100: good to excellent.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0330] In some embodiments, a method of treatment described herein results in an improvement of 5-10 points, 10-15 points, 15-20 points, 20-25 points, 25-30 points, 30-35 points, 35-40 points, 40-45 points, 45-50 points, or more than 50 points on the KCCQ. In some embodiments, a method of treatment described herein results in an improvement of at least 5 points, at least 10 points, at least 15 points, at least 20 points, at least 25 points, at least 30 points, at least 35 points, at least 40 points, at least 45 points, or at least 50 points on the KCCQ.

[0331] The efficacy of a method of treatment described herein may be measured at any suitable time point after administration. In some embodiments, the efficacy is determined 12 weeks after the first administration. In some embodiments, the efficacy is determined 6 months after the first administration. In some embodiments, the efficacy is determined 12 months after the first administration.

[0332] In some embodiments, a method of treatment described herein results in an improvement of mortality in the subject. In some embodiments, a method described herein results in a decrease in mortality of about 10-20%, about 20%-30%, about 30%-40%, or about 40%-50% compared to a subject of similar age and diagnosed with chronic heart failure who is not being treated in accordance with a method described herein. In some embodiments, the mortality is heart failure mortality. In some embodiments, the mortality is cardiovascular mortality. In some embodiments, the mortality is all-cause mortality.

[0333] In some embodiments, a method of treatment described herein results in a reduction in heart failure hospitalization in the patient. In some embodiments, the method reduces heart failure hospitalization by 10-20%, about 20%-30%, about 30%-40%, or about 40%-50% compared to a subject of similar age and diagnosed with chronic heart failure who is not being treated in accordance with a method described herein. Combination Therapies

[0334] The present application includes the use of a combination of a PDE9 inhibitor compound as provided in Formula (I) and one or more additional pharmaceutically active agent(s). If a combination of active agents is administered, then they may be administered sequentially or simultaneously, in separate dosage forms or combined in a single dosage form. Accordingly, the present application also includes pharmaceutical compositions comprising an amount of: (a) a first agent comprising a compound of Formula (I) or a pharmaceutically acceptable salt of the compound; (b) a second pharmaceutically active agent; and (c) a pharmaceutically acceptable carrier, vehicle or diluent.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0335] Various pharmaceutically active agents may be selected for use in conjunction with the compounds of Formula (I), depending on the disease, disorder, or condition to be treated. Pharmaceutically active agents that may be used in combination with the compositions of the present application include, without limitation:

[0336] (vi) beta-adrenergic receptor blocking agents (beta blockers), such as carteolol, esmolol (BREVIBLOC), labetalol (NORMODYNE, TRANDATE), oxprenolol (LARACOR, TRASACOR), pindolol (VISKEN), propanolol (INDERAL), sotalol (BETAPACE, SOTALEX, SOTACOR), timolol (BLOCADREN, TIMOPTIC), acebutolol (SECTRAL, PRENT), nadolol (CORGARD), metoprolol tartrate (LOPRESSOR), metoprolol succinate (TOPROL-XL), atenolol (TENORMIN), butoxamine, and SR 59230A (Sanofi);

[0337] (x) calcium channel blockers such as nilvadipine (ESCOR, NIVADIL), amlodipine (NORVASC, ISTIN, AMLODIN), felodipine (PLENDIL), nicardipine (CARDENE), nifedipine (ADALAT, PROCARDIA), MEM 1003 and its parent compound nimodipine (NIMOTOP), nisoldipine (SULAR), nitrendipine, lacidipine (LACIPIL, MOTENS), lercanidipine (ZANIDIP), diltiazem (CARDIZEM), verapamil (CALAN, VERELAN), and enecadin (also known as NS-7);

[0338] (xi) catechol O-methyltransferase (COMT) inhibitors, such as tolcapone (TASMAR), entacapone (COMTAN), and tropolone;

[0339] (xii) central nervous system stimulants, such as caffeine, phenmetrazine, phendimetrazine, pemoline, fencamfamine (GLUCOENERGAN, REACTIVAN), fenethylline (CAPTAGON), pipradol (MERETRAN), deanol (also known as dimethylaminoethanol), methylphenidate (DAYTRANA), methylphenidate hydrochloride (RITALIN), dexmethylphenidate (FOCALIN), amphetamine (alone or in combination with other CNS stimulants, e.g. ADDERALL (amphetamine aspartate, amphetamine sulfate, dextroamphetamine saccharate, and dextroamphetamine sulfate)), dextroamphetamine sulfate (DEXEDRINE, DEXTROSTAT), methamphetamine (DESOXYN), lisdexamfetamine (VYVANSE), and benzphetamine (DIDREX);

[0340] (xiii) corticosteroids, such as prednisone (STERAPRED, DELTASONE), prednisolone (PRELONE), predisolone acetate (OMNIPRED, PRED MILD, PRED FORTE), prednisolone sodium phosphate (ORAPRED ODT), methylprednisolone (MEDROL); methylprednisolone acetate (DEPO-MEDROL), and methylprednisolone sodium succinate (A-METHAPRED, SOLU-MEDROL);

[0341] (xiv) dopamine receptor agonists, such as apomorphine (APOKYN), bromocriptine (PARLODEL), cabergoline (DOSTINEX), dihydrexidine, dihydroergocryptine, fenoldopamAttorney Docket No.: CDPH-003 / 001WO 341287-2404 (CORLOPAM), lisuride (DOPERGIN), pergolide (PERMAX), piribedil (TRIVASTAL, TRASTAL), pramipexole (MIRAPEX), quinpirole, ropinirole (REQUIP), and rotigotine (NEUPRO);

[0342] (xv) dopamine receptor antagonists, such as tetrabenazine (NITOMAN, XENAZINE), 7-hydroxyamoxapine, droperidol (INAPSINE, DRIDOL, DROPLETAN), domperidone (MOTILIUM), L-741742, L-745870, raclopride, SCH-23390, ecopipam, SKF-83566, and metoclopramide (REGLAN);

[0343] (xvi) dopamine reuptake inhibitors such as nomifensine maleate (MERITAL), vanoxerine (also known as GBR-12909) and its decanoate ester DBL-583, and amineptine;

[0344] (xvii) gamma-amino-butyric acid (GABA) receptor agonists, such as baclofen (LIORESAL, KEMSTRO), pentobarbital (NEMBUTAL), progabide (GABRENE), and clomethiazole;

[0345] (xviii) immunomodulators such as glatiramer acetate (also known as copolymer-1; COPAXONE), MBP-8298 (synthetic myelin basic protein peptide), dimethyl fumarate, fingolimod (also known as FTY720), roquinimex (LINOMIDE), laquinimod (also known as ABR-215062 and SAIK-MS), ABT-874 (human anti-IL-12 antibody), rituximab (RITUXAN), alemtuzumab (CAMPATH), daclizumab (ZENAPAX), and natalizumab TSABRI);

[0346] (xix) immunosuppressants such as methotrexate (TREXALL, RHEUMATREX), mitoxantrone (NOVANTRONE), mycophenolate mofetil (CELLCEPT), mycophenolate sodium (MYFORTIC), azathioprine (AZASAN, IMURAN), mercaptopurine (PURI- NETHOL), cyclophosphamide (NEOSAR, CYTOXAN), chlorambucil (LEUKERAN), cladribine (LEUSTATIN, MYLINAX), alpha-fetoprotein, etanercept (ENBREL), and 4- benzyloxy-5-((5-undecyl-2H-pyrrol-2-ylidene)methyl)-2,2′-bi-1H-pyrrole (also known as PNU-156804);

[0347] (xx) interferons, including interferon beta-1a (AVONEX, REBIF) and interferon beta- 1b (BETASERON, BETAFERON);

[0348] (xxi) levodopa (or its methyl or ethyl ester), alone or in combination with a DOPA decarboxylase inhibitor (e.g. carbidopa (SINEMET, CARBILEV, PARCOPA, V1512), benserazide (MADOPAR), α-methyldopa, monofluromethyldopa, difluoromethyldopa, brocresine, or m-hydroxybenzylhydrazine);

[0349] (xxii) N-methyl-D-aspartate (NMDA) receptor antagonists, such as memantine (NAMENDA, AXURA, EBIXA), amantadine (SYMMETREL), acamprosate (CAMPRAL), besonprodil (also known as PD-196,860 or CI-1041), ketamine (KETALAR), delucemineAttorney Docket No.: CDPH-003 / 001WO 341287-2404 (also known as NPS 1506), dexanabinol (also known as HU-211), dextromethorphan, dextrorphan, traxoprodil (also known as CP-101,606), himantane, idantadol (also known as V-3381), lancicemine (also known as AR-R 15896), levorphanol (DROMORAN), methadone, (DOLOPHINE), neramexane (also known as MRZ 2 / 579), perzinfotel, phencyclidine, tianeptine (STABLON), dizocilpine (also known as MK-801), ibogaine, voacangine, tiletamine, riluzole (RILUTEK), aptiganel (CERESTAT), gavestinel, and remacimide;

[0350] (xxiii) monoamine oxidase (MAO) inhibitors, such as selegiline (EMSAM), selegiline hydrochloride (I-deprenyl, ELDEPRYL, ZELAPAR), dimethylselegilene, brofaromine, phenelzine (NARDIL), tranylcypromine (PARNATE), moclobemide (AURORIX, MANERIX), befloxatone, safinamide (also known as PNU-151774E), isocarboxazid (MARPLAN), nialamide (NIAMID), rasageline (AZILECT), iproniazide (MARSILID, IPROZID, IPRONID), iproclozide, toloxatone (HUMORYL, PERENUM), bifemelane, desoxypeganine, harmine (also known as telepathine or banasterine), harmaline, linezolid (ZYVOX, ZYVOXID), and pargyline (EUDATIN, SUPIRDYL);

[0351] (xxiv) muscarinic receptor (particularly M1 subtype) agonists, such as bethanechol chloride (DUVOID, URECHOLINE), pilocarpine (SALAGEN), NGX267, arecoline, L- 687306, L-689660, furtrethonium iodide (FURAMON, FURANOL), furtrethonium benzensulfonate, furtrethonium p-toluenesulfonate, McN-A-343, oxotremorine, and carbachol (CARBASTAT, MIOSTAT, CARBOPTIC);

[0352] (xxv) nicotinic receptor agonists, such as epibatidine, ABT-089, ABT-594, AZD- 0328, R4996 (also known as MEM-63908), TC-5619, and EVP-6124;

[0353] (xxvi) neuroprotective drugs such as 2,3,4,9-tetrahydro-1H-carbazol-3-one oxime, AL-108, ACD3480 (also known as TC-1734), bis(4-β-D-glucopyranosyloxybenzyl)-2-β-D- glucopyranosyl-2-isobutyltartrate (also known as dactylorhin B or DHB), xaliproden (XAPRILA), dimeboline hydrochloride (DIMEBON), disufenton (NXY-059, CEROVIVE), arundic acid (ONO-2506, PROGLIA, CEREACT), citicoline (also known as cytidine 5′- diphosphocholine), edaravone (RADICUT), AEOL-10150, AGY-94806 (also known as SA- 450 and Msc-1), granulocyte-colony stimulating factor (AX-200), BAY-387271 (also known as KN-387271), DP-b99, HF-0220 (17-β-hydroxyepiandrosterone), HF-0420 (also known as oligotropin), pyridoxal 5′-phosphate (also known as MC-1), microplasmin, S-18986, piclozotan (also known as SUN-N4057), NP031112, L-seryl-L-methionyl-L-alanyl-L-lysyl- L-glutamyl-glycyl-L-valine, and SUN-N8075;Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0354] (xxvii) norepinephrine (noradrenaline) reuptake inhibitors, such as atomoxetine (STRATTERA), doxepin (APONAL, ADAPIN, SINEQUAN), nortriptyline (AVENTYL, PAMELOR, NORTRILEN), amoxapine (ASENDIN, DEMOLOX, MOXIDIL), reboxetine (EDRONAX, VESTRA), viloxazine (VIVALAN), maprotiline (DEPRILEPT, LUDIOMIL, PSYMION), bupropion (WELLBUTRIN), and radaxafine;

[0355] (xxviii) other PDE9 inhibitors, such as BAY 73-6691 and those disclosed in US Patent Publication Nos US2003 / 0195205, US2004 / 0220186, US2006 / 0111372, and US2006 / 0106035;

[0356] (xxix) other phosphodiesterase (PDE) inhibitors, including (a) PDE1 inhibitors (e.g. vinpocetine (CAVINTON, CERACTIN, INTELECTOL) and those disclosed in U.S. Pat. No. 6,235,742), (b) PDE2 inhibitors (e.g. erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA), BAY 60-7550, and those described in U.S. Pat. No. 6,174,884), (c) PDE4 inhibitors (e.g. rolipram, Ro 20-1724, ibudilast (KETAS), piclamilast (also known as RP73401), CDP840, cilomilast (ARIFLO), roflumilast, tofimilast, oglemilast (also known as GRC 3886), tetomilast (also known as OPC-6535), lirimifast, theophylline (UNIPHYL, THEOLAIR), arofylline (also known as LAS-31025), doxofylline, RPR-122818, or mesembrine), and (d) PDE5 inhibitors (e.g. sildenafil (VIAGRA, REVATIO), tadalafil (CIALIS), vardenafil (LEVITRA, VIVANZA), udenafil, avanafil, dipyridamole (PERSANTINE), E-4010, E-4021, E-8010, zaprinast, PF489791, UK-357903, DA-8159, and those disclosed in International Patent Applications WO05 / 049616, WO06 / 120552, and WO07 / 122466);

[0357] (xxx) quinolines, such as quinine (including its hydrochloride, dihydrochloride, sulfate, bisulfate and gluconate salts), chloroquine, hydroxychloroquine (PLAQUENIL), mefloquine (LARIAM), and amodiaquine (CAMOQUIN, FLAVOQUINE);

[0358] (xxxi) β-secretase inhibitors, such as WY-25105, (+)-phenserine tartrate (POSIPHEN), LSN-2434074 (also known as LY-2434074), PNU-33312, KMI-574, SCH- 745966, Ac-rER (N2-acetyl-D-arginyl-L-arginine), loxistatin (also known as E64d), and CA074Me;

[0359] (xxxii) y-secretase inhibitors, such as LY-411,575, LY-685,458, ELAN-G, ELAN-Z, 4-chloro-N-[2-ethyl-1(S)-(hydroxymethyl)butyl]benzenesulfon-amide;

[0360] (xxxiii) serotonin (5-hydroxytryptamine) 1A (5-HT1A) receptor antagonists, such as spiperone, levo-pindolol, BMY 7378, NAD-299, S(−)-UH-301, NAN 190, WAY 100635, lecozotan (also known as SRA-333);

[0361] (xxxiv) serotonin (5-hydroxytryptamine) 6 (5-HT6) receptor antagonists, such as mianserin (TORVOL, BOLVIDON, NORVAL), methiothepin (also known as metitepine),Attorney Docket No.: CDPH-003 / 001WO 341287-2404 ritanserin, ALX-1161, ALX-1175, MS-245, LY-483518 (also known as SGS518), MS-245, Ro 04-6790, RO 43-68544, Ro 63-0563, RO 65-7199, Ro 65-7674, SB-399885, SB-214111, SB-258510, SB-271046, SB-357134, SB-699929, SB-271046, SB-742457 and PRX-07034;

[0362] (xxxv) serotonin (5-HT) reuptake inhibitors such as alaproclate, citalopram (CELEXA, CIPRAMIL), escitalopram (LEXAPRO, CIPRALEX), clomipramine (ANAFRANIL), duloxetine (CYMBALTA), femoxetine (MALEXIL), fenfluramine (PONDIMIN), norfenfluramine, fluoxetine (PROZAC), fluvoxamine (LUVOX), indalpine, milnacipran (IXEL), paroxetine (PAXIL, SEROXAT), sertraline (ZOLOFT, LUSTRAL), trazodone (DESYREL, MOLIPAXIN), venlafaxine (EFFEXOR), zimelidine (NORMUD, ZELMID), bicifadine, desvenlafaxine (PRISTIQ), brasofensine, and tesofensine;

[0363] (xxxvi) trophic factors, such as nerve growth factor (NGF), basic fibroblast growth factor (bFGF), neurotrophin-3 (NT-3), brain-derived neurotrophic factor (BDNF), and glial- derived neurotrophic factor (GDNF), and agents that stimulate local production of trophic factors, such as propentofylline, idebenone, and AIT-082 (NEOTROFIN); and the like. EXAMPLES

[0364] The Examples below are intended to illustrate particular embodiments of the application and preparations thereto and are not intended to limit the specification, including the claims, in any manner. The compound of Formula (I) is also referred to “Cpd (I)” for short. Example 1: Preclinical Evaluation of a Selective PDE9A Inhibitor

[0365] In recent years, scientific evidence has emerged indicating that PDE9A plays an important role in the biology of heart failure and is upregulated in the cardiac myocytes of patients with heart failure. The compound of Formula (I) (e.g., Compound A) is small molecule inhibitor of PDE9A. An optimal therapeutic agent will demonstrate high affinity for PDE9A and will selectively inhibit PDE9A such that the beneficial effects of cGMP within cardiac myocytes can be potentiated in a sustained manner, without deleterious off-target effects. In Vitro Pharmacology

[0366] The compound of Formula (I) (e.g., Compound A) is an inhibitor of the cGMP- specific PDE9A enzyme with selectivity ≥100x compared to enzymes from other phosphodiesterase gene families. It is a competitive inhibitor of human recombinant PDE9A, with an IC50 of 12 nM (Kleiman et al., J Pharmacol Exp Ther. 2012;341:396-409).Attorney Docket No.: CDPH-003 / 001WO 341287-2404 In Vivo Pharmacology

[0367] The compound of Formula (I) (e.g., Compound A) has been tested in a mouse transaortic constriction (TAC) model of pressure overload induced heart failure. The compound of Formula (I) (e.g., Compound A) significantly reduce left ventricular mass and improve left ventricular function after trans-aortic constriction. Secondary Pharmacodynamics

[0368] The compound of Formula (I) (e.g., Compound A) was evaluated for activity against a broad panel of receptors, enzymes, and ion channels at a single concentration of 10 μM (3955 ng / mL). Less than 50% inhibition of binding or enzyme activity was observed against all targets with the exception of the melatonin 2 receptor (Ki = 3.8 μM) and phosphodiesterase 6 (IC50 = 6.9 μM). These values are well-above the Cmax of the compound of Formula (I) (e.g., Compound A) at the highest dose to be tested in Phase 2 (320 ng / mL), indicating that the compound is unlikely to exhibit significant cross-reactivity with non-PDE9A targets at clinically meaningful exposures. Safety Pharmacology

[0369] The compound of Formula (I) (e.g., Compound A) was assessed in a series of safety pharmacology studies. The oral route of administration was selected for these studies since this is the intended route of clinical administration (note that the anesthetized guinea pig study was conducted by intravenous injection). The cardiovascular assessments were performed in telemetered beagle dogs in cross-over study designs.

[0370] The compound of Formula (I) (e.g., Compound A) was evaluated for potential effects on the pulmonary and central nervous systems (CNS) in male rats at doses up to 300 mg / kg and found to cause mild effects on the CNS (decreased body temperature and locomotor activity) only at the high dose of 300 mg / kg, and there were no biologically significant effects on respiratory parameters (tidal volume, rate of respiration, or minute volume).

[0371] The compound of Formula (I) (e.g., Compound A) was evaluated for effects on the cardiovascular system and found to produce a statistically significant decrease in current amplitude in the hERG assay at concentrations of 10, 30, and 100 μM (equivalent to 3,955; 11,864; and 39,546 ng / mL, respectively), resulting in a calculated 50% maximal inhibitory concentration (IC50) of 55.1 μM (21,790 ng / mL). This corresponds to a margin of ~73x when compared to estimated plasma exposures following a clinical dose of 25 mg BID every 12 hours (q12h). An in vivo cardiovascular screening study in anesthetized guinea pigs indicated that the compound of Formula (I) (e.g., Compound A) has the potential to produce mild cardiovascular effects (e.g. decreased blood pressure and increased heart rate) at plasmaAttorney Docket No.: CDPH-003 / 001WO 341287-2404 concentrations ≥2,333 ng / mL (7x the human total mean maximal concentration (Cmax expected at steady-state of 322.7 ng / mL).

[0372] In the GLP in vivo cardiovascular assessment, male Beagle dogs received doses of 3, 10, or 30 mg / kg of the compound of Formula (I) (e.g., Compound A). There were no biologically relevant changes in blood pressure, left ventricular pressure, or electrocardiogram observed at 3 mg / kg. However, during the first 1-to-5.5-hour interval, there were significant drug-related increases in heart rates (+23 and +24 beats per minute), cardiac contractility (left ventricular maximum +dP / dt, 490 and 626 mm Hg / sec), and corrected QT values (QTc; +9 and +13 msec) at 10 and 30 mg / kg, respectively. Decreases in PR interval (-9 to -10 msec) were considered secondary to the increase in heart rate). No significant cardiovascular effects occurred during later observation periods in the study, indicating that the effects observed at earlier time points were reversible. Pharmacokinetics and Drug Metabolism in Animals Methods of Analysis

[0373] Liquid chromatography with tandem mass spectrometry (LC-MS / MS) was used to determine drug concentrations in plasma samples from pharmacokinetic and serum samples in toxicokinetic studies. Absorption – Single Dose Pharmacokinetics

[0374] After intravenous administration of 1 mg / kg to male rats, the compound of Formula (I) (e.g., Compound A) exhibited a moderate plasma clearance (CL) of 37 mL / min / kg (liver blood flow value 70 mL / min / kg). The steady state volume of distribution (Vss) was 1.9 L / kg, resulting in a short terminal half-life (t½) of 1.5 hours (Table 1). After single oral administration to rats and dogs (1 mg / kg), the oral bioavailability of the compound of Formula (I) (e.g., Compound A) was high with oral bioavailability values of 73% and 91% in rats and dogs, respectively (Table 1). Table 1: Pharmacokinetics in Rats and Dogs == Absolute bioavailability.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Absorption - Repeat Dose Pharmacokinetics (Toxicokinetics)

[0375] The binding of the compound of Formula (I) (e.g., Compound A) to plasma proteins is low (~70 to 90% free) across species. Safety margins may therefore be calculated based on total plasma concentrations.

[0376] Toxicokinetic data were derived from pivotal (definitive) toxicity studies in rats and dogs of up to 6 months and 39 weeks in duration, respectively. These studies were evaluated for sex-related differences in exposure, dose-exposure relationships, and time-dependent changes in systemic exposure to the compound of Formula (I) (e.g., Compound A) in serum.

[0377] The serum toxicokinetics of the compound of Formula (I) (e.g., Compound A) were determined after oral administration to male and female rats at dosages of 10, 50, and 350 mg / kg / day once daily for 3 months. No difference in exposure was observed between male and female animals; therefore, mean data from both male and female animals has been summarized in Table 2. Exposures on Days 1 and 85 were similar, therefore no accumulation of drug was observed over the treatment period.

[0378] Serum toxicokinetics were also determined after oral administration of the compound of Formula (I) (e.g., Compound A) to male and female rats at dosages of 30, 100, and 350 mg / kg / day from postnatal day (PND) 28 to 209 for 6 months. No apparent sex differences in exposure were observed. Mean systemic exposure as assessed by AUC0-24 increased with increasing dose in an approximately dose-proportional manner on PND 208 (Table 2).

[0379] The serum toxicokinetics of the compound of Formula (I) (e.g., Compound A) were determined after once daily oral administration to male and female dogs at doses of 1, 4, and 12 mg / kg / day, for 3 months. No apparent sex differences in exposure were observed between male and female animals and exposure was similar on Days 1 and 88, indicating that there was no apparent drug accumulation. Systemic exposure as assessed by Cmax and area under the concentration time curve (AUC) increased with increasing dose. The mean toxicokinetic parameters for the compound of Formula (I) (e.g., Compound A) are summarized in Table 2.

[0380] Serum toxicokinetics were also determined after oral administration of the compound of Formula (I) (e.g., Compound A) to male and female dogs at dosages of 1, 4, and 12 mg / kg / day for 39 weeks.

[0381] Based on qualitative review of the data, systemic exposure (as assessed by AUC0-24) was higher in males than females at doses of 4 and 12 mg / kg / day; however, variability in exposure was observed. Mean systemic exposure increased with increasing dose on Days 1 and 270. Based on mean AUC values, little or no accumulation occurred between Days 1 and 270 (Table 2).Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Table 2: Summary of Toxicokinetic Data in Nonclinical Species / D5008 0F / 6 5740 4610 26400 23200 14500 11700 66800 58700 a. Based on a molecular weight of 395.5. b. Postnatal day 208 (Rats were dosed daily from postnatal day 28 to postnatal day 208; total 181 doses). c. Non-serial sampling, n=3 / sex / timepoint. AUC0-24 = Area under the concentration time curve from time zero to 24 hours; Cmax = Maximum observed concentration; h = Hour; M = Male; F = Female; N = Number of animals; PO = Oral.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Distribution - Plasma Protein Binding and Blood Partitioning

[0382] The extent of binding of the compound of Formula (I) (e.g., Compound A) to plasma proteins was determined by equilibrium dialysis in mouse, rat, guinea pig, dog, and human plasma at concentrations of 0.1 and 1.0 μg / mL. The compound showed low binding to plasma proteins in all species evaluated. The mean unbound fraction was 0.782, 0.800, 0.684, 0.851, and 0.930 in mouse, rat, guinea pig, dog, and human plasma, respectively. Protein binding was concentration-independent from 0.1 to 1.0 μg / mL.

[0383] Initial studies suggest that the blood partitioning of the compound of Formula (I) (e.g., Compound A) was similar among species examined, with blood-to-plasma ratios ranging from 1.01 to 1.14. Distribution - Tissue Penetration

[0384] Although the compound of Formula (I) (e.g., Compound A) is a substrate for the P- glycoprotein efflux transporter, it had a mean brain-to-plasma ratio of 0.88 and 0.59 in mice and rats, respectively, indicating that this compound was capable of diffusing across the blood-brain barrier. In dogs, brain penetration, as assessed by the cerebrospinal fluid / free plasma ratio, was 0.93, indicating brain penetration in the dog. In Vitro Metabolism

[0385] In preliminary experiments using in vitro systems derived from laboratory animals and humans, the compound of Formula (I) (e.g., Compound A) has been shown to undergo cytochromes P450-mediated metabolism to an N-dealkylated metabolite (M1) and a metabolite arising via opening of the tetrahydropyran ring and subsequent reduction to a di- alcohol (M3). CYP3A4 / 5 were the predominant isoforms responsible for the metabolism, with minor contributions from CYP2D6 and CYP2C8. In Vivo Metabolism

[0386] An assessment of circulating metabolites in humans following oral administration of a single 150 mg dose showed the presence of M1 and M3 as well as a third metabolite arising via hydroxylation of the pyrimidine ring (M2). Comparison of exposures to these three metabolites in humans, rats, and dogs showed that the animals had higher concentrations of M1 and M3, but lower concentrations of the metabolite M2.

[0387] Measurement of M2 in plasma samples from rats following daily oral administration of 300 mg / kg / day and from humans receiving 35 mg BID revealed that humans had a mean average concentration of 74.3 ng / mL whereas this value was 26.9 ng / mL in the rat. Preliminary measurement of this metabolite in urine from humans and rats showed that rats excreted a greater relative amount of M2 than humans (corrected for body weight).Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Excretion

[0388] In preliminary excretion studies in the rat and dog following 1 mg / kg oral administration of the compound of Formula (I) (e.g., Compound A), 4.5% and 8.5% of the administered dose is eliminated unchanged in urine in rat and dog, respectively. Pharmacokinetic Drug Interactions

[0389] DDI risk was determined using steady-state Cmax of 323 ng / mL (total 817 nM; unbound 760 nM) following oral administration of 25 mg BID to humans. This steady-state Cmax was calculated using the observed Day 1 Cmax of 248.2 ng / mL and assuming a 1.3 accumulation ratio in order to derive the steady-state Cmax. Potential for Enzymatic Drug-Drug Interactions

[0390] The potential for the compound of Formula (I) (e.g., Compound A) to inhibit human cytochromes P450 (CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, or CYP3A) was investigated in a preliminary study using human liver microsomes. The compound (30 μM) did not inhibit any activity more than 49%, therefore IC50 values could not be calculated.

[0391] The compound of Formula (I) (e.g., Compound A) (30 μM) after a 30-minute NADPH preincubation, showed no evidence that the compound of Formula (I) (e.g., Compound A) inhibited CYP1A2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, or CYP3A4 (midazolam or testosterone) mediated microsomal turnover in a time-dependent manner. Potential for Drug Transporter Inhibition

[0392] The potential for the compound of Formula (I) (e.g., Compound A) to inhibit various transporters was evaluated in stably expressed mammalian cellular systems. The transporter to be tested is stably expressed in a cultured cell line and the ability of the cell to transport typical molecules specific to the expressed transporter in the presence and absence of the compound is evaluated.

[0393] Inhibition of the efflux transporter P-gp was evaluated in MDCKII-MDR1 cells by comparing the efflux ratio of digoxin (10 μM) in the absence and presence of the compound at concentrations up to 300 μM. Cpd (I) did inhibit P-gp with an IC50 value of 56.9 μM. This indicates that the compound of Formula (I) (e.g., Compound A) has the potential to inhibit P- gp. However, it is unlikely that the compound will inhibit P-gp at clinically relevant concentrations.

[0394] The compound of Formula (I) (e.g., Compound A), at concentrations up to 300 μM, was evaluated for its potential to inhibit the human hepatic uptake transporters OATP1B1 andAttorney Docket No.: CDPH-003 / 001WO 341287-2404 OATP1B3 expressed in human embryonic kidney (HEK) 293 cells. The compound did not inhibit the OATP1B1- and OATP1B3- mediated transport of rosuvastatin (5 μM) up to the highest concentration tested. This indicates that it is unlikely for the compound to inhibit OATP1B1 or OATP1B3 at clinically relevant concentrations.

[0395] The inhibitory potential of the compound of Formula (I) (e.g., Compound A) for the human renal transporters, OAT1, OAT3, MATE1, MATE2K, and OCT2 expressed in HEK293 cells was assessed using [3H]-Para-aminohippurate (2 μM), [3H]-Estrone-3-sulfate (0.2 μM), and [14C]-metformin (10 μM) as probe substrates. The compound did not inhibit OAT1-mediated transport, up to the highest concentration tested (150 μM). Concentration- dependent inhibition of MATE1- and OCT2-mediated transport was observed, with 43.7% and 46.1% inhibition at the highest concentration tested (150 μM). Concentration-dependent inhibition of OAT3- and MATE2K-mediated transport by the compound of Formula (I) (e.g., Compound A) was observed, resulting in estimated IC50 values of 29.5 μM and 5.83 μM, respectively. This indicates that it is unlikely for the compound to inhibit OAT1, MATE1, OCT2, or OAT3 at clinically relevant concentrations, however, the compound of Formula (I) (e.g., Compound A) does have the potential to inhibit MATE2K at clinically relevant concentrations. Pharmacokinetics-Pharmacodynamics

[0396] A TNFα treated mouse model was used as the preclinical pharmacology model to assess the exposure-response relationships of the compound of Formula (I) (e.g., Compound A) given in combination with hydroxyurea on neutrophil adhesion and neutrophil / platelet aggregation in animals. Based on these pharmacodynamic endpoints, a concentration range of 76 to 157 nM (free) corresponding to the minimum concentration (Cmin) at 1 to 10 mg / kg resulted in the statistically significant pharmacodynamic effects. Although the pharmacodynamic readouts are different in heart failure, these data suggest a relationship between target engagement of PDE9A and a pharmacological endpoint. Thus, assuming the potency of the compound is similar between mouse and human, and taking into account the unbound fraction for humans, a dose that maintains a Cmin of approximately 32 to 67 ng / mL (total) is anticipated to be pharmacologically active in humans. Toxicology Single-Dose Toxicity

[0397] Single-dose toxicity of the compound of Formula (I) (e.g., Compound A) was evaluated in male rats at doses of 750, 1000, or 1500 mg / kg, and in male dogs at doses of 10, 30, 100, 150, 200, or 300 mg / kg by oral gavage in an escalating-dose toxicity study. In rats,Attorney Docket No.: CDPH-003 / 001WO 341287-2404 single doses ≥1000 mg / kg were not tolerated, as evidenced by neurologic signs of hypoactivity, tremor, partially closed eyes, and hunched stance that preceded death at ≥1000 mg / kg.

[0398] In dogs, a single dose ≥150 mg / kg in the repeat-dose study was not tolerated based on severe clinical signs including convulsions. The maximum tolerated dose (MTD) of 100 mg / kg in the dog study was associated with decreased activity and emesis. Repeat-Dose Toxicity

[0399] Exploratory and definitive repeat-dose toxicity studies were conducted with the compound of Formula (I) (e.g., Compound A) alone in rats and dogs up to 6 months and 39 weeks, respectively.

[0400] Oral administration of the compound was well tolerated in studies in rats and dogs up to 6 months and 39 weeks in duration, respectively, at doses up to 100 mg / kg / day in rats and at doses up to 12 mg / kg / day in dogs.

[0401] In short-term studies, a dose of 500 mg / kg / day of the compound of Formula (I) (e.g., Compound A) administered for 4 days to male rats was well tolerated and the only findings were minor decreases in sodium and chloride. Following 7 days of dosing, death occurred in male and female rats at doses of 500, 750, and 1000 mg / kg / day. Tremors, convulsions, or general signs of debilitation preceded death and females were more sensitive to the CNS effects of the compound than were males, and all females at ≥750 mg / kg / day were subsequently euthanized. Some of these animals had no premonitory clinical signs on Day 1. Additional neurologic signs included hyperreactivity and tremors at 1000 mg / kg / day and decreased activity, partially closed eyes, and salivation at ≥500 mg / kg / day. Transient hunched posture and recumbency occurred at all doses. Microscopically, there were lesions in the cardiac vasculature in males at ≥500 mg / kg / day (mural degeneration) and in the myocardium in males and females at ≥750 mg / kg / day (myocardial degeneration and inflammation with occasional hemorrhage). Cmax and AUC24 at 500 mg / kg / day were 21,400 ng / mL and 309,000 ng•h / mL, respectively.

[0402] In the dose range- finding study in juvenile rats (postnatal day [PND] 28-PND 42), a dose of 350 mg / kg / day resulted in lower mean body weight gains in males and females following the initiation of dose administration, and consequent lower mean body weights in females only throughout the study. In the 6-month rat study, rats (15 / sex / dose) were administered doses of 30, 100, and 350 mg / kg / day by oral gavage once daily from PND 28 to 209. Test article-related mortality (5 males and 1 female were found dead) and adverse clinical findings (convulsions, hypoactivity, prostrate posture, and a pale and / or cool body)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 were noted in the 350 mg / kg / day group between PND 115 and 208. The histologic findings in the 5 males found dead were considered common, mild, incidental background lesions and the cause of death was not evident microscopically. The cause of death for the female was attributed to chronic progressive nephropathy with tubular dilatation, and a multiloculated cyst noted grossly. There were no test article-related deaths in the pivotal 1- and 3-month studies.

[0403] Daily administration of the compound of Formula (I) (e.g., Compound A) at 10, 50, and 300 mg / kg / day to rats for 1 month was well tolerated, and salivation observed at 300 mg / kg / day was not considered adverse. In the 3-month study, oral doses of 10, 50, and 350 mg / kg / day resulted in hypoactive behavior and squinted eyes at 50 and 350 mg / kg / day in males and 350 mg / kg / day in females. Test article- related mean body weight and mean body weight gain were decreased in males at 350 mg / kg / day. The effects on body weight at 350 mg / kg / day were considered adverse due to the magnitude of the change. After 6 months of dosing, test article-related lower mean body weight gain at 350 mg / kg / day resulted in lower body weight on PND 209 (0.90x control) in males only. There were also test article-related delays in the attainment of balanopreputial separation and vaginal patency observed for 350 mg / kg / day group males and females, respectively. Based on the lack of any other effects in this study that would indicate hormone disruption, these delays were not adverse.

[0404] Test article-related effects on hematology and clinical chemistry parameters were observed in the 1-, 3-, and 6-month rat studies, and were not considered adverse based on the minimal to mild severity of the findings. In the 1-month study, increases in absolute reticulocytes, red cell distribution width (RDW), white blood cell count, and absolute lymphocyte count were noted in females given 300 mg / kg / day. At 3 months, there were no effects on reticulocytes or red blood cell (RBC) parameters; however, there were slight to moderately higher mean values for total leukocyte and lymphocyte counts in the animals given 50 and 350 mg / kg / day, and slightly higher large unstained cell (LUC) counts in females given 350 mg / kg / day. There were no test article-related alterations in hematology parameters in the 6-month rat study.

[0405] At the high doses in the 1- and 3-month pivotal rat studies, clinical chemistry parameter changes included decreased glucose, total protein, albumin, increased glucose, increased blood urea nitrogen (BUN, males); and inorganic phosphorus (P). Other test article- related effects on clinical chemistry parameters in the 1-month study included decreased chloride, and increased cholesterol, alanine aminotransferase (ALT), and calcium. At 3 months, other test article-related changes included slightly higher alkaline phosphatase (ALP)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 in 350 mg / kg / day females, and slightly lower albumin / globulin (A / G) ratio in 10, 50, and 350 mg / kg / day females. One female at 350 mg / kg / day had slightly lower sodium and chloride values. Test article-related changes in urinalysis included a slightly higher urine volume and pH and a lower specific gravity at 50 mg / kg / day in males and 350 mg / kg / day in males and females. Following 6 months of dosing in rats, test article-related changes in clinical chemistry parameters on PND 210 were observed at 350 mg / kg / day and included minimally higher alkaline phosphatase (1.45x control) in females and phosphorus (1.20x and 1.16x controls for males and females, respectively). These findings were not considered adverse due to the small magnitude of difference. There were no test article-related changes in urinalysis parameters.

[0406] A liver change was observed in the 1-month rat study in females. An increase (18%) in absolute liver weight occurred at 300 mg / kg / day, but there was no microscopic correlate associated with the change. In males at the same dose, there was a minimal increase in the number of hyaline droplets in the renal tubular epithelium; this finding is unique to the male rat and not predictive for humans (Hard et al, 1993). Kidney changes were not observed in the 3-month rat study. At a dose of 350 mg / kg / day, a test article-related increase in cytoplasmic vacuolation was observed in the adrenal cortex of males at 350 mg / kg / day and, to a far lesser degree, in females. There were no clinical pathology correlates for this change, and therefore, may represent an exaggerated physiologic response. In males, there was also a slightly increased severity in chronic progressive, nephropathy, which is considered a spontaneous finding in rats. In the 1-month study, cardiomyopathy was noted in 4 males and 1 female given 300 mg / kg / day; however, this finding was also seen and was most severe in 4 vehicle-control males. Cardiomyopathy was not observed in the 3-month study. Cardiomyopathy is a common spontaneous finding in rats, and the incidence / severity of this finding in the control and treated animals was considered within the expected background levels and not considered test article-related (Lewis, 1992). In the 6-month study, test article- related higher (1.14-1.56x control) absolute and relative adrenal gland, heart, liver, and ovary weights were noted in the 350 mg / kg / day group and higher (1.18-1.22x control) adrenal gland weights were noted in the 100 mg / kg / day group males. The higher organ weights did not have microscopic or clinical pathology correlates, and except for the liver weights, were not present at the recovery necropsy. For these reasons, the higher organ weights were not considered adverse. There were no test article-related microscopic findings observed at the initial necropsy or the recovery period necropsy.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0407] The NOAEL for the 6-month rat study was 100 mg / kg / day based on mortality and adverse findings at 350 mg / kg / day. The mean Cmax and AUC0-24 associated with 100 mg / kg / day were 12,200 ng / mL and 141,000 ng•h / mL, respectively.

[0408] In a 10-day study in dogs, test article-related neurologic and gastrointestinal effects were noted and included convulsion in one dog given 100 mg / kg / day. The convulsion was preceded by head shaking / bobbing and cranial vetroflexion. Other signs included walking backwards, unsteady gait, and intermittent tremors. The dose was reduced to 75 mg / kg / day and a majority of the same clinical signs were still observed. Emesis occurred after dosing at ≥30 mg / kg / day while postdose salivation was noted at ≥75 mg / kg / day. The MTD of 30 mg / kg / day was associated with a total Cmax of 20,600 ng / mL and total AUC0-24 of 71,400 ng•h / mL.

[0409] In the pivotal 1-month study in dogs, oral administration of the compound of Formula (I) (e.g., Compound A) was well tolerated at 3 and 10 mg / kg / day for one month since there were no test article-related findings, but doses of 25 and 50 mg / kg / day were not tolerated due to neurological effects (convulsions).

[0410] Originally, a high dose of 50 mg / kg / day had been selected, but this dose was not tolerated and only a single dose was administered. One female at 50 mg / kg / day was euthanized on Day 2 due to convulsions observed on Day 1 that lasted 26 minutes with no premonitory signs. It took two doses of diazepam for the animal to respond. The remaining animals at 50 mg / kg / day had postdose emesis, tremors, excessive salivation, and / or hypoactivity, and dosing was suspended. After a 3-day recovery period, dosing of these animals was reinitiated at 25 mg / kg / day, and a female was replaced. One male at 25 mg / kg / day was euthanized after a convulsion on Day 17 that lasted 20 minutes with no premonitory signs, and tremors. The animal did respond to two doses of diazepam. One male and one female at 25 mg / kg / day had hypoactivity. At ≥10 mg / kg / day, there was a dose- related increase in the incidence of emesis and liquid / nonformed feces and salivation was also noted. There were no effects on any other parameters examined.

[0411] In the pivotal 3-month and 39-week studies in dogs (Studies 6348-574 and 8297047), daily administration of the compound of Formula (I) (e.g., Compound A) at 1, 4, and 12 mg / kg / day was well tolerated. There were no test article-related deaths, adverse findings, or test article-related changes in body weight, food consumption, ophthalmologic, electrocardiographic, heart rate, blood pressure, hematology, coagulation, and urinalysis, macroscopic or microscopic parameters. A test article-related increased occurrence of stoolAttorney Docket No.: CDPH-003 / 001WO 341287-2404 abnormalities (liquid / mucoid / nonformed) was observed at doses of 4 and / or 12 mg / kg / day, but was not considered adverse. Genotoxicity

[0412] The compound of Formula (I) (e.g., Compound A) was assessed in a series of genetic toxicology assays consisting of the microbial reverse mutation, in vitro cytogenetic (human lymphocyte), and in vivo rat micronucleus assays. All in vitro tests were conducted with and without exogenous metabolic activation using concentrations up to those limited by cytotoxicity or insolubility.

[0413] The compound of Formula (I) (e.g., Compound A) was not mutagenic in bacteria, with or without metabolic activation (Table 3). However, the compound of Formula (I) (e.g., Compound A) induced structural chromosome aberrations in human lymphocytes in vitro in the presence of metabolic activation, and this finding was reproducible. The compound of Formula (I) (e.g., Compound A) did not induce chromosomal damage in bone marrow in the 1-month pivotal study in rats at doses producing Cmax and AUC0-24 values significantly exceeding the predicted clinical exposures. Table 3: Genotoxicity Studies Conducted with the compound of Formula (I) (e.g., Compound A) T I I ( e ( e S ( D S e 1 C S ( e ( ( e ( CMicronucleus in Rat Bone Marrowa0, 10, 50, 300 mg / kg / day Negative a. Conducted as part of the 1-month pivotal study in rats. The mean AUC24 and Cmax values at 300 mg / kg / day on Day 25 were 220,000 ng•h / mL and 17,800 ng / mL, respectively.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Reproductive and Developmental Toxicity

[0414] Nonpivotal preliminary oral EFD studies were conducted in pregnant animals to determine doses for the pivotal reproductive and developmental toxicity studies. These nonpivotal rat and rabbit oral dose range-finding studies are not reported herein; however, are included in Table 4. Table 4: Concentrations Associated with Key Responses Key Response(s) Dose AUC 24aCmaxaTotal Total re b N N ↓ (4 ac C N ↓ m M ↓ M p ↓ C ↓ ↑ (4 P ↑ (6 P Si N D st ↓arecumbency, cold to touchAttorney Docket No.: CDPH-003 / 001WO 341287-2404 Key Response(s) Dose AUC 24aCmaxaTotal Total re b Si E E C b E cr v E h in R 7- D or co co tr st ch B cr ab m (1 S n m in h S d fo h p D NAttorney Docket No.: CDPH-003 / 001WO 341287-2404 Key Response(s) Dose AUC 24aCmaxaTotal Total re b N ↓ 1 ↑ E S u sa S w u R 1- H (F gl (F (F w tu E ↓ ce li 3- ↓ ↓ S sq ↑l cr S an Lliver weight (F), ↑cytoplasmic Females)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Key Response(s) Dose AUC 24aCmaxaTotal Total re b v ch 6- N ↑ D re b v ( w w o 1- E sa h S tr m eu S u 3- N L 3 N S(NOEL)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Key Response(s) Dose AUC 24aCmaxaTotal Total re b P N N ↓ co O N N D g w P N ↓ S ↓ S u tr n E NAttorney Docket No.: CDPH-003 / 001WO 341287-2404 Key Response(s) Dose AUC 24aCmaxaTotal Total re b N NALP = Alkaline phosphatase; ALT = Alanine aminotransferase; AUC24 = Area under the concentration-time curve from 0 to 24 hours; BID = Twice daily; BLQ = Below the limit of quantitation; BP = Blood pressure; bpm = Beats per minute; BUN = Blood urea nitrogen; Cmax = Maximum observed calculation; F = Female; HCT = Hematocrit; HGB = Hemoglobin; HR = Heart rate; LUC = Large unstained cells; LV = Left ventricular; MAPD50 = Monophasic action potential duration at 50% repolarization; MAPD90 = Monophasic action potential duration at 90% repolarization (sinus = during basal heart rate; pace = during ventricular pacing at cycle length 200 msec; MCHC = Mean corpuscular hemoglobin concentration; msec = Milliseconds; M = Male; NA = Not applicable; NC = Not calculated; ND =Not determined; NOAEL= No-Observed-Adverse- Effect Level; P = Phosphorus; RBC = Red blood cell; q12h = Every 12 hours; QTc = Corrected QC interval; TK = Toxicokinetic; WBC = White blood cell. a. AUC24 and Cmax values indicate mean serum concentrations (total compound of Formula (I) (e.g., Compound A)). In repeat-dose studies, reported values were obtained near termination. In single-dose studies, AUC and Cmax values were obtained on Day 1. Combined- sex values are used when both sexes were included in the study. b. Exposure margins are based on a human total AUC24 of 3042 ng•h / mL and total Cmax of 322.7 ng / mL at a clinical dose of 25 mg BID (q12h). Total plasma or serum concentrations were used for all calculations. c. Toxicokinetic parameters were not evaluated in this study. Values represent males at Day 1 in the 1-month study in male rats at 50 and 300 mg / kg / day. d. Total dose includes amount of IV loading dose of 0.1 mL / kg / min over 5 minutes followed by 10 minute maintenance infusions of 11.67 μL / kg / min. e. Toxicokinetic parameters were not evaluated at 3 mg / kg in this study. Values represent males at Day 1 in the 1- month study in dogs at 3 mg / kg / day. f. Due to suspected sample switching, AUC24 was recalculated using non-validated software (Pharsight WinNonLin version 5.2). This value is the recalculated value. The original mean AUC24 was 35000 ng•h / mL. g. Not a true Cmax value; mean total plasma concentrations at 6 hours postdose (equivalent safety margin reported in the Cmax exposure column. h. One male died on Day 1 at 20 minutes postdose. i. Animal was euthanized on Day 1 at 4.5 hours postdose following recurring convulsion following subsequent administration of diazepam. j. For the main study and TK study, death occurred 2 / 18 animals at 500 mg / kg / day and 11 / 18 animals at 750 and 1000 mg / kg / day. Surviving female animals at 750 or 1000 mg / kg / day were euthanized on Day 2 due to adverse signs. k. The female dog was euthanized on Day 2 after receiving 100 mg / kg / day on Days 1 and 2. This dose was decreased from 100 to 75 mg / kg / day for the male dog on Day 3. l. Day 10 values from remaining male animal after 8 days at 75 mg / kg / day. m. Value represents concentration 0.26 hours postdose Day 2 at 100 mg / kg / day in female subjected to unscheduled euthanasia. n. AUC and Cmax are listed by sex for each NOAEL. o. Day 1 values. p.5 females were found dead between GD 8 and 17; 3 deaths were considered test article-related, whereas 2 were considered a result of dosing error. q. All females either died or were electively euthanized by GD 8. No noteworthy macroscopic findings were observed and all females were determined to be gravid. r. Not a true Cmax, serum samples collected at 2 hours postdose on GD 8. AUC values are AUC2.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Oral Embryo-Fetal Development in Rats and Rabbits

[0415] In the pivotal EFD study in pregnant female rats (22 / dose), animals were administered doses of 0, 50, 150, or 350 mg / kg / day from gestation days (GD) 6 through 17. There were 3 test article-related deaths at 350 mg / kg / day. Two females in the embryo / fetal development phase and one female in the toxicokinetic phase in the 350 mg / kg / day group were found dead on GD 8. Prior to being found dead, these females had body weight loss and decreased food consumption and one female was noted with red material around the mouth and nose; there were no macroscopic findings. There were two other deaths that were attributed to the dosing procedure and were not considered related to compound administration. Adverse effects on body weight and food consumption were observed at 350 mg / kg / day. There was significantly lower mean maternal body weight gain (0.70x control) and mean maternal body weights (0.96x control) during the overall dosing phase. These differences corresponded to lower mean maternal food consumption observed throughout the dosing phase. Mean gravid uterine weight in this group was significantly lower (0.86x control). There were no adverse effects on body weight and food consumption in rats administered 50 or 150 mg / kg / day.

[0416] Developmental toxicity was manifested in the presence of maternal toxicity at 350 mg / kg / day as lower mean fetal body weights (0.86x control) due in part to lower maternal weights. There were no effects on fetal body weight at 50 and 150 mg / kg / day and no test article-related effects on fetal survival or fetal morphology at any dose. The NOAEL for maternal toxicity was 150 mg / kg / day based on death, and lower body weight and food consumption at 350 mg / kg / day. The NOEL for embryo / fetal development was 150 mg / kg / day based on lower fetal weights at 350 mg / kg / day. The 150 mg / kg / day dose corresponded to an AUC24 of 183,000 ng•h / mL and a Cmax of 15,400 ng / mL on GD 17.

[0417] In the pivotal rabbit EFD study in pregnant female rabbits (20 / dose), there were no test article-related effects on maternal body weights, body weight gains, net body weights and body weight gains, gravid uterine weights, food consumption, or macroscopic examination at any of the doses administered. There were no test article-related effects on fetal weight, fetal survival, or fetal morphology. The NOAEL for maternal toxicity and embryo-fetal development in this study was 50 mg / kg / day (the highest dose tested), which corresponded to a Cmax and AUC24 of 9220 ng / mL and 53,400 ng•h / mL, respectively.

[0418] A preliminary dose range-finding study and a 6-month rat study were conducted in juvenile rats.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Immunotoxicity

[0419] Results in the repeat-dose studies in pre-clinical species showed no evidence of immunosuppression, or inflammatory adverse effects nor any adverse effects on lymphoid tissues. In these studies, the combination of hematologic, organ weight, and macroscopic and histopathologic evaluation provided compelling weight of evidence that the compound of Formula (I) (e.g., Compound A) was not associated with cause for immunotoxicologic concern. Relationship of Findings to Pharmacokinetics

[0420] Exposure to the compound of Formula (I) (e.g., Compound A) in rats and dogs, as defined by Cmax and AUC24, increased with increasing dose over the dose ranges tested. In studies with repeated daily administration, there were no differences when comparing exposure parameters on Day 1 to later days in the study. In the pivotal studies in rats and dogs up to 6 months and 39 weeks in duration, respectively, there were no sex-related differences in exposure at any dose. The threshold serum / plasma concentrations of the compound associated with key responses and exposure margins calculated against these key responses can be found in Table 4. Target Organ Toxicity

[0421] Based on the nonclinical studies conducted with the compound of Formula (I) (e.g., Compound A), the central nervous system and the cardiovascular system have been identified as potential targets in humans. Other findings included effects on the gastrointestinal system in dogs and slight changes in clinical chemistry parameters in rats that were not considered adverse.

[0422] Effects on the CNS were observed in single-dose safety pharmacology studies and in single-dose and repeat-dose studies in rats and dogs following administration of the compound of Formula (I) (e.g., Compound A). Adverse neurological signs in repeat-dose studies were similar to those in single-dose studies. At the high single dose of 300 mg / kg, body temperature and horizontal and vertical locomotor activity were decreased in rats in the neurofunctional safety pharmacology study.

[0423] Single doses of the compound of Formula (I) (e.g., Compound A) in rats at ≥1000 mg / kg were associated with tremor preceding death, hypoactivity, partially closed eyes, hunched stance, salivation, recumbency, and cold to touch. In a repeat-dose study, death at ≥500 mg / kg / day was preceded by tremors, convulsions, or general signs of debilitation. Other animals also were observed with general signs of debilitation including hypoactivity, partially closed eyes, and hunched stance.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0424] Adverse effects on the CNS were observed in dogs in escalating single-dose and repeat-dose studies with the compound of Formula (I) (e.g., Compound A). In the escalating single-dose study, there were convulsions at 150 and 300 mg / kg, necessitating euthanasia at 300 mg / kg. Other neurological signs included: head bobbing, rigid extended limbs, and inability to rise preceding convulsion at 300 mg / kg; head bobbing / shaking, cranial ventroflexion, and walking backwards at 200 mg / kg; hypoactivity at 100 mg / kg; and whole body tremors at 30 mg / kg. In the 10-day study in dogs, convulsions, head bobbing / shaking, unsteady gait, tremors, and salivation were CNS effects associated with a dose of 100 mg / kg / day that was reduced to 75 mg / kg / day following two days of dosing. In the 1-month pivotal study, convulsions resulted in unscheduled euthanasia on Day 2 at 50 mg / kg / day and on Day 17 at the reduced dose of 25 mg / kg / day. Other neurologic findings at 50 mg / kg / day included tremors, hypoactivity, and excessive salivation. There were no CNS effects observed in the 3-month dog study at doses up to 12 mg / kg / day.

[0425] With dogs being the more sensitive nonclinical species, the neurological effects in single-dose escalation and repeat-dose studies occurred at serum concentrations ≥51x the human total mean Cmax of 322.7 ng / mL. Associated AUC24 values in dogs were ≥28x the predicted human total AUC24 of 3042 ng•h / mL at a clinical dose of 25 mg BID (q12h). These neurological effects, particularly convulsions, generally occurred shortly after administration of the compound of Formula (I) (e.g., Compound A) and were associated with the Cmax.

[0426] Effects on the cardiovascular system were observed in the single-dose safety pharmacology study in guinea pigs and dogs, and in repeat-dose studies in rats and dogs administered the compound of Formula (I) (e.g., Compound A).

[0427] In anesthetized guinea pigs, decreased blood pressure and decreased monophasic action potential duration at ≥2333 ng / mL, and increased heart rate at 12,588 ng / mL occurred at compound of Formula (I) (e.g., Compound A) concentrations ≥7x the predicted human total mean Cmax of 322.7 ng / mL at a clinical dose of 25 mg q12h.

[0428] In the 7-day exploratory study in rats with the compound of Formula (I) (e.g., Compound A), there were findings of mural degeneration within a large coronary artery at ≥500 mg / kg / day and ventricular myocardial degeneration with hemorrhage and / or inflammation at ≥750 mg / kg / day. The pathogenesis of the cardiac lesions could not be determined with certainty, although local hemodynamic insufficiency is one possible etiology consistent with vasodilatory effect of treatment, which in turn, would be expected to be associated with transient or sustained reductions in coronary blood flow (Greaves, 2007;Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Histopathology of preclinical toxicology studies. 3rd ed. New York; Elsevier; 2007:299). Doses ≥500 mg / kg are associated with a total AUC24 of 309,000 ng•h / mL, which is approximately 122x the human total AUC24 of 3042 ng•h / mL at a clinical dose of 25 mg BID (q12h).

[0429] In dogs, cardiovascular changes were similar across all studies and were consistent with the known pharmacological effects of the compound of Formula (I) (e.g., Compound A). In the cardiovascular assessments from the safety pharmacology studies, doses of ≥10 mg / kg of the compound of Formula (I) (e.g., Compound A) produced significant findings including increases in HR, cardiac contractility (LV maximum +dP / dt), and QTc. There were no effects on cardiovascular parameters in the 39-week GLP study in dogs with the compound and the total Cmax at doses that elicited cardiovascular effects provided exposure margins ≥9 times the human mean total Cmax of 322.7 ng / mL.

[0430] There were gastrointestinal effects associated with the administration of the compound of Formula (I) (e.g., Compound A) and included dose-related signs of emesis and fecal changes in the pivotal studies in dogs at ≥4 mg / kg / day. There were no associated changes in body weight or food consumption, and there were no histopathological correlates. Based on Cmax and AUC24 at the NOAEL dose of 12 mg / kg / day, safety margins were ≥14x and ≥11x, respectively, relative to the exposure at a projected clinical dose of 25mg BID.

[0431] Test article-related effects on hematology and clinical chemistry parameters were observed in the 1-, 3-, and 6-month rat studies and were not considered adverse based on the minimal to mild severity of the findings. In the 1-month study, increases in absolute reticulocytes RDW, WBC, and lymphocytes were noted in females at 300 mg / kg / day. At 3 months, there were no effects on reticulocytes or RBC parameters; however, there were slight to moderately higher mean values for total leukocyte and lymphocyte counts in the animals given 50 and 350 mg / kg / day and slightly higher LUC counts in females given 350 mg / kg / day. Findings in the 6-month rat study were limited to non-adverse higher alkaline phosphatase and phosphorus at the high dose of 350 mg / kg / day.

[0432] There were no hematologic findings in the pivotal 3-month and 39-week dog studies. Example 2: Effects of the Compound of Formula (I) (e.g., Compound A) In Humans

[0433] Data from a total of eight Phase 1, one Phase 1b, and one Phase 2 study of the compound of Formula (I) (e.g., Compound A) are available. The Phase 1 studies were designed to assess the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics of single and multiple doses of the compound in healthy volunteers (aged 18-85). The Phase 1bAttorney Docket No.: CDPH-003 / 001WO 341287-2404 study included data in safety, pharmacokinetics, and pharmacodynamics of the compound of Formula (I) (e.g., Compound A) co-administered with and without hydroxyurea in subjects with stable sickle cell disease.

[0434] The primary aim of Study 1 was to evaluate the safety, tolerability, PK, and pharmacodynamics of the compound of Formula (I) (e.g., Compound A) after administration of single, escalating, oral doses. The primary objective of Study 2 was to evaluate the safety, tolerability, and PK of multiple doses of the compound of Formula (I) (e.g., Compound A) in healthy elderly subjects. This was an investigator- and subject-blind, sponsor-open, placebo- controlled study. A 5 mg dose was administered every 12 hours (q12h) for 6 days at the time of Study 2 closure and replaced by Study 3. The primary objective of Study 3 was to evaluate the safety, tolerability, and PK of multiple doses of the compound of Formula (I) (e.g., Compound A) in healthy elderly subjects. The first 3 cohorts received the compound of Formula (I) (e.g., Compound A) in an escalating fashion for 7 days. Cohorts 1 (5 mg q12h) and 2 (15 mg q12h) consisted of 8 subjects each (randomized 3:1 drug to placebo). Cohort 3 (35 mg q12h) was dosed in 2 successive groups of 4 subjects each (randomized 3:1 drug to placebo). Cohort 4 (35 mg q12h) consisting of 8 subjects (randomized 3:1 drug to placebo) was treated for 14 days. A total of 32 subjects participated in this study, with 24 receiving treatment with the compound of Formula (I) (e.g., Compound A).

[0435] A total of 17 subjects participated in Study 4, with 15 receiving treatment with the compound of Formula (I) (e.g., Compound A). Fifteen subjects were male and 2 subjects were female, and all subjects were of Japanese descent. A total of 9 subjects were assigned to study treatment in Cohort 1, with 6 subjects each receiving 2 of 3 doses of 3 mg, 10 mg, and 25 mg compound of Formula (I) (e.g., Compound A), and all 9 subjects receiving a single dose of placebo. A total of 8 subjects were assigned to study treatment in Cohort 2, with 6 subjects receiving 25 mg compound q12h and 2 subjects receiving placebo q12h.

[0436] The potential for PK interaction between the compound of Formula (I) (e.g., Compound A) and donepezil, and the safety and tolerability when administered on a background of donepezil therapy were evaluated in 2 small Phase 1 studies: one in healthy volunteers (Study 5), and another in Alzheimer’s disease (AD) subjects (Study 6). Study 6 evaluated the safety and PK of 25 mg dosed q12h for 7 days in these subjects. Five subjects were treated with placebo and 10 subjects were treated with the compound of Formula (I) (e.g., Compound A). Study 5 evaluated the safety and tolerability of the steady-state combination of the compound and donepezil, the effects of steady-state donepezil on the steady-state PK of the compound of Formula (I) (e.g., Compound A), and the effects ofAttorney Docket No.: CDPH-003 / 001WO 341287-2404 steady-state compound of Formula (I) (e.g., Compound A) on the steady-state PK of donepezil. Study 7 evaluated the effect of food on the PK of the compound of Formula (I) (e.g., Compound A) in 10 healthy adult subjects. Subjects were treated once with a dose of 35 mg as an extemporaneously prepared (EP) tablet in the fasted state and once in the fed state, separated by a washout of 1 week.

[0437] A Phase 2 study, Study 8, evaluated the efficacy and safety of 12 weeks of treatment with the compound of Formula (I) (e.g., Compound A) at 25 mg q12h relative to placebo (randomized 1:1). The primary objective of this protocol was to assess the efficacy of the compound of Formula (I) (e.g., Compound A), relative to placebo, on a performance-based measure of cognition (Alzheimer’s Disease Assessment Scale-Cognitive [ADAS-cog]) in subjects with mild to moderate AD. The secondary objectives of this study were to evaluate the effects of the compound of Formula (I) (e.g., Compound A) on other clinically relevant measures including behavior, and clinician-rated global change, to evaluate the safety and tolerability, relative to placebo, and the PK of the compound of Formula (I) (e.g., Compound A).

[0438] Study 9, assessed the safety and tolerability of the compound of Formula (I) (e.g., Compound A) in adult subjects with stable sickle cell disease (SCD) that included subjects with and without hydroxyurea as background therapy; and also aimed to obtain exploratory biomarker data. Two doses (25 and 5 mg, both administered q12h) for 28 days were assessed.

[0439] A Thorough QT / QTc study, Study 10, was conducted. Subjects received in a randomized sequence single oral doses of: 25 mg compound of Formula (I) (e.g., Compound A), 100 mg compound of Formula (I) (e.g., Compound A) (supratherapeutic dose), placebo, and 400 mg moxifloxacin (positive control). At therapeutic and supratherapeutic doses, the compound of Formula (I) (e.g., Compound A) was not associated with QTc interval prolongation that met the threshold of clinical concern based on ICH E14 criteria and therefore satisfied the criteria for a negative Thorough QT / QTc study. The compound of Formula (I) (e.g., Compound A) was generally well tolerated in this subject population.

[0440] Based on these Phase 1 and Phase 2 data, the compound of Formula (I) (e.g., Compound A) was well tolerated. The most frequently reported adverse events associated with treatment were headache, diarrhea, and nausea, and were generally mild in severity. Pharmacokinetics and Product Metabolism in Humans Single Ascending Dose Study

[0441] In the single ascending dose study (Study 1), mean Cmaxvalues ranged from 8.4 to 1198 ng / mL and the area under the plasma concentration-time curve from 0 to the time of lastAttorney Docket No.: CDPH-003 / 001WO 341287-2404 measurement (AUC(0-last)) values ranged from 54.7 to 10,340 ng•h / mL, respectively. The compound of Formula (I) (e.g., Compound A) was rapidly absorbed with median time to maximum plasma concentration (Tmax) of 0.75-1.25 hour postdose.

[0442] The oral volume of distribution (Vz / F) of the compound of Formula (I) (e.g., Compound A) ranged from 128.7 to 713.2 L, suggesting extensive distribution to tissues. Levels in cerebrospinal fluid (CSF) were measured in a cohort of subjects. The CSF / plasma ratio was 0.61 based upon area under the concentration-time curve from 0 to 8 hours (AUC(0-8)). Mean CSF cGMP increased by 246% from baseline with a maximal effect at 6 hours. The pharmacokinetic parameters from study 1 are summarized in Table 5. The compound of Formula (I) (e.g., Compound A) exhibited a biphasic disposition profile, with terminal half-life (t1 / 2) estimates of approximately 19-31 hours for cohorts with adequately characterized biphasic profile (30 mg, 75 mg, and 150 mg). The biphasic nature of the compound was not sufficiently characterized in other cohorts due to samples below the limit of quantitation or inadequate sampling time points. In general, the exposure (Cmax and AUC) increased in a dose-proportional manner. Table 5: Summary of Single Dose Pharmacokinetic Parameters in Healthy Adult Subjectsn AUCinf57.63 (28) 197.5 (25) 734.2 (9) 1533 (14) 2173 (19) 4452 (26) 10420 (29) (ng•h / mL) AUClast54.72 (28) 208.2 (20) 626.0 (24) 1563 (15) 2197 (15) 4418 (27) 10340 (30) (ng•h / mL) Cmax8.436 (16) 28.98 (15) 94.04 (17) 260.9 (14) 297.7 (19) 610.8 (15) 1198 (21) (ng / mL) Tmax(h)1.00 1.00 1.00 (1.00) 0.75 1.25 1.00 1.00(0.50–2.00) (0.50–2.00) (0.50–2.00) (1.25-3.00) (0.98–2.00) (0.50–2.00) t1 / 2(h)5.331 9.124 10.07 31.18 5.019 24.12 19.06(1.345) (4.766) (3.838) (15.38) (0.2875) (9.613) (11.52) CL / F 17350 (39) 14180 (24) 15800 (28) 19570 (15) 18120 (15) 16850 (22) 14390 (26) (mL / h) Vz / F (L)128.7 (12) 271.2 (65) 374.6 (91) 713.2 (50) 223.7 (83) 539.8 (53) 327.9 (69)Geometric mean (%CV) for AUCinf, AUClast, Cmax, CL / F, and Vz / F; arithmetic mean (SD) for t1 / 2; median (range) for Tmax; AUCinf = Area under the plasma concentration-time curve from time zero to infinity; AUClast = Area under the plasma concentration-time curve from 0 to the time of last measurement; CL / F = Oral clearance; Cmax = Maximum plasma concentration; CV = Coefficient of variation; N = Number of subjects; n = Number of subjects contributing to the mean for AUCinf, t1 / 2, CL / F, and Vz / F; SD = Standard deviation; Tmax = Time of occurrence of Cmax; t1 / 2 = Terminal elimination half-life; Vz / F = Terminal volume of distribution.

[0443] Following repeated dosing every 12 hours (q12h) for 7 or 14 days, the exposure of the compound of Formula (I) (e.g., Compound A) appeared to increase in a dose-proportional manner. Absorption of the compound of Formula (I) (e.g., Compound A) was rapid, with median Tmax values of 0.50-1.00 hour post dose. The observed accumulation ratio of theAttorney Docket No.: CDPH-003 / 001WO 341287-2404 compound of Formula (I) (e.g., Compound A) was approximately 1.2-1.5. Steady-state appeared to have been achieved on Day 3. Approximately 17 to 27% of the oral dose was excreted unchanged in urine, indicating that renal excretion is not a major route of elimination for the compound of Formula (I) (e.g., Compound A). Pharmacokinetic parameters are summarized in Table 6. Table 6: Summary of Multiple Dose Pharmacokinetic Parameters In Healthy Elderly Subjects Parameter, Cohort 1 Cohort 2 Cohort 3 Cohort 4 Units 5 mg 15 mg 35 mg 35 mg Study Day 1 N 6 6 5 6 AUCτ, ng•h / mL257.9 (17) 1007 (24) 2132 (24) 2590 (28)Cmax, ng / mL45.83 (20) 174.8 (21) 366.2 (20) 506.1 (35)Tmax, h 1.00 (0.50–1.00) 0.75 (0.50–1.00) 1.00 (1.00–1.00) 0.50 (0.50–1.00) Steady-State N, 6, 6 6, 6 6, 5 6, 6 357.2 (10) 1252 (20) 2602 (17) 3074 (29)61.23 (14) 205.7 (21) 452.2 (17) 484.0 (26)1.00 (0.50–1.00) 0.50 (0.50–1.00) 1.00 (0.50–1.00) 1.00 (0.50–1.00)12.43 (25) 41.92 (30) 75.50 (30) 102.3 (41)4.925 (32) 4.908 (16) 5.990 (28) 4.729 (34) Rac 1.385 (12) 1.244 (8) 1.452 (58) 1.187 (10) t1 / 2, h8.236 (1.567) 10.55 (2.534) 12.87 (2.642) 14.24 (3.673)Urine Pharmacokinetic Parameters N 6 6 6 6 Ae, mg1.340 (31) 3.379 (12) 6.127 (60) 5.884 (35)CLr, mL / min62.53 (33) 44.97 (32) 39.24 (52) 31.90 (47)Food Effect Study

[0444] The effect of food administration on the pharmacokinetics of the compound of Formula (I) (e.g., Compound A) was investigated in Study 7. Following single oral dosing with the EP-tablet formulation of the compound of Formula (I) (e.g., Compound A), mean Cmax was reduced by approximately 20% when administered with a high fat meal. No appreciable change to the AUC was observed. Population Subgroups

[0445] Healthy elderly subjects were studied in the single dose (Study 1) and multiple dose studies (Study 2 and 3). The effect of age on plasma exposure of compound of Formula (I) (e.g., Compound A) was minimal. The pharmacokinetics of the compound was explored in healthy adult and healthy elderly Japanese subjects in Study 4. Pharmacokinetic parameters were similar to those observed in the first in human (FIH) study (Study 1) and multiple dose study (Study 3). Exposure increased in a dose-proportional manner and steady-state was achieved within 3 days of dosing. Dose normalized Cmax values in the Japanese subjects were slightly higher than the previously observed values.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Drug-Drug Interactions

[0446] Studies exploring a potential drug-drug interaction between the compound of Formula (I) (e.g., Compound A) and donepezil were conducted in healthy subjects (Study 5) and in subjects with mild-to-moderate Alzheimer’s disease (Study 6). In both studies, no effect of the compound of Formula (I) (e.g., Compound A) on the pharmacokinetics of donepezil or of donepezil on the pharmacokinetics of the compound of Formula (I) (e.g., Compound A) was observed. A study exploring a potential drug-drug interaction with itraconazole is planned. Human Metabolism

[0447] Preliminary analyses of human plasma samples have found 3 circulating metabolites of the compound of Formula (I) (e.g., Compound A) from the single dose and multiple dose studies, Study 1 and Study 3, respectively. Thorough QT (TQT) Study

[0448] Study 10 was a single-dose study in healthy subjects to evaluate the effect of the compound of Formula (I) (e.g., Compound A) on QT interval. At therapeutic and supratherapeutic doses, the compound was not associated with QTc interval prolongation that met the threshold of clinical concern based on ICH E14 criteria and therefore this study satisfied the criteria for a negative TQT study. The supratherapeutic dose in this study was designed to provide approximately 4-fold margin above the expected steady-state mean maximum exposure following a dose of 25 mg q12h. The adequacy of study sensitivity was confirmed through the effect of moxifloxacin as positive control on the QTc interval. Safety and Efficacy

[0449] The Phase 1 program was designed to assess the safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of single and multiple doses of the compound of Formula (I) (e.g., Compound A) in healthy young adult (aged 18-55 years) and healthy elderly (aged 65-85 years) subjects. One Phase 2 study (Study 8) was designed to assess the efficacy, safety, tolerability, and PK of 12 weeks treatment with the compound of Formula (I) (e.g., Compound A) in subjects with mild to moderate Alzheimer’s disease (AD). In addition, a Phase 1b study was designed to evaluate the safety, PK, and PD of the compound of Formula (I) (e.g., Compound A) in subjects with stable sickle cell disease; and a thorough QT / QTc study was conducted which showed that at therapeutic and supratherapeutic doses, the compound of Formula (I) (e.g., Compound A) was not associated with clinically significant QTc prolongation.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Study 1

[0450] Study 1 was an investigator- and subject-blind, sponsor-open study in healthy young and elderly subjects of ascending oral doses of the compound of Formula (I) (e.g., Compound A). In both the young and elderly cohorts, the study was designed as a 3-period, randomized crossover study with placebo substitution such that each subject was to receive 2 successive dose levels of the compound of Formula (I) (e.g., Compound A) (single doses ranging from 1 to 150 mg) and placebo 1 week apart. An additional cohort of healthy young subjects received a single dose of 40 mg or placebo to assess the effects of the compound of Formula (I) (e.g., Compound A) on CSF cGMP modulation and central nervous system (CNS) penetration. A total of 27 subjects (23 healthy young adult volunteers and 4 healthy elderly volunteers) received single oral doses of the compound of Formula (I) (e.g., Compound A) ranging between 1 and 150 mg, and 2 subjects received a single dose of placebo only in the CSF cohort. The number of subjects receiving treatment at each dose level is shown in Table 7. Table 7: Number of Subjects Receiving Each Treatment in Study 1 G o Y E Y C*Not including young CSF cohort. CSF = Cerebrospinal fluid.

[0451] No deaths, SAEs, or discontinuations due to adverse events (AEs) were reported. There were no severe AEs except in the subjects in the CSF cohort. These included two subjects with headache (1 following 40 mg, 1 following placebo) and 1 subject with neck pain (following 40 mg). A total of 8 AEs, all mild in severity, were reported in the 18 young subjects not included in the CSF cohort: 1 following a 3 mg dose of the compound of Formula (I) (e.g., Compound A), 2 following a 30 mg dose, 4 following a 150 mg dose, and 1 following placebo. No AEs were reported following a dose of 1 mg, 10 mg, or 75 mg. The most frequently reported AEs were nausea (3 subjects: all at the 150 mg dose) and abdominal distension (2 subjects: 1 at the placebo dose, 1 at the 3 mg dose); both AEs were considered by the investigator to be related to study drug.

[0452] A total of 10 AEs were reported in the 4 elderly subjects: 3 following a 30 mg dose, 4 following a 75 mg dose, 1 following a 120 mg dose, and 2 following placebo dosing. TheAttorney Docket No.: CDPH-003 / 001WO 341287-2404 most frequently reported AEs were pain in extremity (3 subjects; 1 subject each at the 30 mg and 75 mg doses and 1 subject following the placebo dose) and arthropod sting (2 events in 1 subject - following the 30 mg and 75 mg doses); all of which were considered by the investigator to be not related to study drug. All AEs in the elderly cohort were mild in severity except for the arthropod stings which were rated as moderate and dry skin rated as moderate in 1 subject following the 120 mg dose.

[0453] A total of 26 AEs were reported in the 7 young subjects comprising the CSF cohort: 12 following a 40 mg dose in 4 subjects, and 14 following placebo dosing in 2 subjects. All of the 12 AEs reported following a dose of 40 mg were considered related to spinal catheterization. Of the

[0454] 14 AEs reported in the CSF cohort following placebo dosing, only 2 events (ear discomfort and feeling hot) were not considered related to spinal catheterization. No AEs were considered related to study drug. No clinically important changes from baseline were observed for laboratory parameters, vital signs measurements, electrocardiogram (ECG) readings, or cardiac telemetry monitoring. Study 2

[0455] Study 2 was a multiple-dose, dose-escalation, investigator- and subject-blind, sponsor-open, placebo-controlled study of the compound of Formula (I) (e.g., Compound A) in healthy elderly subjects. A 5 mg dose or placebo was administered every 12 hours (q12h) for 6 days to 10 subjects at which time the study was closed and replaced by Study 3 (see below).

[0456] There were no subject deaths during the study. One (1) subject in the group receiving the compound of Formula (I) (e.g., Compound A) had an SAE of atrial fibrillation during the study that was not considered to be related to study drug treatment.

[0457] No subject discontinued the study, or had a dose reduction, due to an AE. Nine of 10 subjects (both subjects in the placebo group and 7 of 8 subjects in the group receiving the compound of Formula (I) (e.g., Compound A)) each experienced one or more AEs during this study. Adverse events noted in more than one subject are shown in Table 8. Tabl 8 S mm r f Ad r E nt R rt d b >1 S bj t in St d 2 P S D N PPyrexia 5 1Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Preferred Term Cpd (I), 5 mg q12h; n = 8 Placebo n=2 P C R P A

[0458] Treatment-related AEs included diarrhea, pollakiuria, nausea, and abdominal pain; all of which were mild in intensity. Most of the AEs that were not considered by the investigator as being related to treatment occurred ≥10 days after the start of the study. Several subjects reporting AEs had fever (pyrexia) (n = 6) and cough (n = 6), with some having additional AEs of nausea, fatigue, sore throat, rhinorrhoea, chest congestion, or pneumonia. The investigator felt that a viral illness was the cause of the constellation of symptoms experienced by these subjects. No clinically significant laboratory abnormalities, vital signs, or ECG results were noted during this study. Study 4

[0459] The primary objective of Study 4 was to evaluate the safety and tolerability of the compound of Formula (I) (e.g., Compound A) after oral administration of single and multiple doses to healthy young and elderly Japanese subjects and to characterize the PK of the compound of Formula (I) (e.g., Compound A) after dosing. This was a 3-way crossover study of 3 escalating single doses, and a 1-period study of multiple doses. It was a randomized, investigator- and subject-blind, sponsor-open, placebo-controlled study. A total of 17 subjects participated in Study 4, with 15 receiving treatment with the compound of Formula (I) (e.g., Compound A). All subjects completed the study without any discontinuation.

[0460] There were no deaths, SAEs, withdrawals due to AEs, or temporary discontinuations or dose reductions due to AEs reported. An overview of the number of subjects with AEs is provided in Table 9. The table displays both the number of all-causality AEs and the number of AEs assessed as treatment-related.

[0461] Only 1 subject in Cohort 1 had a laboratory abnormality that met the criteria for potential clinical concern during the study; this subject was on 25 mg of the compound of Formula (I) (e.g., Compound A). In Cohort 2, a total of 3 subjects on 25 mg q12h of the compound of Formula (I) (e.g., Compound A) and 1 subject on placebo had laboratory abnormalities that met the criteria for potential clinical concern during the study. One subject who was on 25 mg q12h of the compound of Formula (I) (e.g., Compound A) in Cohort 2 had a laboratory AE of haematuria. This event was mild in intensity and was considered by the investigator to be related to the study drug. This event did not meet the criteria for potentialAttorney Docket No.: CDPH-003 / 001WO 341287-2404 clinical concern. One subject experienced aspartate aminotransferase (AST) and ALT increases that did not meet the criteria for potential clinical concern, but did exhibit a temporal relationship to study drug administration. Changes from baseline in supine and standing systolic and diastolic blood pressure (BP), pulse rate, and ECG values throughout the study were assessed as not clinically significant. Table 9: Overview of Treatment-Emergent Adverse Events – All-Causality (Treatment-Relate din Study 4) 2Subjects evaluable for AEs 6 6 6 9 6 2 Number of AEs 0 0 1 (0) 1 (0) 6 (4) 1 (1) Subjects with AEs 0 0 1 (0) 1 (0) 2 (2) 1 (1) Subjects with SAEs 0 0 0 0 0 0 Subjects with severe AEs 0 0 0 0 0 0 Subjects who discontinued 0 0 0 0 0 0 due to AEsSubjects with dose 0 0 0 0 0 0 reductions or temporary discontinuations due to AEsIncludes all data collected since the first dose of study drug. Except for the number of AEs, subjects are counted only once per treatment in each row. AEs = Adverse events; q12h = Every 12 hours; SAEs = Serious adverse events. Study 7

[0462] Study 7 was an open-label, randomized, 2-period, 2-treatment, 2-sequence (AB and BA), cross-over, single-dose study in 10 healthy adult male or female subjects, with ages ranging from 18 to 55 years. This study was designed to characterize the effect of food on PK of the compound of Formula (I) (e.g., Compound A) when administered as an extemporaneously prepared (EP) tablet formulation. This was the first clinical study using the EP tablet formulation, which was planned for use in subsequent clinical studies. The 35 mg dose was chosen because it was within the range of well-tolerated single doses in healthy subjects. The expected exposure was below the individual maximum plasma concentration (Cmax) exposure limit of 600 ng / mL (~1 / 7th the observed mean Cmax from the NOAEL dose in the 1-month dog toxicology study), and it was expected to produce significant elevations in CSF cGMP. The PK sampling times were chosen based on the observed PK profile of the oral solution.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0463] A summary overview of treatment-emergent AEs (TEAEs) displaying both all- causality TEAE and treatment-related TEAEs is presented in Table 10. Table 10: Treatment-Emergent Adverse Events Subjects Cpd (I) Cod (I) 35 mg EP Fasted 35 mg EP Fed All-Causality All-causality (Treatment-related) (Treatment-related) Subjects evaluable for AEs 10 (10) 10 (10) Number of AEs 5 (4) 0 (0) Subjects with AEs 2 (2) 0 (0) Subjects with SAEs 0 (0) 0 (0) Subjects with severe AEs 0 (0) 0 (0) Subjects discontinued due to AEs 0 (0) 0 (0) Subjects with dose reduced or temporary 0 (0) 0 (0) discontinuation due to AEs AE = Adverse event; EP = Extemporaneously prepared; SAE = Serious adverse event.

[0464] A summary of the incidence of TEAEs is presented by the System Organ Class (SOC) and Preferred Term (PT) in Table 11. Table 11: Incidence of Treatment-Emergent Adverse Events MedDRA (v12.0) Cpd (I) Cpd (I) SOC and PT 35 mg EP Fasted 35 mg EP Fed (n = 10) (n = 10)All-causality (Treatment-related)All-causality(Treatment-related) Eye Disorders Dry eye 1 (1) 0 (0) Ocular hyperaemia 1 (1) 0 (0) Injury, Poisoning, and Procedural Complications Arthropod bite 1 (0) 0 (0) Nervous System Disorders Dizziness 1 (1) 0 (0) Headache 1 (1) 0 (0) MedDRA (v12.0) = Medical Dictionary for Regulatory Activities (Version 12.0); EP = Extemporaneously prepared; n = Number of subjects; SOC = System organ class; PT = Preferred term..

[0465] There were no vital sign or ECG changes considered clinically significant. One subject in the fasted state experienced a maximum increase of ≥30 mmHg from baseline in standing systolic BP and 1 subject in the fasted state experienced a maximum decrease of ≥30 mmHg from baseline in standing systolic BP. One subject in the fasted state experienced a maximum increase of QTcF interval between 30 and 60 msec from baseline. No subjects showed maximum corrected and uncorrected QT values of ≥500 msec.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0466] In conclusion, a single dose (35 mg) of the compound of Formula (I) (e.g., Compound A) EP-tablet formulation was considered safe and well tolerated in the fed and fasted states of healthy subjects. Study 6

[0467] This study was a double-blind, sponsor-open, randomized, placebo-controlled, parallel arm, multiple-dose study of the compound of Formula (I) (e.g., Compound A) in subjects with mild to moderate AD who were on stable donepezil therapy. The study evaluated the safety and PK of 25 mg of oral compound of Formula (I) (e.g., Compound A) dosed q12h for 7 days.

[0468] There were no deaths, SAEs, withdrawals due to AEs, or temporary discontinuations or dose reductions due to AEs reported in this study. Fifteen AEs were reported for 10 subjects treated with the compound of Formula (I) (e.g., Compound A), q12h plus donepezil QD (13 treatment-related AEs) and no AEs were reported for the 5 subjects treated with placebo q12h plus donepezil QD. Seven of the 15 AEs reported were mild in severity and 8 were moderate. An overview of the number of subjects with AEs, both all-causalities and treatment-related, is provided in Table 12. Table 12: Overview of Treatment-Emergent Adverse Events S S NSSSS to S te As Includes all data collected since the first dose of study drug. Except for the number of AEs, subjects are counted only once per treatment in each row. AE = Adverse event; q12h = Every 12 hours; QD = Once daily; SAE = Serious adverse event.

[0469] No ECG or vital sign abnormalities reported in this study met the criteria for AE reporting. None of the laboratory abnormalities reported met the criteria for AE reporting.

[0470] A summary of the incidence of TEAEs by the SOC and PT is provided in Table 13. Table 13: Overview of Treatment-Emergent Adverse Events in Study 6Attorney Docket No.: CDPH-003 / 001WO 341287-2404 SOC and PTCpd (I) q12h + Donepezil QDPlacebo q12h + Donepezil QD All-causality All-causality (Treatment-related) (Treatment-related) Gastrointestinal disorders 4 (4) 0 Constipation 1 (1) 0 Diarrhea 2 (2) 0 Nausea 2 (2) 0 Vomiting 2 (2) 0 Nervous system disorders 4 (3) 0 Dizziness 1 (1) 0 Headache4 (3) 0Psychiatric disorders 2 (2) 0 Confusional state 1 (1) 0 Nightmare 1 (1) 0 Renal and urinary 1 (0) 0 disordersPollakiuria 1 (0) 0 Includes all data collected since the first dose of study drug. AE = Adverse event; MedDRA = Medical Dictionary of Regulatory Activities; PT = Preferred term; q12h = Every 12 hours; QD = Once daily; SOC = System organ class.

[0471] The results of this study showed that multiple doses of the compound of Formula (I) (e.g., Compound A) administered q12h plus donepezil QD were safe and well tolerated in generally healthy male and female subjects with mild to moderate AD, with no deaths, SAEs, or withdrawals due to AEs reported. Mean donepezil concentrations were similar between subjects treated with the compound of Formula (I) (e.g., Compound A) and those treated with a matching placebo for 7 days. Study 3

[0472] Study 3 was a multiple ascending dose (up to 14 days), investigator- and subject- blind, sponsor-open, placebo-controlled randomized study of the compound of Formula (I) (e.g., Compound A) in healthy elderly subjects. A total of 4 successive cohorts of subjects were treated.

[0473] All subjects completed the study. Changes from baseline in supine and standing systolic and diastolic BP, pulse rate, and ECG values were minimal and not clinically significant throughout the study. Reported TEAEs are shown in Table 14. All were mild in severity except 1 headache assessed as moderate but not related to study medication in 1 subject receiving 15 mg q12h. The most common AEs were diarrhea and headache, which generally were transient in duration ranging from minutes to hours. There were no deaths, SAEs, or discontinuations from the study.

[0474] An overview of the number of subjects with AEs (all-causalities and treatment- related) is provided in Table 14.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Table 14: Overview of Treatment-Emergent Adverse Events (All-Causality and Treatment- Related) (Study 3) Number of Subjects Cpd (I) (q12h) Placebo Total Cohort 1 Cohort 2 Cohort 3a / 3b Cohort 4 5 mg 15 mg 35 mg 35 mg All-Causality (Treatment-Related) Subjects evaluable for 6 6 6 6 8 32 AEs Number of AEs 6 (5) 9 (2) 13 (4) 14 (9) 13 (6) 55 (26) Subjects with AEs 5 (4) 5 (2) 5 (3) 5 (5) 7 (4) 27 (18) Subjects with SAEs 0 0 0 0 0 0 Subjects with severe 0 0 0 0 0 0 AEs Subjects who 0 0 0 0 0 0 discontinued due to AEs Subjects with dose 0 0 0 0 0 0 reductions or temporary discontinuations due toAEsIncludes all data collected since the first dose of study drug. Except for the number of AEs, subjects are counted only once per treatment in each row. AE = Adverse event; q12h = Every 12 hours; SAE = Serious adverse event.

[0475] A summary of the incidence of TEAEs by SOC and PT occurring in ≥2 subjects overall is provided in Table 15. Table 15: Incidence of Treatment-Emergent Adverse Events Occurring in ≥2 Subjects (Study 3) N M S G A C D F M T A N H S DDermatitis 0 1 (0) 0 0 1 (0) If the same subject in a given treatment had more than 1 occurrence in the same preferred term event category, only the mostAttorney Docket No.: CDPH-003 / 001WO 341287-2404 severe occurrence is taken. Includes all data collected since the first dose of study drug. = PT = = 12 =

[0476] The most commonly reported AEs that were considered to be treatment-related were also diarrhea and headache. In addition, no clinically significant cardiovascular or neurological AEs were reported.

[0477] All AEs in this study were considered to be mild in intensity, except for one event of headache that was moderate in intensity and considered to be unrelated to the study drug. Study 5

[0478] This study was a double-blind, sponsor-open, randomized, placebo-controlled, parallel arm, 3-period, multiple-dose study of the compound of Formula (I) (e.g., Compound A) in healthy subjects who were concomitantly receiving donepezil. The study evaluated the effects of steady-state donepezil on the steady-state PK of the compound of Formula (I) (e.g., Compound A) and the effects of steady-state compound on the steady-state PK of donepezil. Multiple doses of the compound of Formula (I) (e.g., Compound A) at 25 mg administered to healthy subjects who were concomitantly receiving donepezil were safe and well-tolerated.

[0479] Five subjects discontinued the study due to AEs: 4 subjects (nausea, headache, and vomiting) during treatment with donepezil 10 mg, and one subject (abnormal behavior) during treatment with donepezil 10 mg plus the compound of Formula (I) (e.g., Compound A) at 25 mg. One severe AE (acute psychosis) was reported during this study and was also considered to be an SAE (reported by one subject after drug washout, assessed as not related to study drug or donepezil). Prior to the SAE of acute psychosis, the subject was reported to have an AE of abnormal behavior which started during treatment with donepezil 10 mg and continued during treatment with donepezil 10 mg plus the compound of Formula (I) (e.g., Compound A). All 5 AEs leading to discontinuation were attributed to donepezil in the opinion of the investigator and all resolved by the end of the reporting period.

[0480] All AEs reported for donepezil 10 mg plus placebo were represented by single occurrences with the exception of nausea (3 subjects) and abdominal discomfort, vomiting, somnolence, and lethargy (2 subjects each). All AEs reported for donepezil 10 mg plus the compound of Formula (I) (e.g., Compound A) were represented by single occurrences with the exception of dizziness (4 subjects), abnormal dreams (3 subjects), and diarrhea and headache (2 subjects each).Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0481] Changes from baseline in supine and standing systolic and diastolic BP, pulse rate, and ECG values throughout the study were not considered to be clinically significant and did not meet the criteria for AE reporting.

[0482] An overview of the number of subjects with AEs (all-causalities and treatment- related) is provided in Table 16. The majority of AEs reported were considered to be mild in severity. A summary of the incidence of TEAEs by the SOC and PT is provided in Table 17. Table 16: Overview of Treatment-Emergent Adverse Events (All-Causality and Treatment- Related) (Study 5) Pl bC d (I)Donepezil Donepezil Donepezil S N S S S S to S o dIncludes all data collected since the first dose of study drug. Except for the number of AEs, subjects are counted only once per treatment in each row. The SAEs were according to the assessment provided by investigator. AE = Adverse event; SAE = Serious adverse event. Table 17: Incidence of Treatment-Emergent Adverse Events a ) A C P E S G D A A C D E F NVomiting 0 0 0 9 (9) 2 (2) 0Attorney Docket No.: CDPH-003 / 001WO 341287-2404 MedDRA (v13.0) SOC Placebo Cpd. (I) 25 Donepezil Donepezi Donepezil a ) A G A C D F F I I U i I P C C I H i N D I C D M N D D H H L P P S P A A A A A H I R S RDisordersAttorney Docket No.: CDPH-003 / 001WO 341287-2404 MedDRA (v13.0) SOC Placebo Cpd. (I) 25 Donepezil Donepezi Donepezil a ) A S e R a D C N O P T S S D C P V FIf the same subject in a given treatment had more than 1 occurrence in the same preferred term event category, only the most severe occurrence is taken. Includes all data collected since the first dose of study drug. Subjects counted only once per treatment in each treatment row. MedDRA = Medical Dictionary of Regulatory Activities; n = Number of subjects Study 8

[0483] This was a multi-center, randomized, double-blind, placebo-controlled, parallel-group Phase 2 study evaluating the efficacy and safety of 12 weeks of treatment with the compound of Formula (I) (e.g., Compound A) compared to placebo. The primary objective of this study was to assess the efficacy of the compound of Formula (I) (e.g., Compound A), relative to placebo, on a performance-based measure of cognition in subjects with mild to moderate AD. The secondary objectives of this study were: to evaluate the effects of the compound of Formula (I) (e.g., Compound A) on other clinically relevant measures including behavior, and clinician-rated global change, to evaluate the safety and tolerability of the compound of Formula (I) (e.g., Compound A), relative to placebo, and to evaluate the PK of the compound of Formula (I) (e.g., Compound A) in subjects with mild to moderate AD.

[0484] The results of this study did not show the compound of Formula (I) (e.g., Compound A) to be more effective compared to placebo on measurements of cognition as measured by the Alzheimer’s Disease Assessment Scale–Cognitive (ADAS-cog), behavior as measured by the Neuropsychiatric Inventory (NPI), and clinician-rated global change as measured by the Clinical Global Impression- Improvement (CGI-I).Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0485] A summary of TEAEs, all-causalities, and treatment-related is presented by treatment in Table 18. The incidence of TEAEs was slightly higher in the group receiving the compound of Formula (I) (e.g., Compound A) (63.7% all-causality; 34.1% treatment-related) than the placebo group (58.0% all-causality; 27.0% treatment-related). Table 18: Treatment-Emergent Adverse Events, All-Causalities and Treatment-Related (Study 8) Number (%) of Subjects All-CausalityNumber ofTreatment-Related Number S N S S S S S ta. After completion of the study and database lock, the sponsor was notified of an additional SAE. One subject receiving the compound of Formula (I) (e.g., Compound A) had an SAE 87 days after the last dose of study drug; the event was not considered to be treatment-related and was not included in this table. b. Subject 10611002 (placebo) discontinued treatment due to an AE on Day 44 and died on Day 98. This subject was included in this table as discontinued treatment due to an AE. Except for the number of AEs, subjects were counted only once per treatment in each row. The SAEs were according to the investigator’s assessment. AE = Adverse event; SAE = Serious adverse event.

[0486] The incidence (in 2% or more subjects in either treatment group) of TEAEs, all- causalities, and treatment-related by MedDRA (version 13.0) SOC and PT is presented by treatment in Table 19. The 2% cut-off was used for presentation in this report in order to provide a more inclusive summary of TEAEs.

[0487] When evaluated by SOC, all-causality AEs in the gastrointestinal disorders class occurred in a higher percentage of subjects in the group receiving the compound of Formula (I) (e.g., Compound A) (19.8%) than in the placebo group (5.0%). These gastrointestinal AEs included abdominal pain, diarrhea, dry mouth, dyspepsia, gastroesophageal reflux disease, intestinal obstruction, nausea, and upper gastrointestinal hemorrhage. No other major treatment differences were noted in incidence of AEs by the SOC. The AEs occurring in 5% or more subjects in either of the treatment group were diarrhea, nausea, nasopharyngitis, and headache. The majority of TEAEs were considered to be mild in severity by the investigator. The incidence of laboratory abnormalities, vital signs abnormalities, and ECG abnormalitiesAttorney Docket No.: CDPH-003 / 001WO 341287-2404 was comparable between the compound of Formula (I) (e.g., Compound A) and placebo groups. There was one report each of BP decrease and BP increase in the group receiving the compound of Formula (I) (e.g., Compound A) which were considered as AEs by the investigator. These AEs were not reported as being associated with dizziness, vertigo, or syncope.

[0488] There was one ECG abnormality that was reported as an AE in a subject treated with the compound of Formula (I) (e.g., Compound A): a right bundle branch block that was not considered clinically significant.

[0489] The majority of TEAEs were considered to be mild in severity by the investigator. Severe AEs that occurred in the group receiving the compound of Formula (I) (e.g., Compound A) included 1 event of agitation, 1 event of intestinal obstruction, 1 event of abdominal pain, 1 event of cerebral hematoma, 1 event of fall, and 1 event of hip fracture. Severe AEs that occurred in the placebo group included 1 event of cardiac arrest, 1 event of respiratory arrest, 1 event of traumatic brain injury, 1 event of chronic hepatic failure, 1 event of hepatic cirrhosis, and 1 event of post-operative wound infection. Table 19: Incidence of Treatment-Emergent Adverse Events by System Organ Class and Preferred Term in 2% or More Subjects in either Treatment Group, All- Causalities, and Treatment-Related S T E V G A D N G a A C I O I B I N Iprocedural complicationsAttorney Docket No.: CDPH-003 / 001WO 341287-2404 All-Causality Treatment-Related S T J I B d H N D H S T P A I R m D S ti H SSubjects were only counted once per treatment for each row. The Medical Dictionary for Regulatory Activities (MedDRA, v13.0) coding was applied. N = number of subjects evaluable for adverse events; n = number of subjects with adverse events.

[0490] Seven subjects permanently discontinued treatment and the study due to AEs: 6 in the treatment group receiving the compound of Formula (I) (e.g., Compound A) and 1 in the placebo group. Three of the 6 subjects treated with the compound of Formula (I) (e.g., Compound A) permanently discontinued treatment and the study due to gastrointestinal disorders AEs (dyspepsia, abdominal pain, and diarrhea) that were considered to be treatment- (Table 19).

[0491] One placebo subject discontinued treatment due to an AE on Day 44 and died on Day 98. This subject was included in Table 22 as ‘died’, and was included in Table 18 as discontinued treatment due to an AE.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Table 20: Permanent Discontinuations Due to Adverse Events Treatment Sex / AgeaAE Start AE PreferredSeveritySAEdb cInvestigator(years) Day Term Causality M / 84 35 Agitation Severe Disease under No studyF / 85 78 Intestinal Severe Other illness - Yes obstruction intestinalCpd (I)obstructionF / 81 2 Dyspepsia Mild Study drug No M / 70 2 Abdominal Severe Study drug No painF / 67 11 Diarrhoea Moderate Study drug No M / 84 35 Hip fracture Severe Other-fall Yes F / 79 69 Pneumonia Moderate Other illness - Yes head injury 60 Traumatic Severe Other illness - Yes Placebo brain injury severe rheumatoidarthritisa. Age at screening. b. Day relative to start of study treatment. First day of study treatment = Day 1. c. The Medical Dictionary for Regulatory Activities (MedDRA, v13.0) coding was applied. d. SAE according to investigator’s assessment. This table includes subjects who discontinued treatment and the study due to AEs. Subject 10611002 (placebo) discontinued treatment due to an AE on Day 44 and died on Day 98. This subject was included in Table 22 as ‘died’, and was included in Table 18 as discontinued treatment due to an AE. AE = Adverse event; SAE = Serious adverse event.

[0492] In the group receiving the compound of Formula (I) (e.g., Compound A), a total of 4 subjects had SAEs (no subjects died), 5 subjects had severe AEs, and 6 subjects discontinued treatment due to 1 or more AEs. After completion of the study and database lock, the sponsor was notified of an additional SAE that was not included in Table 18:

[0493] One subject receiving the compound of Formula (I) (e.g., Compound A) had an SAE of gastric cancer 87 days after the last dose of study drug. This event was not considered to be treatment-related.

[0494] In the placebo group, 4 subjects had SAEs (including 2 subjects who died), 3 subjects had severe AEs, and 2 subjects discontinued treatment due to 1 or more AEs. All SAEs are listed by subject and treatment group in Table 21. Only 1 SAE of syncope in a placebo- treated subject was considered to be treatment-related by the investigator. Table 21: Serious Adverse Events Treatment SAE MedDRA Preferred TermaInvestigator CausalitybIntestinal obstruction Unrelated Cerebral hematoma Unrelated Cpd (I) Hip fracture Unrelated Fall Unrelated Gastric cancer Unrelated Placebo Syncope RelatedAttorney Docket No.: CDPH-003 / 001WO 341287-2404 Cardiac arrest Unrelated Respiratory arrest Unrelated Traumatic brain injury Unrelated Pneumonia Unrelated Hepatic cirrhosis Unrelated Chronic hepatic failure Unrelated Ankle fracture Unrelated Postoperative wound infection Unrelated Fall Unrelated a. MedDRA v13.1 coding applied. b.

[0495] Two (2) placebo-treated subjects and no subjects treated with the compound of Formula (I) (e.g., Compound A) died. A summary of the 2 subjects who died during the study is provided in Table 22. Table 22: Deaths TaPla. Age at onset. b. Day of death was calculated as death date minus first active therapy date plus 1. c. The Medical Dictionary for Regulatory Activities (MedDRA, v13.1) coding was applied. Subject 10611002 (placebo) discontinued treatment due to an adverse event on Day 44 and died on Day 98. This subject was included in this table as died and was included in Table 18 as discontinued due to adverse event. Study 9

[0496] This was a Phase 1b, randomized, double-blind (sponsor open), placebo-controlled study to evaluate the safety, tolerability, pharmacokinetics, and pharmacodynamics of the compound of Formula (I) (e.g., Compound A), coadministered with and without hydroxyurea, in subjects with sickle cell disease.

[0497] Overall, a total of 30 subjects (12 were male and 18 were female) were randomized to the study (7 subjects in the treatment group receiving the compound of Formula (I) (e.g., Compound A) 5 mg BID,16 subjects in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 7 subjects in the placebo group). Of the 30 subjects, 29Attorney Docket No.: CDPH-003 / 001WO 341287-2404 subjects were assigned to and received study treatment, 1 subject in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A) withdrew from the study after randomization but prior to study treatment. Of the 29 subjects who received study treatment, 28 subjects completed the study and 1 subject in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A) discontinued from the study due to an SAE.

[0498] All 29 subjects treated with the compound of Formula (I) (e.g., Compound A) or placebo were included in the safety analysis. Overall, 7 subjects experienced a total of 21 TEAEs after receiving the compound of Formula (I) (e.g., Compound A), 5 mg BID, 13 subjects experienced a total of 56 TEAEs after receiving the compound of Formula (I) (e.g., Compound A), 25 mg BID, and 7 subjects experienced a total of 22 TEAEs after receiving placebo. The majority of the events were mild in severity (15 out of 21 TEAEs in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A), 43 out of 56 TEAEs in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 20 out of 22 TEAEs in the placebo group). SAEs were reported by 3 subjects. One subject in the treatment group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A) discontinued due to an SAE. No subject had dose reductions or temporary discontinuations due to AEs. No deaths occurred. Eighteen out of 56 TEAEs in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A) and 5 out of 22 TEAEs in the placebo group were considered treatment-related by the investigator. No TEAEs were considered treatment-related in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A).

[0499] Treatment-emergent SAEs are summarized in Table 23 (all causality). SAEs were reported by 3 subjects: 2 subjects in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A) and 1 subject in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A). None of these SAEs were considered to be related to study drug by the investigator. Table 23: Treatment-Emergent Serious Adverse Events – All Causality N E N S P BSickle cell anaemia with crisis 1 (14.3%) 1 (6.7%) 0Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Number (%) of Subjects Cpd (I) 5 mg Cpd (I) 25 Placebo H B I P Ab organ class. MedDRA version 19.0 coding dictionary applied

[0500] Treatment-emergent non-serious AEs in >5% of subjects are summarized in Table 24 (all causality) and Table 25 (treatment-related). The system organ classes (SOCs) with the most number of subjects with all causality TEAEs are Nervous System Disorders (57.1% in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A), 60.0% in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 28.6% in the placebo group), and General Disorders and Administration Site Conditions (42.9% in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A), 46.7% in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 57.1% in the placebo group). The most frequently reported TEAEs were headache (57.1% in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A), 33.3% in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 28.6% in the placebo group), and fatigue (28.6% in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A), 40.0% in the group receiving 25 mg BID of the compound of Formula (I) (e.g., Compound A), and 28.6% in the placebo group). Table 24: Treatment-Emergent Non-Serious Adverse Events in >5% of Subjects of Any Treatment Group – All Causality Number (%) of Subjects Cpd (I) 5 mg Cpd (I) 25 mg Placebo BID BIDNumber (%) of Subjects with AEs by SOC and MedDRAaPreferred Term Blood and Lymphatic System Disorders 0 2 (13.3%) 1 (14.3%) Anemia 0 1 (6.7%) 0 Sickle cell anemia with crisis 0 1 (6.7%) 1 (14.3%) Congenital, Familial and Genetic Disorders 0 2 (13.3%) 0 Sickle cell anemia 0 2 (13.3%) 0 Eye Disorders 0 1 (6.7%) 0 Ocular icterus 0 1 (6.7%) 0 Gastrointestinal Disorders 2 (28.6%) 3 (20.0%) 3 (42.9%)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Number (%) of Subjects Cpd (I) 5 mg Cpd (I) 25 mg Placebo BID BID Evaluable for AEs 7 15 7 With AEs 7 (100.0%) 13 (86.7%) 7 (100.0%)SOC and MedDRAaPreferred Term Abdominal distension 1 (14.3%) 0 2 (28.6%) Abdominal pain 0 1 (6.7%) 0 Abdominal pain upper 2 (28.6%) 0 0 Constipation 1 (14.3%) 0 0 Diarrhea 0 1 (6.7%) 0 Dyspepsia 1 (14.3%) 0 0 Nausea 0 1 (6.7%) 2 (28.6%) Sensitivity of teeth 0 1 (6.7%) 0 Vomiting 0 0 1 (14.3%) General Disorders and Administration Site 3 (42.9%) 7 (46.7%) 4 (57.1%) Conditions Chest discomfort 0 1 (6.7%) 0 Fatigue 2 (28.6%) 6 (40.0%) 2 (28.6%) Gait disturbance 0 0 1 (14.3%) Pain 1 (14.3%) 1 (6.7%) 1 (14.3%) Peripheral swelling 0 0 1 (14.3%) Pyrexia 1 (14.3%) 1 (6.7%) 0 Tenderness 0 0 1 (14.3%) Hepatobiliary Disorders 1 (14.3%) 2 (13.3%) 1 (14.3%) Jaundice 0 2 (13.3%) 1 (14.3%) Immune System Disorders 0 0 1 (14.3%) Seasonal allergy 0 0 1 (14.3%) Infections and Infestations 1 (14.3%) 4 (26.7%) 1 (14.3%) Nasopharyngitis 1 (14.3%) 1 (6.7%) 1 (14.3%) Rhinitis 0 1 (6.7%) 0 Upper respiratory tract infection 0 1 (6.7%) 0 Urinary tract infection 0 1 (6.7%) 0 Injury, Poisoning and Procedural 0 0 1 (14.3%) Complications Laceration 0 0 1 (14.3%) Investigations 1 (14.3%) 1 (6.7%) 1 (14.3%) Aspartate aminotransferase increased 1 (14.3%) 0 0 Blood pressure systolic increased 1 (14.3%) 0 0 Neutrophil count decreased 0 0 1 (14.3%) Urine analysis abnormal 0 1 (6.7%) 0 Musculoskeletal and Connective Tissue 1 (14.3%) 5 (33.3%) 1 (14.3%) Disorders Arthralgia 0 2 (13.3%) 0 Back pain 0 2 (13.3%) 1 (14.3%) Bone pain 1 (14.3%) 0 0 Limb discomfort 0 1 (6.7%) 0 Neck pain 0 1 (6.7%) 0 Pain in extremity 0 2 (13.3%) 0Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Number (%) of Subjects Cpd (I) 5 mg Cpd (I) 25 mg Placebo BID BID Evaluable for AEs 7 15 7 With AEs 7 (100.0%) 13 (86.7%) 7 (100.0%)SOC and MedDRAaPreferred Term Nervous System Disorders 4 (57.1%) 9 (60.0%) 2 (28.6%) Dizziness 0 3 (20.0%) 0 Headache 4 (57.1%) 5 (33.3%) 2 (28.6%) Lethargy 0 1 (6.7%) 0 Migraine 0 1 (6.7%) 0 Poor quality sleep 0 1 (6.7%) 0 Somnolence 0 1 (6.7%) 0 Psychiatric Disorders 0 1 (6.7%) 0 Insomnia 0 1 (6.7%) 0 Renal and Urinary Disorders 0 1 (6.7%) 0 Chromaturia 0 1 (6.7%) 0 Reproductive System and Breast Disorders 1 (25.0%) 1 (11.1%) 0 Dysmenorrhea 0 1 (11.1%) 0 Priapism 1 (25.0%) 0 0 Respiratory, Thoracic and Mediastinal 1 (14.3%) 2 (13.3%) 1 (14.3%) DisordersTable 25: Treatment-Emergent Non-Serious Adverse Events in >5% of Subjects of Any Treatment Group – Treatment-Related Number (%) of Subjects Cpd (I) Cpd (I) Placebo 5 mg BID 5 mg BID Evaluable for AE 7 15 7 With AE 0 8 (53.3%) 2 (28.6%) Number (%) of Subjects with AEs by SOC and MedDRAa Preferred Term Gastrointestinal Disorders 0 2 (13.3%) 1 (14.3%) Abdominal distension 0 0 1 (14.3%) Diarrhea 0 1 (6.7%) 0 Nausea 0 1 (6.7%) 1 (14.3%) Sensitivity of teeth 0 1 (6.7%) 0 General Disorders and Administration Site 0 4 (26.7%) 2 (28.6%) Conditions Fatigue 0 4 (26.7%) 2 (28.6%)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Number (%) of Subjects Cpd (I) Cpd (I) Placebo 5 mg BID 5 mg BID Evaluable for AE 7 15 7 With AE 0 8 (53.3%) 2 (28.6%) Musculoskeletal and Connective Tissue 0 1 (6.7%) 0 Disorders Limb discomfort 0 1 (6.7%) 0 Nervous System Disorders 0 6 (40.0%) 1 (14.3%) Dizziness 0 2 (13.3%) 0 Headache 0 4 (26.7%) 1 (14.3%) Poor quality sleep 0 1 (6.7%) 0 Somnolence 0 1 (6.7%) 0 Psychiatric Disorders 0 1 (6.7%) 0 Insomnia 0 1 (6.7%) 0 Skin and Subcutaneous Tissue Disorders 0 1 (6.7%) 0 Pruritus 0 1 (6.7%) 0 Abbreviations: AE=adverse event, BID=twice daily, MedDRA=Medical Dictionary for Regulatory Activities, SOC=system organ class. MedDRA version 19.0 coding dictionary applied.

[0501] All 29 subjects assigned to study treatment had laboratory abnormalities. One subject with preexisting total bilirubin and AST baseline values above the normal range was enrolled in the group receiving 5 mg BID of the compound of Formula (I) (e.g., Compound A). On Study Day 28, the AST value elevated to >3X upper limit of normal (ULN) and the elevation was reported as an AE, which was not considered related to study drug by the investigator.

[0502] There were no clinically significant findings for vital signs or ECG data in this study. Study 10

[0503] This was a Phase 1, single-dose, randomized, 4-treatment, 4-period crossover, double- blind (open label for positive control), sponsor-open study with placebo and positive controls. It was planned to enroll a total of approximately 44 healthy adult subjects to ensure approximately 40 completers. Subjects who withdrew from the study were not replaced.

[0504] Subjects were randomly assigned to 1 of 4 treatment sequences and received single oral doses of 25 mg of the compound of Formula (I) (e.g., Compound A), 100 mg of the compound of Formula (I) (e.g., Compound A), placebo, and 400 mg moxifloxacin in a randomized order (Williams square).

[0505] The primary objective of this study was to assess the effect on QTc of single oral doses of the compound of Formula (I) (e.g., Compound A) relative to placebo. Table 26 provides a summary of the statistical analysis of 25 mg of the compound of Formula (I) (e.g., Compound A) and 100 mg of the compound of Formula (I) (e.g., Compound A) compared to placebo for baseline adjusted QTcF at each postdose time point. Across all time points, 100 mg of the compound of Formula (I) (e.g., Compound A) produced a maximum mean effectAttorney Docket No.: CDPH-003 / 001WO 341287-2404 on QTcF of approximately 5 msec compared to placebo. However, the upper limit of the 2- sided 90% (equivalent to 1-sided 95%) CI at all postdose time points was <10 msec, with a highest value of 7.14 msec, thus satisfying the criteria for a negative thorough QT / QTc study based on ICH E14 guidance. Table 26: Summary of Statistical Analysis of QTcF Between the compound of Formula (I) (e.g., Compound A) and Placebo by Postdose Time Nominal Adjusted Arithmetic Mean Difference e

[0506] The categorical summary of absolute values and maximum increases from baseline for QTcF interval are presented in Table 27. An absolute maximum QTcF interval above 450 but <480 msec was observed in 2 (4.8%) subjects receiving 100 mg of the compound of Formula (I) (e.g., Compound A) and 1 (2.4%) subject receiving moxifloxacin. No subjects had a QTcF interval of ≥480 msec.

[0507] No subjects had a change from baseline in QTcF interval of >30 msec in this study.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Table 27: Categorical Summary of QTcF - Absolute Values and Maximum Increases From Baseline Cpd (I) Cpd (I) MoxifloxacinPlacebo) A IBaseline was defined as the of the means obtained from the measurements taken at -1, -0.5, and 0 hours

[0508] The numbers of subjects with all-causality and treatment-related TEAEs are summarized in Table 28 with the exception of 1 TEAE, all TEAEs were mild in severity. One subject reported a moderate AE of nausea during treatment with 25 mg of the compound of Formula (I) (e.g., Compound A) which resulted in discontinuation. No deaths, severe AEs or SAEs were reported. Two subjects were permanently discontinued from the study due to TEAEs (mild rhabdomyolysis and moderate nausea). Both TEAEs were reported during treatment with 25 mg of the compound of Formula (I) (e.g., Compound A) and neither were considered related to study drug by the investigator. No temporary discontinuations due to AEs were reported. Table 28: Treatment-Emergent Adverse Events - All-Causalities (Treatment-Related)discontinuation due to AEs0 0 0 0Includes data up to 28 days after last dose of study drug. Except for the number of AEs subjects were counted only once per treatment in each row. SAEs - according to the investigator’s assessment. Severity counts are based on the maximum severity or grade of events. MedDRA (Version 19.0) coding dictionary applied. Abbreviations: AE = adverse event; MedDRA = Medical Dictionary for Regulatory Activities; SAE = serious adverse event.

[0509] The incidence of all-causality and treatment-related TEAEs (by system organ class and preferred term) are summarized in Table 29.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0510] The most frequently reported TEAEs were nausea, headache, and dizziness postural; these were reported more frequently following treatment with 25 mg of the compound of Formula (I) (e.g., Compound A) (4, 3 and 2 subjects, respectively) and 100 mg of the compound of Formula (I) (e.g., Compound A) (3, 2 and 2 subjects, respectively) compared to treatment with 400 mg moxifloxacin (1, 2 and 1 subject, respectively) or matching placebo for the compound of Formula (I) (e.g., Compound A) (1, 1, and 0 subject, respectively).

[0511] With the exception of 1 TEAE, all TEAEs were mild in severity. One subject reported a moderate AE of nausea 3 days after receiving treatment with 25 mg of the compound of Formula (I) (e.g., Compound A) which resulted in discontinuation. Table 29: Incidence of Treatment-Emergent Adverse Events - All-Causalities (Treatment- Related) N f i h AEC d IM ifl iM hi Pl o S G A A A D D G N G c P I F L S I A H R N D H H P S P A R U R, Dyspnea 0 1 (1) 0 0Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Number of Subjects With AEsCpd (I)MoxifloxacinMatching Placebo S l 2 1 4 f I N S R T Inclu

[0512] Two subjects discontinued due to TEAEs. Both occurred during treatment with 25 mg of the compound of Formula (I) (e.g., Compound A) and neither were considered related to study drug by the investigator. One subject discontinued due to mild rhabdomyolysis caused by strenuous activity after receiving 25 mg of the compound of Formula (I) (e.g., Compound A) in Period 1. The AE was first noted on Day 7 of Period 1 (6 days after dosing) and resolved 9 days later. Another subject discontinued due to moderate nausea caused by other / unknown reason after receiving 100 mg of the compound of Formula (I) (e.g., Compound A) in Period 1 and 25 mg of the compound of Formula (I) (e.g., Compound A) in Period 2. The AE was first noted on Day 4 of Period 2 (3 days after dosing) and resolved 6 days later.

[0513] Overall, 28 subjects had laboratory abnormalities; there were no observable trends between treatments for any laboratory parameter. Three subjects had changes in LFTs which were considered to be clinically significant; none were considered related to study drug by the investigator.

[0514] None of changes in BP were deemed clinically significant by the investigator. Based on these Phase 1 and Phase 2 study data in healthy volunteers, subjects with mild to moderate AD, and subjects with stable sickle cell disease, the compound of Formula (I) (e.g., Compound A) was well tolerated. The most frequently reported AEs were headache, diarrhea, and nausea, and were generally mild in severity. Overall, no safety findings are apparent which would preclude further development of the compound. Example 3: A Phase 2, Randomized, Double-Blind, Placebo-Controlled Clinical Study to Assess the Effectiveness of the Compound of Formula (I) (e.g., Compound A) in Subjects with Chronic Heart Failure

[0515] As described in Example 2, the compound of Formula (I) (e.g., Compound A) has been generally well tolerated in all clinical studies to date. No major safety concerns wereAttorney Docket No.: CDPH-003 / 001WO 341287-2404 detected for the compound of Formula (I) (e.g., Compound A) to date. No seizures or other major CNS adverse reactions were observed in healthy volunteers or patients. No QTc prolongation was observed at therapeutic and supratherapeutic doses. No other cardiovascular adverse reactions were identified.

[0516] This is a Phase 2, two-part, seamless, randomized, double-blind, placebo-controlled, parallel group clinical study. Part A is a dose ranging study in a group of subjects with CHFrEF. Part B is a proof-of-concept study in subjects with either CHFrEF or CHFpEF. Study Design

[0517] In Part A, eligible subjects with CHFrEF will be randomized to either the compound of Formula (I) (e.g., Compound A) or placebo in a 2:1 ratio. As shown in FIG. 1, subjects will start Period 1 at a dose of the compound of Formula (I) (e.g., Compound A) of 10 mg orally twice daily (BID) or matching placebo. Subjects will remain in Period 1 for 2 weeks. At Day 15, subjects enter Period 2 and are dosed at the compound of Formula (I) (e.g., Compound A) 25 mg orally BID or matching placebo.

[0518] Subjects will continue in Period 2 for 2 weeks. At Day 29, subjects enter Period 3 and continue the compound of Formula (I) (e.g., Compound A) at 25 mg orally BID or matching placebo. Subjects will continue in Period 3 for 8 weeks to complete a total of 12 weeks of the compound of Formula (I) (e.g., Compound A) or matching placebo. Safety and tolerability will be continuously assessed. A post-study visit will occur at Week 16. Part A will evaluate the effectiveness of the compound of Formula (I) (e.g., Compound A) in the overall population of subjects with CHFrEF, and possibly in the 2 subgroups defined by sacubitril / valsartan (Entresto) use (yes, no), depending on the number of subjects in each stratum. Randomization will be stratified on Entresto use (yes vs no).

[0519] The study schematic of Part B is shown in FIG. 2. Part B will initiate enrollment after data review from Part A, where the starting dose for Part B will be chosen based upon all available data, including safety, tolerability, target engagement, and PK, from Part A. Part A subjects may not participate in Part B. The population in Part B will include subjects with CHFrEF and CHFpEF with below-normal systolic function (EF<60%). Subjects will be randomized to either the compound of Formula (I) (e.g., Compound A) or matching placebo in a 1:1 ratio for 12 weeks of treatment. Subjects will be followed with a decentralized format, using a mixture of in-person and remote visits to monitor tolerability and safety. Subjects will be followed for 28 days after the completion of the treatment period for safety. Part B will evaluate the effectiveness of the compound of Formula (I) (e.g., Compound A) in the overall population of subjects with CHF (EF<60%) and in the 8 pre-specified subgroups:Attorney Docket No.: CDPH-003 / 001WO 341287-2404 CHFrEF, CHFpEF, Entresto, no Entresto, CHFrEF with Entresto, CHFrEF without Entresto, CHFpEF with Entresto, and CHFpEF without Entresto. Randomization will be stratified on Entresto use (yes vs no) and on EF (≤ 40% vs > 40%).

[0520] In both Part A and Part B of this study, a subject is considered to have completed the study if he / she has completed the Week 16 visit. The end of the study is defined as the date of the last visit of the last subject in the study. Selection of Doses in the Study

[0521] In Part A, subjects randomized to the compound of Formula (I) (e.g., Compound A) will receive 10 mg of the compound or matching placebo orally BID for 2 weeks and 25 mg of the compound or matching placebo orally BID for 10 weeks. In Part B, subjects randomized to the compound of Formula (I) (e.g., Compound A) will be treated for 12 weeks with 1 dose level of the compound, the selection of which will be based upon all available data, including safety, tolerability, target engagement, and PK, from Part A.

[0522] This clinical study will test doses of the compound of Formula (I) (e.g., Compound A) that have been previously tested safely in humans. The PK profile, safety, and tolerability of the compound of Formula (I) (e.g., Compound A) is well characterized at doses up to 35 mg every 12 hours for 14 days and 150-mg single doses in humans with no specific safety concerns identified. The maximum planned dose in this clinical study is 25 mg BID, which is expected to provide a mean Cmax of ~344 ng / mL.

[0523] While it is not definitively known what the minimum percentage of enzyme inhibition is needed to demonstrate effectiveness of the compound of Formula (I) (e.g., Compound A) in HF, it is hypothesized that an exposure above the half maximal inhibitory concentration (IC50) for PDE-9 will be required. The dose regimens of 10 mg BID and 25 mg BID of the compound of Formula (I) (e.g., Compound A) are expected to achieve exposures above the IC50 for PDE-9, providing margins above the IC50 that thoroughly explore the effect of PDE-9 inhibition while remaining well below a 10-fold margin to the no observed adverse effect level. Objectives (Part A) Primary y To assess the safety and tolerability of the compound of Formula (I) (e.g., Compound A) in subjects with CHFrEF, defined as HF with ejection fraction (EF) ≤40%. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo in plasma cGMP at Week 4 in subjects with CHFrEF.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Secondary y To assess the PK of the compound of Formula (I) (e.g., Compound A) in subjects with CHFrEF. Exploratory y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on N-terminal pro b-type natriuretic peptide (NT-proBNP) at each the compound of Formula (I) (e.g., Compound A) dose and time-point in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) at different dose levels compared to placebo on BNP in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) at different dose levels compared to placebo on plasma and urinary cGMP in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) at different dose levels on the cGMP to NT-proBNP ratio in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) at different dose levels compared to placebo on the cGMP to BNP ratio in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo at Week 12 on the Kansas City Cardiomyopathy Questionnaire-23 (KCCQ-23) scores in subjects with CHFrEF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on the proportion of subjects with ≥5-point improvement in the KCCQ-23 clinical summary score (KCCQ-23-CS) from baseline at Week 12 in subjects with CHF. Objectives (Part B) Primary y To determine whether the compound of Formula (I) (e.g., Compound A) reduces NT- proBNP compared to placebo at Week 12 in subjects with CHF. Secondary y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on BNP at Week 12 in subjects with CHF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP at Week 12 in subjects with CHF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP to NT-proBNP ratio at Week 12 in subjects with CHF.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP to BNP ratio at Week 12 in subjects with CHF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on the KCCQ-23-CS at Week 12 in subjects with CHF. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on the proportion of subjects with ≥5-point improvement in the KCCQ-23-CS from baseline at Week 12 in subjects with CHF. y To determine the relationship between plasma PK exposure of the compound of Formula (I) (e.g., Compound A) and changes in NT-proBNP. y To assess the safety and tolerability of the compound of Formula (I) (e.g., Compound A) in subjects with CHF. Exploratory y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on NT-proBNP in terms of the post baseline mean, at each time-point and over time in subjects with CHF and in all 8 subject subgroups. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on BNP in terms of the post baseline mean, at each time-point and over time in subjects with CHF and in all 8 subject subgroups. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP in terms of the post baseline mean, at each time-point and over time in subjects with CHF and in all 8 subject subgroups. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP to NT-proBNP ratio in terms of the post baseline mean, at each time-point and over time in subjects with CHF and in all 8 subject subgroups. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma cGMP to BNP ratio in terms of the post baseline mean, at each time-point and over time in subjects with CHF and in all 8 subject subgroups. y To assess the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on the KCCQ-23-CS, KCCQ-23 overall summary score (KCCQ-23-OS) and 8 domains in terms of the post baseline mean at Week 12 in subjects with CHF and in all 8 subject subgroups.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 y To determine the relationship between plasma concentration of the compound of Formula (I) (e.g., Compound A) and changes in plasma cGMP in all 8 subject subgroups. Endpoints Primary Efficacy

[0524] Part A: The change from baseline (Day -1) in plasma cGMP at Week 4.

[0525] Part B: The change from baseline (Day 1) in NT-proBNP at Week 12. Primary Safety (Part A only): y AEs y Physical examinations y Vital signs y 12-lead ECGs y Laboratory safety tests (biochemistry, urinalysis, and hematology) Secondary Efficacy (Part B only) y The change from baseline (Day 1) in BNP at Week 12. y The change from baseline (Day 1) in plasma cGMP at Week 12. y The change from baseline (Day 1) in plasma cGMP to NT-proBNP ratio at Week 12. y The change from baseline (Day 1) in plasma cGMP to BNP ratio at Week 12. y The change from baseline (Day 1) in the KCCQ-23-CS at Week 12. y The proportion of subjects with ≥5-point improvement from baseline in the KCCQ- 23-CS at Week 12. Exploratory Efficacy

[0526] Part A: y The change in NT-proBNP from baseline (Day 1) to Week 2, from Week 2 to Week 4, and from Week 2 to Week 12. y The change in BNP from baseline (Day 1) to Week 2, from Week 2 to Week 4, and from Week 2 to Week 12. y The change in plasma, spot urine, and 6-hour urine collection cGMP from baseline (Day - y to Week 2, from Week 2 to Week 4, and from Week 2 to Week 12 (“from Week 2 to Week 12” is NOT applicable for 6-hour urine collection cGMP).Attorney Docket No.: CDPH-003 / 001WO 341287-2404 y The change in cGMP (plasma, spot urine, and 6-hour urine collection) to NT-proBNP ratio from baseline (Day -1) to Week 2, from Week 2 to Week 4, and from Week 2 to Week 12 (“from Week 2 to Week 12” is NOT applicable for 6-hour urine collection cGMP). y The change in cGMP (plasma, spot urine, and 6-hour urine collection) to BNP ratio from baseline (Day -1) to Week 2, from Week 2 to Week 4, and from Week 2 to Week 12 (“from Week 2 to Week 12” is NOT applicable for 6-hour urine collection cGMP). y The change from baseline at Week 12 in the KCCQ-23-CS, KCCQ-23-OS, and 8 domains: physical limitation, symptom stability, symptom frequency, symptom burden, total symptom score, quality of life, self-efficacy, and social limitation. y The proportion of subjects with ≥5-point improvement from baseline in the KCCQ- 23-CS at Week 12.

[0527] Part B: y The post baseline NT-proBNP mean at Weeks 2, 6, and 12. y The post baseline BNP mean at Weeks 2, 6, and 12. y The post baseline plasma cGMP mean at Weeks 2, 6, and 12. y The post baseline plasma cGMP to NT-proBNP ratio mean at Weeks 2, 6, and 12. y The post baseline plasma cGMP to BNP ratio mean at Weeks 2, 6, and 12. y The post baseline mean of the KCCQ-23-OS and 8 domains at Week 12. Secondary Safety (Part B only) y AEs y Physical examinations y Vital signs y 12-lead ECGs y Laboratory safety tests (biochemistry, urinalysis, and hematology Tabl 30 L b t S f t T t B A A A ABUN MCVAttorney Docket No.: CDPH-003 / 001WO 341287-2404 Calcium Platelet count C C D G G I P P S T T U C p B G K L N O PAbbreviations: ALP = alkaline phosphatase; ALT = alanine aminotransferase; AST = aspartate aminotransferase; aPTT: activated partial thromboplastin time; BUN = blood urea nitrogen; CBC = complete blood count; CO2 = carbon dioxide; GGT = gamma glutamyl transferase; INR = international normalized ratio MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; MCV = mean corpuscular volume; PT = prothrombin time; RBC = red blood cell; WBC = white blood cell כ Estimated glomerular filtration rate, as measured by the simplified Modification of Diet in Renal Disease formula, will be derived based on the level of creatinine. Pharmacokinetics

[0528] Part A: The PK endpoints are the following PK parameters of the compound of Formula (I) (e.g., Compound A): y Cmax y Time to reach Cmax (tmax) y AUC0-last y Area under the plasma concentration-time curve over a dosing interval (AUCtau) y Apparent total body clearance (CL / F, except for Day 1)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 y Accumulation ratio for AUCtau [Racc(AUCtau)] and for Cmax [Racc(Cmax)]

[0529] Part B: y Measurement of plasma concentration of the compound of Formula (I) (e.g., Compound A) and correlation with changes in NTproBNP. y Correlation between plasma concentration of the compound of Formula (I) (e.g., Compound A) and changes in plasma cGMP. Estimands

[0530] Table 31 and

[0531] Table 32 address the construction of estimands for the primary objectives for Part A and the primary and key secondary objectives for Part B, respectively. Table 31: Estimands for primary objectives for Part A Objectives Estimands / Endpoints O P T a c ( c ) p ) s y n ) T d t , F C wd heart failure with EF at least one dose of the compound of Formula (I) (e.g.,Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Objectives Estimands / Endpoints ≤ , 2 d y n ) o e dAbbreviations: AE = adverse event; cGMP = cyclic guanosine monophosphate; CHFrEF = chronic heart failure with reduced ejection fraction; EF = ejection fraction; mITT = modified Intention to Treat; SAE = serious adverse event Table 32: Estimands for primary and key secondary objectives for Part B Objectives Estimands / Endpoints O P T P c ( r c C y y nt event is missing at random [MMRM])Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Objectives Estimands / Endpoints e S T c ( c B , s y t n t e e T P c ( c p , i y t nt is missing at random [MMRM])Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Objectives Estimands / Endpoints e m e m T P c ( c p ., p y i t n t e m e m T P c ( c p ., a 2 C t nt is missing at random [MMRM])Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Objectives Estimands / Endpoints e m e m T - c ( c K ., i 2 t n t e m e m T t c ( c p ., ≥ y t t b s nx Subject lost to follow-up (While on treatment strategy)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Objectives Estimands / Endpoints d t , A; CHF = chronic heart failure; CI = confidence interval; EF = ejection fraction; KCCQ-23-CS = Kansas City Cardiomyopathy Questionnaire-23 clinical summary score; LS = least square; mITT = modified Intention to Treat; MMRM = mixed model for repeated measures; NT- proBNP = N-terminal pro b-type natriuretic peptide Inclusion Criteria

[0532] Subjects must satisfy all of the following criteria at the screening visit unless otherwise stated: 1. Able to understand and comply with all study procedures, understand the risks involved in the study, and provide written informed consent. 2. Adult male or female subjects ≥18 years of age, at screening. 3. Subject has evidence in medical history supporting a diagnosis of clinical HF syndrome, New York Heart Association functional class II – III, with the duration of at least 6 months prior to screening. The HF syndrome is defined by documentation of 1 or more of the following: y Dyspnea, paroxysmal nocturnal dyspnea; y Reduced exercise capacity, extended recovery after exercising; y Fatigue; y Peripheral edema (lower leg, ankle); or y Hospitalization or emergency department visit for HF requiring intravenous diuresis within the past 6 months. 4. For Part A: y EF ≤40% by echocardiography at screening. y NT-proBNP level ≥600 pg / ml at screening. Subjects with atrial fibrillation or flutter at screening are required to have an NT-proBNP level of ≥1000 pg / mL at screening. For Part B: y For subjects with EF ≤40%: o EF ≤40% by echocardiography at screening.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 o NT-proBNP level ≥600 pg / ml at screening. Subjects with atrial fibrillation or flutter at screening are required to have an NT-proBNP level of ≥1000 pg / mL at screening. y For subjects with EF >40%: o EF >40% and left atrial enlargement by echocardiography at screening. o NT-proBNP level ≥300 pg / ml at screening. Subjects with atrial fibrillation or flutter at screening are required to have an NT-proBNP level of ≥500 pg / mL at screening. Stable doses of guideline-directed heart failure therapy for a minimum of 4 weeks prior to screening that has been individually optimized according to standard practice guidelines and no addition of guideline-directed heart failure therapy within 3 months of screening. Treatment of HF should include standard of care therapies, such as a beta-blocker and a Renin-Angiotensin-Aldosterone System inhibitor (angiotensin-converting enzyme [ACE]-inhibitor, angiotensin II receptor blocker [ARB]) or combination ARB and neprilysin inhibitor therapy (sacubitril / valsartan [Entresto®]), mineralocorticoid receptor antagonists (MRAs), sodium-glucose transport protein 2 (SGLT-2)-inhibitors, diuretics, or other standard therapies based on practice guidelines and individual subject tolerability. Women of childbearing potential may be considered for enrollment if they are not pregnant, not breastfeeding, do not plan to become pregnant during the study, and are taking appropriate contraception as defined in Inclusion Criterion #7. Women are considered to be of childbearing potential unless they meet at least 1 of the 2 following criteria: a. They have had a hysterectomy, bilateral salpingectomy, or bilateral oophorectomy at a minimum of 1 menstrual cycle prior to signing the informed consent form (ICF); or b. They are post-menopausal: for women ≥55 years of age, defined as ≥1 year since their last menstrual period, or for women <55 years of age, defined as ≥1 year since their last menstrual period and have a follicle-stimulating hormone level in post-menopausal range. Women of childbearing potential must use a double contraception method including a highly effective method of birth control from screening up to 1 month after the last dose. Accepted double contraception methods include the use of intrauterine device orAttorney Docket No.: CDPH-003 / 001WO 341287-2404 hormonal contraception in addition to 1 of the following contraceptive options: (1) condom; or (2) diaphragm or cervical / vault cap. Female subjects who are monogamous with male partners who have had a documented vasectomy is also an accepted method of birth control. 8. Male subjects with female partners of child-bearing potential must agree to use highly effective contraceptive measures from the screening visit through 3 months after the last dose of study medication. Highly effective contraceptive measures include documented vasectomy, abstaining from sexual intercourse, double-barrier method (e.g., male using a condom and female using a diaphragm or cervical cap), or barrier plus hormonal contraception (e.g., male using a condom and female using hormonal contraception or intrauterine device). As there may be a risk of drug being secreted in the ejaculate, male subjects (including men who have had vasectomies) whose partners are currently pregnant should use barrier methods for the duration of the study and for 3 months after the last dose of study medication. Male subjects with female partners that are confirmed as post-menopausal are not required to use contraceptive measures. Exclusion Criteria

[0533] Subjects will be excluded from the study if they satisfy any of the following criteria at the screening visit unless otherwise stated: 1. Documented EF≥60% within 6 months of screening. 2. Recent HF exacerbation defined by hospitalization or requirement for intravenous diuretics within 60 days of screening. 3. Symptomatic hypotension or uncontrolled hypertension (multiple readings with systolic blood pressure >180 mmHg or diastolic blood pressure >110 mmHg) at the screening or baseline visit. 4. History of seizure disorder, or subjects considered to be at increased risk for seizures. 5. Any ECG abnormality that, in the judgment of the investigator, poses a risk to subject safety or impact the validity of study results. 6. Subjects with planned interventions (e.g., percutaneous coronary intervention, devices) etc. occurring during their involvement in this study. 7. Acute coronary syndrome, stroke, transient ischemic attack, cardiac, carotid or other major cardiovascular surgery or carotid angioplasty within 60 days of screening. 8. Subjects with clinical suspicion of infiltrative cardiomyopathy (e.g., amyloid, sarcoid), hypertrophic cardiomyopathy (obstructive or non-obstructive), or HF secondary to severeAttorney Docket No.: CDPH-003 / 001WO 341287-2404 valvular disease, active myocarditis, active pericarditis, or clinically significant congenital heart disease. 9. Prior or planned orthotopic heart transplantation. 10. Presence of or plan for mechanical circulatory support. 11. Chronic treatment with PDE-5 inhibitors (e.g., Viagra, Cialis, Revatio, and Adcirca). 12. Estimated glomerular filtration rate <30 mL / min / 1.73m2as measured by the simplified Modification of Diet in Renal Disease formula at screening. 13. Hepatic impairment at screening defined as alanine aminotransferase / aspartate aminotransferase >3 x upper limit of normal (ULN) and / or total bilirubin >2 x ULN. 14. Concomitant use of a drug that is a known, strong inhibitor or inducer of cytochrome P4503A4 (CYP3A4). 15. Concurrent participation in other clinical studies or receiving other investigational treatments within 30 days of randomization or within 5 half-lives of the other investigational treatment, whichever is longer. 16. In the judgment of the investigator, a history of alcohol or drug abuse which would interfere with the subject’s ability to adhere with protocol requirements. 17. Has evidence of clinically significant cardiovascular, endocrine, gastrointestinal, hematologic, hepatic, immunologic, neurologic, oncologic, pulmonary, psychiatric, or renal disease or any other condition which, in the judgment of the investigator, is not explained by the diagnosis of HF and would jeopardize the safety of the subject or impact the validity of study results. 18. History of malignancy, except subjects who have been disease-free for >5 years prior to screening or subjects whose only malignancy has been successfully treated basal or squamous cell skin carcinoma. 19. Pregnant or nursing (lactating) women, where pregnancy is defined as the state of a female after conception and until the termination of gestation, confirmed by a positive human chorionic gonadotropin laboratory test. 20. Women of childbearing potential, as defined in Inclusion Criterion #6, unless they are using contraception as defined in Inclusion Criterion #7. Discontinuation Criteria Screen Failures

[0534] Screen failures are defined as subjects who consent to participate in the clinical study but are not subsequently entered in the study because of failure to meet the inclusion / exclusion criteria, or any other reason. A minimal set of screen failure informationAttorney Docket No.: CDPH-003 / 001WO 341287-2404 is required to ensure transparent reporting of screen failure subjects to meet the Consolidated Standards of Reporting Trials publishing requirements and to respond to queries from regulatory authorities. Minimal information includes demography, screen failure details, eligibility criteria, and any SAEs. Re-screening

[0535] Subjects who do not meet the criteria for participation in this study may be re- screened once if the reason for the original screen failure is believed not to be present again during the subsequent screening. Re-screened subjects will be assigned a new subject number. Temporary or Permanent Discontinuation of Study Treatment

[0536] In rare instances, it may be necessary for a subject to temporarily interrupt or permanently discontinue the study drug. If a subject requires temporary interruption of the study drug, the study team should consult with the study medical monitor and sponsor. In this study, every attempt will be made to prevent missing data and to obtain complete follow-up of all subjects for the study duration. Investigators will be trained to minimize full withdrawals wherever possible. Subjects who permanently discontinue the study drug are not required to withdraw from the study. These subjects will be encouraged to remain in the study and asked to conduct the remaining study visits as outlined in the protocol. Subject Discontinuation / Withdrawal from the Study

[0537] Subjects may withdraw from the study at any time at his / her own request or may be withdrawn at any time at the discretion of the investigator for safety, behavioral, or administrative reasons. However, every effort should be made to retain the subject in the study. The reasons for subjects not completing the study will be recorded.

[0538] If the subject withdraws consent for disclosure of future information, the sponsor may retain and continue to use any data collected before such a withdrawal of consent. Additionally, subjects may request destruction of any samples taken and not tested. The investigator must document this in the site study records and notify the sponsor or its designee.

[0539] Subjects who withdraw from the study will be asked to complete final study procedures (Visit 9 for Part A and Visit 8 for Part B) as described in the Schedule of Assessments (Table 33 and Table 34). All (S)AEs that are ongoing at the time of withdrawal will be followed up for 28 days, or until resolution or stabilization.

[0540] A subject will be considered lost to follow-up if he / she repeatedly fails to attend scheduled visits and is unable to be contacted by the study site.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0541] Subjects who withdraw from the study will not be replaced. Study Termination

[0542] The sponsor reserves the right to close a study site or terminate the study at any time for any reason at the sole discretion of the sponsor. Study sites will be closed upon study completion. A study site is considered closed when all required documents and study supplies have been collected and a study site closure visit has been performed. Reasons for study termination may include, but are not limited to: y AEs unknown to date (i.e., not previously reported in any similar investigational study drug clinical study with respect to their nature, severity, and / or duration) y Increased frequency and / or severity and / or duration of known, anticipated, or previously reported AEs (this may also apply to AEs defined at check-in as baseline signs and symptoms) y Medical or ethical reasons affecting the continued performance of the study y Difficulties in the recruitment of subjects y Cancelation of drug development.

[0543] The investigator may initiate study site closure at any time, provided there is reasonable cause and sufficient notice is given in advance of the intended termination. Reasons for the early closure of a study site by the sponsor or investigator may include, but are not limited to: y Failure of the investigator to comply with the protocol, the requirements of the institutional review board / ethics committee or local health authorities, the sponsor's procedures, or Good Clinical Practice guidelines y Inadequate recruitment of subjects by the investigator. Study Treatments Treatments Administered

[0544] The investigational product of this study is the compound of Formula (I) (e.g., Compound A) in the form of tablets for oral administration provided in 2 tablet strengths (5 mg and 15 mg). The control of this study is 2 different matching placebo tablets (1 placebo tablet matching the image of the active 5-mg tablet and 1 placebo tablet matching the image of the active 15-mg tablet). The matching placebo tablets are identical in color, size, and shape to the active tablets.

[0545] In Part A of this study, subjects will receive the compound of Formula (I) (e.g., Compound A), 10 mg (two 5-mg tablets) orally BID or matching placebo in Period 1 for 2Attorney Docket No.: CDPH-003 / 001WO 341287-2404 weeks and the compound of Formula (I) (e.g., Compound A), 25 mg (two 5-mg tablets and one 15-mg tablet) orally BID or matching placebo in Periods 2 and 3 for 10 weeks. In Part A, on the days of Visit 5 and Visit 7, subjects will be instructed NOT to take their dose of study drug before the pre-dose blood and urine sampling. Subjects will not take the study drug at Visit 9. They will be instructed to take the study drug up through the day before Visit 9.

[0546] In Part B of this study, subjects will be treated for 12 weeks with 1 dose level of the compound of Formula (I) (e.g., Compound A), the selection of which will be based upon all available data, including safety, tolerability, target engagement, and PK, from Part A, or matching placebo. In Part B, on the days of Visit 4 and Visit 6, subjects will be instructed NOT to take their dose of study drug before the pre-dose blood sampling. Subjects will not take the study drug at Visit 8. They will be instructed to take the study drug through the day before Visit 8.

[0547] In both Part A and Part B of this study, subjects will be instructed to take their study drug orally BID at the same times each day, whenever possible. Preparation, Storage, Handling, and Accountability

[0548] The study drug will be supplied by the sponsor or its designee. The packaging of the 10 mg the compound of Formula (I) (e.g., Compound A) and matching placebo dose will be clearly differentiated from that of the 25 mg the compound of Formula (I) (e.g., Compound A) and matching placebo dose. However, for each dose level, the compound of Formula (I) (e.g., Compound A) and its matching placebo are labeled in a double-blind fashion.

[0549] The investigator or his / her designee must confirm appropriate temperature conditions have been maintained during transit for all study drug received and any discrepancies are reported and resolved before the use of the study drug.

[0550] All study drug must be maintained at room temperature in a locked, secure, environmentally controlled, and monitored (manual or automated) location at the site with access limited to the investigator and authorized site staff.

[0551] Only subjects enrolled in the study may receive the study drug. At the study visits in Parts A or B of this study, as summarized in the Schedule of Assessments (Table 33 and Table 34), site pharmacists will be blinded to treatment allocation and will dispense a sufficient quantity of the compound of Formula (I) (e.g., Compound A) or matching placebo tablets for the subject to self-administer on an out-patient basis. Subjects will be instructed to follow the provided instructions to take the study drug.

[0552] At the study visits in both Part A and Part B of this study, as summarized in the Schedule of Assessments (Table 33 and Table 34), subjects will be asked to return all usedAttorney Docket No.: CDPH-003 / 001WO 341287-2404 and unused drug supply containers. Returned study drug should not be re-dispensed to the subjects. Method of Treatment Assignment

[0553] All subjects will be centrally randomized using an interactive web-based response system (IWRS). Before the study is initiated, the log-in information and directions for the IWRS will be provided to each site. On Day -1 of Part A of this study, eligible subjects will be randomly assigned to either the compound of Formula (I) (e.g., Compound A) or placebo in a 2:1 ratio using the IWRS. On Day 1 of Part B of this study, eligible subjects will be randomly assigned to either the compound of Formula (I) (e.g., Compound A) or placebo in a 1:1 ratio using the IWRS. Randomization will be stratified in Part A on Entresto use (yes vs no) and in Part B on Entresto use (yes vs no) and on EF (≤ 40% vs > 40%). In Part B, capping measures will be applied to ensure adequate sample sizes for each subgroup to assess the study objectives. The investigator or his / her designee will obtain the randomization number assigned to a subject by accessing the IWRS after confirming that the subject meets all inclusion criteria and does not meet any of the exclusion criteria.

[0554] No dose modification other than the protocol-specified dose escalation will be permitted. Blinding and Blind-Breaking Procedures

[0555] This is a double-blind study. On Day -1 of Part A, eligible subjects will be randomly assigned to either the compound of Formula (I) (e.g., Compound A) or placebo in a 2:1 ratio. On Day 1 of Part B, eligible subjects will be randomly assigned to either the compound of Formula (I) (e.g., Compound A) or placebo in a 1:1 ratio. Both investigators and subjects will remain blinded to each subject’s assigned study drug throughout the course of the study. The following controls will be employed to maintain the double-blind status of the study: y For each dose, the compound of Formula (I) (e.g., Compound A) tablets and their matching placebo tablets will be supplied in identical packaging and will be identical in color, smell, taste, and appearance. y The investigator and other members of staff involved with the study will remain blinded to the treatment randomization code. y The site pharmacists responsible for the dispensation of the study drug are blinded to treatment allocation.

[0556] If it becomes necessary to break the code during the study, the date, time, and reason will be recorded in the subject’s source data. Under such circumstances, subjects will beAttorney Docket No.: CDPH-003 / 001WO 341287-2404 withdrawn from the study; dosing of the compound of Formula (I) (e.g., Compound A) or matching placebo will be stopped, and end of treatment procedures will be conducted with documented AE related to the unblinding reason.

[0557] The IWRS will be programmed with blind-breaking instructions. The study blind may be broken if, in the opinion of the investigator, it is in the subject’s best interest to know the study drug assignment. If possible, the sponsor must be notified before the blind is broken unless identification of the study drug is required for a medical emergency in which the knowledge of the specific blinded study drug will affect the immediate management of the subject’s condition (e.g., antidote is available). In this case, the sponsor must be notified within 24 hours after breaking the blind. The date and reason that the blind was broken must be recorded in the source documentation and electronic case report form (eCRF), as applicable.

[0558] In the event of a Quality Assurance audit, the auditor(s) will be allowed access to unblinded study treatment records, if any, at the site(s) to verify that randomization / dispensing has been done accurately. Treatment Compliance

[0559] The study drug will be dispensed at the study visits in Parts A and B of this study, as summarized in the Schedule of Assessments (Table 33 and Table 34). Subjects will be instructed to follow the provided instructions to take the study drug.

[0560] At the study visits in Parts A and B of this study, as summarized in the Schedule of Assessments (Table 33 and Table 34), subjects will be asked to return all used and unused drug supply containers. Accountability and subject compliance will be assessed by maintaining adequate drug dispensing and returning records.

[0561] This is a Phase 2, two-part, seamless, randomized, double-blind, placebo-controlled, parallel group clinical study. Part A is a dose ranging study in a group of subjects with CHFrEF. Part B is a proof-of-concept study in subjects with either CHFrEF or CHFpEF.4042- o) ) )s7et 3161 ya8 1k d20143O W100 / 300-HPDC:.oNtekcoDyenrottA4oy amde ar D1(±(141A3y1aX X X X X X trD a P r1- o f2yX satnD eb.m1ssgote 5 )ss 1nin1e-2- yyX X X X X X X X X X X Aeafo r aD ec DlS (udtiesi hicVa.sc1u y.1tycf.g,.1n TStlccirotsenae.11GPoiPt,:n x3ede / hnai3mesrps ai trhly ncgesnornlitC- aETNa nBzioiTtTPl seemtrn oeisseutirgo acnidmap dungenc. . g11isalcinsidmaNPelmuNamaB moldu a(gle )bs oas f nl c mat e ri tHat y a - rsa n a n RTAnIoccnIneoDadMeeSrr spUetSiFVhPxe21eoSrplPaRoCapNI404) )2s-78201143O W100 / 300- 9HPDC:.oNtekco 8DyenrottA765oe3 )ty4 oy amdX X eaX X r D1(±(1k, ,53y .-1, rh 6t,s)aX X X X X1 ere 2 rop es k.1-1er , reh 6ts)12,rop es 1D Xp( sod ,rhh 3rhodXp( sod ,rhh 3rhod1- k1 ,rh 62y .a X X1,r,Xh2 r)r0, h hD(rh 3b.1got5 )21nin1- -ey yX X X ea arc DD S(tis ji.Va1.1nyati.n 1noi ytioit gtelyaelss)teyrsmso t gtali alnit sdna stn p ssp aneurrtb esatac r ioitmaamaPm sartd is on ne cmo acs g sdudMt eessobyt ydi g uoeaef umur cs ercio E nord)oGceggao de olrKolr ragALastSdaDcaD ACmBofP( Boft(ne404) ) 32s 2fo ni -78201143O W100 / 300- 9HPDC:.oNtekco 8DyenrottA7ya e dX Xrpya e dDW3X Xuirhe(+(ta;nep ms l rnc sit eedopfmreocessatisdotero)e) ep omru yn iredn eb6t7 yyo1 at-ro ir thtailairt,m y.pla,etllia d bgn big.auarw er1(D±=( Pniepgil e(rt ystNtBal yut- essnro nbere e t euc m arme5)12 )s ;yem mit orsna aipris- pne tsdse us5ykaeeadX XmntelX Xn lc an.otmu ahrat ,) e2itil ll ide 5cri o ci m DW(±(sapofmrief1-c n l ntic yn eov tsgiso)ob=- NepaDee ae) h ai la4tyo3y a mH or SF = sPesi nsoeroo dtihdnVm.e a d hr D1t; 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[0562] Any medication or vaccine (including over-the-counter or prescription medicines, vitamins, and / or herbal supplements) that the subject is receiving at the time of ICF signing onwards must be recorded along with: y reason for use y dates of administration including start and end dates y dosage information including dose and frequency

[0563] The medical monitor should be contacted if there are any questions regarding concomitant or prior therapy.

[0564] Any medications, excluding those prohibited as described below, may be given concomitantly as needed for the subject’s welfare for treatment of comorbid conditions or potential AEs. At each visit in this study, the study staff will review use of concomitant medications with the subject and will record entries in the eCRF. Prohibited Concomitant Therapy

[0565] Subjects may not be on the following drugs from the time of ICF signing onwards: y PDE-5 inhibitors (e.g., Viagra, Cialis, Revatio, and Adcirca) y Known strong inhibitors or inducers of CYP3A4. Study Assessments And Procedures

[0566] Study procedures and their timing are summarized in the Schedule of Assessments (Table 33 and Table 34). Adherence to the study design requirements, including those specified in the Schedule of Assessments (Table 33 and Table 34), is essential and required for study conduct. All screening evaluations must be completed and reviewed to confirm that potential subjects meet all eligibility criteria. The investigator will maintain a screening log to record details of all subjects screened and to confirm eligibility or record reasons for screening failure, as applicable.

[0567] All laboratory tests in this study will be performed at a central laboratory. The maximum amount of blood collected from each subject over the duration of the study will be in line with local regulations. Repeat or unscheduled samples may be taken for safety reasons or for technical issues with the samples. Efficacy Assessments

[0568] The NT-proBNP will be assessed at the time-points specified in the Schedule of Assessments (Table 33 and Table 34).Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0569] The KCCQ-23 scores, including the KCCQ-23-CS, KCCQ-23 KCCQ-23-OS, and 8 domains (physical limitation, symptom stability, symptom frequency, symptom burden, total symptom score, quality of life, self-efficacy, and social limitation), will be assessed electronically at the time-points specified in the Schedule of Assessments (Table 33 and Table 34).

[0570] The BNP will be assessed at the time-points specified in the Schedule of Assessments (Table 33 and Table 34).

[0571] Plasma cGMP will be assessed at the time-points specified in the Schedule of Assessments (Table 33 and Table 34).

[0572] In Part A of this study only, urinary cGMP (6-hour urine collection and spot urine) will be assessed at the time-points specified in the Schedule of Assessments (Table 33 and Table 34). Safety and Tolerability Assessments

[0573] AE definitions and assignment of severity and causality are detailed below. AEs will be elicited from the subject (or, when appropriate, from a caregiver, surrogate, or the subject’s legally authorized representative) by the study site staff using a non-leading question such as “How are you feeling today?” or “Have you had any health concerns since your last visit?” Subjects will be encouraged to voluntarily report AEs occurring at any time during the study. AEs will be recorded in the eCRF.

[0574] If a female subject becomes pregnant during the study, the study drug must be discontinued immediately. During the study, pregnancy events should be reported and will be followed until conclusion.

[0575] At the time-points specified in the Schedule of Assessments (Table 33 and Table 34) and at all unscheduled visits as applicable, blood and urine samples will be collected for laboratory safety tests, including hematology, biochemistry, coagulation, and urinalysis. Clinically significant laboratory abnormalities, in the opinion of the investigator, should be reported as AEs.

[0576] Vital signs, including heart rate, blood pressure (systolic and diastolic), respiratory rate, and temperature will be measured at the time-points specified in the Schedule of Assessments (Table 33 and Table 34) and at all unscheduled visits as applicable. Vital sign measurements will be recorded with the subject in a seated position and after the subject has been resting for at least 5 minutes. All measurements will be performed singly. For any abnormal vital sign finding, further assessment should be performed as deemed necessary byAttorney Docket No.: CDPH-003 / 001WO 341287-2404 the investigator. In case of rescreening, all vital sign parameters will be rechecked as described above.

[0577] Physical examination will include assessment of heart, lung, extremities, body weight, and height. Height will only be assessed at screening. Body weight will be assessed at screening and on Day 85 (Visit 9 of Part A and Visit 8 of Part B). All the other assessments will be performed at the time-points specified in the Schedule of Assessments (Table 33 and Table 34) and at all unscheduled visits as applicable. Clinically significant changes from the initial physical findings at screening observed in subsequent visits will be considered AEs.

[0578] Resting 12-lead ECGs will be recorded after the subject has been supine and at rest for at least 5 minutes at the time-points specified in the Schedule of Assessments (Table 33 and Table 34) and at all unscheduled visits as applicable.

[0579] An adverse event (AE) is any untoward medical occurrence associated with the use of a drug in humans, whether or not considered drug related. This includes the following: y • Any clinically significant worsening of a pre-existing condition. y • Any recurrence of a pre-existing condition. y • An AE occurring from overdose of a sponsor study drug whether accidental or intentional (i.e., a dose higher than that prescribed by a health care professional for clinical reasons). y • An AE occurring from abuse of a sponsor study drug (i.e., use for non-clinical reasons). y • An AE that has been associated with the discontinuation of the use of a sponsor study y drug.

[0580] The severity of the AE will be assessed using the following categories: y Mild: Usually transient and may require only minimal treatment or therapeutic intervention. The event does not generally interfere with usual activities of daily living. y Moderate: Usually alleviated with additional specific therapeutic intervention. The event interferes with usual activities of daily living, causing discomfort but poses no significant or permanent risk of harm to the subject. y Severe: Interrupts usual activities of daily living, significantly affects clinical status, or may require intensive therapeutic intervention.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0581] All noxious and unintended responses to an investigational medicinal product (IMP; i.e., where a causal relationship between an IMP and an AE is a reasonable possibility) related to any dose should be considered adverse drug reactions.

[0582] For marketed medicinal products, a response to a drug which is noxious and unintended and which occurs at doses normally used in man for prophylaxis, diagnosis, or therapy of diseases or for modification of physiological function is to be considered an adverse drug reaction.

[0583] An unexpected adverse drug reaction is defined as an adverse reaction, the nature or severity of which is not consistent with the applicable product information.

[0584] A Serious Adverse Events (SAE) is any AE occurring at any dose that meets 1 or more of the following criteria: • Results in death • Is life threatening (see below) • Requires subject hospitalization or prolongation of an existing hospitalization (see below) • Results in a persistent or significant disability or incapacity (see below) • Results in a congenital anomaly or birth defect • Results in an important medical event (see below).

[0585] A life-threatening AE is any AE that places the subject at immediate risk of death from the event as it occurred. A life-threatening event does not include an event that might have caused death had it occurred in a more severe form but that did not create an immediate risk of death as it actually occurred.

[0586] Hospitalization or prolongation of a hospitalization is a criterion for considering an AE to be serious.

[0587] Disability is defined as a substantial disruption in a person’s ability to conduct normal life functions (i.e., the AE resulted in a significant, persistent, or permanent change, impairment, damage, or disruption in the subject’s bodily function / structure, physical activities, or quality of life). Pharmacokinetic Analysis

[0588] Blood samples for analysis of concentrations of the compound of Formula (I) (e.g., Compound A) will be collected by venipuncture at the time-points specified in the Schedule of Assessments (Table 33 and Table 34). Procedures for collection, processing, and shipping of PK blood samples will be detailed in the laboratory manual. PK samples will be assessed at a central laboratory.Attorney Docket No.: CDPH-003 / 001WO 341287-2404

[0589] Plasma concentration of the compound of Formula (I) (e.g., Compound A) will be determined using a validated analytical procedure. Specifics of the analytical method will be provided in the bioanalytical report. Sample Size And Data Analyses Determination of Sample Size

[0590] In Part A, approximately 60 subjects with CHFrEF will be included. The sample size was not determined by power calculations but by considerations to provide a reasonable number of subjects to assess the study objectives.

[0591] In Part B, approximately 600 subjects are planned to be included with approximately 300 subjects with CHFrFF and 300 subjects with CHFpEF. Subjects will be randomized 1:1, compound of Formula (I) (e.g., Compound A) to matching placebo. The randomization will be stratified on Entresto use (yes or no) and EF (≤ 40%, >40%). Regardless of EF and Entresto use, 300 subjects per treatment group will provide 99% power in the whole population using a t-test to detect an NT-proBNP reduction of 25% (i.e., a reduction of 0.288 units of log-transformed NT-proBNP) with an assumed standard deviation of 0.80 units of log-transformed BNP at Week 12 between the compound of Formula (I) (e.g., Compound A) and placebo at the one-sided 0.025 alpha level.

[0592] This sample size will also allow to have sufficient power to detect the same reduction in NTproBNP at the one-sided 0.025 alpha level in the subgroups of analysis of CHFrEF subjects and CHFpEF subjects (150 per treatment group, power=87%), subjects with Entresto use (135 per treatment group, power=84%) and subjects without Entresto use (165 per treatment group, power=90%).

[0593] There is considerable uncertainty with the uptake of Entresto use, especially among CHFpEF subjects. Because of this uncertainty, once the planned number of subjects within each stratum has been randomized, then that stratum will be closed to enrollment. For example, once 180 CHFrEF subjects on Entresto have been randomized, then the study will be closed to CHFrEF subjects on Entresto, but enrollment would continue for other subjects whose subgroup has not been closed. Use of capping measures will ensure adequate sample sizes for each subgroup to assess the study objectives. Analysis Populations

[0594] The following analysis populations will be included for this study: y Intention to Treat (ITT): The ITT population consists of all randomized subjects. Subjects will be analyzed according to their randomized treatment, regardless ofAttorney Docket No.: CDPH-003 / 001WO 341287-2404 actual treatment received, treatment compliance, or treatment duration. The ITT population will be used for sensitivity analyses. y Modified ITT (mITT): The mITT population consists of all randomized subjects who received at least one dose of the compound of Formula (I) (e.g., Compound A) or placebo. Subjects will be analyzed according to their randomized treatment, regardless of actual treatment received, treatment compliance, or treatment duration. The mITT population will be used as the primary efficacy population. y Safety: The Safety population consists of all subjects who received at least one dose of the compound of Formula (I) (e.g., Compound A) or placebo. Subjects will be analyzed according to the treatment first received. The Safety population will be used as the primary safety population. y Pharmacokinetic: The PK population consists of all subjects who received at least one dose of the compound of Formula (I) (e.g., Compound A) and have no protocol deviations that are considered to impact the PK results. y Per protocol (PP): The PP population consists of all mITT subjects with no important protocol deviations. Subjects will be analyzed according to their randomized treatment. The PP population will be used for sensitivity analyses.

[0595] In general, data will be summarized with descriptive statistics for continuous endpoints, and frequency and percentage for categorical endpoints, unless otherwise specified. Percentages by categories will be based on the number of subjects with no missing data (i.e., will add up to 100%).

[0596] Baseline will be defined as the last non-missing observation prior to first treatment intake, unless otherwise specified. In Part B, capping measures will be applied to ensure adequate sample sizes for each subgroup to assess the study objectives. Efficacy Analysis Primary Efficacy Outcome Measures

[0597] Part A: The primary endpoint is the change from baseline (Day -1) in plasma cGMP at Week 4. It will be compared between the compound of Formula (I) (e.g., Compound A) and placebo in the mITT population using an analysis of covariance (ANCOVA) model with categorical terms for treatment (2 levels), and Entresto (2 levels), and continuous terms for EF, and baseline plasma cGMP. The compound of Formula (I) (e.g., Compound A) will be analyzed as one treatment group even though the compound of Formula (I) (e.g., Compound A) consists of two weeks of treatment with 10 mg every 12 hours, followed by two weeks ofAttorney Docket No.: CDPH-003 / 001WO 341287-2404 treatment with the compound of Formula (I) (e.g., Compound A) at 25 mg every 12 hours. The least square means of the change from baseline for each treatment group, along with the 95% confidence intervals (CI), and the least square mean treatment difference and the associated 95% CI for this difference will come from the ANCOVA model.

[0598] Part B: The primary endpoint is the change from baseline in NT-proBNP at Week 12. NT-proBNP is measured at 5 time-points: Screening (Day -14 to Day -1), day 1 (pre-dose), week 2 (Day 15 ± 2, pre-dose), week 6 (Day 43 ± 3, pre-dose), week 12 (Day 85 ± 5). The baseline serum NT-proBNP is defined as the value from the Day 1 measurement. Change from baseline in serum NT-proBNP over time will be analyzed in the mITT population using a mixed model for repeated measures (MMRM) including treatment, visit, Entresto use, treatment-by-visit interaction, treatment-by-Entresto interaction, visit-by-Entresto interaction, and baseline EF as a covariate with random effects for subject, intercept, and slope. The covariance structure will be unstructured. No imputation of missing data will be performed. The least-square means and associated two-sided 95% CI for each treatment, the estimated mean treatment differences, and corresponding two-sided 95% CI coming from the MMRM will be given for all visits. The superiority of the treatment will be confirmed if the one-sided p-value for treatment-difference at Week 12 is below 0.025. The same analysis will be repeated within each one of the 8 subgroups in the mITT population.

[0599] Sensitivity analyses: y An analysis will be conducted using the MMRM from the primary analysis of the primary endpoint described above but in the PP population. y Analyses will be conducted using the MMRM from the primary analysis of the primary endpoint described above in the ITT population but different model assumptions will be investigated with a different covariance structure (i.e., AR(1), CS). y An analysis will be conducted using a non-parametric ANCOVA model using the observed baseline and Week 12 NT-proBNP values. y An analysis will be conducted using the MMRM from the primary analysis of the primary endpoint described above in the mITT population but missing serum NTproBNP values will be imputed using multiple imputation and Ruben rules. y An analysis will be conducted using the MMRM from the primary analysis of the primary endpoint described above in the PP population but missing serum NT-Attorney Docket No.: CDPH-003 / 001WO 341287-2404 proBNP values will be imputed using multiple imputation and Ruben rules. The statistical analysis plan (SAP) will provide complete details of those analyses. Secondary Efficacy Outcome Measures (Part B Only)

[0600] The change from baseline (Day 1) in BNP will be analyzed in the same way as the primary efficacy endpoint using an MMRM. Treatment difference between changes from baseline at Week 12 will be presented.

[0601] The change from baseline (Day 1) in plasma cGMP will be analyzed in the same way as the primary efficacy endpoint using an MMRM. Treatment difference between changes from baseline at Week 12 will be presented.

[0602] The change from baseline (Day 1) in plasma cGMP to NT-proBNP and cGMP to BNP ratios will be analyzed in the same way as the primary efficacy endpoint using an MMRM. Treatment difference between changes from baseline at Week 12 will be presented.

[0603] The KCCQ-23 has the clinical summary score, overall summary score, and 8 domains (physical limitations, symptom stability, symptom frequency, symptom burden, total symptom score, quality of life, self-efficacy, and social limitation).

[0604] The proportion of subjects with ≥5-point increase in the KCCQ-23-CS from baseline to Week 12 in subjects with CHF will be computed within each treatment group. Treatment effect at Week 12 will be estimated using a logistic regression with Entresto use, baseline KCCQ-23-CS score, and baseline EF as covariates. The odds ratio between treatment groups, resulting from the logistic regression, and its associated 95%CI will be provided.

[0605] In addition, descriptive statistics on the change from baseline to Week 12 for KCCQ- 23-CS score will be provided. In addition, an ANCOVA model will be used to estimate the mean treatment difference in the change from baseline at Week 12. This analysis will be repeated in the 8 subgroups of interest. Safety Analysis

[0606] All safety data will be summarized, by treatment group and dose period (Period 1, Period 2, Period 3, pooled Period 2 and Period 3, and overall) for Part A and by treatment group for Part B in the Safety Data Set.

[0607] For Part A, the primary safety analysis will be performed by reporting the exposure- adjusted incidence rates in subject-years for AE and SAE defined as the number of subjects with that particular AE or SAE within a system organ class or preferred term during the relevant period divided the sum of the at-risk times. The total at risk time is the sum of exposure time in each period in years. The adjusted incidence rates will be presented by system organ class and preferred term for Period 1, and separately for Periods 2 and 3Attorney Docket No.: CDPH-003 / 001WO 341287-2404 combined. The Hodges-Lehmann methodology will be used to estimate the within-subject point and interval treatment difference.

[0608] For both Part A and Part B, global incidence of emergent adverse events (EAE) (i.e., all events occurring, worsening or becoming serious after the first study treatment intake) will be given by system organ class and preferred term according to the Medical Dictionary of Regulatory Activities. The same description will be performed for serious EAE, severe EAE, treatment related EAE, and EAE leading to study drug withdrawal. AEs leading to death will also be summarized.

[0609] For both Part A and Part B, descriptive statistics will be performed on observed values at each visit and on each change and relative change from baseline. Box plots will be produced for some pre-selected vital signs.

[0610] For both Part A and Part B, quantitative routine 12-lead ECG results will be summarized on observed values at each visit and on each change and relative change from baseline. Box plots will be produced for some pre-selected ECGs. 12-lead ECG data interpretations and quantitative values will be listed.

[0611] For both Part A and Part B, descriptive statistics will be performed on observed values at each visit and on each change and relative change from baseline. The number and percentage of subjects having a value out of reference laboratory range (respectively out of potentially clinically significant range) will also be provided. Box plots will be produced for some pre-selected lab parameters. Pharmacokinetic Analysis

[0612] PK parameters will be calculated with plasma concentrations of the compound of Formula (I) (e.g., Compound A) in Part A using noncompartmental methods. PK parameters will not be calculated for Part B due to limited PK sampling. All plasma concentrations of the compound of Formula (I) (e.g., Compound A) will be summarized by study part, dose, visiting day, and time-points. PK parameters will be summarized for Part A by dose and visiting day. Example 4: A Phase 2, Randomized, Double-Blind, Placebo-Controlled Study to Assess the Safety and Pharmacodynamic Effects of the Compound of Formula (I) (e.g., Compound A) in Subjects with Chronic Heart Failure

[0613] The natriuretic-peptide (NP) signaling pathway plays a beneficial role in cardiovascular pathophysiology. Once stimulated, the second messenger cyclic guanosine monophosphate (cGMP) amplifies the NP signal. The importance of this signaling pathwayAttorney Docket No.: CDPH-003 / 001WO 341287-2404 in HF has been validated by multiple clinical HF studies of sacubitril / valsartan demonstrating clinical efficacy. NP signaling is decreased by PDE9-mediated hydrolysis of cGMP. Myocardial biopsies from HF patients show that PDE9 is up-regulated in both HFrEF and HFpEF, confirming that PDE9 decreases cGMP signaling generated by NP receptor activation in HF, with the potential to increase cGMP signaling via PDE-9 inhibition. PDE9 inhibition is thus an important new target for clinical studies of HF. Increase in cGMP concentrations following treatment of HF patients with a PDE9 inhibitor is a novel biomarker of potential clinical efficacy.

[0614] The safety and tolerability of PDE9 inhibition in 60 patients with HF with reduced ejection fraction (HFrEF) and the effect of the compound of Formula (I) (e.g., Compound A) compared to placebo on plasma and urinary cGMP at Week 4 are assessed.

[0615] Patients eligible for the trial were adults ≥18 years of age, with history of HF of > 6 months, NYHA class II or III, at least one of dyspnea, PND, edema, fatigue, exertional intolerance, or hospitalization or ED visit for HF requiring IV diuretic, as well as EF < 40% and NT-proBNP ≥600 pg / ml at screening or of ≥1000 pg / mL if in atrial fibrillation or flutter, on treatment with stable doses of guideline-directed HF therapy for a minimum of 4 weeks prior to screening. Eligible subjects were randomized to either the compound of Formula (I) (e.g., Compound A) or placebo in a 2:1 ratio. Participants started an oral dose of the compound of Formula (I) (e.g., Compound A) or matching placebo, for 2 weeks, followed by dose increase and treatment for an additional10 weeks, to complete a total of 12 weeks of the compound of Formula (I) (e.g., Compound A) or matching placebo.

[0616] Safety was assessed by adverse events, physical examinations, vital signs, ECGs, and laboratory safety tests. The primary pharmacodynamic endpoint is the change from baseline in plasma cGMP at Week 4 between treatment groups. The secondary endpoint is to assess the pharmacokinetics of the compound of Formula (I) (e.g., Compound A) in subjects with HFrEF. Example 5: A Phase 2, Randomized, Double-Blind, Placebo-Controlled Clinical Study to Assess the Effectiveness of the Compound of Formula (I) (e.g., Compound A) in Subjects with Chronic Heart Failure – Results of Part A Treatment-Emergent Adverse Events

[0617] Compound A was well-tolerated in a population of HFrEF patients on GDMT.

[0618] Compound A was well-tolerated in this 60-patient Phase 2 Part A trial in HFrEF patients (Table 35).Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Table 35: Toleration of Compound A on HFrEF Patients Characteristic Statistics Compound A (n=40) Placebo (n =20) R D C C C CPDE9 Inhibition with Compound A in Plasma and Urine cGMP

[0619] As shown in FIG. 3, PDE9 inhibition with Compound A led to consistent, strong increases in urine cGMP compared to placebo (Table 36 and Table 37). Table 36: Median Percent Change from Baseline in Urinary cGMP- . , . - . , . Table 37: Mean Percent Change from Baseline in Urinary cGMPWeek 2 132 212 +84%Attorney Docket No.: CDPH-003 / 001WO 341287-2404 (75, 234) (163, 274) (+14%, +198%)

[0620] A robust evaluation in the median percent change of plasma cGMP from the baseline was detected at Day 1, 15, and 29 following administration of Compound A. The evaluation in plasma cGMP occurred early after administration of Compound A and was consistent throughout the study (Table 38). The cGMP increases at all time points were statistically significant increased compared to placebo. (Table 39).

[0621] The mean percent change from the baseline for plasma cGMP was measured at Day 1, Day 15, and Day 24 following the administration of Compound A (Table 40). FIG. 4 shows that upon the administration of Compound A, the cGMP increases at all time points and remains elevated throughout the duration of Period 1 and Period 2 stages of the administration (Table 41). Table 38: Median Percent Change from Baseline in Plasma cGMP AUC, , Table 39: Median Time-matched Percent Change from Baseline in Plasma cGMP at Day 1, 15, and 29 T 1 2 36 hour 21.18 26.57 33.6 0.0029 Table 40: Mean Percent Change from Baseline in Plasma cGMP AUC(26.66) (41.69)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Week 2 13.56 26.31 12.75Table 41: Mean Percent Change (Standard Deviation) from Baseline in Plasma cGMP at Day 1, 15, 29 with 10 mg and 25 mg BID Compound A. Period 1 (10 m BID) Period 2 (25 m BID) T p 1 h 2 h 3 h 6 hEffects of Compound A and Entresto on cGMP

[0622] The evaluation in median percent change of the plasma cGMP detected at Day 1, 15, and 29 following the administration of Compound A was similar between the Entresto- treated and Entresto-naïve patients (Table 42). The Compound A-mediated plasma cGMP increases were additive to Entresto, as three-quarters of patients were taking Entresto during the trial (73% of Compound A, 75% placebo pts). As shown in FIG. 5, treatment with Compound A showed a consistently higher mean time-matched percent change from the baseline in Plasma cGMP at week 4 compared to the placebo for both Entresto-treated and Entresto-naïve patients, further demonstrating the Compound A-mediated additive effects to Entresto. Table 42: Placebo-corrected Median Time-matched Percent Change from Baseline in Plasma cGMP; Entresto-treated Patients T 1 23 hour 16.55 24.82 34.37 0.009Attorney Docket No.: CDPH-003 / 001WO 341287-2404 6 hour 18.71 16 28.53 0.02 -1.to theonNT-proBNP Levels

[0623] FIG. 6 shows that the NT-proBNP levels did not differ in the group-level analyses in this small 60 patient study (Table 43). However, a post-hoc exploratory analysis supports a potential treatment effect on NT-proBNP in patients with worse heart failure at baseline. Table 43: Post-hoc; Compound A Reduced 12-Week NTproBNP in Subgroups of HF Patients Baseline Compound A Compound A Effect Compound A Effect A [ B K 9 B K 8 B K 7 B K 6KCCQ Scores

[0624] FIG. 7 shows improvements in the mean change of multiple KCCQ domains from baseline for the Compound A group compared to placebo, with some domains approaching statistical significance (Table 44). The analysis of the NT-proBNP changes in subgroups of baseline KCCQ showed a significant treatment-related decrease in NT-proBNP in patients with lower (more severe HF) baseline KCCQ values (Table 45). As demonstrated in FIG. 8, administration of Compound A resulted in an increased percentage of patients possessing larger categorical changes relative to the placebo across 5- 10-, and 20-point increases to KCCQ Summary Scores. Table 44: Mean change in KCCQ from Baseline at Week 12Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Summary KCCQ Placebo Compound A Treatment Effect C O T K P S S F S Q S STable 45: Magnitude of Observed KCCQ Changes 5 1 i 1 i 2 iPharmacokinetics of Compound A

[0625] The plasma concentrations of Compound A were measured at Day 1 after a single dose of 10 mg BID, at week 2 after a single dose of 25 mg BID, and at week 4 after two weeks of dosing 25 mg BID. The plasma concentrations of Compound A demonstrated dose- proportional exposure between 10 mg BID and 25 mg BID and a modest degree of accumulation with 25 mg BID over the two-week dosing period (Table 46), which were consistent with Compound A exposures from other clinical studies. TaCmax(ng / mL) 96.1 (24.2) 262 (112) 317 (123)Attorney Docket No.: CDPH-003 / 001WO 341287-2404 Tmax(h) 1.5 (0.8) 1.7 (0.8) 1.9 (1.1) A1Dapplicable

Claims

Attorney Docket No.: CDPH-003 / 001WO 341287-2404 CLAIMS 1. A method of treating chronic heart failure in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound, wherein the compound has the structure of Formula (I):or a pharmaceutically R is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C3- C8)cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, each of which optionally may be substituted with one to three substituents, the substituents being independently selected from the group consisting of (C1-C4)alkyl, (C1-C4)alkoxy, halo, and (C1-C4)haloalkyl, R1is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C2-C4)alkenyl, (C2- C4)alkynyl, (C1-C4)haloalkyl, and cyclopropyl; R2is selected from the group consisting of (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1- C6)haloalkyl, heteroaryl selected from the group consisting of pyridinyl, pyridazinyl, pyrimidinyl, and pyrazinyl, and ER5, wherein the heteroaryl optionally may be substituted with one to three substituents independently selected from the group consisting of (C1- C4)alkyl and (C1-C4)haloalkyl; R3is selected from the group consisting of hydrogen, (C1-C4)alkyl, (C2-C4)alkenyl, (C2- C4)alkynyl, (C3-C6)cycloalkyl, and (C1-C4)haloalkyl; E is selected from the group consisting of —CH2—, —CH2CH2—, —CH2CH2CH2—, and — C(O)—; andR5is selected from the group consisting of (C3-C8)cycloalkyl, heterocycloalkyl, aryl, aryloxy,and heteroaryl, any of which optionally may be substituted with one to three substituents, such substituents being independently selected from the group consisting of (C1-C4)alkyl, (C2-C4)alkenyl, (C2-C4)alkynyl, (C1-C4)hydroxyalkyl, (C1-C4)haloalkyl, (C1-C4)alkoxy, (C1-C4)haloalkoxy, (C3-C8)cycloalkyl, halo, cyano, phenyl, morpholinyl, (C1- C4)alkylamino, pyrazolyl, triazolyl, and imidazolyl.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 2. A method of treating chronic heart failure in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound where the compound is 6-((3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl)-1- (tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one or a pharmaceutically acceptable salt thereof.

3. The method of claim 1 or 2, comprising administering 5 mg to 50 mg of the compound to the subject.

4. The method of claim 1 or 2, comprising administering 5 mg to 25 mg of the compound to the subject.

5. The method of claim 1 or 2, comprising administering 25 mg of the compound to the subject.

6. The method of claim 1 or 2, comprising administering 50 mg of the compound to the subject.

7. The method of any one of claims 1-6, comprising administering the compound twice daily.

8. The method of any one of claims 1-7, comprising administering the compound every 12 hours.

9. The method of any one of claims 1-8, comprising administering the compound for at least two weeks.

10. The method of any one of claims 1-9, comprising administering the compound for at least 10 weeks.

11. The method of any one of claims 1-9, comprising administering the compound for the rest of the subject’s life.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 12. The method of any one of claims 1-11, comprising administering the compound of Formula (I) orally.

13. The method of any one of claims 1-12, wherein the chronic heart failure is chronic heart failure with reduced ejection fraction.

14. The method of any one of claims 1-12, wherein the chronic heart failure is chronic heart failure with preserved ejection fraction.

15. The method of any one of claims 1-12, wherein the chronic heart failure is chronic heart failure with midrange ejection fraction.

16. The method of any one of claims 1-12, wherein the chronic heart failure is chronic heart failure with super ejection fraction.

17. The method of any one of claims 1-16, wherein prior to the administration the subject has plasma or serum NT-proBNP levels of at least 600 pg / ml.

18. The method of any one of claims 1-17, wherein the method results in a reduction in N-terminal pro b-type natriuretic peptide (NT-proBNP) levels of about 30% as measured in the plasma or serum of the subject.

19. The method of any one of claims 1-18, wherein the method results in a reduction in b- type natriuretic peptide (BNP) levels of about 30% as measured in the plasma or serum of the subject.

20. The method of any one of claims 1-19, wherein the method results in an increase in cGMP levels of about 30% as measured in the plasma or serum of the subject.

21. The method of any one of claims 1-20, wherein the method results in an increase in cGMP levels of about 30% as measured in the urine of the subject.

22. The method of any one of claims 1-21, wherein the method results in an increase in the ratio of plasma or cGMP to plasma NT-proBNP levels of about 30%.Attorney Docket No.: CDPH-003 / 001WO 341287-2404 23. The method of any one of claims 1-22, wherein the method results in an increase in the ratio of plasma cGMP to plasma BNP levels of about 30%.

24. The method of any one of claims 1-23, wherein the method results in an increase in the Kansas City Cardiomyopathy Questionnaire-23 (KCCQ-23) score of at least 5 points.

25. A method of treating chronic heart failure in a subject in need thereof, the method comprising orally administering to the subject 25 mg of 6-((3S,4S)-4-methyl-1-(pyrimidin-2- ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro-2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4- d]pyrimidin-4-one, or a pharmaceutically acceptable salt thereof, every 12 hours.

26. Use of 6-((3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro- 2H-pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of chronic heart failure in a subject in need thereof.

27. 6-((3S,4S)-4-methyl-1-(pyrimidin-2-ylmethyl)pyrrolidin-3-yl)-1-(tetrahydro-2H- pyran-4-yl)-1,5-dihydro-4H-pyrazolo[3,4-d]pyrimidin-4-one or a pharmaceutically acceptable salt thereof for use in treating chronic heart failure in a subject in need thereof.

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