Diaminocyclopentyl substituted heteroaryl derivatives, use thereof and preparation method therefor

Abstract

Provided are diaminocyclopentyl substituted heteroaryl derivatives, the use thereof and a preparation method therefor. In particular, the present disclosure relates to compounds represented by formula (I)each substituent of which is defined in the description, and a preparation method therefor. The compounds can be used as PCSK9 inhibitors to prevent and / or treat PCSK9-mediated diseases and disorders.

Description

Diaminocyclopentyl-substituted heteroaryl derivatives and their use and preparation methods Technical Field

[0001] The present invention relates to a diaminocyclopentyl-substituted heteroaryl derivative and its application and preparation method, belonging to the field of pharmaceuticals. Background Art

[0002] PCSK9, also known as "proprotein convertase subtilisin / kexin9", is a member of the secreted proprotein convertase family and plays an important role in cholesterol metabolism. PCSK9 increases circulating LDL cholesterol levels by enhancing degradation of the LDL receptor independently of its catalytic activity. Secreted PCSK9 binds to the epidermal growth factor domain A (EGFA) of the LDL receptor (LDLR) on the cell surface, and the PCSK9 / LDL receptor complex is internalized into the endosomal / lysosomal compartment. The enhanced binding affinity of PCSK9 to the LDL receptor at the acidic pH of the late endosome / lysosome reduces LDL receptor recycling and targets the LDL receptor for lysosomal degradation. Genetic association studies have demonstrated that loss-of-function mutations in PCSK9 are associated with lower plasma LDL-C levels and a reduced incidence of adverse cardiovascular events.

[0003] Another biological pathway involving the effect of PCSK9 on the LDL receptor is the onset of septic shock. Septic shock is a usually fatal complication of severe microbial infection (sepsis) that triggers an uncontrolled systemic inflammatory response and subsequent organ failure. Sepsis originates from microbial cell walls containing pathogenic lipid moieties such as lipopolysaccharide (LPS; Gram-negative bacteria). LPS is a potent ligand for mammalian innate immune receptors [Toll-like receptors (TLRs)] and therefore plays a prominent role in the septic inflammatory response (septic shock or sepsis). PCSK9 reduces LPS absorption through the liver's LDL receptors, causing free LPS to overstimulate the body's immune response to the pathogens that cause sepsis. Therefore, inhibiting PCSK9 is beneficial in maintaining liver LDL receptors to achieve systemic pathogen clearance and detoxification in response to sepsis. However, there is currently no effective treatment for sepsis or septic shock except for antibiotic therapy.

[0004] Two monoclonal antibodies (mAbs) that selectively bind to extracellular PCSK9 and prevent it from interacting with the LDL receptor, alizumab and evolocumab, have recently been approved by the FDA and can significantly lower LDL levels. However, monoclonal antibodies require intravenous injection and patient compliance is poor. Therefore, ease of dosing and application becomes a key factor in patient compliance with drug treatment. Related patent applications for PCSK9 inhibitors that have been disclosed include WO2023084449A, WO2020150473A, WO2020150474A, etc. More PCSK9 inhibitors with increased efficacy and easier administration that can be achieved by small molecule PCSK9 inhibitors are needed.

[0005] Summary of the Invention

[0006] The present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0007] in,

[0008] R 1 is selected from hydrogen or halogen;

[0009] Ring A is selected from a 3- to 7-membered cycloalkyl group or a 3- to 7-membered heterocycloalkyl group, wherein the heterocycloalkyl group contains at least one heteroatom selected from O, N or S;

[0010] Each R 2 are independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R A replace;

[0011] Alternatively, two R attached to the same atom 2 Together with the connecting atom, it forms a 3- to 6-membered cycloalkyl group or a 3- to 6-membered heterocycloalkyl group, wherein the 3- to 6-membered cycloalkyl group or the 3- to 6-membered heterocycloalkyl group is optionally substituted by one or more R A replace;

[0012] R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0013] G1 is selected from -N- or -CR 3A -;

[0014] G2 is selected from -N- or -CR 3B -;

[0015] R 3A 、R 3B and R 3 Each independently selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl, said heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from O or N, -CONR'R" or -NH-C(O)-OR'; said C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl are optionally substituted with one or more R B Substituted, the R B Selected from halogen, hydroxy, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy or 3- to 6-membered cycloalkyl;

[0016] R' and R" are each independently selected from hydrogen, C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0017] Ring B is selected from a 5- to 6-membered heterocycloalkyl group or a 5- to 6-membered heteroaryl group, wherein the 5- to 6-membered heterocycloalkyl group or the 5- to 6-membered heteroaryl group optionally contains at least one nitrogen atom and / or at least one oxo group;

[0018] Each R 4 Each independently selected from halogen, hydroxy, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl, the heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from O or N, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl are optionally substituted with one or more R C Substituted, the R C Selected from halogen, hydroxy, cyano, C1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy or 3- to 6-membered cycloalkyl;

[0019] m is selected from 0, 1, 2 or 3;

[0020] n is selected from 0, 1, 2 or 3; and

[0021] s is selected from 0, 1, 2 or 3.

[0022] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein ring A is a 3- to 7-membered cycloalkyl group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, or cycloheptyl.

[0023] In some embodiments, Ring A is cyclopentyl.

[0024] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein ring A is a 3- to 7-membered heterocycloalkyl group (e.g., a 3-membered, 4-membered, 5-membered, 6-membered, or 7-membered group), and the heterocycloalkyl group contains at least one heteroatom selected from O, N, or S.

[0025] In an alternative embodiment, the ring A is a 4- to 6-membered heterocycloalkyl group containing 1 or 2 heteroatoms selected from O, N or S.

[0026] In an alternative embodiment, the ring A is a 5- to 6-membered heterocycloalkyl group containing 1 or 2 heteroatoms selected from O, N or S.

[0027] In an alternative embodiment, the ring A is a 5- to 6-membered heterocycloalkyl group containing 1 or 2 O atoms.

[0028] In an alternative embodiment, the ring A is a 5- to 6-membered heterocycloalkyl group containing 1 O atom.

[0029] In an alternative embodiment, the ring A is a 5- to 6-membered heterocycloalkyl group containing 2 O atoms.

[0030] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (I-1) or a pharmaceutically acceptable salt thereof,

[0031] Among them, ring B, R 1 、R 2 , G1, G2, R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (I-2) or a pharmaceutically acceptable salt thereof,

[0033] Among them, ring B, R 1 、R 2 , G1, G2, R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0034] In an optional embodiment, the compound or pharmaceutically acceptable salt thereof provided by the present disclosure (I) is a compound or pharmaceutically acceptable salt thereof represented by formula (I-3),

[0035] Among them, ring B, R 1 , G1, G2, R 3 、R 4 , n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0036] In an optional embodiment, the compound or pharmaceutically acceptable salt thereof provided by the present disclosure (I) is a compound or pharmaceutically acceptable salt thereof represented by formula (I-4).

[0037] Among them, ring B, R 1 , G1, G2, R 3 、R 4 , n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0038] In an optional embodiment, the compound or pharmaceutically acceptable salt thereof provided by the present disclosure (I) is a compound or pharmaceutically acceptable salt thereof represented by formula (I-5),

[0039] Among them, ring B, R 1 , G1, G2, R 3 、R 4 , n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0040] In an optional embodiment, the compound or pharmaceutically acceptable salt thereof provided by the present disclosure (I) is a compound or pharmaceutically acceptable salt thereof represented by formula (I-6),

[0041] Among them, ring B, R 1 , G1, G2, R 3 、R 4 , n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0042] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), or pharmaceutically acceptable salts thereof, wherein G1 is -N-; and G2 is -CR 3B -.

[0043] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein G1 is -N-; and G2 is -N-.

[0044] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein G1 is -CR 3A -; and G2 is -CR 3B -.

[0045] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is selected from

[0046] In an optional embodiment, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is

[0047] In an optional embodiment, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is

[0048] In an optional embodiment, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is

[0049] In an optional embodiment, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is

[0050] In an optional embodiment, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein ring B is

[0051] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein each R 4 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano.

[0052] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6), or pharmaceutically acceptable salts thereof, wherein each R 4 Each is independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl.

[0053] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein R 3A 、R 3B and R 3 are independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano.

[0054] In some embodiments, the present disclosure provides compounds represented by formula (I), (I-1), (I-2), (I-3), (I-4), (I-5), (I-6) or pharmaceutically acceptable salts thereof, wherein R 3A 、R 3B and R 3 Each is independently selected from hydrogen, fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl.

[0055] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-A-1) or a pharmaceutically acceptable salt thereof,

[0056] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0057] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-A-2) or a pharmaceutically acceptable salt thereof,

[0058] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0059] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-A-3) or a pharmaceutically acceptable salt thereof,

[0060] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0061] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-A-4) or a pharmaceutically acceptable salt thereof,

[0062] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0063] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-A-5) or a pharmaceutically acceptable salt thereof,

[0064] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0065] In some embodiments, the present disclosure provides compounds represented by formula (II-A-1), (II-A-2), (II-A-3), (II-A-4), (II-A-5) or pharmaceutically acceptable salts thereof, wherein:

[0066] R 1 is hydrogen;

[0067] Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R A Substituted, the R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0068] Each R 3 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0069] Each R 4Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0070] m, n and s are each independently selected from 0, 1, 2 or 3.

[0071] In some embodiments, the present disclosure provides compounds represented by formula (II-A-1), (II-A-2), (II-A-3), (II-A-4), (II-A-5) or pharmaceutically acceptable salts thereof, wherein:

[0072] R 1 is hydrogen;

[0073] Each R 3 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl or trifluoromethoxy;

[0074] Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0075] m is 0;

[0076] n and s are each independently selected from 0, 1 or 2.

[0077] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-B-1) or a pharmaceutically acceptable salt thereof,

[0078] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0079] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-B-2) or a pharmaceutically acceptable salt thereof,

[0080] Among them, R 1 、R 2、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0081] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-B-3) or a pharmaceutically acceptable salt thereof,

[0082] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0083] In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-B-4) or a pharmaceutically acceptable salt thereof,

[0084] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound of formula (I) or a pharmaceutically acceptable salt thereof. In some embodiments, the compound of formula (I) or a pharmaceutically acceptable salt thereof provided by the present disclosure is a compound of formula (II-B-5) or a pharmaceutically acceptable salt thereof,

[0085] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0086] In an optional embodiment, the present disclosure provides compounds represented by formula (II-B-1), (II-B-2), (II-B-3), (II-B-4), (II-B-5) or pharmaceutically acceptable salts thereof, wherein,

[0087] R 1 is hydrogen;

[0088] R 2 Selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R A Substituted, the R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0089] Each R 3 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0090] Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0091] m, n and s are each independently selected from 0, 1, 2 or 3.

[0092] In an optional embodiment, the present disclosure provides compounds represented by formula (II-B-1), (II-B-2), (II-B-3), (II-B-4), (II-B-5) or pharmaceutically acceptable salts thereof, wherein,

[0093] R 1 is hydrogen;

[0094] Two R's attached to the same carbon atom 2 Together with the connecting carbon atom, it forms a 3 to 6-membered cycloalkyl group, which is optionally substituted by one or more R A Substituted, the R ASelected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0095] Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0096] Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0097] m, n and s are each independently selected from 0, 1, 2 or 3.

[0098] In an optional embodiment, the present disclosure provides compounds represented by formula (II-B-1), (II-B-2), (II-B-3), (II-B-4), (II-B-5) or pharmaceutically acceptable salts thereof, wherein,

[0099] R 1 is hydrogen;

[0100] Each R 2 are each independently selected from fluoro or methyl;

[0101] Each R 3 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl or trifluoromethoxy;

[0102] Each R 4each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0103] m, n and s are each independently selected from 0, 1, 2 or 3.

[0104] In an optional embodiment, the present disclosure provides compounds represented by formula (II-B-1), (II-B-2), (II-B-3), (II-B-4), (II-B-5) or pharmaceutically acceptable salts thereof, wherein,

[0105] R 1 is hydrogen;

[0106] Two R's attached to the same carbon atom 2 Together with the connecting carbon atom, it forms a cyclopropyl group, which is optionally substituted by one or more R A Substituted, the R A selected from halogen;

[0107] Each R 3 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl or trifluoromethoxy;

[0108] Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0109] m, n and s are each independently selected from 0, 1, 2 or 3.

[0110] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II-C-1) or a pharmaceutically acceptable salt thereof,

[0111] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0112] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II-C-2) or a pharmaceutically acceptable salt thereof,

[0113] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0114] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II-C-3) or a pharmaceutically acceptable salt thereof,

[0115] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0116] In some embodiments, the present disclosure provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, which is a compound of formula (II-C-4) or a pharmaceutically acceptable salt thereof,

[0117] Among them, R 1 、R 2 、R 3 、R 4 , m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0118] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-1), (II-C-2) or pharmaceutically acceptable salts thereof, wherein,

[0119] R 1 For self-hydrogen;

[0120] Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R A replace;

[0121] R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0122] Each R 3 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0123] Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0124] m, n and s are each independently selected from 0, 1, 2 or 3.

[0125] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-1), (II-C-2) or pharmaceutically acceptable salts thereof, wherein,

[0126] R 1 is hydrogen;

[0127] Each R 3 Each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy;

[0128] Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0129] m is 0;

[0130] n and s are each independently selected from 0, 1, 2 or 3.

[0131] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-3), (II-C-4) or pharmaceutically acceptable salts thereof, wherein,

[0132] R 1 is hydrogen;

[0133] Each R 2 are each independently selected from halogen, C 1-6Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R A Substituted, the R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0134] Each R 3 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0135] Each R 4 Each independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0136] m, n and s are each independently selected from 0, 1, 2 or 3.

[0137] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-3), (II-C-4) or pharmaceutically acceptable salts thereof,

[0138] R 1 is hydrogen;

[0139] Two R's attached to the same carbon atom 2 Together with the connecting carbon atom, it forms a 3 to 6-membered cycloalkyl group, which is optionally substituted by one or more R A Substituted, the R A Selected from halogen, hydroxy, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 hydroxyalkyl;

[0140] Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R B Substituted, the R B is selected from halogen, hydroxy or cyano;

[0141] Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6 membered cycloalkyl is optionally substituted with one or more R C Substituted, the R C is selected from halogen, hydroxy or cyano;

[0142] m, n and s are each independently selected from 0, 1, 2 or 3.

[0143] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-3), (II-C-4) or pharmaceutically acceptable salts thereof,

[0144] R 1 is hydrogen;

[0145] Each R 2 are each independently selected from fluoro or methyl;

[0146] Each R 3each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl or trifluoromethoxy;

[0147] Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0148] m, n and s are each independently selected from 0, 1, 2 or 3.

[0149] In an optional embodiment, the present disclosure provides compounds represented by formula (II-C-3), (II-C-4) or pharmaceutically acceptable salts thereof,

[0150] R 1 is hydrogen;

[0151] Two R's attached to the same carbon atom 2 Together with the connecting carbon atom, it forms a cyclopropyl group, which is optionally substituted by one or more R A Substituted, the R A is a halogen;

[0152] Each R 3 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl or trifluoromethoxy;

[0153] Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl;

[0154] m, n and s are each independently selected from 0, 1, 2 or 3.

[0155] The present disclosure further provides the following compounds or pharmaceutically acceptable salts thereof,

[0156] Group A

[0157] Group B

[0158] Group C

[0159] Group D

[0160] Another aspect of the present disclosure provides an isotopic substitution of the aforementioned compound or a pharmaceutically acceptable salt thereof, wherein the isotopic substitution is a deuterated substance.

[0161] The present disclosure further provides a pharmaceutical composition comprising the aforementioned compound or a pharmaceutically acceptable salt thereof, an isotope substitute, and at least one pharmaceutically acceptable excipient.

[0162] In an optional embodiment, the pharmaceutical composition contains an effective amount of the aforementioned compound or a pharmaceutically acceptable salt thereof, or an isotope substitute.

[0163] Another aspect of the present disclosure provides the aforementioned compound or a pharmaceutically acceptable salt or isotope-substituted product thereof for use as a medicine.

[0164] Another aspect of the present disclosure provides use of the aforementioned compound or a pharmaceutically acceptable salt, isotope-substituted product, or composition thereof in the preparation of a drug for a PCSK9 inhibitor.

[0165] Another aspect of the present disclosure provides use of the aforementioned compound or a pharmaceutically acceptable salt, isotope substituted product, or composition thereof in the preparation of a medicament for preventing and / or treating diseases and disorders.

[0166] The present disclosure further provides a method for preventing and / or treating diseases and disorders, comprising administering the aforementioned compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof.

[0167] The present disclosure further provides the aforementioned compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof, for use in preventing and / or treating diseases and disorders.

[0168] The diseases and conditions described in the present disclosure are selected from dyslipidemia, dyslipoproteinemia, hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, hyperlipoproteinemia, xanthomas, hypoalphalipoproteinemia, sitosterolemia, atherosclerosis, arteriosclerosis, metabolic syndrome, coronary heart disease, peripheral vascular disease, congestive heart failure, stroke, vascular dementia, coronary artery disease, chronic kidney disease, retinopathy, inflammation, diabetic complications, or thrombosis.

[0169] In certain embodiments, the diseases and conditions are selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, or coronary artery disease.

[0170] In certain embodiments, the disease is hypercholesterolemia, such as familial hypercholesterolemia or autosomal dominant hypercholesterolemia.

[0171] In certain embodiments, the disease is hyperlipidemia.

[0172] In certain embodiments, the disease is coronary artery disease.

[0173] On the other hand, the present disclosure provides a use of the aforementioned compound or its pharmaceutically acceptable salt, isotope substitution, or composition in the preparation of a drug for lowering lipoprotein (a) (LP(a)), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), triglyceride-rich lipoprotein (TRL), and triglycerides.

[0174] Another aspect of the present disclosure provides use of the aforementioned compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof in the preparation of a medicament for preventing and / or treating diseases and disorders mediated by PCSK9.

[0175] The present disclosure further provides a method for preventing and / or treating diseases and disorders mediated by PCSK9, comprising administering the aforementioned compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof.

[0176] The present disclosure further provides the aforementioned compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof, for use in preventing and / or treating PCSK9-mediated diseases and disorders.

[0177] The PCSK9-mediated diseases and conditions described in the present disclosure are selected from dyslipidemia, dyslipoproteinemia, hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, hyperlipoproteinemia, xanthomas, hypoalphalipoproteinemia, sitosterolemia, atherosclerosis, arteriosclerosis, metabolic syndrome, coronary heart disease, peripheral vascular disease, congestive heart failure, stroke, vascular dementia, coronary artery disease, chronic kidney disease, retinopathy, inflammation, diabetic complications or thrombosis.

[0178] In certain embodiments, the PCSK9-mediated diseases and conditions are selected from hypercholesterolemia, hyperlipidemia, hyperlipoproteinemia, hypertriglyceridemia, dyslipidemia, dyslipoproteinemia, atherosclerosis, hepatic steatosis, metabolic syndrome, or coronary artery disease.

[0179] In certain embodiments, the disorder is hypercholesterolemia, such as familial hypercholesterolemia or autosomal dominant hypercholesterolemia.

[0180] In certain embodiments, the disease is hyperlipidemia.

[0181] In certain embodiments, the disease is coronary artery disease.

[0182] On the other hand, the present disclosure provides a method for reducing lipoprotein (a) (LP(a)), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), triglyceride-rich lipoprotein (TRL), and triglycerides, which comprises administering the aforementioned compound or its pharmaceutically acceptable salt, isotope substitution, or composition.

[0183] On the other hand, the present disclosure provides a compound or a pharmaceutically acceptable salt, isotope substitute, or composition thereof for reducing lipoprotein (a) (LP(a)), low-density lipoprotein (LDL), very low-density lipoprotein (VLDL), triglyceride-rich lipoprotein (TRL), and triglycerides.

[0184] Subjects with gain-of-function mutations in the PCSK9 gene also benefit from treatment with the disclosed compounds and compositions to counteract the mutation by inhibiting PCSK9.

[0185] In some embodiments, the methods of preventing and / or treating diseases and conditions or methods of preventing and / or treating PCSK9-mediated diseases and conditions provided by the present disclosure are administered to humans.

[0186] In an alternative embodiment, the subject of administration is a patient.

[0187] In an alternative embodiment, the subject of administration is a healthy subject.

[0188] In an optional embodiment, the applicable amount of the aforementioned compound or its pharmaceutically acceptable salt, isotope substitute, or composition is an effective amount.

[0189] In some embodiments, the unit dose of the pharmaceutical composition is 0.001 mg-1000 mg.

[0190] In certain embodiments, the pharmaceutical composition contains 0.01-99.99% of the aforementioned compound or its pharmaceutically acceptable salt or its isotopic substitution, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition contains 0.1-99.9% of the aforementioned compound or its pharmaceutically acceptable salt or its isotopic substitution. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of the aforementioned compound or its pharmaceutically acceptable salt or its isotopic substitution. In certain embodiments, the pharmaceutical composition contains 1%-99% of the aforementioned compound or its pharmaceutically acceptable salt or its isotopic substitution. In certain embodiments, the pharmaceutical composition contains 2%-98% of the aforementioned compound or its pharmaceutically acceptable salt or its isotopic substitution.

[0191] In certain embodiments, the pharmaceutical composition comprises 0.01% to 99.99% of a pharmaceutically acceptable excipient, based on the total weight of the composition. In certain embodiments, the pharmaceutical composition comprises 0.1% to 99.9% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 0.5% to 99.5% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 1% to 99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition comprises 2% to 98% of a pharmaceutically acceptable excipient.

[0192] The pharmaceutically acceptable salts of the compounds described in the present disclosure may be selected from inorganic salts or organic salts.

[0193] Another aspect of the present disclosure provides a method for preparing the aforementioned compound or a pharmaceutically acceptable salt thereof, comprising the steps of reacting a compound represented by formula (Ia) with a compound represented by formula (Ib) under alkaline conditions, optionally in the presence of a catalyst,

[0194] LG is selected from halogen or -S(O)2R 7 , the R 7 Selected from C 1-6 Alkyl or halogenated C 1-6 alkyl; and

[0195] R 1 、R 2 , G1, G2, R 3 、R 4 , Ring A, Ring B, m, n and s are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0196] In some embodiments, the LG is selected from halogen, such as fluorine or chlorine.

[0197] In some embodiments, the LG is selected from -S(O)2R 7 , the R 7 Optional methyl or ethyl etc.

[0198] In some embodiments, the basic environment is provided by an inorganic base or an organic base, such as sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, pyridine, triethylamine, N,N-diisopropylethylamine, and the like.

[0199] In some embodiments, a catalyst suitable for the Buchwald-Hartwig reaction is suitable for the reaction, and specifically dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) and the like can be selected.

[0200] The present disclosure further provides a compound represented by formula (Ib),

[0201] in,

[0202] LG is selected from halogen or -S(O)2R 7 , R 7 Selected from C 1-6 Alkyl or halogenated C 1-6 Alkyl; R 1 、R 2 and m are as defined in the compound represented by formula (I) or a pharmaceutically acceptable salt thereof.

[0203] The compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All of these isomers and their mixtures are included within the scope of the present disclosure. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

[0204] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0205] In the chemical structures of the compounds disclosed herein, the bond Indicates that the configuration is not specified, that is, if chiral isomers exist in the chemical structure, the bond Can be or or include both and Two configurations. In the chemical structure of the compound disclosed in the present invention, the bond No configuration is specified, i.e. the bond The configuration can be E-type or Z-type, or include both E and Z configurations.

[0206] The compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerizations.

[0207] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where 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 the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C. 14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0208] Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have at least 1000 times the abundance of deuterium greater than the natural abundance of deuterium (which is 0.015%) (i.e., at least 10% deuterium incorporation). In the example, the compound has a natural abundance greater than deuterium that can be at least 1000 times the abundance of deuterium, at least 2000 times the abundance of deuterium, at least 3000 times the abundance of deuterium, at least 4000 times the abundance of deuterium, at least 5000 times the abundance of deuterium, at least 6000 times the abundance of deuterium or more abundant deuterium. The disclosure also includes various deuterated forms of formula (I) compounds. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of formula (I) compounds with reference to relevant literature. Commercially available deuterated starting materials may be used in the preparation of deuterated forms of the compounds of formula (I), or they may be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0209] "Optionally" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "optionally substituted C 1-6 The term "alkyl" means that halogen or cyano may but need not be present, and the description includes both the case where the alkyl is substituted by halogen or cyano and the case where the alkyl is not substituted by halogen and cyano.

[0210] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0211] "Pharmaceutically acceptable excipients" or "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the U.S. Food and Drug Administration (FDA) for use by humans or domestic animals.

[0212] As used herein, an "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition to be treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

[0213] "Alkyl" refers to a saturated aliphatic hydrocarbon group. Non-limiting examples of alkyl groups containing 1 to 6 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, and various branched chain isomers thereof.

[0214] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like.

[0215] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, excluding the ring portion of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 4 to 10 ring atoms. Non-limiting examples of "heterocycloalkyl" include:

[0216] The heterocycloalkyl ring may be fused to an aryl or heteroaryl ring, wherein the ring attached to the parent structure is a heterocycloalkyl, non-limiting examples of which include:

[0217] wait.

[0218] The term "alkoxy" refers to -O-(alkyl), wherein alkyl is as defined above. Non-limiting examples of alkoxy include methoxy, ethoxy, propoxy, butoxy.

[0219] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring attached to the parent structure is the aryl ring, non-limiting examples of which include:

[0220] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 6 to 12-membered, more preferably 5-membered or 6-membered. For example, non-limiting examples include: imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl, triazolyl, indazolyl, benzimidazolyl, wait.

[0221] The heteroaryl ring may be fused to an aryl, heterocycloalkyl or cycloalkyl ring, wherein the ring attached to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0222] The term "hydroxy" refers to an -OH group.

[0223] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0224] The term "cyano" refers to -CN.

[0225] The term "amino" refers to -NH2.

[0226] The term "nitro" refers to -NO2.

[0227] The term "oxo" refers to a =0 substituent.

[0228] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are independently replaced by a corresponding number of substituents. When the substituent is keto or oxo (i.e., =O), then two (2) hydrogen atoms on the atom are replaced. DETAILED DESCRIPTION

[0229] The preparation of the compounds and pharmaceutically acceptable salts disclosed herein is further described below with reference to examples, but these examples are not intended to limit the scope of the disclosure.

[0230] Experimental methods in the examples of this disclosure that do not specify specific conditions are generally based on conventional conditions or the conditions recommended by the raw material or product manufacturers. Reagents without specific sources are conventional reagents purchased from the market.

[0231] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (LCMS). -6The unit of measurement is ppm. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The spatial configuration of the optical isomers (isomers) of the compounds can be further confirmed by measuring single crystal parameters.

[0232] HPLC determinations were performed using Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200LC high pressure liquid chromatograph (ACQUITY UPLC BEH C18 1.7UM 2.1*50MM column, Ultimate XB-C18 3.0*150mm column, or Xtimate C18 2.1*30mm column).

[0233] MS was determined using a Waters SQD2 mass spectrometer in positive / negative ion mode with a mass scan range of 100-1200.

[0234] The thin layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) is 0.15mm~0.2mm, and the specification used for thin layer chromatography separation and purification products is 0.4mm~0.5mm.

[0235] The flash column purification system used was Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage).

[0236] Forward column chromatography generally uses Yantai Huanghai silica gel 100-200 mesh, 200-300 mesh or 300-400 mesh silica gel as a carrier, or uses Changzhou Santai pre-packed ultra-pure normal phase silica gel column (40-63μm, 12g, 25g, 40g, 80g or other specifications).

[0237] Reversed phase column chromatography generally uses Changzhou Santai pre-packed ultra-pure C18 silica gel column (20-45μm, 40g, 80g, 120g, 220g or other specifications).

[0238] The high-pressure column purification system uses Waters AutoP, in conjunction with Waters XBridge BEH C18 OBD Prep Column. 5μm, 19mm X 150mm or Atlantis T3OBD Prep Column, 5μm, 19mm X 150mm.

[0239] The chiral preparative column used was DAICEL CHIRALPAK IC (250 mm*30 mm, 10 μm), DAICEL CHIRALCEL OJ (250 mm*30 mm, 10 μm) or DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm).

[0240] The known starting materials in the present disclosure can be synthesized by methods known in the art, or can be purchased from Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Darui Chemicals and other companies.

[0241] Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

[0242] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1 L.

[0243] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0244] The pressurized hydrogenation reaction uses a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogen generator or an HC2-SS hydrogenator.

[0245] The hydrogenation reaction is usually carried out by evacuating the chamber and filling it with hydrogen, and the operation is repeated three times.

[0246] A CEM Discover-S 908860 microwave reactor was used for the microwave reaction.

[0247] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0248] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0249] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compound, and the developing solvent system for thin layer chromatography included: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system, C: petroleum ether / ethyl acetate system, D: petroleum ether / ethyl acetate / methanol, and E: petroleum ether / tetrahydrofuran system. The volume ratio of the solvent was adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could also be added for adjustment.

[0250] Example 1

[0251] 1-(6-{[(1S,3S)-3-(6,7-dihydro-5H-cyclopenta[1,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-1,2-dihydropyridin-2-one

[0252] Step 1:

[0253] Pyridin-2(1H)-one 1b (1.51 g, 15.9 mmol), 2-chloro-5-pyridineboronic acid 1a (5 g, 31.8 mmol), copper acetate (5.77 g, 31.8 mmol), pyridine (2.51 g, 331.8 mol), and molecular sieves (6 g) were added to a mixed solution (DMF / DCM = 1:6) (210 mL), purged with air for one minute, and reacted at room temperature for 16 hours. The reaction mixture was filtered through celite, and the filtrate was added with 100 mL of water and 50 mL of ethyl acetate. The layers were separated, and the aqueous phase was extracted with ethyl acetate (50 mL x 2). The organic phases were combined and concentrated under reduced pressure. The residue was separated by normal phase column chromatography to obtain compound 1c (1.03 g, 31.4% yield).

[0254] MS m / z(ESI):207.1[M+H] + .

[0255] Step 2:

[0256] Compound 1c (1.03 g, 4.99 mmol), (1S,3S)-tert-butyl 3-aminocyclopentylcarbamate 1d (1.2 g, 5.98 mmol), cesium carbonate (4.87 g, 14.96 mmol), and methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) palladium (II) (0.42 g, 0.5 mmol) were added to 1,4-dioxane (10 mL) and microwave-treated at 130°C for 4 hours. The reaction solution was separated by normal phase column chromatography to obtain compound 1e (440 mg, yield: 23.8%).

[0257] MS m / z(ESI):371.3[M+H] + .

[0258] Step 3:

[0259] Compound 1e (440 mg, 1.2 mmol) was added to dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added. The mixture was reacted at room temperature for 1 hour and concentrated under reduced pressure to obtain compound 1f (800 mg, yield: 135.1%).

[0260] MS m / z(ESI):271.2[M+H] + .

[0261] Step 4:

[0262] Compound 1f (100 mg, 0.2 mmol), 2-chloro-6,7-dihydro-5H-cyclopenta[1,2-d]pyrimidine 1 g (34.1 mg, 0.22 mmol), and potassium carbonate (83.3 mg, 0.6 mmol) were added to DMSO (1 mL) in sequence and the mixture was heated to 110°C for 6 hours. Compound 1 was then directly isolated by reverse-phase column chromatography to afford compound 1 (4 mg, 5.1% yield).

[0263] MS m / z(ESI):389.3[M+H] + .

[0264] 1 H NMR(400MHz,MeOD)δ8.03(s,1H),7.98(d,J=2.6Hz,1H),7.67–7.51(m,3H),6.73(d,J=9.1Hz,1H),6.62(dd,J=9.1,1.2Hz,1H),6.47(td,J=6.8, 1.4Hz,1H),4.45(s,1H),4.35(t,J=6.4Hz,1H),2.88(dt,J=12.9,7.3Hz ,4H),2.35–2.26(m,2H),2.19–2.03(m,4H),1.68(qd,J=8.9,6.1Hz,2H).

[0265] Example 2

[0266] 1-(6-{[(1S,3S)-3-[(7,7-difluoro-6,7-dihydro-5H-cyclopenta[1,2-d]pyrimidin-2-yl)amino]cyclopentyl]amino}pyridin-3-yl)-1,2-dihydropyridin-2-one

[0267] Step 1:

[0268] Under a hydrogen atmosphere, compound 2a (200 mg, 0.85 mmol, synthesis method referenced in patent application WO201528848) was dissolved in methanol (3 mL), followed by the addition of 10% Pd / C (20 mg). The reaction was allowed to proceed at room temperature for 3 hours. The reaction solution was filtered and concentrated to obtain a crude product, which was then purified by normal-phase column chromatography to afford compound 2b (66 mg, 38% yield).

[0269] MS m / z(ESI):203.1[M+H] + .

[0270] Step 2:

[0271] Compound 2b (66 mg, 0.33 mmol) was dissolved in dichloromethane (5 mL), followed by the addition of 3-chloroperoxybenzoic acid (168 mg, 0.98 mmol). The reaction was allowed to react at room temperature for 2 hours. After completion of the reaction, the crude product was directly spin-dried to obtain a crude product, which was then purified by normal phase column chromatography to obtain compound 2c (45 mg, 59% yield).

[0272] MS m / z(ESI):235.1[M+H] + .

[0273] Step 3:

[0274] Compound 2c (15 mg, 0.06 mmol) and compound 1f (34.6 mg, 0.13 mmol) were dissolved in tert-butanol (2 mL), followed by the addition of N,N-diisopropylethylamine (24 μL, 0.12 mmol). The reaction was placed in a microwave oven at 110°C for 4 hours. After completion of the reaction, the crude product was directly dried to obtain a crude product, which was then purified by reverse phase column chromatography to obtain compound 2 (1.5 mg, yield: 5%).

[0275] MS m / z(ESI):425.4[M+H]+.

[0276] 1 H NMR (400MHz, MeOD) δ8.37(s,1H),7.94(d,J=2.6Hz,1H),7.66–7.55(m,2H),7.44(dd,J=9.0,2.7Hz,1H),6.66–6.57(m,2H),6.47(td,J=6.8,1. 3Hz,1H),4.53–4.44(m,1H),4.40–4.31(m,1H),2.91–2.82(m,2H),2.61 –2.46(m,2H),2.36–2.20(m,2H),2.10–1.95(m,2H),1.70–1.54(m,2H).

[0277] Example 3

[0278] 1-(6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-1,2-dihydropyridin-2-one

[0279] Step 1:

[0280] 2,4-Dichloro-5-hydroxypyrimidine 3a (520 mg, 3.2 mmol), 2-bromoethanol (1.18 g, 9.5 mmol), and potassium carbonate (2.18 g, 15.8 mmol) were added sequentially to DMF (5 mL). The reaction mixture was microwaved at 100°C for 1 hour. The reaction mixture was poured into water (15 mL) and extracted with ethyl acetate (20 mL x 2). The organic phases were combined and concentrated under reduced pressure to afford the crude product, which was then separated by normal phase column chromatography to afford compound 3b (120 mg, 22.1% yield).

[0281] MS m / z(ESI):173.1[M+H] + .

[0282] Step 2:

[0283] 3b (100 mg, 0.37 mmol), compound 1f (53 mg, 0.24 mmol), cesium carbonate (361.6 mg, 1.11 mmol), and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridinyl)palladium(II) (29.3 mg, 0.04 mmol) were added to 1,4-dioxane (1 mL). The reaction mixture was microwaved at 100°C for 2 hours. The mixture was filtered and concentrated to obtain a crude product, which was then separated by reverse-phase column chromatography to afford compound 3 (5 mg, 3.33% yield).

[0284] MS m / z(ESI):407.3[M+H] + .

[0285] 1H NMR(400MHz,MeOD)δ7.93(d,J=2.7Hz,1H),7.78(s,1H),7.60(ddt,J=11.1,6.7 ,2.0Hz,2H),7.44(dd,J=9.0,2.7Hz,1H),6.61(ddd,J=9.0,2.9,1.7Hz,2H),6.4 6(td,J=6.8,1.3Hz,1H),4.49–4.43(m,2H),4.30(dp,J=13.4,6.5Hz,2H),4.21– 4.15(m,2H),2.31–2.15(m,2H),1.95(qd,J=6.5,2.6Hz,2H),1.63–1.51(m,2H).

[0286] Example 4

[0287] 2-(5-Fluoro-6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-3H,2H-1,2-pyridazin-3-one

[0288] Step 1:

[0289] Compound 3b (2 g, 11.6 mmol), tert-butyl (1S,3S)-3-aminocyclopentylcarbamate (2.8 g, 13.92 mmol), cesium carbonate (11.34 g, 34.8 mmol), and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (550 mg, 0.70 mmol) were weighed into a reaction flask. Dioxane (20 mL) was added and the mixture was reacted at 110°C overnight under nitrogen. LCMS analysis confirmed the formation of the product. The reaction solution was concentrated and subjected to silica gel column chromatography to obtain compound 4a (2.5 g, yield: 64.1%).

[0290] MS m / z(ESI):337.2[M+H] + .

[0291] Step 2:

[0292] Compound 4a (2.5 g, 7.4320 mmol) was dissolved in DCM (25 mL), and TFA (25 mL) was added. The mixture was reacted at room temperature overnight. The product was detected by LCMS. The reaction solution was directly evaporated to dryness to obtain compound 4c (5.5 g) as a brown oil, which was directly used for the next step.

[0293] MS m / z(ESI):237.2[M+H] + .

[0294] Step 3:

[0295] Pyridazin-3(2H)-one (548 mg, 5.70 mmol) was dissolved in 10 mL of a 1:6 DMF / DCM solution. Compound 4b (500 mg, 2.85 mmol), copper acetate (1.0 g, 5.70 mmol), pyridine (0.46 mL, 5.70 mmol), and molecular sieves were then added sequentially. The mixture was stirred at room temperature for 16 hours. After completion of the reaction, the reaction mixture was filtered through celite. 10 mL of water and 30 mL of ethyl acetate were added to the filtrate, and the mixture was stirred at room temperature for 15 minutes. Ethyl acetate was then added and extracted several times (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (PE:EA = 5:1 to 1:1) to afford compound 4d (510 mg, 79.3% yield).

[0296] MS m / z(ESI):226.1[M+H] + .

[0297] Step 4:

[0298] Compound 4c (990 mg, 1.33 mmol), compound 4d (300 mg, 1.3 mmol), cesium carbonate (6.5 g, 19.9 mmol), and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (63.3 mg, 79.78 μmol) were dissolved in dioxane (30 mL) and reacted at 110°C for 4 h under nitrogen protection. The product was detected by LCMS. The reaction solution was cooled to room temperature, and water and ethyl acetate were added for separation. The aqueous phase was extracted with ethyl acetate, and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography to obtain compound 4 (375 mg, yield: 66.44%).

[0299] MS m / z(ESI):426.2[M+H] + .

[0300] 1H NMR(400MHz,DMSO-d6)δ8.08-7.99(m,2H),7.83(s,1H),7.62(m,1H),7.48(m,1H),7.05(m,1H),6.91(m,1H),6.76(m,1H), 4.49(m,1H),4.43-4.35(m,2H),4.23(m,1H),4.17-4.10(m,2H),2.18-2.01(m,2H),1.96-1.83(m,2H),1.62-1.40(m,2H).

[0301] Example 5

[0302] 2-(6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-3H,2H-1,2-pyridazin-3-one

[0303] Step 1:

[0304] Pyridazin-3(2H)-one (1 g, 10.4 mmol) was dissolved in 120 mL of a mixed solution (DMF / DCM = 1:6). Compound 5a (2.4 g, 15.3 mmol), copper acetate (3.8 g, 20.8 mmol), pyridine (1.6 g, 20.8 mmol), and 4 g of molecular sieves were then added sequentially. The reaction mixture was stirred at room temperature for 16 h. The reaction mixture was then filtered through celite. The filtrate was added with 10 mL of water and 20 mL of ethyl acetate, and stirred at room temperature for 15 min. The mixture was extracted repeatedly with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography using an eluent system (PE:EA = 5:1 to 1:1) to afford the title compound 5b (500 mg, 23.4% yield).

[0305] MS m / z(ESI):208.3[M+H] +

[0306] Step 2:

[0307] Compound 4c (1.07 g, 1.45 mmol), compound 5b (300 mg, 1.45 mmol), cesium carbonate (7.06 g, 21.67 mmol), and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (68.8 mg, 86.69 μmol) were dissolved in dioxane (30 mL) and reacted at 110°C for 4 hours under nitrogen. LCMS analysis confirmed the formation of the target compound. The reaction solution was cooled to room temperature and filtered. The filtrate was evaporated to dryness, and water and ethyl acetate were added. The layers were separated, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography to obtain compound 5 (300 mg, yield: 48.9%).

[0308] MS m / z(ESI):408.1[M+H] + .

[0309] 1 H NMR(400MHz,DMSO-d6)δ8.10(m,1H),8.01(m,1H),7.83(s,1H),7.54-7.42(m,2H),7.02(m,1H),6.90(m,1H),6.76(m,1H), 6.52(m,1H),4.44-4.34(m,2H),4.25(m,2H),4.17-4.11(m,2H),2.20-1.99(m,2H),1.91-1.76(m,2H),1.53-1.36(m,2H).

[0310] Example 6

[0311] 1-(5-Fluoro-6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-1,2-dihydropyridin-2-one

[0312] Step 1:

[0313] Compound 5a (3 g, 17.11 mmol), pyridin-2(1H)-one (813.49 mg, 8.55 mmol), pyridine (1.35 g, 17.11 mmol), copper acetate (3.1 g, 17.11 mmol), and 4A molecular sieves (3.25 g) were dissolved in DCM (60 mL) and DMF (5 mL) and reacted under an oxygen atmosphere for 6 h. The reaction was confirmed by TLC, filtered, and the filter cake was rinsed with dichloromethane. The combined organic phases were concentrated and purified by column chromatography to obtain compound 6a (1.8 g, yield: 93.68%).

[0314] MS m / z(ESI):225.1[M+H] +

[0315] Step 2:

[0316] Compound 4c (2.12 g, 2.85 mmol), compound 6a (640 mg, 2.85 mmol), cesium carbonate (13.92 g, 42.74 mmol), and dichloro[1,3-bis(2,6-di-3-pentylphenyl)imidazol-2-ylidene](3-chloropyridyl)palladium(II) (135.68 mg, 170.95 μmol) were weighed into a reaction flask and dioxane (60 mL) was added. Under nitrogen protection, the reaction was carried out at 110°C for 4 hours. LCMS analysis showed the formation of the target compound. The reaction solution was cooled to room temperature and filtered. The filtrate was evaporated to dryness, and water and ethyl acetate were added. The layers were separated, and the aqueous phase was extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by column chromatography to obtain compound 6 (350 mg, yield: 33.15%).

[0317] MS m / z(ESI):425.1[M+H] +

[0318] 1 H NMR (400MHz, DMSO-d6) δ7.86-7.80(m,2H),7.64(m,1H),7.56-7.43(m,2H),6.88(m,1H),6.75(m,1H),6.46(m,1H),6.28(m ,1H),4.47(m,1H),4.42-4.34(m,2H),4.23(m,1H),4.17-4.10(m,2H),2.09(m,2H),1.96-1.81(m,2H),1.65-1.37(m,2H).

[0319] Example 7

[0320] 1-(5-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyrazin-2-yl)-1,2-dihydropyridin-2-one

[0321] Step 1:

[0322] 2-Bromo-5-chloropyrazine (250 mg, 2.629 mmol), 2-hydroxypyridine (786 mg, 2.760 mmol), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (74.48 mg, 0.526 mmol), cuprous iodide (50.07 mg, 0.263 mmol), and potassium carbonate (726.66 mg, 5.258 mmol) were added to a reaction flask. DMSO (5 mL) was then added, and the atmosphere was purged with nitrogen three times. The reaction system was microwaved at 120°C for two hours. LC-MS confirmed complete conversion of the starting material. The product was concentrated under reduced pressure and purified on a silica gel column to afford compound 7a (390 mg, 59% yield).

[0323] MS m / z(ESI):252.1[M+1].

[0324] Step 2:

[0325] Compound 4c (100 mg, 0.191 mmol), compound 7a (72.17 mg, 0.286 mmol), Pd-PEPPSI-IPent catalyst (15.1 mg, 0.019 mmol), and cesium carbonate (310.72 mg, 0.954 mmol) were added to a reaction flask. 1,4-dioxane (1 mL) was then added, and the atmosphere was purged with nitrogen three times. The reaction system was stirred in a 100°C oil bath for 5 hours. LC-MS analysis confirmed complete conversion of the starting material. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The target compound 7 (10 mg) was obtained by silica gel column purification and reverse-phase HPLC.

[0326] MS m / z(ESI):408.5[M+1].

[0327] 1 H NMR (400MHz, DMSO) δ8.23(d,J=1.3Hz,1H),7.86–7.81(m,2H),7.73–7.68(m,1H),7.52–7.43(m,2H),6.79(d,J=7.2Hz,1H),6.46(dt,J=9.3,1.1Hz, 1H),6.32(td,J=6.7,1.4Hz,1H),4.42–4.36(m,2H),4.32–4.18(m,2H),4. 17–4.10(m,2H),2.20–2.00(m,2H),1.94–1.80(m,2H),1.55–1.43(m,2H).

[0328] Example 8

[0329] 2-(5-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyrazin-2-yl)-2,3-dihydro-1H-isoindol-1-one

[0330] Step 1:

[0331] Isoindolin-1-one (300 mg, 2.253 mmol), 2,5-dibromopyrazine (589 mg, 2.478 mmol), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (64.10 mg, 0.451 mmol), cuprous iodide (42.91 mg, 0.225 mmol), and potassium phosphate (956.53 mg, 4.506 mmol) were added to a reaction flask. 1,4-Dioxane (6 mL) was added, and the atmosphere was purged with nitrogen three times. The reaction system was stirred in a 100°C oil bath overnight. LCMS confirmed complete conversion of the starting material. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified on a silica gel column to afford compound 8a (270 mg, 41%).

[0332] MS m / z(ESI):190.0[M+1].

[0333] Step 2:

[0334] Compound 8a (60 mg, 0.210 mmol), compound 4c (100 mg, 0.191 mmol), Pd-PEPPSI-IPent catalyst (15.1 mg, 0.019 mmol), and cesium carbonate (310.72 mg, 0.954 mmol) were added to a reaction flask. 1,4-dioxane (1 mL) was then added, and the atmosphere was purged with nitrogen three times. The reaction system was stirred in a 100°C oil bath for 5 hours. LCMS analysis confirmed complete conversion of the starting material. The reaction solution was cooled to room temperature, concentrated under reduced pressure, and purified by silica gel column chromatography and Prep-HPLC to yield compound 8 (23 mg).

[0335] MS m / z(ESI):446.4[M+1].

[0336] 1H NMR (400MHz, DMSO) δ9.01(d,J=1.5Hz,1H),7.83(s,1H),7.81–7.76(m,2H),7.69–7.65(m,2H),7.57–7.51(m,1H),7.02(d,J=6.9Hz,1H),6.78(d ,J=7.2Hz,1H),4.98(s,2H),4.42–4.36(m,2H),4.28–4.18(m,2H),4.16 –4.11(m,2H),2.20–2.03(m,2H),1.93–1.77(m,2H),1.55–1.41(m,2H).

[0337] Example 9

[0338] 3-(6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-1-methyl-2-oxoylidetetrahydro-1H-imidazol-4-one

[0339] Step 1:

[0340] 2-Fluoro-5-iodopyridine 9a (5878 mg, 26.4 mmol), tert-butyl ((1S,3S)-3-aminocyclopentyl)carbamate (4400 mg, 21.97 mmol), DIEA (8518 mg, 65.91 mmol), and DMSO (60 mL) were placed in a reaction flask and reacted at 120°C under nitrogen for 16 h. LCMS confirmed the complete reaction. Ethyl acetate (200 mL) was added, and the mixture was washed three times with saturated brine (30 mL). The residue was concentrated under reduced pressure, and purified by column chromatography to obtain the target compound 9b (5000 mg, yield: 56.4%).

[0341] MS m / z(ESI):404.2[M+1].

[0342] Step 2:

[0343] Compound 9b (500 mg, 1.24 mmol), 1-methylimidazoline-2,4-dione (283 mg, 2.48 mmol), potassium phosphate (790 mg, 3.72 mmol), cuprous iodide (236 mg, 1.24 mmol), trans-N,N′-dimethylcyclohexane-1,2-diamine (212 mg, 1.50 mmol), and isopropanol (10 mL) were placed in a reaction flask and stirred at 110°C under nitrogen for 5 h. LCMS confirmed the formation of the product. The mixture was filtered through celite and the filtrate was concentrated under reduced pressure. The residue was added with deionized water and dichloromethane, and the layers were separated. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain compound 9c (250 mg, yield: 51.8%).

[0344] MS m / z(ESI):390.5[M+1].

[0345] Step 3:

[0346] Compound 9c (259 mg, 0.64 mmol) was dispersed in dichloromethane (5 mL), and 4N hydrochloric acid in dioxane (5 mL) was added. The mixture was allowed to react at room temperature for 4 h. LCMS confirmed complete reaction. The reaction solution was evaporated to dryness under reduced pressure and dissolved in a small amount of methanol. Ethyl acetate was added, and a large amount of solid precipitated. This solid was filtered and washed with ethyl acetate to obtain a crude product. This crude product was added to DCM (10 mL) in a reaction flask, and 1 mL of triethylamine was added. The reaction was stirred for 10 min, and the reaction solution was concentrated to obtain the crude product, compound 9d (180 mg).

[0347] MS m / z(ESI):290.5[M+1].

[0348] Step 4:

[0349] Compound 9d (180 mg, 0.62 mmol), compound 3b (108 mg, 0.62 mmol), cesium carbonate (2026 mg, 6.22 mmol), Pd-PEPPSI-IPent catalyst (99 mg, 0.12 mmol), and dioxane (10 mL) were weighed and reacted at 110°C under nitrogen for 16 hours. LCMS analysis revealed the product, which was filtered through celite. The filtrate was concentrated and analyzed by pre-HPLC to yield compound 9 (12 mg, 4.53% yield).

[0350] MS m / z(ESI):426.5[M+1].

[0351] 1H NMR (400MHz, DMSO-d6) δ7.85–7.81(m,2H),7.25(dd,J=8.9,2.6Hz,1H),6.84(d,J=6.9Hz,1H),6.75(d,J=7.2Hz,1H),6.50(d,J=8.9Hz,1H), 4.42–4.36(m,2H),4.28–4.16(m,2H),4.16–4.12(m,2H),4.07(s,2H), 2.90(s,3H),2.15–1.99(m,2H),1.89–1.73(m,2H),1.52–1.38(m,2H).

[0352] Example 10

[0353] 2-(6-{[(1S,3S)-3-(6,7-dihydro[1,4]dioxano[3,2-d]pyrimidin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-4-methyl-3H,2H-1,2-pyridazin-3-one

[0354] The compound of Example 10 was obtained by referring to the synthetic route of Example 1.

[0355] MS m / z(ESI):422.5[M+1].

[0356] 1H NMR(400MHz, DMSO-d6)δ8.08(d,J=2.4Hz,1H),7.89(d,J=4.0Hz,1H),7.83(s,1H), 7.49(dd,J=8.8,2.8Hz,1H),7.34(dd,J=4.0,1.2Hz,1H),6.87(d,J0=6.8Hz,1H),6 .76(d,J=7.2Hz,1H),6.51(dd,J=8.8,0.8Hz,1H),4.41–4.37(m,2H),4.34–4.17(m ,2H),4.16–4.11(m,2H),2.18–2.01(m,5H),1.90–1.76(m,2H),1.53–1.41(m,2H).

[0357] Example 11

[0358] 1-(6-{[(1S,3S)-3-(quinazolin-2-ylamino)cyclopentyl]amino}pyridin-3-yl)-1,2-dihydropyridin-2-one

[0359] The compound of Example 11 was obtained by referring to the synthetic route of Example 1.

[0360] MS m / z(ESI):399.19[M+1].

[0361] 1H NMR(400MHz,MeOD)δ9.02(s,1H),7.95(d,J=2.6Hz,1H),7.75(dd,J=8.1,1.5Hz,1H),7.70(ddd,J=8.4,6.9,1.5Hz, 1H),7.65–7.52(m,3H),7.45(dd,J=9.0,2.7Hz,1H),7.25(t,J=7.5Hz,1H),6.63(t,J=8.9Hz,2H),6.47(d,J=1.4Hz, 1H),4.65–4.54(m,1H),4.44–4.33(m,1H),2.39–2.27(m,2H),2.14–2.03(m,2H),1.75–1.59(m,2H).

[0362] Biological activity test

[0363] The present disclosure is further described and explained below in conjunction with test examples, but these embodiments are not intended to limit the scope of the present disclosure.

[0364] Test Example 1. Test of the binding capacity of the disclosed compounds to PCSK9

[0365] Table 1. Main experimental materials and instruments

[0366] 1. Experimental Procedure

[0367] 1.1 Reagent Preparation

[0368] 1) Protein: PCSK9 (biotin-labeled) (Acro).

[0369] The protein was dissolved in water to 200 μg / mL and stored at -80°C.

[0370] 2) Chip: SA chip.

[0371] 3) Running buffer: 1x HBSP + 4% DMSO + 0.1 mM CaCl2.

[0372] 1.2 Protein fixation

[0373] 1) Preheat the chip to room temperature;

[0374] 2) Prepare the required buffer and filter through a 0.22 μm filter membrane;

[0375] 3) Load the chip and write chip parameters;

[0376] 4) Dilute PCSK9 protein to 50 μg / mL with PBS;

[0377] 5) Setting up a fixed program in the BIAcore S200 instrument;

[0378] 6) Run a fixed program;

[0379] 7) The final fixed response value is 8000RU, and the maximum response value is 49.4RU.

[0380] 1.3 Compound Testing

[0381] 1) Dilute the compound in a 3-fold gradient using running buffer. Start with 0.4 μM for the reference compound and dilute 3-fold. Start with 10 μM for the test compound and dilute 3-fold.

[0382] 2) Prepare the dissolution correction buffer according to the table below:

[0383] Table 2. Lysis Correction Buffer

[0384] 3) Set the instrument's "LMW multi kinetics" program as the running program for compound testing.

[0385] a) The association time was set to 100 s and the dissociation time was set to 300 s.

[0386] 4) Run the compound test program.

[0387] 1.4 Data Analysis

[0388] 1) Rmax = (MW analyte / MW ligand)*RL*Sm

[0389] When using affinity fitting to fit the curve, the response value of the highest concentration should be close to the optimal Rmax value of the fit.

[0390] 2) Check solvent correction and compound concentration gradient.

[0391] 3) Check the baseline, binding, and positive compound results in the test report.

[0392] 4) Select the appropriate fitting method to fit the curve according to the actual situation.

[0393] 5) Check whether the fitting results pass the instrument data quality inspection standards.

[0394] Table 3. Binding affinity of the disclosed compounds to PCSK9 (K D )

[0395] From the results, the binding force of Examples 1-3 is stronger than that of Example 11, the binding force of Example 3 is more than 10 times stronger than that of Reference 2, and the binding force of Example 3 is comparable to that of Examples 7-8.

[0396] Reference 2 This is Example 487 in patent applications WO2020150473 and WO2020150474.

[0397] Test Example 2. Pharmacokinetics Experiment in Mice

[0398] C57BL6J mice were used as test subjects. LC / MS / MS was used to determine plasma drug concentrations at different times after oral administration of Reference 1 (WO2020150473 and WO2020150474, Example 458B) and the Examples. The pharmacokinetic behavior of Reference 1 and the Example compounds in mice was studied and their pharmacokinetic profiles were evaluated.

[0399] Experimental animals: 2 healthy 6-8 week old male mice (20-30 g) per group

[0400] Drug preparation: Weigh a certain amount of drug and prepare a 1 mg / mL colorless clear solution (solvent: 5% DMSO / 40% PEG400 / 55% normal saline).

[0401] Administration: The mice were fasted overnight and then gavage-administered. The dosage of reference substance 1 and the example compound was 10 mg / kg.

[0402] Procedure: Mice were orally administered with reference substance 1 and Example Compound. Approximately 0.03 mL of blood was collected by peripheral venipuncture at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after administration. The blood was placed in a tube containing EDTA-K2 and centrifuged at 4000 g per minute for 5 minutes at approximately 4°C to separate the plasma, which was then stored at -75°C.

[0403] Determination of the content of the test compound in mouse plasma after oral administration of different drug concentrations: Take 10 μL of mouse plasma at each time point after administration, add 5 μL of blank solution and 200 μL of acetonitrile solution containing internal standard dexamethasone, vortex mix for 30 seconds, centrifuge for 15 minutes (3900 rpm), and take 12 μL of the supernatant of the plasma sample for LC / MS / MS analysis.

[0404] Table 4. Pharmacokinetic parameter results of Reference 1 and Example 3

[0405] Note: The values ​​listed are the average values ​​of two animals. Reference 1 is Example 458B in patent applications WO2020150473 and WO2020150474.

[0406] From the results, Example 3 has better oral PK in mice, the blood drug exposure is nearly 3 times that of Reference 1, and the half-life is longer, indicating that the pharmacokinetic properties of the compound of the present invention are better than those of Reference 1.

[0407] Test Example 3. Effects of Reference Compounds and Example Compounds on LDLR Levels in HepG2 Cells

[0408] Table 5. Main experimental materials and instruments

[0409] Experimental procedures

[0410] HepG2 cells were seeded at 30,000 cells / well in a 96-well plate. The next day, cells were stimulated with 4 nM PCSK9 D374Y per well and treated with the test compound at the same concentration in triplicate. The plates were incubated at 37°C, 5% CO2 for 48 hours. The cells were washed twice with PBS, and the supernatant discarded. The cells were then placed on ice and treated with 50 μL / well of RIPA lysis buffer containing protease and phosphatase inhibitors. The cell culture plates were frozen at -80°C overnight. After thawing, they were lysed on ice for 30 minutes and centrifuged at low temperature for 15 minutes. The cell lysate was diluted 20-fold for ELISA analysis. 50 μL / well of sample or standard was added to the coated plates and incubated at room temperature for 2 hours. The plates were washed four times with wash buffer. 200 μL / well of human LDLR-conjugated antibody was added and incubated at room temperature for 2 hours. The wash step was repeated, and 200 μL / well of substrate solution was added. After 30 minutes, add 50 μL / well of stop solution and measure the OD value at 450 nm. Calculate the LDLR concentration in the sample wells based on the standard curve. The LDLR elevation ratio is calculated using the formula (sample value - mean value of control wells) / mean value of control wells. Data were processed using GraphPad Prism 8.

[0411] Table 6. Effects of Reference 1 and Example Compounds on LDLR Protein Levels in HepG2 Cells at a Concentration of 75 μM

[0412] Note: a Represents the average value of triplicate wells, and reference 1 is Example 458B in patent applications WO2020150473 and WO2020150474.

[0413] The same method was used to test the effects of the compounds of Examples 7 and 8 on the LDLR protein level in HepG2 cells at a concentration of 75 μM. The compound of Example 7 was 1.16 times that of reference substance 1, and the compound of Example 8 was 1.04 times that of reference substance 1.

[0414] From the results, reference substance 1 and compounds of Examples 3, 5, 6, 7 and 8 can all upregulate the LDLR level of HepG2 cells, among which the LDLR upregulation by compounds of Examples 3 and 5 is greater.

[0415] Test Example 4. Metabolite Test of Reference Material 1 and Example 3 Compounds in Mouse Hepatocytes

[0416] 1. Purpose of the experiment

[0417] The purpose of this study is to identify the possible metabolites of the test substance in different species of hepatocytes and to infer its possible metabolic pathways. The test concentration of the test substance is 10μM.

[0418] 2. Materials and Reagents

[0419] Mouse hepatocytes were stored in liquid nitrogen. Detailed information is shown in the table below.

[0420] 3. Experimental Design

[0421] 3.1 Preparation of compound working solution

[0422] The test substance and control drug verapamil powder were prepared into high concentration stock solutions with DMSO, and diluted with DMSO to 2 mM working solution before use. The final concentration of the test substance and verapamil was 10 μM.

[0423] 3.2 Preparation of hepatocytes

[0424] 1) Preheat the hepatocyte recovery solution and incubation solution in a 37°C water bath for at least 15 minutes before use.

[0425] 2) Take a vial of cryopreserved hepatocytes and ensure they remain frozen before thawing. Quickly place the hepatocytes in a 37°C water bath and gently shake until all ice crystals are dispersed. Spray with 70% ethanol and transfer to a biosafety cabinet.

[0426] 3) Pour the contents of the hepatocyte tubules into a centrifuge tube containing 50 mL of recovery medium and centrifuge at 100 g for 10 minutes. After centrifugation, aspirate the recovery medium and add enough incubation medium to obtain a cell density of approximately 1.5 × 10 6 cells / mL of cell suspension.

[0427] 4) Count the hepatocytes and determine the viable cell density using a Cellometer Vision. Dilute the hepatocyte suspension with incubation medium to a viable cell density of 1×10 6 cells / mL.

[0428] 3.3 Test methods

[0429] 1) Pipette 1 μL of a 2 mM solution of the test substance into a 24-well incubation plate, then add 199 μL of the suspended hepatocytes to initiate the reaction. Set the time points to 0, 2, and 4 hours, with different time points corresponding to different wells. Return the plate to the incubator (37°C, 5% CO2, 90-95% relative humidity) and incubate on a vortex at 500 rpm.

[0430] 2) After incubation for the corresponding time, immediately add 400 μL of cold acetonitrile containing 0.1% formic acid to terminate the reaction. Mix well and transfer to the corresponding EP

[0431] 3) Incubate and treat the control drug verapamil according to the above method, and only prepare samples at 0 and 4 hours. The remaining percentage of verapamil at 4 hours will be used to detect enzyme activity.

[0432] 4) All samples were vortexed for 100 seconds and then centrifuged at 16,000 g for 15 minutes to precipitate proteins. 60 μL of the supernatant was mixed with 60 μL of pure water and then analyzed by UHPLC-MS / MS.

[0433] 4. Instrument Configuration

[0434] A Vanquish ultra-high performance liquid chromatography system (Thermo Fisher Scientific, USA) was combined with a Thermo Scientific Q Exactive mass spectrometer (Thermo Fisher Scientific, USA) and equipped with a HESI ion source.

[0435] 5. Mass spectrometry detection

[0436] UHPLC-MS / MS data were acquired and processed using a Q Exactive high-resolution mass spectrometer. MS full scans were used to trigger MS / MS product ion scans using data-dependent acquisition (DDA).

[0437] The relevant mass spectrometry parameters of the test substance are optimized according to its own properties.

[0438] 6. Data Analysis

[0439] Data processing and analysis were performed using Xcalibur (v3.0 / 4.1, Thermo Fisher Scientific) software, Compound Discoverer 3.0 (Thermo Fisher Scientific) software, and Microsoft Excel 2016.

[0440] Table 7. Remaining proportion of parent drug of reference 1 and Example 3 (%)

[0441] Note: Reference 1 is Example 458B in patent applications WO2020150473 and WO2020150474

[0442] From the experimental results, it can be seen that the stability of the compound of Example 3 in mouse liver cells is superior to that of the reference compound 1.

[0443] Test Example 5. Stability test of test compound in hepatocytes

[0444] 1. Experimental Procedure

[0445] 1) Prepare high-concentration stock solutions of the test substance and control drug verapamil powder with DMSO, and dilute with DMSO to a 100 μM working solution before use. The final concentration of the test substance and verapamil is 1 μM.

[0446] 2) Take a vial of cryopreserved hepatocytes and ensure they remain frozen before thawing. Quickly place the hepatocytes in a 37°C water bath and gently shake until all ice crystals are dispersed. Spray with 70% ethanol and transfer to a biosafety cabinet.

[0447] 3) Pour the contents of the hepatocyte tubules from different species into a centrifuge tube containing 50 mL of recovery medium and centrifuge at 100 g for 10 minutes. After centrifugation, aspirate the recovery medium and add enough incubation medium to obtain a cell density of approximately 1.0 × 10 6 cells / mL of cell suspension.

[0448] 4) Count the hepatocytes and determine the viable cell density using a Cellometer Vision. The viability of the hepatocytes must be greater than 75%. Dilute the hepatocyte suspension with incubation medium to a viable cell density of 0.5 × 10 6 cells / mL.

[0449] 5) Transfer 198 μL of the live cell suspension to a 96-well deep-well plate. Place the plate on a vortex and preheat in an incubator for 10 minutes. Perform the experiment in duplicate.

[0450] 6) Add 2 μL of 100 μM test substance or verapamil to each well to initiate the reaction, and place the deep-well plate back on the incubator vortexer.

[0451] 7) Incubate the sample. At 0, 15, 30, 60, 90, and 120 minutes, aspirate 25 μL of the suspension and terminate the reaction by adding 150 μL of acetonitrile containing the internal standard. Vortex for 10 minutes and centrifuge at 3220 g at 4°C for 45 minutes. Transfer 100 μL of the supernatant to the injection plate and mix thoroughly with 100 μL of purified water for UPLC-MS / MS analysis.

[0452] 2. Data Analysis

[0453] All data calculations were performed using Microsoft Excel. Peak areas were determined by extracting ion spectra. The in vitro half-life (T) of the parent drug was determined by linearly fitting the natural logarithm of the parent drug elimination percentage with time. 1 / 2 ).

[0454] In vitro half-life (T 1 / 2 ) is calculated by slope:

[0455] In vitro T 1 / 2 =0.693 / k

[0456] In vitro clearance (μL / min / 10 6 cells) were calculated using the following formula:

[0457] In vitro CL int =kV / N

[0458] V = incubation volume per well (0.2 mL);

[0459] N = number of cells per well (0.1 × 10 6 cells)

[0460] Table 8. Metabolic clearance of test compounds in mouse hepatocytes

[0461] From the results, Example 3 has better stability in mouse hepatocytes than the compounds of Examples 1-2, Reference 1 and Reference 2.

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof, in, R 1 is selected from hydrogen or halogen; Ring A is selected from 3- to 7-membered cycloalkyl or 3- to 7-membered heterocycloalkyl, wherein the heterocycloalkyl contains at least one heteroatom selected from O, N or S; Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R A replace, Alternatively, two R attached to the same atom 2 Together with the connecting atom, it forms a 3- to 6-membered cycloalkyl or 3- to 6-membered heterocycloalkyl, wherein the 3- to 6-membered cycloalkyl or 3- to 6-membered heterocycloalkyl is optionally substituted by one or more R A replace, R A Selected from halogen, hydroxyl, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 Hydroxyalkyl; G1 is selected from -N- or -CR 3A -; G2 is selected from -N- or -CR 3B -; R 3A , R 3B and R 3 are each independently selected from hydrogen, halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl, wherein the heterocycloalkyl contains 1, 2 or 3 heteroatoms selected from O or N, -CONR'R" or -NH-C(O)-OR'; wherein C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl are optionally substituted with one or more R B Substitution, the R B Selected from halogen, hydroxyl, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy or 3- to 6-membered cycloalkyl; R' and R" are each independently selected from hydrogen, C 1-6 Alkyl or halogenated C 1-6 alkyl; Ring B is selected from 5- to 6-membered heterocycloalkyl or 5- to 6-membered heteroaryl, wherein the 5- to 6-membered heterocycloalkyl or 5- to 6-membered heteroaryl optionally contains at least one nitrogen atom and / or at least one oxo group; Each R 4 Each independently selected from halogen, hydroxyl, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl, the heterocycloalkyl containing 1, 2 or 3 heteroatoms selected from O or N, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy, 3 to 6 membered cycloalkyl or 4 to 10 membered heterocycloalkyl are optionally substituted with one or more R C Substitution, the R C Selected from halogen, hydroxyl, cyano, C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy or 3 to 6 cycloalkyl; m is selected from 0, 1, 2 or 3; n is selected from 0, 1, 2 or 3; and s is selected from 0, 1, 2 or 3.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is a 3- to 7-membered cycloalkyl group (eg, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cycloheptyl), preferably cyclopentyl.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is a 3- to 7-membered heterocycloalkyl group (e.g., 3-, 4-, 5-, 6-, or 7-membered), wherein the heterocycloalkyl group contains at least one heteroatom selected from O, N, or S; preferably, ring A is a 4- to 6-membered heterocycloalkyl group, wherein the heterocycloalkyl group contains one or two heteroatoms selected from O, N, or S; most preferably, ring A is selected from a 5- to 6-membered heterocycloalkyl group, wherein the heterocycloalkyl group contains one or two O atoms.

4. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, which is a compound represented by formula (I-1) or formula (I-2) or a pharmaceutically acceptable salt thereof, in, Ring B, R 1 , R 2 , G1, G2, R 3 , R 4 , m, n and s are as defined in claim 1.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein G1 is -N-; and G2 is -CR 3B -.

6. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, wherein G1 is -N-; and G2 is -N-.

7. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein G1 is -CR 3A -; and G2 is -CR 3B -.

8. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein Ring B is selected from Best 9. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, wherein Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R C Substitution, the R C is selected from halogen, hydroxyl or cyano; preferably, each R 4 Each is independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl.

10. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, wherein R 3A , R 3B and R 3 are each independently selected from hydrogen, halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R B Substitution, the R B is selected from halogen, hydroxyl or cyano; preferably, R 3A , R 3B and R 3 Each is independently selected from hydrogen, fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl.

11. A compound according to any one of claims 1 to 5 and 8 to 10, or a pharmaceutically acceptable salt thereof, which is a compound represented by formula (II-A-1), (II-A-2), (II-A-3), (II-A-4), (II-A-5) or a pharmaceutically acceptable salt thereof, in, R 1 , R 2 , R 3 , R 4 , m, n and s are as defined in claim 1.

12. The compound according to claim 11 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen; Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R A Substitution, the R A Selected from halogen, hydroxyl, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 Hydroxyalkyl; Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R B Substitution, the R B is selected from halogen, hydroxyl or cyano; Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R C Substitution, the R C is selected from halogen, hydroxyl or cyano; m, n and s are each independently selected from 0, 1, 2 or 3; Preferably, Each R 3 are each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl or trifluoromethoxy; Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl; m is 0; n and s are each independently selected from 0, 1 or 2.

13. A compound according to any one of claims 1 to 5 and 8 to 10, or a pharmaceutically acceptable salt thereof, which is a compound represented by formula (II-B-1), (II-B-2), (II-B-3), (II-B-4), (II-B-5) or a pharmaceutically acceptable salt thereof, in, R 1 , R 2 , R 3 , R 4 , m, n and s are as defined in claim 1.

14. The compound according to claim 13 or a pharmaceutically acceptable salt thereof, wherein R 1 is hydrogen; Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R A Replace, or attach two R 2 Together with the connecting carbon atom, it forms a 3- to 6-membered cycloalkyl group, wherein the 3- to 6-membered cycloalkyl group is optionally substituted by one or more R A replace, R A Selected from halogen, hydroxyl, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 Hydroxyalkyl; Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R B Substitution, the R B is selected from halogen, hydroxyl or cyano; Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R C Substitution, the R C is selected from halogen, hydroxyl or cyano; m, n and s are each independently selected from 0, 1, 2 or 3; or Preferably, Each R 2 Each independently selected from fluorine or methyl; or, two R connected to the same carbon atom 2 Together with the connecting carbon atom, it forms a cyclopropyl group, which is optionally substituted by one or more R A Substitution, the R A is a halogen; Each R 3 are each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl or trifluoromethoxy; Each R 4 each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropane methyl or cyanomethyl; m, n and s are each independently selected from 0, 1, 2 or 3.

15. A compound according to any one of claims 1 to 4, 6 and 8 to 10, or a pharmaceutically acceptable salt thereof, which is a compound represented by formula (II-C-1), (II-C-2), (II-C-3), (II-C-4) or a pharmaceutically acceptable salt thereof, in, R 1 , R 2 , R 3 , R 4 , m, n and s are as defined in claim 1.

16. The compound represented by formula (II-C-1) or (II-C-2) or a pharmaceutically acceptable salt thereof according to claim 15, wherein: R 1 is hydrogen; Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R A replace; R A Selected from halogen, hydroxyl, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 Hydroxyalkyl; Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R B Substitution, the R B is selected from halogen, hydroxyl or cyano; Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R C Substitution, the R C is selected from halogen, hydroxyl or cyano; m, n and s are each independently selected from 0, 1, 2 or 3; Preferably, Each R 3 Each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl, trifluoromethoxy; Each R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl; m is 0; n and s are each independently selected from 0, 1, 2 or 3.

17. The compound represented by formula (II-C-3) or (II-C-4) or a pharmaceutically acceptable salt thereof according to claim 15, wherein: R 1 is hydrogen; Each R 2 are each independently selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R A Substitution, or two R attached to the same carbon atom 2 Together with the connecting carbon atom, it forms a 3- to 6-membered cycloalkyl group, wherein the 3- to 6-membered cycloalkyl group is optionally substituted by one or more R A replace, R A Selected from halogen, hydroxyl, cyano, nitro, amino, carboxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl or C 1-6 Hydroxyalkyl; Each R 3 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 The alkoxy or 3 to 6 membered cycloalkyl group is optionally substituted with one or more R B Substitution, the R B is selected from halogen, hydroxyl or cyano; Each R 4 are independently selected from halogen, cyano, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-6 Alkoxy or 3 to 6-membered cycloalkyl, the C1-6 alkyl, C1-6 alkoxy, halogenated C1-6 alkyl, halogenated C1-6 alkoxy or 3 to 6-membered cycloalkyl optionally substituted by one or more R C Substitution, the R C is selected from halogen, hydroxyl or cyano; m, n and s are each independently selected from 0, 1, 2 or 3; Preferably, Each R 2 Each independently selected from fluorine or methyl; or two R connected to the same carbon atom 2 Together with the connecting carbon atom, it forms a cyclopropyl group, which is optionally substituted by one or more R A Substitution, the R A is a halogen; R 3 are each independently selected from fluorine, chlorine, cyano, methyl, trifluoromethyl or trifluoromethoxy; R 4 each independently selected from fluoro, chloro, cyano, methyl, trifluoromethyl, trifluoromethoxy, cyclopropyl or cyanomethyl; m, n, s are each independently selected from 0, 1, 2 or 3.

18. The compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, which is the following compound or a pharmaceutically acceptable salt thereof, Group A Group B Group C Group D 19. An isotope-substituted compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 18, wherein: The isotope substitution product is a deuterated product.

20. A pharmaceutical composition comprising the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof or the isotope substitution according to claim 19, and at least one pharmaceutically acceptable excipient.

21. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or an isotope substituted product according to claim 19, or a pharmaceutical composition according to claim 20, in the preparation of a medicament for preventing and / or treating diseases and conditions selected from dyslipidemia, dyslipoproteinemia, hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, hyperlipoproteinemia, xanthomas, hypoalphalipoproteinemia, sitosterolemia, atherosclerosis, arteriosclerosis, metabolic syndrome, coronary heart disease, peripheral vascular disease, congestive heart failure, stroke, vascular dementia, coronary artery disease, chronic kidney disease, retinopathy, inflammation, diabetic complications or thrombosis.

22. Use of the compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or the isotope substitution according to claim 19, or the pharmaceutical composition according to claim 20 in the preparation of a medicament for a PCSK9 inhibitor.

23. Use of a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, or an isotope substitute according to claim 19, or a pharmaceutical composition according to claim 20, in the preparation of a medicament for preventing and / or treating diseases and conditions mediated by PCSK9; preferably, the diseases and conditions mediated by PCSK9 are selected from dyslipidemia, dyslipoproteinemia, hypercholesterolemia, hyperlipidemia, hypertriglyceridemia, hyperlipoproteinemia, xanthomas, hypoalphalipoproteinemia, sitosterolemia, atherosclerosis, arteriosclerosis, metabolic syndrome, coronary heart disease, peripheral vascular disease, congestive heart failure, stroke, vascular dementia, coronary artery disease, chronic kidney disease, retinopathy, inflammation, diabetic complications or thrombosis.

24. A method for preparing a compound according to any one of claims 1 to 18 or a pharmaceutically acceptable salt thereof, comprising the steps of reacting a compound represented by formula (Ia) and a compound represented by formula (Ib) under alkaline conditions, optionally in the presence of a catalyst, LG is selected from halogen or -S(O)2R 7 , the R 7 Selected from C 1-6 Alkyl or halogenated C 1-6 Alkyl; and R 1 , R 2 , G1, G2, R 3 , R 4 , Ring A, Ring B, m, n and s are as defined in claim 1.

25. A compound represented by formula (Ib), in, LG is selected from halogen or -S(O)2R 7 , R 7 Selected from C 1-6 Alkyl or halogenated C 1-6 Alkyl; R 1 , R 2 and m is as defined in claim 1.

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