IL-17A related immunomodulator and application thereof

Abstract

The invention belongs to the field of medicine, and relates to an IL-17A related immunomodulator and application thereof, and application of the compound in treating autoimmune diseases or inflammatory diseases, in particular to a compound shown in the formula I, or an isomer or pharmaceutically acceptable salt thereof and application thereof, and substituent groups in the formula I are as defined in the specification.

Description

[0001] This invention claims priority to an earlier application filed on August 15, 2024, with China National Intellectual Property Administration, patent application number 202411121108.6, entitled "An IL-17A-related immunomodulator and its use therein". The entire contents of the aforementioned earlier application are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of medicine, specifically relating to an IL-17A-related immunomodulator and its uses, as well as the use of such compounds in the treatment of autoimmune diseases or inflammatory diseases. Background Technology

[0003] IL-17 (interleukin-17) is a pro-inflammatory cytokine. IL-17 secretion can stimulate the production of other pro-inflammatory cytokines (IL-1, IL-6, G-CSF, GM-CSF, and TNF), chemokines (CXCL1, CXCL2, CXCL5, CCL2, CCL7, CCL20, and IL-8), matrix metalloproteinases (MMP1, MMP3, MMP9, and MMP13), and antimicrobial peptides (β-defensins, S-100 protein) (Frontiers in Immunology (2020) 11: 947). IL-17 is composed of IL-17A (CTLA-8), IL-17B, IL-17C, IL-17D, IL-17E (IL-25), and IL-17F. IL-17A, as the most widely studied early member, is primarily concerned with its role in host defense mechanisms against various microbial pathogens and tissue inflammation. Human IL-17A is a glycoprotein with a molecular weight of approximately 17,000 Daltons. IL-17F, the closest relative to IL-17A, shares about 50% amino acid sequence homology with IL-17A in its 163 amino acids and is often co-expressed with IL-17A. Both IL-17A and IL-17F are secreted by helper T cells (Th17) and expressed as homodimers or IL-17A / F heterodimers (Eur opean Respiratory Journal (2005): 159-172). The IL-17 receptor family consists of five members: IL-17RA, IL-17RB, IL-17RC, IL-17RD, and IL-17RE. All five receptors share a common cytoplasmic motif called the SEFI R domain (Frontiers in Immunology 11 (2020): 947). IL-17A delivers signals into cells via the IL-17 receptor complex (IL-17RA and IL-17RC). The primary function of IL-17A is to coordinate local tissue inflammation through the upregulation of pro-inflammatory and neutrophil-migrating cytokines and chemokines (including IL-6, G-CSF, TNF-α, IL-1, CXCL1, CCL2, and CXCL2), and the penetration of the extracellular matrix by matrix metalloproteinase-activated T cells. Studies have shown that IL-17A plays a significant role in severe asthma and chronic obstructive pulmonary disease (COPD), patients who are often unresponsive or poorly responsive to currently available medications (Al-Ramli et al. J Allergy Clin Immunol, 2009, 123:1185-1187).Upregulation of IL-17A levels is involved in many diseases, including rheumatoid arthritis (RA), bone erosion, intraperitoneal abscess, inflammatory bowel disease, allogeneic graft rejection, psoriasis, atherosclerosis, asthma, and multiple sclerosis. Targeting both IL-17A and IL-17RA is an effective strategy for treating IL-17A-mediated autoimmune inflammatory diseases.

[0004] IL-17A antibody drugs have shown promising results in IL-7A-mediated inflammatory diseases, including asthma, psoriasis, rheumatoid arthritis, ankylosing spondylitis, and multiple sclerosis. Currently, several IL-17A-targeting antibodies have been FDA-approved for the treatment of moderate to severe plaque psoriasis. These antibodies include: secukinumab (Cosentyx, Novartis), ixekizumab (Taltz, Eli Lilly), brodalumab (Kyntheum, LEO Pharma / AstraZeneca), and bimekizumab (UCB).

[0005] Despite the existence of various IL-17A antibodies, small-molecule specific inhibitors of IL-17 with oral bioavailability are rarely reported. Several small molecules with IL-17A inhibitory activity have been disclosed (WO2020163554A1, WO2021055376A1, WO2023283453A1). Given the limitations of antibody drugs (high commercial cost, non-oral administration, poor tissue penetration, and typically long half-life), the development of small-molecule IL-17A inhibitors holds great promise. Summary of the Invention

[0006] A first aspect of the present invention provides a compound of formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] Where: n is selected from 0 or 1;

[0009] When n = 0, ring A is selected from an optional substituted 5-membered heteroaromatic ring, wherein ring A is independently substituted by one or more substituents selected from the following: C 1-6 Alkyl and C 3-5 cycloalkyl; wherein the C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, halogen, and C. 3-6 cycloalkyl;

[0010] When n=1, ring A is selected from an optionally substituted benzene ring or a 6-membered heteroaromatic ring, wherein the 6-membered heteroaromatic ring is an optionally substituted 6-membered heteroaromatic ring containing 1 or 2 nitrogen atoms; wherein ring A is independently substituted by one or more substituents selected from the following: halogen, -OR 11 -CN, -NHS(O)2CH3 and C 1-6 Alkyl; wherein the C 1-6 The alkyl group may optionally be substituted with one or more of the following substituents: hydrogen and halogen;

[0011] R 11 Independently selected from: hydrogen and C 1-6 Alkyl, wherein the C 1-6 The alkyl group is independently substituted by one or more substituents selected from the following: hydrogen and halogen;

[0012] R' and R'' are independently selected from: F and methyl; or R' and R'' can be linked together to form a cyclopropyl group;

[0013] R 1 and R 2 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen;

[0014] or the R 1 and R 2 C formed by combining with the same carbon 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is substituted with one or more substituents selected from the following: hydrogen and halogen;

[0015] R 3 Selected from: methyl, halomethyl, ethyl, haloethyl, and methoxymethyl; R 4 Selected from hydrogen;

[0016] R 5 Selected from methyl, halomethyl, ethyl, methoxymethyl, haloethyl, and:

[0017] X is selected from: C(R) 10 ), nitrogen, oxygen, and sulfur; Y and Z are each independently selected from: nitrogen, oxygen, and sulfur; wherein R 10 Selected from: hydrogen, fluorine and C 1-6 alkyl;

[0018] m is selected from 0, 1, or 2;

[0019] R 6 R 7 Each element is independently selected from hydrogen, halogens, and carbon. 1-6 Alkyl and C 1-6 Halogenated alkyl; or the R 6 and R 7 They can connect and combine to form C 3-7 cycloalkyl;

[0020] R 8 and R 9 Each is independently selected from hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen, halogen, C 1-6 Alkyl and C 3-7 cycloalkyl;

[0021] Or R 8 and R 9 They can be linked together to form 3 to 7-membered mono-heterocyclic alkyl groups, 5 to 12-membered bridged-ring heterocyclic alkyl groups, or 5 to 12-membered spirocyclic heterocyclic alkyl groups. The 3- to 7-membered monoheterocyclic alkyl, 5- to 12-membered bridged-ring heterocyclic alkyl, or 5- to 12-membered spirocyclic heterocyclic alkyl are mentioned. Each may be optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen and halogen.

[0022] In some embodiments, when n = 0, ring A is selected from:

[0023]

[0024] In some embodiments, when n=1, ring A is selected from:

[0025]

[0026] In some embodiments, R 1 and R 2 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen;

[0027] in Selected from:

[0028] In some embodiments, R 1 and R 2 Combines with the same carbon to form C 5-7 Cycloalkyl group, the C 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is substituted with one or more substituents selected from: hydrogen and halogen; which are selected from:

[0029]

[0030] In some embodiments, R 1 and R 2 Combines with the same carbon to form C 5-7 Cycloalkyl group, the C 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-3 Alkyl groups; selected from:

[0031]

[0032] In some embodiments, R 3 Selected from methyl; R 4 Selected from hydrogen.

[0033] In some embodiments, R 5 Selected from ethyl, methoxymethyl and:

[0034] In some embodiments, Selected from:

[0035]

[0036] In some embodiments, m is selected from 1.

[0037] In some embodiments, R 6 and R 7 Each is independently selected from hydrogen, fluorine, methyl, and trifluoromethyl; or the R 6 and R 7 They can combine to form cyclopropane.

[0038] In some embodiments, R 8 and R 9 Each is independently selected from hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl;

[0039] Wherein C 1-6Alkyl and C 3-7 The cycloalkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen, halogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein Selected from:

[0040]

[0041] In some embodiments, R 8 and R 9 They can be linked together to form 3 to 7-membered mono-heterocyclic alkyl groups, 5 to 12-membered bridged-ring heterocyclic alkyl groups, or 5 to 12-membered spirocyclic heterocyclic alkyl groups.

[0042] The 3- to 7-membered monoheterocyclic alkyl, 5- to 12-membered bridged-ring heterocyclic alkyl, or 5- to 12-membered spirocyclic heterocyclic alkyl are mentioned. Each may be optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 alkyl;

[0043] Wherein C 1-6 The alkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen;

[0044] The above Selected from:

[0045]

[0046] In some embodiments, the Selected from:

[0047]

[0048] In some embodiments, the compound of formula I is represented by formula II:

[0049]

[0050] Among them, rings A and R 1 R 2 R 5 R 6 R 7 R 8 R 9 R', R'', m, n, X, Y and Z are as described in the first aspect of the invention.

[0051] In some embodiments, the compound is represented by formula III:

[0052]

[0053] Among them B1 B 2 B 3 B 4 and B 5 Each is independently selected from: CR 16 and nitrogen;

[0054] Wherein R 16 Selected from: hydrogen, halogen, -OR 11 -CN, -NHS(O)2CH3 and C 1-6 alkyl;

[0055] And B 1 B 2 B 3 B 4 and B 5 No more than two of them are selected from nitrogen, and the rest are CR. 16 ;

[0056] R 1 R 2 R 5 R 6 R 7 R 8 R 9 R 11 X, Y, and Z are as described in the first aspect of the present invention.

[0057] In some embodiments, the compound is represented by formula IV:

[0058]

[0059] Where C 1 C 2 C 3 and C 4 Each is independently selected from: CR 17 NR 18 N and oxygen;

[0060] Wherein R 17 and R 18 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-5 cycloalkyl;

[0061] Wherein C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, halogen, and C. 3-6 cycloalkyl;

[0062] And C 1 C 2 C 3 and C 4 At least one of them is selected from NR18 , N or oxygen, the rest are CR 17 ;

[0063] R 1 R 2 R 5 R 6 R 7 R 8 R 9 X, Y, and Z are as described in the first aspect of the present invention.

[0064] In some embodiments, the compound is represented by formula V:

[0065]

[0066] Where R 18 Selected from C 1-6 Alkyl and C 3-5 cycloalkyl, wherein the C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; R 1 R 2 R 5 R 6 R 7 R 8 R 9 X, Y, and Z are as described in the first aspect of the present invention.

[0067] In some embodiments, the compound is selected from any of the following compounds:

[0068]

[0069]

[0070] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound, a stereoisomer or a pharmaceutically acceptable salt thereof as described in the first aspect, and one or more pharmaceutically acceptable carriers, diluents or excipients.

[0071] The third aspect of the invention is the use of the compound or a pharmaceutically acceptable salt thereof, or a solvate, metabolite, cocrystal or prodrug thereof, as described in the first aspect, or the pharmaceutical composition described in the second aspect, in the preparation of a medicament for treating / preventing IL-17A-mediated diseases.

[0072] The diseases mentioned include plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, erythrodermic psoriasis, and palmoplantar psoriasis. Detailed Implementation

[0073] Detailed description of the invention

[0074] Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0075] "Alkyl" refers to a saturated aliphatic hydrocarbon group. It includes straight-chain or branched groups with 1 to 20 carbon atoms. Preferably, it is a medium-sized alkyl group containing 1 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, etc. The alkyl group can be substituted or unsubstituted.

[0076] “C x-y "Intended to include groups containing x to y carbons in the chain. For example, the term "C..." 1-6 "Alkyl" refers to a saturated hydrocarbon group, including straight-chain alkyl and branched-chain alkyl containing 1 to 6 carbons.

[0077] “C x-y "Alkenyl" and "C" x-y "Alkyne" refers to an unsaturated aliphatic group whose length and possible substitutions are similar to those of the alkyl groups mentioned above, but each contains at least one double or triple bond.

[0078] “Cycloalkyl” refers to a saturated carbide ring. Exemplary cycloalkyl rings include cyclopropyl, cyclohexyl, and norbornene. Cycloalkyl groups may optionally be substituted with one or more substituents (such as those described herein).

[0079] "Heteroaryl" or "heterocyclic" refers to an aromatic monocyclic structure, preferably a 5- or 6-membered ring, whose ring structure contains at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The term "heteroaryl" or "heterocyclic" also includes polycyclic systems having two or more rings, wherein two or more atoms are shared by two adjacent rings, wherein at least one of the rings is heteroaromatic; for example, the other rings may be aromatic or non-aromatic carbocyclic rings, or heterocyclic rings. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine. Heteroaryl groups may optionally be substituted with one or more substituents (such as those described herein).

[0080] "Halogen" indicates fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

[0081] The term "optionally" means that the event or situation described below may or may not occur, and the description includes both the possibility that the event or situation may or may not occur, and the description includes both the possibility that the event or situation may occur and the possibility that it does not occur.

[0082] In some implementations, "replaced by one or more groups" means that one, two, three, or four hydrogen atoms of a specified atom or group are replaced by the same or different groups selected from a specified range of groups.

[0083] Unless otherwise indicated, the structural formulas described in this invention include all isomers (e.g., enantiomers, diastereomers, and geometric isomers (or conformational isomers)): for example, R and S configurations containing an asymmetric center, (Z) and (E) isomers of double bonds, and (Z) and (E) conformational isomers. Therefore, any single stereochemical isomer of the compounds of this invention, or its enantiomers, diastereomers, or mixtures of geometric isomers (or conformational isomers), is within the scope of this invention.

[0084] The definitions and conventions of stereochemistry used in this invention are generally referenced in the following literature: S.P. Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984); McGraw-Hill Book Company, New York; and Eliel J. E. and Wilen J. S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of this invention may contain asymmetric or chiral centers, and therefore exist as different stereoisomers. All stereoisomers of the compounds of this invention, including but not limited to diastereomers, enantiomers, stenotic isomers, and mixtures thereof, such as racemic mixtures, constitute a part of this invention. Many organic compounds exist in optically active forms, i.e., they are capable of rotating the plane of plane-polarized light. In describing optically active compounds, the prefixes D, L or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes d, l or (+), (i) are used for naming. The symbol for the rotation of plane-polarized light of a compound, ∠ or 1, indicates that the compound is levorotatory, while the prefix ⑴ or d indicates that the compound is dextrorotatory. These stereoisomers have the same chemical structure, but their stereostructures differ. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomeric mixtures. A 50:50 enantiomeric mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection during chemical reactions. The terms "racemic mixture" and "racemate" refer to a mixture of two equimolar enantiomers that lacks optical activity.

[0085] "Stereoisomers" are compounds that have the same chemical structure but whose atoms or groups are arranged differently in space. Stereoisomers include enantiomers, diastereomers, conformational isomers (rotational isomers), geometric isomers (cis / trans) isomers, and hindered isomers, etc.

[0086] The terms "tautomer" or "tautomerism form" refer to isomers of different energies that can interconvert through a low-energy barrier. For example, proton tautomers (i.e., proton-transfer tautomers) include interconversions via proton transfer, such as isomerization between keto-enol and imine-enamine forms. Valence tautomers include interconversions involving the recombination of bonding electrons.

[0087] "Chirality" refers to molecules that have the property that they cannot be superimposed on their mirror image; while "chirality" refers to molecules that can be superimposed on their mirror image.

[0088] "Enantiomers" refer to two non-overlapping but mirror-image isomers of a compound.

[0089] A diastereomer is a stereoisomer that has two or more chiral centers and whose molecules are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and reactivity. Mixtures of diastereomers can be separated by high-resolution analytical procedures such as electrophoresis and chromatography, for example, HPLC.

[0090] In some embodiments, the compositions of this disclosure may comprise two or more enantiomers or diastereomers of the compound, wherein a single enantiomer or diastereomer accounts for at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 98% by weight, or at least about 99% by weight or more of the total weight of all stereoisomers. Methods for producing substantially pure enantiomers are well known to those skilled in the art.

[0091] In addition, heavier isotopes, especially deuterium (i.e., 2 Substitution with H or D can provide certain therapeutic advantages. These advantages result from increased metabolic stability, such as increased in vivo half-life, reduced dose requirement, or improved therapeutic index. It should be understood that deuterium in this invention is considered a suitable substituent for compounds of formulas (I), (II), (III), (IV), and (V).

[0092] Unless otherwise stated, the compounds described herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds where hydrogen is replaced by deuterium or tritium, or where carbon is enriched. 13 C or 14Compounds having the structure of this invention, other than carbon substitution of C, are within the scope of this disclosure. The compounds of this disclosure optionally contain atomic isotopes in non-natural proportions at one or more atoms constituting such compounds. For example, the compounds may use isotopes such as, for example, deuterium (₂H), tritium (₃H), etc. 3 H), iodine 125 ( 125 I) or carbon 14 ( 14 C) Mark. Use 2 H, 11 C 13 C 14 C 15 C 12 N、 13 N、 15 N、 16 N、 16 O、 17 O、 14 F, 15 F, 16 F, 17 F, 18 F, 33 S, 34 S, 35 S, 36 S, 35 Cl、 37 Cl、 79 Br、 81 Br and 125 Isotopic substitutions of I are all considered. All isotopic variants of the compounds of this invention, whether or not they are radioactive, are covered within the scope of this invention.

[0093] "Pharmaceutically acceptable salts" refer to those salts that retain the bioavailability and properties of the parent compound. These salts include:

[0094] (1) It forms salts with acids, which are obtained by reacting the free base of the parent compound with inorganic or organic acids. Inorganic acids include hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, metaphosphoric acid, sulfuric acid, sulfurous acid and perchloric acid, etc. Organic acids include acetic acid, propionic acid, acrylic acid, oxalic acid, (D) or (L) malic acid, fumaric acid, maleic acid, hydroxybenzoic acid, γ-hydroxybutyric acid, methoxybenzoic acid, phthalic acid, methanesulfonic acid, ethanesulfonic acid, naphthalene-1-sulfonic acid, naphthalene-2-sulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, lactic acid, mandelic acid, succinic acid or malonic acid, etc.

[0095] (2) Salts formed by replacing acidic protons in the parent compound with metal ions or by coordination with organic bases. Examples of metals include alkali metal ions, alkaline earth metal ions, or aluminum ions. Examples of organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucosamine, etc.

[0096] "Pharmaceutical composition" refers to a mixture of one or more of the compounds of this invention, or their pharmaceutically acceptable salts, solvates, hydrates, or prodrugs, with other chemical components, such as a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate the administration of the drug to animals.

[0097] "Solvate" refers to an association formed by one or more solvent molecules with the compounds of this invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. The term "hydrate" refers to an association formed when the solvent molecules are water.

[0098] "Pharmaceutical carrier" refers to an inactive ingredient in a pharmaceutical composition that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the given compound. Examples include, but are not limited to: calcium carbonate, calcium phosphate, various sugars (e.g., lactose, mannitol, etc.), starch, cyclodextrin, magnesium stearate, cellulose, magnesium carbonate, acrylic polymers or methacrylic polymers, gels, water, polyethylene glycol, propylene glycol, ethylene glycol, castor oil or hydrogenated castor oil or polyethoxylated hydrogenated castor oil, sesame oil, corn oil, peanut oil, etc.

[0099] In addition to pharmaceutically acceptable carriers, the aforementioned pharmaceutical compositions may also include pharmaceutically commonly used excipients, such as antibacterial agents, antifungal agents, antimicrobial agents, preservatives, colorants, solubilizers, thickeners, surfactants, complexing agents, proteins, amino acids, fats, sugars, vitamins, minerals, trace elements, sweeteners, pigments, flavorings, or combinations thereof. Specific Implementation

[0101] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0102] The structure of the compounds was determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR measurements were performed using a Bruker Avance III 400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD) as solvents, and tetramethylsilane (TMS) as the internal standard. LC-MS was performed using a Shimadzu LC-MS2020 (ESI) system. HPLC was performed using a Shimadzu LC-20A system. MPLC (medium-pressure preparative chromatography) was performed using a Gilson GX-281 reversed-phase preparative chromatograph. Thin-layer chromatography (TLC) used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, with a thickness of 0.4 mm to 0.5 mm for product separation and purification. Silica gel column chromatography generally used Yantai Huanghai 200–300 mesh silica gel as the support. Chiral separation of SFC was performed using a Shimadzu LC-30AD SFC system.

[0103] In the examples, commonly used organic solvents are referred to using abbreviations well known in the art, such as: DCM refers to dichloromethane; THF refers to tetrahydrofuran; Dioxane refers to 1,4-dioxane; DMF refers to N,N-dimethylformamide; MeOH refers to methanol; EtOH refers to ethanol; HOAc refers to acetic acid, etc.

[0104] Example 1: (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (intermediate A)

[0105]

[0106] Step 1: (Z)-4-(1-(3-fluorophenyl)ethylene)-2-phenyloxazol-5(4H)-one

[0107] TiCl4 (160.00 g, 825.00 mmol) was added dropwise to 1.5 L of THF at -10 °C and stirred for 20 minutes. Then, a solution of 1-(3-fluorophenyl)ethane-1-one (b) (77.90 g, 563.00 mmol) in 200 mL of THF, 2-phenyloxazol-5(4H)-one (a) (100.00 g, 621.00 mmol), and pyridine (89.30 g, 1133.00 mol) were added dropwise, and the mixture was brought to room temperature and the reaction was terminated after 16 hours. The reaction solution was diluted with water, extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was slurried with MeOH (400 mL) at room temperature for 1 hour and filtered. The filter cake was purified by recrystallization from ethyl acetate at 60 °C to give the target compound (Z)-4-(1-(3-fluorophenyl)ethylene)-2-phenyloxazol-5(4H)-one (c). LCMS(ESI)calcd for C 17 H 12 FNO2282.09[M+H] + m / z,found282.2.

[0108] Step 2: (Z)-2-benzamido-3-(3-fluorophenyl)but-2-enoic acid methyl ester

[0109] A methanol (420 mL) solution of (Z)-4-(1-(3-fluorophenyl)ethylene)-2-phenyloxazol-5(4H)-one (c) (65.00 g, 231.00 mmol) was added to a methanol (30 mL) solution of NaOMe (0.62 g, 11.50 mmol) at 0 °C. The mixture was stirred at 0 °C for 0.5 h to terminate the reaction. The reaction solution was concentrated, diluted with water, extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give the target compound (Z)-2-benzamido-3-(3-fluorophenyl)but-2-enoic acid methyl ester (d). LCMS(ESI)calcd for C 18 H 16 FNO3314.11[M+H] + m / z, found 314.2.

[0110] Step 3: Methyl (2R,3S)-2-benzylamino-3-(3-fluorophenyl)butyrate

[0111] Pd / C (20.00 g, 55.38 mmol) was added to methyl (2R, 3S)-2-benzamido-3-(3-fluorophenyl)but-2-enoate (d) (70.00 g, 223.00 mmol) in 650 mL of MeOH at room temperature. The reaction was terminated at 50 °C for 16 hours under a hydrogen atmosphere. After cooling to room temperature, the product was filtered and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 30:1) and then chirally resolved by SFC to obtain the target compound (2R, 3S)-2-benzylamino-3-(3-fluorophenyl)butyrate methyl ester (e). LCMS (ESI) calcd for C 18 H 18 FNO3316.13[M+H] + m / z, found 316.2. 1 H NMR(400MHz, DMSO-d6)δ8.81(d,J=8.2Hz,1H),7.77(d,J=8.1Hz,2H),7.58–7.40(m,3H),7. 33–6.94(m,4H),4.75–4.65(m,1H),4.19–4.17(m,1H),3.40(s,3H),1.40(d,J=10.2Hz,3H).

[0112] Step 4: (2R,3S)-2-amino-3-(3-fluorophenyl)butyric acid

[0113] Acetic acid (500 mL) was added to a dilute hydrochloric acid (6 M, 500 mL) solution of methyl (2R,3S)-2-benzylamino-3-(3-fluorophenyl)butyrate (e) (35.00 g, 107.80 mmol) at room temperature. The reaction was terminated by heating to 120 °C for 16 hours. The reaction solution was cooled to room temperature and concentrated under reduced pressure. The crude product was washed with DCM (300 mL * 2). The filter cake was filtered and dried to obtain the target compound (2R,3S)-2-amino-3-(3-fluorophenyl)butyric acid (f). LCMS (ESI) calcd for C 10 H 12 FNO2198.09[M+H] + m / z,found198.2. 1 H NMR (400MHz, DMSO) δ8.60–8.55(m,3H),7.44–7.31(m,1H),7.22–7.06(m,3H),4.10–3.97(m,1H),3.50–3.38(m,1H),1.37(dd,J=18.6,7.2Hz,3H).

[0114] Step 5: (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid

[0115] HNO3 (12.19 g, 132.00 mmol, 68.0% purity) was slowly added dropwise to (2R,3S)-2-amino-3-(3-fluorophenyl)butyric acid (f) (25.00 g, 107.00 mmol) in 100 mL of concentrated sulfuric acid at -20 °C. The reaction was terminated at 0 °C for 0.5 h. The reaction solution was quenched in ice water (500 mL) and the pH was adjusted to 8. After filtration and concentration, the crude product was purified by reverse preparative chromatography to obtain the target compound (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (g).

[0116] LCMS(ESI)calcd for C 10 H 11 FN₂O₄243.07[M+H] + m / z, found 243.2.

[0117] Step 6: (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid

[0118] At room temperature, (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (g) (2.00 g, 8.20 mmol) and Boc₂O (3.58 g, 16.40 mmol) were added sequentially to THF (20 mL), and the reaction was terminated at room temperature for 1 hour. The reaction solution was diluted with water, extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the target compound (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid.

[0119] LCMS(ESI)calcd for C 15 H 19 FN₂O₆342.12[M+H] + m / z, found 343.2.

[0120] Example 2: N-((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 1)

[0121]

[0122] Step 1: ((2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-oxo-1-(prop-2-yn-1-ylamino)but-2-yl)carbamate tert-butyl ester

[0123] At room temperature, (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (intermediate A) (1.20 g, 3.50 mmol), 2-propane-1-amine (0.19 g, 3.50 mmol), HBTU (2.65 g, 7.00 mmol), and N,N-diisopropylethylamine (0.90 g, 7.00 mmol) were dissolved together in DMF (10 mL), and the reaction was terminated at room temperature for 1 hour. After the reaction, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to give the target compound ((2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-oxo-1-(propane-2-yn-1-ylamino)but-2-yl)carbamate tert-butyl ester (1a). LCMS(ESI)cal cdfor C 18 H 22 FN3O5[M+H] + m / z 380.2, found 380.1. 1 H NMR (400MHz, CDCl3) δ8.05–7.96(m,1H),7.22–7.14(m,2H),6.54–6.26(m,1H),5.35–5. 05(m,1H),4.38–4.25(m,1H),4.17–3.67(m,3H),3.48–3.27(m,1H),1.43–1.33(m,12H).

[0124] Step 2: ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)tert-butyl carbamate (1b)

[0125] Under an oxygen atmosphere at room temperature, tert-butyl carbamate ((2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-oxo-1-(prop-2-yn-1-ylamino)but-2-yl)carbamate (1a) (1.20 g, 3.20 mmol), PhI(OAc)2 (0.10 g, 0.03 mmol), and lithium iodide (0.04 g, 0.03 mmol) were dissolved in DCM (50 mL), and the reaction was terminated after 16 hours at room temperature under irradiation with a 40 W household fluorescent bulb. After the reaction, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain the target compound ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)carbamate (1b). LCMS(ESI)calcd for C 18 H 20 FN3O6[M+H] + m / z 394.1, found 394.2. 1 H NMR(400MHz, CDCl3)δ9.83–9.66(m,1H),8.00(t,J=5.3Hz,1H),7.74(s,1H),7.16–7.04 (m,2H),5.43–5.10(m,2H),3.53(t,J=9.4Hz,1H),1.43–1.37(m,9H),1.34–1.25(m,3H).

[0126] Step 3: ((1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate (1c)

[0127] At room temperature, tert-butyl carbamate (1b) (0.65 g, 1.65 mmol), dimethylamine hydrochloride (0.13 g, 1.65 mmol), and sodium cyanoborohydride (0.01 g, 0.16 mmol) were dissolved in methanol (10 mL), and the reaction was terminated at room temperature after 16 hours. After the reaction, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to give the target compound (1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)carbamate (1c). LCMS(ESI)calcd for C 20 H 27FN4O5[M+H] + m / z 423.2, found 423.1.

[0128] Step 4: (1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)prop-1-amine (1d)

[0129] At room temperature, tert-butyl carbamate (1c) (0.29 g, 0.68 mmol) was dissolved in ethyl acetate solution of HCl (1.0 M, 10 mL), and the reaction was terminated after 16 hours at room temperature. After the reaction was completed, the target compound (1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)prop-1-amine (1d) was obtained by concentration. LCMS(ESI)calcd for C 15 H 19 FN4O3[M+H] + m / z 323.1, found 323.1.

[0130] Step 5: N-((1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1e)

[0131] At room temperature, (1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl-1-amine (1d) (0.20 g, 0.62 mmol), propionic anhydride (0.08 g, 0.62 mmol), and N,N-diisopropylethylamine (0.16 g, 1.24 mmol) were dissolved together in DMF (10 mL), and the reaction was terminated at room temperature for 5 hours. After the reaction, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give the target compound N-((1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1e). LCMS (ESI) calcd for C 18 H 23 FN4O4[M+H] + m / z 379.2, found 379.1.

[0132] Step 6: N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1f)

[0133] At room temperature, N-((1R,2S)-1-(5-((dimethylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1e) (0.50 g, 1.32 mmol) and Pd / C (0.05 g, 10%) were dissolved together in a mixed solution of ethanol (10 mL) and tetrahydrofuran (10 mL), and the reaction was terminated at room temperature under a hydrogen atmosphere for 16 hours. After the reaction was completed, the solution was filtered and concentrated to obtain the target compound N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1f). LC MS (ESI) calcd for C 18 H 25 FN4O2[M+H] + m / z 349.2, found 349.2.

[0134] Step 7: Tert-butyl((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl))-2-fluorophenyl)amino)-2-oxoethyl)carbamate (1g)

[0135] At room temperature, N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1f) (0.43 g, 1.23 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetic acid (0.33 g, 1.23 mmol), HATU (0.94 g, 2.46 mmol), and N,N-diisopropylethylamine (0.32 g, 2.46 mmol) were dissolved together in DMF (5 mL), and the reaction was terminated at 50 °C for 16 hours. After the reaction was completed, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase column chromatography (acetonitrile:water = 1:1) to give the target compound tert-butyl((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl))-2-fluorophenyl)amino)-2-oxoethyl)carbamate (1 g). LCMS (ESI) calcd for C 32 H 48 FN5O5[M+H] + m / z 602.4, found 602.3.

[0136] Step 8: N-((1R,2S)-2-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-1-(5-((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1h)

[0137] At room temperature, tert-butyl((S)-1-cycloheptayl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propanoylaminopropane-2-yl))-2-fluorophenyl)amino)-2-oxoethyl)carbamate (1 g) (0.10 g, 0.16 mmol) was dissolved in dichloromethane (5 mL), and TFA (5 mL) was added dropwise. The reaction was terminated at room temperature for 1 hour. After the reaction was completed, the solution was concentrated to obtain N-((1R,2S)-2-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1 h). LCMS(ESI)calcd for C 27 H 40 FN5O3[M+H] + m / z502.3, found 502.3.

[0138] Step 9: N-((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 1)

[0139] At room temperature, N-((1R,2S)-2-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)propyl)propionamide (1h) (0.08 g, 0.16 mmol), HBTU (0.12 g, 0.32 mmol), and N,N-diisopropylethylamine (0.04 g, 0.32 mmol) were dissolved together in DMF (2 mL), and the reaction was terminated at room temperature for 1 hour. After the reaction was completed, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, respectively. The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative HPLC column chromatography to obtain the target compound N-((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 1). LCMS(ESI)calcd for C 34 H48 FN7O4[M+H] + m / z 638.4, found 638.4. 1 H NMR (400MHz, DMSO) δ9.80(s,1H),8.51(d,J=8.8Hz,1H),8.42(d,J=8.4Hz,1H),7.64(t,J=8.0Hz,1H ),7.48(d,J=2.0Hz,1H),7.03(d,J=11.6Hz,1H),6.92–6.88(m,2H),6.82(s,1H),5.41–5.34(m,1H), 5.09(t,J=9.2Hz,1H),4.58(t,J=8.4Hz,1H),3.30–3.21(m,3H),2.20–2.05(m,4H),1.99(s,6H),1. 71–1.60(m,4H),1.56–1.44(m,4H),1.38–1.33(m,8H),1.25(d,J=6.8Hz,4H),0.98(t,J=7.6Hz,3H).

[0140] Example 3: N-((S)-1-cycloheptyl-2-((2-fluoro-4-((1R,2S)-1-(5-(piperidin-1-ylmethyl)oxazol-2-yl)-1-propionylaminopropane)-2-yl)phenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 2)

[0141]

[0142] Step 1: ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(piperidin-1-ylmethyl)oxazol-2-yl)propyl)tert-butyl carbamate (2a)

[0143] At room temperature, tert-butyl carbamate (1b) (0.72 g, 1.83 mmol), piperidine hydrochloride (223 mg, 1.83 mmol), and sodium cyanoborohydride (115 mg, 1.83 mmol) were dissolved in methanol (10 mL), and the reaction was terminated at room temperature after 16 hours. The reaction solution was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to give tert-butyl carbamate (2a).

[0144] 1 H NMR (400MHz, DMSO) δ8.01(t,J=8.2Hz,1H),7.75(d,J=8.8Hz,1H),7.46(d,J=12.7Hz,1H),7.24(d,J=8.4Hz,1H ),6.81(s,1H),4.89(t,J=9.0Hz,1H),3.50–3.36(m,3H),2.15(s,4H),1.43–1.34(m,13H),1.32–1.28(m,5H).

[0145] Steps 2 to 7 refer to the synthesis scheme of steps 4 to 9 in Example 2 to obtain the target compound N-((S)-1-cycloheptyl-2-((2-fluoro-4-((1R,2S)-1-(5-(piperidin-1-ylmethyl)oxazol-2-yl)-1-propionylaminopropane)-2-yl)phenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 2).

[0146] LCMS(ESI)calcd for C 37 H 53 FN7O4 + [M+H] + m / z 678.41, found 678.45.

[0147] 1 H NMR (400MHz, DMSO) δ9.79(s,1H),8.51(d,J=8.8Hz,1H),8.41(d,J=8.8Hz,1H),7.67(t,J=8.0 Hz,1H),7.48(s,1H),7.03(d,J=12.4Hz,1H),6.91–6.86(m,2H),6.77(s,1H),5.41–5.34(m,1H ),5.10(t,J=9.6Hz,1H),4.58(t,J=8.4Hz,1H),3.41–3.34(m,2H),3.27–3.18(m,2H),2.22–2 .09(m,7H),1.75–1.45(m,10H),1.38–1.32(m,11H),1.27–1.23(m,5H),0.98(t,J=7.6Hz,3H).

[0148] Example 4: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 6)

[0149]

[0150] Step 1: ((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate (6a)

[0151] Acetic acid (0.05 g, 0.76 mmol) and N-methylcyclopropylamine (0.05 g, 0.76 mmol) were dissolved in methanol (3 mL) at room temperature. Then, tert-butyl carbamate (1b) (0.27 g, 0.69 mmol) was added with stirring. The reaction mixture was then reacted at room temperature for 15 minutes, followed by the addition of sodium cyanoborohydride (0.12 g, 1.91 mmol). The reaction was terminated after 2 hours at room temperature. A saturated aqueous solution of sodium bicarbonate was slowly added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 20 / 1) to obtain the target compound ((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)carbamate tert-butyl ester (6a).

[0152] LC-MS(ESI)calcd for C 22 H 29 FN4O5[M+H] + m / z 449.2, found 449.4.

[0153] Step 2: N-((2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-yl)methyl)-N-methylcyclopropylamine (6b)

[0154] At room temperature, a solution of tert-butyl carbamate (6a) (0.26 g, 0.58 mmol) in dichloromethane (2 mL) was added to a solution of hydrogen chloride / 1,4-dioxane (4 M, 2 mL). The reaction was then terminated after 2 hours at room temperature. The reaction solution was concentrated to give crude N-((2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-yl)methyl)-N-methylcyclopropylamine (6b), which was used directly in the next step without further purification.

[0155] LC-MS(ESI)calcd for C 17 H 21 FN4O3[M+H] + m / z 349.2, found 349.3.

[0156] Step 3: N-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (6c)

[0157] N-((2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-yl)methyl)-N-methylcyclopropylamine (6b) (0.26 g, crude hydrochloride) and triethylamine (0.23 g, 2.24 mmol) were dispersed in dichloromethane (5 mL) at room temperature, and propionyl chloride (0.19 g, 1.49 mmol) was added dropwise with stirring. The reaction was terminated after 2 hours at room temperature. The reaction solution was extracted with dichloromethane after adding saturated sodium bicarbonate solution. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography (wet loading, eluent: dichloromethane / methanol = 20 / 1) to obtain the target compound N-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (6c). LC-MS (ESI) calcd for C 20 H 25 FN4O4[M+H] + m / z 405.2,fo und 405.0.

[0158] Step 4: N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)propyl)propionamide (6d)

[0159] N-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (6c) (0.23 g, 0.57 mmol), zinc powder (0.37 g, 5.68 mmol), and ammonium chloride (0.61 g, 11.37 mmol) were dispersed in tetrahydrofuran (3 mL) and water (1 mL) at room temperature. The reaction mixture was then heated to 80 °C for 5 hours to terminate the reaction. A saturated aqueous solution of potassium carbonate was added to the cooled reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 10 / 1) to give N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)propyl)propionamide (6d). LC-MS (ESI) calcd for C 20 H 27 FN4O2[M+H] + m / z375.2,fou nd 375.4.

[0160] Step 5: ((S)-1,1-Dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)tert-butyl carbamate (6e)

[0161] At room temperature, (S)-2-((tert-Butoxycarbonyl)amino)-3,3-dicyclopropylpropionic acid (0.20 g, 0.75 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.28 g, 0.75 mmol), and N,N-diisopropylethylamine (0.15 g, 1.12 mmol) were added to a solution of N,N-dimethylformamide (2 mL) containing N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)propyl)propionamide (6d) (0.14 g, 0.37 mmol) at room temperature. The reaction was terminated after microwave reaction at 30 °C for 16 hours. The reaction solution was filtered, and the filtrate was purified by reversed-phase C18 column chromatography (mobile phase: 0.1% formic acid aqueous solution / acetonitrile = 2 / 1) to give ((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)carbamate (6e). LC-MS (ESI) calculation for C 34 H48 FN5O5[M+H] + m / z 626.4, found 627.1; [M+Na] + m / z 648.4, found648.6.

[0162] Step 6: (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f)

[0163] A solution of hydrogen chloride / 1,4-dioxane (4M, 1 mL) was added to a stirred solution of ((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)carbamate (6e) (0.10 g, 0.16 mmol) in 1 mL of dichloromethane. The reaction was then stopped after stirring at room temperature for 2 hours. The reaction solution was concentrated to obtain crude (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f). The product was used directly in the next step without further purification. LC-MS (ESI) calcd for C 29 H 40 FN5O3[M+H] + m / z 526.3, found 526.8; [M+Na] + m / z 548.3, found 548.4.

[0164] Step 7: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 6)

[0165] At room temperature, 1-isopropyl-1H-pyrazole-5-carboxylic acid (0.07 g, 0.44 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.13 g, 0.36 mmol), and N,N-diisopropylethylamine (0.14 g, 1.07 mmol) were added to a solution of N,N-dimethylformamide (6f) containing (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f) (0.10 g, crude) (2 mL). The reaction was terminated by microwave reaction at 30 °C for 16 hours. After the reaction was completed, the system was filtered, and the filtrate was purified by high performance liquid chromatography (mobile phase: 10 mM ammonium bicarbonate aqueous solution / acetonitrile = 1 / 1). The obtained product components were freeze-dried to obtain N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 6).

[0166] LC-MS(ESI)calcd for C 36 H 48 FN7O4[M+H] + m / z 662.4, found 662.7.

[0167] 1H NMR (400MHz, DMSO-d6) δ9.82(s,1H),8.48(d,J=8.9Hz,1H),8.39(d,J=8.9Hz,1H),7.73(t,J=8.3Hz,1H),7.49(d,J=1.9Hz,1H),7 .05(dd,J=12.2,1.7Hz,1H),6.90(dd,J=9.2,6.0Hz,2H),6.81(s,1H),5.45-5.33(m,1H),5.12(t,J=9.2Hz,1H),4.92(t,J=8.3Hz, 1H),3.53(s,2H),3.28-3.19(m,1H),2.21-2.08(m,2H),2.05(s,3H),1.46-1.41(m,1H),1.36(dd,J=15.3,6.6Hz,6H),1.25(d,J= 7.0Hz, 3H), 0.97 (t, J = 7.6Hz, 3H), 0.90 (d, J = 8.2Hz, 1H), 0.82 (d, J = 5.3Hz, 1H), 0.72 (dd, J = 17.2, 9.3Hz, 1H), 0.40-0.10 (m, 12H).

[0168] Example 5: (S)-3,3-dicyclopropyl-2-(2,2-difluoro-2-(6-methoxypyridin-3-yl)acetamido)-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)(Compound 7)

[0169]

[0170] Step 1: ((1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate (7a)

[0171] 3,3-Difluoropyrrolidine hydrochloride (0.73 g, 5.08 mmol) was dissolved in anhydrous methanol (6 mL) at room temperature. Triethylamine (1.03 g, 10.17 mmol) was added with stirring, and the reaction was allowed to proceed for 2 minutes at room temperature. Glacial acetic acid (0.61 g, 10.17 mmol) and ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)carbamate tert-butyl ester (1b) (1.00 g, 2.54 mmol) were then added sequentially with stirring. The reaction mixture was subsequently heated to 30 °C and reacted for 1 hour. Sodium cyanoborohydride (0.32 g, 5.08 mmol) was then added, and the reaction was terminated after 1 hour at room temperature. The reaction mixture was extracted with ethyl acetate after adding saturated sodium bicarbonate solution. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 50 / 1) to obtain the target compound ((1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)carbamate tert-butyl ester (7a).

[0172] LC-MS(ESI)calcd for C 22 H 27 F3N4O5[M+H] + m / z 485.2, found 485.2.

[0173] Steps 2 to 6 were performed according to the synthesis scheme of steps 2 to 6 in Example 4, yielding the target compound (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propamido (7f).

[0174] LC-MS(ESI)calcd for C 29 H 38 F3N5O3[M+H] + m / z 562.3, found 562.4.

[0175] Step 7: (S)-3,3-dicyclopropyl-2-(2,2-difluoro-2-(6-methoxypyridin-3-yl)acetamido)-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (Compound 7)

[0176] At room temperature, 2,2-difluoro-2-(6-methoxypyridin-3-yl)acetic acid (0.05 g, 0.24 mmol) and (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (7f) (0.07 g, crude) were dissolved in N,N-dimethylformamide (3 mL). 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.09 g, 0.24 mmol) and N,N-diisopropylethylamine (0.09 g, 0.7 mmol) were added sequentially with stirring. The reaction mixture was microwaved at 30 °C for 16 hours. The reaction solution was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile). The product components were freeze-dried to obtain the target compound (S)-3,3-dicyclopropyl-2-(2,2-difluoro-2-(6-methoxypyridin-3-yl)acetamido)-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (compound 7).

[0177] LC-MS(ESI)calcd for C 37 H 43 F5N6O5[M+H] + m / z 747.3, found 747.6.

[0178] 1HNMR(400MHz,DMSO-d6)δ9.92(s,1H),8.93(d,J=8.8Hz,1H),8.49(d,J=8.9Hz,1H),8.43(s,1H),7.92(dd,J=8.9,2.4Hz,1 H),7.70(t,J=8.3Hz,1H),7.06(d,J=11.2Hz,1H),6.95(d,J=8.6Hz,1H),6.90(d,J=8.2Hz,1H),6.84(s,1H),5.12(t,J=9. 2Hz,1H),4.92-4.78(m,1H),3.89(s,3H),3.62-3.53(m,2H),3.31-3.19(m,3H),2.80(t,J=13.3Hz,2H),2.24-2.08(m,4H) ,1.25(d,J=6.9Hz,3H),0.97(t,J=7.6Hz,3H),0.80-0.66(m,3H),0.43-0.34(m,1H),0.32-0.24(m,1H),0.23-0.04(m,6H).

[0179] Example 6: N-((S)-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-(4,4-difluorocyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 8)

[0180]

[0181] Step 1: ((S)-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-(4,4-difluorocyclohexyl)-2-oxoethyl)tert-butyl carbamate (8a)

[0182] At room temperature, N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)propyl)propionamide (6d) (0.05 g, 0.13 mmol) and (S)-2-((tert-butyloxycarbonyl)amino)-2-(4,4-difluorocyclohexyl)acetic acid (0.08 g, 0.27 mmol) were dissolved in micronized N,N-dimethylformamide (3 mL). 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.10 g, 0.27 mmol) and N,N-diisopropylethylamine (0.10 g, 0.80 mmol) were added sequentially with stirring. The reaction mixture was microwaved at 30 °C for 16 hours, after which the reaction was terminated. The reaction solution was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 40 / 60) to obtain the target compound ((S)-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-(4,4-difluorocyclohexyl)-2-oxoethyl)carbamate tert-butyl carbamate (8a).

[0183] LCMS(ESI)calcd for C 33 H 46 F3N5O5[M+H] + m / z 650.4, found 650.4.

[0184] Steps 2 and 3 refer to steps 6 and 7 of Example 4 to obtain the target compound N-((S)-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-(4,4-difluorocyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 8).

[0185] LCMS(ESI)calcd for C 35 H 46 F3N7O4[M+H] + m / z 686.4, found 686.4.

[0186] 1H NMR (400MHz, DMSO-d6) δ9.88(s,1H),8.55(d,J=8.3Hz,1H),8.49(d,J=8.8Hz,1H),7.69(t,J=8.3Hz,1H),7.49(d,J=1.8Hz ,1H),7.05(d,J=12.2Hz,1H),6.95-6.89(m,2H),6.81(s,1H),5.45-5.33(m,1H),5.11(t,J=9.2Hz,1H),4.63(t,J=8.5Hz, 1H),3.53(s,2H),3.27-3.20(m,1H),2.17-2.09(m,3H),2.06-1.97(m,5H),1.90-1.77(m,2H),1.76-1.65(m,2H),1.47-1. 39(m,2H),1.35(dd,J=9.0,6.7Hz,7H),1.25(d,J=7.0Hz,3H),0.98(t,J=7.6Hz,3H),0.37-0.29(m,2H),0.20-0.13(m,2H).

[0187] Example 7: N-((S)-1-(4,4-difluorocyclohexyl)-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (Compound 9)

[0188]

[0189] Steps 1 to 3 were performed according to the synthesis scheme of Steps 1 to 3 in Example 6, yielding the target compound N-((S)-1-(4,4-difluorocyclohexyl)-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (compound 9).

[0190] LC-MS(ESI)calcd for C 34 H 42 F5N7O4[M+H] + m / z 708.3, found 708.7.

[0191] 1H NMR(400MHz, DMSO-d6)δ9.90(s,1H),8.56(d,J=8.2Hz,1H),8.50(d,J=8.8Hz,1H),7.67(t,J=8.3Hz,1H),7.47(d,J=2.0Hz,1H),7 .07(d,J=12.1Hz,1H),6.99(d,J=2.0Hz,1H),6.90(d,J=8.3Hz,1H),6.84(s,1H),5.13(t,J=9.2Hz,1H),4.64(t,J=8.4Hz,1H),4. 46(q,J=7.2Hz,2H),3.63-3.52(m,2H),3.28-3.21(m,1H),2.79(t,J=13.5Hz,2H),2.70-2.62(m,1H),2.48-2.41(m,1H),2.22-2. 10(m,4H),2.09-1.94(m,3H),1.91-1.66(m,4H),1.52-1.42(m,1H),1.37-1.32(m,1H),1.29-1.23(m,6H),0.98(t,J=7.6Hz,3H).

[0192] Example 8: N-((1S)-2-((4-((1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 10)

[0193]

[0194] Step 1: tert-butyl((1S)-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propionylaminopropane)-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate (10a)

[0195] At room temperature, N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)propyl)propionamide (7d) (0.41 g, 1.00 mmol), (2S)-2-((tert-butoxycarbonyl)amino)-2-((1R,4S)-4-methylcyclohexyl)acetic acid (0.27 g, 1.00 mmol), HATU (0.76 g, 2.00 mmol), and N,N-diisopropylethylamine (0.26 g, 2.00 mmol) were dissolved together in DMF (5 mL), and the reaction was terminated at 50 °C for 16 hours. After the reaction was completed, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by reverse-phase column chromatography (acetonitrile:water = 1:1) to obtain the target compound tert-butyl((1S)-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propionylaminopropane)-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate (10a).

[0196] LCMS(ESI)calcd for C 34 H 48 F3N5O5[M+H] + m / z 663.0, found 664.1.

[0197] Steps 2 and 3 refer to steps 6 and 7 of Example 4 to obtain the target compound N-((1S)-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 10).

[0198] LCMS(ESI)calcd for C 36 H 48 F3N7O4[M+H] + m / z 699.8, found 701.4.

[0199] 1H NMR (400MHz, DMSO-d6) δ9.79(s,1H),8.50(d,J=8.8Hz,1H),8.41(d,J=8.4Hz,1H),7.66(t,J=8.4Hz,1H),7.48(d,J=2.0Hz,1H ),7.05(d,J=12.0Hz,1H),6.90(dd,J=10.4,5.1Hz,2H),6.84(s,1H),5.37(dd,J=13.2,6.8Hz,1H),5.12(t,J=9.2Hz,1H),4.4 9(t,J=8.0Hz,1H),3.58(d,J=5.6Hz,2H),3.30–3.21(m,2H),2.79(s,2H),2.50-2.43(m,4H),2.15(dd,J=12.4,7.6Hz,4H),1. 79(s,2H),1.70-1.60(m,3H),1.35(dd,J=9.2,6.8Hz,6H),1.25(d,J=6.8Hz,3H),0.98(t,J=7.6Hz,3H),0.85(d,J=6.4Hz,5H).

[0200] Example 9: N-((1S)-2-((4-((1R,2S)-1-(5-(((cyclopropylamino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl))amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 11)

[0201]

[0202] Step 1: ((1R,2S)-1-(5-((cyclopropylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate

[0203] At room temperature, tert-butyl carbamate (1b) (0.95 g, 2.41 mmol), cyclopropylamine hydrochloride (0.22 g, 2.41 mmol), and sodium cyanoborohydride (0.08 g, 1.20 mmol) were dissolved in methanol (20 mL), and the reaction was terminated at room temperature after 16 hours. After the reaction, the mixture was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to give the target compound (1R,2S)-1-(5-((cyclopropylamino)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)carbamate (11a).

[0204] LCMS(ESI)calcd for C 21 H 27 FN4O5[M+H] + m / z 434.1, found 435.2.

[0205] Step 2: ((2-((1R,2S)-1-((tert-butoxycarbonyl)amino)-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-yl)methyl)(cyclopropyl)carbamate (11b)

[0206] At room temperature, 0.76 g (1.75 mmol) of tert-butyl carbamate (11a) was dissolved in chloroform (30 mL) and saturated sodium carbonate solution (10 mL), and the reaction was terminated at room temperature after 3 hours. After the reaction was completed, the target compound (2-((1R,2S)-1-((tert-butoxycarbonyl)amino)-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-yl)methyl)(cyclopropyl)carbamate (11b) was obtained by concentration and purification with ethyl acetate.

[0207] LCMS(ESI)calcd for C 29 H 33 FN4O7[M+H] + m / z 569.0, found 569.9.

[0208] Steps 3 to 5 refer to the synthesis scheme of steps 2 to 4 in Example 4.

[0209] Steps 6 to 8 refer to steps 1 to 3 of Example 8 to obtain the target compound benzylcyclopropyl ((2-((1R,2S)-2-(3-fluoro-4-((2S)-2-(1-isopropyl-1H-pyrazole-5-carbamate))-2-((1R,4S)-4-methylcyclohexyl)acetamido)phenyl)-1-propionylaminopropyl)oxazol-5-yl)methyl)carbamate (11h)

[0210] LCMS(ESI)calcd for C 43 H 54 FN7O6[M+Na] + m / z 783.1, found 784.2.

[0211] Step 9: N-((1S)-2-((4-((1R,2S)-1-(5-(((cyclopropylamino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl))amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 11)

[0212] At room temperature, benzylcyclopropyl ((2-((1R,2S)-2-(3-fluoro-4-((2S)-2-(1-isopropyl-1H-pyrazole-5-carbamate))-2-((1R,4S)-4-methylcyclohexyl)acetamido)phenyl)-1-propionylaminopropyl)oxazol-5-yl)methyl)carbamate (11h) (0.06g, 0.08mmol), Pd / C (20mg, 5%) were dissolved together in ethanol / tetrahydrofuran (20mL), and the reaction was terminated after 16 hours at room temperature under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated, and then separated by high performance liquid preparative chromatography (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound N-((1S)-2-((4-((1R,2S)-1-(5-(((cyclopropylamino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl))amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 11).

[0213] LCMS(ESI)calcd for C 35 H 48 FN7O4[M+H] + m / z 650.8, found 650.5.

[0214] 1H NMR (400MHz, DMSO) δ9.78(s,1H),8.43(dd,J=12.4,8.8Hz,2H),7.65(t,J=8.4Hz,1H),7.48(d,J=1.6Hz,1H),7.05(d,J=12. 0Hz,1H),6.93–6.88(m,2H),6.76(s,1H),5.38(dt,J=13.2,6.4Hz,1H),5.13(t,J=9.2Hz,1H),4.49(t,J=8.4Hz,1H),3.60( s,2H),3.29–3.24(m,1H),2.22-2.09(m,2H),1.85-1.79(m,3H),1.70-1.60(m,3H),1.35(dd,J=8.8,6.8Hz,6H),1.24(d,J= 6.8Hz, 6H), 1.06 (d, J = 12.0Hz, 1H), 0.98 (t, J = 7.6Hz, 3H), 0.85 (d, J = 6.4Hz, 5H), 0.26 (q, J = 6.8Hz, 2H), 0.16–0.09 (m, 2H).

[0215] Example 10: N-[(1R,2S)-2-{3-fluoro-4-[(2S)-2-[(2-isopropylpyrazole-3-yl)carbamoyl]-2-((1R,4S)-4-methylcyclohexyl)acetamyl]phenyl}-1-[5-(pyrrolidone-1-ylmethyl)-1,3-oxazol-2-yl]propyl]propionamide (Compound 12)

[0216]

[0217] Step 1: N-[(1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-[5-(pyrrolidone-1-ylmethyl)-1,3-oxazol-2-yl]propyl] tert-butyl carbamate (12a)

[0218] At room temperature, N-[(1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyl-1,3-oxazol-2-yl)propyl] tert-butyl carbamate (1b) (1.00 g, 2.50 mmol) and pyrrolidine hydrochloride (0.27 g, 2.50 mmol) were dissolved in MeOH (10 mL), and a mixture of NaBH3CN (0.16 g, 2.50 mmol) was added. The reaction was terminated at room temperature for 4 hours. The reaction solution was concentrated. The crude product was purified by silica gel column chromatography (DCM / MeOH = 98:2) to give N-[(1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-[5-(pyrrolidine-1-ylmethyl)-1,3-oxazol-2-yl]propyl] tert-butyl carbamate (12a).

[0219] 1 H NMR (400MHz, CDCl3) δ7.97(t,J=8.4Hz,1H),7.09(t,J=10.0Hz,2H),6.93(s,1H),5.39(d,J=9.2Hz,1H),5.11(t,J=7 .6Hz,1H),3.79(q,J=14.4Hz,2H),3.56–3.38(m,1H),2.68(s,4H),1.88(s,4H),1.42(s,9H),1.36(d,J=7.2Hz,3H).

[0220] Steps 2 to 7 refer to the synthesis scheme of steps 3 to 8 in Example 9 to obtain N-[(1R,2S)-2-{3-fluoro-4-[(2S)-2-[(2-isopropylpyrazole-3-yl)carbamoyl]-2-((1R,4S)-4-methylcyclohexyl)acetamyl]phenyl}-1-[5-(pyrrolidone-1-ylmethyl)-1,3-oxazol-2-yl]propyl]propionamide (compound 12).

[0221] LCMS(ESI)calcd for C 36 H 50 FN7O4[M+H] + m / z 664.4, found 664.8.

[0222] 1 H NMR (400MHz, DMSO) δ9.79(s,1H),8.50(d,J=8.8Hz,1H),8.42(d,J=8.0Hz,1H),7.68(t,J=8.4Hz,1H),7.48(d,J=2.0H z,1H),7.03(d,J=12.0Hz,1H),6.92(d,J=2.0Hz,1H),6.88(d,J=8.4Hz,1H),6.78(s,1H),5.38(dt,J=13.2,6.4Hz,1H) ,5.11(t,J=9.2Hz,1H),4.50(t,J=8.4Hz,1H),3.49(q,J=14.4Hz,2H),3.26–3.21(m,1H),2.26(s,4H),2.13–2.09(m,2 H),1.81–1.53(m,9H),1.36–1.31(m,6H),1.25–1.19(m,5H),1.07–1.00(m,1H),0.99–0.96(m,3H),0.87–0.84(m,5H).

[0223] Example 11: N-((1S)-2-((4-((1R,2S)-1-(5-(azacyclobutane-1-ylmethyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 13)

[0224]

[0225] Step 1: ((1R,2S)-1-(5-(azacyclobutane-1-ylmethyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate (13a)

[0226] At room temperature, tert-butyl carbamate (1b) (1.00 g, 2.54 mmol), cyclobutamine hydrochloride (0.24 g, 2.54 mmol), and sodium cyanoborohydride (0.39 g, 2.54 mmol) were dissolved together in methanol (20 mL), and the reaction was terminated at room temperature after 16 hours. After the reaction, the product was extracted with ethyl acetate, and the organic layer was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by silica gel column chromatography (dichloromethane:methanol = 20:1) to give tert-butyl carbamate (13a).

[0227] LCMS(ESI)calcd for C 21 H 27 FN4O5[M+H] + m / z 435.2, found 435.2.

[0228] Steps 2 to 7 were performed following the synthetic scheme of steps 3 to 8 in Example 9 to obtain the target compound N-((1S)-2-((4-((1R,2S)-1-(5-(azacyclobutane-1-ylmethyl)oxazol-2-yl)-1-propionylaminopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 13).

[0229] LCMS(ESI)calcd for C 35 H 48 FN7O4[M+H] + m / z 650.3, found 650.5.

[0230] 1H NMR (400MHz, DMSO) δ9.81(s,1H),8.51–8.43(m,2H),7.66(t,J=8.3Hz,1H),7.48(s,1H),7.03(d,J=11.4H z,1H),6.93–6.88(m,2H),6.76(s,1H),5.41–5.34(m,1H),5.10(t,J=9.1Hz,1H),4.49(t,J=8.1Hz,1H),3 .38-3.32(m,2H),3.26-3.24(m,1H),3.00(t,J=6.8Hz,3H),2.27–2.09(m,4H),2.04–1.96(m,2H),1.85-1 .78(m,4H),1.66-1.61(m,4H),1.37–1.33(m,6H),1.23(s,3H),0.99–0.95(m,3H),0.85(d,J=6.3Hz,4H).

[0231] Example 12: N-((1S)-2-((4-((1R,2S)-1-(5-((3,3-difluoroazacyclobutane-1-yl)methyl)oxazol-2-yl)-1-propamidopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 14)

[0232]

[0233] Step 1: ((1R,2S)-1-(5-((3,3-difluoroazacyclobutane-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)tert-butyl carbamate

[0234] At room temperature, tert-butyl carbamate (1b) (1.00 g, 2.54 mmol), 3,3-difluoroazacyclobutane hydrochloride (0.33 g, 2.54 mmol), and sodium cyanoborohydride (0.16 g, 2.54 mmol) were dissolved in methanol (20 mL), and the reaction was terminated at room temperature after 16 hours. After the reaction was completed, the crude product was concentrated and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain the target compound (14a) tert-butyl carbamate (1b).

[0235] LCMS(ESI)calcd for C 21 H 25 F3N4O5[M+H] + m / z 471.18, found 471.2.

[0236] Steps 2 to 7 are performed following the synthetic scheme of steps 3 to 8 in Example 9. The target compound N-((1S)-2-((4-((1R,2S)-1-(5-((3,3-difluoroazacyclobutane-1-yl)methyl)oxazol-2-yl)-1-propamidopropane-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 14) was obtained.

[0237] LCMS(ESI)calcd for C 35 H 46 F3N7O4[M+H] + m / z 687.36, found 687.15.

[0238] 1H NMR (400MHz, DMSO) δ9.80(s,1H),8.45(dd,J=22.5,8.6Hz,2H),7.65(t,J=8.3Hz,1H),7.48(d,J=1.8Hz,1H),7.04(d ,J=12.0Hz,1H),6.95–6.88(m,2H),6.86(s,1H),5.42–5.31(m,1H),5.13(t,J=9.1Hz,1H),4.49(t,J=8.3Hz,1H),3.6 3(d,J=15.0Hz,2H),3.52(t,J=12.1Hz,4H),3.29–3.23(m,1H),2.20–2.11(m,2H),1.79(t,J=11.2Hz,2H),1.66-1.6 1(m,3H),1.37–1.33(m,6H),1.24(d,J=7.0Hz,4H),1.06(d,J=11.6Hz,1H),0.99–0.95(m,3H),0.85(d,J=6.4Hz,5H).

[0239] Example 13: N-((1S)-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl))amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 15)

[0240]

[0241] Steps 1 to 4 refer to the synthesis scheme of steps 3 to 6 in Example 1.

[0242] Steps 5 to 7 follow the synthetic scheme of steps 1 to 3 in Example 8 to obtain N-((1S)-2-((4-((1R,2S)-1-(5-(((dimethylamino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl))amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 15).

[0243] LCMS(ESI)calcd for C 34 H 48 FN7O4[M+H] + m / z 638.4, found 638.5.

[0244] 1H NMR (400MHz, DMSO) δ9.78(s,1H),8.50(d,J=8.8Hz,1H),8.42(d,J=8.4Hz,1H),7.67(t,J=8.4Hz,1H),7.48( d,J=2.0Hz,1H),7.02(d,J=12.4Hz,1H),6.95–6.86(m,2H),6.80(s,1H),5.42–5.33(m,1H),5.09(t,J=9.2Hz ,1H),4.49(t,J=8.4Hz,1H),3.38–3.32(m,2H),3.25–3.20(m,1H),2.22–2.10(m,2H),1.98(s,6H),1.82–1.5 8(m,5H),1.39–1.31(m,6H),1.28–1.10(m,5H),1.09–1.01(m,1H),0.98(t,J=7.6Hz,3H),0.92–0.81(m,5H).

[0245] Example 14: N-((1S)-2-((4-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 19)

[0246]

[0247] Steps 1 to 3 follow the synthetic scheme of steps 1 to 3 in Example 8, yielding the target compound N-((1S)-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 19). LC-MS (ESI) calcd for C 36 H 50 FN7O4[M+Na] + m / z 686.4, found 686.5. 1 H NMR (400MHz, DMSO-d6) δ9.77(s,1H),8.49(d,J=8.8Hz,1H),8.42(d,J=8.3Hz,1H),7.69(t,J=8.3Hz,1H),7.48(d,J=1.9Hz,1H),7.03(d ,J=12.1Hz,1H),6.93-6.87(m,2H),6.81(s,1H),5.43-5.33(m,1H),5.10(t,J=9.3Hz,1H),4.50(t,J=8.3Hz,1H),3.52(s,2H),3.27-3. 19(m,1H),2.22-2.09(m,2H),2.02(s,3H),1.86-1.75(m,2H),1.69(d,J=12.2Hz,2H),1.62(d,J=10.9Hz,1H),1.43-1.39(m,1H),1.37- 1.32(m,6H),1.30-1.19(m,5H),1.06(d,J=12.5Hz,1H),0.98(t,J=7.6Hz,3H),0.91-0.82(m,5H),0.35-0.29(m,2H),0.19-0.14(m,2H).

[0248] Example 15: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3,3,4,4-tetrafluoropyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 21)

[0249]

[0250] Step 1: N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (21a)

[0251] N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (1b) (1.20 g, 3.44 mmol) was dissolved in anhydrous methanol (10 mL) at room temperature, and sodium borohydride (0.26 g, 6.87 mmol) was added in portions with stirring. The reaction mixture was then allowed to react at room temperature for 1 hour before termination. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 80 / 1) to give the target compound N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (21a). LC-MS (ESI) calcd for C 16 H 18 FN3O5[M+H] + m / z 352.1,fo und 352.1.

[0252] Step 2: N-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (21b)

[0253] N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (21a) (1.10 g, 3.13 mmol) and imidazole (0.64 g, 9.39 mmol) were dissolved in N,N-dimethylformamide (10 mL) at room temperature, and tert-butyldiphenylchlorosilane (1.72 g, 6.26 mmol) was slowly added dropwise with stirring. The reaction mixture was then allowed to react at room temperature for 2 hours to terminate the reaction. The reaction mixture was diluted with water and extracted with dichloromethane. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to give the target compound N-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (21b). LC-MS (ESI) calculation for C 32 H 36 FN3O5Si[M+H] + m / z590.3, found 590.4.

[0254] Steps 3 to 6 follow the synthetic scheme of steps 5 to 8 in Example 9, yielding the target compound N-((S)-2-((4-(1R,2S)-1-(5-((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamido-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21f). LC-MS (ESI) calcd for C 48 H 61 FN6O5Si[M+H] + m / z 849.4, found 850.8.

[0255] Step 7: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamido-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21g)

[0256] N-((S)-2-((4-(1R,2S)-1-(5-((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamido-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxyethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21f) (0.08 g, 0.09 mmol) was dissolved in tetrahydrofuran (3 mL) at room temperature, and tetrabutylammonium fluoride (0.07 g, 0.27 mmol) was added with stirring. The reaction mixture was then allowed to react at room temperature for 1 hour before termination. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 50 / 1) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamido-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 g).

[0257] LC-MS(ESI)calcd for C 32 H 43 FN6O5[M+Na] + m / z 611.3, found 611.4.

[0258] Step 8: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21h)

[0259] N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamido-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 g) (0.05 g, 0.08 mmol) was dissolved in dichloromethane (3 mL) at room temperature, and Dess-Martin oxidant (0.17 g, 0.50 mmol) was added with stirring. The reaction mixture was then allowed to react at room temperature for 1 hour before termination. Saturated aqueous solutions of sodium thiosulfate and sodium bicarbonate were added to the reaction mixture, and the mixture was extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 60 / 1) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h). LC-MS (ESI) calcd for C 32 H 41 FN6O5[M+H] + m / z 609.3, found 609.4.

[0260] Step 9: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3,3,4,4-tetrafluoropyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 21)

[0261] 3,3,4,4-Tetrafluoropyrrolidine hydrochloride (0.010 g, 0.06 mmol) was dissolved in anhydrous methanol (6 mL) at room temperature, and triethylamine (0.020 g, 0.12 mmol) was added. The reaction mixture was then stirred before the addition of glacial acetic acid (0.020 g, 0.24 mmol) and N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h) (0.020 g, 0.03 mmol). The reaction mixture was heated to 30 °C and reacted for 1 h, followed by the addition of sodium cyanoborohydride (0.004 g, 0.06 mmol). The reaction was then terminated after 1 hour at room temperature. A saturated aqueous sodium bicarbonate solution was added to the reaction solution, followed by extraction with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by preparative high-performance liquid chromatography (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-(((3,3,4,4-tetrafluoropyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 21). LC-MS (ESI) calcd for C 36 H 46 F5N7O4[M+H] + m / z 736.4, found 736.5. 1 HNMR(400MHz, DMSO-d6)δ9.77(s,1H),8.49(d,J=8.8Hz,1H),8.40(d,J=8.2Hz,1H),7.66(t,J=8.3Hz,1H), 7.48(d,J=1.9Hz,1H),7.06(d,J=12.3Hz,1H),6.92-6.87(m,3H),5.43-5.31(m,1H),5.14(t,J=9.1Hz,1H), 4.49(t,J=8.1Hz,1H),3.76-3.68(m,2H),3.31-3.22(m,2H),3.12-3.02(m,4H),2.26-2.07(m,2H),1.83-1 .60(m,5H),1.37-1.32(m,6H),1.26-1.18(m,4H),1.11-1.03(m,1H),1.00-0.95(m,3H),0.91-0.82(m,5H).

[0262] Example 16: N-((1S)-2-((4-((1R,2S)-1-(5-((3-azabicyclo[3,1,0]hexane-3-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 22)

[0263]

[0264] Step 1: N-((1S)-2-((4-((1R,2S)-1-(5-((3-azabicyclo[3,1,0]hexane-3-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 22)

[0265] 3-azabicyclo[3,1,0]hexane hydrochloride (0.008 g, 0.065 mmol) was dissolved in methanol (3 mL) at room temperature. N,N-diisopropylethylamine (0.017 g, 0.13 mmol) was added with stirring and stirred at room temperature for 5 minutes. Acetic acid (0.015 g, 0.26 mmol) and N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h) (0.020 g, 0.032 mmol) were added sequentially. After reacting the reaction solution at 30°C for 2 hours, sodium cyanoborohydride (0.008 g, 0.13 mmol) was added. The reaction solution was then terminated by reacting at room temperature for 1 hour. The reaction solution was diluted with saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was purified by high performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((1S)-2-((4-((1R,2S)-1-(5-((3-azabicyclo[3,1,0]hexane-3-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 22).

[0266] LCMS(ESI)calcd for C 37 H50 FN7O4[M+H] + m / z 676.4, found 676.7.

[0267] 1 H NMR (400MHz, DMSO-d6) δ9.78(s,1H),8.49(d,J=8.9Hz,1H),8.41(d,J=8.4Hz,1H),7.70(t,J=8.2Hz,1H),7.48(d,J=1.8Hz,1H),7.03(d ,J=11.9Hz,1H),6.91(d,J=1.8Hz,1H),6.86(d,J=7.3Hz,1H),6.75(s,1H),5.43-5.32(m,1H),5.09(t,J=9.3Hz,1H),4.50(t,J=8.3Hz,1 H),3.54-3.44(m,2H),3.25-3.17(m,1H),2.62(d,J=8.4Hz,2H),2.21-2.08(m,4H),1.86-1.73(m,2H),1.71-1.55(m,3H),1.35(dd,J=9. 4, 6.7Hz, 6H), 1.24 (d, J = 6.9Hz, 7H), 1.13-1.02 (m, 1H), 0.98 (t, J = 7.6Hz, 3H), 0.90-0.80 (m, 5H), 0.55-0.47 (m, 1H), 0.25-0.17 (m, 1H).

[0268] Example 17: N-((1S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)pyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 23)

[0269]

[0270] Step 1: N-((1S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)pyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 23)

[0271] 3-(trifluoromethyl)pyrrolidine hydrochloride (0.01 g, 0.06 mmol) was dissolved in anhydrous methanol (2 mL) at room temperature. Then, N,N-diisopropylethylamine (0.02 g, 0.12 mmol) was added, followed by the addition of glacial acetic acid (0.02 g, 0.24 mmol) and N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h) (0.02 g, 0.03 mmol). The reaction mixture was then heated to 30 °C and reacted for 1 h. Sodium cyanoborohydride (0.004 g, 0.06 mmol) was then added. The reaction was then terminated after 1 hour at room temperature. The reaction solution was diluted with saturated sodium bicarbonate aqueous solution and extracted with ethyl acetate (3 × 20 mL). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was separated by high performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile). The product fraction was freeze-dried to obtain the target compound N-((1S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)pyrrolidone-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 23).

[0272] LC-MS(ESI)calcd for C 37 H 49 F4N7O4[M+H] + m / z 732.4, found 732.5.

[0273] 1H NMR (400MHz, DMSO-d6) δ9.79 (s, 1H), 8.52-8.46 (m, 1H), 8.41 (d, J = 8.1Hz, 1H), 7.65 (t, J = 8.3Hz, 1H), 7.48 (d, J = 1.9Hz, 1H), 7.08-7.00 (m, 1H),6.94-6.84(m,2H),6.82(s,1H),5.43-5.32(m,1H),5.12(t,J=9.0Hz,1H),4.49(t,J=8.2Hz,1H),3.59-3.48(m,2H),3.28-3.19(m,2H) ,3.03-2.95(m,1H),2.64-2.59(m,1H),2.47-2.43(m,1H),2.37-2.29 (m,2H),2.20-2.09(m,2H),1.97-1.89(m,1H),1.84-1.75(m,2H),1.7 3-1.65(m,3H),1.64-1.58(m,1H),1.37-1.32(m,6H),1.26-1.20(m,4H),1.06(d,J=12.0Hz,1H),0.98(t,J=7.6Hz,3H),0.91-0.82(m,5H).

[0274] Example 18: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((3-fluorozacyclobutan-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 24)

[0275]

[0276] Step 1: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((3-fluorozacricyclobutan-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 24)

[0277] 3-Fluorozylidene hydrochloride (0.010 g, 0.060 mmol) was dissolved in methanol (4 mL) at room temperature. N,N-diisopropylethylamine (0.031 g, 0.24 mmol) was added with stirring, and the mixture was stirred at room temperature for 5 minutes. Acetic acid (0.0288 g, 0.48 mmol) was then added. The reaction mixture was stirred at 30 °C for 1 hour, and then N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h) (0.020 g, 0.032 mmol) and sodium cyanoborohydride (0.008 g, 0.12 mmol) were added. The reaction mixture was then terminated after reacting at room temperature for 1 hour. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((3-fluoroazacyclobutan-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 24).

[0278] LC-MS(ESI)calcd for C 35 H 47 F2N7O4[M+H] + m / z 668.4, found 668.6.

[0279] 1H NMR (400MHz, DMSO-d6) δ9.80(s,1H),8.47(d,J=8.8Hz,1H),8.41(d,J=8.3Hz,1H),7.65(t,J=8.0Hz,1H),7.48(d,J=1.9 Hz,1H),7.03(d,J=12.2Hz,1H),6.93-6.90(m,2H),6.81(s,1H),5.40-5.36(m,1H),5.14-4.95(m,2H),4.49(t,J=5.2Hz ,1H),3.52(s,2H),3.47-3.21(m,4H),3.15-3.01(m,2H),2.18-2.11(m,2H),1.89-1.54(m,6H),1.36(d,J=6.8Hz,3H),1 .34(d,J=6.4Hz,3H),1.27-1.21(m,3H),1.20-1.18(s,1H),1.08-1.04(m,1H),0.97(t,J=7.6Hz,3H),0.88-0.83(m,4H).

[0280] Example 19: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)azacyclobutane-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 25)

[0281]

[0282] Step 1: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)azacyclobutane-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 25)

[0283] 3-(trifluoromethyl)azacyclobutane hydrochloride (0.015 g, 0.060 mmol) was dissolved in methanol (4 mL) at room temperature. N,N-diisopropylethylamine (0.031 g, 0.24 mmol) was added with stirring and stirred at room temperature for 5 minutes. Acetic acid (0.0288 g, 0.48 mmol) was then added. After stirring the reaction mixture at 30°C for 1 hour, N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (21 h) (0.020 g, 0.032 mmol) and sodium cyanoborohydride (0.008 g, 0.12 mmol) were added. The reaction mixture was then terminated after reacting at room temperature for 1 hour. The reaction mixture was absorbed with water and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude residue was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-((3-(trifluoromethyl)azacyclobutane-1-yl)methyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 25).

[0284] LC-MS(ESI)calcd for C 36 H 47 F4N7O4[M+H] + m / z 718.4, found 718.6.

[0285] 1H NMR (400MHz, DMSO-d6) δ9.79(s,1H),8.46(d,J=9.0Hz,1H),8.40(d,J=8.3Hz,1H),7.64(t,J=8.3Hz,1H),7.48 (d,J=1.9Hz,1H),7.03(d,J=12.0Hz,1H),6.93-6.86(m,2H),6.81(s,1H),5.40-5.36(m,1H),5.12(t,J=9.1Hz ,1H),4.49(t,J=8.4Hz,1H),3.49(s,2H),3.40-3.21(m,5H),3.13(s,2H),2.20-2.09(m,2H),1.85-1.58(m,5H ),1.35(dd,J=8.9,6.6Hz,6H),1.25-1.20(m,4H),1.08-1.04(m,1H),0.97(t,J=7.6Hz,3H),0.91-0.83(m,5H).

[0286] Example 20: N-((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 34)

[0287]

[0288] Steps 1 to 3 were performed according to the synthetic scheme of steps 7 to 9 in Example 2, yielding the target compound N-((S)-1-cycloheptyl-2-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 34).

[0289] LC-MS(ESI)calcd for C 36 H 50 FN7O4[M+H] + m / z 664.4, found 664.5.

[0290] 1H NMR (400MHz, DMSO-d6) δ9.78(s,1H),8.49(d,J=8.8Hz,1H),8.41(d,J=8.5Hz,1H),7.67(t,J=8.2Hz,1H),7.48(d,J=1. 8Hz,1H),7.03(d,J=12.0Hz,1H),6.93-6.87(m,2H),6.81(s,1H),5.42-5.32(m,1H),5.11(t,J=9.3Hz,1H),4.59(t,J=8 .3Hz,1H),3.52(s,2H),3.28-3.19(m,1H),2.23-2.07(m,3H),2.02(s,3H),1.71-1.60(m,4H),1.57-1.46(m,4H),1.44- 1.39(m,3H),1.38-1.32(m,8H),1.25(d,J=6.9Hz,3H),0.98(t,J=7.6Hz,3H),0.32(d,J=6.4Hz,2H),0.20-0.14(m,2H).

[0291] Example 21: N-((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (Compound 39)

[0292]

[0293] Step 1: ((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)tert-butyl carbamate (39a)

[0294] N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)propyl)propionamide (DM87-4) (0.08 g, 0.19 mmol) and Boc-L-cyclohexylglycine (0.10 g, 0.39 mmol) were dissolved in N,N-dimethylformamide (3 mL) with stirring. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.15 g, 0.39 mmol) and N,N-diisopropylethylamine (0.15 g, 1.17 mmol) were added sequentially. The reaction was terminated after 16 hours at 30 °C. The reaction solution was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 72 / 28) to obtain the target compound ((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)carbamate tert-butyl carbamate (39a).

[0295] LCMS(ESI)calcd for C 33 H 46 F3N5O5[M+H] + m / z 650.4, found 650.4.

[0296] Step 2: N-((1R,2S)-2-(4-((S)-2-amino-2-cyclohexylacetamyl)-3-fluorophenyl)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)propyl)propionamide (39b)

[0297] ((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)tert-butyl carbamate (39a) (0.060 g, 0.092 mmol) was dissolved in hydrogen chloride / 1,4-dioxane solution (4 M, 3 mL) under nitrogen protection and the reaction was terminated after 2 hours at room temperature. The reaction solution was concentrated to give the crude target compound N-((1R,2S)-2-(4-(((S)-2-amino-2-cyclohexylacetamido)-3-fluorophenyl)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)propyl)propionamide (39b). The product was used directly in the next step without further purification.

[0298] LCMS(ESI)calcd for C 28 H 38 F3N5O3[M+H] + m / z 550.3, found 550.4.

[0299] Step 3: N-((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (Compound 39)

[0300] At room temperature, N-((1R,2S)-2-(4-((S)-2-amino-2-cyclohexylacetamyl)-3-fluorophenyl)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)propyl)propionamide (39b) (0.050 g, crude) and 1-ethyl-1H-pyrazole-5-carboxylic acid (0.024 g, 0.17 mmol) were dissolved in N,N-dimethylformamide (3 mL). 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.065 g, 0.17 mmol) and N,N-diisopropylethylamine (0.066 g, 0.51 mmol) were added sequentially with stirring. The reaction was terminated after 16 hours at 30 °C. The reaction solution was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-1-cyclohexyl-2-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (compound 39).

[0301] LCMS(ESI)calcd for C 34 H 44 F3N7O4[M+H] + m / z 672.4, found 672.4.

[0302] 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),8.50(d,J=8.8Hz,1H),8.43(d,J=8.2Hz,1H),7.64(t,J=8.3Hz,1H),7.47(d,J=1.9Hz ,1H),7.05(d,J=12.0Hz,1H),6.99(d,J=2.0Hz,1H),6.89(d,J=7.4Hz,1H),6.84(s,1H),5.12(t,J=9.2Hz,1H),4.56-4.41( m,3H),3.65-3.52(m,2H),3.28-3.20(m,1H),2.79(t,J=13.0Hz,2H),2.49-2.40(m,2H),2.24-2.07(m,4H),1.89-1.76(m, 2H),1.75-1.66(m,2H),1.65-1.55(m,2H),1.32-1.23(m,6H),1.22-1.09(m,4H),1.09-1.01(m,1H),0.98(t,J=7.6Hz,3H).

[0303] Example 22: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-(2-methoxyacetamido)propyl-2-yl)-2-fluorophenyl)amino)-3-oxopropane-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 40)

[0304]

[0305] Step 1: N-((1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)-2-methoxyacetamide (40a)

[0306] 2-Methoxyacetic acid (0.07 g, 0.80 mmol) and (1R,2S)-1-(5-((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)prop-1-amine (7b) (0.17 g, 0.40 mmol) were dissolved in N,N-dimethylformamide (3 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.31 g, 0.80 mmol) and N,N-diisopropylethylamine (0.31 g, 2.42 mmol) were added sequentially. The reaction was terminated after 16 hours at 30 °C. The reaction solution was purified by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 55 / 45) to obtain the target compound N-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)-2-methoxyacetamide (40a).

[0307] LC-MS(ESI)calcd for C 20 H 23 F3N4O5[M+H] + m / z 457.2, found 457.2.

[0308] Steps 2 to 5 follow the synthetic scheme of steps 4 to 7 in Example 4 to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-(2-methoxyacetamido)propyl-2-yl)-2-fluorophenyl)amino)-3-oxopropane-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 40).

[0309] LC-MS(ESI)calcd for C 36 H 46 F3N7O5[M+Na] + m / z 736.3, found 736.6.

[0310] 1H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.39(dd,J=8.8,3.5Hz,2H),7.71(t,J=8.3Hz,1H),7.49(d,J=1.8Hz,1H),7.05(d ,J=12.2Hz,1H),6.94-6.87(m,2H),6.84(s,1H),5.46-5.31(m,1H),5.16(t,J=9.2Hz,1H),4.92(t,J=8.2Hz,1H),3.91 -3.83(m,2H),3.63-3.54(m,2H),3.43-3.35(m,2H),3.29(s,3H),2.81(t,J=13.2Hz,2H),2.22-2.11(m,2H),1.36(dd, J=14.8,6.6Hz,6H),1.26(d,J=6.9Hz,3H),0.94-0.80(m,2H),0.77-0.67(m,1H),0.49-0.41(m,1H),0.41-0.10(m,8H).

[0311] Example 23: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-ethyl-1H-pyrazole-5-carboxamide (Compound 42)

[0312]

[0313] Step 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-ethyl-1H-pyrazole-5-carboxamide (Compound 42)

[0314] (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (7f) (0.07 g, crude) and 1-ethyl-1H-pyrazole-5-carboxylic acid (0.033 g, 0.23 mmol) were dissolved in N,N-dimethylformamide (3 mL) with stirring. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.089 g, 0.23 mmol) and N,N-diisopropylethylamine (0.090 g, 0.70 mmol) were added after stirring at 30 °C for 16 hours. The reaction mixture was then diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-ethyl-1H-pyrazole-5-carboxamide (42).

[0315] LCMS(ESI)calcd for C 35 H 44 F3N7O4[M+H] + m / z 684.4, found 684.3.

[0316] 1H NMR (400MHz, DMSO-d6) δ9.85(s,1H),8.49(d,J=8.8Hz,1H),8.39(d,J=8.9Hz,1H),7.71(t,J=8.3Hz,1H),7.47(d,J=2.0Hz,1H),7.06( dd,J=12.2,1.6Hz,1H),6.97(d,J=2.0Hz,1H),6.90(d,J=8.4Hz,1H),6.83(s,1H),5.12(t,J=9.2Hz,1H),4.93(t,J=8.2Hz,1H),4.54-4 .41(m,2H),3.66-3.53(m,2H),3.29-3.19(m,1H),2.81(t,J=12.9Hz,2H),2.55-2.51(m,1H),2.49-2.43(m,1H),2.24-2.08(m,4H),1. 33-1.23(m,6H),0.97(t,J=7.6Hz,3H),0.93-0.78(m,2H),0.78-0.68(m,1H),0.49-0.41(m,1H),0.40-0.26(m,3H),0.25-0.10(m,4H).

[0317] Example 24: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 43)

[0318]

[0319] Step 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 43)

[0320] (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (7f) (0.07 g, crude) and 1-isopropyl-1H-pyrazole-5-carboxylic acid (0.036 g, 0.23 mmol) were dissolved in N,N-dimethylformamide (3 mL) with stirring. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.089 g, 0.23 mmol) and N,N-diisopropylethylamine (0.090 g, 0.70 mmol) were added after stirring. The reaction was terminated after 16 hours at 30 °C. The reaction solution was diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 43).

[0321] LCMS(ESI)calcd for C 36 H 46 F3N7O4[M+H] + m / z 698.4, found 698.4.

[0322] 1H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.49(d,J=8.9Hz,1H),8.38(d,J=8.8Hz,1H),7.72(t,J=8.3Hz,1H),7.49(d,J=1.8Hz,1H),7.06(d,J= 12.3Hz,1H),6.94-6.87(m,2H),6.84(s,1H),5.49-5.32(m,1H),5.12(t,J=9.2Hz,1H),4.92(t,J=8.2Hz,1H),3.64-3.51(m,2H),3.29-3.20 (m,1H),2.81(t,J=13.3Hz,2H),2.55-2.51(m,1H),2.48-2.43(m,1H),2.25-2.07(m,4H),1.36(dd,J=14.9,6.6Hz,6H),1.25(d,J=6.9Hz,3H ),0.97(t,J=7.6Hz,3H),0.94-0.86(m,1H),0.86-0.79(m,1H),0.77-0.68(m,1H),0.51-0.41(m,1H),0.40-0.26(m,3H),0.25-0.09(m,4H).

[0323] Example 25: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (Compound 45)

[0324]

[0325] Step 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (Compound 45)

[0326] (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (7f) (0.05 g, crude) and 4-methyl-1,2,5-oxadiazole-3-carboxylic acid (0.021 g, 0.17 mmol) were dissolved in N,N-dimethylformamide (3 mL) with stirring. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.064 g, 0.17 mmol) and N,N-diisopropylethylamine (0.065 g, 0.50 mmol) were added after stirring at 30 °C for 16 hours. The reaction mixture was then diluted with water and extracted with ethyl acetate. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated, and the crude product was purified by high-performance liquid chromatography (alkaline conditions, mobile phase containing ammonium bicarbonate) to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((3,3-difluoropyrrolidone-1-yl)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (compound 45).

[0327] LCMS(ESI)calcd for C 33 H 40 F3N7O5[M+H] + m / z 672.3, found 672.4.

[0328] 1H NMR (400MHz, DMSO-d6) δ9.91(s,1H),9.01(d,J=9.0Hz,1H),8.50(d,J=8.8Hz,1H),7.69(t,J=8.3Hz,1H),7.06(d, J=12.0Hz,1H),6.91(d,J=8.2Hz,1H),6.84(s,1H),5.12(t,J=9.1Hz,1H),5.05-4.95(m,1H),3.65-3.52(m,2H),3 .30(s,1H),3.29-3.20(m,1H),2.80(t,J=13.3Hz,2H),2.55-2.52(m,1H),2.48(s,3H),2.30-2.02(m,4H),1.25(d ,J=6.9Hz,3H),0.97(t,J=7.6Hz,3H),0.92-0.70(m,3H),0.51-0.43(m,1H),0.42-0.35(m,1H),0.34-0.16(m,6H).

[0329] Example 26: 1-Cyclopropyl-N-((S)-1,1-Dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1H-pyrazole-5-carboxamide (Compound 46)

[0330]

[0331] Step 1: 1-Cyclopropyl-N-((S)-1,1-Dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1H-pyrazole-5-carboxamide (Compound 46)

[0332] At room temperature, 1-cyclopropyl-1H-pyrazole-5-carboxylic acid (0.03 g, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.06 g, 0.15 mmol), and N,N-diisopropylethylamine (0.06 g, 0.46 mmol) were added to a stirred solution of (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f) (0.04 g, crude) in N,N-dimethylformamide (1 mL). The reaction was terminated after 16 hours at 30 °C. The reaction solution was filtered, and the filtrate was separated by high performance liquid preparative chromatography (mobile phase: 10 mM ammonium bicarbonate aqueous solution / acetonitrile = 1 / 1) to obtain 1-cyclopropyl-N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1H-pyrazole-5-carboxamide (compound 46).

[0333] LC-MS(ESI)calcd for C 36 H 46 FN7O4[M+H] + m / z 660.4, found 660.4

[0334] 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.48(d,J=8.8Hz,1H),8.41(d,J=8.9Hz,1H),7.72(t,J=8.3Hz,1H),7.41(d,J=1.8H z,1H),7.04(d,J=12.2Hz,1H),6.94-6.88(m,2H),6.81(s,1H),5.12(t,J=9.2Hz,1H),4.96(t,J=8.2Hz,1H),4.47-4.40(m ,1H),3.53(s,2H),3.28-3.20(m,2H),2.22-2.08(m,2H),2.05(s,3H),1.48-1.41(m,1H),1.25(d,J=6.9Hz,3H),1.14-1. 01(m,2H),0.97(t,J=7.6Hz,3H),0.94-0.78(m,4H),0.72(dd,J=17.5,9.0Hz,1H),0.49-0,42(m,1H),0.40-0.10(m,10H).

[0335] Example 27: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-methyl-1H-pyrazole-5-carboxamide (Compound 47)

[0336]

[0337] Step 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-methyl-1H-pyrazole-5-carboxamide (Compound 47)

[0338] At room temperature, 1-methyl-1H-pyrazole-5-carboxylic acid (0.02 g, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.06 g, 0.15 mmol), and N,N-diisopropylethylamine (0.06 g, 0.46 mmol) were added to a stirred solution of (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f) (0.04 g, crude) in N,N-dimethylformamide (1 mL). The reaction was terminated after 16 hours at 30 °C. The reaction solution was filtered, and the filtrate was separated by high-performance liquid chromatography (HPLC) preparative chromatography (mobile phase: 10 mM ammonium bicarbonate aqueous solution / acetonitrile = 1 / 1) to obtain N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-methyl-1H-pyrazole-5-carboxamide (compound 47). LC-MS (ESI) calcd for C 34 H 44 FN7O4[M+H] + m / z 634.4, found 634.6. 1H NMR (400MHz, DMSO-d6) δ9.85(s,1H),8.48(d,J=8.9Hz,1H),8.39(d,J=8.9Hz,1H),7.72(t,J=8.4Hz,1H),7.46(d ,J=2.0Hz,1H),7.03(t,J=7.2Hz,2H),6.92(d,J=8.3Hz,1H),6.80(s,1H),5.12(t,J=9.3Hz,1H),4.95(t,J=8.4H z,1H),4.03(s,3H),3.53(s,2H),3.29-3.21(m,1H),2.22-2.02(m,5H),1.48-1.41(m,1H),1.25(d,J=7.0Hz,3H) ,0.97(t,J=7.6Hz,3H),0.93-0.77(m,2H),0.72(dd,J=17.5,9.1Hz,1H),0.48-0.40(m,1H),0.36-0.11(m,11H).

[0339] Example 28: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (Compound 48)

[0340]

[0341] Step 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (Compound 48)

[0342] At room temperature, 4-methyl-1,2,5-oxadiazole-3-carboxylic acid (0.02 g, 0.19 mmol), 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.06 g, 0.15 mmol), and N,N-diisopropylethylamine (0.06 g, 0.46 mmol) were added to a stirred solution of (S)-2-amino-3,3-dicyclopropyl-N-(4-((1R,2S)-1-(5-((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)propionamide (6f) (0.04 g, crude) in N,N-dimethylformamide (1 mL). The reaction was terminated after 16 hours at 30 °C. The reaction solution was filtered, and the filtrate was separated by high-performance liquid chromatography (HPLC) preparative chromatography (mobile phase: 10 mM ammonium bicarbonate aqueous solution / acetonitrile = 1 / 1) to obtain N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(((cyclopropyl(methyl)amino)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-4-methyl-1,2,5-oxadiazole-3-carboxamide (compound 48). LC-MS (ESI) calcd for C 33 H 42 FN7O5[M+H] + m / z 636.3,fo und 634.6. 1 H NMR (400MHz, DMSO-d6) δ9.89(s,1H),9.02(d,J=9.0Hz,1H),8.48(d,J=8.9Hz,1H),7.71(t,J=8.3Hz,1H),7. 05(d,J=12.4Hz,1H),6.93(d,J=9.8Hz,1H),6.81(s,1H),5.12(t,J=9.2Hz,1H),5.02-4.97(m,1H),3.54(s,2 H),3.28-3.20(m,1H),2.48(s,3H),2.22-2.03(m,5H),1.49-1.40(m,1H),1.25(d,J=6.9Hz,3H),0.97(t,J= 7.6Hz,3H),0.92-0.78(m,2H),0.77-0.69(m,1H),0.51-0.43(m,1H),0.42-0.35(m,1H),0.35–0.16(m,10H).

[0343] Referring to the synthesis schemes of Examples 1 to 28, the following compounds were synthesized.

[0344]

[0345]

[0346]

[0347]

[0348]

[0349] To illustrate the beneficial effects of the present invention, the following experimental examples are provided.

[0350] Test Example 1: IL-17A / IL-17RA Binding Test

[0351] The purpose of this study was to evaluate the ability of the compound to bind IL-17A in vitro.

[0352] IL-17 modulator 4 (CAS: 2446803-65-0) was selected as the positive control compound. A 1X reaction buffer was prepared, and a 10 mM stock solution of the compound was prepared using DMSO. This stock solution was then serially diluted 3-fold to obtain 10 doses. 100 nL of the stock solution was transferred to the detection plate using ECHO. 2X IL-17A (Sino Biological, Cat#12047-H07B) solution was prepared, with 5 μL added to the compound well and the largest control well. The plate was centrifuged at 1000 rpm for 30 seconds and then incubated at room temperature for 15 min.

[0353] Prepare 2X IL-17RA (Sino Biological, Cat#10895-H41H-B) (5 μL / well) using buffer, centrifuge at 1000 rpm for 30 seconds, then add 5 μL of IL-17RA to the detection plate, centrifuge again at 1000 rpm, and incubate the detection plate at room temperature for 60 min. Prepare mixed beads (receptor:donor = 1:1) in buffer, add 10 μL to the detection plate, centrifuge at 1000 rpm for 30 seconds, and incubate at room temperature for 1 hour.

[0354] Readings were taken using Envision HTRF mode.

[0355] Inhibition percentage = 100 - (sample signal - minimum control group average) / (maximum control group average - minimum control group average) * 100

[0356] The compounds of the present invention have IC50 values ​​of <1 μM, and further IC50 values ​​of less than 100 nM.

[0357] According to the test results, the compound of the present invention has a good inhibitory effect on the binding of IL17A / RA protein in vitro.

[0358] Experimental Example 2: IL17A / AHEK-blue cell assay

[0359] The purpose of this experiment was to evaluate the activity of the compound at the cellular level.

[0360] IL-17 modulator 4 (CAS: 2446803-65-0) was selected as the positive control compound. The test compound was started at 10 mM and serially diluted 3-fold with DMSO for 10 doses. The positive control compound (IL-17 modulator 4, CAS: 2446803-65-0) was started at 1 mM and serially diluted 3-fold with DMSO for 10 doses to obtain a 1000× positive control and test compound. A 1000× negative control (100% DMSO) was also prepared.

[0361] Cell culture procedures were performed according to the manufacturer's instructions. HEK-Blue IL-17 cells (InvivoGen.Cat#hkb-il17) in the exponential growth phase were used for assay. Cell growth medium was aspirated, and cells were washed twice with PBS. Cells were resuspended in assay medium and diluted to the required density (15,000 cells / mL). Cells with a viability greater than 90% were used for assay. 25 nL of the compound dilution was transferred to a 384-well plate. 25 μL of HEK-Blue IL-17 cells and rhIL-17 protein (R&D, Cat#7955-IL) were seeded into 384-well plates. After culturing the cells at 37°C and 5% CO2 for 20 hours, 2 μL of cell supernatant was transferred to a 384-well assay plate. 20 μL of pNPP substrate was added, and absorbance at 405 nm was measured using an Envision 2105 multilabel reader.

[0362] Calculate the RLU signal (LUMcmpd) for each aperture.

[0363] The formula for calculating the inhibition rate is as follows:

[0364] %inhibition = 100 - (Signal) cmpd -Signal Ave_PC ) / (Signal Ave_VC -Signal Ave_PC )×100.

[0365] Signal ave_pc The average signal value of the positive control.

[0366] Signal ave_vc The average signal value of the negative control.

[0367] Computing IC 50And plot the effect-dose curve:

[0368] Using Graphpad 8.0, the %inhibition value and the logarithm of the compound concentration were fitted to a nonlinear regression (dose response-slope) to calculate the IC. 50 .

[0369] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0370] X: Logarithm of the inhibitory concentration; Y: Inhibition rate.

[0371] The results of some of the embodiments are shown in Table 1.

[0372] Table 1 Cell Inhibition Activity

[0373]

[0374]

[0375] The above results indicate that the compound of the present invention has a strong inhibitory effect on IL17 cells in vitro.

[0376] In summary, the compounds of this invention exhibit good IL-17A inhibitory activity and can be used to prepare IL-17 inhibitors, providing a new pharmaceutical possibility for the clinical treatment of diseases associated with abnormal IL-17 activity (such as plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, erythrodermic psoriasis, and palmoplantar psoriasis).

Claims

1. A compound of Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: in: n is selected from 0 or 1; When n = 0, ring A is selected from an optional substituted 5-membered heteroaromatic ring, wherein ring A is independently substituted by one or more substituents selected from the following: C 1-6 Alkyl and C 3-5 cycloalkyl; wherein the C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, halogen, and C. 3-6 cycloalkyl; When n=1, ring A is selected from an optionally substituted benzene ring or a 6-membered heteroaromatic ring, wherein the 6-membered heteroaromatic ring is an optionally substituted 6-membered heteroaromatic ring containing 1 or 2 nitrogen atoms; wherein ring A is independently substituted by one or more substituents selected from the following: halogen, -OR 11 -CN, -NHS(O)2CH3 and C 1-6 Alkyl; wherein the C 1-6 The alkyl group may optionally be substituted with one or more of the following substituents: hydrogen and halogen; R 11 Independently selected from: hydrogen and C 1-6 Alkyl, wherein the C 1-6 The alkyl group is independently substituted by one or more substituents selected from the following: hydrogen and halogen; R' and R'' are independently selected from: F and methyl; or R' and R'' can be linked together to form a cyclopropyl group; R 1 and R 2 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen; or the R 1 and R 2 C formed by combining with the same carbon 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is substituted with one or more substituents selected from the following: hydrogen and halogen; R 3 Selected from: methyl, halomethyl, ethyl, haloethyl, and methoxymethyl; R 4 Selected from hydrogen; R 5 Selected from methyl, halomethyl, ethyl, methoxymethyl, haloethyl, and: X is selected from: C(R) 10 ), nitrogen, oxygen, and sulfur; Y and Z are each independently selected from: nitrogen, oxygen, and sulfur; wherein R 10 Selected from: hydrogen, fluorine and C 1-6 alkyl; m is selected from 0, 1, or 2; R 6 R 7 Each element is independently selected from hydrogen, halogens, and carbon. 1-6 Alkyl and C 1-6 Halogenated alkyl; or the R 6 and R 7 They can connect and combine to form C 3-7 cycloalkyl; R 8 and R 9 Each is independently selected from hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen, halogen, C 1-6 Alkyl and C 3-7 cycloalkyl; Or R 8 and R 9 They can be linked together to form 3 to 7-membered monoheterocyclic alkyl groups, 5 to 12-membered bridged heterocyclic alkyl groups, or 5 to 12-membered spirocyclic heterocyclic alkyl groups. The 3- to 7-membered monoheterocyclic alkyl, 5- to 12-membered bridged-ring heterocyclic alkyl, or 5- to 12-membered spirocyclic heterocyclic alkyl are mentioned. Each may be optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen and halogen.

2. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, When n = 0, ring A is selected from: When n=1, ring A is selected from:

3. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 1 and R 2 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein the C 1-6 Alkyl and C 3-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen; in Selected from: or the R 1 and R 2 Combines with the same carbon to form C 5-7 Cycloalkyl group, the C 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is substituted with one or more substituents selected from: hydrogen and halogen; which are selected from:

4. The compound according to claim 3, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 1 and R 2 Combines with the same carbon to form C 5-7 Cycloalkyl group, the C 5-7 The cycloalkyl group is optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-3 Alkyl groups; selected from:

5. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 3 Selected from methyl; R 4 Selected from hydrogen.

6. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 5 Selected from ethyl, methoxymethyl and:

7. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, Selected from:

8. The compound according to claim 7, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, m is selected from 1; R 6 and R 7 Each is independently selected from hydrogen, fluorine, methyl, and trifluoromethyl; or the R 6 and R 7 They can combine to form cyclopropane.

9. The compound according to claim 8, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 8 and R 9 Each is independently selected from hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; Wherein C 1-6 Alkyl and C 3-7 The cycloalkyl group may optionally be substituted with one or more substituents selected from the following: hydrogen, halogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein Selected from: Or R 8 and R 9 They can be linked together to form 3 to 7-membered monoheterocyclic alkyl groups, 5 to 12-membered bridged heterocyclic alkyl groups, or 5 to 12-membered spirocyclic heterocyclic alkyl groups. The 3- to 7-membered monoheterocyclic alkyl, 5- to 12-membered bridged-ring heterocyclic alkyl, or 5- to 12-membered spirocyclic heterocyclic alkyl are mentioned. Each may be optionally substituted with one or more substituents selected from the following: hydrogen, halogen, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is optionally substituted with one or more substituents selected from the following: hydrogen and halogen; The above Selected from:

10. The compound according to claim 9, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The Selected from:

11. The compound, or its stereoisomer, or its pharmaceutically acceptable salt according to any one of claims 1-10, characterized in that, The compound described in Formula I is represented by Formula II: Among them, rings A and R 1 R 2 R 5 R 6 R 7 R 8 R 9 R`, R``, m, n, X, Y and Z as described in claim 1.

12. The compound according to claim 11, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by formula III: Among them B 1 B 2 B 3 B 4 and B 5 Each is independently selected from: CR 16 and nitrogen; Wherein R 16 Selected from: hydrogen, halogen, -OR 11 -CN, -NHS(O)2CH3 and C 1-6 alkyl; And B 1 B 2 B 3 B 4 and B 5 No more than two of them are selected from nitrogen, and the rest are CR. 16 ; R 1 R 2 R 5 R 6 R 7 R 8 R 9 R 11 X, Y and Z are as described in claim 1.

13. The compound according to claim 11, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by formula IV: Where C 1 C 2 C 3 and C 4 Each is independently selected from: CR 17 NR 18 N and oxygen; Wherein R 17 and R 18 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-5 cycloalkyl; Wherein C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, halogen, and C. 3-6 cycloalkyl; And C 1 C 2 C 3 and C 4 At least one of them is selected from NR 18 , N or oxygen, the rest are CR 17 ; R 1 R 2 R 5 R 6 R 7 R 8 R 9 X, Y and Z are as described in claim 1.

14. The compound according to claim 13, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by formula V: Where R 18 Selected from C 1-6 Alkyl and C 3-5 cycloalkyl, wherein the C 1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, C 1-6 Alkyl or C 3-6 cycloalkyl; R 1 R 2 R 5 R 6 R 7 R 8 R 9 X, Y and Z are as described in claim 1.

15. The compound according to any one of claims 1-14, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the compound is selected from any one of the following compounds:

16. A pharmaceutical composition, wherein, It includes the compound, stereoisomer or pharmaceutically acceptable salt thereof of any one of claims 1-15, and one or more pharmaceutically acceptable carriers, diluents or excipients.

17. Use of the compound of any one of claims 1-15 or a pharmaceutically acceptable salt thereof, or a solvate, metabolite, cocrystal or prodrug thereof, or the pharmaceutical composition of claim 16 in the preparation of a medicament for treating / preventing IL-17A-mediated diseases.

18. The use according to claim 17, wherein the disease includes plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, erythrodermic psoriasis, and palmoplantar psoriasis.

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