Interleukin-17A / F inhibitor as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of chemical drugs, provides an interleukin-17A / F inhibitor as well as a preparation method and application thereof, and particularly relates to a compound as shown in a general formula (I), or a racemate, an isomer or medicinal salt thereof, which is used for treating diseases such as psoriasis and arthritis as the IL-17A / F inhibitor.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical drugs and relates to a class of interleukin-17A / F small molecule inhibitors, and specifically relates to an interleukin-17A / F inhibitor and a preparation method and application thereof. Background Art

[0002] The IL-17 family consists of six cytokines (IL-17A to IL-17F). Interleukin-17A (IL-17A) is an established proinflammatory cytokine that is involved in the induction of IL-6, IL-8, G-CSF, TNF-a, IL-ip, PGE2 and IFN-y, as well as many chemokines and other effectors. IL-17A can form homodimers or heterodimers with its family member IL-17F and can bind to the IL-17 receptors IL-17RA and IL-17RC to mediate signal transduction. IL-17A is the main pathological cytokine expressed by Th 17 cells, involved in the pathology of inflammation and autoimmunity, and also involved in CD8+ T cells, y6 cells, NK cells, NKT cells, macrophages and dendritic cells. IL-17A is a well-recognized proinflammatory cytokine that plays a key role in chronic inflammation and is a major driver of tissue damage. IL-17A induces normal immune and inflammatory responses to pathogens but can also contribute to chronic autoimmune diseases, including psoriasis, spondyloarthritis, rheumatoid arthritis, and multiple sclerosis.

[0003] Although many patents for IL-17A small molecule inhibitors have been published, for example, patent publication numbers WO2020182666, WO2023283453, WO2023275301, and WO2023025783, there is currently no effective oral treatment for moderate to severe psoriasis. Therefore, there is a need to develop effective small molecule IL-17A inhibitors to treat inflammatory diseases and other diseases. Summary of the Invention

[0004] In view of the problems existing in the prior art, the present application provides an interleukin-17A / F inhibitor and its preparation method and application. As an IL-17A / F inhibitor, the compound can inhibit the production of cytokines IL-17A / F, thereby regulating inflammatory responses and other related diseases.

[0005] In a first aspect, the present application provides a compound represented by general formula (I), or a racemate, an isomer, or a pharmaceutically acceptable salt thereof:

[0006]

[0007] In a second aspect, the present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of any one of the above-mentioned compounds or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0008] In a third aspect, the present invention further provides a use of a therapeutically effective amount of the above-mentioned compound or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating a disease, wherein the disease is an IL-17A / F-related disease, specifically, the disease is selected from psoriasis and arthritis.

[0009] Specifically, the present invention is achieved through the following technical solutions:

[0010] In a first aspect, the present invention provides a compound represented by formula (I) or its racemate, or its isomer, or its pharmaceutically acceptable salt,

[0011]

[0012] Wherein, n is 0, 1, 2, 3 or 4;

[0013] Ring A is selected from R 1 Substituted heterocycloalkyl, aryl, heteroaryl, the R 1 is selected from alkyl, halogen, cycloalkyl, haloalkyl, haloalkoxy, halocycloalkyl, -O-cycloalkyl;

[0014] R 2 is selected from substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -CH(R 2’ )(R 2” ), the substituent is selected from alkyl, halogen, haloalkyl, haloalkenyl, the R 2’ and R 2” are independently selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl;

[0015] R 3 is selected from hydrogen, halogen, alkyl; R 4 is selected from hydrogen, halogen, alkyl, cycloalkyl;

[0016] X 1 、X 2 solely selected from hydrogen, Or, X 1 、X 2 Cyclize together to form a substituted or unsubstituted heterocycloalkyl or heteroaryl, wherein the substitution is selected from alkyl, halogen;

[0017] R 5a 、R 5b independently selected from alkyl, alkenyl, alkynyl, nitrile, alkylnitrile, haloalkyl, haloalkenyl, substituted or unsubstituted -(CH2) k -O-alkyl, -(CH2)p -O-cycloalkyl, -(CH2) q B1B2-O-alkyl, cycloalkyl, heterocycloalkyl, alkylheterocycloalkyl, halogenated heterocycloalkyl, aryl, heteroaryl, the substituent is selected from alkyl, halogen, alkoxy, B1 and B2 are independently selected from alkyl and halogen, or B1 and B2 are cyclized together to form a substituted or unsubstituted cycloalkyl or heterocycloalkyl, the substituent is selected from alkyl and halogen;

[0018] R 6 and R 7 independently selected from alkyl, alkoxy, cycloalkyl, or R 6 and R 7 Cyclize together to form a substituted or unsubstituted heterocycloalkyl, wherein the substitution is selected from alkyl, alkyl alcohol, alkoxy, alkoxyamide, halogen, cycloalkyl, -(CH2) k -O-alkyl, haloalkyl;

[0019] k, m, p, q, y are independently selected from 1, 2, 3, 4, 5 or 6;

[0020] Among them, when R 5a Selected from -CH2-O-alkyl or alkyl, said R 6 and R 7 Cyclize together When the A ring is not

[0021] When R 5a When selected from alkyl, the R 6 and R 7 Cyclize together to form a substituted heterocycloalkyl, wherein the substitution is selected from alkoxyamide;

[0022] When R 5a is selected from alkyl groups, wherein R 6 and R 7 Cyclize together When the R 2 for When R 5a When selected from unsubstituted heteroaryl, the R 6 and R 7 Cyclize together When R 5a Selected from When the R 2 Not for When R 2 Selected from When n is not 0 or 1, or R 6 and R 7 Cyclize together to form a heterocycloalkyl group substituted by halogen.

[0023] As a preferred technical solution of the present invention, the alkyl group is selected from C 1-6 The alkyl group, the C 1-6 The alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, and 1-ethylbutyl;

[0024] The alkenyl group is selected from C 2-6 Alkenyl, C 2-6The alkenyl group is selected from the group consisting of vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 3-methyl-1-butenyl, 1-methyl-2-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl. , 3-methyl-3-butenyl, 1,1-dimethyl-2-propenyl, 1,2-dimethyl-1-propenyl, 1,2-dimethyl-2-propenyl, 1-ethyl-1-propenyl, 1-ethyl-2-propenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 3-methyl-1-pentenyl, 4-methyl-1-pentenyl, 1-methyl-2-pentenyl, 2-methyl-2-pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl, 1-methyl-3-pentenyl 1-methyl-4-pentenyl, 2-methyl-4-pentenyl, 3-methyl-4-pentenyl, 4-methyl-4-pentenyl, 1,1-dimethyl-2-butenyl, 1,1-dimethyl-3-butenyl, 1,2-dimethyl-1-butenyl, 1,2-dimethyl-2-butenyl, 1,2-dimethyl-3-butenyl, 1,3-dimethyl-1-butenyl, 1,3-dimethyl-2-butenyl, 1,3-dimethyl-3-butenyl, 2,2-dimethyl-3-butenyl 1-ethyl-2-methyl-1-propenyl, 1-ethyl-2-methyl-2-propenyl, 1-ethyl-3-butenyl, 2-ethyl-1-butenyl, 2-ethyl-2-butenyl, 2-ethyl-3-butenyl, 1,1,2-trimethyl-2-propenyl, 1-ethyl-1-methyl-2-propenyl, 1-ethyl-2-methyl-1-propenyl, and 1-ethyl-2-methyl-2-propenyl;

[0025] The alkynyl group is selected from C 2-6 The alkynyl group, the C 2-6 The alkynyl group is selected from ethynyl, propynyl, 2-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl;

[0026] The alkyl nitrile is selected from acetonitrile, propionitrile, butyronitrile, isobutyronitrile, valeronitrile, isovaleronitrile, and hexanenitrile;

[0027] The aryl group is selected from a five-membered or six-membered aryl group; the heteroaryl group refers to a group in which at least one carbon atom on the aryl group is substituted by a heteroatom.

[0028] The heteroaryl group is selected from a 5-membered or 6-membered heteroaryl group, and the 5-membered or 6-membered heteroaryl group is selected from

[0029] As a preferred technical solution of the present invention, the cycloalkyl group is selected from C 3-12 Cycloalkane, C 3-12 The cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclic alkyl, bridged cycloalkyl, and spirocyclic alkyl; the bicyclic alkyl means that two rings share two carbon atoms, the bridged cycloalkyl means that two rings share two or more carbon atoms, and the spirocyclic alkyl means that two cycloalkyls share one carbon atom; the heterocycloalkyl means that at least one carbon atom on the cycloalkyl is substituted by a heteroatom; the alkylcycloalkyl means that at least one hydrogen atom on the cycloalkyl is substituted by an alkyl group; the alkylheterocycloalkyl means that at least one hydrogen atom on the heterocycloalkyl is substituted by an alkyl group; the halogenated heterocycloalkyl means that at least one hydrogen atom on the heterocycloalkyl is substituted by a halogen.

[0030] As a preferred technical solution of the present invention, the spirocycloalkyl group is selected from

[0031] The bridged cycloalkyl group is selected from: The bicyclic alkyl group is selected from:

[0032] As a preferred technical solution of the present invention, the halogen is selected from fluorine, chlorine, bromine and iodine; and a haloalkyl group refers to an alkyl group in which at least one hydrogen atom is replaced by a halogen.

[0033] As a preferred technical solution of the present invention, the heteroatom is selected from nitrogen, oxygen, and sulfur, and the heteroatom is one or more.

[0034] Specifically, the heterocycloalkyl group is selected from

[0035] As a preferred technical solution of the present invention, ring A is selected from

[0036] The R 1 The substituted A ring is selected from

[0037] R 2 Selected from cycloheptyl,

[0038] n is 0 or 1, m, y are 1, k is 1 or 2, p, q are 1; R3 is selected from hydrogen, methyl or fluorine;

[0039] X 1 、X 2 solely selected from hydrogen, or X 1 、X 2 Cyclize together

[0040] R 5a Selected from ethyl, vinyl, ethynyl, propadiene, nitrile, acetonitrile,

[0041] R 6 and R 7 independently selected from methyl, methoxy, methoxymethyl, or R 6 and R 7 Cyclize together

[0042] And when R 5a Selected from When R 6 and R 7 Not cyclized together When R 5a When selected from ethyl, R 6 and R 7 Cyclize together When R 5a When selected from ethyl, the R 6 and R 7 Cyclize together When the R 2 for When R 5a Selected from When the R 6 and R 7 Cyclize together

[0043] As a preferred technical solution of the present invention, the compound, or its racemate, or its isomer, or its pharmaceutically acceptable salt, is selected from the compound represented by formula (Ia), or its racemate, or its isomer, or its pharmaceutically acceptable salt:

[0044] Where n is 0 or 1, R 2 Selected from cycloheptyl, A ring, R 3 、X 1 、X 2 、R 6 、R7 As defined above.

[0045] As a preferred technical solution of the present invention, the compound, or its racemate, or its isomer, or its pharmaceutically acceptable salt is selected from the compounds shown in Table 1.

[0046] As a preferred technical solution of the present invention, the pharmaceutically acceptable salt refers to a compound, or an isomer thereof, or a racemate thereof, or a pharmaceutically acceptable salt thereof prepared with a pharmaceutically acceptable acid or base.

[0047] In a second aspect, the present invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the compound as described in any one of the above items, or its racemate, or its isomer, or its pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

[0048] In a third aspect, the present invention also provides a use of the compound, or its racemate, or its isomer, or its pharmaceutically acceptable salt in the preparation of a drug for treating a disease, wherein the disease is an IL-17A-related disease, specifically selected from psoriasis, arthritis and the like.

[0049] For the sake of clarity, general terms used in the description of the compounds are defined herein.

[0050] Unless otherwise indicated, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered as undefined or unclear in the absence of a specific definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient. The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues within the scope of sound medical judgment without excessive toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0051] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, which is prepared by reacting the compound having specified substituents discovered in the present invention with a pharmaceutically acceptable acid or base.

[0052] In addition to the form of salts, the compounds provided by the present invention also exist in prodrug form. The prodrugs of the compounds described herein easily undergo chemical changes under physiological conditions to be converted into the compounds of the present invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an in vivo environment.

[0053] Certain compounds of the present invention may exist in unsolvated forms as well as solvated forms, including hydrates. In general, the solvated forms are equivalent to the unsolvated forms and are encompassed within the scope of the present invention.

[0054] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, atropisomers, and racemic and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0055] Optically active (R)- and (S)-isomers, as well as D and L isomers, atropisomers, etc., can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are resolved by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished by using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate formation from an amine).

[0056] The atoms of the molecules of the compounds of the present invention are isotopes, and isotope derivatization can generally extend half-life, reduce clearance, stabilize metabolism, and increase in vivo activity. In addition, an embodiment is included in which at least one atom is replaced by an atom having the same atomic number (number of protons) and a different mass number (protons and neutrons). Examples of isotopes included in the compounds of the present invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, chlorine atoms, which respectively include 2 H. 3 H. 13 C. 14 C. 15 N. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 36 In particular, radioactive isotopes that emit radiation as they decay, such as 3 H or 14C can be used for local anatomy testing of pharmaceutical preparations or compounds in vivo. Stable isotopes neither decay nor change with their amount nor are they radioactive, so they can be used safely. When the atoms constituting the molecules of the compounds of the present invention are isotopes, the isotopes can be converted according to general methods by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0057] The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, the compounds may be labeled with radioactive isotopes, such as deuterium ( 2 H), iodine-125( 125 I) or C-14( 14 C) All isotopic variations of the compounds of the present invention, whether radioactive or not, are encompassed within the scope of the present invention.

[0058] Furthermore, the compounds of the present invention may have one or more hydrogen atoms replaced by deuterium isotopes ( 2 After deuteration, the compounds of the present invention have the effects of extending half-life, reducing clearance rate, stabilizing metabolism and improving in vivo activity.

[0059] The preparation method of the isotopic derivative generally includes a phase transfer catalytic method. For example, a preferred deuteration method uses a phase transfer catalyst (e.g., a tetraalkylammonium salt, NBu4HSO4). The use of a phase transfer catalyst to exchange the methylene protons of the diphenylmethane compound results in a higher deuterium incorporation than reduction with a deuterated silane (e.g., triethyldeuterated monosilane) in the presence of an acid (e.g., methanesulfonic acid) or with a Lewis acid such as aluminum trichloride using sodium deuterated borate.

[0060] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and has no toxic side effects on the host or patient. Representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, viscosity increasing agents, transdermal enhancers, etc. Their preparations are well known to those skilled in the art of cosmetics or topical medicine. For additional information about carriers, reference can be made to Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the contents of which are incorporated herein by reference.

[0061] The term "excipient" generally refers to a carrier, diluent and / or vehicle required to formulate an effective pharmaceutical composition.

[0062] With respect to a drug or pharmacologically active agent, the term "effective amount" or "therapeutically effective amount" refers to a non-toxic amount of the drug or agent sufficient to achieve the intended effect. For the oral dosage forms of the present invention, an "effective amount" of an active substance in the composition means the amount required to achieve the intended effect when used in combination with another active substance in the composition. The determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0063] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating a target disorder, disease, or condition.

[0064] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0065] Indicates a connection key.

[0066] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present invention. DETAILED DESCRIPTION

[0067] The present application is further described in detail below with reference to examples, but the implementation methods of the present application are not limited thereto.

[0068] Intermediate 1

[0069]

[0070] Synthesis of N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide. The synthetic route is as follows:

[0071]

[0072] Step A: tert-Butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate

[0073] At room temperature, (2R,3S)-2-((tert-Butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butanoic acid (3 g, 10.51 mmol) was dissolved in N,N-dimethylformamide (40 ml), and (2R)-1,2-dimethylpiperazine (1.2 g, 10.51 mmol) and N,N-diisopropylethylamine (11.58 ml, 70.08 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0074] Post-treatment: Dilute with saturated aqueous sodium bicarbonate (100 mL), extract with ethyl acetate (100 mL x 3), combine the organic phases, wash with saturated brine (50 mL x 2), dry over anhydrous sodium sulfate, filter, and concentrate to dryness to obtain 3.84 g of tert-butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate as a yellow oily liquid (yield 99%). LC-MS: [M+H] + =439.

[0075] Step B: (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one

[0076] To tert-butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate (3.84 g, 8.76 mmol) was dissolved in dichloromethane (20 ml) at room temperature, trifluoroacetic acid (16 ml, 219 mmol) was added, and the mixture was stirred at room temperature for 1 hour.

[0077] Post-treatment: Concentration, saturated aqueous sodium bicarbonate solution was added to the residue to adjust the pH to alkaline, and extraction was performed with ethyl acetate (100 mL x 3). The organic phases were combined and washed with saturated brine (50 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 2.96 g of (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one as a yellow oily liquid (yield 99%). LC-MS: [M+H]+ = 339.

[0078] Step C: Benzyl ((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate

[0079] At room temperature, (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one was dissolved in dichloromethane (50 ml), and benzyl 2,5-dioxopyrrolidin-1-yl carbonate (3.49 g, 14 mmol) and triethylamine (12.16 ml, 87.5 mmol) were added and reacted at room temperature overnight.

[0080] Post-treatment: Dilute with saturated aqueous sodium bicarbonate (100 mL), extract with dichloromethane (100 mL x 3), combine the organic phases, wash with saturated brine (50 mL x 2), dry over anhydrous sodium sulfate, filter, and concentrate to dryness to obtain 4.13 g of the product, benzyl ((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate (yield 99%) as a yellow oily liquid. LC-MS: [M+H] + =473.

[0081] Step D: Benzyl ((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate

[0082] Benzyl ((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate (4.13 methyl g, 0.13 mmol) was dissolved in ethanol (70 ml) and water (30 ml), iron powder (2.44 g, 43.7 mmol) and ammonium chloride (2.34 g, 43.7 mmol) were added, and the mixture was heated to 80 °C for 1 hour.

[0083] Post-treatment: filtration through celite, concentration, addition of saturated aqueous sodium bicarbonate solution (100 mL) to the residue, extraction with ethyl acetate (100 mL x 3), combination of the organic phases, washing with saturated brine (50 mL x 2), drying over anhydrous sodium sulfate, filtration, and concentration to dryness afforded 3.87 g of the product, benzyl ((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (yield 99%), as a yellow oily liquid. LC-MS: [M+H] + =443.

[0084] Step E: tert-Butyl ((S)-1-((4-((2S,3R)-3-((benzyloxy)carbonyl)amino)-4-(((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)carbamate

[0085] Benzyl ((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (3.87 g, 8.75 mmol) was dissolved in pyridine (60 ml) at room temperature, and (2S)-2-{[(tert-butoxy)carbonyl]amino}-3,3-dicyclopropylpropionic acid (3.06 g, 11.38 mmol) was added, and the mixture was stirred at room temperature for 1 hour.

[0086] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate solution (100 mL) to the residue, extraction with ethyl acetate (100 mL × 3), combination of the organic phases, washing with saturated brine (50 mL × 2), drying over anhydrous sodium sulfate, filtering, and concentration to dryness afforded 6.07 g of the product, tert-butyl ((S)-1-((4-((2S,3R)-3-((benzyloxy)carbonyl)amino)-4-(((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)carbamate (yield 99%). LC-MS: [M+H] + =694.

[0087] Step F: Benzyl ((2R,3S)-3-(4-(((S)-2-amino-3,3-dicyclopropylpropionamido)-3-fluorophenyl)-1-(((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate

[0088] At room temperature, tert-butyl (((S)-1-((4-((2S,3R)-3-((benzyloxy)carbonyl)amino)-4-(((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)carbamate (6.07 g, 8.75 mmol) was dissolved in dichloromethane (30 ml), trifluoroacetic acid (15 ml, 23.08 mmol) was added, and the mixture was stirred at room temperature for 1 hour.

[0089] Post-treatment: Concentration, saturated aqueous sodium bicarbonate solution was added to the residue to adjust the pH to alkaline, and extraction was performed with ethyl acetate (100 ml × 3). The organic phases were combined and washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 5.19 g of yellow oily liquid product, benzyl ((2R,3S)-3-(4-(((S)-2-amino-3,3-dicyclopropylpropionamido)-3-fluorophenyl)-1-(((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (yield 99%). LC-MS: [M+H] + =594.

[0090] Step G: Benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate

[0091] Benzyl ((2R,3S)-3-(4-(((S)-2-amino-3,3-dicyclopropylpropionamido)-3-fluorophenyl)-1-(((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (5.19 g, 8.74 mmol) was dissolved in N,N-dimethylformamide (40 ml) at room temperature, and 4-cyclopropyl-1,2,5-oxadiazole-3-carboxylic acid (1.48 g, 9.61 mmol), N,N-diisopropylethylamine (11.56 ml, 69.92 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.98 g, 13.11 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0092] Post-treatment: saturated aqueous sodium bicarbonate solution (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 6.38 g of the yellow oily liquid product, benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (yield 99%). LC-MS: [M+H]+ = 730.

[0093] Step H: N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide

[0094] Benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (6.07 g, 8.75 mmol) was dissolved in ethyl acetate (30 ml) at room temperature, and palladium on carbon (15 ml, 23.08 mmol) was added, followed by stirring at 50°C overnight.

[0095] Post-treatment: Pd-carbon was removed by filtration through celite, and the mixture was concentrated. The residue was dissolved in saturated sodium bicarbonate (100 mL) and extracted with ethyl acetate (100 mL x 3). The organic phases were combined and washed with saturated brine (50 mL x 2). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 5.2 g of the yellow solid product, N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (yield 99%). LC-MS: [M+H] + =596.

[0096] Intermediate 2

[0097]

[0098] Synthesis of (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid. The synthetic route is as follows:

[0099]

[0100] Step A: Methyl (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoate

[0101] Methyl (2R,3S)-3-(4-(((S)-2-amino-3,3-dicyclopropylpropionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoate (3.6 g, 8.01 mmol) was dissolved in N,N-dimethylformamide (25 ml) at room temperature, and 4-cyclopropyl-1,2,5-oxadiazole-3-carboxylic acid (1.48 g, 9.61 mmol), N,N-diisopropylethylamine (10.59 ml, 64.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.57 g, 12.02 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0102] Post-treatment: saturated aqueous sodium bicarbonate solution (100 ml) was added, extracted with ethyl acetate (100 ml × 3), the organic phases were combined, washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 4.69 g of yellow oily liquid product (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid methyl ester (yield 99%). LC-MS: [M+H] + =586.

[0103] Step B: (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid

[0104] Methyl (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoate (4.68 g, 8.0 mmol) was dissolved in a mixed solvent of tetrahydrofuran (50 ml) and methanol (30 ml) at room temperature. Lithium hydroxide monohydrate (0.67 g, 15.98 mmol) dissolved in water (15 ml) was added, and the mixture was stirred at room temperature for 1 hour.

[0105] Post-treatment: concentrated, diluted with water (15 ml), adjusted to acidic pH with 1 M aqueous hydrochloric acid solution, extracted with ethyl acetate (50 ml × 3), combined the organic phases, washed with saturated brine (50 ml × 1), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 4.57 g of brown solid product (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid. LC-MS: [M+H] + =572.

[0106] Intermediate 3

[0107]

[0108] Synthesis of (S)-2-amino-3,3-dicyclopropyl-N-(4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)propionamide

[0109]

[0110] Step A: tert-Butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate

[0111] At room temperature, (2R,3S)-2-((tert-Butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butanoic acid (2.3 g, 6.72 mmol) was dissolved in DMF (20 ml), and (2R)-1,2-dimethylpiperazine (0.92 g, 8.06 mmol), DIPEA (6.08 g, 47 mmol), and HATU (3.07 g, 8.06 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 h.

[0112] Saturated aqueous NaHCO₃ solution (10 mL) and 20 mL of water were added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and washed with saturated brine (50 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 0.23 g of tert-butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate as a light yellow solid (yield 100%). LC-MS: [M+H] + =438.

[0113] Step B: (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one

[0114] Tert-butyl (2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)carbamate (2.95 g, 6.73 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (17 g, 149 mmol) was added, and the mixture was reacted at room temperature for 1 h.

[0115] The filtrate was concentrated to obtain 2.28 g of a light yellow solid product (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one (yield 100%). LC-MS: [M+H] + =338.

[0116] Step C: N-((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)-2-methoxyacetamide

[0117] (2R,3S)-2-amino-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)butan-1-one (2.28 g, 6.74 mmol) was dissolved in DMF (8 ml), and methoxyacetic anhydride (1.09 g, 6.74 mmol) and DIPEA (3.48 g, 26.96 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 h.

[0118] Post-treatment: Add saturated aqueous NaHCO₃ solution (10 mL) and 20 mL of water, extract with ethyl acetate (30 mL x 3), combine the organic phases, wash with saturated brine (50 mL x 2), dry over anhydrous sodium sulfate, filter, and concentrate to dryness to obtain 1.91 g of yellow solid product, N-((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)-2-methoxyacetamide (yield 70%). LC-MS: [M+H] + =410.

[0119] Step D: N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)-2-methoxyacetamide

[0120] N-((2R,3S)-1-((R)-3,4-dimethylpiperazin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobutan-2-yl)-2-methoxyacetamide (1.91 g, 4.65 mmol) was dissolved in methanol (15 mL), palladium carbon (0.5 g, 10% wt) was added, H2 was replaced five times, and then hydrogen was separated and reacted at room temperature for 2 h.

[0121] The mixture was filtered, washed with 50 ml of methanol, and the filtrate was concentrated to obtain 1.7 g of a dark red solid product, N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)-2-methoxyacetamide (yield 80%). LC-MS: [M+H] + =380.

[0122] Step E: tert-Butyl ((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)carbamate

[0123] N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)-2-methoxyacetamide (700 mg, 1.84 mmol) was dissolved in pyridine (8 ml), and (S)-2-((tert-butoxycarbonyl)amino)-3,3-dicyclopropylpropionic acid (0.495 g, 1.84 mmol) and EDCI (1.05 g, 5.52 mmol) were added. After the addition, the mixture was reacted at room temperature for 3 h.

[0124] Post-treatment: 30 ml of water and 50 ml of a saturated aqueous sodium bicarbonate solution were added, and the mixture was extracted with EA (40 ml * 4). The organic phases were combined and washed with saturated brine (40 ml * 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 0.900 g of a yellow solid product ((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)carbamic acid tert-butyl ester (yield 77%). LC-MS: [M+H] + =631.

[0125] Step F: (S)-2-amino-3,3-dicyclopropyl-N-(4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)propanamide

[0126] Tert-butyl ((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)carbamate (0.9 g, 1.42 mmol) was dissolved in DCM (10 mL), and trifluoroacetic acid (5.8 g, 50.8 mmol) was added. After the addition was complete, the reaction was allowed to react at room temperature for 1 h.

[0127] Post-treatment: The reaction mixture was concentrated under reduced pressure and directly used in the next step. Concentration under reduced pressure afforded 0.470 g of a pale yellow solid product, (S)-2-amino-3,3-dicyclopropyl-N-(4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)propanamide (yield 63%). LC-MS: [M+H] + =531.

[0128] Intermediate 4

[0129]

[0130] Synthesis of N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-ethyl-1,2,5-oxadiazole-3-carboxamide

[0131] The specific synthetic route is as follows:

[0132]

[0133] Step A: Benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamide)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate

[0134] Benzyl ((2R,3S)-3-(4-(((S)-2-amino-3,3-dicyclopropylpropionamido)-3-fluorophenyl)-1-(((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (5.19 g, 8.74 mmol) was dissolved in N,N-dimethylformamide (40 ml) at room temperature, and 4-ethyl-1,2,5-oxadiazole-3-carboxylic acid (1.36 g, 9.61 mmol), N,N-diisopropylethylamine (11.56 ml, 69.92 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.98 g, 13.11 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0135] Post-treatment: saturated aqueous sodium bicarbonate solution (100 mL) was added, and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 6.27 g of the yellow oily liquid product, benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamide)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (yield 99%). LC-MS: [M+H]+ = 718.

[0136] Step B: N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-ethyl-1,2,5-oxadiazole-3-carboxamide

[0137] Benzyl ((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-1-((R)-3,4-dimethylpiperazin-1-yl)-1-oxobutan-2-yl)carbamate (6.27 g, 8.74 mmol) was dissolved in ethyl acetate (30 ml) at room temperature, and palladium on carbon (15 ml, 23.08 mmol) was added, followed by stirring at 50°C overnight.

[0138] Post-treatment: palladium on carbon was removed by filtration through algae, and the mixture was concentrated. The residue was dissolved in saturated sodium bicarbonate (100 mL) and extracted with ethyl acetate (100 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 2). The organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 5.1 g of a yellow solid product, N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-ethyl-1,2,5-oxadiazole-3-carboxamide (yield 99%). LC-MS: [M+H] + =584.

[0139] Example 40

[0140]

[0141] Synthesis of (2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]-N-(2-fluoro-4-[(2S,3R)-3-(2-methoxyacetamide)-4-(3-methyl-3,6-diazabicyclo[3.1.1]heptane-6-yl)-4-oxobutan-2-yl]phenyl)propanamide

[0142]

[0143] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (150 mg, 0.26 mmol) was dissolved in N,N-dimethylformamide (5 ml) at room temperature, 3-methyl-3,6-diazabicyclo[3.1.1]heptane trifluoroacetate (76.46 mg, 0.34 mmol), N,N-diisopropylethylamine (0.34 ml, 2.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (150 mg, 0.39 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0144] Post-treatment: Saturated aqueous sodium bicarbonate (20 mL) was added, and the mixture was extracted with ethyl acetate (25 mL x 3). The organic phases were combined and washed with saturated brine (20 mL x 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Preparative purification was performed to obtain 46 mg of the product (2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]-N-(2-fluoro-4-[(2S,3R)-3-(2-methoxyacetamide)-4-(3-methyl-3,6-diazabicyclo[3.1.1]hept-6-yl)-4-oxobutan-2-yl]phenyl)propanamide (yield 26%). LC-MS: [M+H]+ = 666. 1 H NMR (400MHz, DMSO-d6) δ9.98(d,J=7.8Hz,1H),9.14(d,J=8.9Hz,1H),8.19–7.67(m,2H),7.20–7.02(m,2H),5.03(d,J= 8.0Hz,1H),4.59–4.16(m,2H),4.12–3.90(m,1H),3.87(d,J=1.4Hz,2H),3.31(s,3H),3.09–2.85(m,1H),2.83–2.53(m, 2H),2.34–2.24(m,1H),2.18(s,2H),2.16–1.98(m,1H),1.91(d,J=5.7Hz,1.5H),1.70(ddd,J=25.6,16.9,8.6Hz,1.5H) ,1.25–1.10(m,6H),0.99(tt,J=5.0,1.8Hz,2H),0.93–0.69(m,3H),0.48(s,1H),0.43–0.36(m,1H),0.35–0.15(m,6H).

[0145] Example 41

[0146]

[0147] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-oxo-4-(2-oxa-7-azaspiro[3.5]non-7-yl)butan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide and 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3S)-3-(2-methoxyacetamide)-4-oxo-4-(2-oxa-7-azaspiro[3.5]non-7-yl)butan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The specific synthetic route is as follows:

[0148]

[0149] At room temperature, (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (130 mg, 0.23 mmol) was dissolved in EDCI (10 ml), and 2-oxa-7-azaspiro[3.5]nonane ester (59 mg, 46 mmol) and EDCI (0.18 g, 0.92 mmol) were added and stirred at room temperature for 2 hours.

[0150] Post-treatment: Concentrate, add saturated sodium bicarbonate aqueous solution (30 ml), extract with ethyl acetate (30 ml × L), combine the organic phases, wash the organic phase with saturated brine (30 ml × 1), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness to obtain a white product.

[0151] Example 41A: 4-Cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-oxo-4-(2-oxa-7-azaspiro[3.5]non-7-yl)butan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (40 mg, yield 25.83%).

[0152] LC-MS: [M+H] +=681.1H NMR (400MHz, DMSO-d6) δ9.75(s,1H),8.93(d,J=9.1Hz,1H),7.72(d,J=9.0Hz,1H),7.50(t,J=8.3Hz,1H),6.93(d,J=1 2.0Hz,1H),6.84(d,J=8.2Hz,1H),4.84–4.76(m,1H),4.72(t,J=9.3Hz,1H),4.06–3.96(m,2H),3.92–3.80(m,2H),3. 63(s,2H),3.18(s,1H),3.08(s,3H),2.99–2.87(m,2H),2.73(d,J=9.6Hz,1H),2.10–1.73(m,1H),1.40(s,2H),1.03– 0.97(m,5H),0.92(dd,J=8.3,2.5Hz,2H),0.82–0.73(m,3H),0.71–0.58(m,2H),0.57–0.47(m,1H),0.31–0.07(m,8H).

[0153] Example 41B: 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-oxo-4-(2-oxa-7-azaspiro[3.5]non-7-yl)butan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (20 mg, yield 12.91%). LC-MS: [M+H] + =681.

[0154] Example 44

[0155]

[0156] Synthesis of (2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]-N-(2-fluoro-4-[(2S,3R)-3-(2-methoxyacetamide)-4-(3-methyl-3,8-diazabicyclo[3.2.1]octan-8-yl)-4-oxobutan-2-yl]phenyl)propanamide

[0157]

[0158] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (150 mg, 0.26 mmol) was dissolved in N,N-dimethylformamide (5 ml) at room temperature. 3-Methyl-3,8-diazabicyclo[3.2.1]octane dihydrochloride (62.13 mg, 0.31 mmol), N,N-diisopropylethylamine (0.34 ml, 2.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (150 mg, 0.39 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0159] Post-treatment: Saturated aqueous sodium bicarbonate (20 mL) was added, and the mixture was extracted with ethyl acetate (25 mL × 3). The organic phases were combined and washed with saturated brine (20 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Preparative purification was performed to obtain 33 mg of a white solid product, (2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]-N-(2-fluoro-4-[(2S,3R)-3-(2-methoxyacetamide)-4-(3-methyl-3,8-diazabicyclo[3.2.1]octan-8-yl)-4-oxobutan-2-yl]phenyl)propanamide (yield 18%). LC-MS: [M+H]+ = 680. 1HNMR(400MHz,DMSO-d6)δ9.96(s,1H),9.14(t,J=8.0Hz,1H),8.06(d,J=8.8H z,0.5H),7.89(d,J=8.9Hz,0.5H),7.80(dt,J=40.8,8.3Hz,1H),7.19–7.03(m ,2H),5.03(ddd,J=9.2,6.9,3.7Hz,1H),4.79(t,J=9.5Hz,0.5H),4.64(t,J= 9.6Hz, 0.5H), 4.42–4.25 (m, 1H), 4.13 (dd, J=14.4, 6.5Hz, 1H), 3.86 (d, J=15. 1Hz,2H),3.29(s,2H),3.27–3.21(m,0.5H),3.17(s,1H),3.08(dd,J=9.9,6. 8Hz,0.5H),2.37(s,1H),2.32–2.20(m,1H),2.05(s,2H),1.90–1.82(m,2H),1 .72–1.45(m,3H),1.39–1.04(m,8H),0.99(tq,J=4.8,2.1Hz,2H),0.94–0.68( m,3H),0.48(d,J=7.7Hz,1H),0.38(dd,J=8.4,4.6Hz,1H),0.34–0.15(m,6H).

[0160] Example 45

[0161] Synthesis of N-((S)-1-((4-((2S,3R)-3-(2-chloroacetamide)-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide

[0162]

[0163] N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (120 mg, 0.20 mmol) was dissolved in DCM amine (20 ml) at room temperature, and TEA (120 mg, 1.2 mmol) was added, followed by the addition of 2-chloroacetyl chloride (34 mg, 0.30 mmol) at 0°C, and the mixture was stirred at room temperature for 1 hour.

[0164] Post-treatment: Concentrate, add saturated aqueous sodium bicarbonate solution (20 mL), extract with ethyl acetate (25 mL × 3), combine the organic phases, wash with saturated brine (20 mL × 1), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness. Preparative purification yields 70 mg of white solid product N-((S)-1-((4-((2S,3R)-3-(2-chloroacetamide)-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (yield 50%). LC-MS: [M+H] + = 672.1H NMR (400MHz, DMSO-d6) δ9.99(d,J=8.2Hz,1H),9.23–9.08(m,1H),8.73(t,J=7.8Hz,1H),7.80(dt,J=30. 0,8.3Hz,1H),7.26–7.02(m,2H),5.10–5.02(m,1H),4.99–4.87(m,1H),4.20–4.07(m,2H),4.05–3.64(m, 2H),3.17–3.03(m,1H),2.29(td,J=8.4,5.1Hz,1H),2.10(s,1H),1.93(s,3H),1.38–1.21(m,5H),1.15( dd,J=8.0,3.3Hz,2H),1.02(s,2H),0.96–0.73(m,7H),0.50(s,1H),0.41(s,1H),0.29(d,J=34.5Hz,6H).

[0165] Example 48

[0166]

[0167] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.2.0]octan-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0168] Step A: (1S,6R)-5-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylic acid tert-butyl ester

[0169] At room temperature, (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (300 mg, 0.37 mmol, Purity 70%), (1S,6R)-tert-butyl 2,5-diazabicyclo[4.2.0]octane-2-carboxylate (126.29 mg, 0.64 mmol) and DIPEA (379.97 mg, 2.94 mmol) were dissolved in DMF (3 mL), and HATU (279.47 mg, 0.73 mmol) was added, and the reaction was carried out at room temperature for 2 hours.

[0170] After the reaction was completed, water (50 mL) was added and the mixture was extracted with ethyl acetate (3*30 mL). The organic phases were combined, washed with saturated brine (3*20 mL), dried over anhydrous Na2SO4, filtered, and rotary evaporated to a residue. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 20 / 1, V / V) to give (1S,6R)-5-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.2.0]octan-2-carboxylic acid tert-butyl ester (300 mg, Yield 95.96%, Purity 90%) as a brown solid. LC-MS: [M+H-100]. + =666.

[0171] Step B: Synthesis of N-((S)-1-((4-((2S,3R)-4-((1R,6S)-2,5-diazabicyclo[4.2.0]oct-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide

[0172] At room temperature, tert-butyl (1S,6R)-5-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.2.0]octane-2-carboxylate (300 mg, 0.39 mmol) was dissolved in DCM (2 mL), and TFA (2347.46 mg, 20.59 mmol) was added, and the mixture was reacted at room temperature for 2 hours.

[0173] After the reaction is completed, the residue is concentrated by rotary evaporation. Dichloromethane was added to dissolve the product, and the mixture was placed in an ice bath. The pH was adjusted to approximately 8-9 with saturated NaHCO₃(aq). The mixture was extracted with dichloromethane (3*30 mL). The combined organic phases were washed with saturated brine (3*20 mL), dried over anhydrous Na₂SO₄, filtered, and rotary evaporated to a residue to give a yellow solid product, N-((S)-1-((4-((2S,3R)-4-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (240 mg, Yield 92.03%). LC-MS: [M+H] + =666.

[0174] Step C: Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.2.0]oct-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0175] At room temperature, N-((S)-1-((4-((2S,3R)-4-((1R,6S)-2,5-diazabicyclo[4.2.0]octan-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (240 mg, 0.36 mmol) and formaldehyde (56.90 mg, 0.72 mmol, Purity 38%) were dissolved in methanol (SO, 3 mL) and stirred at room temperature for half an hour. NaCNBH3 (45.24 mg, 0.72 mmol) was added and reacted at room temperature for 1 hour.

[0176] After the reaction was completed, water (30 ml) was added, and the mixture was extracted with ethyl acetate (3*30 mL). The organic phases were combined, washed with saturated brine (3*20 mL), and then dried over anhydrous Na2SO4, filtered and rotary evaporated to a residue. The resulting residue was purified by C18 reverse phase column chromatography (ACN / 0.05% NH3.H2O, 55% ACN) to obtain a white solid product 4-cyclopropyl-N-((S)-1,1-dicyclopropyl- 3-((2-Fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.2.0]oct-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (103 mg, Yield 42.03%), LC-MS: [M+H]+=680. 1 HNMR(400MHz,DMSO-d6)δ9.99(d,J=12.3Hz,1H),9.17(t,J=9.2Hz,1H),7.98(dd,J=86.8,8.9Hz,1H),7.77(dt,J=16.4,8.3Hz,1H),7.15–6.99(m,2H) ,5.05(s,1H),4.84(dt,J=41.2,9.3Hz,1H),4.41(dq,J=116.7,8.0Hz,1H), 4.08–3.95(m,1H),3.88(d,J=4.3Hz,2H),3.31(d,J=4.3Hz,3H),3.26–3.21 (m,1H),2.61(d,J=11.2Hz,2H),2.48–2.37(m,1H),2.30(d,J=5.2Hz,1H), 2.01(s,2H),1.81(dd,J=9.2,4.7Hz,2H),1.62(s,1H),1.23(dd,J=7.0,4.3 Hz,4H),1.15(dd,J=8.0,4.2Hz,2H),1.04–1.00(m,2H),0.93(s,1H),0.85( s,1H),0.75(q,J=8.5Hz,1H),0.50(s,1H),0.41(s,1H),0.37–0.17(m,8H).

[0177] Example 49

[0178]

[0179] Synthesis of [(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3R)-3,4-dimethylpiperazin-1-yl]-1-oxobutan-2-yl]-3-methoxypropionamide

[0180]

[0181] To (2S)-N-(4-[(2S,3R)-3-amino-4-[(3R)-3,4-dimethylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]propanamide (150 mg, 0.25 mmol) was dissolved in pyridine (3 ml) at room temperature, 3-methoxypropionic acid (31.23 mg, 0.3 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (143.77 mg, 0.75 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0182] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate solution (20 mL) to the residue, extraction with ethyl acetate (25 mL × 3), and the combined organic phases were washed with saturated brine (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to dryness. Preparative purification afforded 50 mg of the product, N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3R)-3,4-dimethylpiperazin-1-yl]-1-oxobutan-2-yl]-3-methoxypropionamide, as a white solid (yield 33%). LC-MS: [M+H]+ = 682. 1HNMR (400MHz, DMSO-d6) δ9.97(d,J=9.1Hz,1H),9.17(dd,J=9.0,5.2Hz,1H),8.36(t,J=9.1Hz,1H),7.76(dt,J= 31.9,8.3Hz,1H),7.18–6.97(m,2H),5.09–4.84(m,2H),4.04–3.63(m,2H),3.50(ddt,J=9.6,5.4,3.1Hz,2H),3. 20(s,3H),3.09–3.01(m,1H),2.43(dd,J=14.2,6.9Hz,1H),2.39–2.11(m,2H),1.99(d,J=68.5Hz,4H),1.34–1.0 8(m,6H),0.99(dq,J=7.5,3.4Hz,2H),0.94–0.67(m,7H),0.48(s,1H),0.39(q,J=4.7Hz,1H),0.35–0.15(m,6H).

[0183] Example 56

[0184]

[0185] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(1-methoxycyclopropane-1-carboxamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The synthetic route is as follows:

[0186]

[0187] N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (120 mg, 0.20 mmol) was dissolved in pyridine (20 ml) at room temperature, 1-methoxypropane-1-carboxylic acid (35 mg, 0.3 mmol) was added, and then EDCI (120 mg, 0.60 mmol) was added, and the mixture was stirred at room temperature for 2 hours.

[0188] Post-treatment: concentration, addition of saturated aqueous sodium bicarbonate solution (20 ml), extraction with ethyl acetate (25 ml × 3), combination of the organic phases, washing with saturated brine (20 ml × 1), drying the organic phase over anhydrous sodium sulfate, filtering, and concentrating the filtrate to dryness. Preparation and purification were performed to obtain 70 mg of a white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(1-methoxycyclopropane-1-carboxamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 42.93%). LC-MS: [M+H]+=694.1H NMR(400MHz,DMSO-d6)δ10.05(s,1H),9.24(s,1H),8.03(s,2H),7.17(s,2H),5.07(s ,2H),2.08(s,3H),1.94(s,3H),1.26(s,7H),1.03(s,7H),0.84(s,6H),0.24(s,7H).

[0189] Examples 57A and 57B

[0190]

[0191] Step A: tert-Butyl 5-((2R,3S)-3-(4-((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.1.0]heptane-2-carboxylate

[0192] At room temperature, 2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (400 mg, 0.49 mmol, Purity 70%), tert-butyl 2,5-diazabicyclo[4.1.0]heptane-2-carboxylate (126.29 mg, 0.64 mmol) and DIPEA (379.97 mg, 2.94 mmol) were dissolved in DMF (4 mL), and HATU (279.47 mg, 0.73 mmol) was added, and the reaction was carried out at room temperature for 2 hours.

[0193] After the reaction was completed, water (50 mL) was added and the mixture was extracted with ethyl acetate (3*30 mL). The combined organic phases were washed with saturated brine (3*20 mL), dried over anhydrous Na2SO4, filtered, and rotary evaporated to a residue. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 20 / 1, v / v) to give tert-butyl 5-((2R,3S)-3-(4-((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.1.0]heptane-2-carboxylate (400 mg, Yield 97.75%, Purity 90%) as a brown solid. LC-MS: [M+H-56]. + =694.

[0194] Step B: Synthesis of N-((2S)-1-((4-((2S,3R)-4-(2,5-diazabicyclo[4.1.0]heptane-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide

[0195] At room temperature, tert-butyl 5-((2R,3S)-3-(4-((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2,5-diazabicyclo[4.1.0]heptane-2-carboxylate (80 mg, 0.51 mmol) was dissolved in DCM (2 mL), and TFA (3070 mg, 26.93 mmol) was added, and the mixture was reacted at room temperature for 2 hours.

[0196] After the reaction was complete, the product was concentrated to a residue by rotary evaporation. Dichloromethane was added to dissolve the product, and the solution was placed in an ice bath. The pH was adjusted to approximately 8-9 with saturated NaHCO3 (aq). The mixture was extracted with dichloromethane (3*30 mL), and the organic phases were combined, washed with saturated brine (3*20 mL), and then dried over anhydrous Na2SO4. The product was filtered and rotary evaporated to a residue to obtain a yellow solid product, N-((2S)-1-((4-((2S,3R)-4-(2,5-diazabicyclo[4.1.0]heptane-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (300 mg, Yield 91.07%). LC-MS: [M+H] + =61.

[0197] Step C: Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-yl Formamide and 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1S,6R)-5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0198] At room temperature, N-((2S)-1-((4-((2S,3R)-4-(2,5-diazabicyclo[4.1.0]heptane-2-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (300 mg, 0.46 mmol) and formaldehyde (72.70 mg, 0.92 mmol) were dissolved in methanol (3 mL) and stirred at room temperature for half an hour. NaCNBH3 (57.81 mg, 0.92 mmol) was added and the reaction was allowed to react at room temperature for 1 hour.

[0199] After the reaction was completed, water (30 mL) was added, and the mixture was extracted with ethyl acetate (3*30 mL). The organic phases were combined, washed with saturated brine (3*20 mL), and then dried over anhydrous Na2SO4, filtered and rotary evaporated to a residue. The resulting residue was purified by C18 reverse phase column chromatography (ACN / 0.05% NH3.H2O, 55% ACN) to obtain a mixed product. The mixed product was subjected to chiral separation (chromatographic column: CHIRALCEL OD-H 4.6 mm x 250 mm x 5um; mobile phase-gradient condition Hex:EtOH=95:5), and the forward white solid product 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (20 mg, Yield 15.68%, Purity 4% 98%) and the latter peak white solid product 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((1S,6R)-5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (54 mg, Yield 41.90%, Purity 97%), LC-MS: [M+H] + =666.

[0200] NMR data of 57A or 57B: 1 H NMR (400MHz, DMSO-d6) δ10.01(d,J=11.4Hz,1H),9.17(t,J=8.9Hz,1H),8.05–7.61(m,2H),7.24–7.04(m,2 H),5.36–4.85(m,2H),3.97–3.80(m,3H),3.31(s,2H),3.18(dt,J=39.7,7.8Hz,1H),2.58(s,1H),2.40–2. 26(m,5H),2.11(s,1H),1.25(d,J=6.8Hz,4H),1.18–1.13(m,2H),1.01(tt,J=4.8,1.8Hz,2H),0.92(d,J=4 .7Hz,3H),0.75(q,J=8.9Hz,1H),0.70–0.56(m,1H),0.50(s,1H),0.41(d,J=3.9Hz,1H),0.38–0.18(m,8H).

[0201] NMR data of 57A or 57B: 1 H NMR (400MHz, DMSO-d6) δ10.02(s,1H),9.18(d,J=8.9Hz,1H),8.11(d,J=9.0Hz,1H),7.82(t,J=8.2Hz,1H),7.08(d,J=10 .2Hz,2H),5.15–4.98(m,2H),3.96–3.65(m,3H),3.32(d,J=4.4Hz,4H),3.17(t,J=7.1Hz,1H),2.44(q,J=7.2,6.7Hz,2H) ,2.33–2.25(m,1H),2.15(s,3H),1.61–1.50(m,1H),1.28(d,J=7.0Hz,4H),1.15(dd,J=8.4,3.1Hz,2H),1.01(td,J=5.2, 4.8,2.5Hz,2H),0.97–0.80(m,3H),0.75(q,J=8.6Hz,1H),0.50(s,1H),0.38(tt,J=14.1,6.6Hz,3H),0.30–0.17(m,5H).

[0202] Example 60

[0203]

[0204] Synthesis of N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-4-(2,2-difluoroethoxy)-1,2,5-oxadiazole-3-carboxamide

[0205]

[0206] At room temperature, (S)-2-amino-3,3-dicyclopropyl-N-(4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)propanamide (0.138 acyl g, 0.138 amide mmol) was dissolved in DMF (38 ml), and 4-(2,2-difluoroethoxy)-1,2,5-oxadiazole-3-carboxylic acid (0.050 g, 0.26 mmol), DIPEA (0.168 g, 1.3 mmol), and HATU (0.118 g, 0.31 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 h.

[0207] Saturated aqueous NaHCO₃ solution (10 mL) and 20 mL of water were added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. Liquid phase preparative purification was performed to obtain 42 mg of the product N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-4-(2,2-difluoroethoxy)-1,2,5-oxadiazole-3-carboxamide (yield 22%). LC-MS: [M+H] + =708. 1 HNMR(400MHz,DMSO-d6)δ10.05(d,J=9.4Hz,1H),9.12(t,J=7.5Hz,1H),7.97(dd,J=25.7,8.5Hz,1H),7.85–7.70(m,1H) ,7.18(t,J=10.9Hz,1H),7.09(dd,J=18.2,8.3Hz,1H),6.45(ddd,J=56.6,52.4,3.4Hz,1H),5.09–4.93(m,2H),4.80–4. 68(m,2H),3.88(s,3H),3.72(dd,J=45.3,13.2Hz,3H),2.10(d,J=2.7Hz,1H),1.94(s,2H),1.86(d,J=2.8Hz,1H),1.24( dd,J=14.8,6.8Hz,4H),0.86(d,J=6.1Hz,4H),0.81(d,J=6.3Hz,2H),0.73(s,1H),0.45–0.34(m,2H),0.31–0.18(m,5H).

[0208] Example 66

[0209]

[0210] Synthesis of N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)methylamino]propionamido]-3-fluorophenyl)-1-[(3R)-3,4-dimethylpiperazin-1-yl]-1-oxobutan-2-yl]propionamide

[0211]

[0212] To (2S)-N-(4-[(2S,3R)-3-amino-4-[(3R)-3,4-dimethylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]propionamide (300 mg, 0.5 mmol) was dissolved in DMF (3 ml) at room temperature, propionyl propionate (78.08 mg, 0.6 mmol) and DIPEA (0.41 ml, 2.5 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0213] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate (20 mL) to the residue, extraction with ethyl acetate (25 mL x 3), and the combined organic phases were washed with saturated brine (20 mL x 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to dryness. Purification was performed using a reverse-phase preparative column to obtain 68 mg of N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)methylamino]propionamido]-3-fluorophenyl)-1-[(3R)-3,4-dimethylpiperazin-1-yl]-1-oxobutan-2-yl]propionamide as a white solid (yield 21%). LC-MS: [M+H]+ = 652.

[0214] 1 H NMR(400MHz,DMSO-d6)δ9.96(d,J=9.0Hz,1H),9.14(dd,J=9.0,5.5Hz,1H),8.25(t,J=8.6Hz,1 H),7.76(dt,J=32.2,8.3Hz,1H),7.17–6.99(m,2H),5.08–4.98(m,1H),4.95–4.82(m,1H),4.0 3–3.62(m,2H),3.14–3.02(m,1H),2.34–2.04(m,5H),1.90(s,2.5H),1.35–1.09(m,6.5H),1.0 2–0.94(m,6H),0.93–0.69(m,7H),0.48(s,1H),0.38(dt,J=8.8,4.4Hz,1H),0.35–0.15(m,6H).

[0215] Example 72

[0216]

[0217] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S,-3-(methoxymethyl)-4-methylpiperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0218] Step 1: tert-Butyl [(2R, 3S)-3-(3-fluoro-4-nitrophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate

[0219] At room temperature, (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(3-fluoro-4-nitrophenyl)butanoic acid (3 g, 8.76 mmol) was dissolved in N,N-dimethylformamide (30 ml), and (2S)-2-(fluoromethyl)-1-methylpiperazine dihydrochloride (2.16 g, 10.51 mmol), N,N-diisopropylethylamine (11.58 ml, 70.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (5 g, 13.14 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0220] Post-treatment: Add saturated aqueous sodium bicarbonate (50 mL), extract with ethyl acetate (50 mL × L), combine the organic phases, wash with saturated brine (50 mL × L), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness to obtain the product, tert-butyl N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (4 g, yield 99%), as a yellow oily liquid. LC-MS: [M+H] + =457.

[0221] Step 2: tert-Butyl N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[(3S)-3-fluoromethyl-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate

[0222] At room temperature, tert-butyl (N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (4 g, 8.76 mmol) was dissolved in ethyl acetate (50 ml), and palladium on carbon (1.86 g, 17.52 mmol) was added. The hydrogen atmosphere was replaced three times, and the mixture was heated in an oil bath to 50°C for overnight.

[0223] Post-treatment: filtration through celite, concentration of the filtrate afforded a yellow oily liquid product, tert-butyl N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[(3S)-3-fluoromethyl-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (3.7 g, yield 99%). LC-MS: [M+H] + =427.

[0224] Step 3: Benzyl N-[(1S)-1-[(4-[(2S,3R)-3-{[(tert-butoxy)carbonyl]amino}-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)carbamoyl]-2,2-dicyclopropylethyl]carbamate

[0225] At room temperature, tert-butyl N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[(3S)-3-fluoromethyl-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (3.7 g, 8.68 mmol) was dissolved in pyridine (25 ml), and (2S)-2-{[(benzyloxy)carbonyl]amino}-3,3-dicyclopropylpropionic acid (3.42 g, 11.28 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (4.99 g, 26.04 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0226] Post-treatment: Concentrate, dilute the residue with 30 mL of saturated sodium bicarbonate solution, extract with ethyl acetate (30 mL*3), combine the organic phases, wash with saturated brine (30 mL*1), dry over anhydrous sodium sulfate, filter, and concentrate to obtain benzyl N-[(1S)-1-[(4-[(2S,3R)-3-{[(tert-butoxy)carbonyl]amino}-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)carbamoyl]-2,2-dicyclopropylethyl]carbamate as a yellow oily liquid (6.1 g, yield 98%). LC-MS: [M+H] + =712.

[0227] Step 4: tert-Butyl N-[(2R,3S)-3-(4-[(2S)-2-amino-3,3-dicyclopropylpropionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate

[0228] Benzyl N-[(1S)-1-[(4-[(2S,3R)-3-{[(tert-butoxy)carbonyl]amino}-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)carbamoyl]-2,2-dicyclopropylethyl]carbamate (6.1 g, 8.57 mmol) was dissolved in ethyl acetate (60 ml) at room temperature, and palladium on carbon (1.82 g, 17.14 mmol) was added. The hydrogen atmosphere was replaced three times, and the mixture was heated in an oil bath to 50°C for overnight.

[0229] Post-treatment: filtration through celite and concentration of the filtrate afforded a yellow oily liquid product, tert-butyl N-[(2R,3S)-3-(4-[(2S)-2-amino-3,3-dicyclopropylpropionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (4.95 g, yield 99%). LC-MS: [M+H] +

[0230] =578.

[0231] Step 5: tert-Butyl N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate

[0232] At room temperature, tert-butyl N-[(2R,3S)-3-(4-[(2S)-2-amino-3,3-dicyclopropylpropionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (920 mg, g 1.59 mmol) was dissolved in pyridine (20 ml), and 4-cyclopropyl-1,2,5-oxadiazole-3-carboxylic acid (290 mg, g 1.91 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (910 mg, g 4.77 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0233] Post-treatment: Concentrate, add saturated aqueous sodium bicarbonate solution (50 mL) to the residue, extract with ethyl acetate (50 mL × 3), combine the organic phases, wash with saturated brine (50 mL × 1), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness to obtain a yellow oily substance, tert-butyl N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (1.14 g, yield 99%). LC-MS: [M+H] + =714.

[0234] Step 6: (2S)-N-(4-[(2S,3R)-3-amino-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]propanamide

[0235] At room temperature, tert-butyl N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]carbamate (1.14 g, 1.60 mmol) was dissolved in dichloromethane (5 ml), trifluoroacetic acid (5 ml, 67.31 mmol) was added, and the reaction was carried out at room temperature for 1 hour.

[0236] Post-treatment: Silica was concentrated, and the residue was added with saturated aqueous sodium bicarbonate solution (50 mL). The mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness to obtain the yellow oily liquid product (2S)-N-(4-[(2S,3R)-3-amino-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]propanamide (0.79 g, yield 81%). LC-MS: [M+H] + =614.

[0237] Step 7: N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]-3-fluorooxetane-3-carboxamide

[0238] At room temperature, (2S)-N-(4-[(2S,3R)-3-amino-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamide]propanamide (200 mg, g 0.33 mmol) was dissolved in pyridine (3 ml), 3-fluorobutane-3-carboxylic acid (43.59 mg, g 0.36 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (189.78 mg, g 0.99 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0239] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate solution (50 mL) to the residue, extraction with ethyl acetate (50 mL × 3), combination of the organic phases, washing with saturated brine (50 mL × 1), drying over anhydrous sodium sulfate, filtering, and concentrating the filtrate to dryness to give N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]-3-fluorooxetane-3-carboxamide (65 mg, yield 28%). LC-MS: [M+H] + =716.

[0240] 1H NMR (400MHz, DMSO-d6) δ9.99(d,J=5.5Hz,1H),9.16(d,J=8.6Hz,1H),8.62(d,J=8.2Hz,1H),7.75(dt,J=15.7,8.2Hz,1H),7.1 8(dd,J=11.8,7.6Hz,1H),7.05(t,J=7.8Hz,1H),4.99(dt,J=36.5,9.3Hz,2H),4.74(tdd,J=28.8,14.6,6.7Hz,4H),4.48–4.21 (m,1H),4.02–3.71(m,2H),3.24(d,J=9.6Hz,1H),2.84(t,J=11.6Hz,1H),2.28(dd,J=8.6,4.6Hz,1H),2.09(d,J=70.2Hz,4H) ,1.26–1.10(m,7H),1.00(d,J=5.9Hz,2H),0.93–0.68(m,4H),0.47(d,J=8.9Hz,1H),0.39(d,J=3.8Hz,1H),0.35–0.16(m,7H).

[0241] Example 75A

[0242]

[0243] Synthesis of cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((R)-3-(methoxymethyl)-4-methylpiperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The specific synthetic route is as follows:

[0244]

[0245] Step 1: tert-Butyl-4-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2-(methoxymethyl)piperazine-1-carboxylate

[0246] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (190 mg, 0.33 mmol) was dissolved in N,N-dimethylformamide (2 ml) at room temperature. To this was added tert-butyl (2S)-2-(methoxymethyl)piperazine-1-carboxylate (91.20 mg, 0.40 mmol), N,N-diisopropylethylamine (0.44 ml, 2.64 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (190 mg, 0.49 mmol), and the mixture was stirred at room temperature for 1 hour.

[0247] Post-treatment: saturated aqueous sodium bicarbonate solution (30 mL) was added, extracted with ethyl acetate (30 mL × 3), the organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness to give the product (S)-4-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2-(methoxymethyl)piperazine-1-carboxylic acid tert-butyl ester (260 mg, yield 99%) as a yellow oily liquid. LC-MS: [M+H] + =784.

[0248] Step 2: Cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S,-3-(methoxymethyl)piperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0249] At room temperature, tert-butyl (S)-4-((2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-cyclopropyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoyl)-2-(methoxymethyl)piperazine-1-carboxylate (260 mg, 0.33 mmol) was dissolved in dichloromethane (2 ml), and trifluoroacetic acid (1.5 ml, 20.19 mmol) was added, followed by stirring at room temperature for 1 hour.

[0250] Post-treatment: Concentrate, add saturated aqueous sodium bicarbonate solution (30 mL) to adjust the pH to alkaline, extract with ethyl acetate (30 mL × 3), combine the organic phases, wash with saturated brine (30 mL × 1), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness to give the yellow oily liquid product 5-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S-3-(methoxymethyl)piperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (226 mg, yield 99%). LC-MS: [M+H] + =684.

[0251] Step 3: 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S,-3-(methoxymethyl)-4-methylpiperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0252] 5-Cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S,-3-(methoxymethyl)piperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (226 mg, 0.33 mmol) was dissolved in methanol (3 ml) at room temperature. 38% aqueous formaldehyde solution (52.16 mg, 0.66 mmol) was added first, and the mixture was stirred at room temperature for half an hour. Subsequently, sodium cyanoborohydride (41.47 mg, 0.66 mmol) was added, and the mixture was stirred at room temperature for 1 hour.

[0253] Post-treatment: Aqueous solution (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined and washed with saturated brine (30 mL × 1). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. The mixture was separated and purified using a C18 column to obtain a white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-3-(2-methoxyacetamide)-4-((S-3-(methoxymethyl)-4-methylpiperazin-1-yl)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (65 mg, 28% yield). LC-MS: [M+H] + =698.

[0254] 1 HNMR (400MHz, DMSO-d6) δ9.99(d,J=5.8Hz,1H),9.15(dd,J=9.1,3.2Hz,1H),7.92(dd,J=33.1,8.8Hz,1H),7.77(dt,J=16.5,8.3Hz,1H),7.18 –7.01(m,2H),5.08–4.86(m,2H),3.99–3.62(m,4H),3.30(s,3H),3.26 (s,1.5H),3.19–3.16(m,0.5H),3.15(s,1.5H),3.11(d,J=9.2Hz,0.5H) ,2.92–2.73(m,1H),2.59(d,J=11.6Hz,0.5H),2.46(d,J=8.8Hz,0.5H),2.27(tq,J=8.7,4.7Hz,1H),2.13(s,1.5H),1.98(s,1.5H),1.50–1.4 0(m,0.5H),1.26–1.09(m,7.5H),0.99(p,J=3.2Hz,2H),0.93–0.68(m, 4H),0.53–0.44(m,1H),0.39(dt,J=8.9,4.4Hz,1H),0.36–0.18(m,7H).

[0255] Example 75B

[0256]

[0257] Example 75B uses the same synthetic route as Example 75A, except that Example 75B uses (2R)-2-(methoxymethyl)piperazine-1-carboxylic acid tert-butyl ester as the starting material. LC-MS: [M+H] + =698. 1H NMR (400MHz, DMSO-d6) δ9.99(d,J=8.5Hz,1H),9.16(d,J=8.8Hz,1H),7.95(d,J=8.8Hz,1H),7.84–7.68(m,1H),7.19–6 .95(m,2H),5.04(t,J=7.7Hz,1H),4.96–4.88(m,1H),4.11–3.68(m,4H),3.30(s,3H),3.24(s,1H),3.17(s,2H),3.15– 3.05(m,1H),2.46–2.37(m,1H),2.28(dt,J=12.9,9.3Hz,2H),2.13(s,1H),1.96(s,2H),1.43(s,0.5H),1.26–1.10(m, 7.5H),0.99(q,J=3.7,2.9Hz,2H),0.92–0.69(m,4H),0.46(d,J=8.5Hz,1H),0.39(d,J=4.0Hz,1H),0.34–0.18(m,7H).

[0258] Example 76A

[0259]

[0260] Synthesis of (2R)-N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]-2-hydroxyoxetane-2-carboxamide

[0261]

[0262] To (2S)-N-(4-[(2S,3R)-3-amino-4-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-4-oxobutan-2-yl]-2-fluorophenyl)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)carboxamide]propanamide (250 mg, 0.41 mmol) was dissolved in pyridine (3 ml) at room temperature, (2R)-2-hydroheptene-2-carboxylic acid (50.23 mg, 0.49 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (235.79 mg, 1.23 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0263] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate solution (30 mL) to the residue, extraction with ethyl acetate (30 mL × 3), and the combined organic phases were washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to dryness. Purification was performed on a C18 column to obtain 94 mg of (2R)-N-[(2R,3S)-3-(4-[(2S)-3,3-dicyclopropyl-2-[(4-cyclopropyl-1,2,5-oxadiazol-3-yl)formamido]propionamido]-3-fluorophenyl)-1-[(3S)-3-(fluoromethyl)-4-methylpiperazin-1-yl]-1-oxobutan-2-yl]-2-hydroxyoxetane-2-carboxamide (yield 33%). LC-MS: [M+H]+ = 698.

[0264] 1 H NMR(400MHz,DMSO-d6)δ9.99(d,J=4.7Hz,1H),9.15(dd,J=9.0,3.3Hz,1H),8.07(dd, J=11.6,8.9Hz,1H),7.77(dt,J=11.6,8.3Hz,1H),7.21(dt,J=12.2,2.5Hz,1H),7.10( dt,J=8.9,2.5Hz,1H),5.03(td,J=9.1,7.0Hz,2H),4.93(ddd,J=9.1,6.5,1.2Hz,1H) ,4.67–4.57(m,2H),4.48–4.11(m,2H),4.02–3.72(m,2H),3.24(dt,J=13.1,8.2Hz,1H ),2.89(dtd,J=23.6,13.3,10.9,6.5Hz,2H),2.61–2.52(m,2H),2.48–2.36(m,1H),2 .27(tq,J=8.8,4.8Hz,1H),2.18(s,1H),2.08(d,J=8.7Hz,0.5H),2.02(s,2H),1.50–1 .39(m,0.5H),1.36–1.07(m,6H),0.99(qt,J=4.6,1.9Hz,2H),0.94–0.78(m,2H),0.8 1–0.68(m,1H),0.48(t,J=9.0Hz,1H),0.38(dt,J=8.5,4.2Hz,1H),0.36–0.17(m,6H).

[0265] Example 76B

[0266]

[0267] Example 76B was prepared using the same synthetic route as Example 76A, except that (2S)-2-hydroheptene-2-carboxylic acid was used as the starting material. LC-MS: [M+H]+=698. 1 H NMR (400MHz, DMSO-d6) δ9.98 (d, J=6.1Hz, 1H), 9.15 (dd, J=8.9, 3.2Hz, 1H), 8. 12(dd,J=20.6,8.7Hz,1H),7.75(dt,J=16.1,8.3Hz,1H),7.17(td,J=11.3,10. 4,1.9Hz,1H),7.10–7.02(m,1H),5.08–4.91(m,3H),4.59(qd,J=6.0,2.9Hz,1 H),4.55–4.12(m,3H),4.04–3.75(m,2H),3.20–3.09(m,1H),2.91–2.78(m,2H) ,2.68–2.53(m,2H),2.47–2.38(m,1H),2.27(tt,J=8.9,4.6Hz,1H),2.19(s,1 H),2.09(q,J=11.9,11.5Hz,0.5H),2.02(s,2H),1.47–1.37(m,0.5H),1.26–1. 08(m,6H),0.99(qd,J=5.0,4.3,1.9Hz,2H),0.94–0.78(m,2H),0.73(q,J=9.2, 8.8Hz,1H),0.52–0.45(m,1H),0.38(dt,J=8.8,4.3Hz,1H),0.35–0.17(m,6H).

[0268] Examples 77A and 77B

[0269]

[0270] Synthesis of 4-cyclopropyl-N-((2S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-4-(3-(fluoromethyl)-4-methylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide

[0271]

[0272] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (250 mg, 0.44 mmol) was dissolved in pyridine (5 ml) at room temperature. (2R)-2-(fluoromethyl)-1-methylpiperazine (108.28 mg, 0.53 mmol) and N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (253.04 mg, 1.32 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0273] Post-treatment: Concentration, addition of saturated aqueous sodium bicarbonate solution (40 mL) to the residue, extraction with ethyl acetate (40 mL × 3), and the combined organic phases were washed with saturated brine (40 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated to dryness. Purification was performed using a C18 preparative column to obtain two white products: 4-cyclopropyl-N-((2S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-4-(3-(fluoromethyl)-4-methylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide, the leading peak 1 (41 mg, 14% yield) and the trailing peak 2 (35 mg, 12% yield). LC-MS: [M+H] + =686.

[0274] 77A or 77B NMR data: 11H NMR (400 MHz, DMSO-d6) δ 9.99 (d, J = 16.3 Hz, 1H), 9.15 (t, J = 8.0 Hz, 1H), 8.01 (dd, J = 17.8, 8.8 Hz, 1H), 7.76 (dt, J = 24.8, 8.3 Hz, 1H), 7.20–7.00 (m, 2H), 5.07–4.89 (m, 2H), 4.49–4.22 (m, 2H), 4.03 (d, J = 12.0 Hz, 1H), 3.85 (s, 3H), 3.30 (s, 3H), 3.19–3.11 (m, 1H), 3.00 (t, J = 12.4 Hz, 0.6H), 2.68–2.59 (m, 0.4H), 2.44 (dd, J = 13.0, 10.5 Hz, 2H), 2.32–2.21 (m, 1H), 2.18 (s, 1H), 2.02 (s, 2H), 1.61–1.46 (m, 0.5H), 1.28–1.08 (m, 6.5H), 0.99 (tt, J = 5.9, 3.0 Hz, 2H), 0.86 (ddd, J = 28.0, 9.1, 4.9 Hz, 2H), 0.73 (q, J = 9.1, 8.6 Hz, 1H), 0.47 (d, J = 8.3 Hz, 1H), 0.38 (dt, J = 8.7, 4.2 Hz, 1H), 0.33–0.18 (m, 6H).

[0275] 1H NMR data of 77A or 77B: 1 1H NMR (400 MHz, DMSO-d6) δ 9.95 (d, J = 5.0 Hz, 1H), 9.14 (dd, J = 8.9, 2.4 Hz, 1H), 7.76–7.56 (m, 2H), 7.31–7.18 (m, 1H), 7.09 (dd, J = 16.8, 8.3 Hz, 1H), 5.08–4.96 (m, 2H), 4.69–4.38 (m, 2H), 4.16 (ddd, J = 31.2, 20.6, 12.7 Hz, 2H), 3.63 (s, 2H), 3.21 (dd, J = 17.3, 9.0 Hz, 1H), 3.07 (s, 3H), 2.85–2.70 (m, 2H), 2.33–2.22 (m, 4H), 2.06 (dd, J = 22.8, 10.4 Hz, 2H), 1.26–1.09 (m, 6H), 0.99 (tt, J = 4.9, 2.1 Hz, 2H), 0.93–0.78 (m, 2H), 0.79–0.68 (m, 1H), 0.51–0.47 (m, 1H), 0.38 (dt, J = 8.2, 4.0 Hz, 1H), 0.35–0.17 (m, 6H).

[0276] Example 99A

[0277]

[0278] Synthesis of N-((S)-1,1-dicyclopropyl-3-[(4-((2S,3R)-4-((R)-3,4-dimethyl <unk>(2-(4-(2-oxo-1-phenyl)-3-(2-methoxy-1-oxo ...

[0279]

[0280] Intermediate 4 (154 mg, 0.26 mmol) was dissolved in DMF (8 ml), and methoxyacetic anhydride (0.059 g, 0.31 mmol) and DIPEA (0.040 g, 0.31 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 h.

[0281] Post-treatment: add saturated NaHCO3 aqueous solution (10 ml), water 20 ml, extract with ethyl acetate (30 ml × 3), combine the organic phases, wash the organic phase with saturated brine (50 ml × 2), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate to dryness to obtain 0.07 g of N-((S)-1,1-dicyclopropyl-3-[(4-((2S,3R)-4-((R)-3,4-dimethyl <unk>1-oxazine-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-4-ethyl-1,2,5-oxadiazole-3-carboxamide (yield 40%). LC-MS: [M+H] + =655.

[0282] 1 H NMR (400MHz, DMSO-d6) δ9.95(d,J=8.5Hz,1H),9.10(dd,J=9.0,6.3Hz,1H),7.92(dd,J=25.6,8.8Hz,1H),7.75(dt,J=30.8,8.3Hz,1H ),7.18–7.08(m,1H),7.04(dd,J=15.8,8.4Hz,1H),5.05–4.88(m,2H),3.97(d,J=12.6Hz,1H),3.83(s,2H),3.62(d,J=13.2Hz,1H),3 .09(td,J=10.4,6.8Hz,1H),2.94–2.83(m,2H),2.70(t,J=11.6Hz,0H),2.14(dd,J=12.8,9.5Hz,0H),2.05(s,1H),1.88(s,2H),1.31 (dd,J=11.9,3.6Hz,1H),1.27(s,0H),1.21–1.14(m,3H),0.88(dq,J=8.4,4.2,3.3Hz,1H),0.84(s,0H),0.74(dd,J=19.4,7.3Hz,2H).

[0283] Example 100A

[0284]

[0285] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-(((R)-3,4-dimethylpiperazin-1-yl)-3-(((R)-2-methoxypropionamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The specific synthetic route is as follows:

[0286]

[0287] N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (95 mg, 0.16 mmol) was dissolved in pyridine (20 ml) at room temperature, and (R)-2-methoxypropionic acid (33 mg, 0.33 mmol) and EDCI (97 mg, 0.96 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0288] Post-treatment: Concentrate to remove pyridine, add saturated aqueous sodium bicarbonate solution (30 ml) and 30 ml of water, extract with ethyl acetate (25 ml × 3), combine the organic phases, wash the organic phase with saturated brine (20 ml × 1), dry the organic phase over anhydrous sodium sulfate, filter, and concentrate the filtrate to dryness. Preparative purification gives 40 mg of a white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-(((R)-3,4-dimethylpiperazin-1-yl)-3-(((R)-2-methoxypropionamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 21%). LC-MS: [M+H] + = 682.1H NMR(400MHz,DMSO-d6)δ9.98(d,J=9.0Hz,1H),9.21–9.13(m,1H),8.06–7.69(m,2H),7.29–7.00(m,2H),5.1 0–4.92(m,2H),4.04–3.63(m,3H),3.46(dd,J=7.0,5.1Hz,2H),3.33(s,3H),3.24(d,J=2.7Hz,3H),2.28(s, 1H),2.01(dt,J=13.4,7.1Hz,1H),1.94(s,2H),1.27–1.25(m,4H),1.22(dd,J=6.7,2.5Hz,4H),1.17–1.12( m,2H),1.01(s,2H),0.88–0.82(m,4H),0.79–0.72(m,1H),0.50(s,1H),0.41(s,1H),0.28(d,J=24.0Hz,6H).

[0289] Example 100B

[0290]

[0291] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-(((R)-3,4-dimethylpiperazin-1-yl)-3-(((S)-2-methoxypropionamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The specific synthetic route is as follows:

[0292]

[0293] N-((S)-1-((4-((2S,3R)-3-amino-4-((R)-3,4-dimethylpiperazin-1-yl)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropan-2-yl)-4-cyclopropyl-1,2,5-oxadiazole-3-carboxamide (95 mg, 0.16 mmol) was dissolved in pyridine (20 ml) at room temperature, and (S)-2-methoxypropionic acid (33 mg, 0.33 mmol) and EDCI (97 mg, 0.96 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0294] Post-treatment: Concentration to remove pyridine, addition of saturated aqueous sodium bicarbonate solution (30 ml) and 30 ml of water, extraction with ethyl acetate (25 ml × 3), combination of the organic phases, washing with saturated brine (20 ml × 1), drying over anhydrous sodium sulfate, filtering, and concentrating the filtrate to dryness. Preparation and purification afforded 45 mg of a white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-(((R)-3,4-dimethylpiperazin-1-yl)-3-(((S)-2-methoxypropionamido)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 41.39%). LC-MS: [M+H] + = 682.1H NMR (400MHz, DMSO-d6) δ9.99(d,J=8.6Hz,1H),9.17(d,J=8.8Hz,1H),8.12(dd,J=24.4,8.8Hz,1H),7.79(dt,J=30.2,8.3Hz,1H),7.23–7.01(m, 2H),5.05(q,J=7.7Hz,1H),4.94(dd,J=16.7,7.8Hz,1H),4.40–3.65(m,3H),3.46(dd,J=7.0,5.1Hz,1H),3.34(s,3H),3.24(s,3H),3.16(dd,J=1 7.4,9.1Hz,1H),2.29(td,J=8.5,4.8Hz,1H),2.10(s,1H),2.05–1.91(m ,3H),1.26(d,J=3.0Hz,3H),1.21–1.19(m,3H),1.18–1.13(m,2H),1.07( t,J=7.0Hz,2H),1.01(s,1H),0.87(d,J=2.4Hz,2H),0.82(d,J=6.3Hz,1 H),0.79–0.73(m,1H),0.50(s,1H),0.41(s,1H),0.29(d,J=34.1Hz,6H).

[0295] Example 102

[0296]

[0297] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-4-((S,-3-(fluoromethyl)-4-methylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The specific synthetic route is as follows:

[0298]

[0299] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (150 mg, 0.26 mmol) was dissolved in N,N-dimethylformamide (5 ml) at room temperature. (S)-2-(fluoromethyl)-1-methylpiperazine (41.24 mg, 0.31 mmol), N,N-diisopropylethylamine (0.34 ml, 2.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (148.29 mg, 0.39 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0300] Post-treatment: saturated aqueous sodium bicarbonate solution (20 ml) was added, extracted with ethyl acetate (25 ml × 3), the organic phases were combined, washed with saturated brine (20 ml × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Preparation and purification were performed to obtain 35 mg of a white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((2S,3R)-4-((S,-3-(fluoromethyl)-4-methylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)phenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 21%). LC-MS: [M+H] + = 686.1H NMR (400MHz, DMSO-d6) δ9.97(d,J=4.8Hz,1H),9.13(dd,J=9.0,3.3Hz,1H),7.96(dd,J=22.2,8.7Hz,1H),7.75(dt,J=1 4.1,8.3Hz,1H),7.16(ddd,J=10.8,8.6,1.9Hz,1H),7.11–7.02(m,1H),5.07–4.91(m,2H),4.50–4.05(m,2H),4.00–3.6 7(m,4H),3.31(s,2H),3.13(q,J=8.9,8.5Hz,1H),2.82(dd,J=13.4,10.5Hz,1H),2.54(s,2H),2.27(tt,J=8.9,4.6Hz, 1H), 2.09 (d, J = 70.4Hz, 4H), 1.44–0.68 (m, 12H), 0.48 (t, J = 9.0Hz, 1H), 0.38 (dt, J = 8.6, 4.1Hz, 1H), 0.36–0.15 (m, 6H).

[0301] Example 103A

[0302]

[0303] Compound 64, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide, was synthesized by the following synthetic route:

[0304]

[0305] (S)-2-Amino-3,3-dicyclopropyl-N-(4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamido)-4-oxobutan-2-yl)-2-fluorophenyl)propanamide (0.1 g, 0.19 mmol) was dissolved in DMF (8 ml) at room temperature, and 4-cyclopropyl-1,2,5-oxadiazole-3-carboxylic acid (32 mg, 0.21 mmol), DIPEA (0.25 g, 1.9 mmol) and HATU (0.094 g, 0.25 mmol) were added. After the addition, the mixture was stirred at room temperature for 1 h.

[0306] Saturated aqueous NaHCO₃ solution (10 mL) and 20 mL of water were added, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined and washed with saturated brine (50 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to dryness to obtain 55 mg of a pale white solid product, 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((R)-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 39.8%). LC-MS: [M+H] + =668.1H NMR(400MHz, DMSO-d6)δ10.00(d,J=8.3Hz,1H),9.19(dd,J=8.9,5.1Hz,1H), 8.06–7.68(m,2H),7.25–6.99(m,2H),5.00(dq,J=33.4,9.1,8.3Hz,2H),4.0 4–3.59(m,4H),3.31(d,J=1.7Hz,3H),3.20–3.04(m,1H),2.79–2.60(m,1H), 2.32–1.76(m,6H),1.37–1.10(m,7H),1.04–0.70(m,8H),0.61–0.10(m,8H).

[0307] Example 103B

[0308]

[0309] Synthesis of 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((S,-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2-yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide. The synthetic route is as follows:

[0310]

[0311] To (2R,3S)-3-(4-(((S)-3,3-dicyclopropyl-2-(4-ethyl-1,2,5-oxadiazole-3-carboxamido)propionamido)-3-fluorophenyl)-2-(2-methoxyacetamido)butanoic acid (150 mg, 0.26 mmol) was dissolved in N,N-dimethylformamide (4 ml) at room temperature. (S)-1,2-dimethylpiperazine (35.63 mg, 0.31 mmol), N,N-diisopropylethylamine (0.34 ml, 2.08 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (148.29 mg, 0.39 mmol) were added, and the mixture was stirred at room temperature for 1 hour.

[0312] Post-treatment: saturated aqueous sodium bicarbonate solution (20 ml) was added, extracted with ethyl acetate (25 ml × 3), the organic phases were combined, washed with saturated brine (20 ml × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness. Preparation and purification were performed to obtain 38 mg of a white solid product 4-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((2S,3R)-4-((S,-3,4-dimethylpiperazin-1-yl)-3-(2-methoxyacetamide)-4-oxobutan-2 -yl)-2-fluorophenyl)amino)-3-oxopropan-2-yl)-1,2,5-oxadiazole-3-carboxamide (yield 24%). LC-MS: [M+H]+ = 668. 1H NMR (400 MHz, DMSO-d6) δ 9.97 (d, J = 11.4 Hz, 1H), 9.14 (t, J = 8.2 Hz, 1H), 7.96 (dd, J = 30.1, 8.9 Hz, 1H), 7.77 (dt, J = 32.0, 8.3 Hz, 1H), 7.23–6 .98(m,2H),5.08–4.88(m,2H),4.08–3.70(m,4H),3.30(d,J=3.0Hz,3H),3.15(q,J=8.6,8.1Hz,1H),2.98(t, J=12.3Hz,0.5H),2.68–2.56(m,0.5H),2.47–2.32(m,1H),2.32–2.14(m,2H),2.01(d,J=57.8Hz,3H),1.80(d d,J=13.1,10.0Hz,0.5H),1.30(d,J=6.4Hz,0.5H),1.25–1.07(m,6H),0.99(ddt,J=7.4,5.2,2.2Hz,2H),0.9 4–0.68(m,6H),0.47(d,J=8.0Hz,1H),0.38(dt,J=9.0,4.3Hz,1H),0.26(dtd,J=26.6,16.2,15.7,6.5Hz,6H).

[0313] Comparative Example 1

[0314] The preparation of comparative example 1 was carried out with reference to patent document CN202080079537.5.

[0315] The synthetic routes of the compounds in Table 1 refer specifically to the synthetic routes of the compounds in the above examples and intermediates, except that the substituents of the intermediates are slightly different and are determined according to the target compound.

[0316] Table 1 shows the compound structures

[0317]

[0318]

[0319]

[0320]

[0321] Example 107 HT-29 cell human IL-17A neutralization experiment

[0322] HT-29 cells were used to evaluate the activity of the compounds in neutralizing human IL-17A at the cellular level. The specific experimental steps are as follows:

[0323] 1. Take HT-29 cells in the logarithmic growth phase, remove the original culture medium, rinse the cells with PBS, add trypsin to digest for 3 minutes, centrifuge (1000 × 3 minutes), discard the supernatant, resuspend in 3 ml, take out 100 μl and dilute 10 times for counting, and then use 2 × 10 5 Cells were seeded at 100 μg / ml in 96-well plates (#3596; Costar), and adherent cells were cultured for 24 h.

[0324] 2. Discard the original culture medium and add normal culture medium, culture medium containing different concentrations of compounds (starting concentration 1 μM, 4-fold dilution) and 90 ng / ml human IL-17A, and culture medium containing 90 ng / ml human IL-17A and different concentrations of secukinumab (starting concentration 10 nM, 5-fold dilution) to the plate.

[0325] 3. Continue incubation for 48 hours, then collect the supernatant.

[0326] 4: CXCL1 / GROa in the culture medium was detected using a commercial ELSIA kit (#DGR00B, R&D Systems). (The wells containing only culture medium were used as the bottom plate value for CXCL1 / GROa).

[0327] The inhibition rate was calculated using the following formula:

[0328]

[0329] 5. The experimental results were fitted and analyzed using Prism, and the IC50 values ​​of the compounds were calculated. The results are shown in Table 2.

[0330] Table 2 shows the inhibitory activity of the compounds against IL-17A

[0331]

[0332]

[0333] As shown in Table 2, the compounds of the present invention have good inhibitory activity against IL-17A.

[0334] Example 108

[0335] 1 Experimental Materials

[0336] SD rats: male, 180-250 g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0337] Reagents: DMSO (dimethyl sulfoxide), PEG-400 (polyethylene glycol 400), normal saline, heparin, acetonitrile, formic acid, propranolol (internal standard) are all commercially available. Instrument: AB SCIEX QTRAP 5500+.

[0338] 2 Experimental methods

[0339] Weigh the compound and dissolve it in DMSO-PEG-400-normal saline (5:60:35, v / v / v). After intravenous (iv) or oral gavage (ig) administration, collect 200 μL of venous blood into EDTA-K2 anticoagulant tubes at 15 minutes, 30 minutes, 1 hour, 2 hours, 5 hours, 7 hours, and 24 hours (5 minutes additionally for the iv group). Centrifuge at 12,000 rpm for 2 minutes, and freeze the plasma at -80°C for testing. Accurately weigh a certain amount of the test compound and dissolve it in DMSO to 2 mg / mL to prepare a stock solution. Accurately pipette an appropriate amount of the stock solution and dilute it with acetonitrile to prepare a series of standard solutions. Accurately pipette 10 μL of each of the above standard series solutions, add 90 μL of blank plasma, vortex mix, and prepare plasma samples equivalent to plasma concentrations of 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL. Perform double sample analysis for each concentration to establish a standard curve. Take 30 μL of plasma (plasma diluted 5 times 5 minutes, 15 minutes, and 30 minutes after intravenous administration), add 150 μL of acetonitrile solution of internal standard propranolol (50 ng / mL), vortex mix, add 100 μL of purified water, vortex mix again, centrifuge at 4000 rpm for 5 minutes, and take the supernatant for LC-MS analysis. LC-MS detection conditions are as follows:

[0340] Chromatographic column: YMC Triart C18, 50*3.0mm, 2.1μm.

[0341] Mobile phase: Water (0.1% formic acid)-acetonitrile for gradient elution as shown in Table 3:

[0342] Table 3

[0343] Time (min) A(％) B(％) 0 90％ 10％ 0.60 90％ 10％ 1.00 10％ 90％ 2.50 10％ 90％ 2.51 90％ 10％ 3.30 90％ 10％

[0344] 3 Data processing

[0345] After LC-MS detection of blood drug concentration, WinNonlin 6.1 software was used to calculate the pharmacokinetic parameters using the non-compartmental model method. The results are shown in Table 4.

[0346] Table 4

[0347]

[0348] As shown in Table 4, the compound of the present invention has a better in vivo exposure compared with Comparative Example 1.

[0349] The above embodiments are preferred implementations of the present invention, but the implementations of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered equivalent replacement methods and are included in the scope of protection of the present invention.< / unk> < / unk>

Claims

1. A compound represented by formula (I) or its racemate, isomer, or pharmaceutically acceptable salt, characterized in that: Wherein, n is 0, 1, 2, 3 or 4; Ring A is selected from R 1 Substituted heterocycloalkyl, aryl, heteroaryl, the R 1 is selected from alkyl, halogen, cycloalkyl, haloalkyl, haloalkoxy, halocycloalkyl, -O-cycloalkyl; R 2 is selected from substituted or unsubstituted cycloalkyl, heterocycloalkyl, aryl, heteroaryl, -CH(R 2’ )(R 2” ), the substituent is selected from alkyl, halogen, haloalkyl, haloalkenyl, the R 2’ and R 2” are independently selected from alkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl; R 3 is selected from hydrogen, halogen, and alkyl; R 4 is selected from hydrogen, halogen, alkyl, cycloalkyl; X 1 、X 2 solely selected from hydrogen, Or, X 1 、X 2 Cyclize together to form a substituted or unsubstituted heterocycloalkyl or heteroaryl, wherein the substitution is selected from alkyl, halogen; R 5a 、R 5b independently selected from alkyl, alkenyl, alkynyl, nitrile, alkylnitrile, haloalkyl, haloalkenyl, or substituted or unsubstituted -(CH2) k -O-alkyl, -(CH2) p -O-cycloalkyl, -(CH2) q B1B2-O-alkyl, cycloalkyl, heterocycloalkyl, alkylheterocycloalkyl, halogenated heterocycloalkyl, aryl, heteroaryl, the substituent is selected from alkyl, halogen, alkoxy, B1 and B2 are independently selected from alkyl and halogen, or B1 and B2 are cyclized together to form a substituted or unsubstituted cycloalkyl or heterocycloalkyl, the substituent is selected from alkyl and halogen; R 6 and R 7 independently selected from alkyl, alkoxy, cycloalkyl, Or, R 6 and R 7 Cyclize together to form a substituted or unsubstituted heterocycloalkyl, wherein the substitution is selected from alkyl, alkyl alcohol, alkoxy, alkoxyamide, halogen, cycloalkyl, -(CH2) k -O-alkyl, haloalkyl; k, m, p, q, y are independently selected from 1, 2, 3, 4, 5 or 6; Among them, when R 5a Selected from -CH2-O-alkyl or alkyl, said R 6 and R 7 Cyclize together When the A ring is not When R 5a When selected from alkyl, the R 6 and R 7 Cyclize together to form a substituted heterocycloalkyl, wherein the substitution is selected from alkoxyamide; When R 5a is selected from alkyl groups, wherein R 6 and R 7 Cyclize together When the R 2 for When R 5a When selected from unsubstituted heteroaryl, the R 6 and R 7 Cyclize together When R 5a Selected from When the R 2 Not for When R 2 Selected from When n is not 0 or 1, or R 6 and R 7 Cyclize together to form a heterocycloalkyl group substituted by halogen.

2. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that The alkyl group is selected from C 1-6 The alkyl group, the C 1-6 The alkyl group is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, 1-ethylpropyl, 2-methylbutyl, tert-pentyl, 1,2-dimethylpropyl, isopentyl, neopentyl, n-hexyl, isohexyl, sec-hexyl, tert-hexyl, neohexyl, 2-methylpentyl, 1,2-dimethylbutyl, and 1-ethylbutyl; The alkenyl group is selected from C 2-6 Alkenyl, C 2-6 The alkenyl group is selected from the group consisting of vinyl, 1-propenyl, 2-propenyl, 1-methylvinyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1-methyl-2-propenyl, 2-methyl-2-propenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-methyl-1-butenyl, 2-methyl-1-butenyl, 3-methyl-1-butenyl, 1-methyl-2-butenyl, 2-methyl-2-butenyl, 3-methyl-2-butenyl, 1-methyl-3-butenyl, 2-methyl-3-butenyl. , 3-methyl-3-butenyl, 1,1-dimethyl-2-propenyl, 1,2-dimethyl-1-propenyl, 1,2-dimethyl-2-propenyl, 1-ethyl-1-propenyl, 1-ethyl-2-propenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-methyl-1-pentenyl, 2-methyl-1-pentenyl, 3-methyl-1-pentenyl, 4-methyl-1-pentenyl, 1-methyl-2-pentenyl, 2-methyl-2-pentenyl, 3-methyl-2-pentenyl, 4-methyl-2-pentenyl, 1-methyl-3-pentenyl 1-methyl-4-pentenyl, 2-methyl-4-pentenyl, 3-methyl-4-pentenyl, 4-methyl-4-pentenyl, 1,1-dimethyl-2-butenyl, 1,1-dimethyl-3-butenyl, 1,2-dimethyl-1-butenyl, 1,2-dimethyl-2-butenyl, 1,2-dimethyl-3-butenyl, 1,3-dimethyl-1-butenyl, 1,3-dimethyl-2-butenyl, 1,3-dimethyl-3-butenyl, 2,2-dimethyl-3-butenyl 1-ethyl-2-methyl-1-propenyl, 1-ethyl-2-methyl-2-propenyl, 1-ethyl-3-butenyl, 2-ethyl-1-butenyl, 2-ethyl-2-butenyl, 2-ethyl-3-butenyl, 1,1,2-trimethyl-2-propenyl, 1-ethyl-1-methyl-2-propenyl, 1-ethyl-2-methyl-1-propenyl, and 1-ethyl-2-methyl-2-propenyl; The alkynyl group is selected from C 2-6 The alkynyl group, the C 2-6 The alkynyl group is selected from ethynyl, propynyl, 2-butynyl, 2-pentynyl, 3-pentynyl, 2-hexynyl, 3-hexynyl; The alkyl nitrile is selected from acetonitrile, propionitrile, butyronitrile, isobutyronitrile, valeronitrile, isovaleronitrile, and hexanenitrile; The aryl group is selected from a five-membered or six-membered aryl group; the heteroaryl group means that at least one carbon atom on the aryl group is replaced by a heteroatom, and is further preferably 3. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that The cycloalkyl group is selected from C 3-12 Cycloalkane, C 3-12 The cycloalkyl is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, bicyclic alkyl, bridged cycloalkyl, and spirocyclic alkyl; the bicyclic alkyl means that two rings share two carbon atoms, the bridged cycloalkyl means that two rings share two or more carbon atoms, the spirocyclic alkyl means that two cycloalkyls share one carbon atom, the heterocycloalkyl means that at least one carbon atom on the cycloalkyl is substituted by a heteroatom, the alkylheterocycloalkyl means that at least one hydrogen atom on the heterocycloalkyl is substituted by an alkyl, and the halogenated heterocycloalkyl means that at least one hydrogen atom on the heterocycloalkyl is substituted by a halogen.

4. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that The halogen is selected from fluorine, chlorine, bromine, and iodine; a haloalkyl group refers to an alkyl group in which at least one hydrogen atom is replaced by a halogen; and the heteroatom is selected from nitrogen, oxygen, and sulfur, and the heteroatom is one or more.

5. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that The bicyclic alkyl group is selected from: The bridged cycloalkyl group is selected from: The spiroalkyl group is selected from:

6. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that: The A ring is selected from The R 1 The substituted A ring is selected from R 2 Selected from cycloheptyl, n is 0 or 1, m, y are 1, k is 1 or 2, p, q are 1; R 3 is selected from hydrogen, methyl or fluorine; R 4 is selected from hydrogen or methyl; X 1 、X 2 solely selected from hydrogen, or X 1 、X 2 Cyclize together R 5a Selected from ethyl, vinyl, ethynyl, propadiene, nitrile, acetonitrile, R 5b is selected from methyl, ethyl, cyclopropyl, and cyclobutyl; R 6 and R 7 independently selected from methyl, methoxy, methoxymethyl, or R 6 and R 7 Cyclize together And when R 5a Selected from When R 6 and R 7 Not cyclized together When R 5a When selected from ethyl, R 6 and R 7 Cyclize together When R 5a When selected from ethyl, the R 6 and R 7 Cyclize together When the R 2 for When R 5a Selected from When the R 6 and R 7 Cyclize together 7. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that A compound selected from the group consisting of: a compound represented by formula (Ia), a racemate thereof, an isomer thereof, or a pharmaceutically acceptable salt thereof: Where n is 0 or 1, R 2 Selected from cycloheptyl, A ring, R 3 、X 1 、X 2 、R 6 、R 7 As defined above.

8. The compound according to claim 1, or its racemate, or its isomer, or its pharmaceutically acceptable salt, characterized in that: Select from the structures shown in Table 1.

9. A pharmaceutical composition, characterized in that The invention comprises a therapeutically effective amount of the compound according to any one of claims 1 to 8, or a racemate, an isomer, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

10. Use of the compound according to any one of claims 1 to 8, or its racemate, or its isomer, or its pharmaceutically acceptable salt, or the pharmaceutical composition according to claim 9 in the preparation of a medicament for treating a disease, characterized in that: The disease is an IL-17A-related disease, specifically selected from psoriasis and arthritis.

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