Class of benzoylglycine derivatives, preparation method therefor , and use thereof

Abstract

Provided are a compound as an LFA-1 antagonist and / or an RASP trapping agent and a stereoisomer thereof, a tautomer thereof, or a mixture thereof, a pharmaceutically acceptable salt thereof, a preparation method for the compound, and use thereof in treating LFA-1 and / or RASP. In-vitro activity test results show that the compound has excellent capabilities of trapping toxic aldehyde and inhibiting the binding of LFA-1 to ICAM-1.

Description

A class of benzoylglycine derivatives, their preparation methods and applications

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority and benefit to Chinese Patent Application No. 202410504456.5, filed with the China National Intellectual Property Administration (CNIPA) on April 25, 2024; Chinese Patent Application No. 202411248025.3, filed with the CNIPA on September 6, 2024; and Chinese Patent Application No. 202411679367.0, filed with the CNIPA on November 22, 2024, the disclosures of which are incorporated herein by reference in their entirety. Technical Field

[0003] This application relates to the field of pharmaceutical technology, and more specifically, to a class of benzoylglycine derivatives and methods for preparing and using the compounds. Background Technology

[0004] Dry eye syndrome, also known as keratoconjunctivitis sicca, is a difficult-to-treat ocular surface disease. Its main symptoms include dryness, a foreign body sensation, and burning. Dry eye syndrome is a chronic ocular surface disease caused by multiple factors. The medical community generally believes that its main pathological mechanism is tear film instability or ocular surface microenvironment imbalance due to abnormalities in the quality or quantity of tears, accompanied by ocular surface inflammation, tissue damage, and abnormal nerve sensation. If dry eye syndrome is not diagnosed and treated promptly, it may lead to further complications such as infection, ocular surface keratinization, corneal ulceration, and conjunctival squamous metaplasia.

[0005] Dry eye syndrome is a continuous pathological process, progressing from mild to severe without clear boundaries between mild, moderate, and severe stages. Currently, treatment for dry eye syndrome mainly falls into three categories: medication, physical therapy, and surgery.

[0006] Although the etiology of dry eye is complex, researchers have found similar pathologies: immune cells infiltrate the surface tissues of the eye, causing chronic inflammation and resulting in ocular surface damage. Anti-inflammatory treatment has become the direction for the development of drugs for dry eye. Currently, there are two main anti-inflammatory directions in the European and American markets: immunosuppressants and active aldehyde scavengers. Two immunosuppressant drugs have been approved: (1) Cyclosporine A suspension. This drug is a suspension, which has problems such as instability during long-term storage and eye irritation from the suspension. In addition, as a strong immunosuppressant, it may cause damage to the immune system. (2) Liftitigrast, an LFA-1 antagonist, can inhibit the binding of lymphocyte function-associated antigen 1 (LFA-1) to intercellular adhesion molecule 1 (ICAM-1), thereby reducing the level of inflammation mediated by T lymphocytes to exert its effect. It was approved by the FDA in December 2016. However, it has high lipophilicity, and long-term use can lead to taste degeneration. Moreover, it has no effect on more than 50% of patients in clinical practice. The most advanced active aldehyde scavenger is ADX-102. RASP (active aldehyde) undergoes a covalent reaction with amino and thiol groups. After binding to a threshold amount in the electrophilic response proteome, RASP affects protein function, such as MAP kinase, protein kinase C, and other proteins that enhance cytokine release and inflammation, and activates intracellular inflammatory factors, such as NF-κB. Furthermore, RASP protein adducts bind to scavenger receptor A, which initiates pro-inflammatory signaling and forms antibodies against the adduct protein, thereby enhancing the inflammatory response. ADX-102 can capture RASP, thereby blocking its mediated subsequent series of reactions, resulting in an anti-inflammatory effect and thus achieving the goal of treating dry eye syndrome. However, ADX-102 has poor water solubility; even with the help of excipients, the highest concentration of eye drops is only 0.5%. Due to insufficient efficacy and excessive ocular irritation, supplementary phase III clinical trials are currently underway. Summary of the Invention

[0007] The purpose of this application is to provide a novel compound as an LFA-1 antagonist and / or RASP scavenger, a method for preparing the compound, and its use in treating eye diseases (e.g., dry eye syndrome).

[0008] A first aspect of this application provides a compound of the following formula (Va), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0009] in,

[0010] Ring A is a 5-12 membered heteroaryl or a C6-12 aryl;

[0011] Ring B is a 5-12 membered heteroaryl or a C6-12 aryl;

[0012] R A Each of the following is independently selected from deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more elements selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups;

[0013] R B Each of the following groups is independently selected from deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups;

[0014] a is selected from 0, 1, 2, 3, 4, 5, or 6;

[0015] b is selected from 1, 2, 3, 4, 5, or 6;

[0016] R S Each element is independently selected from deuterium, halogens, -CN, -OH, -NH2, and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkyl thio group; or 2 R groups S Together with the atoms it is attached to, they form C 3-8 cycloalkyl;

[0017] n is 0, 1, 2, 3 or 4;

[0018] R is independently selected from hydrogen, halogen, or C. 1-6 alkyl;

[0019] Unless otherwise stated, the heteroatoms in the above heteroaryl groups are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4;

[0020] The condition is that, In this case, 'a' must be at least 2, and there are two 'R's. A They are -NH2 and And adjacent, and / or, In the case of b, it is at least 2, and there are two R values. B They are -NH2 and And they are adjacent.

[0021] This application also provides a compound represented by the formula (V-a'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0022] Among them, rings A and R A Ring B, R B R S a, b, n, R are as described in compound (Va).

[0023] This application also provides a compound represented by formula (I'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0024] in,

[0025] Ring A is a 5-12 membered heteroaryl or a C6-12 aryl;

[0026] Ring B is a 5-12 membered heteroaryl or a C6-12 aryl;

[0027] R A For deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more elements selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups;

[0028] R B For deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups;

[0029] a is selected from 0, 1, 2, 3, 4, 5, or 6;

[0030] b is selected from 1, 2, 3, 4, 5, or 6;

[0031] R is independently selected from hydrogen, halogen, or C. 1-6 alkyl;

[0032] Unless otherwise stated, the heteroatoms in the above heteroaryl groups are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4;

[0033] The condition is that, Include Structure, and / or Include Structure; dashed lines represent single or double bonds.

[0034] In equation (Va), equation (V-a'), or equation (I'):

[0035] In some implementations, ring A is a 6-10 membered heteroaryl or phenyl group.

[0036] In some embodiments, ring A is a 6-10-membered heteroaryl or phenyl group, wherein the 6-10-membered heteroaryl group contains one or two heteroatoms independently selected from N or O.

[0037] In some embodiments, ring A is pyridyl, quinolinyl, or phenyl.

[0038] In some embodiments, ring A is pyridyl, quinolinyl, phenyl, or benzofuranyl.

[0039] In some implementation schemes, R A Each of the following is independently selected from deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C1-6 alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Alkyl groups are substituted.

[0040] In some implementation schemes, R A C is independently selected from halogens, -OH, -NH2, or optionally substituted: 1-6 Alkyl, C3-6 cycloalkyl, C 1-6 Alkoxy; wherein optional substitution means either unsubstituted or substituted by one or more substituents selected from halogens and -OH.

[0041] In some implementation schemes, R A -NH2 or

[0042] In some implementations, 'a' is selected from 1, 2, 3, 4, 5, or 6.

[0043] In some implementations, 'a' is selected from 0, 1, 2, or 3.

[0044] In some implementations, 'a' is selected from 0, 2, 3, or 4.

[0045] In some implementations, 'a' is selected from 1 or 2.

[0046] In some implementations, 'a' is selected from 2 or 3.

[0047] In some implementations, a is 2.

[0048] In some implementations, a is 2, and both R A Located in the adjacent position.

[0049] In some implementations, a is 2, and both R A Located adjacent, two R A They are -NH2 and

[0050] In some implementations, a is 3, and two of the R values ​​are 3. A Located adjacent, two R A They are -NH2 and Another R A Selected from deuterium, halogens, -CN, -OH, -NH2, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group.

[0051] In some implementations, a is 3, and two of the R values ​​are 3. A Located adjacent, two R A They are -NH2 and Another R A Selected from deuterium, halogen, -CN, -OH, -NH2, methyl, cyclopropyl or trifluoromethyl.

[0052] In some implementations, a is 3, and two of the R values ​​are 3. A Located adjacent, two R A They are -NH2 and Another R A Selected from F, Cl, -OH, methyl, methoxy, cyclopropyl or trifluoromethyl.

[0053] In some implementations, a is 4, and two of them R A Located adjacent, two R A They are -NH2 and The other two R A Selected from halogens (e.g., F).

[0054] In some embodiments, for X1, X2, or X3 are each independently selected from CR A Or N.

[0055] In some embodiments, for X1, X2, or X3 are each independently selected from CH and CR. A Or N.

[0056] In some embodiments, for X1, X2, or X3 are each independently selected from CH and CR. A Or N.

[0057] Each R A Independently deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more elements selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 The alkyl group is substituted. In some embodiments, each R... A Independently, it is deuterium, halogen, -CN, -OH, -NH2, methyl, trifluoromethyl, methoxy, or cyclopropyl. In some embodiments, each R... A Independently, it can be deuterium, halogen, -CN, -OH, or -NH2. In some embodiments, each R... A It is F.

[0058] In some embodiments, for X1, X2, or X3 are each independently selected from CH and CR. A Or N; or one of X1, X2, or X3 is N, and the other two are CH; or one of X1, X2, or X3 is N, and the other two are CH and CR respectively. A Alternatively, one of X1, X2, or X3 is N, and the other two are CR. A ; or X1, X2, or X3 are all CH; or one of X1, X2, or X3 is CR. A The other two are both CH; or one of X1, X2, or X3 is CH, and the other two are both CR. A .

[0059] Each R A Independently deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted with one or more radicals selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 The alkyl group is substituted. In some embodiments, each R... A Independently halogenated, -OH, or optionally substituted: C 1-6 Alkyl, C 3-6 cycloalkyl, C 1-6 Alkyl groups; wherein optional substitution means either unsubstituted or substituted with one or more halogens. In some embodiments, each R A Independently, it is F, Cl, -OH, methyl, trifluoromethyl, methoxy, or cyclopropyl. In some embodiments, each R... A It is F.

[0060] In some embodiments, for

[0061] In some embodiments, for

[0062] In some embodiments, for

[0063] In some embodiments, ring B is a 6-10-membered heteroaryl or phenyl group. In some embodiments, ring B is a 6-10-membered heteroaryl or phenyl group, wherein the 6-10-membered heteroaryl group contains one or two heteroatoms independently selected from N or O.

[0064] In some embodiments, ring B is pyridyl or phenyl.

[0065] In some implementation schemes, R B -NH2, methanesulfonyl or

[0066] In some implementations, b is selected from 1 or 2.

[0067] In some implementations, when b is 1, R B It is methanesulfonyl; when b is 2, R B They are -NH2 and

[0068] In some implementations, when b is 1, R B It is methanesulfonyl; when b is 2, R B They are -NH2 and And two Rs B Adjacent.

[0069] In some embodiments, for

[0070] In some embodiments, for

[0071] In some implementation schemes, R S Selected independently from deuterium, halogens, and C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 cycloalkyl or C 1-4 alkoxy group; or 2 R groups S Together with the atoms it is attached to, they form C 3-6 Cycloalkyl.

[0072] In some implementation schemes, R S Selected independently from deuterium, halogens, and C 1-4 Alkyl or C 1-4 Alkoxy.

[0073] In some implementation schemes, R S Each of the following is independently selected from deuterium, F, methyl, ethyl, methoxy; or two Rs. S Together with the atoms it is attached to, it forms a cyclopropyl group.

[0074] In some implementations, n is 0, 1, or 2.

[0075] In some implementations, n is 0 or 1.

[0076] In some implementations, n is 0.

[0077] In some implementations, n is 1 or 2, R S It is located at the meta position of the N atom on the ring.

[0078] In some implementations, n is 1, R S It is located in the adjacent position of the N atom on the ring.

[0079] In some implementations, R is independently selected from hydrogen or halogen.

[0080] In some implementations, R is independently selected from hydrogen or Cl.

[0081] In some implementations, the two Rs are hydrogen and Cl, respectively.

[0082] In some implementations, both R are Cl.

[0083] In some embodiments, ring A is pyridyl or phenyl, a is 2, and both R... A Located adjacent, two R A They are -NH2 and for n is 0.

[0084] In some embodiments, ring A is phenyl, a is 3 or 4, and two of the Rs are phenyl. A Located adjacent, two R A They are -NH2 and Another or two R A Selected from F or Cl, n is 0, or n is 1, R S It can be methyl or ethyl.

[0085] This application also provides a compound represented by formula (II'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0086] Among them, rings A and R A a, R as described in compounds of formula (Va) or formula (I').

[0087] This application also provides a compound represented by formula (III'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0088] Among them, rings A, B, R, and R B b, as described in compounds of formula (Va) or (I').

[0089] This application also provides a compound represented by formula (I), formula (II) or formula (III) as follows, a stereoisomer, tautomer or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0090] Among them, rings A and R A Ring B, R B a, b are as described in compound (I');

[0091] Among them, rings A and R A a is as described in compound of formula (I');

[0092] Among them, ring A, ring B, R B b is as described in compound (I').

[0093] This application also provides a compound of formula (V), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0094] Among them, ring A, ring B, R A R B R S a, b, and n are as described in equation (Va).

[0095] This application also provides a compound represented by formula (VI), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0096] Among them, X1, X2, or X3 are independently selected from CH and CR, respectively. A Or N; ring B, R A R B b, R, R S As described in compound (Va).

[0097] This application also provides a compound represented by formula (VII), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0098] Among them, rings B and R A R B b, R, R S As described in compound (Va).

[0099] This application also provides a compound represented by formula (VI-a) as follows, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0100] Among them, X1, X2, or X3 are independently selected from CH and CR, respectively. A Or N; R A R S As described in compound (Va).

[0101] This application also provides a compound represented by formula (VII-a), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0102] where R A R S As described in compound (Va).

[0103] In formulas (I), (II), (III), (I'), (II'), (III'), (V), (Va), (VI), (VI-a), (VII), and (VII-a), the C atom bonded to COOH has the (S) configuration.

[0104] In some implementations, this application includes the variables defined above and their implementations, as well as any combination thereof.

[0105] The compounds described in this application, stereoisomers, tautomers, or mixtures thereof, or pharmaceutically acceptable salts of the compounds, are selected from:

[0106] In a second aspect, this application provides pharmaceutical compositions comprising the compounds described in the first aspect, their stereoisomers, tautomers, or mixtures thereof, or pharmaceutically acceptable salts, and optionally, further comprising a pharmaceutically acceptable carrier.

[0107] In some embodiments, the pharmaceutical composition comprises a compound as described in the first aspect, a stereoisomer thereof, a tautomer thereof, or a mixture thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0108] In some embodiments, the pharmaceutical composition serves as an LFA-1 antagonist and / or a RASP trapping agent.

[0109] In some embodiments, the pharmaceutical composition is used to treat and / or prevent LFA-1-mediated diseases and / or diseases involving aldehyde toxicity, comprising a compound as described in the first aspect, its stereoisomers, tautomers or mixtures thereof, isotopic derivatives or pharmaceutically acceptable salts, and one or more pharmaceutically acceptable carriers.

[0110] In some embodiments, the pharmaceutical composition is used to treat and / or prevent LFA-1-mediated diseases and / or diseases involving aldehyde toxicity, comprising a therapeutically effective amount of the compound as described in the first aspect, its stereoisomers, tautomers or mixtures thereof, isotopic derivatives or pharmaceutically acceptable salts, and one or more pharmaceutically acceptable carriers.

[0111] Thirdly, this application provides the use of the compounds, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect, in the preparation of a medicament.

[0112] In some embodiments, the drug is a drug that acts as an LFA-1 antagonist and / or a RASP scavenger.

[0113] In some embodiments, the drug is a drug for treating and / or preventing LFA-1-mediated diseases and / or diseases whose pathogenesis involves aldehyde toxicity.

[0114] Fourthly, this application provides compounds, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect, for the treatment and / or prevention of LFA-1-mediated and / or pathogenesis involving aldehyde toxicity.

[0115] Fifthly, this application provides methods for treating and / or preventing LFA-1-mediated diseases and / or diseases involving aldehyde toxicity, comprising administering to a subject a therapeutically effective amount of the compound, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect, the application of ...

[0116] Sixthly, this application provides the use of the compounds, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts, or pharmaceutical compositions as described in the first aspect, in the treatment and / or prevention of LFA-1-mediated and / or pathogenesis involving aldehyde toxicity.

[0117] In the second, third, fourth, fifth, and sixth aspects, diseases mediated by LFA-1 and / or whose pathogenesis involves aldehyde toxicity are preferably ocular diseases; preferably, the ocular disease is a chronic ocular surface disease; preferably, the ocular disease is dry eye syndrome.

[0118] In the absence of contradictions and conflicts, the technical solutions or features described in the third aspect are applicable to the fourth, fifth and sixth aspects.

[0119] The compounds or pharmaceutical compositions of this application have one or more of the following beneficial effects: (1) the compounds of this application have the ability to capture toxic aldehydes; (2) the compounds of this application can effectively inhibit the binding of LFA-1 and ICAM-1; (3) the compounds of this application have the effect of inhibiting cell adhesion.

[0120] The purpose of this application also includes providing methods for preparing the compounds shown above, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of said compounds.

[0121] Including but not limited to the following methods:

[0122] General preparation method one:

[0123] Among them, ring A, ring B, R A a, R B b is as described in formula (I') or the compound of formula (I).

[0124] General preparation method two:

[0125] Among them, rings A and R A a is as described in compounds of formula (I') or (II).

[0126] General preparation method three:

[0127] Among them, rings B and R B R S X1, X2, X3, b, n are as described in compounds of formula (VI) or (VI-a).

[0128] General preparation method four:

[0129] Among them, rings B and R B R S X1, X2, X3, b, n are as described in compounds of formula (VI) or (VI-a), where HG is a carboxyl protecting group.

[0130] The purpose of this application also includes providing intermediates for preparing the compounds shown above, stereoisomers, tautomers or mixtures thereof, or pharmaceutically acceptable salts of the compounds: 1-c, 1-d, 1-e, 1-g, 2-g, 3-a, 3-b, 3-d, 3-e, 3-g, 4-c, 4-d, 4-e.

[0131] definition

[0132] Unless otherwise stated, the following terms as used in this application have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0133] Unless otherwise specified, the term "alkyl" refers to a monovalent saturated aliphatic hydrocarbon group, a straight-chain or branched group comprising 1-20 carbon atoms, preferably comprising 1-10 carbon atoms (i.e., C10). 1-10 Alkyl groups, more preferably containing 1-8 carbon atoms (C64- ... 1-8 Alkyl groups, more preferably containing 1-6 carbon atoms (i.e., C64-C ... 1-6 Alkyl groups, more preferably containing 1-4 carbon atoms (i.e., C464-C ... 1-4 Alkyl groups, more preferably containing 1-3 carbon atoms (i.e., C464-C ... 1-3 Alkyl), for example, "C 1-6 "Alkyl" refers to a group that is alkyl and has 1 to 6 carbon atoms in its carbon chain (specifically, 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, neopentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, n-heptyl, n-octyl, etc.

[0134] The substitution of "alkyl" with halogen refers to "halogenated alkyl", such as C 1-6 Alkyl substitution by halogens refers to C 1-6 Haloalkyl. "Haloalkyl" can be C10 or C20. 1-6 Halogenated alkyl or C 1-4 Halogenated alkyl groups.

[0135] Unless otherwise specified, the terms "halogen" or "halogenated" refer to F, Cl, Br, and I. The term "halogenated alkyl" refers to an alkyl group as defined above in which one, two, or more hydrogen atoms, or all hydrogen atoms, are replaced by a halogen. Representative examples of halogenated alkyl groups include CCl3, CF3, CHCl2, CH2Cl, CH2Br, CH2I, CH2CF3, and CF2CF3.

[0136] Unless otherwise specified, the term "cycloalkyl" refers to a monocyclic saturated aliphatic hydrocarbon group having a specific number of carbon atoms, preferably containing 3-12 carbon atoms (i.e., C12-C12). 3-12 cycloalkyl), more preferably containing 3-10 carbon atoms (C 3-10 cycloalkyl groups, more preferably 3-6 carbon atoms (C 3-6 cycloalkyl groups), 4-6 carbon atoms (C 4-6 cycloalkyl groups), 5-6 carbon atoms (C 5-6 (Cycloalkyl). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, etc.

[0137] Unless otherwise specified, the term "alkoxy" refers to an alkyl residue in which one or more carbon atoms (and associated hydrogen atoms) are replaced by oxygen, such as "alkoxy" or "alkoxyalkyl". Examples include methoxy, ethoxy, propoxy, methoxypropyl, etc. "Alkoxy" can be C10, C20, C30, C40, C50, C60, C7 ... 1-6 Alkoxy or C 1-4 Alkoxy.

[0138] Unless otherwise specified, the term "alkoxy" refers to a group in which the oxygen in "alkoxy group" is replaced by sulfur or nitrogen. "Alkoxy" can be C10 or C20. 1-6 alkylthio or C 1-4 Alkylthio group.

[0139] Unless otherwise specified, the term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic cyclic hydrocarbon substituent, which is a non-aromatic structure containing 3-20 ring atoms, wherein one, two, three, or more ring atoms are selected from N, O, or S, and the remaining ring atoms are C. Preferably, it contains 3-12 ring atoms, more preferably 3-10 ring atoms, or 3-8 ring atoms, or 3-6 ring atoms, or 4-6 ring atoms, or 5-6 ring atoms. The number of heteroatoms is preferably 1-4, more preferably 1-3 (i.e., 1, 2, or 3). Examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazoalkyl, tetrahydrofuranyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, pyranyl, etc. Bicyclic or polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged-ring heterocyclic groups.

[0140] Unless otherwise specified, the term "aryl" refers to an aromatic carbocyclic system containing 6-16 carbon atoms, or 6-14 carbon atoms, or 6-12 carbon atoms, or 6-10 carbon atoms, preferably 6-10 carbon atoms. The term "aryl" may be used interchangeably with the term "aromatic ring." Examples of aryl groups may include, but are not limited to, phenyl, naphthyl, anthraceneyl, phenanthryl, or pyreneyl.

[0141] Unless otherwise specified, the term "heteroaryl" refers to an aromatic monocyclic, bicyclic, or polycyclic cyclic system containing a 5-16 member structure, or a 5-14 member structure, a 5-12 member structure, a 5-10 member structure, a 6-10 member structure, a 5-8 member structure, or a 5-6 member structure, wherein one, two, three, or more ring atoms are heteroatoms and the remaining atoms are carbon atoms, the heteroatoms being independently selected from O, N, or S, and the number of heteroatoms is preferably one, two, or three. Examples of heteroaryl groups may include, but are not limited to, furanyl, thiophene, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiazolyl, pyrrole, pyrazolyl, imidazole, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiodiazolyl, triazinyl, phthalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purine, indoleyl, isoindoleyl, indazoleyl, benzofuranyl, benzothiophene, benzopyridyl, benzopyrimidinyl, and benzene. Pyrazinyl, benzimidazolyl, benziphthalazolyl, pyrrolo[2,3-b]pyridyl, imidazo[1,2-a]pyridyl, pyrazolo[1,5-a]pyridyl, pyrazolo[1,5-a]pyrimidinyl, imidazo[1,2-b]pyridazinyl, [1,2,4]triazolo[4,3-b]pyridazinyl, [1,2,4]triazolo[1,5-a]pyrimidinyl, [1,2,4]triazolo[1,5-a]pyridyl, etc.

[0142] The contents recorded in this article This indicates that ring A contains at least NH2 and -C(CH3)2OH substituents, and that they are in adjacent positions on ring A. The contents described herein... This indicates that ring B contains at least NH2 and -C(CH3)2OH substituents, and they are in adjacent positions on ring B. Other substituents are similar.

[0143] The contents recorded in this article This indicates that the relative positions of the carbonyl group with NH2 and / or -C(CH3)2OH on ring A are variable. It's similar.

[0144] Unless otherwise specified, the terms "pharmaceutically acceptable salt" or "medicinal salt" refer to a salt that, within the bounds of reasonable medical judgment, is suitable for contact with the tissues of mammals, particularly humans, without excessive toxicity, irritation, allergic reactions, etc., and is proportionate to a reasonable benefit / risk ratio. Medically acceptable salts of amines, carboxylic acids, and other types of compounds are well known in the art. The salt can be prepared in situ during the final isolation and purification of the compounds of this application, or solely by reacting a free base or free acid with a suitable reagent.

[0145] The compounds in this application also include their isotopic derivatives. Unless otherwise specified, the term "isotopic derivative" refers to compounds in this application that can exist in an isotopically traced or enriched form, containing one or more atoms whose atomic weights or mass numbers differ from the atomic weights or mass numbers of the most abundant atoms found in nature. Isotopes can be radioactive or non-radioactive. Commonly used isotopes for isotopic labeling are: hydrogen isotopes, 2 H and 3 H; Carbon isotopes: 13 C and 14 C; Chlorine isotopes: 35 Cl and 37 Cl; Fluorine isotopes: 18 F; Iodine isotopes: 123 I and 125 I; Nitrogen isotopes: 13 N and 15 N; oxygen isotopes: 15 O, 17 O and 18 O and sulfur isotopes 35 S. These isotope-labeled compounds can be used to study the distribution of pharmaceutical molecules in tissues. Especially 3 H and 13 C, because they are easy to label and convenient to detect, are more widely used. Some heavy isotopes, such as deuterium (… 2 Substitution with H can enhance metabolic stability and prolong the half-life, thereby achieving the goal of reducing dosage and providing therapeutic advantages. Isotope-labeled compounds are generally synthesized from labeled starting materials using known synthetic techniques, just like non-isotope-labeled compounds.

[0146] The compounds in this application also include their solvates or solvent compounds. Unless otherwise specified, the terms "solvate" or "solvent compound" refer to the physical association of the compound with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. Solvent molecules in a solvate may be present in a regular and / or disordered arrangement. Solvates may contain stoichiometric or non-stoichiometric solvent molecules. "Solvate" encompasses both solution phases and separable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Solvation methods are well known in the art.

[0147] Unless otherwise specified, the term "stereoisomer" refers to compounds having the same chemical structure but with different spatial arrangements of atoms or groups. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and inhibited isomers. Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0148] Unless otherwise specified, the term "tautomer" refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium can be achieved in the tautomer. For example, proton tautomers (also called proton transfer tautomers) involve interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers involve interconversions via the rearrangement of some bonding electrons.

[0149] Unless otherwise indicated, the structural formulas described in this application include all isomers (such as enantiomers, diastereomers, and geometric isomers (or conformational isomers)): for example, R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, any single stereochemical isomer of the compound of this application, or its enantiomers, diastereomers, or mixtures of geometric isomers (or conformational isomers), are within the scope of this application.

[0150] The compounds in this application also include their cocrystals. Unless otherwise specified, the term "cocrystal" is used to describe a situation in which neutral molecular components are present in a definite stoichiometric ratio within a crystalline compound. The preparation of pharmaceutical cocrystals allows for alteration of the crystal form of the active pharmaceutical ingredient, which in turn alters its physicochemical properties without impairing its desired biological activity (see Pharmaceutical Salts and Cocrystals, eds. J. Wouters and L. Quere, RSC Publishing, 2012).

[0151] The compounds in this application also include their polymorphs. Unless otherwise specified, the term "polymorph" refers to different arrangements of chemical drug molecules, generally manifested as the form in which the drug raw material exists in a solid state. A drug can exist in multiple crystalline states. Different crystalline forms of the same drug may have different solubility and absorption in the body, thus affecting the dissolution and release of the formulation.

[0152] The compounds in this application also include their metabolites. Unless otherwise specified, the term "metabolite" refers to a product obtained in vivo by the metabolic processes of a specific compound or its salt. Metabolites of a compound can be identified using techniques known in the art, and their activity can be characterized by experimental methods as described in this application. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, acylation, deacylation, esterification, defatting, enzymatic cleavage, etc. Accordingly, this application includes metabolites of compounds, including metabolites produced by sufficient contact of the compounds of this application with mammals for a period of time.

[0153] The compounds in this application also include their prodrugs. Unless otherwise specified, the term "prodrug" refers to a drug that is converted into the parent drug in vivo. Prodrugs are generally useful because they can improve certain, undesirable physical or biological properties. Physical properties are generally related to solubility (excessive or insufficient lipid or water solubility) or stability, while problematic biological properties include metabolism that is too rapid or poor bioavailability, which may itself be related to physicochemical properties. For example, they can be bioavailable orally, whereas the parent drug cannot. Prodrugs also have improved solubility in pharmaceutical compositions compared to the parent drug. An example of a prodrug, but not limited to, is any compound of this application that is administered as an ester ("prodrug") to facilitate transmembrane transport, where water solubility is detrimental to migration but beneficial once inside the cell, and is subsequently metabolized and hydrolyzed into a carboxylic acid, i.e., the active entity. Another example of a prodrug can be a short peptide (polyamino acid) bound to an acid group, where the peptide is metabolized to exhibit the active moiety.

[0154] Unless otherwise specified, the term "optional substitution" means that the hydrogen at the substituted site of the group is not substituted, or is substituted by one or more substituents, preferably selected from the group consisting of: halogen, hydroxyl, mercapto, cyano, nitro, amino, azide, oxo, carboxyl, C 2-6 Alkenyl, C 2-6 alkynyl group, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 cycloalkyl, C 3- 10 Cycloalkylsulfonyl, 3-10 membered heterocyclic alkyl, C 6-14 Aryl or 5-10 membered heteroaryl rings, wherein the C 2-6 Alkenyl, C 2-6 alkynyl group, C 1-6 Alkyl, C 1-6 Alkoxy, C 3-10 cycloalkyl, C 3-10 Cycloalkylsulfonyl, 3-10 membered heterocyclic alkyl, C 6-14The aryl or 5-10 membered heteroaryl group may optionally be selected from halogen, hydroxyl, amino, cyano, C 1-6 Alkyl or C 1-6 One or more substituents in the alkoxy group are substituted, wherein the oxo group refers to two H atoms at the same substitution position being replaced by the same O atom to form a double bond.

[0155] The terms “optional” or “optional” mean that the event or situation described below may, but does not have to, occur, including in cases where the event or situation does not occur.

[0156] The word “comprise” or “include” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0157] The term "subject," also referred to as "individual" or "subject," means a cell or animal, including but not limited to mammals such as laboratory animals or humans. "Effective amount" means an amount of the compound or pharmaceutical composition described herein sufficient to achieve the intended application, including but not limited to the treatment of a disease or the relief of its symptoms. In some embodiments, for example, the amount may be a dose that can induce a specific response in cells, or a dose that exerts a therapeutic effect on a disease in a model animal. Specificity will vary depending on, for example, the specific compound selected, the type of subject and their age / pre-existing health condition, the dosing regimen followed, the severity of the disease, whether it is administered in combination with other agents, the timing of administration, the tissue of administration, and the physical delivery system carrying it.

[0158] Some of the compounds in this application are optically active. The compounds in this application can be racemic, optical isomers or mixtures thereof. The optical isomers in the compounds in this application can be synthesized either by using the starting materials of the optical isomers or by separating the racemic mixtures.

[0159] Unless otherwise specified, the term "pharmaceuticalally acceptable carrier," also known as "pharmaceuticalally acceptable excipient" or "pharmaceuticalally acceptable excipient," refers to a generally recognized medium in the field of delivering bioactive agents to animals (specifically, mammals). Pharmaceutically acceptable carriers are formulated based on a number of factors well understood by those skilled in the art. These include (but are not limited to) the type and nature of the formulated active agent, the subject to whom the composition containing the pharmaceutical agent is to be administered, the intended route of administration of the composition, and the targeted therapeutic indication. Pharmaceutically acceptable carriers include aqueous and non-aqueous liquid media, as well as various solid and semi-solid dosage forms. The preparation of pharmaceutical compositions described herein includes, but is not limited to, mixing, for example, the compounds described in the first or second aspect, or their tautomers, stereoisomers, or pharmaceutically acceptable salts thereof, with a pharmaceutically acceptable carrier.

[0160] The pharmaceutical compositions of this application can be prepared by combining the compounds of this application with suitable pharmaceutically acceptable excipients.

[0161] Typical routes of administration for the compounds of this application or their pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0162] The pharmaceutical composition of this application can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, emulsification, freeze drying, etc.

[0163] In all methods of administration of the compounds of this application, the daily dose may be from 0.5 mg to 20 mg per person, in the form of single or separate doses.

[0164] The above embodiments represent exemplary embodiments of this application, but this application is not limited to the above embodiments. Furthermore, the various technical features in the above embodiments of this application can be combined with each other to constitute one or more new technical solutions, which also fall within the scope of this application, provided that such new technical solutions are technically feasible. Detailed Implementation

[0165] The present application is further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or as recommended by the manufacturer. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this application. The preferred embodiments and materials shown herein are for illustrative purposes only.

[0166] The structures of the compounds in this application were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS) and / or high-performance liquid chromatography (HPLC). The NMR measurements were performed using a Bruker 400MHz and / or Varian 400MHz instrument; the LC-MS instrument used was an Agilent 1260 Infinity II-6120 / 6125MSD; and the HPLC instrument used was a Waters Acquity UPLC_2 and / or Shimadzu LC2030 and / or Agilent 1260 Infinity II. Chiral compound separation was performed using an SFC-150 (Waters) instrument and a DAICEL chiral column. Column volume: 20×250mm (10μm particle size packing).

[0167] The starting materials used in the embodiments of this application are known and commercially available, or can be synthesized using methods known in the art.

[0168] This application provides a method for preparing the compound. The compound can be prepared by the following steps.

[0169] Preparation Example 1

[0170] Preparation of 3-{[(tert-butoxy)carbonyl]amino}-4-(2-hydroxypropyl-2-yl)benzoic acid (P-4):

[0171] Step 1: P-1 (6.00 g) was dissolved in tetrahydrofuran (200 mL), and a tetrahydrofuran solution of methyl magnesium bromide (3 M, 52 mL) was added. The reaction was carried out at 0°C for 0.5 hours under nitrogen protection. After the reaction was complete as detected by TLC, the reaction solution was quenched with water (100 mL), extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by rapid chromatography (Silica gel, petroleum ether:ethyl acetate = 5:1) to obtain P-2 (5.50 g). LCMS (ESI) [M+H-18] + =211.9.

[0172] Step 2: P-2 (2.50 g) was dissolved in ethanol (50 mL), and di-tert-butyl dicarbonate (3.41 g) was added. The mixture was reacted at 40 °C for 16 hours. The reaction solution was concentrated and purified by rapid chromatography (Silica gel, petroleum ether:ethyl acetate = 10:1) to obtain P-3 (3.00 g). LCMS (ESI) [M+H-Boc-18] + =258.0; 1H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 8.19 (d, J = 1.6Hz, 1H), 7.19–7.09 (m, 2H), 6.17 (s, 1H), 1.49 (s, 6H), 1.47 (s, 9H).

[0173] Step 3: P-3 (3.00 g) was dissolved in dimethyl sulfoxide (30 mL) and water (15 mL). Triethylamine (6.38 mL) and palladium dichloride bis((cyclopentadien-1-yl)diphenylphosphine)iron (999.33 mg) were added. The mixture was stirred at 80 °C for 16 hours under a carbon monoxide (balloon) atmosphere. Water (25 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate. The product in the aqueous phase was adjusted to pH 5 with citric acid (3 M) and extracted with ethyl acetate (25 mL, 3 times). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. P-4 (584.99 mg) was obtained by rapid chromatography (Silica gel, tetrahydrofuran:petroleum ether = 1:5). LCMS (ESI) [M+H-18-t-Bu] + =222.1; 1 H NMR (400MHz, DMSO-d6) δ12.88(s,1H),9.80(s,1H),8.58(d,J=1.5Hz,1H),7.53(dd ,J=8.2,1.8Hz,1H),7.35(d,J=8.2Hz,1H),6.19(s,1H),1.53(s,6H),1.48(s,9H).

[0174] Preparation Example 2

[0175] Preparation of 4-{[(tert-butoxy)carbonyl]amino}-3-(2-hydroxypropane-2-yl)benzoic acid (P-8):

[0176] Step 1: P-5 (6.00 g) was dissolved in tetrahydrofuran (200 mL), and methylmagnesium bromide (43.47 mL) was added dropwise at 0 °C. The reaction was carried out under nitrogen protection at 0 °C for 0.5 hours. After the reaction was complete, water (100 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and then subjected to column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain P-6 (5.1 g). LCMS (ESI) [M+H-18] + =212.1; 1 H NMR (400MHz, DMSO-d6) δ7.11–7.01(m,2H),6.57(d,J=8.4Hz,1H),5.55(s,2H),5.30(s,1H),1.47(s,6H).

[0177] Step 2: P-6 (2.00 g) was dissolved in ethanol (20 mL), and di-tert-butyl dicarbonate (2.85 g) was added. The reaction was carried out at 30 °C for 16 hours. After the reaction was complete, the reaction solution was directly concentrated and purified by column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain P-7 (2.30 g). LCMS (ESI) [M+H-56-23] + =297.3; 1 H NMR (400MHz, DMSO-d6) δ9.68(s,1H),7.90(d,J=8.7Hz,1H),7.42–7.36(m,1H),7.34(d,J=2.3Hz,1H),6.15(s,1H),1.50(s,6H),1.46(s,9H).

[0178] Step 3: Dissolve P-7 (2.20 g) in dimethyl sulfoxide (20 mL) and water (10 mL), then add triethylamine (6.74 mL) and palladium (2 g). + 0.98 g of bis((cyclopenten-1,3-dien-1-yl)diphenylphosphine)ferric chloride was reacted at 90 °C for 18 hours under a carbon monoxide (balloon pressure) atmosphere. After the reaction was complete, the reaction solution was diluted with 60 mL of water, adjusted to pH 5 with saturated citric acid aqueous solution, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and then subjected to column chromatography (petroleum ether:tetrahydrofuran = 3:1) to obtain P-8 (581.51 mg). LCMS (ESI) [M+H-56-18] + =222.1; 1 H NMR (400MHz, DMSO-d6) δ12.71(s,1H),10.01(s,1H),8.15–8.07(m,1H),7.83–7.75(m,2H),6.21(s,1H),1.54(s,6H),1.47(s,9H).

[0179] Preparation Example 3

[0180] Preparation of 3-{[(tert-butoxy)carbonyl]amino}-2-(2-hydroxypropane-2-yl)pyridine-4-carboxylic acid (P-12):

[0181] Step 1: P-9 (5.00 g) was dissolved in dichloromethane (100 mL) solvent, and di-tert-butyl dicarbonate (21.51 g) and N,N-dimethylpyridin-4-amine (6.02 g) were added. The reaction was carried out at 30 °C for 16 hours. After the reaction was completed, the solution was concentrated under vacuum and subjected to column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain P-10 (8.80 g). LCMS (ESI) [M+H]+ =353.2; 1 H NMR (400MHz, DMSO-d6) δ8.64–8.61(m,1H),7.92–7.86(m,1H),7.70–7.64(m,1H),3.82(s,3H),1.31(s,18H).

[0182] Step 2: P-10 (8.80 g) was dissolved in tetrahydrofuran (100 mL), and methylmagnesium bromide (3 M, 41.62 mL) was added dropwise at 0 °C. The reaction was carried out under nitrogen protection at 0 °C for 1 hour. The reaction solution was then quenched with ammonium chloride, extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (petroleum ether:ethyl acetate = 4:1) to obtain P-11 (3.60 g). LCMS (ESI) [M+H] + =253.2; 1 H NMR (400MHz, DMSO-d6) δ9.79(s,1H),8.33–8.28(m,1H),8.15–8.09(m,1H),7.25–7.20(m,1H),6.27(s,1H),1.51(s,6H),1.47(s,9H).

[0183] Step 3: P-11 (2.00 g) was dissolved in tetrahydrofuran (40 mL), and [2-(dimethylamino)ethyl]dimethylamine (2.03 g) was added. Then, n-butyllithium (47.56 mL) was added dropwise at -78 °C. The reaction was carried out under nitrogen protection at -78 °C for 2 hours. Then, solid carbon dioxide (excess dry ice) was added, and the reaction was carried out at -78 °C for 0.5 hours. After the reaction was complete, the mixture was quenched with ammonium chloride aqueous solution, diluted with water (70 mL), adjusted to pH = 5 with saturated citric acid aqueous solution, extracted with ethyl acetate, and purified by reverse-phase column chromatography (C-18 reverse-phase column, 0.01% formic acid-acetonitrile system, 0%-50%) to obtain P-12 (45.00 mg). LCMS (ESI) [M+H] + =297.1; 1 H NMR (400MHz, DMSO-d6) δ9.40(s,1H),8.29(d,J=4.9Hz,1H),7.46(d,J=4.7Hz,1H),6.30(s,1H),1.52(s,6H),1.43(s,9H).

[0184] Preparation Example 4

[0185] Preparation of 5-{[(tert-butoxy)carbonyl]amino}-6-(2-hydroxypropyl-2-yl)pyridine-3-carboxylic acid (P-17):

[0186] Step 1: P-13 (2.00 g) was dissolved in ultra-dry tetrahydrofuran (30 mL). The reaction solution was purged with nitrogen and the reaction temperature was lowered to -78 °C. Methyllithium (27.05 mL) was added dropwise, and the mixture was stirred at -78 °C for 1 hour. The reaction solution was quenched with water, then extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 0%–25%) to obtain P-14 (1.70 g). LCMS (ESI) [M+H] + =230.9.

[0187] Step 2: Di-tert-butyl dicarbonate (2.55 g) was added to an anhydrous ethanol (30 mL) solution of P-14 (1.50 g). The reaction mixture was stirred at 50 °C for 72 hours. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 0%–20%) to obtain P-15 (1.40 g). LCMS (ESI) [M+H] + =330.9; 1 H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 8.58 (d, J = 2.0Hz, 1H), 8.25 (d, J = 2.1Hz, 1H), 6.46 (s, 1H), 1.50 (s, 6H), 1.48 (s, 9H).

[0188] Step 3: Triethylamine (3.97 g) and palladium dichloride (2+)bis((cyclopentadien-1-yl)diphenylphosphine)iron (575.97 mg) were added to a methanol (30 mL) solution of P-15 (1.30 g). The reaction mixture was stirred at 80 °C for 16 hours under carbon monoxide protection. The reaction solution was concentrated under reduced pressure, and the crude product was purified by column chromatography (tetrahydrofuran / petroleum ether = 0%–30%) and reversed-phase silica gel column chromatography (0.1% formic acid / water / acetonitrile = 0%–70%) to obtain P-16 (500.00 mg). LCMS (ESI) [M+H] + =311.1; 1 H NMR (400MHz, DMSO-d6) δ10.11(s,1H),9.02(d,J=1.9Hz,1H),8.74(d,J=2.0Hz,1H),6.62(s,1H),3.99(s,3H),1.64(s,6H),1.59(s,9H).

[0189] Step 4: Lithium hydroxide monohydrate (338.01 mg) was added to a mixed solution of P-16 (500.00 mg) in tetrahydrofuran (8 mL), methanol (4 mL), and water (2 mL). The reaction solution was stirred at 25 °C for 2 hours. The reaction solution was diluted with water and then washed with ethyl acetate. The pH of the aqueous phase was adjusted to 5-6 with citric acid, and then extracted with ethyl acetate (15 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by column chromatography (petroleum ether / tetrahydrofuran = 0%–30%) to obtain P-17 (240.00 mg). LCMS (ESI) [M+H] + =297.0; 1 H NMR (400MHz, DMSO-d6) δ13.32(s,1H),9.96(s,1H),8.88(d,J=1.8Hz,1H),8.61(d,J=1.9Hz,1H),6.47(s,1H),1.53(s,6H),1.48(s,9H).

[0190] Preparation Example 5

[0191] Preparation of (E)-4-hydroxynon-2-enal (P-20):

[0192] Step 1: [1,3-bis(2,4,6-trimethylphenyl)imidazolium-2-yl]dichloro{[2-(propyl-2-yloxy)phenyl]methylene}ruthenium (782.02 mg) was added to ultradry dichloromethane (80 mL) containing P-18 (8 g) and 3,3-dimethoxyprop-1-ene (19.12 g). The reaction mixture was purged with nitrogen and stirred at 25 °C for 20 hours. TLC (petroleum ether / ethyl acetate = 3:1) showed that the starting material was completely consumed and new spots were formed. The mixture was concentrated under reduced pressure, and the crude product was purified by column chromatography (petroleum ether / ethyl acetate (0.1% triethylamine) = 20:1-5:1) to obtain P-19 (5.60 g). 1 H NMR(400MHz, CDCl3) δ5.89(dd,J=15.7,6.0Hz,1H),5.66(dd,J=15.7,3.9Hz,1H),4.80(d,J=4.3H z,1H),4.20–4.08(m,1H),3.33(s,6H),1.57–1.50(m,2H),1.35–1.24(m,6H),0.91–0.85(m,3H).

[0193] Step 2: At 0°C, slowly add 3.29 mL of sulfuric acid and 20 mL of water solution to a tetrahydrofuran (80 mL) solution of P-19 (5.00 g). After the addition is complete, raise the reaction solution to 25°C and stir for 2 hours. Quench the reaction solution with saturated sodium bicarbonate solution, then extract with ethyl acetate. Wash the organic phase with saturated brine (150 mL x 2), dry with anhydrous sodium sulfate, filter, and concentrate to obtain P-20. LCMS(ESI)[M+H] + =157.2; 1 H NMR (400MHz, DMSO-d6) δ9.55(d,J=8.1Hz,1H),7.04(dd,J=15.5,4.3Hz,1H),6.16(ddd,J=15.5,8.1,1.6Hz, 1H),5.17(d,J=5.0Hz,1H),4.31–4.21(m,1H),1.57–1.41(m,2H),1.39–1.19(m,6H),0.86(t,J=6.8Hz,3H).

[0194] Preparation Example 6

[0195] Preparation of 5-{[(tert-butoxy)carbonyl]amino}-6-(2-hydroxypropane-2-yl)pyridine-2-carboxylic acid (P-25):

[0196] Following the preparation method of Example 3 (P-12), P-9 was replaced with P-21 to obtain P-25. LCMS(ESI)[M+H] + =297.2. 1 H NMR (400MHz, DMSO-d6) δ12.75(s,1H),10.16(s,1H),8.51(d,J=8.6Hz,1H),7.93(d,J=8.6Hz,1H),6.45(s,1H),1.55(s,6H),1.48(s,9H).

[0197] Example 1

[0198] Preparation of (2S)-2-({2-[5-amino-6-(2-hydroxypropane-2-yl)pyridin-3-carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}carbamoyl)-3-(3-methanesulfonylphenyl)propionic acid (compound II-1):

[0199] Step 1: Add ditert-butyl dicarbonate (1.26 g) and sodium carbonate (1.63 g) to a 1-1 (950 mg) solution of dichloromethane (20 mL). Stir the mixture at room temperature for 1 hour. After the reaction is complete, add water for treatment. Extract the combined organic phases with dichloromethane. After washing the organic phases, dry them with anhydrous sodium sulfate and concentrate under reduced pressure to obtain 1-2 (1.3 g).

[0200] Step 2: Add (dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylazonium hexafluoro-λ to a solution of 1-2 (1 g) of dimethyl sulfoxide (25 mL). 5 - Phosphazene (1.65 g) was stirred for 10 minutes. Triethylamine (1.17 g) and compound c (1.28 g) were then added to the reaction mixture, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, water (60 mL) was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed, and dried. The mixture was then evaporated to dryness under reduced pressure, and the crude product was separated by silica gel column chromatography (EA / PE = 20%) to obtain 1-3 (1.45 g).

[0201] Step 3: Add trifluoroacetic acid (1 mL) to the 1-3 (1.5 g) dichloromethane (3 mL) solution, stir at room temperature for 1 hour, and evaporate the solvent under reduced pressure to obtain product 1-4 (1.2 g).

[0202] Step 4: Add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylazonium hexafluoro-λ to a solution of P-17 (198.23 mg, 45% purity) in N,N-dimethylformamide (6 mL). 5 - Phosphazene (317.95 mg) was stirred at room temperature for 10 minutes. Then, triethylamine (270.77 mg) and 1-4 (400 mg) were added to the reaction solution, and the reaction was continued at room temperature for 2 hours. After the reaction was completed, water (20 mL) was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed, dried, and evaporated under reduced pressure. The crude product was separated by silica gel column chromatography (EA / PE = 30%) to obtain 1-5 (130 mg).

[0203] Step 5: Add trifluoroacetic acid (1 mL) to a 200 mg solution of dichloromethane (3 mL). Stir the reaction mixture at room temperature for 1 hour. After the reaction is complete, neutralize with saturated sodium bicarbonate solution, extract with dichloromethane, wash and dry the organic phase, and rotary evaporate under reduced pressure to obtain crude product 1-6 (140 mg).

[0204] Step 6: Add potassium hydroxide (2 mL, 1 mol / L) to a 110 mg solution of tetrahydrofuran (2 mL), and stir the reaction mixture at room temperature for 1 hour. Adjust the pH to approximately 5 with 1 M hydrochloric acid, rotary evaporate the organic solvent under reduced pressure, and preparatively separate compound II-1 (61.2 mg, 60%) by LCMS (ESI) [M+H] + =648.9. 1 H NMR (400MHz, DMSO-d6) δ7.83(s,1H),7.73(d,J=7.9Hz,2H),7.64(d,J=7.6Hz,1H),7.52(t,J=7.6Hz,1H),7.43(s,1H),6.99(s,1H), 5.76(s,2H),5.52(s,1H),4.72(s,2H),4.50(s,1H),3.82(s,1H),3.61(s,2H),3.32(s,1H),3.13(s,3H),2.78(s,2H),1.50(s,6H).

[0205] Example 2

[0206] Preparation of (2S)-2-({2-[3-amino-4-(2-hydroxypropane-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-2):

[0207] Step 1: Add P-4 (157.8 mg, 1 eq) to an 8 mL reaction flask, then add DMF (3 mL) and [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylhexafluoro-λ 5 - Phospholipids (1.25 eq), after stirring for 10 minutes, were added to triethylamine (4 eq) and 2-1 (1 eq). The reaction was allowed to proceed at room temperature for 2 hours. After the reaction was completed, the reaction solution was added to water, extracted with ethyl acetate, washed, dried, and the solvent was removed by vacuum evaporation to obtain crude product 2-2 (340 mg).

[0208] Step 2: Add 2-2 (340 mg, 1 eq) to an 8 mL reaction flask, add dichloromethane (3 mL), add trifluoroacetic acid (1 mL) at 0°C, react at 0°C for 1 hour. After the reaction is complete, add the reaction solution to a saturated sodium bicarbonate solution, extract with dichloromethane, wash the organic phase and dry, and evaporate the solvent under reduced pressure to obtain crude product 2-3 (300 mg, crude product).

[0209] Step 3: Add potassium hydroxide (2 mL, 1 M) to a solution of 2-3 (300 mg, crude product) in tetrahydrofuran (2 mL), and stir the reaction mixture at room temperature for 1 hour. Adjust the pH to approximately 5 with 1 M hydrochloric acid, evaporate the organic solvent, and preparatively separate compound II-2 (32 mg) by LCMS (ESI) [M+H]. + =648.28. 1 H NMR (400MHz, DMSO-d6) δ8.88(s,1H),7.85(d,J=1.9Hz,1H),7.76(d,J=7.8Hz,1H ),7.66(d,J=7.7Hz,1H),7.56(t,J=7.7Hz,1H),7.40(s,1H),7.07(d,J=7.9Hz,1 H),6.65(s,1H),6.51(s,1H),5.61(s,2H),5.29(s,1H),4.70(s,3H),3.63(s,2H ),3.15(s,3H),3.04(dd,J=14.1,9.9Hz,1H),2.75(d,J=6.0Hz,2H),1.51(s,6H).

[0210] Example 3

[0211] Preparation of (2S)-2-({2-[4-amino-3-(2-hydroxypropane-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-3):

[0212] Step 1: Add P-8 (148.17 mg, 1 eq) to an 8 mL reaction flask, then add DMF (3 mL), [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylhexafluoro-λ 5 - Phospholipids (1.25 eq), after stirring for 10 minutes, triethylamine (4 eq) and 3-1 (1 eq) were added, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, the reaction solution was added to water, extracted with ethyl acetate, the organic phase was washed and dried, and the solvent was removed by vacuum evaporation to obtain crude product 3-2 (300 mg).

[0213] Step 2: Add 3-2 (300 mg) to an 8 mL reaction flask, add dichloromethane (3 mL), add trifluoroacetic acid (1 mL) at 0°C, react at 0°C for 1 hour. After the reaction is complete, add the reaction solution to a saturated sodium bicarbonate solution, extract with dichloromethane, wash the organic phase and dry, evaporate the solvent under reduced pressure to obtain crude product 3-3 (220 mg).

[0214] Step 3: Add potassium hydroxide (2 mL, 1 M) to a solution of 3-3 (220 mg, crude) in tetrahydrofuran (2 mL), and stir the reaction mixture at room temperature for 1 hour. Adjust the pH to approximately 5 with 1 M hydrochloric acid, evaporate the organic solvent, and preparatively separate compound II-3 (81 mg) by chromatography. LCMS (ESI) [M-18] + =630.28. 1 H NMR(400MHz,DMSO-d6)δ12.90(s,1H),8.93(s,1H),7.86(d,J=1.9Hz,1H),7.80–7.73(m,1H) ,7.67(d,J=7.7Hz,1H),7.56(t,J=7.7Hz,1H),7.36(s,1H),7.15(d,J=2.0Hz,1H),7.09(dd,J =8.2,2.0Hz,1H),6.63(d,J=8.2Hz,1H),5.88(s,2H),5.30(s,1H),4.75(s,1H),4.67(s,2H), 3.71(t,J=6.0Hz,2H),3.15(s,3H),3.03(dd,J=14.1,10.2Hz,1H),2.75(s,2H),1.50(s,6H).

[0215] Example 4

[0216] Preparation of (2S)-2-({2-[3-amino-2-(2-hydroxypropyl-2-yl)pyridin-4-carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}carbamoyl)-3-(3-methanesulfonylphenyl)propionic acid (compound II-4):

[0217] Step 1: Add P-12 (61.7 mg, 0.83 eq) to an 8 mL reaction flask, add DMF (3 mL), stir for 10 minutes, then add triethylamine (4 eq) and 4-1 (1 eq), react at room temperature for 2 hours. After the reaction is complete, add the reaction solution to water, extract with ethyl acetate, wash the organic phase and dry, and evaporate the solvent under reduced pressure to obtain crude product 4-2 (150 mg).

[0218] Step 2: Add 4-2 (150 mg, crude product) to an 8 mL reaction flask, add dichloromethane (3 mL), add trifluoroacetic acid (1 mL) at 0 degrees Celsius, react at 0 degrees Celsius for 1 hour, after the reaction is completed, add the reaction solution to a saturated sodium bicarbonate solution, extract with dichloromethane, wash the organic phase and dry, evaporate the solvent under reduced pressure to obtain crude product 4-3 (100 mg).

[0219] Step 3: Add potassium hydroxide (2 mL, 1 M) to a 100 mg solution of tetrahydrofuran (2 mL), and stir the reaction mixture at room temperature for 1 hour. Adjust the pH to approximately 5 with 1 M hydrochloric acid, evaporate the organic solvent, and preparatively separate compound II-4 (35 mg) by chromatography. LCMS (ESI) [M+1] + =649.27. 1 H NMR(400MHz, DMSO-d6)δ8.29(s,1H),7.82(t,J=1.8Hz,1H),7.79–7.69(m,2 H),7.64(dt,J=7.7,1.4Hz,1H),7.52(t,J=7.7Hz,1H),7.45(s,1H),6.92(s, 1H),5.62(s,2H),4.78(s,1H),4.47(s,1H),3.87(s,1H),3.49(s,2H),3.26 (d,J=4.9Hz,1H),3.13(s,3H),3.14–3.06(m,1H),2.76(s,2H),1.51(s,6H).

[0220] Example 5

[0221] Preparation of (2S)-2-({2-[6-amino-5-(2-hydroxypropane-2-yl)pyridin-3-carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}carbamoyl)-3-(3-methanesulfonylphenyl)propionic acid (compound II-10):

[0222] Step 1: Add [chloro(dimethylamino)methylene]dimethylhexafluoro-λ to a 4 mL acetonitrile solution of compound 6b (92.22 mg). 5 - Phospholipids (197.81 mg) and 1-methyl-1H-imidazole (154.36 mg). Then 5-1 (220 mg) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, evaporated under reduced pressure, and the crude product was separated by silica gel column chromatography to obtain 5-2 (220 mg).

[0223] Step 2: Add methanol (4 mL) and water (2 mL) to 5-2 (220 mg). Add lithium hydroxide (40.72 mg), and stir the reaction solution at 50°C for 1 hour. Filter the reaction solution and prepare II-10 (120 mg) by preparative chromatographic separation. LCMS (ESI) [M+1] + =649.33. 1H NMR (400MHz, DMSO-d6) δ8.65(d,J=7.0Hz,1H),8.00(d,J=2.1Hz,1H),7.84(d,J=1.9Hz,1H),7. 75(dt,J=7.9,1.4Hz,1H),7.69–7.62(m,1H),7.54(t,J=7.7Hz,1H),7.43–7.35(m,2H),6.56(s, 2H),5.51(s,1H),4.70(s,2H),4.64(td,J=8.4,4.8Hz,1H),3.73(t,J=6.1Hz,2H),3.29(dd,J=1 4.0, 4.7Hz, 1H), 3.14 (s, 3H), 3.06 (dd, J=14.0, 9.1Hz, 1H), 2.77 (t, J=6.0Hz, 2H), 1.49 (s, 6H).

[0224] Example 6

[0225] Preparation of ((2S)-2-({2-[5-amino-6-(2-hydroxypropane-2-yl)pyridin-2-carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}carbamoyl)-3-(3-methanesulfonylphenyl)propionic acid (compound II-11):

[0226] Referring to Example 4, intermediate P-12 was replaced with P-25 to obtain compound II-11. LCMS(ESI)[M+1] + =649.33. 1 H NMR (400MHz, DMSO-d6) δ8.57(s,1H),7.84(s,1H),7.75(d,J=7.8Hz,1H),7.65(d,J=7. 7Hz,1H),7.54(t,J=7.7Hz,1H),7.42(d,J=8.3Hz,2H),7.02(d,J=8.2Hz,1H),6.06(s,2 H),5.58(s,1H),5.08(s,1H),4.73(s,1H),4.59(s,1H),3.99(s,1H),3.84(s,1H),3.27 (d,J=4.8Hz,1H),3.14(s,3H),3.07(dd,J=13.9,8.8Hz,1H),2.83(s,2H),1.51(s,6H).

[0227] Example 7

[0228] Preparation of (2S)-3-[4-amino-3-(2-hydroxypropyl-2-yl)phenyl]-2-{[2-(1-benzofuran-6-carbonyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl]formamido}propionic acid (compound II-19):

[0229] Step 1: Add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylhexafluoro-λ to a solution of 2-[(tert-butoxy)carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (548.9 mg) in N,N-dimethylformamide (10 mL). 5 - Phospholipids (753.5 mg) were stirred for 0.166 hours. Triethylamine (481.3 mg) and 7-1 (400 mg) were then added to the reaction mixture, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, water was added, followed by extraction with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and evaporated under reduced pressure. The crude product was separated by silica gel column chromatography to obtain 7-2 (500 mg).

[0230] Step 2: Add 1 mL of trifluoroacetic acid to the 7-2 (500 mg) dichloromethane solution, react at room temperature for 1 hour, adjust the pH to 8 with sodium bicarbonate aqueous solution, and extract with ethyl acetate to obtain 7-3 (350 mg).

[0231] Step 3: Add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylammonium hexafluoro-λ to a solution of 1-benzofuran-6-carboxylic acid (178.89 mg) in N,N-dimethylformamide (5 mL). 5 - Phospholipids (524.38 mg). The reaction mixture was stirred for 0.166 hours. Triethylamine (334.93 mg) and 7-3 (530 mg) were then added to the reaction mixture. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, water was added, followed by extraction with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and evaporated under reduced pressure. The crude product was separated by silica gel column chromatography to obtain 7-4 (340 mg).

[0232] Step 4: Add methanol (2 mL) and water (1 mL) to 7-4 (300 mg, 1 eq.). Add lithium hydroxide (57.53 mg), and stir the reaction solution at 50°C for 1 hour. Filter the reaction solution to obtain II-19 (135 mg). LCMS (ESI) [M+H] + =609.9. 1H NMR(400MHz, DMSO-d6)δ8.70(s,1H),8.12(d,J=2.2Hz,1H),7.78–7.69(m,2H),7.33( d,J=8.0Hz,2H),7.05(d,J=2.2Hz,1H),6.91(d,J=2.0Hz,1H),6.84(dd,J=8.1,2.0Hz, 1H),6.48(d,J=8.0Hz,1H),5.18(s,2H),4.75(s,2H),4.47(q,J=7.5Hz,1H),3.67(s, 3H), 2.91 (dd, J=14.0, 5.4Hz, 1H), 2.80 (dd, J=8.6, 5.1Hz, 3H), 1.47 (d, J=1.9Hz, 6H).

[0233] Example 8

[0234] Preparation of (2S)-2-({2-[5-amino-4-(2-hydroxypropane-2-yl)pyridin-2-carbonyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}carbamoyl)-3-(3-methanesulfonylphenyl)propionic acid (compound II-20):

[0235] Step 1: Add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylhexafluoro-λ to a solution of 5-{[(tert-butoxy)carbonyl]amino}-4-(2-hydroxypropane-2-yl)pyridine-2-carboxylic acid (207.46 mg) 5 - Phospholipids (332.76 mg). The reaction mixture was stirred for 0.166 h. Triethylamine (334.93 mg) and 8-1 (330 mg) were then added to the reaction mixture. The mixture was stirred at room temperature for 2 h. After the reaction was complete, water was added, followed by extraction with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and evaporated under reduced pressure. The crude product was separated by silica gel column chromatography to obtain 8-2 (450 mg).

[0236] Step 2: Add trifluoroacetic acid (1 mL) to a solution of 8-2 (450 mg) in dichloromethane (4 mL), react at room temperature for 1 hour, adjust the pH to 8 with sodium bicarbonate aqueous solution, extract with ethyl acetate to obtain 8-3 (380 mg).

[0237] Step 3: Add methanol (2 mL) and water (1 mL) to 8-3 (400 mg). Add lithium hydroxide (57.53 mg), and stir the reaction solution at 50°C for 1 hour. Filter the reaction solution to obtain II-20 (258 mg). LCMS (ESI) [M+1] + =649.35.1 H NMR (400MHz, DMSO-d6) δ8.42(s,1H),7.92(s,1H),7.83(d,J=1.9Hz,1H),7.73(dt,J=7.8,1 .5Hz,1H),7.65(d,J=7.7Hz,1H),7.53(t,J=7.7Hz,1H),7.42(s,1H),7.35(s,1H),5.99(s,2 H),5.50(s,1H),4.92(s,1H),4.73(s,1H),4.54(q,J=7.2,6.6Hz,1H),3.86(s,2H),3.28(d d,J=13.9,4.8Hz,1H),3.13(s,3H),3.08(dd,J=13.8,8.2Hz,1H),2.78(s,2H),1.48(s,6H).

[0238] Example 9

[0239] Preparation of (2S)-2-({2-[4-amino-2-chloro-5-(2-hydroxypropane-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-21):

[0240] Referring to Example 4, intermediate P-12 was replaced with 4-{[(tert-butoxy)carbonyl]amino}-2-chloro-5-(2-hydroxypropane-2-yl)benzoic acid to obtain compound II-21. LCMS (ESI) [M-18] + =644.29. 1 H NMR (400MHz, DMSO-d6) δ8.84(t,J=6.1Hz,1H),8.62(s,1H),7.83(t,J=1.8Hz,1H),7.73(dt,J= 7.8,1.5Hz,1H),7.65(dt,J=7.7,1.5Hz,1H),7.53(t,J=7.7Hz,1H),7.30(s,2H),7.12–7.04(m ,2H),6.96(dd,J=8.1,1.9Hz,1H),5.59(s,2H),5.31(s,1H),4.61–4.54(m,1H),4.38(d,J=5.9 Hz,2H),3.28(dd,J=13.9,4.7Hz,1H),3.13(s,3H),3.05(dd,J=13.9,8.8Hz,1H),1.50(s,6H).

[0241] Example 10

[0242] Preparation of (2S)-2-({2-[4-amino-2-fluoro-5-(2-hydroxypropane-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-24):

[0243] Referring to Example 4, intermediate P-12 was replaced with 4-{[(tert-butoxy)carbonyl]amino}-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoic acid to obtain compound II-24 (117 mg). LCMS (ESI) [M-18] + =648.3. 1 H NMR (400MHz, DMSO-d6) δ8.70 (s, 1H), 7.84 (d, J = 2.0Hz, 1H), 7.75 (d, J = 7.9Hz, 1H), 7.66(d,J=7.7Hz,1H),7.54(t,J=7.7Hz,1H),7.39(s,1H),7.00(d,J=7.9Hz,1H),6. 42(d,J=12.5Hz,1H),6.06(s,2H),5.34(s,1H),4.65(s,3H),3.81(s,1H),3.55(s, 1H), 3.14 (s, 3H), 3.05 (dd, J=14.0, 9.2Hz, 1H), 2.71 (t, J=5.6Hz, 2H), 1.47 (s, 6H).

[0244] Example 11

[0245] Preparation of ((2S)-2-({2-[2-amino-3-(2-hydroxypropane-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-37)

[0246] Referring to Example 5, 6-amino-5-(2-hydroxypropane-2-yl)pyridine-3-carboxylic acid was replaced with compound 1e to obtain compound II-37 (49 mg).

[0247] LCMS(ESI)[M+H] + =647.9. 1H NMR (400MHz, DMSO-d6) δ8.37(s,1H),7.82(s,1H),7.73(d,J=7.7Hz,1H),7.64(d,J=7.7Hz, 1H),7.52(t,J=7.7Hz,1H),7.35(s,1H),7.12(d,J=7.7Hz,1H),6.92(d,J=7.5Hz,1H),6.57 (t,J=7.6Hz,1H),5.57(s,2H),5.41(s,1H),4.68(s,2H),4.51(s,1H),3.65(s,2H),3.28(d d,J=13.8,4.8Hz,1H),3.13(s,3H),3.11–3.04(m,1H),2.75(d,J=6.6Hz,2H),1.52(s,6H).

[0248] Example 12

[0249] Preparation of (S)-2-(2-(4-amino-5-(2-hydroxypropyl-2-yl)-2-(trifluoromethyl)benzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (compound II-26)

[0250] Referring to Example 4, intermediate P-12 was replaced with 4-((tert-butoxycarbonyl)amino)-5-(2-hydroxypropyl-2-yl)-2-(trifluoromethyl)benzoic acid to obtain compound II-26 (157.46 mg).

[0251] LCMS(ESI)[M+H] + =716.3. 1 H NMR (400MHz, DMSO-d6) δ8.36 (s, 1H), 7.82 (d, J = 2.1

[0252] Hz,1H),7.73(d,J=7.7Hz,1H),7.64(d,J=7.7Hz,1H),7.52(t,J=7.7Hz,1H),7.32(d,J=94.8Hz,1H),7.06–6.89(m,2H),6 .05(s,2H),5.47(d,J=8.9Hz,1H),4.89–4.06(m,4H),3.26(d,J=4.8Hz,3H),3.12(s,4H),2.85–2.56(m,2H),1.48(s,6H).

[0253] Example 13

[0254] Preparation of (2S)-2-({2-[4-amino-5-(2-hydroxypropyl-2-yl)-2-methoxybenzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compound II-29)

[0255] Step 1: Under nitrogen protection and an ice bath, methyl magnesium bromide (5.96 g) was added to 40 mL of tetrahydrofuran containing 15-1 (2.6 g). After the addition was complete, the mixture was slowly brought to room temperature and stirred for 2 hours. After the reaction was completed, the reaction solution was quenched with ammonium chloride solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under low pressure to obtain the target compound 15-2 (2.6 g).

[0256] Step 2: Add di-tert-butyl dicarbonate (4.36 g) to 50 mL of ethanol solution of 15-2 (2.6 g), heat to 55 °C and stir overnight. After the reaction is complete, evaporate the reaction solution under low pressure and separate the crude product by silica gel column chromatography to obtain 15-3 (3.4 g).

[0257] Step 3: Compound 15-3 (1g), 1,1,3-trioxo-2,3-dihydro-1λ 6 2-Benzothiazole-2-carboxaldehyde (2.34 g), sodium carbonate (735.55 mg), 4-(diphenylphosphono)butyl[diphenylphosphine] (355.15 mg), and palladium acetate (124.64 mg) were added to the solvent N,N-dimethylformamide (15 mL), followed by triethylsilane (645.57 mg). The mixture was heated to 80 °C and stirred overnight. After the reaction was complete, the solvent was removed under reduced pressure, and the crude product was separated by silica gel column chromatography to obtain 15-4 (500 mg).

[0258] Step 4: Add 2 mL of an aqueous solution of sodium dihydrogen phosphate (581.75 mg) and sodium chlorite (219.26 mg) to 8 mL of a tert-butanol solution of 15-4 (300 mg). After the addition is complete, stir the mixture at room temperature overnight. After the reaction is complete, concentrate the reaction solution and separate the crude product by silica gel column chromatography to obtain 15-5 (50 mg).

[0259] Step 5: Add 1 mL of trifluoroacetic acid to 5 mL of a 15-5 (300 mg) dichloromethane solution. After stirring for half an hour, the reaction is complete. The reaction solution is then directly evaporated to dryness to obtain 15-6 (200 mg).

[0260] Step 6: Add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylammonium hexafluoro-λ to 15-6 (90 mg) N,N-dimethylformamide (4 mL).5 - Phosphoryl hydrazine (204.31 mg) and triethylamine (108.75 mg) were stirred at room temperature for 15 minutes, and then (2S)-2-[(5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl)formamido]-3-(3-methanesulfonylphenyl)propionate benzyl ester (201.14 mg) was added. The reaction was stirred for another hour. After the reaction was completed, the reaction solution was quenched with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure. The crude product was separated by silica gel column chromatography to obtain 15-7 (150 mg).

[0261] Step 7: Add 0.5 mL of an aqueous solution of sodium hydroxide (37.75 mg) to 2 mL of an ethanol solution of 15-7 (150 mg), heat to 80 °C and stir for 5 hours. After the reaction is complete, adjust the pH to about 9 with an aqueous solution of citric acid under ice bath, filter, and prepare II-29 (4.6 mg) from the filtrate.

[0262] LCMS(ESI)[M+1] + =677.9. 1 H NMR (400MHz, DMSO-d6) δ8.81(s,1H),7.86(s,1H),7.76(d,J=7.7Hz,1H),7.67(d,J=7.5Hz,1H),7.56(t,J=7.7Hz,1H),7.41(s,1H),6.84(s,1H),6. 31(s,1H),5.82(s,2H),5.10(s,1H),4.70(s,2H),4.41(s,1H),3.69(s,2H ),3.55(s,2H),3.15(s,3H),3.09–3.00(m,2H),2.73(s,3H),1.45(s,6H).

[0263] Example 14

[0264] Preparation of (S)-2-(2-(4-amino-5-(2-hydroxypropyl-2-yl)-2-methylbenzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (compound II-27)

[0265] Step 1: 4-((tert-Butoxycarbonyl)amino)-5-(2-hydroxypropyl-2-yl)-2-methylbenzoic acid (80.0 mg) was dissolved in dichloromethane (3.0 mL), then 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (120 mg), N,N-diisopropylethylamine (40 mg), and 16-1 (160 mg) were added, and the reaction was continued at 25 °C for 4 hours. LCMS was used to determine if the reaction was complete. Water (30.0 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, and the reaction solution was evaporated to dryness to obtain the crude compound. The target product 16-2 (110 mg) was obtained by column chromatography using a petroleum ether-ethyl acetate system (petroleum ether:ethyl acetate = 1:1). LCMS (ESI) [M+H] + =852.24

[0266] Step 2: Dissolve 16-2 (110 mg) in dichloromethane (5.0 mL), add it to a solution dissolved in dichloromethane (10.0 mL), then cool to 0 °C, slowly add trifluoroacetic acid (35 mg), and continue the reaction for 2.0 hours. LCMS analysis showed complete reaction of the starting material and product formation. The reaction solution was directly evaporated to dryness to obtain the crude target product 16-3 (110 mg). LCMS (ESI) [M+H] + =777.94

[0267] Step 3: Dissolve 16-3 (110 mg) in anhydrous ethanol (4.0 mL), add water (4.0 mL), then add potassium hydroxide (30 mg), heat to 70 °C, and continue the reaction for 16.0 hours. LCMS analysis showed that the reaction proceeded completely and products were formed. After filtration and purification, the target compound II-27 (12.3 mg) was obtained. LCMS (ESI) [M+H] + =662.34; 1 H NMR (400MHz, DMSO-d6) δ7.82 (s, 1H), 7.79 (s, 1H), 7.70 (d, J = 8.0Hz, 1H), 7.6 2(d,J=8.0Hz,1H),7.49(t,J=7.8Hz,1H),7.43–7.15(m,1H),6.78(s,1H),6. 46(s,1H),5.58(s,2H),5.25(s,1H),4.82–4.47(m,2H),4.34–4.30(m,1H),3 .29–3.24(m,2H),3.17–3.12(m,5H),2.69(s,2H),2.02(s,3H),1.46(s,6H).

[0268] Example 15

[0269] Preparation of (S)-2-(2-(4-amino-3-(2-hydroxypropyl-2-yl)benzoyl)-5-chloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (compound II-39)

[0270] Step 1: Dissolve 17-1 (330 mg) in N,N-dimethylformamide (5 mL), add benzyl 2-amino-3-(3-methanesulfonylphenyl)propionate hydrochloride (391.5 mg), stir at room temperature for 10 minutes, add N,N-diisopropylethylamine (0.41 mL) and [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylene]dimethylammonium hexafluoro-λ 5 - Phosphate (804.95 mg), reacted at room temperature for 1 hour. After the reaction was complete, the reaction solution was diluted with water (30 mL), extracted with ethyl acetate (30 mL x 3), the organic phases were combined, washed with water (30 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated, and then subjected to column chromatography (petroleum ether:tetrahydrofuran = 3:1) to give the target compound 17-2 (630 mg). LCMS (ESI) [M+H-56] + =571.1;

[0271] Step 2: Dissolve 17-2 (650 mg) in dichloromethane (4 mL), add trifluoroacetic acid (1 mL), and react at room temperature for 2 hours. After the reaction is complete, evaporate the solvent from the reaction mixture, and purify by prep-HPLC (C-18, 0.1% NH4HCO3 in H2O / ACN) to obtain the target compound 17-3 (500 mg). LCMS (ESI) [M+H] + =527.1.

[0272] Step 3: At room temperature, 17-3 (400 mg) was dissolved in N,N-dimethylformamide (8 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, (577.17 mg)), 4-{[(tert-butoxy)carbonyl]amino}-3-(2-hydroxypropyl-2-yl)benzoic acid (224.15 mg) and N,N-diisopropylethylamine (DIEA, 0.38 mL) were added. The mixture was stirred at room temperature for 1 hour. The reaction solution was quenched with water (20 mL), extracted three times with ethyl acetate (20 mL), and the combined organic phases were washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by rapid chromatography (Silica gel, tetrahydrofuran:petroleum ether = 2:1) to obtain the target compound 17-4 (350 mg). LCMS (ESI) [M+H] +=804.2.

[0273] Step 4: At room temperature, 17-4 (350 mg) was dissolved in dichloromethane (6 mL), and trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 0.5 hours. The reaction solution was concentrated and purified by rapid chromatography (Silica gel, 100% EA) to obtain the target compound 17-5 (150 mg). LCMS (ESI) [M+H-18] + =686.3.

[0274] Step 5: At room temperature, 17-5 (150 mg) was dissolved in tetrahydrofuran (2.5 mL), and sodium hydroxide aqueous solution (1 M, 2.5 mL) was added. The mixture was stirred at room temperature for 3 hours. The reaction solution was adjusted to pH 7 with hydrochloric acid aqueous solution (1 M) and then purified by Prep-HPLC (C18, 10 mmol / L FA in water, MeCN) to obtain the target compound II-39 (71.77 mg). LCMS (ESI) [M+H] + =614.3;

[0275] 1 H NMR(400MHz,DMSO-d6)δ8.45(br.s,1H),7.85(s,1H),7.77(d,J=7.7Hz,1H),7.6 5(d,J=7.6Hz,1H),7.57(t,J=7.7Hz,1H),7.20–7.14(m,2H),7.13–7.05(m,2H), 6.63(d,J=8.2Hz,1H),5.87(Brs,2H),5.31(s,1H),4.68(s,2H),4.63–4.53(m,1 H),3.73(t,J=5.6Hz,2H),3.21–3.00(m,5H),2.81(t,J=5.9Hz,2H),1.50(s,6H).

[0276] Example 16

[0277] Preparation of (2S)-2-(2-(4-amino-3-(2-hydroxypropyl-2-yl)benzoyl)-5-chloro-4-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (compounds II-35, II-40):

[0278] Step 1: Dissolve 18-4 (3.6 g) in tetrahydrofuran (50 mL), add borane tetrahydrofuran (56.34 mL, 1 M) at 0 °C, and react at 70 °C for 2 hours. After the reaction is complete, quench the reaction solution with methanol (40 mL), concentrate, and obtain the target compound 18-5 (2.3 g) by column chromatography (dichloromethane:methanol = 10:1). LCMS (ESI) [M+H] + =228.1.

[0279] Step 2: Dissolve 18-5 (2.3 g) in dichloromethane (30 mL), add triethylamine (2.27 mL) and trifluoroacetic anhydride (2.25 mL) at 0 °C, and react at room temperature for 2 hours. After the reaction is complete, concentrate the reaction solution, dilute with water (50 mL), extract with ethyl acetate (50 mL x 3), combine the organic phases, dry with anhydrous sodium sulfate, filter and concentrate, and purify by column chromatography (petroleum ether: tetrahydrofuran = 10:1) to obtain the target compound 18-6 (2.6 g). LCMS (ESI) [M+H] + =324.1; 1 H NMR (400MHz, CDCl3) δ7.65–7.57(m,1H),7.42–7.39(m,1H),7.35(t,J=7.7Hz, 1H), 6.24 (s, 1H), 3.78–3.73 (m, 1H), 3.67–3.52 (m, 2H), 1.32 (d, J = 6.9Hz, 3H).

[0280] Step 3: At 0°C, 18-6 (3g) was added to acetic acid (9mL) and sulfuric acid (6mL), and stirred at room temperature for 2 hours. Paraformaldehyde (1.39g) was then added, and the reaction was carried out at 50°C for 16 hours. After the reaction was complete, the reaction solution was added to water (100mL) under ice bath conditions, extracted with ethyl acetate (80mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and then subjected to column chromatography (petroleum ether:tetrahydrofuran = 10:1) to obtain the target compound 18-7 (500mg). LCMS (ESI) [M+H] + =336.0.

[0281] Step 4: Dissolve 18-7 (650 mg) in methanol (4 mL) and sodium hydroxide aqueous solution (4 mL), and react at room temperature for 16 hours. Concentrate the reaction system directly to dryness to obtain crude product (18-8) for the next step.

[0282] LCMS(ESI)[M+H] + =226.1.

[0283] Step 5: Dissolve 18-8 (650 mg, crude product) in dichloromethane (10 mL), add triethylamine (437.19 mg) and di-tert-butyl dicarbonate (942.94 mg) at room temperature, and react at room temperature for 4 hours. After the reaction, adjust the pH to 5-6 with dilute hydrochloric acid (1 M), extract three times with ethyl acetate (10 mL), combine the organic phases, backwash with saturated sodium chloride aqueous solution, dry with anhydrous sodium sulfate, filter, concentrate to dryness, and purify the crude product using rapid chromatography (silica, pure tetrahydrofuran) to obtain the target product 18-9 (220 mg). LCMS (ESI) [M+H-56] + =270.2. 1 H NMR (400MHz, DMSO-d6) δ13.35(s,1H),7.53(d,J=8.0Hz,1H),7.24(d,J=8.1Hz,1H),4.93(d,J=17.3Hz,1H),4.4 0–4.18(m,1H),3.26(s,1H),3.14(d,J=14.2Hz,1H),2.98(d,J=12.7Hz,1H),1.44(s,9H),1.13(d,J=5.0Hz,3H).

[0284] Step 6: Dissolve 18-9 (200 mg) in N,N-dimethylformamide (3 mL), add benzyl(S)-2-amino-3-(3-(methylsulfonyl)phenyl)propionate hydrochloride (249.76 mg), stir for 10 minutes at room temperature, add N,N-diisopropylethylamine (0.3 mL) and [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-yloxy})methylimene]dimethylammonium hexafluoro-λ 5 - Phosphoramide (466.84 mg) was reacted at room temperature for 1 hour. After the reaction was complete, the reaction solution was diluted with water (10 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with water (10 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated, and then subjected to column chromatography (petroleum ether:tetrahydrofuran = 1:1) to give the target compound 18-10 (400 mg). LCMS (ESI) [M+H-56] + =585.3; 1HNMR (400MHz, CDCl3) δ7.83–7.75(m,2H),7.44–7.30(m,8H),7.05(d,J=7.9Hz,1H),6.58(t,J=7.7Hz,1H),5.25–5.15(m,3H ),4.29–4.18(m,1H),3.77–3.72(m,3H),3.49–3.42(m,1H),3.34–3.21(m,2H),2.97(s,3H),1.50(s,9H),1.23–1.15(m,3H).

[0285] Step 7: At room temperature, 18-10 (410 mg) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL), and stirred for 1 hour at room temperature. Product formation was monitored by LCMS. After the reaction, the pH was adjusted to 7 with sodium bicarbonate (1 M), and the mixture was extracted three times with ethyl acetate (10 mL). The combined organic phases were washed with saturated sodium chloride aqueous solution (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product 18-11 (410 mg). LCMS (ESI) [M+H] + =541.4.

[0286] Step 8: To a solution of 18-11 (300 mg) of N,N-dimethylformamide (5 mL), add 4-{[(tert-butoxy)carbonyl]amino}-3-(2-hydroxypropyl-2-yl)benzoic acid (163.75 mg), N,N-diisopropylethylamine (0.3 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (316.24 mg). React at room temperature for 1 hour. After the reaction is complete, add water (20 mL), extract with ethyl acetate (20 mL x 3), combine the organic phases, wash with saturated sodium chloride solution (20 mL x 3), dry the organic phase, concentrate, and purify by rapid chromatography (petroleum ether:tetrahydrofuran = 1:1) to obtain the target compound 18-12 (350 mg). LCMS (ESI) [M+H] + =818.1

[0287] Step 9: Add 2 mL of dioxane chloride solution (4 M) to a solution of 18-12 (350 mg) of dioxane (2 mL), and allow the mixture to react at room temperature for 1 hour. After the reaction is complete, concentrate the reaction solution and purify it by rapid chromatography (petroleum ether: tetrahydrofuran = 1:5) to obtain the target compound 18-13 (150 mg). LCMS (ESI) [M+H] + =718.1

[0288] Step 10: Add 1 mL of sodium hydroxide aqueous solution (1M) to a 130 mg (18-13) tetrahydrofuran solution and react at room temperature for 1 hour. After the reaction is complete, adjust the pH to 7 with 1M hydrochloric acid, extract with ethyl acetate (20 mL * 3), combine the organic phases, wash with 20 mL of saturated sodium chloride solution, dry the organic phase, concentrate, and separate and purify by prep-HPLC (C18, 10 mmol / L formic acid in water, MeCN) to obtain the target compounds II-35 and II-40 (35.59 mg). LCMS (ESI) [M + H] + =628.3;1H NMR (400MHz, DMSO-d6) δ8.08(s,1H),7.81(s,1H),7.75(d,J=7.4Hz,1H),7.62(d, J=7.5Hz,1H),7.54(t,J=7.5Hz,1H),7.19–7.05(m,4H),6.64(d,J=8.2Hz,1H),5.8 3(s,2H),5.31(s,1H),5.14–4.82(m,1H),4.52–4.36(m,2H),4.36–3.95(m,1H),3 .24–3.19(m,2H),3.18–3.04(m,5H),1.50(d,J=3.1Hz,6H),1.11(d,J=6.9Hz,3H).

[0289] Example 17

[0290] Preparation of (2S)-2-({2-[4-amino-3-(2-hydroxypropyl-2-yl)benzoyl]-5,7-dichloro-4-methyl-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (compounds II-33, II-36):

[0291] Step 1: Dissolve 19-1 (5 g) in tetrahydrofuran (50 mL), add triethylamine (8.62 g) and 3-bromo-1-propene (3.44 g), and stir the mixture at 50 °C for 16 hours. After the reaction is complete, filter the reaction solution, concentrate the filtrate, and perform column chromatography (PE / THF = 10 / 1) on the crude product to obtain the target compound 19-2 (2.2 g). LCMS (ESI) [M+H] + =216.3. 1HNMR(400MHz,DMSO-d6)δ7.43(t,J=1.9Hz,1H),7.38(d,J=1.9Hz,2H),5.95–5.7 6(m,1H),5.22–5.02(m,2H),3.67(s,2H),3.16–3.04(m,2H),2.48–2.42(m,1H).

[0292] Step 2: Dissolve 19-2 (2.2 g) in trifluoromethanesulfonic acid (20 mL) and stir at 70 °C for 16 hours. After the reaction is complete, cool to room temperature, pour the reaction solution into ice water, adjust the pH of the solution to 7-8 with saturated sodium carbonate solution, extract with ethyl acetate (30 mL * 3), combine the organic phases, backwash with saturated sodium chloride aqueous solution, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and perform column chromatography (PE / THF = 3 / 1) on the crude product to obtain the target compound 19-3 (1.5 g). LCMS (ESI) [M+H] + =216.4. 1 H NMR (400MHz, DMSO-d6) δ7.36(s,1H),7.13(s,1H),3.95–3.74(m,2H),2.93–2.79(m,3H),1.23(d,J=6.9Hz,3H).

[0293] Step 3: Dissolve 19-3 (1.4 g) in tetrahydrofuran (20 mL), add triethylamine (1.97 g) and di-tert-butyl dicarbonate (2.12 g) at room temperature, and stir the mixture at room temperature for 16 hours. After the reaction is complete, concentrate the reaction solution, and perform column chromatography (PE / THF = 10 / 1) on the crude product to obtain the target compound 19-4 (1.6 g). LCMS (ESI) [M+H-56+41] + =301.0.

[0294] Step 4: Dissolve 19-4 (1.6 g) in tetrahydrofuran (20 mL), add 2.5 M n-butyllithium (3.04 mL) at -78 °C, and stir for 1 hour at this temperature. Add a small piece of dry ice and stir at room temperature for 15 hours. After the reaction is complete, add saturated ammonium chloride (10 mL), extract with ethyl acetate (25 mL * 2), combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate the filtrate, and perform column chromatography (C18, 0.03% TFA in H2O / ACN) to obtain the target compound 19-5 (300 mg). LCMS (ESI) [M + H - 56 + 41] + =345.3.

[0295] Step 5: Dissolve 19-5 (200 mg) and benzyl(S)-2-amino-3-(3-(methylsulfonyl)phenyl)propionate hydrochloride (308.02 mg) in N,N-dimethylformamide (5 mL), and add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-oxy})methylimene]dimethylammonium hexafluoro-λ at room temperature. 5 Phosphoramide (422.21 mg) and diisopropylethylamine (215.26 mg) were stirred at room temperature for 16 hours. After the reaction was complete, water (15 mL) was added to the reaction solution, the mixture was filtered, and the filter cake was dried to obtain the crude product. The crude product was subjected to column chromatography (PE / THF = 2 / 1) to obtain the target compound 19-6 (200 mg). LCMS (ESI) [M+H-56] + =619.0.

[0296] Step 6: Dissolve 19-6 (150 mg) in dichloromethane (5 mL), add trifluoroacetic acid (1.5 mL) at room temperature, and stir for 2 hours at room temperature. After the reaction is complete, concentrate the reaction solution to obtain the target compound 19-7 (crude product, 120 mg). LCMS (ESI) [M+H] + =575.2.

[0297] Step 7: Dissolve 19-7 (120 mg) and 4-{[(tert-butoxy)carbonyl]amino}-3-(2-hydroxypropyl-2-yl)benzoic acid (73.9 mg) in N,N-dimethylformamide (3 mL), and add [(dimethylamino)({3H-[1,2,3]triazolo[4,5-b]pyridin-3-oxy})methylidene]dimethylammonium hexafluoro-λ at room temperature. 5 Phosphoramide (158.57 mg) and N,N-diisopropylethylamine (80.85 mg) were stirred at room temperature for 3 hours. After the reaction was complete, water (10 mL) was added to the reaction solution, the mixture was filtered, and the filter cake was dried to obtain the crude product. The crude product was subjected to column chromatography (PE / THF = 2 / 1) to obtain the target compound 19-8 (200 mg). LCMS (ESI) [M+H] + =852.1.

[0298] Step 8: Dissolve 19-8 (100 mg) in 1,4-dioxane (2 mL), add 4M dioxane hydrochloride (2 mL), and stir at room temperature for 2 hours. After the reaction is complete, concentrate the reaction solution to obtain the crude product. The crude product is then subjected to column chromatography (PE / EA = 1 / 2) to obtain the target compound 19-9 (80 mg). LCMS (ESI) [M+H-18] + =734.1.

[0299] Step 9: Dissolve 19-9 (80 mg) in tetrahydrofuran (1 mL), add 1 N NaOH (1 mL) solution at room temperature, and stir the mixture at room temperature for 1 hour. LCMS showed the reaction was complete. Adjust the pH of the mixture to 6 with 1 N HCl solution, then filter. The filtrate was subjected to pre-HPLC (0.1% FA in H2O / ACN) to obtain the target compounds II-33 and II-36 (63.44 mg). LCMS (ESI) [M+H-18] + =644.1. 1 H NMR (400MHz, DMSO-d6) δ13.09–12.42(m,1H),9.06–8.91(m,1H),7.86(s,1H),7.77(d,J=7.5Hz,1H),7.67(d ,J=7.8Hz,1H),7.57(t,J=7.7Hz,1H),7.35(s,1H),7.12(d,J=1.8Hz,1H),7.09–7.02(m,1H),6.63(d,J=8.2H z,1H),5.83(s,2H),5.30(s,1H),5.07–4.91(m,1H),4.81–4.70(m,1H),4.49–4.36(m,1H),4.28–4.02(m,1H ),3.29–3.17(m,3H),3.15(d,J=1.5Hz,3H),3.07–2.97(m,1H),1.50(d,J=2.7Hz,6H),1.09(d,J=6.5Hz,3H).

[0300] Example 18

[0301] Preparation of (S)-2-(2-(4-amino-2-cyclopropyl-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (compound II-32):

[0302] Step 1: 4-Amino-2-cyclopropyl-5-(2-hydroxypropyl-2-yl)benzoic acid (100.0 mg) was dissolved in dichloromethane (5.0 mL), followed by the addition of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (193.93 mg), N,N-diisopropylethylamine (164.79 mg), and 20-1 (253.94 mg). The reaction was carried out at 25 °C for 2 hours. After the reaction was complete, water (20.0 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (20.0 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed under reduced pressure. The target compound 20-2 (85 mg) was obtained by silica gel column chromatography (petroleum ether:ethyl acetate = 1:4). LCMS (ESI) [M+H] + =778.31

[0303] Step 2: Dissolve 20-2 (65 mg) in anhydrous ethanol (2.0 mL), add water (2.0 mL), then add lithium hydroxide monohydrate (2.4 mg), and react at 25 °C for 2.0 h. After the reaction is complete, filter the reaction solution and purify by reverse preparative chromatography to obtain the target compound II-32 (38.0 mg). LCMS (ESI) [M+H] + =688.34; 1 H NMR(400MHz, DMSO-d6)δ7.78(s,1H),7.69(d,J=12.0Hz,1H),7.64–7.54(m,2H),7.47(t,J =7.7Hz,1H),7.22(s,1H),6.76(s,1H),6.13(s,1H),5.53(s,2H),5.20(s,1H),4.77(d,J= 40.3Hz,1H),4.45(s,1H),4.19–4.15(m,1H),3.50(s,1H),3.29–3.16(m,2H),3.11(s,3H) ,2.69(s,2H),1.72(d,J=23.5Hz,1H),1.44(s,6H),1.24(s,1H),0.83(s,2H),0.54(s,2H).

[0304] Example 19

[0305] Preparation of (S)-2-(2-(4-amino-3-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (compound II-43):

[0306] Referring to Example 4, intermediate P-12 was replaced with 21-1 to obtain compound II-43 (17.66 mg). LCMS (ESI) [M+H] + =666.3; 1 H NMR (400MHz, DMSO-d6) δ8.41 (s, 1H), 7.83 (d, J = 2.0Hz, 1H), 7.76–7.71 (m, 1H), 7.65(d,J=7.7Hz,1H),7.53(t,J=7.7Hz,1H),7.37(s,1H),7.09(dd,J=11.3,1.8 Hz,1H),7.01(s,1H),5.77(s,2H),5.51(s,1H),4.68(s,2H),4.52(s,1H),3.71 (s,2H),3.27(d,J=7.3Hz,2H),3.13(s,4H),2.77(d,J=5.8Hz,2H),1.51(s,6H).

[0307] Example 20

[0308] Preparation of (S)-2-(2-(4-amino-2-fluoro-3-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-45):

[0309] Referring to Example 16, 4-((tert-Butoxycarbonyl)amino)-5-(2-hydroxypropyl-2-yl)-2-methylbenzoic acid was replaced with 23-1 to obtain the target compound II-45 (22.84 mg). LCMS (ESI) [M+1] + =665.8. 1 H NMR (400MHz, DMSO-d6) δ8.54(s,1H),7.83(t,J=1.8Hz,1H),7.74(dt,J=7.7,1.5Hz,1H),7.65(dt,J=7.8,1.4Hz,1H),7.53(t,J=7.7Hz ,1H),6.89(s,2H),6.44(d,J=8.3Hz,1H),6.27(s,2H),5.66(s,2H),4.80–4.35(m,4H),3.13(s,5H),2.70(s,3H),1.55(d,J=3.5Hz,6H)

[0310] Example 21

[0311] Preparation of (2S)-2-({2-[4-amino-2,3-difluoro-5-(2-hydroxypropyl-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (II-47):

[0312] Referring to Example 20, 4-amino-2-cyclopropyl-5-(2-hydroxypropyl-2-yl)benzoic acid was replaced with 25-1 to obtain compound II-47 (106.16 mg). LCMS (ESI) [M+H-18] + =666.4. 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),8.84(s,1H),7.85(s,1H),7.76(d,J=7.8 Hz,1H),7.66(d,J=7.9Hz,1H),7.56(t,J=7.7Hz,1H),7.48–7.28(m,1H),6.93–6 .80(m,1H),6.00(s,2H),5.54(s,1H),4.83–4.51(m,3H),3.90–3.50(m,2H),3.2 9–3.26(m,1H),3.14(s,3H),3.08–3.00(m,1H),2.77–2.66(m,2H),1.48(s,6H).

[0313] Example 22

[0314] Preparation of (S)-2-(2-(4-amino-2,5-difluoro-3-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (II-48):

[0315] Step 1: 26-1 (4.5 g) was dissolved in N,N-dimethylformamide (50 mL), and 1-bromopyrrolidine-2,5-dione (4.71 g) was added at 0 °C. The reaction was allowed to proceed for 1 hour at room temperature. LCMS showed that after the reaction was complete, saturated sodium thiosulfate aqueous solution (100 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (100 mL * 3), the organic phases were combined, washed with saturated brine (100 mL * 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The filtrate was then purified by rapid chromatography (Silica gel, petroleum ether:ethyl acetate = 5:1) to obtain compound 26-2 (6.2 g). LCMS (ESI) [M+H] + =268.0; 1HNMR(400MHz,DMSO-d6)δ7.70–7.61(m,1H),6.62(s,2H),3.85(s,3H).

[0316] Step 2: 26-2 (3g) was dissolved in tetrahydrofuran (50mL), and 3M methylmagnesium bromide (22.55mL) was added dropwise under nitrogen protection at 0℃. The reaction was then carried out at room temperature for 2 hours. After the reaction was complete as shown by TLC, saturated ammonium chloride solution (40mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (100mL*3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 26-3 (2.7g) was then purified by rapid chromatography (Silica gel, PE:EA = 5:1). LCMS (ESI) [M+H-18] + =248.0; 1 HNMR (400MHz, DMSO-d6) δ7.37–7.28 (m, 1H), 6.00 (s, 2H), 5.92 (s, 1H), 1.56 (d, J = 3.8Hz, 6H).

[0317] Step 3: 26-3 (500 mg) was dissolved in dimethyl sulfoxide (10 mL) and water (5 mL). 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (275.75 mg) and triethylamine (2.6 mL) were added. The reaction was carried out at 90 °C for 16 hours under a carbon monoxide atmosphere. LCMS analysis confirmed the reaction was complete. The reaction solution was cooled, and water (20 mL) was added. Impurities were back-extracted with ethyl acetate (20 mL). The aqueous phase was adjusted to pH 4 with dilute hydrochloric acid and extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by rapid chromatography (Silica gel, PE:EA = 1:3) to obtain compound 26-4 (150 mg). LCMS (ESI) [M+H-18] + =214.1; 1 HNMR (400MHz, DMSO-d6) δ12.49(s,1H),7.38–7.28(m,1H),6.60(s,2H),5.92(s,1H),1.57(d,J=3.5Hz,6H).

[0318] Step 4: At 0°C, (2S)-2-[(5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl)formamido]-3-(3-methanesulfonylphenyl)propionate benzyl ester hydrochloride (200 mg), triethylamine (101.54 mg), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (190.77 mg) were added to a solution of 26-4 (77.33 mg) in N,N-dimethylformamide (5 mL), and stirred at room temperature for 1 hour. The reaction solution was quenched with water (15 mL) and extracted with ethyl acetate (15 mL * 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by rapid chromatography (Silica gel, petroleum ether:ethyl acetate = 4:1) to obtain compound 26-5 (200 mg). LCMS(ESI)[M+H] + =774.0.

[0319] Step 5: At room temperature, 26-5 (180 mg) was dissolved in tetrahydrofuran (2.5 mL), and sodium hydroxide aqueous solution (1 M, 2.5 mL) was added. The mixture was stirred at room temperature for 1 hour. The pH of the reaction solution was adjusted to 7 with hydrochloric acid aqueous solution (1 M), and water (10 mL) was added. The mixture was extracted with ethyl acetate (10 mL * 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by Prep-HPLC (10 mmol / L FA in water, MeCN) to obtain compound II-48 (131.44 mg). LCMS (ESI) [M + H] + =684.1; 1 H NMR (400MHz, DMSO-d6) δ12.84(s,1H),8.96(s,1H),7.86(s,1H),7.77(d,J=7.4Hz,1H), 7.67(d,J=7.8Hz,1H),7.56(t,J=7.8Hz,1H),7.49–7.25(m,1H),7.05–6.88(m,1H),6.18 (s,2H),5.88(s,1H),4.81–4.69(m,2H),4.60–4.40(m,1H),3.93–3.73(m,1H),3.58–3. 46(m,1H),3.28(s,2H),3.14(s,3H),3.08–2.99(m,1H),2.74–2.70(m,1H),1.56(s,6H).

[0320] Example 23

[0321] Preparation of (2S)-2-({2-[4-amino-2,6-difluoro-3-(2-hydroxypropyl-2-yl)benzoyl]-5,7-dichloro-1,2,3,4-tetrahydroisoquinoline-6-yl}formamido)-3-(3-methanesulfonylphenyl)propionic acid (II-49):

[0322] Referring to Example 26, replacing 26-1 with compound 27-1 yielded compound II-49 (160.15 mg). LCMS (ESI) [MH] - =682.2. 1 H NMR(400MHz,DMSO-d6)δ12.87(s,1H),9.09–8.91(m,1H),7.86(s,1H),7.77(d,J=7.8Hz,1H),7.67 (d,J=7.4Hz,1H),7.57(t,J=7.7Hz,1H),7.47–7.26(m,1H),6.50(s,2H),6.37–6.24(m,1H),5.71( d,J=17.0Hz,1H),4.83–4.71(m,2H),4.48(s,1H),3.99–3.76(m,1H),3.57–3.51(m,1H),3.29–3.2 4(m,1H),3.14(s,3H),3.07–2.96(m,1H),2.78–2.70(m,1H),2.68–2.64(m,1H),1.59–1.48(m,6H).

[0323] Example 24

[0324] Preparation of (S)-2-(2-(4-amino-3-(2-hydroxypropyl-2-yl)benzoyl)-8-chloro-2,3-dihydro-1H-pyrrolo[3,2,1-ij]quinazolin-7-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-51):

[0325] Step 1: 29-11 (110 mg) was added to N,N-dimethylformamide (4 mL), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (125 mg) and N,N-diisopropylethylamine (133 mg) were added under ice bath conditions. The mixture was stirred under ice bath conditions for 5 minutes. 29-12 (132 mg) was added under ice bath conditions, and the mixture was stirred at room temperature for 1 hour. Product formation was detected by LCMS. The reaction mixture was added to water (20 mL), extracted with ethyl acetate (20 mL x 2), and the organic phase was dried over anhydrous sodium sulfate. The crude product was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain the target compound 29-13 (200 mg). LCMS (ESI) [M+H] + =652.3. 1 H NMR(400MHz, DMSO-d6)δ8.92(d,J=8.0Hz,1H),7.90(t,J=1.8Hz,1H),7.79(s ,1H),7.68(s,1H),7.58(t,J=7.7Hz,1H),7.51–7.25(m,7H),7.00(s,1H),5. 94(d,J=3.0Hz,1H),5.65(d,J=2.7Hz,2H),5.21(s,2H),4.96(ddd,J=10.7,8 .0,4.9Hz,1H),4.81(s,2H),3.37(d,J=4.9Hz,1H),3.10(s,4H),1.38(s,9H).

[0326] Step 2: 200 mg of 29-13 was added to 2 mL of dichloromethane, followed by 0.4 mL of trifluoroacetic acid. The mixture was stirred at room temperature for 3 hours. LCMS analysis confirmed complete consumption of the starting material. The reaction solution was then added to 20 mL of saturated sodium carbonate aqueous solution and extracted with a dichloromethane:methanol ratio of 10:1 (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the target compound 29-14 (155 mg, crude product). LCMS (ESI) [M+H] + =552.2.

[0327] Step 3: Add 29-15 (75 mg) to N,N-dimethylformamide (2 mL), then add 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (114 mg), stir at room temperature for 10 minutes, then add a solution of 29-14 (155 mg) and N,N-diisopropylethylamine (121 mg) in N,N-dimethylformamide (2 mL), and stir at room temperature for 1 hour. LCMS analysis confirmed complete consumption of the starting materials. The reaction mixture was added to water (20 mL), extracted with ethyl acetate (20 mL x 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 29-16 (240 mg, crude). LCMS (ESI) [M+H-56-18] + =773.3.

[0328] Step 4: Add 200 mg of 29-16 to 2 mL of dichloromethane, add 0.5 mL of trifluoroacetic acid under ice bath conditions, and stir for 4 hours under ice bath conditions. LCMS confirmed complete consumption of the starting material. Add the reaction solution to 20 mL of saturated sodium carbonate aqueous solution, extract with dichloromethane:methanol = 10:1 (20 mL x 2), and concentrate under reduced pressure to obtain compound 29-17 (170 mg, crude product). LCMS (ESI) [M+H-18] + =773.3.

[0329] Step 5: Add 29-17 (170 mg) to methanol (2 mL) and water (1 mL), then add lithium hydroxide monohydrate (50 mg) and stir at room temperature for 1 hour. LCMS confirmed complete consumption of the starting material. The reaction solution was purified by Prep-HPLC (C18, 10 mmol / L NH4HCO3 in water, MeCN) to obtain the target compound II-51 (12.58 mg). LCMS (ESI) [M+H-18] + =639.3; 1 H NMR (400MHz, DMSO-d6) δ7.80–7.69(m,2H),7.61(d,J=7.6Hz,1H),7.49(p,J=7.5Hz,2H),6.99(t,J=13.8Hz,2H),6.45(d,J=12.7Hz,1H),6.19(d,J =14.6Hz,2H),5.74(s,2H),5.34(s,1H),4.88(s,2H),4.34(d,J=6.0Hz,1 H),3.39(s,1H),3.18(dd,J=13.5,6.5Hz,2H),3.05(s,3H),1.43(s,6H).

[0330] Example 25

[0331] Preparation of (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (II-67):

[0332] Step 1: At 0℃, acetyl chloride (13.13 g) was slowly added dropwise to a solution of 2-(3-methoxyphenyl)ethyl-1-amine (30-1, 23 g) and triethylamine (23.09 g) in dichloromethane (230 mL), and the reaction was allowed to proceed for 1 hour. The reaction was detected as complete by LCMS. Water (230 mL) was added to the reaction solution to separate the organic phase. The solution was washed with 0.5 M hydrochloric acid aqueous solution (230 mL) and saturated brine (230 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (23.5 g). LCMS (ESI) [M+H] + =194.1.

[0333] Step 2: At 0°C, phosphorus oxychloride (9.52 mL) was added to a solution of N-(3-methoxyphenylethyl)acetamide (30-2, 10 g) in acetonitrile (100 mL), and the reaction was carried out at 75°C for 3 hours. The reaction was confirmed by LCMS. After cooling to room temperature, the reaction solution was concentrated, and the residue was added to a 2M potassium phosphate solution (200 mL). The mixture was stirred for 30 minutes and extracted with ethyl acetate (100 mL * 3). The organic phases were combined, washed with saturated brine (160 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (8.8 g). LCMS (ESI) [M + H] + =176.2; 1 H NMR(400MHz, CDCl3) δ7.42(d,J=8.5Hz,1H),6.79(dd,J=8.5,2.6Hz,1H),6.70(d,J=2.5Hz ,1H),3.84(s,3H),3.63(td,J=7.4,1.4Hz,2H),2.72–2.65(m,2H),2.35(t,J=1.4Hz,3H).

[0334] Step 3: At 0℃, sodium borohydride (4.75 g) was added in batches to a methanol (200 mL) solution of 6-methoxy-1-methyl-3,4-dihydroisoquinoline (30-3, 11 g). The mixture was heated to 25℃ and reacted for 16 hours. The reaction was confirmed by LCMS. The pH of the reaction solution was adjusted to 3 with 1M hydrochloric acid, the methanol was removed by concentration, the residue was adjusted to pH 8 with 1M sodium hydroxide solution, and extracted with ethyl acetate (60 mL * 3). The organic phases were combined, washed with saturated brine (80 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (9.5 g). LCMS (ESI) [M+H] + =178.3; 1 H NMR (400MHz, CDCl3) δ7.05(d,J=8.6Hz,1H),6.76(dd,J=8.6,2.6Hz,1H),6.63(d,J=2.5Hz,1H),4.27(q,J=6.6Hz,1H),3.78(s,3H ),3.42–3.33(m,1H),3.20–3.11(m,1H),3.03(ddd,J=13.7,8.3,5.6Hz,1H),2.87(dt,J=16.7,4.9Hz,1H),1.59(d,J=6.7Hz,3H).

[0335] Step 4: 150 g of 47% hydrobromic acid aqueous solution was added to 6-methoxy-1-methyl-1,2,3,4-tetrahydroisoquinoline (30-4, 15.35 g), and the reaction was carried out at 100 °C for 16 hours. The reaction was confirmed by LCMS. The reaction solution was concentrated, and the residue was slurried in 100 mL of ethyl acetate, filtered, and the solid was collected to obtain the target compound (18.3 g). LCMS (ESI) [M+H] + =164.2; 1 HNMR(400MHz,DMSO-d6)δ9.46(s,1H),9.26(s,1H),8.87(s,1H),7.09(d,J=8.5Hz,1H),6.68(dd,J=8.5,2.4Hz,1H) ,6.59(d,J=2.3Hz,1H),4.45(dd,J=10.0,6.4Hz,1H),3.44–3.24(m,2H),3.00–2.85(m,2H),1.54(d,J=6.8Hz,3H).

[0336] Step 5: Potassium carbonate (73597.26 mg) was added to 1-methyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrobromide (30-5, 6.5 g) in dichloromethane (60 mL) and methanol (6 mL), and reacted at 25 °C for 2 hours. The mixture was filtered, the filtrate was collected, concentrated, and the residue was diluted with ethyl acetate (60 mL). Hydrochloric acid gas in ethyl acetate solution (60 mL, 4 M) was added, and the mixture was reacted at 25 °C for 1 hour. The mixture was filtered, and the solid was collected to obtain the target compound (4.61 g). LCMS (ESI) [M+H] + =164.1; 1 H NMR (400MHz, DMSO-d6) δ9.86(s,1H),9.58(s,1H),9.31(s,1H),7.08(d,J=8.5Hz,1H),6.69(dd,J=8.5,2.3Hz,1H) ,6.59(d,J=2.1Hz,1H),4.39(d,J=4.6Hz,1H),3.33(s,1H),3.21(s,1H),3.04–2.81(m,2H),1.55(d,J=6.7Hz,3H).

[0337] Step 6: p-Toluenesulfonic acid hydrate (43.92 mg) was added to acetonitrile (60 mL) containing 1-methyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrochloride (30-6, 4.61 g) and 1-chloro-2,5-pyrrolidone (9.25 g). The reaction was carried out at 25 °C for 3 hours. The reaction was confirmed by LCMS. The reaction solution was filtered, and the filter cake was washed with acetonitrile (50 mL). The filter cake was collected to obtain the target compound (4.29 g). LCMS (ESI) [M+H] + =232.0.

[0338] Step 7: Triethylamine (4.85 g) was added to 5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrochloride (30-7, 4.29 g), di-tert-butyl dicarbonate (4.18 g), and dichloromethane (50 mL). The reaction was carried out at 25 °C for 16 hours. The reaction was confirmed by LCMS. Water (100 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (50 mL x 3). The organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by rapid chromatography (Silicagel, PE / THF = 10 / 1) to obtain the target compound (3.26 g). LCMS (ESI) [M+H-56+ACN] + =317.0.

[0339] Step 8: At -78℃ under a nitrogen atmosphere, trifluoromethanesulfonic anhydride (7.3 g) was slowly added dropwise to a solution of 5,7-dichloro-6-hydroxy-1-methyl-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (30-8, 4.3 g) and 2,6-dimethylpyridine (4.16 g) in dichloromethane (50 mL). After the addition was complete, the temperature was raised to 25℃ and the reaction was carried out for 16 hours. The reaction was detected by LCMS. The reaction was quenched with water (100 mL), extracted with dichloromethane (50 mL * 3), the organic phases were combined, washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was purified by rapid chromatography (Silca gel, PE / THF = 10 / 1) to obtain the target compound (3.24 g). LCMS (ESI) [M + H - 56 + ACN] + =449.0.

[0340] Step 9: Palladium acetate (115.09 mg) was added to 5,7-dichloro-1-methyl-6-(((trifluoromethyl)sulfonyl)oxy)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester (30-9, 2.38 g), bis(2-(diphenylphosphono)cyclopentan-2,4-dien-1-yl)iron (284.19 mg), and triethylamine (2.59 g) in methanol (20 mL). The reaction was carried out at 70 °C for 16 hours under a carbon monoxide atmosphere. The reaction was detected by LCMS to indicate completion. After cooling to room temperature, the reaction solution was filtered, the filtrate was concentrated, and the residue was purified by rapid chromatography (Silica gel, PE / THF = 20 / 1) to obtain the target compound (340 mg). LCMS (ESI) [M+H-56+ACN] + =359.1; 1 H NMR(400MHz,DMSO-d6)δ7.61(s,1H),5.17(s,1H),4.09(s,1H),3.91(s,3H), 3.19(s,1H),2.82(d,J=16.7Hz,1H),2.68–2.59(m,1H),1.45–1.38(m,12H).

[0341] Step 10: At 25℃, trifluoroacetic acid (2 mL) was added to a solution of 2-(tert-butyl)-6-methyl-5,7-dichloro-1-methyl-3,4-dihydroisoquinoline-2,6(1H)-dicarboxylate (30-10, 150 mg) in dichloromethane (8 mL). The reaction was allowed to proceed for 2.0 h at this temperature. LCMS analysis confirmed the reaction was complete and successful. The reaction solution was diluted with water (50 mL), and the pH was adjusted to 7 with saturated sodium bicarbonate solution. Extraction was then performed with dichloromethane (40 mL * 3). The organic phases were combined, washed with saturated sodium chloride solution (30 mL * 3), dried, and concentrated to obtain the target compound (80 mg). LCMS (ESI) [M+H] + =274.0.

[0342] Step 11: At 25℃, 4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoic acid (62.22 mg), methylimidazolium (47.62 mg), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (163.76 mg) were added to a solution of methyl 5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid methyl ester (30-11 mg, 47.62 mg) in acetonitrile (15 mL). The reaction was carried out at this temperature for 2.0 h. The reaction was confirmed to be complete by LCMS. The reaction solution was diluted with water (50 mL), extracted with dichloromethane (30 mL x 3), the organic phases were combined, washed with saturated sodium chloride solution (20 mL x 3), dried, concentrated, and purified by rapid chromatography (petroleum ether:tetrahydrofuran = 10:1) to obtain the target compound (65 mg). LCMS(ESI)[M+H-18] + =451.3.

[0343] Step 12: At 25°C, lithium hydroxide (25.52 mg) was added to a methanol (3 mL), water (12 mL) solution of methyl 2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid ester (30-12, 100 mg). The reaction was carried out at 100°C for 5 hours. LCMS analysis confirmed the reaction was complete and successful. The reaction solution was diluted with water (30 mL), and the pH was adjusted to 6 with 1.0 M hydrochloric acid. Extraction was performed with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated sodium chloride solution (20 mL x 3), dried, and concentrated to obtain the target compound (65 mg). LCMS (ESI) [M+H] + =455.1.

[0344] Step 13: At 25°C, add N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (133.62 mg) and triethylamine (36.43 mg) to a solution of 2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (30-13,80 mg), (2S)-2-amino-3-(3-methanesulfonylphenyl)propionate benzyl ester hydrochloride (66.6 mg) in N,N-dimethylformamide (10 mL). React at 50°C for 5 hours. LCMS analysis indicates the reaction is complete and successful. The reaction mixture was diluted with water (50 mL), extracted with dichloromethane (20 mL x 3), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (20 mL x 3), dried, concentrated, and purified by rapid chromatography (petroleum ether:tetrahydrofuran = 10:2) to obtain the target compound (55 mg). LCMS (ESI) [M+H-18] + =752.3.

[0345] Step 14: At 25°C, add sodium hydroxide (7.2 mg) to a solution of (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionate (30-14, 50 mg) in water (2 mL) and tetrahydrofuran (8 mL). React at 25°C for 1 hour. LCMS analysis indicates the reaction is complete and successful. The reaction solution was diluted with water (20 mL), and the pH was adjusted to 6 with hydrochloric acid aqueous solution (1.0 M). Extraction was performed with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated sodium chloride solution (20 mL x 3), dried, and concentrated to obtain the crude product. The crude product was purified by Prep-HPLC (C18, 10 mmol / L FA in water, MeCN) to obtain the target compound II-67 (25 mg). LCMS (ESI) [M + H-18] + =662.2; 1H NMR (400MHz, DMSO-d6) δ12.90(br.s,1H),8.94(s,1H),7.86(s,1H),7.77(d,J=7.7Hz,1H),7. 67(d,J=7.2Hz,1H),7.58-7.54(m,1H),7.49-7.31(m,1H),6.97(s,1H),6.42(d,J=12.1Hz,1H) ,6.03(s,2H),5.58(s,1H),5.34(s,1H),4.77-4.57(m,2H),3.65-3.32(m,1H),3.28-3.24(m, 1H),3.14(s,3H),3.05-2.97(m,1H),2.79-2.76(m,1H),2.67-2.51(m,1H),1.48-1.42(m,9H).

[0346] Example 26, Example 27

[0347] Preparation of (S)-2-((R)-2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (II-68) and (S)-2-((S)-2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (II-69):

[0348] (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methylsulfonyl)phenyl)propionic acid (II-67) (19 mg, 27.9 mmol) was resolved to give: II-67-P1(II-68 or II-69)LCMS(ESI)[M+H-18] + =662.2; Chiral separation peak time: 1.473 minutes; 1H NMR (400MHz, DMSO-d6) δ8.41(s,1H),7.83(s,1H),7.73(d,J=7.9Hz,1H),7.64(d,J=7.9Hz,1H),7.54(d ,J=7.7Hz,1H),7.43(d,J=50.6Hz,1H),7.02-6.90(m,1H),6.52-6.34(m,1H),6.03(s,2H),5.74-5.44( m,1H),5.39–5.26(m,1H),4.93–4.39(m,2H),3.77–3.48(m,1H),3.25(dd,J=10.6,4.5Hz,1H),3.13(s, 3H), 3.10–3.04(m,1H),2.83–2.73(m,1H),2.61(dd,J=11.9,7.2Hz,1H),1.45(dd,J=16.0,7.7Hz,9H).

[0349] II-67-P2 (II-68 or II-69) (6.0 mg). LCMS(ESI)[M+H-18] + =662.2; Chiral separation peak time: 2.6 minutes; 1 H NMR (400MHz, DMSO-d6) δ7.95(s,1H),7.80(s,1H),7.71(d,J=7.8Hz,1H),7.63(d,J=7.7H z,1H),7.49(t,J=7.7Hz,2H),6.98(d,J=7.0Hz,1H),6.43(d,J=11.7Hz,1H),6.03(s,2H) ,5.45(d,J=86.6Hz,2H),4.86-4.24(m,2H),3.68-3.42(m,1H),3.28-3.25(m,1H),3.19- 3.14(m,1H),3.12(s,3H),2.79(d,J=16.8Hz,1H),2.68-2.57(m,1H),1.50-1.40(m,9H).

[0350] Separation method: Waters SFC 150; Column name: Column specifications: 250*30mm 10μm; Mobile phase A: supercritical carbon dioxide; Mobile phase B: ethanol (+0.1% 7.0M ammonia methanol); Mobile phase ratio A:B = 65:35; Detection wavelength: 214nm; Flow rate: 120ml / min; Column temperature: room temperature; Back pressure: 100bar; Injection volume: 3mL; Cycle time: 5.6min; Solvents: ethanol (preparative grade), supercritical carbon dioxide (food grade); Sample preparation: The sample was dissolved in 12ml of methanol.

[0351] Example 28

[0352] Preparation of (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-70):

[0353] Step 1: At 0℃, acetyl chloride (13.46 g) was slowly added dropwise to a solution of 2-(3-methoxyphenyl)ethyl-1-amine (33-1, 20 g) and triethylamine (20.08 g) in dichloromethane (200 mL), and the reaction was allowed to proceed for 1 hour. The reaction was detected as complete by LCMS. Water (200 mL) was added to the reaction solution to separate the organic phase. The solution was washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (27 g). LCMS (ESI) [M+H] + =208.2.

[0354] Step 2: At 0°C, phosphorus oxychloride (23.08 g) was added to a solution of N-(2-(3-methoxyphenyl)ethyl)propionamide (33-2, 26 g) in acetonitrile (260 mL), and the reaction was carried out at 75°C for 3 hours. The reaction was confirmed by LCMS. After cooling to room temperature, the reaction solution was concentrated, and the residue was added to a sodium bicarbonate solution (500 mL) to adjust the pH to 8. Extraction was performed with ethyl acetate (500 mL * 3), and the organic phases were combined. The mixture was washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (19 g). LCMS (ESI) [M+H] + =190.2.

[0355] Step 3: At 0℃, sodium borohydride (4g) was added in batches to a methanol (180mL) solution of 1-ethyl-6-methoxy-3,4-dihydroisoquinoline (33-3, 10g), and the mixture was heated to 25℃ and reacted for 16 hours. The reaction was confirmed by LCMS. The pH of the reaction solution was adjusted to 3 with 1M hydrochloric acid, the methanol was removed by concentration, the residue was adjusted to pH 8 with 1M sodium hydroxide solution, and extracted with ethyl acetate (60mL*3). The organic phases were combined, washed with saturated brine (80mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the target compound (10g). LCMS (ESI) [M+H] + =192.2.

[0356] Step 4: 118 mL of 48% hydrobromic acid aqueous solution was added to 1-ethyl-6-methoxy-1,2,3,4-tetrahydroisoquinoline (33-4, 10 g), and the reaction was carried out at 100 °C for 16 hours. The reaction was confirmed by LCMS. The reaction solution was concentrated, and the residue was slurried with 100 mL of ethyl acetate, filtered, and the solid was collected to obtain the target compound (11.8 g). LCMS (ESI) [M+H] + =178.2.

[0357] Step 5: Potassium carbonate (117.78 g) was added to dichloromethane (1200 mL) and methanol (80 mL) containing 1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrobromide (33-5, 11 g). The mixture was reacted at 25 °C for 16 hours. The mixture was filtered, the filtrate was collected, concentrated, and the residue was diluted with ethyl acetate (110 mL). An ethyl acetate hydrochloride solution (106.53 mL) was added, and the mixture was reacted at 25 °C for 16 hours. The mixture was filtered, and the solid was collected to obtain the target compound (9.1 g). 1 H NMR (400MHz, DMSO-d6) δ9.79(s,1H),9.58(s,1H),9.19(s,1H),7.06(d,J=8.8Hz,1H),6.68(d,J=8.4Hz,1H) ,6.60(s,1H),4.25(s,1H),3.37-3.18(m,2H),3.07-2.83(m,2H),2.00-1.88(m,2H),1.01(t,J=7.2Hz,3H).

[0358] Step 6: p-Toluenesulfonic acid hydrate (80.11 mg) was added to acetonitrile (120 mL) containing 1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrochloride (33-6.9 g) and 1-chloro-2,5-pyrrolidone (16.87 g). The reaction was carried out at 25°C for 3 hours. The reaction was confirmed by LCMS. The reaction mixture was filtered, and the filter cake was washed with acetonitrile (50 mL). The filter cake was collected to obtain the target compound (10 g). LCMS (ESI) [M+H] + =245.9.

[0359] Step 7: 5,7-Dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-ol hydrochloride (33-7, 10 g) was dissolved in dichloromethane (100 mL), and di-tert-butyl dicarbonate (6.18 g) and triethylamine (10.74 g) were added at room temperature. The reaction was allowed to proceed for 16 hours at room temperature. LCMS analysis confirmed the reaction was complete. The reaction solution was concentrated, and the crude product was subjected to column chromatography (petroleum ether / tetrahydrofuran = 10 / 1) to obtain the target compound (5.2 g). LCMS (ESI) [MH]- = 344.0.

[0360] Step 8: Dissolve 33-8.5 g of 5,7-dichloro-1-ethyl-6-hydroxy-3,4-dihydroisoquinoline-2(1H)-carboxylic acid tert-butyl ester in dichloromethane (50 mL), add 4641.88 mg of 2,6-dimethylpyridine at room temperature, cool to -78 °C under nitrogen atmosphere, add trifluoromethanesulfonic anhydride (8148.45 mg) dropwise, then heat to room temperature and stir for 16 hours. The reaction was confirmed by LCMS. The reaction solution was concentrated, and the crude product was subjected to column chromatography (petroleum ether / tetrahydrofuran = 10 / 1) to obtain the target compound (5.6 g). LCMS (ESI) [M+H-56+41] + =462.8.

[0361] Step 9: Dissolve tert-butyl 5,7-dichloro-1-ethyl-6-((trifluoromethyl)sulfonyl)oxy)-3,4-dihydroisoquinoline-2(1H)-carboxylic acid (33-9, 2 g) in methanol (40 mL), add bis(2-(diphenylphosphono)cyclopentan-2,4-dien-1-yl)iron (231.73 mg), palladium acetate (93.84 mg), and triethylamine (2114.87 mg, 20.9 mmol, 5 eq), and stir at 70 °C for 16 hours under carbon monoxide (balloon pressure). The reaction was confirmed by LCMS. The reaction solution was filtered, the filtrate was concentrated, and the crude product was subjected to column chromatography (PE / THF = 10 / 1) to obtain the target compound (560 mg). LCMS (ESI) [M+H-56+41] + =372.9.

[0362] Step 10: Dissolve 2-(tert-butyl)6-methyl-5,7-dichloro-1-ethyl-3,4-dihydroisoquinoline-2,6(1H)-dicarboxylate (33-10, 560 mg) in dichloromethane (24 mL) and trifluoroacetic acid (6 mL), and stir at room temperature for 2 hours. The reaction was confirmed by LCMS. The reaction solution was concentrated, mixed with diethyl ether (30 mL), and filtered to obtain the product (410 mg). LCMS (ESI) [M+H] + =288.0.

[0363] Step 11: Methyl 5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (33-11, 510 mg) was dissolved in acetonitrile (5 mL), and N-methylimidazolium (581.2 mg) was added. The mixture was stirred at room temperature for 20 minutes. N,N,N',N'-tetramethylchloromethanemidazone hexafluorophosphate (744.85 mg) and 4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoic acid (490.54 mg) were added, and the mixture was stirred at room temperature for 2 hours. The reaction was monitored by LCMS and found to be complete. The reaction solution was diluted with water (30 mL), extracted with ethyl acetate (30 mL x 3), washed with sodium chloride aqueous solution (30 mL), dried over anhydrous sodium sulfate, filtered, and evaporated to dryness to obtain the crude target compound (460 mg). LCMS (ESI) [M+H] + =483.0

[0364] Step 12: Lithium iodide (731.06 mg) was added to pyridine (10 mL) containing methyl 2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (33-12, 440 mg). The reaction was carried out at 125°C for 2 hours, and the reaction was confirmed by LCMS. After cooling to room temperature, the target compound (365 mg) was obtained by reversing the reaction in a C18 system (0.1% FA in water / ACN = 40% / 60%). LCMS (ESI) [M+H] + =469.0.

[0365] Step 13: Add 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (437.48 mg) to 2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (33-13, 360 mg), 2-amino-3-(3-methanesulfonylphenyl)propionate benzyl ester hydrochloride (425.55 mg), ethyl bis(2-propyl)amine (297.4 mg) in N,N-dimethylformamide (10 mL), react at 25°C for 1 hour, and the reaction is completed by LCMS. The reaction mixture was added to water (20 mL), extracted with ethyl acetate (15 mL * 3), the organic phases were combined, washed with saturated brine (15 mL * 3), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was passed through a reversed-phase system (C18, 0.1% FA in water / ACN = 40% / 60%) to give the target compound (475 mg). LCMS (ESI) [M + H-18] + =766.0.

[0366] Step 14: Add 6 mL of 1 M sodium hydroxide aqueous solution to 6 mL of tetrahydrofuran containing benzyl (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionate (33-14, 450 mg). React at 25°C for 3 hours. The reaction was confirmed by LCMS. Adjust the pH to 7 with 1 M hydrochloric acid, concentrate the reaction solution, and extract the residue by prep-HPLC (C18, 0.1% FA in water / MeCN) to obtain the target compound II-70 (205 mg). LCMS (ESI) [M+H] + =696.2; 1H NMR (400MHz, DMSO-d6) δ12.85(s,1H),8.96(s,1H),7.86(s,1H),7.77(d,J=7.8Hz,1H),7.67(d,J=7.7Hz,1 H),7.58(t,J=7.8Hz,1H),7.52–7.35(m,1H),6.98–6.80(m,1H),6.49–6.34(m,1H),6.01(s,2H),5.59(s,1H ),5.34(s,1H),4.89–4.49(m,2H),3.69–3.37(m,2H),3.27(d,J=4.5Hz,1H),3.14(d,J=1.6Hz,3H),3.05–2 .98(m,1H),2.73–2.66(m,1H),1.91–1.72(m,2H),1.52–1.39(m,7H),0.96–0.90(m,2H),0.76–0.69(m,1H).

[0367] Example 29, Example 30

[0368] Preparation of (S)-2-((R)-2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-71) and (S)-2-((S)-2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-72):

[0369] (2S)-2-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)benzoyl)-5,7-dichloro-1-ethyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamido)-3-(3-(methanesulfonyl)phenyl)propionic acid (II-70, 153 mg) was passed through SFC (15 30% EtOH DEA C2AD) to give: II-70-P1 (II-71 or II-72) (48.61 mg). LCMS(ESI) [M+H-18] + =676.2; Chiral separation peak time: 1.628 minutes; Chiral purity 100%; 1H NMR (400MHz, DMSO-d6) δ12.86(s,1H),9.06–8.91(m,1H),7.86(s,1H),7.77(d,J=7.7Hz,1H),7.67(d,J=7 .4Hz,1H),7.60–7.41(m,2H),7.03–6.80(m,1H),6.49–6.33(m,1H),6.01(s,2H),5.57(d,J=5.6Hz,1H),5 .34(s,1H),4.86–4.41(m,2H),3.76–3.37(m,2H),3.27(d,J=4.4Hz,1H),3.15(s,3H),3.01(dd,J=14.0,1 0.4Hz,1H),2.86–2.68(m,1H),1.92–1.70(m,2H),1.46(s,6H),0.92(t,J=6.7Hz,2H),0.77–0.65(m,1H).

[0370] II-70-P2 (II-71 or II-72) (43.22 mg). LCMS(ESI)[M+H-18] + =676.2; Chiral separation peak time: 4.627 minutes, chiral purity 98.16%; 1 H NMR (400MHz, DMSO-d6) δ12.85(s,1H),9.07–8.96(m,1H),7.86(s,1H),7.77(d,J=7.8Hz,1H),7.67(d,J=7.8Hz ,1H),7.58(d,J=7.7Hz,1H),7.52–7.35(m,1H),6.97–6.83(m,1H),6.47–6.32(m,1H),6.01(s,2H),5.58(d,J= 6.1Hz,1H),5.34(s,1H),4.86–4.48(m,2H),3.71–3.35(m,2H),3.27(d,J=4.5Hz,1H),3.14(s,3H),3.04–2.98 (m,1H),2.86–2.69(m,1H),1.88–1.69(m,2H),1.47(d,J=4.8Hz,6H),0.92(t,J=6.7Hz,2H),0.77–0.68(m,1H).

[0371] Split conditions: Instrument system: Waters SFC 150; Column name: Column specifications: 250*30mm 10μm; Mobile phase A: supercritical carbon dioxide; Mobile phase B: ethanol (+0.1% 7.0M ammonia-methanol); Mobile phase ratio A:B = 65:35; Detection wavelength: 214nm; Flow rate: 140ml / min; Column temperature: room temperature; Back pressure: 100bar; Injection volume: 1mL; Cycle time: 6min; Solvents: ethanol (preparative grade), supercritical carbon dioxide (food grade); Sample preparation: The sample was dissolved in 20mL of ethanol.

[0372] The following compounds were synthesized according to Examples 1-30:

[0373] Biological Experiment Examples

[0374] Experimental Example 1: Aldehyde Complexation Reaction

[0375] Test 1:

[0376] 1.1 Reagents and Instruments

[0377] The main reagents include 4-hydroxynonenal (Medix / EN1326-012), trioleic acid glyceride (Adamas), and linoleic acid (Adamas).

[0378] The main instruments used in the experiment included a magnetic stirrer (Mei Yingpu / 98-2) and a high-performance liquid chromatography (Agilent / 1260).

[0379] 1.2 Experimental Methods

[0380] 1) Preparation of reagents for the 4-hydroxynonenal complexation reaction:

[0381] a) Reference compound ADX 102 Solution preparation: Accurately weigh compound ADX 102 and dissolve it in dimethyl sulfoxide to prepare two solutions.

[0382] b) Preparation of test compound solutions: Accurately weigh the test compound and dissolve it in dimethyl sulfoxide to prepare two solutions.

[0383] c) Preparation of reaction solution: Add 4-hydroxynonenal (5.00 mg, 32.01 μmol), trioleic acid glyceride (300 mg, 80% purity), DuPont buffer (1.25 mL, 1X), and linoleic acid (300 mg) to an 8 mL reaction flask. Prepare four solutions, divided into two groups. Add ADX 102 solution and the test compound solution to each group. Start stirring with a magnetic stirrer (20°C) and start timing. Take samples at 10, 100, 200, and 300 minutes for HPLC analysis (pipette samples of 25 μL of the upper emulsion layer and 50 μL of the aqueous phase, respectively, then dilute with 1 mL of methanol and filter). The result is the percentage content of the complexed product at 254 nm.

[0384] The compounds of this application have the ability to capture toxic aldehydes, and the results of exemplary compounds are shown in Table 1.

[0385] Table 1: Complexation reaction of 4-hydroxynonenal with the test compound and ADX102

[0386] Test 2:

[0387] 1.1 Reagents and Instruments

[0388] The main reagents include 4-hydroxynonenal (Biode) and DuPont buffer.

[0389] The main instruments used in the experiment included a magnetic stirrer (Mei Yingpu / 98-2) and a liquid chromatography-mass spectrometry (LC-MS) system (Waters).

[0390] 1.2 Experimental Methods

[0391] 1) Preparation of reagents for the 4-hydroxynonenal complexation reaction:

[0392] a) Preparation of the reference compound ADX102 solution: Accurately weigh compound ADX102 and place it in a 4 ml single-necked bottle.

[0393] b) Preparation of test compound solutions: Accurately weigh the test compounds and place them in 4 ml single-necked bottles.

[0394] c) Preparation of 4-hydroxynonenal reaction solution: Accurately weigh 4-hydroxynonenal (7.5 mg, 48.00 μmol) and dissolve it in dimethyl sulfoxide (0.15 mL) to prepare a solution. Divide the solution into 3 equal portions, each 0.05 mL, for later use.

[0395] Add 0.75 mL of DuPont buffer (1.0X) to each of the single-necked flasks containing ADX102 and the test compound. Then add the prepared 4-hydroxynonenal solution, stir with a magnetic stirrer (30 °C) and start timing. Take samples at 10, 100, 200, and 300 minutes for LC-MS analysis (take 50 μL of the reaction solution with stirring using a pipette, then dilute with 1 mL of methanol and filter). The result is the percentage content of the complexed product at 254 nm.

[0396] The compounds of this application have the ability to capture toxic aldehydes; the results for exemplary compounds are shown in Table 2.

[0397] Table 2: Complexation reactions of 4-hydroxynonenal with the test compound and ADX102

[0398] Test 3:

[0399] 1.1 Reagents and Instruments

[0400] The main reagents include malondialdehyde aqueous solution (Biode) and DuPont buffer (1X).

[0401] The main instruments used in the experiment included a magnetic stirrer (Mei Yingpu / 98-2) and a liquid chromatography-mass spectrometry (LC-MS) system (Waters).

[0402] 1.2 Experimental Methods

[0403] 1) Preparation of malondialdehyde reaction reagent:

[0404] a) Reference compound ADX102: Accurately weigh compound ADX102 into a 4 ml single-necked flask.

[0405] b) Test compound: Accurately weigh the test compound and place it in a 4 ml single-necked bottle.

[0406] c) Preparation of reaction solution: Add malondialdehyde aqueous solution (1.0 ml, 1.0 mg / ml) and DuPont buffer (0.5 ml, 1X) to the single-necked flasks containing ADX102 and the test flasks, respectively. Start stirring with a magnetic stirrer (30°C) and start timing. Take samples at 10 minutes, 100 minutes, 200 minutes, and 300 minutes for LCMS analysis (take 50 μL of reaction solution with stirring using a pipette, then dilute with 1 mL of methanol and filter). The result is the percentage content of the complexed product at 254 nm.

[0407] The compounds of this application have the ability to capture toxic aldehydes; the results for exemplary compounds are shown in Table 3.

[0408] Table 3: Complexation reactions of malondialdehyde with the test compounds and ADX102

[0409] Experimental Example 2: Competitive assay of antagonists in LFA-1 / ICAM-1 and LFA-1 / small molecule ELISA

[0410] 2.1 Reagents and Instruments

[0411] 2.2 Experimental Methods:

[0412] Human recombinant ICAM-1 / CD54 protein was prepared to a final concentration of 1 μg / mL using TBS buffer. 50 μL of the solution was transferred to a 96-well plate and incubated overnight at 4°C. The plate was washed three times with buffer, and then 150 μL of blocking buffer was added, with incubation at 37°C for 1 h. The plate was washed three times with buffer, and 1 μg / mL of human integrin αL / β2 dimer protein was prepared using buffer containing 0.1% BSA. 50 μL of the integrin protein was transferred to a 96-well plate, and 1 μL of different concentrations of the compound or DMSO was added, with incubation at room temperature for 2 h. 1 μg / mL of human integrin β2 / CD18 biotinylated antibody was prepared using buffer containing 0.1% BSA. The plate was washed three times with buffer, and 50 μL of the antibody was added, with incubation at room temperature for 1 h. The plate was washed three times with buffer, and 50 μL of Streptavidin-HRP was added, with incubation at room temperature for 20 min. Wash the plate three times with buffer, add 50 μL of TMB substrate, and incubate at room temperature for 20 min. Finally, add 25 μL of stop buffer and read the OD value of the plate at 450 nm using a microplate reader. Calculate the IC50 using GraphPad Prism 5 software. 50 Value. The compounds of this application can effectively inhibit the binding of LFA-1 and ICAM-1. The results for exemplary compounds are shown in Table 4 below.

[0413] Table 4: Results of Competitive Binding Experiments of Compounds

[0414] Note: A: less than or equal to 200 nM.

[0415] Experiment Example 3: Jurkat Cell Adhesion Inhibition Assay

[0416] 3.1 Reagents and Instruments

[0417] 3.2 Experimental Methods:

[0418] The ligand, recombinant human ICAM 1 / CD54 protein (R&D, catalog number 720-IC), was prepared to a concentration of 5 μg / mL using Tris-Buffered Saline buffer (Boston BioProducts, catalog number BM-300). 50 μL of this solution was transferred to a 96-well plate (Corning, catalog number 3690) and incubated overnight at 4°C. The plate was washed three times with Tris-Buffered Saline buffer, and then 150 μL of 0.1% BSA blocking buffer (BSA dissolved in Tris-Buffered Saline buffer) was added. Blocking was performed at 37°C for 1 h. The plate was washed three times with Tris-Buffered Saline buffer, and Jurkat cells (ATCC, catalog number TIB 152) were resuspended in Tris-Buffered Saline buffer. 200,000 cells / 50 μL were transferred to a 96-well plate, and 1 μL of different concentrations of the compound or dimethyl sulfoxide (Sigma, catalog number 34943) was added. The plate was incubated at room temperature for 2 h. Remove the liquid from the 96-well plate, wash the plate twice with Tris-Buffered Saline, add 50 μL of substrate (4-nitrophenyl-N-acetyl-β-D-glucosamine) (TCI, catalog number N0866), and incubate at 37°C for 2 h. Add 90 μL of stop buffer, and read the OD value of the plate at 405 nm using a FlexStation3 (Molecular Devices) microplate reader. Calculate the IC50 using GraphPad Prism 5 software. 50 value.

[0419] Table 5: Results of Compound Cell Adhesion Experiment

[0420] The IC50 inhibitory effect of the compound on cell adhesion assay 50 Value: A≤20nM, 20nM <B≤50nM,50nM<C≤200nM.

[0421] Experiment Example 4: Study on the Distribution of Compounds in Rabbit Eye Tissue

[0422] 4.1 Animal Information: Female New Zealand White Rabbit weighing 2-3kg, purchased from Jiashan Golden Rabbit Industry Professional Cooperative.

[0423] 4.2 Preparation of the test compound: Weigh 17 mg of test compound II-24 and add it to 0.9 mL of sterile physiological saline. Add 7.30 mg of NaOH, vortex to mix, then add saturated aqueous solution of sodium dihydrogen phosphate to adjust the pH to between 6.75 and 7.25. Finally, add sterile physiological saline to a total volume of 1 mL. A solution containing test compound II-24 with a concentration of 17 mg / mL is obtained. Prepare fresh before use.

[0424] 4.3 Sample Collection

[0425] After administering eye drops (50 μL per eye) to both eyes of 6 animals, tissue and blood samples were collected at 0.5 and 2.0 hours. Approximately 500 μL of blood sample was collected and placed in an anticoagulant tube containing EDTA-K2 anticoagulant, and centrifuged within 60 minutes to obtain plasma. Whole blood samples were placed on wet ice before centrifugation. A total of 6 plasma samples were collected. After plasma collection, the animals were euthanized by CO2 inhalation (3 animals each at 0.5 and 2 hours). Local ocular tissues were collected from both eyes: cornea, bulbar conjunctiva, and palpebral conjunctiva. After rinsing with physiological saline, the surface moisture was blotted with filter paper, weighed, and stored. All samples were weighed and stored separately for both sides. A total of 12 corneas, 12 bulbar conjunctivaes, and 12 palpebral conjunctivaes were collected. All collected plasma samples were stored on dry ice or frozen until analysis.

[0426] 4.4 Sample processing procedures and testing methods

[0427] Extraction process of plasma standard curve, quality control and samples (blank and test samples): Add 70.0 μL of internal standard working solution to each of the 96-well plates containing 7.0 μL of prepared Blank, standard curve, quality control and test samples respectively. Place the 96-well plate on a mixer and mix for 10 minutes (temperature set to room temperature, speed set to 500 rpm). Centrifuge for 10 minutes (temperature set to 4℃, centrifugation force set to 4000 rpm). Take 50.0 μL of supernatant into a 96-well plate containing 50.0 μL of ultrapure water. Place the 96-well plate on a mixer and mix for 10 minutes (temperature set to room temperature, speed set to 500 rpm).

[0428] Rabbit blank eyeballs, corneas, bulbar conjunctiva, and palpebral conjunctiva tissues were homogenized with 50% acetonitrile solution at a weight-to-volume ratio of 1:5 (w / v) to obtain an ocular tissue homogenate.

[0429] Extraction process of standard curve, quality control and samples (blank and test) of eye tissue homogenate: Add 300.0 μL of internal standard working solution to each of the 96-well plates containing 30.0 μL of prepared Blank, standard curve and quality control samples. Mix the 96-well plates on a mixer for 10 minutes (temperature set to room temperature, speed set to 500 rpm), centrifuge for 10 minutes (temperature set to 4℃, centrifugation force set to 4000 rpm), take 100.0 μL of supernatant into a 96-well plate containing 100.0 μL of ultrapure water, and mix the 96-well plates on a mixer for 10 minutes (temperature set to room temperature, speed set to 500 rpm).

[0430] 4.5 Test Results:

[0431] Table 6:

[0432] The experimental results indicated that after ophthalmic administration of compound II-24 to rabbits, the drug concentrations were high in the cornea, bulbar conjunctiva, and palpebral conjunctiva, but low in plasma.

[0433] Experimental Example 5: Scopolamine-induced mouse dry eye model test

[0434] 5.1 Animal Information: 8-10 week old male C57BL / 6J mice were purchased from Shanghai Jihui Laboratory Animal Breeding Co., Ltd.

[0435] 5.2 Preparation of the test compound: Weigh 5g of the test compound and add it to 90mL of sterile physiological saline. Add 0.7g of NaOH and stir to obtain a clear solution. Add saturated NaH2PO4 (sodium dihydrogen phosphate) aqueous solution to the obtained solution until the pH of the solution is between 6.75 and 7.25 (measure the pH value using accurate pH test paper or a pH meter). Add sterile physiological saline to the obtained aqueous solution until the total volume reaches 100.0mL. Aliquot the resulting solution into sterile EP tubes according to the daily usage and store at 2-8℃ protected from light. The solvent is a system without the compound. A 1% (mass fraction) solution is obtained by diluting a 5% (mass fraction) solution.

[0436] 5.3 Experimental Reagents and Instruments

[0437] Scopolamine hydrobromide (McLean), Restasis ophthalmic emulsion (Allergan Sales, LLC), fluorescein sodium injection (Alcon Laboratoryes, Inc.); handheld slit lamp (SL-17, Kowa).

[0438] 5.4 Model Creation Environment:

[0439] Mice will be housed in transparent resin-plastic cages (390mm × 180mm × 130mm) in the animal room, with 2-5 mice per cage. The cage bedding will be autoclaved corn cob bedding (from Chenfu Eden Bedding Processing Plant, Dachang Hui Autonomous County), changed every 7-10 days. The cage numbers will be recorded in the experimental log throughout the experiment. The animal room will be equipped with a high-efficiency air filter, with ventilation at 15-25 times per hour. The temperature will be maintained between 20-26℃ (68-79°F), and the relative humidity between 30-70%. Temperature and humidity will be continuously observed and recorded. Lighting conditions will consist of 12 hours of fluorescent lighting (08:00-20:00) and 12 hours of no lighting per day.

[0440] 5.5 Experimental Design

[0441] 5.5.1 Modeling

[0442] Before modeling, detailed eye observation was performed using a slit lamp. Animals with no abnormalities in both eyes were selected for the group. Six animals served as the G1 normal control group, and the remaining animals were used for modeling.

[0443] After grouping on Day 1, all animals except G1 were housed in a drying box with relative humidity controlled below 40%. Animals G2–G8 were subcutaneously injected with 0.25% scopolamine hydrobromide (SCO) to induce the scopolamine model, while animals G1 were subcutaneously injected with normal saline (NS) as a normal control group. The dosage was 0.2 mL / time, three times a day (time points: 9:00, 13:00, 17:00, ±1h) for 14 days.

[0444] 5.5.2 Grouping and Administration

[0445] Before administration, animals were grouped into relatively uniform groups based on their tear secretion volume.

[0446] Groups 1-8: Dosing from Day 1 to Day 13, eye drops, twice daily, 15 μL / eye / time (once an eye drop only on the morning of Day 14).

[0447] 5.5.3 Detection Indicators

[0448] Schirmer I test (SIT)

[0449] On Day 14 after modeling, all enrolled animals underwent the Schirmer I test to examine tear secretion. Mice were immobilized, and a phenol-red cotton thread was inserted into the temporal side of the lower eyelid conjunctival sac using micro-forceps. After 30 seconds, the thread was removed, and the length of the discolored thread after wetting was measured with calipers, accurate to 0.01 mm. If the thread fell out, a new thread was used after a 5-minute interval, and the measurement was repeated.

[0450] Corneal fluorescence staining (CFS) score

[0451] On Day 14 after modeling, all enrolled animals underwent CFS scoring to assess ocular surface damage. Mice were immobilized, and 20 μL of 0.5% sodium fluorescein solution was instilled into the superior conjunctival sac. Blinking was assisted to ensure even distribution of the sodium fluorescein across the cornea, followed by rinsing with physiological saline. Observation was performed under slit-lamp cobalt blue light, and the fluorescein staining was evaluated using a 16-point scale. The corneal center was used as the origin, and the cornea was divided into four quadrants. The scoring criteria for each quadrant are shown in Table 7, with each animal scoring 0-16 points per eye.

[0452] Table 7. CFS Scoring Criteria

[0453] 5.6 Statistical Analysis

[0454] Data are presented as mean ± standard error. Statistical analysis was performed using Graphpad Prism, SPSS, or Sigmaplot software. Detailed data are presented in graphical form. A p-value < 0.05 was considered statistically significant.

[0455] Experimental results showed that in the scopolamine-induced dry eye model in mice, the above-mentioned preferred compounds significantly improved both tear secretion and corneal fluorescence staining indicators.

Claims

1. A compound represented by formula (Va), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound: in, Ring A is a 5-12 membered heteroaryl or a C6-12 aryl; Ring B is a 5-12 membered heteroaryl or a C6-12 aryl; R A Each of the following is independently selected from deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more elements selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups; R B Each of the following groups is independently selected from deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups; a is selected from 0, 1, 2, 3, 4, 5, or 6; b is selected from 1, 2, 3, 4, 5, or 6; R S Each element is independently selected from deuterium, halogens, -CN, -OH, -NH2, and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkyl thio group; or 2 R groups S Together with the atoms it is attached to, they form C 3-8 cycloalkyl; n is 0, 1, 2, 3 or 4; R is independently selected from hydrogen, halogen, or C. 1-6 alkyl; Unless otherwise stated, the heteroatoms in the above heteroaryl groups are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4; The condition is that, In this case, 'a' must be at least 2, and there are two 'R's. A They are -NH2 and And adjacent, and / or, In the case of b, it is at least 2, and there are two R values. B They are -NH2 and And they are adjacent.

2. A compound represented by formula (I'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound: in, Ring A is a 5-12 membered heteroaryl or a C6-12 aryl; Ring B is a 5-12 membered heteroaryl or a C6-12 aryl; R A For deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups; R B For deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, or optionally substituted: C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups; a is selected from 0, 1, 2, 3, 4, 5, or 6; b is selected from 1, 2, 3, 4, 5, or 6; R is independently selected from hydrogen, halogen, or C. 1-6 alkyl; Unless otherwise stated, the heteroatoms in the above heteroaryl groups are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4; The condition is that, Include Structure, and / or Include Structure; dashed lines represent single or double bonds.

3. The compound according to any one of claims 1-2, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: Ring A is a 6-10 membered heteroaryl or phenyl group; Alternatively, ring A can be pyridyl, quinolinyl, phenyl, or benzofuranyl.

4. The compound according to any one of claims 1-3, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: R A Each of the following is independently selected from deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more groups selected from deuterium, -OH, -SH, -NH2, C 1-4 Substituents of alkyl groups; Or, R A C is independently selected from halogens, -OH, -NH2, or optionally substituted: 1-6 Alkyl, C 3-6 Cycloalkyl, C1-6 alkoxy; wherein optional substitution means unsubstituted or substituted by one or more substituents selected from halogens and -OH; Or, R A -NH2 or 5. The compound according to any one of claims 1-4, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: a is selected from 1, 2, 3, 4, 5 or 6; or a is selected from 0, 1, 2 or 3; or a is selected from 0, 2, 3 or 4; or a is selected from 1 or 2; or a is 2.

6. The compound according to any one of claims 1-5, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: a is 2, and both R A Located adjacent; or, a is 2, and both R are adjacent. A Located adjacent, two R A They are -NH2 and Alternatively, a is 3, and two of the R values ​​are... A Located adjacent, two R A They are -NH2 and Another R A Selected from deuterium, halogens, -CN, -OH, -NH2, C 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio; Alternatively, a is 3, and two of the R values ​​are... A Located adjacent, two R A They are -NH2 and Another R A Selected from F, Cl, -OH, methyl, methoxy, cyclopropyl, or trifluoromethyl; Alternatively, a is 4, and two of them are R. A Located adjacent, two R A They are -NH2 and The other two R A Selected from halogens.

7. The compound according to any one of claims 1-6, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: for X1, X2, or X3 are each independently selected from CH and CR. A Or N; or, for X1, X2, or X3 are each independently selected from CH and CR. A Or N; where each R A Independently deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 Cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted with one or more radicals selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 Alkyl substituents are substituted; or, each R A Independently, it is deuterium, halogen, -CN, -OH, -NH2, methyl, trifluoromethyl, methoxy, cyclopropyl; or, each R A Independently, it can be deuterium, halogen, -CN, -OH, -NH2; or, each R A For F; or, for X1, X2, or X3 are each independently selected from CH and CR. A Or N; or one of X1, X2, or X3 is N, and the other two are CH; or one of X1, X2, or X3 is N, and the other two are CH and CR respectively. A Alternatively, one of X1, X2, or X3 is N, and the other two are CR. A ; or X1, X2, or X3 are all CH; or one of X1, X2, or X3 is CR. A The other two are both CH; or one of X1, X2, or X3 is CH, and the other two are both CR. A ; where each R A Independently deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted by one or more elements selected from deuterium, halogen, -OH, -SH, -NH2, C 1-4 Alkyl substituents are substituted; or, each R A Independently halogenated, -OH, or optionally substituted: C 1-6 Alkyl, C 3-6 cycloalkyl, C 1-6 alkoxy; wherein optional substitution means unsubstituted or substituted with one or more halogens; or, each R A Independently F, Cl, -OH, methyl, trifluoromethyl, methoxy, cyclopropyl; or, each R A For F; or, for or, for 8. The compound according to any one of claims 1-7, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: Ring B is a 6-10 membered heteroaryl or phenyl group; Alternatively, ring B can be pyridyl or phenyl.

9. The compound according to any one of claims 1-8, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: R B -NH2, methanesulfonyl or Alternatively, b can be selected from 1 or 2; or, when b is 1, R B It is methanesulfonyl; when b is 2, R B They are -NH2 and Or, when b is 1, R B It is methanesulfonyl; when b is 2, R B They are -NH2 and And two Rs B Adjacent; or, for 10. The compound according to any one of claims 1 or 3-9, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: R S Selected independently from deuterium, halogens, and C 1-4 Alkyl, C 1-4 Haloalkyl, C 3-6 cycloalkyl or C 1-4 alkoxy group; or 2 R groups S Together with the atoms it is attached to, they form C 3-6 cycloalkyl; Or, R S Selected independently from deuterium, halogens, and C 1-4 Alkyl or C 1-4 Alkoxy; Or, R S Each of the following is independently selected from deuterium, F, methyl, ethyl, and methoxy.

11. The compound according to any one of claims 1 or 3-10, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: n is 0, 1, or 2; Alternatively, n can be 0 or 1.

12. The compound according to any one of claims 1 or 3-11, a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein: R is independently selected from hydrogen or halogen; or, R is independently selected from hydrogen or Cl; or, the two Rs are hydrogen and Cl respectively; or, the two Rs are both Cl.

13. A compound represented by formula (II') or (III'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound: in, Ring A, R A a, R are as described in any one of formula (Va) or formula (I') of claims 1-12; Among them, ring A, ring B, R B b, R are as described in any one of the formulas (Va) or (I') of claims 1-12.

14. A compound represented by formula (V-a'), formula (V), formula (VI), formula (VI-a), formula (VII), or formula (VII-a), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound: in, Ring A, R A Ring B, R B R S a, b, n, R are as described in any one of the formulas (Va) of claims 1 or 3-12; Among them, ring A, ring B, R A R B R S a, b, and n are as described in any one of the compounds of formula (Va) as claimed in claim 1 or 3-12; Among them, X1, X2, or X3 are independently selected from CH and CR, respectively. A Or N; ring B, R A R B b, R, R S n is the compound of any one of formula (Va) as claimed in claim 1 or 3-12; Among them, rings B and R A R B b, R, R S n is the compound of any one of formula (Va) as claimed in claim 1 or 3-12; X1, X2, or X3 are each independently selected from CH and CR. A Or N; R A R S n is the compound of any one of formula (Va) as claimed in claim 1 or 3-12; where R A R S The compound of any one of formula (Va) as claimed in claim 1 or 3-12.

15. A compound, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

16. A compound, a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

17. A pharmaceutical composition comprising the compound of any one of claims 1-16, a stereoisomer, tautomer, or mixture thereof of the compound, a pharmaceutically acceptable salt of the compound, and optionally, further comprising a pharmaceutically acceptable carrier.

18. Use of the compound of any one of claims 1-16, a stereoisomer, tautomer, or mixture thereof, a pharmaceutically acceptable salt of the compound, or the pharmaceutical composition of claim 17 in the preparation of a medicament for treating and / or preventing LFA-1-mediated diseases and / or diseases involving aldehyde toxicity.

19. The use as described in claim 18, wherein, The LFA-1-mediated and / or pathogenesis involving aldehyde toxicity is an ocular disease; preferably, the ocular disease is a chronic ocular surface disease; preferably, the ocular disease is dry eye syndrome.

20. An intermediate for preparing the compound represented by formula (VI-a): in, X1, X2, X3, R S Or n as described in compound (VI-a) of formula 14.

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