A nitrogen-containing heterocyclic compound and use thereof

Abstract

The application provides a kind of structurally novel nitrogen-containing heterocyclic compounds, its preparation method and pharmaceutical use, and provides a new direction for the development of PCSK9 inhibitor drugs. Experimental results show that the compounds of the application have high affinity for PCSK9 protein, can significantly reduce LDL-C, and can be used for treating cardiovascular diseases related to dyslipidemia. In vivo pharmacokinetic experiments show that the compounds of the application have good pharmacokinetic properties.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically, to a class of nitrogen-containing heterocyclic compounds, their preparation methods, and their pharmaceutical uses. Background Technology

[0002] High levels of low-density lipoprotein cholesterol (LDL-C) are a major risk factor for atherosclerosis and coronary heart disease. Currently, statins are the primary first-line drugs for lowering LDL-C; however, statins can be intolerable to some patients, or patients may not achieve the desired therapeutic effect at tolerated doses. Non-statin drugs, such as the cholesterol inhibitor ezetimibe, combined with statins can further lower LDL-C by 15-20%. Literature indicates that PCSK9 inhibitors combined with statins are even more effective in lowering LDL-C and can also overcome common statin side effects such as muscle pain.

[0003] PCSK9, short for Proprotein Convertase Subtilisin (Kexin type 9), is the 9th member of the subtilisin family of proprotein convertases and a protein closely related to cholesterol regulation. It was first reported in 2003. PCSK9 is primarily synthesized in the liver and contains three unique domains that play crucial roles in its biological function and intracellular transport. PCSK9 expression is regulated by various factors, such as SREBP2 (cholesterol regulatory element-binding protein 2). One of its main functions is the interaction between PCSK9 and the low-density lipoprotein receptor (LDLR). By binding to LDLR, it promotes its degradation within hepatocytes, thereby reducing the amount of LDLR on the hepatocyte surface and affecting the clearance of LDL-C (low-density cholesterol), potentially leading to elevated LDL-C levels in the blood and increasing the risk of cardiovascular disease. In addition to affecting LDLR, PCSK9 may also regulate other proteins associated with LDLR family members, such as ApoER2 (apolipoprotein E2 receptor) and VLDLR (very low density lipoprotein receptor), as well as other cell surface proteins such as CD36 and ACE2.

[0004] PCSK9 may play a key role in a variety of diseases, including cardiovascular disease, liver disease, infectious and autoimmune diseases, as well as neurocognitive impairment and cancer. Particularly in cardiovascular disease, PCSK9 increases disease risk by affecting cholesterol uptake. Furthermore, PCSK9 levels are significantly correlated with cholesterol, oxidized low-density lipoprotein (ox-LDL), and triglycerides.

[0005] Since the discovery of PCSK9 in 2003, various PCSK9 inhibitors have been developed and approved for marketing, including monoclonal antibodies (such as Evolocumab, Alirocumab, and Tafolecimab) and siRNAs (such as Inclisiran). These inhibitors reduce LDL-C levels in the blood by lowering PCSK9 levels, increasing the number of LDLRs on the surface of hepatocytes, and improving LDL-C clearance, demonstrating the potential to significantly reduce LDL-C and potentially reduce the risk of cardiovascular events. To date, there are no marketed oral small-molecule inhibitors of PCSK9. Two products (MK0616 and AZD0780) have shown relatively active clinical progress, while other small-molecule products have not disclosed significant clinical data in the past three years. Therefore, the development of oral PCSK9 inhibitors has significant research value and application prospects. Summary of the Invention

[0006] In a first aspect, the present invention provides a compound of formula (I), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0007] (I)

[0008] Ring A is selected from 5-10 member heterocyclic groups, C 6-10 Aryl or 5-10 heteroaryl groups; R a Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, -C(O)NR1R2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 cycloalkyl, phenyl, the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 Cycloalkyl or phenyl groups may be optionally replaced by one, two, or three groups selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-C 1-4 Alkyl, -C(O)-C 1-4 Substituents of alkyl groups; R1 and R2 are independently selected from hydrogen, deuterium, and C, respectively. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6Cycloalkyl, 4-6 membered heterocyclic groups, halogenated C 1-6 Alkyl, hydroxyl substituted C 1-6 alkyl; Ring B is selected from 9-16 membered bicyclic or tricyclic heterocyclic groups; R b Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, oxo, thio, =NH, =NC respectively. 1-4 Alkyl, =NC 3-6 cycloalkyl, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 Cycloalkyl, 5-6 membered heterocyclic, phenyl, 5-6 membered heteroaryl, wherein C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 The cycloalkyl, 5-6 membered heterocyclic, aryl, and 5-6 membered heteroaryl groups are optionally substituted by one, two, or three substituents selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, and cyano groups; or two R groups attached to the same atom. b The linkage forms a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally surrounded by 1, 2 or 3 groups selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl thiols, halogenated C 1-4 Alkyl, Halogenated C 1-4 Alkoxy, halogenated C 1-4 The substituents of the alkylthio group are replaced; R c Selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl thiols, halogenated C 1-4 Alkyl, Halogenated C 1-4 Alkoxy, halogenated C 1-4 Alkylthio; x is 0, 1, 2, 3 or 4; y is 0, 1, 2, 3 or 4; p can be 0, 1, 2, 3, or 4.

[0009] In a preferred embodiment of the present invention, ring A is selected from C.6-10 The ring is aryl or 5-10-membered heteroaryl; more preferably, ring A is selected from phenyl, 5-membered monocyclic heteroaryl, 6-membered monocyclic heteroaryl, 5-membered 6-membered heteroaryl, 6-membered 5-membered bicyclic heteroaryl, 5-membered 5-membered bicyclic heteroaryl, and 6-membered 6-membered bicyclic heteroaryl; even more preferably, ring A is selected from 5-membered monocyclic heteroaryl, 6-membered monocyclic heteroaryl, 5-membered 6-membered bicyclic heteroaryl, and 6-membered 5-membered bicyclic heteroaryl.

[0010] In a preferred embodiment of the present invention, ring A is selected from... , , , , , , . , , .

[0011] In a preferred embodiment of the present invention, ring A is selected from... .

[0012] In a preferred embodiment of the present invention, R a Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, -C(O)NR1R2, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 cycloalkyl, phenyl, the C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 Cycloalkyl or phenyl groups may be optionally replaced by one, two, or three groups selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-C 1-3 Alkyl, -C(O)-C 1-3 Substituents of alkyl groups; R1 and R2 are independently selected from hydrogen, deuterium, and C, respectively. 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 Cycloalkyl, 4-6 membered heterocyclic groups, halogenated C 1-3 Alkyl, hydroxyl substituted C 1-3 alkyl.

[0013] In a preferred embodiment of the present invention, R a Selected from C respectively 1-3 Alkyl, C 1-3 Alkoxy, the C 1-3 Alkyl, C 1-3The alkoxy group can be optionally replaced by one, two, or three halogens.

[0014] In a preferred embodiment of the present invention, R a The groups are selected from hydrogen, deuterium, halogen, cyano, -C(O)NR1R2, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylthio, ethylthio, n-propylthio, isopropylthio, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and phenyl, respectively. The methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylthio, ethylthio, n-propylthio, isopropylthio, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and phenyl groups are optionally substituted with 1, 2, or 3 substituents selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-CH3, and -C(O)-CH3. R1 and R2 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, oxacyclopentyl, halomethyl, haloethyl, halon-propyl, haloisopropyl, hydroxy-substituted methyl, hydroxy-substituted ethyl, hydroxy-substituted n-propyl, and hydroxy-substituted isopropyl; more preferably, one of R1 and R2 is selected from hydrogen, deuterium, methyl, and ethyl, and the other is selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, oxacyclopentyl, halomethyl, haloethyl, halon-propyl, haloisopropyl, hydroxy-substituted methyl, hydroxy-substituted ethyl, hydroxy-substituted n-propyl, and hydroxy-substituted isopropyl.

[0015] In a preferred embodiment of the present invention, R a The following groups are selected from hydrogen, F, Cl, Br, methyl, ethyl, methoxy, methylthio, -OCHF2, -OCH2F, -OCF3, monofluoromethyl, difluoromethyl, trifluoromethyl, cyclopropyl, cyclobutyl, hydroxymethyl, hydroxyethyl, hydroxy-substituted cyclopropyl, hydroxy-substituted cyclobutyl, cyano, -CH2-SO2-CH3, -CH2-CO-CH3, phenyl, -CONH2, -CONHCH3, -CON(CH3)CH2CH2OH, respectively. More preferably, -OCHF2 or trifluoromethyl.

[0016] In a preferred embodiment of the present invention, ring B is selected from 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 5-membered and 7-membered bicyclic heterocyclic groups, 7-membered and 5-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 7-membered bicyclic heterocyclic groups, 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 5-membered and 7-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups; more preferably, ring B is selected from 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 7-membered bicyclic heterocyclic groups.

[0017] In a preferred embodiment of the present invention, ring B is selected from... , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0018] In a preferred embodiment of the present invention, R bSelected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, oxo, thio, =NH, =NC respectively. 1-3 Alkyl, =NC 3-6 cycloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-6 cycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-6 The cycloalkyl group may be optionally substituted by one, two, or three substituents selected from deuterium, halogen, nitro, amino, hydroxyl, and cyano; or two R groups attached to the same atom. b The linkage forms a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally surrounded by 1, 2 or 3 groups selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 The substituents of the alkylthio group are replaced; In a preferred embodiment of the present invention, R b The radicals are selected from hydrogen, F, Cl, Br, amino, hydroxyl, cyano, oxo, thio, =NH, =N-CH3, =N-cyclopropane, =N-cyclobutane, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, methylthio, ethylthio, and cyclopropyl, wherein the methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, methylthio, ethylthio, cyclopropyl, and cyclobutane radicals are optionally substituted by 1, 2, or 3 substituents selected from halogens and cyano groups; or two R radicals attached to the same atom. b The links form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, azirne, azirne, cyclohexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, azirne, azirne are optionally substituted by 1, 2 or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, ethyl, methoxy, ethoxy; In a preferred embodiment of the present invention, R bThe radicals are selected from hydrogen, F, Cl, Br, cyano, oxo, thio, =NH, =N-CH3, =NH-cyclopropane, =N-cyclobutane, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, cyclopropyl; or two R radicals attached to the same atom. b The links form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, azirne, azirne, cyclohexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, azirne, azirne are optionally substituted by 1, 2 or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, methoxy.

[0019] In a preferred embodiment of the present invention Selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

[0020] In a preferred embodiment of the present invention, R c Selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; more preferably, R c Selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy; further preferably, R c Selected from hydrogen.

[0021] In a preferred embodiment of the present invention, x is 0, 1, 2 or 3; preferably, x is 0, 1 or 2.

[0022] In a preferred embodiment of the present invention, y is 0, 1, 2 or 3.

[0023] In a preferred embodiment of the invention, p is 0 or 1.

[0024] In a preferred embodiment of the invention, the compound represented by formula (I), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, is further represented by formula (II): (II) Each E is independently selected from CR d Or N, R d Independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; R b1 and R b2 The C atoms attached thereto form a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally composed of 1, 2 or 3 atoms selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 The substituents of the alkylthio group are replaced; Rings A and R a R cx and p are as described in equation (I).

[0025] In a preferred embodiment of the present invention, each E is a CR d Or, only one E is N, and the others are CR. d R d Independently selected from hydrogen, deuterium, halogen, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; more preferably, R d Independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; further preferably, R d Independently selected from hydrogen, deuterium, F, Cl, Br, cyano, methyl, and methoxy; further preferably, R d It is independently selected from hydrogen, F, Cl, and Br.

[0026] In a preferred embodiment of the present invention, R b1 and R b2 The C atoms connected thereto together form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, and azirnehexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, and azirnehexane are optionally substituted by 1, 2, or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, ethyl, methoxy, and ethoxy.

[0027] More preferably, R b1 and R b2 The C atoms connected thereto form cyclopropane, cyclobutane, oxecyclobutane, oxecyclopentane, oxecyclohexane, and azeocyclobutane, wherein the cyclopropane, cyclobutane, oxecyclobutane, oxecyclopentane, oxecyclohexane, and azeocyclobutane are optionally replaced by 1, 2, or 3 substituents selected from hydrogen, F, Cl, Br, and methyl.

[0028] In a preferred embodiment of the present invention Selected from or R b1 and R b2 The C atoms attached thereto form cyclopropane and aziridine, wherein the cyclopropane and aziridine are optionally substituted by one, two, or three substituents selected from hydrogen, F, Cl, Br, and methyl; R d It is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, and trifluoromethyl.

[0029] In a preferred embodiment of the invention, the compound represented by formula (I), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, is further represented by formula (III): (III) Each G is independently selected from CR g Or N, R g Independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; K is selected from -S-, -O-, -S(O)-, -S(O)2-, -C(R) k ) (R k )-、-S(O)(=NR k - or -N(R) k )-, where each R k Each element is independently selected from hydrogen, deuterium, halogen, cyano, and C. 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 cycloalkyl; Rings A and R a R b R c x, y, and p are as described in equation (I).

[0030] In a preferred embodiment of the present invention, each G is a CR g Or, only one G is N, and all others G are CR. g R g Independently selected from hydrogen, deuterium, halogen, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; more preferably, R g Independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; further preferably, R g Independently selected from hydrogen, deuterium, F, Cl, Br, cyano, methyl, and methoxy; further preferably, R g It is independently selected from hydrogen.

[0031] In a preferred embodiment of the present invention, K is selected from -S-, -O-, -S(O)-, -S(O)2-, and -C(R). k (R) k)-、-S(O)(NR k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, cyclopropyl, and cyclobutyl; more preferably, K is selected from -S-, -O-, -S(O)-, -S(O)2-, and -C(R). k ) (R k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, halogen, cyano, methyl, and methoxy; more preferably, K is selected from -S-, -O-, -S(O)2-, and -C(R). k ) (R k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, F, Cl, Br, and methyl; and even more preferably, K is selected from -S-, -O-, -S(O)2-, -CH2-, or -NH-.

[0032] In a preferred embodiment of the present invention, K is selected from -S-, -O-, -S(O)2- or -NH-, and G is always CR. g Or, only one G is N, and all others G are CR. g R g It is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, and trifluoromethyl.

[0033] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0034] The compounds described in this invention are selected from:

[0035] or its stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compound.

[0036] The compounds described in this invention are selected from:

[0037] or its stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compound.

[0038] The object of the present invention also includes providing a method for preparing compounds represented by the above general formula, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compounds.

[0039] The compound of the general formula can be prepared by a variety of methods, including but not limited to the following:

[0040] X1 is an amino protecting group, preferably selected from tert-butoxycarbonyl, benzyl, or 9-fluorenylmethoxycarbonyl; X2, X3, and X4 are independently selected from halogen, methyl mercapto, methanesulfonyl, borate, boric acid, or trifluoromethanesulfonyl, preferably chlorine, bromine, iodine, methyl mercapto, or boric acid; Ring A, Ring B, R a R b R c x, y, p are as described in compound (I).

[0041] Method a is a substitution or coupling reaction under basic conditions; method b is a halogenation reaction using a halogenating reagent; method c is a substitution or coupling reaction under basic conditions; method d is the removal of the amino protecting group under acidic, basic, or neutral conditions; method e is a substitution or coupling reaction under basic conditions.

[0042] The present invention also provides a pharmaceutical composition comprising the compound shown in the present invention, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound. Further, the pharmaceutical composition also comprises a pharmaceutically acceptable excipient.

[0043] The present invention also aims to provide the use of the compounds shown herein, stereoisomers, tautomers, or mixtures thereof, or pharmaceutically acceptable salts of the compounds, in the preparation of medicaments for treating and / or preventing PCSK9-mediated diseases. Preferably, PCSK9-mediated diseases are selected from cardiovascular diseases, liver diseases, infectious and autoimmune diseases, neurocognitive disorders, and cancer.

[0044] The object of the present invention also includes the use of the compounds shown in the present invention, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compounds in the preparation of medicaments for treating and / or preventing LDL reduction.

[0045] The object of the present invention also includes the use of the compounds shown in the present invention, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compounds in the preparation of medicaments for the treatment and / or prevention of cardiovascular diseases, liver diseases, infectious and autoimmune diseases, neurocognitive disorders and cancer.

[0046] definition

[0047] Unless otherwise specified, the term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, a straight-chain or branched group containing 1-20 carbon atoms, preferably containing 1-10 carbon atoms (i.e., C10). 1-10 Alkyl groups, more preferably containing 1-8 carbon atoms (C64- ... 1-8 Alkyl groups, more preferably containing 1-6 carbon atoms (i.e., C64-C ... 1-6 Alkyl), for example, "C 1-6 "Alkyl" refers to a group that is alkyl and has 1 to 6 carbon atoms in its carbon chain (specifically, 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, etc.

[0048] Unless otherwise specified, the term "cycloalkyl" refers to a hydrocarbon group selected from saturated cyclic hydrocarbon groups, including monocyclic and polycyclic (e.g., bicyclic and tricyclic) groups, including fused cycloalkyl, bridged cycloalkyl, or spirocycloalkyl. For example, a cycloalkyl group may contain 3 to 16 carbon atoms (such as 3 to 10, further such as 3 to 8, further such as 3 to 6, 3 to 5, or 3 to 4). Even further, for example, a cycloalkyl group may be selected from monocyclic groups containing 3 to 12 carbon atoms (such as 3 to 10, further such as 3 to 8, 3 to 6). Examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexadienyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl, and cyclododecyl. Examples of bicyclic cycloalkyl groups include those having 5 to 12, more such as 7 to 12 or 5 to 10 ring atoms arranged in a fused bicyclic arrangement selected from the [4,4], [4,5], [5,5], [5,6] or [6,6] ring systems, or arranged in a bridging bicyclic arrangement selected from bicyclic [2.2.1]heptane, bicyclic [2.2.2]octane and bicyclic [3.2.2]nonane. Other examples of bicyclic cycloalkyl groups include rings arranged in a bicyclic arrangement selected from the [5,6] and [6,6] ring systems.

[0049] Unless otherwise specified, the term "heterocyclic group" or "heterocycle" refers to a saturated or partially unsaturated monocyclic or polycyclic non-aromatic substituent having a ring carbon atom and 1 to 4 ring heteroatoms, comprising 3 to 20 ring atoms, wherein 1, 2, 3 or more ring atoms are selected from N, O or S, and the remaining ring atoms are C, wherein one or more S ring atoms (if present) and / or one or more N ring atoms (if present) may optionally be oxidized, wherein one or more carbon ring atoms may optionally be oxidized (i.e., to form an oxo group). Preferably, it comprises 3 to 16 ring atoms (3-16 membered heterocyclic group, preferably 3-14 membered heterocyclic group, more preferably 3-13 membered heterocyclic group, and even more preferably 3-12 membered heterocyclic group), and even more preferably 3-8 ring atoms (3-8 membered heterocyclic group), or 3-6 ring atoms (3-6 membered heterocyclic group), or 4-6 ring atoms (4-6 membered heterocyclic group), 5-8 ring atoms (5-8 membered heterocyclic group), or 5-6 ring atoms (5-6 membered heterocyclic group). The number of heteroatoms is preferably 1-4, more preferably 1-3 (i.e., 1, 2, or 3). Examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazoalkyl, tetrahydrofuranyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, pyranyl, etc. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups. A "heterocyclic group" can be a monocyclic ("monocyclic heterocyclic group") or a fused ("fused heterocyclic group" or "heterofused-cyclic group"), bridged ("heterobridged heterocyclic group" or "bridged-ring heterocyclic group") or spiro-fused ("heterospirocyclic group" or "spirocyclic heterocyclic group") ring system, such as a bicyclic system ("bicyclic heterocyclic group"), and can be saturated or partially unsaturated. A heterocyclic bicyclic system can include one or more heteroatoms in one or both rings. "Heterocyclic group" also includes a ring system in which the heterocyclic ring as defined above is fused with one or more carbocyclic groups, wherein the attachment point is on the carbocyclic or heterocyclic ring; or "heterocyclic group" also includes a ring system in which the heterocyclic ring as defined above is fused with one or more aryl or heteroaryl groups, or a ring system in which a cycloalkyl ring as defined above is fused with one or more heteroaryl groups, wherein the attachment point is on the heterocyclic or cycloalkyl ring, and in such cases, the number of members in the heterocyclic ring system is the number of atoms in the fused ring system. In some embodiments, each example of a heterocyclic group is independently optionally substituted, for example, unsubstituted (an "unsubstituted heterocyclic group") or substituted with one or more substituents (a "substituted heterocyclic group"). Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirropropyl, oxiranyl, and thiorenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirrocyclobutane, oxocyclobutane, and thiocyclobutane.Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrophenylthio, dihydrophenylthio, pyrrolidinyl, dihydropyrrolidinyl, and pyrrolidin-2,5-dione. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxolanecyclopentyl, oxathiocyclopentyl, dithiocyclopentyl, and oxazolidinyl-2-one. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithiocyclohexyl, and dioxolanecyclohexyl. Exemplary 6-membered heterocyclic groups containing 3 heteroatoms include, but are not limited to, triazacyclohexyl, oxadiazineyl, thiadiaziranyl, oxathiaziranyl, and dioxazinanyl. Exemplary 7-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azirheptanyl, oxadiazineyl, and thioheptanyl. Exemplary 8-membered heterocyclic groups containing 1 heteroatom include, but are not limited to, azirheptanyl, oxadiazineyl, and thioheptanyl. Exemplary 5-membered heterocyclic groups fused to a C6 aryl ring (also referred to herein as a 5,6-bicyclic heterocycle) include, but are not limited to, dihydroindolyl, isodihydroindolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, benzoxazolinone, etc. Exemplary 6-membered heterocyclic groups fused to an aryl ring (also referred to herein as a 6,6-bicyclic heterocycle) include, but are not limited to, tetrahydroquinolinyl, tetrahydroisoquinolinyl, etc.

[0050] "Alkoxy" refers to -O-alkyl, and the alkyl group is defined as above, i.e., containing 1-20 carbon atoms, preferably 1-10 carbon atoms, more preferably 1-8 carbon atoms, and even more preferably 1-6 carbon atoms (specifically 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, 1-methylpropoxy, 2-methylpropoxy, tert-butoxy, pentoxy, 1-methylbutoxy, 2-methylbutoxy, 3-methylbutoxy, 1,1-dimethylpropoxy, 1,2-dimethylpropoxy, 2,2-dimethylpropoxy, 1-ethylpropoxy, etc.

[0051] Unless otherwise specified, the term "alkoxy" refers to the form in which the oxygen in "alkoxy" is replaced by sulfur.

[0052] Unless otherwise specified, the terms "halogen" or "halogenated" refer to F, Cl, Br, and I. The term "halogenated alkyl" refers to an alkyl group as defined above in which one, two, or more hydrogen atoms, or all hydrogen atoms, are replaced by a halogen. Representative examples of halogenated alkyl groups include CCl3, CF3, CHCl2, CH2Cl, CH2Br, CH2I, CH2CF3, and CF2CF3.

[0053] Unless otherwise specified, the term "aryl" refers to an aromatic carbocyclic system containing 6-16 carbon atoms, or 6-14 carbon atoms, or 6-12 carbon atoms, or 6-10 carbon atoms, preferably 6-10 carbon atoms. The term "aryl" may be used interchangeably with the term "aromatic ring." Examples of aryl groups may include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthryl, or pyreneyl.

[0054] Unless otherwise specified, the term "heteroaryl" refers to an aromatic monocyclic, bicyclic, or polycyclic cyclic system containing a 5-16 member structure, or a 5-14 member structure, a 5-12 member structure, a 5-10 member structure, a 5-8 member structure, or a 5-6 member structure, wherein one, two, three, or more ring atoms are heteroatoms and the remaining atoms are carbon atoms, the heteroatoms being independently selected from O, N, or S, and the number of heteroatoms is preferably one, two, or three. Examples of heteroaryl groups may include, but are not limited to, furanyl, thiophene, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrroloyl, pyrazolyl, imidazoleyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purine, indoleyl, isoindoleyl, indazoleyl, benzofuranyl, benzothiophene, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzoimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazo[1,2-a]pyridyl, and pyrazolo[1,5-a]. Pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridinyl, etc.

[0055] Unless otherwise specified, the terms "pharmaceutically acceptable salt" or "medicinal salt" refer to a salt that, within reasonable medical judgment, is suitable for contact with mammalian, particularly human, tissues without excessive toxicity, irritation, allergic reactions, etc., and is proportionate to a reasonable benefit / risk ratio. Medically acceptable salts of amines, carboxylic acids, and other types of compounds are well known in the art. The salts can be prepared in situ during the final isolation and purification of the compounds of this invention, or solely by reacting a free base or free acid with a suitable reagent.

[0056] The compounds of this invention also include their "isotope derivatives." Unless otherwise specified, the term "isotope derivative" refers to compounds of this invention that can exist in an isotopically traced or enriched form, containing one or more atoms whose atomic weights or mass numbers differ from the atomic weights or mass numbers of the most abundant atoms found in nature. Isotopes can be radioactive or non-radioactive isotopes. Commonly used isotopes for isotopic labeling are: hydrogen isotopes, 2 H and 3 H; Carbon isotopes: 13 C and 14 C; Chlorine isotopes: 35 Cl and 37 Cl; Fluorine isotopes: 18 F; Iodine isotopes: 123 I and 125 I; Nitrogen isotopes: 13 N and 15 N; oxygen isotopes: 15 O, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. Especially 3 H and 13 C, because they are easy to label and convenient to detect, are more widely used. Some heavy isotopes, such as deuterium (… 2 Substitution with H can enhance metabolic stability and prolong the half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthetic techniques, just like non-isotope-labeled compounds.

[0057] The compounds of this invention also include their solvates. Unless otherwise specified, the terms "solvate" or "solvent" refer to the physical association of the compound of this invention with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. The solvent molecules in the solvate may be present in a regular and / or disordered arrangement. The solvate may contain stoichiometric or non-stoichiometric solvent molecules. "Solvate" encompasses both solution phases and separable solvates. Solventization methods are well known in the art.

[0058] Unless otherwise specified, the term "stereoisomer" refers to compounds having the same chemical structure but with different spatial arrangements of atoms or groups. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and inhibited isomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0059] Unless otherwise specified, the term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (also called proton transfer tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons.

[0060] The compounds of this invention also include their prodrugs. Unless otherwise specified, the term "prodrug" refers to a drug that is converted into a parent drug in vivo. Prodrugs are generally useful because they can improve certain, undesirable physical or biological properties. Physical properties are generally related to solubility (excessive or insufficient lipid or water solubility) or stability, while problematic biological properties include metabolism that is too rapid or poor bioavailability, which may itself be related to physicochemical properties. For example, they can be bioavailable orally, whereas the parent drug cannot. Prodrugs also have improved solubility in pharmaceutical compositions compared to the parent drug. An example of a prodrug, but not limited thereto, can be any compound of this invention administered as an ester ("prodrug") to facilitate transmembrane transport, where water solubility is detrimental to migration but beneficial once inside the cell, and which is subsequently metabolized and hydrolyzed into a carboxylic acid, i.e., the active entity. Another example of a prodrug can be a short peptide (polyamino acid) bound to an acid group, where the peptide is metabolized to exhibit the active moiety.

[0061] Unless otherwise specified, the term "optional substitution" means that the hydrogen at the substituted site of the group is not substituted, or is substituted by one or more substituents, preferably by one, two, or three substituents, wherein the substituents are preferably selected from the group consisting of: halogen, hydroxyl, mercapto, cyano, nitro, amino, azide, oxo, carboxyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 cycloalkyl, C 3-10 Cycloalkylsulfonyl, 3-10 membered heterocyclic group, C 6-14Aryl or 5-10 membered heteroaryl rings, wherein the C 2-6 Alkenyl, C 2-6 alkynyl group, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 cycloalkyl, C 3-10 Cycloalkylsulfonyl, 3-10 membered heterocyclic group, C 6-14 The aryl or 5-10 membered heteroaryl group may optionally be selected from halogen, hydroxyl, amino, cyano, C 1-6 Alkyl or C 1-6 One or more substituents in the alkoxy group are substituted, wherein the oxo group refers to two H atoms at the same substitution position being replaced by the same O atom to form a double bond.

[0062] The beneficial effects of this invention are as follows: This invention designs a class of novel compounds, providing a new direction for the development of PCSK9 inhibitor drugs. Experimental results show that the compounds of this application have a high affinity for PCSK9 protein, can significantly reduce LDL-C, and can be used to treat diseases including cardiovascular diseases related to dyslipidemia. In vivo pharmacokinetic experiments demonstrate that the compounds of this invention have favorable pharmacokinetic properties. Detailed Implementation

[0063] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of the present invention. The preferred embodiments and materials shown herein are for illustrative purposes only.

[0064] The structures of the compounds in this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS) and / or high-performance liquid chromatography (HPLC). The NMR measurements were performed using a Bruker AVANCE III 600 MHz instrument; the LC-MS measurements were performed using an LCMS WATERS ACQUITY UPLC H-Class PLUS and / or SQD2 instrument; and the HPLC measurements were performed using a WATERS e2695_2998 and / or an Agilent 1100 instrument.

[0065] The starting materials used in the embodiments of the present invention are known and commercially available, or can be synthesized using or in accordance with methods known in the art.

[0066] Example 1: Synthesis of spiro[cyclopropane-1,3'-indoline]-2'-one

[0067] In a 100 mL three-necked flask, indole-2-one (2 g), THF (30 mL), and DIPEA (4.0 g) were added. Under a nitrogen atmosphere, the system was cooled to -40 °C, and a 2.5 M solution of n-butyllithium hexane (27 mL) was added dropwise. After the addition was complete, the reaction was maintained at this temperature for 1 h. Then, the temperature was raised to 0 °C, and a solution of 1,2-dibromoethane (8.5 g) in THF (10 mL) was added dropwise. After the addition was complete, the reaction system was allowed to return to room temperature overnight. The reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, and the organic phase was concentrated to dryness under reduced pressure. The concentrate was purified by column chromatography (PE:EA = 2:1) to give the product (0.7 g). ESI-MS (m / z): 160.07 [M+H] + .

[0068] Intermediate Preparation Example 2: 3,4-Dihydrobenzo[ f [1,4]sulfur-nitrogen-5(2 H Synthesis of )-ketones

[0069] In a 100 mL reaction flask, methyl 2-mercaptobenzoate (3.00 g), 2-chloroethylamine hydrochloride (2.07 g), DMF (10 mL), and THF (10 mL) were added. Sodium hydride (2.22 g) was slowly added at 0 °C. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic phase was concentrated under reduced pressure, slurried with a mixed solution (EA:PE = 1:1), and filtered to obtain the product (1.30 g). ESI-MS (m / z): 180.05 [M+H] + .

[0070] Example 3: Synthesis of 1-methylspiro[azacyclobutane-3,3'-indoline]-2'-one

[0071] Spiro[azacyclobutane-3,3'-indoline]-2'-one (30 mg), methanol (5 mL), paraformaldehyde (15 mg), and sodium cyanoborohydride (21 mg) were added to a 25 mL reaction flask and stirred at room temperature for 4 h. Water (approximately 1 mL) was added to the reaction system, and the mixture was concentrated to dryness under reduced pressure. The concentrate was separated by preparative thin-layer chromatography (DCM:MeOH = 10:1) to obtain the product (20 mg). ESI-MS (m / z): 189.09 [M+H] + .

[0072] Intermediate Preparation Example 4: 3,4-Dihydrobenzo[ f [1,4]sulfur-nitrogen-5(2 H )-Ketone 1,1-dioxide

[0073] 3,4-Dihydrobenzo[f][1,4]thiazazon-5(2H)-one (1.00 g), m-chloroperoxybenzoic acid (3.85 g), and DCM (10 mL) were added to a 100 mL reaction flask and reacted at room temperature for 1 h. The product (0.61 g) was obtained by direct filtration. ESI-MS (m / z): 212.03 [M+H] + .

[0074] Example 5 of intermediate preparation: (1) S ,3 S )- N 1 -(5-(difluoromethoxy)pyrimidin-2-yl)- N 3 -(5-iodopyridin-2-yl)cyclopentane-1,3-diamine

[0075] Step 1: Add 2-chloro-5-(difluoromethoxy)pyrimidine (2 g) and DMSO (10 mL) to a 250 mL reaction flask, then add (1 g) of DMSO. S ,3 S 3-Aminocyclopentylcarbamate tert-butyl ester (2.7 g), DIPEA (4.5 g). The reaction solution was heated to 100 °C and reacted for 5 h. Water and ethyl acetate were added, the mixture was stirred, and the layers were separated. The resulting organic phases were washed once with water and once with saturated brine. The organic phases were concentrated to dryness under reduced pressure, and the concentrate was slurried with n-hexane to give the product (3.5 g). ESI-MS (m / z): 345.17 [M+H] + .

[0076] Step 2: Add (1) to a 250 mL reaction flask sequentially. S ,3 S 3.5 g of tert-butyl 3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentylcarbamate (30 mL) was reacted with 1,4-dioxane hydrochloride (4 M) at room temperature for 4 h. The reaction solvent was concentrated to dryness under reduced pressure, the concentrate was diluted with methanol, and then Ambersep® 900 resin was added under stirring to adjust the pH of the solution to approximately 8-9. The solution was filtered, and the filtrate was concentrated to dryness under reduced pressure to obtain the product (2.2 g). ESI-MS (m / z): 245.11 [M+H] +.

[0077] Step 3: In a 250 mL reaction flask, sequentially (1 S ,3 S )- N 1 -(5-(difluoromethoxy)pyrimidin-2-yl)cyclopentane-1,3-diamine (2.2 g), DMSO (10 mL), potassium carbonate (3.8 g), and 2-fluoro-5-iodopyridine (2.1 g) were added. The reaction mixture was heated to 110 °C and reacted for 16 h. Water and ethyl acetate were added, the mixture was stirred, and the layers were separated. The resulting organic phases were washed once with water and once with saturated brine. The organic phases were concentrated to dryness under reduced pressure, and the concentrate was purified by column chromatography (PE:EA = 3:1) to give the product (3.5 g). ESI-MS (m / z): 448.04 [M+H] + .

[0078] Example 6 of intermediate preparation: (1) S ,3 S )- N 1 -(5-(trifluoromethyl)pyrimidin-2-yl)- N 3 -(5-iodopyridin-2-yl)cyclopentane-1,3-diamine

[0079] The synthesis method is the same as in Example 5 for intermediate preparation, except that the starting material 2-chloro-5-(difluoromethoxy)pyrimidine in the first step is replaced with 2-chloro-5-(trifluoromethyl)pyrimidine, yielding (1 S ,3 S )- N 1 -(5-(trifluoromethyl)pyrimidin-2-yl)- N 3 1,3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (600 mg). ESI-MS (m / z): 450.03 [M+H] +

[0080] Intermediate Preparation Example 7: 3,4-Dihydropyrido[3,4-f][1,4]thiazide-5(2 H )-ketone

[0081] Step 1: Methyl 4-chloronicotinic acid (8 g, 46.67 mmol) was added to DMF (80 mL). K₂CO₃ (7.54 g) and tert-butyl (2-mercaptoethyl)carbamate (12.42 g) were added sequentially to the reaction solution. The reaction solution was stirred at 110 °C for 3 h. The reaction solution was extracted with water and ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The product (1.6 g) was obtained by column chromatography (PE:EA = 1:1). ESI-MS (m / z): 313.13 [M+H] + .

[0082] Step 2: 4-((2-(tert-Butoxycarbonyl)amino)ethyl)thio)nicotinic acid methyl ester (7.1 g) was dissolved in DCM (71 mL) and trifluoroacetic acid solution (71 mL). The mixture was stirred at 20°C for 2 h. The product (7.2 g) was concentrated under reduced pressure. ESI-MS (m / z): 213.02 [M+H] +

[0083] Step 3: 4-((2-aminoethyl)thio)nicotinic acid methyl ester (7.2 g) was dissolved in THF (72 mL) and MeOH (72 mL). Sodium methoxide (9.31 g) was added to the reaction solution at 0°C, and the mixture was stirred at 48°C for 16 h. After removing the solvent by vacuum distillation, the reaction solution was extracted with water (120 mL) and DCM:MeOH (3:1). The organic phase was dried over anhydrous sodium sulfate and then distilled under vacuum to obtain the product (3.9 g). ESI-MS (m / z): 181.03 [M+H] + .

[0084] Example 1: 1'-(6-(((1) S ,3 S Synthesis of 3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)spiro[cyclopropane-1,3'-indoline]-2'-one (compound a2)

[0085] Add (1) to a 50 mL reaction flask sequentially. S ,3 S )- N 1 -(5-(difluoromethoxy)pyrimidin-2-yl)- N 3-(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (100 mg), spiro[cyclopropane-1,3'-indoline]-2'-one (45 mg), cesium carbonate (220 mg), cuprous iodide (42 mg), (1 S ,2 S )- N 1 , N 2 -Dimethylcyclohexane-1,2-diamine (31 mg), DMSO (2 mL), reacted at 120 °C for 8 h under nitrogen protection. Water and ethyl acetate were added, the mixture was stirred, and the layers were separated. The resulting organic phases were washed once with water and once with saturated brine. The organic phases were concentrated to dryness under reduced pressure, and the concentrate was purified by column chromatography (DCM: MeOH = 30: 1) to give the product (80 mg). ESI-MS (m / z): 479.19 [M+H] + . 1 H NMR (600 MHz, CD3OD) δ 8.17 (s, 2H), 8.00 (s, 1H), 7.44 (d, J = 8.8 Hz, 1H), 7.20 (t, J = 7.7 Hz, 1H), 7.07 (t, J = 7.5 Hz, 1H), 7.01 (d, J = 7.4 Hz, 1H), 6.79 (d, J = 7.8 Hz, 1H), 6.70 (s, 1H), 6.69 (d, J = 73.5 Hz, 1H), 4.45-4.30 (m, 2H), 2.36–2.23 (m, 2H), 2.14–1.99 (m, 2H), 1.77-1.69 (m, 4H), 1.67-1.60 (m, 2H).

[0086] Example 2: 1'-(6-(((1) S ,3 S Synthesis of 3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-6'-fluorospiro[cyclopropane-1,3'-indoline]-2'-one (compound a1)

[0087] The synthesis method was the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one was replaced with 6'-fluorospiro[cyclopropane-1,3'-indoline]-2'-one. The product was obtained (33 mg). ESI-MS (m / z): 497.19 [M+H] + . 1 H NMR (600MHz, CD3OD) δ 8.15 (s, 2H), 7.97 (d, J = 2.4 Hz, 1H), 7.41 (dd, J = 8.9, 2.5Hz, 1H), 6.97 (dd, J = 8.2, 5.1 Hz, 1H), 6.79 – 6.74 (m, 1H), 6.67 (t, J =73.2 Hz, 1H), 6.66 (d, J = 8.9 Hz, 1H), 6.53 (dd, J = 9.1, 2.1 Hz, 1H), 4.43 –4.28 (m, 2H), 2.31 – 2.17 (m, 2H), 2.08 – 1.94 (m, 2H), 1.75 – 1.65 (m, 4H),1.64 – 1.54 (m, 2H).

[0088] Example 3: 1'-(6-(((1) S ,3 S Synthesis of 3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)spiro[azacyclobutane-3,3'-indoline]-2'-one (compound a58)

[0089] Step 1: Add (1) to a 50 mL reaction flask in sequence. S ,3 S )- N 1 -(5-(difluoromethoxy)pyrimidin-2-yl)- N 3 -(5-iodopyridin-2-yl)cyclopentane-1,3-diamine (100 mg), 2-oxaspiro[1H-indole-3,3'-azacyclobutane]-1'-carboxylic acid tert-butyl ester (77 mg), cesium carbonate (220 mg), cuprous iodide (42 mg), (1 S ,2 S )- N 1 , N2 -Dimethylcyclohexane-1,2-diamine (31 mg), DMSO (2 mL), reacted at 120 °C for 8 h under nitrogen protection. Water and ethyl acetate were added, the mixture was stirred, and the layers were separated. The resulting organic phases were washed once with water and once with saturated brine. The organic phases were concentrated to dryness under reduced pressure, and the concentrate was purified by column chromatography (DCM: MeOH = 30:1) to give the product (70 mg). ESI-MS (m / z): 594.26 [M+H] +

[0090] Step 2: Add 1'-(6-(((1) to a 100mL reaction flask S ,3 S 40 mg of tert-butyl 2'-oxospiro[azacyclobutane-3,3'-indoline]-1-carboxylic acid was dissolved in dioxane (10 mL) and slowly added dropwise to dioxane hydrochloride solution (2 M, 10 mL). The mixture was stirred at room temperature for 3 h, then water and ethyl acetate were added, and the mixture was stirred. The organic phases were separated and washed once with water and once with saturated brine. The organic phases were concentrated to dryness under reduced pressure, and the concentrate was purified by column chromatography (DCM: MeOH = 30 : 1) to give the product (36 mg). ESI-MS (m / z): 494.19 [M+H] + . 1 H NMR (600 MHz, CDCl3) δ 8.18 (s, 2H), 8.12 (s, 1H), 7.97 (d, J = 7.2 Hz, 1H), 7.43 (dd, J = 8.8, 2.4 Hz, 1H), 7.26 – 7.23 (m, 1H), 7.22 –7.18 (m, 1H), 6.74 (d, J = 1.2 Hz, 1H), 6.49 (d, J = 8.8 Hz, 1H), 6.41 (t, J = 73.2 Hz, 1H), 5.33 (d, J = 6.8 Hz, 1H), 4.89 (d, J = 6.4 Hz, 1H), 4.43 (d, J = 7.1 Hz, 3H), 4.26 (dd, J = 11.7, 5.5 Hz, 1H), 4.08 (d, J= 8.0 Hz, 2H), 2.40 – 2.29 (m, 3H), 2.12 – 2.01 (m, 2H), 1.64 – 1.54 (m, 2H).

[0091] Example 4: 1'-(6-(((1) S ,3 S Synthesis of 3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-1-methylspiro[azacyclobutane-3,3'-indoline]-2'-one (a59)

[0092] The synthesis method was the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one was replaced with 1-methylspiro[azacyclobutane-3,3'-indoline]-2'-one. The product was obtained (35 mg). ESI-MS (m / z): 508.20 [M+H] + . 1 H NMR (600 MHz, CDCl3) δ 8.18 (s, 2H), 8.11 (s, 1H), 7.84 (d, J = 5.4 Hz, 1H), 7.42(d, J =7.2 Hz, 1H), 7.24-7.12 (m, 2H), 6.74 (d, J = 6.9 Hz, 1H), 6.48 (d, J =8.4 Hz, 1H), 6.41 (t, J = 72.6Hz, 1H),5.26 (s, 1H), 4.81 (s, 1H), 4.43 (d, J = 5.6 Hz, 1H), 4.27 (s, 1H), 3.71 (s, 4H), 2.54 (s, 3H), 2.34 (s, 2H), 2.05(d, J = 6.8 Hz, 2H), 1.59 (s, 2H).

[0093] Example 5: 4-(6-(((1) S, 3 S )-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[f][1,4]thiazazide-5(2 H Synthesis of )-ketone (compound a64)

[0094] The synthesis method is the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one is replaced with 3,4-dihydrobenzo[ f [1,4]sulfur-nitrogen-5(2 H )-ketone, yielding the product (61 mg). ESI-MS (m / z): 499.19 [M+H] + . 1 H NMR (600MHz, DMSO -d6 ) δ 8.24 (s, 2H), 7.98 (s, 1H), 7.70 – 7.30 (m, 6H), 7.03 (t, J =74.0 Hz, 1H), 6.77 (d, J = 4.7 Hz, 1H), 6.52 (d, J = 8.6 Hz, 1H), 4.30 (dd, J = 13.4, 6.5 Hz, 2H), 3.67 (s, 2H), 3.15 (s, 2H), 2.22 – 2.04 (m, 2H), 1.95 –1.82 (m, 2H), 1.60 – 1.40 (m, 2H).

[0095] Example 6: Synthesis of 6-bromo-1-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one (a24)

[0096] The synthesis method was the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one was replaced with 6-bromo-2',3',5',6'-tetrahydrospiro[indoline-3,4'-pyran]-2-one, yielding the product (30 mg). ESI-MS (m / z): 601.45 / 603.45 [M+H] + . 1 H NMR (600 MHz, CDCl3) δ 8.18 (s, 2H), 8.08 (d, J = 2.1 Hz, 1H), 7.39 (dd, J = 8.8, 2.5 Hz, 1H), 7.27 (d, J = 2.5 Hz, 1H), 7.23 (dd,J = 7.9, 1.5 Hz, 1H), 6.90 (d, J = 1.4 Hz, 1H), 6.50 (d, J = 8.8 Hz, 1H), 6.41 (t, J =73.2Hz, 1H), 5.24 (d, J = 6.8 Hz, 1H), 4.84 (d, J = 6.5 Hz, 1H), 4.43 (dd, J = 13.7, 6.8 Hz, 1H), 4.27 (dt, J = 12.1, 5.8 Hz, 3H), 3.93 (dt, J = 11.6, 4.5Hz, 2H), 2.35 (td, J = 12.1, 4.8 Hz, 2H), 2.13 – 2.01 (m, 2H), 1.95 – 1.94(m, 2H), 1.65 -1.63 (m, 2H), 1.61– 1.56(m, 2H).

[0097] Example 7: 1'-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)spiro[cyclopropane-1,3'-pyrrolo[3,2-] c ]pyridine]-2'(1' H Synthesis of )-ketone (compound a8)

[0098] The synthesis method is the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one is replaced with spiro[cyclopropane-1,3'-pyrrolo[3,2-] c ]pyridine]-2'(1' H The product (28 mg) was obtained by reacting the ketone. ESI-MS (m / z): 480.50 [M+H] + . 1 HNMR (600 MHz, CD3OD) δ 9.16 (s, 1H), 8.99 (s, 2H), 8.79 (s, 1H), 8.40-7.94(m, 2H),7.70 (t, J = 78.0Hz, 1H), 7.61 (s, 1H), 7.40 (d, J= 7.3 Hz, 1H), 3.95(s, 2H), 2.78-2.68 (m, 2H), 2.64 (s, 2H), 2.55-2.41 (m, 2H), 2.39-2.22 (m, 2H), 2.05-1.98(m, 2H).

[0099] Example 8: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[ f [1,4]sulfur-nitrogen-5(2 H Synthesis of 1,1-ketodioxide (compound a68)

[0100] The synthesis method is the same as in Example 1, except that spiro[cyclopropane-1,3'-indoline]-2'-one is replaced with 3,4-dihydrobenzo[ f [1,4]Thiazine-5(2 H )-ketone 1,1-dioxide was reacted to give the product (26 mg). ESI-MS (m / z): 531.55 [M+H] + . 1 HNMR (600 MHz, CD3OD) δ 8.17 (s, 2H), 8.09 (s, 1H), 8.05 (d, J = 7.6 Hz, 1H),7.92-7.85 (m, 2H), 7.84 – 7.81 (m, 1H), 7.54 (dd, J = 8.9, 2.2 Hz, 1H), 6.69(t, J = 73.8Hz 1H), 6.61 (d, J = 8.4 Hz, 1H), 4.41-4.35 (m, 1H), 4.33-4.27(m, 1H), 4.00 (t, J = 5.5 Hz, 2H), 3.68 (s, 2H), 2.31-2.21 (m, 2H), 2.07 –1.94 (m, 2H), 1.65-1.55(m, 2H).

[0101] Example 9: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-1,2,3,4-tetrahydro-5 H -benzo[ eSynthesis of [1,4]diaza-5-one (a72)

[0102] The synthesis method was the same as in Example 1, and the product (19 mg) was obtained. ESI-MS (m / z): 482.20 [M+H] + . 1 H NMR (600 MHz, DMSO) -d6 ) δ 8.23 ​​(s, 2H), 7.87 (d, J = 2.6 Hz, 1H), 7.60 (dd, J =8.0, 1.5 Hz, 1H), 7.46 (d, J = 7.2 Hz, 1H), 7.30 (dd, J = 8.8, 2.6 Hz, 1H),7.21 – 7.16 (m, 1H), 7.03 (t, J = 74.4 Hz, 1H), 6.73 (d, J = 8.1 Hz, 1H), 6.65 (d, J = 6.5 Hz, 1H), 6.62 (dd, J = 11.4, 4.4 Hz, 1H), 6.48 (d, J = 8.8Hz, 1H), 6.36 (t, J = 3.7 Hz, 1H), 4.33 – 4.23 (m, 2H), 3.75 – 3.71 (m, 2H), 3.54 – 3.51 (m, 2H), 2.17-2.06 (m, 2H), 1.93 – 1.82 (m, 2H), 1.54 – 1.44 (m,2H).

[0103] Example 10: Synthesis of 6'-fluoro-1'-(6-(((1S,3S)-3-((5-(trifluoromethyl)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)spiro[cyclopropane-1,3'-indoline]-2'-one (compound a67)

[0104] The synthesis method was the same as in Example 1, and the product (12 mg) was obtained. ESI-MS (m / z): 499.18 [M+H] + . 1 H NMR (600 MHz, DMSO)-d6 ) δ 8.64 (s, 1H), 8.60 (s, 1H),8.21 (d, J = 7.3 Hz, 1H), 8.02 (s, 1H), 7.43 (dd, J = 8.8, 1.8 Hz, 1H), 7.11 (dd, J = 8.2, 5.4 Hz, 1H), 7.00 (d, J = 8.1 Hz, 1H), 6.88 – 6.82 (m, 1H), 6.62 (d, J = 8.1 Hz, 1H), 6.54 (dd, J = 9.3, 2.2 Hz, 1H), 4.49 – 4.41 (m, 1H), 4.36 (dd, J = 12.0, 5.9 Hz,1H), 2.22 – 2.10 (m, 2H), 2.01 – 1.88 (m, 2H), 1.70 (dd, J = 7.7, 3.8 Hz,2H), 1.61 – 1.59 (m, 2H), 1.58 – 1.49 (m, 2H).

[0105] Example 11: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[ f [1,4]oxynitric oxide-5(2 H Synthesis of )-ketone (compound a65)

[0106] The synthesis method was the same as in Example 1, and the product (14 mg) was obtained. ESI-MS (m / z): 483.20 [M+H] + .

[0107] Example 12: 4-(6-(((1S,3S)-3-((5-(trifluoromethyl)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[ f [1,4]Thiazine-5(2 H Synthesis of 1,1-ketodioxide (compound a86)

[0108] The synthesis method was the same as in Example 1, and the product (11 mg) was obtained. ESI-MS (m / z): 533.15 [M+H] + . 1 H NMR (600 MHz, DMSO) -d6 ) δ 8.64 (s, 1H), 8.59 (s, 1H), 8.20 (d, J = 7.2 Hz, 1H),8.06 (s, 1H), 7.96 (d, J = 7.6 Hz, 1H), 7.91 (t, J = 7.4 Hz, 1H), 7.84 (dd, J = 14.3, 7.5 Hz, 2H), 7.43 (d, J = 8.7 Hz, 1H), 6.82 (s, 1H), 6.55 (s, 1H), 4.43 (dd, J = 13.9, 6.9 Hz, 1H), 4.30 (d, J = 5.7 Hz, 1H), 3.90 (t, J = 5.5Hz, 2H), 3.75 (s, 2H), 2.23-2.09 (m, 2H), 1.99-1.83 (m, 2H), 1.63 – 1.47 (m, 2H).

[0109] Example 13: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydropyrido[4,3-] f [1,4]sulfur-nitrogen-5(2 H Preparation of 1,1-ketodioxide (compound a78)

[0110] The synthesis method was the same as in Example 1, and the product (20 mg) was prepared. ESI-MS (m / z): 532.15 [M+H] + . 1 H NMR (600 MHz, CD3OD) δ 9.14 (s, 1H), 9.09 (d, J = 2.5 Hz, 1H), 8.17 (s, 2H), 8.09(s, 1H), 7.84 (d, J = 4.8 Hz, 1H), 7.52 (dd,J = 8.9, 1.6 Hz, 1H), 6.69 (t, J = 72.0 Hz, 1H), 6.63 (s, 1H), 4.41 – 4.35 (m, 1H), 4.34 – 4.29 (m, 1H), 4.04(t, J = 5.8 Hz, 2H), 3.75 (t, J = 5.8 Hz, 2H), 2.31 – 2.21 (m, 2H), 2.05 –1.95 (m, 2H), 1.66 – 1.56 (m, 2H).

[0111] Example 14: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydropyrido[3,4-] f [1,4]sulfur-nitrogen-5(2 H Synthesis of 1,1-ketodioxide (compound a77)

[0112] The synthesis method was the same as in Example 1, and the product (18 mg) was obtained. ESI-MS (m / z): 532.15 [M+H] + . 1 H NMR (600 MHz, DMSO) -d6 ) δ 9.09 (d, J = 5.0 Hz, 1H), 9.01 (s, 1H), 8.23 ​​(s, 2H), 8.01 (d, J = 2.3 Hz, 1H), 7.90 (d, J = 5.0 Hz, 1H), 7.47 (d, J = 7.2 Hz, 1H), 7.41 (dd, J = 8.9, 2.5 Hz, 1H), 7.03 (t, J = 74.0 Hz, 1H), 6.82 (d, J = 6.8Hz, 1H), 6.53 (d, J = 8.9 Hz, 1H), 4.36-4.26 (m, 2H), 3.99 (t, J = 5.7 Hz, 2H), 3.85 (t, J= 5.4 Hz, 2H), 2.17 – 2.08 (m, 2H), 1.94 – 1.84 (m, 2H), 1.58– 1.46 (m, 2H).

[0113] Example 15: 4-(6-(((1S,3S)-3-((5-(difluoromethoxy)pyrimidin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydropyrido[2,3-] f [1,4]sulfur-nitrogen-5(2 H Synthesis of 1,1-ketodioxide (compound a76)

[0114] The synthesis method was the same as in Example 1, and the product (16 mg) was obtained. ESI-MS (m / z): 532.15 [M+H] + . 1 H NMR (600 MHz, CD3OD) δ 9.03 (s, 1H), 8.46 (d, J = 7.3 Hz, 1H), 8.18 (s, 2H), 7.87(s, 1H), 7.57 (d, J = 8.1 Hz, 1H), 6.69 (t, J = 78.0 Hz, 1H), 6.65 (s, 1H),4.60-4.56 (m, 1H), 4.42-4.30 (m, 2H), 4.11-4.05 (m, 2H), 3.78-3.70 (m, 2H),2.34 -2.21 (m, 2H), 2.09 – 1.92 (m, 2H), 1.66-1.58 (m, 2H).

[0115] Example 16: Synthesis of 4-(6-(((1S,3S)-3-((5-methylpyrazin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[f][1,4]thiazazide-5(2H)-one 1,1-dioxide (compound a104)

[0116] The synthesis method was the same as in Example 1, and the product (15 mg) was obtained. ESI-MS (m / z): 479.18 [M+H] + . 1 HNMR (600 MHz, Methanol- d4) δ 8.15 – 8.02 (m, 2H), 7.98 – 7.73 (m, 5H), 7.62 –7.47 (m, 1H), 6.66 – 6.57 (m, 1H), 4.41 – 4.23 (m, 2H), 4.08 – 3.94 (m, 2H), 3.69 (s, 2H), 2.40 – 2.19 (m, 5H), 2.06 – 1.89 (m, 2H), 1.69 – 1.54 (m, 2H).

[0117] Example 17: Synthesis of 4-(6-(((1S,3S)-3-((5-cyclopropylpyrazin-2-yl)amino)cyclopentyl)amino)pyridin-3-yl)-3,4-dihydrobenzo[f][1,4]thiazazide-5(2H)-one 1,1-dioxide (compound a109)

[0118] The synthesis method was the same as in Example 1, and the product (10 mg) was prepared. ESI-MS (m / z): 505.18 [M+H] + . 1 HNMR (600 MHz, Methanol- d 4) δ 8.12 – 8.04 (m, 2H), 7.94 – 7.80 (m, 4H), 7.76 (s, 1H), 7.54 (d, J = 8.8 Hz, 1H), 6.61 (d, J = 9.0 Hz, 1H), 4.30 (dt, J =17.7, 6.4 Hz, 2H), 4.05 – 3.96 (m, 2H), 3.69 (s, 2H), 2.33 – 2.19 (m, 2H), 2.02 – 1.88 (m, 3H), 1.65 – 1.54 (m, 2H), 0.95 – 0.85 (m, 4H).

[0119] Examples 18-109

[0120] Compounds of Examples 18-109 were synthesized using the synthetic method described in Example 1.

[0121] Comparative Example 1: (Compound 458B, namely AZD0780, in CN113574055B is synthesized using the method in this patent).

[0122] Experimental Example 1: In vitro LDL uptake assay of HepG2 cells

[0123] Table 1. Reagents and consumables used in in vitro LDL uptake experiments:

[0124] Experimental steps: 1. HepG2 cells were cultured in MEM medium with 10% FBS and a penicillin-streptomycin mixture of 100 units per milliliter, and cultured at 37°C under 5% CO2 conditions.

[0125] 2. Add 100 µL of cells at a density of 20,000 cells per well to a poly-D-lysine-coated 96-well culture plate. Incubate the cell culture plates overnight at 37°C and 5% CO2.

[0126] 3. Starve HepG2 cells using FBS-free MEM medium. Incubate the cell plates overnight at 37°C with 5% CO2.

[0127] 4. Starting from 40 mM, the test compound was diluted 3-fold in DMSO to obtain 7 dose concentrations.

[0128] 5. The diluted compound was diluted 100 times with MEM medium and then co-incubated with an equal volume of PCSK9 protein for 1 hour.

[0129] 6. Remove the supernatant from the cell plate and add 50 µL of the compound and protein mixture. Incubate the cell plate at 37°C and 5% CO2 for 23 h.

[0130] 7. Remove the cell culture medium from the plate and add 50 µL of BODIPY LDL dye. Incubate the cell culture plate at 37°C and 5% CO2 for 4 h.

[0131] 8. Remove the cell plate cleanser and wash 3 times with 100µL PBS.

[0132] 9. Calculate uptake by detecting fluorescence intensity at 480nm / 530nm using a high-content system.

[0133] Table 2. Effects of the compounds of this invention on restoring cellular LDL uptake activity

[0134] The experimental results show that the compounds of the present invention can significantly improve the LDL uptake capacity of HepG2 cells. Exemplary compounds are shown in Table 2.

[0135] Experimental Example 2: PCSK9 In Vitro Binding Assay

[0136] The ability of different compounds to bind to the PCSK9 protein was detected using the conventional surface plasmon resonance (SPR) method.

[0137] Test parameters

[0138] 1) Protein: PCSK9, Acro

[0139] 2) Chip: SA chip

[0140] 3) Experimental temperature: 25℃

[0141] 4) Experimental flow rate: 30 μL / min

[0142] 5) Fixing method: Tag capture method

[0143] 6) Combined measurement mode: multi-cycle dynamics

[0144] 7) Buffer solution:

[0145] a) Protein fixation: HBSP + 0.1 mM CaCl2, pH 7.4

[0146] b) Compound analysis: HBSP + 0.1 mM CaCl2, pH 7.4, 4% DMSO

[0147] protein fixation

[0148] 1) The PCSK9 (Acro) protein is immobilized onto the SA chip via biotin-SA tag capture, which closes the channel after immobilization.

[0149] 2) The reference channel is closed.

[0150] Analysis and Testing

[0151] 1) Prepare compound analysis buffer (HBSP + 0.1 mM CaCl2, pH 7.4, 4% DMSO) and filter it through a 0.22 μm filter membrane.

[0152] 2) Prepare a series of compounds with different concentration gradients, a total of 8 concentrations, and perform multiple cycles of detection.

[0153] Data Analysis

[0154] The final binding and dissociation curves were obtained by subtracting the reference channel and buffer blank control from the experimental channel signal values. The affinity data were obtained by fitting the curves using the 1:1 binding mode kinetics method.

[0155] Experimental results show that the compounds of this application have a high affinity for PCSK9 protein, and exemplary compounds are shown below.

[0156] Table 3. Binding activity of the compounds of the present invention against PCSK9 protein

[0157] A: Less than 10 nM; B: Greater than or equal to 10 nM and less than 100 nM.

[0158] Experimental Example 3: Mouse PK Test

[0159] ICR mice (Beijing Vitonda Biotechnology Co., Ltd.) were administered different compounds by gavage (5 mg / kg) or intravenously (1 mg / kg). At different time points after administration (gavage group: 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h; intravenous group: 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 24 h), 100 μL of blood was collected from the orbital sinus of the mice and placed in a heparin sodium anticoagulant tube. Plasma samples were obtained by centrifugation at 3000 rpm for 10 min within 2 h. Methanol protein precipitation was used, and the drug concentration in the mouse plasma after administration was determined by HPLC-MS / MS. Drug-time curves were plotted, and pharmacokinetic parameters were calculated. The pharmacokinetic behavior of the compounds in mice after administration was described by statistical moment parameters of a non-compartmental model.

[0160] Table 4. Pharmacokinetic parameters of mice administered via gavage at 5 mg / kg

[0161] The experimental results show that the compound of this application has good in vivo pharmacokinetics and has the potential to become a drug.

[0162] Experimental Example 4: In vivo efficacy test of B6-hPCSK9-CDS mouse model

[0163] B6-hPCSK9-CDS transgenic C57 mice (5-6 weeks old, male) were introduced to the barrier system and allowed to acclimatize for 3-5 days before being fed Western Diets (Changzhou Shuyi Shuer Biotechnology Co., Ltd., catalog number 12079B). After 5-6 weeks of feeding, blood was collected from the orbital sinus of the mice into EP tubes. After standing at room temperature for 30 minutes, the samples were centrifuged at 5000 rpm for 10 minutes to collect serum samples. The serum LDL-c level was measured using a fully automated blood biochemistry analyzer. The serum LDL-c level was measured again after 1 week. Once the level stabilized, the mice were randomly assigned to different groups based on their LDL-c levels and given appropriate drug treatments. The indicators monitored during the experiment were body weight and serum LDL-c level.

[0164] The experimental results show that the compound of this application can effectively reduce the level of LDL-c in serum and has the potential to become a drug.

[0165] Experiment Example 5: In vivo PK test in cynomolgus monkeys

[0166] Cynomolgus monkeys (Guangxi Guidong Primate Development and Experiment Co., Ltd.) were administered a single compound via gavage to investigate the pharmacokinetic (PK) behavior of different compounds. The compound was administered at a dose of 1.5 mg / kg. At different time points after administration (0 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, and 48 h), 0.5 mL of whole blood was collected from the forelimb vein and placed in EP tubes containing EDTA-K2. Plasma samples were obtained by centrifugation at 2000 rpm for 10 min within 30 min. Methanol protein precipitation was used, and the drug concentration in the cynomolgus monkey plasma was determined by HPLC-MS / MS. Drug-time curves were plotted, and pharmacokinetic parameters were calculated. The pharmacokinetic behavior of the compounds in cynomolgus monkeys after administration was described using non-compartmental model statistical moment parameters.

[0167] Table 5. Pharmacokinetic parameters of monkeys administered 1.5 mg / kg via gavage

[0168] The experimental results show that the compound of this application has good in vivo pharmacokinetics and has the potential to become a drug.

[0169] Experimental Example 6: In vitro inhibition of human potassium ion channels (hERG)

[0170] Experimental equipment: Electrophysiological detection was performed using a fully automated patch clamp QPatch 48 X (Sophion) device.

[0171] Experimental Procedure: Prepared cells were placed on a centrifuge in a Qpatch workbench and washed using multiple centrifugation / resuspension methods to replace the cell culture medium with extracellular fluid. An MTP-96 plate was removed and placed in the MTP source position. The QPlate chip was removed and placed in the Qplate source position. A robotic arm scanned the barcodes on the MTP-96 plate and the QPlate chip and picked them up to the measurement station. Intracellular and extracellular fluids were aspirated from the liquid pools and added to the intracellular fluid pool and cell and test substance pool of the QPlate chip, respectively. At the measurement station, all measurement sites on the QPlate underwent initial quality control. The quality control process included aspirating cell suspension from the centrifuge's cell container and positioning the cells onto the chip wells using a pressure controller to establish a high-resistance seal, forming a whole-cell recording mode. Once a stable control current baseline was obtained, the test substance was aspirated from the MTP-96 plate according to the concentration gradient and applied to the cells. The current detected in the compound-free extracellular fluid for each cell served as its control group, and two cells were independently tested repeatedly. All Qpatch electrophysiological experiments were performed at 24°C.

[0172] Compound treatment: The compound was diluted in gradients of 0.3, 1, 3, 10, and 30 μM.

[0173] Data Processing: Computing IC 50 value.

[0174] The experimental results show that the compound of the present invention does not have a significant inhibitory effect on the hERG channel within the detection concentration range of this experiment, indicating that the compound of the present invention has a low risk of cardiotoxicity.

Claims

1. A compound of formula (I), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound: （I） Ring A is selected from 5-10 member heterocyclic groups, C 6-10 Aryl or 5-10 heteroaryl groups; R a Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, -C(O)NR1R2, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 cycloalkyl, phenyl, the C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 The cycloalkyl or phenyl group is optionally surrounded by one, two, or three groups selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-C. 1-4 Alkyl, -C(O)-C 1-4 Substituents of alkyl groups; R1 and R2 are independently selected from hydrogen, deuterium, and C, respectively. 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 Cycloalkyl, 4-6 membered heterocyclic groups, halogenated C 1-6 Alkyl, hydroxyl substituted C 1-6 alkyl; Ring B is selected from 9-16 membered bicyclic or tricyclic heterocyclic groups; R b Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, oxo, thio, =NH, =NC respectively. 1-4 Alkyl, =NC 3-6 cycloalkyl, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 Cycloalkyl, 5-6 membered heterocyclic, phenyl, 5-6 membered heteroaryl, wherein C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 1-6 Alkylthio, C 3-6 The cycloalkyl, 5-6 membered heterocyclic, aryl, and 5-6 membered heteroaryl groups are optionally substituted by one, two, or three substituents selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, and cyano groups; or two R groups attached to the same atom. b The linkage forms a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally surrounded by 1, 2 or 3 groups selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl thiols, halogenated C 1-4 Alkyl, Halogenated C 1-4 Alkoxy, halogenated C 1-4 The substituents of the alkylthio group are replaced; R c Selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkyl thiols, halogenated C 1-4 Alkyl, Halogenated C 1-4 Alkoxy, halogenated C 1-4 Alkylthio; x is 0, 1, 2, 3 or 4; y is 0, 1, 2, 3 or 4; p can be 0, 1, 2, 3, or 4.

2. The compound of claim 1, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Ring A is selected from C 6-10 The ring is aryl or 5-10-membered heteroaryl; more preferably, ring A is selected from phenyl, 5-membered monocyclic heteroaryl, 6-membered monocyclic heteroaryl, 5-membered 6-membered bicyclic heteroaryl, 6-membered 5-membered bicyclic heteroaryl, 5-membered 5-membered bicyclic heteroaryl, 6-membered 6-membered bicyclic heteroaryl; even more preferably, ring A is selected from 5-membered monocyclic heteroaryl, 6-membered monocyclic heteroaryl, 5-membered 6-membered bicyclic heteroaryl, 6-membered 5-membered bicyclic heteroaryl; Preferably, ring A is selected from , , , , , , . , , ; More preferably, ring A is selected from .

3. The compound of claim 1 or 2, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, R a Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, -C(O)NR1R2, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 cycloalkyl, phenyl, the C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 The cycloalkyl or phenyl group is optionally surrounded by one, two, or three groups selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-C. 1-3 Alkyl, -C(O)-C 1-3 Substituents of alkyl groups; R1 and R2 are independently selected from hydrogen, deuterium, and C, respectively. 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 Cycloalkyl, 4-6 membered heterocyclic groups, halogenated C 1-3 Alkyl, hydroxyl substituted C 1-3 alkyl; Preferred, R a The groups are selected from hydrogen, deuterium, halogen, cyano, -C(O)NR1R2, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylthio, ethylthio, n-propylthio, isopropylthio, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and phenyl, respectively. The methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, n-propoxy, isopropoxy, methylthio, ethylthio, n-propylthio, isopropylthio, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and phenyl groups are optionally substituted with 1, 2, or 3 substituents selected from deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, -SO2-CH3, and -C(O)-CH3. R1 and R2 are each independently selected from hydrogen, deuterium, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, oxacyclopentyl, halomethyl, haloethyl, halon-propyl, haloisopropyl, hydroxy-substituted methyl, hydroxy-substituted ethyl, hydroxy-substituted n-propyl, and hydroxy-substituted isopropyl; more preferably, one of R1 and R2 is selected from hydrogen, deuterium, methyl, and ethyl, and the other is selected from methyl, ethyl, n-propyl, isopropyl, cyclopropyl, oxacyclopentyl, halomethyl, haloethyl, halon-propyl, haloisopropyl, hydroxy-substituted methyl, hydroxy-substituted ethyl, hydroxy-substituted n-propyl, and hydroxy-substituted isopropyl. More preferably, R a The groups selected are hydrogen, F, Cl, Br, methyl, ethyl, methoxy, methylthio, -OCHF2, -OCH2F, -OCF3, monofluoromethyl, difluoromethyl, trifluoromethyl, cyclopropyl, cyclobutyl, hydroxymethyl, hydroxyethyl, hydroxy-substituted cyclopropyl, hydroxy-substituted cyclobutyl, cyano, -CH2-SO2-CH3, -CH2-CO-CH3, phenyl, -CONH2, -CONHCH3, -CON(CH3)CH2CH2OH, respectively. Even more preferred, R a It is -OCHF2, trifluoromethyl.

4. The compound according to any one of claims 1-3, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Ring B is selected from 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 5-membered and 7-membered bicyclic heterocyclic groups, 7-membered and 5-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 7-membered bicyclic heterocyclic groups, 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 5-membered and 7-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 7-membered bicyclic heterocyclic groups; more preferably, ring B is selected from 5-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 5-membered bicyclic heterocyclic groups, 6-membered and 6-membered bicyclic heterocyclic groups, 7-membered and 6-membered bicyclic heterocyclic groups, 6-membered and 7-membered bicyclic heterocyclic groups; Preferably, ring B is selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

5. The compound according to any one of claims 1-4, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, R b Selected from hydrogen, deuterium, halogen, amino, hydroxyl, cyano, nitro, oxo, thio, =NH, =NC respectively. 1-3 Alkyl, =NC 3-6 cycloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-6 cycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Alkylthio, C 3-6 The cycloalkyl group may be optionally substituted by one, two, or three substituents selected from deuterium, halogen, nitro, amino, hydroxyl, and cyano; or two R groups attached to the same atom. b The linkage forms a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally surrounded by 1, 2 or 3 groups selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 The substituents of the alkylthio group are replaced; Preferred, R b The radicals are selected from hydrogen, F, Cl, Br, amino, hydroxyl, cyano, oxo, thio, =NH, =N-CH3, =N-cyclopropane, =N-cyclobutane, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, methylthio, ethylthio, and cyclopropyl, wherein the methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, methylthio, ethylthio, cyclopropyl, and cyclobutane radicals are optionally substituted by 1, 2, or 3 substituents selected from halogens and cyano groups; or two R radicals attached to the same atom. b The links form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, azirne, azirne, cyclohexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, azirne, azirne are optionally substituted by 1, 2 or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, ethyl, methoxy, ethoxy; More preferably, R b The radicals are selected from hydrogen, F, Cl, Br, cyano, oxo, thio, =NH, =N-CH3, =NH-cyclopropane, =N-cyclobutane, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, cyclopropyl; or two R radicals attached to the same atom. b The links form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, azirne, azirne, cyclohexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, azirne, azirne are optionally substituted by 1, 2 or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, methoxy.

6. The compound according to any one of claims 1-5, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

7. The compound according to any one of claims 1-6, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, R c Selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; more preferably, R c Selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy; further preferably, R c Selected from hydrogen.

8. The compound according to any one of claims 1-7, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, x is 0, 1, 2 or 3; preferably, x is 0, 1 or 2.

9. The compound according to any one of claims 1-8, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, y can be 0, 1, 2 or 3.

10. The compound according to any one of claims 1-9, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, p is 0 or 1.

11. The compound shown in any one of claims 1-10, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, further as shown in formula (II): (II) in, Each E is independently selected from CR d Or N, R d Independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; R b1 and R b2 The C atoms attached thereto form a 3-6 membered cycloalkyl group or a 4-6 membered heterocyclic group, wherein the 3-6 membered cycloalkyl group or the 4-6 membered heterocyclic group is optionally composed of 1, 2 or 3 atoms selected from hydrogen, deuterium, halogen, nitro, amino, hydroxyl, oxo, thio, carboxyl, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 The substituents of the alkylthio group are replaced; Rings A and R a R c X and p are as described in equation (I).

12. The compound of claim 11, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Each E is a CR d Or, only one E is N, and the others are CR. d R d Independently selected from hydrogen, deuterium, halogen, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; more preferably, R d Independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; further preferably, R d Independently selected from hydrogen, deuterium, F, Cl, Br, cyano, methyl, and methoxy; further preferably, R d It is independently selected from hydrogen, F, Cl, and Br.

13. The compound of claim 11 or 12, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, R b1 and R b2 The C atoms connected thereto together form cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, oxetane, oxetane, azirne, azirne, and azirnehexane, wherein the cyclopropane, cyclobutane, cyclopentane, cyclohexane, oxetane, oxetane, azirne, azirne, and azirnehexane are optionally substituted by 1, 2, or 3 substituents selected from hydrogen, F, Cl, Br, cyano, methyl, ethyl, methoxy, and ethoxy. Preferred, R b1 and R b2 The C atoms connected thereto form cyclopropane, cyclobutane, oxecyclobutane, oxecyclopentane, oxecyclohexane, and azeocyclobutane, wherein the cyclopropane, cyclobutane, oxecyclobutane, oxecyclopentane, oxecyclohexane, and azeocyclobutane are optionally substituted by 1, 2, or 3 substituents selected from hydrogen, F, Cl, Br, and methyl.

14. The compound of claim 11, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Selected from or R b1 and R b2 The C atoms attached thereto form cyclopropane and aziridine, wherein the cyclopropane and aziridine are optionally substituted by one, two, or three substituents selected from hydrogen, F, Cl, Br, and methyl; R d It is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, and trifluoromethyl.

15. The compound shown in any one of claims 1-10, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, further as shown in formula (III): (III) in, Each G is independently selected from CR g Or N, R g Independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkyl thiols, halogenated C 1-3 Alkyl, Halogenated C 1-3 Alkoxy, halogenated C 1-3 Alkylthio; K is selected from -S-, -O-, -S(O)-, -S(O)2-, -C(R) k ) (R k )-、-S(O)(=NR k - or -N(R) k )-, where each R k Each element is independently selected from hydrogen, deuterium, halogen, cyano, and C. 1-3 Alkyl, C 1-3 Alkoxy, C 1-3 Alkylthio, C 3-6 cycloalkyl; Rings A and R a R b R c X, y, and p are as described in equation (I).

16. The compound of claim 15, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, Each G is a CR g Or, only one G is N, and all others G are CR. g R g Independently selected from hydrogen, deuterium, halogen, cyano, methyl, ethyl, methoxy, ethoxy, methylthio, ethylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; more preferably, R g Independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, trifluoromethyl; further preferably, R g Independently selected from hydrogen, deuterium, F, Cl, Br, cyano, methyl, and methoxy; further preferably, R g It is independently selected from hydrogen.

17. The compound of claim 15 or 16, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, K is selected from -S-, -O-, -S(O)-, -S(O)2-, -C(R) k (R) k )-、-S(O)(NR k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, cyclopropyl, and cyclobutyl; preferably, K is selected from -S-, -O-, -S(O)-, -S(O)2-, and -C(R). k ) (R k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, halogen, cyano, methyl, and methoxy; more preferably, K is selected from -S-, -O-, -S(O)2-, and -C(R). k ) (R k - or -N(R) k )-, where each R k Each of the following is independently selected from hydrogen, deuterium, F, Cl, Br, and methyl; more preferably, K is selected from -S-, -O-, -S(O)2-, -CH2-, or -NH-.

18. The compound of claim 15, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, wherein, K is selected from -S-, -O-, -S(O)2- or -NH-, and G is always CR. g Or, only one G is N, and all others G are CR. g R g It is independently selected from hydrogen, deuterium, halogen, cyano, methyl, methoxy, methylthio, monofluoromethyl, difluoromethyl, and trifluoromethyl.

19. A compound, characterized in that, It has the following structure: Or its stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compound.

20. A compound, characterized in that, It has the following structure: Or its stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compound.

21. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a compound as shown in any one of claims 1-20, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound; further, the pharmaceutical composition also comprises a pharmaceutically acceptable excipient.

22. Use of the compound of any one of claims 1-20, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 21, in the preparation of a medicament for treating and / or preventing PCSK9-mediated diseases; preferably, PCSK9-mediated diseases are selected from cardiovascular diseases, liver diseases, infectious and autoimmune diseases, neurocognitive disorders, and cancer.

23. The use of the compound of any one of claims 1-20, a stereoisomer, tautomer or mixture thereof, or a pharmaceutically acceptable salt of the compound, or the pharmaceutical composition of claim 21 in the preparation of a medicament for treating and / or preventing LDL reduction.

24. Use of the compound shown in any one of claims 1-20, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound, or the pharmaceutical composition of claim 21, in the preparation of a medicament for the treatment and / or prevention of cardiovascular diseases, liver diseases, infectious and autoimmune diseases, neurocognitive disorders, and cancer.

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