SARS-COV-2 inhibitors with covalent modifications for treating coronavirus infections

Abstract

Provided herein are compounds, pharmaceutical compositions, and methods for treating SARS-CoV-2 infection.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 017,878, filed on April 30, 2020, U.S. Provisional Patent Application No. 63 / 059,095, filed on July 30, 2020, and U.S. Provisional Patent Application No. 63 / 112,087, filed on November 10, 2020, which are incorporated herein by reference in their entireties. Technical Field

[0003] The present disclosure relates to compounds and / or materials that are useful as potential SARS-CoV-2 inhibitors. Background Art

[0004] SARS-CoV-2 (also known as 2019-nCoV or COVID-19) first appeared in 2019. Symptoms associated with the disease include fever, myalgia, cough, dyspnea, and fatigue (Huang et al., 2020). Currently, there is no treatment available for SARS-CoV-2. However, treatments with well-known drugs such as chloroquine or investigational drugs such as remdesivir have been proposed for this disease (Colson et al., 2020; Wang et al., 2020). A mixture of the human immunodeficiency virus (HIV) drugs lopinavir / ritonavir is also being studied as a treatment for SARS-CoV-2 because they exhibit anti-coronavirus effects in vitro (Que et al., 2003; Chu et al., 2004; Chan et al., 2015; Li and De Clercq, 2020).

[0005] SARS-CoV-2 is a betacoronavirus and a member of the Coronaviridae family, which contains the largest positive-sense single-stranded RNA viruses (Cui et al., 2019). The virus contains four nonstructural proteins: papain-like (PL pro ) and 3-chymotrypsin-like (3CL pro ) protease, RNA polymerase and helicase (Zumla et al., 2016). pro and 3CL pro ) are involved in viral transcription and replication. Among the four types, 3CL proIt is believed to be primarily involved in viral replication (de Wit et al., 2016). 3CLpro hydrolyzes viral polyproteins pp1a and pp1ab during coronavirus replication to produce functional proteins. A study reported that the cysteine ​​protease 3CL of SARS-CoV-2 pro Shown to be related to SARS-CoV cysteine ​​protease 3CL pro Due to its highly conserved sequence and essential functional properties, 3CL pro It has been identified as a potential target for the development of drugs to treat SARS-CoV-2.

[0006] Because viable treatments remain elusive, there is a need for compounds and / or methods for inhibiting SARS-CoV-2 and for treating subjects infected with SARS-CoV-2. Summary of the Invention

[0007] In one aspect, provided herein is a compound comprising formula (X) or a pharmaceutically acceptable salt or solvate thereof:

[0008]

[0009] in,

[0010] B1 and B are each independently a bond, a C1-C4 alkylene, a C1-C4 heteroalkylene, or a C3-C6 cyclylene linker, wherein the alkylene, heteroalkylene, or cyclylene is optionally substituted;

[0011] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group, an acetyl cyanide group, a vinylsulfonyl group, a vinylsulfinyl group or an acryloyl group;

[0012] R3 is optionally substituted heteroaryl;

[0013] R4 is C1-C6 alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted;

[0014] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl

[0015] R 11 is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl is optionally substituted;

[0016] R 15a 、R 15b 、R 15c and R15d are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0017] wherein optionally, R 15a and R 11 Together with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring; or

[0018] wherein optionally, R 15a and R 15b Combined with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring;

[0019] R 16 is H, C1-C6 alkyl or C1-C3 haloalkyl; and

[0020] R 20 Oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, cycloalkyl sulfone, alkyl sulfone, and aryl sulfone.

[0021] In some embodiments, the compound has the structure of Formula (XA), or a pharmaceutically acceptable salt or solvate thereof:

[0022]

[0023] In another aspect, described herein is a compound having a structure of Formula (XB) or a pharmaceutically acceptable salt or solvate thereof:

[0024]

[0025] In another aspect, described herein is a compound having the structure of Formula (XI) or a pharmaceutically acceptable salt or solvate thereof:

[0026]

[0027] in,

[0028] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0029] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0030] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0031] R4 is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace;

[0032] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0033] R 11 is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0034] R 15a and R 15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0035] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, cycloalkyl sulfone, alkyl sulfone, and aryl sulfone.

[0036] In another aspect, described herein is a compound having the structure of Formula (XI) or a pharmaceutically acceptable salt or solvate thereof:

[0037]

[0038] in,

[0039] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0040] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0041] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0042] R4 is substituted cycloalkyl or optionally substituted heterocycloalkyl, wherein when substituted, each is replaced by one, two, three or four R 19 replace;

[0043] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0044] R 11 is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0045] R 15a and R 15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0046] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0047] In some embodiments, the compound has the structure of Formula (XI), or a pharmaceutically acceptable salt or solvate thereof:

[0048]

[0049] in,

[0050] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0051] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0052] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0053] R4 is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace;

[0054] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0055] R 11 is cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0056] R 15a and R15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0057] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0058] In some embodiments, the compound has the structure of Formula (XIA), or a pharmaceutically acceptable salt or solvate thereof:

[0059]

[0060] In some embodiments, B or B1 is independently a C1-C4 alkylene or a C3-C6 cyclylene linker. In some embodiments, B or B1 is independently a C2 or C3 alkylene linker. In some embodiments, B and B1 are a bond. In some embodiments, R3 is a 6-membered heteroaryl containing 1 to 3 nitrogen atoms. In some embodiments, the 6-membered heteroaryl is pyridine, pyrimidine, pyrazine, or pyridazine.

[0061] In some embodiments, the compound has the structure of Formula (XII), or a pharmaceutically acceptable salt or solvate thereof:

[0062]

[0063] wherein Y1, Y2, Y3 and Y4 are each independently CH or N, with the proviso that at least one of Y1, Y2, Y3 or Y4 is CH.

[0064] In some embodiments, Y2 is N; and Y1, Y3, and Y4 are each CH. In some embodiments, Y2 and Y4 are each N; and Y1 and Y3 are CH. In some embodiments, Y1 and Y4 are N; and Y2 and Y3 are CH. In some embodiments, Y2 and Y3 are N; and Y1 and Y4 are CH. In some embodiments, R5 is C1-C6 alkyl. In some embodiments, R5 is H.

[0065] In some embodiments, the compound has the structure of Formula (XIIA), or a pharmaceutically acceptable salt or solvate thereof:

[0066]

[0067] In some embodiments, the compound has the structure of Formula (XIIB), or a pharmaceutically acceptable salt or solvate thereof:

[0068]

[0069] In some embodiments, the compound has a stereochemical purity of at least 80%.

[0070] In some embodiments, R 15a is H; and R 15b is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy. In some embodiments, R 15a is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy; and R 15b is H. In some embodiments, R 15a and R 15c Each is H. In some embodiments, R 11 is heteroaryl, optionally substituted by one, two or three R 17 In some embodiments, the heteroaryl group is a 5-membered heteroaryl group. In some embodiments, the heteroaryl group is furan, thiophene, oxazole, thiazole, isoxazole, triazole, oxadiazole, or thiadiazole. In some embodiments, R 11 In some embodiments, R4 is optionally replaced by one, two or three R 19 In some embodiments, R4 is cycloalkyl, optionally substituted by one, two or three R19 In some embodiments, cycloalkyl is cyclobutyl, cyclopentyl, cyclohexyl, or spiro[3,3]heptanyl. In some embodiments, each R 19 are independently halogen, oxo, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 In some embodiments, each R 19 In some embodiments, R1 is a haloacetyl group, a heterocyclic acyl group, or an acryloyl group. In some embodiments, the haloacetyl group is a monosubstituted haloacetyl group or a disubstituted haloacetyl group.

[0071] In another aspect, described herein is a compound comprising the structure of Formula A, a derivative thereof, a prodrug thereof, a salt thereof, or one of its stereoisomers, or having any chirality at any chiral center, or a tautomer, polymorph, solvate, or combination thereof:

[0072]

[0073] in:

[0074] R1 is the electrophilic part;

[0075] R2, R3, R4 and R5 are substituents other than hydrogen; and

[0076] X is a heteroatom.

[0077] In another aspect, described herein is a compound comprising the structure of Formula A, a derivative thereof, a prodrug thereof, a salt thereof, or one of its stereoisomers, or having any chirality at any chiral center, or a tautomer, polymorph, solvate, or combination thereof:

[0078]

[0079] in,

[0080] R1 is the electrophilic part;

[0081] R2, R3 and R4 are substituents other than hydrogen; and

[0082] X is a heteroatom.

[0083] In some embodiments, R1 is an electrophilic moiety capable of forming a covalent bond with the cysteine ​​residue at position 145 of the SARS-CoV-2 major protease; R2 is an optionally substituted C3-C 12 alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl; R3 is optionally substituted C3-C 12 alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl; X is NH, O, S or a bond; and R4 is optionally substituted C3-C 12 Alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl.

[0084] In some embodiments, the optionally branched electrophilic moiety of R1 that can be used for covalent modification can be based on: (a) Michael acceptor (α, β-unsaturated carbonyl and sulfonyl) forms (e.g., acryloyl, vinylsulfonyl); (b) α-haloacyl (e.g., α-chloroacetyl); (c) α, β-epoxyacyl; (d) glyoxyl; (e) β, γ-diketoacyl; (f) 3,4-dioxoalkyl 3,4-dioxoalkyl; (g) 2,3-dioxoalkyl; and (h) α-ketoacyl (e.g., pyruvyl).

[0085] In another aspect, provided herein is a compound comprising a structure of one of Formula (I), (II), (III), or (IV), a derivative thereof, a prodrug thereof, a salt thereof, or a stereoisomer thereof, or having any chirality at any chiral center, or a tautomer, polymorph, solvate, or combination thereof:

[0086]

[0087] in:

[0088] R1, R2, R3, R4, R5 or R7 is a chemical moiety;

[0089] X is NH, O, S, CH2 or a bond;

[0090] each A is independently CH or N; and

[0091] B is a bond or linker.

[0092] In some embodiments, R5 and / or R6 are independently selected from H, CH3, C2H5 or CF3.

[0093] In some embodiments, Hal is a halogen, such as F, Cl, Br, or I.

[0094] In some embodiments, R2, R3, R4, R7 and / or R8 are each independently selected from H, CH3, CF3, CHF2, CH2F, C2H5, Hal, -CN or an optionally substituted moiety selected from: C3-C 12 Alkyl, C3-C 12 cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, fused heterocycle, fused aryl, fused heterocycle-aryl, spirocycle, or a combination thereof.

[0095] In some embodiments, the compound, or a pharmaceutically acceptable salt or solvate thereof, is selected from Table 1.

[0096] In another aspect, provided herein is a pharmaceutical composition comprising a compound described herein and a pharmaceutically acceptable carrier or excipient.

[0097] In another aspect, provided herein is a method of treating or preventing SARS-CoV-2 infection in a patient in need thereof, comprising administering to the patient a compound as described herein or a pharmaceutical compound as described herein. In some embodiments, the compound or pharmaceutical composition is administered to the patient until the infection is reduced or eliminated. In some embodiments, the method comprises treating one or more symptoms of SARS-CoV-2 in a patient in need thereof.

[0098] In another aspect, provided herein is an in vivo method for inhibiting a SARS-CoV-2 protease, comprising contacting the protease with a compound as described herein. In some embodiments, the compound binds to a cysteine ​​residue of the protease. In some embodiments, the compound binds reversibly or irreversibly to the cysteine ​​residue. In some embodiments, the protease is 3CL protease. In some embodiments, the cysteine ​​is cysteine ​​145 of the 3CL protease.

[0099] Other objects, features and advantages of the combinations and methods described herein will become apparent from the following detailed description. However, it should be understood that the detailed description and specific examples, while indicating specific embodiments, are given by way of illustration only, as various changes and modifications within the spirit and scope of the present disclosure will become apparent to those skilled in the art from this detailed description.

[0100] Incorporation by Reference

[0101] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0102] Various aspects of the disclosure are set forth with particularity in the appended claims.A more complete understanding of the features and advantages of the disclosure will be obtained by reference to the following detailed description and the following drawings which set forth illustrative embodiments in which the principles of the disclosure are utilized.

[0103] Figure 1 Schematic diagram showing covalent 3CL protease inhibitors for the treatment of viral infections.

[0104] Figure 2 The PK profiles of INSCoV-614(1B) upon oral, SQ, and IV administration are shown.

[0105] Figure 3 The PK profiles of INSCoV-614A(2A) upon oral, SQ, and IV administration are shown.

[0106] Figure 4 Shown with SARS-CoV-2M pro X-ray structure of complexed INSCoV-601I (1) (1.88 Angstrom resolution). DETAILED DESCRIPTION

[0107] In the following detailed description, reference is made to the accompanying drawings, which form a part thereof. Unless the context indicates otherwise, in the drawings, similar symbols generally identify similar components. The illustrative embodiments described in the detailed description, drawings, and claims are not intended to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein and illustrated in the accompanying drawings, may be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are expressly contemplated herein.

[0108] The present disclosure includes compounds and / or materials that are useful as SARS-CoV-2 inhibitors and for treating subjects infected with SARS-CoV-2. These compounds include chemical structures associated with the compound identifier INSCoV (e.g., INSCoV-number) provided herein and derivatives thereof. The compounds have various chemical structures that have been identified as inhibiting SARS-CoV-2.

[0109] Compound

[0110] In one aspect, provided herein is a compound or a tautomer, polymorph, solvate, or combination thereof, comprising a structure of Formula A*, a derivative thereof, a prodrug thereof, a salt thereof, or one of its stereoisomers, or having any chirality at any chiral center:

[0111]

[0112] in:

[0113] R1 is the electrophilic part;

[0114] R2, R3, R4 and R5 are substituents other than hydrogen; and

[0115] X is a heteroatom.

[0116] In one aspect, provided herein is a compound or a tautomer, polymorph, solvate, or combination thereof, comprising a structure of Formula A, a derivative thereof, a prodrug thereof, a salt thereof, or one of its stereoisomers, or having any chirality at any chiral center:

[0117]

[0118] in:

[0119] R1 is the electrophilic part;

[0120] R2, R3 and R4 are substituents other than hydrogen; and

[0121] X is a heteroatom.

[0122] In some embodiments, the variables are defined as follows:

[0123] R1 is an electrophilic moiety capable of forming a covalent bond with the cysteine ​​residue at position 145 of the SARS-CoV-2 main protease;

[0124] R2 is optionally substituted C3-C 12 alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl;

[0125] R3 is optionally substituted C3-C 12 alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl;

[0126] X is CH2, NH, O, S or a bond; and

[0127] R4 is optionally substituted C3-C 12 Alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycle (heterocyclic), aryl or heteroaryl.

[0128] In some embodiments, R1 is an electrophilic moiety that can be used for covalent modification. In some embodiments, R1 is:

[0129] (a) Michael acceptor (α,β-unsaturated carbonyl and sulfonyl) forms (e.g., acryloyl, vinylsulfonyl),

[0130] (b) α-haloacyl (e.g. α-chloroacetyl),

[0131] (c) α,β-epoxyacyl,

[0132] (d) glyoxyl,

[0133] (e) β,γ-diketoacyl group,

[0134] (f) 3,4-dioxoalkyl,

[0135] (g) 2,3-dioxoalkyl, and

[0136] (h) α-ketoacyl (eg, pyruvyl).

[0137] In some embodiments, the covalent modification is performed with a Michael acceptor (α,β-unsaturated carbonyl and sulfonyl) moiety (e.g., acryloyl, vinylsulfonyl). In some embodiments, the covalent modification is performed with an α-haloacyl group (e.g., α-chloroacetyl). In some embodiments, the covalent modification is performed with an α,β-epoxyacyl group. In some embodiments, the covalent modification is performed with a glyoxyl group. In some embodiments, the covalent modification is performed with a β,γ-diketoacyl group. In some embodiments, the covalent modification is performed with a 3,4-dioxoalkyl group. In some embodiments, the covalent modification is performed with a 2,3-dioxoalkyl group. In some embodiments, the covalent modification is performed with an α-ketoacyl group (e.g., pyruvoyl).

[0138] In some embodiments, the compound has a structure of Formula (I), (II), (III), or (IV), or a derivative, prodrug, salt, or stereoisomer thereof, or any chirality at any chiral center, or a tautomer, polymorph, solvate, or combination thereof:

[0139]

[0140] in:

[0141] R1, R2, R3, R4, R5 or R7 are independently a chemical moiety;

[0142] X is NH, O, S, CH2 or a bond;

[0143] Each A is independently CH or N; and

[0144] B is a bond or linker.

[0145] In some embodiments, the compound has the structure of Formula (I) or a pharmaceutically acceptable salt or solvate thereof.

[0146] In some embodiments, the compound has the structure of Formula (II) or a pharmaceutically acceptable salt or solvate thereof.

[0147] In some embodiments, the compound has the structure of Formula (III) or a pharmaceutically acceptable salt or solvate thereof.

[0148] In some embodiments, the compound has the structure of Formula (IV) or a salt or solvate thereof.

[0149] In some embodiments, each A is independently CH or N. In some embodiments, each A is independently CH. In some embodiments, each A is independently N.

[0150] In some embodiments, X is selected from NH, O, S, CH2, or a bond. In some embodiments, X is NH. In some embodiments, X is O. In some embodiments, X is S. In some embodiments, X is CH2. In some embodiments, X is a bond.

[0151] In one aspect, provided herein is a compound comprising formula (IX) or a pharmaceutically acceptable salt or solvate thereof:

[0152]

[0153] in,

[0154] B1 and B are each independently a bond, a C1-C4 alkylene, a C1-C4 heteroalkylene, or a C3-C6 cyclylene linker, wherein the alkylene, heteroalkylene, or cyclylene is optionally substituted;

[0155] R1 is

[0156] R2 is optionally substituted heteroaryl, optionally substituted aryl, optionally substituted cycloalkyl or optionally substituted heterocycloalkyl;

[0157] R3 is optionally substituted heteroaryl, optionally substituted aryl, optionally substituted cycloalkyl or optionally substituted heterocycloalkyl;

[0158] R4 is C1-C6 alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted;

[0159] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl, wherein the alkyl or haloalkyl is optionally substituted;

[0160] R 11 is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl is optionally substituted;

[0161] And R 16 is H, C1-C6 alkyl or C1-C3 haloalkyl, wherein the alkyl or haloalkyl is optionally substituted.

[0162] In some embodiments, R2 is optionally substituted cycloalkyl or heterocycloalkyl.In some embodiments, R2 is optionally substituted spirocycloalkyl or spiroheterocycloalkyl.

[0163] In some embodiments, R2 is optionally substituted aryl. In some embodiments, R2 is optionally substituted phenyl. In some embodiments, R2 is optionally substituted heteroaryl. In some embodiments, R2 is optionally substituted 5-membered heteroaryl. In some embodiments, R2 is optionally substituted 6-membered heteroaryl. In some embodiments, R2 is where R 11 、R 15a 、R 15b 、R 15c and R 15d have their meanings specified below. In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently H, amino, halogen, -CN, -OH, heteroalkyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein said heteroalkyl, alkyl, alkenyl, alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl is optionally substituted. In some embodiments, R 11 is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally substituted.

[0164] In one aspect, provided herein is a compound comprising formula (X) or a pharmaceutically acceptable salt or solvate thereof:

[0165]

[0166] in,

[0167] B1 and B are each independently a bond, a C1-C4 alkylene, a C1-C4 heteroalkylene, or a C3-C6 cyclylene linker, wherein the alkylene, heteroalkylene, or cyclylene is optionally substituted;

[0168] R1 is the electrophilic part;

[0169] R3 is optionally substituted heteroaryl;

[0170] R4 is C1-C6 alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted;

[0171] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl, wherein the alkyl or haloalkyl is optionally substituted;

[0172] R 11 is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl is optionally substituted;

[0173] R 15a 、R 15b 、R 15c and R 15d each independently represents H, amino, halogen, -CN, -OH, heteroalkyl, alkyl, alkenyl, alkynyl, haloalkyl, or alkoxy, wherein said heteroalkyl, alkyl, alkenyl, or alkynyl is optionally substituted;

[0174] wherein optionally, R 15a and R 11 Together with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring; or

[0175] wherein optionally, R 15a and R 15b Combined with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring;

[0176] And R 16 is H, C1-C6 alkyl or C1-C3 haloalkyl, wherein the alkyl or haloalkyl is optionally substituted.

[0177] In some embodiments, R 15a 、R 15b 、R 15c and R 15dEach is independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted.

[0178] In some embodiments, R 15a 、R 15b 、R 15c and R 15d are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 Substituted, where R 20 Oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, cycloalkyl sulfone, alkyl sulfone, and aryl sulfone.

[0179] In some embodiments, R1 is configured to interact with a 3CL protease. In some embodiments, R1 is an acyl group, such as a haloacetyl group, a glyoxyl group, a heterocyclic acyl group, an acetyl cyanide group, or an acryloyl group. In some embodiments, R1 is a sulfonyl group or a sulfinyl group, such as a vinylsulfonyl group or a vinylsulfinyl group.

[0180] In another aspect, provided herein is a compound having a structure of Formula (X) or a pharmaceutically acceptable salt or solvate thereof:

[0181]

[0182] in,

[0183] B1 and B are each independently a bond, a C1-C4 alkylene, a C1-C4 heteroalkylene, or a C3-C6 cyclylene linker, wherein the alkylene, heteroalkylene, or cyclylene is optionally substituted;

[0184] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group, an acetyl cyanide group, a vinylsulfonyl group, a vinylsulfinyl group or an acryloyl group;

[0185] R3 is optionally substituted heteroaryl;

[0186] R4 is C1-C6 alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted;

[0187] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0188] R 11 is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl is optionally substituted;

[0189] R 15a 、R 15b 、R 15c and R 15d are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0190] wherein optionally, R 15a and R 11 Together with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring; or

[0191] wherein optionally, R 15a and R 15b Combined with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring;

[0192] R 16 is H, C1-C6 alkyl or C1-C3 haloalkyl; and

[0193] R 20 Oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, cycloalkyl sulfone, alkyl sulfone, and aryl sulfone.

[0194] In some embodiments, the compound has the structure of Formula (XA) or a pharmaceutically acceptable salt or solvate thereof:

[0195]

[0196] In some embodiments, the compound has the structure of Formula (XB) or a pharmaceutically acceptable salt or solvate thereof:

[0197]

[0198] In some embodiments, the compound has the structure of Formula (XI) or a pharmaceutically acceptable salt or solvate thereof:

[0199]

[0200] in,

[0201] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0202] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0203] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0204] R4 is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace;

[0205] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0206] R 11is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0207] R 15a and R 15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0208] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, cycloalkyl sulfone, alkyl sulfone, and aryl sulfone.

[0209] In some embodiments, the compound has the structure of Formula (XI) or a pharmaceutically acceptable salt or solvate thereof:

[0210]

[0211] in,

[0212] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0213] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0214] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0215] R4 is substituted cycloalkyl or optionally substituted heterocycloalkyl, wherein when substituted, each is replaced by one, two, three or four R 19 replace;

[0216] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0217] R 11 is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said alkyl, aryl, cycloalkyl, heterocycloalkyl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0218] R 15a and R 15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0219] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0220] In some embodiments, the compound has the structure of Formula (XI) or a pharmaceutically acceptable salt or solvate thereof:

[0221]

[0222] in,

[0223] B is a bond, a C1-C4 alkylene or a C3-C6 cyclylene linker;

[0224] R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group or an acryloyl group;

[0225] R3 is optionally replaced by one, two or three R 18 substituted heteroaryl;

[0226] R4 is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace;

[0227] R5 is H, C1-C6 alkyl or C1-C3 haloalkyl;

[0228] R 11 is cycloalkyl, heterocycloalkyl, aryl or heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl or heteroaryl is optionally substituted by one, two or three R 17 replace;

[0229] R 15a and R 15c are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace;

[0230] Each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0231] In some embodiments, the compound has the structure of Formula (XIA) or a pharmaceutically acceptable salt or solvate thereof:

[0232]

[0233] In some embodiments, the compound has the structure of Formula (XIB) or a pharmaceutically acceptable salt or solvate thereof:

[0234]

[0235] In some embodiments, B or B1 is independently a linker. In some embodiments, B or B1 is independently a C1-C4 alkylene, C1-C4 heteroalkylene, or C3-C6 cyclylene linker.

[0236] In some embodiments, B or B1 is independently selected from:

[0237] key, wherein each A is independently CH or N; and X is NH, O, or S.

[0238] In some embodiments, B is an optionally substituted C1-C4 alkylene linker. In some embodiments, B is a C2 or C3 alkylene linker. In some embodiments, B is -CH2-, -CH2-CH2-, or -CH2-CH2-CH2-. In some embodiments, B1 is a C1-C4 alkylene linker. In some embodiments, B1 is a C2 or C3 alkylene linker. In some embodiments, B1 is -CH2-, -CH2-CH2-, or -CH2-CH2-CH2-.

[0239] In some embodiments, B is a C3-C6 cyclylene linker. In some embodiments, B is a C3, C4, C5, or C6 cyclylene linker. In some embodiments, B is In some embodiments, B1 is a C3-C6 cyclylene linker. In some embodiments, B1 is a C3, C4, C5, or C6 cyclylene linker. In some embodiments, B1 is

[0240] In some embodiments, B is a bond. In some embodiments, B1 is a bond.

[0241] In some embodiments, R3 is optionally substituted heteroaryl. In some embodiments, R3 is optionally substituted by one, two or three R 18 In some embodiments, R3 is unsubstituted heteroaryl.

[0242] In some embodiments, R3 is monocyclic or bicyclic heteroaryl.

[0243] In some embodiments, R3 is a 6-membered heteroaryl group containing 1 to 3 N atoms. In some embodiments, R3 is pyridine, pyrimidine, pyrazine, or pyridazine. In some embodiments, R 3 is pyridine. 3 In some embodiments, R 3 In some embodiments, R 3 In some embodiments, R 3 It's a pyridazine.

[0244] In some embodiments, the compound has the structure of Formula (XII) or a pharmaceutically acceptable salt or solvate thereof:

[0245]

[0246] in,

[0247] Y1, Y2, Y3 and Y4 are each independently CH or N, with the proviso that at least one of Y1, Y2, Y3 or Y4 is CH.

[0248] In some embodiments, Y2 is N; and Y1, Y3, and Y4 are each CH. In some embodiments, Y2 and Y4 are each N; and Y1 and Y3 are CH. In some embodiments, Y1 and Y4 are N; and Y2 and Y3 are CH. In some embodiments, Y2 and Y3 are N; and Y1 and Y4 are CH.

[0249] In some embodiments, R5 is H, C1-C6 alkyl, or C1-C3 haloalkyl. In some embodiments, R5 is C1-C6 alkyl. In some embodiments, R5 is H, methyl, ethyl, n-propyl, or isopropyl. In some embodiments, R5 is methyl or ethyl. In some embodiments, R5 is ethyl. In some embodiments, R5 is methyl. In some embodiments, R5 is H. In some embodiments, R5 is C1-C3 haloalkyl. In some embodiments, R5 is -CF3.

[0250] In some embodiments, the compound has the structure of Formula (XIIA) or a pharmaceutically acceptable salt or solvate thereof:

[0251]

[0252] In some embodiments, the compound has the structure of Formula (XIIB) or a pharmaceutically acceptable salt or solvate thereof:

[0253]

[0254] In some embodiments, the compounds have an ee of at least 80%, 85%, 90%, or 95%. In some embodiments, the compounds have an ee of at least 80%. In some embodiments, the compounds have an ee of at least 85%. In some embodiments, the compounds have an ee of at least 90%. In some embodiments, the compounds have an ee of at least 95%.

[0255] In some embodiments, the compounds have an ee of about 80% to about 99%. In some embodiments, the compounds have an ee of about 80%, about 85%, about 90%, or about 95%. In some embodiments, the compounds have an ee of about 90%, about 91%, about 92%, about 93%, about 94%, about 96%, about 97%, about 98%, or about 99%.

[0256] In some embodiments, R 15a 、R 15b 、R 15c and R 15d are each independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein the alkyl, alkenyl or alkynyl is optionally substituted by one, two or three R 20 replace.

[0257] In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy. 15a 、R 15b 、R 15c and R 15d Each is independently C1-C6 alkyl. In some embodiments, R 15a 、R 15b 、R 15c and R 15dare each independently methyl or tert-butyl. 15a 、R 15b 、R 15c and R 15d In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently C1-C6 haloalkyl or C1-C6 alkoxy. 15a 、R 15b 、R 15c and R 15d Each is independently -OCF3. In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently -OCH3.

[0258] In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently H, -NH2, Br, F, Cl, I, -CN, -OH, -OCF3, -CF3, -CH2CF3, -OCH3, methyl, ethyl or tert-butyl. In some embodiments, R 15a 、R 15b 、R 15c and R 15d are each independently H, F, Br, Cl or I. In some embodiments, R 15a 、R 15b 、R 15c and R 15d are each independently Br. In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each is independently Cl. In some embodiments, R 15a 、R 15b 、R 15c and R 15d Each independently is F.

[0259] In some embodiments, R 15a is H. In some embodiments, R 15b is H. In some embodiments, R 15c is H. In some embodiments, R 15d It’s H.

[0260] In some embodiments, R 15a and R11 Together with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring.

[0261] In some embodiments, R 15a and R 15b Together with the carbon atoms to which they are attached, they form a 5-6 membered substituted or unsubstituted ring.

[0262] In some embodiments, R 15a is H; and R 15b is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy. In some embodiments, R 15a is H; and R 15b It is -NH2, F, Br, Cl, I, -CN, -OH, -OCF3, -OCH3, -CF3, -CH2CF3, methyl, ethyl or tert-butyl.

[0263] In some embodiments, R 15a is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C6 haloalkyl or C1-C6 alkoxy; and R 15b is H. In some embodiments, R 15a is -NH2, F, Br, Cl, I, -CN, -OH, -OCF3, -OCH3, -CF3, -CH2CF3, methyl, ethyl or tert-butyl; and R 15b It’s H.

[0264] In some embodiments, R 11 R is amino, halogen, -CN, -OH, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, cycloalkyl, heterocycloalkyl, aryl, or heteroaryl, wherein each of said alkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, or heteroaryl is optionally substituted. In some embodiments, R 11 is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy, optionally substituted by one, two or three R 17 replace.

[0265] In some embodiments, R 11 It is a C1-C6 alkyl group, a C1-C6 haloalkyl group or a C1-C6 alkoxy group.

[0266] In some embodiments, R 11is -OCF3, -OCH3, methyl, ethyl or tert-butyl. In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 is a 3- to 6-membered heterocycloalkyl group containing 1-2 N, 1 O, and / or 1 S. In some embodiments, R 11 It's a halogen.

[0267] In some embodiments, R 11 It is -NH2, Br, Cl, I, F, -CN, -OH, -OCF3, -OCH3, -CF3, -CH2CF3, methyl, ethyl or tert-butyl.

[0268] In some embodiments, R 11 In some embodiments, R 11 Not tert-butyl.

[0269] In some embodiments, when R 15a With R 15c When both are H; then R 11 Not tert-butyl.

[0270] In some embodiments, R 11 is an optionally substituted heteroaryl. In some embodiments, R 11 is heteroaryl, optionally substituted by one, two or three R 17 In some embodiments, R 11 is an unsubstituted heteroaryl group.

[0271] In some embodiments, heteroaryl is a 5-membered heteroaryl. In some embodiments, R 11 is furan, thiophene, oxazole, thiazole, isoxazole, triazole, oxadiazole or thiadiazole. In some embodiments, R 11 In some embodiments, R 11 is thiophene. In some embodiments, R 11 In some embodiments, R 11 is thiazole. In some embodiments, R 11 In some embodiments, R 11 In some embodiments, R 11 It is an oxadiazole or a thiadiazole.

[0272] In some embodiments, R4 is C1-C6 alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted. 4 is aryl, heteroaryl, cycloalkyl or heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 In some embodiments, R4 is substituted cycloalkyl or optionally substituted heterocycloalkyl, wherein when substituted, each of them is replaced by one, two, three or four R 19 replace.

[0273] In some embodiments, R4 is cycloalkyl, optionally substituted by one, two, or three R 19 In some embodiments, the cycloalkyl group is a spirocycloalkyl group or a fused cycloalkyl group.

[0274] In some embodiments, R4 is a spirocycloalkyl. In some embodiments, R4 is a C5-C9 spirocycloalkyl. In some embodiments, R4 is an optionally substituted spiro[2.2]pentane. In some embodiments, R4 is an optionally substituted spiro[2.5]octane. In some embodiments, R4 is an optionally substituted spiro[3.5]nonane. In some embodiments, R4 is a bridged cycloalkyl. In some embodiments, R4 is a C7-C9 bridged cycloalkyl. In some embodiments, R4 is a fused cycloalkyl. In some embodiments, R4 is a C7-C9 fused cycloalkyl. In some embodiments, R4 is a fused cycloalkyl. In some embodiments, R4 is a 3-5 fused cycloalkyl. In some embodiments, R4 is substituted. In some embodiments, the cycloalkyl is optionally substituted with one or two halogens selected from Cl, Br, or F. In some embodiments, the cycloalkyl is substituted with two Fs. In some embodiments, cycloalkyl is cyclobutyl, cyclopentyl, cyclohexyl, or spiro[3,3]heptanyl. In some embodiments, R4 is optionally substituted cyclobutyl. In some embodiments, R4 is cyclopentyl. In some embodiments, R4 is cyclohexyl. In some embodiments, R4 is spiro[3,3]heptanyl.

[0275] In some embodiments, R4 is not unsubstituted cycloalkyl.In some embodiments, R4 is not cyclohexyl.

[0276] In some embodiments, when R 11 When it is tert-butyl, R4 is not cyclohexyl.

[0277] In some embodiments, R4 is optionally replaced by one, two or three R 19Substituted heterocycloalkyl. In some embodiments, R4 is a 3- to 7-membered heterocycloalkyl containing 1, 2 N, 1 O, or 1 S atoms, or a combination thereof.

[0278] In some embodiments, R4 is an optionally substituted heteroaryl (e.g., C5-C9 heteroaryl). In some embodiments, R4 is a monocyclic heteroaryl. In some embodiments, R4 is a fused heteroaryl. In some embodiments, R4 is an optionally substituted aryl (e.g., C6-C9 heteroaryl). 10 In some embodiments, R4 is optionally substituted phenyl. In some embodiments, R4 is optionally substituted naphthyl.

[0279] In some embodiments, each R 19 are independently halogen, oxo, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0280] In some embodiments, each R 19 are independently -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 Alkyl)2, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7In some embodiments, each R 19 are independently -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 Alkyl)2, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 In some embodiments, each R 19 are independently C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C 1-6 In some embodiments, each R 19 is independently -OCF3, -OCH3, methyl or ethyl. In some embodiments, each R 19 In some embodiments, each R 19 is independently Cl, Br, F, or I. In some embodiments, each R 19 It's F.

[0281] In some embodiments, R4 is selected from:

[0282]

[0283] In some embodiments, R4 is selected from:

[0284]

[0285] In some embodiments, R4 is In some embodiments, R4 is In some embodiments, R4 is In some embodiments, R4 is

[0286] In some embodiments, R1 may also include the aforementioned substituents, provided that the chemical moiety of R1 is an electrophilic moiety capable of forming a covalent bond with a cysteine ​​residue. In some embodiments, the bond is reversible. In some embodiments, the bond is irreversible. In some embodiments, R1 is a Michael acceptor. Specific examples of R1 include acrylamide, vinyl sulfone, α-chloroketone, α-ketoamide, or other covalent modifiers described herein.

[0287] In some embodiments, R1 is a haloacetyl group, a glyoxyl group, a heterocyclic acyl group, an acetyl cyanide group, a vinylsulfonyl group, a vinylsulfinyl group, or an acryloyl group. In some embodiments, R1 is a haloacetyl group. In some embodiments, the haloacetyl group is monosubstituted or disubstituted. In some embodiments, the haloacetyl group is monosubstituted. In some embodiments, the haloacetyl group is disubstituted. In some embodiments, R1 is acetyl chloride. In some embodiments, R1 is acetyl fluoride. In some embodiments, R1 is glyoxyl group. In some embodiments, R1 is a heterocyclic acyl group. In some embodiments, R1 is an acetyl cyanide group. In some embodiments, R1 is vinylsulfonyl group or vinylsulfinyl group. In some embodiments, R1 is acryloyl group.

[0288] In some embodiments, R1 may include one of the following:

[0289] wherein Hal1 and Hal2 are different halogens.

[0290] In some embodiments, R1 is:

[0291] wherein Hal1 and Hal2 are different halogens.

[0292] In some embodiments, Hal is a halogen such as F, Cl, Br, or I. In some embodiments, the halogen is F or Cl. In some embodiments, the halogen is F. In some embodiments, the halogen is Cl. In some embodiments, the halogen is Br. In some embodiments, the halogen is I.

[0293] In some embodiments, R1 is selected from In some embodiments, R1 is In some embodiments, R1 is In some embodiments, R1 is In some embodiments, R1 is

[0294] In some embodiments, R 16is H, C1-C6 alkyl or C1-C3 haloalkyl. 16 is C1-C6 alkyl. In some embodiments, R 16 In some embodiments, R 16 is C1-C3 haloalkyl. In some embodiments, R 16 is CF3 or CH2CF3. In some embodiments, R 16 It’s H.

[0295] In some embodiments, each R 17 、R 18 、R 19 and R 20 independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7 heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, and aryl sulfone.

[0296] In some embodiments, each R 17 、R 18 、R 19 and R 20 are independently -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 alkyl)2、-S(=O)2NH2、-S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 Alkyl)2, C 3-8 Cycloalkyl, C1-C6 heteroalkyl, C1-C6 alkoxy, C 1-6 Fluoroalkoxy, C 2-7In some embodiments, each R 17 、R 18 、R 19 and R 20 are independently C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C 1-6 In some embodiments, each R 17 、R 18 、R 19 and R 20 is independently -OCF3, -OCH3, methyl or ethyl. In some embodiments, each R 17 、R 18 、R 19 and R 20 In some embodiments, each R 17 、R 18 、R 19 and R 20 is independently Cl, Br, F, or I. In some embodiments, each R 17 、R 18 、R 19 and R 20 are independently Cl, Br or F.

[0297] In some embodiments, the R1, R2, R3, R4, R5, R6 and / or R7 substituents shown in the structure can each be individually substituted with substituents that are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, alkaryl, aralkyl, halo, hydroxyl, thiol, alkoxy, alkenyloxy, alkynyloxy, aryloxy, acyl, alkylcarbonyl, arylcarbonyl, acyloxy, alkoxycarbonyl, aryloxycarbonyl, halocarbonyl, alkylcarbonate, arylcarbonate, carboxyl, carboxylate, carbamoyl, mono(alkyl)substituted carbamoyl, di(alkyl)substituted carbamoyl, monosubstituted arylamino formyl, thiocarbamoyl, urea, cyano, isocyano, cyanooxy, isocyanato, isothiocyanato, azido, formyl, thioformyl, amino, mono(alkyl)- and di(alkyl)-substituted amino, mono(aryl)- and di(aryl)-substituted amino, alkylamide, arylamide, imino, alkylimino, arylimino, nitro, nitroso, sulfo, sulfonate ion group, alkylsulfanyl, arylsulfanyl, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, arylsulfonyl, phosphono, phosphonate ion group, phosphinate ion group, phosphino, phosphino, any substituent having or not having a heteroatom, derivatives thereof, and combinations thereof.

[0298] In some embodiments, the R1, R2, R3, R4, R5, R6 and / or R7 substituents shown in the structure can each be individually substituted with substituents that are independently hydrogen, halogen, hydroxy, alkoxy, straight chain aliphatic, branched chain aliphatic, cyclic aliphatic, substituted aliphatic, unsubstituted aliphatic, saturated aliphatic, unsaturated aliphatic, aromatic, polycyclic aromatic, substituted aromatic, heteroaromatic, amine, primary amine, secondary amine, tertiary amine, aliphatic amine, carbonyl, carboxyl, amide, ester, amino acid, peptide, polypeptide, derivatives thereof, substituted or unsubstituted, or combinations thereof, as well as other well-known chemical substituents.

[0299] In some embodiments, the R1, R2, R3, R4, R5, R6 and / or R7 substituents shown in the structure can each be individually substituted with substituents that are independently hydrogen, alkyl, alkenyl, alkynyl, aryl, alkaryl, aralkyl, halo, hydroxyl, thiol, alkoxy, alkenyloxy, alkynyloxy, aryloxy, acyl, alkylcarbonyl, arylcarbonyl, acyloxy, alkoxycarbonyl, aryloxycarbonyl, halocarbonyl, alkylcarbonate, arylcarbonate, carboxyl, carboxylate, carbamoyl, mono(alkyl)substituted carbamoyl, di(alkyl)substituted carbamoyl, monosubstituted arylcarbamoyl, thiocarbamoyl, urea, cyano The invention also includes but is not limited to the following: an alkyl group, an isocyano group, a cyanooxy group, an isocyanate group, an isothiocyanate group, an azido group, a formyl group, a thioformyl group, an amino group, a mono(alkyl)- and di(alkyl)-substituted amino group, a mono(aryl)- and di(aryl)-substituted amino group, an alkylamide group, an arylamide group, an imino group, an alkylimino group, an arylimino group, a nitro group, a nitroso group, a sulfo group, a sulfonate ion group, an alkylsulfanyl group, an arylsulfanyl group, an alkylsulfinyl group, an arylsulfinyl group, an alkylsulfonyl group, an arylsulfonyl group, a phosphono group, a phosphonate ion group, a phosphinate ion group, a phosphino group, a phosphino group, any substituent having or not having a heteroatom, any substituent including a straight chain, any substituent including a branched chain, and any substituent including a ring, derivatives thereof, and combinations thereof.

[0300] In some embodiments, the R1, R2, R3, R4, R5, R6 and / or R7 substituents shown in the structure can each be individually substituted with a substituent independently selected from any one or more of the following substituents: hydrogen, C1-C 24 Alkyl, C2-C 24 Alkenyl, C2-C 24 Alkynyl, C5-C 20 Aryl, C6-C 24 Alkaryl, C6-C 24 Aralkyl, halogen, hydroxyl, mercapto, C1-C 24 Alkoxy, C2-C 24 Alkenyloxy, C2-C24 Alkynyloxy, C5-C 20 Aryloxy, acyl (including C2-C 24 Alkylcarbonyl (—CO-alkyl) and C6-C 20 Arylcarbonyl (—CO-aryl)), acyloxy (—O-acyl), C2-C 24 Alkoxycarbonyl (—(CO)—O-alkyl), C6-C 20 Aryloxycarbonyl (—(CO)—O-aryl), halocarbonyl (—CO)—X, when X is a halo), C2-C 24 Alkyl carbonate ion group (—O—(CO)—O-alkyl), C6-C 20 Aryl carbonate ion group (—O—(CO)—O-aryl), carboxyl group (—COOH), carboxylate ion group (—COO - ), carbamoyl (—(CO)—NH2), single (C1-C 24 Alkyl) substituted carbamoyl (—(CO)—NH(C1-C 24 Alkyl)), di(C1-C 24 Alkyl) substituted carbamoyl (—(CO)—N(C1-C 24 alkyl)2), monosubstituted arylcarbamoyl (—(CO)—NH-aryl), disubstituted arylcarbamoyl (—(CO)—NH-aryl)2, thiocarbamoyl (—(CS)—NH2), mono(C1-C 24 alkyl) substituted thiocarbamoyl (—(CS)—NH(C1-C 24 Alkyl)), di(C1-C 24 alkyl) substituted thiocarbamoyl (—(CS)—N(C1-C 24 alkyl)2), monosubstituted arylthiocarbamoyl (—(CS)—NH-aryl), disubstituted arylthiocarbamoyl (—(CS)—NH-aryl)2, urea (—NH—(CO)—NH2), mono(C1-C 24 Alkyl) substituted urea group (—NH—(CO)—NH(C1-C 24 Alkyl)), di(C1-C 24 Alkyl) substituted urea group (—NH—(CO)—N(C1-C 24 alkyl)2), monosubstituted aryl urea (—NH—(CO)—NH-aryl), disubstituted aryl urea (—NH—(CO)—N-(aryl)2), cyano (—C≡N), isocyano (—N + ≡C - ), cyano group (—O—C≡N), isocyanate group (—O—N + ≡C- ), thiocyanate (—S—C≡N), isothiocyanate (—S—N + ≡C - ), azido (—N═N + ═N - ), formyl (—(CO)—H), thioformyl (—(CS)—H), amino (—NH2), mono (C1-C 24 Alkyl) and di(C1-C 24 Alkyl) substituted amino, mono (C6-C 20 aryl) and di(C6-C 20 Aryl) substituted amino, C2-C 24 Alkylamide (—NH—(CO)-alkyl), C5-C 20 Arylamide (—NH—(CO)-aryl), imino (—CR═NH, where R is hydrogen, C1-C 24 Alkyl, C5-C 20 Aryl, C6-C 24 Alkaryl, C6-C 24 Aralkyl, etc.), alkylimino (—CR=N(alkyl), where R=hydrogen, C1-C 24 alkyl, aryl, alkaryl, aralkyl, etc.), aryl imino (—CR═N(aryl), where R=hydrogen, alkyl, aryl, alkaryl, etc.), nitro (—NO2), nitroso (—NO), sulfonic acid (—SO2—OH), sulfonate ion (—SO2—O - ), C1-C 24 Alkylsulfanyl (—S-alkyl; also known as “alkylthio”), C5-C 20 Arylsulfanyl (—S-aryl; also known as “arylthio”), C1-C 24 Alkylsulfinyl (—(SO)-alkyl), C5-C 20 Arylsulfinyl (—(SO)-aryl), C1-C 24 Alkylsulfonyl (-SO2-alkyl), C5-C 20 Arylsulfonyl (-SO2-aryl), phosphonyl (-P(O)(OH)2), phosphonate ion (-P(O)(O - )2), phosphinate ion group (-P(O)(O-)), dioxophosphine group (-PO2), phosphine group (-PH2), any substituent with or without heteroatoms (such as N, O, P, S or other heteroatoms), wherein the heteroatom can replace carbon (such as heteroatoms replacing carbon in a chain or ring) or exchange in addition to it (such as heteroatoms added to a carbon chain or ring), including any substituents of a straight chain, any substituents including a branched chain and any substituents including a ring, their derivatives and combinations thereof.

[0301] In some embodiments, R2, R3, R4, R7 and / or R8 are each independently selected from H, CH3, CF3, CHF2, CH2F, C2H5, Hal, -CN or an optionally substituted moiety selected from: C3-C 12 Alkyl, C3-C 12 alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, fused heterocycle (heterocyclic), fused aryl (e.g., polyaryl), fused heterocycle-aryl, spirocycle (spiroalkyl, spiroheterocycle), or a combination thereof.

[0302] Any combination of the groups described above for the various variables is contemplated herein. Throughout the specification, groups and substituents thereof are chosen by one skilled in the art to provide stable moieties and compounds.

[0303] In some embodiments, the compounds prepared in the following examples are prepared from racemic starting materials (and / or intermediates) and separated into single enantiomers as final products or intermediates by chiral chromatography. Unless otherwise stated, it is understood that the absolute configuration of the isolated intermediates and final compounds as drawn is arbitrarily assigned and not determined.

[0304] Non-limiting examples of compounds or pharmaceutically acceptable salts or solvates described herein are presented in Table 1.

[0305] Table 1.

[0306]

[0307]

[0308]

[0309]

[0310]

[0311]

[0312]

[0313]

[0314]

[0315]

[0316]

[0317]

[0318]

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355] Other forms of compounds

[0356] On the other hand, the compounds described herein have one or more stereocenters, and each stereocenter exists independently in the R configuration or the S configuration. The compounds provided herein include all diastereomeric forms, enantiomeric forms and epimeric forms and their appropriate mixtures. The compounds and methods provided herein include all cis (cis), trans (trans), cis (syn), anti (anti), trans (E) and cis (Z) isomers and their appropriate mixtures. In certain embodiments, the compounds described herein are prepared into their single stereoisomers by the following manner: reacting the racemic mixture of the compound with an optically active resolving agent to form a pair of diastereomeric compounds / salts, separating the diastereoisomers, and recovering the optically pure enantiomers. In some embodiments, the covalent diastereomeric derivatives of the compounds described herein are used to separate enantiomers. In another embodiment, based on solubility differences, diastereoisomers are separated by separation / resolving techniques. In other embodiments, the separation of stereoisomers is carried out by chromatography or by forming diastereomeric salts and separating by recrystallization or chromatography or any combination thereof. Jean Jacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates and Resolutions”, John Wiley And Sons, Inc., 1981. In one aspect, stereoisomers are obtained by stereoselective synthesis.

[0357] In some embodiments, the compounds described herein are prepared as prodrugs. "Prodrug" refers to an agent that is converted into a parent drug in vivo. Prodrugs are often useful because, in some cases, they can be more easily administered than the parent drug. They can, for example, be bioavailable by oral administration, whereas the parent drug cannot. Prodrugs can also have improved solubility in pharmaceutical compositions compared to the parent drug. In some embodiments, the design of the prodrug increases effective water solubility. Without limitation, an example of a prodrug is a compound described herein that is administered in the form of an ester ("prodrug") to facilitate transport across the cell membrane (in the cell membrane, water solubility is detrimental to mobility), but then once inside the cell (inside the cell, water solubility is beneficial), it is metabolically hydrolyzed into a carboxylic acid, i.e., an active entity. Another example of a prodrug can be a short peptide (polyamino acid) bonded to an acid group, wherein the peptide is metabolized to reveal the active portion. In certain embodiments, after in vivo administration, the prodrug is chemically converted into the biologically active, pharmaceutically active, or therapeutically active form of the compound. In certain embodiments, a prodrug is enzymatically metabolized by one or more steps or processes to the biologically active, pharmaceutically active, or therapeutically active form of the compound.

[0358] In one aspect, prodrugs are designed to alter the metabolic stability or transport characteristics of a drug, mask side effects or toxicity, improve the flavor of a drug, or alter other characteristics or properties of a drug. By virtue of an understanding of in vivo pharmacokinetic, pharmacodynamic processes, and drug metabolism, once a pharmaceutically active compound is known, it is possible to design a prodrug of the compound. (See, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pp. 388-392; Silverman (1992), The Organic Chemistry of Drug Design and Drug Action, Academic Press, Inc., San Diego, pp. 352-401, Rooseboom et al., Pharmacological Reviews, 56:53-102, 2004; Aesop Cho, “Recent Advances in Oral Prodrug Discovery”, Annual Reports in Medicinal Chemistry, Vol. 41, 395-407, 2006; T. Higuchi and V. Stella, Pro-drugs as Novel Delivery Systems, Vol. 14, ACS Symposium Series).

[0359] In some cases, some of the compounds described herein may be derivatives of another or prodrugs of the active compound.

[0360] In some embodiments, sites on the aromatic ring portion of the compounds described herein are susceptible to various metabolic reactions. Therefore, incorporating appropriate substituents on the aromatic ring structure will reduce, minimize, or eliminate this metabolic pathway. In specific embodiments, by way of example only, suitable substituents to reduce or eliminate the susceptibility of the aromatic ring to metabolic reactions are halogens or alkyl groups.

[0361] In another embodiment, the compounds described herein are isotopically labeled (e.g., with a radioisotope) or labeled by another means including, but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

[0362] The compounds described herein include isotopically labeled compounds, which are identical to those depicted in the various formulas and structures provided herein, except that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, fluorine, and chlorine, such as 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 35 S. 18 F and 36 In one aspect, an isotopically labeled compound as described herein, for example, one into which a radioactive isotope such as 3 H and 14 C, are useful in drug and / or substrate tissue distribution assays. In one aspect, substitution with isotopes such as deuterium affords certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements.

[0363] In certain embodiments, for some or all 1 H atoms, in the compounds disclosed herein 2 The abundance of H atoms is enriched. In some embodiments, the compounds disclosed herein contain one deuterium atom. In another embodiment, the compounds disclosed herein contain two deuterium atoms. In another embodiment, the compounds disclosed herein contain three deuterium atoms. In another embodiment, the compounds disclosed herein contain four deuterium atoms. In another embodiment, the compounds disclosed herein contain five deuterium atoms. In another embodiment, the compounds disclosed herein contain six deuterium atoms. In another embodiment, the compounds disclosed herein contain more than six deuterium atoms. In another embodiment, the compounds disclosed herein are completely substituted with deuterium atoms and do not contain non-exchangeable 1 H hydrogen atom. In some embodiments, the level of deuterium incorporation is determined by the synthetic method in which deuterated building blocks are used as starting materials.

[0364] In additional or additional embodiments, the compounds described herein, upon administration to an organism in need thereof, are metabolized to produce metabolites, which are then used to produce a desired effect, including a desired therapeutic effect.

[0365] As used herein, "pharmaceutically acceptable" refers to materials such as carriers or diluents that do not abrogate the biological activity or properties of the compound and are relatively nontoxic, i.e., the material can be administered to a subject without causing undesirable biological effects or interacting in a deleterious manner with any component of the composition in which it is contained.

[0366] The term "pharmaceutically acceptable salt" refers to a formulation of a compound that does not cause significant irritation to an organism to which it is administered and does not abrogate the biological activity and properties of the compound. In some embodiments, pharmaceutically acceptable salts are obtained by reacting a compound described herein with an acid. Pharmaceutically acceptable salts are also obtained by reacting a compound described herein with a base to form a salt.

[0367] The compounds described herein can be formed into pharmaceutically acceptable salts and / or used in the form of pharmaceutically acceptable salts. The types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, which are prepared by reacting the free base form of the compound with a pharmaceutically acceptable inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, metaphosphoric acid, or an organic acid such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, trifluoroacetic acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, methanesulfonic acid, methylbenzene ... (2) Salts formed when an acidic proton present in the parent compound is replaced by a metal ion such as an alkali metal ion (e.g., lithium, sodium, potassium), an alkaline earth metal ion (e.g., magnesium or calcium), or an aluminum ion. In some cases, the compounds described herein can be coordinated with an organic base such as, but not limited to, ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucosamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. In other cases, the compounds described herein can form salts with amino acids such as, but not limited to, arginine, lysine, and the like. Acceptable inorganic bases for forming salts with compounds comprising acidic protons include, but are not limited to, aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.

[0368] Examples of pharmaceutically acceptable salts include those prepared by reaction of the compounds described herein with mineral acids, organic acids, or inorganic bases, such salts include acetates, acrylates, adipates, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bromide, butyrate, butyne-1,4-dioate, camphorate, camphorsulfonate, hexanoate, octanoate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, decanoate, digluconate, dihydrogen phosphate, dinitrobenzoate, dodecyl sulfate, ethanesulfonate, formate, fumarate, gluconate heptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexyne-1,6-dioate, hydroxybenzoate, gamma-hydroxybutyrate salt, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate, metaphosphate, methanesulfonate, methoxybenzoate, methylbenzoate, monohydrogen phosphate, 1-naphthalenesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, succinate, suberate, sebacate, sulfonate, tartrate, thiocyanate, toluenesulfonate, undecanoate, and xylenesulfonate.

[0369] In addition, the compounds described herein can be prepared as pharmaceutically acceptable salts by reacting the free base form of the compound with a pharmaceutically acceptable inorganic or organic acid, including but not limited to inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; and organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, p-toluenesulfonic acid, tartaric acid, trifluoroacetic acid, citric acid, Benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, arylsulfonic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 2-naphthalenesulfonic acid, 4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid, glucoheptonic acid, 4,4'-methylenebis-(3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid.

[0370] In some embodiments, compounds described herein that contain free acid groups are reacted with a suitable base of a pharmaceutically acceptable metal cation, such as hydroxide, carbonate, bicarbonate, or sulfate; with ammonia; or with a pharmaceutically acceptable organic primary, secondary, tertiary, or quaternary amine. Representative salts include alkali metal or alkaline earth metal salts, such as lithium, sodium, potassium, calcium, and magnesium and aluminum salts. Illustrative examples of bases include sodium hydroxide, potassium hydroxide, choline hydroxide, sodium carbonate, N + (C 1-4 Alkyl)4, etc.

[0371] Representative organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, and the like. It should be understood that the compounds described herein also include quaternization of any basic nitrogen-containing groups they contain. In some embodiments, water- or oil-soluble or dispersible products are obtained by such quaternization.

[0372] It should be understood that reference to pharmaceutically acceptable salts includes solvent addition forms, particularly solvates. Solvates contain stoichiometric or non-stoichiometric amounts of solvent and can be formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol, etc. When the solvent is water, hydrates are formed, or when the solvent is alcohol, alcoholates are formed. Solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. In addition, the compounds provided herein can exist in unsolvated forms as well as solvated forms. In general, for the purposes of the compounds and methods provided herein, solvated forms are considered equivalent to unsolvated forms.

[0373] In some embodiments, the compounds described herein exist in the form of solvates. The present disclosure provides methods for treating diseases by administering such solvates. The present disclosure also provides methods for treating diseases by administering such solvates in the form of pharmaceutical compositions.

[0374] Solvates contain stoichiometric or non-stoichiometric amounts of solvents and, in some embodiments, are formed during the crystallization process with pharmaceutically acceptable solvents such as water, ethanol, and the like. When the solvent is water, hydrates are formed, or when the solvent is alcohol, alcoholates are formed. Solvates of the compounds described herein can be conveniently prepared or formed during the processes described herein. In addition, the compounds provided herein can exist in unsolvated forms as well as solvated forms. In general, for the purposes of the compounds and methods provided herein, solvated forms are considered to be equivalent to unsolvated forms. Therefore, one aspect of the present disclosure relates to hydrates and solvates of the compounds of the present disclosure as described herein and / or their pharmaceutically acceptable salts, which can be separated and characterized by methods known in the art, such as thermogravimetric analysis (TGA), TGA-mass spectrometry, TGA-infrared spectroscopy, powder X-ray diffraction (PXRD), Karl Fisher titration, high-resolution X-ray diffraction, and the like.

[0375] Treatment

[0376] In another aspect, provided herein is a method of treating or preventing SARS-CoV-2 infection in a patient in need thereof, comprising administering to the patient a compound described herein or a pharmaceutical composition comprising a compound, e.g., a compound of Formula A*, A, X, IX, XI, XII, I, II, III, or IV.

[0377] In some embodiments, a compound disclosed herein is administered prophylactically to a subject. In some embodiments, a subject is suspected of having a SARS-CoV-2 infection prior to diagnosis of the SARS-CoV-2 infection.

[0378] In some embodiments, the compounds of the present disclosure are administered to a subject until the infection is treated, inhibited, or reduced. In some embodiments, the compounds are administered to a subject until one or more symptoms of SARS-CoV-2 infection are reduced.

[0379] In another aspect, provided herein is a method of inhibiting a viral infection, the method comprising providing a compound disclosed herein to the infection so as to inhibit the viral infection. In some embodiments, the viral infection is caused by SARS-CoV-2.

[0380] In another aspect, provided herein is a method for inhibiting SARS-CoV-2 by binding to a protein thereof, the method comprising providing a compound disclosed herein to SARS-CoV-2 so as to inhibit said SARS-CoV-2. In some embodiments, SARS-CoV-2 binds to a protease on SARS-CoV-2. In some embodiments, the compounds disclosed herein bind to a cysteine ​​residue of a primary protease, thereby inhibiting SARS-CoV-2. In some embodiments, the cysteine ​​residue is at position 145 of the primary protease. In some embodiments, the protease is 3CL.

[0381] Administration and pharmaceutical compositions

[0382] The compounds described herein can be used in pharmaceutical compositions for inhibiting SARS-CoV-2 to inhibit SARS-CoV-2 infection. The compounds described herein can be formulated for administration to a subject having or suspected of having SARS-CoV-2 infection by any suitable route as described herein. The compounds described herein can be used to treat a subject by inhibiting SARS-CoV-2.

[0383] In some embodiments, a pharmaceutical composition is provided, comprising an effective amount of a compound of any embodiment of an INSCoV compound (or a pharmaceutically acceptable salt thereof) for treating a disorder; wherein the disorder is a SARS-CoV-2 infection.

[0384] An "effective amount" refers to the amount of a compound or composition required to produce the desired effect. An example of an effective amount includes an amount or dose that produces acceptable levels of toxicity and bioavailability for therapeutic (drug) uses, including but not limited to the treatment of SARS-CoV-2 (2019-nCoV) infection, also known as COVID-19.

[0385] As used herein, the term "effective amount" or "therapeutically effective amount" refers to a sufficient amount of the agent or compound administered that will alleviate to some extent one or more symptoms of the disease or disorder being treated. The result may be relief and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired change in the biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound as disclosed herein required to provide clinically significant relief of disease symptoms. An example of an effective amount includes an amount or dosage that produces acceptable toxicity and bioavailability levels for therapeutic (drug) use, including but not limited to the treatment of SARS-CoV-2 (2019-nCoV) infection known as COVID-19. "Reducing" one or more symptoms (and grammatical equivalents of this phrase) means reducing the severity or frequency of one or more, or eliminating one or more symptoms. A "prophylactic effective amount" of a drug is an amount of the drug that, when administered to a subject, will have a predetermined preventive effect, such as preventing or delaying the onset (or recurrence) of an injury, disease, lesion, or illness, or reducing the likelihood of the onset (or recurrence) of an injury, disease, lesion, or illness, or their symptoms. A complete preventive effect may not occur by administering a single dose, but may only occur after administering a series of doses. Therefore, a prophylactic effective amount may be administered in one or more administrations. As used herein, an "activity reducing amount" refers to the amount of antagonist required to reduce the activity of an enzyme relative to the absence of an antagonist. As used herein, a "function-destroying amount" refers to the amount of antagonist required to destroy the function of an enzyme or protein relative to the absence of an antagonist. The precise amount will depend on the purpose of the treatment, and will be determined by one skilled in the art using known techniques (see, e.g., Lieberman, Pharmaceutical Dosage Forms (Vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th edition, 2003, Gennaro, ed., Lippincott, Williams & Wilkins).

[0386] As used herein, a "subject" or "patient" is a mammal, such as, but not limited to, a cat, dog, rodent, or primate. Typically, the subject is a human, and preferably a human who has or is suspected of having a SARS-CoV-2 infection. The terms "subject" and "patient" are used interchangeably.

[0387] Therefore, the present technology provides pharmaceutical compositions and medicaments comprising any INSCoV compound or derivative thereof, a prodrug thereof, a salt thereof, or a stereoisomer thereof as disclosed herein, or having any chirality at any chiral center, or a tautomer, polymorph, solvate, or combination thereof, and an optional pharmaceutically acceptable carrier or one or more pharmaceutically acceptable excipients or fillers. The compositions can be used in the methods and treatments described herein. Such compositions and medicaments include a therapeutically effective amount of a compound as described herein. In some embodiments, the pharmaceutical compositions can be packaged in unit dosage form. When administered to a subject in need, the unit dosage form effectively treats SARS-CoV-2 infection.

[0388] The specific dosage can be adjusted depending on the condition of the disease; the age, weight, general health, sex and diet of the subject; the dosage interval; the route of administration; the excretion rate and the combination of drugs. Any of the above dosage forms containing an effective amount is well within the scope of routine experimentation and is therefore well within the scope of the present technology.

[0389] Those skilled in the art can easily determine the effective amount, such as by simply administering the compound of the present technology to the patient in incremental amounts until the progression of the disease / disease state is reduced or stopped. The compound of the present technology can be administered to the patient at a dosage level in the range of about 0.1 to about 1,000 mg per day. For a normal adult with a body weight of about 70 kg, a dosage in the range of about 0.01 to about 100 mg per kg of body weight per day is sufficient. However, the specific dosage used may vary, or may be adjusted as deemed appropriate by one of ordinary skill in the art. For example, the dosage may depend on many factors, including the patient's needs, the severity of the disease being treated, and the pharmacological activity of the compound used. Determining the optimal dosage for a particular patient is well known to those skilled in the art.

[0390] A variety of assays and model systems can be readily utilized to determine the therapeutic effectiveness of therapies of the present technology.

[0391] Administration may include oral administration, parenteral administration, or nasal administration. In any of these embodiments, administration may include subcutaneous injection, intravenous injection, intraperitoneal injection, or intramuscular injection. In any of these embodiments, administration may include oral administration. The methods of the present technology may also include administering conventional therapeutic agents in amounts that are potentially or synergistically effective for treating SARS-CoV-2 infection, either sequentially or in combination with one or more compounds of the present technology.

[0392] In one aspect, the compounds disclosed herein are administered to a patient in an amount or dosage suitable for therapeutic use. Typically, the unit dose of a compound comprising the present technology will vary depending on patient considerations. Such considerations include, for example, age, regimen, condition, sex, extent of disease, contraindications, concomitant therapy, and the like. Exemplary unit doses based on these considerations may also be adjusted or modified by a physician skilled in the art. For example, a unit dose for a patient comprising a compound of the present technology may range from 1×10 –4 g / kg to 1 g / kg, preferably 1×10 –3 The dosage of the compounds of the present technology may also vary from 0.01 mg / kg to 100 mg / kg, or preferably, from 0.1 mg / kg to 10 mg / kg.

[0393] In some embodiments, the compounds described herein are formulated into pharmaceutical compositions. Pharmaceutical compositions are formulated in a conventional manner using one or more pharmaceutically acceptable inactive ingredients that aid in processing the active compound into a pharmaceutically usable preparation. Appropriate formulations depend on the selected route of administration. An overview of the pharmaceutical compositions described herein can be found, for example, in: Remington: The Science and Practice of Pharmacy, 19th Edition (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, HA and Lachman, L., eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, 7th Edition (Lippincott Williams & Wilkins 1999), which are incorporated herein by reference for this disclosure.

[0394] As used herein, pharmaceutical compositions refer to mixtures of the compounds disclosed herein with other chemical components (i.e., pharmaceutically acceptable inactive ingredients), such as carriers, excipients, binders, fillers, suspending agents, flavoring agents, sweeteners, disintegrants, dispersants, surfactants, lubricants, colorants, diluents, solubilizers, wetting agents, plasticizers, stabilizers, penetration enhancers, wetting agents, defoamers, antioxidants, preservatives, or one or more combinations thereof. Pharmaceutical compositions facilitate administration of the compounds to organisms.

[0395] The pharmaceutical formulations described herein can be administered to a subject in a variety of ways by a variety of routes of administration, including but not limited to oral, parenteral (e.g., intravenous, subcutaneous, intramuscular, intramedullary injection, intrathecal, direct intraventricular, intraperitoneal, intralymphatic, intranasal injection), intranasal, buccal, topical, or transdermal routes of administration. The pharmaceutical formulations described herein include but are not limited to aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposomal dispersions, aerosols, solid dosage forms, powders, immediate release formulations, controlled release formulations, fast melt formulations, tablets, capsules, pills, delayed release formulations, extended release formulations, pulsatile release formulations, multiparticulate formulations, and mixed immediate and controlled release formulations.

[0396] In some embodiments, the compounds disclosed herein are administered orally (PO).In some embodiments, the compounds disclosed herein are administered orally in the form of tablets, capsules, or pills.

[0397] In some embodiments, the compounds disclosed herein are administered by inhalation. In some embodiments, the compounds disclosed herein are formulated for intranasal administration. Such formulations include nasal sprays, nasal mists, and the like.

[0398] In some embodiments, the compounds disclosed herein are formulated as transdermal dosage forms.

[0399] In some embodiments, the compounds disclosed herein are formulated into pharmaceutical compositions suitable for intramuscular, subcutaneous, or intravenous injection. In some embodiments, the compound is administered intramuscularly. In some embodiments, the compound is administered subcutaneously (SQ). In some embodiments, the compound is administered intravenously (IV).

[0400] In any of the above-mentioned aspects are other embodiments comprising a single administration of an effective amount of the compound, including the following other embodiments: wherein (i) the compound is administered once; (ii) the compound is administered to the subject multiple times over the span of a day; (iii) the compound is administered frequently; or (iv) the compound is administered continuously. In some embodiments, the compound is administered once a day, twice a day (BID), or three times a day (TID).

[0401] In any of the above-mentioned aspects are other embodiments comprising multiple administrations of an effective amount of the compound, including the following other embodiments: wherein (i) the compound is administered continuously or intermittently: such as in a single dose; (ii) the time between multiple administrations is every 6 hours; (iii) the compound is administered to the mammal every 8 hours; (iv) the compound is administered to the mammal every 12 hours; (v) the compound is administered to the mammal every 24 hours. In other or alternative embodiments, the method includes a drug holiday, wherein the administration of the compound is temporarily suspended, or the dose of the administered compound is temporarily reduced; at the end of the drug holiday, the administration of the compound is resumed. In one embodiment, the duration of the drug holiday varies from 2 days to 1 year.

[0402] In some embodiments, the compound is administered until SARS-CoV-2 is treated. In some embodiments, the compound is administered until one or more symptoms of SARS-CoV-2 are reduced or resolved.

[0403] definition

[0404] In the following description, certain specific details are set forth to provide a thorough understanding of the various embodiments. However, those skilled in the art will appreciate that the present invention may be practiced without these details. In other cases, well-known structures have not been shown or described in detail to avoid unnecessarily obscuring the description of the embodiments. Unless the context requires otherwise, throughout the specification and the claims that follow, the word "comprise" and variations such as "comprises" and "comprising" should be interpreted in an open, inclusive sense, that is, to mean "including, but not limited to." In addition, the headings provided herein are for convenience only and do not interpret the scope or meaning of the claimed invention.

[0405] Unless otherwise indicated, the following terms as used herein have the following meanings:

[0406] As referred to in some of the definitions provided herein, by "substituted" as in "substituted alkyl," "substituted aryl," etc., it is meant that at least one hydrogen atom bonded to a carbon (or other) atom in the alkyl, aryl or other moiety is replaced with one or more non-hydrogen substituents.

[0407] In addition, if the particular group permits, the functional groups mentioned above can be further replaced by one or more additional functional groups or by one or more hydrocarbyl moieties such as those specifically listed above. Similarly, the hydrocarbyl moieties mentioned above can be further replaced by one or more functional groups or additional hydrocarbyl moieties such as those specifically listed.

[0408] When the term "substituted" appears before a list of possible substituted groups, it is intended that the term apply to each member of that group. For example, the phrase "substituted alkyl, alkenyl, and aryl" should be interpreted as "substituted alkyl, substituted alkenyl, and substituted aryl." Similarly, when the term "heteroatom-containing" appears before a list of possible heteroatom-containing groups, it is intended that the term apply to each member of that group. For example, the phrase "heteroatom-containing alkyl, alkenyl, and aryl" should be interpreted as "heteroatom-containing alkyl, heteroatom-containing alkenyl, and heteroatom-containing aryl."

[0409] As used herein, "optionally substituted" indicates that a chemical structure may be optionally substituted by a substituent group such as defined herein. That is, when a chemical structure includes optionally substituted atoms, the atoms may or may not include optional substituent groups, and thus the chemical structure may be considered to be substituted when having substituents on the atoms, or to be unsubstituted when substituents are omitted from the atoms. A substituent group, referred to as a "substituent" or "substituent group," may be coupled (e.g., covalently) to a previously unsubstituted parent structure, wherein one or more hydrogen atoms (or other substituent groups) on the parent structure have been independently replaced by one or more substituents. A substituent is a chemical moiety added to a basic chemical structure such as a chemical scaffold. Therefore, a substituted chemical structure may have one or more substituent groups on the parent structure, such as by each substituent group coupled to the atoms of the parent structure. The substituent group that may be coupled to the parent structure may be any possible substituent group. In examples of the present technology, substituent groups (e.g., R groups) can be independently selected from alkyl, -O-alkyl (e.g., -OCH3, -OC2H5, -OC3H7, -OC4H9, etc.), -S-alkyl (e.g., -SCH3, -SC2H5, -SC3H7, -SC4H9, etc.), -NR'R", -OH, -SH, -CN, -NO2 or halogen, where R' and R" are independently H or optionally substituted alkyl. Whenever a substituent is described as "optionally substituted", the substituent may also be optionally substituted with the above substituents.

[0410] In examples of the present disclosure, the substituent groups can be independently selected from: halo, alkyl, alkenyl, alkynyl, aryl, heterocyclic, thiol, alkylthio, oxo, sulfoxy, arylthio, alkylthioalkyl, arylthioalkyl, alkylsulfonyl, alkylsulfonylalkyl, arylsulfonylalkyl, alkoxy, aryloxy, aralkyloxy, aminocarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkyl, amino, trifluoromethyl, cyano, nitro, alkylamino, arylamino, alkylaminoalkyl, arylaminoalkyl, aminoalkylamino, hydroxy, alkoxyalkyl, carboxyalkyl, alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, aralkyloxycarbonyl, carboxylic acid, sulfonic acid, sulfonyl, phosphonic acid, aryl, heteroaryl, heterocyclic and aliphatic groups. It should be understood that the substituents can be further substituted. In some cases, the term "optionally substituted" or "substituted" means that the referenced group is optionally substituted with one or more additional groups individually and independently selected from the group consisting of: D, oxo, halogen, -CN, -NH2, -NH(alkyl), -N(alkyl)2, -OH, -C02H, -C02alkyl, -C(=O)NH2, -C(=O)NH(alkyl), -C(=O)N(alkyl)2, -S(=O)2NH2, -S(=O)2NH(alkyl), -S(=O)2N(alkyl)2, alkyl, cycloalkyl, fluoroalkyl, heteroalkyl, alkoxy, fluoroalkoxy, heterocycloalkyl, aryl, heteroaryl, aryloxy, alkylthio, arylthio, alkylsulfoxide, arylsulfoxide, alkylsulfone, and arylsulfone. In some other embodiments, the optional substituents are independently selected from D, halogen, oxo, -CN, -NH2, -NH(CH3), -N(CH3)2, -OH, -CO2H, -CO2(C1-C4 alkyl), -C(=O)NH2, -C(=O)NH(C1-C4 alkyl), -C(=O)N(C1-C4 alkyl)2, -S(=O)2NH2, -S(=O)2NH(C1-C4 alkyl), -S(=O)2N(C1-C4 alkyl)2, C1-C4 alkyl, C3-C6 cycloalkyl, C1-C4 fluoroalkyl, C1-C4 heteroalkyl, C1-C4 alkoxy, C1-C4 fluoroalkoxy, -SC1-C4 alkyl, -S(=O)C1-C4 alkyl and -S(=O)2(C1-C4 alkyl). In some embodiments, the optional substituents are independently selected from D, halogen, -CN, -NH2, -OH, -NH(CH3), -N(CH3)2, -NH(cyclopropyl), -CH3, -CH2CH3, -CF3, -OCH3, and -OCF3.

[0411] The term amino refers to an overall charged or net uncharged chemical group, wherein the R group can be a substituent, such as the substituents described herein.

[0412] As used herein, the term "alkyl" or "aliphatic" refers to a branched or unbranched saturated hydrocarbon group, typically but not necessarily containing 1 to about 24 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, octyl, decyl, etc., and cycloalkyl, such as cyclopentyl, cyclohexyl, etc. Typically, but not necessarily, the alkyl group herein contains 1 to about 18 carbon atoms, or 1 to about 12 carbon atoms. The term "lower alkyl" means an alkyl group having 1 to 6 carbon atoms. Substituents identified as "C1-C6 alkyl" or "lower alkyl" contain 1 to 3 carbon atoms, and such substituents contain 1 or 2 carbon atoms (i.e., methyl and ethyl). "Substituted alkyl" refers to an alkyl group substituted with one or more substituent groups, and the terms "heteroatom-containing alkyl" and "heteroalkyl" refer to an alkyl group in which at least one carbon atom is replaced by a heteroatom, as described in more detail below. If not otherwise indicated, the terms "alkyl" and "lower alkyl" include linear, branched, cyclic, unsubstituted, substituted and / or heteroatom-containing alkyl or lower alkyl groups, respectively.

[0413] As used herein, the term "alkenyl" refers to a straight chain, branched or cyclic hydrocarbon group having 2 to about 24 carbon atoms and containing at least one double bond, such as vinyl, n-propenyl, isopropenyl, n-butenyl, isobutenyl, octenyl, decenyl, tetradecenyl, hexadecenyl, eicosenyl, tetracosenyl, etc. Typically, but not necessarily, alkenyl herein contains 2 to about 18 carbon atoms, or 2 to 12 carbon atoms. The term "lower alkenyl" means an alkenyl having 2 to 6 carbon atoms, and the special term "cycloalkenyl" means a cyclic alkenyl or having 5 to 8 carbon atoms. The term "substituted alkenyl" refers to an alkenyl substituted with one or more substituents, and the terms "heteroatom-containing alkenyl" and "heteroalkenyl" refer to an alkenyl in which at least one carbon atom is replaced by a heteroatom. If not otherwise indicated, the terms "alkenyl" and "lower alkenyl" include straight-chain, branched, cyclic, unsubstituted, substituted and / or heteroatom-containing alkenyl and lower alkenyl, respectively.

[0414] As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon group having 2 to 24 carbon atoms, such as ethynyl, n-propynyl, etc., containing at least one triple bond. Typically, but not necessarily, the alkynyl group herein contains 2 to about 18 carbon atoms, or 2 to 12 carbon atoms. The term "lower alkynyl" means an alkynyl group having 2 to 6 carbon atoms. The term "substituted alkynyl" refers to an alkynyl group substituted with one or more substituent groups, and the terms "heteroatom-containing alkynyl" and "heteroalkynyl" refer to an alkynyl group in which at least one carbon atom is replaced by a heteroatom. If not otherwise indicated, the terms "alkynyl" and "lower alkynyl" include straight, branched, unsubstituted, substituted and / or heteroatom-containing alkynyl and lower alkynyl groups, respectively.

[0415] As used herein, the term "alkoxy" means an alkyl group bound by a single terminal ether bond; that is, an "alkoxy" group can be represented as -O-alkyl, wherein the alkyl group is as defined above. "Lower alkoxy" means an alkoxy group containing 1 to 6 carbon atoms, and includes, for example, methoxy, ethoxy, n-propoxy, isopropoxy, tert-butyloxy, and the like. The substituents identified herein as "C1-C6 alkoxy" or "lower alkoxy" contain 1 to 3 carbon atoms, and such substituents contain 1 or 2 carbon atoms (i.e., methoxy and ethoxy). Unless otherwise stated, "alkoxy" may be optionally substituted, for example, by the substituent groups stated above. In some embodiments, the alkoxy group is substituted with one or more halogens.

[0416] As used herein, the term "cycloalkyl" refers to a carbon atom chain, a portion of which forms a ring. Cycloalkyl may refer to a stable partially or completely saturated monocyclic or polycyclic carbocyclic ring, which may include condensation (when condensed with an aryl or heteroaryl ring, the cycloalkyl is bonded by a non-aromatic ring atom), bridging, or a spirocyclic system. It should be understood that in the embodiment including cycloalkyl, the illustrative variations of these embodiments include low cycloalkyls, such as C3-C8 cycloalkyls, cyclopropyls, cyclohexyls, 3-ethylcyclopentyls, etc. Representative cycloalkyls include, but are not limited to, cycloalkyls having three to fifteen carbon atoms (C3-C8) 15 cycloalkyl), three to ten carbon atoms (C3-C 10In some embodiments, the cycloalkyl group is a 3- to 6-membered cycloalkyl group. In some embodiments, the cycloalkyl group is a 5- to 6-membered cycloalkyl group. Monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl or carbocycle include, for example, adamantyl, norbornyl, decahydronaphthalene, bicyclo [3.3.0] octane, bicyclo [4.3.0] nonane, cis-decalin, trans-decalin, bicyclo [2.1.1] hexane, bicyclo [2.2.1] heptane, bicyclo [2.2.2] octane, bicyclo [3.2.2] nonane and bicyclo [3.3.2] decane and 7,7-dimethyl-bicyclo [2.2.1] heptane. Partially saturated cycloalkyl includes, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl. Unless otherwise specifically stated in the specification, cycloalkyl is optionally substituted, for example, by oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc. In some embodiments, cycloalkyl is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF , -OH, -OMe, -NH or -NO . In some embodiments, cycloalkyl is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF , -OH or -OMe. In some embodiments, cycloalkyl is optionally substituted with halogen.

[0417] As used herein, term "cycloalkenyl" refers to an unsaturated carbon atom chain, a portion of which forms a ring. It should be understood that in embodiments comprising cycloalkenyl, illustrative variations of these embodiments include lower cycloalkenyls, such as C3-C8, C3-C6 cycloalkenyl.

[0418] As used herein, term "alkylene" refers to a saturated carbon atom chain, which may optionally have a branched chain. It should be understood that in the embodiments comprising alkylene, illustrative variations of these embodiments include lower alkylene groups, such as C2-C4 alkylene, methylene, ethylene, propylene, 3-methylpentylene, etc.

[0419] " heteroalkyl " refers to an alkyl group in which one or more backbone atoms of the alkyl group are selected from atoms other than carbon, such as oxygen, nitrogen (e.g., -NH-, -N (alkyl)-), sulphur or a combination thereof. Heteroalkyl is attached to the remainder of the molecule at the carbon atom of the heteroalkyl group. In one aspect, heteroalkyl is a C1-C6 heteroalkyl group, wherein the heteroalkyl group comprises 1 to 6 carbon atoms and one or more atoms other than carbon, such as oxygen, nitrogen (e.g., -NH-, -N (alkyl)-), sulphur or a combination thereof, wherein the heteroalkyl group is attached to the remainder of the molecule at the carbon atom of the heteroalkyl group. Examples of such heteroalkyl groups are, for example, -CH2OCH3, -CH2CH2OCH3, -CH2CH2OCH2CH2OCH3 or -CH(CH3)OCH3. Unless specifically stated otherwise in the specification, heteroalkyl is optionally substituted with, for example, oxo, halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl etc. In some embodiments, heteroalkyl is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF , -OH, -OMe, -NH , or -NO . In some embodiments, heteroalkyl is optionally substituted with oxo, halogen, methyl, ethyl, -CN, -CF , -OH, or -OMe. In some embodiments, heteroalkyl is optionally substituted with halogen.

[0420] As used herein, the term "heterocyclic" or "heterocycle" refers to a chain of carbon and heteroatoms, wherein the heteroatoms are selected from nitrogen, oxygen, and sulfur, and a portion of the chain (at least one heteroatom) forms a ring. The term "heterocycle" may include both "aromatic heterocycles" and "non-aromatic heterocycles." Heterocycles include 4-7 membered monocyclic rings and 8-12 membered fused rings, such as imidazolyl, thiazolyl, oxazinyl, thiazinyl, dithianyl, dioxanyl, isoxazolyl, isothiazolyl, triazolyl, furanyl, tetrahydrofuranyl, dihydrofuranyl, pyranyl, tetrazolyl, pyrazolyl, pyrazinyl, pyridazinyl, imidazolyl, pyridinyl, pyrrolyl, dihydropyrrolyl, pyrrolidinyl, piperidinyl, piperazinyl, pyrimidinyl, morpholinyl, tetrahydrothiophenyl, thienyl, azetidinyl, oxetanyl, thiirane, oxirane, aziridine, indolyl, etc. "Heterocycle" may be optionally substituted at any one or more positions capable of carrying a hydrogen atom.

[0421] Unless otherwise specifically stated in the specification, the heterocycle or heterocyclic radical can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which optionally includes fused, bridged or spirocyclic ring systems. The heteroatoms in the heterocycle or heterocyclic radical are optionally oxidized. If present, one or more nitrogen atoms are optionally quaternized. The heterocycle or heterocyclic radical can be partially or fully saturated. The heterocycle or heterocyclic radical can be attached to the rest of the molecule through any atom of one or more rings. Examples of such heterocycles or heterocyclic groups include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolinyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuranyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thimorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specifically stated in the specification, the term heterocycle or heterocyclyl includes those optionally substituted with one or more substituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralkynyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R b -OR a 、-R b -OC(O)-R a 、-R b -OC(O)-OR a 、-R b -OC(O)-N(R a )2. -R b -N(R a )2. -R b -C(O)R a 、-R b -C(O)OR a 、-R b -C(O)N(R a )2. -R b -CN、-R b -OR e -C(O)N(R a )2. -R b -N(R a )C(O)OR a 、-R b -N(Ra )C(O)R a 、-R b -N(R a )S(O) t R a (where t is 1 or 2), -R b -S(O) t R a (where t is 1 or 2), -R b -S(O) t OR a (where t is 1 or 2) and -R b -S(O) t N(R a )2 (where t is 1 or 2), where each R a R is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl) or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy or trifluoromethyl), each R b are independently a direct bond or a straight or branched alkylene or alkenylene chain, and R e is a straight or branched alkylene or alkenylene chain, and wherein each of the above substituents is unsubstituted unless otherwise indicated.

[0422] As used herein, and unless otherwise specified, the term "aryl" refers to an aromatic substituent containing a single aromatic ring or multiple aromatic rings fused together, directly connected or indirectly connected (so that different aromatic rings are bound to a common group such as a methylene or ethylene moiety). Examples of aryl groups contain 5 to 20 carbon atoms, and aryl groups contain 5 to 14 carbon atoms. Exemplary aryl groups contain one aromatic ring or two fused or connected aromatic rings, such as phenyl, naphthyl, biphenyl, diphenyl ether, diphenylamine, benzophenone, etc. "Substituted aryl" refers to an aryl moiety substituted by one or more substituent groups, and the terms "aryl containing heteroatoms" and "heteroaryl" refer to aryl substituents in which at least one carbon atom is replaced by a heteroatom, as will be described in more detail below. If not otherwise indicated, the term "aryl" includes unsubstituted, substituted and / or heteroatom-containing aromatic substituents. The term "aryl" includes monocyclic and polycyclic aromatic carbocyclic groups, each of which can be optionally substituted. The term "optionally substituted aryl" refers to an aromatic monocyclic or polycyclic ring of carbon atoms, such as phenyl, naphthyl, and the like, which may be optionally substituted with one or more independently selected substituents, such as halo, hydroxy, amino, alkyl or alkoxy, alkylsulfonyl, cyano, nitro, and the like.

[0423] The term "heteroaryl" or "aromatic heterocycle" can include substituted or unsubstituted aromatic monocyclic ring structures, in some embodiments 5 to 7 membered rings, and in some embodiments 5 to 6 membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The term "heteroaryl" can also include ring systems having one or two rings, wherein at least one ring is heteroaromatic, for example, the other rings can be cycloalkyl, cycloalkenyl, cycloalkynyl, aromatic carbocycle, heteroaryl and / or heterocycle. Heteroaryl includes, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, isoxazole, pyrazole, pyridine, pyrazine, pyridazine, indole, benzofuran, benzoxazole, benzothiazole, benzimidazole and pyrimidine.

[0424] Exemplary heteroaryl groups may comprise one or more carbon atoms and one or more ring heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur, and at least one aromatic ring. In some embodiments, the heteroaryl group is a 5- to 14-membered ring system radical comprising one to thirteen carbon atoms, one to six heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur. The heteroaryl group may be a monocyclic, bicyclic, tricyclic, or tetracyclic ring system, which may include a fused (when fused to a cycloalkyl or heterocycloalkyl ring, the heteroaryl group is bonded through an aromatic ring atom) or a bridged ring system; and the nitrogen, carbon, or sulfur atoms in the heteroaryl group may be optionally oxidized; the nitrogen atom may be optionally quaternized. In some embodiments, the heteroaryl group is a 5- to 10-membered heteroaryl group. In some embodiments, the heteroaryl group is a 5- to 6-membered heteroaryl group. Examples include, but are not limited to, azacycloheptatrienyl, acridinyl, benzimidazolyl, benzothiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepenyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinadienyl, benzopyranyl, benzopyrone, benzofuranyl, benzofuranone, benzothienyl / benzothiophenyl, benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanone, isothiazolyl, imidazolyl, oxazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolyl, isoquinolinyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoaziridine, oxazolyl, oxirane, 1-oxopyridinyl, 1-oxopyrimidinyl, 1-oxopyrazinyl, 1-oxopyridazinyl, 1-phenyl-1H-pyrrolyl , phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl and thiophenyl (i.e., thienyl). Unless otherwise specifically stated in the specification, heteroaryl is optionally substituted, for example, by halogen, amino, nitrile, nitro, hydroxyl, alkyl, alkenyl, alkynyl, haloalkyl, alkoxy, aryl, cycloalkyl, heterocycloalkyl, heteroaryl, etc. In some embodiments, heteroaryl is optionally substituted by halogen, methyl, ethyl, -CN, -CF3, -OH, -OMe, -NH2 or -NO2. In some embodiments, heteroaryl is optionally substituted with halogen, methyl, ethyl, -CN, -CF 3 , -OH or -OMe. In some embodiments, heteroaryl is optionally substituted with halogen.

[0425] It is understood that each of the alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkylene and heterocycle groups may be optionally substituted with independently selected groups such as alkyl, haloalkyl, hydroxyalkyl, aminoalkyl, carboxylic acids and derivatives thereof (including esters, amides and nitriles), hydroxy, alkoxy, acyloxy, amino, alkyl and dialkylamino, acylamino, thio, and the like, and combinations thereof.

[0426] The term "spiro" or "spirocycle" refers to a compound or moiety having one atom as the only common member of two rings.

[0427] As used herein, the term "aryloxy" refers to an aryl group bound by a single terminal ether bond, wherein "aryl" is as defined above. "Aryloxy" can be represented as -O-aryl, wherein aryl is as defined above. Examples of aryloxy groups contain 5 to 20 carbon atoms, and aryloxy groups contain 5 to 14 carbon atoms. Examples of aryloxy groups include, but are not limited to, phenoxy, o-halo-phenoxy, m-halo-phenoxy, p-halo-phenoxy, o-methoxy-phenoxy, m-methoxy-phenoxy, p-methoxy-phenoxy, 2,4-dimethoxy-phenoxy, 3,4,5-trimethoxy-phenoxy, and the like.

[0428] The term "alkaryl" refers to an aryl group having an alkyl substituent, and the term "aralkyl" refers to an alkyl group having an aryl substituent, wherein "aryl" and "alkyl" are as defined above. Examples of aralkyl groups contain 6 to 24 carbon atoms, and aralkyl groups contain 6 to 16 carbon atoms. Examples of aralkyl groups include, but are not limited to, benzyl, 2-phenyl-ethyl, 3-phenyl-propyl, 4-phenyl-butyl, 5-phenyl-pentyl, 4-phenylcyclohexyl, 4-benzylcyclohexyl, 4-phenylcyclohexylmethyl, 4-benzylcyclohexylmethyl, and the like. Alkaryl groups include, for example, p-methylphenyl, 2,4-dimethylphenyl, p-cyclohexylphenyl, 2,7-dimethylnaphthyl, 7-cyclooctylnaphthyl, 3-ethyl-cyclopenta-1,4-diene, and the like.

[0429] The term "cyclic" refers to alicyclic or aromatic substituents, which may or may not be substituted and / or contain heteroatoms, and may be monocyclic, bicyclic or polycyclic.

[0430] The terms "halo" and "halogen" are used in the conventional sense to refer to chloro, bromo, and fluoro or iodo substituents.

[0431] The term "heteroatom-containing" as in "heteroatom-containing alkyl" (also referred to as "heteroalkyl") or "heteroatom-containing aryl" (also referred to as "heteroaryl") refers to a molecule, linkage, or substituent in which one or more carbon atoms are replaced with an atom other than carbon (e.g., nitrogen, oxygen, sulfur, phosphorus, or silicon, typically nitrogen, oxygen, or sulfur). Similarly, the term "heteroalkyl" refers to an alkyl substituent containing a heteroatom, the term "heterocyclic" refers to a cyclic substituent containing a heteroatom, the terms "heteroaryl" and "heteroaromatic" refer to "aryl" and "aromatic" substituents containing a heteroatom, respectively, and so on. Examples of heteroalkyl groups include alkoxyaryl groups, alkylsulfanyl-substituted alkyl groups, N-alkylated aminoalkyl groups, and the like. Examples of heteroaryl substituents include pyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl, indolyl, pyrimidinyl, imidazolyl, 1,2,4-triazolyl, tetrazolyl, and the like, and examples of heteroatom-containing alicyclic groups are pyrrolidino, morpholino, piperazino, piperidino, and the like.

[0432] The term "hydrocarbyl" refers to a monovalent hydrocarbon radical containing 1 to about 30 carbon atoms, or 1 to about 24 carbon atoms, or 1 to about 18 carbon atoms, or about 1 to 12 carbon atoms, including straight chain, branched, cyclic, saturated and unsaturated species, such as alkyl, alkenyl, aryl, etc. "Substituted hydrocarbyl" refers to a hydrocarbyl substituted with one or more substituent groups, and the term "heteroatom-containing hydrocarbyl" refers to a hydrocarbyl in which at least one carbon atom is replaced by a heteroatom. Unless otherwise indicated, the term "hydrocarbyl" should be interpreted as including hydrocarbyl moieties that are substituted and / or contain heteroatoms.

[0433] As used herein, the term "optionally substituted" or "optionally branched" or "optionally substituted" means that the group in question is unsubstituted or substituted with one or more of the specified substituents. When the group in question is substituted with more than one substituent, the substituents may be the same or different. Additionally, when the terms "independently," "independently are," and "independently selected from" are used, it is intended that the groups in question may be the same or different. Certain terms defined herein may occur more than once in a structure, and after such occurrence, each term shall be defined independently of the other terms. In some embodiments, the term "optionally substituted" or "substituted" means that the referenced group can be substituted with one or more additional groups individually and independently selected from the group consisting of alkyl, haloalkyl, cycloalkyl, aryl, heteroaryl, heterocycloalkyl, -OH, alkoxy, aryloxy, alkylthio, arylthio, alkyl sulfoxide, aryl sulfoxide, alkyl sulfone, aryl sulfone, -CN, alkyne, C1-C6 alkylalkyne, halogen, acyl, acyloxy, -CO2H, -CO2alkyl, nitro, and amino, including mono- and disubstituted amino groups (e.g., -NH2, -NHR, -NR2), and protected derivatives thereof. In some embodiments, the optional substituents are independently selected from the group consisting of alkyl, alkoxy, haloalkyl, cycloalkyl, halogen, -CN, -NH2, -NH(CH3), -N(CH3)2, -OH, -CO2H, and -CO2alkyl. In some embodiments, the optional substituents are independently selected from fluoro, chloro, bromo, iodo, -CH 3 , -CH 2 CH 3 , -CF 3 , -OCH 3 and -OCF 3 . In some embodiments, the substituted groups are substituted by one or both of the preceding groups. In some embodiments, the optional substituents on aliphatic carbon atoms (acyclic or cyclic) include oxo (=O).

[0434] As used herein, C1-C x (or C 1-x ) including C1-C2, C1-C3...C1-C x By way of example only, a group designated as "C1-C4" indicates that there are one to four carbon atoms in the moiety, i.e., the group contains 1 carbon atom, 2 carbon atoms, 3 carbon atoms, or 4 carbon atoms. Thus, by way of example only, "C1-C4 alkyl" indicates that there are one to four carbon atoms in the alkyl group, i.e., the alkyl group is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl. Furthermore, by way of example, C0-C2 alkylene includes direct bonds, -CH2-, and -CH2CH2- linkages.

[0435] "Tautomer" refers to the movement of a proton from one atom of a molecule to another atom of the same molecule. The compounds provided herein can exist as tautomers. Tautomers are compounds that can be interconverted by the migration of a hydrogen atom, which is accompanied by a switch of a single bond and an adjacent double bond. In bonding arrangements in which tautomerism is possible, a chemical equilibrium of tautomers will exist. All tautomeric forms of the compounds disclosed herein are contemplated. The exact ratio of tautomers depends on several factors, including temperature, solvent, and pH. Some examples of tautomeric interconversions include:

[0436]

[0437] All other chemical terms are defined as known in the art.

[0438] Those skilled in the art will appreciate that for this and other processes and methods disclosed herein, the functions performed in the processes and methods may be implemented in a different order. Furthermore, the outlined steps and operations are provided as examples only, and some steps and operations may be optional, combined into fewer steps and operations, or expanded into additional steps and operations without detracting from the essence of the disclosed embodiments.

[0439] With respect to the specific embodiments described in this application as illustrations of various aspects, the present disclosure will not be limited. As will be apparent to those skilled in the art, many modifications and variations can be made without departing from the spirit and scope of the present application. In addition to those methods and apparatuses listed herein, functionally equivalent methods and apparatuses within the scope of the present disclosure will be apparent to those skilled in the art based on the previous description. Such modifications and variations are intended to fall within the scope of the appended claims. The present disclosure will be limited only by the terms of the appended claims and the full scope of equivalents to which such claims are entitled. It should be understood that the present disclosure is not limited to specific methods, reagents, compound compositions, or biological systems, which may of course vary. It should also be understood that the terms used herein are only for the purpose of describing specific embodiments and are not intended to be restrictive.

[0440] With respect to the use of substantially any plural and / or singular terms herein, those skilled in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. For clarity, the various singular / plural permutations may be expressly set forth herein.

[0441] Those skilled in the art will appreciate that, in general, the terms used herein, and particularly in the appended claims (e.g., the bodies of the appended claims), are generally intended to be "open" terms (e.g., the term "including" should be interpreted as "including but not limited to," the term "having" should be interpreted as "having at least," the term "comprising" should be interpreted as "including but not limited to," etc.). Those skilled in the art will further appreciate that if a specific number of an introduced claim recitation is intended, such intent will be explicitly recited in the claim, and in the absence of such recitation, no such intent is present. For example, as an aid to understanding, the following appended claims may contain usage of the introductory phrases "at least one" and "one or more" to introduce claim recitations. However, the use of such phrases should not be construed as implying that introducing claim recitations by the indefinite article "a / an" limits any particular claim containing such introduced claim recitation to embodiments containing only one such recitation, even when the same claim includes the introductory phrases "one or more" or "at least one" and an indefinite article such as "a / an" (e.g., "a / an" should be construed to mean "at least one" or "one or more"; this also applies to the use of definite articles to introduce claim recitations). Furthermore, even if a specific number of introduced claim recitations is explicitly recited, one skilled in the art will recognize that such recitation should be construed to mean at least the recited number (e.g., the mere recitation of "two recitations" without other modifiers means at least two recitations, or two or more recitations). Furthermore, in those instances where a convention similar to “at least one of A, B, and C, etc.” is used, generally speaking, such construction is intended in the sense that one skilled in the art would understand the convention (e.g., “a system having at least one of A, B, and C” would include but is not limited to systems having A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). In those instances where a convention similar to “at least one of A, B, or C, etc.” is used, generally speaking, such construction is intended in the sense that one skilled in the art would understand the convention (e.g., “a system having at least one of A, B, or C” would include but is not limited to systems having A alone, B alone, C alone, A and B together, A and C together, B and C together, and / or A, B, and C together, etc.). One skilled in the art would further understand that virtually any transitional word and / or phrase presenting two or more alternative terms, whether in the specification, claims, or drawings, should be understood to contemplate the possibility of including one, either, or both of the terms.For example, the phrase "A or B" should be understood to include the possibilities of "A" or "B" or "A and B."

[0442] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group.

[0443] As will be appreciated by those skilled in the art, for any and all purposes, such as providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be readily considered to fully describe and enable decomposition of the range into at least equal halves, thirds, fourths, fifths, tenths, etc. As a non-limiting example, each range discussed herein can be readily decomposed into a lower third, a middle third, and an upper third, etc. As will be appreciated by those skilled in the art, all expressions such as "at most," "at least," etc. include the recited values ​​and refer to ranges that can subsequently be decomposed into subranges as discussed above. Finally, as will be appreciated by those skilled in the art, ranges include each individual member. Thus, for example, a group having 1-3 units refers to groups having 1, 2, or 3 units. Similarly, a group having 1-5 units refers to groups having 1, 2, 3, 4, or 5 units, and so on.

[0444] As used herein, the terms "co-administered," "co-administered," and the like are intended to encompass the administration of selected therapeutic agents to a single patient, and are intended to include treatment regimens in which the agents are administered by the same or different routes of administration, or at the same or different times.

[0445] The term "pharmaceutical combination" as used herein means a product resulting from the mixing or combining of more than one active ingredient, and includes both fixed and non-fixed combinations of the active ingredients. The term "fixed combination" means that the active ingredients, e.g., a compound of formula (I) and a co-agent, are administered to a patient simultaneously in the form of a single entity or dosage. The term "non-fixed combination" means that the active ingredients, e.g., a compound of formula (I) and a co-agent, are administered to a patient simultaneously, concurrently, or sequentially in the form of separate entities with no specific intervening time limit, wherein such administration provides effective levels of the two compounds in the patient's body. The latter also applies to cocktail therapy, e.g., administration of three or more active ingredients.

[0446] As used herein, the terms "treat," "treating," and "treatment" include prophylactically and / or therapeutically alleviating, reducing, or ameliorating at least one symptom of a disease or disorder; preventing additional symptoms; inhibiting the disease or disorder, such as arresting the development of the disease or disorder; relieving the disease or disorder; causing regression of the disease or disorder; alleviating the condition caused by the disease or disorder; or halting the symptoms of the disease or disorder.

[0447] The term "regimen" refers to the dosing and timing of one or more therapies (e.g., a combination described herein or another active agent such as, for example, an anticancer agent described herein) for treating a disease, condition, or disorder described herein. A regimen may include periods of active administration and periods of rest as known in the art.

[0448] In light of the foregoing, it will be appreciated that the various embodiments of the present disclosure have been described herein for illustrative purposes, and that various modifications may be made without departing from the scope and spirit of the present disclosure. Therefore, the various embodiments disclosed herein are not intended to be limiting, with the true scope and spirit being indicated by the claims.

[0449] All references recited herein are hereby specifically incorporated by reference in their entirety: US2011 / 0269834, US2017 / 0313685; and WO 2010 / 022455.

[0450] Example

[0451] It should be understood that the following examples are intended to illustrate and not to limit the present disclosure. Various other embodiments and modifications to the foregoing descriptions and embodiments will become apparent to those skilled in the art upon reading this disclosure without departing from the spirit and scope of the present disclosure, and it is intended that all such embodiments or modifications be included within the scope of the appended claims. All publications and patents mentioned herein are hereby incorporated by reference in their entirety.

[0452] Example 1. General synthesis method

[0453] In other embodiments, starting materials and reagents used in the synthesis of compounds described herein are synthesized or obtained from commercial sources such as, but not limited to, Sigma-Aldrich, Fisher Scientific (Fisher Chemicals), and Acros Organics.

[0454] The compounds described herein and other related compounds having various substituents are synthesized using the techniques and materials described herein, as well as those recognized in the art, such as described, for example, in Fieser and Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 and Supplements (Elsevier Science Publishers, 1989); Organic Reactions, Volumes 1-40 (John Wiley and Sons, 1991), Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989), March, Advanced Organic Chemistry 4th Edition, (Wiley 1992); Carey and Sundberg, Advanced Organic Chemistry 4th Edition, Volumes A and B (Plenum 2000, 2001), and Green and Wuts, Protective Groups in Organic Chemistry 4th Edition. Synthesis 3rd edition, (Wiley 1999) (all of which are incorporated herein by reference for such disclosure). General methods for preparing compounds as disclosed herein can be derived from reactions, and the reactions can be modified by using appropriate reagents and conditions to introduce the various moieties found in the formulas provided herein. As a guide, the following synthetic methods can be utilized.

[0455] The yields reported herein refer to the purified product (unless otherwise specified). Analytical TLC was performed on Merck silica gel 60F 254 The compounds were visualized by UV light and / or stained with iodine, ninhydrin or potassium permanganate solution followed by heating. Flash column chromatography was performed on silica gel. On a plate with 5 mm DUL (double) 13 The data were recorded on a Bruker 400 MHz Avance II spectrometer with a C probe and a Bruker 400 MHz Avance III HD spectrometer with a BBFO (Broadband Fluorine Observation) probe. 1 H-NMR spectra. Chemical shifts (δ) are reported in parts per million (ppm) referenced to the peak of the deuterated solvent in which the sample was prepared. Splitting patterns are designated as s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), and br s (broad singlet).

[0456] The following solvents, reagents or scientific terms may be referred to by their abbreviations:

[0457] DCM dichloromethane

[0458] DEA Diethylamine

[0459] ETOAc or EA Ethyl acetate

[0460] EtOH

[0461] HPLC high-performance liquid chromatography

[0462] LCMS liquid chromatography-mass spectrometry

[0463] MeCN or ACN Acetonitrile

[0464] MeOH methanol

[0465] MTBE methyl tert-butyl ether

[0466] PE petroleum ether

[0467] SFC Supercritical Fluid Chromatography

[0468] THF Tetrahydrofuran

[0469] TLC thin layer chromatography

[0470] mL milliliters

[0471] mmol millimole

[0472] h hour

[0473] min

[0474] g grams

[0475] mg milligrams

[0476] eq equivalent

[0477] rt or RT Room temperature (25)

[0478] CF3CH2OH 2,2,2-trifluoromethanol

[0479] TiCl4 Titanium(V) chloride

[0480] TEA or Et3N triethylamine

[0481] IPA or i-PrOH Isopropyl alcohol

[0482] In one aspect, the compounds described herein are synthesized according to the Ugi-type reaction exemplified in Scheme 1 for the preparation of the INSCoV series.

[0483] Solution 1

[0484]

[0485] General procedure for the preparation of the INSCoV series. To a solution of 2-chloroacetic acid (1.00 eq) and isonitrile (1.00 eq) in 2,2,2-trifluoroethanol (7 mL / mmol) at 25°C was added an amine (1.00 eq) and an aldehyde (1.00 eq). The mixture was stirred at 25°C for 1 h. LC-MS showed complete consumption of the amine and a major peak with the desired mass was detected. The solvent was evaporated under reduced pressure to yield a residue. The general purification method is listed below.

[0486] Purification A: The residue was purified by preparative HPLC.

[0487] Purification B: The residue was dissolved in 10 mL of EtOAc and washed with 10 mL of water, then separated and the organic phase was dried over anhydrous Na2SO4 and filtered. The organic phase was concentrated in vacuo to give the crude product. The crude product was triturated from the solvent.

[0488] In some embodiments, the compounds prepared in the following examples are prepared from racemic starting materials (and / or intermediates) and separated into single enantiomers as final products or intermediates by chiral chromatography. Unless otherwise stated, it should be understood that the absolute configuration of the separated intermediates and final compounds as drawn is arbitrarily specified and not measured. In some embodiments, the absolute stereochemistry of enantiomers as drawn is arbitrarily specified. In some embodiments, two enantiomers are synthesized.

[0489] In some embodiments, stereochemistry is assigned based on modeling and activity data. From a modeling perspective, the R-isomer tends to bind more readily to proteases, while the S-isomer may be inactive due to poor posture. For some compounds, the chiral configuration is confirmed by X-ray studies or by chiral synthesis. For example, INSCoV-601I (1) was confirmed to be in the R-configuration by X-ray binding mode. For example, the second chiral center of the following compounds is assigned based on the chiral starting material: INSCoV-600B (1), 600B (2), 600C (1), 600C (2), 601Q, 601R, and 601S. As another example, the second chiral center of the isonitrile component from INSCoV-601Q is assigned based on the chiral starting material.

[0490] Example 2. Synthesis of INSCoV-517A, INSCoV-517(1A) and INSCoV-517A(1B)

[0491] Option 2

[0492]

[0493] Step 1: To a stirred solution of 2-amino-5-(trifluoromethoxy)benzonitrile 1 (20 g, 98.90 mmol) and pyrimidine-5-carbaldehyde 2 (11.80 g, 109 mmol) in dichloromethane (2 L) was added triethylamine (30 g, 297 mmol) and TiCl 4 (9.38 g, 49.50 mmol) at 0° C. under an inert atmosphere. The reaction mixture was stirred at room temperature for 2 h. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with ice-cold water (2 L) and extracted with dichloromethane (2×2 L). The combined organic layers were washed with brine solution (3 L), dried over Na2SO4, filtered and evaporated under reduced pressure to give crude (E)-2-((pyrimidin-5-ylmethylene)amino)-5-(trifluoromethoxy)benzonitrile 7, which was purified by flash chromatography (silica gel, 120 g SNAP) using 5% ethyl acetate / heptane eluent to give the desired product (18 g, 56%) as a light yellow solid.

[0494] 1 H NMR (400MHz, DMSO d6): δ9.38 (s, 1H), 9.30 (s, 2H), 8.87 (s, 1H), 8.10 (s, 1H), 7.88-7.86 (d, J = 8.8Hz, 1H) and 7.66-7.64 (d, J = 9.2Hz, 1H). LCMS = [M+H] + :(293.05), purity = 93%.

[0495] Step 2: To a stirred solution of 2-chloroacetic acid 5 (4.85 g, 51.3 mmol) in CF3CH2OH (50 mL) was added (E)-2-((pyrimidin-5-ylmethylene)amino)-5-(trifluoromethoxy)benzonitrile 7 (5.0 g, 17.1 mmol) and 1,1-difluoro-4-isocyanatocyclohexane 8 (4.97 g, 34.2 mmol) at room temperature under an inert atmosphere. The reaction mixture was stirred at room temperature for 48 h. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (2×250 mL). The combined organic layers were washed with brine solution (250 mL), dried over Na2SO4, filtered and evaporated under reduced pressure to give the crude product, which was purified by column chromatography (silica gel, 100-200 mesh) using 35% ethyl acetate / heptane eluent to afford 2-chloro-N-(2-cyano-4-(trifluoromethoxy)phenyl)-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)acetamide (INSCoV-517A) (350 mg, 74% purity) as a brown solid. The obtained compound was further purified by reverse phase purification to give the desired product INSCoV-517A (150 mg, 2%) as a white solid.

[0496] 1 H NMR(400MHz,DMSO d6): δ9.15-8.78(m,1H),8.47(s,2H),8.37-8.35(d,J=7.2Hz,1H),8.27-8.25(d,J=8.0Hz,1H),7.92-7.81(m,2H),6.19-6.04(m,1H), 4.23-4.05(m,2H), 3.82-3.62(m,1H), 2.02-1.95(m,1H), 1.92-1.79(m,4H), 1.71-1.64(m,1H), 1.55-1.46(m,1H) and 1.34-1.22(m,1H). LCMS=[M+H] + :(532.16), purity = 99.60%.

[0497] Step 3: Chiral HPLC purification of INSCoV-517A: INSCoV-517A (1A) and INSCoV-517A (1B). 2-Chloro-N-(2-cyano-4-(trifluoromethoxy)phenyl)-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)acetamide (INSCoV-517A, 180 mg) was purified by chiral HPLC using (column: CHIRALPAK IG (250*21) mm, 5 μm; mobile phase: Ai-PrOH (25%) and B-hexane (75%); flow mode: isocratic, loading amount: 5 mg / injection, run time: 20 min, wavelength: 234 nm, sample preparation: acetonitrile and i-PrOH) to produce INSCoV-517A(1A) (65 mg, 72%) and INSCoV-517A(1B) (80 mg, 80%) as white solids.

[0498] INSCoV-517A(1A): 1 H NMR(400MHz,DMSO d6): δ9.15-8.78(m,1H),8.48(s,2H),8.36-8.34(d,J=7.2Hz,1H),8.27-8.25(d,J=8.0Hz,1H),7.91-7.82(m,2H),6.29-6.07(m,1H), 4.23-4.05(m,2H),3.82-3.62(m,1H),2.02-1.95(m,1H),1.92-1.79(m,4H),1.71-1.64(m,1H),1.55-1.46(m,1H),1.34-1.22(m,1H). LCMS=[M+H] + :(532.16), purity = 97.20%. Chiral purity: 99.2% ee.

[0499] INSCoV-517A(1B): 1 H NMR(400MHz,DMSO d6): δ9.15-8.78(m,1H),8.46(s,2H),8.36-8.34(d,J=7.2Hz,1H),8.27-8.25(d,J=8.0Hz,1H),7.92-7.81(m,2H),6.29-6.07(m,1H), 4.23-4.05(m,2H),3.82-3.62(m,1H),2.02-1.95(m,1H),1.92-1.79(m,4H),1.71-1.64(m,1H),1.55-1.46(m,1H),1.34-1.22(m,1H). LCMS=[M+H]+ :(532.16), purity = 97.8%. Chiral purity: 99.70% ee.

[0500] Example 3. INSCoV-517C, INSCoV-517C(1), INSCoV-517C(2), INSCoV-517C(3) and Synthesis of INSCoV-517C(4)

[0501] Option 3

[0502]

[0503] Step 1: To a stirred solution of 2-chloro-2-fluoroacetic acid 6 (5.39 g, 47.9 mmol) in CF3CH2OH (50 mL) was added (E)-2-((pyrimidin-5-ylmethylene)amino)-5-(trifluoromethoxy)benzonitrile 3 (7.0 g, 47.9 mmol) and 1,1-difluoro-4-isocyanatocyclohexane 4 (6.95 g, 47.9 mmol) at room temperature under an inert atmosphere. The reaction mixture was stirred at room temperature for 48 h. After completion of the reaction (TLC monitoring), the reaction mixture was diluted with water (250 mL) and extracted with ethyl acetate (2×250 mL). The combined organic layers were washed with brine solution (300 mL), dried over Na2SO4, filtered and evaporated under reduced pressure to give the crude product, which was purified by column chromatography (silica gel, 100-200 mesh) using 25% ethyl acetate / heptane eluent to afford the desired product (1.40 g, 72% purity) as a brown solid, which was further purified by reverse phase purification to afford the desired product INSCoV-517C (mixture of diastereomers) (822 mg, 6.27%) as a white solid.

[0504] 1 H NMR(400MHz,DMSO d6): δ9.17-8.79(m,1H),8.45-8.40(m,2H),8.38-8.36(m,1H),8.30-8.17(m,1H),7.94-7.88(m,2H),6.89 -6.75(m,1H),6.24-6.00(m,1H),3.81(brs,1H),1.99-1.66(m,6H),1.48-1.36(m,1H),1.26-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 99.09%

[0505] Step 2: Diastereoisomeric separation of INSCoV-517C to give INSCoV-517C (D1) and INSCoV-517C (D2). 2-Chloro-N-(2-cyano-4-(trifluoromethoxy)phenyl)-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-2-fluoroacetamide (INSCoV-517C, 822 mg) was purified by reverse phase purification, and two diastereomers, INSCoV-517C (D1) [305 mg, 74%] and INSCoV-517C (D2) [317 mg, 77%], were isolated.

[0506] INSCoV-517C(D1): 1 H NMR(400MHz,DMSO d6): δ9.24-8.79(m,1H),8.45(s,2H),8.38-8.36(d,J=7.2Hz,1H),8.30-8.28 (d,J=7.2Hz,1H),7.95-7.92(m,2H),6.89-6.72(m,1H),6.11(s,1H),3.77(br s,1H),1.98-1.68(m,6H),1.52-1.45(m,1H),1.41-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 99.72%.

[0507] INSCoV-517C(D2): 1 H NMR(400MHz,DMSO d6): δ9.24-8.84(m,1H),8.44(s,2H),8.41-8.39(d,J=7.2Hz,1H),8.28-8.26 (d,J=7.2Hz,1H),7.95-7.88(m,2H),6.87-6.56(m,1H),6.24(s,1H),3.83(br s,1H),1.98-1.68(m,6H),1.52-1.45(m,1H),1.41-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 98.8%.

[0508] Step 3: Chiral separation of INSCoV-517C (D1): INSCoV-517C (1) and INSCoV-517C (2). Chiral HPLC purification of INSCoV-517C (D1) [305 mg] was performed using (column: CHIRALPAK IG (250*30) mm, 5 μm; mobile phase: A-EtOH (15%) and B-0.1% formic acid in hexane (85%); flow mode: isocratic, sample load: 20 mg / injection, run time: 35 min, wavelength: 230 nm, sample preparation: acetonitrile and i-PrOH) to produce INSCoV-517C (1) [88 mg, 58%] and INSCoV-517C (2) [55 mg, 35%] as white solids.

[0509] INSCoV-517C(1): 1 H NMR(400MHz,DMSO d6): δ9.22-8.88(m,1H),8.45(s,2H),8.38-8.36(d,J=7.2Hz,1H),8.30-8.28 (d,J=7.2Hz,1H),7.95-7.63(m,2H),6.89-6.72(m,1H),6.11(s,1H),3.77(br s,1H),1.98-1.87(m,6H),1.70-1.66(m,1H),1.51-1.45(m,1H). LCMS=[M+H] + :(550.18), purity = 99.8%. Chiral purity: 99.4% ee.

[0510] INSCoV-517C(2): 1 H NMR(400MHz,DMSO d6): δ9.17-8.79(m,1H),8.45(s,2H),8.38-8.36(d,J=7.2Hz,1H),8.30-8.28 (d,J=7.2Hz,1H),7.95-7.88(m,2H),6.89-6.72(m,1H),6.11(s,1H),3.77(br s,1H),1.98-1.68(m,6H),1.52-1.45(m,1H),1.41-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 98.8%. Chiral purity: 99.7% ee.

[0511] Step 4: Chiral separation of INSCoV-517C (D2): INSCoV-517C (3) and INSCoV-517C (4). Chiral HPLC purification of INSCoV-517C (D2) [317 mg] was performed using (column: CHIRALPAK IG (250*30) mm, 5 μm; mobile phase: A-EtOH (20%) and B-0.1% formic acid in hexane (80%); flow mode: isocratic, sample load: 20 mg / injection, run time: 20 min, wavelength: 230 nm, sample preparation: acetonitrile and i-PrOH) to produce INSCoV-517C (3) [110 mg, 70%] and INSCoV-517C (4) [105 mg, 66%] as white solids.

[0512] INSCoV-517C(3): 1 H NMR(400MHz,DMSO d6): δ9.17-8.84(m,1H),8.44(s,2H),8.40-8.38(d,J=7.2Hz,1H),8.28-8.26 (d,J=7.2Hz,1H),7.94-7.88(m,2H),6.87-6.68(m,1H),6.27(s,1H),3.82(br s,1H),1.98-1.68(m,6H),1.52-1.45(m,1H),1.41-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 97.41%. Chiral purity: 98.5% ee.

[0513] INSCoV-517C(4): 1 H NMR(400MHz,DMSO d6): δ9.24-8.87(m,1H),8.44(s,2H),8.41-8.39(d,J=7.2Hz,1H),8.28-8.26 (d,J=7.2Hz,1H),7.95-7.88(m,2H),6.87-6.68(m,1H),6.24(s,1H),3.81(br s,1H),1.98-1.68(m,6H),1.52-1.45(m,1H),1.41-1.23(m,1H). LCMS=[M+H] + :(550.18), purity = 98.90%. Chiral purity: 99.82% ee.

[0514] Example 4. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-N-(4-(oxazole- Synthesis of 5-(5-amino)phenyl)acetamide (INSCoV-501A)

[0515] Option 4

[0516]

[0517] At 25 ° C, 4- (oxazol-5-yl) aniline (148 mg, 0.925 mmol) and pyrimidine-5-carboxaldehyde (100 mg, 0.925 mmol) were added to a solution of 2-chloroacetic acid (87 mg, 0.925 mmol) and isocyanatocyclohexane (101 mg, 0.925 mmol) in 2,2,2-trifluoroethanol (7 mL) at 25 ° C. The mixture was stirred for 1 h at 25 ° C. LC-MS showed that 4- (oxazol-5-yl) aniline was completely consumed, and a main peak with the required mass was detected. The solvent was evaporated under reduced pressure to produce a residue. The crude product was ground with MeOH (15 mL) and washed with MeOH (3 mL × 3). The filter cake was concentrated in vacuo. The residue was diluted with water (10 mL) and lyophilized to produce the product. INSCoV-501A (264.74 mg, 574.76 μmol, 62.13% yield) was obtained as a white solid.

[0518] 1 H NMR (400MHz, DMSO-d6) δ = 8.95 (s, 1H), 8.51-8.40 (m, 3H), 8.20 (d, J = 7.6Hz, 1H), 7.72 (s, 1H), 7.67-7.61 (m, 2H ),7.49-7.43(m,1H),6.10(s,1H),4.15-3.94(m,2H),3.68-3.52(m,1H),1.81-1.46(m,5H),1.35-0.94(m,5H). LCMS:m / z 454.3[M+H] + , purity = 98.5%

[0519] Example 5. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)- Synthesis of N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501I)

[0520] 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-N-(4-(oxazol-5-yl)phenyl)acetamide was synthesized according to the procedure used to prepare INSCoV-501A (Example 4). The crude product was triturated with MTBE (20 mL x 2) and filtered. It was then triturated with MeOH (6 mL) and filtered. INSCoV-501I (214.09 mg, 426.40 μmol, 46.09% yield) was obtained as an off-white solid.

[0521] 1H NMR: (400MHz, DMSO-d6) δ = 8.96 (s, 1H), 8.49 (s, 2H), 8.45 (s, 1H), 8.31 (d, J = 7.6Hz, 1H), 7.71 (s, 1H), 7.65-7.63 (m, 2H), 7.43 (br s,1H),6.07(s,1H),4.11-3.97(m,2H),3.90-3.74(m,1H),2.05-1.71(m,6H),1.59-1.30(m,2H). LCMS:m / z 490.3[M+H] + , purity = 98.9%.

[0522] Example 6. Synthesis of INSCoV-600J, INSCoV-600J(1) and INSCoV-600J(2)

[0523] Option 5

[0524]

[0525] Step 1: 2-chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-N-(4-(isoxazol-5-yl)phenyl)acetamide was synthesized according to the procedure used to prepare INSCoV-501A (Example 4). The crude product was triturated with MTBE (20 mL x 2) and filtered. INSCoV-600J (165.79 mg, 338.42 μmol, 36.58% yield) was obtained as a yellow solid.

[0526] 1 H NMR: (400MHz, DMSO-d6) δ = 8.97 (s, 1H), 8.66 (d, J = 2.0Hz, 1H), 8.50 (s, 2H), 8.34 (d, J = 7.6Hz, 1H), 7.82-7.80 (m, 2H), 7.50-7.49 (m, 2H) ),7.06(d,J=1.9Hz,1H),6.09(s,1H),4.16-4.00(m,2H),3.92-3.75(m,1H),2.06-1.72(m,6H),1.61-1.45(m,1H),1.43-1.29(m,1H). LCMS:m / z 490.2[M+H] + , purity = 100%.

[0527] Step 2: Chiral SFC purification of INSCoV-600J: INSCoV-600J (1) and INSCoV-600J (2). INSCoV-600J (100 mg, 204.12 μmol, 1 eq) was separated by chiral SFC (column: Daicel ChiralPak IG (250*30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 40%-40%, 6.2; 60 min) and concentrated in vacuo. The first peak INSCoV-600J (1) (28.97 mg, 59.13 μmol, 28.97% yield) was obtained as a yellow solid. The second peak INSCoV-600J (2) (22.10 mg, 45.11 μmol, 22.10% yield) was obtained as a yellow solid.

[0528] INSCoV-600J(1): 1 H NMR: (400MHz, DMSO-d6) δ = 8.97 (s, 1H), 8.66 (d, J = 2.0Hz, 1H), 8.50 (s, 2H), 8.33 (d, J = 7.4Hz, 1H), 7.82-7.79 (m, 2H), 7.50 (br s,2H),7.05(d,J=1.8Hz,1H),6.09(s,1H),4.15-3.99(m,2H),3.89-3.78(m,1H),2.04-1.73(m,6H),1.59-1.47(m,1H),1.42-1.30(m,1H). LCMS:m / z 490.3[M+H] + , purity = 96.3%. Chiral purity: 98.5% ee.

[0529] INSCoV-600J(2): 1 H NMR: (400MHz, DMSO-d6) δ = 8.97 (s, 1H), 8.66 (d, J = 2.0Hz, 1H), 8.50 (s, 2H), 8.33-8.31 (m, 1H), 7.82-7.79 (m, 2H), 7 .61-7.35(m,2H),7.05(d,J=1.8Hz,1H),6.09(s,1H),4.13-3.97(m,2H),3.91-3.77(m,1H),2.02-1.72(m,6H),1.59 -1.47(m,1H),1.43-1.32(m,1H). LCMS:m / z 490.3[M+H] + , purity = 99.6%. Chiral purity: 99.0% ee.

[0530] Example 7. Synthesis of INSCoV-600K, INSCoV-600K(1) and INSCoV-600K(2)

[0531] Option 6

[0532]

[0533] Step 1: To a solution of 4-iodoaniline (216.16 mg, 986.95 μmol, 1 eq) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (250 mg, 1.18 mmol, 1.2 eq) in dioxane (7.5 mL) and H2O (2.5 mL) was added Na2CO3 (261.51 mg, 2.47 mmol, 2.5 eq) and Pd(PPh3)4 (57.02 mg, 49.35 μmol, 0.05 eq). The mixture was stirred at 80°C under N2 for 12 h. LCMS showed that one peak with the desired mass was detected. TLC (PE / EA=3 / 1) showed that 4-iodoaniline was consumed and three new spots were formed. The reaction mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL*3). The combined organic layers were dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to yield a residue. The residue was purified by column chromatography (SiO, PE:EA = 20:1 to 2:1). 4-(thiazol-5-yl)aniline (0.15 g, 92.1% purity, 79.5% yield) was obtained as a yellow solid.

[0534] LCMS: m / z 177.2 [M+H] + , purity = 92.1%.

[0535] Step 2: To a solution of 2-chloroacetic acid (188.82 mg, 2.00 mmol, 224.79 μL, 1.2 eq) and pyrimidine-5-carbaldehyde (0.18 g, 1.67 mmol, 1 eq) in CF3CH2OH (10 mL) was added 1,1-difluoro-4-isocyanatocyclohexane (241.70 mg, 1.67 mmol, 1 eq) and 4-(thiazol-5-yl)aniline (293.46 mg, 1.67 mmol, 1 eq). The reaction mixture was stirred at 25°C for 1 h. LCMS showed complete consumption of 4-(thiazol-5-yl)aniline, and a new peak with the desired mass was detected. The reaction mixture was concentrated in vacuo. The reaction mixture was triturated with MTBE (20 mL) and washed with MTBE (10 mL*3). The filter cake was concentrated in vacuo. The residue was diluted with MTBE (20 mL) and washed with MTBE (10 mL*3). The filter cake was concentrated in vacuo. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-N-(4-(thiazol-5-yl)phenyl)acetamide INSCoV-600K was obtained as a yellow solid (177.26 mg, 332.75 μmol, 19.98% yield).

[0536] 1 H NMR (400MHz, DMSO-d6) δ = 9.09 (s, 1H), 8.97 (s, 1H), 8.50 (s, 2H), 8.37-8.28 (m, 2H), 7.64 (d, J = 8.8Hz, 2H), 7 .51-7.09(m,2H),6.07(s,1H),4.12-3.97(m,2H),3.90-3.78(m,1H),2.04-1.70(m,6H),1.58-1.25(m,2H). LCMS:m / z 506.3[M+H] + , purity = 93.0%.

[0537] Step 3: Chiral SFC purification of INSCoV-600K: INSCoV-600K (1) and INSCoV-600K (2). INSCoV-600K (49 mg) was purified by SFC separation (column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 50%-50%, 4 min; 20 min) and vacuum concentrated (<35 ° C). The first peak INSCoV-600K (1) (13 mg, 24.49 μmol, 25% yield, 95.32% purity) was obtained as a yellow solid. The second peak INSCoV-600K (2) (10 mg, 19.11 μmol, 19.7% yield, 96.71% purity) was obtained as a yellow solid.

[0538] INSCoV-600K(1): 1 H NMR (400MHz, DMSO-d6) δ = 9.19-8.84 (m, 2H), 8.51 (s, 2H), 8.33 (s, 2H), 7.64 (d, J = 6.4Hz, 2H), 7.40 ( s,2H),6.08(s,1H),4.09-4.01(m,2H),3.83(d,J=1.6Hz,1H),2.05-1.71(m,6H),1.61-1.29(m,2H). LCMS:m / z 506.3[M+H] + , purity = 99.1%. Chiral purity: 100% ee.

[0539] INSCoV-600K(2): 1 H NMR (400MHz, DMSO-d6) δ = 9.18-8.87 (m, 2H), 8.51 (s, 2H), 8.40-8.24 (m, 2H), 7.63 (d, J = 8.4Hz, 2H), 7.40 (s, 2H) ),6.08(s,1H),4.20-3.96(m,2H),3.92-3.72(m,1H),2.05-1.68(m,6H),1.62-1.47(m,1H),1.43-1.30(m,1H). LCMS:m / z506.2[M+H] + , purity = 100%. Chiral purity: 100% ee.

[0540] Example 8. Synthesis of INSCoV-601G, INSCoV-601G(1) and INSCoV-601G(2)

[0541] Option 7

[0542]

[0543] Step 1: To a solution of 4-(thiazol-5-yl)aniline (150 mg, 851.12 μmol, 1 eq) and 4,4-difluorocyclohexane-1-carbonitrile (123.54 mg, 851.12 μmol, 1 eq) in CF3CH2OH (4 mL) was added 2-chloroacetic acid (80.43 mg, 851.12 μmol, 95.75 μL, 1 eq) and pyrazine-2-carboxaldehyde (92.00 mg, 851.12 μmol, 1 eq). The reaction mixture was stirred at 25 ° C for 1 h. LCMS showed that the reactants were consumed and a peak with the desired mass was detected. The reaction was concentrated in vacuo. The crude product was triturated with MTBE (20 mL) and filtered. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrazin-2-yl)ethyl)-N-(4-(thiazol-5-yl)phenyl)acetamide INSCoV-601G was obtained as a yellow solid (303.51 mg, 584.83 μmol, 68.71% yield).

[0544] 1 H NMR (400MHz, DMSO-d6): δ = 9.09 (d, J = 0.6Hz, 1H), 8.52 (d, J = 1.2Hz, 1H), 8.49-8.48 (m, 1H), 8.44 (d, J = 2.4Hz, 1H), 8.34-8.26(m,2H),7.60-7.58(m,2H),7.55-7.30(m,2H),6.23(s,1H),4.21-3.98(m,2H),3.88-3.68(m,1H),2.01 -1.82(m,4H),1.80-1.70(m,2H),1.53-1.37(m,2H). LCMS:m / z 506.0[M+H] + , purity = 100%.

[0545] Step 2: Chiral SFC purification of INSCoV-601G: INSCoV-601G (1) and INSCoV-601G (2). INSCoV-601G (100 mg, 197.64 μmol, 1 eq) was separated by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm * 30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 55%-55%, 4.4 min; 45 min) and concentrated in vacuo. The first peak INSCoV-601G (1) (34.34 mg, 62.33 μmol, 31.54% yield, 91.837% purity) was obtained as a brown solid. The second peak INSCoV-601G (2) (28.63 mg, 52.47 μmol, 26.55% yield, 92.721% purity) was obtained as a brown solid.

[0546] INSCoV-601G(1): 1 H NMR: (400MHz, DMSO-d6) δ = 9.08 (d, J = 0.6Hz, 1H), 8.55-8.47 (m, 2H), 8.43 (d, J = 2.6Hz, 1H), 8.34-8.24 (m, 2H), 7.60-7.58 (m, 2H) ,7.53-7.28(m,2H),6.23(s,1H),4.19-3.98(m,2H),3.85-3.74(m,1H),2.02-1.83(m,4H),1.79-1.70(m,2H),1.52-1.35(m,2H). LCMS:m / z 506.3[M+H] + , purity = 100%. Chiral purity: 100% ee.

[0547] INSCoV-601G(2): 1 H NMR: (400MHz, DMSO-d6) δ = 9.08 (s, 1H), 8.52 (d, J = 1.2Hz, 1H), 8.49-8.48 (m, 1H), 8.43 (d, J = 2.4Hz, 1H), 8.33-8.27 (m, 2H), 7.60-7.58 ( m,2H),7.52-7.33(m,2H),6.23(s,1H),4.19-4.00(m,2H),3.88-3.73(m,1H),1.99-1.81(m,4H),1.80-1.70(m,2H),1.52-1.37(m,2H). LCMS:m / z 506.2[M+H] + , purity = 100%. Chiral purity: 86.5% ee.

[0548] Example 9. Synthesis of INSCoV-601H

[0549] Option 8

[0550]

[0551] To a solution of 4-(isoxazole-5-yl)aniline (148.17 mg, 925.09 μmol, 1 eq) and 4,4-difluorocyclohexane-1-carbonitrile (134.28 mg, 925.09 μmol, 1 eq) in CF3CH2OH (4 mL) was added 2-chloroacetic acid (87.42 mg, 925.09 μmol, 104.07 μL, 1 eq) and pyrazine-2-carboxaldehyde (100 mg, 925.09 μmol, 1 eq). The reaction mixture was stirred for 1 h at 25 ° C. LCMS showed that the reactants were consumed and a peak with the desired mass was detected. The reaction was concentrated in vacuo. The crude product was ground with MTBE (20 mL) and filtered. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrazin-2-yl)ethyl)-N-(4-(isoxazol-5-yl)phenyl)acetamide INSCoV-601H was obtained as an off-white solid (398.64 mg, 797.28 μmol, 86.18% yield).

[0552] 1 H NMR (400MHz, DMSO-d6): δ = 8.65 (d, J = 2.0Hz, 1H), 8.54 (d, J = 0.8Hz, 1H), 8.50-8.4 6(m,1H),8.43(d,J=2.4Hz,1H),8.31(d,J=7.6Hz,1H),7.78-7.76(m,2H),7.54(br s,2H),7.04(d,J=2.0Hz,1H),6.25(s,1H),4.23-4.01(m,2H),3.85-3.72(m,1H),2.02-1.81(m,4H),1.80-1.70(m,2H),1.52-1.36(m,2H). LCMS: m / z 490.3 [M+H] + , purity = 98.23%.

[0553] Example 10. Synthesis of INSCoV-601I, INSCoV-601I(1) and INSCoV-601I(2)

[0554] Option 9

[0555]

[0556] Step 1: To a solution of 4-iodoaniline (432.33 mg, 1.97 mmol, 1 eq) and 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isothiazole (0.5 g, 2.37 mmol, 1.2 eq) in dioxane (24 mL) and H2O (8 mL) was added Pd(PPh3)4 (228.09 mg, 197.39 μmol, 0.1 eq) and Na2CO3 (523.03 mg, 4.93 mmol, 2.5 eq). The mixture was stirred at 80°C under N2 for 12 h. LCMS showed that 4-iodoaniline (R t =0.866 min) remained, and a peak with the desired mass was detected (R t =0.809 min). TLC (PE:EA=5:1) showed that 4-iodoaniline (R f =0.6) remain and two spots are formed (R f =0.9, R f =0.4). The reaction mixture was diluted with water (10 mL) and extracted with EA (20 ml x 3). The combined organic phases were concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 20g The product was purified by silica flash column (eluent: 0-50% ethyl acetate / petroleum ether at 60 mL / min). The combined organic phases were concentrated in vacuo to obtain 4-(isothiazol-5-yl)aniline (0.1 g, 567.41 μmol, 28.75% yield) as a white solid.

[0557] 1 H NMR (400MHz, DMSO-d6): δ = 8.44 (d, J = 1.6 Hz, 1H), 7.49 (d, J = 2.0 Hz, 1H), 7.43-7.36 (m, 2H), 6.67-6.55 (m, 2H), 5.63 (s, 2H).

[0558] Step 2: Compounds were synthesized according to the procedure for preparing INSCoV-601H (Example 9). MTBE (20 mL) was added to the reaction mixture, filtered and washed with MTBE (10 mL × 3) to give a crude product. The residue was ground with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. 2-chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrazin-2-yl)ethyl)-N-(4-(isothiazol-5-yl)phenyl)acetamide INSCoV-601I (166.44 mg, 322.55 μmol, 58.11% yield) was obtained as a white solid.

[0559] 1 H NMR (400MHz, DMSO-d6): δ = 8.59 (d, J = 1.8Hz, 1H), 8.54 (d, J = 1.2Hz, 1H), 8.52-8.46 (m, 1H), 8.44 (d, J = 2.4Hz, 1H), 8.29 (d, J = 7.6Hz, 1H), 7.78 ( d,J=1.6Hz,1H),7.66(d,J=8.8Hz,2H),7.50(s,2H),6.24(s,1H),4.22 -4.01(m,2H),3.87-3.75(m,1H),2.06-1.68(m,6H),1.54-1.34(m,2H). LCMS:m / z 506.2[M+H] + , purity = 100%.

[0560] Step 3: Chiral SFC purification of INSCoV-601I: INSCoV-601I (1) and INSCoV-601I (2). 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrazin-2-yl)ethyl)-N-(4-(isothiazol-5-yl)phenyl)acetamide INSCoV-601I (0.1 g, 197.64 μmol) was separated by chiral SFC (column: DAICEL CHIRALP AKAD (250 mm × 30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 45%-45%, 4 min; 20 min) to produce the first peak INSCoV-601I (1) (23.1 mg, 44.66 μmol, 22.59% yield, 97.81% purity) as a yellow solid. The second peak INSCoV-601I (2) was obtained as a yellow solid (9.94 mg, 19.52 μmol, 9.87% yield, 99.346% purity).

[0561] INSCoV-601I(1): 1H NMR (400MHz, DMSO-d6): δ=8.61-8.57(m,1H),8.54(d,J=1.2Hz,1H),8.52-8.46(m,1H),8.44(d,J=2.4Hz,1H),8.29(d,J=7.6Hz,1H),7.78(d,J= 1.6Hz,1H),7.66(d,J=8.8Hz,2H),7.61-7.39(m,2H),6.24(s,1H),4.23 -4.01(m,2H),3.85-3.73(m,1H),1.98-1.71(m,6H),1.54-1.35(m,2H). LCMS:m / z 506.3[M+H] + , purity = 100%. Chiral purity: 97.2% ee.

[0562] INSCoV-601I(2): 1 H NMR (400MHz, DMSO-d6) δ=8.59(d,J=1.8Hz,1H),8.54(d,J=1.2Hz,1H),8.52-8.46(m,1H),8.44(d,J=2.4Hz,1H),8.29(d,J=7.6Hz,1H),7.78(d,J =1.6Hz,1H),7.66(d,J=8.8Hz,2H),7.58-7.42(m,2H),6.24(s,1H),4.2 2-4.02(m,2H),3.88-3.75(m,1H),2.05-1.71(m,6H),1.56-1.34(m,2H). LCMS:m / z 506.2[M+H] + , purity = 100%. Chiral purity: 91.76% ee.

[0563] Example 11. Synthesis of INSCoV-601K, INSCoV-601K(1) and INSCoV-601K(2)

[0564] Plan 10

[0565]

[0566] Step 1: Compound is synthesized according to the procedure for preparing INSCoV-601H (Example 9). The mixture is concentrated in vacuo. The crude product is dissolved in MTBE (10 mL), stirred for a moment, and the filter cake is concentrated in vacuo. It is then dissolved in EtOAc (10 mL), stirred for a moment, and the filter cake is concentrated in vacuo. 2- chloro-N- (2- ((4,4- difluorocyclohexyl) amino) -2- oxo -1- (pyrimidin-5-yl) ethyl) -N- (4- (thiazol-5-yl) phenyl) acetamide INSCoV-601K (100 mg, 181.83 μmol, 19.66% yield) as a yellow solid was obtained.

[0567] 1 H NMR (400MHz, DMSO-d6): δ = 8.98 (s, 1H), 8.60 (d, J = 1.8Hz, 1H), 8.51 (s, 2H), 8.33 (d, J = 7.6Hz, 1H), 7.81 (d, J = 1.7Hz, 1H), 7.71 (br d,J=8.8Hz,2H),7.55-7.30(m,2H),6.09(s,1H),4.14-4.09(m,2H),3.93-3.76(m,1H),2.07-1.72(m,6H),1.63-1.21(m,2H). LCMS:m / z506.1[M+H] + , purity = 92.0%.

[0568] Step 2: Chiral SFC purification of INSCoV-601K: INSCoV-601K (1) and INSCoV-601K (2). INSCoV-601K (102 mg) was separated by chiral SFC (column: Daicel ChiralPak IG (250*30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 60%-60%, 4.7; 35 min). The first peak INSCoV-601K (1) (9.97 mg, 18.94 μmol, 9.4% yield, 96.138% purity) was obtained as a yellow solid. The second peak INSCoV-601K (2) (54.54 mg, 104.52 μmol, 51.8% yield, 96.965% purity) was obtained as a yellow solid.

[0569] INSCoV-601K(1): 1H NMR (400MHz, DMSO-d6) δ=8.98(s,1H),8.59(d,J=1.8Hz,1H),8.51(s,2H),8.32(br d,J=7.5Hz,1H),7.80(d,J=1.8Hz,1H),7.70(br d,J=8.7Hz,2H),7.46(br d,J=1.8Hz,2H),6.08(s,1H),4.12-4.03(m,2H),3.91-3.75(m,1H),3.17( d,J=5.3Hz,1H),2.08-1.73(m,7H),1.61-1.48(m,1H),1.45-1.30(m,1H). LCMS: m / z 506.1[M+H] + , purity = 97.97%. Chiral purity: 87.96% ee.

[0570] INSCoV-601K(2): 1 H NMR (400MHz, DMSO-d6) δ=8.97(s,1H),8.58(d,J=1.8Hz,1H),8.50(s,2H),8.31(br d,J=7.5Hz,1H),7.79(d,J=1.8Hz,1H),7.70(br d,J=8.7Hz,2H),7.45(br s,2H),6.08(s,1H),4.14-4.03(m,2H),3.91-3.76(m,1H),3.16(d,J=5.0Hz,1H),2.09-1.71(m,7H),1.61-1.34(m,2H). LCMS:m / z 506.1[M+H] + , purity = 96.12%. Chiral purity: 97.71% ee.

[0571] Example 12. Synthesis of INSCoV-601N, INSCoV-601N(1) and INSCoV-601N(2)

[0572] Plan 11

[0573]

[0574] Step 1: At 0 ° C, under N2, to a solution of 1-(pyrazin-2-yl)ethan-1-one (500 mg, 4.09 mmol, 1 eq) and 4-isothiazol-5-ylaniline (649.40 mg, 3.68 mmol, 2.33 mL, 0.9 eq) in DCM (8 mL) was added TEA (1.24 g, 12.28 mmol, 1.71 mL, 3 eq) and TiCl4 (1 M, 2.05 mL, 0.5 eq). The mixture was stirred for 1 h at 0 ° C, then warmed to 30 ° C and stirred at 30 ° C for 11 h. LCMS showed that 4-isothiazol-5-ylaniline was consumed and 62% of the desired mass was detected. The mixture was diluted with NH4Cl (10 mL), extracted with EtOAc (10 mL×2) and washed with brine (30 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The crude product was purified by Al 2 O 3 chromatography, eluting with DCM / ethyl acetate = 1:0 to 0:1. N-(4-(isothiazol-5-yl)phenyl)-1-(pyrazin-2-yl)ethane-1-imine (0.9 g, 1.64 mmol, 39.99% yield, 51% purity) was obtained as a yellow solid.

[0575] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 9.50 (br s, 1H), 8.82-8.58 (m, 3H), 7.66 (d, J = 8.4 Hz, 2H), 6.93 (d, J = 8.4 Hz, 2H), 6.72 (br d, J = 8.4 Hz, 1H), 2.39 (s, 3H).

[0576] Step 2: To a solution of N-(4-(isothiazol-5-yl)phenyl)-1-(pyrazin-2-yl)ethan-1-imine (0.9 g, 3.21 mmol, 1 eq) in CF3CH2OH (10 mL) was added 1,1-difluoro-4-isocyanato-cyclohexane (465.97 mg, 3.21 mmol, 1 eq) and 2-chloroacetic acid (0.42 g, 4.44 mmol, 500.00 μL, 1.38 eq) at 0°C, and the mixture was stirred at 0°C for 1 h. LCMS showed that N-(4-(isothiazol-5-yl)phenyl)-1-(pyrazin-2-yl)ethan-1-imine was consumed, and 25% of the desired product was detected. The mixture was diluted with water (10 mL), extracted with EtOAc (10 mL x 2), and washed with brine (20 mL). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (column height: 250 mm, diameter: 100 mm, 100-200 mesh silica gel, petroleum ether / ethyl acetate=10 / 1, 1 / 1). 2-(2-chloro-N-(4-(isothiazol-5-yl)phenyl)acetamido)-N-(4,4-difluorocyclohexyl)-2-(pyrazine-2-yl)propionamide INSCoV-601N (283.73 mg, 529.92 μmol, 16.51% yield, 97.119% purity) as a yellow solid was obtained.

[0577] 1 H NMR (400MHz, DMSO-d6): δ = 9.06 (d, J = 1.2Hz, 1H), 8.69-8.53 (m, 3H), 8.25 (d, J = 8.0Hz, 1H), 7.97-7.79 (m, 4H), 7.58 (br d,J=8.0Hz,1H),4.13-3.94(m,2H),3.87(br d,J=7.2Hz,1H),2.11-1.50(m,8H),1.42(s,3H). LCMS:m / z 520.1[M+H] + , purity = 98.61%.

[0578] Step 3: Chiral SFC purification of INSCoV-601N: INSCoV-601N (1) and INSCoV-601N (2). INSCoV-601N (200 mg) was purified by SFC (column: DAICEL CHIRALCEL OD (250 mm × 30 mm, 10 μm); mobile phase: [Neu-IPA]; B%: 40%-40%, 4.0 min; 25 min) to produce two peaks. The first peak INSCoV-601N (1) (52.24 mg, 91.42 μmol, 23.77% yield, 91% purity) was obtained as an off-white solid. The second peak INSCoV-601N (2) (51.09 mg, 92.36 μmol, 24.01% yield, 94% purity) was obtained as an off-white solid.

[0579] INSCoV-601N(1): 1 H NMR (400MHz, DMSO-d6): δ = 9.06 (d, J = 1.2Hz, 1H), 8.68-8.59 (m, 2H), 8.57 (d, J = 2.4Hz, 1H), 8.25(d,J=8.0Hz,1H),7.96-7.80(m,4H),7.58(d,J=8.0Hz,1H),4.12-3.94(m,2H),3.87(br d,J=7.6Hz,1H),2.14-1.51(m,8H),1.42(s,3H). LCMS:m / z520.1[M+H] + , purity = 94.81%. Chiral purity: 100% ee.

[0580] INSCoV-601N(2): 1 H NMR (400MHz, DMSO-d6): δ = 9.06 (d, J = 1.2Hz, 1H), 8.68-8.60 (m, 2H), 8.57 (d, J = 2.5Hz, 1H), 8.25 (d, J = 8.0Hz, 1H), 7.94-7.80 (m, 4H), 7.58 (br d,J=8.0Hz,1H),4.09-3.95(m,2H),3.88(br d,J=7.2Hz,1H),2.11-1.52(m,8H),1.42(s,3H). LCMS:m / z520.1[M+H] + , purity = 100%. Chiral purity: 100% ee.

[0581] Example 13. Synthesis of INSCoV-601P, INSCoV-601P(1A) and INSCoV-601P(1B)

[0582] Plan 12

[0583]

[0584] Step 1: To a solution of 4-(isoxazol-5-yl)aniline (2.6 g, 16.23 mmol, 1 eq) and 1-(pyrimidin-5-yl)ethan-1-one (2.38 g, 19.48 mmol, 1.2 eq) in DCM (26 mL) was added TEA (4.93 g, 48.70 mmol, 6.78 mL, 3 eq) and TiCl4 (1 M, 8.12 mL, 0.5 eq) at 0°C. The reaction mixture was stirred at 25°C under N2 for 12 h. LCMS showed complete consumption of the reactants and a peak of the desired mass was detected (R t =0.884min). The reaction mixture was filtered through a celite pad and washed with DCM (40mL*2). The filtrate was concentrated in vacuo. The residue was used in the next step without further purification. N-(4-(isothiazol-5-yl)phenyl)-1-(pyrazin-2-yl)ethane-1-imine (4.5 g, crude product) was obtained as a yellow solid.

[0585] LCMS: m / z 265.2 [M+H] + , purity = 63.77%.

[0586] Step 2: To a solution of 2-chloroacetic acid (1.93 g, 20.43 mmol, 2.30 mL, 1.2 eq) and N-(4-(isothiazol-5-yl)phenyl)-1-(pyrazin-2-yl)ethane-1-imine (4.5 g, 17.03 mmol, 1 eq) in CF3CH2OH (15 mL) was added 1,1-difluoro-4-isocyanatocyclohexane (2.47 g, 17.03 mmol, 1 eq). The reaction mixture was stirred at 25°C for 12 h. LCMS showed that one peak with the desired mass (R t=0.929 min). The reaction was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex luna C18 (250*70 mm, 15 μm); mobile phase: [water (0.05% HCl)-ACN]; B%: 35 ACN%-65 ACN%, 22 min), diluted with water (800 mL) and dried by lyophilization to give a crude product. The residue was purified by normal phase HPLC (column: Welch Ultimate XB-SiOH 250*50*10 μm; mobile phase: [hexane-EtOH]; B%: 1%-40%, 20 min) and concentrated in vacuo. 2-(2-Chloro-N-(4-(isoxazol-5-yl)phenyl)acetamido)-N-(4,4-difluorocyclohexyl)-2-(pyrimidin-5-yl)propanamide (INSCoV-601P) was obtained as a yellow solid (0.2 g, 360.54 μmol, 2.12% yield).

[0587] 1 H NMR (400MHz, DMSO-d6): δ = 9.04 (s, 1H), 8.89-8.83 (m, 2H), 8.69 (d, J = 2.0Hz, 1H), 8.03-7.79 (m, 3H), 7.66-7.59 (m,1H),7.37(d,J=7.6Hz,1H),7.13(d,J=2.0Hz,1H),4.07-3.91(m,2H),3.90-3.81(m,1H),2.07-1.56(m,11H). LCMS:m / z 504.2[M+H] + , purity = 97.55%.

[0588] Step 3: Chiral SFC purification of INSCoV-601P: INSCoV-601P (1A) and INSCoV-601P (1B). INSCoV-601P (0.2 g, 396.88 μmol, 1 eq) was separated by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm*30 mm, 10 μm); mobile phase: [Neu-EtOH]; B%: 45%-45%, 5.2; 40 min) and concentrated in vacuo. Then, the two peaks were separated by chiral SFC (column: DAICEL CHIRALPAK IG (250 mm * 50 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 40%-40%, 4.3 min; 25 min) and chiral SFC (column: Daicel Chiral Pak IG (250 * 30 mm, 10 μm); mobile phase: [MeOH-ACN]; B%: 50%-50%, 4.1; 40 min), respectively, and concentrated in vacuo. The first peak INSCoV-601P (1A) (45.6 mg, 85.98 μmol, 22% yield) was obtained as a yellow solid. The second peak INSCoV-601P (1B) (53.71 mg, 97.77 μmol, 25% yield) was obtained as an orange solid.

[0589] INSCoV-601P(1A): 1 H NMR (400MHz, chloroform-d) δ = 9.17 (s, 1H), 8.86 (s, 2H), 8.35 (d, J = 2.0Hz, 1H), 7.92 (s, 2H), 7.57-7.34 (m, 2H), 6.63 (d, J =1.6Hz,1H),6.51(d,J=7.2Hz,1H),4.10-3.94(m,1H),3.88-3.73(m,2H),2.18-1.86(m,9H),1.66-1.57(m,2H). LCMS:m / z504.2[M+H] + , purity = 96.07%. Chiral purity: 100% ee.

[0590] INSCoV-601P(1B): 1H NMR (400MHz, chloroform-d) δ = 9.13 (s, 1H), 8.82 (s, 2H), 8.34 (d, J = 2.0Hz, 1H), 7.90 (s, 2H), 7.56-7.30 (m, 2H), 6.62 (d, J = 2 .0Hz,1H),6.50(d,J=7.6Hz,1H),4.01-3.95(m,1H),3.85-3.73(m,2H),2.23-1.83(m,9H),1.60(d,J=10.0Hz,2H). LCMS:m / z 504.1[M+H] + , purity = 97.39%. Chiral purity: 100% ee.

[0591] Example 14. Synthesis of INSCoV-601Q, INSCoV-601Q(1A) and INSCoV-601Q(1B)

[0592] Plan 13

[0593]

[0594] Step 1: To a solution of (S)-tetrahydrofuran-3-amine (4 g, 32.37 mmol, 1 eq, HCl) in ethyl formate (36.84 g, 497.31 mmol, 40.00 mL, 15.36 eq) was added TEA (9.83 g, 97.10 mmol, 13.52 mL, 3 eq). The mixture was stirred at 80°C for 17 h. TLC (DCM:MeOH=10:1) showed that (S)-tetrahydrofuran-3-amine (R f =0.4) is consumed and a new spot is formed (R f =0.6). The mixture was concentrated in vacuo. The residue was dissolved in DCM (80 mL) and washed with H2O (30 mL*3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The crude product was used directly in the next step without further purification. (S)-N-(tetrahydrofuran-3-yl)formamide (3.8 g, crude) was obtained as a yellow oil.

[0595] 1 H NMR (400 MHz, chloroform-d): δ = 8.06 (s, 1H), 6.85 (br s, 1H), 4.56-4.46 (m, 1H), 3.93-3.83 (m, 1H), 3.81-3.70 (m, 2H), 3.67-3.56 (m, 1H), 2.30-2.15 (m, 1H), 1.87-1.72 (m, 1H).

[0596] Step 2: To a solution of (S)-N-(tetrahydrofuran-3-yl)formamide (4 g, 34.74 mmol, 1 eq) in DCM (40 mL) was added PPh 3 (9.11 g, 34.74 mmol, 1 eq), CCl 4 (5.34 g, 34.74 mmol, 3.34 mL, 1 eq) and TEA (3.87 g, 38.22 mmol, 5.32 mL, 1.1 eq). The mixture was stirred at 45° C. under N 2 for 17 h. TLC (PE:EA=5:1) showed that (S)-N-(tetrahydrofuran-3-yl)formamide was consumed and a new spot (R f =0.5). The mixture was concentrated in vacuo. The residue was dissolved in Et2O (300 mL), stirred for 30 min, filtered, and the filter cake was washed with Et2O (100 mL*3). The combined organic layers were concentrated in vacuo. (S)-3-isocyanatotetrahydrofuran (4 g, crude product) was obtained as a yellow oil.

[0597] 1 H NMR (400MHz, chloroform-d): δ = 4.16 (br d, J = 4.0Hz, 1H), 4.00 (q, J = 7.9Hz, 1H), 3.95-3.84 (m, 2H), 2.24-2.17 (m, 1H), 1.92 (br s, 2H).

[0598] Step 3: To a solution of (S)-3-isocyanatotetrahydrofuran (179.68 mg, 1.85 mmol, 1 eq) and 4-(isoxazol-5-yl)aniline (296.35 mg, 1.85 mmol, 1 eq) in CF3CH2OH (6 mL) was added pyrazine-2-carboxaldehyde (0.2 g, 1.85 mmol, 1 eq) and 2-chloroacetic acid (174.84 mg, 1.85 mmol, 208.14 μL, 1 eq). The mixture was stirred at 30°C for 1 h. LCMS showed that the reactants were consumed and the desired mass was detected. 30 mL of MTBE was added to the mixture, stirred for 30 min, filtered, and the filter cake was concentrated in vacuo. 30 mL of MTBE was added to the mixture, stirred for 30 min, filtered, and the filter cake was concentrated in vacuo. 2-Chloro-N-(4-(isoxazol-5-yl)phenyl)-N-(2-oxo-1-(pyrazin-2-yl)-2-(((S)-tetrahydrofuran-3-yl)amino)ethyl)acetamide (INSCoV-601Q) was obtained as a yellow solid (461.14 mg, 990.78 μmol, 53.55% yield).

[0599] 1H NMR (400MHz, DMSO-d6): δ = 8.65 (d, J = 2.0Hz, 1H), 8.59-8.52 (m, 2H), 8.50-8.46 (m, 1H), 8.42 (d, J = 2.4Hz, 1H), 7.77 (br d,J=7.7Hz,2H),7.57-7.50(m,1H),7.03(d,J=1.8Hz,1H),6.26(d,J=2.1Hz,1H),4.33-4.23(m,1H),4.18 -4.02(m,2H),3.87(dq,J=6.6,9.7Hz,1H),3.77-3.61(m,3H),2.06(td,J=7.5,12.7Hz,1H),1.75-1.59(m,1H). LCMS:m / z 442.2[M+H] + , purity = 85.55%.

[0600] Step 4: Chiral SFC purification of INSCoV-601Q: INSCoV-601Q (1A) and INSCoV-601Q (1B). Compound INSCoV-601Q (103 mg) was purified by SFC (column: Cellucoat 50×4.6 mm ID, 3 μm, mobile phase: phase A is CO2, and phase B is MeOH (0.05% DEA); gradient elution: 5% to 40% MeOH (0.05% DEA) / CO2; flow rate: 3 mL / min; detector: PDA; column temperature: 35°C; back pressure: 100 bar). The solution was concentrated in vacuo. The first peak INSCoV-601Q (1A) (10.30 mg, 22.43 μmol, 9.91% yield, 96.243% purity) was obtained as a yellow solid. The second peak INSCoV-601Q (1B) was obtained as a yellow solid (28.04 mg, 60.92 μmol, 26.92% yield, 96.007% purity).

[0601] INSCoV-601Q(1A): 1H NMR (400MHz, DMSO-d6): δ = 8.65 (d, J = 2.0Hz, 1H), 8.58-8.53 (m, 2H), 8.50-8.46 (m, 1H), 8.42 (d, J = 2.4Hz, 1H), 7.77 (br d,J=8.7Hz,2H),7.65-7.38(m,2H),7.03(d,J=1.8Hz,1H),6.26(s,1H),4.29-4.22(m,1H),4. 18-4.03(m,2H),3.76-3.63(m,3H),3.42-3.38(m,1H),2.08-1.99(m,1H),1.73-1.63(m,1H). LCMS: m / z 442.2 [M+H] + , purity = 96.27%. Chiral purity: 69.84% ee.

[0602] INSCoV-601Q(1B): 1 H NMR (400MHz, DMSO-d6): δ = 8.65 (d, J = 2.0Hz, 1H), 8.59-8.54 (m, 2H), 8.48 (dd, J = 1.5, 2.4Hz, 1H), 8.43 (d, J = 2.6Hz, 1H), 7.78 (br d,J=8.8Hz,2H),7.67-7.41(m,2H),7.04(d,J=1.8Hz,1H),6.27(s,1H),4.33-4.25(m,1H),4.18-4 .03(m,2H),3.76-3.62(m,2H),3.41(dd,J=3.5,9.0Hz,1H),2.08-2.02(m,1H),1.71-1.62(m,1H). LCMS: m / z 442.2 [M+H] + , purity = 98.23%. Chiral purity: 74.77% ee.

[0603] Example 15. INSCoV-614, INSCoV-614(1A), INSCoV-614(1B), INSCoV-614(2A) and Synthesis of INSCoV-614(2B)

[0604] Plan 14

[0605]

[0606] Step 1: To a solution of ethyl 2-chloro-2-fluoroacetate (1 g, 7.12 mmol, 826.45 μL, 1 eq) in THF (4 mL), MeOH (4 mL) and H2O (2 mL) was added NaOH (426.92 mg, 10.67 mmol, 1.5 eq). The reaction mixture was stirred at 25 ° C for 12 h. TLC (PE: EA = 5: 1) showed the formation of a new spot. The reaction mixture was concentrated in vacuo, pH = 2 was adjusted by 1N HCl solution, and extracted with EtOAc (50 mL * 3). The combined organic phases were dried over anhydrous Na2SO4 and concentrated in vacuo. The reaction mixture was used directly in the next step without further purification. 2-chloro-2-fluoroacetic acid (0.6 g, crude product) was obtained as a colorless oil.

[0607] 1 H NMR (400MHz, DMSO-d6): δ = 7.03-6.65 (m, 1H).

[0608] Step 2: compound is synthesized according to the procedure for preparing INSCoV-601H (Example 9). MTBE (20 mL) is added to the reaction mixture and cooled to 0 ° C for 12 h. The mixture is filtered and washed with MTBE (10 mL * 3). The filter cake is concentrated in vacuo. 2- chloro-N- (2- ((4,4- difluorocyclohexyl) amino) -2- oxo -1- (pyrimidin-5-yl) ethyl) -2- fluoro- N- (4- (thiazol-5-yl) phenyl) acetamide INSCoV-614 (0.5 g, 910.33 μmol, 32.75% yield) as an orange solid is obtained.

[0609] 1 H NMR (400MHz, DMSO-d6): δ = 9.11 (d, J = 0.8Hz, 1H), 9.10 (d, J = 0.8Hz, 1H), 8.99 (s, 1H),8.96(s,1H),8.53(s,2H),8.51(s,1H),8.39-8.35(m,1H),8.34(d,J=4.4Hz ,2H),7.64(s,4H),6.66-6.52(m,1H),6.51-6.40(m,1H),6.09(s,1H),6.03(s,1 H),3.89-3.83(m,2H),2.13-1.64(m,12H),1.60-1.46(m,2H),1.42-1.26(m,2H). LCMS: m / z 524.1[M+H] + , purity = 100%.

[0610] Step 3: Chiral SFC purification of INSCoV-614: INSCoV-614 (1A), INSCoV-614 (1B), INSCoV-614 (2A) and INSCoV-614 (2B). Compound INSCoV-614 (0.4 g, 763.42 μmol, 1 eq) was separated by chiral SFC (column: Daicel ChiralPak IG (250*30 mm, 10 μm); mobile phase: [Neu-MeOH]; B%: 40%-40%, 5.6; 90 min) and concentrated in vacuo. The first peak INSCoV-614 (1A) (50.34 mg, 93.52 μmol, 12.25% yield) was obtained as a yellow solid. The second peak INSCoV-614 (1B) (66.09 mg, 119.03 μmol, 15.59% yield) was obtained as a yellow solid. The third peak, INSCoV-614 (2A), was obtained as a yellow solid (33.89 mg, 60.34 μmol, 7.90% yield). The fourth peak, INSCoV-614 (2B), was obtained as a yellow solid (72.98 mg, 133.83 μmol, 17.53% yield).

[0611] INSCoV-614(1A): 1 H NMR (400MHz, DMSO-d6): δ=9.11(s,1H),8.99(s,1H),8.53(s,2H),8.38-8.31(m,2H),7.68-7.62(m,3H),7.49-7.24( m,1H),6.72-6.38(m,1H),6.03(s,1H),3.89-3.76(m,1H),2.01-1.71(m,6H),1.59-1.45(m,1H),1.43-1.28(m,1H). LCMS:m / z524.1[M+H] + , purity = 97.28%. Chiral purity: 100% ee.

[0612] INSCoV-614(1B): 1H NMR (400MHz, DMSO-d6): δ = 9.09 (s, 1H), 8.96 (s, 1H), 8.51 (s, 2H), 8.38 (d, J = 7.6Hz, 1H), 8.33 (s, 1H), 7.74-7.55 (m, 3H), 7.48 -7.11(m,1H),6.59-6.38(m,1H),6.09(s,1H),3.85(d,J=6.4Hz,1H),2.04-1.73(m,6H),1.62-1.44(m,1H),1.42-1.25(m,1H). LCMS:m / z 524.1[M+H] + , purity = 98.36%. Chiral purity: 96.75% ee.

[0613] INSCoV-614(2A): 1 H NMR (400MHz, DMSO-d6): δ = 9.10 (s, 1H), 8.96 (s, 1H), 8.51 (s, 2H), 8.36 (d, J = 7.6Hz, 1H), 8.34 (s, 1H), 7.66-7.60 (m, 3H), 7.4 9-7.22(m,1H),6.58-6.41(m,1H),6.09(s,1H),3.89-3.83(m,1H),2.03-1.71(m,6H),1.60-1.45(m,1H),1.41-1.25(m,1H). LCMS:m / z 524.1[M+H] + , purity = 98.01%. Chiral purity: 100% ee.

[0614] INSCoV-614(2B): 1 H NMR (400MHz, DMSO-d6): δ = 9.12 (s, 1H), 9.00 (s, 1H), 8.55 (s, 2H), 8.45-8.32 (m, 2H), 7.69-7.63 (m, 3H), 7.27-7.06 ( m,1H),6.73-6.41(m,1H),6.04(s,1H),3.93-3.73(m,1H),2.06-1.70(m,6H),1.61-1.45(m,1H),1.43-1.26(m,1H).

[0615] LCMS: m / z 524.1[M+H] + , purity = 98.39%. Chiral purity: 99.26% ee.

[0616] Based on modeling and activity data, INSCoV-614(1B) is expected to have structure.

[0617] Example 16. INSCoV-614A, INSCoV-614A(1A), INSCoV-614A(1B), INSCoV-614A(2A) and synthesis of INSCoV-614A(2B)

[0618] Plan 15

[0619]

[0620] Step 1: To a solution of 2-chloro-2-fluoroacetic acid (421 mg, 3.74 mmol, 1.2 eq) and pyrimidine-5-carboxaldehyde (338 mg, 3.13 mmol, 1.00 eq) in CF3CH2OH (10 mL) was added 4-(isoxazol-5-yl)aniline (500 mg, 3.12 mmol, 1 eq) and 1,1-difluoro-4-isocyanatocyclohexane (453 mg, 3.12 mmol, 1 eq). The mixture was stirred at 25 ° C for 1 h. LCMS showed that the starting material was completely consumed and about 72% of the desired product was detected. The reaction mixture was concentrated under reduced pressure to produce a crude product. The residue was purified by flash silica gel chromatography ( 25g Purification was performed on a silica flash column (eluent: 0-85% ethyl acetate / petroleum ether at 40 mL / min). 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-2-fluoro-N-(4-(isoxazol-5-yl)phenyl)acetamide (INSCoV-614A) (1.05 g, 1.96 mmol, 63% yield) was obtained as a yellow solid.

[0621] 1 H NMR (400MHz, DMSO-d6): δ=9.02-8.92(m,1H),8.72-8.63(m,1H),8.56-8.48(m,2H),8.43-8.32(m,1H) ,7.81(brs,2H),7.69-7.19(m,2H),7.12-7.02(m,1H),6.68-6.43(m,1H),6.14-6.00(m,1H),3.86(br s,1H),2.12-2.00(m,1H),1.96-1.68(m,5H),1.62-1.45(m,1H),1.40-1.25(m,1H). LCMS:m / z 508.2[M+H] + , purity = 100%.

[0622] Step 2: Chiral SFC purification of INSCoV-614A: INSCoV-614A (1A), INSCoV-614A (1B), INSCoV-614A (2A) and INSCoV-614A (2B). Compound INSCoV-614A (1.05 g, 1.96 mmol, 95% purity, 1 eq) was separated by chiral SFC (Phenomenex-cellulose-2 (250 mm * 50 mm, 10 μm); mobile phase: A: supercritical CO2, B: Neu-EtOH; isocratic: A: B = 70:30; flow rate: 200 mL / min) and concentrated in vacuo to provide four fractions. The first peak INSCoV-614A (1B) was obtained as a yellow solid: (112 mg, 216.54 μmol, 98.194% purity, 11% yield). The second peak INSCoV-614A (1A) was obtained as a white solid: (144 mg, 280.18 μmol, 98.82% purity, 14.3% yield). The third peak INSCoV-614A (2B) was obtained as a yellow solid (120 mg, 231.17 μmol, 97.841% purity, 11.8% yield). The fourth peak INSCoV-614A (2A) was obtained as a yellow solid (202 mg, 389.26 μmol, 97.872% purity, 19.9% ​​yield).

[0623] INSCoV-614A(1A): 1 H NMR (400MHz, DMSO-d6): 8.98(s,1H),8.67(d,J=1.6Hz,1H),8.53(s,2H),8.36(d,J=7.6Hz,1H),7.83(d,J=6.8Hz,2H),7.74-7.18(m,2 H),7.08(d,J=1.6Hz,1H),6.69-6.47(m,1H),6.04(s,1H),3.91-3.75(m,1H),2.05-1.71(m,6H),1.60-1.45(m,1H),1.40-1.27(m,1H). LCMS:m / z 508.1[M+H] + , purity = 100%. Chiral purity: 95.48% ee.

[0624] INSCoV-614A(1B): 1H NMR (400MHz, DMSO-d6):8.97(s,1H),8.67(d,J=2.0Hz,1H),8.53(s,2H),8.36(d,J=7.6Hz,1H),7.90-7.75(m,2H),7.74-7.17(m,2H) ,7.07(d,J=1.6Hz,1H),6.69-6.49(m,1H),6.04(s,1H),3.91-3.76(m,1H),2.06-1.68(m,6H),1.59-1.43(m,1H),1.39-1.26(m,1H). LCMS:m / z 508.0[M+H] + , purity = 100%. Chiral purity: 98.72% ee.

[0625] INSCoV-614A(2A): 1 H NMR (400MHz, DMSO-d6): 8.95 (s, 1H), 8.67 (d, J = 2.0Hz, 1H), 8.51 (s, 2H), 8.39 (d, J = 7.6Hz, 1H), 7. 95-7.75(m,2H),7.73-7.14(m,2H),7.06(d,J=1.8Hz,1H),6.67-6.40(m,1H),6.11(s,1H),3.87(br s,1H),2.07-1.71(m,6H),1.60-1.45(m,1H),1.42-1.25(m,1H). LCMS:m / z 508.1[M+H] + , purity = 96.46%. Chiral purity: 95.82% ee.

[0626] INSCoV-614A(2B): 1 H NMR(400MHz, DMSO-d6):8.95(s,1H),8.66(d,J=2.0Hz,1H),8.51(s,2H),8.39(d,J=7.6Hz,1H),7.97-7.75(m,2H),7.74-7.24(m,2H) ,7.06(d,J=1.6Hz,1H),6.63-6.42(m,1H),6.11(s,1H),3.94-3.79(m,1H),2.04-1.72(m,6H),1.60-1.45(m,1H),1.41-1.25(m,1H). LCMS:m / z 508.1[M+H] + , purity = 97.34%. Chiral purity: 92.96% ee.

[0627] Based on modeling, INSCoV-614A(2A) is expected to have structure.

[0628] Example 17. Synthesis of INSCoV-110A, INSCoV-110A(1) and INSCoV-110A(2)

[0629] Plan 16

[0630]

[0631] To a solution of 2-chloroacetic acid (88.22 mg, 933.62 μmol, 105.03 μL, 1 eq) and isocyanocyclohexane (101.92 mg, 933.62 μmol, 116.08 μL, 1 eq) in 75-89-8 (5 mL) was added 4-tert-butylaniline (139.33 mg, 933.62 μmol, 147.44 μL, 1 eq) and pyridine-3-carboxaldehyde (0.1 g, 933.62 μmol, 87.72 μL, 1 eq). The reaction mixture was stirred for 1 h at 15 ° C. LCMS showed that SM was consumed and DP was formed. The solvent was evaporated under reduced pressure. The crude product was refluxed with PE / EA (10 / 1, 50 mL) and filtered to give 2-(4-tert-butyl-N-(2-chloroacetyl)anilino)-N-cyclohexyl-2-(3-pyridyl)acetamide (0.4 g, 904.99 μmol, 96.93% yield) as a white solid.

[0632] Resolution of INSCoV-110A by SFC afforded INSCoV-110A (1) as a yellow solid: (511.15 mg, 1.16 mmol, 34.08% yield), which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0633] LCMS: retention time: 1.537 min, [M+H + ] = 442.2. HPLC: retention time: 2.778 min. SFC: retention time: 1.163 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.30 (dd, J = 1.6, 4.8Hz, 1H), 8.26 (d, J = 1.9Hz, 1H), 8.05 (d, J = 7.6Hz, 1H), 7.29 (td, J = 1.9, 7.9Hz, 1H), 7.21 (br d,J=6.6Hz,2H),7.09(dd,J=4.8,7.8Hz,1H),6.01(s,1H),4.00-3.86(m,2H),3.61-3.50(m,1H),1.78-1.47(m,5H),1.34-0.91(m,16H).

[0634] Resolution of INSCoV-110A by SFC afforded INSCoV-110A (2) as a white solid: (76.78 mg, 173.71 μmol, 19.19% yield, 100% purity), which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0635] LCMS: retention time: 1.509 min, [M+H + ] = 442.2, 1. HPLC: retention time: 2.080 min. SFC: retention time: 2.087 min. 1 H NMR (400MHz, DMSO-d6): δ=8.47-8.36(m,2H),8.16-8.05(m,1H),7.61-7.49(m,1H),7.35-7.28(m,1H),7.28- 7.22(m,2H),7.22-7.08(m,1H),6.07-6.04(m,1H),4.02-3.90(m,2H),1.75-1.48(m,5H),1.28-1.00(m,17H).

[0636] Example 18. Synthesis of INSCoV-110B, INSCoV-110B(1) and INSCoV-110B(2)

[0637] Plan 17

[0638]

[0639] To a solution of acrylic acid (67.28 mg, 933.62 μmol, 64.08 μL, 1eq) and isocyanatocyclohexane (101.92 mg, 933.62 μmol, 116.08 μL, 1eq) in 75-89-8 (5 mL) was added 4-tert-butylaniline (139.33 mg, 933.62 μmol, 147.44 μL, 1eq) and pyridine-3-carboxaldehyde (0.1 g, 933.62 μmol, 87.72 μL, 1eq). The reaction mixture was stirred for 1 h at 15 ° C. LCMS showed that SM was consumed and DP was formed. The solvent was evaporated under reduced pressure. The crude product was purified by preparative HPLC (TFA) to produce INSCoV-110B (0.23 g, 548.20 μmol, 58.72% yield) as a white solid.

[0640] LCMS: retention time: 0.902 min, [M+H + ] = 420.0. HPLC: retention time: 2.779 min, 1. 1H NMR (400MHz, DMSO-d6): δ=8.47-8.25(m,2H),8.08(d,J=7.7Hz,1H),7.39(br d,J=7.9Hz,1H),7.23(br d,J=8.3Hz,2H),7.17(dd,J=4.8,7.9Hz,1H),7.09(br d,J=2.2Hz,1H),6.26-6.10(m,2H),5.86(dd,J=10.3,16.8Hz,1H),5.66 -5.48(m,1H),3.62-3.52(m,1H),1.81-1.45(m,5H),1.32-0.92(m,15H).

[0641] Resolution of INSCoV-110B by SFC afforded INSCoV-110B (1) as a white solid: (56.13 mg, 133.78 μmol, 24.40% yield), which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0642] LCMS: retention time: 0.909 min, [M+H + ] = 420.0. HPLC: retention time: 3.011 min. SFC: retention time: 0.946 min. 1 H NMR (400MHz, DMSO-d6): δ=8.36-8.24(m,2H),8.06(br d,J=7.7Hz,1H),7.33(br d,J=6.5Hz,1H),7.21(br d,J=7.3Hz,2H),7.15-6.97(m,2H),6.22-6.09(m,2H),5.92-5.78(m,1H),5.56(br d,J=10.6Hz,1H),3.62-3.51(m,1H),1.77-1.47(m,5H),1.32-0.95(m,15H).

[0643] Resolution of INSCoV-110B by SFC afforded INSCoV-110B (2) as a white solid: (59.71 mg, 142.32 μmol, 25.96% yield), which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0644] LCMS: retention time: 0.905 min, [M+H + ] = 442.2. HPLC: retention time: 3.029 min. SFC: retention time: 1.905 min, 1HNMR: (400MHz, DMSO-d6): δ=8.40-8.28(m,2H),8.07(br d,J=7.7Hz,1H),7.39(br d,J=7.9Hz,1H),7.22(br d,J=8.1Hz,2H),7.17(br dd,J=5.0,7.6Hz,1H),7.13-7.01(m,1H),6.22-6.10(m,2H),5.92-5.79(m,1 H),5.62-5.51(m,1H),3.56(brs,1H),1.73-1.50(m,5H),1.32-0.97(m,16H).

[0645] Example 19. Synthesis of INSCoV-110, INSCoV-110-1 and INSCoV-110-2

[0646] Plan 18

[0647]

[0648] At 0 ° C, to a solution of compound 4 (200 mg, 547.18 μ mol, 1 eq) and TEA (166.11 mg, 1.64 mmol, 228.48 μ L, 3 eq) in DCM (8 mL) was added ethylene sulfonyl chloride (138.50 mg, 1.09 mmol, 2.0 eq) dropwise and the reaction was stirred at 20 ° C for 2 h. LCMS showed that compound 4 was consumed and the desired mass was detected as the main peak. The reaction mixture was concentrated. The residue was purified by flash silica gel chromatography ( 25g Purification was performed by silica flash column, 0-45% ethyl acetate / petroleum ether gradient eluent at 50 mL / min. Compound 6 (INSCoV-110) (110 mg, 229.60 μmol, 41.96% yield, 95.1% purity) was obtained as an off-white solid, which was confirmed by LCMS.

[0649] LCMS: retention time: 0.929 min, (M+H)=456.1

[0650] Compound 6 (INSCoV-110) (110 mg, 241.43 μmol, 1 eq) was separated by preparative SFC (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 um); mobile phase: [0.1% NH3H2O ​​MEOH]; B%: 30%-30%, 4.7 min; 35 min).

[0651] INSCoV-110-1 was obtained as a white solid: (15 mg, 32.04 μmol, 13.27% yield, 97.304% purity), which was confirmed by HNMR, LCMS, SFC and HPLC.

[0652] LCMS: retention time: 1.085 min, (M+H) = 456.5. HPLC: retention time: 2.738 min. 1 H NMR (400MHz, chloroform-d): δ = 8.49 (dd, J = 1.7, 4.8Hz, 1H), 8.40 (d, J = 2.2Hz, 1H), 7.39-7.32 (m ,1H),7.24-7.16(m,2H),7.10(dd,J=4.9,7.9Hz,1H),7.07-6.98(m,2H),6.82(dd,J=9.9 ,16.6Hz,1H),6.11(d,J=16.6Hz,1H),5.93(d,J=9.9Hz,1H),5.89-5.80(m,2H),3.92-3. 72(m,1H),2.06-1.93(m,1H),1.93-1.82(m,1H),1.81-1.68(m,2H),1.48-0.97(m,16H).

[0653] INSCoV-110-2 was obtained as a white solid: (20 mg, 42.90 μmol, 17.77% yield, 97.721% purity), which was confirmed by HNMR, LCMS, SFC and HPLC.

[0654] LCMS: retention time: 1.085 min, (M+H) = 456.5. HPLC: retention time: 2.742 min. 1 H NMR (400MHz, chloroform-d): δ=8.50(dd,J=1.6,4.8Hz,1H),8.40(d,J=2.2Hz,1H),7.35(td,J=1.9,8.0Hz,1H),7.24-7.17(m,2H),7.10(dd,J=4.8,7.9H z,1H),7.06-7.00(m,2H),6.82(dd,J=9.9,16.5Hz,1H),6.11(d,J=16.6Hz,1H),5.93(d,J=9.9Hz,1H),5.84(s,1H),3.91-3.75(m,1H),1.98(br d,J=8.6Hz,1H),1.88(br d,J=11.0Hz,1H),1.79-1.69(m,2H),1.47-1.02(m,17H).

[0655] Example 20. 2-Chloro-N-(2-((1,1-dioxotetrahydro-2H-thiopyran-4-yl)amino)-2-oxo-1-(pyrimidin ... Synthesis of N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501B)

[0656] INSCoV-501B was obtained according to the general procedure for the INSCoV series and purified by the following purification B: the crude product was triturated with MTBE (20 mL × 2) and filtered. It was then triturated with MeOH (6 mL) and filtered. INSCoV-501B (8.64 mg, 16.26 μmol, 2.59% yield, 94.862% purity) was obtained as an orange solid, which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0657] LCMS: retention time: 0.581 min, (M+H)=504.0, HPLC: retention time: 1.332 min, SFC: retention time: 1.718 min, 2.024 min, 1 H NMR (400MHz, DMSO-d6): δ = 8.98 (s, 1H), 8.51 (s, 2H), 8.46 (s, 1H), 8.43 (d, J = 7.8Hz, 1H), 7.72 (s, 1H), 7.66-7.64 (m, 2H), 7.42-7.41 (m ,1H),6.04(s,1H),4.10-3.96(m,3H),3.28-3.19(m,2H),3.13-3.04(m,1H),3.03-2.94(m,1H),2.12-1.90(m,3H),1.86-1.73(m,1H).

[0658] Example 21. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyrimidin-5-yl)-2-(1-methyl Synthesis of (phenylsulfonylethyl)amino)ethyl)acetamide (INSCoV-501C(2))

[0659] INSCoV-501C (2) was synthesized according to the general procedure for the INSCoV series and purified by the following purification method A: the residue was dissolved in MeOH (2 mL) and purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 32%-52%, 6.5 min) and concentrated to remove MeCN. The liquid was lyophilized to give the product. INSCoV-501C (2) (28.59 mg, 47.35 μmol, 5.12% yield, 91.759% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0660] LCMS: retention time: 0.831 min, (M+H)=554.2, HPLC: retention time: 1.982 min, SFC: retention time: 1.802 min, 2.474 min, 1 H NMR (400MHz, DMSO-d6): δ=9.32(d,J=9.2Hz,1H),8.94(s,1H),8.47-8.43(m,1H),8.34(s,2H),7.80(d,J=8.2Hz,2H),7.75-7.70( m,1H),7.60(d,J=6.8Hz,2H),7.49-7.26(m,3H),6.21(s,1H),5.25-5.19(m,1H),3.94(s,2H),2.42(s,3H),1.35(d,J=7.0Hz,3H).

[0661] Example 22. Synthesis of INSCoV-501G

[0662] Plan 19

[0663]

[0664] Step 1: To a solution of compound 1 (3 g, 18.17 mmol, 1 eq) and compound 2 (4.26 g, 21.80 mmol, 1.2 eq) in MeOH (70 mL) was added K2CO3 (5.02 g, 36.33 mmol, 2 eq). At 70 ° C, under N2, the reaction mixture was stirred for 1 h. TLC (PE: EA = 4: 1) showed that compound 1 (Rf = 0.8) was completely consumed and two new spots (Rf = 0.4, Rf = 0.0) were formed. The reaction mixture was concentrated in vacuo. The residue was diluted with NaHCO3 solution (20 mL) and extracted with EA (30 mL × 2). The combined organic phase was washed with water (30 mL), dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 80g The mixture was purified by silica flash column, 0-50% ethyl acetate / petroleum ether gradient eluent at 100 mL / min and concentrated in vacuo to obtain compound 3 (3.5 g, 16.93 mmol, 93.22% yield, 98.79% purity) as a yellow solid, which was confirmed by LCMS and HNMR.

[0665] LCMS: retention time: 0.930 min, (M+H)=205.2. 1H NMR (400MHz, DMSO-d6) δ = 8.58 (d, J = 1.2Hz, 1H), 8.11 (d, J = 8.6Hz, 1H), 7.94 (d, J = 1.2Hz, 1H), 7.88 (s, 1H), 7.79 (d, J = 8.8Hz, 1H), 2.58 (s, 3H).

[0666] Step 2: Pd / C (1.5 g, 7.35 mmol, 10% purity, 1 eq) was added to a solution of compound 3 (1.5 g, 7.35 mmol, 1 eq) in MeOH (15 mL). The reaction mixture was stirred for 12 h at 25 ° C under H2 (15 PSI). LCMS showed that compound 3 was completely consumed, and a peak with the desired mass was detected (Rt = 0.827 min). The reaction mixture was filtered through a celite pad and washed with MeOH (20 mL × 2). The filtrate was concentrated in vacuo. The reaction was used for the next step without purification. Compound 4 (1 g, crude product) was obtained as an off-white solid.

[0667] Step 3: INSCoV-501G was obtained according to the general procedure for the INSCoV series and purified by the following purification B: MTBE (20 mL) was added to the reaction mixture and cooled to 0 ° C for 1 h. The mixture was filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-501G (192.57 mg, 409.06 μmol, 29.48% yield, 99.403% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0668] LCMS: Retention time: 0.933 min, (M+H) = 468.3. HPLC: Retention time: 2.126 min. SFC: Retention time: 1.002 min, 1.136 min. 1 H NMR (400MHz, DMSO-d6) δ = 8.93 (s, 1H), 8.48 (s, 2H), 8.45 (s, 1H), 7.99 (d, J = 8 .4Hz,1H),7.74-7.67(m,1H),7.67-7.61(m,1H),7.42(d,J=1.6Hz,1H),6.04( s,1H),4.02-3.86(m,2H),3.63-3.51(m,1H),1.89(s,3H),1.83-1.74(m,1H) ,1.72-1.65(m,1H),1.60-1.48(m,3H),1.33-1.12(m,4H),0.99-0.87(m,1H).

[0669] Example 23. Synthesis of INSCoV-501H

[0670] Plan 20

[0671]

[0672] Step 1: At 0°C, to a solution of compound 1 (40 g, 199.72 mmol, 1 eq) and DIPEA (77.44 g, 599.17 mmol, 104.36 mL, 3 eq) in DCM (600 mL) was added compound 2 (33.69 g, 239.67 mmol, 1.2 eq). The mixture was stirred at 20°C for 16 h. TLC (PE:EA=1:1) showed that compound 1 (Rf=0.05) was consumed, and a new spot (Rf=0.5) was observed. The mixture was poured into saturated NaHCO (400 mL) and then extracted with DCM (200 mL×2). The combined organic layers were washed with water (200 mL), brine (100 mL), dried over NaSO, filtered, and concentrated in vacuo to yield a residue. The residue was triturated with MTBE (400 mL). The filter cake was washed with MTBE (100 mL x 2) and then dried in vacuo to obtain compound 3 (56 g, 183.97 mmol, 92.11% yield) as a white solid, which was identified by HNMR.

[0673] 1 H NMR (400MHz, chloroform-d): δ = 4.59-4.37 (m, 1H), 3.76 (br d, J = 12.2Hz, 2H), 3.57 (br s, 1H), 2.92 (br t,J=11.1Hz,2H),2.26(tt,J=4.8,8.0Hz,1H),2.10-1.96(m,2H),1.56-1.38(m,11H),1.21-1.11(m,2H),1.04-0.93(m,2H).

[0674] Step 2: At 0 ° C, TFA (15 mL) was added to a solution of compound 3 (10 g, 32.85 mmol, 1 eq) in DCM (100 mL). The reaction was stirred for 5 h at 25 ° C. TLC (PE: EA = 3: 1, I2) indicated that compound 3 (Rf = 0.3) was completely consumed and three new spots (Rf = 0.05, Rf = 0.6, Rf = 0.8) were formed. At 25 ° C, the reaction mixture was added to 100 mL of H2O and extracted with DCM (100 mL × 2). The pH of the aqueous phase was adjusted to 9 by saturated NaHCO3, and it was extracted with DCM (100 mL × 6). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to produce a crude product. The crude product was used in the next step without further purification. Compound 4 (1 g, 4.90 mmol, 14.90% yield) was obtained as a white solid, which was verified by HNMR.

[0675] 1 H NMR (400 MHz, chloroform-d): δ = 3.69 (td, J = 3.2, 12.4 Hz, 2H), 2.90-2.70 (m, 3H), 2.19 (tt, J = 4.8, 8.0 Hz, 1H), 1.55-1.30 (m, 6H), 1.12-1.03 (m, 2H), 0.94-0.84 (m, 2H).

[0676] Step 3: At 25 ° C, a mixture of HCOOH (261.02 mg, 5.43 mmol, 3.0 eq) and Ac2O (221.88 mg, 2.17 mmol, 203.56 μ L, 1.2 eq) was stirred for 10 min. A solution of compound 4 (370 mg, 1.81 mmol, 1 eq) in DCM (5 mL) was slowly added to the above mixture. The mixture was stirred for 15 h at 25 ° C. TLC (MeOH: DCM = 1: 10, I2) showed that compound 4 (Rf = 0) was consumed, and a new spot (Rf = 0.3) was observed. At 10 ° C, the mixture was slowly poured into saturated NaHCO3 (100 mL), then extracted with DCM (80 mL × 2). The combined organic layer was washed with brine (40 mL), dried over Na2SO4, filtered and concentrated in vacuo to produce a crude product. The crude product was used in the next step without further purification. Compound 5 (370 mg, 1.59 mmol, 87.94% yield) was obtained as a light yellow solid, which was verified by HNMR.

[0677] 1H NMR (400 MHz, chloroform-d): δ = 8.23-8.05 (m, 1H), 5.73 (br s, 1H), 4.14-3.97 (m, 1H), 3.89-3.74 (m, 2H), 3.09-2.84 (m, 2H), 2.37-2.20 (m, 1H), 2.12-2.00 (m, 2H),

[0678] Step 4: At 0 ° C, to a solution of compound 5 (350 mg, 1.51 mmol, 1 eq) and Et3N (457.38 mg, 4.52 mmol, 629.14 μ L, 3.0 eq) in DCM (30 mL) was slowly added dropwise POCl3 (693.06 mg, 4.52 mmol, 420.03 μ L, 3.0 eq). The mixture was stirred at 0 ° C for 1 h. TLC (DCM: MeOH = 10: 1) showed that compound 5 (Rf = 0) remained, and a new spot (Rf = 0.3) was observed. At 10 ° C, the mixture was slowly poured into saturated NaHCO3 (60 mL) and then extracted with DCM (45 mL × 2). The combined organic layers were washed with brine (20 mL × 2), dried over Na2SO4, filtered and concentrated in vacuo to produce a crude product. The crude product was purified by silica gel chromatography, eluting with petroleum ether / ethyl acetate = 1: 1. Compound 6 (220 mg, 1.03 mmol, 68.14% yield) was obtained as a yellow solid, which was verified by HNMR.

[0679] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 3.93 (br s, 1H), 3.56-3.45 (m, 2H), 3.44-3.29 (m, 2H), 2.28 (tt, J = 4.9, 8.0 Hz, 1H), 2.03-1.91 (m, 4H), 1.23-1.14 (m, 2H), 1.07-0.97 (m, 2H).

[0680] Step 5: INSCoV-501H was obtained according to the general procedure for the INSCoV series and purified by the following purification B: the reaction was concentrated in vacuo. The crude product was dissolved in ethyl acetate (6 mL), stirred for a moment and filtered, and the filter cake was concentrated in vacuo. INSCoV-501H (102.99 mg, 174.30 μmol, 37.35% yield, 94.609% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS and HPLC.

[0681] LCMS: Retention time: 0.793 min, (M+H) = 559.1 / 561.1; Retention time: 0.787 min, (M+H) = 559.1 / 561.1. HPLC: Retention time: 1.784 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.98 (s, 1H), 8.51 (s, 2H), 8.46 (s, 1H), 8.38 (d, J = 7.5Hz, 1H), 7.72 (s, 1H), 7.66 (br d,J=8.4Hz,2H),7.53-7.26(m,2H),6.10(s,1H),4.10-4.00(m,2H),3.86-3.75(m,1H),3.60-3.46(m,2H),3.01-2.8 7(m,2H),2.62-2.52(m,2H),1.93-1.73(m,2H),1.57-1.40(m,1H),1.32(dt,J=8.3,11.2Hz,1H),1.00-0.89(m,4H).

[0682] Example 24. Synthesis of INSCoV-501H (1)

[0683] Plan 21

[0684]

[0685] Step 1: At 0 ° C, to a solution of compound 1 (10 g, 49.93 mmol, 1 eq) and Et3N (7.58 g, 74.90 mmol, 10.42 mL, 1.5 eq) in DCM (60 mL) was added ethanesulfonyl chloride (7.06 g, 54.92 mmol, 5.19 mL, 1.1 eq). The mixture was stirred at 25 ° C for 1 h. TLC (PE:EA=1:1, ninhydrin) indicated complete consumption of compound 1 (Rf=0.2) and the formation of two new spots (Rf=0.7, Rf=0.4). At 0 ° C, the reaction mixture was quenched by adding 100 mL of H2O and extracted with DCM (100 mL×3). The combined organic layers were washed with brine (40 mL×3), dried over Na2SO4, filtered and concentrated under reduced pressure to produce a residue. The crude product was triturated with EA (100 mL) at 25° C. for 20 min, then filtered to give a white solid. Compound 2 (12 g, 41.04 mmol, 82.20% yield) was obtained as a white solid, which was verified by HNMR.

[0686] 1H NMR (400MHz, chloroform-d): δ = 4.68-4.29 (m, 1H), 3.78 (br d, J = 12.3Hz, 2H), 3.68-3.50 (m, 1H), 3.01-2.87 (m, 4H), 2.03 (br dd,J=2.7,12.8Hz,2H),1.55-1.49(m,1H),1.55-1.48(m,1H),1.46(s,9H),1.37(t,J=7.5Hz,3H).

[0687] Step 2: at 0 ° C, TFA (7.5 mL) was added to a solution of compound 2 (8 g, 27.36 mmol, 1 eq) in DCM (50 mL). The reaction was stirred for 5 h at 25 ° C. TLC (PE: EA = 1: 1, ninhydrin) indicated that compound 2 was completely consumed and a new spot was formed. At 0 ° C, the reaction mixture was quenched by adding 80 mL of H2O. The reaction mixture was extracted with DCM (100 mL × 10). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to produce a residue. Compound 3 (3.5 g, 18.20 mmol, 66.53% yield) was obtained as a light yellow oil, which was verified by HNMR.

[0688] 1 H NMR (400 MHz, chloroform-d): δ = 3.65 (br dd, J = 2.8, 12.2 Hz, 2H), 2.96-2.69 (m, 5H), 1.88-1.74 (m, 2H), 1.57-1.42 (m, 2H), 1.39-1.21 (m, 4H), 1.22-1.12 (m, 1H).

[0689] Step 3: At 25 ° C, a mixture of HCOOH (1.50 g, 31.20 mmol, 3.0 eq) and Ac2O (1.27 g, 12.48 mmol, 1.17 mL, 1.2 eq) was stirred for 10 min. A solution of compound 3 (2.0 g, 10.40 mmol, 1 eq) in DCM (20 mL) was slowly added to the above mixture. The mixture was stirred for 3 h at 25 ° C. TLC (MeOH: DCM = 1: 10, I2) showed that compound 3 (Rf = 0) was consumed, and a new spot (Rf = 0.3) was observed. At 10 ° C, the mixture was slowly poured into saturated NaHCO3 (100 mL), then extracted with DCM (80 mL × 2). The combined organic layer was washed with brine (40 mL), dried over Na2SO4, filtered and concentrated in vacuo to produce a crude product. The crude product was used in the next step without further purification. Compound 4 (2.25 g, 10.21 mmol, 98.20% yield) was obtained as a light yellow solid, which was verified by HNMR.

[0690] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 8.15 (s, 1H), 5.82-5.56 (m, 1H), 4.17-3.94 (m, 1H), 3.83 (br d, J = 12.8 Hz, 2H), 3.05-2.82 (m, 4H), 2.12-1.99 (m, 2H), 1.62-1.49 (m, 2H), 1.37 (t, J = 7.4 Hz, 3H).

[0691] Step 4: At 0 ° C, to a solution of compound 4 (0.3 g, 1.36 mmol, 1 eq) and Et3N (413.41 mg, 4.09 mmol, 568.66 μ L, 3.0 eq) in DCM (30 mL) was slowly added dropwise POCl3 (626.45 mg, 4.09 mmol, 379.66 μ L, 3.0 eq). The mixture was stirred for 1 h at 0 ° C. TLC (PE: EA = 1: 1, I2) showed that compound 4 (Rf = 0) was consumed, and a new spot (Rf = 0.45) was observed. At 10 ° C, the mixture was slowly poured into saturated NaHCO3 (100 mL) and then extracted with DCM (60 mL × 2). The combined organic layers were washed with brine (30 mL × 2), dried over Na2SO4, filtered and concentrated in vacuo to produce a crude product. The crude product was purified by silica gel chromatography eluting with petroleum ether / ethyl acetate=1:1 to give the desired product. Compound 5 (165 mg, 815.73 μmol, 59.90% yield) was obtained as a colorless oil, which was verified by HNMR.

[0692] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 3.94 (br s, 1H), 3.63-3.46 (m, 2H), 3.46-3.30 (m, 2H), 2.98 (q, J = 7.4 Hz, 2H), 2.04-1.89 (m, 4H), 1.38 (t, J = 7.4 Hz, 3H).

[0693] Step 5: INSCoV-501H (1) was obtained according to the general procedure for the INSCoV series and subjected to the following purification B: The reaction was concentrated in vacuo. The crude product was dissolved in ethyl acetate (2 mL), stirred for a while and filtered, and the filter cake was concentrated in vacuo. INSCoV-501H (1) (83.93 mg, 144.18 μmol, 46.19% yield, 93.968% purity) was obtained as an orange solid, which was confirmed by LCMS, HPLC and HNMR.

[0694] LCMS: retention time: 0.652 min, (M+H) = 547.1; retention time: 0.790 min, (M+H) = 547.1 HPLC: retention time: 1.752 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.97 (s, 1H), 8.49 (s, 2H), 8.46 (s, 1H), 8.36 (d, J = 7.5Hz, 1H), 7.73-7.70 (m, 1H), 7.65 (br d,J=8.6Hz,2H),7.49-7.33(m,2H),6.08(s,1H),4.05(br d,J=6.2Hz,2H),3.84-3.74(m,1H),3.60-3.39(m,3H),3.07-3.01(m,2H),2.95-2.86 (m,2H),1.88-1.74(m,2H),1.50-1.40(m,1H),1.33-1.25(m,1H),1.21-1.17(m,3H).

[0695] Example 25. 2-Chloro-N-(2-((1,1-dioxotetrahydro-2H-thiopyran-4-yl)amino)-2-oxo-1-(pyrrolidone) Synthesis of N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501M)

[0696] INSCoV-501M was synthesized according to the general procedure for the INSCoV series and the following purification was performed A: The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150×25 mm×10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 18%-48%, 11 min) and dried by lyophilization. INSCoV-501M (9.55 mg, 18.40 μmol, 6.63% yield, 97.122% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0697] LCMS: Retention time: 0.807 min, (M+H) = 504.3; Retention time: 0.808 min, (M+H) = 504.2. HPLC: Retention time: 1.351 min. SFC: Retention time: 1.815 min, 2.285 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.74-8.33 (m, 5H), 8.13 (s, 1H), 7.69 (s, 1H), 7.65-7. 36(m,4H),6.23(s,1H),4.28-3.62(m,3H),3.12-2.92(m,2H),2.10-1.81(m,4H).

[0698] Example 26. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyrazin-2-yl)-2-(toluene Synthesis of (Sulphonylmethyl)amino)ethyl)acetamide (INSCoV-501O)

[0699] INSCoV-501O was obtained according to the general procedure for the INSCoV series and subjected to the following purification A: The residue was dissolved in MeOH (2 mL) and purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 30%-50%, 6.5 min) and concentrated to remove MeCN. The liquid was lyophilized to produce the product. INSCoV-501O (274.41 mg, 486.27 μmol, 52.56% yield, 95.689% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0700] LCMS: retention time: 0.873 min, (M+H) = 540.1; retention time: 0.864 min, (M+H) = 540.1, HPLC: retention time: 1.830 min. SFC: retention time: 0.846 min, 1.337 min. 1H NMR (400MHz, DMSO-d6): δ=9.50-9.32(m,1H),8.51-8.45(m,1H),8.46-8.40(m,2H),8.25(s,1H),7.73-7.63( m,3H),7.60-7.54(m,2H),7.40-7.26(m,3H),6.26(s,1H),4.93-4.63(m,2H),4.14-3.93(m,2H),2.39(s,3H).

[0701] Example 27. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-2-(phenethylamino)-1-(pyrrolidone)- Synthesis of (2-oxazine-2-yl)ethyl)acetamide (INSCoV-501P)

[0702] INSCoV-501P was obtained according to the general procedure of the INSCoV series. The residue was diluted with MeOH (4 mL), purified by preparative HPLC (column: 3-Phenomenex Luna C18 75 × 30 mm × 3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 34%-54%, 6.5 min) and concentrated to remove MeCN, and the liquid was lyophilized to produce the product. INSCoV-501P (89.34 mg, 174.23 μmol, 18.83% yield, 92.815% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0703] LCMS: Retention time: 0.879 min, (M+H) = 476.2; Retention time: 0.883 min, (M+H) = 476.1. HPLC: Retention time: 1.913 min. SFC: Retention time: 0.786 min, 1.306 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.50-8.37 (m, 4H), 7.70 (s, 1H), 7.60 (d, J = 8.4Hz, 2H), 7.49-7.35 (m ,1H),7.31-7.10(m,6H),6.19(s,1H),4.18-3.97(m,2H),3.45-3.26(m,2H),2.77-2.63(m,2H).

[0704] Example 28. 2-Chloro-N-(2-((1-(cyclopropylsulfonyl)piperidin-4-yl)amino)-2-oxo-1-(pyrazine- Synthesis of N-(2-(4-(oxazol-5-yl)ethyl)-N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501R)

[0705] INSCoV-501R was obtained according to the general procedure of the INSCoV series. Purification B: The reaction was concentrated in vacuo. The crude product was dissolved in ethyl acetate (4 mL), stirred for a moment and filtered, and the filter cake was concentrated in vacuo. INSCoV-501R (139.16 mg, 239.21 μmol, 51.26% yield, 96.095% purity) was obtained as an off-white solid, which was determined by LCMS, HPLC and HNMR.

[0706] LCMS: retention time: 0.793 min, (M+H) = 547.1; retention time: 0.780 min, (M+H) = 547.1 HPLC: retention time: 1.805 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.56-8.48 (m, 2H), 8.46-8.41 (m, 2H), 8.35 (d, J = 7.4Hz, 1H), 7.70 (s, 1H), 7.61 (br d,J=8.5Hz,2H),7.57-7.28(m,2H),6.24(s,1H),6.30-6.19(m,1H),4.18-4.09(m,1H),4.09-4. 00(m,1H),3.81-3.69(m,1H),3.53-3.43(m,2H),3.00-2.88(m,2H),2.58-2.51(m,2H),1.79(br d,J=11.7Hz,2H),1.51-1.30(m,2H),0.97-0.84(m,4H).

[0707] Example 29. 2-Chloro-N-(2-((1-(ethylsulfonyl)piperidin-4-yl)amino)-2-oxo-1-(pyrazine-2-yl)amino)-2-nitro-1-nitro ...2-nitro-1-nitro-2-nitro Synthesis of N-(4-(oxazol-5-yl)ethyl)-N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501R(1))

[0708] INSCoV-501R (1) was obtained according to the general procedure for the INSCoV series. Purification B: The reaction was concentrated in vacuo. The crude product was dissolved in ethyl acetate (2 mL), stirred for a while and filtered, and the filter cake was concentrated in vacuo. INSCoV-501R (1) (73.31 mg, 131.92 μmol, 42.26% yield, 98.439% purity) was obtained as an orange solid, which was confirmed by LCMS, HPLC and HNMR.

[0709] LCMS: Retention time: 0.642 min, (M+H) = 547.1; Retention time: 0.780 min, (M+H) = 547.1. HPLC: Retention time: 1.735 min. 1H NMR (400MHz, DMSO-d6): δ=8.53-8.47(m,2H),8.45-8.42(m,2H),8.33(d,J=7.6Hz,1H),7.69(s,1H),7.60(br d,J=8.4Hz,2H),7.46(br d,J=2.4Hz,2H),6.23(s,1H),4.17-4.10(m,1H),4.07-3.99(m,1H),3.81-3.71(m,1H),3.48(br t,J=13.3Hz,2H),3.08-2.99(m,2H),2.95-2.88(m,2H),1.82-1.73(m,2H),1.40-1.26(m,2H),1.21-1.14(m,4H).

[0710] Example 30. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrazin-2-yl)ethyl)- Synthesis of N-(4-(oxazol-5-yl)phenyl)acetamide (INSCoV-501S)

[0711] INSCoV-501S was obtained according to the general procedure for the INSCoV series. Purification A: The reaction mixture was concentrated in vacuo to give a residue. The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150×25 mm×10 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 35%-65%, 10 min) to give INSCoV-501S (100 mg, 204 μmol, 29% yield, 99.9% purity) as a yellow solid, which was confirmed by LCMS, HPLC, and HNMR.

[0712] LCMS: retention time: 0.842 min, (M+H) = 490.4. HPLC: retention time: 1.796 min. 1 H NMR (400MHz, CDCl3): δ=8.75(s,1H),8.54(m,1H),8.51(m,1H),7.95(s,1H),7.67(s,1H),7.65(s,1H),7.45-7.44(m,2H),7 .41(s,1H),7.33(d,J=7.6Hz,1H),5.89(s,1H),3.98-3.87(m,3H),2.10-1.97(m,3H),1.94-1.79(m,3H),1.63-1.47(m,2H).

[0713] Example 31. 2-Chloro-N-(3-chloro-4-methoxyphenyl)-N-(2-(cyclohexylamino)-2-oxo-1-(pyrrolidone)-1-yl)-2-nitro-1-nitro-2 ... Synthesis of (3-pyridin-3-yl)ethyl)acetamide (INSCoV-509)

[0714] INSCoV-509 was obtained according to the general procedure for the INSCoV series. Purification A: The residue was purified by preparative HPLC (column: Phenomenex Luna C18 150×25mm×10um; mobile phase: [water (0.225% FA)-ACN]; B%: 36%-66%, 10min) to obtain a solution. The crude product was purified by preparative HPLC (column: Phenomenex lunaC18 150×25mm×10um; mobile phase: [water (0.225% FA)-ACN]; B%: 36%-66%, 10min) to obtain solution 2. Solution 1 and solution 2 were combined and dried by lyophilization. INSCoV-509 (162.68 mg, 353.59 μmol, 27.86% yield, 97.886% purity) was obtained as a white solid, which was confirmed by LCMS, HPLC and HNMR.

[0715] LCMS: Retention time: 0.696 min, [M+H] = 450.0; Retention time: 0.858 min, [M+H] = 450.2. HPLC: Retention time: 1.882 min. 1 H NMR (400MHz, DMSO-d6): δ=8.40-8.24(m,2H),8.14(d,J=7.2Hz,1H),8.00-7.54(m,1H),7.41-7.30(m,1H),7.23-7.13(m,1H),7.1 1-6.68(m,2H),6.10-5.97(m,1H),4.09-3.90(m,2H),3.86-3.71(m,3H),3.66-3.52(m,1H),1.79-1.49(m,5H),1.31-0.94(m,5H).

[0716] Example 32. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(2,4-dichloro- Synthesis of (Methoxyphenyl)acetamide (INSCoV-512)

[0717] INSCoV-512 was obtained according to the general procedure for the INSCoV series. Purification A: The crude product was purified by reverse phase HPLC (0.1% FA conditions) and concentrated in vacuo to remove MeCN and dried by lyophilization. INSCoV-512 (255.93 mg, 553.41 μmol, 59.28% yield, 96.427% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0718] LCMS: Retention time: 0.737 min, [M+H] = 446.0; Retention time: 0.831 min, [M+H] = 446.1. HPLC: Retention time: 1.682 min. SFC: Retention times: 1.143 min and 1.441 min. 1 H NMR (400MHz, DMSO-d6): δ=8.28-8.27(m,1H),8.20(d,J=1.8Hz,1H),7.99(d,J=7.8H z,1H),7.61(d,J=8.8Hz,1H),7.30-7.28(m,1H),7.10-7.08(m,1H),6.48-6.45(m,1H ),6.23(d,J=2.8Hz,1H),5.88(s,1H),3.96(d,J=14.2Hz,1H),3.79(d,J=14.2Hz,1H) ,3.67(s,3H),3.60-3.52(m,1H),3.45(s,3H),1.79-1.52(m,5H),1.25-0.93(m,5H).

[0719] Example 33. Synthesis of INSCoV-517B

[0720] Plan 22

[0721]

[0722] Step 1: at 0 ° C, under N2, to a solution of compound 1 (500 mg, 2.47 mmol, 1 eq) and compound 2 (267.39 mg, 2.47 mmol, 1 eq) in DCM (0.5 mL) was added TiCl4 (1 M, 1.24 mL, 0.5 eq), TEA (750.91 mg, 7.42 mmol, 1.03 mL, 3 eq). The mixture was stirred for 1 h at 0 ° C. The mixture was then warmed to 30 ° C and stirred for 11 h. LCMS showed that compound 1 was consumed and the required mass was detected. The mixture was dissolved in DCM (50 mL) and quenched with saturated NH4Cl (50 mL), separated, and the aqueous phase was extracted with DCM (3 × 20 mL). The organic layer was washed with brine, dried over anhydrous Na2SO4, filtered and concentrated in vacuo. Compound 3 (210 mg, crude product) was obtained as a yellow oil. It was used directly in the next step.

[0723] LCMS: retention time: 0.961 / 0.989 min, (M+H)=293.1.

[0724] Step 2: prepare INSCoV-517B by the general method for INSCoV series. Purification B: the mixture is concentrated in vacuo. The residue is dissolved in METB (8 mL), stirred for 15 min, filtered, and the filter cake is dissolved in ethyl acetate (8 mL), stirred for 15 min, filtered, and the filter cake is concentrated in vacuo. INSCoV-517B (65.13 mg, 115.00 μmol, 16.69% yield, 93.909% purity) as an off-white solid is obtained. It is confirmed by HNMR, FNMR, LCMS and HPLC.

[0725] LCMS: Retention time: 1.000 min, (M+H) = 532.3; Retention time: 0.903 min, (M+H) = 532.3. HPLC: Retention time: 2.219 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.75-8.60 (m, 2H), 8.48 (d, J = 2.4Hz, 1H), 8.39-8.33 (m, 1H), 8.23 ​​(br d, J = 3.5Hz, 1H), 8.13 (br s,1H),7.88-7.72(m,2H),6.32(s,1H),4.14(br d,J=2.1Hz,2H),3.87-3.57(m,1H),2.04-1.62(m,8H),1.53-1.27(m,2H). 19 F NMR (377MHz, DMSO-d6): δ=-93.00--94.10(m,1F),-96.71--98.22(m,1F). 1 HNMR (400MHz, DMSO-d6, T=50): δ=8.60-8.52(m,1H),8.45-8.39(m,1H),8.31(br s,1H),8.22-8.14(m,1H),8.13-8.08(m,1H),8.06-7.95(m,1H),7.79-7.66(m,2H),6. 29(s,1H),4.20-3.98(m,2H),3.82-3.70(m,1H),1.99-1.65(m,8H),1.50-1.30(m,2H). 19 F NMR (376MHz, DMSO-d6, T=50): δ=-93.35--94.20(m,1F),-96.66--97.90(m,1F).

[0726] Example 34. N-(tert-Butyl)-2-(N-(4-(tert-Butyl)phenyl)-2-chloroacetamido)-2-(pyridin-3-yl) Synthesis of acetamide (INSCoV-534)

[0727] INSCoV-534 was obtained according to the general procedure for the INSCoV series. Purification A: The residue was purified by preparative HPLC (column: Phenomenex luna C18 150×40 mm×15 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 37%-67%, 10 min). N-tert-butyl-2-(4-tert-butyl-N-(2-chloroacetyl)anilino)-2-(3-pyridyl)acetamide (50.79 mg, 121.23 μmol, 11.46% yield, 99.286% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0728] HPLC: Retention time: 1.912 min. LCMS: Retention time: 0.886 min, (M+H) = 416.4; Retention time: 0.870 min, (M+H) = 416.3. 1 H NMR (400MHz, DMSO-d6): δ=8.33-8.24(m,2H),7.84(s,1H),7.38-7.01(m,5H),6.00(s,1H),4.00-3.89(m,2H),1.21(s,9H),1.17(s,9H).

[0729] Example 35. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-1-(5-hydroxypyridine-3-yl)- Synthesis of 2-(2-(2-methyl)-2-oxoethyl)acetamide (INSCoV-535)

[0730] INSCoV-535 was obtained according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: Phenomenex luna C18 150×40 mm×15 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 30%-60%, 10 min) and preparative HPLC (column: Welch MLtimate XB-SiOH 250×50×10 μm; mobile phase: [hexane-EtOH (0.1% NH3·H2)]; B%: 1%-30%, 15 min). 2-(4-tert-Butyl-N-(2-chloroacetyl)anilino)-N-cyclohexyl-2-(5-hydroxy-3-pyridinyl)acetamide (16.39 mg, 33.43 μmol, 3.16% yield, 93.404% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0731] HPLC: Retention time: 2.238 min. LCMS: Retention time: 0.799 min, (M+H) = 458.3; Retention time: 0.883 min, (M+H) = 458.1. 1H NMR (400MHz, DMSO-d6): δ = 9.71 (s, 1H), 8.04 (br d, J = 7.2Hz, 1H), 7.85 (d, J = 2.4Hz, 1H), 7.77 (d, J = 1.2Hz, 1H), 7.24 (br d,J=6.0Hz,4H),6.71(s,1H),5.95(s,1H),4.01-3.85(m,2H),3.63-3.50(m,1H),1.79-1.48(m,5H),1.28-1.05(m,14H).

[0732] Example 36. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyrimidine-5- Synthesis of 2-(4-amino)ethyl)acetamide (INSCoV-536)

[0733] INSCoV-536 was obtained according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 57%-87%, 7 min). INSCoV-536 (45.49 mg, 97.79 μmol, 9.24% yield, 95.224% purity) was obtained as a yellow solid, which was confirmed by LCMS, HPLC and HNMR.

[0734] HPLC: retention time: 2.451 min. LCMS: retention time: 0.998 min, (M+H)=443. 1 H NMR (400MHz, DMSO-d6): δ = 8.95 (s, 1H), 8.43 (s, 2H), 8.13 (br d,J=7.2Hz,1H),7.39-7.11(m,4H),6.03(s,1H),4.06-3.91(m,2H),3.65-3.49(m,1H),1.78-1.54(m,5H),1.35-1.13(m,14H).

[0735] Example 37. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridazine-4- Synthesis of 2-(4-amino)ethyl)acetamide (INSCoV-537)

[0736] INSCoV-537 was obtained according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: Phenomenex luna C18 150×40 mm×15 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 45%-75%, 10 min) and preparative HPLC (column: Welch MLtimate XB-CN 250×70×10 μm; mobile phase: [hexane-EtOH (0.1% NH 3 · H 2 O)]; B%: 10%-50%, 15 min) to obtain INSCoV-537 (15.87 mg, 34.62 μmol, 3.27% yield, 96.637% purity) as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0737] HPLC: retention time: 2.595 min. LCMS: retention time: 0.973 min, (M+H)=443.2 。 1 H NMR (400MHz, DMSO-d6): δ = 9.07-9.02 (m, 1H), 9.00 (s, 1H), 8.18 (br d,J=7.2Hz,1H),7.40-7.20(m,5H),5.99(s,1H),4.09-3.94(m,2H),3.59-3.48(m,1H),1.64(br d,J=4.8Hz,4H),1.53(br d,J=12.0Hz,1H),1.21(s,10H),1.16-1.00(m,3H).

[0738] Example 38. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyridazin-4-yl)

[0739] Synthesis of (4-(oxazol-5-yl)phenyl)acetamide (INSCoV-537I)

[0740] INSCoV-537I was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was ground with EtOH (2 ml), filtered, and the filter cake was washed with PE (5 ml) and dried in vacuo. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyridazin-4-yl)ethyl)-N-(4-(oxazol-5-yl)phenyl)acetamide (194.18 mg, 364.45 μmol, 43.05% yield, 91.948% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, and HPLC.

[0741] HPLC: Retention time: 2.064 min. LCMS: Retention time: 0.771 min, (M+H) = 490.2; Retention time: 0.842 min, (M+H) = 490.0. 1 H NMR (400MHz, DMSO-d6): δ = 9.15-8.98 (m, 2H), 8.46 (s, 1H), 8.38 (br d,J=6.8Hz,1H),7.79-7.60(m,1H),7.81-7.57(m,2H),7.55-7.32(m,1H),7.57-7.31(m,1H ),6.06(s,1H),4.20-3.99(m,2H),3.94-3.75(m,1H),2.13-1.68(m,7H),1.60-1.30(m,2H).

[0742] Example 39. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyridazin-4-yl)ethyl)- Synthesis of N-(4-(Isoxazol-5-yl)phenyl)acetamide (INSCoV-537K)

[0743] INSCoV-537K was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was triturated with EtOH (2 ml), filtered, and the filter cake was washed with PE (5 ml) and dried under vacuum. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyridazin-4-yl)ethyl)-N-(4-(oxazol-5-yl)phenyl)acetamide (194.18 mg, 364.45 μmol, 43.05% yield, 91.948% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, and HPLC.

[0744] HPLC: Retention time: 2.064 min. LCMS: Retention time: 0.771 min, (M+H) = 490.2; Retention time: 0.842 min, (M+H) = 490.0. 1 H NMR (400MHz, DMSO-d6): δ = 9.15-8.98 (m, 2H), 8.46 (s, 1H), 8.38 (br d,J=6.8Hz,1H),7.79-7.60(m,1H),7.81-7.57(m,2H),7.55-7.32(m,1H),7.57-7.31(m,1H ),6.06(s,1H),4.20-3.99(m,2H),3.94-3.75(m,1H),2.13-1.68(m,7H),1.60-1.30(m,2H).

[0745] Example 40. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyrazine-2-yl)- Synthesis of (4-amino)ethyl)acetamide (INSCoV-538)

[0746] INSCoV-538 was synthesized according to the general procedure for the INSCoV series. Purification A: The crude product was triturated with MTBE (5 ml), filtered, the filter cake was triturated with EtOH (2 ml), filtered, and the filter cake was dried under vacuum. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyrazin-2-yl)ethyl)acetamide (272.7 mg, 599.17 μmol, 56.62% yield, 97.330% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS, and HPLC.

[0747] HPLC: retention time: 2.804 min. LCMS: retention time: 1.029 min, (M+H)=443.2, retention time: 1.014 min, (M+H)=443.2. 1 H NMR (400MHz, DMSO-d6): δ = 8.58-8.36 (m, 3H), 8.14 (br d, J = 7.2Hz, 1H), 7.28 (br s,4H),6.13(s,1H),4.13-3.92(m,2H),3.58-3.42(m,1H),1.73-1.44(m,5H),1.21(s,11H),1.13-0.97(m,3H).

[0748] Example 41. N-(4-(tert-butyl)phenyl)-2-chloro-N-(1-(5-cyanopyridin-3-yl)-2-(cyclohexylamino)- Synthesis of 2-(2-(2-methyl)-2-oxoethyl)acetamide (INSCoV-539)

[0749] INSCoV-539 was synthesized according to the general procedure for the INSCoV series. Purification B: The mixture was poured into EtOH (5 ml). The mixture was filtered. The filter cake was washed with EtOH (1 ml) and MTBE (1 ml) and dried under vacuum. INSCoV-539 (309.39 mg, 648.42 μmol, 61.27% yield, 97.874% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0750] HPLC: retention time: 2.895 min. LCMS: retention time: 1.029 min, (M+H)=467.1. 1H NMR (400MHz, DMSO-d6): δ=8.84(d,J=1.8Hz,1H),8.57(d,J=2.0Hz,1H),8.13(d,J=8.0Hz,1H),7.75(t,J=2.0H z,1H),7.43-7.03(m,4H),6.06(s,1H),4.01(s,2H),3.61-3.48(m,1H),1.76-1.47(m,5H),1.31-1.05(m,14H).

[0751] Example 42. N-(4-(tert-butyl)phenyl)-2-chloro-N-(1-(6-cyanopyridin-3-yl)-2-(cyclohexylamino)- Synthesis of 2-(2-(2-methyl)-2-oxoethyl)acetamide (INSCoV-539A)

[0752] INSCoV-539A was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was triturated with ACN for 30 min at 25 ° C. Compound (2R)-2-(4-tert-butyl-N-(2-chloroacetyl)anilino)-2-(6-cyano-3-pyridinyl)-N-cyclohexyl-acetamide (350.65 mg, 737.50 μmol, 55.03% yield, 98.222% purity) was obtained as a white solid. 1 HNMR, LCMS and HPLC confirmed the correct structure.

[0753] LCMS: retention time: 1.102 min, (M+H) = 467.2. HPLC: retention time: 2.939 min. 1 H NMR (400MHz, chloroform-d): δ = 8.55 (s, 1H), 7.65 (br d,J=8.1Hz,1H),7.48(s,1H),7.37-7.29(m,2H),7.27(s,1H),7.09-6.79(m,1H) ,6.22-6.14(m,1H),6.09(s,1H),3.87-3.84(m,2H),3.84-3.77(m,1H),2.00(br d,J=10.8Hz,1H),1.88(br dd,J=1.7,10.8Hz,1H),1.80-1.66(m,2H),1.61(s,5H),1.46-1.33(m,2H),1.29(s,9H),1.19(br d,J=12.2Hz,3H).

[0754] Example 43. Synthesis of INSCoV-549

[0755] Plan 23

[0756]

[0757] INSCoV-549 was synthesized according to the general procedure for the INSCoV series. Purification B: The mixture was poured into water (20 ml), and DCM (20 ml) was added, the organic layer was washed with brine (20 ml), dried over Na2SO4 and concentrated in vacuo. The crude product was ground with MTBE (5 ml), filtered, and the filter cake was ground with EtOH (2 ml), filtered, and the filter cake was dried in vacuo. INSCoV-549 (154.05 mg, 309.88 μmol, 29.28% yield, 96.763% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0758] HPLC: retention time: 2.462 min. LCMS: retention time: 0.937 min, (M+H)=481.2. 1 H NMR (400MHz, DMSO-d6): δ = 11.46 (br s, 1H), 8.07 (d, J = 4.0Hz, 1H), 7.81 (br d, J = 78.0Hz, 1H), 7.55 (br d,J=8.0Hz,1H),7.24-6.77(m,4H),6.30(s,1H),4.01-3.85(m,2H),3.66-3.52(m,1H),1.82-1.46(m,5H),1.34-0.94(m,14H).

[0759] Example 44. N-(4-(tert-butyl)-2-(3-morpholinoprop-1-yn-1-yl)phenyl)-2-chloro-N-(2-(cyclopentyl)-1-ol Synthesis of (1-(hexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)acetamide (INSCoV-553)

[0760] INSCoV-553 was synthesized according to the general procedure for the INSCoV series. Purification B: The residue was triturated with MeCN (3 mL) and then filtered. The filter cake was washed with MeCN (2 mL × 2) and dried in vacuo. INSCoV-553 (72.6 mg, 123 μmol, 33.6% yield, 96.0% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS and HPLC.

[0761] LCMS: retention time: 1.025 min, [M+H + ] = 565.3. HPLC: retention time: 2.741 min. 1H NMR (400MHz, DMSO-d6): δ = 8.31 (d, J = 2.1Hz, 1H), 8.27 (dd, J = 1.2, 4.8Hz, 1H), 8.08 (d, J = 7.6Hz, 1H), 7.85 (d, J =8.4Hz,1H),7.45-7.40(m,1H),7.27(d,J=8.1Hz,1H),7.14(d,J=2.4Hz,1H),7.03(dd,J=4.8,8.0Hz,1H),6.02 (s,1H),4.02-3.92(m,1H),3.90-3.81(m,1H),3.63(t,J=4.8Hz,4H),3.60-3.54(m,1H),3.50(d,J=2.8Hz,2H) ,3.31(s,2H),1.85-1.73(m,1H),1.73-1.65(m,1H),1.63-1.44(m,3H),1.28-1.12(m,13H),1.11-0.86(m,2H).

[0762] Example 45. Synthesis of INSCoV-557A

[0763] Plan 24

[0764]

[0765] Step 1: To a solution of compound 1 (3 g, 18.50 mmol, 1 eq) and compound 2 (1.91 g, 22.20 mmol, 1.2 eq) in DCE (120 mL) was added NaHCO (3.11 g, 37.00 mmol, 1.44 mL, 2 eq), Cu(OAc) (3.36 g, 18.50 mmol, 1 eq), and 2-(2-pyridyl)pyridine (2.89 g, 18.50 mmol, 1 eq). The reaction was stirred at 25° C. under O (15 PSI) for 4 days. The mixture was then heated to 60° C. under O (15 PSI) for 24 h. LCMS showed that compound 1 remained, and a new peak with the desired mass was detected (Rt=0.917 min). TLC (PE:EA=5:1) showed that compound 1 (Rf=0.2) remained and two new spots (Rf=0.02, Rf=0.7) were formed. The mixture was diluted with water (50 mL) and extracted with DCM (70 mL×3). The organic layer was dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 80g The residue was purified by silica flash column, 0-30% ethyl acetate / petroleum ether gradient at 100 mL / min and concentrated in vacuo to obtain compound 3 (1.6 g, 7.91 mmol, 42.77% yield) as a yellow solid, which was confirmed by HNMR.

[0766] LCMS: retention time: 0.972 min, (M+H)=203.1. 1 H NMR (400MHz, DMSO-d6)δ=8.13-8.07(m,2H),7.78-7.72(m,1H),7.43-7.37(m,1H) ,6.98(d,J=3.2Hz,1H),3.62-3.56(m,1H),1.18-1.07(m,2H),1.07-0.95(m,2H).

[0767] Step 2: Pd / C (0.1 g, 4.95 mmol, 1 eq) was added to a solution of compound 3 (1 g, 4.95 mmol, 1 eq) in MeOH (20 mL). The reaction mixture was stirred for 24 h at 25 ° C under H2 (50 PSI). LCMS showed that compound 3 was completely consumed, and a new peak with the desired mass was detected (Rf = 0.903 min). TLC (PE: EA = 5: 1) showed that compound 3 (Rf = 0.5) was completely consumed, and a new spot (Rf = 0.3) was formed. The reaction mixtures were combined for post-processing. The reaction mixture was adjusted to pH = 10 by NH3·H2O, filtered through a celite pad, and washed with MeOH (40 mL × 2). The filtrate was concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica flash column (0-80% ethyl acetate / petroleum ether gradient at 80 mL / min). The combined organic phases were concentrated in vacuo. Compound 4 (0.9 g, 5.23 mmol, 105.67% yield) was obtained as a red oil, which was confirmed by H NMR.

[0768] LCMS: retention time: 0.903 min, (M+H)=173.2. 1 H NMR (400MHz, DMSO-d6) δ = 7.03 (d, J = 3.3Hz, 1H), 6.88-6.80 (m, 1H), 6.73 (d, J = 8.0Hz, 1H), 6.45 (d, J = 2.8Hz,1H),6.20–6.14(m,1H),5.17(s,2H),3.31-3.27(m,1H),1.04-0.95(m,2H),0.90-0.82(m,2H).

[0769] Step 3: INSCoV-557A is synthesized according to the general procedure for the INSCoV series. Purification A: The reaction is concentrated in vacuo. The crude product is purified by reverse phase HPLC (0.1% FA) and concentrated in vacuo to remove MeCN. The aqueous phase is lyophilized to produce a crude product. The residue is dissolved in DMF (2 mL) and purified by preparative HPLC (column: Waters Xbridge 150 × 25 mm × 5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 35%-65%, 8 min) and diluted with water (30 mL), and the liquid is lyophilized to produce the product. INSCoV-557-A (7.16 mg, 13.79 μmol, 1.49% yield, 96.703% purity) was obtained as a yellow solid, which was confirmed by LCMS, HPLC, SFC, HNMR and FNMR.

[0770] LCMS: Retention time: 0.913 min, (M+H) = 502.0. HPLC: Retention time: 2.196 min. SFC: Retention time: 3.036 min, 3.281 min. 1 H NMR (400MHz, DMSO-d6) δ = 8.88 (s, 1H), 8.72 (s, 1H), 8.52 (s, 1H), 8.41 (s, 2H), 8.25-8.11 (m, 1H), 7.53 (d,J=7.6Hz,1H),7.48(d,J=8.4Hz,1H),7.42(d,J=3.2Hz,1H),7.24(d,J=3.2Hz,1H),7.22-7.14(m,1H ),7.04-6.98(m,1H),6.86-6.77(m,1H),6.24(d,J=3.2Hz,1H),6.10(s,1H),5.88-5.78(m,1H),5.82(s ,1H),4.07-3.67(m,5H),3.49-3.42(m,1H),2.08-1.47(m,11H),1.39-1.22(m,2H),1.08-0.77(m,7H). 19 F NMR (377MHz, DMSO-d6) δ=-92.05--95.13(m,1F),-96.91--99.42(m,1F).

[0771] Example 46. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(3-fluorobenzene Synthesis of (ethyl)acetamide (INSCoV-558)

[0772] INSCoV-558 was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was triturated by ACN (10 mL) at 4 ° C to give the pure product 2-[(2-chloroacetyl)-[2-(3-fluorophenyl)ethyl]amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (105.75 mg, 236.21 μmol, 16.44% yield, 96.477% purity) obtained as a white solid. The compound 2-[(2-chloroacetyl)-[2-(3-fluorophenyl)ethyl]amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (105.75 mg, 236.21 μmol, 16.44% yield, 96.477% purity) was obtained as a white solid. LCMS, HPLC, 1 The correct structure was verified by HNMR and FNMR.

[0773] LCMS: retention time: 0.970 min, (M+H)=432.1. HPLC: retention time: 2.482 min 。 1 H NMR (400MHz, chloroform-d): δ = 8.71-8.61 (m, 2H), 7.89 (br d, J = 7.7Hz, 1H), 7.38 (dd, J = 4.8, 7.9Hz, 1H), 7.26-7.16 (m, 1H), 6.92 (br t, J = 8.1Hz, 1H), 6.82 (br d,J=7.7Hz,1H),6.73(br d,J=9.5Hz,1H),6.02(br d,J=6.2Hz,1H),5.80(s,1H),3.99(s,2H),3.90-3.77(m,1H),3.60(br t,J=7.6Hz,2H),2.88-2.75(m,1H),2.58-2.45(m,1H),1.99-1.88(m,2H),1 .74-1.66(m,2H),1.61-1.56(m,1H),1.44-1.31(m,2H),1.23-1.08(m,3H).

[0774] Example 47. 2-Chloro-N-(2-((1,1-dioxotetrahydro-2H-thiopyran-4-yl)amino)-2-oxo-1-(pyrimidin- Synthesis of (5-pyridin-5-yl)ethyl)-N-(3-fluorophenethyl)acetamide (INSCoV-558A)

[0775] INSCoV-558A was synthesized according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 Mm NH4HCO3)-ACN]; B%: 24%-54%, 10 min). 2-[(2-chloroacetyl)-[2-(3-fluorophenyl)ethyl]amino]-N-(1,1-dioxothianilan-4-yl)-2-pyrimidin-5-yl-acetamide (5.7 mg, 10.81 μmol, 1.77% yield, 91.597% purity) was obtained as a white solid, which was detected by LCMS, HPLC and HNMR.

[0776] LCMS: retention time: 0.861 min, [M+H+] = 483.2. HPLC: retention time: 1.896 min. 1 H NMR: (400MHz, DMSO-d6): δ = 9.14 (s, 1H), 8.74 (s, 2H), 8.05 (br d,J=7.8Hz,1H),7.42-7.20(m,1H),7.15-6.88(m,3H),5.68(s,1H),4.60-4.50(m,2H),4.14-3.97(m,1H),3.58(br t,J=8.3Hz,2H),3.27-3.20(m,2H),3.11-2.98(m,3H),2.92-2.75(m,1H),2.64-2.59(m,1H),2.03-1.88(m,4H).

[0777] Example 48. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(4-fluorobenzene Synthesis of (ethyl)acetamide (INSCoV-559)

[0778] INSCoV-559 was synthesized according to the general procedure for the INSCoV series. Purification A: The residue was purified by preparative HPLC (column: Phenomenex Synergi C18 150×25 mm×10 μm; mobile phase: [water (0.1% TFA)-ACN]; B%: 25%-55%, 10 min) followed by LCMS verification, and 73% of the desired mass was detected. The crude product was repurified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 39%-69%, 9 min) followed by LCMS verification, and 77% of the desired mass was detected. The crude product was repurified by preparative HPLC (column: Phenomenex Gemini-NX C18 75×30 mm×3 um; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 31%-61%, 11.5 min), followed by LCMS verification, and 100% of the desired mass was detected. Compound 2-[(2-chloroacetyl)-[2-(4-fluorophenyl)ethyl]amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (15.23 mg, 35.26 μmol, 4.91e-1% yield) was obtained as a white solid. 1 The structure was verified by HNMR.

[0779] LCMS: retention time: 0.978 min, (M+H) = 432.1. HPLC: retention time: 2.498 min. 1 H NMR (400MHz, chloroform-d): δ=8.71-8.61(m,2H),7.93-7.83(m,1H),7.41-7.33(m,1 H),7.02-6.93(m,4H),6.02-5.93(m,1H),5.81-5.75(m,1H),4.01-3.93(m,2H ),3.87-3.78(m,1H),3.64-3.52(m,2H),2.86-2.71(m,1H),2.57-2.42(m,1H) ,2.01-1.86(m,2H),1.78-1.64(m,3H),1.46-1.30(m,3H),1.23-1.10(m,4H).

[0780] Example 49. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(2-methoxy Synthesis of (4-(2-aminophenylethyl)acetamide (INSCoV-560A)

[0781] INSCoV-560A was synthesized according to the general procedure for the INSCoV series. The residue was ground from MeOH (10 mL) to produce the pure product 2-[(2-chloroacetyl)-[2-(2-methoxyphenyl)ethyl]amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (800 mg, 1.80 mmol, 27.25% yield, 100% purity) as a white solid. Compound 2-[(2-chloroacetyl)-[2-(2-methoxyphenyl)ethyl]amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (800 mg, 1.80 mmol, 27.25% yield, 100% purity) as a white solid was obtained. LCMS, HPLC and 1 HNMR confirmed the correct compound.

[0782] LCMS: retention time: 0.980 min, (M+H) = 444.2. HPLC: retention time: 2.522 min. 1 H NMR (400MHz, chloroform-d): δ=8.70-8.62(m,2H),7.92-7.86(m,1H),7.39-7.33(m ,1H),7.24-7.17(m,1H),6.83(d,J=5.1Hz,2H),6.12-6.04(m,1H),5.92-5. 89(m,1H),4.28(d,J=9.0Hz,2H),3.84(s,4H),3.66-3.55(m,1H),3.55-3.4 4(m,1H),2.84-2.73(m,1H),2.37-2.27(m,1H),1.98-1.87(m,2H),1.67(br d,J=3.5Hz,1H),1.69(br s,1H),1.63-1.55(m,1H),1.44-1.29(m,2H),1.23-1.08(m,3H).

[0783] Example 50. N-(4-(tert-butyl)phenyl)-2-chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridine-3- Synthesis of (4-amino)ethyl)acrylamide (INSCoV-570)

[0784] INSCoV-570 was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was triturated with EtOAc (1 mL) and petroleum ether (15 mL) at 20 ° C for 60 min. INSCoV-570 (142 mg, 297 μmol, 44.36% yield, 95.204% purity) was obtained as a white solid. The product was purified by LCMS, HPLC and 1 HNMR to verify.

[0785] LCMS: retention time: 1.070 min, [M] = 456.2; retention time: 3.029 min, [M] = 456.2. HPLC: retention time: 2.939 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.34-8.24 (m, 2H), 8.01 (dd, J = 7.6, 15.2Hz, 1H), 7.69-6.98 (m, 5H), 5.98 (d, J=17.6Hz,1H),4.18(qd,J=6.6,13.6Hz,1H),3.63-3.48(m,1H),1.78-1.41(m,8H),1.32-0.87(m,15H).

[0786] Example 51. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(3-(trifluoromethyl)- Synthesis of (methyl)benzyl)acetamide (INSCoV-574)

[0787] INSCoV-574 was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was purified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 36%-69%, 9 min). INSCoV-574 (153.85 mg, 327.55 μmol, 28.69% yield, 99.62% purity) was obtained as a white solid, which was detected by LCMS, HPLC, HNMR and FNMR.

[0788] LCMS: retention time: 0.974 min, [M+H + ]=468.2;Retention time;2.449min,[M+H + ]=468.2.HPLC: retention time: 2.715 min. 1 H NMR (400MHz, DMSO-d6, T=80): δ=8.50-8.34(m,2H),7.93(br d,J=6.8Hz,1H),7.66(br d,J=8.0Hz,1H),7.54-7.10(m,5H),6.08-5.83(m,1H),4.96(d,J=17.2Hz,1H),4.67(br d,J=17.0Hz,1H),4.50-4.25(m,2H),3.66-3.51(m,1H),1.78-1.50(m,5H),1.31-1.09(m,5H). 19 F NMR (400MHz, DMSO-d6): -61.52 (s, 3F).

[0789] Example 52. 2-Chloro-N-(2-(cyclohexylamino)-2-oxo-1-(pyridin-3-yl)ethyl)-N-(2-(trifluoro Synthesis of (methyl)benzyl)acetamide (INSCoV-574A)

[0790] INSCoV-574A was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was purified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 38%-68%, 9 min). INSCoV-574A (254.01 mg, 533.37 μmol, 46.71% yield, 98.252% purity) was obtained as a yellow solid, which was detected by LCMS, HPLC, HNMR and FNMR.

[0791] LCMS: retention time: 0.974 min, [M+H + ]=468.2.HPLC: retention time: 2.705 min. 1 H NMR (400MHz, DMSO-d6, T=80): δ=8.64-8.26(m,2H),8.21-7.89(m,1H),7.79-7.04(m,6H),6.24-5.91(m,1H),5.16 -4.67(m,2H),4.62-4.23(m,2H),3.76-3.38(m,1H),1.88-1.43(m,5H),1.33-1.08(m,5H). 19 F NMR: (400MHz, DMSO-d6): -60.52 (s, 3F).

[0792] Example 53. Synthesis of INSCoV-575

[0793] Plan 25

[0794]

[0795] INSCoV-575 was synthesized according to the general procedure for the INSCoV series. Purification B: The residue was poured into ACN (5 mL) and stirred for 5 min, and the crystalline solid was collected by suction filtration. The filtrate was adjusted to pH = 7-8 and discarded. INSCoV-575 (365.95 mg, 785.73 μmol, 63.33% yield, 97.47% purity) was obtained as a white solid, which was detected by LCMS, HPLC and HNMR.

[0796] LCMS: retention time: 1.005 min, [M+H + ]=454.2.HPLC: retention time: 2.777 min. 1H NMR (400MHz, DMSO-d6): δ = 8.39-8.22 (m, 2H), 8.11 (br d,J=6.7Hz,1H),8.02-7.55(m,1H),7.47-7.29(m,1H),7.22-7.00(m,2H),7. 00-6.46(m,1H),6.07(s,1H),4.11-3.82(m,2H),3.68-3.51(m,1H),2.33(br s,3H),2.19-1.85(m,3H),1.83-1.46(m,5H),1.38-0.86(m,5H).

[0797] Example 54. Synthesis of INSCoV-576

[0798] Plan 26

[0799]

[0800] Step 1: At 25 ° C, under N2, to a mixture of 2-hydroxy-4-nitro-benzaldehyde (2g, 11.97mmol, 1eq) and diethyl 2-bromomalonate (2.86g, 11.97mmol, 2.04mL, 1eq) in 2-butanone (50mL) was added K2CO3 (4.96g, 35.90mmol, 3eq) at once, then heated to 90 ° C and stirred for 16 hours. LCMS shows that the reaction is complete. The mixture is poured into ice water (100mL) and stirred for 5min. The aqueous phase is extracted with ethyl acetate (100mL×3), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue is purified by flash silica gel chromatography ( 40g The product was purified by silica flash column with a gradient of 0-100% ethyl acetate / petroleum ether at 60 mL / min. TLC (PE:EA=2:1, RF=0.6) was performed to obtain ethyl 6-nitrobenzofuran-2-carboxylate (2.7 g, 10.91 mmol, 91.13% yield, 95% purity) as a light yellow solid, which was analyzed by LCMS and 1 HNMR was used for detection.

[0801] LCMS: retention time: 0.909 min, [M+H+] = 236.2: retention time: 1.207 min, [M+H+] = 236.2. 1HNMR (400 MHz, CHLOROFORM-d): δ = 8.53-8.47 (m, 1H), 8.25 (dd, J = 2.0, 8.8 Hz, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.61 (d, J = 1.0 Hz, 1H), 4.49 (q, J = 7.2 Hz, 2H), 1.58 (s, 3H).

[0802] Step 2: at 25 ° C, under N2, to a mixture of 6-nitrobenzofuran-2-carboxylic acid ethyl ester (2g, 8.50mmol, 1eq) in EtOH (20mL) was added KOH (715.65mg, 12.76mmol, 1.5eq) at one time, then heated to 70 ° C and stirred for 1 hour. LCMS and HPLC showed that the reaction was complete. The solvent was evaporated under reduced pressure, and the residue was dissolved in water (10mL) and acidified to pH 4 with concentrated hydrochloric acid. The resulting precipitate was collected by filtration, washed with water and dried. 6-nitrobenzofuran-2-carboxylic acid (1.6g, crude product) was obtained as a light yellow solid. 1 HNMR was used for detection.

[0803] LCMS: retention time: 0.200 min, [M+H + ] = 208.1. HPLC: retention time: 0.249 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.67 (s, 1H), 8.37-8.15 (m, 1H), 8.03 (br d, J = 8.7Hz, 1H), 7.83 (s, 1H).

[0804] Step 3: At room temperature, under nitrogen, to a mixture of 6-nitrobenzofuran-2-carboxylic acid (500 mg, 2.41 mmol, 1 eq) in DMSO (5 mL) was added AgCO (332.80 mg, 1.21 mmol, 54.74 μL, 0.5 eq) and AcOH (14.50 mg, 241.38 μmol, 13.81 μL, 0.1 eq). The reaction mixture was stirred for 3 hours at 120 ° C. LC-MS showed that the reaction was complete and the desired product was obtained. The mixture was filtered. The mixture was poured into ice water (20 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 3), dried over anhydrous NaSO, filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 40g Silica flash column, 0-100% ethyl acetate / petroleum ether gradient eluent, at 60 mL / min) was used for purification. TLC (PE / EA=3:1, RF=0.5) was used to obtain 6-nitrobenzofuran (390 mg, 2.22 mmol, 92.11% yield, 93% purity) as a white solid. 1 HNMR was used for detection.

[0805] LCMS: retention time: 0.915 min, [M+H + ]=164.1. 1 H NMR (400 MHz, CHLOROFORM-d): δ = 8.44 (s, 1H), 8.20 (dd, J = 2.0, 8.6 Hz, 1H), 7.90 (d, J = 2.4 Hz, 1H), 7.71 (d, J = 8.6 Hz, 1H), 6.92 (dd, J = 0.9, 2.1 Hz, 1H).

[0806] Step 4: 6-nitrobenzofuran (350 mg, 2.15 mmol, 1 eq) was dissolved in a mixed solvent of MeOH (20 mL) and THF (20 mL), and Pd / C (350 mg, 10% purity) was then added thereto. At 25 ° C, under H2 (50 psi), the reaction mixture was stirred for 3 hours. LC-MS showed that the reaction was complete and the desired product was obtained. After filtration and concentration under reduced pressure, 2,3-dihydrobenzofuran-6-amine (270 mg, crude product) was obtained as a brown oil.

[0807] LCMS: retention time: 0.777 min, [M+H + ]=136.2.

[0808] Step 5: To a mixture of 2,3-dihydrobenzofuran-6-amine (250 mg, 1.85 mmol, 1 eq) in dioxane (10 mL) was added DDQ (461.86 mg, 2.03 mmol, 1.1 eq) in one portion at 25°C under N2, followed by heating to 90°C and stirring for 2 hours. LC-MS showed 39% of 2,3-dihydrobenzofuran-6-amine remaining and 14% of the desired compound. The mixture was filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica flash column (0-100% ethyl acetate / petroleum ether gradient at 50 mL / min). TLC (PE / EA=3 / 1, RF=0.5) was used to obtain benzofuran-6-amine (120 mg, 865.21 μmol, 46.78% yield, 96% purity) as a brown oil, which was detected by LCMS.

[0809] LCMS: retention time: 0.703 min, [M+H + ]=134.2.

[0810] Step 6: INSCoV-576 was synthesized according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 36%-66%, 8 min). INSCoV-576 2-[benzofuran-6-yl-(2-chloroacetyl)amino]-N-cyclohexyl-2-(3-pyridyl)acetamide (80.51 mg, 184.23 μmol, 30.66% yield, 97.46% purity) was obtained as a white solid, which was detected by LCMS, HPLC and HNMR.

[0811] LCMS: retention time: 0.920 min, [M+H + ] = 426.2. HPLC: retention time: 2.452 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.44-8.23 (m, 2H), 7.92 (d, J = 2.1Hz, 1H), 7.83-7.54 (m, 1H), 7.82-7.51 (m, 1H), 7.51-7. 37(m,2H),7.21-7.00(m,2H),6.86(dd,J=0.9,2.2Hz,1H),6.14(s,1H),4.07-3.78(m,2H),3.71-3.49(m,1H),1.94 -1.46(m,5H),1.41-1.00(m,5H).

[0812] Example 55. Synthesis of INSCoV-600A

[0813] Plan 27

[0814]

[0815] Step 1: At 70 ° C, under N2, a mixture of compound 1 (5.0 g, 49.43 mmol, 1 eq) and compound 2 (46.05 g, 621.64 mmol, 50 mL, 12.58 eq) was stirred for 12 h. TLC (PE / EA=3 / 1) indicated that compound 1 (Rf=0.0) was consumed and a small spot (Rf=0.1) was formed. The mixture was concentrated in vacuo. The reaction was used for the next step without purification. Compound 3 (7.0 g, crude product) was obtained as a white solid.

[0816] 1 H NMR (400 MHz, chloroform-d): δ = 8.14 (s, 1H), 5.79-5.17 (m, 1H), 4.19-4.05 (m, 1H), 4.03-3.92 (m, 2H), 3.52-2.46 (m, 2H), 2.02-1.89 (m, 2H), 1.56-1.41 (m, 2H).

[0817] Step 2: To a solution of compound 3 (1.0 g, 7.74 mmol, 1 eq) and TEA (783.46 mg, 7.74 mmol, 1.08 mL, 1.0 eq) in DCM (10 mL) was added PPh (2.23 g, 8.52 mmol, 1.1 eq). The mixture was stirred at 45 ° C under N2 for 12 h. TLC (PE / EA=3 / 1) indicated that compound 3 (Rf=0.1) was consumed and a small spot (Rf=0.6) was formed. The resulting mixture was evaporated in vacuo (<20 ° C). At 20 ° C, under N2, the residue was suspended in Et2O (100 ml) for 12 h. The solid was filtered, washed with Et2O (100 mL×2), and the filtrate was concentrated in vacuo (<20 ° C). The combined organic phase was concentrated in vacuo (<20 ° C). The reaction was used for the next step without purification. Compound 4 (0.8 g, 7.20 mmol, 92.97% yield) was obtained as a brown oil. 1 H NMR to confirm.

[0818] 1 H NMR (400 MHz, chloroform-d): δ = 3.90-3.71 (m, 3H), 3.58-3.45 (m, 2H), 1.99-1.86 (m, 2H), 1.83-1.73 (m, 2H).

[0819] Step 3: INSCoV-600A was synthesized according to the general procedure for the INSCoV series. Purification B: MTBE (20 mL) was added to the residue, the suspension was filtered and washed with MTBE (10 mL × 3) to give a crude product. The residue was ground with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-600A (147.51 mg, 305.05 μmol, 32.97% yield, 94.278% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0820] LCMS: Retention time: 0.737 min, (M+H) = 456.1. HPLC: Retention time: 1.460 min. SFC: Retention time: 1.469 min, 1.796 min. 1 H NMR (400MHz, DMSO-d6): δ=9.09-9.00(m,2H),8.45(s,1H),8.39(d,J=7.6Hz,1H),7.71(s,1H),7.63(d,J=8.8Hz,2H),7.54-7.42(m,2H),7. 40-7.34(m,1H),6.07(s,1H),4.11-4.05(m,2H),3.87-3.74(m,3H),3 .34(s,1H),3.32-3.26(m,1H),1.75-1.61(m,2H),1.48-1.21(m,2H).

[0821] Example 56. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyridin-3-yl)-2-(tetramethyl-1-oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyridin-3-yl)phenyl)-2-(tetramethyl-1-oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyridin-3 Synthesis of (2H-pyran-4-yl)amino)ethyl)acetamide (INSCoV-600A(1))

[0822] INSCoV-600A (1) was synthesized according to the general procedure for the INSCoV series. Purification B: The residue was triturated with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-600A (1) (123.82 mg, 267.11 μmol, 28.61% yield, 98.136% purity) was obtained as a white solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0823] LCMS: Retention time: 0.693 min, (M+H) = 455.1. HPLC: Retention time: 1.251 min. SFC: Retention time: 1.452 min, 1.678 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.43 (s, 1H), 8.36-8.24 (m, 3H), 7.69 (s, 1H), 7 .66-7.44(m,3H),7.41-7.35(m,1H),7.18-7.12(m,1H),6.08(s,1H),4.10 -3.94(m,2H),3.87-3.70(m,3H),3.40-3.35(m,1H),3.32-3.28(m,1H),1. 77-1.71(m,1H),1.66-1.60(m,1H),1.52-1.37(m,1H),1.31-1.16(m,1H).

[0824] Example 57. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyrimidin-5-yl)-2-(tetramethyl- Synthesis of (2H-pyran-4-yl)amino)ethyl)acetamide (INSCoV-600A(2))

[0825] INSCoV-600A (2) was synthesized according to the general procedure for the INSCoV series. Purification B: MTBE (20 mL) was added to the reaction mixture, filtered and washed with MTBE (10 mL × 3) to give a crude product. The residue was triturated with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-600A (2) (202.45 mg, 435.58 μmol, 47.09% yield, 98.087% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0826] LCMS: retention time: 0.743 min, (M+H) = 456.1. HPLC: retention time: 1.455 min. SFC: retention time: 0.515 min, 0.945 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.97 (s, 1H), 8.53-8.44 (m, 3H), 8.34 (d, J = 7.2Hz, 1H), 7.72 (s, 1H), 7.68-7.62 (m, 2H), 7.44 (d, J = 3.6Hz, 2H), 6.10 (s, 1H),4.16-3.99(m,2H),3.88-3.68(m,3H),3.36(d,J=2.0Hz,1H),3.30(d ,J=2.0Hz,1H),1.81-1.59(m,2H),1.52-1.38(m,1H),1.34-1.20(m,1H).

[0827] Example 58. 2-Chloro-N-(4-(oxazol-5-yl)phenyl)-N-(2-oxo-1-(pyrazin-2-yl)-2-(tetra Synthesis of (2H-pyran-4-yl)amino)ethyl)acetamide (INSCoV-600A(3))

[0828] INSCoV-600A (3) was synthesized according to the general procedure for the INSCoV series. Purification A: The residue was triturated with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-600A (3) (177.52 mg, 375.02 μmol, 40.54% yield, 96.309% purity) was obtained as an off-white solid, which was confirmed by HNMR, LCMS, SFC and HPLC.

[0829] LCMS: retention time: 0.746 min, (M+H)=456.1. HPLC: retention time: 1.502 min. SFC: retention time: 1.720 min, 2.182 min. 1H NMR (400MHz, DMSO-d6): δ = 8.49 (s, 2H), 8.46-8.40 (m, 2H), 8.32 (d, J = 7.6Hz, 1H), 7.70 (s, 1H), 7.61 (d, J = 8.4Hz, 2H), 7.50-7.44 (m, 1H), 6 .23(s,1H),4.21-4.00(m,2H),3.87-3.71(m,3H),3.35(d,J=2.8Hz,1H),3.30(d,J=3.2Hz,1H),1.66(d,J=11.2Hz,2H),1.45-1.24(m,2H).

[0830] Example 59. Synthesis of INSCoV-600B

[0831] Plan 28

[0832]

[0833] Step 1: A mixture of 3,3-difluorocyclopentylamine hydrochloride (1 g, 6.35 mmol, 1 eq) and TEA (1.28 g, 12.69 mmol, 1.77 mL, 2 eq) in ethyl formate (2.35 g, 31.73 mmol, 2.55 mL, 5 eq) was stirred at 80 ° C for 16 h. TLC (EA: EtOH = 3: 1) showed that the starting material (Rf = 0.3) was completely consumed. And a new spot (Rf = 0.7) was detected. The mixture was concentrated under reduced pressure. The crude compound was used in the next step without further purification. Compound 2 (940 mg, 6.30 mmol, 99.33% yield) was obtained as a yellow oil. 1 HNMR to verify.

[0834] 1 H NMR: (400MHz, CDCl3): δ = 8.17-8.06 (m, 1H), 4.59-4.40 (m, 1H), 2.56-2.47 (m, 2H), 1.57 (br t, J = 7.2Hz, 2H), 1.24 (br t, J = 7.2Hz, 2H).

[0835] Step 2: at -10 ° C, under N2, to a mixture of compound 2 (500 mg, 3.35 mmol, 1 eq) and DIEA (2.17 g, 16.7 mmol, 2.92 mL, 5 eq) in DCM (100 mL) was added POCl3 (616.86 mg, 4.02 mmol, 373.86 μ L, 1.2 eq), then heated to 20 ° C and stirred for 2 hours. TLC (plate 1, PE: EA = 1: 1) showed that compound 2 (Rf = 0.1) was consumed, and a new spot (Rf = 0.8) was observed. At 0 ° C, the mixture was slowly poured into saturated NaHCO3 (500 mL), then extracted with DCM (300 mL × 2), dried over Na2SO4. At 25 ° C, the mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 40g The mixture was purified by silica flash column (0-50% ethyl acetate / petroleum ether gradient at 50 mL / min). The mixture was concentrated under reduced pressure at 25°C. TLC (plate 2, PE:EA=1:1, Rf=0.1) afforded compound 3 (200 mg, 1.53 mmol, 45.5% yield) as a colorless oil.

[0836] Step 3: INSCoV-600B was synthesized according to the general procedure for the INSCoV series. Purification B and Purification A: The residue was triturated with MeCN (5 mL) at 25°C for 30 min and filtered. The filter cake was repurified by preparative HPLC (column: Welch MLtimate XB-CN 250×50×10 um; mobile phase: [hexane-IPA]; B%: 25%-65%, 15 min) and concentrated in vacuo to produce the desired compound. The filtrate was concentrated in vacuo to produce a residue. The residue was repurified by preparative HPLC (column: Welch MLtimate XB-CN 250×50×10 um; mobile phase: [hexane-IPA]; B%: 25%-65%, 15 min), but this failed to produce the desired compound. 2-(N-(2-Chloroacetyl)-4-oxazol-5-yl-anilino)-N-(3,3-difluorocyclopentyl)-2-pyrimidin-5-yl-acetamide (73.31 mg, 148 μmol, 15.8% yield, 96.04% purity) was obtained as a yellow solid, which was verified by HNMR, FNMR, LCMS and HPLC.

[0837] LCMS: retention time: 0.812 min, (M+H) = 442.2. HPLC: retention time: 1.560 min. 1H NMR: (400MHz, DMSO-d6): δ = 8.98 (d, J = 2.0Hz, 1H), 8.60-8.51 (m, 1H), 8.51-8.48 (m, 2H), 8.46 (s, 1H), 7.75-7.70 (m, 1H), 7.66 (br d,J=8.4Hz,2H),7.53-7.31(m,2H),6.07(d,J=2.4Hz,1H),4.31-4.18(m,1H),4.13-3. 99(m,2H),3.82-3.72(m,1H),2.46-2.35(m,1H),2.25-1.80(m,4H),1.75-1.63(m,1H). 19 F NMR (377MHz, DMSO-d6): δ = -88.33 (s, 1F), -88.47--88.63 (m, 1F).

[0838] Example 60. Synthesis of INSCoV-600C

[0839] Plan 29

[0840]

[0841] Step 1: A mixture of tetrahydrofuran-3-amine (2 g, 22.96 mmol, 1 eq) in ethyl formate (8.50 g, 114.78 mmol, 9.23 mL, 5 eq) was stirred at 80 ° C for 16 h. TLC (EA: EtOH = 3: 1) showed that the starting material (Rf = 0.3) was completely consumed. And a new spot (Rf = 0.7) was detected. The mixture was concentrated under reduced pressure. The crude compound was used in the next step without further purification. N-tetrahydrofuran-3-ylformamide (2.5 g, 21.71 mmol, 94.59% yield) was obtained as a yellow oil. 1 H NMR to verify.

[0842] 1 H NMR (400MHz, CDCl3): δ = 8.10 (s, 1H), 6.53-6.25 (m, 1H), 4.61-4.53 (m, 1H), 3.83-3.76 (m, 2H), 3.74 -3.62 (m, 2H), 2.42-2.12 (m, 2H).

[0843] Step 2: At -10 ° C, under N2, to a mixture of N-tetrahydrofuran-3-ylformamide (1.5 g, 13.03 mmol, 1 eq) and DIEA (8.42 g, 65.1 mmol, 11.4 mL, 5 eq) in DCM (100 mL) was added POCl3 (2.40 g, 15.63 mmol, 1.45 mL, 1.2 eq) at once, then heated to 20 ° C and stirred for 2 hours. TLC (PE: EA = 1: 1) showed that N-tetrahydrofuran-3-ylformamide (Rf = 0.1) was consumed, and a new spot (Rf = 0.8) was observed. At 0 ° C, the mixture was slowly poured into saturated NaHCO3 (500 mL), then extracted with DCM (300 mL × 2), dried over Na2SO4. At 25 ° C, the mixture was concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 40g The product was purified by silica flash column with a gradient of 0-50% ethyl acetate / petroleum ether at 50 mL / min. The mixture was concentrated under reduced pressure at 25° C. Compound 3 (500 mg, 5.15 mmol, 39.52% yield) was obtained as a colorless oil and used directly in the next step.

[0844] Step 3: INSCoV-600C was synthesized according to the general procedure for the INSCoV series. Purification B: The crude product was triturated with MeCN (5 mL) at 25° C. for 30 min. INSCoV-600C (198.49 mg, 438.65 μmol, 35.1% yield, 97.65% purity) was obtained as an off-white solid, which was verified by HNMR, LCMS, and HPLC.

[0845] LCMS: retention time: 0.812 min, (M+H) = 442.2. HPLC: retention time: 1.560 min. 1 H NMR (400MHz, DMSO-d6): δ = 9.02 (s, 1H), 8.66 (dd, J = 4.0, 6.4Hz, 1H), 8.59-8.44 (m, 3H), 7.77 (s, 1H), 7.66 (br s, 1H), 7.70 (br d,J=8.4Hz,1H),7.61-7.33(m,2H),6.13(s,1H),4.34(tdd,J=3.6,6.4,10.0Hz,1H),4.18-3.9 9(m,2H),3.87-3.63(m,3H),3.56(dd,J=3.6,9.0Hz,1H),2.28-2.02(m,1H),1.93-1.56(m,1H).

[0846] Example 61. Synthesis of INSCoV-600D

[0847] Plan 30

[0848]

[0849] Step 1: at 70 ° C, under N2, a mixture of compound 1 (5g, 34.83mmol, 1eq, HCl) in ethyl formate (46.05g, 621.64mmol, 50mL, 17.85eq) and TEA (7.05g, 69.66mmol, 9.70mL, 2eq) was stirred for 12h. TLC (EA / PE=1 / 1) indicated that compound 1 (Rf=0.0) was consumed and a new spot (Rf=0.1) was formed. The mixture was filtered and washed with PE (10mL×3). The filtrate was concentrated in vacuo. The reaction was used for the next step without purification. Compound 2 (5g, crude product) as a white solid was obtained.

[0850] 1 H NMR (400 MHz, chloroform-d): δ = 8.11 (s, 1H), 6.92 (s, 1H), 4.40-4.24 (m, 1H), 3.01-2.91 (m, 2H), 2.63-2.46 (m, 2H).

[0851] Step 2: To a solution of compound 2 (2.0 g, 14.80 mmol, 1 eq) and TEA (1.50 g, 14.80 mmol, 2.06 mL, 1.0 eq) in DCM (20 mL) was added PPh (4.27 g, 16.28 mmol, 1.1 eq) and CCl (2.28 g, 14.80 mmol, 1.42 mL, 1.0 eq). The mixture was stirred at 45° C. under N for 12 h. TLC (PE / EA=3 / 1) indicated that compound 2 (Rf=0.1) was consumed and a small spot (Rf=0.6) was formed. The resulting mixture was evaporated in vacuo (<20° C.). At 20° C. under N, the residue was suspended in EtO (100 ml) for 12 h. The mixture was filtered, washed with EtO (50 mL×2), and the filtrate was concentrated in vacuo (<20° C.). The reaction was used in the next step without purification. Compound 3 (1.0 g, crude) was obtained as a yellow oil.

[0852] 1 H NMR (400MHz, DMSO-d6) δ = 4.39-4.20 (m, 1H), 3.22-3.06 (m, 2H), 3.00-2.83 (m, 2H).

[0853] Step 3: Synthesize INSCoV-600D according to the general procedure for the INSCoV series. Purification B: MTBE (20 mL) was added to the reaction mixture and cooled to 0 ° C for 12 h. The mixture was filtered and washed with MTBE (10 mL × 3) to obtain the product. The filter cake was concentrated in vacuo. INSCoV-600D (181.18 mg, 382.10 μmol, 41.30% yield, 97.402% purity) was obtained as a yellow solid, which was confirmed by HNMR, FNMR, LCMS, SFC and HPLC.

[0854] LCMS: Retention time: 0.869 min, (M+H) = 462.2. HPLC: Retention time: 1.657 min. SFC: Retention time: 1.255 min, 1.487 min. 1 H NMR (400MHz, DMSO-d6): δ = 8.99 (s, 1H), 8.78 (d, J = 6.0Hz, 1H), 8.51 (s, 2H), 8.46 (s, 1H), 7.72 (s, 1H), 7. 66(d,J=8.8Hz,2H),7.42(s,2H),6.05(s,1H),4.15-3.97(m,3H),3.01-2.82(m,2H),2.63-2.51(m,2H). 19 F NMR (377MHz, DMSO-d6): δ = -82.27 (d, J = 194.6Hz, 1F), -95.79 (d, J = 197.4Hz, 1F).

[0855] Example 62. Synthesis of INSCoV-600E

[0856] Plan 31

[0857]

[0858] Step 1: At 90 ° C, a mixture of 3-methyloxetane-3-amine (1g, 11.48mmol, 1eq) in ethyl formate (3g, 40.50mmol, 3.26mL, 3.53eq) was stirred for 16h. TLC (PE: EA = 1: 1) showed that the starting material (Rf = 0.4) was completely consumed, and a new spot (Rf = 0.2) was observed. The mixture was concentrated under reduced pressure. N- (3-methyloxetane-3-yl) formamide (1.3g, 11.29mmol, 98.37% yield) was obtained as a brown solid, which was detected by HNMR.

[0859] 1H NMR (400 MHz, CHLOROFORM-d): δ = 8.14 (d, J = 0.9 Hz, 1H), 7.27 (s, 1H), 4.81 (d, J = 6.4 Hz, 2H), 4.51 (d, J = 6.6 Hz, 2H), 1.69 (s, 3H).

[0860] Step 2: at -10 ℃, under N 2, to N- (3-methyloxetane-3-yl) formamide (1g, 8.69mmol, 1eq) and DIEA (5.61g, 43.43mmol, 7.56mL, 5eq) in DCM (100mL) in one-time addition POCl 3 (2.00g, 13.03mmol, 1.21mL, 1.5eq) is then heated to 25 ℃ and stirred for 2 hours. TLC (plate 1, PE: EA = 1: 1) shows that the starting material (Rf = 0.2) is completely consumed, and a new spot (Rf = 0.8) is observed. At 0 ℃, the mixture is poured into ice-saturated sodium bicarbonate solution (w / w = 1 / 1) (100mL) and stirred for 10min. The aqueous phase is extracted with DCM (100mL × 2) and dried over anhydrous Na 2 SO 4. At 20 ℃, the mixture is concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 40g The mixture was purified by silica flash column (0-50% ethyl acetate / petroleum ether gradient at 50 mL / min). The mixture was concentrated under reduced pressure at 20°C. TLC (plate 2, PE:EA=1:1, Rf=0.8). 3-isocyanato-3-methyl-oxetane (350 mg, 3.60 mmol, 41.49% yield) was obtained as a colorless oil.

[0861] Step 3: Synthesis of INSCoV-600E according to the general procedure for the INSCoV series. Purification A: The crude product was purified by preparative HPLC (column: Welch MLtimate XB-CN 250×50×10 μm; mobile phase: [hexane-IPA]; B%: 30%-70%, 15 min). 2-(N-(2-chloroacetyl)-4-oxazol-5-yl-anilino)-N-(3-methyloxetane-3-yl)-2-pyrimidin-5-yl-acetamide (105.94 mg, 228.59 μmol, 24.41% yield, 95.342% purity) was obtained as a yellow solid, which was detected by LCMS, HPLC and HNMR.

[0862] LCMS: retention time: 0.750 min, [M+H + ] = 442.2. HPLC: retention time: 1.580 min. 1H NMR (400MHz, DMSO-d6): δ = 8.99 (s, 1H), 8.84 (s, 1H), 8.52 (s, 2H), 8.47 (s, 1H), 7.73 (s, 1H), 7.66 (br d,J=8.6Hz,2H),7.54-7.33(m,2H),6.06(s,1H),4.56(dd,J=6.2,15.8Hz,2H),4.33-4.27(m,2H),4.18-4.00(m,2H),1.50(s,3H).

[0863] Example 63. N-(4-(1H-imidazol-5-yl)phenyl)-2-chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2- Synthesis of Oxo-1-(Pyrimidin-5-yl)ethyl)acetamide (INSCoV-600I)

[0864] INSCoV-600I was synthesized according to the general procedure for the INSCoV series. Purification A: The residue was dissolved in MeOH (2 mL) and purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 12%-32%, 6.5 min) and concentrated to remove MeCN, and the liquid was lyophilized to give a crude product. The residue was dissolved in DMF (2 mL) and purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 13%-33%, 6.5 min) and concentrated to remove MeCN, and the liquid was lyophilized to give a crude product. INSCoV-600I (13.3 mg, 25.58 μmol, 2.76% yield, 94.024% purity) was obtained as a yellow solid, which was confirmed by LCMS, HPLC, HNMR and FNMR.

[0865] LCMS: Retention time: 0.840 min, (M+H) = 489.2; Retention time: 0.724 min, (M+H) = 489.0. HPLC: Retention time: 1.767 min. 1 H NMR (400 MHz, methanol-d4): δ = 9.02 (d, J = 1.2 Hz, 1H), 8.94 (s, 1H), 8.57 (s, 2H), 7.96 (d, J = 1.2 Hz, 1H), 7.84-7.21 (m, 4H), 6.15 (s, 1H), 4.08-3.81 (m, 3H), 2.12-1.80 (m, 6H), 1.72-1.41 (m, 2H). 19 F NMR (376 MHz, methanol-d4): δ = -88.44 - -112.41 (m, 1F).

[0866] Example 64. Synthesis of INSCoV-600L

[0867] Plan 32

[0868]

[0869] Step 1: To a solution of 4-chloropyrimidine (500 mg, 3.31 mmol, 1 eq, HCl) and N-[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]acetamide (1.04 g, 3.97 mmol, 1.2 eq) in dioxane (5 mL) and H2O (1 mL) was added Pd(dppf)Cl2 (242.30 mg, 331.14 μmol, 0.1 eq) and NaHCO3 (834.57 mg, 9.93 mmol, 386.38 μL, 3.0 eq), which was purged with N2 three times and stirred at 100°C for 16 h. LCMS showed that 4-chloropyrimidine was consumed and the desired mass was detected. Water (20 ml) and EA (20 ml) were added, and the organic layer was washed with brine (20 ml), dried over Na2SO4 and concentrated in vacuo. The crude product was purified by column chromatography (SiO 2 , PE:EA 10:1 to 1:2). N-(4-pyrimidin-4-ylphenyl)acetamide (400 mg, 1.83 mmol, 55.29% yield, 97.596% purity) was obtained as a yellow solid, which was confirmed by HNMR and LCMS.

[0870] LCMS: retention time: 0.602 min, (M+H)=214.1. 1 H NMR (400 MHz, CHLOROFORM-d): δ = 9.16 (d, J = 1.1 Hz, 1H), 8.66 (d, J = 5.4 Hz, 1H), 8.06-7.95 (m, 2H), 7.67-7.56 (m, 3H), 7.44 (br s, 1H), 2.15 (s, 3H).

[0871] Step 2: At 25 ° C, HCl (10 mL) (2M) was added to a solution of N-(4-pyrimidin-4-ylphenyl)acetamide (350 mg, 1.64 mmol, 1 eq) in MeOH (10 mL), and it was stirred for 2 h at 70 ° C. LCMS showed that N-(4-pyrimidin-4-ylphenyl)acetamide was consumed, and the required mass was detected. It was poured into water (50 ml), and the pH of the mixture was adjusted to 9 by solid Na CO , EA (50 ml) was added, and the organic layer was washed with brine (50 ml), dried over Na SO and concentrated in vacuo. The crude product was used directly in the next step. 4-pyrimidin-4-ylaniline (280 mg, 1.64 mmol, 99.64% yield) was obtained as a yellow solid, which was confirmed by HNMR.

[0872] LCMS: retention time: 0.702 min, (M+H)=172.3. 1 H NMR (400MHz, DMSO-d6): δ=9.03 (d, J=1.3Hz, 1H), 8.62 (d, J=5.6Hz, 1H), 7.99-7.90 (m, 2H), 7.82 (dd, J=1.4, 5.5Hz, 1H), 6.72-6.61 (m, 2H), 5.81 (s, 2H).

[0873] Step 3: INSCoV-600L is synthesized according to the general procedure for the INSCoV series. Purification B: The crude product is ground with EtOH (1 ml) and filtered, and the filter cake is washed with PE (1 ml) and dried in vacuo. 2- (N- (2- chloroacetyl) -4- pyrimidin-4-yl-anilino) -N- (4,4- difluorocyclohexyl) -2- pyrimidin-5-yl-acetamide (144.91 mg, 279.56 μmol, 26.42% yield, 96.640% purity) as a yellow solid was obtained, which was confirmed by LCMS, HPLC, HNMR and FNMR.

[0874] LCMS: retention time: 0.850 min, (M+H) = 501.1. HPLC: retention time: 1.769 min. 1H NMR (400MHz, DMSO-d6) δ = 9.24 (s, 1H), 8.96 (s, 1H), 8.87 (d, J = 5.3Hz, 1H), 8.51 (s, 2H), 8.33 (br d, J = 7.3Hz, 1H), 8.24-8.02 (m, 3H), 7.53 (br s,2H),6.19-6.05(m,1H),4.16-4.00(m,2H),3.85(br s,1H),2.07-1.69(m,6H),1.62-1.46(m,1H),1.46-1.29(m,1H),1.17(t,J=7.2Hz,1H).

[0875] Example 65. Synthesis of INSCoV-600M

[0876] Plan 33

[0877]

[0878] Step 1: At 60 ° C, a mixture of 1-methylpiperidin-4-amine (2g, 17.51mmol, 1eq) in ethyl formate (12.97g, 175.15mmol, 14.09mL, 10eq) was stirred for 16h. LC-MS showed that the reaction was complete and the desired product was obtained. The mixture was concentrated under reduced pressure. N-(1-methyl-4-piperidinyl)formamide (2.45g, crude product) was obtained as a brown oil.

[0879] LCMS: retention time: 0.197 min, [M+H + ]=143.3.

[0880] Step 2: At 0 ° C, to a solution of N- (1- methyl -4- piperidinyl) formamide (1g, 7.03mmol, 1eq) and TEA (2.13g, 21.10mmol, 2.94mL, 3eq) in DCM (50mL) was added POCl (3.23g, 21.10mmol, 1.96mL, 3eq). The mixture was stirred for 2h at 20 ° C. LCMS showed that the reaction was complete and the desired product was obtained. At 0 ° C, the mixture was poured into saturated sodium bicarbonate (50mL). The aqueous phase was extracted with ethyl acetate (100mL × 3), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. 4-isocyanato-1-methyl-piperidine (120mg, crude product) was obtained as a brown oil, which was detected by LCMS.

[0881] LCMS: retention time: 0.697 min, [M+H + ]=125.2.

[0882] Step 3: INSCoV-600M was synthesized according to the general procedure for the preparation of the INSCoV series. The crude product was purified by preparative HPLC (column: Welch MLtimate XB-CN 250×50×10 μm; mobile phase: [hexane-IPA]; B%: 35%-75%, 15 min). 2-(N-(2-chloroacetyl)-4-oxazol-5-yl-anilino)-N-(1-methyl-4-piperidinyl)-2-pyrimidin-5-yl-acetamide (22.04 mg, 44.07 μmol, 4.71% yield, 93.756% purity) was obtained as a yellow solid, which was detected by LCMS, HPLC and HNMR.

[0883] LCMS: retention time: 0.824 min, [M+H + ] = 469.2. HPLC: retention time: 1.646 min. 1 H NMR (400 MHz, DMSO-d 6 ): δ=9.01-8.92(m,1H),8.57-8.43(m,3H),8.42-8.33(m,1H),7.71(s,1H), 7.68-7.59(m,2H),7.57-7.33(m,2H),6.07(s,1H),4.04(d,J=8.1Hz,2H),3 .73-3.63(m,1H),3.09-2.78(m,2H),2.69-2.64(m,1H),2.70-2.62(m,1H), 2.42-2.34(m,3H),1.89-1.67(m,2H),1.61-1.45(m,1H),1.42-1.28(m,1H).

[0884] Example 66. Synthesis of INSCoV-600O

[0885] Plan 34

[0886]

[0887] Step 1: at 25 ℃, to the mixture of 4-amino-3-methyl-benzoic acid (5.00g, 33.1mmol, 1.00eq) in DMF (50.0mL), NCS (4.42g, 33.1mmol, 1.00eq) was added once. The mixture was stirred for 1h at 100 ℃. LCMS showed that 4-amino-3-methyl-benzoic acid was consumed, and the required mass was detected. The mixture was poured into water (100mL) and then filtered. The filter cake was washed with water (50mL) and then vacuum-dried. It was not purified and used for the next step. 4-amino-3-chloro-5-methyl-benzoic acid (5.50g, 29.6mmol, 89.6% yield) was obtained as a brown solid, which was determined by HNMR.

[0888] LCMS: retention time: 0.702 min, [M+H + ]=186.1. 1 H NMR (400MHz, DMSO-d6): δ = 12.34 (s, 1H), 7.61 (d, J = 1.6Hz, 1H), 7.52 (s, 1H), 5.81 (s, 2H), 2.16 (s, 3H).

[0889] Step 2: To a solution of 4-amino-3-chloro-5-methyl-benzoic acid (3.00 g, 16.2 mmol, 1.00 eq) and methylamine hydrochloride (2.18 g, 32.3 mmol, 2.0 eq) in DMF (30.0 mL) at 20°C was added HATU (12.3 g, 32.3 mmol, 2.00 eq) and DIPEA (4.18 g, 32.3 mmol, 5.63 mL, 2.00 eq). The mixture was stirred at 20°C for 6 h. TLC (PE:EA = 1:1) showed that 4-amino-3-chloro-5-methyl-benzoic acid (Rf = 0.5) was consumed, and a new spot (Rf = 0.4) was observed. The mixture was poured into water (100 mL) and extracted with EA (50 mL x 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated in vacuo to yield a residue. The residue was purified by silica gel chromatography (PE:EA=1:1-0:1) to obtain 4-amino-3-chloro-N,5-dimethyl-benzamide (3.20 g, 16.1 mmol, 99.7% yield) as a white solid, which was identified by HNMR.

[0890] 1 H NMR (400 MHz, chloroform-d): δ = 7.58 (d, J = 1.8 Hz, 1H), 7.43-7.39 (m, 1H), 2.96 (s, 3H), 2.21 (s, 3H).

[0891] Step 3: At 20 ° C, under N2, a mixture of formic acid (405 mg, 8.81 mmol, 332 μ L, 3.50 eq) in acetic anhydride (308 mg, 3.02 mmol, 283 μ L, 1.20 eq) was added at once. The mixture was stirred for 10 min at 20 ° C. At 20 ° C, a solution of 4-amino-3-chloro-N, 5-dimethyl-benzamide (500 mg, 2.52 mmol, 1.00 eq) in DCM (5 mL) was added to the mixture. The resulting mixture was stirred for 3 h at 30 ° C. LCMS showed that 4-amino-3-chloro-N, 5-dimethyl-benzamide was consumed and the required mass was detected. The mixture was concentrated in vacuo to produce a residue. The residue was ground with EA (10 mL) and then filtered. The filter cake was vacuum dried. 3-Chloro-4-formamido-N,5-dimethyl-benzamide (263 mg, 1.16 mmol, 46.10% yield) was obtained as a yellow solid as confirmed by HNMR.

[0892] LCMS: retention time: 0.209 min, [M+H + ]=227.2. 1 H NMR (400MHz, DMSO-d6): δ=8.29 (d, J=1.3Hz, 1H), 7.76-7.68 (m, 1H), 2.77 (d, J=4.5Hz, 3H), 2.23 (s, 3H).

[0893] Step 4: To a solution of 3-chloro-4-formamido-N,5-dimethyl-benzamide (260 mg, 1.15 mmol, 1.00 eq) and TEA (116 mg, 1.15 mmol, 160 μL, 1.00 eq) in DCM (4.00 mL) was added POCl (176 mg, 1.15 mmol, 107 μL, 1.00 eq) at 0°C. The mixture was stirred at 20°C for 1 h. TLC (PE:EA=1:1) showed that 3-chloro-4-formamido-N,5-dimethyl-benzamide (Rf=0.1) was consumed, and a new spot (Rf=0.7) was observed. The mixture was diluted with DCM (50 mL) and then poured into NaHCO (50 mL). The resulting mixture was separated by a separatory funnel. The organic layer was dried over NaSO, filtered, and concentrated in vacuo to produce a residue. The residue was purified by silica gel chromatography (PE:EA=10:1-5:1) to obtain 3-chloro-4-isocyano-N,5-dimethyl-benzamide (60.0 mg, 288 μmol, 25.1% yield) as a yellow solid, which was determined by HNMR.

[0894] 1H NMR (400MHz, DMSO-d6): δ = 8.66 (d, J = 4.0Hz, 1H), 7.92 (d, J = 1.2Hz, 1H), 7.84 (s, 1H), 2.78 (d, J = 4.4Hz, 3H), 2.46 (s, 3H).

[0895] Step 5: Synthesis of INSCoV-600O according to the general procedure for the INSCoV series. Purification C: The residue was purified by silica gel chromatography (PE: EA = 10: 1-0: 1). 3-Chloro-4-[[2-(N-(2-chloroacetyl)-4-oxazol-5-yl-anilino)-2-pyrimidin-5-yl-acetyl] amino]-N, 5-dimethyl-benzamide (15.54 mg, 25.55 μmol, 13.33% yield, 91% purity) was obtained as a yellow solid, which was determined by HNMR, LCMS and HPLC.

[0896] LCMS: retention time: 0.823 min, [M+H + ] = 554.9. HPLC: retention time: 1.585 min. 1 H NMR (400MHz, DMSO-d6): δ = 10.20 (s, 1H), 9.00 (s, 1H), 8.62 (s, 2H), 8.57-8.51 (m, 1H), 8.46 (s, 1H), 7.8 1-7.63(m,5H),7.55-7.32(m,2H),6.33(s,1H),4.10(s,2H),2.78(d,J=4.4Hz,3H),2.33-2.23(m,3H).

[0897] Example 67. Synthesis of INSCoV-600R(2), 138.INSCoV-600R(2A) and 139.INSCoV-600R(2B) become

[0898] Plan 35

[0899]

[0900] Step 1: To a solution of compound 1 (0.5 g, 4.90 mmol, 1 eq) in THF (2 mL), MeOH (2 mL) and H2O (1 mL) was added LiOH * H2O (411.05 mg, 9.80 mmol, 2 eq). The reaction mixture was stirred for 1 h at 25 ° C. TLC (PE: EA = 3: 1) showed the formation of a new spot (Rf = 0.0). The reaction mixture was concentrated in vacuo to remove MeOH and THF. The mixture was then adjusted to pH = 2 by 1N HCl solution and extracted with EA (50 mL × 3). The combined organic phases were concentrated in vacuo (< 25 ° C). The reaction mixture was used for the next step without purification. Compound 2 (0.3 g, 3.41 mmol, 69.56% yield) was obtained as a colorless oil, which was confirmed by HNMR.

[0901] 1 H NMR (400MHz, DMSO-d6): δ=3.41-3.85(m,1H), 2.93-2.87(m,1H), 2.82-2.76(m,1H).

[0902] Step 2: INSCoV-600R (2) was synthesized according to the general procedure for the INSCoV series. The reaction was concentrated in vacuo. The crude product was ground with MTBE (20 mL) and washed with MTBE (10 mL × 2). The residue was diluted with MTBE (20 mL), washed with MTBE (10 mL × 2) and concentrated in vacuo. INSCoV-600R (2) (212.57 mg, 400.82 μmol, 43.33% yield, 91.161% purity) was obtained as a yellow solid, which was confirmed by HNMR, FNMR, LCMS, SFC and HPLC.

[0903] LCMS: retention time: 0.878 min, (M+H) = 484.3. SFC: retention time: 1.492 min, 2.151 min. 1 HNMR (400MHz, DMSO-d6) δ = 8.97 (d, J = 8.8Hz, 1H), 8.51 (d, J = 5.6Hz, 2H), 8.45 (d, J = 0.8Hz, 1H), 8.42-8.27 (m, 1H), 7.76-7.61 (m, 3H), 7.52-7.3 6(m,2H),6.24-5.96(m,1H),3.93-3.75(m,1H),3.17-3.08(m,1H),2.87 -2.75(m,1H),2.75-2.69(m,1H),2.05-1.69(m,6H),1.61-1.29(m,2H). 19F NMR(377MHz,DMSO-d6)δ=-87.22-103.36(m,1F)

[0904] Step 3: General procedure for the preparation of INSCoV-600R (2A) and INSCoV-600R (2B). The products were purified by chiral SFC (column: DAICEL CHIRALPAK AD (250 mm × 30 mm, 10 μm); mobile phase: [0.1% NH3·H2O INSCoV-600R (2) (0.1 g, 206.84 μmol, 1 eq) was isolated by 4% methanol, 4% ethanol, 4% ethanol, 0.5% ethanol, 0.6% ethanol, 0.7% ethanol, 0.8% ethanol, 0.9% ethanol, 1.0% ethanol, 0.9% ethanol, 1.2% ethanol, 0.8% ethanol, 0.9% ethanol, 1.0% ethanol, 0.8% ethanol, 1.2% ethanol, 0.8% ethanol, 1.0 ...

[0905] INSCoV-600R (2A): LCMS: retention time: 0.876 min, (M+H) = 484.3, HPLC: retention time: 1.601 min, SFC: retention time: 1.533 min, 1 H NMR (400MHz, DMSO-d6) δ = 8.96 (d, J = 1.2 Hz, 1H), 8.50 (s, 2H), 8.45 (d, J = 1.2 Hz,1H),8.36(d,J=7.6Hz,1H),7.71(s,1H),7.67(d,J=8.0Hz,2H),7.47(s, 2H),6.14(s,1H),3.90-3.80(m,1H),3.13-3.08(m,1H),2.87-2.81(m,1H), 2.77-2.71(m,1H),2.01-1.75(m,6H),1.62-1.47(m,1H),1.45-1.31(m,1H), 19 F NMR (377MHz, DMSO-d6) δ = 91.06 -95.14 (m, 1F), -96.46 - 100.47 (m, 1F).

[0906] INSCoV-600R (2A): LCMS: retention time: 0.874 min, (M+H) = 484.3, HPLC: retention time: 1.593 min, SFC: retention time: 2.139 min,1 H NMR (400MHz, DMSO-d6) δ=8.98(s,1H),8.51(s,2H),8.45(s,1H),8.31(d,J=7.6Hz,1H),7.71(s,1H),7.67(d,J=8.8Hz,2H),7.44(d,J=7.2Hz,2H ),6.08(s,1H),3.84-3.78(m,1H),3.13(s,1H),2.81-2.75(m,1H),2.75 -2.69(m,1H),2.01-1.73(m,6H),1.56-1.44(m,1H),1.40-1.30(m,1H), 19 F NMR (37MHz, DMSO-d6) δ = -93.34 (d, J = 234.6Hz, 1F), -96.19--99.53 (m, 1F).

[0907] Based on modeling and activity data, INSCoV-600R(2A) is expected to have structure.

[0908] Example 68. Synthesis of INSCoV-600X

[0909] Plan 36

[0910]

[0911] Step 1: To a solution of compound 1 (2 g, 9.80 mmol, 1 eq) and compound 2 (2.41 g, 11.76 mmol, 1.2 eq) in dioxane (20 mL) was added CsCO (9.58 g, 29.41 mmol, 3 eq) and Pd(dppf)Cl (1.43 g, 1.96 mmol, 0.2 eq). The reaction mixture was stirred at 80°C under N for 12 h. LCMS showed complete consumption of compound 1, and a peak with the desired mass was detected (Rt = 1.063 min). TLC (PE:EA = 1:1) showed complete consumption of compound 1 (Rf = 0.8), and the formation of three new spots (Rf = 0.02, Rf = 0.3, Rf = 0.5). The mixture was diluted with water (30 mL) and extracted with EtOAc (80 mL x 2). The organic layer was dried over anhydrous NaSO, filtered, and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 80g The residue was purified by silica flash column, 0-60% ethyl acetate / petroleum ether gradient at 100 mL / min and concentrated in vacuo to obtain compound 3 (3.5 g, crude) as a yellow solid, which was identified by HNMR and FNMR.

[0912] LCMS: retention time: 1.063 min, (M+H)=203.1, 1 H NMR (400MHz, DMSO-d6): δ = 8.81 (d, J = 2.4Hz, 1H), 8.47–8.41 (m, 1H), 7.99–7.83 (m, 1H),7.41–7.35(m,1H),7.34–7.28(m,1H),7.24(s,1H),5.10(s,2H),2.19(s,3H), 19 F NMR (377MHz, DMSO-d6): δ = -134.656 (m, 1F).

[0913] Step 2: At 25 ° C, under N2, a mixture of HCOOH (2.41g, 50.19mmol, 3.5eq) and Ac2O (1.76g, 17.21mmol, 1.61mL, 1.2eq) was stirred for 10min. At 0 ° C, a solution of compound 3 (2.9g, 14.34mmol, 1eq) in DCM (30mL) was added to the mixture. The resulting mixture was stirred for 3h at 25 ° C. LCMS showed that compound 3 was completely consumed, and a peak with the desired mass was detected (Rt = 0.797min). TLC (PE: EA = 1: 2) showed that compound 3 (Rf = 0.7) was left, and a spot (Rf = 0.5) was formed. The reaction mixture was diluted with water (20mL) and extracted with DCM (30mL × 3). The combined organic phase was dried over anhydrous Na2SO4 and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 80g The product was purified by silica flash column (0-100% ethyl acetate / petroleum ether gradient eluent at 100 mL / min). The combined organic phases were dried over anhydrous Na2SO4 and concentrated in vacuo. Compound 4 (1.5 g, 6.52 mmol, 45.43% yield) was obtained as a white solid, which was confirmed by HNMR.

[0914] LCMS: retention time: 0.797 min, (M+H)=231.2, 1 H NMR (400MHz, DMSO-d6): δ = 9.77 (s, 1H), 8.93 (d, J = 1.8Hz, 1H), 8.62-8.56 (m 1H), 8.31 (s, 1H), 8.14-8.08 (m, 1H), 7.56-7.46 (m, 3H), 2.28 (s, 3H).

[0915] Step 3: To a solution of compound 4 (1.2 g, 5.21 mmol, 1 eq) in DCM (12 mL) was added TEA (527.40 mg, 5.21 mmol, 725.45 μL, 1 eq), PPh 3 (1.50 g, 5.73 mmol, 1.1 eq) and CCl 4 (801.73 mg, 5.21 mmol, 501.08 μL, 1 eq). The reaction mixture was stirred for 12 h at 45 ° C under N 2. LCMS showed that compound 4 was residual (Rt = 0.807 min) and the required mass was not detected. TLC (PE: EA = 1: 1) showed the formation of three new spots (Rf = 0.2, Rf = 0.3, Rf = 0.7). The mixture was diluted with water (10 mL) and extracted with DCM (20 mL × 3). The organic layer was dried over anhydrous Na 2 SO 4, filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography ( 80g The residue was purified by silica flash column, 0-80% ethyl acetate / petroleum ether gradient at 100 mL / min and concentrated in vacuo to obtain compound 5 (1 g, 4.71 mmol, 90.41% yield) as a white solid, which was confirmed by HNMR and FNMR.

[0916] LCMS: retention time: 0.805 min, (M+H)=231.1, 1 H NMR (400MHz, DMSO-d6): δ = 8.97 (d, J = 2.4Hz, 1H), 8.66-8.60 (m, 1H), 8.19-8.1 3(m,1H),7.80(d,J=10.8Hz,1H),7.73(s,1H),7.55-7.49(m,1H),2.49(s,3H), 19 F NMR (377MHz, DMSO-d6): δ = -119.049 (m, 1F).

[0917] Step 4: INSCoV-600X was synthesized according to the general procedure for the preparation of the INSCoV series. Purification B: MTBE (20 mL) was added to the reaction mixture, filtered and washed with MTBE (10 mL × 3) to obtain a crude product. The residue was ground with MTBE (20 mL), filtered and washed with MTBE (10 mL × 3). The filter cake was concentrated in vacuo. INSCoV-600X (301.95 mg, 517.73 μmol, 55.97% yield, 95.5% purity) was obtained as a yellow solid, which was confirmed by HNMR, FNMR, LCMS, SFC and HPLC.

[0918] LCMS: retention time: 0.774 min, (M+H)=557.1, HPLC: retention time: 1.522 min, SFC: retention time: 1.106 min, 3.019 min, 1 H NMR (400MHz, DMSO-d6): δ = 10.04 (s, 1H), 9.02 (s, 1H), 8.94 (d, J = 1.8Hz, 1H), 8.62 (s, 2H), 8.62–8.56 (m, 1H), 8.46 (s, 1H),8.15–8.09(m,1H),7.73(s,1H),7.67(d,J=8.4Hz,2H),7.61-7.38(m,5H),6.38(s,1H),4.10(s,2H),2.28(s,3H), 19 F NMR (377MHz, DMSO-d6): δ = -119.55 (s, 1F).

[0919] Example 69. N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyrimidin-5-yl)ethyl)-N-(4- Synthesis of (Isoxazol-5-yl)phenyl)oxirane-2-carboxamide (INSCoV-600Y)

[0920] INSCoV-600Y was synthesized according to the general procedure for the INSCoV series. Purification A: The residue was dissolved in DMF (2 mL) and purified by preparative HPLC (column: Waters Xbridge 150×25 mm×5 μm; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 20%-50%, 9 min) and diluted with water (20 mL). The liquid was lyophilized to produce the product. The residue was dissolved in DMF (2 mL) and purified by preparative HPLC (column: 3-Phenomenex Luna C18 75×30 mm×3 μm; mobile phase: [water (0.05% HCl)-ACN]; B%: 28%-48%, 6.5 min) and diluted with water (20 mL). The liquid was lyophilized to produce the crude product (unstable in acid), which was confirmed by LCMS. INSCoV-600Y (31.65 mg, 59.87 μmol, 6.47% yield, 91.460% purity) was obtained as a white solid, which was confirmed by LCMS, HPLC, HNMR and FNMR.

[0921] LCMS: retention time: 0.893 min, (M+H)=484.3, HPLC: retention time: 1.724 min, 1H NMR (400MHz, DMSO-d6): δ=9.04-8.90(m,1H),8.69-8.63(m,1H),8.58-8.47(m,2H),8.42-8.27(m,1H),7.86-7.80(m,2H),7.65-7.34(m,2 H),7.08-6.99(m,1H),6.19-6.07(m,1H),3.93-3.69(m,1H),3.15-3.09(m,1H),2.89-2.70(m,1H),2.03-1.71(m,6H),1.62-1.27(m,2H), 19 FNMR (377MHz, DMSO-d6): δ = -87.29-105.18 (m, 1F).

[0922] Example 70. 2-Chloro-N-(2-((4,4-difluorocyclohexyl)amino)-2-oxo-1-(pyridazin-4-yl)ethyl)- Synthesis of N-(4-(thiazol-5-yl)phenyl)acetamide (INSCoV-601F)

[0923] Compound 1 (80.43 mg, 851.12 μmol, 95.75 μL, 1eq) and compound 3 (92.00 mg, 851.12 μmol, 1eq) were added to a solution of compound 4 (150 mg, 851.12 μmol, 1eq) and compound 2 (123.54 mg, 851.12 μmol, 1eq) in CF3CH2OH (6 mL). The reaction mixture was stirred for 1 h at 25 ° C. LCMS showed that reactant 1 was consumed, and a peak with the desired mass was detected. The reaction was concentrated in vacuo. The crude product was ground and filtered with PE / EA=20 / 1 (20 mL*3). INSCoV-601F (224.12 mg, 417.59 μmol, 49.06% yield, 94.273% purity) was obtained as a brown solid, which was confirmed by HNMR, FNMR, LCMS, HPLC and SFC.

[0924] LCMS: retention time: 0.880 min, (M+H)=506.3, HPLC: retention time: 1.899 min, SFC: retention time: 0.499 min, 1.259 min, 1H NMR (400MHz, DMSO-d6): δ = 9.10 (d, J = 0.6Hz, 1H), 9.08-9.04 (m, 1H), 9.15-8 .98(m,1H),8.37(d,J=7.8Hz,1H),8.33(d,J=0.6Hz,1H),7.63(d,J=8.8Hz, 2H),7.46-7.37(m,3H),6.06(s,1H),4.16-4.01(m,2H),3.89-3.77(m,1H), 1.97-1.86(m,4H),1.80-1.76(m,2H),1.55-1.48(m,1H),1.41-1.40(m,1H), 19 F NMR (377MHz, DMSO-d6): δ = -87.22--104.30 (m, 1F).

[0925] Example 71. Synthesis of INSCoV-601J and INSCoV-601J(2)

[0926] Plan 37

[0927]

[0928] Step 1: Synthesize INSCoV-601J according to the general procedure for the INSCoV series. Purification B: The mixture was concentrated in vacuo. The crude product was dissolved in METB (10 mL), stirred for a while, and the filter cake was concentrated in vacuo. INSCoV-601J (250 mg, 427.35 μmol, 46.20% yield, 86.489% purity) was obtained as a yellow solid, which was confirmed by HNMR, LCMS, SFC, and HPLC.

[0929] LCMS: retention time: 0.844 min, (M+H)=506.1, HPLC: retention time: 2.000 min, SFC: retention time: peak: 0.491 min, peak 2: 1.545 min, 1 H NMR (400MHz, DMSO-d6): δ=9.19-9.02(m,2H),8.60(d,J=1.8Hz,1H),8.39(d,J=7.6Hz,1H),7.80(d,J=1.8Hz,1H),7.70(d, J=8.8Hz,2H),7.57-7.37(m,3H),6.07(s,1H),4.17-4.09(m,1H),3.85-3.74(m,1H),2.07-1.70(m,8H),1.57-1.40(m,2H).

[0930] Step 2: INSCoV-601J (100 mg, 197.64 μmol, 1 eq) was chiralized by SFC. The solution was concentrated in vacuo. INSCoV-601J (2) (54.54 mg, 104.52 μmol, 52.88% yield, 96.965% purity) was obtained as a yellow solid. It was confirmed by HNMR, FNMR, LCMS, and HPLC; SFC showed ee% = 83.00%.

[0931] LCMS: retention time: 0.832 min, (M+H) = 506.1, SFC: retention time: 1.564 min, ee% = 83.00%, HPLC: retention time: 1.994 min, 1 H NMR (400MHz, DMSO-d6): δ=9.14-9.04(m,2H),8.59(d,J=1.8Hz,1H),8.38(br d,J=7.5Hz,1H),7.80(d,J=1.8Hz,1H),7.70(br d,J=8.8Hz,2H),7.49(br d,J=7.3Hz,2H),7.39(dd,J=2.5,5.3Hz,1H),6.07(s,1H),4.21-4.02(m,2H),3.82(br dd,J=2.5,3.4Hz,1H),2.09-1.68(m,8H),1.55-1.37(m,2H). 19 F NMR (376MHz, DMSO-d6): δ=-92.54--94.15(m,1F),-96.48--98.66(m,1F).

[0932] Example 72. Synthesis of INSCoV-612

[0933] Plan 38

[0934]

[0935] INSCoV-612 was synthesized according to the general procedure for the INSCoV series. Purification B: H2O (75 mL) was added to the mixture and extracted with EA (30 mL×3). The organic phase was washed with brine (50 mL), dried over anhydrous Na2SO4, and concentrated in vacuo to produce a yellow residue. The crude product was triturated with EtOH (10 mL) for 30 min at 25°C to produce INSCoV-612 (170 mg, 342.38 μmol, 37.01% yield, 100% purity) obtained as a yellow solid.

[0936] LCMS: retention time: 0.752 min, (M+H)=497.1, HPLC: retention time: 2.495 min, 1 H NMR (400MHz, DMSO-d6): δ = 9.10 (s, 1H), 8.99 (s, 1H), 8.55-8.50 (m, 2H), 8.37-8.31 (m, 2H), 7.65 (br d, J = 8.6Hz, 2H), 7.38 (br s,2H),6.14-6.07(m,1H),3.90-3.82(m,1H),3.65(s,2H),2.07-2.01(m,1H),1.97-1.81 (m,4H),1.80-1.71(m,1H),1.60-1.50(m,1H),1.45-1.33(m,1H),1.18(t,J=7.1Hz,1H), 19 F NMR: (377MHz, DMSO-d6): δ=-92.63--93.92(m,1F),-97.01--99.16(m,1F).

[0937] Pharmaceutical composition

[0938] Example A-1: ​​Parenteral Pharmaceutical Composition

[0939] To prepare a parenteral pharmaceutical composition suitable for administration by injection (e.g., subcutaneous, intravenous), 1-1000 mg of a water-soluble salt of a compound described herein, or a pharmaceutically acceptable salt or solvate thereof, is dissolved in sterile water and then mixed with 10 mL of 0.9% sterile saline. A suitable buffer and, optionally, an acid or base are added to adjust the pH. The mixture is incorporated into a dosage unit form suitable for administration by injection (i.e., subcutaneous SC injection).

[0940] Example A-2: Oral Solution

[0941] To prepare a pharmaceutical composition for oral delivery, a sufficient amount of a compound described herein, or a pharmaceutically acceptable salt thereof, is added to water (with optional solubilizer(s), optional buffer(s), and taste-masking excipients) to provide a 20 mg / mL solution.

[0942] Example A-3: Oral Tablet

[0943] Tablets are prepared by mixing 20-50% by weight of a compound described herein or a pharmaceutically acceptable salt thereof, 20-50% by weight of microcrystalline cellulose, 1-10% by weight of low-substituted hydroxypropyl cellulose, and 1-10% by weight of magnesium stearate or other suitable excipients. Tablets are prepared by direct compression. The total weight of the compressed tablet is maintained at 100-500 mg.

[0944] Example A-4: Oral capsule

[0945] To prepare a pharmaceutical composition for oral delivery, 1-1000 mg of a compound described herein or a pharmaceutically acceptable salt thereof is mixed with starch or other suitable powder blends. The mixture is incorporated into oral dosage units suitable for oral administration, such as hard gelatin capsules.

[0946] In another embodiment, 1-1000 mg of a compound described herein, or a pharmaceutically acceptable salt thereof, is placed into a size 4 capsule or a size 1 capsule (hydroxypropyl methylcellulose or hard gelatin), and the capsule is sealed.

[0947] Biological Examples

[0948] Example B-1: In vitro assay (SARS-CoV-2 M pro Enzymatic assay)

[0949] Clone C-His6-tagged SARS-CoV-2M PRO (NC_045512), expressed in E. coli and purified by WuXi. The substrate Dabcyl-KTSAVLQ||SGFRKME-(Edans) was synthesized by Genscript. The assay buffer contained 20 mM Tris-HCl (pH = 7.3), 100 mM NaCl, 1 mM EDTA, 5 mM TCEP, and 0.1% BSA. PRO In the enzymatic assay, M pro The final concentrations of protein and substrate were 25nM and 25μM, respectively. Reference compound GC376 was provided by WuXi AppTec and was included in each plate to ensure assay robustness. Test compounds were tested in duplicate with a single dose or 10 dose titrations. Compounds were added to duplicate wells of an assay plate (384-well format) using ECHO. For single-dose experiments, the final concentration was 10μM. For full dose response experiments, samples were serially diluted 3-fold from 25uM to obtain 10 doses and added to duplicate wells of the assay plate. The final concentrations (μM) of each compound were 25, 8.33, 2.778, 0.926, 0.309, 0.103, 0.034, 0.011, 0.0038, and 0.0013. M was titrated using Multidrop. PRO Protein (25 μL, 30 nM) was added to the assay plate containing the test compound. PROProtein was pre-incubated for 30 min. Then, substrate (5 μL, 150 μM) was added to the assay plate. For the 100% inhibition control (HPE, high percentage effect), 1 μM GC376 was added. For the non-inhibition control (ZPE, zero percentage effect), the same volume of DMSO was added. The final DMSO concentration was 1%. Each activity test point had a relevant background control without enzyme to remove the fluorescence interference of the compound. After incubation at 30 ° C for 60 min, the plate was analyzed using a microplate reader M2e (SpectraMax) at E x / E m =340nm / 490nm to detect the fluorescence signal (RFU).

[0950] Inhibitory activity was calculated using the following formula, and IC was calculated using the % inhibition data. 50 value.

[0951] Inhibition % = ((CPD-BG HPE )-(ZPE-BG ZPE )) / ((HPE-BG HPE )-(ZPE-BG ZPE ))×100

[0952] Among them, HPE is the high percentage effect control (1 μM GC376+enzyme+substrate); ZPE is the zero percentage effect control (enzyme+substrate, no compound); CPD is the compound activity test well (compound+enzyme+substrate); and BG is the background control well (no enzyme).

[0953] The IC of the compounds was calculated using GraphPad Prism software using a nonlinear regression model of log(inhibitor) versus response with variable slope (four parameters). 50 value.

[0954] Representative biochemical data are presented in Table 2 , and representative biochemical curves are shown in Figure 2 middle.

[0955] Table 2. In vitro potency data

[0956]

[0957]

[0958]

[0959]

[0960] Example B-2: Cell-based in vitro antiviral assay (in vivo SARS-CoV-2 IFA)

[0961] SARS-CoV-2 was provided by the Korea Centers for Disease Control and Prevention (KCDC). Vero cells were obtained from ATCC and maintained in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% FBS and 1% antibiotic-antimycotic solution. DMEM supplemented with 2% FBS and 1% antibiotic-antimycotic solution was used as the assay medium. The main reagents used in this assay were anti-SARS-CoV-2N protein antibody, Alexa Fluor 488 goat anti-rabbit IgG (H+L) secondary antibody, and Hoechst 33342. A ten-point dose-response curve (DRC) was generated for each compound. 24 h before the experiment, Vero cells were plated at 1.2 × 10 4 Cells / well were seeded in black 384-well μClear plates (Greiner Bio-One). For viral infection, SARS-CoV-2 was added at a multiplicity of infection (MOI) of ~0.0125. At 24 hpi, the cells were fixed with 4% paraformaldehyde and analyzed by immunofluorescence. The images obtained were analyzed using software to quantify the number of cells and infection rate, and the antiviral activity was normalized to the positive (mock) and negative (0.5% DMSO) controls in each assay plate. Using XLfit 4 software or Prism, the DRC was fitted by a sigmoidal dose-response model using the following equation:

[0962] Y=Bottom+(Top-Bottom) / (1+(IC 50 / X)Hill slope

[0963] Calculating EC 50 and CC 50 The values ​​are shown in Table 3.

[0964] Table 3. Results of cell-based in vitro assays for live SARS-CoV-2.

[0965]

[0966]

[0967] EC was calculated in the case of remdesivir as a positive control 50 and CC 50 The structure of remdesivir is listed below.

[0968]

[0969] Example B-3: ADME Characterization

[0970] Microsomal stability assessment

[0971] The microsomal stability of the compounds from Table 4 was assessed as follows: Working solutions of the test and control compounds (testosterone, diclofenac, and propafenone) were prepared. The appropriate amount of NADPH powder (β-nicotinamide adenine dinucleotide phosphate, reduced form, tetrasodium salt, NADPH·4Na, catalog number 00616; Chem-Impex International) was weighed and diluted into a 10 mM MgCl2 solution (working solution concentration, 10 units / ml; final concentration in the reaction, 1 unit / ml). Microsomal working solutions of appropriate concentrations were prepared using 100 mM PB (human: HLM, catalog number 452117, Corning; CD-1 mouse: MLM, catalog number M1000, Xenotech). Cold ACN containing 100 ng / ml tolbutamide and 100 ng / ml labetalol as internal standards (IS) was used as the stop solution. Compound or control working solution (10 μ l / well) is added in all plates (T0, T5, T10, T20, T30, T60 and NCF60), except matrix blank. By Apricot, the microsomal solution (80 μ l / well) through distribution is added in each plate, and at 37 ℃, the mixture of microsomal solution and compound is hatched for about 10min. After preheating, by Apricot, the NADPH regeneration system (10 μ l / well) through distribution is added in each plate to start reaction. Then, at 37 ℃, hatch solution. Then add stop solution (300 μ l / well, 4 ℃) to terminate reaction. The sampling plate was vibrated for about 10 minutes. At 4 ℃, at 4,000rpm, the sample was centrifuged for 20min. While centrifuged, a new 8×96-well plate was loaded with 300 μl of HPLC water, followed by transfer of 100 μl of supernatant and mixing for liquid chromatography-tandem mass spectrometry (LC / MS / MS).

[0972] CYP inhibition assay

[0973] The following reagents / reactants were used in the assay: water purified by ELGA Lab purification system, buffer solutions: PB (100 mM), MgCl2 (33 mM), organic solvents were AR or HPLC grade, CYP substrates: phenacetin (10 μM, 25 μL, 1A2), diclofenac (5 μM, 25 μL, 2C9), S-mephenytoin (30 μM, 75 μL, 2C19), dextromethorphan (5 μM, 12,5, 2D6) and midazolam (2 μM, 10 μL, 3A4), stock solutions were prepared in MeOH (20, 10, 20, 20 and 10 mM, respectively). Positive controls: α-naphthoflavone, sulfaphenazole, (+)-N-3-benzylnirvanol, quinidine, and ketoconazole, final concentrations of 3 μM in MeOH (90 μL), stock solutions were prepared in DMSO (3 mM for all inhibitors). Test compounds were evaluated using five concentration points ranging from 15 to 5,000 μM. Human liver microsomes (Cat. No. 452117, Corning) were used at a concentration of 0.253 mg / mL (PB: 44,431 mL; volume of microsomes: 569 mL), while NADPH (Cat. No. 00616, Chem-impex international) was used at a concentration of 10 mM (MgCl2: 20,0 mL, 33 mM). Main procedure: Prepare working solutions (100×) of test compounds and standard inhibitors, remove microsomes from a –80°C freezer, thaw on ice, date, and return to the freezer immediately after use. Add substrate (20 μL) and PB (20 μL) to the corresponding wells and a blank well, respectively. Add test compounds (2 μL) and positive control working solutions to the corresponding wells, followed by solvent (2 μL) to the inhibitor-free wells and the blank wells. Add HLM working solution (158 μL) to all wells of the incubation plate. Preheat the plate in a water bath at 37°C for approximately 10 minutes, followed by the addition of NADPH (20 μL) to all incubation wells. Mix the CYPs and incubate at 37°C in a water bath for 10 minutes. At the time point, stop the reaction by adding 400 μL of cold stop solution (200 ng / mL tolbutamide and labetalol in ACN). The samples were centrifuged at 4000 rpm for 20 minutes to precipitate the proteins. The supernatant (200 μL) was transferred to 100 μL HPLC water and shaken for 10 min before the samples were subjected to LC / MS / MS analysis.XL fit was used to plot the percentage of vehicle control versus test compound concentration and for nonlinear regression analysis of the data. IC was determined using a 3 or 4 parameter logistic equation. 50 When the inhibition percentage at the highest concentration (50 μM) is less than 50%, the IC 50 Values ​​are reported as ">50 μM".

[0974] Representative CYP P450 data are described in Table 4.

[0975] Table 4. Metabolic (microsomal) stability (MLM and HLM) and small CYP P450 panel assessment.

[0976]

[0977]

[0978] Example B-4: Permeability study

[0979] Caco-2 permeability

[0980] Caco-2 cells purchased from ATCC were plated at 1 × 105 cells / cm 2 Inoculate onto a polyethylene film (PET) in a 96-well Corning plug-in plate. Renew the culture medium every 4-5 days until the 21st to 28th day to form a confluent cell monolayer. The transport buffer is HBSS with 10mM HEPES at pH=7.40±0.05. Test compounds (2.00 μM) and digoxin (10.0 μM) were tested in duplicate in both directions, while nadolol and metoprolol were also tested in duplicate in the direction A to B at 2.00 μM. The final DMSO concentration was adjusted to less than 1%. In a CO2 incubator, at 37±1°C, at 5% CO2, at saturated humidity, without shaking, the plate was incubated for 2h. All samples were then mixed with acetonitrile containing internal standards and centrifuged at 3200×g for 10min. For nadolol and metoprolol, 100 μL of supernatant solution was diluted with 300 μL of distilled water for LC-MS / MS analysis. For digoxin and test compounds, 100 μL of supernatant solution was diluted with 100 μL of distilled water for LC-MS / MS analysis. The concentrations of the test and control compounds in the starting solution, donor solution, and receiver solution were quantified using the peak area ratio of the analyte / internal standard by the LC-MS / MS method. Following the transport assay, the Caco-2 cell monolayer integrity was determined using a lucifer yellow exclusion assay. The apparent permeability coefficient Papp (cm / s) was calculated using the following equation:

[0981] P app =(dCr / dt)×V r / (A×C0)

[0982] where dCr / dt is the cumulative concentration of the compound in the receptor compartment over time (μM / s); V r is the volume of solution in the receiver compartment (0.075 mL on the apical side and 0.25 mL on the basolateral side); A is the transport surface area, i.e., the area of ​​the monolayer is 0.0804 cm 2 ; C0 is the initial concentration in the donor compartment (μM).

[0983] Calculate the effluent ratio using the following equation:

[0984] Outflow ratio = P app (BA) / P app (AB)

[0985] Where V d is the volume in the donor chamber (0.075 mL on the apical side and 0.25 mL on the basolateral side); C d and C r are the final concentrations of transported compounds in the donor and acceptor compartments, respectively.

[0986] Data analysis was performed in a manner similar to the procedure described for the CYP inhibition assay and is shown in Table 5.

[0987] PAMPA permeability assay

[0988] 2.6g KH2PO4 and 18.5g K3PHO4×3H2O are dissolved in 1000mL of ultrapure water and mixed thoroughly. 1M sodium hydroxide or 1M hydrochloric acid is used to adjust the pH to 7.40±0.05. A 0.2mM working solution is prepared by diluting a 10mM stock solution with DMSO. A 10μM donor solution (5% DMSO) is prepared by diluting 20μL of working solution with 380μL PBS. 150μL of 10μM donor solution is added to each well of the donor plate, the PVDF membrane of which is pre-coated with 5μL of a 1% lecithin / dodecane mixture. Prepared in duplicate. 300μL of PBS is added to each well of the PTFE acceptor plate. The donor plate and the acceptor plate are combined and incubated for 4h at room temperature under shaking at 300rpm. Preparation of T0 samples: 20 μL of donor solution was transferred to a new well, followed by addition of 250 μL PBS (DF: 13.5), 130 μL ACN (containing internal standards) as T0 samples. Preparation of acceptor samples: The plate was removed from the incubator. 270 μL of solution was transferred from each acceptor well and mixed with 130 μL ACN (containing internal standards) as acceptor samples. Preparation of donor samples: 20 μL of solution was transferred from each donor well and mixed with 250 μL PBS (DF: 13.5), 130 μL ACN (containing internal standards) as donor samples. Acceptor and donor samples were all analyzed by LC-MS / MS. The equation for determining the permeation rate (Pe) is shown below:

[0989]

[0990] [drug] 平衡 =([drug] 供体 ×V D +[Drugs] 受体 ×V A ) / (V D +V A ),

[0991] V D =0.15mL; V A =0.30mL; area =0.28cm 2 ; time = 14400s,

[0992] [drug] 受体 =(A a / A i ×DF) 受体 ;[drug] 供体 =(A a / A i ×DF) 供体 ,

[0993] Among them: A a / A: peak area ratio of analyte to internal standard; DF: dilution factor.

[0994] Representative p...

Claims

1. A compound having a structure of formula (X) or a pharmaceutically acceptable salt thereof: in, B1 is the key; B is a bond or a C1-C4 alkylene linker, wherein the alkylene is optionally substituted with C1-C6 alkyl or halogen; R1 is a haloacetyl group; R3 is pyrimidinyl; R4 is phenyl, 5- to 6-membered heteroaryl, C3-C8 cycloalkyl or 4-7-membered heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace; R5 is H or C1-C6 alkyl; R 11 is C1-C6 alkyl, C1-C6 alkoxy, phenyl or 5- to 6-membered heteroaryl, wherein each of said alkyl, alkoxy, phenyl or heteroaryl is optionally substituted with C1-C6 alkyl or halogen; R 15a , R 15b , R 15c and R 15d Each is independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; R 16 is H or C1-C6 alkyl; and Each R 19 are independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, -S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C3-C8 cycloalkyl sulfone, C1-C6 alkyl sulfone and phenyl sulfone; The “4-7 membered heterocycloalkyl” is a 4-7 membered heterocycloalkyl having one or two heteroatoms selected from nitrogen, oxygen and sulfur, the “5- to 6-membered heteroaryl” is a 5- to 6-membered heteroaryl having one or two heteroatoms selected from nitrogen, oxygen and sulfur, and the sulfur atom in the “4-7 membered heterocycloalkyl” is optionally oxidized.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of formula (XA):

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure of formula (XB):

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of formula (XI): in, R1 is a haloacetyl group; R3 is pyrimidinyl; R4 is phenyl, 5- to 6-membered heteroaryl, C3-C8 cycloalkyl or 4-7-membered heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace; R5 is H or C1-C6 alkyl; R 11 is C1-C6 alkyl, C1-C6 alkoxy, phenyl or 5- to 6-membered heteroaryl, wherein each of said alkyl, phenyl or heteroaryl is optionally substituted with one, two or three C1-C6 alkyl or halogen; R 15a and R 15c Each is independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; Each R 19 are independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 alkyl),- N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, - C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, - S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkane phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C3-C8 cycloalkyl sulfone, C1-C6 alkyl sulfone and phenyl Sulfone.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of formula (XI): in, R1 is a haloacetyl group; R3 is pyrimidinyl; R4 is a substituted C3-C8 cycloalkyl or an optionally substituted 4-7 membered heterocycloalkyl, wherein when substituted, each of which is replaced by one, two, three or four R 19 replace; R5 is H or C1-C6 alkyl; R 11 is C1-C6 alkyl, C1-C6 alkoxy, phenyl or 5- to 6-membered heteroaryl, wherein each of said alkyl, phenyl or heteroaryl is optionally substituted with one, two or three C1-C6 alkyl or halogen; R 15a and R 15c Each is independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; Each R 19 are independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, -S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C1-C6 alkyl sulfone and phenyl sulfone.

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of formula (XI): in, R1 is a haloacetyl group; R3 is pyrimidinyl; R4 is phenyl, 5- to 6-membered heteroaryl, C3-C8 cycloalkyl or 4-7-membered heterocycloalkyl, each of which is optionally substituted by one, two, three or four R 19 replace; R5 is H or C1-C6 alkyl; R 11 is phenyl or 5- to 6-membered heteroaryl, wherein each of said phenyl or heteroaryl is optionally substituted with one, two or three C1-C6 alkyl or halogen; R 15a and R 15c Each is independently H, amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; Each R 19 are independently selected from oxo, halogen, -CN, -NH2, -NH(C 1-6 alkyl),- N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, - C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, - S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkane phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C1-C6 alkyl sulfone and phenyl sulfone.

7. The compound or pharmaceutically acceptable salt thereof according to any one of claims 4 to 6, wherein the compound has the structure of formula (XIA):

8. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein B is a C1-C4 alkylene linker.

9. The compound of claim 8 or a pharmaceutically acceptable salt thereof, wherein B is a C2 or C3 alkylene linker.

10. The compound of any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, wherein B is a bond.

11. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein the compound has a structure of formula (XII), or a pharmaceutically acceptable salt thereof: in, Y2 and Y4 are each N; and Y1 and Y3 are CH.

12. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R5 is C1-C6 alkyl.

13. The compound or pharmaceutically acceptable salt thereof according to claim 12, wherein the alkyl group is methyl or ethyl.

14. The compound of any one of claims 1-6, or a pharmaceutically acceptable salt thereof, wherein R5 is H.

15. The compound of claim 11 or a pharmaceutically acceptable salt thereof, wherein the compound has a structure of formula (XIIA), 16. The compound according to claim 11 or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of formula (XIIB):

17. The compound of claim 2 or 3, or a pharmaceutically acceptable salt thereof, wherein the compound has a stereochemical purity of at least 80%.

18. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 15a is H; and R 15c It is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy.

19. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 15a is amino, halogen, -CN, -OH, -OCF3, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy; and R 15c It's H.

20. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, wherein R 15a and R 15c Each is H.

21. A compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 5, wherein R 11 is C1-C6 alkyl or C1-C6 alkoxy, optionally substituted by one, two or three C1-C6 alkyl or halogen.

22. The compound of claim 21 or a pharmaceutically acceptable salt thereof, wherein the alkyl group is methyl, ethyl or tert-butyl.

23. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 6, wherein R 11 is a 5- to 6-membered heteroaryl group, optionally substituted by one, two or three C1-C6 alkyl groups or halogen.

24. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein the heteroaryl group is a 5-membered heteroaryl group.

25. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein the heteroaryl group is furan, thiophene, oxazole, thiazole, isoxazole, triazole, oxadiazole or thiadiazole.

26. The compound of claim 23 or a pharmaceutically acceptable salt thereof, wherein R 11 is an unsubstituted 5- to 6-membered heteroaryl group.

27. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 6, wherein R4 is optionally replaced by one, two or three R 19 Substituted 4-7 membered heterocycloalkyl.

28. The compound of claim 27 or a pharmaceutically acceptable salt thereof, wherein each R 19 are independently halogen, oxo, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, -S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C1-C6 alkyl sulfone and phenyl sulfone.

29. The compound of claim 28 or a pharmaceutically acceptable salt thereof, wherein each R 19 are independently halogen.

30. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 6, wherein R4 is a C3-C8 cycloalkyl group, optionally substituted by one, two or three R 19 replace.

31. The compound of claim 30 or a pharmaceutically acceptable salt thereof, wherein each R 19 are independently halogen, oxo, -CN, -NH2, -NH(C 1-6 Alkyl), -N(C 1-6 Alkyl)2, -OH, -CO2H, -CO2-C 1-6 Alkyl, -C(=O)NH2, -C(=O)NH(C 1-6 alkyl), -C(=O)N(C 1-6 Alkyl)2, -S(=O)2NH2, -S(=O)2NH(C 1-6 alkyl), -S(=O)2N(C 1-6 alkyl)2, C1-C6 alkyl, C1-C6 haloalkyl, C 3-8 Cycloalkyl, C1-C6 alkoxy, C 1-6 Fluorinated alkoxy, 4-7 membered heterocycloalkyl, phenyl, 5- to 6-membered heteroaryl, phenyloxy, C 1-6 Alkylthio, phenylthio, C1-C6 alkyl sulfoxide, phenyl sulfoxide, C1-C6 alkyl sulfone and phenyl sulfone.

32. The compound of claim 30 or a pharmaceutically acceptable salt thereof, wherein each R 19 are independently halogen.

33. The compound of claim 30 or a pharmaceutically acceptable salt thereof, wherein the C3-C8 cycloalkyl group is cyclobutyl, cyclopentyl, cyclohexyl or spiro[3,3]heptyl.

34. A compound as described in any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, wherein R4 is selected from 35. The compound of claim 34 or a pharmaceutically acceptable salt thereof, wherein R4 is 36. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the haloacetyl group is a monosubstituted haloacetyl group or a disubstituted haloacetyl group.

37. A compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 6, wherein R1 is selected from 38. The compound of claim 37 or a pharmaceutically acceptable salt thereof, wherein R1 is 39. The compound of claim 37 or a pharmaceutically acceptable salt thereof, wherein R1 is 40. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from 41. The compound of claim 1 or 40, or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:

42. A pharmaceutical composition comprising a compound according to any one of claims 1 to 41 and a pharmaceutically acceptable excipient.

43. Use of a compound or salt thereof as described in any one of claims 1 to 41 or a pharmaceutical composition as described in claim 42 in the preparation of a medicament for treating or preventing SARS-CoV-2 infection in a patient in need thereof.

44. Use of a compound or salt thereof according to any one of claims 1 to 41 or a pharmaceutical composition according to claim 42 in the preparation of a medicament for inhibiting the protease of SARS-CoV-2 in vivo.

45. The use of claim 44, wherein the compound binds to a cysteine ​​residue of the protease.

46. ​​The use of claim 44 or 45, wherein the compound binds reversibly or irreversibly to the cysteine ​​residue.

47. The use according to claim 44 or 45, wherein the protease is 3CL protease.

48. The use of claim 44 or 45, wherein the cysteine ​​is cysteine ​​145 of 3CL protease.

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