Class of benzoylglycine derivatives, preparation method therefor and use thereof

Abstract

Provided are a compound serving as an LFA-1 antagonist and / or a RASP scavenger, and a stereoisomer and a tautomer thereof, or a mixture thereof, a pharmaceutically acceptable salt thereof, a method of preparing the compound, and the use thereof in treating LFA-1 and / or RASP. In-vitro activity test results show that the compound has an excellent ability to capture toxic aldehydes and the ability to inhibit the binding of LFA-1 to ICAM-1.

Description

Benzoyl glycine derivatives and methods of making and using the same

[0001] Cross-reference to Related Applications

[0002] This application claims priority to and the benefit of Chinese Patent Application No. 202411248025.3, filed September 6, 2024, and Chinese Patent Application No. 202411679367.0, filed November 22, 2024, in the China National Intellectual Property Office, the disclosures of which are incorporated herein in their entireties by reference. TECHNICAL FIELD

[0003] The present application relates to the technical field of medicine, in particular to a class of benzoyl glycine derivatives, and a preparation method and use of the compounds. BACKGROUND

[0004] Dry eye disease, also known as keratoconjunctival xerosis, is a refractory ocular surface disease, and the main symptoms include ocular dryness, foreign body sensation, burning sensation, etc. Dry eye disease is a chronic ocular surface disease caused by multiple factors, and the medical community generally believes that the main pathological mechanism is that the instability of the tear film or the imbalance of the ocular surface microenvironment caused by the abnormality of the quality or quantity of the tear film, accompanied by ocular surface inflammatory response, tissue damage and abnormal nerve sensation. Dry eye disease, if not diagnosed and treated in time, can lead to further complications such as infection, ocular surface keratinization, corneal ulcer and conjunctival squamous metaplasia, etc.

[0005] Dry eye disease is a continuous pathological process, and the disease condition continuously develops from mild to severe, and there is no obvious boundary between mild, moderate and severe. At present, the treatment of dry eye disease mainly includes three kinds of therapy, i.e. drug, physical and surgical treatment.

[0006] Although the etiology of dry eye is complex, researchers have found that the pathology is similar: immune cell infiltration into the surface tissue of the eye triggers chronic inflammation, causing ocular surface damage, and anti-inflammatory treatment has become the direction of dry eye drug development. There are two main anti-inflammatory directions in the current European and American markets: immunosuppressants and active aldehyde capture agents. There are two drugs approved for immunosuppressants: (1) cyclosporine A suspension, which is a suspension, has problems of long-term instability and suspension irritation to the eye, and as a strong immune system inhibitor, it can cause damage to the immune system. (2) Lifitigrast, 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 efficacy, and was approved by FDA in December 2016, but has high lipophilicity, long-term use can cause taste degeneration, and more than 50% of patients have no effect in clinical practice. The fastest drug for active aldehyde capture agent is ADX-102. RASP (reactive aldehyde) covalently reacts with amine and thiol groups. After binding to a threshold amount of electrophilic reagent-responsive 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-kB; in addition, RASP protein adducts bind to scavenger receptor A, which initiates pro-inflammatory signals and forms antibodies against adduct proteins, thereby enhancing the inflammatory response. ADX-102 can capture RASP, thereby blocking the subsequent series of reactions mediated by it, achieving an anti-inflammatory effect, and thus achieving the purpose of treating dry eye, but ADX-102 has poor water solubility, and with the help of excipients, the highest concentration of eye drops is only 0.5%, which may be due to insufficient efficacy and too much eye irritation. At present, it is still in the supplementary clinical phase III study. SUMMARY

[0007] The present application aims to provide a novel compound as an LFA-1 antagonist and / or RASP capture agent, a preparation method of the compound and its use in treating eye diseases (e.g., dry eye).

[0008] In a first aspect of the present application, a compound represented by the following formula (A) is provided, a stereoisomer, a tautomer or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound:

[0009] wherein,

[0010] L is or optionally substituted: C 1-6 alkylene, C 2- 6alkenylene, C 2-6 alkynylene, C 1-6alkeneoxy, C 1-6 Alkylthio group; wherein optional substitution means unsubstituted or substituted with one or more groups selected from deuterium, halogen, oxo group, -OH, -SH, -NH2, C 1-4 Alkyl substituents; * indicates the A-ring junction; n is 1, 2, or 3;

[0011] Z1, Z2, Z3, and Z4 are each independently selected from N or CR. Z ;

[0012] R Z Each of the following is independently hydrogen, deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 3-6 Cycloalkyl; wherein optional substitution means unsubstituted or substituted by one or more substituents selected from deuterium, halogen, -OH, -SH, -NH2;

[0013] Ring A is a 5-12 membered heteroaryl group or C 6-12 Aryl;

[0014] Ring B is a 5-12 membered heteroaryl group or C 6-12 Aryl;

[0015] R A Each can be independently deuterium, halogen, -CN, -OH, -NH2, or optionally substituted: C 1-6 Alkyl, C 2-6 alkenyl, 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, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-6 Substituents of cycloalkyl groups;

[0016] R B Each can be independently deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, -S(O)2NH2, -P(O)(CH3)2, -P(O)2CH3, -S(O)(NH)CH3, 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;

[0017] Rc is selected from hydrogen, deuterium, and C.1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl;

[0018] Rd is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl;

[0019] R f1 and R f2 are each independently selected from hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, C 1- 6alkoxy, C 1-6 alkylthio; or R f1 and R f2 together with the C atom to which they are attached form a C 3-8 cycloalkyl;

[0020] R1 and R2 are each independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl;

[0021] a is selected from 2, 3, 4, 5 or 6;

[0022] b is selected from 1, 2, 3, 4, 5 or 6;

[0023] Unless otherwise specified, the heteroatoms in the above-mentioned heteroaryl, heterocyclyl are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4;

[0024] provided that, two R A are -NH2 and -NH2 is adjacent to .

[0025] In some embodiments of the application, n is 1 or 2. In some embodiments of the application, n is 1.

[0026] In some embodiments of the application, L is or optionally substituted: C 1-3 alkylene, C 2-6 alkynylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from deuterium, halogen, oxo, -OH, -SH, -NH2, C 1-4 alkyl; * indicates the point of attachment of ring A;

[0027] Z1, Z2, Z3 and Z4 are each independently selected from N or CRZ ;

[0028] R Z is hydrogen, deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-3 alkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2;

[0029] Rcis selected from the group consisting of hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl;

[0030] R f1 and R f2 are each independently selected from the group consisting of hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-3 alkyl, C 1-3 haloalkyl, C 3-6 cycloalkyl; or R f1 and R f2 together with the C atom to which they are attached form a C 3-6 cycloalkyl.

[0031] In some embodiments of the present application, L is or optionally substituted C 1-3 alkylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2; * indicates the ring A attachment end;

[0032] one, two or three of Z1, Z2, Z3and Z4are N, and the others are CH;

[0033] Rcis selected from the group consisting of hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl;

[0034] R f1 and R f2 are each independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-3 alkyl; or R f1 and R f2 together with the C atom to which they are attached form a C 3-6 cycloalkyl.

[0035] In some embodiments of the present application, L is * indicates the ring A attachment end;

[0036] Rcis selected from the group consisting of hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl;

[0037] R f1 and R f2independently selected from the group consisting of hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-3 alkyl; or R f1 and R f2 form, together with the C atom to which they are attached, a C 3-6 cycloalkyl.

[0038] In some embodiments of the application, L is * indicates the ring A attachment point; wherein Rcis hydrogen, methyl, ethyl or cyclopropyl; R f1 and R f2 are independently selected from the group consisting of hydrogen, deuterium and methyl (e.g., R f1 and R f2 are both hydrogen, or both deuterium, or both methyl, or one is methyl and the other is hydrogen); or R f1 and R f2 form, together with the C atom to which they are attached, a cyclopropyl.

[0039] In some embodiments of the application, L is * indicates the ring A attachment point.

[0040] In some embodiments of the application, L is -CONH(CH2)2-.

[0041] In some embodiments of the application, L is or optionally substituted C 1-3 alkylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, oxo, -OH, -SH, -NH2, C 1-4 alkyl; * indicates the ring A attachment point;

[0042] one, two or three of Z1, Z2, Z3and Z4are N, and the others are CR Z (or the others are CH);

[0043] R Z is hydrogen, deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-3 alkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -NH2.

[0044] In some embodiments of the application, L is * indicates the ring A attachment point.

[0045] In some embodiments of the application, L is -CH2-.

[0046] In some embodiments of the application, L is In some embodiments of the application, L is In some embodiments of the application, L is

[0047] In some embodiments of the application, ring A is 5-10 membered heteroaryl or phenyl.

[0048] In some embodiments of the application, ring A is thienyl, quinolinyl or phenyl.

[0049] In some embodiments of the application, ring A is phenyl.

[0050] In some embodiments of the application, a is 2, 3 or 4, wherein two R A are -NH2 and respectively, and are adjacent.

[0051] In some embodiments of the application, a is selected from 2 or 3.

[0052] In some embodiments of the application, a is 2.

[0053] In some embodiments of the application, a is 2, and two R A are located in the ortho position.

[0054] In some embodiments of the application, a is 2, and two R A are located in the ortho position, and two R A are -NH2 and

[0055] In some embodiments of the application, a is 3, and wherein two R A are located in the ortho position, and two R A are -NH2 and the other R A is selected from deuterium, halogen, -CN, -OH, -NH2, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio.

[0056] In some embodiments of the application, a is 3, and wherein two R A are located in the ortho position, and two R A are -NH2 and the other R A is selected from halogen (e.g. F) or -OH.

[0057] In some embodiments of the application, is

[0058] In some embodiments of the application, As In some embodiments of the application, As

[0059] In some embodiments of the application, ring B is 6-10 membered heteroaryl or phenyl.

[0060] In some embodiments of the application, ring B is phenyl.

[0061] In some embodiments of the application, R B is -NH2, methylsulfonyl, -S(O)2NH2, -P(O)(CH3)2, -P(O)2CH3, -S(O)(NH)CH3, or

[0062] In some embodiments of the application, b is 1 and R B is methylsulfonyl, -S(O)2NH2, -P(O)(CH3)2, -S(O)(NH)CH3.

[0063] In some embodiments of the application, b is 1 and R B is methylsulfonyl.

[0064] In some embodiments of the application, As In some embodiments of the application, As In some embodiments of the application, As

[0065] In some embodiments of the application, Rdis hydrogen.

[0066] In some embodiments of the application, R1and R2are each independently selected from hydrogen, halogen.

[0067] In some embodiments of the application, R1and R2are each independently selected from hydrogen, halogen, and wherein at least one is halogen.

[0068] In some embodiments of the application, R1and R2are each independently selected from hydrogen, Cl; and wherein at least one is Cl.

[0069] In some embodiments of the application, R1and R2are both halogen. In some embodiments of the application, R1and R2are both CI.

[0070] In some embodiments of the application, Rdis hydrogen and R1and R2are both CI.

[0071] In some embodiments of the application, the C atom attached to COOH is in the (S) configuration.

[0072] In some embodiments, the compound of formula (A) is a compound of formula (A') or (A''),

[0073] wherein ring A, ring B, R1, R2, R A , a, R B , b, R d , L are as described for a compound of formula (A).

[0074] In some embodiments, the compound of formula (A) is a compound of formula (I-a),

[0075] wherein,

[0076] ring A is a 5-12 membered heteroaryl or C 6-12 aryl;

[0077] ring B is a 5-12 membered heteroaryl or C 6-12 aryl;

[0078] R A is deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2, C 1-4 alkyl;

[0079] R B is deuterium, halogen, -CN, -NH2, methylsulfonyl, ethylsulfonyl, -S(O)2NH2, -P(O)(CH3)2, -P(O)2CH3, -S(O)(NH)CH3or is optionally substituted: C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, -OH, -SH, -NH2, C 1-4Substituents of alkyl groups;

[0080] Rc and Rd are each independently selected from hydrogen and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl;

[0081] Alternatively, Rc and Rd, together with their respective attached N and C atoms, form a nitrogen-containing 6-membered heterocyclic group; wherein the nitrogen-containing 6-membered heterocycle is optionally substituted by one or more Re atoms;

[0082] Each Re is independently selected from deuterium, halogen, -CN, -OH, -NH2, C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 cycloalkyl, C 1-6 Alkoxy, C 1-6 Alkylthio group; or two Re atoms together with their respective bonded atoms to form a C group. 3-8 cycloalkyl;

[0083] R f1 and R f2 Each element is independently selected from hydrogen, deuterium, halogens, -CN, -OH, -NH2, and C. 1-6 Alkyl, C 1-6 Haloalkyl, C 3-8 Cycloalkyl, C1-6 alkoxy, C 1-6 alkylthio; or R f1 and R f2 Together with the atoms it is attached to, they form C 3-8 Cycloalkyl; R1 and R2 are selected from hydrogen, halogen, and C, respectively. 1-6 Alkyl, C 1-6 Halogenated alkyl groups;

[0084] a is selected from 0, 1, 2, 3, 4, 5, or 6;

[0085] b is selected from 1, 2, 3, 4, 5, or 6;

[0086] Unless otherwise stated, the heteroatoms in the above heteroaryl and heterocyclic groups are independently selected from O, N or S, and the number of heteroatoms is 1, 2, 3 or 4;

[0087] The condition is that, Include Structure; dashed lines represent single or double bonds.

[0088] In some implementation schemes, R B For deuterium, halogen, -CN, -NH2, methanesulfonyl, ethanesulfonyl, or optionally substituted: C 1-6 Alkyl, C 3-8 cycloalkyl, C 1-6Alkoxy, 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.

[0089] In some embodiments, the compound represented by formula (Ia), R f1 and R f2 Both are hydrogen.

[0090] In some embodiments, rings A, B, R1, R2, and R in the compound represented by formula (Ia) A a, R B b, R f1 R f2 R c R d As described in compound (A).

[0091] In some embodiments, the compound represented by formula (A) is the same as the compound represented by formula (Ib).

[0092] Among them, ring A, ring B, R1, R2, R A a, R B b, R f1 R f2 R c R d As described in compounds of formula (A) or (Ia).

[0093] In some embodiments, the compound represented by formula (A) is a compound represented by formula (Ic).

[0094] Among them, ring A, ring B, R1, R2, R A a, R B b, R d As described for compound (A);

[0095] Z1, Z2, Z3, and Z4 are each independently selected from N or CR. Z ;

[0096] R Z For hydrogen, deuterium, halogen, -CN, -OH, -NH2, or optional substitution: C 1-3 Alkyl; wherein optional substitution means unsubstituted or substituted by one or more substituents selected from deuterium, halogen, -NH2.

[0097] In some implementations, one, two, or three of Z1, Z2, Z3, and Z4 are N, and the rest are CR. Z(or the rest is CH);

[0098] R Z is hydrogen, deuterium, halogen, -CN, -OH, -NH2, or is optionally substituted: C 1-3 alkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -NH2.

[0099] In some embodiments, the compound of Formula (A) is a compound of Formula (I-d),

[0100] wherein ring A, ring B, R1, R2, R A , a, R B , b, R d are as described for the compound of Formula (A).

[0101] The present application also provides a compound of Formula (IV-a) as shown below, a stereoisomer, a tautomer, or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound:

[0102] wherein ring A, ring B, R1, R2, R A , a, R B , b, Rc, R f1 , and R f2 are as described for the compound of Formula (A) or Formula (I-a).

[0103] In some embodiments of the present application, Rcis selected from the group consisting of hydrogen, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl.

[0104] In some embodiments of the present application, Rcis selected from the group consisting of hydrogen, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl.

[0105] In some embodiments of the present application, Rcis selected from the group consisting of hydrogen, methyl, ethyl, cyclopropyl.

[0106] In some embodiments of the present application, Rcis selected from the group consisting of hydrogen.

[0107] In some embodiments of the present application, R f1 and R f2 are each independently selected from the group consisting of hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-6 alkyl, C 1- 6haloalkyl, C 3-8 cycloalkyl, C 1-6alkoxy, C 1-6 alkylthio; or R f1 and R f2 together with the C atom to which they are attached form a C 3-8 cycloalkyl.

[0108] In some embodiments of the application, R f1 and R f2 are each independently selected from hydrogen, deuterium, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl; or R f1 and R f2 together with the C atom to which they are attached form a C 3-6 cycloalkyl.

[0109] In some embodiments of the application, R f1 and R f2 are each independently selected from hydrogen or deuterium. In some embodiments of the application, R f1 and R f2 are each hydrogen; or are each deuterium; or are each methyl; or one is hydrogen and the other is methyl.

[0110] In some embodiments of the application, is

[0111] In some embodiments of the application, ring A is thienyl, quinolinyl or phenyl;

[0112] In some embodiments of the application, ring A is phenyl;

[0113] In some embodiments of the application, R A is -NH2or

[0114] In some embodiments of the application, a is selected from 2 or 3;

[0115] In some embodiments of the application, a is selected from 2;

[0116] In some embodiments of the application, a is selected from 2, and both R A are located in the ortho position;

[0117] In some embodiments of the application, a is selected from 2, and both R A are located in the ortho position, and both R A are -NH2and

[0118] In some embodiments of the application, a is selected from 3, and wherein two R A are located in the ortho position, and both RA 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;

[0119] In a preferred embodiment of this application, a is selected from 3, and two of R are... A Located adjacent, two R A They are -NH2 and Another R A Selected from halogens (e.g., F) or -OH.

[0120] In some embodiments of this application, for

[0121] In some embodiments of this application, R1 and R2 are selected from hydrogen and halogen, respectively, and at least one of them is a halogen.

[0122] In some embodiments of this application, R1 and R2 are selected from hydrogen and Cl, respectively; and at least one of them is Cl.

[0123] In some embodiments of this application, both R1 and R2 are Cl.

[0124] This application also provides a compound represented by formula (IV), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0125] Among them, ring A, ring B, R1, R2, R A a, R B b, as described in compounds of formula (A), (Ia), or (IV-a).

[0126] This application also provides a compound represented by formula (IV'), a stereoisomer, tautomer, or mixture thereof, or a pharmaceutically acceptable salt of the compound:

[0127] Among them, ring A, ring B, and ring R f1 R f2 R A a, R B b, R1, R2 are as described in compounds of formula (A), (Ia), or (IV-a).

[0128] The present application also provides a compound as shown in the following formula (V), a stereoisomer, a tautomer or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound:

[0129] wherein, ring A, R1, R2, R A , a is as described in the compound of formula (A), formula (I-a) or formula (IV-a).

[0130] In some embodiments of the present application, the C atom connected with COOH in formula (A), formula (I-a), formula (I-b), formula (I-c), formula (I-d), formula (IV-a), formula (IV), formula (IV’), formula (V) is (S) configuration.

[0131] In some embodiments, the present application comprises the above-mentioned definitions of variables and embodiments thereof, and any combination thereof.

[0132] The present application provides a compound, a stereoisomer, a tautomer or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound, wherein the compound is selected from:

[0133] In some embodiments, the pharmaceutical composition comprises a compound, a stereoisomer, a tautomer or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound as described in the first aspect, and one or more pharmaceutically acceptable carriers.

[0134] In some embodiments, the pharmaceutical composition comprises a compound, a stereoisomer, a tautomer or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound as described in the first aspect, and one or more pharmaceutically acceptable carriers.

[0135] In some embodiments, the pharmaceutical composition is used as an LFA-1 antagonist and / or RASP trap.

[0136] In some embodiments, the pharmaceutical composition is used for treating and / or preventing a disease mediated by LFA-1 and / or involving aldehyde toxicity in the pathogenesis, comprising a compound, a stereoisomer, a tautomer or a mixture thereof, an isotopic derivative or a pharmaceutically acceptable salt of the compound as described in the first aspect, and one or more pharmaceutically acceptable carriers.

[0137] In some embodiments, the pharmaceutical composition is for treating and / or preventing a disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity, comprising a therapeutically effective amount of the compound of the first aspect, stereoisomer, tautomer, or mixture thereof, isotopic derivative, or pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

[0138] In a third aspect, the present application provides use of the compound of the first aspect, stereoisomer, tautomer, or mixture thereof, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the second aspect in the manufacture of a medicament.

[0139] In some embodiments, the medicament is a medicament as an LFA-1 antagonist and / or RASP trap.

[0140] In some embodiments, the medicament is a medicament for treating and / or preventing a disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity.

[0141] In a fourth aspect, the present application provides the compound of the first aspect, stereoisomer, tautomer, or mixture thereof, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the second aspect for use in treating and / or preventing a disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity.

[0142] In a fifth aspect, the present application provides a method for treating and / or preventing a disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity, comprising administering to a subject in need a therapeutically effective amount of the compound of the first aspect, stereoisomer, tautomer, or mixture thereof, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the second aspect.

[0143] In a sixth aspect, the present application provides use of the compound of the first aspect, stereoisomer, tautomer, or mixture thereof, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of the second aspect in treating and / or preventing a disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity.

[0144] The disease mediated by LFA-1 and / or having an underlying mechanism involving aldehyde toxicity in the second aspect, the third aspect, the fourth aspect, the fifth aspect, and the sixth aspect is preferably an ocular disease; preferably, the ocular disease is a chronic ocular surface disease; preferably, the ocular disease is dry eye.

[0145] The technical solution or technical feature of the third aspect is applicable to the fourth aspect, the fifth aspect, and the sixth aspect in the absence of contradiction and conflict.

[0146] The compounds or pharmaceutical compositions of the present application have one or more of the following benefits: (1) the compounds of the present application have the ability to trap toxic aldehydes; (2) the compounds of the present application can effectively inhibit the binding of LFA-1 to ICAM-1; (3) the compounds of the present application have the effect of inhibiting cell adhesion; (4) after eye drop administration, the compounds of the present application have high drug concentration in the cornea, bulbar conjunctiva and palpebral conjunctiva, and low drug concentration in the plasma; (5) the compounds of the present application can significantly improve the tear secretion amount index and the corneal fluorescein staining index.

[0147] The present application also includes a method of preparing the compounds shown above, a stereoisomer, a tautomer, or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound.

[0148] The methods include, but are not limited to, the following:

[0149] General Preparation Method One:

[0150] General Preparation Method Two:

[0151] wherein, ring A, R1, R2, R A , a are as described for the compound of formula (V).

[0152] Definitions

[0153] The following terms as used herein have the following meanings, unless a different meaning is expressly provided or is apparent from the context of their use. A particular term should not be construed as indefinite or unclear in the absence of a specific definition, but should be understood in accordance with the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0154] The term "alkyl", unless otherwise specified, refers to a monovalent saturated aliphatic hydrocarbon group containing a straight chain or branched chain of 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms (i.e., C 1-10 alkyl), further preferably 1 to 8 carbon atoms (i.e., C 1-8 alkyl), more preferably 1 to 6 carbon atoms (i.e., C 1-6 alkyl), more preferably 1 to 4 carbon atoms (i.e., C 1-4 alkyl), more preferably 1 to 3 carbon atoms (i.e., C 1-3 alkyl), for example "C 1-6alkyl" means that the group is an alkyl group and the number of carbon atoms in the carbon chain is between 1 and 6 (specifically 1, 2, 3, 4, 5, or 6). Examples include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-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, and the like.

[0155] "Halo-substituted alkyl" means "haloalkyl", such as C 1-6 alkyl substituted with halo is C 1-6 haloalkyl. "Haloalkyl" can be C 1-6 haloalkyl or C 1-4 haloalkyl.

[0156] The term "halogen" or "halo" means F, Cl, Br, I, unless otherwise specified. The term "haloalkyl" means an alkyl group as defined above in which one, two, or more hydrogen atoms are replaced by halogen. Representative examples of haloalkyl include CCl3, CF3, CHCl2, CH2Cl, CH2Br, CH2I, CH2CF3, CF2CF3, and the like.

[0157] The term "cycloalkyl" means, unless otherwise specified, a monocyclic saturated aliphatic radical having a specified number of carbon atoms, preferably containing 3 to 12 carbon atoms (i.e., C 3-12 cycloalkyl), more preferably 3 to 10 carbon atoms (C 3-10 cycloalkyl), further preferably 3 to 6 carbon atoms (C 3-6 cycloalkyl), 4 to 6 carbon atoms (C 4-6 cycloalkyl), and 5 to 6 carbon atoms (C 5-6 cycloalkyl). Examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, methylcyclopropyl, 2-ethyl-cyclopentyl, dimethylcyclobutyl, and the like.

[0158] The term "alkoxy" means, unless otherwise specified, an alkyl residue in which one or more carbons (and their associated hydrogens) are replaced by oxygen, such as "alkoxy", "alkoxyalkyl". Examples include methoxy, ethoxy, propoxy, methoxypropyl, and the like. "Alkoxy" can be C 1-6 alkoxy or C 1-4 alkoxy.

[0159] The term "alkylthio" means, unless otherwise specified, the replacement of the oxygen in the above "alkoxy" with sulfur or nitrogen. "Alkylthio" can be C 1-6 alkylthio or C 1-4 alkylthio.

[0160] The term "heterocyclyl," unless otherwise specified, refers to a saturated or partially unsaturated monocyclic, bicyclic, or polycyclic ringed cyclic hydrocarbon substituent, which is a non-aromatic structure, containing 3-20 ring atoms, of which 1, 2, 3, or more ring atoms are selected from N, O, or S, with the remaining ring atoms being C. Preferably, 3-12 ring atoms, further 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 are contained. Preferably, 1-4, more preferably 1-3 (i.e., 1, 2, or 3) heteroatoms are contained. Examples of monocyclic heterocyclyl groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, dihydropyrrolyl, piperidinyl, piperazinyl, pyranyl, and the like. Bicyclic or polycyclic heterocyclyl groups include spiro, fused, and bridged heterocyclyl groups.

[0161] The term "aryl," unless otherwise specified, refers to a monocyclic, bicyclic, and tricyclic aromatic carbocyclic ring 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, and the term "aryl" can be used interchangeably with the term "aromatic ring." Examples of aryl groups can include, but are not limited to, phenyl, naphthyl, anthryl, phenanthryl, or pyrenyl, and the like.

[0162] The term "heteroaryl," unless otherwise specified, refers to an aromatic monocyclic, bicyclic, or polycyclic ringed ring system containing a 5-16 membered structure, or a 5-14 membered structure, 5-12 membered structure, 5-10 membered structure, 5-8 membered structure, or a 5-6 membered structure, of which 1, 2, 3, or more ring atoms are heteroatoms and the remaining atoms are carbon, the heteroatoms being independently selected from O, N, or S, preferably 1, 2, or 3 heteroatoms are contained. Examples of heteroaryl groups can include, but are not limited to, furanyl, thiophenyl, oxazolyl, thiazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, thiadiazolyl, triazinyl, phtalazinyl, quinolinyl, isoquinolinyl, pteridinyl, purinyl, indolyl, isoindolyl, indazolyl, benzofuranyl, benzothiophenyl, benzopyridyl, benzopyrimidinyl, benzopyrazinyl, benzimidazolyl, benzophthalazinyl, pyrrolo[2,3-b]pyridyl, imidazo[l,2-a]pyridyl, pyrazolo[l,5-a]pyridyl, pyrazolo[l,5-a]pyrimidinyl, imidazo[l,2-b]pyridazinyl, [l,2,4]triazolo[4,3-b]pyridazinyl, [l,2,4]triazolo[l,5-a]pyrimidinyl, [l,2,4]triazolo[l,5-a]pyridyl, and the like.

[0163] The compounds described herein indicate that ring A contains at least NH2and -C(CH3)2OH substituents, and they are in adjacent positions on ring A. Other analogs.

[0164] 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.

[0165] 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.

[0166] 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.

[0167] The compounds of the application also include solvates or hydrates thereof, unless otherwise specified. The terms "solvate" or "solvate" refer to a physical association between one or more solvent molecules (either organic or inorganic) and a compound of the application. The physical association can be due to hydrogen bonding, to van der Waals forces, to dipole-dipole interactions, or to any other force capable of stabilizing the solvate. In certain instances, the solvate will be capable of being isolated, for example, when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. The solvent molecules can be present in a defined or an undefined arrangement. Solvates can include stoichiometric or non-stoichiometric amounts of the solvent molecules. "Solvate" embraces both solution-phase and isolatable solvates. Exemplary solvates include, but are not limited to, hydrates, ethanolates, methanolates, and isopropanolates. Methods for solvation are well known in the art.

[0168] The term "stereoisomer" refers to compounds having the same chemical constitution, but differing in the arrangement of atoms or groups in space, unless otherwise specified. Stereoisomers include enantiomers, diastereomers, conformers (rotamers), geometric isomers (cis / trans) isomers, atropisomers, and the like. Any mixture of the resulting stereoisomers can be separated, if desired, into the individual pure or substantially pure geometric isomers, enantiomers, diastereomers, by, for example, chromatatographic and / or fractional crystallization methods.

[0169] The term "tautomer" refers to structural isomers that exist in equilibrium with one another, capable of interconversion by a low energy barrier, unless otherwise specified. If tautomerism is possible (as in solution), a chemical equilibrium of the tautomers can be achieved. For example, prototropic tautomers (also known as proton-shift tautomers) include interconversions by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Bond tautomers include interconversions by reorganization of some bonding electrons.

[0170] Unless otherwise stated, the structural formulae described herein include all tautomeric forms (e.g., enantiomeric, diastereomeric, and geometric (or conformational) isomers): for example, the R, S configuration about asymmetric centers, the (Z), (E) isomers about double bonds, and the (Z), (E) conformational isomers. Accordingly, individual stereochemical isomers or mixtures thereof of the compounds of the application, or enantiomers, diastereomers, or geometric isomers (or conformational isomers) thereof, are within the scope of the application.

[0171] The compounds of the present application also include co-crystals thereof, and the term "co-crystal" is used to describe situations where neutral molecular components are present in the crystal in a definite stoichiometric ratio, unless otherwise specified. The preparation of pharmaceutical co-crystals enables changes to be made to the crystal form of an active pharmaceutical ingredient, which in turn can change its physicochemical properties without impairing its desired biological activity (see Pharmaceutical Salts and Co-crystals, J. Wouters and L. Quere, Eds., RSC Publishing, 2012).

[0172] The compounds of the present application also include polymorphs thereof, and the term "polymorph" refers to different arrangements of molecules of a chemical drug, generally manifested as different forms in which a drug substance exists in the solid state, unless otherwise specified. A drug can exist in more than one polymorphic state, and different polymorphic forms of the same drug can have different dissolution and absorption rates in the body, which can affect the dissolution and release of a formulation.

[0173] The compounds of the present application also include metabolites thereof, and the term "metabolite" refers to a product produced through metabolism of a specified compound or salt thereof in the body. Metabolic products of a compound can be identified using techniques well known in the art, and their activity can be characterized using assays as described herein. Such products can be oxidized, reduced, hydrolyzed, amidated, deamidated, esterified, deesterified, enzymatically cleaved, and the like. Accordingly, the present application includes metabolites of compounds, including those produced upon administration of the compound to a mammal for a period of time sufficient to yield a metabolic product.

[0174] The compounds of the present application also include prodrugs thereof, and the term "prodrug" refers to a drug that is converted into the parent drug in the body, unless otherwise specified. Prodrugs are often useful when the parent drug has poor solubility or stability, or when it is otherwise desirable to modify the physical properties or biological properties of the parent drug. Physical properties often relevant to solubility (either too high or too low lipid or water solubility) or stability, while problematic biological properties include too rapid metabolism or poor bioavailability, which can be related to physical chemical properties. For example, they can be bioavailable by oral administration whereas the parent is not. Prodrugs can also have improved solubility in pharmaceutical compositions compared to the parent drug. An example, without limitation, of a prodrug can be any of the compounds of the present application administered as an ester (a "prodrug") to facilitate transport across a cell membrane of a mammal, where water solubility is detrimental to mobility, but once inside the cell water solubility is beneficial for activity, and it is subsequently metabolically hydrolyzed to the carboxylic acid, the active entity. Another example of a prodrug can be a short peptide (polyamino acid) attached to an acid group, where the peptide is metabolized to reveal the active moiety.

[0175] Unless otherwise indicated, the term "optionally substituted" means that the hydrogen at the point of attachment of the group can be unsubstituted or substituted with one or more substituents preferably selected from the group consisting of halogen, hydroxy, thiol, cyano, nitro, amino, azido, oxo, carboxy, C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, C 3-10 cycloalkylsulfonyl, 3-10 membered heterocycloalkyl, C 6- 14 aryl or 5-10 membered heteroaromatic ring group, wherein the C 2-6 alkenyl, C 2-6 alkynyl, C 1-6 alkyl, C 1-6 alkoxy, C 3-10 cycloalkyl, C 3-10 cycloalkylsulfonyl, 3-10 membered heterocycloalkyl, C 6-14 aryl or 5-10 membered heteroaromatic ring group can be optionally substituted with one or more substituents selected from the group consisting of halogen, hydroxy, amino, cyano, C 1-6 alkyl or C 1-6 alkoxy, and the oxo means that two H at the same point of attachment are replaced by the same O to form a double bond.

[0176] The term "optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and thus the description includes instances where the event or circumstance occurs and instances where it does not.

[0177] The words "comprise" or "comprising" and variations such as "comprises" or "comprising", are to be interpreted, in open, non-exclusive sense, i.e. in the sense of "including, but not limited to".

[0178] The term "subject" can also be referred to as "individual" or "subject" refers to a cell or an animal, including but not limited to a mammal, such as a laboratory animal or a human. An "effective amount" refers to the amount of a compound or pharmaceutical composition described herein sufficient to achieve a desired application, including but not limited to the treatment of a disease or the alleviation of symptoms. In some embodiments, for example, the amount is a dose that can induce a particular response by a cell, or a dose that can exert a therapeutic effect on a disease in a model animal. This will vary depending on, for example, the particular compound chosen, the species and their age / existing health conditions of the subject, the dosing regimen followed, the severity of the disease, whether it is administered in combination with other agents, the time of administration, the tissue of administration, and the physical delivery system carrying it.

[0179] Some of the compounds in the present application have optical activity, and the compounds in the present application can be racemates, optical isomers or mixtures thereof. The synthesis of the optical isomers of the compounds in the present application can be prepared from starting materials of optical isomers or by separation of racemates.

[0180] The term "pharmaceutically acceptable carrier" also referred to as "pharmaceutically acceptable excipient" or "pharmaceutically acceptable vehicle" unless otherwise specified, refers to a generally-recognized medium in the art of delivering biologically-active agents to animals, particularly mammals. The pharmaceutically acceptable carrier is formulated according to a number of factors that are well within the purview of the ordinarily skilled artisan. These include, but are not limited to, the type and nature of the active agent being formulated, the subject to which the composition containing the agent is to be administered, the intended route of administration of the composition, and the therapeutic indication being targeted. Pharmaceutically acceptable carriers include aqueous and nonaqueous liquid media, as well as a variety of solid and semi-solid dosage forms. The preparation of the pharmaceutical compositions described herein, including, but not limited to, the compounds of the first aspect or the second aspect, or tautomers, stereoisomers, or pharmaceutically acceptable salts thereof, in admixture with a pharmaceutically acceptable carrier, is described herein.

[0181] The pharmaceutical compositions of the present application can be prepared by combining a compound of the present application with suitable pharmaceutically acceptable excipients.

[0182] Typical routes of administration of the compounds of the present application, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof, include, but are not limited to, oral, rectal, topical, inhalant, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0183] The pharmaceutical compositions of the present application can be manufactured in a manner that is itself known, e.g., by means of conventional mixing, dissolving, granulating, emulsifying, reching, lyophilizing processes, etc.

[0184] In all methods of administration of the compounds of the present application, the dose administered can range from 0.5 mg to 20 mg per person per day, in single or divided doses.

[0185] The above embodiments represent exemplary embodiments of the present application, but the present application is not limited to the above embodiments. In addition, each technical feature in the above embodiments of the present application can be combined with each other to constitute one or more new technical solutions, which also fall within the scope of the present application, as long as such new technical solutions are technically feasible. BRIEF DESCRIPTION OF DRAWINGS

[0186] Figure 1. The amount of tear secretion in a scopolamine-induced dry eye model test in mice

[0187] Figure 2 Scoring of corneal fluorescein staining in a mouse dry eye model induced by scopolamine DETAILED DESCRIPTION

[0188] The application will be further described with reference to the following examples. It should be appreciated that these examples are intended for illustration only and are not intended to limit the scope of the present application. Unless otherwise indicated, the experimental procedures in the following examples were carried out in accordance with conventional procedures or as recommended by the manufacturer. Unless otherwise defined, all other scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art. Further, any method and material similar or equivalent to those described herein can be used in the practice of the present application. The preferred methods and materials are described herein.

[0189] The structure of the compounds of the present application was determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS) or / and high performance liquid chromatography (HPLC). The NMR was measured by using a Bruker 400MHz or / and Varian 400MHz; the LC-MS was measured by using an Agilent, 1260 Infinity II-6120 / 6125 MSD; the HPLC was measured by using a Waters Acquity UPLC_2 or / and Shimadzu LC2030 or / and Agilent, 1260 Infinity II. The chiral compound resolution was measured by using an SFC-150 (Waters); the chiral column was DAICEL Chiralpak® OD; the column volume was 20 x 250 mm (10 pm particle size packing). The column volume was 20 x 250 mm (10 pm particle size packing).

[0190] The starting materials in the examples of the present application are known and commercially available or can be synthesized by or in accordance with methods known in the art.

[0191] The present application provides a method for preparing the compounds. The compounds can be prepared by the following steps.

[0192] Preparation Example 1

[0193] Preparation of 3-{[(tert-butoxy)carbonyl]amino}-4-(2-hydroxypropan-2-yl)benzoic acid (P-4):

[0194] First step: Compound P-1 (6.00 g) was dissolved in tetrahydrofuran (200 mL), methyl magnesium bromide in tetrahydrofuran (3 M, 52 mL) was added, the reaction was carried out at 0 °C for 0.5 h under nitrogen protection. After TLC detection, the reaction was quenched with water (100 mL), extracted with ethyl acetate, the organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, and then purified by flash chromatography (Silica gel, petroleum ether: ethyl acetate = 5:1) to give compound P-2 (5.50 g). LCMS (ESI) [M+H-H20] = 211.9. +

[0195] Second step: Compound P-2 (2.50 g) was dissolved in ethanol (50 mL), di-tert-butyl dicarbonate (3.41 g) was added, and the reaction was carried out at 40 °C for 16 h. The reaction was concentrated and then purified by flash chromatography (Silica gel, petroleum ether: ethyl acetate = 10:1) to give compound P-3 (3.00 g). LCMS (ESI) [M+H-t-Bu-H20] = 258.0; + 1 H NMR (400 MHz, DMSO-d6) δ 9.86 (s, 1H), 8.19 (d, J = 1.6 Hz, 1H), 7.19 - 7.09 (m, 2H), 6.17 (s, 1H), 1.49 (s, 6H), 1.47 (s, 9H).

[0196] Third step: Compound P-3 (3.00 g) was dissolved in dimethyl sulfoxide (30 mL), water (15 mL), triethylamine (6.38 mL), and dichloropalladium bis((cyclopentadien-1-yl) diphenylphosphine) iron (999.33 mg) was added, and the reaction was carried out at 80 °C for 16 h under carbon monoxide (balloon) atmosphere. The reaction was quenched with water (25 mL), extracted with ethyl acetate, the product was adjusted to pH = 5 with citric acid (3 M) in the aqueous phase, extracted with ethyl acetate (25 mL, 3 times), the organic phase was combined, dried over anhydrous sodium sulfate, filtered, concentrated, and then purified by flash chromatography (Silica gel, tetrahydrofuran: petroleum ether = 1:5) to give compound P-4 (584.99 mg). LCMS (ESI) [M+H-t-Bu-H20] = 222.1; + 1 H NMR (400 MHz, DMSO-d6) δ 12.88 (s, 1H), 9.80 (s, 1H), 8.58 (d, J = 1.5 Hz, 1H), 7.53 (dd, J = 8.2, 1.8 Hz, 1H), 7.35 (d, J = 8.2 Hz, 1H), 6.19 (s, 1H), 1.53 (s, 6H), 1.48 (s, 9H). ​​​

[0197] Preparation Example 2

[0198] Preparation of 4-{[(tert-butoxy)carbonyl]amino}-3-(2-hydroxypropan-2-yl)benzoic acid (P-8):

[0199] First step: Compound P-5 (6.00 g) was dissolved in tetrahydrofuran (200 mL), methyl magnesium bromide (43.47 mL) was added dropwise at 0 °C, and the reaction was carried out at 0 °C for 0.5 hours under nitrogen protection. After the reaction was completed, water (100 mL) was added to the reaction solution, and ethyl acetate was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and then separated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound P-6 (5.1 g). LCMS (ESI) [M+H-H20] = 212.1; + 1 H NMR (400 MHz, DMSO-d6) δ 7.11-7.01 (m, 2H), 6.57 (d, J = 8.4 Hz, 1H), 5.55 (s, 2H), 5.30 (s, 1H), 1.47 (s, 6H).

[0200] Second step: Compound P-6 (2.00 g) was dissolved in ethanol (20 mL) solvent, and di-tert-butyl dicarbonate (2.85 g) was added, and the reaction was carried out at 30 °C for 16 hours. After the reaction was completed, the reaction solution was directly concentrated, and separated and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain compound P-7 (2.30 g). LCMS (ESI) [M+H-t-Bu+Na] = 297.3; + 1 H NMR (400 MHz, DMSO-d6) δ 9.68 (s, 1H), 7.90 (d, J = 8.7 Hz, 1H), 7.42-7.36 (m, 1H), 7.34 (d, J = 2.3 Hz, 1H), 6.15 (s, 1H), 1.50 (s, 6H), 1.46 (s, 9H).

[0201] Third step: Compound P-7 (2.20 g) was dissolved in dimethyl sulfoxide (20 mL) and water (10 mL), and triethylamine (6.74 mL) was added, and palladium (2 + ​​)bis((cyclopenten-1,3-dien-1 -yl)diphenylphosphine)iron(0) (0.98 g), reaction under carbon monoxide (balloon pressure) atmosphere at 90 °C for 18 h. After the reaction was completed, water (60 mL) was added to dilute the reaction solution, and the pH was adjusted to 5 with a saturated aqueous citric acid solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether: tetrahydrofuran = 3:1) to obtain compound P-8 (581.51 mg). LCMS (ESI) [M+H-t-Bu-H2O] = 222.1. + 1 H NMR (400 MHz, DMSO-d6) δ 12.71 (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).

[0202] Example 1

[0203] Preparation of (2S)-2-{[4-({[3-amino-4-(2-hydroxypropan-2-yl)phenyl]formamido}methyl)- 2,6-dichlorophenyl]formamido}-3-(3-methanesulfonylphenyl)propanoic acid (Compound II-22):

[0204] First step: To a solution of compound P-4 (252.33 mg) in N,N-dimethylformamide was added HATU (2-(7-azabenzotriazol-1 -yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (406 mg), and the reaction was stirred for 0.166 h. Triethylamine (259.37 mg) and 10-1 (200 mg) were then added to the reaction solution, which was stirred at room temperature for 2 h. After the reaction was completed, the reaction solution was treated with water, and the resulting mixture was extracted with ethyl acetate. The combined organic phase was washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was separated by column chromatography on silica gel to obtain compound 10-2 (410 mg).

[0205] Second step: To a solution of compound 10-2 (410 mg) in dichloromethane (4 mL) was added trifluoroacetic acid (1 mL), and the reaction was carried out at 0 °C for 1.5 h. The pH was adjusted to 8 with an aqueous sodium bicarbonate solution, and the resulting mixture was extracted with ethyl acetate to obtain compound 10-3 (109 mg).

[0206] ​Step 3: To compound 10-3 (109 mg) was added methanol (3 mL) and water (1.5 mL). Lithium hydroxide 31.74 mg was added and the reaction was stirred at 70 °C for 2 hours. The reaction was diluted with water and the pH was adjusted to 4-5 with 1 M hydrochloric acid. The reaction was extracted with ethyl acetate and the organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure to give compound 10-4 (90 mg).

[0207] Step 4: To a solution of compound 10-4 (90 mg) in N,N-dimethylformamide was added HATU (2-(7-azabenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (107.68 mg) and the reaction was stirred for 0.166 hours. Triethylamine (68.78 mg) and (2S)-2-amino-3-(3-methanesulfonylphenyl)propionic acid benzyl ester (75.53 mg) were added to the reaction. The reaction was stirred at room temperature for 2 hours. After completion of the reaction, the reaction was treated with water and extracted with ethyl acetate. The organic layers were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by column chromatography on silica gel to give compound 10-5 (120 mg).

[0208] Step 5: To compound 10-5 (120 mg) was added methanol (3 mL) and water (1.5 mL). Lithium hydroxide (20.16 mg) was added and the reaction was stirred at 50 °C for 1 hour. The reaction was filtered and purified by preparative purification to give compound II-22 (50 mg). LCMS (ESI) [M+H] + = 622.0. 1 H NMR (400 MHz, DMSO-d6) δ 8.84 (t, J = 6.1 Hz, 1H), 8.60 (s, 1H), 7.83 (d, J = 1.8 Hz, 1H), 7.77 - 7.70 (m, 1H), 7.65 (d, J = 7.7 Hz, 1H), 7.53 (t, J = 7.7 Hz, 1H), 7.30 (s, 2H), 7.11 - 7.04 (m, 2H), 6.96 (dd, J = 8.1, 1.8 Hz, 1H), 5.59 (s, 2H), 5.31 (s, 1H), 4.57 (s, 1H), 4.38 (d, J = 5.9 Hz, 2H), 3.28 (dd, J = 13.8, 4.8 Hz, 1H), 3.13 (s, 3H), 3.06 (dd, J = 13.9, 8.7 Hz, 1H), 1.50 (s, 6H).

[0209] Example 2

[0210] Preparation of (2S)-2-{[4-({4-amino-3-(2-hydroxypropan-2-yl)phenyl]methylamino}methyl)-2,6-dichlorophenyl]methylamino}-3-(3-methanesulfonylphenyl)propanoic acid (Compound II-23):

[0211] Referring to example 1, compound P-4 was replaced by compound P-8 to give compound II-23 (32.72 mg). LCMS (ESI) [M+H] + = 622.8. 1 H NMR (400 MHz, DMSO-d6) δ 8.67 (t, J = 6.0 Hz, 1H), 8.53 (s, 1H), 7.82 (d, J = 1.8 Hz, 1H), 7.73 (dt, J = 7.8, 1.5 Hz, 1H), 7.67 - 7.61 (m, 1H), 7.58 (d, J = 2.1 Hz, 1H), 7.56 - 7.47 (m, 2H), 7.29 (s, 2H), 6.61 (d, J = 8.4 Hz, 1H), 5.95 (s, 2H), 5.30 (s, 1H), 4.55 (d, J = 6.4 Hz, 1H), 4.37 (d, J = 5.9 Hz, 2H), 3.26 (d, J = 4.8 Hz, 1H), 3.12 (s, 3H), 3.06 (dd, J = 13.9, 8.6 Hz, 2H), 1.51 (s, 6H).

[0212] Example 3

[0213] Preparation of (2S)-2-{[4-({[4-amino-2-fluoro-5-(2-hydroxypropan-2- yl)phenyl]formamido}methyl)-2,6-dichlorophenyl]formamido}-3-(3- methanesulfonylphenyl)propanoic acid (II-44):

[0214] First Step: Di-tert-butyl dicarbonate (9.11 mL) and triethylamine (4.22 mL), Raney nickel (306.14 mg) were added to a solution of compound 22-1 (2.4 g) in methanol (250 mL), which was stirred at room temperature for 1 hour under hydrogen (balloon pressure). The reaction solution was filtered, concentrated, and purified by flash chromatography (Silica gel, petroleum ether: ethyl acetate = 3: 1) to give compound 22-2 (3 g). LCMS (ESI) [M+H-Boc+ACN] + = 275.1

[0215] Second Step: Hydrogen chloride dioxane solution (15 mL, 4 M) was added to a solution of compound 22-2 (3.0 g) in dioxane (15 mL), which was stirred at room temperature for 1 hour. The reaction solution was concentrated, slurried with ethyl acetate (20 mL), filtered, and the filter residue was dried under vacuum to give compound 22-3 (1.8 g). LCMS (ESI) [M+H] + = 234.0.

[0216] Step 3: To a solution of compound 22-3 (900 mg) in acetonitrile (20 mL), 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (709.28 mg), TCFH (tetramethylchloro-urea hexafluorophosphate) (1400.1 mg) and 1-methylimidazole (0.82 mL) were added and the reaction mixture was stirred at 25 °C for 1 h. The reaction mixture was concentrated and purified by normal phase (petroleum ether: ethyl acetate = 1:1) to give compound 22-4 (830 mg). LCMS (ESI) [M+H] + = 429.1.

[0217] Step 4: To a solution of compound 22-4 (830 mg) in methanol (20 mL) and water (10 mL), lithium hydroxide (2315.38 mg) was added. The reaction mixture was heated to 100 °C and stirred for 1 h. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure, and the pH was adjusted to about 5 with 1 M hydrochloric acid solution, then extracted with ethyl acetate (15 mL*3), the combined organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 22-5 (750 mg). LCMS (ESI) [M+H] + = 415.1.

[0218] Step 5: To a solution of compound 22-7 (750 mg) in N,N-dimethylformamide (20 mL), 22-5 (1263.03 mg) and N,N-diisopropylethylamine (786.22 mg), HATU (2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (771.03 mg) were added, and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was directly purified by reverse phase (C18, acetonitrile / 0.1% formic acid aqueous system, 70% acetonitrile to give the product) to give compound 22-6 (1.0 g). LCMS (ESI) [M+H] + = 712.5

[0219] Step 6: To a solution of compound 22-6 (1 g) in tetrahydrofuran (10 mL) and water (5 mL), sodium hydroxide (164.4 mg) was added. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure, then the pH was adjusted to 5-6 with 1 M hydrochloric acid, and then purified by preparative separation (acetonitrile / 0.03% formic acid in water) to give compound II-44 (400 mg). LCMS (ESI) [M+H-H2O] + = 622.0; 1H NMR (400 MHz, DMSO-d6) δ 12.86 (s, 1H), 9.04 (d, J = 8.0 Hz, 1H), 8.37 - 8.24 (m, 1H), 7.86 (s, 1H), 7.76 (d, J = 7.7 Hz, 1H), 7.67 (d, J = 7.5 Hz, 1H), 7.56 (t, J = 7.7 Hz, 1H), 7.43 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 13.9 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.81 - 4.71 (m, 1H), 4.37 (d, J = 5.8 Hz, 2H), 3.29 - 3.25 (m, 1H), 3.14 (s, 3H), 3.01 (dd, J = 13.9, 10.5 Hz, 1H), 1.48 (s, 6H).

[0220] Example 4

[0221] Preparation of (S)-2-(4-((4-amino-2-fluoro-3-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-46):

[0222] Referring to the first step of Example 1, compound P-4 is replaced by compound 24-2 to obtain compound 24-3 (300 mg);

[0223] Second step: To the methanol solution (3 mL) of compound 24-3 (250 mg), a saturated solution of lithium hydroxide (3 mL) was added, the reaction was stirred at 100 °C for 12 hours, the reaction solution was cooled to room temperature, the pH was adjusted to weakly acidic with citric acid solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, the organic phase was removed under reduced pressure, the crude product was separated on a silica gel column to obtain compound 24-4 (120 mg);

[0224] Compound 24-4 (70 mg) was subjected to the fourth step and the fifth step of Example 1 to obtain compound II-46 (11.5 mg). LCMS (ESI) [M+H] = 642.4. + 1 ​H NMR (400 MHz, DMSO-d6) δ 8.70 (s, 1H), 8.32 (q, J = 6.0 Hz, 1H), 7.84 (d, J = 1.8 Hz, 1H), 7.74 (dt, J = 7.7, 1.5 Hz, 1H), 7.64 (s, 1H), 7.54 (s, 1H), 7.32 (s, 2H), 7.26 (s, 1H), 6.42 (d, J = 8.6 Hz, 3H), 5.68 (s, 1H), 4.61 (s, 1H), 4.35 (d, J = 6.0 Hz, 2H), 3.13 (s, 4H), 1.58 (d, J = 3.7 Hz, 6H).

[0225] Example 5

[0226] Preparation of (S)-2-(3-(2-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)ethyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-50):

[0227] First step: Allyl palladium chloride dimer (309.28 mg) was added to a solution of compound 28-1 (3 g), sodium 2-cyanoacetate (1.7 g), [5-(diphenylphosphoryl)-9,9-dimethyl-9H-xanthen-4-yl]diphenylphosphine (1.53 g) in mesitylene (20 mL) under nitrogen atmosphere at 120 °C for 5 h. After reaction, the mixture was filtered, the filtrate was added to water (100 mL), extracted with ethyl acetate (60 mL*3), the organic phase was combined, washed with saturated brine (100 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by reverse phase system (C18, 0.1% FA in water / MeCN = 40% / 60%) to give compound 28-2 (530 mg). 1 H NMR (400 MHz, CDCl3) δ 7.54 (d, J = 8.4 Hz, 1H), 7.41 (d, J = 8.4 Hz, 1H), 4.00 (s, 3H), 3.83 (s, 2H).

[0228] Second step: Raney nickel (1.86 g) was added to a solution of compound 28-2 (530 mg), di-tert-butyl dicarbonate (2.84 g), triethylamine (1.32 g) in methanol (10 mL) under hydrogen atmosphere at 25 °C for 0.5 h. The reaction mixture was filtered, the filtrate was concentrated, and the residue was purified by flash chromatography (Silica gel, PE / THF = 9 / 1) to give compound 28-3 (590 mg). LCMS (ESI) [M+H-t-Bu+ACN] = 332.8. + = 332.8.

[0229] Third step: Hydrochloric acid / 1.4-dioxane solution (6 mL, 4 M) was added to a solution of compound 28-3 (590 mg) in 1,4-dioxane (6 mL), and the mixture was reacted at 25 °C for 1 h. The reaction solution was filtered to collect the solid, and the filtrate was concentrated to give compound 28-4 (450 mg). LCMS (ESI) [M+H+ACN] = 289.1. +

[0230] Fourth step: N-Methylimidazole (127.07 mg) was added to a solution of 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (110 mg), compound 28-4 (146.82 mg), and TCFH (N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate) (217.14 mg) in acetonitrile (10 mL), and the mixture was reacted at 25 °C for 1 h. The reaction solution was concentrated, water (10 mL) was added, and the mixture was extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (silica gel, PE / THF = 2 / 1) to give compound 28-5 (270 mg). LCMS (ESI) [M+H] = 443.1. +

[0231] Fifth step: Potassium tert-butoxide (227.81 mg) was added to a solution of compound 28-5 (180 mg) in dimethyl sulfoxide (10 mL) and water (1 mL). The mixture was reacted at 60 °C for 2 h. The reaction solution was cooled to room temperature, adjusted to pH 3 with 1 M hydrochloric acid, extracted with ethyl acetate (10 mL*3), and the combined organic phase was washed with saturated brine (20 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 28-6 (100 mg). LCMS (ESI) [M+H] = 429.1. +

[0232] Sixth step: HATU (2-(7-Azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (106.29 mg) was added to a solution of compound 28-6 (80 mg), (S)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid hydrochloride (103.39 mg), and N,N-diisopropylethylamine (72.26 mg) in dimethylformamide (5 mL), and the mixture was reacted at 25 °C for 1 h. Water (20 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with saturated brine (20 mL*3). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by prep-TLC (PE / EA = 1 / 3) to give compound 28-7 (58 mg). LCMS (ESI) [M+H-H2O] = 577.2.​​​+ = 726.6.

[0233] Step 7: 1M aqueous sodium hydroxide solution (2 mL) was added to a solution of compound 28-7 (50 mg) in tetrahydrofuran (2 mL) at 25 °C for 1 h. The pH was adjusted to 7 with 1M hydrochloric acid, and the reaction was concentrated. The residue was purified by prep-HPLC (C18, 0.1% NH3.H2O in water / MeCN) to give compound II-50 (22.43 mg). LCMS (ESI) [M+H-H2O] + = 636.4. 1 H NMR (400 MHz, DMSO-d6) δ 7.78 (s, 1H), 7.75 - 7.66 (m, 2H), 7.61 (d, J = 7.5 Hz, 1H), 7.46 (t, J = 7.7 Hz, 2H), 7.39 - 7.33 (m, 2H), 7.26 (d, J = 8.1 Hz, 1H), 6.35 (d, J = 13.9 Hz, 1H), 6.12 (s, 2H), 5.39 (s, 1H), 4.11 (s, 1H), 3.46 - 3.40 (m, 2H), 3.27 - 3.22 (m, 2H), 3.11 (s, 3H), 2.90 (t, J = 6.9 Hz, 2H), 1.47 (s, 6H).

[0234] Example 6

[0235] Preparation of (S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)-N- methylbenzamide)methyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-54):

[0236] Step 1: Sodium hydride (179.53 mg) (60% content) was added to a solution of methyl 4-(((tert-butoxycarbonyl)amino)methyl)-2,6-dichlorobenzoate (37-1, 1 g) in dimethylformamide (20 mL) at 0 °C, and the temperature was maintained for 30 min. Iodomethane (637.07 mg) was slowly added dropwise to the system, and the reaction was carried out at 25 °C for 16 h. The reaction was quenched by adding water (30 mL), extracted with ethyl acetate (20 mL*3), washed with saturated brine (30 mL*3), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by flash chromatography (Silica gel, PE / THF = 10 / 1) to give compound 37-2 (560 mg). LCMS (ESI) [M+H-t-Bu+ACN] + = 333.1. 1H NMR (400 MHz, DMSO-d6) δ 7.39 (s, 2H), 4.40 (s, 2H), 3.92 (s, 3H), 2.82 (s, 3H), 1.46 - 1.34 (m, 9H).

[0237] Second step: hydrochloric acid / 1,4-dioxane (5 mL) was added to methyl 4-(((tert- butoxycarbonyl)(methyl)amino)methyl)-2,6-dichlorobenzoate (37-2, 590 mg) in 1,4- dioxane (5 mL) and reacted at 25 °C for 1 h. The reaction solution was concentrated to obtain compound 37-3 (450 mg). LCMS (ESI) [M+H] + = 248.0.

[0238] Third step: 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (37-4, 226 mg) was dissolved in acetonitrile (4 mL), and TCFH (N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate) (446.12 mg) and N-methylimidazole (261.07 mg) were added in turn, and finally 2,6-dichloro-4-[(methylamino)methyl]benzoic acid methyl ester hydrochloride (37-3, 315.59 mg) was added. The reaction was carried out at room temperature for 2 h. Water (10 mL) was added to quench the reaction, and the organic phase was extracted with ethyl acetate (5 mL*3), washed with saturated sodium chloride solution (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash chromatography (silica gel, petroleum ether: tetrahydrofuran = 3:1) to obtain compound 37-5 (230 mg). LCMS (ESI) [M+H-H2O] + = 425.2.

[0239] Fourth step: methyl 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)-N-methylbenzamido)methyl)- 2,6-dichlorobenzoate (37-5, 255 mg) was dissolved in methanol (3 mL), and lithium hydroxide (68.88 mg) and water (12 mL) were added in turn. The reaction solution was heated to 100 °C and reacted for 5 h. At 0 °C, the pH was adjusted to 3 with 1M hydrochloric acid, and the organic phase was extracted with ethyl acetate (10 mL*3), washed with saturated sodium chloride (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 37-6 (220 mg). LCMS (ESI) [M+H-H2O] + = 411.1.

[0240] Step 5: 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)-N-methylbenzamido)methyl)- 2,6-dichlorobenzoic acid (37-6, 220 mg) was dissolved in N,N dimethylformamide (1 mL), HATU (2-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (233.84 mg), triethylamine (155.58 mg) and (S)-2-amino-3-methylsulfanyl- benzenepropanoic acid benzyl ester hydrochloride (170.87 mg) were added successively. The reaction was stirred at room temperature for 2 hours. Water (10 mL) was added to quench the reaction, extracted with ethyl acetate (5 mL*3), washed with saturated sodium chloride (10 mL*3), dried over anhydrous sodium sulfate, filtered, concentrated. Compound 37-9 (122 mg) was obtained by column chromatography (silica gel, PE:EA = 1:2). LCMS (ESI) [M+H-H20] = 726.2. +

[0241] Step 6: (2S)-2-{[4-({l-[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]-N- methylformamido}methyl)-2,6-dichlorophenyl]formamido}-3-(3-methylsulfanylphenyl) propanoic acid benzyl ester (37-9, 122 mg) was dissolved in tetrahydrofuran (2 mL), sodium hydroxide (19.66 mg) and water (1 mL) were added. The reaction was stirred at room temperature for 2 hours. Concentrated, diluted with water (1 mL), at 0 °C, pH = 6 was adjusted with 1 M hydrochloric acid, purified by prep-HPLC (C-18, 0.1% NH4HCO3 in H2O / ACN) to give compound II-54 (27.17 mg). LCMS (ESI) [M+H-H20] = 636.1. + 1 H NMR (400 MHz, DMSO-d6) d 8.46 (s, 1H), 7.83 (s, 1H), 7.73 (d, J = 7.8 Hz, 1H), 7.65 (d, J = 7.7 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.24 (d, J = 49.7 Hz, 2H), 6.99 (s, 1H), 6.42 (d, J = 12.6 Hz, 1H), 6.02 (s, 2H), 5.36 (s, 1H), 4.52 (t, J = 38.4 Hz, 3H), 3.30 - 3.24 (m, 1H), 3.13 (s, 3H), 3.08 (dd, J = 13.8, 8.2 Hz, 1H), 2.82 (s, 3H), 1.47 (s, 6H).

[0242] Example 7

[0243] ​​Preparation of (S)-2-(4-((4-amino-N-ethyl-2-fluoro-5-(2-hydroxypropan-2- yl)benzamido)methyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-55):

[0244] First Step: To a solution of methyl 4-(((tert-butoxycarbonyl)amino)methyl)- 2,6-dichlorobenzoate (36-1, 1 g) in N,N-dimethylformamide (20 mL) was added sodium hydride (239.2 mg) under ice-bath, the reaction was stirred at this temperature for 0.5 h. Then iodomethane (932.7 mg) was added, the reaction was stirred at room temperature for 3.0 h. The reaction was diluted with water (50 mL), extracted with dichloromethane (50 mL*3), the organic phase was combined and washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried and concentrated, purified by flash chromatography (petroleum ether: tetrahydrofuran = 10:1) to give compound 36-2 (580 mg). LCMS (ESI) [M+H-t-Bu+ACN] = 347.0. + 1 H NMR (400 MHz, CDCl3) d 7.19 (s, 2H), 4.37 (s, 2H), 3.98 (d, J = 10.6 Hz, 3H), 3.24 (d, J = 32.3 Hz, 2H), 1.48 (s, 9H), 1.09 (t, J = 6.8 Hz, 3H).

[0245] Second Step: To a solution of methyl 4-(((tert-butoxycarbonyl)(ethyl)amino)methyl)- 2,6-dichlorobenzoate (36-2, 680 mg) in hydrochloric acid dioxane (2.0 M, 10 mL) was stirred at room temperature for 2.0 h. The reaction was concentrated to give compound 36-3 (420 mg). LCMS (ESI) [M+H] = 264.0. +

[0246] ​​Third step: To a solution of methyl 2,6-dichloro-4-(ethylamino)methyl)benzoate hydrochloride (36-3, 280.67 mg) in acetonitrile (20 mL) was added 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (200.4 mg), methylimidazole (154.35 mg,), TCFH (N,N,N',N'-tetramethyluronium hexafluorophosphate) (527.49 mg) at room temperature. The reaction was stirred at room temperature for 3.0 hours. To the reaction was added water (50 mL) to dilute, extracted with dichloromethane (30 mL*3), the combined organic phase was washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried, concentrated, purified by flash chromatography (petroleum ether: tetrahydrofuran = 10:1) to give compound 36-4 (190 mg). LCMS (ESI) [M+H-H2O] = 439.1. +

[0247] Fourth step: To a solution of methyl 4-((4-amino-N-ethyl-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzoate (36-4, 100 mg) in methanol (2 mL), water (8 mL) was added lithium hydroxide (26.19 mg), the temperature was raised to 100 °C and the reaction was stirred for 5 hours. To the reaction was added water (20 mL) to dilute, the reaction was adjusted to pH = 6 with 1M aqueous hydrochloric acid solution, extracted with ethyl acetate (20 mL*3), the combined organic phase was washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried, concentrated to give compound 36-5 (70 mg). LCMS (ESI) [M+H-H2O] = 425.2. +

[0248] Fifth step: To a solution of 4-((4-amino-N-ethyl-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzoic acid (36-5, 80 mg), (2S)-2-amino-3-(3-methylsulfonylphenyl)propanoic acid benzyl ester hydrochloride (66.75 mg) in N,N-dimethylformamide (10 mL) was added HATU (2-(7-azabenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (137.24 mg), triethylamine (36.52 mg) at room temperature. The reaction was stirred at 25 °C for 2 hours. To the reaction was added water (50 mL) to dilute, extracted with dichloromethane (10 mL*3), the combined organic phase was washed with saturated sodium chloride solution (10 mL*3), the organic phase was dried, concentrated, purified by flash chromatography (petroleum ether: tetrahydrofuran = 10:2) to give compound 36-7 (60 mg). LCMS (ESI) [M+H-H2O] = 740.1. +

[0249] ​​​Step 6: To a solution of ((S)-2-(4-((4-amino-N-ethyl-2-fluoro-5-(2-hydroxypropan-2- yl)benzamido)methyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (36-7, 60 mg) in tetrahydrofuran (8 mL), water (2 mL) was added sodium hydroxide (9.49 mg) at room temperature. The reaction was stirred at 25 °C for 1 h. The reaction was diluted with water (20 mL), and the pH was adjusted to 6 with 1 M aqueous hydrochloric acid solution. The reaction was extracted with ethyl acetate (30 mL*3), and the organic phase was washed with saturated sodium chloride solution (20 mL*3). The organic phase was dried and concentrated to give a crude product, which was purified by prep-HPLC (C18, 10 mmol / L FA in water, MeCN) to give compound II-55 (20 mg). LCMS (ESI) [M+H-H20] = 650.1. + 1 H NMR (400 MHz, DMSO-d6) d 12.89 (s, 1H), 9.06 (d, J = 7.5 Hz, 1H), 7.86 (s, 1H), 7.77 (d, J = 7.8 Hz, 1H), 7.67 (d, J = 7.8 Hz, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.31 (s, 2H), 6.94 (d, J = 7.9 Hz, 1H), 6.43 (d, J = 11.9 Hz, 1H), 5.98 (s, 2H), 5.35 (s, 1H), 4.76 (td, J = 10.0, 4.6 Hz, 1H), 4.53 (d, J = 70.7 Hz, 2H), 3.31 - 3.30 (m, 1H), 3.28 (d, J = 4.5 Hz, 1H), 3.16 (s, 1H), 3.14 (s, 3H), 3.01 (dt, J = 19.8, 9.9 Hz, 1H), 1.47 (s, 6H), 1.00 (s, 3H).

[0250] Example 8

[0251] Preparation of (S)-2-(4-((4-amino-N-cyclopropyl-2-fluoro-5-(2-hydroxypropan-2- yl)benzamido)methyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-56):

[0252] ​First Step: Dissolve 2,6-dichloro-4-formyl benzoic acid methyl ester (38-1, 2 g) in ethanol (60 mL), at 0 °C, add sodium borohydride (649.3 mg), after the addition, restore to room temperature, react for 2 hours. Concentrate the reaction solution, at 0 °C, slowly add saturated ammonium chloride (40 mL) to quench the reaction, extract with ethyl acetate (40 mL*3), combine the organic phase, wash with saturated sodium chloride (40 mL), dry over anhydrous sodium sulfate, filter, concentrate to obtain product 38-2 (2 g). LCMS (ESI) [M+H] + = 234.9.

[0253] Second Step: Dissolve 2,6-dichloro-4-hydroxymethyl benzoic acid methyl ester (38-2, 1.6 g) in acetonitrile (16 mL), then add dichlorosulfoxide (2.43 g). Heat the reaction solution to 70 °C and react for 2 hours. Add water (30 mL) to quench the reaction, extract with ethyl acetate (20 mL*3), combine the organic phase, wash with saturated sodium chloride (50 mL), dry over anhydrous sodium sulfate, filter, concentrate to obtain compound 38-3 (1.6 g).

[0254] Third Step: Dissolve 2,6-dichloro-4-(chloromethyl)benzoic acid methyl ester (38-3, 1.7 g) in acetonitrile (17 mL), then add cyclopropylamine (5 mL). Heat the reaction solution to 50 °C and react for 16 hours. Add water (30 mL) to quench the reaction, extract with ethyl acetate (15 mL*3), combine the organic phase, wash with saturated sodium chloride (30 mL), dry over anhydrous sodium sulfate, filter, concentrate. Purify by column chromatography (silica gel, PE:EA = 2:1) to obtain compound 38-4 (1.7 g). LCMS (ESI) [M+H] + = 274.0.

[0255] Fourth Step: Dissolve 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (38-5, 500 mg) in acetonitrile (5 mL), then add TCFH (N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate) (986.98 mg), N-methylimidazole (577.6 mg) and 2,6-dichloro-4-((cyclopropylamino)methyl)benzoic acid methyl ester (38-4, 642.89 mg) in sequence, stir at room temperature for 2 hours. Add water (20 mL) to quench the reaction, extract with ethyl acetate (15 mL*3), wash with saturated sodium chloride (30 mL), dry over anhydrous sodium sulfate, filter, concentrate. Purify by column chromatography (silica gel, PE:EA = 2:1) to obtain compound 38-6 (500 mg). LCMS (ESI) [M+H] + = 469.1.

[0256] Step 5: Methyl 4-((4-amino-N-cyclopropyl-2-fluoro-5-(2-hydroxypropan-2- yl)benzamido)methyl)-2,6-dichlorobenzoate (38-6, 200 mg) was dissolved in methanol (4 mL) and water (1 mL). Lithium hydroxide (51.49 mg) was added. The reaction was heated to 100 °C for 16 h. Concentration, water (2 mL) was added, at 0 °C, pH = 3 was adjusted with 1 M hydrochloric acid, a large amount of solid precipitated, the solid was filtered and dried to give compound 38-7 (100 mg, crude). LCMS (ESI) [M+H-H20] = 437.1. +

[0257] Step 6: 4-((4-amino-N-cyclopropyl-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzoic acid (38-7, 100 mg) was dissolved in N,N dimethylformamide (1 mL), HATU (2-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (125.27 mg), triethylamine (66.67 mg) and (S)-2-amino-3-methylsulfanyl- benzenepropanoic acid benzyl ester hydrochloride (73.22 mg) were added in sequence, stirred at room temperature for 2 h. Water (10 mL) was added to quench, extracted with ethyl acetate (15 mL*3), the organic phase was combined and washed with saturated sodium chloride (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated. Compound 38-9 (42 mg) was obtained by column chromatography (silica gel, PE:EA = 1:2). LCMS (ESI) [M+H-H20] = 752.2. +

[0258] Step 7: (S)-2-(4-((4-amino-N-cyclopropyl-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (38-9, 45 mg) was dissolved in tetrahydrofuran (1 mL) and water (0.2 mL), sodium hydroxide (12 mg) was added, stirred at room temperature for 2 h. Concentration, water (1 mL) was added, pH = 6 was adjusted with 1 M hydrochloric acid, tetrahydrofuran (0.5 mL) was added and purified by prep-HPLC (C-18, 0.1% NH4HCO3 in H2O / ACN) to give compound II-56 (8.37 mg). LCMS (ESI) [M+H-H20] = 662.2; + 1 ​​​H NMR (400 MHz, DMSO-d6) δ 12.83 (s, 1H), 9.03 (s, 1H), 7.87 (s, 1H), 7.77 (d, J = 7.8 Hz, 1H), 7.68 (d, J = 7.7 Hz, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.29 (s, 2H), 6.99 (d, J = 8.0 Hz, 1H), 6.40 (d, J = 12.7 Hz, 1H), 6.01 (s, 2H), 5.35 (s, 1H), 4.76 (s, 1H), 4.61 (s, 2H), 3.29 - 3.28 (m, 1H), 3.14 (s, 3H), 3.03 (dd, J = 13.9, 10.3 Hz, 1H), 2.64 (s, 1H), 1.48 (s, 6H), 0.53 - 0.41 (m, 4H).

[0259] Example 9

[0260] Preparation of ((S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl- d2)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-59):

[0261] First step: 2,6-dichloro-4-(cyanomethyl)benzoic acid methyl ester (44-1, 2.0 g) was dissolved in tetrahydrofuran (20.0 mL), then cooled to -20 °C, and lithium aluminum deuteride (442.34 mg) was slowly added. The reaction was stirred at -20 °C for 1 hour. The reaction solution was added to saturated ammonium chloride solution (30 mL), and then dichloromethane (30 mL) was added for extraction. The organic phase was rotary evaporated to obtain the crude product. The crude product was separated by reverse phase preparation to obtain compound 44-2 (510 mg). LCMS (ESI) [M+H] + = 236.04

[0262] Second step: 4-aminomethyl-D2-2,6-dichlorobenzoic acid methyl ester (44-2, 510 mg) was added to a 25 mL single-neck flask, acetonitrile (5.0 mL) was added, and then 4-amino-2-fluoro-5-(2-hydroxy-2-propyl)benzoic acid (44-3, 460.57 mg) was added. Then TCFH (N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate) (909.16 mg) and azidomethylimidazole (532 mg) were added, and the reaction was continued at 25 °C for 16.0 hours. The reaction solution was added to 20 mL of water, and then 30 mL of ethyl acetate was added for extraction. The organic phase was dried and evaporated to dryness. The product was separated by column (petroleum ether-THF = 1:2) to obtain compound 44-4 (320 mg). LCMS (ESI) [M+H] + = 431.14.

[0263] Step 3: 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl-d2)- 2,6-dichlorobenzoic acid methyl ester (44-4, 320.0 mg) was dissolved in water (10.0 mL), methanol (10.0 mL) was added, then lithium hydroxide monohydrate (177.7 mg) was added, and the reaction was continued at 100 °C for 2 hours. The reaction solution was adjusted to pH about 8 using 1M dilute hydrochloric acid solution, then extracted with ethyl acetate (20.0 mL) three times, and the organic phase was dried with anhydrous sodium sulfate. After the reaction solution was rotary evaporated, the crude compound was obtained. Compound 44-5 (190 mg) was obtained by column separation using a petroleum ether-tetrahydrofuran system (petroleum ether: tetrahydrofuran = 1:3). LCMS (ESI) [M+H] + = 417.17

[0264] Step 4: 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl-d2)- 2,6-dichlorobenzoic acid (44-5, 190 mg) was dissolved in N,N-dimethylformamide (6.0 mL), then (S)-benzyl 2-amino-3-(3-(methylsulfonyl)phenyl)propanoate hydrochloride (151.8 mg) was added, HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (259.7 mg) was added, and finally N,N-diisopropylethylamine (176.6 mg) was added. The reaction was continued at 25 °C for 4.0 hours. The reaction solution was added with water 10 mL, then extracted with 20 mL of ethyl acetate, and the organic phase was evaporated to dryness. Compound 44-7 (120.0 mg) was obtained by column separation using a petroleum ether-tetrahydrofuran system (1:3). LCMS (ESI) [M+H] + = 714.25

[0265] Step 5: (S)-benzyl 2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl-d2)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoate (44-7, 120.0 mg) was dissolved in water (5.0 mL), ethanol (5.0 mL) was added, then lithium hydroxide monohydrate (11.5 mg) was added, and the reaction was continued at 25 °C for 1 hour. The reaction solution was adjusted to pH about 7 using 1M dilute hydrochloric acid solution, then extracted with ethyl acetate (20.0 mL) three times, and the organic phase was dried with anhydrous sodium sulfate. After the reaction solution was rotary evaporated, the crude compound was obtained. Compound II-59 (60 mg) was obtained by column separation and purification (C18, 0.1% NH4HCO3 in water / CH3CN). LCMS (ESI) [M+H] + = 624.15; 1H NMR (400 MHz, DMSO-d6) δ 8.77 (s, 1H), 8.27 (t, J = 6.3 Hz, 1H), 7.84 (t, J = 1.8 Hz, 1H), 7.74 (dt, J = 7.8, 1.5 Hz, 1H), 7.65 (dt, J = 7.8, 1.4 Hz, 1H), 7.54 (t, J = 7.7 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 13.9 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.67 - 4.62 (m, 1H), 3.30 - 3.26 (m, 1H), 3.13 (s, 2H), 3.07 - 3.01 (m, 1H), 1.48 (s, 6H).

[0266] Example 10

[0267] Preparation of (2S)-2-{[4-({[3-amino-4-(2-hydroxypropan-2-yl)phenyl]methyl}amino- carbonyl)-2,6-dichlorophenyl]formamido}-3-(3-methanesulfonylphenyl)propanoic acid (II-67):

[0268] First step: 2,6-dichlorobenzene-1,4-dicarboxylic acid dimethyl ester (30-1, 1000 mg) was dissolved in tetrahydrofuran (10 mL) and water (10 mL) at room temperature, sodium hydroxide (152.1 mg) was added, stirred at room temperature for 6 hours. The reaction solution was concentrated, 1M hydrochloric acid was added to adjust the pH to 5, extracted with ethyl acetate (20 mL) for three times, the organic phase was combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated to obtain the crude compound 30-2 (900 mg). 1 H NMR (400 MHz, DMSO-d6) δ 7.91 (s, 2H), 3.93 (s, 3H).

[0269] Second step: tetra(triphenylphosphine) palladium (100.44 mg) was added to a solution of zinc cyanide (153.08 mg) and 2-(2-amino-4-bromophenyl)propan-2-ol (30-3, 200 mg) in N,N-dimethylformamide (5 mL), stirred at 130°C for 18 hours under nitrogen protection. The reaction solution was filtered, washed with ethyl acetate, the filtrate was diluted with water, extracted with ethyl acetate (10 mL*3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and then purified by flash chromatography (Silica gel, petroleum ether: tetrahydrofuran = 10:1) to obtain the target compound, which was concentrated to obtain compound 30-4 (50 mg). LCMS (ESI) [M+H-H2O] = 159.2 + = 159.2

[0270] Step 3: Ammonia (0.5 mL) and Raney nickel (166.52 mg) were added to a solution of 3-amino-4-(2-hydroxypropan-2-yl)benzonitrile (30-4, 500 mg) in methanol (30 mL) and stirred at room temperature under hydrogen (balloon pressure) for 18 h. Filtered, washed with methanol, and concentrated the filtrate to give crude compound 30-5 (200 mg). LCMS (ESI) [M+H-H20] + = 163.3; 1 H NMR (400 MHz, DMSO-d6) δ 6.91 (d, J = 7.9 Hz, 1H), 6.54 (s, 1H), 6.43 (d, J = 7.8 Hz, 1H), 5.32 (d, J = 7.8 Hz, 2H), 5.12 (s, 1H), 3.53 (s, 2H), 1.47 (s, 6H).

[0271] Step 4: 3,5-Dichloro-4-(methoxycarbonyl)benzoic acid (250 mg) was dissolved in N,N dimethylformamide (5 mL) and 2-[2-amino-4-(aminomethyl)phenyl]propan-2-ol (30-5, 180.94 mg), HATU (2-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (572.52 mg), N,N-diisopropylethylamine (389.2 mg) were added and stirred at room temperature for 3 h. The reaction was quenched with water (10 mL) and extracted with ethyl acetate (20 mL) three times. The organic phase was combined, washed with saturated aqueous sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash chromatography (Silica gel, tetrahydrofuran: petroleum ether = 1:3) to give compound 30-6 (250 mg). LCMS (ESI) [M+H] + = 393.1.

[0272] Step 5: Methyl 4-({[3-amino-4-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)-2,6- dichlorobenzoate (30-6, 250 mg) was dissolved in dimethyl sulfoxide (5 mL) and water (0.5 mL) and potassium tert-butoxide (204.62 mg) was added and stirred at 60 °C for 1 h. The reaction was adjusted to pH 6 with 1 M aqueous hydrochloric acid and purified directly by reverse phase C18 column (acetonitrile / 0.05% aqueous TFA system, 43% for product) to give compound 30-7 (120 mg). LCMS (ESI) [M+H-H20] + = 379.2

[0273] Step 6: 4-({[3-amino-4-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)-2,6- dichlorobenzoic acid (30-7, 115 mg) was dissolved in N,N dimethylformamide (5 mL) at room temperature, HATU (2-(7-azabenzotriazol-1-yl)-1,1,3,3- tetramethyluronium hexafluorophosphate) (165.11 mg), benzyl 2-amino-3-(3- methylsulfonylphenyl)propanoate hydrochloride (107.26 mg), N,N-diisopropylethylamine (DIEA, 0.16 mL,) were added, stirred at room temperature for 3 hours. The reaction was quenched with water (20 mL) and extracted with ethyl acetate (20 mL) for three times. The organic phase was combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by flash chromatography (Silica gel, tetrahydrofuran: petroleum ether = 1:1) to give compound 30-8 (150 mg). LCMS (ESI) [M+H-H20] + = 694.2.

[0274] Step 7: (2S)-2-{[4-({[3-amino-4-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)- 2,6-dichlorophenyl]formamido}-3-(3-methylsulfonylphenyl)propanoic acid benzyl ester (30-8, 150 mg) was dissolved in tetrahydrofuran (12.5 mL) at room temperature, aqueous sodium hydroxide solution (1 M, 2.5 mL) was added, stirred at room temperature for 3 hours. The reaction was adjusted to pH 7 with 1 M aqueous hydrochloric acid solution and purified by prep-HPLC (C18, 10 mmol / L NH4HCO3 in water, MeCN) to give compound II-67 (83.82 mg). LCMS (ESI) [M+H] + = 622.3; 1 H NMR (400 MHz, DMSO-d6) δ 9.14 (t, J = 5.8 Hz, 1H), 8.73 (br. s, 1H), 7.88 (s, 2H), 7.84 (s, 1H), 7.74 (d, J = 7.8 Hz, 1H), 7.66 (d, J = 7.8 Hz, 1H), 7.54 (t, J = 7.7 Hz, 1H), 6.94 (d, J = 7.9 Hz, 1H), 6.52 (d, J = 1.3 Hz, 1H), 6.42 (dd, J = 7.9, 1.4 Hz, 1H), 5.19 (br. s, 2H), 4.66 - 4.55 (m, 1H), 4.29 (d, J = 5.7 Hz, 2H), 3.32 - 3.29 (m, 2H), 3.14 (s, 3H), 3.11 - 3.05 (m, 1H), 1.46 (s, 6H).

[0275] Example 11

[0276] Preparation of (S)-2-(4-((4-amino-3-(2-hydroxypropan-2-yl)benzyl)carbamoyl)-2,6- dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-68):

[0277] First step: To a solution of 2-(2-amino-5-bromophenyl)propan-2-ol (31-1, 2 g) in N,N-dimethylformamide (20 mL), zinc cyanide (1.02 g) and tetrakis(triphenylphosphine)palladium (2.01 g) were added, and stirred at 130 °C for 18 hours under nitrogen protection. The reaction system was filtered with celite, water (10 mL) was added to the reaction solution, extracted with ethyl acetate (10 mL*3), the combined organic phase was washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried, concentrated, and purified by normal phase (petroleum ether: tetrahydrofuran = 20%) to obtain compound 31-2 (1.4 g). LCMS (ESI) [M+H-H2O] + = 159.6. 1 HNMR (400MHz, DMSO-d6) δ 7.32 (d, J = 8.7 Hz, 2H), 6.68 (d, J = 8.1 Hz, 1H), 6.35 (s, 2H), 5.44 (s, 1H), 1.48 (s, 6H).

[0278] Second step: To a solution of 3-amino-4-(2-hydroxypropan-2-yl)benzonitrile (31-2, 500 mg) in methanol (6 mL), Raney nickel (16.65 mg), ammonia water (0.3 mL) and triethylamine (861.34 mg) were added, and stirred at room temperature for 18 hours under hydrogen protection. The reaction system was filtered and concentrated to obtain compound 31-3 (480 mg). LCMS (ESI) [M+H-H2O] + = 164.2. 1 H NMR (400MHz, DMSO-d6) δ 6.91 (s, 1H), 6.82 (m, 1H), 6.48 (d, J = 8.0 Hz, 1H), 5.21 (d, J = 13.9 Hz, 2H), 5.10 (s, 1H), 3.47 (t, J = 7.2 Hz, 2H), 1.45 (s, 6H).

[0279] Step 3: To a solution of 3,5-dichloro-4-(methoxycarbonyl)benzoic acid (31-3, 345.42 mg) in N,N-dimethylformamide (6 mL) was added ethyl bis(prop-2-yl)amine (537.75 mg) and HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (1.05 g), after stirring for 10 minutes, 2-[2-amino-5-(aminomethyl)phenyl]propan-2-ol (250 mg) was added, the reaction was stirred at room temperature for 50 minutes. To the reaction was added water (10 mL), extracted with ethyl acetate (10 mL*3), the combined organic phase was washed with saturated sodium chloride solution (10 mL*3), the organic phase was dried, concentrated, purified by normal phase separation (petroleum ether: tetrahydrofuran = 40%), to give compound 31-5 (250 mg). LCMS (ESI) [M+H-H2O] + = 393.3; 1 H NMR (400 MHz, DMSO-d6) δ 9.15 (s, 1H), 8.02 (s, 2H), 7.01 (d, J = 1.6 Hz, 1H), 6.93 - 6.88 (m, 1H), 6.56 (d, J = 8.1 Hz, 1H), 5.36 (s, 2H), 5.18 (s, 1H), 4.30 (d, J = 5.7 Hz, 2H), 3.95 (s, 3H), 1.49 (s, 6H).

[0280] Step 4: To a solution of methyl 4-({[4-amino-3-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)- 2,6-dichlorobenzoate (31-5, 200 mg) in dimethyl sulfoxide: water = (5:1) (3 mL) was added potassium tert-butoxide (163.7 mg), which was stirred at 60 °C for 1 hour. Purified by reverse phase separation (acetonitrile: water (trifluoroacetic acid) = 40%), to give compound 31-6 (150 mg). LCMS (ESI) [M+H-H2O] + = 379.1.

[0281] Step 5: To a solution of 4-({[4-amino-3-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)- 2,6-dichlorobenzoic acid (31-6, 200 mg) in N,N-dimethylformamide (5 mL) was added benzyl (2S)-2-amino-3-(3-methanesulfonylphenyl)propanoate (167.85 mg), after 10 min, ethyl bis(propan-2-yl)amine (195.2 mg) and HATU (2-(7-azabenzotriazol-1- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (287.15 mg) were added, the reaction was stirred at 25 °C for 50 min. To the reaction was added water (5 mL), extracted with ethyl acetate (5 mL*3), the organic phase was combined, washed with saturated sodium chloride solution (5 mL*3), the organic phase was dried, concentrated, purified by TLC (petroleum ether: tetrahydrofuran = 1:2) to give compound 31-8 (150 mg). LCMS (ESI) [M+H-H2O] = 694.2. + 1 H NMR (400 MHz, DMSO-d6) δ 9.34 - 9.29 (m, 1H), 9.15 - 9.04 (m, 1H), 7.96 - 7.95 (m, 3H), 7.89 - 7.84 (m, 3H), 7.78 (d, J = 7.8 Hz, 1H), 7.67 (d, J = 7.1 Hz, 1H), 7.56 (t, J = 7.7 Hz, 1H), 7.39 - 7.35 (m, 5H), 6.99 - 6.92 (m, 1H), 6.88 (d, J = 7.2 Hz, 1H), 6.57 - 6.51 (m, 1H), 5.18 (s, 2H), 4.96 (d, J = 2.8 Hz, 1H), 4.28 (t, J = 6.3 Hz, 2H), 3.16 - 3.04 (m, 5H), 1.47 (d, J = 3.8 Hz, 6H).

[0282] Step 6: To (2S)-2-{[4-({[4-amino-3-(2-hydroxypropan-2-yl)phenyl]methyl}carbamoyl)- 2,6-dichlorophenyl]formamido}-3-(3-methanesulfonylphenyl)propanoic acid benzyl ester (31-8, 150 mg) was added aqueous sodium hydroxide (1 M): tetrahydrofuran = (1:1) (4 mL), which was stirred at 25 °C for 2 h, the reaction was adjusted to neutral with 1 M hydrochloric acid solution. Purified by Prep-HPLC (0.1% NH4HCO3 in water, ACN) to give compound II-68 (12.4 mg).

[0283] Compound II-68: LCMS (ESI) [M+H-H2O] = 604.4. + 1 ​​H NMR (400 MHz, DMSO-d6) δ 9.06 (s, 1H), 7.85 (s, 2H), 7.80 (s, 1H), 7.70 (m, 1H), 7.62 (m, 1H), 7.55 - 7.47 (m, 1H), 6.99 (s, 1H), 6.87 (m, 1H), 6.55 (d, J = 7.9 Hz, 1H), 5.32 (s, 2H), 5.17 (s, 1H), 4.27 (d, J = 5.5 Hz, 2H), 3.29 - 3.25 (m, 2H), 3.16 - 3.12 (m, 4H), 1.48 (s, 6H).

[0284] Example 12

[0285] Preparation of (2S)-2-(4-(((4-amino-3-(2-hydroxypropan-2-yl)phenyl)(methyl)phosphoryl)ethynyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-70):

[0286] First Step: (4-amino-3-(2-hydroxypropan-2-yl)phenyl)(methyl)phosphonic acid ethyl ester (33-1, 2 g) was added to DMF (5 mL), replaced with nitrogen for three times, sodium hydride (1.34 g) was added under ice bath, stirred for 30 min, 1-(chloromethyl)-4-methoxybenzene (6 g) was added under ice bath, continued to stir for 1.5 h. The reaction solution was added to methanol for quenching, separated and purified by Prep-HPLC (C18, 10 mmol / L NH4HCO3 in water, MeCN) to give compound 33-2 (0.72 g). LCMS (ESI) [M+H] = 378.3. +

[0287] Second Step: (4-amino-3-(2-((4-methoxybenzyl)oxy)propan-2-yl)phenyl)(methyl)phosphonic acid ethyl ester (33-2, 270 mg) was added to DCM (1 mL), replaced with nitrogen for three times, thionyl chloride (2 mL) was added under ice bath, stirred at room temperature for 16 h. The reaction solution was concentrated under reduced pressure to give compound 33-3 (270 mg, crude). Used directly for next step.

[0288] ​Step 3: Methyl 2,6-dichloro-4-ethynylbenzoate (500 mg) was added to THF (5 mL), replaced with nitrogen for three times, n-butyllithium solution (0.83 mL, 2.5 M) was added dropwise at -78 °C, stirring was continued for 20 min, (4-amino-3-(2-((4-methoxybenzyl)oxy)prop-2-yl)phenyl)(methyl)phosphinic chloride (33-3, 380 mg) in THF (4 mL) was added, stirring was continued for 20 min. The reaction was quenched by saturated aqueous sodium bicarbonate solution, extracted with DCM, concentrated under reduced pressure to give the crude product. The crude product was purified by silica gel column (DCM:MeOH = 5:1) to give compound 33-5 (50 mg). LCMS (ESI) [M+H] + = 440.2.

[0289] Step 4: Methyl 4-(((4-amino-3-(2-hydroxyprop-2-yl)phenyl)(methyl)phosphinic acetyl)-2,6-dichlorobenzoate (33-5, 45 mg), lithium iodide (50 mg) were added to pyridine (2 mL), stirring at 120 °C for 3 h. The reaction was concentrated under reduced pressure to give compound 33-6 (45 mg, crude). Used directly for next step.

[0290] Step 5: 4-(((4-amino-3-(2-hydroxyprop-2-yl)phenyl)(methyl)phosphinic acetyl)-2,6-dichlorobenzoic acid (33-6, 45 mg) was added to DMF (1 mL), (S)-benzyl 2-amino-3-(3-(methylsulfonyl)phenyl)propanoate (70 mg), HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (100 mg) and DIPEA (0.19 mL) were added, stirring at room temperature for 4 h. The reaction was purified by Prep-HPLC (C18, 10 mmol / L NH4HCO3 in water, MeCN) to give compound 33-8 (10 mg). LCMS (ESI) [M+H] + = 741.3 & 743.3.

[0291] Sixth step: (2S)-2-(4-(((4-amino-3-(2-hydroxypropan-2-yl)phenyl)(methyl)phosphoryl)ethynyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (33-8, 10 mg) was added to DCM (2 mL), nitrogen was purged for three times, boron tribromide (0.13 mL) was added slowly at -78 °C, stirring was continued for 0.5 h. The reaction was quenched by methanol at -78 °C, the pH was adjusted to >8 with saturated aqueous sodium carbonate solution, the solution was separated and purified by prep-HPLC (C18, 10 mmol / L NH4HCO3 in water, MeCN) to give compound II-70 (5.3 mg). LCMS (ESI) [M+H] + = 651.2; 1 H NMR (400 MHz, DMSO-d6) δ 8.09 (s, 1H), 7.78 (d, J = 1.9 Hz, 1H), 7.73 - 7.66 (m, 2H), 7.61 (d, J = 7.6 Hz, 1H), 7.52 - 7.36 (m, 3H), 7.20 (s, 1H), 6.75 (dd, J = 8.2, 3.4 Hz, 1H), 6.11 (s, 2H), 5.38 (s, 1H), 4.28 (d, J = 5.5 Hz, 1H), 3.25 - 3.10 (m, 5H), 1.89 (d, J = 14.4 Hz, 3H), 1.38 (d, J = 111.1 Hz, 6H).

[0292] Examples 13, 14

[0293] Preparation of (2S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(S-methylsulfonimidoyl)phenyl)propanoic acid (II-71) and (2S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6- dichlorobenzamido)-3-(3-(R-methylsulfonimidoyl)phenyl)propanoic acid (II-72):

[0294] First step: (2S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(S-methylsulfonimidoyl)phenyl)propanoic acid (II-88, 250 mg) was resolved by SFC (15 40% MeOH DEA C6 IC) to give compound II-71 (t = 3.391 min, 67.2 mg) and II-72 (t = 4.537 min, 36.12 mg).

[0295] Instrument system: Waters SFC 150; Column name: Chiralpak® IC; Column dimensions: 250*30 mm 10 pm; Mobile phase A: Supercritical carbon dioxide; Mobile phase B: MEOH (+0.1% DEA); Mobile phase ratio 60:40; Detection wavelength: 214 nm; Flow rate: 140 ml / min; Column temperature: room temperature; Back pressure: 100 bar; Injection volume: 1 mL; Cycle time: 8 min; Solvents: Methanol (reagent grade), Supercritical carbon dioxide (food grade); Sample preparation: Sample dissolved in 13 mL methanol.

[0296] II-71 (or II-72): LCMS (ESI) [M+H] + = 639.2; 1 H NMR (400 MHz, DMSO-d6) δ 12.87 (s, 1H), 9.02 (d, J = 7.9 Hz, 1H), 8.32 - 8.25 (m, 1H), 7.87 (s, 1H), 7.80 - 7.74 (m, 1H), 7.61 - 7.56 (m, 1H), 7.51 (t, J = 7.7 Hz, 1H), 7.43 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 14.0 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.78 - 4.66 (m, 1H), 4.37 (d, J = 5.9 Hz, 2H), 4.10 (s, 1H), 3.28 - 3.23 (m, 1H), 3.04 - 2.97 (m, 4H), 1.48 (s, 6H).

[0297] II-72 (or II-71): LCMS (ESI) [M+H] + = 639.2; 1 H NMR (400 MHz, DMSO-d6) δ 12.86 (s, 1H), 8.97 (s, 1H), 8.33 - 8.23 (m, 1H), 7.87 (s, 1H), 7.79 - 7.74 (m, 1H), 7.60 - 7.55 (m, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.43 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 14.0 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.73 - 4.64 (m, 1H), 4.37 (d, J = 5.9 Hz, 2H), 4.09 (s, 1H), 3.27 - 3.22 (m, 1H), 3.03 - 2.97 (m, 4H), 1.48 (s, 6H).

[0298] Example 15​

[0299] Preparation of (S)-2-(4-((4-amino-3-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-73):

[0300] First step: Dissolve the starting material 4-((tert-butoxycarbonyl)amino)-3-fluoro-5-(2- hydroxypropan-2-yl)benzoic acid (59-1, 3 g) in acetonitrile (60 mL), add 4-(aminomethyl)- 2,6-dichlorobenzoic acid methyl ester hydrochloride (3.11 g), TCFH (N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate) (4.03 g), N-methylimidazole (2.36 g) in sequence, stir the reaction system at room temperature for 1 hour, concentrate to remove acetonitrile, add water (50 mL), extract with dichloromethane (60 mL*3), dry the combined organic phase with anhydrous sodium sulfate, filter and concentrate to obtain the crude product, which is separated and purified by silica gel column (petroleum ether: tetrahydrofuran = 1:1) to obtain compound 59-2 (3.1 g). LCMS (ESI) [M+H-t-Bu-H2O+ACN] = 496.1 +

[0301] Second step: Dissolve the starting material 4-((4-((tert-butoxycarbonyl)amino)-3-fluoro-5-(2- hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzoic acid methyl ester (59-2, 3.1 g) in methanol (100 mL) and water (50 mL), add lithium hydroxide (7012.32 mg), stir the reaction system at 100°C for 48 hours, filter, adjust the pH of the filtrate to 5 with 1M aqueous citric acid solution, extract with ethyl acetate (90 mL*3), dry the combined organic phase with anhydrous sodium sulfate, filter and concentrate to obtain the crude product, which is separated and purified by reverse phase system (0.1% FA in water / ACN) to obtain compound 59-3 (450 mg). LCMS (ESI) [M+H] +

[0302] ​​Step 3: Dissolve the starting material 4-((4-amino-3-fluoro-5-(2-hydroxypropan-2- yl)benzamido)methyl)-2,6-dichlorobenzoic acid (59-3, 500 mg) in N,N- dimethylformamide (10 mL), add 2-amino-3-(3-methanesulfonylphenyl)propionic acid benzyl ester hydrochloride (534.43 mg), HATU (2-(7-azabenzotriazol-1- yl)-1,1,3,3-hexafluoro-phosphonic 6-oxide hexahydrate) (686.77 mg), N,N- diisopropylethylamine (466.86 mg) in sequence, stir the reaction mixture at room temperature for 1 hour, purify the reaction mixture directly by reverse phase system (0.1% FA in water / ACN) to give compound 59-4 (400 mg). LCMS (ESI) [M+H-H20] = 712.1. +

[0303] Step 4: Dissolve the starting material (S)-2-(4-((4-amino-3-fluoro-5-(2- hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl) phenyl)propionic acid benzyl ester (450 mg) in tetrahydrofuran (59-4, 4.5 mL) and 1 M sodium hydroxide (4.5 mL), stir the reaction mixture at room temperature for 1 hour, concentrate to remove tetrahydrofuran, adjust pH to neutral with 1 M hydrochloric acid, concentrate to give a crude product, purify the crude product by prep-HPLC (10 mmol NH4HCO3 in water / ACN) to give compound II-73 (181.42 mg). LCMS (ESI) [M+H-H20] = 621.9. + 1 HNMR (400 MHz, DMSO-d6) δ 8.83 (s, 1H), 8.52 (s, 1H), 7.83 (s, 1H), 7.73 (d, J = 7.8 Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.56 - 7.46 (m, 3H), 7.30 (s, 2H), 5.88 (s, 2H), 5.52 (s, 1H), 4.56 (s, 1H), 4.38 (d, J = 5.8 Hz, 2H), 3.31 - 3.25 (m, 1H), 3.14 - 3.04 (m, 4H), 1.53 (s, 6H).

[0304] Example 16

[0305] Preparation of (S)-2-(4-((4-amino-2,3-difluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-74):

[0306] ​​Step 1: To a solution of methyl 4-(aminomethyl)-2,6-dichlorobenzoate hydrochloride (40-1, 200 mg) in acetonitrile (5 mL), 4-amino-2,3-difluoro-5-(2-hydroxypropyl-2-yl)benzoic acid (170.92 mg), 1-methyl-1H-imidazolium (182.08 mg), and TCFH (N,N,N',N'-tetramethylchlorourea hexafluorophosphate) (311.13 mg) were added sequentially. The reaction mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated and purified by normal-phase silica gel column chromatography (dichloromethane:methanol = 30:1) to give compound 40-2 (290 mg). LCMS (ESI) [M+H] + =447.1; 1 H NMR (400MHz, DMSO-d) 6 )δ8.55(s,1H),7.50(s,2H),7.25(d,J=7.6Hz,1H),6.14(s,2H),5.56(s,1H),4.45(d,J=6.0Hz,2H),3.91(s,3H),1.50(s,6H).

[0307] Step 2: An aqueous solution of lithium hydroxide (1313.42 mg) was added to a methanol (2 mL) solution of methyl 4-({[4-amino-2,3-difluoro-5-(2-hydroxypropyl-2-yl)phenyl]formamido}methyl)-2,6-dichlorobenzoate (40-2, 280 mg). The reaction mixture was heated under reflux at 100 °C for 2 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure, and the pH was adjusted to approximately 5 with 1 M hydrochloric acid. The solution was then extracted with ethyl acetate (5 mL x 3). The combined organic phases were washed with saturated brine (5 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound 40-3 (220 mg). LCMS (ESI) [MH] - =431.1; 1 H NMR (400MHz, DMSO-d6) δ14.11(s,1H),8.54(s,1H),7.42(s,2H),7.25(d,J=7.2Hz,1H),6.13(s,2H),4.46–4.40(m,2H),1.50(s,6H).

[0308] Step 3: To a solution of 4-({[4-amino-2,3-difluoro-5-(2-hydroxypropan-2- yl)phenyl]formamido}methyl)-2,6-dichlorobenzoic acid (40-3, 220.95 mg) in N,N- dimethylformamide (5 mL) was added triethylamine (206.43 mg), HATU (2-(7-azabenzotriazol- yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (387.83 mg) and (2S)-2-amino-3-(3- methylsulfonylphenyl)propionic acid benzyl ester hydrochloride (282.94 mg) and the reaction was stirred at room temperature for 2 hours. The reaction was washed with saturated sodium chloride (5 mL) three times and extracted with ethyl acetate (5 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel plate chromatography to give compound 40-5 (240 mg). LCMS (ESI) [M-H] - = 748.5.

[0309] Step 4: To a solution of (2S)-2-{[4-({[4-amino-2,3-difluoro-5-(2-hydroxypropan-2- yl)phenyl]formamido}methyl)-2,6-dichlorophenyl]methylamino}-3-(3-methylsulfonylphenyl)propionic acid benzyl ester (40-5, 150 mg) in tetrahydrofuran (2 mL) was added a solution of sodium hydroxide (24.04 mg) in water (1 mL) and the reaction was stirred at room temperature for 2 hours. The reaction was concentrated under reduced pressure, adjusted to pH 5-6 with 1 M hydrochloric acid and purified by preparative separation (acetonitrile / 0.03% formic acid in water) to give compound II-74 (105 mg). LCMS (ESI) [M-H] - = 656.1; 1 H NMR (400 MHz, DMSO-d6) δ 12.90 (s, 1H), 9.00 (s, 1H), 8.53 (d, J = 5.6 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.56 (t, J = 7.6 Hz, 1H), 7.34 (s, 2H), 7.23 (d, J = 7.2 Hz, 1H), 6.13 (s, 1H), 5.56 (s, 0.7H), 4.74 (s, 1H), 4.42 - 4.38 (m, 2H), 3.90 (s, 1H), 3.28 - 3.26 (m, 1H), 3.14 (s, 3H), 3.07 - 2.97 (m, 1H), 1.49 (s, 6H).

[0310] Example 17

[0311] Preparation of (2S)-2-[4-(1-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)-N-methylbenzamide)ethyl)-2,6-dichlorobenzamide]-3-(3-(methanesulfonyl)phenyl)propionic acid (II-75):

[0312] Step 1: Under ice bath conditions, methyl 2,6-dichloro-4-carboxybenzoate (41-1, 2 g) was dissolved in tetrahydrofuran (20 mL). After purging the reaction solution with nitrogen, methyl Grignard reagent (3 M in Me-THF, 2.86 mL) was added dropwise. The reaction solution was brought to room temperature and stirred for 2 hours. The reaction solution was quenched with saturated ammonium chloride (8 mL) solution. The resulting white suspension was filtered through diatomaceous earth, and the filtrate was diluted with water (50 mL). Then, it was extracted with ethyl acetate (50 mL * 3). The organic phases were combined and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated. The residue was purified by column chromatography (ethyl acetate / petroleum ether = 0–14%) to give compound 41-2 (1.83 g). LCMS (ESI) [M+H] + =248.8.

[0313] Step 2: Methyl 2,6-dichloro-4-(1-hydroxyethyl)benzoate (41-2, 1.4 g) was dissolved in dichloromethane (20 mL), and Desmond reagent (3.58 g) was added to the reaction solution. The mixture was stirred at room temperature for 2 hours. The reaction solution was quenched with saturated sodium bicarbonate (10 mL), and the resulting white suspension was filtered through diatomaceous earth. The filtrate was diluted with water (50 mL), and then extracted with dichloromethane (50 mL * 3). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (ethyl acetate / petroleum ether = 0–20%) to give compound 41-3 (1.2 g). LCMS (ESI) [M+H] + =247.0.

[0314] Step 3: Methyl 4-acetyl-2,6-dichlorobenzoate (41-3, 1.2 g) was dissolved in methanol (20 mL), and methylamine hydrochloride (2.9 g) was added to the reaction solution. The mixture was stirred at room temperature for 16 hours. Sodium cyanoborohydride (0.84 g) was added to the reaction solution. The mixture was stirred at 90 °C for 6 hours. The reaction solution was quenched with saturated sodium bicarbonate (100 mL) solution, then extracted with ethyl acetate (50 mL * 3), the organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated, and the residue was purified by column chromatography (ethyl acetate / petroleum ether = 0–100%) to give compound 41-4 (0.69 g). LCMS (ESI) [M+H] + =263.8.

[0315] Fourth Step: To a solution of methyl 2,6-dichloro-4-(1-(methylamino)ethyl)benzoate (41-4, 690 mg) in acetonitrile (10 mL), 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (561.23 mg), TCFH (N,N,N',N'-tetramethylchloroformamidinium hexafluorophosphate) (1.1 g) and 1-methylimidazole (648.33 mg) were added, the reaction was stirred at 25 °C for 2 hours. Concentration, purification by normal phase (petroleum ether: ethyl acetate = 5:3) gave compound 41-5 (590 mg). LCMS (ESI) [M+H-H20] + = 439.0.

[0316] Fifth Step: To a mixture solution of methyl 4-(1-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)-n-methylbenzoyl)ethyl)-2,6-dichlorobenzoate (41-5, 590 mg) in methanol (20 mL) and water (10 mL), lithium hydroxide (2706.68 mg) was added. The reaction mixture was heated to 100 °C and stirred for 1 hour. After the reaction mixture was cooled to room temperature, it was concentrated under reduced pressure, then the pH was adjusted to about 5 with 1M hydrochloric acid solution, and then extracted with ethyl acetate (15 mL*3), the organic phase was combined and washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 41-6 (530 mg). LCMS (ESI) [M+H-H20] + = 424.9.

[0317] Sixth Step: To a solution of (2S)-benzyl 2-amino-3-(3-methylsulfonylphenyl)propanoate hydrochloride (600.72 mg) in N,N-dimethylformamide (10 mL), 4-(1-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)-n-methylbenzylamino)ethyl)-2,6-dichlorobenzoic acid (41-6, 480 mg) and ethyl bis(prop-2-yl)amine (1.11 g) were added, and HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (617.56 mg), the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was directly purified by reverse phase (C18, acetonitrile / 0.1% formic acid aqueous system, 70% acetonitrile to give the product). Compound 41-8 (330 mg) was obtained. LCMS (ESI) [M+H] + = 760.1.

[0318] Seventh step: To a solution of benzyl (2S)-2-(4-(l-(4-amino-2-fluoro-5-(2- hydroxypropan-2-yl)-n-methylbenzamide)ethyl)-2,6-dichlorobenzamide)-3-(3- (methylsulfonyl)phenyl)propanoate (41-8, 0.33 g) in tetrahydrofuran (6 mL) and water (3 mL) was added sodium hydroxide (51.6 mg). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, then adjusted to pH 5-6 with 1 M hydrochloric acid, and purified by preparative separation (acetonitrile / 0.03% aqueous formic acid) to give compound II-75 (9.78 mg). LCMS (ESI) [M+H-H20] + = 650.0; 1 H NMR (400 MHz, DMSO-d6) δ 8.73 - 8.60 (m, 1H), 7.84 (s, 1H), 7.75 (d, J = 8.0 Hz, 1H), 7.66 (d, J = 8.0 Hz, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.35 - 7.20 (m, 2H), 6.97 (d, J = 8.0 Hz, 1H), 6.43 (d, J = 12.0 Hz, 1H), 6.01 (s, 2H), 5.35 (s, 1H), 4.64 - 4.56 (m, 1H), 3.28 - 3.24 (m, 2H), 3.13 (s, 3H), 3.10 - 3.04 (m, 2H), 2.62 (s, 3H), 1.50 (d, J = 8.0 Hz, 3H), 1.47 (s, 6H).

[0319] Example 18

[0320] Preparation of (2S)-2-{[4-({l-[4-amino-2,3-difluoro-5-(2-hydroxypropan-2- yl)phenyl]-N-methylcarboxamido}methyl)-2,6-dichlorophenyl]carboxamido}-3-(3- methylsulfonylphenyl)propanoic acid (II-76):

[0321] First step: Di-tert-butyl dicarbonate (4 mL) and triethylamine (2.44 mL), nickel (127.65 mg) were added to a solution of methyl 2,6-dichloro-4-cyanobenzoate (42-1, 1 g) in methanol (200 mL, 100.0%) and it was stirred at 25 °C for 2 hours under hydrogen (balloon pressure). The reaction mixture was filtered, concentrated, and purified by flash chromatography (silica gel, petroleum ether: ethyl acetate = 3: 1) to give compound 42-2 (1000 mg). LCMS (ESI) [M+H-t-Bu+ACN] + = 318.8.

[0322] Second step: Sodium hydride (85.99 mg) was added to a solution of methyl 2,6-dichloro-4-[[(2-methylpropan-2-yl)oxycarbonylamino]methyl]benzoate (42-2, 500 mg) in N,N-dimethylformamide (2 mL, 100.0%) at 0 °C, and stirred for 0.5 h. Then iodomethane (424.72 mg) was added to the reaction solution, and the mixture was stirred at 25 °C for 16 h. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution (2 mL) at 0 °C, extracted with ethyl acetate (3 mL*3), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was directly purified by reverse phase separation (C18, acetonitrile / 0.1% formic acid aqueous system, 80% acetonitrile content to produce the product) to give compound 42-3 (300 mg). LCMS (ESI) [M+H-t-Bu+ACN] = 333.0. +

[0323] Third step: To a solution of methyl 4-({[(tert-butoxy)carbonyl](methyl)amino}methyl)-2,6-dichlorobenzoate (42-3, 200 mg) in dioxane (3 mL, 100.0%) was added hydrogen chloride dioxane solution (1.4 mL, 4 M) at room temperature, and the mixture was stirred at 25 °C for 2 h. The reaction was concentrated, slurried with ethyl acetate (1 mL), filtered, and the filter residue was dried under vacuum to give compound 42-4 (130 mg). LCMS (ESI) [M+H] +

[0324] Fourth step: To a solution of methyl 2,6-dichloro-4-[(methylamino)methyl]benzoate hydrochloride (42-4, 80 mg) in acetonitrile (2 mL) was added 4-amino-2,3-difluoro-5-(2-hydroxypropan-2-yl)benzoic acid (65 mg), TCFH (N,N,N',N'-tetramethylchloro urium hexafluorophosphate) (118.32 mg), and 1-methylimidazole (69.24 mg), and the mixture was stirred at 25 °C for 2 h. The reaction was concentrated and purified by normal phase separation (petroleum ether: ethyl acetate = 1:1) to give compound 42-5 (100 mg). LCMS (ESI) [M+H] +

[0325] ​​​Fifth Step: To a mixture of methyl 4-({l-[4-amino-2,3-difluoro-5-(2- hydroxypropan-2-yl)phenyl]-N-methylcarbamimidoyl}methyl)-2,6-dichlorobenzoate (42-5, 250 mg) in methanol (2 mL) and water (2 mL) was added lithium hydroxide (129.8 mg). The reaction mixture was heated to 100 °C and stirred for 1 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and then adjusted to pH 5 with 1 M hydrochloric acid solution. The mixture was extracted with ethyl acetate (15 mL*3), and the combined organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 42-6 (230 mg). LCMS (ESI) [M+H] + = 446.9, [M+H-H20] + = 428.9.

[0326] Sixth Step: To a solution of (2S)-2-amino-3-(3-methylsulfonylphenyl)propanoic acid benzyl ester hydrochloride (235.68 mg) in N,N-dimethylformamide (2 mL) was added 4-({l-[4-amino-2,3-difluoro-5-(2-hydroxypropan-2-yl)phenyl]-N-methylcarbamimidoyl}methyl)-2,6-dichlorobenzoic acid (42-6, 190 mg) and triethylamine (171.95 mg), HATU (2-(7-azabenzotriazolyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (323.05 mg), and the reaction mixture was stirred at room temperature for 1 h. The reaction mixture was directly purified by reverse phase separation (C18, acetonitrile / 0.1% formic acid aqueous system, 70% acetonitrile to elute the product) to give compound 42-8 (70 mg). LCMS (ESI) [M+H-H20] + = 744.1.

[0327] Seventh Step: To a solution of (2S)-2-{[4-({l-[4-amino-2,3-difluoro-5-(2- hydroxypropan-2-yl)phenyl]-N-methylcarbamimidoyl}methyl)-2,6-dichlorophenyl]formamido}-3-(3-methylsulfonylphenyl)propanoic acid benzyl ester (42-8, 60 mg) in tetrahydrofuran (1 mL) and water (1 mL) was added sodium hydroxide (9.44 mg). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure, and then adjusted to pH 5-6 with 1 M hydrochloric acid. The resulting mixture was purified by preparative separation (acetonitrile / 0.03% formic acid in water) to give compound II-76 (7.5 mg). LCMS (ESI) [M+H-H20] + = 654.0; 1H NMR (400MHz, DMSO-d6) δ12.90(s,1H),9.05(s,1H),7.87(s,1H),7.77(d,J=7. 6Hz,1H),7.67(d,J=7.6Hz,1H),7.59-7.55(m,1H),7.33(s,1H),7.19-7.17(m, 2H),6.86(s,1H),5.97(s,2H),5.55(s,1H),4.78-4.76(m,1H),4.75-4.63(m,2 H), 3.28 (d, J = 4.4Hz, 1H), 3.14 (s, 3H), 3.03 (m, 1H), 2.84 (s, 3H), 1.49 (s, 6H).

[0328] Example 19

[0329] (2S)-2-[({4-[({[4-amino-2-fluoro-5-(prop-1-en-2-yl)phenyl]carbonyl}amino)methyl]-2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyldioxane-λ] 6 Preparation of [-thio)phenyl]propionic acid (II-77):

[0330] Step 1: Dissolve (2S)-2-[({4-[({[4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)phenyl]carbonyl}amino)methyl]-2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyldioxane-λ6-thio)phenyl]propionic acid (II-44, 200 mg) in 1,4-dioxane (5.0 mL), add water (5.0 mL), then add HCl (0.24 mL) (4.0 M 1,4-dioxane solution), and continue the reaction at 100 °C for 16 hours. Adjust the pH of the reaction solution to approximately 8 with 1 M sodium hydroxide solution, filter, and then use preparative separation and purification (C18, 0.1% NH4HCO3 in water / CH3CN) to obtain compound II-77 (20.0 mg). LCMS (ESI) [M+H] + =622.16; 1H NMR (400 MHz, DMSO-d6) δ 9.06 (d, J = 8.3 Hz, 1H), 8.36 - 8.28 (m, 1H), 7.86 (t, J = 1.9 Hz, 1H), 7.76 (dt, J = 7.7, 1.5 Hz, 1H), 7.69 - 7.64 (m, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.33 - 7.29 (m, 3H), 6.43 (d, J = 14.0 Hz, 1H), 5.25 - 5.23 (m, 1H), 4.97 (d, J = 2.2 Hz, 1H), 4.80 - 4.74 (m, 1H), 4.37 (d, J = 6.0 Hz, 2H), 3.33 - 3.27 (m, 1H), 3.14 (s, 3H), 3.04 - 2.98 (m, 1H), 1.98 (s, 3H).

[0331] Example 20

[0332] Preparation of (2S)-2-({[4-({[(4-amino-2-fluorophenyl)carbonyl]amino}methyl)-2,6- dichlorophenyl]carbonyl}amino)-3-[3-(methyldioxido-λ6-sulfanyl)phenyl]propanoic acid (II-78):

[0333] First step: (2S)-2-[({4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]-2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyldioxido-λ6-sulfanyl)phenyl]propanoic acid (II-44, 200 mg) was dissolved in 1,4-dioxane (5.0 mL), water (5.0 mL) was added, then HCl (0.24 mL) (4.0 M in 1,4-dioxane) was added, and the reaction was continued at 100 °C for 16 hours. The reaction solution was adjusted to pH about 8 with 1 M sodium hydroxide solution, and after filtration, preparative separation was performed (C18, 0.1% NH4HCO3 in water / CH3CN) to obtain compound II-78 (50.0 mg). LCMS (ESI) [M+H] + = 582.12; 1H NMR (400 MHz, DMSO-d6) δ 9.07 (d, J = 8.3 Hz, 1H), 8.31 - 8.27 (m, 1H), 7.86 (t, J = 2.0 Hz, 1H), 7.77 (dt, J = 7.8, 1.5 Hz, 1H), 7.67 (dt, J = 7.7, 1.5 Hz, 1H), 7.57 (t, J = 7.7 Hz, 1H), 7.44 (t, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (dd, J = 8.5, 2.1 Hz, 1H), 6.31 (dd, J = 14.3, 2.1 Hz, 1H), 4.79 - 4.75 (m, 1H), 4.37 (d, J = 6.0 Hz, 2H), 3.29 (dd, J = 14.1, 4.7 Hz, 1H), 3.17 (s, 3H), 3.04 - 2.96 (m, 1H).

[0334] Example 21

[0335] (2R)-2-[({4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]- 2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyldioxo-λ 6 Preparation of (2R)-2-[({4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]- 2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyldioxo-λ

[0336] First step: 4-(aminomethyl)-2,6-dichlorobenzoic acid methyl ester (46-1, 500.0 mg) was dissolved in acetonitrile (5.0 mL), then 4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzoic acid (455.42 mg), TCFH (N,N,N',N'-tetramethylchloroformamidium hexafluorophosphate) (898.99 mg) and azidomethyl imidazole (526.1 mg) were added, and the reaction was stirred at 25 °C for 16 hours. The reaction solution was added with water (20 mL), then dichloromethane (20.0 mL) was added for extraction, and the organic phase was rotary evaporated to obtain a crude product, which was separated by column using a petroleum ether-ethyl acetate (petroleum ether: ethyl acetate = 1:1) system to obtain compound 46-3 (560 mg). LCMS (ESI) [M+H] + = 429.15.

[0337] Second step: 4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]- 2,6-dichlorobenzoic acid methyl ester (46-3, 1000.0 mg) was dissolved in water (10.0 mL), methanol (10.0 mL) was added, then lithium hydroxide monohydrate (111.58 mg) was added, and the reaction was continued at 100 °C for 16 hours. The reaction solution was adjusted to pH = 8 with 1M dilute hydrochloric acid solution, and then extracted with ethyl acetate (20.0 mL) three times. The organic phase was dried with anhydrous sodium sulfate, and the reaction solution was rotary dried to obtain the crude compound. The compound 46-4 (420 mg) was separated by column chromatography using a petroleum ether-tetrahydrofuran system (petroleum ether: tetrahydrofuran = 1:3). LCMS (ESI) [M+H] + = 415.16.

[0338] Third step: 4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]- 2,6-dichlorobenzoic acid (46-4, 320 mg) was dissolved in N,N-dimethylformamide (10.0 mL), then (2R)-2-amino-3-[3-(methyl-dioxo-λ6-sulfanyl)phenyl]propanoic acid benzyl ester (256.93 mg) was added, HATU (2-(7-azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (439.53 mg) was added, and finally N,N-diisopropylethylamine (298.79 mg) was added. The reaction was continued at 25 °C for 16.0 hours. The reaction solution was added with 10 mL of water, and then extracted with 20 mL of ethyl acetate. The organic phase was evaporated to dryness, and the compound (46-6, 330.0 mg) was obtained by column chromatography using dichloromethane-methanol (20:1). LCMS (ESI) [M+H] + = 730.21.

[0339] Fourth step: (2R)-2-[({4-[({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]carbonyl}amino)methyl]- 2,6-dichlorophenyl}carbonyl)amino]-3-[3-(methyl-dioxo-λ6-sulfanyl)phenyl]propanoic acid benzyl ester (46-6, 330.0 mg) was dissolved in anhydrous ethanol (5.0 mL), water (5.0 mL) was added, and then lithium hydroxide monohydrate (56.86 mg) was added. The reaction was continued at 25 °C for 2.0 hours. The reaction solution was adjusted to pH = 8 with 1M dilute hydrochloric acid, and then filtered. Compound II-79 (220.0 mg) was obtained by preparative separation (C18, 0.1% NH4HCO3 in water / CH3CN). LCMS (ESI) [M+H-H2O] + = 622.17; 1H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.30-8.25 (m, 1H), 7.83 (t, J = 1.9 Hz, 1H), 7.73 (dt, J = 7.8, 1.5 Hz, 1H), 7.64 (dt, J = 7.7, 1.4 Hz, 1H), 7.52 (t, J = 7.7 Hz, 1H), 7.44 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 13.9 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.56-4.51 (m, 1H), 4.37 (d, J = 6.0 Hz, 2H), 3.30-3.25 (m, 1H), 3.12 (s, 3H), 3.09-3.04 (m, 1H), 1.48 (s, 6H).

[0340] Example 22

[0341] Preparation of (S)-2-(4-(4-amino-2-fluoro-5-(3-hydroxypentan-3-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-80):

[0342] First Step: Methyl 2-amino-5-bromo-4-fluorobenzoate (47-1, 500 mg) was dissolved in tetrahydrofuran (10 mL) under ice bath, the reaction solution was purged by nitrogen and ethyl formate (3.4 M in THF, 3.56 mL) was added dropwise. The reaction solution was allowed to warm to room temperature and stirred for 4 hours. The reaction solution was quenched by saturated ammonium chloride solution (10 mL), the resulting white suspension was filtered by celite, the filtrate was diluted with water (50 mL) and extracted with ethyl acetate (10 mL*3), the organic phase was combined and dried over anhydrous sodium sulfate, filtered and concentrated, the resulting residue was purified by column chromatography (ethyl acetate / petroleum ether = 0~15%) to give compound 47-2 (530 mg). LCMS (ESI) [M+H-H2O] = 259.8. +

[0343] Second Step: Di-tert-butyl dicarbonate (1.26 g) was added to 3-(2-amino-5-bromo-4- fluorophenyl)pentan-3-ol (47-2, 530 mg) in ethanol (10 mL) and reacted at 60 °C for 16 hours. The reaction solution was allowed to cool to room temperature and concentrated under reduced pressure, the resulting residue was purified by reverse phase (acetonitrile / 0.1% trifluoroacetic acid in water) to give compound 47-3 (650 mg). LCMS (ESI) [M+H-t-Bu-H2O] = 301.9. +

[0344] ​​Third step: To a mixture of tert-butyl (4-bromo-5-fluoro-2-(3-hydroxypentan-3- yl)phenyl)carbamate (47-3, 650 mg) in dimethyl sulfoxide (10 mL) and water (5 mL) was added triethylamine (1.75 g) and dichloro palladium (2+) bis ((cyclopentadien-1- yl) diphenylphosphine) iron (253 mg) respectively. The reaction mixture was stirred at 100 °C for 16 hours under the protection of carbon monoxide gas. When the reaction mixture was cooled to room temperature, saturated sodium bicarbonate solution was added to the reaction mixture to adjust the pH to about 9, then water (20 mL) was added to dilute the solution, and ethyl acetate (25 mL*3) was used to extract some impurities. The aqueous phase was adjusted to pH about 4 with 1 M hydrochloric acid solution, and then was continuously extracted with ethyl acetate (25 mL*3). The organic phase was washed with water (40 mL*2) and saturated brine (40 mL*2) respectively, dried over anhydrous sodium sulfate, filtered and concentrated. The obtained residue was purified by column chromatography (ethyl acetate / petroleum ether = 1:1) and reverse phase (acetonitrile / 0.03% formic acid in water = 50%) respectively to give compound 47-4 (220 mg). LCMS (ESI) [M-H] - = 340.0.

[0345] Fourth step: To a solution of 4-(tert-butoxycarbonyl)amino-2-fluoro-5-(3- hydroxypentan-3-yl)benzoic acid (47-4, 170 mg) in acetonitrile (4 mL) was added methyl 4-(aminomethyl)-2,6-dichlorobenzoate hydrochloride (134.7 mg), TCFH (N,N,N',N'-tetramethylchloro urium hexafluorophosphate) (209.6 mg) and 1-methylimidazole (122.7 mg), and the reaction was carried out at 25 °C for 2 hours. Concentration and purification by normal phase separation (petroleum ether: ethyl acetate = 2:1) gave compound 47-6 (230 mg). LCMS (ESI) [M+H] + = 557.1.

[0346] Fifth step: To a solution of 4-((4-((4-((tert-butoxycarbonyl)amino)-2-fluoro-5-(3- hydroxypentan-3-yl)benzamide)methyl)-2,6-dichlorophenoxy)methyl)methyl ester (47-6, 180 mg) in dichloromethane (4 mL) was added trifluoroacetic acid (1 mL) at 0 °C, and the reaction was carried out at 0 °C for 8 hours. The reaction mixture was quenched with saturated sodium bicarbonate (10 mL) and extracted with dichloromethane (15 mL*2). The organic phase was concentrated and purified by reverse phase (acetonitrile / 0.03% formic acid in water) to give compound 47-7 (120 mg). LCMS (ESI) [M-H] - = 454.9.

[0347] Step 6: To a mixture of methyl 4-((4-amino-2-fluoro-5-(3-hydroxypentan-3-yl)benzamide)methyl)- 2,6-dichlorobenzoate (47-7, 120 mg) in methanol (2 mL) and water (1 mL) was added lithium hydroxide (550.51 mg). The reaction mixture was heated to 100 °C and stirred for 1 h. The reaction mixture was cooled to room temperature, concentrated under reduced pressure, and then adjusted to pH 5 with 1 M hydrochloric acid solution. The mixture was extracted with ethyl acetate (15 mL*3), and the combined organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 47-8 (74 mg). LCMS (ESI) [M-H] - = 441.2.

[0348] Step 7: To a solution of benzyl (2S)-2-amino-3-(3-methylsulfonylphenyl)propanoate hydrochloride (62.88 mg) in N,N-dimethylformamide (2 mL) was added 4-((4-amino-2-fluoro-5-(3- hydroxypentan-3-yl)phenylamino)methyl)-2,6-dichlorobenzoic acid (47-8, 75.36 mg), ethyl bis(prop-2-yl)amine (131.82 mg), and HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate) (96.96 mg). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was directly purified by reverse phase separation (C18, acetonitrile / 0.1% formic acid in water system, 70% acetonitrile to elute the product) to give compound 47-10 (74 mg). LCMS (ESI) [M+H-H2O] + = 742.0.

[0349] Step 8: To a solution of benzyl (2S)-2-(4-((4-amino-2-fluoro-5-(3-hydroxypentan-3- yl)benzamide)methyl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propanoate (47-10, 74 mg) in tetrahydrofuran (1 mL) and water (0.2 mL) was added sodium hydroxide (19.51 mg). The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure, and then adjusted to pH 5-6 with 1 M hydrochloric acid. The mixture was purified by preparative separation (acetonitrile / 0.03% formic acid in water) to give compound II-80 (30.77 mg). LCMS (ESI) [M+H-H2O] + = 650.0; 1H NMR (400 MHz, DMSO-d6) δ 12.83 (s, 1H), 9.02 (d, J = 8.0 Hz, 1H), 8.24 (d, J = 8.0 Hz, 1H), 7.86 (s, 1H), 7.76 (d, J = 8.0 Hz, 1H), 7.67 (d, J = 4.0 Hz, 1H), 7.56 (t, J = 8.0 Hz, 1H), 7.35 - 7.33 (m, 3H), 6.37 (d, J = 12.0 Hz, 1H), 6.24 (s, 2H), 4.99 (s, 1H), 4.78 - 4.70 (m, 1H), 4.37 (d, J = 4.0 Hz, 2H), 3.30 - 3.25 (m, 1H), 3.13 (s, 3H), 3.06 - 2.96 (m, 1H), 1.83 - 1.72 (m, 4H), 0.71 (t, J = 8.0 Hz, 6H).

[0350] Example 23

[0351] Preparation of 2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-sulfamoylphenyl)propanoic acid (II-81):

[0352] First Step: To concentrated ammonia (101.63 mL) was added 3-(chlorosulfonyl)benzoic acid (48-1, 10 g) in portions under ice bath. After addition, it was stirred at room temperature for 16 hours. The reaction solution was concentrated, and the pH was adjusted to 1 by dropwise addition of concentrated hydrochloric acid. A large amount of solid precipitated, which was filtered, washed with water, and dried to obtain compound 48-2 (6 g). 1 H NMR (400 MHz, DMSO-d6) 13.43 (s, 3H), 8.40 (s, 1H), 8.14 (d, J = 7.6 Hz, 1H), 8.05 (d, J = 8.0 Hz, 1H), 7.74 - 7.70 (m, 1H), 7.51 (s, 2H).

[0353] Second Step: To 3-sulfamoylbenzoic acid (48-2, 5 g) in THF (50 ml, 100.0%) was added borane tetrahydrofuran (1M in THF, 75 ml) dropwise under ice bath. After dropwise addition, it was stirred at room temperature for 16 hours. The reaction solution was carefully quenched by addition of methanol and hydrochloric acid, stirred at room temperature for 30 minutes, concentrated, and extracted with ethyl acetate. The organic phase was dried and concentrated to obtain compound 48-3 (4 g). LCMS: [M+H] = 188.0. +

[0354] ​Step 3: Dissolve 3-(hydroxymethyl)benzenesulfonamide (48-3, 1 g) in acetone (50 mL, 100.0%) and add manganese dioxide (4.64 g). Stir at 60 °C for 16 h. Cool the reaction to room temperature and filter. Concentrate the filtrate to give compound 48-4 (600 mg). 1 HNMR (400 MHz, DMSO-d6) 10.11 (s, 1H), 8.33 (t, J = 1.6 Hz, 1H), 8.16-8.11 (m, 2H), 7.85-7.81 (m, 1H), 7.55 (s, 2H).

[0355] Step 4: Add benzyl carbamate (48-10, 10 g) and 2,2-dihydroxyacetic acid (6.7 g) to diethyl ether (50 mL, 100.0%). Stir the reaction at room temperature for 16 h. Filter the reaction and wash the filter cake with diethyl ether. Remove the solvent from the solid to give compound 48-12 (4 g). 1 HNMR (400 MHz, DMSO-d6) 12.82 (br, 1H), 8.13 (d, J = 8.8 Hz, 1H), 7.39-7.29 (m, 5H), 6.25 (br, 1H), 5.21 (d, J = 8.8 Hz, 1H), 5.05 (s, 2H).

[0356] Step 5: Dissolve 2-((benzyloxy carbonyl)amino)-2-hydroxyacetic acid (48-12, 4 g) in methanol (50 mL, 100.0%) and add concentrated sulfuric acid (0.44 mL). Stir the reaction at room temperature for 16 h. Wash the reaction with saturated sodium bicarbonate and extract with ethyl acetate. Dry and concentrate the organic phase to give compound 48-13 (4 g). 1 HNMR (400 MHz, CDCl3) 7.37-7.32 (m, 5H), 5.86 (s, 1H), 5.36 (d, J = 9.6 Hz, 1H), 5.16 (s, 2H), 3.81 (s, 3H), 3.47 (s, 3H).

[0357] Step 6: To a solution of methyl 2-(((benzyloxy)carbonyl)amino)-2-methoxyacetate (48-13, 3.1 g) in toluene (30 mL, 100.0%) add phosphorus trichloride (1.85 g) and stir the reaction at 70 °C for 16 h. Cool the reaction to room temperature and add trimethyl phosphite (1.67 g). Stir the reaction at 70 °C for 16 h. Concentrate the reaction and dissolve in ethyl acetate. Wash with saturated aqueous sodium carbonate and dry and concentrate the organic phase to give compound 48-5 (1.5 g). LCMS (ESI) [M+H] = 332.2. + HNMR (400 MHz, CDCl3) 7.37-7.32 (m, 5H), 5.86 (s, 1H), 5.36 (d, J = 9.6 Hz, 1H), 5.16 (s, 2H), 3.81 (s, 3H), 3.47 (s, 3H).

[0358] Step 7: Dimethylguanidine (248.15 mg) was added to a solution of methyl 2-(((benzyloxy)carbonyl)amino)-2-(dimethoxyphosphoryl)acetate (48-5, 747.66 mg) in dichloromethane (10 mL). After the reaction was stirred at room temperature for 30 minutes, it was cooled to -30 °C and 3-formylbenzenesulfonamide (48-4, 380 mg) was added in portions. The reaction was slowly warmed to room temperature and stirred for 16 hours. The reaction was washed with aqueous potassium bisulfate solution, the organic phase was dried and concentrated, slurried in dichloromethane, filtered, and the filter cake was washed with dichloromethane. The solid was dried to remove solvent to give compound 48-6 (350 mg). 1 HNMR (400 MHz, DMSO-d6) 9.33 (br, 1H), 8.10 (s, 1H), 7.85-7.81 (m, 2H), 7.62-7.58 (m, 1H), 7.40-7.28 (m, 8H), 5.10 (s, 2H), 3.72 (s, 3H).

[0359] Step 8: To a solution of (E)-methyl 2-((benzyloxy)carbonyl)amino)-3-(3- sulfamoylphenyl)acrylate (48-6, 350 mg) in methanol (5 ml) was added palladium on carbon (35 mg) and the reaction was stirred under a hydrogen balloon at room temperature for 16 hours. The reaction was filtered and concentrated to give compound 48-7 (150 mg). LCMS (ESI) [M+H] + = 259.0.

[0360] Step 9: 4-({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]formamido}methyl)- 2,6-dichlorobenzoic acid (48-8, 80.38 mg,) and HATU (2-(7-azabenzotriazol)-N,N,N',N'- tetramethyluronium hexafluorophosphate) (108.79 mg) were dissolved in DMSO (2 mL, 100.0%). Triethylamine (57.68 mg) and methyl 2-amino-3-(3-sulfamoylphenyl)propanoate (48-7, 49.08 mg) were added sequentially and the reaction was stirred at room temperature for 16 hours. The reaction was purified by reverse phase (FA in H2O / MeCN, 43%) to give compound 48-9 (30 mg). LCMS (ESI) [M-17] + = 637.0.

[0361] Tenth step: Methyl 2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamide)methyl)- 2,6-dichlorobenzamide)-3-(3-sulfamoylphenyl)propanoate (48-9, 30 mg) was dissolved in methanol (0.5 mL), and a solution of lithium hydroxide monohydrate (19.2 mg) in water (0.5 mL) was added. The reaction was stirred at room temperature for 4 hours. The reaction was adjusted to pH = 4 with 1 M aqueous hydrochloric acid solution, and purified by preparative liquid phase (H2O / MeCN formic acid system) to give compound II-81 (7 mg). LCMS (ESI) [M-H] - = 638.9; 1 H NMR (400 MHz, DMSO-d6) δ 12.83 (br, 1H), 8.98 (s, 1H), 8.29 (d, J = 6.0 Hz, 1H), 7.75 (s, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.52 (d, J = 8.0 Hz, 1H), 7.49 - 7.40 (m, 2H), 7.34 (s, 2H), 7.29 (s, 2H), 6.40 (d, J = 14.0 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.64 (s, 1H), 4.38 (d, J = 6.4 Hz, 2H), 3.21 (dd, J = 14.4, 4.8 Hz, 1H), 3.00 (dd, J = 13.6, 9.2 Hz, 1H), 1.48 (s, 6H).

[0362] Example 24

[0363] Preparation of (S)-2-(4-((3-amino-2-(2-hydroxypropan-2-yl)quinolin-7-yl)methyl)-2,6- dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-82):

[0364] First step: Ethyl 3-bromo-2-oxopropanoate (49-2, 10.24 g) was dissolved in ethanol (50 mL), and a solution of pyridine (4 mL) in ethanol (50 mL) was added dropwise. The reaction was stirred at room temperature for 1 hour, and then heated to 60 °C for 2 hours. After cooling to room temperature, 2-amino-4-bromobenzaldehyde (49-1, 10.0 g) and pyridine (12 mL) were added, and the reaction was stirred at 80 °C overnight. Tetrahydropyrrole (10.46 mL) was added, and the reaction was stirred at 85 °C for 4 hours. The reaction was concentrated under reduced pressure, and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to give compound 49-3 (6.1 g). LCMS (ESI) [M+H] + = 297.0.

[0365] Second step: 3-amino-7-bromoquinoline-2-carboxylic acid ethyl ester (49-3, 6 g) was dissolved in super dry THF (120 mL), the reaction solution was purged with nitrogen and then methyl magnesium bromide (3.0 M in THF, 40.4 mL) was added dropwise. After the addition was completed, the reaction solution was allowed to warm to room temperature and stirring was continued for 1 hour. The reaction solution was quenched with saturated ammonium chloride solution (50 mL), the resulting white suspension was filtered through celite, the filtrate was diluted with water (100 mL) and then extracted with ethyl acetate (100 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. Purification was performed by column chromatography (petroleum ether: ethyl acetate = 5: 1) to give compound 49-4 (640 mg). LCMS (ESI) [M+H] + = 281.0.

[0366] Third step: 2-(3-amino-7-bromoquinolin-2-yl)propan-2-ol (49-4, 640 mg) was dissolved in 1,4-dioxane (10 mL), 4,4,5,5-tetramethyl-2-(4,4,5,5-4-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (693.67 mg), potassium carbonate (943.85 mg) and Pd(dppf)Cl2(83.28 mg) were added, the reaction solution was purged with nitrogen three times and stirred at 90 °C for 16 hours. Celite was added to the reaction solution and filtered, the filtrate was concentrated to give crude compound 49-6 (640 mg). LCMS (ESI) [M+H] + = 329.2.

[0367] Fourth step: 2-(3-amino-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinolin-2-yl)propan-2-ol (49-6, 640 mg) was dissolved in 1,4-dioxane (5 mL) and water (1 mL), methyl 2,6-dichloro-4-(chloromethyl)benzoate (494.34 mg), potassium carbonate (808.52 mg) and Pd(dppf)Cl2(114.14 mg) were added, the reaction solution was purged with nitrogen three times and stirred at 90 °C for 16 hours. The reaction solution was filtered through celite, the filtrate was concentrated under reduced pressure and purified by column chromatography (petroleum ether: ethyl acetate = 2: 1) to give compound 49-8 (630 mg). LCMS (ESI) [M+H] + = 419.0.

[0368] Step 5: Methyl 4-((3-amino-2-(2-hydroxypropyl-2-yl)quinoline-7-yl)methyl)-2,6-dichlorobenzoate (49-8, 760 mg) was dissolved in methanol (6 mL) and tetrahydrofuran (3 mL), and lithium hydroxide (3.8 g) was dissolved in water (3 mL). The mixture was added to the reaction solution and reacted at 100 °C for 16 hours. A large amount of lithium hydroxide was removed by filtration, and the solvent was removed by concentration under reduced pressure. The pH was adjusted to approximately 5 with 2M hydrochloric acid aqueous solution (5 mL). The product was purified by reverse-phase separation (C18, acetonitrile / 0.1% formic acid aqueous solution system, 40% acetonitrile content yielded the product) to obtain compound 49-9 (70 mg). LCMS (ESI) [M+H] + =404.6.

[0369] Step 6: Dissolve 4-((3-amino-2-(2-hydroxypropyl-2-yl)quinoline-7-yl)methyl)-2,6-dichlorobenzoic acid (49-9, 50 mg) in N,N-dimethylformamide (2 mL), add HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (70.37 mg) and triethylamine (74.91 mg), and stir at 45 °C for 10 minutes. Add (2S)-2-amino-3-(3-methanesulfonylphenyl)propionate benzyl hydrochloride (50.19 mg), and stir at 45 °C for 2 hours. The reaction solution is directly purified by reverse-phase separation (C18, acetonitrile / 0.1% formic acid aqueous solution system, 40% acetonitrile content yields the product) to give compound 49-11 (70 mg). LCMS (ESI) [M+H] + =720.1.

[0370] Step 7: Benzyl (S)-2-(4-(3-amino-2-(2-hydroxypropyl-2-yl)quinoline-7-yl)methyl)-2,6-dichlorobenzamido)-3-(3-(methanesulfonyl)phenyl)propionate (49-11, 70 mg) was dissolved in tetrahydrofuran (2 mL), and sodium hydroxide (19.43 mg) was dissolved in water (0.4 mL). The mixture was added to the reaction solution and stirred at room temperature for 3 hours. The reaction solution was purified by prep-HPLC (C18, 10 mmol / L FA in water, MeCN) to obtain compound II-82 (30 mg). LCMS (ESI) [M+H] + =630.0; 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.80 (s, 1H), 7.71 (d, J = 7.6 Hz, 1H), 7.65 - 7.58 (m, 2H), 7.53 - 7.49 (m, 2H), 7.37 (s, 2H), 7.23 (d, J = 8.4 Hz, 1H), 7.17 (s, 1H), 5.84 (s, 2H), 5.71 (s, 1H), 4.48 (s, 1H), 4.02 (s, 2H), 3.27 - 3.23 (m, 1H), 3.07 - 3.02 (m, 1H), 1.59 (s, 6H).

[0371] Example 25

[0372] Preparation of (S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(dimethylphosphoryl)phenyl)propanoic acid (II-83):

[0373] First Step: (2S)-3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid (50-1, 10 g) was dissolved in N,N-dimethylformamide (100 mL), and iodomethane (6.19 g) and sodium bicarbonate (4.88 g) were added. The reaction solution was stirred at room temperature for 16 hours. The reaction solution was concentrated, and compound 50-2 (9.6 g) was isolated by column chromatography (petroleum ether: ethyl acetate = 5:1). LCMS (ESI) [M+H-t-Bu] = 301.7. +

[0374] Second Step: In a microwave tube, (S)-methyl 3-(3-bromophenyl)-2-((tert- butyloxycarbonyl)amino)propanoate (50-2, 2 g) was dissolved in N,N-dimethylformamide (20 mL), and methyl phosphonous acid methane (479.33 mg), palladium acetate (62.67 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (193.83 mg), and potassium phosphate (1.3 g) were added. The reaction solution was stirred at 150 °C in a microwave reactor for 30 minutes. The reaction solution was separated by reverse phase (C18, acetonitrile / 0.1% trifluoroacetic acid aqueous system, 35% acetonitrile content to produce the product) to obtain compound 50-3 (570 mg). LCMS (ESI) [M+H] = 356.1. +

[0375] ​​Step 3: (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(3- (dimethylphosphoryl)phenyl)propanoate (50-3, 570 mg) was dissolved in ethyl acetate solution of hydrochloric acid (4.0 M, 8 mL) and stirred at room temperature for 2 hours. The solvent was removed by concentration under reduced pressure, and the residue was slurried with ethyl ether (10 mL) and filtered to give compound 50-4 (400 mg). LCMS (ESI) [M+H] + = 255.9.

[0376] Step 4: 4-({[4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl]formamido}methyl)- 2,6-dichlorobenzoic acid (50-5, 357.9 mg) was dissolved in N,N- dimethylformamide (3 mL), and HATU (2-(7-azabenzotriazol-1- yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (446.89 mg) and triethylamine (396.43 mg) were added, and stirred at room temperature for 20 minutes. (S)-methyl 2-amino-3-(3-(dimethylphosphoryl)phenyl)propanoate (50-4, 200 mg) was added, and stirred at room temperature for 1 hour. The reaction mixture was directly purified by reverse phase separation (C18, acetonitrile / 0.1% formic acid aqueous system, 40% acetonitrile content to give the product) to give compound 50-6 (40 mg). LCMS (ESI) [M+H] + = 652.3.

[0377] Step 5: (S)-methyl 2-(4-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido-3-(3-(dimethylphosphoryl)phenyl)propanoate (50-6, 40 mg) was dissolved in tetrahydrofuran (1 mL). Sodium hydroxide (12.26 mg) was dissolved in water (0.2 mL), and added to the reaction mixture. The mixture was stirred at room temperature for 2 hours. The reaction mixture was purified by prep-HPLC separation (C18, acetonitrile / 0.1% formic acid aqueous system) to give compound II-83 (24 mg). LCMS (ESI) [M+H-H2O] + = 620.3; 1H NMR (400 MHz, DMSO-d6) δ 12.97 (s, 1H), 8.77 (s, 1H), 7.29 - 7.23 (m, 1H), 7.68 (d, J = 12.0 Hz, 1H), 7.59 - 7.54 (m, 1H), 7.48 - 7.37 (m, 3H), 7.32 (s, 2H), 6.40 (d, J = 14.0 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.64 (s, 1H), 4.37 (d, J = 5.6 Hz, 2H), 3.22 (dd, J = 14.0, 4.4 Hz, 1H), 3.00 - 2.95 (m, 1H), 1.61 (s, 3H), 1.58 (s, 3H), 1.48 (s, 6H).

[0378] Example 26

[0379] Preparation of (S)-2-(4-(2-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl)pyrimidin-4-yl)- 2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-84):

[0380] First Step: 4-bromo-2,6-dichlorobenzoic acid (51-1, 20 g) was dissolved in N,N- dimethylformamide (200 mL) at room temperature, potassium carbonate (30.72 g) was added at room temperature, and iodomethane (21.04 g) was added. The reaction was stirred at 25 °C for 4 hours, and the reaction system was quenched with an aqueous solution (200 mL), extracted with ethyl acetate (200 mL) three times. The organic phase was combined and dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by flash column chromatography (Silica, petroleum ether / tetrahydrofuran = 10 / 1) to obtain compound 51-2 (20 g). 1 H NMR (400 MHz, CDCl3) δ 7.50 (s, 2H), 3.97 (s, 3H).

[0381] Second step: 4-bromo-2,6-dichlorobenzoic acid methyl ester (51-2, 19.5 g) was dissolved in 1,4-dioxane (260 mL) at room temperature, 4,4,5,5-tetramethyl-2-(4,4,5,5-4-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane (20.93 g), potassium acetate (20.22 g), 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (5.03 g) were added at room temperature, and the mixture was stirred at 80 °C for 16 hours under nitrogen protection. Water (200 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (200 mL) three times. The organic phase was combined and washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by flash chromatography (Silica, petroleum ether / tetrahydrofuran = 8 / 1) to give compound 51-3 (22 g). LCMS (ESI) [M+H] + = 331.0; 1 H NMR (400 MHz, CDCl3) δ 7.72 (s, 2H), 3.97 (s, 3H), 1.34 (s, 12H).

[0382] Third step: 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (2.21 g) was added to a mixture of 2,6-dichloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoic acid methyl ester (51-3, 10 g), 2,4-dichloropyrimidine (9 g), potassium carbonate (8.35 g) in 1,4-dioxane (100 mL) and water (20 mL) under nitrogen atmosphere, and the mixture was stirred at 45 °C for 6 hours. The mixture was cooled to room temperature, filtered, and the filtrate was concentrated. The residue was purified by flash chromatography (Silica gel, petroleum ether / ethyl acetate = 10 / 1) to give compound 51-5 (6.82 g). LCMS (ESI) [M+H+ACN] + = 358.0; 1 H NMR (400 MHz, CDCl3) δ 8.74 (d, J = 5.2 Hz, 1H), 8.06 (s, 2H), 7.64 (d, J = 5.2 Hz, 1H), 4.02 (s, 3H).

[0383] Fourth step: under nitrogen atmosphere, 4,4,5,5-tetramethyl-2-(4,4,5,5-4-tetramethyl-1,3,2- dioxaborolan-2-yl)-1,3,2-dioxaborolane (10.94 g) was added to tert-butyl N-[4-bromo-5-fluoro- 2-(2-hydroxypropan-2-yl)phenyl]carbamate (51-6, 6 g), potassium acetate (5.07 g), 1,1- bis(diphenylphosphino)ferrocene palladium dichloride (1.26 g) in 1,4-dioxane (100 mL), 90 °C for 16 hours, cooled to room temperature, the reaction mixture was filtered, the filtrate was concentrated, the residue was separated and purified by flash chromatography (Silica gel, petroleum ether / ethyl acetate = 85% / 15%) to give compound 51-7 (2.8 g). LCMS (ESI) [M+H-H2O-t-Bu+ACN] = 363.1. + 1 H NMR (400 MHz, DMSO-d6) δ 10.07 (s, 1H), 7.82 (d, J = 12.3 Hz, 1H), 7.43 (d, J = 6.8 Hz, 1H), 6.16 (s, 1H), 1.50 (s, 6H), 1.47 (s, 9H), 1.28 (s, 12H).

[0384] Fifth step: under nitrogen atmosphere, 1,1-bis(diphenylphosphino)ferrocene palladium dichloride (0.35 g) was added to methyl 2,6-dichloro-4-(2-chloropyrimidin-4-yl)benzoate (1.5 g), tert-butyl (5-fluoro-2-(2-hydroxypropan-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)carbamate (51-7, 2.8 g), potassium carbonate (1957.05 mg) in 1,4-dioxane (32 mL) and water (8 mL), 90 °C for 16 hours. Cooled to room temperature, the reaction mixture was filtered, the filtrate was concentrated, the residue was separated and purified by flash chromatography (Silica gel, petroleum ether / ethyl acetate = 10 / 1) to give compound 51-8 (2.2 g). LCMS (ESI) [M+H] + = 550.1.

[0385] ​Step 6: Lithium hydroxide (4.21 g) was added to methyl 4-(2-(4-((tert-butoxycarbonyl)amino)-2-fluoro-5-(2-hydroxypropyl-2-yl)phenyl)pyrimidin-4-yl)-2,6-dichlorobenzoate (51-8, 1.94 g) in methanol (20 mL) and water (5 mL). The reaction was carried out at 100 °C for 2 hours. The reaction solution was concentrated, and the residue was added to water (10 mL). The pH was adjusted to 7 with 1 M hydrochloric acid, and the residue was extracted with ethyl acetate (10 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by reverse-phase chromatography (C18, 0.1% FA in water / MeCN = 50% / 50%) to obtain compound 51-9 (600 mg). LCMS (ESI) [M+H] + =462.0.

[0386] Step 7: Lithium hydroxide (1891.09 mg) was added to 2,6-dichloro-4-(2-(7-fluoro-4,4-dimethyl-2-oxo-1,4-dihydro-2H-benzo[d][1,3]oxazin-6-yl)pyrimidin-4-yl)benzoic acid (51-9, 730 mg) in methanol (32 mL) and water (16 mL). The reaction was carried out at 100 °C for 16 hours. The reaction solution was concentrated to remove methanol, and the pH was adjusted to 7 with 1 M hydrochloric acid. The solution was concentrated again, and the residue was analyzed by prep-HPLC (C18, 0.1% NH4HCO3 in water / MeCN) to give compound 51-10 (80 mg). LCMS (ESI) [M+H] + =436.0.

[0387] Step 8: Add HATU (2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate) (91.51 mg) to 4-(2-(4-amino-2-fluoro-5-(2-hydroxypropyl-2-yl)phenyl)pyrimidin-4-yl)-2,6-dichlorobenzoic acid (51-10, 70 mg, 0.16 mmol, 1 eq), 2-amino-3-(3-methanesulfonylphenyl)propionate benzyl ester hydrochloride (89.02 mg), N,N-diisopropylethylamine (62.21 mg, 0.48 mmol, 3 eq) in N,N-dimethylformamide (10 mL), and react at 25°C for 1 hour. The reaction mixture was added to water (20 mL), extracted with ethyl acetate (15 mL x 3), the organic phases were combined, washed with saturated brine (15 mL x 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 compound 51-12 (100 mg). LCMS (ESI) [M+H] + =751.1.

[0388] Ninth step: 1M aqueous sodium hydroxide solution (5 mL) was added to (S)-2-(4-(2-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl)pyrimidin-4-yl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (51-12, 90 mg) in tetrahydrofuran (5 mL) at 25 °C for 1 h. The pH was adjusted to 7 with saturated aqueous citric acid solution, the reaction was concentrated, and the residue was purified by prep-HPLC (C18, 10 mmol NH4HCO3 in water / MeCN) to give compound II-84 (29.12 mg). LCMS (ESI) [M+H] + = 661.0; 1 H NMR (400 MHz, DMSO-d6) δ 8.91 (d, J = 5.3 Hz, 1H), 8.58 (s, 1H), 8.28 (s, 2H), 7.99 (dd, J = 13.3, 7.1 Hz, 2H), 7.85 (s, 1H), 7.75 (d, J = 7.8 Hz, 1H), 7.67 (d, J = 7.7 Hz, 1H), 7.54 (t, J = 7.7 Hz, 1H), 6.51 (d, J = 14.0 Hz, 1H), 6.23 (s, 2H), 5.41 (s, 1H), 4.56 (d, J = 4.8 Hz, 1H), 3.33 (s, 1H), 3.16 - 3.10 (m, 4H), 1.55 (s, 6H).

[0389] Example 27

[0390] Preparation of (S)-2-(4-(6-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl)pyridin-2-yl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-85):

[0391] First step: iodomethane (16679.8 mg) was slowly added to a solution of 2-amino-5-bromo-4-fluorobenzoic acid (52-1, 25 g) and sodium bicarbonate (13462.18 mg) in N,N-dimethylformamide (250 mL) at 0 °C, after the addition was completed, the temperature was raised to 30 °C and the reaction was allowed to proceed for 16 h. Water (250 mL) was added to the reaction, and the mixture was extracted with ethyl acetate (200 mL), the organic phases were combined, washed with saturated brine (300 mL*3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by normal phase silica gel (petroleum ether: ethyl acetate = 10:1) to give compound 52-2 (17.4 g). LCMS (ESI) [M+H] + = 248.0.

[0392] Second step: To a solution of methyl 2-amino-5-bromo-4-fluorobenzoate (52-2, 8 g) in tetrahydrofuran (100 mL) was added 3M methyl magnesium bromide solution (64.5 mL) slowly at 0 °C, after the addition was completed, the reaction was allowed to warm to 25 °C for 1 hour. The reaction was quenched by the addition of saturated aqueous ammonium chloride solution (100 mL) under ice-bath, extracted with ethyl acetate (50 mL*3), the organic phase was combined and washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried and concentrated to give the crude compound 52-3 (8001.29 mg). LCMS (ESI) [M+H-H2O] = 230.2. +

[0393] Third step: To a solution of 2-(2-amino-5-bromo-4-fluorophenyl)propan-2-ol (52-3, 350 mg) and 2-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (405.47 mg) in tetrahydrofuran (1 mL) and water (1 mL) was added triethylamine (428.27 mg) and [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium (91.95 mg) under nitrogen atmosphere, the reaction was carried out at 50 °C for 2 hours. The reaction was filtered, then diluted with water (10 mL), extracted with ethyl acetate (10 mL*3), the organic phase was combined and washed with saturated sodium chloride solution (10 mL*3), the organic phase was dried and concentrated to give the crude product, which was purified by reverse phase separation (C18, 10 mmol / L FA in water, MeCN) to give compound 52-5 (300 mg). LCMS (ESI) [M+H-H2O] = 263.0. +

[0394] Fourth step: To a solution of 2-[2-amino-5-(6-chloropyridin-2-yl)-4-fluorophenyl]propan-2-ol (52-5, 300.38 mg) and methyl 2,6-dichloro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (425 mg) in 1,4-dioxane (5 mL) and water (1 mL) was added potassium carbonate (443.65 mg) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (81.62 mg) under nitrogen atmosphere, the reaction was carried out at 110 °C for 3 hours. The reaction was filtered, then diluted with water (10 mL), extracted with ethyl acetate (5 mL*3), the organic phase was combined and washed with saturated sodium chloride solution (20 mL*3), the organic phase was dried and concentrated to give the crude product, which was purified by reverse phase separation (C18, 10 mmol / L FA in water, MeCN) to give compound 52-7 (300 mg). LCMS (ESI) [M+H] + ​​= 449.1.

[0395] Step 5: Lithium iodide (714.98 mg) was added to methyl 4-(6-(4-amino-2-fluoro-5-(2- hydroxypropan-2-yl)phenyl)pyridin-2-yl)-2,6-dichlorobenzoate (52-7, 400 mg) in pyridine (10 mL) at 125 °C for 3 hours. Cool to room temperature, compound 52-8 (270 mg) was obtained by reverse phase system (C18, 0.1% FA in water / ACN = 30% / 70%). LCMS (ESI) [M+H] + = 435.0; 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 2H), 8.02 - 7.89 (m, 3H), 7.71 (d, J = 7.3 Hz, 1H), 6.51 (d, J = 14.3 Hz, 1H), 6.06 (s, 1H), 5.39 (s, 1H), 1.55 (s, 6H).

[0396] Step 6: HATU (2-(7-Azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (326.84 mg) was added to 4-(6-(4-amino-2-fluoro-5-(2-hydroxypropan-2- yl)phenyl)pyridin-2-yl)-2,6-dichlorobenzoic acid (52-8, 250 mg), benzyl 2-amino-3-(3- methanesulfonylphenyl)propanoate hydrochloride (317.92 mg), ethyl bis(2-propyl)amine (222.18 mg) in dimethylformamide (10 mL) at 25 °C for 1 hour. The reaction was added water (20 mL), extracted with ethyl acetate (15 mL*3), the combined organic phase was washed with saturated brine (20 mL*3), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, the residue was separated and purified by flash chromatography (Silica gel, dichloromethane / methanol = 10 / 1) to give compound 52-10 (420 mg). LCMS (ESI) [M+H] + = 750.1.

[0397] Seventh Step: 1M aqueous sodium hydroxide solution (6 mL) was added to (S)-2-(4-(6-(4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)phenyl)pyridin-2-yl)-2,6-dichlorobenzamide)-3-(3-(methylsulfonyl)phenyl)propionic acid benzyl ester (52-10, 420 mg) in tetrahydrofuran (6 mL) at 25 °C for 1 h. The pH was adjusted to 7 with 1M hydrochloric acid, the reaction was concentrated, and the residue was purified by prep-HPLC (C18, 10 mmol NH4HCO3 in water / MeCN) to give compound II-85 (65.24 mg). LCMS (ESI) [M+H] + = 660.1; 1 H NMR (400 MHz, DMSO-d6) δ 8.63 (s, 1H), 8.18 (s, 2H), 7.96 - 7.86 (m, 4H), 7.76 (d, J = 7.8 Hz, 1H), 7.68 (d, J = 7.8 Hz, 2H), 7.55 (t, J = 7.7 Hz, 1H), 6.51 (d, J = 14.2 Hz, 1H), 6.05 (s, 2H), 5.38 (s, 1H), 4.59 (d, J = 5.4 Hz, 1H), 3.33 - 3.30 (m, 1H), 3.16 - 3.08 (m, 4H), 1.57 (s, 6H).

[0398] Example 28

[0399] Preparation of (S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamide)methyl)-2,6-dichlorobenzamide)-3-(3-aminosulfonylphenyl)propanoic acid (II-86):

[0400] First Step: HATU (2-(7-Azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (883.26 mg) was added to (S)-2-amino-3-(3-sulfonamidophenyl)propanoic acid methyl ester (53-1, 400 mg), 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzoic acid (53-2, 771.67 mg), ethyl bis(2-propyl)amine (600.44 mg) in N,N-dimethylformamide (10 mL) and reacted at 25 °C for 1 h. The reaction solution was added to water (20 mL), extracted with ethyl acetate (15 mL*3), the combined organic phase was washed with saturated brine (20 mL*3), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and the residue was separated and purified by flash chromatography (Silica gel, ethyl acetate / methanol = 10 / 1) to obtain compound 53-3 (230 mg). LCMS (ESI) [M+H-H2O]=637.1. +

[0401] Second Step: Lithium hydroxide (84.03 mg) was added to (S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzamido)-3-(3- aminosulfonylphenyl)propanoic acid methyl ester (53-3, 230 mg) in methanol (5 mL) and water (5 mL) and reacted at 25 °C for 1 h. The pH was adjusted to 7 with 1M hydrochloric acid, the reaction solution was concentrated, and the residue was obtained by prep-HPLC (C18, 0.1% NH4HCO3 in water / MeCN) to obtain compound II-86 (38.95 mg). LCMS (ESI) [M+H-H2O]=642.1; + 1 H NMR (400 MHz, DMSO-d6) δ 8.39-8.20 (m, 2H), 7.72 (s, 1H), 7.63 (d, J=7.9 Hz, 1H), 7.51 (d, J=7.7 Hz, 1H), 7.45-7.39 (m, 2H), 7.33 (s, 2H), 6.40 (d, J=13.9 Hz, 1H), 6.20 (s, 2H), 5.36 (s, 1H), 4.38 (d, J=5.8 Hz, 3H), 3.23-3.16 (m, 1H), 3.11-3.03 (m, 1H), 1.48 (s, 6H).

[0402] Example 29

[0403] ​​Preparation of (R)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamide)methyl)- 2,6-dichlorobenzamide)-3-(3-aminosulfonylphenyl)propanoic acid (II-87):

[0404] First step: (R)-methyl 2-amino-3-(3-aminosulfonylphenyl)propanoate (54-1, 400 mg) was dissolved in N,N-dimethylformamide (8 mL), 4-((4-amino-2-fluoro-5-(2- hydroxypropan-2-yl)benzamide)methyl)-2,6-dichlorobenzoic acid (54-2, 771.67 mg), HATU (2-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate) (883.26 mg), ethyl bis(2-propyl)amine (600.44 mg) were added successively, the reaction system was stirred at room temperature for 1 hour, water (25 mL) was added, extraction was performed with ethyl acetate (25 mL*2), the combined organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was separated by silica gel column (dichloromethane:methanol = 10:1), and then purified by silica gel column (ethyl acetate:methanol = 10:1) to give compound 54-3 (390 mg). LCMS (ESI) [M+H-H20] + = 637.1.

[0405] Second step: (R)-methyl 2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamide)methyl)- 2,6-dichlorobenzamido)-3-(3-aminosulfonylphenyl)propanoate (54-3, 400 mg) was dissolved in methanol (4 mL) and water (4 mL), lithium hydroxide (146.14 mg) was added, the reaction system was stirred at room temperature for 1 hour, the reaction solution was adjusted to neutral pH with 1M hydrochloric acid, concentrated to give a crude product, which was separated and purified by prep-HPLC (0.1% FA in water / ACN), and then separated and purified by prep-HPLC (10 mmol NH4HCO3 in water / ACN) to give compound II-87 (55.63 mg). LCMS (ESI) [M+H-H20] + = 623.1; 1H NMR (400 MHz, DMSO-d6) δ 8.66 (s, 1H), 8.33 - 8.25 (m, 1H), 7.74 (s, 1H), 7.65 (d, J = 7.8 Hz, 1H), 7.52 (d, J = 7.8 Hz, 1H), 7.47 - 7.42 (m, 2H), 7.36 - 7.23 (m, 4H), 6.40 (d, J = 13.9 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.51 (s, 1H), 4.38 (d, J = 5.7 Hz, 2H), 3.23 - 3.17 (m, 1H), 3.07 - 3.01 (m, 1H), 1.48 (s, 6H).

[0406] Example 30

[0407] Preparation of (2S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)- 2,6-dichlorobenzamido)-3-(3-(S-methanesulfonylimino)phenyl)propanoic acid (II-88):

[0408] First step: (S)-3-(3-bromophenyl)-2-((tert-butoxycarbonyl)amino)propanoic acid (55-1, 5 g) was dissolved in tetrahydrofuran (100 mL), 2.5M n-butyllithium (17.45 mL) hexane solution was added dropwise at -78°C, and the mixture was stirred for 30 minutes. Dimethyl disulfide (2736.78 mg) tetrahydrofuran solution (10 mL) was added dropwise, and the mixture was stirred for 2 hours. Saturated ammonium chloride (100 mL) was added, and the mixture was extracted with ethyl acetate (80 mL*3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified by silica gel column (petroleum ether: ethyl acetate = 1:1) to give compound 55-2 (2.8 g). LCMS (ESI) [M+H-Boc] = 212.1. +

[0409] Second step: (S)-2-((tert-butoxycarbonyl)amino)-3-(3-(methylthio)phenyl)propanoic acid (55-2, 2.8 g) was dissolved in acetone (28 mL), and iodomethane (1.66 g) and potassium carbonate (1.86 g) were added successively. The mixture was stirred at 60°C for 12 hours. The reaction solution was filtered, and the filtrate was concentrated to give a crude product. The crude product was purified by silica gel column (petroleum ether: ethyl acetate = 2:1) to give compound 55-3 (2.7 g). LCMS (ESI) [M+H-Boc] = 226.2; + 1 ​​H NMR (400 MHz, CDC13) δ 7.21 (t, J = 7.7 Hz, 1H), 7.15 - 7.12 (m, 1H), 7.00 (s, 1H), 6.89 (d, J = 7.4 Hz, 1H), 4.66 - 4.47 (m, 1H), 3.72 (s, 3H), 3.14 - 2.97 (m, 2H), 2.47 (s, 3H), 1.42 (s, 9H).

[0410] Step 3: (S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(3-(methylthio)phenyl)propanoate (55-3, 2.7 g) was dissolved in methanol (30 mL), and ammonium carbonate (1.59 g) and (diacetoxyiodo)benzene (6.68 g) were added successively. The reaction system was stirred at room temperature for 2 hours. The reaction solution was concentrated, dichloromethane (50 mL) was added, and the filtrate was concentrated to obtain a crude product. The crude product was separated and purified by silica gel column (dichloromethane:methanol = 10:1) to obtain compound 55-4 (2.2 g). LCMS (ESI) [M+H] + = 357.1; 1 H NMR (400 MHz, CDC13) δ 7.21 (t, J = 7.7 Hz, 1H), 7.15 - 7.12 (m, 1H), 7.00 (s, 1H), 6.89 (d, J = 7.4 Hz, 1H), 4.66 - 4.47 (m, 1H), 3.72 (s, 3H), 3.14 - 2.97 (m, 2H), 2.47 (s, 3H), 1.42 (s, 9H).

[0411] Step 4: (2S)-methyl 2-((tert-butoxycarbonyl)amino)-3-(3-(S-methylsulfonimidoyl)phenyl)propanoate (55-4, 800 mg) was dissolved in ethyl acetate (8 mL), and 4M hydrochloric acid ethyl acetate solution (8 mL) was added. The reaction system was stirred at room temperature for 2 hours. The reaction solution was filtered and dried to obtain compound 55-5 (550 mg). LCMS (ESI) [M+H] + = 257.2; 1 H NMR (400 MHz, DMSO-d6) δ 8.85 (s, 3H), 8.16 (d, J = 4.5 Hz, 1H), 8.11 - 8.06 (m, 1H), 7.86 - 7.79 (m, 2H), 4.49 (s, 1H), 3.85 (d, J = 3.1 Hz, 3H), 3.78 (d, J = 3.2 Hz, 3H), 3.43 - 3.36 (m, 2H).

[0412] Step 5: (2S)-2-Amino-3-(3-(S-methanesulfonylimino)phenyl)propanoic acid methyl ester hydrochloride (55-5, 550 mg) was dissolved in N,N-dimethylformamide (10 mL), 4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6-dichlorobenzoic acid (936.06 mg), HATU (2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate) (1071.42 mg), ethyl bis(2-propyl)amine (728.35 mg) were added successively, the reaction system was stirred at room temperature for 1 hour, water (30 mL) was added, extracted with ethyl acetate (35 mL*2), the combined organic phase was washed with saturated brine (20 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was separated by silica gel column (dichloromethane:methanol = 10:1), and then purified by silica gel column (ethyl acetate:methanol = 10:1) to give compound 55-7 (470 mg). LCMS (ESI) [M+H-H20] = 653.2. +

[0413] Step 6: (2S)-2-(4-((4-amino-2-fluoro-5-(2-hydroxypropan-2-yl)benzamido)methyl)-2,6- dichlorobenzamido)-3-(3-(S-methanesulfonylimino)phenyl)propanoic acid methyl ester (55-7, 400 mg) was dissolved in methanol (4 mL) and water (4 mL), lithium hydroxide (146.58 mg) was added, the reaction system was stirred at room temperature for 1 hour, the reaction solution was adjusted to neutral pH with 1M hydrochloric acid, and concentrated to give a crude product, which was separated and purified by prep-HPLC (0.1% FA in water / ACN) to give compound II-88 (297.54 mg). LCMS (ESI) [M+H] + = 639.2; 1 H NMR (400 MHz, DMSO-d6) δ 12.88 (s, 1H), 9.02 (d, J = 7.9 Hz, 1H), 8.33 - 8.24 (m, 1H), 7.87 (s, 1H), 7.80 - 7.74 (m, 1H), 7.62 - 7.55 (m, 1H), 7.50 (t, J = 7.7 Hz, 1H), 7.43 (d, J = 8.7 Hz, 1H), 7.32 (s, 2H), 6.40 (d, J = 14.0 Hz, 1H), 6.20 (s, 2H), 5.35 (s, 1H), 4.79 - 4.66 (m, 1H), 4.37 (d, J = 5.9 Hz, 2H), 4.10 (s, 1H), 4.10 (s, 1H), 3.28 - 3.23 (m, 1H), 3.03 - 2.97 (m, 4H), 1.48 (s, 6H).

[0414] ​Example 31

[0415] Preparation of (S)-2-(4-((4-amino-5-(2-hydroxypropan-2-yl)thiophene-3- carboxamido)methyl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid (II-89):

[0416] First step: 4-(((tert-butoxycarbonyl)amino)methyl)-2,6-dichlorobenzoic acid (56-1, 1.0 g), (S)-2-amino-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (1.39 g), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.43 g) were dissolved in N,N-dimethylformamide (10 mL), ethyl bis(2-propyl)amine (1.41 g) was added, the reaction system was stirred at room temperature for 4 hours, water (30 mL) was added, extracted with ethyl acetate (30 mL*3), the combined organic phase was washed with saturated brine (30 mL*2), dried over anhydrous sodium sulfate, filtered and concentrated to give compound 56-2 (1.9 g). LCMS (ESI) [M+H-t-Bu] + = 579.0.

[0417] Second step: (S)-2-(4-((((tert-butoxycarbonyl)amino)methyl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (56-2, 1.9 g) was dissolved in ethyl acetate (40 mL), 4M hydrochloric acid ethyl acetate (10 mL) was added, the reaction system was stirred at room temperature for 2 hours, the reaction liquid was filtered, and the filter cake was dried to give compound 56-3 (1.5 g). LCMS (ESI) [M+H] + = 535.2.

[0418] Third step: 3-amino-4-bromothiophene-2-carboxylic acid methyl ester (56-4, 10 g) was dissolved in tetrahydrofuran (100 mL), 3M methyl magnesium bromide (141.19 mL) in 2-methyltetrahydrofuran was added dropwise in an ice bath, after the addition was completed, the stirring was continued at 45°C for 16 hours, the reaction liquid was poured into a mixture of ice and saturated ammonium chloride until it was completely quenched, extracted with ethyl acetate (100 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give a crude product, which was purified by silica gel column (petroleum ether: ethyl acetate = 2:1) to give compound 56-5 (3.6 g). LCMS (ESI) [M+H-H2O] + = 219.9; 1 H NMR (400 MHz, DMSO-d6) δ 7.29 (s, 1H), 5.45 (s, 1H), 4.59 (s, 2H), 1.46 (s, 6H).

[0419] Fourth step: 2-(3-amino-4-bromothiophen-2-yl)propan-2-ol (56-5, 7 g) was dissolved in ethanol (70 mL), di-tert-butyl dicarbonate (32.34 g) was added, the reaction system was stirred at 50 °C for 16 hours, the reaction liquid was concentrated to obtain a crude product, and the crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 2: 1) to obtain compound 56-6 (3 g). LCMS (ESI) [M+H-t-Bu-H2O] + = 263.9; 1 H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 7.42 (s, 1H), 5.73 (s, 1H), 1.44 (s, 15H).

[0420] Fifth step: tert-butyl (4-bromo-2-(2-hydroxypropan-2-yl)thiophen-3-yl)carbamate (56-6, 3 g) was dissolved in methanol (30 mL), 1,1'-bis(diphenylphosphino)ferrocene 1,1'-bis(diphenylphosphino)ferrocene (494.61 mg), palladium acetate (200.31 mg), and triethylamine (4514.05 mg) were added in sequence, the reaction system was stirred at 70 °C for 16 hours under carbon monoxide atmosphere, the reaction liquid was concentrated to obtain a crude product, and the crude product was separated and purified by silica gel column (petroleum ether: tetrahydrofuran = 40:60) to obtain compound 56-7 (1.1 g). LCMS (ESI) [M+H-t-Bu+Na] + = 283.2; 1 H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 7.98 (s, 1H), 5.73 (s, 1H), 3.70 (s, 3H), 1.50-1.38 (m, 15H).

[0421] Sixth step: methyl 4-((tert-butoxycarbonyl)amino)-5-(2-hydroxypropan-2-yl)thiophene-3-carboxylate (56-7, 970 mg) was dissolved in methanol (10 mL) and water (5 mL), lithium hydroxide (368.3 mg) was added, the reaction system was stirred at 50 °C for 3 hours, methanol was removed by concentration, pH was adjusted to 5 with 1M hydrochloric acid, and extraction was performed with ethyl acetate (30 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to obtain compound 56-8 (800 mg). LCMS (ESI) [M+H-t-Bu-H2O] + = 228.1.

[0422] Step 7: 4-((tert-Butoxycarbonyl)amino)-5-(2-hydroxypropan-2-yl)thiophene-3- carboxylic acid (56-8, 800 mg) was dissolved in acetonitrile (8 mL), (S)-2-(4- (aminomethyl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester hydrochloride (1521.59 mg), TCFH (N,N,N',N'-tetramethyluronium hexafluorophosphate) (996.67 mg), N-methylimidazole (680.48 mg) were added successively, the reaction system was stirred at room temperature for 4 hours, the reaction solution was concentrated to obtain a crude product, and the crude product was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain compound 56-9 (650 mg). LCMS (ESI) [M+H-Boc-H2O] = 699.9. +

[0423] Step 8: (S)-2-(4-((4-((tert-butoxycarbonyl)amino)-5-(2-hydroxypropan-2-yl)thiophene-3- carboxamido)methyl)-2,6-dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (56-9, 400 mg) was dissolved in dichloromethane (4 mL), trifluoroacetic acid (2 mL) was added, the reaction system was stirred at room temperature for 1 hour, the pH was adjusted to 8 with saturated sodium bicarbonate, extracted with dichloromethane (15 mL*3), the combined organic phase was dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was separated and purified by silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain compound 56-10 (180 mg). LCMS (ESI) [M+H-H2O] = 700.2. +

[0424] Step 9: (S)-2-(4-((4-amino-5-(2-hydroxypropan-2-yl)thiophene-3-carboxamido)methyl)-2,6- dichlorobenzamido)-3-(3-(methylsulfonyl)phenyl)propanoic acid benzyl ester (56-10, 180 mg) was dissolved in tetrahydrofuran (5 mL), 1N sodium hydroxide (2.5 mL) was added, the reaction system was stirred at room temperature for 1 hour, the pH was adjusted to 7 with 1M hydrochloric acid, and concentrated to obtain a crude product, which was separated and purified by reverse phase system (water / ACN) to obtain target compound II-89 (10.69 mg). LCMS (ESI) [M+H-H2O] = 609.9; + 1 ​​​H NMR (400 MHz, DMSO-d6) δ 9.21 (s, 1H), 9.05 (s, 1H), 8.23 (s, 1H), 7.85 (s, 1H), 7.75 (d, J = 7.9 Hz, 1H), 7.66 (d, J = 7.7 Hz, 1H), 7.55 (t, J = 7.7 Hz, 1H), 7.36 - 7.32 (m, 2H), 4.73 - 4.61 (m, 1H), 4.42 (d, J = 5.8 Hz, 2H), 3.27 (s, 1H), 3.13 (s, 3H), 3.07 - 3.02 (m, 1H), 1.63 (s, 6H).

[0425] The following compounds were synthesized according to Reference Example 1-31:

[0426] Biological Experimental Example

[0427] Experimental Example 1: Complexation reaction of 4-hydroxynonenal

[0428] 1. Reagents and Instruments

[0429] The main reagents include 4-hydroxynonenal (Bid), Dulbecco's buffer.

[0430] The main instruments used in the experiment include a magnetic stirrer (Mingyinpu / 98-2) and a high-performance liquid chromatograph (Agilent / 1260).

[0431] 2. Experimental Method

[0432] 1) Preparation of reagents for complexation reaction of 4-hydroxynonenal:

[0433] a) Preparation of reference compound ADX 102 solution The compound ADX 102 was accurately weighed and dissolved in dimethyl sulfoxide to prepare a solution, and two groups were prepared.

[0434] b) Preparation of test compound solution

[0435] c) 4-Hydroxynonenal reaction solution preparation: Accurately weigh 4-hydroxynonenal (5.00 mg. 32.01 umol) into dimethyl sulfoxide (0.1 mL) to prepare a solution, and prepare 2 groups. Add ADX 102 solution and test compound solution into 8 ml single-mouth bottles, respectively, and add 1.25 ml of D-PBS (1.0X). Then add the prepared 4-hydroxynonenal solution, start stirring with a magnetic stirrer (30°C) and timing, and take samples for HPLC detection at 10 minutes, 100 minutes, 200 minutes and 300 minutes, respectively (use a pipette to take 25 uL of the upper emulsion layer and 50 uL of the aqueous phase, respectively, then dilute with 1 mL of methanol, and filter). The detection result is the percentage content of the complex product at 254 nm.

[0436] The compounds of the present application have the ability to capture toxic aldehydes, and the results of exemplary compounds are shown in Table 1.

[0437] Table 1: Complexation reaction of 4-hydroxynonenal with test compounds, ADX 102

[0438] Experimental Example 2: Malondialdehyde complexation reaction

[0439] 1 Reagents and instruments

[0440] The main reagents include malondialdehyde aqueous solution (Biodine). D-PBS (1X)

[0441] The main instruments used in the experiment include a magnetic stirrer (Meiyinpu / 98-2) and a liquid chromatograph-mass spectrometer (Waters).

[0442] 2 Experimental method

[0443] 2) Preparation of malondialdehyde reaction reagent:

[0444] a) Preparation of reference compound ADX 102 solution: accurately weigh compound ADX 102 (3.3 mg. 13.9 umol) into a 4 mL single-mouth bottle.

[0445] b) Preparation of test compound solution: accurately weigh the test compound (9.1 mg. 13.90 umol) into a 4 mL single-mouth bottle.

[0446] c) Preparation of reaction solution: add malondialdehyde aqueous solution (1.0 mL. 1.0 mg / ml) and D-PBS (0.5 mL, 1X) into the single-mouth bottle containing ADX 102 and the test compound. Start stirring with a magnetic stirrer (30°C) and timing, and take samples for LCMS detection at 10 minutes, 100 minutes, 200 minutes and 300 minutes, respectively (use a pipette to take 50 uL of the reaction solution under stirring, then dilute with 1 mL of methanol, and filter). The detection result is the percentage content of the complex product at 254 nm.

[0447] The compounds of the present application have the ability to trap toxic aldehydes, see Table 2 for results of exemplary compounds.

[0448] Table 2: Complexation of malondialdehyde with ADX102, test compounds

[0449] Experimental Example 3: Competition assay testing of antagonists in LFA-1 / ICAM-1 and LFA-1 / small molecule ELISA

[0450] 1 Reagents and Instruments

[0451] 2 Experimental Methods:

[0452] Human recombinant ICAM-1 / CD54 protein was prepared in TBS buffer to a final concentration of 1 ug / mL, 50 uL of the solution was transferred to a 96-well plate and incubated at 4°C overnight. The plate was washed 3 times with buffer, then 150 uL of blocking solution was added and incubated at 37°C for 1 h. The plate was washed 3 times with buffer, 1 ug / mL of human integrin aL / beta2 dimer protein was prepared in buffer containing 0.1% BSA, 50 uL of the integrin protein was transferred to the 96-well plate, 1 uL of different concentrations of compounds or DMSO was added, and incubated at room temperature for 2 h. 1 ug / mL of human integrin beta2 / CD18 biotinylated antibody was prepared in buffer containing 0.1% BSA, the plate was washed 3 times with buffer, 50 uL of the antibody was added and incubated at room temperature for 1 h. The plate was washed 3 times with buffer, 50 uL of Streptavidin-HRP was added and incubated at room temperature for 20 min. The plate was washed 3 times with buffer, 50 uL of TMB substrate was added and incubated at room temperature for 20 min. Finally, 25 uL of stop buffer was added, and the OD value of the plate was read at 450 nm on a microplate reader. The IC50value was calculated using GraphPad Prism 5 software. The compounds of the present application can effectively inhibit the binding of LFA-1 to ICAM-1. See Table 3 below for results of exemplary compounds. 50

[0453] Table 3: Results of compound competition binding assay Note: A: less than or equal to 100 nM.

[0454] Experimental Example 4: Jurkat cell adhesion inhibition assay testing

[0455] 1 Reagents and Instruments

[0456] 2 Experimental Methods:

[0457] ​R&D, Cat# 720-IC) was prepared at 5 μg / mL in Tris-Buffered Saline buffer (Boston BioProducts, Cat# BM-300) and 50 μL of the solution was transferred to a 96-well plate (Corning, Cat# 3690) for overnight coating at 4°C. The plate was washed 3 times with Tris-Buffered Saline buffer, and 150 μL of 0.1% BSA blocking solution (BSA dissolved in Tris-Buffered Saline buffer) was added for 1 h blocking at 37°C. The plate was washed 3 times with Tris-Buffered Saline buffer, and Jurkat cells (ATCC, Cat# TIB 152) were resuspended in Tris-Buffered Saline buffer and 200,000 cells / 50 μL were transferred to the 96-well plate, and 1 μL of different concentrations of compounds or dimethyl sulfoxide (Sigma, Cat# 34943) was added for 2 h incubation at room temperature. The liquid in the 96-well plate was removed, and the plate was washed 2 times with Tris-Buffered Saline buffer, and 50 μL of substrate (4-nitrophenyl-N-acetyl-β-D-glucosamine) (TCI, Cat# N0866) was added for 2 h incubation at 37°C. 90 μL of stop buffer was added, and the OD value of the plate was read at 405 nm on a microplate reader FlexStation 3 (Molecular Devices). The IC50values were calculated using GraphPad Prism 5 software. The compounds of the present application have good inhibitory effect on the adhesion ability of Jurkat cells, and the results of exemplary compounds are shown in Table 4. 50 Table 4: Results of compound cell adhesion experiment

[0458] Table 4: Results of compound cell adhesion experiment

[0459] IC50values of compound cell adhesion experiment inhibition: A≤20 nM, 20 nM < B≤50 nM, 50 nM < C≤100 nM. 50 Table 4: Results of compound cell adhesion experiment

[0460] Experimental Example 5: Compound rabbit eye tissue distribution study test

[0461] 1 Animal information: 2-3 kg female New Zealand white rabbits were purchased from Jiashan Jinrui Rabbit Industry Professional Cooperative.

[0462] 2 Test compound preparation: Each 17 mg of test compound was weighed into 0.9 mL of sterile normal saline, 7.30 mg of NaOH was added, vortexed to mix, then saturated aqueous solution of sodium phosphate monobasic was added to adjust the pH to between 6.75 and 7.25, then sterile normal saline was added to bring the total volume to 1 mL. This resulted in a solution containing the test compound, each compound at a concentration of 17 mg / mL, which was prepared immediately before use.

[0463] 3 Sample collection

[0464] 6 After completion of the ocular administration of the test compound (50 μL per eye) in both eyes of each animal, tissue and blood samples were collected at 0.5 and 2.0 hours. Approximately 500 μL of blood sample was collected into an anticoagulant tube containing EDTA-K2, and centrifuged to obtain plasma within 60 minutes. The whole blood sample was 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 at each of 0.5 and 2 hours), and ocular tissues were collected from both eyes: cornea, bulbar conjunctiva, and palpebral conjunctiva. The ocular tissues were rinsed with normal saline, surface moisture was absorbed with filter paper, and the tissues were weighed and stored. All samples were weighed and stored separately. A total of 12 corneas, 12 bulbar conjunctivas, and 12 palpebral conjunctivas were collected. All collected plasma samples were stored on dry ice or in a freezer until analysis.

[0465] 4 Sample processing and detection method

[0466] Plasma standard curve, quality control, and sample (blank and test) extraction process: 70.0 μL of internal standard working solution was added to each of the 96-well plates that had been prepared with 7.0 μL of the prepared blank, standard curve, quality control, and test sample, respectively. The 96-well plates were mixed on a vortex mixer for 10 minutes (temperature set to room temperature, and rotation speed set to 500 rpm), centrifuged for 10 minutes (temperature set to 4°C, and centrifugal force set to 4000 rpm), and 50.0 μL of supernatant was transferred to a 96-well plate that had been prepared with 50.0 μL of ultrapure water. The 96-well plate was mixed on a vortex mixer for 10 minutes (temperature set to room temperature, and rotation speed set to 500 rpm).

[0467] The blank ocular tissues of the rabbit, cornea, bulbar conjunctiva, and palpebral conjunctiva were homogenized with 50% acetonitrile solution at a weight-to-volume ratio of 1:5 (w / v) to obtain ocular tissue homogenate.

[0468] Eye tissue homogenate solution standard curve, quality control and sample (blank and to be tested) extraction process: 30.0 μL of prepared blank, standard curve and quality control sample were added to a 96-well plate, respectively, 300.0 μL of internal standard working solution was added, the 96-well plate was placed in a homogenizer for 10 minutes (temperature set to room temperature, rotation speed set to 500 rpm), centrifuged for 10 minutes (temperature set to 4°C, centrifugal force set to 4000 rpm), 100.0 μL of supernatant was taken to a 96-well plate pre-added with 100.0 μL of ultrapure water, the 96-well plate was placed in a homogenizer for 10 minutes (temperature set to room temperature, rotation speed set to 500 rpm).

[0469] 5 Test results:

[0470] Table Five

[0471] The test results show that II-23 and II-44 have high drug concentrations in the cornea, bulbar conjunctiva and eyelid conjunctiva after eye drop administration in rabbits, and low drug concentrations in the plasma.

[0472] Experimental Example 6: Scopolamine-induced dry eye model test in mice

[0473] 1 Animal information: 8-10 week C57BL / 6J male mice, purchased from Shanghai Jihui Experimental Animal Breeding Co., Ltd.

[0474] 2 Preparation of test compounds: weigh an appropriate amount of test compound 5 g into 90 mL of sterile normal saline, add 0.7 g of NaOH, stir to obtain a transparent solution; add NaH2PO4(sodium phosphate) saturated aqueous solution to the obtained solution until the pH of the solution is between 6.75-7.25 (use precise pH test paper to measure pH value or pH meter), add sterile normal saline to the obtained aqueous solution until the total volume reaches 100.0 mL. The obtained solution is divided into sterile EP tubes according to the amount used per day and stored at 2-8°C in the dark. The vehicle is the system without the compound. The 1% (mass fraction) solution is diluted to obtain a 5% (mass fraction) solution.

[0475] 3 Experimental reagents and instruments

[0476] Scopolamine hydrobromide (Mydriacyl), Restasis eye emulsion (Allergan Sales, LLC), fluorescein sodium injection (Alcon Laboratoryes. Inc); handheld slit lamp (SL-17, Kowa)

[0477] 4 Model creation environment:

[0478] Mice will be housed in clear resin plastic cages (390 mm x 180 mm x 130 mm) in the animal room, 2-5 mice per cage. The cage bedding will be autoclaved corn cob bedding (Dachang Hui Autonomous County Chenfu Eden Bedding Processing Factory), changed every 7-10 days. The room number where the animals are housed will be recorded in the experimental records throughout the experiment. The animal room will be equipped with high efficiency particulate air filters, with 15-25 air changes per hour. The temperature will be maintained between 20-26 °C (68-79 °F), with a relative humidity of 30-70%. Temperature and humidity will be continuously observed and recorded. The lighting conditions will be 12 hours of fluorescent light (08:00-20:00) and 12 hours of darkness per day.

[0479] 5. Experimental design

[0480] 5.1. Modeling

[0481] Before modeling, detailed eye observation will be performed using a slit lamp, and animals with no abnormalities in both eyes will be selected for the study, of which 6 animals will be used as the G1 normal control group, and the remaining animals will be used for modeling.

[0482] After grouping on Day-1, all animals in the study except for G1 will be housed in a dry box to control the environmental relative humidity to be less than 40%, and G2-G8 animals will be subcutaneously injected with 0.25% Scopolamine Hydrobromide (SCO) for modeling, and G1 animals will be subcutaneously injected with Normal Saline (NS) as the normal control group, with a dosage of 0.2 mL / time, 3 times per day (time points: 9:00, 13:00, 17:00, ±1 h), for 14 days.

[0483] 5.2. Grouping and administration

[0484] Before administration, the animals will be relatively evenly grouped according to the amount of tear secretion.

[0485] Group 1-Group 8: From Day 1 to Day 13, eye drops will be administered bid, 15 μL / eye / time (only once in the morning on Day 14). See Table 6 for details of grouping and administration.

[0486] Table 6

[0487] 5.3. Detection indicators

[0488] Schirmer I test (SIT)

[0489] Schirmer I test was performed to evaluate the tear secretion of all animals in the study. The mice were fixed and a piece of phenol red soaked cotton thread was inserted into the temporal conjunctival sac of the lower eyelid with microforceps. The length of the thread was measured after 30 seconds and the change in color was recorded with a vernier caliper. If the thread fell out, a new one was inserted after 5 minutes.

[0490] CFS score

[0491] CFS score was performed to evaluate the ocular surface damage of all animals in the study. The mice were fixed and 20 μL of 0.5% sodium fluorescein solution was instilled into the upper conjunctival sac. The sodium fluorescein was evenly distributed on the cornea with the aid of blinking. The cornea was then washed with saline. The cornea was observed under a slit lamp with cobalt blue light and the fluorescence staining was evaluated according to a 16-point scale. The center of the cornea was taken as the origin and divided into four quadrants. Each quadrant was scored according to the standard shown in Table 7. Each eye of each animal scored 0-16 points.

[0492] Table 7. CFS score standard

[0493] 6 Statistical analysis

[0494] The data were expressed as mean ± standard error. Statistical analysis was performed using Graphpad Prism, SPSS or Sigmaplot software. The specific data were presented in the form of charts. P<0.05 was considered statistically significant.

[0495] As shown in Figures 1 and 2, in the scopolamine-induced dry eye model test, the tear secretion and corneal fluorescence staining were significantly improved at the concentrations of 1% and 5% of II-44.

[0496] In Figure 1, *p<0.05, ***p<0.001: Day 14, G1 vs G2, G5 vs G2, G6 vs G2 had significant differences (Unpaired t test); *p<0.05, **p<0.01: Day 14, G5 vs G3, G6 vs G3 had significant differences (Unpaired t test).

[0497] In Figure 2, *p<0.05, **p<0.01, ***p<0.001: Day 14, G1 vs G2, G4 vs G2, G5 vs G2 had significant differences (Unpaired t test); *p<0.05: Day 14, G5 vs G3 had significant differences (Unpaired t test).

[0498] Experimental Example 7: Melanin binding test

[0499] Experimental procedure:

[0500] 1. Pre-warm DPBS buffer.

[0501] 2. Preparation of melanin dosing solution

[0502] Melanin was stored in a refrigerator at -20°C. Before use, melanin was dissolved in DPBS at a concentration of 1 mg / mL and sonicated for 5 minutes at 37°C to obtain a suspension.

[0503] 3. Preparation of test and positive control working solutions (0.2 mM)

[0504] Add 2 μL of 10 mM stock solution to 98 μL of DMSO.

[0505] 4. Preparation of test and positive control dosing solutions

[0506] Add 1194 μL of melanin dosing solution to a 96-well plate, then add 6 μL of test and positive control working solutions (0.2 mM) with a final test concentration of 1 μM.

[0507] 5. T0 preparation

[0508] Add 50 μL of test and positive control dosing solutions to a 96-well sample preparation plate as T0 samples, then add 300 μL of internal standard (IS) containing termination solution, vortex the samples at 600 rpm for 10 minutes, and store the sample plate in a refrigerator (-20°C).

[0509] 6. Centrifugation step

[0510] 6.1 Take 300 μL of dosing solution to a 96-well plate, incubate the samples at 37°C at 750 rpm for 60 minutes, then centrifuge at 1600 x g for 10 minutes.

[0511] 6.2 Continue incubation of the remaining dosing solution at 37°C until the end of centrifugation to obtain T samples.

[0512] 7. Preparation of samples after centrifugation

[0513] 7.1 F samples: After centrifugation (step 6.1), take 50 μL of supernatant and add it to a new sample preparation plate.

[0514] 7.2 T samples: Transfer 50 μL of samples after the end of incubation (step 6.2.) to a new sample preparation plate

[0515] 7.3 Then add 300 μL of stop solution containing internal standard (IS). Vortex all samples at 600 rpm for 10 minutes, then centrifuge at 6000 rpm for 15 minutes.

[0516] 7.4 Take 100 μL of supernatant into a 96-well sample plate containing 100 μL of ultrapure water for LC / MS analysis.

[0517] 8 Data processing:

[0518] % Fu = 100 x F / T

[0519] % Bound = 100 x (1 - F / T)

[0520] % Recovery = 100 x T / T0

[0521] The experimental results show that the compound of the application has weak binding to melanin and good compound safety.

Claims

A compound represented by the following formula (A), a stereoisomer, a tautomer, or a mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound: wherein L is or optionally substituted: C 1-6 alkylene, C 2- 6alkenylene, C 2-6 alkynylene, C 1-6 alkyleneoxy, C 1-6 alkylthio; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from deuterium, halogen, oxo, -OH, -SH, -NH2, C 1-4 alkyl; * indicates the point of attachment of ring A; n is 1, 2, or 3; Z1, Z2, Z3and Z4are each independently selected from N or CR Z ; R Z each independently is hydrogen, deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-6 alkyl, C 3-6 cycloalkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2; Ring A is 5-12 membered heteroaryl or C 6-12 aryl; Ring B is a 5-12 membered heteroaryl or C 6-12 aryl; R A each independently deuterium, halogen, -CN, -OH, -NH2, or is optionally substituted: C 1-6 alkyl, C 2-6 alkenyl, C 3-8 cycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2, C 1-4 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, C 3-6 cycloalkyl; R B each independently deuterium, halogen, -CN, -NH2, methylsulfonyl, ethylsulfonyl, -S(O)2NH2, -P(O)(CH3)2, -P(O)2CH3, -S(O)(NH)CH3, or is optionally substituted: C 1-6 alkyl, C 3-8 cycloalkyl, C 1-6 alkoxy, C 1-6 alkylthio; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, -OH, -SH, -NH2, C 1-4 alkyl. Rc is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl; Rd is selected from hydrogen, deuterium, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl; R f1 and R f2 are each independently selected from hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-6 alkyl, C 1-6 haloalkyl, C 3-8 cycloalkyl, C1-6alkoxy, C 1-6 alkylthio; or R f1 and R f2 form together with the C atom to which they are attached a C 3-8 cycloalkyl; R1and R2are each independently selected from hydrogen, halogen, C 1-6 alkyl, C 1-6 haloalkyl; a is selected from 2, 3, 4, 5, or 6; b is selected from 1, 2, 3, 4, 5, or 6; Unless otherwise indicated, the heteroatoms in the above heteroaryl, heterocyclyl groups are independently selected from O, N, or S, and the number of heteroatoms is 1, 2, 3, or 4; provided that, two of R A -NH2and at the time, -NH2and adjacent. The compound of claim 1, stereoisomers, tautomers, or mixtures thereof, pharmaceutically acceptable salts of the compound, wherein: L is or optionally substituted: C 1-3 alkylene, C 2-6 alkynylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from deuterium, halogen, oxo, -OH, -SH, -NH2, C 1-4 alkyl; * indicates the point of attachment of ring A. Z1, Z2, Z3and Z4are each independently selected from N or CR Z ; R Z is hydrogen, deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-3 alkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -OH, -SH, -NH2; Rc is selected from hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl; R f1 and R f2 are each independently selected from hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-3 alkyl, C 1-3 haloalkyl, C 3-6 cycloalkyl; or R f1 and R f2 form together with the C atom to which they are attached a C 3-6 cycloalkyl; Alternatively, L is or optionally substituted C 1-3 alkylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from deuterium, halogen, -OH, -SH, -NH2; * indicates the point of attachment to ring A; one, two, or three of Z1, Z2, Z3, and Z4are N, and the rest are CH; Rc is selected from hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl; R f1 and R f2 are each independently selected from the group consisting of hydrogen, deuterium, halogen, C 1-3 alkyl; or R f1 and R f2 form together with the C atom to which they are attached a C 3-6 cycloalkyl; Alternatively, L is * indicates the ring A attachment point; Rc is selected from hydrogen, deuterium, C 1-3 alkyl, C 3-6 cycloalkyl; R f1 and R f2 are each independently selected from the group consisting of hydrogen, deuterium, halogen, -CN, -OH, -NH2, C 1-3 alkyl; or R f1 and R f2 form together with the C atom to which they are attached a C 3-6 cycloalkyl; Alternatively, L is * denotes the ring A attachment point; wherein Rc is hydrogen, methyl, ethyl or cyclopropyl; R f1 and R f2 are each independently selected from hydrogen, deuterium and methyl; or R f1 and R f2 together with the C atom to which they are attached form a cyclopropyl group; Alternatively, L is * indicates the ring A attachment point; or L is -CONH(CH2)2-; Alternatively, L is or optionally substituted C 1-3 alkylene; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from deuterium, halogen, oxo, -OH, -SH, -NH2, C 1-4 substituted with one or more substituents selected from deuterium, halogen, oxo, -OH, -SH, -NH2, C * indicates the point of attachment of ring A. one, two or three of Z1, Z2, Z3and Z4are N, and the others are CR Z (or the others are CH); R Z is hydrogen, deuterium, halogen, -CN, -OH, -NH2or is optionally substituted: C 1-3 alkyl; wherein optionally substituted means unsubstituted or substituted with one or more substituents selected from the group consisting of deuterium, halogen, -NH2; Alternatively, L is * indicates the ring A attachment point; or L is -CH2-. A compound according to claim 1 or 2, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt thereof, wherein: ring A is 5-10 membered heteroaryl or phenyl; or ring A is pyridyl, quinolinyl, or phenyl; or ring A is phenyl. A compound according to any one of claims 1 to 3, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: a is 2, 3 or 4, wherein two R A are -NH2and -OH, respectively adjacent. or a is 2 and both R A are located at the ortho position, both R A are -NH2 and Alternatively, a is 3, and wherein two R A are located in the ortho position, two R A are each -NH2 and another R A selected from deuterium, halogen, -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 wherein two R A groups are located in the ortho position, two R A groups are -NH2 and Another R A selected from halogen (e.g., F) or -OH. A compound according to any one of claims 1 to 4, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: For or For or For A compound according to any one of claims 1 to 5, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: ring B is 6-10 membered heteroaryl or phenyl; preferably, ring B is phenyl. A compound according to any one of claims 1 to 6, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: R B -NH2, methylsulfonyl, -S(O)2NH2, -P(O)(CH3)2, -P(O)2CH3, -S(O)(NH)CH3or or b is 1 and R B is methanesulfonyl, -S(O)2NH2, -P(O)(CH3)2, -S(O)(NH)CH3; or b is 1 and R B is methanesulfonyl. A compound according to any one of claims 1 to 7, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: For or For or For A compound according to any one of claims 1 to 8, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: Rd is hydrogen. A compound according to any one of claims 1 to 9, a stereoisomer, a tautomer or a mixture thereof, a pharmaceutically acceptable salt of said compound, wherein: R1and R2are each independently selected from hydrogen, halogen; or R1and R2are each independently selected from hydrogen, halogen, and wherein at least one is halogen; or R1and R2are each independently selected from hydrogen, Cl; and wherein at least one is Cl; or R1and R2are each halogen; or R1and R2are each Cl. A compound of Formula (I-a), (I-b), (I-c), (I-d), (IV-a), (IV), (IV’), or (V), a stereoisomer, tautomer, or mixture thereof of the compound, or a pharmaceutically acceptable salt of the compound: wherein, ring A, ring B, R1, R2, R A , a, R B , b, R f1 , R f2 , R c , R d as described in any one of claims 1 to 10 for a compound of formula (A); wherein ring A, ring B, R1, R2, R A , a, R B , b, R f1 , R f2 , R c , R d as described in any one of claims 1 to 10 for the use of a compound of formula (A); wherein ring A, ring B, R1, R2, R A , a, R B , b, R d as described in any one of claims 1 to 10 for compound of formula (A); wherein ring A, ring B, R1, R2, R A , a, R B , b, R d as described in any one of claims 1 to 10 for the compound of formula (A); wherein ring A, ring B, R1, R2, R A , a, R B , b, Rc, R f1 , and R f2 are as described in claim 1-10 for compound of formula (A); wherein ring A, ring B, R1, R2, R A , a, R B , b are as described for the compound of formula (A) according to any one of claims 1 to 10; wherein ring A, ring B, R f1 , R f2 , R A , a, R B , b, R1, R2 are as described for the compound of formula (A) according to any one of claims 1 to 10; wherein ring A, R1, R2, R A , a compound of formula (A) as described in any one of claims 1-10. a compound, a stereoisomer, a tautomer of the compound, or a mixture thereof, or a pharmaceutically acceptable salt of the compound: a compound, a stereoisomer, a tautomer of the compound, or a mixture thereof, or a pharmaceutically acceptable salt of the compound: A pharmaceutical composition comprising a compound of any one of claims 1-13, stereoisomers, tautomers, or mixtures thereof, pharmaceutically acceptable salts of the compound, optionally further comprising a pharmaceutically acceptable carrier. Use of a compound of any one of claims 1-13, stereoisomers, tautomers, or mixtures thereof, pharmaceutically acceptable salts of the compound, or a pharmaceutical composition of claim 14, for the manufacture of a medicament for the treatment and / or prevention of a LFA-1 mediated and / or an aldehyde toxicity involved pathogenesis disease; optionally, the LFA-1 mediated and / or an aldehyde toxicity involved pathogenesis disease is an ocular disease; optionally, the ocular disease is a chronic ocular surface disease; optionally, the ocular disease is dry eye.

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