PCSK9 cyclic peptide inhibitor, preparation method therefor, and use thereof

Abstract

A PCSK9 cyclic peptide inhibitor or a stereoisomer, a racemate, or a pharmaceutically acceptable salt thereof, a preparation method therefor, and a use thereof. Compound (I) disclosed herein has good PCSK9 inhibitory activity and can be used for treating diseases such as hyperlipidemia.

Description

A PCSK9 cyclic peptide inhibitor, its preparation method and uses Technical Field

[0001] This invention belongs to the field of chemical pharmaceutical technology, specifically relating to a PCSK9 cyclic peptide inhibitor, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, as well as its preparation method and uses. The compound provided by this invention has excellent PCSK9 inhibitory activity and can be used to treat diseases such as hyperlipidemia. Background Technology

[0002] Proprotein convertase subtilisin / kexin type 9 (PCSK9), also known as neuronal apoptosis-regulating convertase 1 (NARC-1), is a pro-proprotein convertase belonging to the subtilisin (S8) family of serine proteases. It is expressed in cells capable of proliferation and differentiation, including hepatocytes, renal interstitial cells, ileal and colonic epithelial cells, and embryonic telencephalon neurons. Studies have found that PCSK9 plays a role in the differentiation of hepatocytes and nerve cells. It not only specifically acts on cholesterol biosynthesis or uptake, but circulating PCSK9 can also directly bind to the low-density lipoprotein receptor (LDLR) on the surface of hepatocytes, being phagocytosed by hepatocytes along with LDLR. This promotes the degradation of LDLR in hepatocytes, hinders its recycling, and thus increases the level of LDL cholesterol (LDL-C) in plasma. Elevated LDL-C expression is closely related to dyslipidemia and cardiovascular diseases in humans. Summary of the Invention

[0003] In view of the problems existing in the prior art, the present invention provides a PCSK9 cyclic peptide inhibitor, or its stereoisomer, racemate or pharmaceutically acceptable salt thereof, as well as its preparation method and uses. The compound provided by the present invention has good PCSK9 inhibitory activity and can be used to treat diseases such as hyperlipidemia.

[0004] This invention is achieved through the following technical solution:

[0005] This invention provides a PCSK9 cyclic peptide inhibitor, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, the structure of which is shown in general formula (I):

[0006] in,

[0007] R1 is selected from substituted or unsubstituted phenyl, 5-10-membered heteroaryl, and the substituent is selected from halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkyl, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, C3-C6 cycloalkylalkyl, C3-C6 cycloalkyloxy, C3-C6 heterocycloalkyloxy, amino, alkyl-substituted amino, aminoalkyl, aminoalkoxy, alkyl-substituted aminoalkyl, or two substituents forming cycloalkyl or heterocycloalkyl;

[0008] R2, R 3a R 3b R4, R5, R6, R7, and R8 are all independently selected from H, halogens, C1-C6 alkyl groups, halo-C1-C6 alkyl groups, or R7 and R8 are cycloalkyl groups;

[0009] Y is selected from hydroxyl or amino groups;

[0010] T1, T2, and T3 are all independently selected from substituted or unsubstituted C1-C6 alkyl, C1-C6 alkyl-benzene-C1-C6 alkyl, C1-C6 alkyl-5-10 heteroaryl-C1-C6 alkyl, and the substituents are selected from halogen, C1-C6 alkyl, and C1-C6 alkoxy.

[0011] Connect T1 to C or T2;

[0012] T4 is selected from substituted or unsubstituted C1-C6 alkyl groups, and the substituents are selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, halo-C1-C6 alkyl groups, halo-C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, C3-C6 heterocyclic alkyl groups, C3-C6 cycloalkylalkyl groups, C3-C6 cycloalkyloxy groups, C3-C6 heterocyclic alkyloxy groups, amino groups, alkyl-substituted amino groups, aminoalkyl groups, alkyl-substituted aminoalkyl groups, or two substituents forming cycloalkyl or heterocyclic alkyl groups;

[0013] Ring A is selected from substituted or unsubstituted phenyl groups, 5-10-membered heteroaryl groups, and the substituents are selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, and halo-C1-C6 alkyl groups.

[0014] B is selected from -C(O)-NH- or -NH-C(O)-, or B forms a heterocyclic alkyl group with T4, and the heterocyclic alkyl group can be further replaced by hydroxyl, halogen, alkyl, or haloalkyl.

[0015] C is selected from -C(O)-N- or N;

[0016] D is selected from Among them, X1, X2, X3, and X4 are all independently selected from CH or N. When it is CH, it can be further substituted by halogens or C1-C6 alkyl groups.

[0017] X5, X6, X7, and X8 are all independently selected from C, CH, or N. There is a single or double bond between X5 and X6, and a single or double bond between X7 and X8. Alternatively, one of X6 or X7 may not exist. When it is CH, it may be further substituted by halogens or C1-C6 alkyl groups.

[0018] R9 is selected from alkyl or alkylamino groups, and the alkyl or alkylamino groups may be further replaced by halogens or C1-C6 alkyl groups;

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

[0020] R 10 R 11 Each is independently selected from H, halogen, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or R. 10 R 11 Formation of cycloalkyl groups;

[0021] R 12 Selected from C1-C6 alkyl groups or not present;

[0022] Wherein, R2 is not hydrogen;

[0023] And / or when T4 is CH3-CH, B is not -NH-C(O)-;

[0024] And / or when R 3a Methyl, R 3b When Y is hydrogen, it is not a hydroxyl group;

[0025] and / or R1 is not

[0026] And / or when m is 2, R 10 It is not methyl.

[0027] Furthermore, as a preferred embodiment of the present invention, the structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (II):

[0028] Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X6, and m are the same as above.

[0029] Furthermore, as a preferred embodiment of the present invention, the structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (III):

[0030] Among them, R1-R12 The definitions of Y, T1-T4, AC, X1, X3, X7, and m are the same as in claim 1.

[0031] Furthermore, as a preferred embodiment of the present invention, the structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (IV):

[0032] Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X6, and m are the same as above.

[0033] Furthermore, as a preferred embodiment of the present invention, the structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (V):

[0034] Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X7, X8, and m are the same as above.

[0035] Furthermore, as a preferred embodiment of the present invention, the halogen is preferably F, Cl, Br, or I.

[0036] Further, as a preferred embodiment of the present invention, the alkyl group is preferably C1-C6, and more preferably C1-C6 alkyl groups are C1-C2, C1-C3, C1-C4, or C1-C5 alkyl groups; examples of the alkyl group include: methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, and 1-ethylbutyl.

[0037] Furthermore, as a preferred embodiment of the present invention, the cycloalkyl group is preferably a spirocyclic, bridged, fused ring, or C3-C6 cycloalkyl group; the C3-C6 cycloalkyl group is preferably cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; and the spirocyclic group is preferably... Bridge ring selection: Fused ring preferred: Heterocyclic alkyl groups are cycloalkyl groups in which one or more carbon atoms are replaced by heteroatoms, which are selected from oxygen, sulfur, and NH.

[0038] Furthermore, as a preferred embodiment of the present invention, the C 1-C6 haloalkyl groups are preferably C1-C2, C1-C3, C1-C4, or C1-C5 haloalkyl groups; examples of such haloalkyl groups include: -CHF2, -CHCl2, -CF3, -CCl3, -CHFCH2F, -CHClCH2F, -CF2CHF2, -CH2CHF2, -CH2CF3, -CHFCH3, -CH2CH2F, -CF2CH3, -CH2CF2CHF2, etc., and in one embodiment, C 1- C6 haloalkyl groups include C6 groups with fluorine substitution. 1- C6 alkyl; in another embodiment, C 1- C4 haloalkyl groups include C4 groups with fluorine substitution. 1- C4 alkyl; in another embodiment, C 1- C3 haloalkyl groups include C3 fluorinated alkyl groups. 1- C3 alkyl.

[0039] Furthermore, as a preferred embodiment of the present invention, the C1-C6 alkoxy group is preferably C1-C2, C1-C3, C1-C4, or C1-C5 alkoxy group. More specifically, the alkoxy group is selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0040] Furthermore, as a preferred embodiment of the present invention, C 1- The preferred C6 haloalkoxy group is C1-C2, C1-C3, C1-C4, or C1-C5 haloalkoxy group. Further, the haloalkoxy group is specifically selected from -OCHF2, -OCHCl2, -OCF3, -OCCl3, -OCHFCH2F, -OCHClCH2F, -OCF2CHF2, -OCH2CHF2, -OCH2CF3, -OCHFCH3, -OCH2CH2F, -OCF2CH3, -OCH2CF2CHF2, etc.

[0041] Furthermore, as a preferred embodiment of the present invention, the 5-10-membered heteroaryl group is selected from: 5-8-membered heteroaryl groups and 5-6-membered heteroaryl groups. Examples of the heteroaryl group include, but are not limited to: pyrrole, furanyl, thiophene, imidazolyl, furazonyl, oxazolyl, oxadiazolyl, oxtriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, tetrazolyl, pyridyl, pyrazinyl, pyridinyl, pyrimidinyl, and triazinyl.

[0042] Furthermore, as a preferred embodiment of the present invention, A is a phenyl group;

[0043] Furthermore, as a preferred embodiment of the present invention, T1 and / or T3 are alkyl groups, and / or T2 is...

[0044] Furthermore, as a preferred technical solution of the present invention, The definitions of T4 and B are the same as above. Further, as a preferred embodiment of the present invention, T4 is selected from substituted or unsubstituted C1-C6 alkyl groups, and the substituents are selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, halogenated C1-C6 alkyl groups, halogenated C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, C3-C6 heterocyclic alkyl groups, C3-C6 heterocyclic alkyl groups, C3-C6 cycloalkyloxy groups, C3-C6 heterocyclic alkyloxy groups, amino groups, alkyl-substituted amino groups, aminoalkyl groups, alkyl-substituted aminoalkyl groups, or two substituents forming a cycloalkyl or heterocyclic alkyl group; B is selected from -C(O)-NH-, or -NH-C(O)-, or B and T4 forming a heterocyclic alkyl group, which may be further substituted by hydroxyl groups, halogens, alkyl groups, or halogenated alkyl groups.

[0045] Furthermore, as a preferred embodiment of the present invention, *-T4-B-* is selected from...

[0046] Furthermore, as a preferred embodiment of the present invention, D is...

[0047] Furthermore, as a preferred embodiment of the present invention, R1 is...

[0048] Furthermore, as a preferred embodiment of the present invention, R8 is H or methyl.

[0049] Furthermore, as a preferred embodiment of the present invention, R9 is... or Furthermore, R9 is or

[0050] Where * indicates a connection position. + It represents a cation that forms a salt with a suitable acid.

[0051] Furthermore, the present invention also provides a PCSK9 cyclic peptide inhibitor, or its stereoisomer, its racemic mixture, or its pharmaceutically acceptable salt, characterized in that the PCSK9 cyclic peptide inhibitor is selected from: A1-A7, B1-B12, C1-C3, D1-D29, E1-E8, F1-F2, G1-G4.

[0052] Furthermore, the present invention also provides a pharmaceutical composition characterized in that it comprises a PCSK9 cyclic peptide inhibitor of formula (I) or formula (II), or a stereoisomer thereof, or a racemic mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients and / or carriers.

[0053] Furthermore, the present invention also provides the use of a pharmaceutical composition comprising a PCSK9 cyclic peptide inhibitor of formula (I), formula (II), formula (III), formula (IV), or formula (V), or a stereoisomer thereof, or a racemic mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a PCSK9 cyclic peptide inhibitor of formula (I), formula (II), formula (III), formula (IV), or formula (V) in the preparation of a medicament for the prevention or treatment of diseases related to the PCSK9 receptor.

[0054] Furthermore, as a preferred embodiment of the present invention, the PCSK9 receptor-related disease is selected from: hyperlipidemia.

[0055] Furthermore, the present invention also provides a method for preparing a PCSK9 cyclic peptide inhibitor, or its stereoisomer, its racemic mixture, or its pharmaceutically acceptable salt, which can be prepared using conventional methods in the art, or by referring to the method of patent CN112313243A and methods known in the art.

[0056] In the chemical structure of the compound described in this invention, the bond... This indicates that the configuration is not specified; that is, if chiral isomers exist in the chemical structure, the bond... It can be Or simultaneously include Two configurations;

[0057] In the chemical structure of the compounds described in this disclosure, the bonds... The configuration is not specified, meaning it can be either Z configuration or E configuration, or both configurations can be included simultaneously;

[0058] The compounds and intermediates of the present invention may also exist in different tautomer forms, and all such forms are included within the scope of this disclosure. The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert via low energy barriers. For example, proton tautomers (also called proton transfer tautomers) include interconversions via proton transfer, such as keto-enol and imine-enamine, lactam-lactamimide isomerization. Examples of lactam-lactamimide equilibrium are between A and B as shown below.

[0059] All compounds in this invention may be designated as type A or type B. All tautomers are within the scope of this disclosure. The nomenclature of compounds does not exclude any tautomers.

[0060] For clarity, this article defines the general terminology used in the description of compounds.

[0061] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without undue toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0062] The term "alkyl" refers to a branched, unbranched, or cyclic saturated hydrocarbon chain containing a specified number of carbon atoms. The alkyl group is preferably C16-264-3 ... 1- C6 alkyl, the C 1- The number of carbon atoms in a C6 alkyl group is selected from 1, 2, 3, 4, 5, or 6, wherein the C... 1- C6 alkyl groups are preferably C1-C2, C1-C3, C1-C4, or C1-C5 alkyl groups; examples of such alkyl groups include: methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, and 1-ethylbutyl.

[0063] The term "halogenated alkyl" means that the hydrogen atom on the alkyl group can be replaced by one or more halogen atoms, preferably C16. 1- C6 haloalkane, wherein the C 1- The number of carbon atoms in the C6 haloalkyl group is selected from 1, 2, 3, 4, 5, or 6, wherein the C... 1- C6 haloalkyl groups are preferably C1-C2, C1-C3, C1-C4, or C1-C5 haloalkyl groups; examples of such haloalkyl groups include: -CHF2, -CHCl2, -CF3, -CCl3, -CHFCH2F, -CHClCH2F, -CF2CHF2, -CH2CHF2, -CH2CF3, -CHFCH3, -CH2CH2F, -CF2CH3, -CH2CF2CHF2, etc., and in one embodiment, C 1-C6 haloalkyl groups include C6 groups with fluorine substitution. 1- C6 alkyl; in another embodiment, C 1- C4 haloalkyl groups include C4 groups with fluorine substitution. 1- C4 alkyl; in another embodiment, C 1- C3 haloalkyl groups include C3 fluorinated alkyl groups. 1- C3 alkyl.

[0064] The term "alkoxy" refers to an alkyl group in which one or more carbon atoms are replaced by oxygen, such as -O-(alkyl), wherein the definition of alkyl is as described above. The number of carbon atoms in the C1-C6 alkoxy group is selected from 1, 2, 3, 4, 5 or 6, and the alkoxy group is preferably C1-C6 alkoxy. The C1-C6 alkoxy group is preferably C1-C2, C1-C3, C1-C4 or C1-C5 alkoxy. Further, the alkoxy group is specifically selected from methoxy, ethoxy, propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, and tert-butoxy.

[0065] The term "haloalkoxy" refers to an alkoxy group in which one or more hydrogen atoms are substituted by halogens; the haloalkoxy group is preferably C64-C ... 1- C6 haloalkoxy groups, wherein the number of carbon atoms in the C1-C6 haloalkoxy groups is selected from 1, 2, 3, 4, 6, or 5, and the C1-C6 haloalkoxy groups are preferably C1-C2, C1-C3, C1-C4, or C1-C5 haloalkoxy groups. Further, the haloalkoxy groups are specifically selected from -OCHF2, -OCHCl1, -OCF3, -OCCl3, -OCHFCH1F, -OCHClCH1F, -OCF1CHF1, -OCH2CHF1, -OCH1CF3, -OCHFCH3, -OCH2CH2F, -OCF2CH3, -OCH2CF2CHF2, etc. In one embodiment, C... 1- C6 haloalkoxy groups include C with fluorine substitution. 1- C6 alkoxy; in another embodiment, C 1- C4 haloalkyl groups include C4 groups with fluorine substitution. 1- C4 alkoxy; in yet another embodiment, C 1- C3 haloalkyl groups include C3 fluorinated alkyl groups. 1- C3 alkoxy group.

[0066] The term "heteroaryl" refers to a monocyclic or fused polycyclic aromatic ring structure comprising one or more (preferably 1, 2, 3, or 4) heteroatoms independently selected from O, N, and S, and a specified number of carbon atoms. Specifically, the aromatic ring structure may have 5 to 12 ring members. The heteroaryl is preferably a 5- to 10-membered ring, more preferably a 5- to 8-membered ring, and most preferably a five-, six-, or seven-membered ring. The heteroaryl can be, for example, a five- or six-membered monocyclic ring or a fused bicyclic structure formed by fused five- and six-membered rings or two fused six-membered rings or, as another example, two fused five-membered rings. Each ring may contain up to four heteroatoms, typically selected from nitrogen, sulfur, and oxygen. The heteroaryl ring typically contains up to four heteroatoms, more typically up to three heteroatoms, and more typically up to two heteroatoms, such as a single heteroatom. In one embodiment, the heteroaryl ring contains at least one cyclic nitrogen atom. The nitrogen atom in a heteroaryl ring can be basic, as in the case of imidazole or pyridine, or substantially non-basic, as in the case of indole or pyrrole nitrogen. Generally, the number of basic nitrogen atoms present in a heteroaryl ring (including any amino substituents in the ring) will be less than five.

[0067] Examples of five-membered monocyclic heteroaryl groups include (but are not limited to) pyrrole, furanyl, thiophene, imidazolyl, furazonyl, oxazolyl, oxadiazolyl, oxtriazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, triazolyl, and tetrazolyl. Examples of six-membered monocyclic heteroaryl groups include (but are not limited to) pyridyl, pyrazinyl, pyridazinyl, pyrimidinyl, and triazinyl. Specific examples of bicyclic heteroaryl groups containing a five-membered ring fused to another five-membered ring include (but are not limited to) imidazothiazolyl and imidazothiazolyl. Specific examples of bicyclic heteroaryl groups containing a six-membered ring fused to a five-membered ring include (but are not limited to) benzofuranyl, benzothiophenyl, benzimidazolyl, benzoxazolyl, isobenzoxazolyl, benzoisoazolyl, benzothiazolyl, benzoisothiazolyl, isobenzofuranyl, indolyl, isoindolyl, indolazinyl, purine (e.g., adenine, guanine), indazoleyl, pyrazolopyrimidinyl, triazolopyrimidinyl, and pyrazolopyridinyl. Specific examples of bicyclic heteroaryl groups containing two fused six-membered rings include (but are not limited to) quinolinyl, isoquinolinyl, pyridopyridinyl, quinoxalinyl, quinazolinyl, cenolinyl, phthalazinyl, naphthidyl, and pteridinyl. The specific heteroaryl group is those heteroaryl groups derived from thienyl, pyrroleyl, benzothienyl, benzofuranyl, indolyl, pyridyl, quinolinyl, imidazolyl, oxazolyl, and pyrazinyl.

[0068] The compounds of this invention can exist in specific geometric or stereoisomeric forms. This invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this invention.

[0069] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound of the present invention, 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 desired enantiomer in pure form. 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 salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0070] The term "stereoisomer" refers to compounds that have the same chemical structure but different spatial arrangements of atoms or groups. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans isomers), and hindered isomers, etc.

[0071] The terms "tautomer" or "tautomer form" refer 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 for the tautomer. For example, proton tautomers (also called prototropic 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. A specific example of a keto-enol tautomer is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one. Another example of tautomerism is phenol-keto tautomerism. A specific example of a phenol-keto tautomer is the interconversion between pyridine-4-ol and pyridine-4(1H)-keto. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0072] The term "racemate" refers to a mixture of two equimolar enantiomers that lack optical activity.

[0073] The term "pharmaceutically acceptable salt" refers to both organic and inorganic salts of the compounds of this invention. Pharmaceutically acceptable salts are well known in the field, as described in the literature: SMBerge et al., Pharmaceutical salts, J Pharm Sci. 1977 Jan; 66(1): 1-19. Salts formed from pharmaceutically acceptable non-toxic acids include, but are not limited to, inorganic acid salts formed by reaction with amino groups, such as hydrochlorides, hydrobroms, phosphates, sulfates, perchlorates, and organic acid salts such as acetates, decanoates, trifluoroacetates, oxalates, maleates, tartrates, citrates, succinates, malonates, or obtained by other methods described in the literature, such as ion exchange. Other pharmaceutically acceptable salts include adipate, malate, 2-hydroxypropionate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, transbutenedioic acid, glucono-heptahydrate, glycerophosphate, gluconate, hemisulfate, heptahydrate, hexanoate, hydroiodate, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pyruvate, pectinate, persulfate, 3-phenylpropionate, picrate, pentanoate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts obtained by means of appropriate bases include alkali metals, alkaline earth metals, ammonium, and nitrogen. + Salts of (C1-4 alkyl)4. This invention also envisions quaternary ammonium salts formed from any compound containing an N group. Water-soluble or oil-soluble or dispersed products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc. Pharmaceutically acceptable salts further include suitable, non-toxic ammonium, quaternary ammonium salts, and amine cations that resist the formation of equilibrium ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C1-8 sulfonates, and aromatic sulfonates.

[0074] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium capable of delivering an effective amount of the active substance of this invention without interfering with the biological activity of the active substance and without toxic side effects on the host or patient. Representative carriers include water, oil, vegetables and minerals, ointment bases, lotion bases, and ointment bases. These bases include suspending agents, thickeners, and transdermal penetration enhancers. Their formulations are well known to those skilled in the art of cosmetics or topical pharmaceuticals. For further information on carriers, see Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0075] The term "excipient" generally refers to the carrier, diluent, and / or medium required to formulate an effective pharmaceutical composition.

[0076] The terms “optional” or “optionally” refer to events or conditions that may occur but are not required to occur as described below, and the description includes both cases where said events or conditions occur and cases where said events or conditions do not occur.

[0077] The prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to be converted into the compounds of the present invention. Furthermore, the prodrugs can be converted into the compounds of the present invention in the in vivo environment via chemical or biochemical methods.

[0078] Some compounds of this invention may exist in non-solventized or solvated forms, including hydrated forms. Generally, solvated and non-solventized forms are equivalent and both are included within the scope of this invention.

[0079] The atoms in the compounds of this invention are isotopes. Isotope derivatization can typically prolong half-life, reduce clearance rate, stabilize metabolism, and enhance in vivo activity. Furthermore, one embodiment is included, wherein at least one atom is replaced by an atom having the same number of atoms (protons) but different mass numbers (protons and neutrons). Examples of isotopes included in the compounds of this invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, and chlorine atoms, each comprising... 2 H, 3 H, 13 C 14 C 15 N、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 36Cl. In particular, radioactive isotopes that emit radiation as they decay, such as 3 H or 14 C can be used for local anatomical examination of pharmaceutical preparations or compounds in vivo. Stable isotopes neither decay nor change with quantity and are not radioactive, therefore they can be used safely. When the atoms constituting the compounds of this invention are isotopes, the isotopes can be converted according to common methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0080] For example, the compounds of the present invention may contain atomic isotopes in non-natural proportions on one or more atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium. 2 H), Iodine-125 125 I) or C-14 14 C). All isotopic variations of the compounds of the present invention, regardless of radioactivity, are included within the scope of the present invention.

[0081] Furthermore, one or more hydrogen atoms in the compound of the present invention are coated with the isotope deuterium ( 2 The compounds of this invention, after being substituted with H), have the effects of prolonged half-life, reduced clearance rate, metabolic stabilization, and increased in vivo activity.

[0082] The preparation methods of the isotope derivatives typically include phase-transfer catalysis. For example, a preferred deuteration method employs a phase-transfer catalyst (e.g., tetraalkylammonium salt, NBu4HSO4). Using a phase-transfer catalyst to exchange the methylene protons of a diphenylmethane compound results in the introduction of higher levels of deuterium than reduction with deuterated silanes (e.g., triethyldeuterated silane) in the presence of an acid (e.g., methanesulfonic acid) or with Lewis acids such as aluminum trichloride using sodium deuterated borate.

[0083] For pharmaceuticals or pharmacologically active agents, the term "effective amount" or "therapeutic effective amount" refers to a sufficient quantity of a drug or agent that is non-toxic but achieves the desired effect. For the oral dosage forms of this invention, the "effective amount" of one active substance in the composition refers to the quantity required to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depending on the recipient's age and general condition, as well as the specific active substance. A suitable effective amount in any given case can be determined by a person skilled in the art through routine testing.

[0084] The terms “active ingredient,” “therapeutic agent,” “active substance,” or “active agent” refer to a chemical entity that can effectively treat a target disorder, disease, or symptom. Detailed Implementation

[0085] The present invention will be further described in detail below with reference to the embodiments, but the content of the invention is not limited to the embodiments.

[0086] The amino acids and intermediates thereof, resins and protecting groups, cleavage reagents, etc. of the present invention are commercially available or obtained according to the prior art, wherein Int1 is

[0087] Preferably,

[0088] The general route for intermediate AA4 is as follows:

[0089] In the route, X1 = amino protecting groups such as Boc, Cbz, Fmoc, Dmb, PMB, Bn, SEM, etc.

[0090] In the route, Y1 = carboxyl protecting groups such as t-Bu, methyl ester, ethyl ester, propargyl ester, TBS, and p-methoxybenzyl ester.

[0091] In the route, Z1 = leaving groups such as OTs, OTf, I, Br, Cl, F, etc.

[0092] Specifically, intermediate AA4:

[0093] Synthesis route:

[0094] Step 1: Synthesis of (2S,3S)-1-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid

[0095] (2S,3S)-3-hydroxypyrrolidine-2-carboxylic acid (1.5 g, 11.4 mmol) was dissolved in anhydrous dichloromethane (50 ml) at room temperature. Then Boc2O (3.5 g, 15.96 mmol) and DIPEA (4.4 g, 34.2 mmol) were added sequentially. The mixture was stirred at room temperature for 4 hours. After the reaction was completed, the product was concentrated to dryness to obtain an oily substance for the next step.

[0096] Step 2: Synthesis of di-tert-butyl(2S,3S)-3-hydroxypyrrolidine-1,2-dicarboxylic acid ester

[0097] (2S,3S)-1-(tert-butoxycarbonyl)-3-hydroxypyrrolidine-2-carboxylic acid (14.9 g, 64.4 mmol) was dissolved in 1 L of toluene at room temperature, followed by the addition of N,N-dimethylformamide di-tert-butyl acetal (52.7 g, 257.7 mmol). The mixture was stirred overnight at 85°C. After the reaction was complete, the reaction solution was concentrated directly, dissolved in water and ethyl acetate, extracted, and the organic phase was evaporated to dryness to obtain the crude product.

[0098] Step 3: Synthesis of di-tert-butyl(2S,3S)-3-(2-(allyloxy)-2-oxoethoxy)pyrrolidine-1,2-dicarboxylic acid ester

[0099] Di-tert-butyl(2S,3S)-3-hydroxypyrrolidine-1,2-dicarboxylic acid ester (12 g, 41.7 mmol) was dissolved in DMF (100 mL) at room temperature, followed by the addition of ethyl allyl 2-bromoacetate (22.4 g, 125.2 mmol) and cesium carbonate (40.8 g, 125.2 mmol). The mixture was stirred at 110°C for 5 hours. After the reaction was complete, the mixture was cooled to room temperature, and 5 L of water and 6 L of ethyl acetate were added to the reaction solution. The mixture was extracted, and the organic phase was stirred by rotary evaporation and passed through a normal phase (n-hexane:ethyl acetate = 100:0 to 4:1) to obtain a yellow oil.

[0100] Step 4: Synthesis of (2S,3S)-3-(2-(allyloxy)-2-oxoethoxy)pyrrolidine-2-carboxylic acid

[0101] Di-tert-butyl(2S,3S)-3-(2-(allyloxy)-2-oxoethoxy)pyrrolidine-1,2-dicarboxylic acid ester (7.5 g, 19.7 mmol) was dissolved in DCM (100 mL) at room temperature, followed by stirring overnight at room temperature with trifluoroacetic acid (22.2 g, 194.4 mmol). The next day, the reaction solution was directly evaporated to dryness. The crude product was then directly proceeded to the next step.

[0102] Step 5: Synthesis of (2S,3S)-1-(((9H-fluorene-9-yl)methoxy)carbonyl)-3-(2-(allyl)-2-oxoethoxy)pyrrolidine-2-carboxylic acid

[0103] (2S,3S)-3-(2-(allyloxy)-2-oxoethoxy)pyrrolidine-2-carboxylic acid (4.4 g, 19.1 mmol) was dissolved in THF (100 mL) and H2O (50 mL), and NaHCO3 (9.7 g, 115.2 mmol) was slowly added, followed by Fmoc-Cl (5.5 g, 21.2 mmol). The mixture was stirred at room temperature for 6 hours. The reaction solution was concentrated to remove THF, and then HCl was added to adjust the pH to 4. The mixture was extracted with ethyl acetate, and the organic phase was concentrated to dryness. The mixture was then passed through a normal phase (first hexane:DCM = 10:1 to 0:100, then DCM:MeOH = 0:100 to 10:1) to obtain the target product.

[0104] [M+H] + =452.3. 1H NMR (400MHz, DMSO-d6) δ13.08(s,1H),7.92(t,J=6.7Hz2H),7.72-7.61(m,2H ),7.48-7.29(m,4H),4.95(ddq,J=16.7,11.3,5.7Hz,1H),4.40-.5.30(m,1H ),5.25(dd,J=10.7,5.1Hz,1H),4.68-4.61(m,2H),4.40-2.11(m,6H),3.61- 3.42(m3H), 2.09(dt,J=13.9,6.8Hz,1H), 1.97(dtd,J=1.7,8.7,4.2Hz,1H).

[0105] Intermediate AA5:

[0106] Synthesis of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(1-allyl-5-fluoro-1H-indole-3-yl)propionic acid

[0107] Synthesis route:

[0108] Step 1: Synthesis of (S)-2-amino-3-(5-fluoro-1H-indol-3-yl)propionic acid

[0109] At room temperature, (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(5-fluoro-1H-indol-3-yl)propionic acid (7.80 g, 17.6 mmol, 1.00 eq) and diethylamine (2.57 g, 35.1 mmol, 3.62 mL, 2.00 eq) were added to DMF (3.00 mL), and the mixture was stirred at 20 °C for 2 hours.

[0110] LCMS showed that the reaction proceeds were completely reacted. The crude product was obtained by vacuum filtration, and then slurried in dichloromethane (30.0 ml * 3) at 20 °C. The filter cake was washed with 10.0 ml of dichloromethane and dried. The target product (S)-2-amino-3-(5-fluoro-1H-indol-3-yl)propionic acid (3.8 g) was obtained as a white solid.

[0111] Step 2: Synthesis of (S)-2-((tert-Butoxycarbonyl)amino)-3-(5-Fluoro-1H-indol-3-yl)propionic acid

[0112] At room temperature, (S)-2-amino-3-(5-fluoro-1H-indol-3-yl)propionic acid (3.80 g, 17.1 mmol, 1.00 eq), TEA (2.08 g, 20.5 mmol, 2.86 mL, 1.20 eq), and Boc2O (3.81 g, 17.4 mmol, 4.01 mL, 1.02 eq) were added to THF (200 mL) and H2O (200 mL), and the mixture was stirred at -5 to -0 °C for 5 hours.

[0113] LCMS showed that the starting material reaction was complete. 100 mL of 0.5 M hydrochloric acid aqueous solution was added to the reaction mixture, followed by extraction with ethyl acetate (250 mL * 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The target product (S)-2-((tert-butoxycarbonyl)amino)-3-(5-fluoro-1H-indol-3-yl)propionic acid (5.4 g) was obtained as a white solid.

[0114] Step 3: Synthesis of (S)-3-(1-allyl-5-fluoro-1H-indole-3-yl)-2-((tert-butoxycarbonyl)amino)propionic acid

[0115] At room temperature, NaH (1.75 g, 43.7 mmol, 60% purity, 3.00 eq) was added to THF (100 mL), and the mixture was cooled to 0 °C under nitrogen protection. Then (S)-2-((tert-butoxycarbonyl)amino)-3-(5-fluoro-1H-indol-3-yl)propionic acid (4.70 g, 14.6 mmol, 1.00 eq) was added, and the mixture was stirred at 0 °C for 330 minutes. Finally, allyl bromide (5.29 g, 4.7 mmol, 3.00 eq) was added, and the mixture was stirred at 25 °C for 5 hours.

[0116] LCMS showed that the reaction proceeds were complete. The reaction was quenched by slowly adding 100 ml of saturated ammonium chloride solution to the reaction mixture. The pH was adjusted to 5 with 1 M dilute hydrochloric acid solution, and the mixture was extracted with ethyl acetate (300 ml * 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The mixture was then purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) using silica gel. The target product, a yellow solid (S)-3-(1-allyl-5-fluoro-1H-indol-3-yl)-2-((tert-butyloxycarbonyl)amino)propionic acid (4.2 g), was obtained.

[0117] Step 4: Synthesis of (S)-3-(1-allyl-5-fluoro-1H-indole-3-yl)-2-aminopropionic acid

[0118] At room temperature, (S)-3-(1-allyl-5-fluoro-1H-indol-3-yl)-2-((tert-butoxycarbonyl)amino)propionic acid (5.1 g, 14.1 mmol, 1.00 eq) and 4M HCl / EtOAc (4 M, 52.8 mL, 15.0 eq) were added to DCM (50.0 mL), and the mixture was stirred at 25 °C for 2 hours.

[0119] LCMS showed that the reaction of the starting material was complete. The mixture was concentrated under reduced pressure to give (S)-3-(1-allyl-5-fluoro-1H-indol-3-yl)-2-aminopropionic acid (4.15 g), a pale yellow solid.

[0120] Step 5: Synthesis of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(1-allyl-5-fluoro-1H-indole-3-yl)propionic acid

[0121] At room temperature, (S)-3-(1-allyl-5-fluoro-1H-indol-3-yl)-2-aminopropionic acid (2.58 g, 8.64 mmol, 1.00 eq, HCl), Na3CO3 (2.29 g, 21.6 mmol, 1.06 mL, 2.50 eq) and FmocOSu (3.20 g, 9.50 mmol, 1.1 eq) were dissolved in DMF (30.0 mL) and H2O (45.0 mL), cooled to 0 °C, and stirred at 0 °C for 1 hour. The mixture was then heated to 25 °C and stirred for 3 hours.

[0122] LCMS showed that the reactants reacted completely. The pH of the reaction solution was adjusted to 5 with 1M hydrochloric acid aqueous solution, extracted with ethyl acetate (5ml*3), the organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The combined phases were purified by reversed-phase chromatography (column: Phenomenex luna C18 (250*70mm, 10um); mobile phase: [water(HCl)-ACN]; gradient: 30%-50% B over 30min), yielding the target product (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(1-allyl-5-fluoro-1H-indole-3-yl)propionic acid (5.2g).

[0123] LCMS RT=0.610min,m / z=485.1[M+H] +

[0124] 1H NMR:(400MHz,DMSO-d6)δ12.94(br d,J=1.8Hz,1H),7.88(d,J=7.5Hz,2H),7.71-7.52(m,3H),7.44-7.33(m,4H),7.27(td,J=7.4,14. 5Hz,2H),7.22(s,1H),6.95(dt,J=2.2,9.1Hz,1H),5.97-5.85(m,1H),5.16-4.88(m,2H),4.72(br d,J=5.1Hz,2H),4.26-4.09(m,4H),3.16(br dd,J=4.5,14.5Hz,1H),3.00(br dd, J = 9.2, 14.6 Hz, 1H).

[0125] Intermediate AA6:

[0126] Synthesis of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(((((allyloxy)carbonyl)amino)methyl)phenyl)propionic acid

[0127] Synthesis route:

[0128] Step 1: (S)-2-amino-3-(3-cyanophenyl)propionic acid

[0129] At 25°C, (S)-2-((tert-butoxycarbonyl)amino)-3-(3-cyanophenyl)propionic acid (25.0 g, 86.1 mmol, 1.00 eq) was dissolved in dichloromethane (50 mL), and hydrochloric acid / dioxane (25 mL) was added dropwise. The reaction was allowed to proceed for 2 hours. TLC (petroleum ether:ethyl acetate = 5:1) showed that the starting material disappeared and new spots formed. The reaction solution was concentrated to dryness to obtain the product (S)-2-amino-3-(3-cyanophenyl)propionic acid (16.0 g).

[0130] Step 2: (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-cyanophenyl)propionic acid

[0131] Dissolve (S)-2-amino-3-(3-cyanophenyl)propionic acid (16.0 g, 84.1 mol, 1.00 eq) and sodium bicarbonate (28.3 g, 336 mol, 4.00 eq) in dioxane (150 mL) and water (150 mL), cool to 0 °C, add 9-fluorene methyl ester (26.1 g, 101 mol, 1.20 eq), stir for 2 hours, heat to 25 °C, and react for 10 hours.

[0132] TLC (petroleum ether:ethyl acetate = 0:1) showed that the starting material disappeared and new spots were formed (R). f =0.51).

[0133] Dilute the reaction solution with 500 mL of water, extract with methyl tert-butyl ether (200 mL * 3), adjust the pH of the aqueous phase to about 3 with 1 N hydrochloric acid, extract with ethyl acetate (300 mL * 3), wash the organic phase with saturated brine, dry, filter, concentrate, and obtain the product (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-cyanophenyl)propionic acid (30.0 g).

[0134] Step 3: (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(aminomethyl)phenyl)propionic acid

[0135] At 25°C, (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-cyanophenyl)propionic acid (10.0 g, 24.3 mol, 1.00 eq) was dissolved in isopropanol (200 mL) and hydrochloric acid (50 mL). The reaction flask was gently purged with nitrogen, and Raney nickel (10.0 g, 117 mol, 4.81 eq) was added. The reaction was carried out under a nitrogen (15 Psi) atmosphere for 3 hours.

[0136] LCMS showed that the raw material disappeared, and the product was detected by MS (RT = 0.448 min).

[0137] Dilute with 300 mL of water and 600 mL of acetonitrile to clarify the solution, filter, concentrate, and freeze-dry to obtain the product (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(aminomethyl)phenyl)propionic acid (50 g, crude product).

[0138] Step 4: (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(((((allyloxy)carbonyl)amino)methyl)phenyl)propionic acid

[0139] (S)-2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(aminomethyl)phenyl)propionic acid (20.0 g, 44.2 mol, 1.00 eq) was dissolved in dioxane (200 mL) and water (200 mL), cooled to 0 °C, and then allyl chloroformate (5.85 g, 48.6 mol, 1.10 eq) and sodium carbonate (7.96 g, 75.1 mol, 1.70 eq) were added. The reaction was allowed to proceed for 0.5 hours.

[0140] The product was detected by LCMS (RT = 0.556 min).

[0141] Add 300 mL of water to the reaction solution, extract with ethyl acetate (100 mL * 3), filter, combine the organic phases, adjust the pH to approximately 7 with sodium bicarbonate aqueous solution, wash with saturated brine, dry and concentrate. The crude product is purified by reverse-phase (hydrochloric acid / methanol system) to obtain 2.89 g of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(((((allyloxy)carbonyl)amino)methyl)phenyl)propionic acid). Lyophilize to obtain 5.39 g of (S)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(3-(((((allyloxy)carbonyl)amino)methyl)phenyl)propionic acid. LCMS: RT = 0.548 min, MS (ESI) m / z = 501.1 [M+H] +

[0142] 1 H NMR: (400MHz, DMSO) δ7.88(d,J=7.5Hz,2H),7.77-7.70(m,2H),7.65(t,J=7.8Hz,2H),7.41(dt,J=2 .1,7.3Hz,2H),7.34-7.19(m,3H),7.18-7.07(m,3H),5.99-5.82(m,1H),5.33-5.09(m,2H),4.48(br d,J=5.3Hz,2H),4.26-4.10(m,6H),3.05(br dd,J=4.1,13.8Hz,1H),2.86(dd,J=10.4,13.7Hz,1H).

[0143] Linker-1:

[0144] Synthesis of 4-[but-3-enyl-[[4-[2-(9H-fluoren-9-ylmethoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid

[0145] Synthesis route:

[0146] Step 1: 4-[3-(tert-Butoxycarbonylamino)ethyl]benzoic acid

[0147] At 25°C, 4-(2-aminoethyl)benzoic acid (14 g.0, 1.00 eq, hydrochloride) was dissolved in tetrahydrofuran (140 mL) and water (140 mL), and Boc2O (18.2 g, 1.20 eq) and triethylamine (15.5 g, 2.20 eq) were added, and the reaction was carried out for 2 hours.

[0148] LCMS showed the MS value of the product. The reaction solution was adjusted to pH 3 to 5 with saturated citric acid aqueous solution, 1000 mL of water was added, and extraction was performed with ethyl acetate (500 mL * 3). The concentrated organic phase was washed with saturated brine (400 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to give 4-[2-(tert-butoxycarbonylamino)ethyl]benzoic acid (18.0 g).

[0149] Step 2: tert-butyl N-[2-[4-[methoxy(methyl)carbamoyl]phenyl]ethyl]carbamate

[0150] 4-[2-(tert-Butoxycarbonylamino)ethyl]benzoic acid (18.0 g, 1.00 eq) was dissolved in THF (120 mL) at 20 °C, followed by the addition of 2-chloro-4,6-dimethoxy-1,3,5-triazine (14.3 g, 81.4 mmol, 1.20 eq) and N-methylmorpholine (20.6 g, 3.00 eq). After the addition was complete, the mixture was stirred at 20 °C for one hour. Then, N-methoxymethylamine hydrochloride (6.62 g, 67.8 mmol, 1.00 eq) was added to the solution, and the reaction was carried out at 20 °C for 12 hours.

[0151] LCMS showed the MS value of the product. The reaction solution was diluted with water (800 mL) and extracted with ethyl acetate (200 mL * 3). The concentrated organic phase was washed with saturated brine (200 mL * 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse-phase preparation (HCl condition) to obtain tert-butyl N-[2-[4-[methoxy(methyl)carbamoyl]phenyl]ethyl]carbamate (14.0 g).

[0152] Step 3: N-[2-(4-formylphenyl)ethyl]carbamate tert-butyl ester

[0153] 13.5 g, 43.7 mmol, 1.00 eq of tert-butyl N-[2-[4-[methoxy(methyl)carbamoyl]phenyl]ethyl]carbamate (13.5 g, 43.7 mmol, 1.00 eq) was dissolved in 100 mL of THF. DIBAL-H (1.00 M, 87.5 mL, 2.00 eq) was added dropwise at -70 °C. After the addition was complete, the reaction was maintained at this temperature for 1 hour.

[0154] TLC (petroleum ether:ethyl acetate = 1:1) showed that the reaction of the starting materials was complete. The reaction solution was quenched with a saturated sodium bicarbonate aqueous solution (400 mL), followed by extraction with ethyl acetate (60 mL * 3). The concentrated organic phase was washed with saturated brine (50 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the product N-[2-(4-formylphenyl)ethyl]carbamate tert-butyl ester (10.0 g).

[0155] Step 4: tert-butylN-[2-[4-[(but-3-enylamino)methyl]phenyl]ethyl]carbamic acid

[0156] At 20°C, but-3-ene-1-amine hydrochloride (6.47 g, 1.50 eq) and sodium acetate (4.94 g, 60.2 mmol, 1.50 eq) were dissolved in dichloromethane (20 mL) and stirred for 1 hour. Then, N-[2-(4-formylphenyl)ethyl]carbamate tert-butyl ester (10.0 g, 40.1 mmol, 1.00 eq) and acetic acid (2.44 g, 1.01 eq) were added to the solution and stirred for 1 hour. Sodium borohydride acetate (21.3 g, 2.50 eq) was added in portions to the above solution, and the reaction was maintained at this temperature for 1 hour after the addition was complete.

[0157] TLC (petroleum ether:ethyl acetate = 1:1) showed that the starting material was completely consumed. The reaction solution was quenched with saturated sodium bicarbonate aqueous solution (300 mL), followed by extraction with dichloromethane (50 mL * 4). The concentrated organic phase was washed with saturated brine (40 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse-phase preparation (HCl codition) to obtain the product tert-butylN-[2-[4-[(but-3-enylamino)methyl]phenyl]ethyl]carbamic acid (6.20 g).

[0158] Step 5: 4-[but-3-enyl-[[4-[2-(tert-butoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid

[0159] tert-butyl N-[2-[4-[(but-3-enylamino)methyl]phenyl]ethyl]carbamic acid (6.20 g, 20.4 mmol, 1.00 eq) and tetrahydrofuran-2,5-dione (3.06 g, 30.55 mmol, 1.5 eq) were dissolved in THF (40 mL) and stirred at 20 °C for 0.5 hours.

[0160] TLC (petroleum ether:ethyl acetate = 1:0) showed that the starting material was consumed. Water (200 mL) was added to the reaction solution, followed by extraction with dichloromethane (50 mL * 3). The concentrated organic phase was washed with saturated brine (50 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 10:1 to 0:1) to obtain the product 4-[but-3-enyl-[[4-[2-(tert-butoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid (8.00 g).

[0161] Step 6: 4-[[4-(2-aminoethyl)phenyl]methyl-but-3-enyl-amino]-4-oxo-butyric acid

[0162] At 20°C, 4-[but-3-enyl-[[4-[2-(tert-butoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid (8.00 g, 19.8 mmol, 1.00 eq) and ethyl hydrochloride (4.00 M, 8.09 eq) were dissolved in dichloromethane (40 mL) and stirred for 1 hour.

[0163] TLC (petroleum ether: ethyl acetate = 1:1) showed that the reaction of the raw materials was complete. The reaction solution was concentrated to obtain 6.80 g of 4-[[4-(2-aminoethyl)phenyl]methyl-but-3-enyl-amino]-4-oxo-butyric acid.

[0164] Step 7: 4-[but-3-enyl-[[4-[2-(9H-fluoren-9-ylmethoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid

[0165] At 20 °C, 4-[[4-(2-aminoethyl)phenyl]methyl-but-3-enyl-amino]-4-oxo-butyric acid (6.80 g, 22.3 mmol, 1.00 eq) and sodium carbonate (5.92 g, 55.8 mmol, 2.50 eq) were dissolved in dioxane (80 mL) and water (80 mL), and FmocOSu (8.29 g, 24.6 mmol, 1.10 eq) was added in portions, followed by reaction at room temperature for 1 hour.

[0166] LCMS showed the MS value of the product. The reaction solution was adjusted to pH 3-5 with 1M citric acid, followed by extraction with dichloromethane (50 mL * 4). The concentrated organic phase was washed with saturated brine (50 mL * 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by reverse-phase chromatography (column: Phenomenex luna C18 (250 * 70 mm, 10 μm); mobile phase: [water (HCl)-ACN]; gradient: 30%-75% B over 22 min) to obtain 5.8 g of product 4-[but-3-enyl-[[4-[2-(9H-fluorene-9-ylmethoxycarbonylamino)ethyl]phenyl]methyl]amino]-4-oxobutyric acid.

[0167] LCMS RT=0.569min,MS(ESI)m / z=527.4[M+H] +

[0168] 1H NMR: (400MHz, MeOD): δ7.73-7.66(m,2H),7.54-7.46(m,2H),7.33(t,J=7.6Hz,2H),7.28-7.21(m,2H),7.13-6.93(m,4H),5.75-5.58(m,1H), 5.06-4.92(m,2H),4.79-4.67(m,1H),4.58-4.40(m,2H),4.55-4.31(m ,1H),4.19-4.08(m,1H),3.46-3.17(m,4H),2.79-2.51(m,6H),2.24(br s,1H).

[0169] Example 1: Synthesis of compounds A1-A7

[0170] Synthesis route:

[0171] Synthesis of compound A2

[0172] Synthesis route:

[0173] Solid-phase synthesis of compound A2-1

[0174] Peptides were synthesized using standard Fmoc chemical synthesis.

[0175] 1) Resin preparation: AA1 (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g) and DIEA (4.00 mmol, 4.00 eq) from DCM (30.0 mL) were added to 2-CTC resin (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g). The mixture was stirred with N2 at 20 °C for 2 hours, then MeOH (2.00 mL) was added and stirred again with N2 for 30.0 minutes. The resin was washed with DMF (30.0 mL * 5).

[0176] 2) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0177] 3) Coupling with AA2: Add a DMF (30.0 mL) solution of AA2 (3.00 mmol, 3.00 eq) and HATU (2.85 mmol, 2.85 eq) to the resin, then add DIEA (6.00 mmol, 6.00 eq), and stir the resin with N2 for 2 h.

[0178] 4) Repeat steps 2 and 3 above for the coupling of the following amino acids: (AA3-AA6)

[0179] 5) Deprotection: Add DCM (50.0 mL), PhSiH3 (10.0 mmol, 10.0 eq), and Pd(PPh3)4 (0.10 mmol, 0.10 eq), and stir with N2 at room temperature for 30.0 min*3. Then wash the resin with DMF (50.0 mL*5).

[0180] 6) Cycloning: Add HOAt (3.00 mmol, 3.00 eq) in DMF (50.0 mL), add DIC (3.00 mmol / L, 3.00 equivalent), and stir the mixture with N2 at 25 °C for 12 hours. Wash the resin with DMF (30 mL * 5).

[0181] 7) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin again with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0182] 8) Repeat steps 2 and 3 above for the coupling of the following amino acids: (AA7-AA8)

[0183] 9) Linker Coupling: A solution of linker (1.150 mmol, 1.50 eq) and HOAt (1.50 mmol, 1.50 eq) in DMF (30.0 mL) was added to the resin, followed by the addition of DIC (6.00 mmol, 6.00 eq). The mixture was stirred with N2 at 25 °C for 12 hours. The resin was washed with DMF (30 mL * 5).

[0184] 10) Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0185] 11) Cleavage: The resin was washed with MeOH (50.0 mL * 3) and dried under vacuum to obtain 3.70 g of peptide resin. The peptide resin was then treated with cleavage mixture (60.0 mL) (20% HFIP in DCM) for 30.0 min * 3, filtered and concentrated under reduced pressure to obtain crude product A2-1 (1.3 g).

[0186] definition:

[0187] The amino acids and linkers used in this peptide are as follows:

[0188] AA1:Fmoc-L-2-methylazacyclobutane-2-carboxylic acid

[0189] AA2:Fmoc-p-methoxy-L-phenylalanine

[0190] AA3:Fmoc-O-tert-butyl-L-threonine

[0191] AA4:AA4-0

[0192] AA5:AA5-0

[0193] AA6:AA6-0

[0194] AA7: N2-fluorenylmethoxycarbonyl-N3-tert-butyloxycarbonyl-D-2,3-diaminopropionic acid

[0195] AA8:Fmoc-L-alanine

[0196] Linker: Linker-0

[0197] Side chain: side chain

[0198] Synthesis of compound A2-2

[0199] Compound A2-1 (1.3 g, 0.8 mmol) was dissolved in DCM (1 L), and HOBt (0.22 g, 1.6 mmol, 2.00 eq), DIEA (0.21 g, 1.6 mmol, 2.00 eq), and TBTU (0.51 g, 1.6 mmol, 2.00 eq) were added. The mixture was then reacted at room temperature for 3 h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL * 4). The organic phases were combined, concentrated, and purified to obtain compound A2-2 (180 mg).

[0200] Synthesis of compound A2-3

[0201] Under nitrogen protection, AcOH (8.04 g, 133 mmol, 7.66 mL, 1162 eq) and Zhan catalyst 1B (50.7 mg, 69.1 μmol, 0.60 eq) were added to a DCM (120 mL) solution of compound A2-2 (180 mg, 115 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to obtain a residue. The residue was purified by column chromatography (DCM:methanol = 10:1) to give a brown solid compound A2-3 (100 mg).

[0202] Synthesis of compound A2-4

[0203] Pd / C (8.00 mg, 10% purity) was added to a 4 mL THF solution of A2-3 (100 mg, 65.1 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated, and column chromatography (n-hexane:ethyl acetate = 10:1) gave a white solid A2-4 (90.0 mg).

[0204] Synthesis of compound A2-5

[0205] TFA (2.06 g, 3.61 mmol, 1.34 mL, 20% purity, 61.7 eq) was added to a DCM (5 mL) solution of A2-4 (90.0 mg, 58.5 μmol, 1.00 eq), and the mixture was stirred at 25 °C for 2 hours. The solution was concentrated, and then 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness, and then 0.2 mL of a 4M HCl solution of dioxane and 10 mL of DCM were added. The mixture was evaporated to dryness, and then 10 mL of DCM and 4 mL of toluene were added again. The mixture was evaporated to dryness to give a white solid product A2-5 (70 mg).

[0206] Synthesis of compound A2

[0207] To a DMF (2.00 mL) solution of A2-5 (70.0 mg, 49.3 μmol, 1.00 eq, HCl) and Int1 (25.2 mg, 98.7 μmol, 2.00 eq, HBr), DIEA (12.7 mg, 98.7 μmol, 16.3 μL, 2.00 eq) and AOP (35.0 mg, 79.0 μmol, 1.60 eq) were added. The mixture was stirred at 25 °C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was lyophilized and repurified by preparative HPLC (0.01% HCl) to give A2 (19 mg), LCMS: 1537.

[0208] Example 2: Synthesis of compounds B1-B11

[0209] Synthesis route:

[0210] Synthesis of compound B7

[0211] Synthetic route

[0212] solid phase synthesis

[0213] Peptides were synthesized using standard Fmoc chemical synthesis.

[0214] 1) Resin preparation: BB1 (1.00 mmol, 1.00 eq), and DIEA (4.00 mmol, 4.00 eq) from DCM (30.0 mL) were added to 2-CTC resin (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g). The mixture was stirred with N2 at 20 °C for 2 hours, then MeOH (2.00 mL) was added and stirred again with N2 for 30.0 minutes. The resin was washed with DMF (30.0 mL * 5).

[0215] 2) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0216] 3) Coupling with BB2: Add a DMF (30.0 mL) solution of BB2 (3.00 mmol, 3.00 eq) and HATU (2.85 mmol, 2.85 eq) to the resin, then add DIEA (6.00 mmol, 6.00 eq), and stir the resin with N2 for 2 h.

[0217] 4) Repeat steps 2 and 3 above for the coupling of the following amino acids: (BB3-BB6)

[0218] 5) Deprotection: Add DCM (50.0 mL), PhSiH3 (10.0 mmol, 10.0 eq), and Pd(PPh3)4 (0.10 mmol, 0.10 eq), and stir with N2 at room temperature for 30.0 min*3. Then wash the resin with DMF (50.0 mL*5).

[0219] 6) Cycloning: Add HOAt (3.00 mmol, 3.00 eq) in DMF (50.0 mL), add DIC (3.00 mmol / L, 3.00 equivalent), and stir the mixture with N2 at 25 °C for 12 hours. Wash the resin with DMF (30 mL * 5).

[0220] 7) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin again with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0221] 8) Repeat steps 2 and 3 above for the coupling of the following amino acids: (BB7-BB8)

[0222] 9) Linker Coupling: A solution of linker (1.150 mmol, 1.50 eq) and HOAt (1.50 mmol, 1.50 eq) in DMF (30.0 mL) was added to the resin, followed by the addition of DIC (6.00 mmol, 6.00 eq). The mixture was stirred with N2 at 25 °C for 12 hours. The resin was washed with DMF (30 mL * 5).

[0223] 10) Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0224] 11) Cleavage: The resin was washed with MeOH (50.0 mL * 3) and dried under vacuum to obtain 3.70 g of peptide resin. The peptide resin was then treated with cleavage mixture (60.0 mL) (20% HFIP in DCM) for 30.0 min * 3, filtered and concentrated under reduced pressure to obtain crude product B7-1 (1.3 g).

[0225] definition:

[0226] The amino acids and linkers used in this peptide are as follows:

[0227] BB1:Fmoc-α-methyl-L-proline

[0228] BB2:Fmoc-L-3,4-Dimethoxyphenylalanine

[0229] BB3:Fmoc-O-tert-butyl-L-threonine

[0230] BB4:BB4-0

[0231] BB5:BB5-0

[0232] BB6:BB6-0

[0233] BB7:N2-fluorenylmethoxycarbonyl-N3-tert-butyloxycarbonyl-D-2,3-diaminopropionic acid

[0234] BB8:Fmoc-L-alanine

[0235] Linker: Linker-0

[0236] Side chain: side chain

[0237] Synthesis of compound B7-2

[0238] Compound B7-1 (0.7 g, 0.43 mmol) was dissolved in DCM (1 L), and HOBt (0.276 g, 0.86 mmol, 2.00 eq), DIEA (0.22 g, 1.72 mmol, 4.00 eq), and TBTU (0.12 g, 0.86 mmol, 2.00 eq) were added. The mixture was then reacted at room temperature for 3 h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL * 4). The organic phases were combined, concentrated, and purified to give compound B7-2 (118 mg, 17.1% yield).

[0239] Synthesis of compound B7-3

[0240] Under nitrogen protection, AcOH (5.8 g, 98 mmol, 5.62 mL, 1345 eq) and Grubbs II catalyst (CAS246047-72-3) (37 mg, 44 μmol, 0.60 eq) were added to a DCE (150 mL) solution of compound 2 (118 mg, 73 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to give a residue. The residue was purified by column chromatography (DCM:methanol = 10:1) to give a brown solid compound B7-3 (115 mg).

[0241] Synthesis of compound B7-4

[0242] Pd / C (100 mg, 950 μmol, 10% purity) was added to a methanol (20 mL) solution of B7-3 (115 mg, 95 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated to give a white solid B7-4 (115.0 mg, crude).

[0243] Synthesis of compound B7-5

[0244] TFA (3.07 g, 26.9 mmol, 283 eq) was added to a DCM (5 mL) solution of B7-4 (115 mg, 95 μmol, 1.00 eq), and the mixture was stirred at 25 °C for 2 hours. The solution was concentrated, and then 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness, and then 0.2 mL of a 4M HCl solution of dioxane and 10 mL of DCM were added. The mixture was evaporated to dryness, and then another 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness again to give a white solid product B7-5 (115 mg).

[0245] Synthesis of compound B7

[0246] To a DMF (5.00 mL) solution of B7-5 (97 mg, 68 μmol, 1.00 eq) and Int1 (30 mg, 170 μmol, 2.50 eq, HBr), DIEA (52 mg, 410 μmol, 6.00 eq) and Carter's condensing agent (75 mg, 170 μmol, 2.50 eq) were added. The mixture was stirred at 25 °C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was lyophilized and repurified by preparative HPLC (0.01% HCl) to give B7 (32.4 mg), LCMS: 1581.

[0247] Example 3 Synthesis of compounds C1-C3

[0248] Synthesis route:

[0249] Example 4: Synthesis of the compound

[0250] Synthesis route:

[0251] Wherein: the synthesis of D1

[0252] Synthesis route:

[0253] Synthesis of compound D1-2

[0254] Compound D1-1 (1.3 g, 0.807 mmol) was dissolved in DCM (1.30 L), and HOBt (1.61 mmol, 2.00 eq), DIEA (4.039 mmol, 5.00 eq), and TBTU (1.61 mmol, 2.00 eq) were added. The reaction was then carried out at room temperature for h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL * 4). The organic phases were combined, concentrated, and purified to give compound D1-2 (255 mg, 160 μmol, 19.9% ​​yield). [M+H] + =1591

[0255] Synthesis of compound D1-3.

[0256] Under nitrogen protection, AcOH (10.3 g, 172 mmol, 9.89 mL, 1194 eq) and Zhan catalyst 1B (70.5 mg, 96.1 μmol, 0.60 eq) were added to a DCE (230 mL) solution of compound D1-2 (230 mg, 144 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to give a residue. The residue was purified by preparative high-performance liquid chromatography (TFA) to give a white solid compound D1-3 (170 mg), [M+H). + =1563.

[0257] Synthesis of compound D1-4

[0258] The solution of D1-3 (170 mg, 108 μmol, 1.00 eq) in TFA (3.36 g, 14.7 mmol, 2.00 mL, 50% purity, 135 eq) was added, and the mixture was stirred at 25 °C for 1 hour. The mixture was concentrated, and then the reaction mixture was directly purified by preparative HPLC (TFA) to give a white solid D2-4 (92.0 mg).

[0259] Synthesis of compound D1-5

[0260] Pd / C (9.20 mg, 8.645 μmol, 10% purity) was added to a THF (6.00 mL) solution of D1-4 (92.0 mg, 65.4 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated to give a white solid D1-5 (70.0 mg, crude), [M+H]. + =1409.

[0261] Synthesis of compound D1

[0262] Add DIEA (25.6 mg, 198 μmol, 32.8 μL, 4.00 eq) and AOP (35.2 mg, 79.5 μmol, 1.60 eq) to a DMF (2.60 mL) solution of D1-5 (70.0 mg, 49.6 μmol, 1.00 eq) and Int1 (25.3 mg, 99.3 μmol, 2.60 eq, HBr) to a solution of D1-5 (70.0 mg, 49.6 μmol, 1.00 eq) and HBr (2.60 mL ...

[0263] The mixture was stirred at 25°C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was freeze-dried and repurified by preparative HPLC (0.01% HCl) to give D1 (14.0 mg), LCMS: 1564.0.

[0264] Synthesis of D3

[0265] Synthesis route:

[0266] Synthesis of compound D3-2

[0267] Compound D3-1 (1.3 g, 0.8 mmol) was dissolved in DCM (1 L), and HOBt (0.22 g, 1.6 mmol, 2.00 eq), DIEA (0.21 g, 1.6 mmol, 2.00 eq), and TBTU (0.51 g, 1.6 mmol, 2.00 eq) were added. The mixture was then reacted at room temperature for 3 h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL x 4). The organic phases were combined, concentrated, and purified to obtain compound D3-2 (330 mg, 25.7% yield), [M+H]. + =1621.

[0268] Synthesis of compound D3-3.

[0269] Under nitrogen protection, AcOH (4.0 g, 67 mmol, 3.85 mL, 1345 eq) and Grubbs II catalyst (CAS246047-72-3) (25 mg, 30 μmol, 0.60 eq) were added to a DCE (80 mL) solution of compound D3-2 (80 mg, 50 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to give a residue. The residue was purified by column chromatography (DCM:methanol = 10:1) to give a brown solid compound D3-3 (70 mg), [M+H). + =1603.

[0270] Synthesis of compound D3-4

[0271] Pd / C (47 mg, 440 μmol, 10% purity) was added to a methanol (40 mL) solution of D3-3 (70 mg, 44 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated to give a white solid D3-4 (70.0 mg, crude), [M+H]. + =1575.

[0272] Synthesis of compound D3-5

[0273] TFA (2.26 g, 19.8 mmol, 450 eq) was added to a DCM (5 mL) solution of D3-4 (70 mg, 44 μmol, 1.00 eq), and the mixture was stirred at 25 °C for 2 hours. The solution was concentrated, and then 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness, and then 0.2 mL of a 4M HCl solution of dioxane and 10 mL of DCM were added. The mixture was evaporated to dryness, and then 10 mL of DCM and 4 mL of toluene were added again. The mixture was evaporated to dryness to give a white solid product D3-5 (64 mg).

[0274] Synthesis of compound D3

[0275] To a DMF (5.00 mL) solution of D3-5 (40 mg, 27 μmol, 1.00 eq) and Int1 (5 mg, 30 μmol, 1.10 eq, HBr), DIEA (17 mg, 140 μmol, 5.00 eq) and HATU (12 mg, 32 μmol, 1.20 eq) were added. The mixture was stirred at 25 °C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was lyophilized and repurified by preparative HPLC (0.01% HCl) to give D3 (14.4 mg), LCMS: [0.5 M + H₂]. + =788.6.

[0276] Example 5: Synthesis of compounds E1-E7

[0277] Synthesis route:

[0278] Synthesis of compound E1

[0279] Synthetic route

[0280] solid phase synthesis

[0281] Peptides were synthesized using standard Fmoc chemical synthesis.

[0282] 1) Resin preparation: AA1 (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g) and DIEA (4.00 mmol, 4.00 eq) from DCM (30.0 mL) were added to 2-CTC resin (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g). The mixture was stirred with N2 at 20 °C for 2 hours, then MeOH (2.00 mL) was added and stirred again with N2 for 30.0 minutes. The resin was washed with DMF (30.0 mL * 5).

[0283] 2) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0284] 3) Coupling with EE2: Add a DMF (30.0 mL) solution of EE2 (3.00 mmol, 3.00 eq) and HATU (2.82 mmol, 2.85 eq) to the resin, then add DIEA (6.00 mmol, 6.00 eq), and stir the resin with N2 for 2 h.

[0285] 4) Repeat steps 2 and 3 above for the coupling of the following amino acids: (EE3-EE6)

[0286] 5) Deprotection: Add DCM (50.0 mL), PhSiH3 (10.0 mmol, 10.0 eq), and Pd(PPh3)4 (0.10 mmol, 0.10 eq), and stir with N2 at room temperature for 30.0 min*3. Then wash the resin with DMF (50.0 mL*5).

[0287] 6) Cycloning: Add HOAt (3.00 mmol, 3.00 eq) in DMF (50.0 mL), add DIC (3.00 mmol / L, 3.00 equivalent), and stir the mixture with N2 at 25 °C for 12 hours. Wash the resin with DMF (30 mL * 5).

[0288] 7) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin again with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0289] 8) Repeat steps 2 to 3 above for the coupling of the following amino acids: (EE7-EE8)

[0290] 9) Linker Coupling: A solution of linker (1.150 mmol, 1.50 eq) and HOAt (1.50 mmol, 1.50 eq) in DMF (30.0 mL) was added to the resin, followed by the addition of DIC (6.00 mmol, 6.00 eq). The mixture was stirred with N2 at 25 °C for 12 hours. The resin was washed with DMF (30 mL * 5).

[0291] 10) Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0292] 11) Cleavage: The resin was washed with MeOH (50.0 mL * 3) and dried under vacuum to obtain 3.70 g of peptide resin. The peptide resin was then treated with cleavage mixture (60.0 mL) (20% HFIP in DCM) for 30.0 min * 3, filtered and concentrated under reduced pressure to obtain crude product E1-1 (1.3 g).

[0293] definition:

[0294] The amino acids and linkers used in this peptide are as follows:

[0295] EE1:Fmoc-L-α-methyl-proline

[0296] EE2:Fmoc-p-methoxy-L-phenylalanine

[0297] EE3:Fmoc-O-tert-butyl-L-threonine

[0298] EE4:EE4-0

[0299] EE5:EE5-0

[0300] EE6:EE6-0

[0301] EE7: N2-fluorenylmethoxycarbonyl-N3-tert-butyloxycarbonyl-D-2,3-diaminopropionic acid

[0302] EE8: Fmoc-L-alanine

[0303] Linker: Linker-1

[0304] Side chain: side chain

[0305] Synthesis of compound E1-2

[0306] Compound E1-1 (1.3 g, 0.8 mmol) was dissolved in DCM (1 L), and HOBt (0.22 g, 1.6 mmol, 2.00 eq), DIEA (0.21 g, 1.6 mmol, 2.00 eq), and TBTU (0.51 g, 1.6 mmol, 2.00 eq) were added. The mixture was then reacted at room temperature for 3 h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL * 4). The organic phases were combined, concentrated, and purified to obtain compound E1-2 (160 mg, 10% yield).

[0307] Synthesis of compound E1-3.

[0308] Under nitrogen protection, AcOH (5.78 g, 96.3 mmol, 5.51 mL, 1179 eq) and Zhan catalyst 1B (35.9 mg, 49.0 μmol, 0.60 eq) were added to a DCM (130 mL) solution of compound E1-2 (130 mg, 81.7 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to give a residue. The residue was purified by column chromatography (DCM:methanol = 10:1) to give a brown solid compound E1-3 (100 mg).

[0309] Synthesis of compound E1-4

[0310] Pd / C (12.0 mg, 10% purity) was added to a 6 mL THF solution of E1-3 (100 mg, 63.3 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated, and column chromatography (n-hexane:ethyl acetate = 10:1) gave a white solid E1-4 (95.0 mg, crude).

[0311] Synthesis of compound E1-5

[0312] TFA (1.69 g, 2.97 mmol, 1.10 mL, 20% purity, 48.9 eq) was added to a DCM (5 mL) solution of E1-4 (95.0 mg, 60.7 μmol, 1.00 e) and stirred at 25 °C for 2 hours. The solution was concentrated, and then 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness, and then 0.2 mL of a 4M HCl solution of dioxane and 10 mL of DCM were added. The mixture was evaporated to dryness, and then 10 mL of DCM and 4 mL of toluene were added again. The mixture was evaporated to dryness to give a white solid product E1-5 (87 mg).

[0313] Synthesis of compound E1-6

[0314] To a DMF (2.00 mL) solution of E1-5 (87.0 mg, 60.2 μmol, 1.00 eq, HCl) and Int1 (30.7 mg, 120 μmol, 2.00 eq, HBr), DIEA (31.1 mg, 240 μmol, 39.8 μL, 4.00 eq) and AOP (42.6 mg, 96.3 μmol, 1.60 eq) were added. The mixture was stirred at 25 °C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was lyophilized and repurified by preparative HPLC (0.01% HCl) to give E1 (17 mg), LCMS [M].+ :1565.

[0315] Example 6 Synthesis of compounds F1-F2

[0316] Synthesis route:

[0317] Among them, R 71 R 73 Same as R7 and R8 definitions, R 72 Selected from C 1-6 alkyl or alkyl-substituted C 1-6 Alkyl groups.

[0318] Synthesis of compound F1

[0319] solid phase synthesis

[0320] Peptides were synthesized using standard Fmoc chemical synthesis.

[0321] 1) Resin preparation: FF1 (1.00 mmol, 1.00 eq), and DIEA (4.00 mmol, 4.00 eq) from DCM (30.0 mL) were added to 2-CTC resin (1.00 mmol, 1.00 eq, Sub: 0.50 mmol / g). The mixture was stirred with N2 at 20 °C for 2 hours, then MeOH (2.00 mL) was added and stirred again with N2 for 30.0 minutes. The resin was washed with DMF (30.0 mL * 5).

[0322] 2) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0323] 3) Coupling with FF2: Add a DMF (30.0 mL) solution of FF2 (3.00 mmol, 3.00 eq) and HATU (2.85 mmol, 2.85 eq) to the resin, then add DIEA (6.00 mmol, 6.00 eq), and stir the resin with N2 for 2 h.

[0324] 4) Repeat steps 2 and 3 above for the coupling of the following amino acids: (FF3-FF6)

[0325] 5) Deprotection: Add DCM (50.0 mL), PhSiH3 (10.0 mmol, 10.0 eq), and Pd(PPh3)4 (0.10 mmol, 0.10 eq), and stir with N2 at room temperature for 30.0 min*3. Then wash the resin with DMF (50.0 mL*5).

[0326] 6) Cycloning: Add HOAt (3.00 mmol, 3.00 eq) in DMF (50.0 mL), add DIC (3.00 mmol / L, 3.00 equivalent), and stir the mixture with N2 at 25 °C for 12 hours. Wash the resin with DMF (30 mL * 5).

[0327] 7) Deprotection: Add 30.0 mL of 20% piperidine DMF solution and stir the resin again with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0328] 8) Repeat steps 2 and 3 above for the coupling of the following amino acids: (FF7-FF8)

[0329] 9) Linker Coupling: A solution of linker (1.150 mmol, 1.50 eq) and HOAt (1.50 mmol, 1.50 eq) in DMF (30.0 mL) was added to the resin, followed by the addition of DIC (6.00 mmol, 6.00 eq). The mixture was stirred with N2 at 25 °C for 12 hours. The resin was washed with DMF (30 mL * 5).

[0330] 10) Add 30.0 mL of 20% piperidine DMF solution and stir the resin with N2 for 15.0 minutes. Wash the resin with DMF (30.0 mL * 5) and filter to obtain the resin.

[0331] 11) Cleavage: The resin was washed with MeOH (50.0 mL * 3) and dried under vacuum to obtain 3.70 g of peptide resin. The peptide resin was then treated with cleavage mixture (60.0 mL) (20% HFIP in DCM) for 30.0 min * 3, filtered and concentrated under reduced pressure to obtain crude product F1-1 (1.0 g).

[0332] definition:

[0333] The amino acids and linkers used in this peptide are as follows:

[0334] FF1:Fmoc-α-methyl-L-proline

[0335] FF2:Fmoc-p-methoxy-L-phenylalanine

[0336] FF3:Fmoc-O-tert-butyl-L-threonine

[0337] FF4:FF4-0

[0338] FF5:FF5-0

[0339] FF6:FF6-0

[0340] FF7:N2-fluorenylmethoxycarbonyl-N3-tert-butyloxycarbonyl-N3-methyl-D-2,3-diaminopropionic acid

[0341] FF8: Fmoc-L-alanine

[0342] Linker: Linker-0

[0343] Side chain: side chain

[0344] Synthesis route:

[0345] Synthesis of compound F1-2

[0346] Compound F1-1 (1.0 g, 0.62 mmol) was dissolved in DCM (1 L), and HOBt (0.17 g, 1.24 mmol, 2.00 eq), DIEA (0.16 g, 1.24 mmol, 2.00 eq), and TBTU (0.40 g, 1.24 mmol, 2.00 eq) were added. The mixture was then reacted at room temperature for 3 h. 200 mL of saturated sodium bicarbonate was added, and the mixture was extracted with DCM (100 mL * 4). The organic phases were combined, concentrated, and purified to obtain compound F1-2 (300 mg, 30% yield).

[0347] Synthesis of compound F1-3.

[0348] Under nitrogen protection, AcOH (4.6 g, 76 mmol, 4.38 mL, 1345 eq) and Grubbs II catalyst (CAS246047-72-3) (29 mg, 34 μmol, 0.60 eq) were added to a DCE (80 mL) solution of compound F1-2 (90 mg, 57 μmol, 1.00 eq). The reaction mixture was stirred at 55 °C for 2 hours. The reaction mixture was concentrated to give a residue. The residue was purified by column chromatography (DCM:methanol = 10:1) to give a brown solid compound F1-3 (88 mg).

[0349] Synthesis of compound F1-4

[0350] Pd / C (60 mg, 560 μmol, 10% purity) was added to a methanol (40 mL) solution of F1-3 (88 mg, 56 μmol, 1.00 eq) under nitrogen atmosphere. The suspension was degassed under vacuum and purged several times with H2. The mixture was stirred at 25 °C under H2 atmosphere for 12 hours. The reaction mixture was filtered and concentrated to give a white solid F1-4 (88 mg, crude).

[0351] Synthesis of compound F1-5

[0352] TFA (2.87 g, 25.2 mmol, 450 eq) was added to a DCM (5 mL) solution of F1-4 (88 mg, 56 μmol, 1.00 eq), and the mixture was stirred at 25 °C for 2 hours. The solution was concentrated, and then 10 mL of DCM and 4 mL of toluene were added. The mixture was evaporated to dryness, and then 0.2 mL of a 4 M HCl solution of dioxane and 10 mL of DCM were added. The mixture was evaporated to dryness, and then 10 mL of DCM and 4 mL of toluene were added again. The mixture was evaporated to dryness to give a white solid product F1-5 (83 mg).

[0353] Synthesis of compound F1

[0354] To a DMF (5.00 mL) solution of F1-5 (60 mg, 43 μmol, 1.00 eq) and Int1 (9 mg, 52 μmol, 1.2 eq, HBr), DIEA (28 mg, 210 μmol, 5.00 eq) and HATU (20 mg, 52 μmol, 1.20 eq) were added. The mixture was stirred at 25 °C for 2 hours. The mixture was quenched with 1 mL of water, and the reaction mixture was purified directly by preparative HPLC (0.075% TFA). The fraction was lyophilized and repurified by preparative HPLC (0.01% HCl) to give F1 (15 mg). LCMS: [M] + =1564.

[0355] Example 7 Synthesis of compounds G1-G4

[0356] Synthesis route:

[0357] Synthesis of compounds G1-G4

[0358] The synthesis of solid and liquid phases is performed according to the methods in Examples 1-6.

[0359] Example 8 HTRF - Experimental Method

[0360] HTRF was used to determine the interaction between PCSK9 and the Alexa Fluor 647 (AF)-labeled cyclic peptide. A solution containing 2 nM biotin-labeled PCSK9 and 5 nM Strep-Eu (Lance streptavidin europium) was prepared in 50 mM HEPES pH 7.4, 0.15 M NaCl, 5 mM CaCl2, 0.01% BSA, and 0.01% surfactant P20. A separate solution containing 120 nM Alexa Flour-labeled cyclic peptide was prepared in the same buffer system. 2 μL of the compound was transferred to the assay plate, followed by the addition of 15 μL of PCSK9 + Strep-Eu and 15 μL of the AF peptide. The final assay volume was 32 μL, containing 1 nM PCSK9, 2.5 nM Strep-Eu, and 56.25 nM AF cyclic peptide. The reaction was incubated at room temperature for 2 hours, followed by fluorescence measurement using an HTRF reader. The IC50 value was determined by fitting the data to an sigmoid dose-response curve using nonlinear regression. Then, the IC50 value was determined based on the K of the AF cyclic peptide. D To calculate Ki.

[0361] Test results: The peptide compounds of this invention exhibit PCSK9 inhibitory activity, wherein the preferred compounds have an IC50 < 50 nm, for example: A1, A2, B1, B4, B6, B7, B8, B12, D1, D2, D3, D4, D5, D6, D7, D8, D9, D10, D11, D12, D13, D14, D15, D16, D17, D18, D19, D21, E1, E2, E8, F1, F2, G3. Wherein:

[0362] Example 9: Pharmacokinetic Study of Compound in Rat

[0363] 1. Experimental Materials

[0364] SD rats: male, 180-250g, purchased from Guangdong Vital River Laboratory Animal Technology Co., Ltd.

[0365] Reagents: DMSO (dimethyl sulfoxide), Labrasol (polyethylene glycol glycerol caprylate), PBS, heparin, acetonitrile, formic acid, and propranolol (internal standard) were all commercially available.

[0366] Instrument: AB SCIEX QTRAP 5500+.

[0367] 2. Experimental Methods

[0368] Weigh out the compound and dissolve it in Labrasol + PBS (30:70, v / v) or 100% PBS. After intravenous or gavage administration to rats, collect 100 μL of venous blood in EDTA-K2 anticoagulant tubes at 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, 24 h, and 28 h (additional 5 min for the IV group). Centrifuge at 12000 rpm for 2 min, and freeze the plasma at -80℃ for later testing. Accurately weigh a certain amount of the test sample and dissolve it in DMSO to a concentration of 2 mg / mL to prepare a stock solution. Accurately pipette an appropriate amount of the compound stock solution and dilute it with acetonitrile to prepare a series of standard solutions. Accurately pipette 10 μL of each of the above standard series solutions, add 90 μL of blank plasma, vortex to mix, and prepare plasma samples with concentrations equivalent to 0.3, 1, 3, 10, 30, 100, 300, 1000, and 3000 ng / mL, and quality control samples with concentrations of 2.4, 120, and 2400 ng / mL. Perform dual-sample analysis for each concentration to establish a standard curve. Take 30 μL of plasma (diluted 5-fold after intravenous administration at 5 min, 15 min, and 30 min), add 150 μL of propranolol (50 ng / mL) in acetonitrile solution, vortex to mix, add 100 μL of purified water, vortex again, centrifuge at 4000 rpm for 5 min, and use the supernatant for LC-MS analysis. The LC-MS detection conditions are as follows:

[0369] Column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0370] Mobile phase: water (0.1% formic acid) - acetonitrile. Gradient elution is performed according to the table below.

[0371] 3. Data Processing

[0372] After LC-MS was used to detect blood drug concentrations, pharmacokinetic parameters were calculated using WinNonlin 6.1 software and a non-compartmental model method.

[0373] Test results: The compounds of this invention exhibit improved absolute bioavailability and / or exposure.

[0374] The results of the preferred compounds are as follows:

[0375] Example 10: Study on the efficacy of B6-hPcsk9 in rats

[0376] 1. Experimental Materials

[0377] 1) Test substance

[0378] ①Medium: 90% PBS+10% Labrasol;

[0379] ② Preparation method of test sample: Accurately weigh the test sample compound, add PBS and stir gently until no particulate matter is present, then add 10% Labrasol to prepare the test sample compound with a final concentration of 0.6 or 3 mg / mL. After aliquoting, store at -20℃ protected from light.

[0380] 2) Basic information about the reagents:

[0381] 3) Information on laboratory animals:

[0382] 2. Grouping and administration:

[0383] 1) After the quarantine and adaptation periods, blood was collected from the orbital venous plexus, and the animals were randomly divided into 3 groups based on baseline LDL-C and TC levels in their serum. Animal grouping information, administration methods, and frequencies are detailed in the group and dosing design section.

[0384] 2) Blood sample collection: After administration, 200 μL of blood was collected from the orbital venous plexus at D5, centrifuged (4℃, 4000 rpm, 10 min) to collect serum, and LDL-C and TC were detected by a fully automated biochemical analyzer;

[0385] 3. Detection indicators:

[0386] 1) Blood lipid test:

[0387] 50 μL of serum was collected, and the following indicators were detected using a fully automated biochemical analyzer: LDL-C and TC.

[0388] IV. Statistical Analysis:

[0389] The results are as follows:

[0390] The compounds of the present invention exhibit good in vivo efficacy with LDL-C and / or TC inhibition rates, preferably with an LDL-C inhibition rate >20% and / or a TC inhibition rate >20%.

[0391] Example 11: Evaluation of drug efficacy in monkeys

[0392] 1. Grouping and Dosing Information

[0393] Animal grouping: Eight male animals were selected for this experiment and randomly assigned to groups (4 males / group) based on their latest body weight and serum LDL-c level. All experimental animals were administered 1.5 mg / kg of the test drug; administration method: oral administration. The administration volume was 2 mL / kg.

[0394] Dosing frequency: All experimental animals were administered the medication once daily for a total of 14 times, from Day 0 to Day 13. The required volume of the formulation solution was calculated based on the animal's body weight and the prescribed dosage before each administration.

[0395] The date of the first administration is recorded as Day 0, and the date of the first gavage is recorded as 0h.

[0396] 2. Blood sample collection and processing

[0397] Blood samples should be collected according to the table below: All animals should be fasted overnight before sampling and administration.

[0398] Blood samples were collected from the cephalic vein or saphenous vein (or other suitable sites) of all experimental animals at the sampling time points, and the actual time of each sample collection was recorded. Samples for blood biochemistry testing were collected into blood collection tubes with separating gel and transferred on wet ice. They were allowed to stand at room temperature for at least 30 min, centrifuged at 4°C (3000×g) for 10 min, and the supernatant serum was separated into labeled EP tubes for immediate use or storage at -80°C. Samples for PK testing were collected into blood collection tubes containing EDTA-K2. Immediately after blood collection, the blood collection tubes were slowly inverted several times, centrifuged at 4°C (3000×g) for 10 min within 60 min, and the supernatant plasma was separated into labeled EP tubes for storage at -80°C.

[0399] The reduction in TC, HDL-c, TG, and ApoB levels from baseline was measured in each treatment group.

[0400] It should be understood that the above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Those skilled in the art can make improvements or modifications based on the above description, and all such improvements and modifications should fall within the protection scope of the appended claims of the present invention.

Claims

1. A PCSK9 cyclic peptide inhibitor, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (I): in, R1 is selected from substituted or unsubstituted phenyl, 5-10-membered heteroaryl, and the substituent is selected from halogen, cyano, C1-C6 alkyl, C1-C6 alkoxy, halo-C1-C6 alkyl, halo-C1-C6 alkoxy, C3-C6 cycloalkyl, C3-C6 heterocycloalkyl, C3-C6 cycloalkylalkyl, C3-C6 cycloalkyloxy, C3-C6 heterocycloalkyloxy, amino, alkyl-substituted amino, aminoalkyl, aminoalkoxy, alkyl-substituted aminoalkyl, or two substituents forming cycloalkyl or heterocycloalkyl; R2, R 3a R 3b R4, R5, R6, R7, and R8 are all independently selected from H, halogens, C1-C6 alkyl groups, halo-C1-C6 alkyl groups, or R7 and R8 are cycloalkyl groups; Y is selected from hydroxyl or amino groups; T1, T2, and T3 are all independently selected from substituted or unsubstituted C1-C6 alkyl, C1-C6 alkyl-benzene-C1-C6 alkyl, C1-C6 alkyl-5-10 heteroaryl-C1-C6 alkyl, and the substituents are selected from halogen, C1-C6 alkyl, and C1-C6 alkoxy. Connect T1 to C or T2; T4 is selected from substituted or unsubstituted C1-C6 alkyl groups, and the substituents are selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, halo-C1-C6 alkyl groups, halo-C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, C3-C6 heterocyclic alkyl groups, C3-C6 cycloalkylalkyl groups, C3-C6 cycloalkyloxy groups, C3-C6 heterocyclic alkyloxy groups, amino groups, alkyl-substituted amino groups, aminoalkyl groups, alkyl-substituted aminoalkyl groups, or two substituents forming cycloalkyl or heterocyclic alkyl groups; Ring A is selected from substituted or unsubstituted phenyl groups, 5-10-membered heteroaryl groups, and the substituents are selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, and halo-C1-C6 alkyl groups. B is selected from -C(O)-NH- or -NH-C(O)-, or B forms a heterocyclic alkyl group with T4, and the heterocyclic alkyl group can be further replaced by hydroxyl, halogen, alkyl, or haloalkyl. C is selected from -C(O)-N- or N; D is selected from Among them, X1, X2, X3, and X4 are all independently selected from CH or N. When it is CH, it can be further substituted by halogens or C1-C6 alkyl groups. X5, X6, X7, and X8 are all independently selected from C, CH, or N. There is a single or double bond between X5 and X6, and a single or double bond between X7 and X8. Alternatively, one of X6 or X7 may not exist. When it is CH, it may be further substituted by halogens or C1-C6 alkyl groups. R9 is selected from alkyl or alkylamino groups, and the alkyl or alkylamino groups may be further replaced by halogens or C1-C6 alkyl groups; m is selected from 1, 2, or 3. R 10 R 11 Each is independently selected from H, halogen, hydroxyl, C1-C6 alkyl, halo-C1-C6 alkyl, or R. 10 R 11 Formation of cycloalkyl groups; R 12 Selected from C1-C6 alkyl groups or not present; Wherein, R2 is not hydrogen; And / or when T4 is CH3-CH, B is not -NH-C(O)-; And / or when R 3a Methyl, R 3b When Y is hydrogen, it is not a hydroxyl group; and / or R1 is not And / or when m is 2, R 10 It is not methyl.

2. The PCSK9 cyclic peptide inhibitor according to claim 1, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (II): Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X6, and m are the same as in claim 1.

3. The PCSK9 cyclic peptide according to claim 1, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (III): Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X7, and m are the same as in claim 1.

4. The PCSK9 cyclic peptide according to claim 1, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (IV): Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X6, and m are the same as in claim 1.

5. The PCSK9 cyclic peptide according to claim 1, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (V): Among them, R1-R 12 The definitions of Y, T1-T4, AC, X1, X3, X7, X8, and m are the same as in claim 1.

6. The PCSK9 cyclic peptide according to claim 1, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The structure of the PCSK9 cyclic peptide inhibitor is shown in general formula (VI): T4 is selected from substituted or unsubstituted C1-C6 alkyl groups, with substituents selected from halogens, C1-C6 alkyl groups, C1-C6 alkoxy groups, halo-C1-C6 alkyl groups, halo-C1-C6 alkoxy groups, C3-C6 cycloalkyl groups, C3-C6 heterocyclic alkyl groups, C3-C6 cycloalkylalkyl groups, C3-C6 cycloalkyloxy groups, C3-C6 heterocyclic alkyloxy groups, amino groups, alkyl-substituted amino groups, aminoalkyl groups, alkyl-substituted aminoalkyl groups, or two substituents forming a cycloalkyl or heterocyclic alkyl group; B is selected from -C(O)-NH-, or -NH-C(O)-, or B forms a heterocyclic alkyl group with T4, and the heterocyclic alkyl group may be further substituted with hydroxyl groups, halogens, alkyl groups, or haloalkyl groups. + It represents a cation that forms a salt with a suitable acid.

7. The PCSK9 cyclic peptide according to claim 6, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The *-T4-B-* is selected from 8. The PCSK9 cyclic peptide inhibitor according to any one of claims 1-7, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, characterized in that, The PCSK9 cyclic peptide inhibitors are selected from: A1-A7, B1-B12, C1-C3, D1-D29, E1-E8, F1-F2, and G1-G4.

9. A pharmaceutical composition, characterized in that, It comprises the PCSK9 cyclic peptide inhibitor according to any one of claims 1-8, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients and / or carriers.

10. Use of the PCSK9 cyclic peptide inhibitor of any one of claims 1-8, or its stereoisomer, racemate, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 9, in the preparation of a medicament for the prevention or treatment of diseases related to the PCSK9 receptor, preferably from the perspective of hyperlipidemia.

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