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 202411123341.8, 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 (European 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 SEFIR 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] In a first aspect, the present invention provides a compound of Formula I, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0007]

[0008] Wherein: ring A is selected from an optionally 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;

[0009] R 1 and R 2Each 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;

[0010] 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;

[0011] R 3 Selected from: methyl, halomethyl, ethyl, haloethyl, and methoxymethyl; R 4 Selected from hydrogen;

[0012] R 5 Selected from C 1-6 Alkyl; wherein the C 1-6 Alkyl groups are optionally substituted with one or more substituents selected from: halogens and -OR. 12 The R 12 Each is 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;

[0013] X is selected from: C(R) 8 ), nitrogen, oxygen, and sulfur; Y and Z are each independently selected from: nitrogen, oxygen, and sulfur; wherein R 8 Selected from: hydrogen, fluorine and C 1-6 alkyl;

[0014] R 6 and R 7 Each is independently selected from hydrogen, C 1-6 Alkyl, C 3-5 cycloalkyl, 5- to 6-membered heteroaryl and aryl; wherein the C 1-6 Alkyl, C 3-5 The cycloalkyl, 5- to 6-membered heteroaryl, and aryl groups are optionally substituted by one or more substituents selected from the following: hydrogen, halogen, C 1-3 Alkyl and C 3-5 cycloalkyl;

[0015] Or R 6 and R 7They 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, cyano, 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.

[0016] In some implementation schemes, ring A is selected from:

[0017]

[0018] In some implementation schemes, R 1 and R 2 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-7 cycloalkyl; wherein Selected from:

[0019] In some implementation schemes, 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; preferably derived from:

[0020]

[0021] In some implementation schemes, R 3 Selected from methyl; R 4 Selected from hydrogen.

[0022] In some implementation schemes, R 5 Preferably derived from ethyl, haloethyl, or methoxymethyl.

[0023] In some implementation schemes, Selected from:

[0024]

[0025] In some implementation schemes, R 6 and R 7 Each is independently selected from hydrogen, C 1-6 Alkyl, C 3-5 cycloalkyl, 5- to 6-membered heteroaryl and aryl; wherein the C 1-6 Alkyl, C 3-5The cycloalkyl, 5- to 6-membered heteroaryl, and aryl groups are optionally substituted by one or more substituents selected from the following: hydrogen, halogen, C 1-3 Alkyl and C 3-5 cycloalkyl; wherein the R 6 and R 7 Each of the following is independently selected from: hydrogen, methyl, deuterated methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, C 1-3 Alkylcyclopropyl, aryl, pyridine, oxazole, isoxazole and their corresponding halogenated compounds and C 1-3 Alkane-substituted compounds.

[0026] In some implementation schemes, R 6 and R 7 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.

[0027] 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, cyano, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is optionally substituted with one or more substituents selected from: hydrogen and halogen; wherein... Selected from:

[0028]

[0029] In some implementations, the Selected from:

[0030]

[0031] In some embodiments, the compound is represented by Formula II:

[0032]

[0033] Where C 1 C 2 C 3 and C 4 Each is independently selected from: CR 15 NR 16 N and oxygen;

[0034] Wherein R 15 and R 16 Each is independently selected from: hydrogen, C 1-6 Alkyl and C 3-5 cycloalkyl;

[0035] Wherein C1-6 Alkyl groups may optionally be substituted with one or more of the following substituents: hydrogen, halogen, and C. 3-6 cycloalkyl;

[0036] And C 1 C 2 C 3 and C 4 Two of them are selected from NR 16 One of them is selected from oxygen or CR 15 The rest are CR 15 ;

[0037] R 1 R 2 R 5 R 6 R 7 X, Y, and Z are as described in the first aspect of the present invention.

[0038] In some embodiments, the compound is represented by Formula III:

[0039]

[0040] Wherein R 16 It is methyl, ethyl, isopropyl, or fluoroethyl, R 1 R 2 It is a three-membered cycloalkyl group or R 1 R 2 It combines with the same carbon to form a six-membered cycloalkyl group, wherein the six-membered cycloalkyl group is optionally bonded to the same carbon atom. 1-3 Alkyl substitution, R 5 It is ethyl, haloethyl, or methoxymethyl, R 6 R 7 As described in the first aspect of the present invention.

[0041] In some embodiments, the compounds are selected from any of the following:

[0042]

[0043] 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.

[0044] 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 composition described in the second aspect, in the preparation of a medicament for treating / preventing IL-17A-mediated diseases.

[0045] The diseases mentioned are selected from plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, erythrodermic psoriasis, and palmoplantar psoriasis. Detailed Implementation

[0046] Detailed description of the invention

[0047] Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0048] "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.

[0049] “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.

[0050] “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.

[0051] “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).

[0052] "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).

[0053] "Halogen" indicates fluorine, chlorine, bromine or iodine, preferably fluorine or chlorine.

[0054] 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.

[0055] 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.

[0056] 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.

[0057] 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.

[0058] "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.

[0059] 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.

[0060] "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.

[0061] "Enantiomers" refer to two non-overlapping but mirror-image isomers of a compound.

[0062] 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.

[0063] 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.

[0064] 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).

[0065] 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 14 Compounds 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.

[0066] "Pharmaceutically acceptable salts" refer to those salts that retain the bioavailability and properties of the parent compound. These salts include:

[0067] (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.

[0068] (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.

[0069] "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.

[0070] "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.

[0071] "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.

[0072] 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.

[0073] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. 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 of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0074] The structure of the compounds was determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). NMR was 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-MS 2020 (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.

[0075] 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.

[0076] Example 1: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(3,3-difluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropane-2-yl)-2-fluorophenyl)amino)-3-oxopropane-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 1)

[0077]

[0078] Step 1: ((2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-oxo-1-(prop-2-yn-1-ylamino)but-2-yl)carbamate tert-butyl ester

[0079] At room temperature, (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (1a) (1.20 g, 3.50 mmol), 2-propyn-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-(prop-2-yn-1-ylamino)but-2-yl)carbamate tert-butyl ester (1b). LCMS(ESI)calcd forC 18 H 22 FN3O5[M+H] + m / z 380.2, found 380.1. 1 HNMR(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).

[0080] Step 2: Tert-butyl carbamate ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)carbamate was dissolved in DCM (50 mL) at room temperature under an oxygen atmosphere. The mixture was 1.20 g (3.20 mmol), PhI(OAc)2 (0.10 g, 0.03 mmol), and lithium iodide (0.04 g, 0.03 mmol). The reaction was terminated after 16 hours at room temperature under irradiation with a 40W household fluorescent bulb. After the reaction, the mixture was extracted with ethyl acetate, 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 ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)carbamate tert-butyl ester (1c). 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).

[0081] Step 3: 2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-carboxaldehyde (1d)

[0082] ((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)tert-butyl carbamate (1c) (1.90 g, 4.83 mmol) was dissolved in hydrogen chloride / 1,4-dioxane solution (4 M, 20 mL) at room temperature and stirred for 1 hour under nitrogen protection at room temperature until the reaction was terminated. The reaction solution was concentrated to give crude 2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-carboxaldehyde (1d), which was used directly in the next step without further purification. LC-MS (ESI) calcd for C 13 H 12 FN3O4[M+H] + m / z 294.1, found 294.1.

[0083] Step 4: N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)propionamide (1e)

[0084] 2-((1R,2S)-1-amino-2-(3-fluoro-4-nitrophenyl)propyl)oxazol-5-carboxaldehyde (1d) (1.90 g, crude) and N,N-diisopropylethylamine (4.47 g, 34.58 mmol) were dissolved together in dichloromethane (30 mL) at room temperature, and propionic anhydride (1.12 g, 8.64 mmol) was added with stirring. The reaction was terminated after 1 hour at room temperature. The reaction solution was diluted with water and extracted with dichloromethane. The solutions were combined, dried, and concentrated. The crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 60 / 1) to give the target compound N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-formyloxazol-2-yl)propyl)propionamide (1e). LC-MS (ESI) calcd for C 16 H 16 FN3O5[M+H] + m / z 350.1, found 350.0.

[0085] Step 5: N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (1f)

[0086] N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (1e) (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 stirred at room temperature for 1 hour to terminate the reaction. The reaction mixture was diluted with water and extracted with dichloromethane. The samples were combined, dried, and concentrated. 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 (1f). LC-MS (ESI) calcd for C 16 H 18 FN3O5[M+H] + m / z 352.1, found 352.1.

[0087] Step 6: N-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1g)

[0088] N-((1R,2S)-2-(3-fluoro-4-nitrophenyl)-1-(5-(hydroxymethyl)oxazol-2-yl)propyl)propionamide (1f) (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 stirred at room temperature for 2 hours to terminate the reaction. The reaction mixture was diluted with water and then extracted with dichloromethane. The organic phases were combined, dried, and concentrated. The crude product was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 5 / 1) to obtain the target compound N-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1 g). LC-MS (ESI) calculation for C 32 H 36 FN3O5Si[M+H] + m / z 590.3, found 590.4.

[0089] Step 7: N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)propyl)propionamide (1h)

[0090] N-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-2-(3-fluoro-4-nitrophenyl)propyl)propionamide (1 g) (1.40 g, 2.37 mmol) was dissolved in a tetrahydrofuran / water (3 / 1, 20 mL) mixture at room temperature. Zinc powder (1.55 g, 23.74 mmol) and ammonium chloride (2.54 g, 47.48 mmol) were added with stirring. The reaction was terminated after 2 hours at 80 °C under a nitrogen atmosphere. The reaction solution was cooled to room temperature, neutralized with saturated sodium bicarbonate solution, and then extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 50 / 1) to obtain the target compound N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)propyl)propionamide (1 h). LC-MS (ESI) calcd for C 32 H 38 FN3O3Si[M+H]+ m / z 560.3, found 560.4.

[0091] Step 8: ((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)tert-butyl carbamate (1i)

[0092] (S)-2-((tert-Butoxycarbonyl)amino)-3,3-dicyclopropylpropionic acid (1.15 g, 4.28 mmol) and N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-(((tert-Butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)propyl)propionamide (1 h) (1.20 g, 2.14 mmol) were dissolved in an 8 mL microwave-safe tube containing N,N-dimethylformamide. 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (1.63 g, 4.28 mmol) and N,N-diisopropylethylamine (1.66 g, 12.86 mmol) were added sequentially with stirring. The microwave-safe tube was sealed and the reaction was terminated at 30 °C for 16 hours. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was separated and concentrated by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 70 / 30), yielding the target compound ((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)carbamate (1i). LC-MS (ESI) calculation for C 46 H 59 FN4O6Si[M+H] + m / z 811.4,foun d 811.5.

[0093] Step 9: (S)-2-amino-N-(4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)-3,3-dicyclopropylpropionamide (1j)

[0094] ((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)carbamate (1i) (1.30 g, 1.60 mmol) was dissolved in dichloromethane (50 mL) at room temperature, and trifluoroacetic acid (6.22 g, 64.11 mmol) was slowly added dropwise under nitrogen protection. The reaction mixture was then stirred at room temperature for 2 hours to terminate the reaction. The reaction mixture was diluted with water, and sodium bicarbonate solid was added in portions until the pH was approximately 8. The resulting mixture was extracted with dichloromethane. The organic phases were combined, dried, and concentrated. The crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 50 / 1) to obtain the target compound (S)-2-amino-N-(4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)-3,3-dicyclopropylpropionamide (1j). LC-MS (ESI) calcd for C 41 H 51 FN4O4Si[M+H] + m / z711.4, found 711.8.

[0095] Step 10: N-((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1k)

[0096] At room temperature, 1-isopropyl-1H-pyrazole-5-carboxylic acid (0.06 g, 0.39 mmol) and (S)-2-amino-N-(4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)-3,3-dicyclopropylpropionamide (1j) (0.14 g, 0.20 mmol) were dissolved in a microwave-safe tube containing N,N-dimethylformamide (2 mL). 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.15 g, 0.39 mmol) and N,N-diisopropylethylamine (0.15 g, 1.18 mmol) were added sequentially with stirring. The microwave-safe tube was sealed, and the reaction was terminated after 16 hours at 30 °C. The reaction solution was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated to obtain a crude product. This product was then purified by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 63 / 37), concentrated, and the target compound N-((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1k). LC-MS (ESI) calculation for C 48 H 59 FN6O5Si[M+H] + m / z 847.4, found 847.8.

[0097] Step 11: N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1l)

[0098] N-((S)-1-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-3,3-dicyclopropyl-1-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1k) (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 stirred at room temperature for 1 hour to terminate the reaction. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 50 / 1) to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1l). LC-MS (ESI) calcd for C 32 H 41 FN6O5[M+Na] + m / z 631.3, found 631.4.

[0099] Step 12: N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1m)

[0100] N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-(hydroxymethyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1 L) (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 stirred at room temperature for 1 hour to terminate the reaction. The reaction was quenched by the slow addition of saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate solutions, followed by extraction with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was separated by silica gel column chromatography (eluent: dichloromethane / methanol = 60 / 1) to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1m). LC-MS (ESI) calcd for C 32 H 39 FN6O5[M+H] + m / z607.3, found 607.4.

[0101] Step 13: 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazole-5-carboxylic acid (1n)

[0102] 2-Methyl-2-butene (0.04 g, 0.53 mmol) and N-((S)-1,1-dicyclopropyl-3-((2-fluoro-4-((1R,2S)-1-(5-formyloxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (1 M) (0.04 g, 0.06 mmol) were dissolved in tert-butanol / tetrahydrofuran (1 / 1, 6 mL) at room temperature. A mixed aqueous solution (1 mL) of sodium chlorite (0.02 g, 0.20 mmol) and sodium dihydrogen phosphate (0.03 g, 0.20 mmol) was added to the reaction solution at room temperature. Stirring was continued for 10 minutes at room temperature until the reaction was stopped. The pH of the reaction solution was adjusted to 2 with dilute hydrochloric acid (1 M), and then extracted with ethyl acetate. The organic phases were combined, dried, and concentrated to obtain the crude target compound 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazole-5-carboxylic acid (1n). The product was used directly in the next step without further purification. LC-MS (ESI) calcd for C 32 H 39 FN6O6[M+H] + m / z 623.3, found 623.4.

[0103] Step 14: N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(3,3-difluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropane-2-yl)-2-fluorophenyl)amino)-3-oxopropane-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 1)

[0104] 3,3-Difluoropyrrolidine hydrochloride (0.07 g, 0.044 mmol) and 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (1n) (0.01 g, crude) were dissolved in N,N-dimethylformamide (3 mL) at room temperature. Then, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.02 g, 0.04 mmol) and N,N-diisopropylethylamine (0.02 g, 0.13 mmol) were added sequentially. The reaction mixture was then stirred at room temperature for 16 hours until the reaction was terminated. The reaction mixture was concentrated, diluted with water, extracted with dichloromethane, combined, dried, and the organic phase was concentrated. The crude product was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile). The obtained product fraction was freeze-dried to obtain the target compound N-((S)-1,1-dicyclopropyl-3-((4-((1R,2S)-1-(5-(3,3-difluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropane-2-yl)-2-fluorophenyl)amino)-3-oxopropane-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 1). LC-MS (ESI) calcd for C 36 H 44 F3N7O5[M+H] + m / z 712.3, found 712.5. 1HNMR(400MHz, DMSO-d6)δ9.86(s,1H),8.59(d,J=6.7Hz,1H),8.37(d,J=8.8Hz,1H),7.71(t,J=8.3Hz,1H),7.64(s,1H),7.49(d,J=1.9Hz,1H),7.14 (dd,J=12.2,1.7Hz,1H),6.94(d,J=7.7Hz,1H),6.90(d,J=1.9Hz,1H),5.4 4-5.34(m,1H),5.25-5.15(m,1H),4.91(t,J=8.2Hz,1H),4.15-3.95(m,1H ),3.87-3.70(m,2H),3.68-3.59(m,1H),3.38-3.34(m,1H),2.49-2.34(m, 2H),2.26-2.09(m,2H),1.43-1.33(m,6H),1.29(d,J=7.0Hz,3H),0.98(t, J=7.6Hz,3H),0.94-0.87(m,1H),0.86-0.78(m,1H),0.77-0.68(m,1H),0. 49-0.41(m,1H),0.40-0.26(m,3H),0.26-0.16(m,3H),0.15-0.09(m,1H).

[0105] Example 2: 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (Compound 2)

[0106]

[0107] Step 1: 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (Compound 2)

[0108] Methylamine hydrochloride (0.003 g, 0.048 mmol) and 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (1n) (0.015 g, crude) were dissolved in N,N-dimethylformamide (2 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.018 g, 0.048 mmol) and N,N-diisopropylethylamine (0.019 g, 0.144 mmol) were then added sequentially. The reaction mixture was then reacted at 30 °C for 16 hours, after which the reaction was terminated. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude residue was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (compound 2). LC-MS (ESI) calcd for C 33 H 42 FN7O5[M+H] + m / z 636.3, found 636.4. 1 H NMR (400MHz, DMSO-d6) δ9.84 (s, 1H), 8.52 (d, J = 8.8Hz, 1H), 8.38 (d, J = 8.9Hz, 1H), 8.35-8.30 (m, 1H), 7.73 (t, J = 8.4Hz, 1H), 7.5 2(s,1H),7.49(d,J=1.8Hz,1H),7.08(d,J=12.2Hz,1H),6.96(d,J=8.4Hz,1H),6.90(d,J=1.9Hz,1H),5.44-5.35(m,1H),5.20(t, J=8.8Hz,1H),4.91(t,J=8.2Hz,1H),2.71(d,J=4.6Hz,3H),2.26-2.09(m,2H),1.39-1.33(m,6H),1.26(d,J=7.0Hz,3H),0.97(t, J=7.6Hz,3H),0.93-0.86(m,1H),0.85-0.79(m,1H),0.76-0.68(m,1H),0.48-0.43(m,1H),0.42-0.25(m,4H),0.24-0.11(m,4H).

[0109] Example 3: N-cyclopropyl-2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazole-5-carboxamide (Compound 3)

[0110]

[0111] Step 1: N-cyclopropyl-2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxamide (Compound 3)

[0112] Cyclopropylamine (0.003 g, 0.048 mmol) and 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (1n) (0.015 g, crude) were dissolved in N,N-dimethylformamide (2 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.018 g, 0.048 mmol) and N,N-diisopropylethylamine (0.019 g, 0.144 mmol) were then added sequentially. The reaction mixture was then reacted at 30 °C for 16 hours, after which the reaction was terminated. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude residue was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound N-cyclopropyl-2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxamide (compound 3). LC-MS (ESI) calcd for C 35 H 44 FN7O5[M+H] + m / z 662.4, found 662.4. 1H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.52(d,J=8.8Hz,1H),8.39(dd,J=12.7,6.3Hz,2H),7.72(t,J=8.4Hz,1H),7.53(s,1H),7.49(d,J= 1.8Hz,1H),7.09(d,J=12.2Hz,1H),6.95(d,J=8.3Hz,1H),6.90(d,J=2.0Hz,1H),5.45-5.34(m,1H),5.20(t,J=8.8Hz,1H),4.91(t,J=8.2 Hz,1H),2.75-2.69(m,1H),2.24-2.07(m,2H),1.36(dd,J=13.0,6.6Hz,6H),1.25(d,J=7.0Hz,3H),0.97(t,J=7.6Hz,3H),0.93-0.87(m,1 H),0.86-0.78(m,1H),0.76-0.70(m,1H),0.70-0.64(m,2H),0.54-0.49(m,2H),0.48-0.42(m,1H),0.41-0.24(m,4H),0.23-0.09(m,4H).

[0113] Example 4: 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)-N-(pyridin-3-yl)oxazol-5-carboxamide (Compound 4)

[0114]

[0115] At room temperature, pyridine-3-amine (0.005 g, 0.048 mmol) and 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (1n) (0.015 g, crude) were dissolved in N,N-dimethylformamide (2 mL). 2-(7-azabenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate (0.018 g, 0.048 mmol) and N,N-diisopropylethylamine (0.019 g, 0.144 mmol) were added sequentially with stirring. The reaction mixture was then stirred at 30 °C for 16 hours until the reaction was terminated. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. 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 2-((1R,2S)-2-(4-((S)-3,3-dicyclopropyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)-1-propamidopropyl)-N-(pyridin-3-yl)oxazol-5-carboxamide (compound 4). LC-MS (ESI) calcd for C 37 H 43 FN8O5[M+H] + m / z 699.3, found 699.4.

[0116] 1 H NMR (400MHz, Methanol-d4) δ8.82(s,1H),8.29(s,1H),8.17(d,J=8.2Hz,1H),7.81(t,J=8.2Hz,1H),7. 72(s,1H),7.51(d,J=1.9Hz,1H),7.44-7.39(m,1H),7.08-7.00(m,2H),6.75(d,J=1.9Hz,1H),5.39-5. 32(m,2H),4.92-4.89(m,2H),3.49-3.39(m,3H),2.37-2.25(m,2H),1.45-1.40(m,9H),1.13(t,J=7.6H z,3H),0.86-0.75(m,2H),0.74-0.67(m,1H),0.49-0.38(m,3H),0.35-0.28(m,2H),0.27-0.10(m,4H).

[0117] Example 5: N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (Compound 5)

[0118]

[0119] Step 1: ((S)-2-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)tert-butyl carbamate (5a)

[0120] (S)-2-((tert-Butoxycarbonyl)amino)-2-((1R,4S)-4-methylcyclohexyl)acetic acid (0.87 g, 3.22 mmol) and N-((1R,2S)-2-(4-amino-3-fluorophenyl)-1-(5-(((tert-Butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)propyl)propionamide (1 h) (0.90 g, 1.61 mmol) were dissolved in N,N-dimethylformamide (4 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N′,N'-tetramethylurea hexafluorophosphate (1.22 g, 3.22 mmol) and N,N-diisopropylethylamine (1.25 g, 9.65 mmol) were added sequentially. The reaction was terminated after 16 hours at 30 °C. After cooling to room temperature, the reaction solution was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was separated by reversed-phase C18 column chromatography (mobile phase: acetonitrile / 0.1% formic acid aqueous solution = 9 1 / 9) to obtain the target compound ((S)-2-((4-((1R,2S)-1-(5-(((tert-butyldiphenylsilyl)oxy)methyl)oxazol-2-yl)-1-propamidopropyl-2-yl)-2-fluorophenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate tert-butyl ester (5a). LC-MS (ESI) calculation for C 46 H 61 FN4O6Si[M+H] + m / z 813.4, found 813.5.

[0121] Steps 2 to 6, following the synthetic scheme of steps 9 to 13 in Example 1, yielded the target compound 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamitoyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f). LC-MS (ESI) calculation for C 32 H 41 FN6O6[M+H] + m / z 625.3, found 625.4. Step 7: N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (Compound 5)

[0122] At room temperature, 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.015 g, 0.024 mmol) and N-methylcyclopropylamine (0.006 g, 0.048 mmol) were dissolved in N,N-dimethylformamide (3 mL), and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.018 g, 0.048 mmol) and N,N-diisopropylethylamine (0.019 g, 0.14 mmol) were added. The reaction mixture was allowed to react at room temperature for 16 hours, after which the reaction was terminated. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The crude product was purified by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamitoyl)phenyl)-1-propamidopropyl)-N-methyloxazol-5-carboxamide (compound 5). LCMS (ESI) calcd for C 36 H 48 FN7O5[M+H] + m / z 678.4, found 678.5. 1H NMR (400MHz, DMSO-d6) δ9.80(s,1H),8.58(d,J=8.6Hz,1H),8.42(d,J=8.2Hz,1H),7.66(t,J=8.3Hz,1H),7.57(s,1H),7.4 8(d,J=1.9Hz,1H),7.09(d,J=12.1Hz,1H),6.96-6.89(m,2H),5.45-5.33(m,1H),5.19(t,J=9.0Hz,1H),4.49(t,J=8.1Hz,1 H),2.95-2.86(m,4H),2.26-2.08(m,2H),1.87-1.74(m,2H),1.73-1.57(m,3H),1.35(dd,J=8.4,6.7Hz,6H),1.32-1.22(m, 5H),1.22-1.13(m,1H),1.12-1.02(m,1H),0.98(t,J=7.6Hz,3H),0.92-0.82(m,5H),0.79-0.66(m,2H),0.54-0.47(m,2H).

[0123] Example 6: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-((S)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 6)

[0124]

[0125] Step 1: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-((S)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 6)

[0126] (S)-3-fluoropyrrolidine hydrochloride (0.005 g, 0.042 mmol) and 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.015 g, 0.021 mmol) were dissolved in N,N-dimethylformamide (1 mL) at room temperature. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.016 g, 0.042 mmol) and N,N-diisopropylethylamine (0.016 g, 0.127 mmol) were added sequentially with stirring. The reaction mixture was then allowed to react at room temperature for 16 hours until the reaction was terminated. The reaction solution was diluted with water and extracted with ethyl acetate. The solutions were combined, dried, and the organic phase was concentrated. The crude product was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((S)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 6). LC-MS (ESI) calcd for C 36 H 47 F2N7O5[M+H] + m / z 696.4, found696.6. 1 HNMR(400MHz, DMSO-d6)δ9.82(d,J=8.2Hz,1H),8.59(t,J=7.9Hz,1H),8.47-8.37(m,1H),7.70-7.62(m,1H),7.60(d ,J=7.8Hz,1H),7.48(d,J=1.8Hz,1H),7.13(d,J=11.7Hz,1H),6.96-6.90(m,2H),5.47-5.26(m,2H),5.24-5.17(m,1 H),4.52-4.43(m,1H),3.88-3.52(m,4H),3.45-3.35(m,2H),2.28-1.99(m,4H),1.85-1.75(m,2H),1.72-1.60(m,3H ),1.37-1.33(m,6H),1.30-1.26(m,3H),1.23-1.15(m,1H),1.11-1.03(m,1H),1.01-0.96(m,3H),0.91-0.84(m,5H).

[0127] Example 7: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((R)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 9)

[0128]

[0129] Step 1: N-((S)-2-((2-fluoro-4-((1R,2S)-1-(5-(((R)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 9)

[0130] At room temperature, 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.015 g, 0.024 mmol) and (R)-3-fluoropyrrolidine hydrochloride (0.006 g, 0.048 mmol) were dissolved in N,N-dimethylformamide (3 mL). 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.018 g, 0.048 mmol) and N,N-diisopropylethylamine (0.018 g, 0.14 mmol) were added sequentially with stirring. The reaction mixture was then allowed to react at room temperature for 16 hours before termination. The reaction mixture was diluted with water and extracted with ethyl acetate. The samples were combined, dried, and the organic phase was concentrated. The crude product was purified 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-(5-(((R)-3-fluoropyrrolidine-1-carbonyl)oxazol-2-yl)-1-propamidopropyl-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 9). LCMS(ESI)calcd for C 36 H 47 F2N7O5[M+H] + m / z 696.4, found 696.5. 1H NMR (400MHz, DMSO-d6) δ9.91-9.66(m,1H),8.73-8.50(m,1H),8.41(s,1H),7.75-7.56(m,2H),7.48(s ,1H),7.14(t,J=12.0Hz,1H),7.00-6.85(m,2H),5.51-5.22(m,2H),5.21-5.11(m,1H),4.56-4.40(m, 1H),3.90-3.48(m,4H),2.29-2.04(m,4H),1.87-1.74(m,2H),1.74-1.59(m,3H),1.45-1.32(m,6H),1 .32-1.21(m,5H),1.20-1.13(m,1H),1.10-1.03(m,1H),0.98(t,J=7.6Hz,3H),0.85(d,J=5.1Hz,5H).

[0131] Example 8: N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamitoyl)phenyl)-1-propamidopropyl)-N-(methyl-d3)oxazol-5-carboxamide (Compound 11)

[0132]

[0133] Step 1: Cyclopropyl (methyl-d3)carbamate tert-butyl ester (11b)

[0134] Cyclopropyl tert-butyl carbamate (11a) (0.30 g, 1.90 mmol) was dissolved in anhydrous N,N-dimethylformamide (5 mL) at room temperature and under nitrogen protection. Sodium hydride (60%, 0.12 g, 2.86 mmol) was added with stirring at 0 °C. After reacting at room temperature for 15 min, the mixture was cooled to 0 °C, and deuterated iodomethane (0.55 g, 3.81 mmol) was slowly added. The reaction was then terminated at room temperature and under nitrogen protection for 16 h. The reaction mixture was diluted with water and extracted with ethyl acetate. The combined solutions were dried, the organic phases were concentrated, and the crude product was separated by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 20 / 1) to give cyclopropyl(methyl-d3) tert-butyl carbamate (11b). 1 H NMR (400MHz, DMSO-d6) δ2.49-2.41(m,1H),1.39(s,9H),0.66-0.62(m,2H),0.58-0.48(m,2H).

[0135] Step 2: N-(methyl-d3)cyclopropylamine (11c)

[0136] Cyclopropyl (methyl-d3)carbamate tert-butyl ester (11b) (0.12 g, 0.69 mmol) was dissolved in dichloromethane (2 mL) at 0 °C. The mixture was stirred under nitrogen protection, and a hydrogen chloride / 1,4-dioxane solution (4 M, 0.86 mL) was slowly added dropwise. The reaction was then terminated after 2 hours at room temperature under nitrogen protection. The reaction solution was concentrated to give the target product N-(methyl-d3)cyclopropylamine (11c). 1 HNMR(400MHz,DMSO-d6)δ9.22(s,2H),2.65(s,1H),0.92-0.81(m,2H),0.77-0.66(m,2H).

[0137] Step 3: N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamitoyl)phenyl)-1-propamidopropyl)-N-(methyl-d3)oxazol-5-carboxamide (Compound 11)

[0138] At room temperature, N-(methyl-d3)cyclopropylamine (11c) (0.004 g, crude) and 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamityl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.02 g, 0.03 mmol) were dissolved in N,N-dimethylformamide (1 mL). Under nitrogen protection at 0 °C and with stirring, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.018 g, 0.05 mmol) and N,N-diisopropylethylamine (0.021 g, 0.16 mmol) were added. The reaction was terminated after 1 hour under nitrogen protection. The reaction mixture was diluted with water and extracted with ethyl acetate. The samples were combined, dried, and the organic phases were concentrated. The residue was separated by preparative high-performance liquid chromatography (mobile phase: 0.1% formic acid aqueous solution / acetonitrile) to yield N-cyclopropyl-2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamitoyl)phenyl)-1-propamidopropyl)-N-(methyl-d3)oxazol-5-carboxamide (compound 11). LC-MS (ESI) calcd for C 36 H 45 D3FN7O5[M+H] + m / z 681.4, found 681.4. 1H NMR (400MHz, DMSO-d6) δ9.81(s,1H),8.59(d,J=8.7Hz,1H),8.43(d,J=8.2Hz,1H),7.66(t,J=8.4Hz,1H),7.57(s,1H),7.48(d ,J=1.9Hz,1H),7.09(d,J=10.2Hz,1H),6.95-6.90(m,2H),5.37(dd,J=13.1,6.5Hz,1H),5.19(t,J=9.1Hz,1H),4.48(t,J=8.4H z,1H),2.89(s,1H),2.17(ddt,J=22.4,14.9,7.5Hz,2H),1.84-1.57(m,5H),1.35(dd,J=8.4,6.7Hz,6H),1.27(d,J=7.0Hz,4H) ,1.20(d,J=10.5Hz,1H),1.06(d,J=10.3Hz,1H),0.98(t,J=7.6Hz,3H),0.85(d,J=6.5Hz,5H),0.77-0.64(m,2H),0.51(s,2H).

[0139] Example 9: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-(3,3,4,4-tetrafluoropyrrolidine-1-carbonyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 12)

[0140]

[0141] Step 1: N-((S)-2-((2-fluoro-4-((1R,2S)-1-propamido-1-(5-(3,3,4,4-tetrafluoropyrrolidine-1-carbonyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1R,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (Compound 12)

[0142] 3,3,4,4-Tetrafluoropyrrolidine hydrochloride (0.008 g, 0.042 mmol) and 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.015 g, 0.021 mmol) were dissolved in N,N-dimethylformamide (1 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N',N′-tetramethylurea hexafluorophosphate (0.016 g, 0.042 mmol) and N,N-diisopropylethylamine (0.016 g, 0.127 mmol) were then added sequentially. The reaction mixture was then allowed to react at room temperature for 16 hours before termination. The reaction solution was diluted with water and extracted with ethyl acetate. The solutions were combined, dried, and the organic phase was concentrated. The reaction solution was then 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-tetrafluoropyrrolidine-1-carbonyl)oxazol-2-yl)prop-2-yl)phenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (compound 12). LC-MS (ESI) calcd for C 36 H 44 F5N7O5[M+H] + m / z 750.3, found 750.4. 1 H NMR (400MHz, DMSO-d6) δ9.80(s,1H),8.58(d,J=8.6Hz,1H),8.40(d,J=8.1Hz,1H),7.72(s,1H),7.67(t,J=8.3Hz,1H),7. 48(d,J=1.9Hz,1H),7.18-7.12(m,1H),6.96(d,J=8.5Hz,1H),6.91(d,J=2.0Hz,1H),5.44-5.33(m,1H),5.25(t,J=9.0Hz ,1H),4.58-4.30(m,3H),4.28-4.08(m,2H),3.38-3.35(m,2H),2.27-2.07(m,2H),1.86-1.74(m,2H),1.72-1.58(m,3H), 1.38-1.33(m,6H),1.28(d,J=7.0Hz,3H),1.24-1.17(m,1H),1.12-1.03(m,1H),0.98(t,J=7.6Hz,3H),0.93-0.83(m,5H).

[0143] Example 10: 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamido)phenyl)-1-propamidopropyl)-N-(4-fluorophenyl)oxazol-5-carboxamide (Compound 13)

[0144]

[0145] Step 1: 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamido)phenyl)-1-propamidopropyl)-N-(4-fluorophenyl)oxazol-5-carboxamide (Compound 13)

[0146] 4-Fluoroaniline (0.005 g, 0.042 mmol) and 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-propamidopropyl)oxazol-5-carboxylic acid (5f) (0.015 g, 0.021 mmol) were dissolved in N,N-dimethylformamide (1 mL) with stirring. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (0.016 g, 0.042 mmol) and N,N-diisopropylethylamine (0.016 g, 0.127 mmol) were added sequentially. The reaction mixture was allowed to react at room temperature for 16 hours, at which point the reaction was terminated. The reaction mixture was diluted with water and extracted with ethyl acetate. The organic phases were combined, dried, and concentrated. The reaction solution was separated by high-performance liquid chromatography (HPLC) (mobile phase: ammonium bicarbonate aqueous solution / acetonitrile) to obtain the target compound 2-((1R,2S)-2-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1R,4S)-4-methylcyclohexyl)acetamido)phenyl)-1-propamidopropyl)-N-(4-fluorophenyl)oxazol-5-carboxamide (compound 13). LC-MS (ESI) calcd for C 38 H 45 F2N7O5[M+H] + m / z 718.4, found718.6. 1H NMR (400MHz, DMSO-d6) δ10.26(s,1H),9.80(s,1H),8.59(d,J=8.8Hz,1H),8.41(d,J=8.3Hz,1H),7.79(s,1H),7.70-7.64(m,3 H),7.48(d,J=1.9Hz,1H),7.19(t,J=8.9Hz,2H),7.13(d,J=12.4Hz,1H),6.97(d,J=8.4Hz,1H),6.91(d,J=2.0Hz,1H),5.40-5 .33(m,1H),5.27(t,J=8.9Hz,1H),4.47(t,J=8.3Hz,1H),3.40-3.36(m,2H),2.27-2.12(m,2H),1.84-1.73(m,2H),1.71-1.57 (m,3H),1.36-1.31(m,6H),1.28(d,J=7.0Hz,3H),1.22-1.15(m,1H),1.10-1.02(m,1H),1.01-0.96(m,3H),0.91–0.81(m,5H).

[0147] The synthesis of the following compounds is based on the preparation of compounds 1 and 5, and their characterization is shown in Table 1.

[0148] Table 1 Synthesis of Compounds

[0149]

[0150]

[0151]

[0152]

[0153]

[0154]

[0155]

[0156] To illustrate the beneficial effects of the present invention, the following experimental examples are provided.

[0157] Experimental Example 1: IL17A / A HEK-blue Cell Assay

[0158] The purpose of this experiment was to evaluate the activity of the compound at the cellular level.

[0159] 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.

[0160] 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 a 384-well plate. 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.

[0161] Calculate the RLU signal (LUMcmpd) for each aperture.

[0162] The formula for calculating the inhibition rate is as follows:

[0163] %inhibition = 100 - (Signal) cmpd -Signal Ave_PC ) / (Signal Ave_VC -Signak Ave-PC )×100.

[0164] Signal ave_pc The average signal value of the positive control.

[0165] Signal ave_vc The average signal value of the negative control.

[0166] Computing IC 50 And plot the effect-dose curve:

[0167] 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.

[0168] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))X: Logarithm of the inhibition concentration; Y: Inhibition rate.

[0169] The compounds of this invention have an IC50 of <300 nM. 50 Value, further IC 50 The value is less than 100 nM.

[0170] The results of some of the embodiments are shown in Table 2.

[0171] Table 2 Cell Inhibition Activity

[0172] Compound numbering <![CDATA[IC 50 / nM]]> IL-17modulator4 2.4 1 7.4 2 14.6 3 7.1 4 10.0 5 6.5 6 4.9 9 7.6 11 10.3 12 2.5 13 23.3

[0173] The above results indicate that the compound of the present invention has a strong inhibitory effect on IL17 cells in vitro.

[0174] Test Example 2: IL-17A / RA Binding Test

[0175] The purpose of this study was to evaluate the ability of the compound to inhibit the binding of IL-17A / RA in vitro.

[0176] IL-17 modulator 4 (CAS: 2446803-65-0) was selected as the positive control compound, and 1X reaction buffer was prepared. A 10 mM stock solution of the compound was prepared with DMSO, and 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, and 5 μL was 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.

[0177] 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.

[0178] Readings were taken using Envision HTRF mode.

[0179] Inhibition percentage = 100 - (sample signal - minimum control group average) / (maximum control group average - minimum control group average) * 100

[0180] 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.

[0181] 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: Ring A is selected from an optionally 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; 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 C 1-6 Alkyl; wherein the C 1-6 Alkyl groups are optionally substituted with one or more substituents selected from: halogens and -OR. 12 The R 12 Each is 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; X is selected from: C(R) 8 ), nitrogen, oxygen, and sulfur; Y and Z are each independently selected from: nitrogen, oxygen, and sulfur; wherein R 8 Selected from: hydrogen, fluorine and C 1-6 alkyl; R 6 and R 7 Each is independently selected from hydrogen, C 1-6 Alkyl, C 3-5 cycloalkyl, 5- to 6-membered heteroaryl and aryl; wherein the C 1-6 Alkyl, C 3-5 The cycloalkyl, 5- to 6-membered heteroaryl, and aryl groups are optionally substituted by one or more substituents selected from the following: hydrogen, halogen, C 1-3 Alkyl and C 3-5 cycloalkyl; Or R 6 and R 7 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, cyano, 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, 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 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. 1-3 Alkyl groups; preferably derived from:

4. 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.

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

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

7. The compound according to claim 1, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, R 6 and R 7 Each is independently selected from hydrogen, C 1-6 Alkyl, C 3-5 cycloalkyl, 5- to 6-membered heteroaryl and aryl; wherein the C 1-6 Alkyl, C 3-5 The cycloalkyl, 5- to 6-membered heteroaryl, and aryl groups are optionally substituted by one or more substituents selected from the following: hydrogen, halogen, C 1-3 Alkyl and C 3-5 cycloalkyl; wherein the R 6 and R 7 Each of the following is independently selected from: hydrogen, methyl, deuterated methyl, ethyl, isopropyl, cyclopropyl, cyclobutyl, C 1-3 Alkylcyclopropyl, aryl, pyridine, oxazole, isoxazole and their corresponding halogenated compounds and C 1-3 Alkane-substituted compounds; Or R 6 and R 7 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, cyano, and C. 1-6 Alkyl; wherein the C 1-6 The alkyl group is optionally substituted with one or more substituents selected from: hydrogen and halogen; wherein... Selected from:

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

9. The compound, or its stereoisomer, or its pharmaceutically acceptable salt according to any one of claims 1-8, characterized in that, The compound is described by formula II: Where C 1 C 2 C 3 and C 4 Each is independently selected from: CR 15 NR 16 N and oxygen; Wherein R 15 and R 16 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 Two of them are selected from NR 16 One of them is selected from oxygen or CR 15 The rest are CR 15 ; R 1 R 2 R 5 R 6 R 7 X, Y and Z are as described in claim 1.

10. The compound according to claim 9, or its stereoisomer, or its pharmaceutically acceptable salt, characterized in that, The compound is described by Formula III: Wherein R 16 It is methyl, ethyl, isopropyl, or fluoroethyl, R 1 R 2 It is a three-membered cycloalkyl group or R 1 R 2 It combines with the same carbon to form a six-membered cycloalkyl group, wherein the six-membered cycloalkyl group is optionally bonded to the same carbon atom. 1-3 Alkyl substitution, R 5 It is ethyl, haloethyl, or methoxymethyl, R 6 R 7 As described in claim 1.

11. The compound according to any one of claims 1-10, or its stereoisomer, or its pharmaceutically acceptable salt, wherein the compound is selected from any one of the following compounds:

12. A pharmaceutical composition, wherein, It includes the compound, stereoisomer or pharmaceutically acceptable salt thereof of any one of claims 1-11, and one or more pharmaceutically acceptable carriers, diluents or excipients.

13. Use of the compound of any one of claims 1-11 or a pharmaceutically acceptable salt thereof, or a solvate, metabolite, cocrystal or prodrug thereof, or the pharmaceutical composition of claim 12 in the preparation of a medicament for treating / preventing IL-17A-mediated diseases.

14. The use according to claim 13, wherein the disease is selected from plaque psoriasis, guttate psoriasis, inverted psoriasis, pustular psoriasis, erythrodermic psoriasis, and palmoplantar psoriasis.

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