Novel compound, composition comprising same, and use thereof

Abstract

The present invention provides novel compounds, pharmaceutical compositions comprising the compounds, and uses thereof. The compound provided by the invention has improved metabolic stability, and can be used for preventing and treating various diseases or symptoms, such as inflammatory diseases, proliferative diseases and autoimmune diseases.

Description

Technical Field

[0001] This invention relates to novel IL-17 inhibitors, pharmaceutical compositions comprising said compounds, and their uses. Methods for treating, preventing, and managing inflammatory conditions using said compounds and compositions are also disclosed. Background Technology

[0002] Interleukin-17 (IL-17) is a T-cell-derived pro-inflammatory molecule that stimulates epithelial cells, endothelial cells, and fibroblasts to produce other inflammatory cytokines and chemokines, including IL-6, IL-8, G-CSF, and MCP-1. The IL-17 cytokine family (including IL-17A to IL-17F) promotes the maintenance of adaptive and innate immunity (Gaffen SL. Structure and signaling in the IL-17 receptor family. Nat Rev Immunol. 2009; 9(8):556–567). Dysregulation of IL-17 expression may contribute to inflammatory and autoimmune diseases such as psoriasis, psoriatic arthritis, rheumatoid arthritis, and multiple sclerosis (Li X, Bechara R, Zhao J, et al. IL-17 receptor-based signaling and its implications for disease. Nat Immunol. 2019; 20(12):1594–1602). Therefore, they represent highly promising new targets for the treatment of inflammatory diseases.

[0003] Interleukin-17A (IL-17A) is the most studied member of the IL-17 family. It is a pro-inflammatory cytokine that plays a key role in immune and autoimmune-related diseases, including psoriasis, asthma, psoriatic arthritis, and rheumatoid arthritis. (Tollenaere MAX, Hebsgaard J, Ewald DA, et al. Signalling of multiple interleukin (IL)-17 family cytokines via IL-17 receptor A drives psoriasis-related inflammatory pathways. Br JDermatol. 2021; 185(3):585–594; Ostling J, Geest M, Schofield JPR, et al. IL-17—high asthma with features of a psoriasis immunophenotype. Journal of Allergy and Clinical) Immunology. 2019; 144(5): 1198–1213; Blauvelt A, Chiricozzi A. The immunologic role of IL-17 in psoriasis and psoriatic arthritis pathogenesis. Clin Rev Allergy Immunol. 2018; 55(3): 379–390; Kirkham BW, Kavanaugh A, Reich K. Interleukin-17A: a unique pathway in immune-mediated diseases psoriasis, psoriatic arthritis and rheumatoid arthritis. Immunology. 2014; 141(2): 133–142. IL-17A forms homodimers or heterodimers with IL-17A or IL-17F, and is a major cytokine secreted primarily by Th17 cells.It signals through its membrane-bound receptors, IL-17RA and IL-17RC, to regulate the IL-17A signaling pathway, triggering various inflammatory and immune responses. Therefore, IL-17A has become a major research topic in the treatment of inflammation-related diseases.

[0004] Currently, three approved monoclonal antibodies (mAbs) (secukinumab, ixekizumab, and brodalumab) are being used in clinical practice as IL-17A inhibitors to treat a variety of immune diseases. However, drawbacks such as non-oral administration, poor tissue penetration, lack of blood-brain barrier penetration, and often long half-lives limit the application scope of monoclonal antibodies.

[0005] In this regard, LeoPharma A / S and DiceAlpha, Inc. have developed a series of small-molecule IL-17A inhibitors and began Phase I clinical trials in 2021 and 2022, respectively. Eli Lilly has developed a series of potent imidazo[1,2-b]pyridazine derivatives for the treatment of certain symptoms of psoriasis, rheumatoid arthritis, or multiple sclerosis. In 2021, Eli Lilly initiated a Phase I clinical trial for one of these compounds to investigate IL-17A-related diseases. Unfortunately, the clinical trial was discontinued due to hepatotoxicity.

[0006] Therefore, there is still a need to develop small molecule IL-17A inhibitors with improved performance, low toxicity, and high activity, especially IL-17A / A or IL-17A / F inhibitors. Summary of the Invention

[0007] The inventors of this application have surprisingly discovered a series of compounds that act as IL-17A inhibitors, particularly as IL-17A / A or IL-17A / F inhibitors. Compared to currently available small molecule compounds, these compounds exhibit enhanced metabolic stability and higher or comparable activity compared to large molecules such as monoclonal antibodies.

[0008] In a first aspect, the present invention relates to compounds of formula (I):

[0009]

[0010] in

[0011] Part A is selected from the following group:

[0012] A 5-12 membered heteroaryl group containing one or more nitrogen atoms, wherein the heteroaryl group is optionally substituted by one or more substituents selected independently from the group consisting of: halogen, hydroxyl, cyano, -NH2, C. 1-6 Alkyl, C 1-6 Alkoxy, C3-7 cycloalkyl, C 3-7 Cycloalkoxy, phenyl, phenoxy, 5-6-membered heteroaryl and 4-7-membered heterocyclic alkyl, wherein the 5-6-membered heteroaryl and 4-7-membered heterocyclic alkyl contain one or more heteroatoms each independently selected from O, S and N;

[0013] -NH-C(O)-R3, where R3 is selected from the following group:

[0014] C 3-12 Cycloalkyl and 3-12 membered heterocyclic groups, C 5-12 aryl and 5-12 heteroaryl groups, optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, halogen, -CN, C 1-6 Cyanoalkyl, oxo, -NH2, and -OH; the heterocyclic alkyl and heteroaryl groups contain one or more heteroatoms each independently selected from O, S, and N;

[0015] Optionally substituted methyl groups, wherein one or more substituents are independently selected from: C 1-6 Alkoxy, C 6-12 aryl, and 5-12 membered heteroaryl groups containing one or more heteroatoms each independently selected from O, S, and N, each optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy groups, halogens, -CN, and -OH;

[0016] -OC 3-7 cycloalkyl and -O-CH2-C 3-7 Cycloalkyl, -O-CH2-5-7-membered heteroaryl and benzyl, each benzyl, C 3-7 The cycloalkyl and heteroaryl groups are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 cycloalkyl, halogen, -CN, -NH2 and -OH;

[0017] -NH(C 1-6 alkyl), -NH(C) 3-7 cycloalkyl), -N(C) 1-6alkyl)2 and -N(C 1-6 Alkyl)(C 3-7 cycloalkyl groups, each optionally substituted by one or more substituents selected independently from the group consisting of: C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 cycloalkyl, halogen, -CN, -NH2 and -OH;

[0018] C 1-6 Alkyl, C 6-12 Aryl, 5-12 membered heteroaryl, 4-7 membered heterocyclic, each optionally substituted by one or more substituents independently selected from the group consisting of: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocyclic alkyl, halogen, halogenated C 1-6 Alkyl, -CN, -NH2, -OH, phenyl, -NH(C 1-6 Alkyl group), -(CH2) q -NH(CO)-(C 1-4 alkyl) and -NHS(O)2(C 1-6 Alkyl); q is 0, 1, 2, 3 or 4;

[0019] Alternatively, two substituents located on the same or adjacent ring atoms and the ring atom attached to them together form: substituted or unsubstituted C 5-7 Carbocyclic or substituted or unsubstituted 5-7 membered heterocyclic groups;

[0020] Part B is selected from: C 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more heteroatoms independently selected from N, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens; the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups;

[0021] Part C is selected from the following group:

[0022] * A 4-9 membered heterocyclic group containing one or more heteroatoms selected from O, N, and S, wherein the heterocyclic group is optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene (=CH2), C... 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 carbonyl group, -OC 3-6 cycloalkyl, C 1-6Hydroxyalkyl, halogen, -CN, CF3, -CD3, -NH2, =NH, -OH, -COOH, C 1-6 Alkoxy, -SC 1-6 Alkyl, C 1-3 Carbonyl, oxo, -N(C) 1-6 alkyl)2、-NH(C 1-6 alkyl), -NHC(O)-(C 1-6 alkyl), -NHC(O)-NH(C) 1-6 Alkyl), -NHS(O)2-(C 1-6 Alkyl), -NHS(O)2-(C 3-6 cycloalkyl), -S(O)2-(C 1-6 Alkyl), -S(O)2-(C 3-6 cycloalkyl), -C(O)-C 1-6 Alkyl, 4-7 membered heterocyclic and 5-7 membered heteroaryl;

[0023] The substituent is optionally substituted by one or more substituents, each independently selected from the group consisting of: deuterium, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkyl groups, halogens, -CN, CF3, CD3, oxo groups, -NH2, -COOH, -C(O)OC 1-6 Alkyl, -OH, 5-7 membered heteroaryl, 4-7 membered heterocyclic, C 1-6 Hydroxyalkyl and C 3-6 cycloalkyl;

[0024] It can be formed by two substituents located on the same ring atom or adjacent ring atoms and the ring atom attached to them: C 3-7 Carbon rings, or 3-7 membered heterocycles,

[0025] The C 3-7 The carbocyclic ring or 3-7 membered heterocycle is optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene (=CH2), C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-3 Alkyl, halogen, -CN, CF3, -NH2, -OH or oxo group; optionally substituted amino group, wherein one or more substituents are independently selected from: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C1-6 Hydroxyalkyl, C 1-6 Alkyl groups, halogens, -CN, -NO2, -CF3, -NH2, and -OH; the substituents may optionally be substituted with one or more halogens;

[0026] Optionally substituted amino groups, wherein one or more substituents are independently selected from: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkyl groups, halogens, -CN, -NO2, -CF3, -NH2, and -OH; the substituents may optionally be substituted with one or more halogens;

[0027] R1, R1', and R2' are each independently selected from the following groups: hydrogen, deuterium, and C. 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, deuterated C 1-6 Alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl group, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, halogen, halogenated C 1-6 Alkyl, -CN, -N(C) 1-6 Alkyl groups (-2), CF3, -NH2, and -OH;

[0028] R2 is selected from hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, -NH2, -N(C) 1-6 Alkyl)2、-NH-C 1-6 Alkyl group, -NHC(O)-C 1-6 Alkyl, -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-(4-7 membered heterocyclic group), -NHC(O)-(4-7 membered heteroaryl group), -NHC(O)-O-NH-C 1-6 Alkyl group, -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 1-6 Alkyl), -NHC(O)-N(C) 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 1-6 Alkyl, -NHC(O)-OC 3-7 Cycloalkyl, -NHC(O)-N(C) 1-6 Alkyl)2、-NHC(O)-N(C 3-7 cycloalkyl)2、-NHC(O)-O-CH2-C 3-7cycloalkyl, -NHSO2-C 1-6 Alkyl group, -NHSO2-NH-C 1-6 Alkyl groups, and -NHSO2-N(C 1-6 Alkyl)2,

[0029] Optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 3-7 Cycloalkyl, deuterium, halogen, -CN, CF3, -NH2, and -OH; the heterocyclic or heteroaryl group contains one or more heteroatoms each independently selected from O, S, and N;

[0030] R a and R a Each is independently selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne, halogen, -CN, CF3, -NH2, -OH, -C 1-3 Alkoxy, C 3-7 cycloalkyl and C 3-7 Heterocyclic groups;

[0031] R b and R b Each is independently selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 3-7 cycloalkyl, C 5-10 Aryl, and C 5-10 The heteroaryl group is optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-7 Cycloalkyl, halogen, -CN, CF3, -NH2, -OH, and -C 1-3 alkoxy; or R b and R b 'Together with the carbon atoms they are bonded to form: C 3-8 Carbon ring, C 5-9 Spiral ring, C 5-10 Aromatic rings or C 5-10 Aromatic heterocyclic ring, optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene, C... 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -C 1-3Alkoxy, halogen, -CN, CF3, -NH2, and -OH; the substituents may optionally be substituted with one or more halogens;

[0032] L does not exist, or is selected from -O- and -NH-;

[0033] m is 0, 1, or 2; and

[0034] n is 0 or 1;

[0035] Or its enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof.

[0036] In some preferred embodiments, fragment C is connected to the rest of the general formula via the N atom in fragment C.

[0037] In some embodiments, portion B is selected from C6 aryl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more heteroatoms each independently selected from N, wherein the C6 aryl or 6-membered heteroaryl group is optionally substituted with one or more halogens; wherein the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo-substituted groups.

[0038] In some implementations, L is absent, m is 0, and n is 1, and the compound is as shown in formula (Ia):

[0039]

[0040] in

[0041] Part A1 is as defined in Part A;

[0042] Part B1 is as defined in part B;

[0043] Part C1 is as defined in part C;

[0044] R b1 For example, R b Defined;

[0045] R b1' For example, R b’ Defined; and

[0046] R1, R 1' R 2' And R2 as described above.

[0047] In some embodiments, a portion A1 is selected from the group consisting of 5-12-membered heteroaryl groups containing one or more N atoms, wherein the heteroaryl group is optionally substituted by one or more substituents independently selected from the group consisting of halogens, hydroxyl groups, cyano groups, -NH2 groups, C4 groups, etc. 1-6 Alkyl, C 1-6 Alkoxy, C3-7 Cycloalkyl groups, and 4-7 membered heterocycloalkyl groups containing one or more heteroatoms each independently selected from O, S and N.

[0048] In some implementations, part of B1 is selected from C. 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group may optionally be substituted with one or more halogens, and the 6-membered heterocyclic group may optionally be substituted with one or more halogens or oxo groups.

[0049] In some embodiments, a portion of C1 is selected from 4-9 membered heterocyclic groups containing one or more N atoms, said heterocyclic group optionally being substituted by one or more substituents independently selected from the group consisting of halogens, methylene, and C. 1-6 Alkyl groups, wherein the substituents are optionally substituted with one or more halogens, or two substituents on the same ring atom together with the ring atoms to which they are attached form a C12 group. 3-7 Carbon ring.

[0050] In some implementations, R1, R1', and R2' are each independently selected from the group consisting of hydrogen, CF3, and C. 1-6 alkyl.

[0051] In some implementations, R2 is selected from -NH2, -NHC(O)-C 1-6 Alkyl group, -NHC(O)-NH-(C 1-6 alkyl), -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-N(C 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 3-7 The cycloalkyl group, and -NHC(O)-(4-7-membered heterocyclic group) and -NHC(O)-(4-7-membered heteroaryl group), are each optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl and C 1-6 Alkoxy group; the heterocyclic group contains one or more heteroatoms, each independently selected from O, S and N.

[0052] In some implementation schemes, R b1 and R b1 Each is independently selected from one or more Cs 3-7 Cycloalkyl-substituted C 3-7 cycloalkyl or C 2-6 alkenyl; or R b1 and R b1'Formed together with the carbon atoms they are attached to: optionally by one or more C atoms.' 1-6 alkyl or halogen-substituted C 3-8 Carbon ring.

[0053] In some preferred embodiments, R2' is CF3 or C 1-3 Alkyl group; and / or part of B1 selected from phenyl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N atoms, wherein the phenyl group is substituted with two halogens, the 6-membered heteroaryl group is unsubstituted or substituted with halogens, and the 6-membered heterocyclic group is unsubstituted or substituted with halogens or oxo groups; and / or part of C1 selected from 5-9-membered heterocyclic groups containing one or more N atoms, wherein the heterocyclic group is substituted with =CF2, and optionally substituted with one or more halogens or C 1-6 Alkyl substitution, or two substituents located on the same ring atom together with the ring atom to which they are attached, form C. 3-7 Carbon rings; and / or R b1 and R b1 Together with the carbon atoms they are attached to, they form C atoms that are substituted with =CF2. 5-8 Carbon ring.

[0054] In some implementations, L is absent, m is 0, n is 1, and the compound is as shown in formula (Ib):

[0055]

[0056] in

[0057] Part D is selected from the following group: C 1-6 Alkyl, C 3-7 cycloalkyl, -OC 3-7 cycloalkyl, -O-CH2-C 3-7 Cycloalkyl, -O-CH2-5-7-membered heteroaryl, -NH(C 1-6 alkyl), -NH(C) 3-7 cyclo group), -N(C) 1-6 Alkyl)2, -N(C 1-6 Alkyl)(C 3-7 cycloalkyl), C 6-12 Aryl, 5-12 membered heteroaryl, 4-7 membered heterocyclic,

[0058] Optionally substituted by one or more substituents independently selected from the following group: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocyclic alkyl, halogen, halogenated C 1-6 Alkyl, C 1-6Cyanoalkyl, -oxo, -CN, -NH2, -OH, phenyl, -NH(C 1-6 Alkyl group), -(CH2) q -NH(CO)-(C 1-4 Alkyl groups), and -NHS(O)2(C 1-6 Alkyl; q is 0, 1, 2, 3, or 4; or, two substituents located on the same or adjacent ring atoms, together with the ring atoms they are attached to, form a substituted or unsubstituted C. 5-7 A carbocyclic group, or a substituted or unsubstituted 5-7 membered heterocyclic group; the heterocyclic alkyl, heterocyclic and heteroaryl groups contain one or more heteroatoms independently selected from O, S and N;

[0059] R c and R c Each is independently selected from hydrogen, deuterium, and C 1-3 Alkyl, C 1-3 Alkyl groups, halogens, -CN, -CF3, -CD3, -NH2, and -OH;

[0060] p is 0 or 1;

[0061] Part B2 is as defined in part B;

[0062] Part C2 is as defined in part C;

[0063] R b2 For example, R b Defined;

[0064] R b2' For example, R b' Defined; and

[0065] R1, R 1' R 2' And R2 as described above.

[0066] In some preferred embodiments, a portion of D is selected from C. 3-6 Cycloalkyl, 5-6 membered heterocyclic, and 5-6 membered heteroaryl containing one or more heteroatoms selected from N, O, and S; optionally substituted with one or more substituents selected from the group consisting of: halogen, α-oxo, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Cyanoalkyl, and -CN.

[0067] In some preferred embodiments, a portion of B2 is selected from C. 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens; the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups.

[0068] In some preferred embodiments, p is o.

[0069] In some preferred embodiments, a portion of C2 is selected from the group consisting of:

[0070] The amino group, optionally substituted with one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, halogen or halogenated C 1-6 alkyl;

[0071] A 4-9 membered heterocyclic group containing one or more heteroatoms selected from N, O, and S, wherein the heterocyclic group is optionally substituted by one or more substituents independently selected from the group consisting of methylene, C, and S. 1-6 Alkyl, C 2-6 alkenyl, C 1-6 Alkoxy, -SC 1-6 Alkyl, C 3-8 carbonyl group, -OC 3-6 Cycloalkyl, -NH2, CF3, CD3, CN, -oxo, =NH, -N(C 1-6 alkyl)2、-NH(C 1-6 alkyl), -NHC(O)-(C 1-6 alkyl), -NHC(O)-NH(C) 1-6 Alkyl), -NHS(O)2-(C 1-6 Alkyl), -NHS(O)2-(C 3-6 cycloalkyl), -S(O)2-(C 1-6 Alkyl), -S(O)2-(C 3-6 cycloalkyl), C 1-6 Hydroxyalkyl, -C(O)-C 1-6 Alkyl, halogen, -OH, -COOH, 5-7 membered heterocyclic groups, and 5-7 membered heteroaryl groups;

[0072] The substituents are optionally replaced by one or more halogens, deuterium, CN, CF3, -oxo, CD3, -OH, -COOH, -C(O)OC. 1-6 Alkyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, 5-7 membered heteroaryl, 4-7 membered heterocyclic and C 3-6 Cycloalkyl substitution;

[0073] Optionally, two substituents located on the same or adjacent ring atoms, together with the ring atoms they are attached to, form: C 3-7A carbon ring, or a 3-7 membered heterocycle containing one or more heteroatoms selected from N, O and S, or a 5-7 membered heteroaryl;

[0074] The C 3-7 The carbocyclic ring or 3-7 membered heterocycle is optionally substituted by one or more substituents independently selected from the group consisting of: methylene (=CH2), halogen, -oxo, C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-3 Alkoxy, -CN, -CF3, -NH2, and -OH.

[0075] In some preferred embodiments, R2' is H, and / or R2 is selected from -NH2, -NHC(O)-OC 1-6 Alkyl group, -NHC(O)-C 1-6 Alkyl group, -NHC(O)-NH-(C 1-6 alk group), -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-N(C 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 3-7 Cycloalkyl, -NHC(O)-O-CH2-C 3-7 Cycloalkyl, -NHC(O)- (4-7 membered heterocyclic groups), -NHC(O)-N(C 1-6 Alkyl)2、-NHC(O)-N(C 3-7 Cycloalkyl)2 and -NHC(O)- (4-7-membered heteroaryl), each optionally substituted by one or more substituents independently selected from the group consisting of: deuterium, halogen, C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 1-6 Alkoxy and C 3-7 cycloalkyl;

[0076] The heterocyclic or heteroaryl group comprises one or more heteroatoms independently selected from O, S, and N.

[0077] In some preferred embodiments, R b2 and R b2 Each independently selected from C 3-7 cycloalkyl; or R b2 and R b2 Together with the carbon atoms they are attached to, they form: C 3-8The carbocyclic ring, or a 5-7 membered heterocycle, is optionally substituted by one or more substituents independently selected from the group consisting of: methylene, C... 2-6 alkenyl, C 1-6 Alkyl or halogen; the substituent may optionally be replaced by one or more halogens;

[0078] Alternatively, two substituents located on the same or adjacent ring atoms, together with the ring atoms they are attached to, form: C 3-6 A carbocyclic ring, or a 4-7 membered heterocyclic group; wherein the carbocyclic ring and heterocyclic group are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 2-6 alkenyl, C 1-6 Alkyl or halogen; the substituent may optionally be replaced by one or more halogens.

[0079] In some preferred embodiments, R b2 and R b2 Each independently selected from C 3-5 cycloalkyl; or R b2 and R b2 Together with the carbon atoms they are attached to, they form C 3-7 The carbocyclic ring or 5-7 membered heterocycle is optionally substituted by one or more substituents independently selected from the group consisting of: methylene (=CH2), C 2-6 alkenyl, C 1-6 Alkyl or halogen; the substituent may optionally be replaced by one or more halogens.

[0080] In some preferred embodiments, part C2 is a 4-9 membered heterocyclic group containing at least one N atom and no or more heteroatoms selected from O, N and S.

[0081] In some preferred embodiments, part C2 is connected to the remainder of formula Ib via N atoms in part C2.

[0082] In some preferred embodiments, R2' is CF3 or C 1-3 Alkyl group; and / or part of B2 selected from phenyl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N atoms, wherein the phenyl group is substituted with two halogens, the 6-membered heteroaryl group is unsubstituted or substituted with one halogen, and the 6-membered heterocyclic group is unsubstituted or substituted with one halogen or oxo group, and / or part of C2 selected from 5-9-membered heterocyclic groups containing one or more N atoms, wherein the heterocyclic group is substituted with a =CF2 group, and optionally substituted with one or more halogens, or C 1-6 Alkyl substitution, or two substituents located on the same or adjacent ring atoms forming a C12 ring with the ring atoms to which they are attached. 3-7 Carbon rings; and / or R b2 and Rb2 Together with the carbon atoms they are attached to, they form C atoms that are substituted with a =CF2. 5-8 Carbon ring.

[0083] In some preferred embodiments, a portion of C2 is selected from the group consisting of a 6-membered heterocyclic group containing at least one N atom, wherein the heterocyclic group is optionally substituted by one or more substituents independently selected from the group consisting of: halogen, -COOH, -CN, -NH2, -NH-CO-NH(C 1-6 alkyl), -N(C) 1-6 alkyl)2、-NH(C 1-6 Alkyl), -NH-S(O)2(C 1-6 Alkyl), -S(O)2(C 1-6 Alkyl group, =CH2, =CF2, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 2-6 Alkenyl, halogenated =CH2, halogenated C 2-6 Alkenyl, 4-7 membered heterocyclic groups, and C 3-8 Cycloalkyl; each substituent may be optionally substituted by one or more substituents selected from the group consisting of: D, halogen, -CN, -OH, -COOH, C. 1-6 Alkyl, C 1-6 Hydroxyalkyl, halogenated C 1-6 Alkyl groups, 4-6-membered heterocyclic groups containing one or more heteroatoms selected from N, O and S, and 6-membered heteroaryl groups containing one N atom;

[0084] Alternatively, two substituents located on adjacent atoms or on the same atom, together with the atoms they are attached to, form: 4-7 membered heterocycles, C 3-6 The carbon ring, or 5-7 membered heteroaromatic ring, may be optionally halogenated, β-oxo-substituted, or C-substituted. 1-6 Alkyl, =CF2, and -CF3 substitutions.

[0085] In some preferred embodiments, the C2 portion is selected from the group consisting of a 6-membered heterocyclic group containing at least one N atom, the heterocyclic group optionally being substituted by one or more substituents independently selected from the group consisting of -CH3 or =CF2; the 6-membered heterocyclic group being connected to the remainder of the general formula via the N atom.

[0086] In some preferred embodiments, the compound is as shown in formula Ib-1.

[0087]

[0088] Among them, R1, R1', R2', R2, R b2 R b2 '、Rc and R c ', p, part B2 and part D are as described above.

[0089] In some preferred embodiments, R b2 and R b2 Each independently selected from C 3-7 cycloalkyl; or, R b2 and R b2 Together with the carbon atoms they are attached to, they form: C 3-8 The carbon ring; preferably, it forms cycloheptane or cyclohexane, unsubstituted or substituted with one =CF2.

[0090] In some preferred embodiments, R b2 and R b2 Each is independently selected from cyclopropyl; or R b2 and R b2 Together with the carbon atoms they are attached to, they form cycloheptane or cyclohexane substituted with =CF2.

[0091] In some preferred embodiments, a portion of B2 is a halophenyl group.

[0092] In some preferred embodiments, part D is a 5-6 membered heteroaryl group, optionally substituted by one or more substituents selected from the group consisting of: C 1-6 Alkyl groups and halogens.

[0093] In some preferred embodiments, C2 is a 6-membered heterocyclic group containing at least one N atom, substituted with a =CF2 group; and / or R b2 and R b2 'Together with the carbon atoms they are attached to, they form: C 5-8 The carbon ring; preferably, it forms cycloheptane or cyclohexane, unsubstituted or substituted with one =CF2.

[0094] In some preferred embodiments, the compound is as shown in formula Ib-2.

[0095]

[0096] Among them, R1, R1', R2', R2, R c and R c ', p, part B2 and part D are as described above.

[0097] In some implementations, L is absent, m is 0, n is 0, and the compound is as shown in formula (Ic):

[0098]

[0099] in

[0100] R b3 For example, R b Defined;

[0101] R b3 'Like R b’ Defined;

[0102] Part B3 is as defined in part B;

[0103] Part C3 is as defined in part C;

[0104] p is 0 or 1;

[0105] Part of D1 is selected from the following group: C 6-12 aryl, 5-12 heteroaryl, optionally substituted by one or more substituents independently selected from the group consisting of: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, -CN, -NH2, and -OH; the heteroaryl group comprises one or more heteroatoms independently selected from O, S, and N;

[0106] R2 and R2' are independently selected from the following group: hydrogen, CF3, and C. 1-6 alkyl;

[0107] R c and R c Each is independently selected from hydrogen, deuterium, and C 1-3 Alkyl, C 1-3 Alkyl groups, halogens, -CN, -CF3, -CD3, -NH2, and -OH;

[0108] Or its enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof.

[0109] In some preferred embodiments, a portion of B3 is selected from C. 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens, wherein the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups.

[0110] In some preferred embodiments, part C3 is selected from the group consisting of: optionally one or more C 1-6 Alkyl-substituted amino groups, wherein the C 1-6 Alkyl groups may be optionally substituted by one or more substituents selected from the group consisting of halogens and halogenated C groups. 1-6Alkyl group; a 4-9 membered heterocyclic group containing one or more nitrogen atoms, said heterocyclic group optionally substituted with one or more substituents selected from the group consisting of: halogen, methylene, and C. 2-6 alkenyl, the methylene and C 2-6 The alkenyl group may be optionally substituted by one or more substituents selected from the group consisting of: halogen, C 1-6 Alkyl groups, or two substituents located on the same ring atom, together with the ring atoms to which they are attached, form C14. 3-7 Carbon ring.

[0111] In some preferred embodiments, R b3 and R b3 Each is independently selected from hydrogen and C 3-7 cycloalkyl or with one or more C 3-7 Cycloalkyl-substituted C 2-6 alkenyl; or R b3 and R b3' Together with the carbon atoms they are attached to, they form: C atoms optionally substituted with one or more substituents selected from the group consisting of: 3-8 Carbocyclic ring: methylene, C 1-6 Alkyl, C 2-6 Alkenyl and halogen; the substituents may optionally be replaced by one or more halogens.

[0112] In some preferred embodiments, a portion of D1 is selected from the group consisting of phenyl and 5-6-membered heteroaryl groups, optionally substituted by one or more substituents independently selected from the group consisting of C 1-6 Alkyl, C 1-6 Alkoxy, halogen, and -CN; the heteroaryl group comprises one or more heteroatoms independently selected from O, S, and N;

[0113] Part of B3 is selected from phenyl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N heteroatoms, wherein the phenyl or 6-membered heteroaryl group is optionally substituted with one or more halogens, and the 6-membered heterocyclic group is optionally substituted with one or more substituents selected from the group consisting of halogens and halogenated C. 1-6 alkyl;

[0114] Some C3s are selected from the following group: -N(C 1-6 Alkyl group 2, optionally oxidized by one or more halogens or halogenated C 1-6 Alkyl substitution; a 4-9 membered heterocyclic group containing one or more N atoms, said heterocyclic group optionally substituted with one or more halogens, methylene groups, or C atoms. 2-6 Alkenyl substitution, the methylene and C 2-6 Alkenyl groups are optionally converted to one or more halogens or C. 1-6 Alkyl substitution; the 4-9 membered heterocyclic groups are connected to the rest of the structure via N atoms.

[0115] In some preferred embodiments, portion B3 is selected from phenyl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N atoms, wherein the phenyl is substituted with two halogens, the 6-membered heteroaryl group is unsubstituted or substituted with one halogen, and the 6-membered heterocyclic group is unsubstituted or substituted with one halogen or oxo group; and / or, portion C3 is selected from 5-9-membered heterocyclic groups containing one or more N atoms, wherein the heterocyclic group is substituted with a =CF2 and optionally substituted with one or more halogens or C 1-6 Alkyl substitution, or two substituents on the same ring atom and the ring atom attached to them together to form C 3-7 Carbon rings; and / or, R b3 and R b3 Together with the carbon atoms they are attached to, they form C atoms that are substituted with a =CF2. 5-8 Carbon ring.

[0116] In another preferred embodiment, the heterocyclic group and carbocyclic group may be saturated or partially unsaturated, or unsaturated, substituted or unsubstituted, aromatic or non-aromatic.

[0117] In some preferred embodiments, the heterocyclic group and the carbocyclic group can be fused, bridged, or spirally connected.

[0118] In another preferred embodiment, each group is a corresponding group in a specific compound in the examples.

[0119] In some preferred embodiments, the compound is selected from the group consisting of:

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126]

[0127]

[0128]

[0129]

[0130] Or its enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof.

[0131] It should be understood that the compounds of the present invention are not limited to those listed above.

[0132] In a second aspect, the present invention relates to a pharmaceutical composition comprising the compound of the present invention and a pharmaceutically acceptable excipient.

[0133] In some embodiments, the pharmaceutical composition can be used to prepare a single, unit dosage form. In some embodiments, the single-unit dosage forms provided by the present invention are suitable for oral, mucosal, parenteral, topical, transdermal, or transdermal administration to a patient. Examples of the dosage forms include, but are not limited to: tablets; capsules; capsules, such as soft elastic gelatin capsules; flat capsules; lozenges; dispersants; suppositories; powders; aerosols; gels; liquid dosage forms suitable for oral or mucosal administration to a patient, including suspensions, solutions, and elixirs; liquid dosage forms suitable for parenteral administration to a patient; eye drops or other ophthalmic preparations suitable for topical administration; and sterile solids that can be reconstituted into liquid dosage forms suitable for parenteral administration to a patient. In some embodiments, the dosage form comprises 1 to about 1000 mg, about 5 to about 500 mg, about 10 to about 350 mg, or about 50 to about 200 mg of the compound provided herein or its enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts. The specific amounts of the compounds, their enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts provided herein will depend on the particular drug used, the disease or condition being treated or prevented.

[0134] In some embodiments, the pharmaceutical composition comprises the compound provided herein or its enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof, and optionally a second active agent for the treatment of inflammatory diseases, proliferative diseases and autoimmune diseases.

[0135] In a third aspect, this application relates to a method of treating a disease or condition in a subject in need, wherein the method comprises administering to the subject a therapeutically effective amount of the compound of this application, or its enantiomer, stereoisomer, solvate or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of this application, wherein the disease or condition is selected from the group consisting of inflammatory diseases, proliferative diseases and autoimmune diseases.

[0136] In some embodiments, the disease or condition is selected from one or more of the following: plaque psoriasis, guttate psoriasis, reversal psoriasis, pustular psoriasis, erythrodermic psoriasis, psoriatic arthritis, ankylosing spondylitis, hidradenitis suppurativa, palmoplantar psoriasis, airway inflammation, ankylosing spondylitis, asthma, rheumatoid arthritis, osteoarthritis, spondyloarthritis, bone erosion, intraperitoneal abscess and adhesions, IBD (inflammatory bowel disease), Crohn's disease, allograft rejection, psoriasis, psoriatic arthritis, certain types Cancer, angiogenesis, atherosclerosis and multiple sclerosis, erythema, reactions to allergen exposure, Helicobacter pylori-associated gastritis, bronchial asthma, asthma, allogeneic graft rejection (e.g., kidney), systemic lupus erythematosus, lupus nephritis, Behcet's disease, ulcerative colitis, rheumatoid arthritis (RA), inflammatory bowel disease, Wegener's granulomatosis, sarcoidosis, systemic sclerosis, insulin-dependent diabetes mellitus, septic shock syndrome, Alzheimer's disease, inflammatory eye disease, uveitis and non-infectious uveitis.

[0137] In a fourth aspect, this application relates to the use of compounds of the present invention, or their enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof, or pharmaceutical compositions of the present invention in the preparation of medicaments for the prevention or treatment of diseases or conditions, wherein said diseases or conditions are selected from the group consisting of inflammatory diseases, proliferative diseases, autoimmune diseases, or combinations thereof.

[0138] In some embodiments, the disease or condition is selected from one or more of the following: plaque psoriasis, guttate psoriasis, reversal psoriasis, pustular psoriasis, erythrodermic psoriasis, psoriatic arthritis, ankylosing spondylitis, hidradenitis suppurativa, palmoplantar psoriasis, airway inflammation, ankylosing spondylitis, asthma, rheumatoid arthritis, osteoarthritis, spondyloarthritis, bone erosion, intraperitoneal abscess and adhesions, IBD (inflammatory bowel disease), Crohn's disease, allograft rejection, psoriasis, psoriatic arthritis, certain types Cancer, angiogenesis, atherosclerosis and multiple sclerosis, erythema, reactions to allergen exposure, Helicobacter pylori-associated gastritis, bronchial asthma, asthma, allogeneic graft rejection (e.g., kidney), systemic lupus erythematosus, lupus nephritis, Behcet's disease, ulcerative colitis, rheumatoid arthritis (RA), inflammatory bowel disease, Wegener's granulomatosis, sarcoidosis, systemic sclerosis, insulin-dependent diabetes mellitus, septic shock syndrome, Alzheimer's disease, inflammatory eye disease, uveitis and non-infectious uveitis.

[0139] In a fifth aspect, this application relates to compounds of the invention, or their enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof, for methods of preventing or treating diseases or conditions, or pharmaceutical compositions of the invention, wherein the disease or condition is selected from inflammatory diseases, proliferative diseases and autoimmune diseases.

[0140] In some embodiments, the disease or condition is selected from one or more of the following: plaque psoriasis, guttate psoriasis, reversal psoriasis, pustular psoriasis, erythrodermic psoriasis, psoriatic arthritis, ankylosing spondylitis, hidradenitis suppurativa, palmoplantar psoriasis, airway inflammation, ankylosing spondylitis, asthma, rheumatoid arthritis, osteoarthritis, spondyloarthritis, bone erosion, intraperitoneal abscess and adhesions, IBD (inflammatory bowel disease), Crohn's disease, allograft rejection, psoriasis, psoriatic arthritis, certain types Cancer, angiogenesis, atherosclerosis and multiple sclerosis, erythema, reactions to allergen exposure, Helicobacter pylori-associated gastritis, bronchial asthma, asthma, allogeneic graft rejection (e.g., kidney), systemic lupus erythematosus, lupus nephritis, Behcet's disease, ulcerative colitis, rheumatoid arthritis (RA), inflammatory bowel disease, Wegener's granulomatosis, sarcoidosis, systemic sclerosis, insulin-dependent diabetes mellitus, septic shock syndrome, Alzheimer's disease, inflammatory eye disease, uveitis and non-infectious uveitis.

[0141] In some embodiments, the compounds provided by this invention, or their enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, are administered in combination with another drug (“second active agent”) or treatment. The second active agent includes small molecules and large molecules (e.g., proteins and antibodies). Other therapies that can be used in combination with the compounds provided by this invention include, but are not limited to, surgery, immunotherapy, biotherapy, radiotherapy, and other non-pharmacological therapies that can be used to treat or prevent the various diseases described herein.

[0142] Brief description of the attached figures

[0143] Figure 1A -J shows a comparison of the efficacy of the tested compound with the approved anti-IL-17A antibody secukinumab in inhibiting IL-17A / A activation. Figure 1A The efficacy of secukinumab; Figure 1B-1J The therapeutic effects of compounds in representative examples.

[0144] Figure 2A -H shows the comparative efficacy of the test compound with the approved anti-IL-17A antibody secukinumab in inhibiting IL-17A / F activation. Figure 2A The efficacy of secukinumab; Figure 2B-2H The therapeutic effects of compounds in representative examples. Detailed Implementation

[0145] definition

[0146] As used herein, unless otherwise stated, the term "alkyl" refers to a saturated straight-chain or branched hydrocarbon having the number of carbon atoms described herein. Representative saturated straight-chain alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl; while saturated branched alkyl groups include isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, etc. The term "alkyl" also includes cycloalkyl. In some embodiments, the alkenyl group is optionally substituted as described elsewhere herein.

[0147] As used herein, unless otherwise stated, alkenyl refers to a straight-chain or branched hydrocarbon having the number of carbon atoms as described herein and containing one or more double bonds. Exemplary alkenyl carbon chains contain 2 to 20 carbons and 1 to 8 double bonds. Examples of alkenyl include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, allyl, butenyl, and 4-methylbutenyl. In some embodiments, the alkenyl group is optionally substituted as described elsewhere herein.

[0148] As used herein, unless otherwise stated, alkenyl means a straight-chain or branched hydrocarbon having the number of carbon atoms as described herein and containing one or more triple bonds. Exemplary alkynyl carbon chains contain 2 to 20 carbons and 1 to 8 triple bonds. Exemplary alkynyl groups in this document include, but are not limited to, ethynyl, propynyl, and butynyl. As used herein, lower alkyl, lower alkenyl, and lower alkynyl groups refer to carbon chains having about 1 or about 2 carbons to about 6 carbons.

[0149] As used herein, unless otherwise specified, the term "carbocyclic" or "carbocyclic" refers to a non-aromatic ring or ring system containing only carbon atoms in its ring skeleton. When a carbocyclic group is a ring system, two or more rings may be connected together by fusion, bridging, or helical linkage. A carbocyclic group can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. Therefore, carbocyclic groups include cycloalkyl, cycloalkenyl, and cycloynyl groups. A carbocyclic group can have 3 to 20 carbon atoms, although this definition also covers instances of the term "carbocyclic" where no numerical range is specified. A carbocyclic group can also be a medium-sized carbocyclic group having 3 to 10 carbon atoms. A carbocyclic group can also be a carbocyclic group having 3 to 6 carbon atoms. A carbocyclic group can be specified as "C". 3-6 "Carbocyclic" or similar names. Examples of carbocyclic compounds include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, adamantyl, and spiro[2.5]octyl.

[0150] As used herein, unless otherwise stated, the term "cycloalkyl" refers to an alkyl group that is cyclic and contains 3 to 9, 3 to 6, or 3 to 5 carbon atoms, without alternating or resonant double bonds between the carbon atoms. It may contain 1 to 4 rings. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cyclooctyl. A cycloalkyl group may be substituted with one or more substituents. In some embodiments, the cycloalkyl group may be a cycloalkyl group fused with an aryl or heteroaryl group. Bridged ring systems are also included in the definition of "cycloalkyl".

[0151] As used herein, unless otherwise stated, the term "heterocyclic alkyl" or "heterocyclic group" refers to a cycloalkyl group in which one or more carbon atoms are substituted with heteroatoms (e.g., but not limited to N, S, and O). In some embodiments, the heterocyclic alkyl contains 2 to 8, 2 to 7, 2 to 5, or 2 to 4 carbon atoms. Heterocyclic alkyl groups can be attached to the host structure at heteroatoms or carbon atoms to produce stable compounds. Exemplary heterocyclic alkyl or heterocyclic groups include, but are not limited to, morpholino, thiomorpholino, pyrano, imidazoalkyl, oxazolyl, pyrazolyl, pyrrolidinyl, pyrrolino, tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydrothiazolyl, piperidinyl, azacyclic butyl, oxacyclic butyl, and piperazine.

[0152] As used herein, unless otherwise stated, the term "aromatic" refers to a ring or ring system having a conjugated π-electron system, including carbocyclic aromatic groups (such as phenyl) and heterocyclic aromatic groups (such as pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings sharing adjacent atomic pairs) groups, provided that the entire ring system is aromatic.

[0153] As used herein, unless otherwise stated, the term "aryl" refers to a carbocyclic aromatic ring containing 5 to 10 ring atoms. All ring atoms in a carbocyclic aryl group are carbon atoms. Aromatic ring structures include compounds having one or more ring structures, such as monocyclic and bicyclic compounds. Exemplary aryl groups include phenyl and naphthyl.

[0154] As used herein, unless otherwise stated, "heteroaryl" refers to a monocyclic or polycyclic aromatic ring system in which, in some embodiments, in a 5- to 10-membered aromatic ring system, one or more atoms in the ring system (1 to 3 atoms in one embodiment) are heteroatoms (i.e., non-carbon elements) including, but not limited to, nitrogen, oxygen, or sulfur. Examples of monocyclic heteroaryl groups include, but are not limited to: furanyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyrroleyl, thiadiazolyl, thiazolyl, thiophene, tetrazolyl, triazinyl, and triazolyl. Examples of bicyclic heteroaryl groups include, but are not limited to: benzofuranyl, benzimidazolyl, benzoisoxazolyl, benzopyranyl, benzothiadiazolyl, benzothiazolyl, benzothiophenyl, benzotriazolyl, benzoxazolyl, furanopyridyl, imidazopyridyl, imidazothiazolyl, indoleazinyl, indoleyl, indoleyl, isobenzofuranyl, isobenzothiaphenyl, isoindoleyl, isoquinolinyl, isothiazolyl, naphridinyl, oxazolopyridyl, phthalazinyl, pteridinyl, purine, pyridopyridyl, pyrrolopyridyl, quinolinyl, quinoxalolinyl, quinazolinyl, thiadiazopyrimidinyl, and thienzopyridyl. In some embodiments, the heteroaryl group is optionally substituted with one or more substituents as defined elsewhere herein.

[0155] As used herein, unless otherwise stated, the term "alkoxy" or "alkyloxy" refers to a stable straight-chain or branched or cyclic hydrocarbon group, or a combination thereof, consisting of the stated number of carbon atoms and one or more (in one embodiment, 1-3) O atoms, wherein at least one O atom is located at the position where the alkoxy or alkyloxy group is attached to the remainder of the molecule. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, and tert-butoxy. In one embodiment, the alkoxy group is optionally substituted as described elsewhere herein.

[0156] As used herein, unless otherwise stated, the term "halogen" means F, CI, Br, or I. A "halogenated" group means that one or more hydrogen atoms in the group have been replaced by a halogen.

[0157] As used herein, unless otherwise stated, the term "methylene" refers to =CH2.

[0158] As used herein, unless otherwise stated, the term “oxo” is represented by (=O) as an alternative to other common representations.

[0159] As used herein, unless otherwise stated, the term "deuterium" refers to an isotope of hydrogen with one proton and one neutron in its nucleus, and has twice the mass of ordinary hydrogen, usually denoted by "D". A group that is "deuterated" means that one or more hydrogen atoms in the group are replaced by deuterium (D).

[0160] If no number of any given substituents is specified, one or more substituents may be present.

[0161] As used herein, unless otherwise stated, the term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic acid (including inorganic and organic acids). Suitable non-toxic acids include inorganic and organic acids, such as, but not limited to, hydrobromic acid, hydrochloric acid, acetic acid, anthranilic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, vinylsulfonic acid, formic acid, fumaric acid, furoic acid, gluconic acid, glutamic acid, glucuronic acid, galacturonic acid, glycine, hydroxyethanesulfonic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucoic acid, nitric acid, dihydroxynaphthyl acid, pantothenic acid, phenylacetic acid, propionic acid, phosphoric acid, salicylic acid, stearic acid, succinic acid, sulfanilic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc.

[0162] As used herein, and unless otherwise stated, the term "solvent" refers to a compound that further comprises stoichiometric or nonstoichiometric amounts of solvent bound together by noncovalent intermolecular forces. When the solvent is water, the solvate is a hydrate.

[0163] As used herein, unless otherwise stated, the term "stereoisomer" includes all stereoisomerically pure and stereoisomerically enriched compounds provided herein.

[0164] As used herein, unless otherwise stated, the term "enantiomer" includes all enantiomerically pure and enantiomerically enriched compounds provided herein.

[0165] As used herein, and unless otherwise stated, the term "stereoisomeric pure" means a composition comprising one stereoisomer of a compound and substantially free of other stereoisomers of that compound. For example, a stereopure composition of a compound having one chiral center will substantially free of the opposite enantiomers of that compound. A stereopure composition of a compound having two chiral centers will substantially free of other diastereomers of that compound. A typical stereoisomeric pure compound comprises: more than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of another stereoisomer of the compound; more than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of another stereoisomer of the compound; more than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of another stereoisomer of the compound; more than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of another stereoisomer of the compound; more than about 98% by weight of one stereoisomer of the compound and less than about 2% by weight of another stereoisomer of the compound; or more than about 99% by weight of one stereoisomer of the compound and less than about 1% by weight of another stereoisomer of the compound.

[0166] As used herein, unless otherwise stated, the term "enantiomerically pure" refers to a stereoisomerically pure composition of compounds having a single chiral center. Similarly, the term "enantiomerically enriched" refers to a stereoisomerically enriched composition of compounds having a single chiral center.

[0167] As used herein, unless otherwise stated, the prefixes R and S are used to indicate the absolute configuration of a molecule around its chiral center.

[0168] As used herein, and unless otherwise stated, the terms “about” or “approximately” refer to an acceptable error for a particular value as determined by one of ordinary skill in the art, depending in part on how the value was measured or determined. In some embodiments, the terms “about” or “approximately” mean within 1, 2, 3, or 4 standard deviations. In some embodiments, the terms “about” or “approximately” mean within 50%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.05% of a given value or range.

[0169] As used herein, unless otherwise stated, the term "pharmaceuticalally acceptable excipient" means a pharmaceutically acceptable material, composition, or carrier, such as a liquid or solid filler, diluent, solvent, or encapsulating material. Each carrier must be "acceptable," meaning it is compatible with the other components of the formulation and is harmless to the patient. Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars, such as lactose, glucose, and sucrose; starches, such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; tragacanth gum powder; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols, such as propylene glycol; polyols, such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffers, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; phosphate buffer solutions; and other non-toxic and compatible substances used in pharmaceutical preparations.

[0170] As used herein, unless otherwise stated, the terms “subject,” “individual,” and “patient” are used interchangeably and refer to humans as well as non-human mammals (e.g., non-human primates, canines, equines, felines, pigs, cattle, ungulates, lagomorphs, etc.). In various embodiments, the subject may be a person cared for by a physician or other healthcare worker in a hospital, outpatient clinic, or other clinical setting (e.g., adult male, adult female, adolescent male, adolescent female, boy, girl). In some embodiments, the subject may not be under the care or prescription of a physician or other healthcare worker.

[0171] As used herein, unless otherwise stated, the phrase “subject in need” means a subject who has or is at risk of a pathological condition requiring prevention or treatment with the compounds or salts described herein, as described below.

[0172] As used herein, unless otherwise stated, the terms “administered,” “applied,” “drugated,” and “administered” are defined as providing a subject with the composition via routes known in the art, including but not limited to intravenous, intra-arterial, oral, parenteral, oral, topical, transdermal, rectal, intramuscular, subcutaneous, intraosseous, mucosal, or intraperitoneal administration. In some embodiments, the composition may be administered orally. The terms “administered,” “applied,” “drugated,” and “administered” should be understood as providing an individual in need with a compound of the present invention or a prodrug of a compound of the present invention.

[0173] As used herein, and unless otherwise stated, the term "effective amount" or "therapeutic effective amount" refers to an amount of the compound described herein or a salt thereof sufficient to achieve the intended application, including but not limited to the treatment of a disease as defined below. Therapeutic effective amounts can vary depending on the intended application (in vitro or in vivo), the subject being treated, and the disease condition (e.g., the subject's weight and age, the severity of the disease condition, the route of administration, etc.), which can be readily determined by those skilled in the art. The term may also be applied to doses that can induce a specific response in target cells, such as reducing proliferation or downregulating the activity of target proteins. Specific doses can vary depending on the particular compound selected, the dosing regimen to be followed, whether it is administered in combination with other compounds, the timing of administration, the tissue of administration, and the physical delivery system on which it is delivered.

[0174] As used herein, unless otherwise stated, the terms “treatment” or “treated” refer to a method for obtaining a beneficial or anticipated outcome for a disease, condition, or medical condition, including but not limited to therapeutic and / or preventative benefits. In some embodiments, treatment or being treated involves administering a compound or composition disclosed herein to a subject. Therapeutic benefits may include eradication or improvement of the underlying disease being treated. Furthermore, therapeutic benefits may be obtained by eradicating or improving one or more physiological symptoms associated with the underlying disease, such as by observing improvement in the subject, even though the subject may still have the underlying disease. In some embodiments, to obtain preventative benefits, the composition is administered to a subject at risk of developing a particular disease, or to a subject who has reported one or more physiological symptoms of a disease, even if the disease may not yet be diagnosed. Treatment may include, for example, reducing, delaying, or alleviating the severity of one or more symptoms of a disease or condition, or it may include reducing the frequency with which a patient experiences symptoms such as a disease, defect, condition, or adverse condition. Herein, “treatment” may be used to refer to a method that results in some degree of treatment or improvement for a disease or condition, and may encompass a range of outcomes aimed at achieving that purpose, including but not limited to complete prevention of the condition.

[0175] As used herein, unless otherwise stated, the terms “treatment,” “cured,” and “therapy” mean the eradication or improvement of a disease or condition or one or more symptoms associated with a disease or condition. In some embodiments, these terms refer to the administration of one or more preventative or therapeutic agents to a subject suffering from such a disease or condition, thereby minimizing the spread or worsening of the disease or condition.

[0176] As used herein, unless otherwise stated, the term "prevention" means the use or administration of a compound provided herein, with or without other active compounds, prior to the onset of symptoms, particularly in patients at risk of developing the disease or condition described herein. The term "prevention" includes suppressing or alleviating symptoms of a specific disease. In some embodiments, patients with a family history of the disease are particularly candidates for prevention programs. Furthermore, patients with a history of recurrent symptoms are also potential candidates for prevention. In this regard, the term "prevention" may be used interchangeably with "preventive treatment."

[0177] It should be noted that if there is a difference between the described structure and its name, the described structure should be given greater weight. Furthermore, if the stereochemistry of a structure or part thereof is not indicated by, for example, thick or dashed lines, the structure or part thereof should be interpreted as encompassing all its stereoisomers.

[0178] Example

[0179] Some embodiments of the claimed subject matter are illustrated by the following non-limiting examples.

[0180] The disclosed compounds can generally be synthesized using the following general procedures or by appropriate combinations of commonly known synthetic methods. Based on this disclosure, the techniques for synthesizing these compounds are readily apparent and available to those skilled in the art. Many optionally substituted starting compounds and other reactants are commercially available or can be readily prepared by those skilled in the art using commonly used synthetic methods.

[0181] The following examples are intended to illustrate certain methods for preparing the disclosed compounds and are not intended to limit the range of reactions or reaction sequences that can be used to prepare the compounds provided herein.

[0182] The general procedure for preparing the inhibitor to be protected is shown below.

[0183] Option 1

[0184]

[0185] Option 2

[0186]

[0187] Example 1. Synthesis of N-((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0188]

[0189] Step 1: Add a suspension of (2S)-2-amino-2-(4-hydroxyphenyl)acetic acid methyl ester hydrochloride (19 g, 87.3 mmol) in dioxane (150 mL) to a solution of K₂CO₃ (24.13 g, 175 mmol) in water (300 mL). Then add (Boc)₂O (21.91 g, 100.3 mmol) at 0 °C and stir the reaction mixture at room temperature for 16 hours. Dilute with water (100 mL) and extract with ethyl acetate (200 mL x 3). The organic layers were combined, washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 3 / 2, v / v) to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid methyl ester (20 g, 71.1 mmol, yield: 81.4%) as a grayish-white solid. MS (ESI) m / z = 304.2 [M + Na] + .

[0190] Step 2: Platinum dioxide (0.82 g, 3.6 mmol) was added to a solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-hydroxyphenyl)acetic acid methyl ester (6.8 g, 24.2 mmol) in acetic acid (45 mL). The reaction mixture was stirred at 50 °C for 2 days under hydrogen at 0.3 MPa. The mixture was then filtered, and the filtrate was concentrated under vacuum to give a crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 2 / 1, v / v) to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-hydroxycyclohexyl)acetic acid methyl ester (4.4 g, 15.3 mmol, yield: 63.2%) as a pale yellow solid. MS (ESI) m / z = 310.3 [M + Na] + .

[0191] Step 3: At -78°C, add a solution of 5 mL of DCM containing 3.38 g (43.2 mmol) of dimethyl sulfoxide to a solution of 20 mL of DCM containing 2.74 g (21.6 mmol) of oxaloyl chloride, and stir the mixture at -78°C for 5–10 minutes. Then, slowly add a solution of 5 mL of DCM containing 3.1 g (10.8 mmol) of (S)-2-((tert-Butoxycarbonyl)amino)-2-(4-hydroxycyclohexyl)acetate, and stir the final mixture at -78°C for 1 hour. Quench the reaction with Et3N. Then dilute the mixture with water (50 mL) and adjust the pH to 3 with 1 N HCl. Extract with DCM (30 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 3 / 2, v / v) to give (S)-2-((tert-butyloxycarbonyl)amino)-2-(4-oxocyclohexyl)acetate (2.1 g, 7.4 mmol, yield: 68.5%) as a pale yellow oil. MS (ESI) m / z = 308.0 [M + Na] + .

[0192] Step 4: Under nitrogen atmosphere at -50°C, add a 4 mL solution of t-BuOK (1.49 g, 13.3 mmol) in DMF to a 16 mL solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-oxocyclohexyl)acetate (2.1 g, 7.4 mmol) and 2-(difluoromethane)sulfonylpyridine (1.72 g, 8.8 mmol). Then stir the mixture at room temperature for 2 hours. Dilute with water (200 mL) and extract with ethyl acetate (100 mL x 3). The organic layers were combined, washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 1, v / v) to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetate (1.5 g, 4.7 mmol, yield: 63.5%) as a pale yellow oil. MS (ESI) m / z = 342.1 [M+Na] + .

[0193] Step 5: LiOH·H₂O (79 mg, 1.88 mmol) was added to a THF / water solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetate (400 mg, 1.25 mmol) in a 1:1 (10 mL) THF / water mixture. The mixture was stirred overnight at room temperature. The mixture was then diluted with water (20 mL) and extracted with EA (20 mL). The aqueous solution was then adjusted to pH 3 with 1 N HCl. Extraction was performed with EA (20 mL * 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a colorless oily product (400 mg, crude). This was used directly for the next step. MS (ESI) m / z = 327.9 [M + Na] + .

[0194] Step 6: Stir a mixture of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetate (150 mg, 0.49 mmol), N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (172 mg, 0.49 mmol), HATU (280 mg, 0.74 mmol), and N,N-diisopropylethylamine (127 mg, 0.98 mmol) in THF (6 mL) overnight at room temperature. The product was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (elution gradient: DCM / MeOH = 8%, v / v) to give ((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobutyl-2-yl)phenyl)amino)-2-oxoethyl)carbamate tert-butyl ester (90 mg, 0.14 mmol, yield: 28.7%) as a yellow solid. MS (ESI) m / z = 638.3 [M+H] + .

[0195] Step 7: Add TFA (1 mL) to a 2 mL solution of ((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)carbamate (90 mg, 0.141 mmol) in DCM. Stir the mixture at room temperature for 2 hours. Then concentrate under vacuum to give crude N-((2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl)propionamide (110 mg, crude). No further purification is required for the next step. MS(ESI)m / z = 538.3 [M+H] + .

[0196] Step 8: The mixture of N-((2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl)propionamide (110 mg, 0.205 mmol), 2-ethylpyrazole-3-carboxylic acid (29 mg, 0.205 mmol), HATU (124 mg, 0.327 mmol), and N,N-diisopropylethylamine (106 mg, 0.818 mmol) in DMF (5 mL) was stirred at room temperature for 1 hour. It was diluted with water (50 mL) and extracted with ethyl acetate (30 mL x 3). The organic layers were combined, washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 1, v / v) to obtain Example 1 (13 mg, 0.020 mmol, yield: 9.6%) as a white solid. 1H NMR (400MHz, CD3OD) δ7.80(t,J=8.2Hz,1H),7.47(d,J=2.1Hz,1H),7.14–7.03(m,2H),6.84(d,J=2.1Hz,1H),4.91(d,J=10 .4Hz,1H),4.56(d,J=8.5Hz,1H),4.51(q,J=7.2Hz,2H),3.53-3.43(m,1H),3.40(t,J=4.9Hz,1H),3.22-3.09(m,2H),2.55- 2.45(m,2H),2.37–2.29(m,2H),2.28-2.18(m,2H),2.10(s,3H),2.06–1.93(m,2H),1.89–1.82(m,3H),1.65(dt,J=10.9,5 .5Hz,1H),1.35(t,J=7.2Hz,3H),1.31(d,J=7.0Hz,3H),1.29-1.14(m,4H),1.10(t,J=7.6Hz,3H).MS(ESI)m / z=660.3[M+H] + .

[0197] Example 2. Synthesis of N-((S)-2-((3-chloro-2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-1-cycloheptyl-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0198]

[0199] Step 1: Add m-CPBA (43.5 g, 180 mmol) to a toluene (100 mL) solution of 4-bromo-3-chloro-2-fluoroaniline (12 g, 53.8 mmol). Stir the mixture at 50 °C for 16 hours. After cooling, add DCM (100 mL) and filter the mixture. Wash the filtrate with 10% Na₂S₂SO₃ aqueous solution and purify by rapid chromatography (PE: EtOAc = 0–100%) to give 1-bromo-2-chloro-3-fluoro-4-nitrobenzene (5.3 g, 20.94 mmol, yield: 38.9%) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ7.88 (dd, J=9.0, 7.2Hz, 1H), 7.62 (dd, J=9.0, 2.0Hz, 1H).

[0200] Step 2: Under nitrogen atmosphere, a mixture of 1-bromo-2-chloro-3-fluoro-4-nitrobenzene (5.3 g, 20.8 mmol), tributyl(1-ethoxyvinyl)stanane (7.89 g, 21.8 mmol), and PdCl2(PPh3)2 (1.46 g, 2 mmol) in dioxane (100 mL) was stirred at 100 °C for 16 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0-2%) to give 2-chloro-1-(1-ethoxyvinyl)-3-fluoro-4-nitrobenzene (3.78 g, 15.4 mmol, yield: 74%) as a yellow oil. 1 H NMR(400MHz,)δ7.91(dd,J=8.7,6.9Hz,1H),7.36(dd,J=8.7,1.8Hz,1H),4.53(d ,J=3.0Hz,1H),4.46(d,J=3.0Hz,1H),3.96–3.88(m,2H),1.38(t,J=7.0Hz,3H).

[0201] Step 3: Concentrated hydrochloric acid (5 mL) was added to a THF (50 mL) solution of 2-chloro-1-(1-ethoxyvinyl)-3-fluoro-4-nitrobenzene (3.78 g, 15.4 mmol). The mixture was stirred at room temperature for 16 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0-8%) to give 1-(2-chloro-3-fluoro-4-nitrophenyl)ethyl ketone (2.1 g, 9.67 mmol, yield: 62.7%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ8.01 (dd, J=8.6, 6.5Hz, 1H), 7.43 (dd, J=8.6, 1.8Hz, 1H), 2.68 (s, 3H).

[0202] Step 4: At 0°C, NaBH4 (0.4 g, 10.6 mmol) was added to a methanol (50 mL) solution of 1-(2-chloro-3-fluoro-4-nitrophenyl)ethyl ketone (2.1 g, 9.7 mmol). The mixture was stirred at 0°C for 1 hour. The reaction was quenched with ice water and extracted with EA (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried with Na2SO4, and then concentrated under vacuum to give 1-(2-chloro-5-fluoro-4-nitrophenyl)ethanol (2 g, 9.1 mmol, yield: 93.8%) as a yellow oil. 1HNMR (400MHz, CDCl3) δ8.01(dd,J=8.7,7.0Hz,1H),7.63(dd,J=8.8,1.4Hz,1H),5.32(q,J=6.4Hz,1H),2.01(s,1H),1.51(d,J=6.4Hz,3H)

[0203] Step 5: At 0°C, PPh3 (2.86 g, 10.9 mmol) and CBr4 (3.62 g, 10.9 mmol) were added to a DCM (50 mL) solution of 1-(2-chloro-5-fluoro-4-nitrophenyl)ethanol (2 g, 9.1 mmol). The mixture was stirred at room temperature for 4 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0-10%) to give 1-(1-bromoethyl)-2-chloro-3-fluoro-4-nitrobenzene (2 g, 7.1 mmol, yield: 78%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ7.99 (dd, J=8.9, 7.1Hz, 1H), 7.59 (dd, J=8.9, 1.9Hz, 1H), 5.53 (q, J=6.9Hz, 1H), 2.04 (d, J=7.0Hz, 3H).

[0204] Step 6: Add NaOH (1.7 g, 44.2 mmol) to a DMF (30 mL) solution of 1-{[(2R)-1-benzylpyrrolidone-2-yl]carbonyl}-7-phenyl-5H-benzo[d]1-oxa-3,7-diaza-2-nickelhecyclononane-4-one (2.2 g, 4.4 mmol). Then, under N2, add a solution of 1-(1-bromoethyl)-2-chloro-3-fluoro-4-nitrobenzene (1.87 g, 6.62 mmol) dissolved in DMF (5 mL), and stir the mixture at room temperature for 20 minutes. The mixture was quenched with ice water (200 mL), then extracted with EA (80 mL x 3), and the combined organic layers were washed with brine (50 mL x 2). The mixture was dried over Na₂SO₄, concentrated, and purified by rapid chromatography (PE:EA = 0–100%) to give the product (1.4 g, 2.0 mmol, yield: 45%) as a red solid. MS (ESI) m / z = 699.0 [M+H] + .

[0205] Step 7: The mixture of 1-{[(2R)-1-benzylpyrrolidine-2-yl]carbonyl}-5-[(1S)-1-(2-chloro-3-fluoro-4-nitrophenyl)ethyl]-7-phenyl-5H-benzo[d]1-oxa-3,7-diaza-2-nickelcyclononane-4-one (1.4 g, 2 mmol) in 3NHCl / MeOH = 1:1 (14 mL) was stirred at 70 °C for 30 min. The mixture was then diluted with H2O (40 mL) and washed with DCM (10 mL x 3). Na2CO3 was added to the liquid layer to adjust the pH to 10, followed by the addition of THF (20 mL) and Boc2O (4.37 g, 20 mmol). The mixture was further stirred at room temperature for 16 h. The mixture was diluted with DCM (100 mL) and washed with water (50 mL x 3). The combined liquid layers were acidified to pH 3 with 1N HCl, washed with DCM (50 mL * 3), and the combined organic layers were concentrated to give (2R, 3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-3-fluoro-4-nitrophenyl)butyric acid (550 mg, 1.3 mmol, yield: 65%) as a colorless oil. MS (ESI) m / z = 399.0 [M + Na] + .

[0206] Step 8: Stir a mixture of (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-3-fluoro-4-nitrophenyl)butyric acid (550 mg, 1.46 mmol), 1-methylpiperazine (161 mg, 1.61 mmol), 1-hydroxybenzotriazole (256 mg, 1.90 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (364 mg, 1.90 mmol), and NMM (295 mg, 2.92 mmol) in THF (10 mL) at room temperature for 16 hours. The mixture was concentrated and purified by rapid chromatography (DCM:MA = 0–5%) to give N-[(2R,3S)-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate tert-butyl ester (580 mg, 1.2 mmol, yield: 82%) as a colorless oil. MS (ESI) m / z = 449.0 [M+H] + .

[0207] Step 9: The mixture of N-[(2R,3S)-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate tert-butyl ester (180 mg, 0.39 mmol) in 1,4-dioxane / HCl (4 M, 4 mL) and DCM (2 mL) was stirred at room temperature for 3 hours. The mixture was concentrated to give a white solid (2R,3S)-2-amino-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)but-1-one (190 mg, 0.37 mmol, crude). MS (ESI) m / z = 359.1 [M+H] + .

[0208] Step 10: Add propionyl chloride (49 mg, 0.53 mmol) to a DCM (6 mL) solution of (2R,3S)-2-amino-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)but-1-one (190 mg, 0.53 mmol) and N,N-diisopropylethylamine (274 mg, 2.12 mmol) at room temperature, and stir the mixture at room temperature for 2 hours. The mixture was diluted with DCM (40 mL), washed with water (20 mL) and brine (10 mL), dried over Na₂SO₄, concentrated, and purified by rapid chromatography (DCM:MA = 0–10%) to give N-[(2R,3S)-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (220 mg, 0.47 mmol, yield: 90%) as a yellow oil. MS (ESI) m / z = 415.1 [M+H] + .

[0209] Step 11: To a solution of N-[(2R,3S)-3-(2-chloro-3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (160 mg, 0.38 mmol) in ethanol (3 mL), add Raney nickel (0.5 mL), followed by hydrazine hydrate (0.5 mL). Stir the mixture at room temperature for 16 hours. Filter the mixture and concentrate the filtrate under vacuum to give N-[(2R,3S)-3-(4-amino-2-chloro-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (150 mg, 0.27 mmol, yield: 70%) as a brown oil. MS (ESI) m / z = 385.1 [M+H] + .

[0210] Step 12: At 0°C, add pyridine (53 mg, 0.67 mmol) and a solution of EA (215 mg, 0.67 mmol, 50%) of T3P to a solution of N-[(2R,3S)-3-(4-amino-2-chloro-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (130 mg, 0.34 mmol), (S)-cycloheptyl[(2-ethylpyrazol-3-yl)carbamoyl]acetic acid (92 mg, 0.34 mmol) in acetonitrile (6 mL). Stir the reaction mixture at 50°C for 16 hours. The mixture was diluted with EA (30 mL), washed with water (10 mL x 3) and brine (10 mL), dried over Na₂SO₄, concentrated, and purified by rapid chromatography (DCM:MA = 0–10%) to give N-[(S)-({3-chloro-2-fluoro-4-[(2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)(cycloheptyl)methyl] tert-butyl carbamate (60 mg, 0.056 mmol, yield: 16%), as a yellow oil. MS (ESI) m / z = 586.4 [M+H] + .

[0211] Step 13: The mixture of N-[(S)-({3-chloro-2-fluoro-4-[(2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)(cycloheptyl)methyl]tert-butyl carbamate (60 mg, 0.094 mmol) in TFA (1 mL) and DCM (1 mL) was stirred at room temperature for 1 hour. The mixture was concentrated to give a yellow oily crude product (55 mg, 0.092 mmol, crude product). MS (ESI) m / z = 538.3 [M+H] + .

[0212] Step 14: A mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptaylacetamyl]-2-chloro-3-fluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (55 mg, 0.10 mmol), 2-ethylpyrazole-3-carboxylic acid (29 mg, 0.20 mmol), HATU (78 mg, 0.20 mmol), and DIEA (106 mg, 0.82 mmol) in DMF (4 mL) was stirred at room temperature for 16 hours. The mixture was diluted with EA (40 mL), washed with water (15 mL x 3) and brine (15 mL), dried over Na2SO4, concentrated, and purified by preparative TLC (DCM: MeOH = 10%) to give Example 2 (9.1 mg, 0.013 mmol, yield: 13%) as a white solid.1 H NMR (400MHz, CD3OD) δ7.72(dd,J=8.5,7.6Hz,1H),7.39(d,J=2.1Hz,1H),7.13(dd,J=8.7,1.7Hz,1H),6.75(d ,J=2.1Hz,1H),5.12(d,J=9.3Hz,1H),4.53(d,J=8.2Hz,1H),4.46–4.40(m,2H),3.72–3.61(m,1H),3.58–3.5 0(m,2H),3.45–3.23(m,2H),2.45–2.30(m,2H),2.18–2.11(m,5H),2.10–2.02(m,3H),1.78–1.62(m,4H),1.5 6–1.36(m,8H),1.27(t,J=7.1Hz,3H),1.18(d,J=4.2Hz,3H),1.00(t,J=7.6Hz,3H).MS(ESI)m / z=660.8[M+H] + .

[0213] Example 3. Synthesis of N-((S)-2-((5-chloro-2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-1-cycloheptyl-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0214]

[0215] Step 1: Under nitrogen atmosphere, a mixture of 1-bromo-2-chloro-5-fluoro-4-nitrobenzene (8 g, 31.4 mmol), tributyl(1-ethoxyvinyl)stanane (11.91 g, 32.9 mmol), and PdCl2(PPh3)2 (2.2 g, 3.1 mmol) in dioxane (100 mL) was stirred at 100 °C for 16 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0-1%) to give 1-chloro-2-(1-ethoxyvinyl)-4-fluoro-5-nitrobenzene (4.8 g, 19.5 mmol, yield: 62.1%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ8.11(d,J=6.8Hz,1H),7.42(d,J=11.1Hz,1H),4.58-4.54(m,2H),3.93(q,J=7.0Hz,2H),1.40(t,J=7.0Hz,3H).

[0216] Step 2: HCl / 1,4-dioxane (4M, 25mL) was added to a solution of 1-chloro-2-(1-ethoxyvinyl)-4-fluoro-5-nitrobenzene (4.8 g, 19.5 mmol) in dioxane (25 mL). The mixture was stirred at room temperature for 16 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0–6.8%) to give 1-(2-chloro-5-fluoro-4-nitrophenyl)ethyl ketone (3.5 g, 16.1 mmol, yield: 82.5%) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ8.15 (d, J = 6.3Hz, 1H), 7.46 (d, J = 10.2Hz, 1H), 2.68 (s, 3H).

[0217] Step 3: Add NaBH4 (670 mg, 17.7 mmol) to a solution of 1-(2-chloro-5-fluoro-4-nitrophenyl)ethyl ketone (3.5 g, 16.1 mmol) in MeOH, and stir the mixture at 0 °C for 2 hours. Quench the mixture with ice water and extract with EtOAc (50 mL x 3). Wash the combined organic layers with brine, dry with Na2SO4, and concentrate under vacuum to give 1-(2-chloro-5-fluoro-4-nitrophenyl)ethanol (3.4 g, 15.5 mmol, yield: 96.2%) as a brown oil. 1 H NMR (400MHz, CDCl3) δ8.07(d,J=6.6Hz,1H),7.64(d,J=11.6Hz,1H),5.36 -5.20(m,1H),2.17(d,J=3.6Hz,1H),1.50(d,J=6.4Hz,3H).

[0218] Step 4: At 0°C, PPh3 (4.88 g, 18.6 mmol) and CBr4 (6.17 g, 18.6 mmol) were added to a DCM (50 mL) solution of 1-(2-chloro-5-fluoro-4-nitrophenyl)ethanol (3.4 g, 15.5 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (PE / EtOAc = 0-2%) to give 1-(1-bromoethyl)-2-chloro-5-fluoro-4-nitrobenzene (3.3 g, 11.7 mmol, yield: 75.4%) as a yellow oil. 1 H NMR (400MHz, CDCl3) δ8.12(d,J=6.8Hz,1H),7.59(d,J=11.4Hz,1H),5.49(qd,J=6.9,1.2Hz,1H),2.03(d,J=6.9Hz,3H).

[0219] Step 5: Add NaOH (1.6 g, 40 mmol) to a DMF (30 mL) solution of 1-{[(2R)-1-benzylpyrrolidine-2-yl]carbonyl}-7-phenyl-5H-benzo[d]1-oxa-3,7-diaza-2-nickelcyclononane-4-one (2 g, 4 mmol). Then, under N2, add a DMF (5 mL) solution of 1-(1-bromoethyl)-2-chloro-5-fluoro-4-nitrobenzene (3.39 g, 12 mmol) and stir the mixture at 0 °C for 20 minutes. The mixture was quenched with ice water (100 mL), then extracted with EA (50 mL x 3). The combined organic layers were washed with brine (50 mL x 2), dried over Na2SO4, concentrated, and rapidly purified by chromatography (PE:EA = 0–100%) to give 1-{[(2R)-1-benzylpyrrolidine-2-yl]carbonyl}-5-[(1S)-1-(2-chloro-5-fluoro-4-nitrophenyl)ethyl]-7-phenyl-5H-benzo[d]1-oxa-3,7-diaza-2-nickel-cyclononane-4-one (0.69 g, 0.98 mmol, yield: 24.5%) as a red solid. MS (ESI) m / z = 699.0, 701.0 [M+H] + .

[0220] Step 6: Add 10 mL of 3N HCl to a MeOH solution of 1-{[(2R)-1-benzylpyrrolidine-2-yl]carbonyl}-5-[(1S)-1-(2-chloro-5-fluoro-4-nitrophenyl)ethyl]-7-phenyl-5H-benzo[d]1-oxa-3,7-diaza-2-nickelhecyclononane-4-one (600 mg, 0.85 mmol) and stir the mixture at 70 °C for 1 hour. After the red color of the Ni(II) complex disappears, evaporate the mixture to dryness under vacuum. Add 10 mL of H2O, treat the resulting mixture with excess NH4OH and extract with DCM. Evaporate the aqueous phase under vacuum and purify by preparative HPLC (20-70, 0.1% FA) to obtain the desired product (110 mg, 0.40 mmol, yield: 46.3%) as a grayish-white solid. MS(ESI)m / z = 277.1, 279.1 [M+H] + .

[0221] Step 7: Add NaHCO3 (334 mg, 3.9 mmol) and (Boc)2O (434 mg, 2 mmol) to a 1:1 (10 mL) solution of (2R,3S)-2-amino-3-(2-chloro-5-fluoro-4-nitrophenyl)butyric acid (110 mg, 0.39 mmol) in dioxane / water. Stir the mixture at room temperature for 16 hours. Dilute the mixture with EtOAc and wash with water (20 mL x 3). Acidify the combined aqueous layers to pH 6 with 1N HCl and extract with DCM (20 mL x 3). Concentrate the combined organic layers to give 2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-5-fluoro-4-nitrophenyl)butyric acid (150 mg, 0.39 mmol, yield: 100%) as a yellow oil. MS(ESI)m / z = 399.1 [M + Na] + .

[0222] Step 8: Stir the mixture of (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-5-fluoro-4-nitrophenyl)butyric acid (160 mg, 0.42 mmol), 1-methylpiperazine (42.54 mg, 0.42 mmol), HOBt (114.77 mg, 0.84 mmol), EDCI (162.83 mg, 0.84 mmol), and NMM (128.88 mg, 1.27 mmol) in THF (10 mL) at room temperature for 3 hours. The mixture was concentrated and rapidly purified by chromatography (DCM:MeOH = 0–5%) to give N-[(2R,2S)-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate (150 mg, 0.33 mmol, yield: 76.9%) as a yellow oil. MS (ESI) m / z = 459.1 [M+H] + .

[0223] Step 9: Add dioxane (4 M, 5 mL) of HCl to a DCM (5 mL) solution of N-[(2R,3S)-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate (150 mg, 0.32 mmol). Stir the mixture at room temperature for 4 hours. Concentrate the mixture under vacuum to give a white solid 2R,3S)-2-amino-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)but-1-one (130 mg, crude). MS (ESI) m / z = 359.0 [M+H] + .

[0224] Step 10: Add DIPEA (234 mg, 1.8 mmol) and propionyl chloride (30.5 mg, 0.33 mmol) to a DCM (10 mL) solution of (2R, 3S)-2-amino-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)but-1-one (130 mg, 0.33 mmol). Stir the reaction mixture at room temperature for 0.5 hours. Dilute the mixture with water (20 mL) and extract with DCM (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (CH₂Cl₂ / MeOH = 0–7%) to give N-[(2R,3S)-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (110 mg, 0.27 mmol, yield: 73.2%) as a white solid. MS (ESI) m / z = 415.1 [M+H] + .

[0225] Step 11: To a 5 mL ethanol (5 mL) solution of N-[(2R,3S)-3-(2-chloro-5-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (100 mg, 0.24 mmol), Raney nickel (1.5 mL) was added, followed by hydrazine hydrate (1.5 mL). The mixture was stirred at room temperature for 5 hours. The mixture was filtered, and the filtrate was concentrated under vacuum to give N-[(2R,3S)-3-(4-amino-2-chloro-5-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (90 mg, 0.23 mmol, yield: 97%) as a yellow oil. MS (ESI) m / z = 385.1 [M+H] + .

[0226] Step 12: At 0°C, add pyridine (31 mg, 0.39 mmol) and a solution of EA (124 mg, 0.39 mol, 50%) of T3P to a solution of N-[(2R,3S)-3-(4-amino-2-chloro-5-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (75 mg, 0.19 mmol), (S)-cycloheptayl[(2-ethylpyrazol-3-yl)carboxamide]acetic acid (106 mg, 0.39 mmol) in acetonitrile (4 mL). Stir the reaction mixture at 50°C for 16 hours. Dilute the mixture with water (20 mL) and extract with EA (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by preparative TLC (DCM / MeOH = 0-10%) to give N-[(S)-({5-chloro-2-fluoro-4-[(2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)(cycloheptyl)methyl] tert-butyl carbamate (9 mg, 0.01 mmol, yield: 5%), as a yellow oil. MS (ESI) m / z = 638.3 [M+H] + .

[0227] Step 13: N-[(S)-({5-chloro-2-fluoro-4-[(2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)(cycloheptyl)methyl]tert-butyl carbamate (15 mg, 0.0235 mmol) was stirred in TFA (1 mL) and DCM (1 mL) at room temperature for 1 h. The mixture was concentrated to give a yellow oily N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptylacetamido]-2-chloro-5-fluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (20 mg, crude). MS (ESI) m / z = 538.2 [M+H] + .

[0228] Step 14: Stir the mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptylacetamido]-2-chloro-5-fluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (20 mg, 0.037 mmol), 2-ethylpyrazole-3-carboxylic acid (10 mg, 0.074 mmol), HATU (28 mg, 0.074 mol), and DIEA (38 mg, 0.30 mmol) in DMF (2 mL) at room temperature for 16 hours. The mixture was diluted with EA (40 mL), washed with water (15 mL * 3) and brine (15 mL), dried with Na2SO4, concentrated, and purified by preparative HPLC (Gemini-C18 150 x 21.2 mm, 5 μm: ACN-H2O (0.1% FA) 15-45) to obtain Example 3 (2.9 mg, 0.0038 mmol, yield: 10.4% diformate), as a white solid. 1 H NMR (400MHz, CD3OD) δ8.39(s,2H),7.95(d,J=7.2Hz,1H),7.40(d,J=2.1Hz,1H),7.21(d,J=11.8Hz,1H),6 .76(d,J=2.1Hz,1H),5.11(d,J=8.7Hz,1H),4.52(d,J=8.3Hz,1H),4.48–4.40(m,2H),3.67-3.61(m,1H), 3.59-3.53(m,2H),3.44-3.31(m,2H),2.45–2.31(m,2H),2.17(s,3H),2.15–2.07(m,5H),1.77–1.62(m,4 H),1.54–1.39(m,8H),1.29(d,J=7.2Hz,3H),1.20(s,3H),1.00(t,J=7.6Hz,3H).MS(ESI)m / z=660.2[M+H] + .

[0229] Example 4. Synthesis of N-((S)-1-cycloheptyl-2-((2,3-difluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0230]

[0231] Step 1: A mixture of 1-bromo-2,3-difluoro-4-nitrobenzene (4.5 g, 18.9 mmol), tributyl(1-ethoxyvinyl)stanane (8.19 g, 22.6 mmol), and bis(triphenylphosphine)palladium(II) chloride (0.93 g, 1.3 mmol) in dioxane (100 mL) was stirred overnight at 100 °C under argon atmosphere. The mixture was then concentrated under vacuum to give a crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate = 10:1, v / v) to give compound 1-(1-ethoxyvinyl)-2,3-difluoro-4-nitrobenzene (3.6 g, 71.1 mmol, yield: 81.4%) as a yellow solid. MS (ESI) m / z = 230.0 [M+H] + .

[0232] Step 2: Add 4M HCl / dioxane (20 mL) to a solution of 1-(1-ethoxyvinyl)-2,3-difluoro-4-nitrobenzene (3.6 g, 15.7 mmol) in dioxane (20 mL). Stir the mixture overnight at room temperature. Then concentrate under vacuum to give a crude product, which is purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate = 10:1, v / v) to give compound 1-(2,3-difluoro-4-nitrophenyl)ethyl-1-one (2.3 g, 11.4 mmol, yield: 72.6%) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ7.94–7.86(m,1H),7.77(ddd,J=8.8,6.3,2.2Hz,1H),2.72(d,J=4.7Hz,3H).

[0233] Step 3: At 0°C, sodium borohydride (0.47 g, 12.5 mmol) was added to a methanol (30 mL) solution of 1-(2,3-difluoro-4-nitrophenyl)ethyl ketone (2.3 g, 11.4 mmol). The mixture was stirred at room temperature for 1.5 hours. The mixture was quenched with ice water (100 mL) and extracted with ethyl acetate (70 mL x 3). The organic layers were combined, washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give compound 1-(2,3-difluoro-4-nitrophenyl)ethyl-1-ol (2.2 g, 10.8 mmol, yield: 94.7%) as a brown oil. 1 H NMR (400MHz, CDCl3) δ7.89 (ddd, J=8.7, 6.5, 2.1Hz, 1H), 7.59–7.42 (m, 1H), 5.28 (q, J=6.5Hz, 1H), 1.55 (d, J=6.5Hz, 3H).

[0234] Step 4: At 0 °C, triphenylphosphine (3.4 g, 12.9 mmol) and tetrabromomethane (4.3 g, 12.9 mmol) were added to a solution of 1-(2,3-difluoro-4-nitrophenyl)ethanol (2.2 g, 10.8 mmol) in DCM (40 mL). The mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum to give a crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate = 20:1, v / v) to give compound 1-(1-bromoethyl)-2,3-difluoro-4-nitrobenzene (2.4 g, 9.0 mmol, yield: 83.3%) as a pale yellow oil. 1 H NMR (400MHz, CDCl3) δ7.89 (ddd, J=8.8, 6.6, 2.1Hz, 1H), 7.46 (ddd, J=8.8, 6.4, 2.1Hz, 1H), 5.40 (q, J=7.0Hz, 1H), 2.06 (d, J=7.0Hz, 3H).

[0235] Step 5: With stirring, 0.8 g (20 mmol) of powdered sodium hydroxide was added to a 24 mL solution of (SP-4-4)-[N-[phenyl[2-[[[(1R,2S)-1-(phenylmethyl)-2-pyrrolyl-κN]carbonyl]amino-κN]-phenyl]methylene]glycine(2-)-κN,κO]nickel (2.5 g, 5 mmol) in DMF. Then, a 6 mL solution of 1-(1-bromoethyl)-2,3-difluoro-4-nitrobenzene (2.2 g, 8.5 mmol) in DMF was added to the solution. The mixture was stirred under nitrogen, and the reaction progress was monitored by TLC. The mixture was diluted with 300 mL of 0.3 N HCl and extracted with EA (100 mL x 3). The combined organic layers were washed with brine (150 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate = 1:2, v / v) to give compound 8 (1.2 g, 1.8 mmol, yield: 36.0%) as a deep red solid. MS (ESI) m / z = 683.1 [M+H] + .

[0236] Steps 6 and 7: Add 3M HCl / MeOH = 1:1 (15 mL) to a MeOH solution of compound 8 (1200 mg, 1.76 mmol) and stir the mixture at 70 °C for 15 min. Dilute the mixture with water (50 mL) and wash with DCM (30 mL x 3). Add Na₂CO₃ to the aqueous phase to adjust pH to 10, and add THF (40 mL) and Boc₂O (3833 mg, 17.56 mmol). Stir the mixture further at room temperature for 16 h. Dilute the mixture with EA (80 mL) and wash with water (50 mL x 3). Acidify the combined liquid layers to pH 3 with 1N HCl, wash with EA (20 mL x 3), and concentrate the combined organic layers to give the crude product (400 mg, 1.11 mmol, yield: 63.2%) as a colorless oil. No further purification is required for the next step. MS(ESI)m / z = 383.0 [M+Na] + .

[0237] Step 8: A mixture of (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2,3-difluoro-4-nitrophenyl)butyric acid (270 mg, 0.75 mmol), 1-methylpiperazine (83 mg, 0.82 mmol), HOBT (132 mg, 0.97 mmol), EDCI (187 mg, 0.97 mmol), and NMM (152 mg, 1.50 mmol) in THF (8 mL) was stirred at room temperature for 16 hours. The mixture was concentrated and purified by rapid chromatography (DCM:MeOH = 0–5%) to give the product (250 mg, 0.53 mmol, yield: 71.6%) as a yellow oil. MS (ESI) m / z = 443.2 [M+H] + .

[0238] Step 9: A mixture of N-[(2R,3S)-3-(2,3-difluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate tert-butyl ester (250 mg, 0.565 mmol) in DCM (5 mL) and HCl in dioxane (4 M, 5 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated to give a crude product (240 mg, crude product) as a white solid. MS (ESI) m / z = 342.9 [M+H] + .

[0239] Step 10: At room temperature, propionyl chloride (65 mg, 0.70 mmol) was added to a DCM (5 mL) solution of (2R,3S)-2-amino-3-(2,3-difluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)but-1-one (240 mg, 0.70 mmol) and DIEA (362 mg, 2.80 mmol), and the mixture was stirred at 25 °C for 2 hours. The mixture was diluted with DCM (40 mL), washed with water (10 mL) and brine (10 mL), dried over Na2SO4, concentrated, and purified by rapid chromatography (DCM:MeOH = 0–10%) to give the product (150 mg, 0.36 mmol, yield: 51.0%) as a white solid. MS (ESI) m / z = 399.1 [M+H] + .

[0240] Step 11: A mixture of N-[(2R,3S)-3-(2,3-difluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (150 mg, 0.38 mmol) and Pd / C (30 mg) in THF (10 mL) was stirred at 25 °C for 16 hours. The mixture was filtered through diatomaceous earth, concentrated, and then purified by rapid chromatography (DCM:MeOH = 0-5%) to give the product (100 mg, 0.26 mmol, yield: 68.5%) as a yellow oil. MS (ESI) m / z = 369.2 [M+H] + .

[0241] Step 12: At 0 °C, a solution of pyridine (32 mg, 0.4072 mmol) and T3P in EA (259 mg, 0.40 mmol, 50% wt) was added to a solution of N-[(2R,3S)-3-(4-amino-2,3-difluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]acrylamide (75 mg, 0.20 mmol), (S)-{[(tert-butoxy)carbonyl]amino}(cycloheptyl)acetic acid (110 mg, 0.40 mmol) in ACN (4 mL). The reaction mixture was stirred at 50 °C for 16 hours. The mixture was diluted with water (50 mL) and extracted with EA (20 mL * 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, concentrated under vacuum, and purified by preparative TLC (DCM / MeOH = 10%) to obtain the product. (12 mg, 0.017 mmol, yield: 8.5%), as a yellow solid. MS (ESI) m / z = 622.3 [M+H] + .

[0242] Step 13: The mixture of N-[(S)-cycloheptyl({2,3-difluoro-4-[(2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)methyl]tert-butyl carbamate (12 mg, 0.019 mmol) in TFA (0.2 mL) / DCM (0.1 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily product (15 mg, crude product). MS (ESI) m / z = 522.3 [M+H] + .

[0243] Step 14: A mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptylacetamido]-2,3-difluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (15 mg, 0.0288 mmol), 2-ethylpyrazole-3-carboxylic acid (4 mg, 0.0288 mmol), HATU (22 mg, 0.0576 mmol), and DIEA (30 mg, 0.23 mmol) in THF (3 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by preparative TLC (DCM:MeOH = 10%) to give Example 4 (5.2 mg, 0.0073 mmol, yield: 25.3%) as a white solid. 1 H NMR(400MHz,CD3OD)δ–7.50-7.45(m,1H),7.39(d,J=1.8Hz,1H),7.04-7.00(m,1H),6.7 5(d,J=1.7Hz,1H),5.04(d,J=9.7Hz,1H),4.51(d,J=8.1Hz,1H),4.43(q,J=7.2Hz,2H), 3.56–3.28(m,5H),2.56–2.36(m,2H),2.23(s,3H),2.19–2.05(m,5H),1.77–1.60(m,4H ),1.55–1.37(m,8H),1.27-1.22(m,6H),1.02(t,J=7.6Hz,3H).MS(ESI)m / z=644.2[M+H] + .

[0244] Examples 5 and 6. Synthesis of N-((S)-1-cycloheptyl-2-((2-fluoro-4-((2S,3R)-4-((1S,6R)-5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide and N-((S)-1-cycloheptyl-2-((2-fluoro-4-((2S,3R)-4-((1R,6S)-5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0245]

[0246] Step 1: At 0 °C, AcOH (27 mg, 0.45 mmol) was added to a MeOH (15 mL) solution of 2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tert-butyl ester (900 mg, 4.54 mmol) and HCHO (735 mg, 9.08 mmol, 37% content), and the mixture was stirred at 0 °C for 20 minutes. Then, sodium cyanoborohydride (571 mg, 9.08 mmol) was added at 0 °C, and the final mixture was stirred overnight at room temperature. The reaction mixture was quenched with ice water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to give compound 5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tert-butyl ester (800 mg, 3.77 mmol, yield: 83.0%) as a colorless oil. 1 H NMR (400MHz, CDCl3) δ3.80–3.57(m,1H),3.07–2.83(m,2H),2.71–2.60(m,1H),2.57–2.46(m, 1H),2.43(s,3H),2.21-2.12(m,1H),1.49(s,9H),0.66–0.48(m,2H).MS(ESI)m / z=213.2[M+H] + .

[0247] Step 2: Add a solution of dioxane (6 mL) in 4N HCl to a DCM (10 mL) solution of 800 mg (3.77 mmol) of 5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-carboxylic acid tert-butyl ester (DCM). Stir the mixture overnight at room temperature. Then concentrate the mixture under vacuum to give compound 2-methyl-2,5-diazabicyclo[4.1.0]heptane (750 mg, crude product) as a white solid. MS (ESI) m / z = 113.1 [M+H] + .

[0248] Step 3: 2-Methyl-2,5-diazabicyclo[4.1.0]heptane (357 mg, 1.93 mmol), (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (330 mg, 0.96 mmol), 1-hydroxybenzotriazole (195 mg, 1.45 mmol), N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (277 mg, 1.45 mmol), and NMM (488 mg, The mixture of 4.82 mmol of carbamate in 12 mL of THF was stirred at room temperature for 3 hours, then concentrated under vacuum to give a crude product, which was purified by rapid chromatography (elution gradient: dichloromethane / methanol, 25 / 1, v / v) to give compound N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate (350 mg, 0.80 mmol, yield: 83.2%) as a pale yellow solid. MS (ESI) m / z = 437.1 [M+H] + .

[0249] Step 4: Add 4N HCl / dioxane (5 mL) to a DCM solution of N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]carbamate tert-butyl ester (350 mg, 0.80 mmol) (5 mL). Stir the mixture overnight at room temperature. Then concentrate the mixture under vacuum to give a pale yellow solid compound (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}but-1-one (350 mg, crude). No further purification is required for the next step. MS (ESI) m / z = 337.1 [M+H] + .

[0250] Step 5: Add propionyl chloride (116 mg, 1.25 mmol) to a DCM (10 mL) solution of (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}but-1-one (350 mg, 1.04 mmol) and N,N-diisopropylethylamine (672 mg, 5.20 mmol), and stir the mixture at room temperature for 2 minutes. The solution was then concentrated under vacuum for hours to obtain a crude product, which was purified by rapid chromatography (elution gradient: dichloromethane / methanol, 25 / 1, v / v) to give N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}-1-oxobut-2-yl]propionamide (350 mg, 0.89 mmol, yield: 85.7%) as a yellow solid. MS (ESI) m / z = 393.1 [M+H] + .

[0251] Step 6: Palladium / carbon (70 mg) was added to a 1 / 1 (10 mL) solution of N-[(2R,3S)-3-(3-fluoro-4-nitrophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}-1-oxobut-2-yl]propionamide (350 mg, 0.89 mmol) in EA / THF = 1 / 1, and the mixture was stirred overnight at 30 °C. The mixture was then filtered through diatomaceous earth. The filtrate was concentrated under vacuum to give a light brown oily product, N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}-1-oxobut-2-yl]propionamide (360 mg, crude). MS (ESI) m / z = 363.1 [M+H] + .

[0252] Step 7: The mixture of N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-{5-methyl-2,5-diazabicyclo[4.1.0]hept-2-yl}-1-oxobut-2-yl]propionamide (360 mg, 0.99 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptaacetic acid (539 mg, 1.99 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)ureoyl (755 mg, 1.95 mmol), and N,N-diisopropylethylamine (1027 mmol, 7.95 mmol) in DMF (15 mL) was stirred overnight at 30 °C. It was diluted with water (150 mL) and extracted with ethyl acetate (80 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: dichloromethane / methanol, 15 / 1, v / v) to give N-[(S)-cycloheptyl({2-fluoro-4-[(2S,3R)-4-{5-methyl-2,5-diazabicyclo[4.1.0]heptan-2-yl}-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)methyl] tert-butyl carbamate (210 mg, 0.34 mmol, yield: 34.3%), as a brown solid. MS (ESI) m / z = 616.3 [M+H] + .

[0253] Step 8: Add TFA (2 mL) to a DCM (3 mL) solution of N-[(S)-cycloheptyl({2-fluoro-4-[(2S,3R)-4-{5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl}-4-oxo-3-propamidobut-2-yl]phenyl}carbamoyl)methyl]carbamate (210 mg, 0.34 mmol) and stir the mixture at room temperature for 1.5 hours. Then concentrate under vacuum to give N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptylacetamido]-3-fluorophenyl}-1-{5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl}-1-oxobut-2-yl]propionamide (240 mg, crude), a brown solid. No further purification is required for the next step. MS(ESI)m / z = 516.3[M+H] + .

[0254] Step 9: The mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptaylacetamyl]-3-fluorophenyl}-1-{5-methyl-2,5-diazabicyclo[4.1.0]heptane-2-yl}-1-oxobut-2-yl]propionamide (110 mg, 0.21 mmol), 2-ethylpyrazole-3-carboxylic acid (39 mg, 0.28 mmol), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)ureoyl (162 mg, 0.43 mmol), and N,N-diisopropylethylamine (165 mg, 1.28 mmol) in DMF (6 mL) was stirred at room temperature for 2 hours. It was diluted with water (60 mL) and extracted with ethyl acetate (40 mL x 3). The organic layers were combined, dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a crude product. The crude product was purified by rapid chromatography (elution gradient: dichloromethane / methanol, 15 / 1, v / v), preparative HPLC (mobile phase: ACN-H2O (0.1% FA), 35 / 65-60 / 40), and chiral HPLC to obtain the product. In Examples 5 (24.9 mg, 0.039 mmol, yield: 18.3%) and 6 (46.7 mg, 0.073 mmol, yield: 34.3%), the product was a white solid.

[0255] Example 5 11H NMR (400 MHz, CD3OD) δ 7.80–7.72 (m, 1H), 7.46 (d, J = 2.1 Hz, 1H), 7.16–7.04 (m, 2H), 6.82 (t, J = 2.0 Hz, 1H), 5.30 (d, J = 9.5 Hz, 0.6H), 4.88 (d, J = 10.8 Hz, 0.6H), 4.59 (dd, J = 8.2, 3.4 Hz, 1H), 4.51 (q, J = 7.2 Hz, 2H), 3.96–3.85 (m, 0.6H), 3.59–3.47 (m, 0.3H), 3.24–2.97 (m, 1.3H), 2.81 (td, J = 7.5, 4.1 Hz, 0.4H), 2.57–2.33 (m, 5H), 2.32–2.18 (m, 3H), 2.17–2.06 (m, 3H), 1.87–1.78 (m, 2H), 1.77–1.68 (m, 2H), 1.67–1.39 (m, 10H), 1.37–1.31 (m, 6H), 1.29–1.14 (m, 1H), 1.14–1.07 (m, 3H), 0.79 (q, J = 6.7 Hz, 0.6H), 0.75–0.69 (m, 0.6H), 0.47 (q, J = 7.1 Hz, 0.4H), -0.02–-0.08 (dd, J = 10.2, 6.0 Hz, 0.3H). MS (ESI) m / z = 638.3 [M+H] + .

[0256] Example 6 1 1H NMR (400 MHz, CD3OD) δ 7.71 (t, J = 8.3 Hz, 1H), 7.46 (d, J = 2.1 Hz, 1H), 7.12–7.03 (m, 2H), 6.82 (d, J = 2.1 Hz, 1H), 5.07 (d, J = 10.5 Hz, 1H), 4.62–4.57 (m, 1H), 4.50 (q, J = 7.2 Hz, 2H), 3.75 (dt, J = 13.2, 3.0 Hz, 1H), 3.28–3.22 (m, 2H), 2.69–2.60 (m, 1H), 2.52–2.41 (m, 2H), 2.31–2.24 (m, 2H), 2.21 (s, 3H), 2.18–2.11 (m, 1H), 1.87–1.77 (m, 2H), 1.77–1.69 (m, 2H), 1.66–1.54 (m, 4H), 1.53–1.39 (m, 5H), 1.38–1.32 (m, 6H), 1.14–1.09 (m, 3H), 0.48 (q, = 6.8 Hz, 1H), -0.29–-0.39 (m, 1H). MS (ESI) m / z = 638.3 [M+H] +.

[0257] Example 7. Synthesis of N-((S)-1-cycloheptyl-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidinyl-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0258]

[0259] Step 1: At -40°C under nitrogen, a solution of t-BuOK (1.8 g, 16 mmol) in DMSO (10 mL) was added to a DMF (20 mL) solution of 4-oxopiperidinium-1-carboxylic acid tert-butyl ester (2 g, 10 mmol) and DFMPS (1.55 g, 8 mmol). The mixture was stirred at 25°C for 2 hours. The mixture was neutralized with 1N HCl, diluted with EA (120 mL), washed with water (60 mL x 2) and brine (60 mL x 2), dried over Na2SO4, concentrated, and purified by rapid chromatography (PE:EA = 0–3%) to give the product (1.7 g, 6.9 mmol, yield: 69.0%) as a white solid. 1 H NMR (400MHz, CDCl3) δ3.45–3.36(m,4H),2.16-2.13(m,4H),1.47(s,9H).

[0260] Step 2: The mixture of 4-(difluoromethylene)piperidine-1-carboxylic acid tert-butyl ester (500 mg, 2.14 mmol) in HCl-dioxane (4 M, 4 mL) / DCM (4 mL) was stirred at 25 °C for 2 hours. The mixture was concentrated to give a white solid crude product (280 mg, crude product). MS (ESI) m / z = 134.1 [M+H] + .

[0261] MS(ESI)m / z = 134.1 [M+H] + .

[0262] Step 3: A mixture of (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(3-fluoro-4-nitrophenyl)butyric acid (600 mg, 1.75 mmol), 4-(difluoromethylene)piperidine (297 mg, 1.75 mmol), HOBt (308 mg, 2.28 mmol), EDCI (437 mg, 2.28 mmol), and NMM (355 mg, 3.505 mmol) in THF (8 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and rapidly purified by chromatography (PE:EA = 0–50%) to give the product (620 mg, 1.28 mmol, yield: 73.4%) as a pale yellow solid. MS (ESI) m / z = 480.1 [M+H] + .

[0263] Step 4: The mixture of N-[(2R,3S)-1-[4-(difluoromethylene)piperidin-1-yl]-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl]carbamate (620 mg, 1.355 mmol) in HCl dioxane (6 mL) / DCM (2 mL) was stirred at 25 °C for 3 hours. The mixture was concentrated to give a white solid crude product (640 mg, crude product). MS (ESI) m / z = 358.0 [M+H] + .

[0264] Step 5: At room temperature, propionyl chloride (166 mg, 1.79 mmol) was added to a solution of (2R,3S)-2-amino-1-[4-(difluoromethylene)piperidin-1-yl]-3-(3-fluoro-4-nitrophenyl)but-1-one (640 mg, 1.79 mmol) and DIEA (926 mg, 7.16 mmol) in DCM (8 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was diluted with DCM (40 mL), washed with water (20 mL) and brine (10 mL), dried over Na2SO4, concentrated, and purified by rapid chromatography (PE:EA = 0–60%) to give the product (510 mg, 1.05 mmol, yield: 58.5%) as a yellow oil. MS (ESI) m / z = 414.0 [M+H] + .

[0265] Step 6: Add Raney nickel (0.5 mL) to a 3 mL ethanol (3 mL) solution of N-[(2R,3S)-1-[4-(difluoromethylene)piperidin-1-yl]-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl]propionamide (80 mg, 0.19 mmol), followed by hydrazine hydrate (0.5 mL). Stir the mixture at room temperature for 5 hours. Filter the mixture and concentrate the filtrate under vacuum to give the crude product (50 mg, 0.12 mmol, yield: 60.7%) as a yellow oil. MS (ESI) m / z = 384.2 [M+H] + .

[0266] Step 7: A mixture of N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (50 mg, 0.13 mmol), (S)-{[(tert-butoxy)carbonyl]amino}(cycloheptyl)acetic acid (35 mg, 0.13 mmol), HATU (99 mg, 0.26 mmol), and DIEA (50 mg, 0.39 mmol) in DMF (3 mL) was stirred at room temperature for 16 hours. The mixture was concentrated and purified by rapid chromatography (PE:EA = 0–60%) to give the product (80 mg, 0.088 mmol, yield: 67.4%) as a yellow solid. MS (ESI) m / z = 637.2 [M+H] + .

[0267] Step 8: The mixture of N-[(S)-cycloheptyl({4-[(2S,3R)-4-[4-(difluoromethylene)piperidin-1-yl]-4-oxo-3-propamidobutyl-2-yl]-2-fluorophenyl}carbamoyl)methyl] tert-butyl carbamate (80 mg, 0.1256 mmol) in TFA (1 mL) / DCM (0.5 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily crude product (90 mg, crude product). MS (ESI) m / z = 537.2 [M+H] + .

[0268] Step 9: Stir the mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-cycloheptylacetamido]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (90 mg, 0.17 mmol), 2-ethylpyrazole-3-carboxylic acid (24 mg, 0.17 mol), HATU (127 mg, 0.335 mmol), and DIEA (173 mg, 1.34 mmol) in DMF (4 mL) at room temperature for 16 hours. The mixture was diluted with EA (40 mL) and washed with water (15 mL * 3) and brine (15 mL). It was dried with Na2SO4, concentrated, and purified by preparative TLC (DCM:MeOH = 20:1) to obtain the product (25 mg). The product was then purified by chiral HPLC (:-SP-120-10-C18-BIO-C18 250 x 50 mm, 10 μm (pH 8-10): CO2 MeOH (DEA)) to obtain Example 7 (12 mg, 0.0182 mmol, yield: 10.8%) as a white solid.

[0269] 1 H NMR (400MHz, CD3OD) δ7.81-7.76(m,1H),7.51(d,J=2.1Hz,1H),7.18–7.11(m,2H),6.87(d,J=2. 1Hz,1H),4.99(d,J=10.3Hz,1H),4.63(d,J=8.2Hz,1H),4.55(q,J=7.2Hz,2H),3.52–3.39(m,3H ),3.29–3.16(m,2H),2.37–2.23(m,2H),2.22–2.03(m,3H),1.86–1.73(m,4H),1.71–1.43(m,10 H),1.40(d,J=7.2Hz,3H),1.36(d,J=7.1Hz,3H),1.15(t,J=7.6Hz,3H).MS(ESI)m / z=659.2[M+H] + .

[0270] Example 8. Synthesis of N-((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)phenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0271]

[0272] Step 1: At 10°C, LiOH·H₂O (209 mg, 4.97 mmol) was added to a THF (3 mL) / H₂O (1 mL) solution of ethyl 2-(3-fluoro-4-nitrophenyl)propionate (300 mg, 1.24 mmol). The mixture was stirred at this temperature for 4 hours, and then stirred at 25°C for 16 hours. The mixture was acidified to pH 4 with 2N HCl, diluted with EA (30 mL), washed with water (10 mL * 3), dried over Na₂SO₄, and concentrated to give a brown oily crude product of 2-(3-fluoro-4-nitrophenyl)propionate (250 mg, 1.05 mmol, yield: 84.9%). MS (ESI) m / z = 214.1 [M + H] + .

[0273] Step 2: A mixture of 2-(3-fluoro-4-nitrophenyl)propionic acid (250 mg, 1.17 mmol), methyl (2,2,2-trifluoroethyl)amine (351 mg, 2.35 mmol), and EDCI (674 mg, 3.52 mmol) in Pyr (5 mL) was stirred at 25 °C for 16 hours. The mixture was diluted with EA (40 mL), washed with 1N HCl (15 mL x 3) and brine (15 mL x 2), dried over Na2SO4, concentrated, and purified by rapid chromatography (PE / EA = 0–40%) to give 2-(3-fluoro-4-nitrobenzene)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (230 mg, 0.67 mmol, yield: 57.3%) as a yellow oil. MS (ESI) m / z = 309.0 [M+H] + .

[0274] Step 3: A mixture of 2-(3-fluoro-4-nitrophenyl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (230 mg, 0.746 mmol) and Pd / C (23 mg) in THF (10 mL) was stirred at 25 °C under H2 for 16 hours. The mixture was filtered, concentrated, and purified by rapid chromatography (DCM:MA = 0–3%) to give 2-(4-amino-3-fluorophenyl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (200 mg, 0.214 mmol, yield: 91.5%) as a yellow oil. MS (ESI) m / z = 279.0 [M+H] + .

[0275] Step 4: Add DIEA (139.35 mg, 1.08 mmol) to a THF (10 mL) solution of 2-(4-amino-3-fluorophenyl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (150 mg, 0.54 mmol), (S)-{[(tert-butoxy)carbonyl]amino}[4-(difluoromethylene)cyclohexyl]acetic acid (164.6 mg, 0.54 mmol), and HATU (307.5 mg, 0.81 mmol). Stir the reaction mixture at room temperature for 16 hours. Dilute the mixture with water and extract with EA (50 mL x 3). The combined organic layers were washed with brine (150 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (PE / EtOAc = 30-50%) to give tert-butyl ((1S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-(1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)phenyl)amino)-2-oxoethyl)carbamate (208 mg, 0.22 mmol, yield: 40.9%), as a pale yellow solid. MS (ESI) m / z = 588.3 [M + Na] + .

[0276] Step 5: Add TFA (5 mL) to a DCM (10 mL) solution of N-[(S)-[4-(difluoromethylene)cyclohexyl][(2-fluoro-4-{1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl}phenyl)carbamoyl]methyl]tert-butyl carbamate (208 mg, 0.37 mmol). Stir the reaction mixture at room temperature for 3 hours. Concentrate under vacuum and purify by rapid chromatography (DCM / MeOH = 30-50%) to give (2S)-2-(4-((2S)-2-amino-2-(4-difluoromethylene)cyclohexyl)acetamityl)-3-fluorophenyl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (122 mg, 0.26 mmol, yield: 71.3%) as a pale yellow solid. MS (ESI) m / z = 466.3 [M+Na] + .

[0277] Step 6: Add DIEA (67.75 mg, 0.52 mmol) to a DMF (10 mL) solution of (2S)-2-(4-((2S)-2-amino-2-(4-difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (122 mg, 0.26 mmol), 2-isopropylpyrazole-3-carboxylic acid (40.41 mg, 0.26 mmol), and HATU (149.49 mg, 0.39 mmol). Stir the reaction mixture at room temperature for 2 hours. Dilute the mixture with water and extract with EA (50 mL x 3). Wash the combined organic layers with brine (150 mL), dry with Na2SO4, concentrate under vacuum, and purify by rapid chromatography (MeOH-DCM = 0-5%) and preparative TLC (DCM / MeOH = 1 / 20) to obtain Example 8, a white solid. 1 H NMR (400MHz, DMSO-d6) δ9.96 (s, 1H), 8.51 (d, J = 8.1Hz, 1H), 7.76 (dd, J = 16.0, 7.8Hz, 1H), 7.50 (d, J = 1.8Hz, 1H ),7.18–7.01(m,2H),6.95(d,J=1.8Hz,1H),5.45–5.31(m,1H),4.59(t,J=8.2Hz,1H),4.36–4.23(m,1H),4.24– 4.15(m,1H),4.15–3.99(m,1H),2.95(d,J=35.1Hz,3H),2.46–2.36(m,2H),2.00(q,J=10.5Hz,1H),1.92–1.71( m,4H),1.39–1.31(m,6H),1.28(d,J=6.8Hz,3H),1.15(ddd,J=23.8,12.1,3.3Hz,2H).MS(ESI)m / z=602.3[M+H] + .

[0278] Example 12. Synthesis of N-((2R,3S)-3-(4-(((R)-2-cycloheptyl-2-(6-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-2-yl)acetamyl)-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl)propionamide

[0279]

[0280] Step 1: Under an Ar atmosphere, a mixture of 4-bromo-2-nitroaniline (3 g, 13.8 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (3.19 g, 15.1 mmol), Pd(dppf)Cl2 (500 mg, 0.6 mmol), and K2CO3 (5.72 g, 41.4 mmol) in dioxane (30 mL) and H2O (5 mL) was stirred at 95 °C for 16 hours. The mixture was diluted with water (30 mL) and extracted with EA (30 mL x 3). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, concentrated under vacuum, and purified by rapid chromatography (DCM / MeOH = 0-2%) to give an orange solid product (2.2 g, 72%). MS(ESI)m / z = 221.1[M+H] + .

[0281] Step 2: Pd / C (240 mg) was added to a solution of 4-(3,6-dihydro-2H-pyran-4-yl)-2-nitroaniline (2.4 g, 10.9 mmol) in EtOAc (20 mL) and THF (20 mL). The reaction mixture was stirred at room temperature under H2 (balloon) for 16 hours. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under vacuum to give a crude product (2.1 g, crude product) as a yellow solid. 1 H NMR (400MHz, CDCl3) δ6.71–6.63(m,1H),6.63–6.52(m,2H),4.14–4.02(m,2H),3.50(td,J =11.3,3.3Hz,2H),3.29(s,4H),2.60(m,1H),1.83–1.66(m,4H).MS(ESI)m / z=193.1[M+H] + .

[0282] Step 3: A mixture of ethyl 2-cyano-2-cycloheptyl acetate (979.7 mg, 4.68 mmol) and 4-(oxacyclohexyl-4-yl)phenyl-1,2-diamine (600 mg, 3.12 mmol) was stirred at 185 °C for 4 hours. The reaction mixture was concentrated under vacuum and purified by rapid chromatography (PE / EA = 0-35%) to give a yellow solid product (456 mg, 43% yield). MS (ESI) m / z = 338.2 [m+H]+.

[0283] Step 4: A mixture of ethyl 2-cyanoacetate (4791 mg, 42.3 mmol), bromocycloheptane (5 g, 28.2 mmol), and EtONa (3.46 g, 50.8 mmol) in ethanol (100 mL) was stirred at 90 °C for 16 hours. The mixture was concentrated and purified by rapid chromatography (PE:EA = 0–5%) to give a colorless oily product (2.03 g, 6.79 mmol, 24% yield).

[0284] 1 H NMR (400MHz, CDCl3) δ4.27(q,J=7.2Hz,2H),3.42(d,J=5.3Hz,1H),2.31–2.18(m,1H),1.79–1.72(m,4H),1.58–1.46(m,8H),1.32(t,J=7.2Hz,3H).

[0285] Step 5: A solution of 2-cycloheptayl-2-[5-(oxacyclohexyl-4-yl)-3H-1,3-benzodiazol-2-yl]acetonitrile (456 mg, 1.35 mmol) in AcOH (8 mL) and HCl (8 mL) was stirred at 100 °C for 16 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a brown solid product (350 mg, 0.98 mmol, yield 72.9%). MS: (ESI) m / z = 357.1 [M + H] + .

[0286] Steps 6 and 7: A mixture of cycloheptyl[5-(oxacyclohexyl-4-yl)-3H-1,3-benzodiazol-2-yl]acetic acid (120 mg, 0.34 mmol), N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]acrylamide (118 mg, 0.34 mmol), HATU (155 mg, 0.4 mmol), and DIPEA (130 mg, 1 mmol) in THF (10 mL) was stirred at ambient temperature for 16 hours. The reaction mixture was concentrated under vacuum and purified by rapid chromatography (PE / EA = 0-20%) to give a white solid product (30 mg, 0.04 mmol, 12.8% yield). The product was purified by SFC (CHIRALPAK AD-H column, 250 mm * 20 mm, 5 μm) to obtain a white solid, as described in Example 12 (yield 3%). 1HNMR (400MHz, CD3OD) δ7.84(t,J=8.2Hz,1H),7.41(d,J=27.4Hz,2H),7.11(t,J=8.8Hz,2H),7.01(d,J=8.4Hz,1H),4 .88(s,1H),4.02(d,J=12.8Hz,2H),3.88(d,J=10.6Hz,1H),3.62–3.46(m,3H),3.39(d,J=13.3Hz,1H),3.35–3.30(m ,1H),3.10(tt,J=17.9,8.9Hz,2H),2.92–2.78(m,1H),2.58–2.44(m,1H),2.31–2.15(m,4H),1.91(d,J=17.9Hz,4H) ,1.84–1.67(m,6H),1.65–1.34(m,11H),1.29(d,J=6.8Hz,3H),1.09(dd,J=8.1,7.1Hz,3H).MS(ESI)m / z=689.3[M+H] + .

[0287] Using steps similar to those used in Examples 1 to 12, the following compounds were obtained, as shown in Table 1.

[0288] Table 1

[0289]

[0290]

[0291]

[0292]

[0293]

[0294] Example 28. Synthesis of N-((S)-1-cycloheptayl-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-(difluoromethylene)piperidinyl-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0295]

[0296] Step 1: Sodium borohydride (6.2 g, 164 mmol) was added to a methanol (600 mL) solution of 1-(3-fluoro-4-nitrophenyl)ethyl ketone (50 g, 273 mmol) at 0 °C. The mixture was stirred at 0 °C for 1.5 h. The reaction was quenched with ice water (800 mL) and extracted with EA (500 mL * 3). The combined organic layers were washed with brine (500 mL), dried with Na2SO4, and then concentrated under vacuum to give 1-(3-fluoro-4-nitrophenyl)ethyl-1-ol (49 g, 264.6 mmol, yield: 96.9%) as a brown oil. 1 H NMR (400MHz, CDCl3) δ8.05(dd,J=8.3,7.7Hz,1H),7.35–7.27(m,2H),4.99(q,J=6.5Hz,1H),2.07(s,1H),1.52(d,J=6.5Hz,3H).

[0297] Step 2: At 0°C, PPh3 (83.28 g, 317.5 mmol) and tetrabromomethane (105.30 g, 317.5 mmol) were added to a DCM (600 mL) solution of 1-(3-fluoro-4-nitrophenyl)ethanol (49 g, 265 mmol). The mixture was stirred at room temperature for 1.5 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (EtOAc / hexane = 0–10%) to give 4-(1-bromoethyl)-2-fluoro-1-nitrobenzene (53 g, 213.7 mmol, yield: 80.8%) as a pale yellow oil.

[0298] 1 H NMR (400MHz, CDCl3) δ8.10-8.02(m,1H),7.42-7.32(m,2H),5.13(q,J=6.9Hz,1H),2.04(d,J=6.9Hz,3H).

[0299] Step 3: With stirring, sodium hydroxide (12 g, 301 mmol) was added to a DMF (300 mL) solution of (SP-4-4)-[N-[phenyl[2-[[[(1R,2S)-1-(phenylmethyl)-2-pyrrolyl-κN]carbonyl]amino-κN]-phenyl]methylene]glycine(2-)-κN,κO]nickel (30 g, 60.2 mmol)]. Then, a DMF solution (30 mL) of 4-(1-bromoethyl)-2-fluoro-1-nitrobenzene (20.9 g, 84.2 mmol) was added to the solution. The mixture was stirred under nitrogen, and the reaction progress was monitored by TLC. The reaction mixture was quenched with ice water (1200 mL) and extracted with ethyl acetate (600 mL x 3). The organic layers were combined, washed with brine (600 mL x 2), dried over Na₂SO₄, and concentrated under vacuum to obtain the crude product. This crude product was purified by rapid chromatography (elution gradient: EtOAc / hexane = 0-55%) to give compound Example 28-1 (12 g, 18 mmol, yield: 29.9%) as a deep red solid. MS (ESI) m / z = 665.2 [M + H] + .

[0300] Steps 4-5: Add 3NHCl / MeOH = 1:1 (400 mL) to a MeOH solution of compound Example 28-1 (31 g, 46.6 mmol) in 300 mL, and stir the mixture at 70 °C for 10 minutes. Dilute the mixture with water (100 mL) and wash with DCM (400 mL * 3). Then add water, add Na2CO3 to adjust the pH to 10, add THF (500 mL) and Boc2O (108 g, 466 mmol). Stir the mixture further at room temperature for 16 hours. Dilute the mixture with EA (1000 mL) and wash with water (600 mL * 3). The combined liquid layers were acidified to pH 3 with 4M HCl, washed with EtOAc (600 mL * 3), and the combined organic layers were concentrated to give the crude product (2R, 3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-3-fluoro-4-nitrophenyl)butyric acid (11 g, 32.1 mmol, yield: 68.9%), as a pale yellow solid. MS (ESI) m / z = 365.0 [M + Na] + .

[0301] Step 6: Add DIEA (1.1 g, 8.76 mmol) to a DMF (15 mL) solution of (2R,3S)-2-{[(tert-butoxy)carbonyl]amino}-3-(2-chloro-3-fluoro-4-nitrophenyl)butyric acid (1.0 g, 2.92 mmol), 4-(difluoromethylene)piperidine hydrochloride (593 mg, 3.5 mmol), and HATU (1.33 g, 3.5 mmol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (50 mL) and extract with EtOAc (50 mL x 3). The organic layer was washed with brine (50 mL * 2), dried over anhydrous Na₂SO₄, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: EtOAc / hexane = 1 / 1, v / v) to give ((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)carbamate (1.1 g, 2.4 mmol, yield: 82.2%) as a yellow solid. MS (ESI) m / z = 479.8 [M + Na] + .

[0302] Step 7: Add TFA (5 mL) to a DCM (10 mL) solution of ((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)carbamate (900 mg, 1.97 mmol). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the mixture under reduced pressure. Alkalize the residue with a saturated aqueous solution of NaHCO3 and extract with EtOAc (50 mL * 3). Dry the combined organic layers with anhydrous Na2SO4 and concentrate under reduced pressure to give a yellow solid of (2R,3S)-2-amino-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)but-1-one (700 mg, 1.96 mmol, 99.5% yield). MS(ESI)m / z = 358.1 [M+H] + .

[0303] Step 8: At 0°C, trifluoroacetic anhydride (252 mg, 1.20 mmol) was added to a DCM solution of (2R,3S)-2-amino-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)but-1-one (330 mg, 0.92 mmol) and DIEA (239 mg, 1.85 mmol) in 10 mL. The mixture was stirred overnight at room temperature. Quenching was performed with saturated NaHCO3 (20 mL) and extraction was performed with dichloromethane (20 mL x 3). The organic layers were combined, washed with brine (20 mL), and dried over anhydrous sodium sulfate. The mixture was then filtered and concentrated under vacuum to obtain the crude product. The compound N-((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (400 mg, 0.88 mmol, yield: 95.3%) was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-20%) as a white solid. MS (ESI) m / z = 454.0 [M+H] + .

[0304] Step 9: Add Raney nickel (120 mg) and hydrazine hydrate (3 mL) to a 10 mL ethanol solution of N-((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (400 mg, 0.88 mmol). Stir the mixture at room temperature for 4 hours. The mixture was filtered through diatomaceous earth and concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (elution gradient: methanol / dichloromethane, 0-2%) to give compound N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (260 mg, 0.61 mmol, yield: 69.6%) as a pale yellow solid. MS (ESI) m / z = 424.1 [M+H] + .

[0305] Step 10: Stir a mixture of N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (280 mg, 0.66 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetic acid (269 mg, 0.99 mmol), HATU (452 ​​mg, 1.19 mmol), and DIEA (213 mg, 1.65 mmol) in THF (8 mL) overnight at room temperature. The product was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-40%) to give compound ((S)-1-cycloheptyl-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)tert-butyl carbamate (270 mg, 0.40 mmol, yield: 60.4%), as a white solid. MS (ESI) m / z = 699.2 [M+Na] + .

[0306] Step 11: Add TFA (2 mL) to a solution of ((S)-1-cycloheptayl-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)carbamate (270 mg, 0.40 mmol) in dichloromethane (6 mL) and stir the mixture at room temperature for 2 hours. Quench with saturated NaHCO3 (20 mL) and extract with dichloromethane (20 mL * 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: methanol / dichloromethane, 0–2%) to give N-((2R,3S)-3-(4-((S)-2-amino-2-cycloheptylacetamido)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (110 mg, 0.19 mmol, yield: 47.8%) as a grayish-white solid. MS (ESI) m / z = 577.2 [M+H] + .

[0307] Step 12: The mixture of N-((2R,3S)-3-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (110 mg, 0.19 mmol), 2-ethylpyrazole-3-carboxylic acid (35 mg, 0.25 mmol), HATU (116 mg, 0.30 mmol), and DIEA (49 mg, 0.38 mmol) in DMF (4 mL) was stirred at room temperature for 2 hours. It was then diluted with water (40 mL) and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-50%) to give N-((S)-1-cycloheptyl-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (110 mg, 0.16 mmol, yield: 82.6%) as a grayish-white solid. MS (ESI) m / z = 699.2 [M+H] + .

[0308] Step 13: Add lithium hydroxide monohydrate (16 mg, 0.393 mmol) to a solution of N-((S)-1-cycloheptayl-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (110 mg, 0.157 mmol) in THF / water = 1:1 (4 mL), and stir the mixture overnight at room temperature. Then dilute with water (15 mL) and extract with ethyl acetate (10 x 3 mL). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give compound N-((S)-2-((4-(2S,3R)-3-amino-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-cycloheptyl-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (70 mg, 0.116 mmol, yield: 73.8%), as a white solid. MS (ESI) m / z = 603.4 [M+H] + .

[0309] Step 14: At 0°C, triphosgene (16 mg, 0.052 mmol) was added to a solution of N-((S)-2-((4-(2S,3R)-3-amino-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-cycloheptyl-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (70 mg, 0.116 mmol) and DIEA (60 mg, 0.464 mmol) in dichloromethane (5 mL), and the mixture was stirred at room temperature for 10 minutes. Then N-methylcyclopropylamine (17 mg, 0.162 mmol) was added. The mixture was stirred at room temperature for 4 hours. Quenching was performed with saturated NaHCO3 (10 mL), and extraction was performed with dichloromethane (10 mL * 3). The organic layers were combined, washed with brine (20 mL), and dried over anhydrous sodium sulfate. The product was then filtered and concentrated under vacuum to obtain the crude product. It was then subjected to preparative HPLC (HPLC mobile phase: ACN-H₂O (0.1% FA), 35 / 65-75 / 25) to yield compound N-((S)-1-cycloheptyl-2-((4-(2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (41 mg, 0.059 mmol, yield: 50.5%), a white solid. MS (ESI) m / z = 700.5 [M+H] + . 1 H NMR (400MHz, DMSO) δ9.88(s,1H),8.45(d,J=8.6Hz,1H),7.67(t,J=8.3Hz,1H),7.47(d,J=2.0Hz,1H),7.15(dd,J=1 2.0,1.4Hz,1H),7.09–7.03(m,1H),6.99(d,J=2.0Hz,1H),6.18(d,J=9.0Hz,1H),4.79(t,J=8.9Hz,1H),4.60(t,J=8 .5Hz,1H),4.47(q,J=7.1Hz,2H),3.50–3.39(m,2H),3.34(s,1H),3.21-3.08(m,2H),2.74(s,3H),2.56–2.51(m,1H) ,2.14-1.94(m,3H),1.77–1.59(m,5H),1.59–1.32(m,9H),1.31-1.21(m,6H),0.91–0.75(m,2H),0.68-0.55(m,2H).

[0310] Example 29. Synthesis of N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0311]

[0312] Step 1: Add HATU (1.08 g, 2.84 mmol) and DIEA (0.37 mg, 2.84 mmol) to a THF (15 mL) solution of (S)-2-(tert-butoxycarbonyl)amino)-2-((1r,4S)-4-methylcyclohexyl)acetic acid (0.77 g, 2.84 mmol). Stir the mixture at 25 °C for 30 min. Then add N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (1 g, 2.36 mmol). Stir the mixture at room temperature for 24 h. Dilute the mixture with water (20 mL) and extract with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 3 / 2, v / v) to give tert-butyl ((S)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate (1.5 g, 2.22 mmol, yield: 94.1%) as a white solid. MS (ESI) m / z = 699.0 [M+Na] + .

[0313] Step 2: Add TFA (10 mL) to a DCM (10 mL) solution of ((S)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate (1.5 g, 2.22 mmol). Stir the mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL) and adjust the pH to 8 with NaHCO3, then extract with ethyl acetate (50 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give crude N-((2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-methylcyclohexyl)acetamityl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (1 g, 1.74 mmol, yield: 78.4%), as a grayish-white solid. MS (ESI) m / z = 577.3 [M+H] + .

[0314] Step 3: Add HATU (0.79 g, 2.0 mmol) and DIEA (0.29 g, 2.09 mmol) to a DMF (15 mL) solution of 1-ethyl-1H-pyrazole-5-carboxylic acid (0.29 g, 2.09 mmol). Stir the mixture at 25 °C for 30 minutes. Then add N-((2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)-2,2,2-trifluoroacetamide (1 g, 1.74 mmol). Stir the mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL) and extract with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 3 / 1, v / v) to give N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (1.2 g, 1.72 mmol, yield: 98.9%) as a white solid. MS (ESI) m / z = 699.0 [M+H] + .

[0315] Step 4: Add NaBH4 (16.5 mg, 0.43 mmol) to an ethanol (10 mL) solution of N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(2,2,2-trifluoroacetamido)but-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (200 mg, 0.29 mmol). Stir the mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL) and extract with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: DCM / MeOH, 10 / 1, v / v) to give N-((S)-2-((4-(2S,3R)-3-amino-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (100 mg, 0.17 mmol, yield: 58.6%) as a white solid. MS (ESI) m / z = 603.1 [M+H] + .

[0316] Step 5: Add a solution of N-methylcyclopropylamine (7 mg, 0.1 mmol) in DCM dropwise to a solution of N-((S)-2-((4-(2S,3R)-3-amino-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (30 mg, 0.05 mmol), DIEA (12.8 mg, 0.1 mmol), and bis(trichloromethyl) carbonate (8.9 mg, 0.03 mmol) in DCM (10 mL). Stir the reaction mixture at 25 °C for 1 hour. Pour the reaction mixture into water (30 mL) and extract with EtOAc (20 mL * 3). The organic layer was washed with brine (20 mL * 2), dried over anhydrous Na₂SO₄, and concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (Gemini-C18 column, 5 μm silica, 21 mm diameter, 150 mm length) using a mixture of water (containing 0.05% FA) and MeCN with gradually decreasing polarity as eluent (30-70) to give the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylureido)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (11.3 mg, 0.016 mmol, yield: 32.33%) as a white solid. MS(ESI)m / z = 700.4[M+H] + . 1 H NMR(400MHz,CD3OD)δppm 7.72–7.67(m,1H),7.38(s,1H),7.00–6.96(m,2H),6.75(d,J=2.0Hz,1H),6.19–6.1 7(m,1H),4.81–4.73(m,1H),4.43–4.39(m,3H),3.40–3.35(m,1H),3.25–3.20(m,1H ),3.15–3.04(m,2H),2.78(s,3H),2.48–2.46(m,1H),2.01–1.98(m,1H),1.85–1.65 (m,6H),1.45–1.35(m,1H),1.27–0.92(m,9H),0.85–0.71(m,7H),0.66–0.63(m,2H).

[0317] Example 30. Synthesis of N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-methoxy-4-methylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0318]

[0319] Step 1: Add a methanol solution (40 mL, 120 mmol) of 3N HCl to a methanol solution (20 mL) of (2R,3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (4 g, 11.7 mmol). Stir the reaction mixture at 60 °C for 16 hours. Concentrate the mixture under reduced pressure to give crude (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)butyrate methyl ester (3.0 g, 11.7 mmol, yield: 100.0%) as a yellow solid. MS (ESI) m / z = 256.9 [M+H] + .

[0320] Step 2: At 0°C, triphosgene (1.39 g, 4.6 mmol) was added to a DCM (40 mL) solution of methyl (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)butyrate (2 g, 7.8 mmol) and DIEA (4.03 g, 31.2 mmol). The mixture was stirred at 0°C under nitrogen for 10 minutes. A DCM (10 mL) solution of N-methylcyclopropylamine hydrochloride (2.52 g, 23.3 mmol) was added dropwise. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was poured into water (100 mL) and extracted with DCM (50 mL x 3). The organic layer was washed with brine (100 mL x 2), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 50% EtOAc in hexane) to give methyl (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(3-fluoro-4-nitrophenyl)butyrate (1.7 g, 4.8 mmol, yield: 61.54%) as a yellow gel. MS (ESI) m / z = 353.9 [M+H] + .

[0321] Step 3: Add 10% Pd / C (0.51 g, 0.48 mmol) to a solution of (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(3-fluoro-4-nitrophenyl)butyrate (1.7 g, 4.8 mmol) in 30 mL of EtOAc. Stir the reaction mixture at 25 °C for 4 hours under a hydrogen atmosphere. Filter the mixture through a diatomaceous earth mat and concentrate under reduced pressure. Purify the crude product by rapid silica gel chromatography (elution gradient: 0 to 60% hexane solution of EtOAc) to give methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (1.4 g, 4.3 mmol, yield: 89.58%) as a white solid. MS (ESI) m / z = 323.9 [M+H] + .

[0322] Step 4: Add DIEA (0.8 g, 6.2 mmol) to a THF (40 mL) solution of methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-methylureido)butyrate (1.0 g, 3.1 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetic acid (1.42 g, 4.6 mmol), and HATU (1.77 g, 4.6 mmol). Stir the reaction mixture at 25 °C for 16 hours. Pour the reaction mixture into water (50 mL) and extract with EtOAc (50 mL x 3). Wash the organic layer with brine (50 mL x 2), dry with anhydrous Na2SO4, and concentrate under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 70% EtOAc in hexane) to give methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (1.5 g, 2.5 mmol, yield: 80.65%) as a white solid. MS (ESI) m / z = 610.8 [M+H] + .

[0323] Step 5: Add TFA (3 mL) to a solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (1.5 g, 2.5 mmol) in DCM (15 mL). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the mixture under reduced pressure. Alkalize the residue with a saturated aqueous solution of NaHCO3 and extract with EtOAc (50 mL * 3). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under reduced pressure to give a white solid (2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (1.25 g, 2.4 mmol, 96.00% yield). MS (ESI) m / z = 511.3 [M+H] + .

[0324] Step 6: Add DIEA (0.52 g, 4 mmol) to a stirred solution of (2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (1 g, 2 mmol), 2-ethylpyrazole-3-carboxylic acid (0.42 g, 3 mmol), and HATU (1.14 g, 3 mmol) in DMF (20 mL). Stir the reaction mixture at 25 °C for 2 hours. Pour the reaction mixture into water (100 mL) and extract with EtOAc (50 mL x 3). The organic layer was washed with brine (50 mL x 2), dried over anhydrous Na₂SO₄, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 80% EtOAc in hexane) to give methyl (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (1.2 g, 1.9 mmol, yield: 95.0%) as a white solid. MS (ESI) m / z = 633.3 [M+H] + .

[0325] Step 7: Add dropwise a solution of lithium hydroxide monohydrate (160 mg, 0.38 mmol) in water (3 mL) to a THF (10 mL) solution of methyl (2R, 3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (1.2 g, 1.9 mmol). Stir the reaction mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL), acidify with 1 N HCl, and extract with EtOAc (50 mL * 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to give (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-(S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (1.05 g, 1.7 mmol, yield: 89.47%), as a white solid. MS (ESI) m / z = 619.2 [M+H] + .

[0326] Step 8: Add DIEA (12.5 mg, 0.096 mmol) to a DMF (5 mL) solution of (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-(S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (30 mg, 0.048 mmol), 4-methoxy-4-methylpiperidine hydrochloride (12 mg, 0.072 mmol), and HATU (27.5 mg, 0.072 mol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). Wash the organic layer with brine (20 mL * 2), dry with anhydrous Na₂SO₄, and concentrate under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (Gemini-C18 column, 5 μM silica, 21 mm diameter, 150 mm length) using a mixture of water (containing 0.1% FA) and MeCN as eluents (20%–40%) with gradually decreasing polarity, to give the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylureido)-4-(4-methoxy-4-methylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (11 mg, 0.015 mmol, yield: 31.06%) as a white solid. MS (ESI) m / z = 729.8 [M+H]+ . 1 H NMR(400MHz,MeOD)δppm 7.93-7.82(m,1H),7.51(d,J=2.0Hz,1H),7.19–7.08(m,2H),6.89–6.88(m,1H),4.95(d,J=9. 2Hz,1H),4.63–4.53(m,3H),4.10–3.90(m,1H),3.63(d,J=13.2Hz,1H),3.18–3.09(m,5H),2. 90–2.74(m,5H),2.61–2.53(m,3H),2.10–1.95(m,2H),1.92–1.85(m,3H),1.63–1.59(m,2H), 1.39–1.36(m,6H),1.28–1.22(m,2H),1.11(s,1H),0.99–0.81(m,5H),0.75(d,J=3.6Hz,2H).

[0327] Example 31. Synthesis of N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-oxo-4-(4-(1-(trifluoromethyl)cyclopropyl)piperazinyl-1-yl)but-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0328]

[0329] Step 1: Add N-benzyl-2-chloro-N-(2-chloroethyl)ethyl-1-amine hydrochloride (300 mg, 1.12 mmol), 1-(trifluoromethyl)cyclopropylamine (140 mg, 1.12 mol), and DIPEA (2 mL) to a sealed tube. Heat the mixture to 120 °C for 2 days. Cool the reaction to room temperature and concentrate under reduced pressure. Dilute the residue with EtOAc (30 mL) and wash with water (20 mL x 2) and brine (20 mL). Dry the organic layer with sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography (0-60% EtOAc / hexane) to give 1-benzyl-4-(1-(trifluoromethyl)cyclopropyl)piperazine (150 mg, 0.53 mmol, yield: 47.1%) as a yellow gel. MS (ESI) m / z = 285.0 [M+H] + .

[0330] Step 2: 1-Benzyl-4-(1-(trifluoromethyl)cyclopropyl)piperazine (102 mg, 0.359 mmol) and 10% Pd / C (11 mg, 0.1 mmol) were suspended in methanol (7 mL). The reaction mixture was stirred at room temperature under 50 psi hydrogen for 6 hours. The mixture was filtered through a diatomaceous earth mat, and the filtrate was concentrated under reduced pressure to give 1-(1-(trifluoromethyl)cyclopropyl)piperazine (50 mg, 0.26 mmol, yield: 71.7%) as a colorless gel. MS (ESI) m / z = 195.2 [M+H] + .

[0331] Step 3: Add DIEA (12.5 mg, 0.096 mmol) to a DMF (5 mL) solution of (2R, 3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (30 mg, 0.048 mmol), 1-(1-(trifluoromethyl)cyclopropyl)piperazine (14 mg, 0.072 mmol), and HATU (27.5 mg, 0.072 mmol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). Wash the organic layer with brine (20 mL * 2), dry with anhydrous Na2SO4, and concentrate under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (Gemini-C18 column, 5 μM silica, 21 mm diameter, 150 mm length) using a mixture of water (containing 0.1% FA) and MeCN as eluents (20-40) with gradually decreasing polarity, yielding the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylureido)-4-oxo-4-(4-(1-(trifluoromethyl)cyclopropyl)piperazin-1-yl)but-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (18.9 mg, 0.024 mmol, yield: 50.0%) as a white solid. MS (ESI) m / z = 795.0 [M+H] + . 1H NMR(400MHz,CD3OD)δppm 7.93(t,J=8Hz,1H),7.50(d,J=2Hz,1H),7.18–7.08(m,2H),6.88(d,J=2Hz,1H),4.95–4 .88(m,1H),4.67–4.51(m,3H),3.47(br.s,1H),3.29(br.s,2H),3.20–3.09(m,2H),2.9 0(s,3H),2.77–2.66(m,2H),2.64–2.50(m,3H),2.45–2.37(m,1H),2.21–1.85(m,6H),1 .42–1.36(m,6H),1.34–1.24(m,2H),0.97–0.92(m,4H),0.84(s,2H),0.78–0.73(m,2H).

[0332] Example 32. Synthesis of N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-(methyl-d3)ureo)-4-(4-methoxy-4-methylpiperidinyl-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0333]

[0334] Step 1: Add a solution of iodomethane-d3 (2.74 g, 18.9 mmol) in tetrahydrofuran (10 mL) to a THF solution of 2 g (20 mL) containing 2 g (12.6 mmol) of cyclopropyl tert-butyl carbamate and 0.76 g (18.9 mmol) of NaH, stirred at 0 °C for 10 min. Stir the reaction mixture at 25 °C for 16 h. Pour the reaction mixture into water (100 mL) and extract with EtOAc (50 mL x 3). Wash the organic layer with brine (50 mL x 2), dry with anhydrous Na2SO4, and concentrate under vacuum to give cyclopropyl(methyl-d3) tert-butyl carbamate (1.2 g, 6.8 mmol, yield: 53.97%) as a yellow oil. MS (ESI) m / z = 175.0 [M+H] + .

[0335] Step 2: A solution of dioxane (10 mL, 40 mmol) in 4N HCl was added to a solution of 1.2 g (6.8 mmol) of tert-butyl cyclopropylcarbamate in EtOAc (10 mL). The reaction mixture was stirred at 25 °C for 16 hours. The mixture was concentrated under reduced pressure to give N-(methyl-d3)cyclopropylamine hydrochloride (0.5 g, 6.7 mmol, yield: 97.1%) as a yellow gel. 1 H NMR(400MHz, DMSO-d6)δppm 9.18(s,2H),2.66–2.63(m,1H),0.85–0.83(m,2H),0.73–0.70(m,2H).

[0336] Step 3: At 0°C, triphosgene (86 mg, 0.31 mmol) was added to a DCM solution of methyl (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)butyrate (100 mg, 0.39 mmol) and DIEA (100 mg, 0.78 mmol) in 10 mL. The mixture was stirred at 0°C under nitrogen for 10 minutes. A DCM solution of N-(methyl-d3)cyclopropylamine hydrochloride (80 mg, 0.78 mmol) in 2 mL was added dropwise. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was poured into water (20 mL) and extracted with DCM (20 mL x 3). The organic layer was washed with brine (20 mL x 2), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 50% EtOAc in hexane) to give methyl (2R,3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(3-fluoro-4-nitrophenyl)butyrate (110 mg, 0.31 mmol, yield: 79.5%) as a yellow gel. MS (ESI) m / z = 356.9 [M+H] + .

[0337] Step 4: Add 10% Pd / C (65 mg, 0.06 mmol) to a solution of (2R,3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(3-fluoro-4-nitrophenyl)butyrate (110 mg, 0.31 mmol) in 10 mL of EtOAc. Stir the reaction mixture at 25 °C for 3 hours under a hydrogen atmosphere. Filter the mixture through a diatomaceous earth mat, and concentrate the filtrate under reduced pressure to give the crude product. Purify the crude product by rapid silica gel chromatography (elution gradient: 0 to 60% hexane solution of EtOAc) to give methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureo)butyrate (80 mg, 0.245 mmol, yield: 79.4%) as a white solid. MS(ESI)m / z = 326.9[M+H] + .

[0338] Step 5: Add DIEA (63 mg, 0.49 mmol) to a solution of (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureo)butyrate (80 mg, 0.245 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetic acid (112 mg, 0.367 mmol), and HATU (140 mg, 0.367 mmol) in THF (10 mL). Stir the reaction mixture at 25 °C for 16 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). Wash the organic layer with brine (20 mL * 2), dry with anhydrous Na2SO4, and concentrate under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 70% EtOAc in hexane) to give methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureoyl)butyrate (110 mg, 0.179 mmol, yield: 90.24%) as a white solid. MS (ESI) m / z = 614.3 [M+H] + .

[0339] Step 6: Add TFA (2 mL) to a DCM (6 mL) solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureo)butyrate (110 mg, 0.179 mmol). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the mixture under reduced pressure. Alkalize the residue with a saturated aqueous solution of NaHCO3 and extract with EtOAc (20 mL * 3). The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under reduced pressure to give methyl (2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureoyl)butyrate (80 mg, 0.156 mmol, 87.04% yield, white solid). MS (ESI) m / z = 514.2 [M+H] + .

[0340] Step 7: Add DIEA (40 mg, 0.31 mmol) to a DMF (5 mL) solution of (2R, 3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-(methyl-d3)ureoyl)butyrate (80 mg, 0.155 mmol), 2-ethylpyrazole-3-carboxylic acid (32.7 mg, 0.233 mmol), and HATU (88.8 mg, 0.231 mmol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). The organic layer was washed with brine (20 mL x 2), dried over anhydrous Na₂SO₄, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 80% hexane solution of EtOAc) to give methyl (2R, 3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (90 mg, 0.14 mmol, yield: 90.89%) as a white solid. MS (ESI) m / z = 636.3 [M+H] + .

[0341] Step 8: Add dropwise a solution of lithium hydroxide monohydrate (12 mg, 0.28 mmol) in water (2 mL) to a THF (5 mL) solution of ((2R,3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (90 mg, 0.14 mmol). Stir the reaction mixture at 25 °C for 2 hours. Dilute the mixture with water (20 mL) and add 1 N... Acidified with HCl and extracted with EtOAc (20 mL * 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure to give (2R, 3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (75 mg, 0.12 mmol, yield: 85.17%), as a white solid. MS (ESI) m / z = 621.8 [M + H] + .

[0342] Step 9: Add DIEA (12.5 mg, 0.096 mmol) to a DMF (5 mL) solution of (2R,3S)-2-(3-cyclopropyl-3-(methyl-d3)ureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (30 mg, 0.048 mmol), 4-methoxy-4-methylpiperidine hydrochloride (12 mg, 0.072 mmol), and HATU (27.5 mg, 0.072 mol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). Wash the organic layer with brine (20 mL * 2), dry with anhydrous Na2SO4, and concentrate under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (Gemini-C18 column, 5 μM silica, 21 mm diameter, 150 mm length) using a mixture of water (containing 0.1% FA) and MeCN as eluents (20-40) with gradually decreasing polarity, to give the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-(methyl-d3)ureido)-4-(4-methoxy-4-methylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (11 mg, 0.015 mmol, yield: 31.06%) as a white solid. MS (ESI) m / z = 732.8 [M+H] + . 1 H NMR(400MHz,CD3OD)δppm 7.77–7.69(m,1H),7.39(s,1H),7.04–6.95(m,2H),6.78–6.76(m,1H),4.83–4.8 0(m,1H),4.47–4.41(m,3H),3.92–3.80(m,1H),3.49–3.48(m,1H),3.05–2.98(m ,4H),2.65–2.55(m,2H),2.51–2.41(m,3H),1.92–1.77(m,5H),1.53–1.38(m,2H ),1.33–0.98(m,10H),0.74–0.68(m,4H),0.64–0.54(m,2H),0.25–0.17(m,1H).

[0343] Using a procedure similar to that used in Examples 1 to 12 and Examples 28 to 32, the following compounds were obtained. The compounds are shown in Table 2.

[0344] Table 2

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353]

[0354]

[0355]

[0356]

[0357]

[0358]

[0359]

[0360]

[0361]

[0362]

[0363]

[0364]

[0365]

[0366]

[0367]

[0368]

[0369]

[0370]

[0371]

[0372]

[0373]

[0374]

[0375]

[0376]

[0377]

[0378]

[0379]

[0380]

[0381]

[0382]

[0383]

[0384]

[0385]

[0386]

[0387]

[0388]

[0389]

[0390]

[0391]

[0392]

[0393]

[0394]

[0395]

[0396]

[0397]

[0398]

[0399]

[0400] Example 241:

[0401] N-((S)-1-Cycloheptyl-2-((4-(((2S,3R)-3-(3-Cyclopropyl-3-methylureo)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0402]

[0403] Step 1: To a THF (10 mL) solution of methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (100 mg, 0.31 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetic acid (252.72 mg, 0.93 mmol), DIEA (119.70 mg, 0.93 mmol), and HATU (129.29 mg, 0.34 mmol) were added. The mixture was stirred at 40 °C for 16 hours. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 1, v / v) to give methyl (2R, 3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (80 mg, 0.14 mmol, yield: 44.86%) as a white solid. MS (ESI) m / z = 576.9 [M+H] + .

[0404] Step 2: Add TFA (2 mL) to a DCM (5 mL) solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (80 mg, 0.14 mmol) and stir the mixture at 25 °C for 1 hour. Dilute the mixture with water (10 mL). Adjust the pH of the mixture to 8 with NaHCO3 and extract with ethyl acetate (10 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give crude product (2R,3S)-3-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (50 mg, 0.10 mmol, yield: 75.63%), as a white solid. MS (ESI) m / z = 477.0 [M+H] + .

[0405] Step 3: To a DMF (10 mL) solution of methyl (2R,3S)-3-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (50 mg, 0.10 mmol), 1-isopropyl-1H-pyrazole-5-carboxylic acid (13.53 mg, 0.16 mmol), DIEA (40.60 mg, 0.31 mmol), and HATU (43.85 mg, 0.12 mmol) were added, and the mixture was stirred at 25 °C for 1 hour. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 10, v / v) to give methyl (2R, 3S)-3-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (50 mg, 0.082 mmol, yield: 77.78%) as a white solid. MS (ESI) m / z = 612.8 [M+H] + .

[0406] Step 4: Add a 5 mL solution of LiOH (17.12 mg, 0.41 mmol) in water to a 10 mL THF solution of (2R, 3S)-3-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (50 mg, 0.082 mmol). Stir the mixture at 25 °C for 16 hours. Dilute the mixture with 20 mL of water. Adjust the pH of the mixture to 3 with 1 N HCl and extract with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product (2R,3S)-3-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyric acid (35 mg, 0.058 mmol, yield: 71.63%), as a white solid. MS (ESI) m / z = 599.3 [M+H] + .

[0407] Step 5: Add 4-(difluorophenyl)piperidine hydrochloride (10.20 mg, 0.060 mmol), DIEA (19.39 mg, 0.15 mmol), and HATU (20.94 mg, 0.06 mmol) to a DMF (5 mL) solution of (2R, 3S)-3-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyric acid (30 mg, 0.050 mmol) and HATU (20.94 mg, 0.06 mmol). Stir the mixture at 25 °C for 2 hours. Dilute the mixture with water (10 mL) and extract with ethyl acetate (10 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by general-purpose (Genal) preparative HPLC (elution gradient: acetonitrile / water (0.1% FA)) to give the title compound N-((S)-1-cycloheptyl-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide (6 mg, 8.4 mmol, yield: 16.8%). MS (ESI) m / z = 714.3 [M+H] + . 1H NMR(400MHz,MeOD)δppm 7.68(t,J=8.2Hz,1H),7.41(d,J=2.0Hz,1H),7.06-7.00(m,2H),6.67(d,J=2.0Hz,1H),5.33–5.19(m ,1H),4.79(s,1H),4.51(d,J=8.0Hz,1H),3.45-3.36(m,1H),3.32–3.23(m,2H),3.21-3.16(m,1H),3 .10-3.04(m,1H),2.78(s,3H),2.54–2.46(m,1H),2.06-1.97(m,3H),1.79–1.59(m,5H),1.59–1.39( m,7H),1.38-1.27(m,7H),1.28(d,J=7.2Hz,3H),1.19(s,1H),0.85-0.80(m,2H),0.68–0.59(m,2H).

[0408] Example 242:

[0409] N-((S)-1,1-Dicyclobutyl-3-((4-((2S,3R)-3-(3-cyclopropyl-3-methylureo)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobutyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-ethyl-1H-pyrazole-5-carboxamide

[0410]

[0411] Step 1: A mixture of (S)-2-((tert-Butoxycarbonyl)amino)-3,3-dicyclobutylpropionic acid (65 mg, 0.22 mmol), methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (47 mg, 0.15 mmol), HATU (110 mg, 0.29 mmol), and N,N-diisopropylethylamine (56 mg, 0.44 mmol) in DMF (4 mL) was stirred overnight at room temperature. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: PE / EA, 1 / 1, v / v) to give methyl (2R, 3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dicyclobutylpropionamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (40 mg, 0.066 mmol, yield: 45.7%) as a brown solid. MS (ESI) m / z = 603.3 [M+H]+ .

[0412] Step 2: Add TFA (0.5 mL) to a DCM (2 mL) solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-3,3-dicyclobutylpropionamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (40 mg, 0.066 mmol) and stir the mixture at room temperature for 2 hours. Adjust the pH of the mixture to 8-9 with sodium bicarbonate, and then extract with DCM (10 mL * 3). The organic layers were combined, dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to give methyl (2R,3S)-3-(4-((S)-2-amino-3,3-dicyclobutylpropionamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (33 mg, 0.0657 mmol, yield: 99.0%), a light brown solid. MS (ESI) m / z = 503.2 [M+H] + .

[0413] Step 3: The mixture of (2R,3S)-3-(4-((S)-2-amino-3,3-dicyclobutylpropamido)-3-fluorophenyl)-2-(3-cyclopropyl-3-methylurea)butyrate (33 mg, 0.066 mmol), 2-ethylpyrazole-3-carboxylic acid (12 mg, 0.085 mmol), HATU (45 mg, 0.118 mmol), and N,N-diisopropylethylamine (21 mg, 0.164 mmol) in DMF (3 mL) was stirred at room temperature for 2 hours. It was then diluted with water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: PE / EA, 1 / 1, v / v) to give methyl (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-3,3-dicyclobutyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)butyrate (22 mg, 0.035 mmol, yield: 53.6%) as a pale yellow solid. MS (ESI) m / z = 625.2 [M+H] + .

[0414] Step 4: Add LiOH·H₂O (3 mg, 0.070 mmol) to a THF / water solution of methyl (2R, 3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-3,3-dicyclobutyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)butyrate (22 mg, 0.035 mmol) in 1:1 (3 mL) of THF / water and stir the mixture overnight at room temperature. Dilute with water (10 mL) and wash with EtOAc (5 mL). Adjust the aqueous phase to pH 3-4 with 1 N HCl and extract with EtOAc (10 mL x 3). The organic layers were combined, washed with brine, dried over anhydrous sodium sulfate, and concentrated under vacuum to give (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-3,3-dicyclobutyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)butyric acid (20 mg, 0.033 mmol, yield: 94.3%), as a pale yellow solid. MS (ESI) m / z = 611.3 [M+H] + .

[0415] Step 5: The mixture of (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-3,3-dicyclobutyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)propamido)-3-fluorophenyl)butyric acid (10 mg, 0.016 mmol), 4-(difluoromethylene)piperidine (4 mg, 0.026 mmol), HATU (12 mg, 0.033 mmol), and N,N-diisopropylethylamine (8 mg, 0.065 mmol) in DMF (2 mL) was stirred at room temperature for 2 hours. It was then diluted with water (20 mL) and extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (mobile phase: ACN-H₂O (0.1% FA), 40 / 60-80 / 20) to give compound N-((S)-1,1-dicyclobutyl-3-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxobutyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-ethyl-1H-pyrazole-5-carboxamide (2.2 mg, 0.003 mmol, yield: 18.3%) as a white solid. MS (ESI) m / z = 726.3 [M+H] + . 1H NMR (400MHz, CD3OD) δ7.86–7.77(m,1H),7.55–7.49(m,1H),7.21–7.08(m,2H),6.86(s,1H),6.29(d ,J=8.8Hz,1H),4.69(d,J=8.3Hz,1H),4.62–4.49(m,3H),3.52–3.45(m,1H),3.43–3.36(m,2H),3.22 –3.13(m,1H),2.90(s,3H),2.65–2.57(m,1H),2.32–2.24(m,1H),2.16–2.06(m,3H),2.01–1.91(m, 3H),1.84–1.75(m,4H),1.72–1.53(m,8H),1.44–1.38(m,6H),0.97–0.91(m,2H),0.79–0.71(m,2H).

[0416] Examples 243a and 243b:

[0417] N-((S)-1-cycloheptyl-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(propamido-2,2,3,3,3-d5)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0418]

[0419] Step 1: Add HATU (143.00 mg, 0.38 mmol) and DIEA (49.11 mg, 0.38 mmol) to a THF (20 mL) solution of (S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetic acid (103.05 mg, 0.38 mmol). Stir the mixture at 25 °C for 30 min. Then add methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyrate (90 mg, 0.31 mmol). Stir the mixture at 40 °C for 24 h. Dilute the mixture with water (20 mL) and extract with ethyl acetate (20 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: PE / EA = 3 / 2, v / v) to give methyl (2R, 3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetamyl)-3-fluorophenyl)-2-(propamido-2,2,3,3-d5)butyrate (120 mg, 0.22 mmol, yield: 71.0%) as a white solid. MS (ESI) m / z = 541.0 [M+H] + .

[0420] Step 2: Add 2 mL of TFA to a 4 mL solution of (2R,3S)-3-(4-((S)-2-((tert-Butoxycarbonyl)amino)-2-cycloheptylacetamyl)-3-fluorophenyl)-2-(propamido-2,2,3,3-d5)butyrate (120 mg, 0.22 mmol) in DCM. Stir the mixture at 25 °C for 1 hour. Concentrate the mixture under vacuum to obtain crude (2R,3S)-3-(4-((S)-2-amino-2-cycloheptylacetamyl)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyrate (90 mg, 0.20 mmol, yield: 90.9%) as a yellow oil. MS (ESI) m / z = 441.1 [M+H] + .

[0421] Step 3: Add HATU (91.26 mg, 0.24 mmol) and DIEA (31.02 mg, 0.24 mmol) to a DMF (5 mL) solution of 1-ethyl-1H-pyrazole-5-carboxylic acid (33.61 mg, 0.24 mmol). Stir the mixture at 25 °C for 30 minutes. Then add methyl (2R,3S)-3-(4-((S)-2-amino-2-cycloheptylacetamido)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyrate (90 mg, 0.20 mmol). Stir the mixture at 25 °C for 1 hour. Dilute the mixture with water (20 mL) and extract with ethyl acetate (20 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: DCM / MeOH = 10 / 1, v / v) to give methyl (2R,3S)-3-(4-((S)-2-cycloheptyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyrate (90 mg, 0.16 mmol, yield: 80.0%) as a white solid. MS (ESI) m / z = 563.0 [M+H] + .

[0422] Step 4: Add a solution of methyl (2R,3S)-3-(4-((S)-2-cycloheptyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyrate (90 mg, 0.16 mmol) in THF (4 mL) to a solution of LiOH (7.68 mg, 0.32 mmol) in H₂O (2 mL). Stir the mixture at 25 °C for 16 hours. Adjust the pH of the mixture to 4 with 1 M HCl. Extract the mixture with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product (2R,3S)-3-(4-((S)-2-cycloheptyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyric acid (60 mg, 0.11 mmol, yield: 68.8%), as a white solid. MS (ESI) m / z = 549.0 [M+H] + .

[0423] Step 5: Add HATU (25.10 mg, 0.066 mmol) and DIEA (8.53 mg, 0.66 mmol) to a DMF (5 mL) solution of (2R, 3S)-3-(4-((S)-2-cycloheptyl-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(propamido-2,2,3,3,3-d5)butyric acid (30 mg, 0.055 mmol). Stir the mixture at 25 °C for 30 min. Then add 4-(difluoromethylene)piperidine hydrochloride (8.78 mg, 0.066 mmol). Stir the mixture at 25 °C for 1 h. Extract the mixture with ethyl acetate (10 mL * 3). Combine the organic layers, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to give the crude product, which is purified by SFC to obtain the title compound.

[0424] Compound 243a: N-((S)-1-cycloheptyl-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(propamido-2,2,3,3,3-d5)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (4 mg, 0.006 mmol, yield: 10.9%), is a white solid.

[0425] 1 H NMR (400MHz, MeOD) δppm 7.74(t,J=8.3Hz,1H),7.46(d,J=1.9Hz,1H),7.09(t,J=9.1Hz,2H),6.81(d ,J=1.9Hz,1H),5.02–4.89(m,1H),4.58(d,J=8.1Hz,1H),4.50(q,J=7.1Hz, 2H),3.29–3.36(m,3H),3.23–3.14(m,2H),2.19–1.98(m,4H),1.82–1.79(m ,1H),1.75–1.70(m,3H),1.58–1.43(m,7H),1.35–1.30(m,6H),1.26(s,2H).

[0426] MS(ESI)m / z = 663.9[M+H] + .

[0427] Compound 243b: N-((S)-1-cycloheptyl-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-(propamido-2,2,3,3,3-d5)but-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (4 mg, 0.006 mmol, yield: 10.9%), is a solid.

[0428] 1 H NMR(400MHz,MeOD)δppm 7.71(t,J=8.2Hz,1H),7.46(d,J=2.1Hz,1H),7.11(dd,J=11.9,1.7Hz,1H),7.06(d,J=8.4Hz,1 H),6.82(d,J=2.1Hz,1H),5.07(d,J=9.9Hz,1H),4.59(d,J=8.1Hz,1H),4.50(q,J=7.2Hz,2H),3 .91–3.78(m,2H),3.63–3.54(m,1H),3.43–3.37(m,1H),3.25–3.21(m,1H),2.27–2.13(m,4H),1 .85–1.66(m,4H),1.60–1.42(m,7H),1.35(t,J=7.2Hz,3H),1.26(s,2H),1.21(d,J=7.0Hz,3H).

[0429] MS(ESI)m / z = 663.9[M+H] + .

[0430] Example 244:

[0431] Synthesis of N-((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-((S)-2-(1-isopropyl-1H-pyrazole-5-carboxamido)-2-((1r,4S)-4-methylcyclohexyl)acetamyl)phenyl)-1-oxobut-2-yl)-1,2,3-thiadiazole-5-carboxamide

[0432]

[0433] Step 1: A mixture of (2R,3S)-2-amino-1-[4-(difluoromethylene)piperidin-1-yl]-3-(3-fluoro-4-nitrophenyl)but-1-one (160 mg, 0.45 mmol), 1,2,3-thiadiazole-5-carboxylic acid (58 mg, 0.45 mol), HATU (340 mg, 0.896 mmol), and DIEA (174 mg, 1.34 mmol) in THF (6 mL) was stirred at 25 °C for 16 hours. The mixture was diluted with EtOAc (60 mL), washed with water (20 mL x 3) and brine (20 mL), dried over Na2SO4, concentrated, and purified by rapid chromatography (MeOH:DCM = 0–3%) to give a yellow solid product (120 mg, 0.20 mmol, 45.6% yield). MS (ESI) m / z = 470.0 [M+H] + .

[0434] Step 2: To a solution of N-[(2R,3S)-1-[4-(difluoromethylene)piperidin-1-yl]-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl]-1,2,3-thiadiazole-5-carboxamide (150 mg, 0.32 mmol) in ethanol (5 mL), Raney nickel (1 mL) was added, followed by hydrazine hydrate (1 mL). The mixture was stirred at room temperature for 1 hour. The mixture was filtered, the filtrate was concentrated under vacuum, and then purified by preparative TLC (MeOH:DCM = 3%) to give a yellow oily product (22 mg, 0.047 mmol, 14.9% yield). MS (ESI) m / z = 440.1 [M+H] + .

[0435] Step 3: A mixture of N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]-1,2,3-thiadiazole-5-carboxamide (22 mg, 0.05 mmol), (S)-{[(tert-butoxy)carbonyl]amino}(4-methylcyclohexyl)acetic acid (14 mg, 0.05 mmol), HATU (38 mg, 0.10 mmol), and DIEA (19 mg, 0.15 mmol) in THF (2 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by preparative TLC (EtOAc:hexane = 60%) to give a yellow oily product (15 mg, 0.011 mmol, 21.5% yield). MS (ESI) m / z = 593.2 [M+H-Boc] + .

[0436] Step 4: The mixture of N-[(S)-({4-[(2S,3R)-4-[4-(difluoromethylene)piperidin-1-yl]-4-oxo-3-(1,2,3-thiadiazol-5-ylcarbamate)but-2-yl]-2-fluorophenyl}carbamoyl)(4-methylcyclohexyl)methyl]tert-butyl carbamate (15 mg, 0.022 mmol) in TFA (0.2 mL) / DCM (0.2 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily product (12 mg, crude product). MS (ESI) m / z = 593.3 [M+H] + .

[0437] Step 5: The mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-(4-methylcyclohexyl)acetamyl]-3-fluorophenyl}-1-[4-(difluoromethylene)pyridin-1-yl]-1-oxobut-2-yl]-1,2,3-thiadiazole-5-carboxamide (12 mg, 0.020 mmol), 2-isopropylpyrazole-3-carboxylic acid (4 mg, 0.024 mmol), HATU (15 mg, 0.040 mmol), and DIEA (8 mg, 0.61 mmol) in THF (1 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by preparative HPLC (Gemini-C18 150*21.2mm 5um:ACN--H2O(0.1%FA):55-85) to give a white solid product (5.1 mg, 0.0069 mmol, 34.1% yield).

[0438] 1 HNMR (400MHz, CD3OD) δ9.26 (s, 1H), 7.83-7.79 (m, 1H), 7.50 (d, J = 2.0Hz, 1H), 7.19-7.14 (m, 2H) ,6.78(d,J=2.0Hz,1H),5.40-5.33(m,1H),5.13(d,J=10.5Hz,1H),4.50(d,J=8.4Hz,1H),3.52– 3.32(m,4H),3.26–3.18(m,1H),2.18–2.03(m,2H),1.93–1.72(m,5H),1.61–1.52(m,1H),1.47- 1.39(m,8H),1.34–1.29(m,2H),1.25–1.11(m,2H),1.03–0.82(m,6H).MS(ESI)m / z=729.3[M+H] + .

[0439] Example 245:

[0440] Synthesis of N-((2R,3S)-3-(4-((S)-2-(2-(3-cyanophenyl)-2,2-difluoroacetamyl)-2-((1r,4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide

[0441]

[0442] Step 1: Add HOBT (118 mg, 0.88 mmol), EDCI (168 mg, 0.88 mmol), and NMM (236 mg, 2.34 mmol) to a DMF (2 mL) solution of compound (2R, 3S)-2-((tert-butoxycarbonyl)amino)-3-(3-fluoro-4-nitrophenyl)butyric acid (200 mg, 0.58 mmol). Stir the mixture at room temperature for 16 hours. Dilute the mixture with EtOAc (30 mL) and wash with water (20 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by rapid chromatography (elution gradient: EtOAc-hexane, 0-20%) to give ((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)carbamate (200 mg, 0.44 mmol, yield: 75.9%) as a yellow solid. MS: (ESI) m / z = 402.0 [M+H-56] + .

[0443] Step 2: Add TFA (2 mL) to a DCM (1 mL) solution of compound ((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)carbamate (200 mg, 0.44 mmol). Stir the mixture at room temperature for 2 hours. Adjust the pH of the mixture to 7 with NaOH (10 w%). Dilute the mixture with DCM (20 mL) and wash with water (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by rapid chromatography (elution gradient: EtOAc-hexane = 0-20%) to give (2R,3S)-2-amino-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)but-1-one (120 mg, 0.33 mmol, yield: 76.3%) as a yellow solid. MS: (ESI) m / z = 358.2 [M+H] + .

[0444] Step 3: To a THF (2 mL) solution of (2R,3S)-2-amino-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)but-1-one (120 mg, 0.33 mmol), propionyl chloride (51.79 mg, 0.56 mmol) and DIEA (72.34 mg, 0.56 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated to give a crude product, which was purified by rapid chromatography (elution gradient: EtOAc-hexane, 0-50%) to give N-((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)propionamide (100 mg, 0.24 mmol, yield: 72.7%) as a white solid. MS: (ESI)m / z = 414.1 [M+H] + .

[0445] Step 4: N₂H₄·H₂O (30 mg, 0.96 mmol) and Raney nickel were added to a 2 mL ethanol solution of N-((2R,3S)-1-(4-(difluoromethylene)piperidin-1-yl)-3-(3-fluoro-4-nitrophenyl)-1-oxobut-2-yl)propionamide (100 mg, 0.24 mmol). The reaction mixture was stirred at room temperature for 2 hours. The mixture was filtered, concentrated under vacuum, and purified by rapid chromatography (elution gradient: ethyl EtOAc-hexane, 0–20%) to give a yellow oily N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (70 mg, 0.18 mmol, 75.0% yield). MS: (ESI) m / z = 384.3 [M+H] + .

[0446] Step 5: A mixture of N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (70 mg, 0.18 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-((1r,4S)-4-methylcyclohexyl)acetic acid (50 mg, 0.18 mmol), HATU (104 mg, 0.28 mmol), and DIEA (60 mg, 0.46 mmol) in THF (2 mL) was stirred at room temperature for 48 hours. The mixture was diluted with water (20 mL) and extracted with EA (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and purified by rapid chromatography (elution gradient: EtOAc / hexane = 0-50%) to give compound ((S)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate tert-butyl ester (70 mg, 0.11 mmol, yield: 61.1%) as a yellow solid. MS (ESI) m / z = 637.1 [M+H] + .

[0447] Step 6: A solution of ((S)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)carbamate tert-butyl ester (70 mg, 0.11 mmol) in DCM / HCl / dioxane was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum to give compound N-((2R,3S)-3-(4-(((S)-2-amino-2-((1r,4S)-4-methylcyclohexyl)acetamitin)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobutyl-2-yl)propionamide (40 mg, 0.07 mmol, yield: 63.6%). MS(ESI)m / z = 537.1 [M+H] + .

[0448] Step 7: Add 2-(3-cyanophenyl)-2,2-difluoroacetic acid (10 mg, 0.05 mol), HATU (28 mg, 0.07 mmol), and DIEA (13 mg, 0.1 mmol) to a THF (3 mL) solution of N-((2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-methylcyclohexyl)acetamityl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (30 mg, 0.05 mmol). Stir the mixture at room temperature for 16 hours. Dilute the mixture with DCM (20 mL) and wash with water (20 mL x 2) and brine (20 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a crude product. This crude product was purified by preparative TLC (EtOAc / hexane = 1 / 1, v / v) to give N-((2R,3S)-3-(4-((S)-2-(2-(3-cyanophenyl)-2,2-difluoroacetamyl)-2-((1r,4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (3.2 mg, 0.004 mmol, yield: 8.0%), as a white solid. MS (ESI) m / z = 716.4 [M+H] + .

[0449] 1 H NMR (400MHz, CD3OD) δ8.05(s,1H),7.95(dd,J=17.2,7.8Hz,2H),7.73(m,2H),7.16-7.08(m,2H),4.98(d,J=10.3Hz,1H),4.41(d,J=8.7Hz,1 H),3.44(m,4H),3.20(m,1H),2.28(m,4H),2.09(s,3H),1.75(m,5H),1.61(s,1H),1.36(d,J=7.0Hz,3H),1.15(t,J=7.6Hz,3H),0.93(m,6H).

[0450] Examples 246a and 246b:

[0451] Synthesis of N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0452]

[0453] Step 1: Add HATU (133.08 mg, 0.42 mmol) and DIEA (54.28 mg, 0.42 mmol) to a THF (5 mL) solution of methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-propamidobutyrate (100 mg, 0.35 mmol). Stir the mixture at 25 °C for 30 min. Then add (S)-2-((tert-butoxycarbonyl)amino)-2-((1r,4S)-4-methylcyclohexyl)acetic acid (111.11 mg, 0.41 mmol). Stir the mixture at 40 °C for 16 h. Dilute the mixture with water (20 mL) and extract with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain a crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate = 3 / 2, v / v) to give methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-((1r,4S)-4-methylcyclohexyl)acetamito)-3-fluorophenyl)-2-propamidobutyrate (140 mg, 0.26 mmol, yield: 74.8%) as a white solid.

[0454] MS(ESI)m / z = 535.9 [M+H] + .

[0455] Step 2: Add TFA (1 mL) to a DCM (1 mL) solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-((1r,4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-2-propamidobutyrate (100 mg, 0.19 mmol). Stir the mixture at 25 °C for 1 hour. Adjust the pH of the mixture to 8 with NaHCO3 and extract with ethyl acetate (30 mL * 3). Combine the organic layers, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to give crude product (2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-2-propamidobutyrate (70 mg, 0.16 mmol, yield: 84.2%), as a yellow gel.

[0456] MS(ESI)m / z = 436.0 [M+H] + .

[0457] Step 3: Add HATU (73.00 mg, 0.19 mmol) and DIEA (24.56 mg, 1.84 mmol) to a DMF (5 mL) solution of methyl (2R, 3S)-3-(4-((S)-2-amino-2-((1r, 4S)-4-methylcyclohexyl)acetamyl)-3-fluorophenyl)-2-propamidobutyrate (70 mg, 0.16 mmol). Stir the mixture at 25 °C for 30 min. Then add 1-ethyl-1H-pyrazole-5-carboxylic acid (26.6 mg, 0.19 mmol). Stir the mixture at 25 °C for 1 h. Dilute the mixture with water (20 mL) and extract with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: DCM / MeOH = 10 / 1, v / v) to give methyl (2R,3S)-3-(4-((S)-2-(1-ethyl-1H-pyrazole-5-carboxamido)-2-((1r,4S)-4-methylcyclohexyl)acetamito)-3-fluorophenyl)-2-propamidobutyrate (83.5 mg, 0.15 mmol, yield: 97.7%) as a white solid. MS (ESI) m / z = 558.0 [M+H] + .

[0458] Step 4: Add a solution of LiOH (6.88 mg, 0.29 mmol) in H₂O (1 mL) to a THF (2 mL) solution of (2R, 3S)-3-(4-((S)-2-(1-ethyl-1H-pyrazole-5-carboxamido)-2-((1r, 4S)-4-methylcyclohexyl)acetamitoyl)-3-fluorophenyl)-2-propamidobutyrate (80 mg, 0.14 mmol). Stir the mixture at 25 °C for 16 hours. Adjust the pH of the mixture to 4 with 1 N HCl and extract with ethyl acetate (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (2R,3S)-3-(4-((S)-2-(1-ethyl-1H-pyrazole-5-carboxamido)-2-((1r,4S)-4-methylcyclohexyl)acetamitoyl)-3-fluorophenyl)-2-propamidobutyric acid (70.6 mg, 0.13 mmol, yield: 92.9%), as a white solid. MS (ESI) m / z = 544.1 [M+H] + .

[0459] Step 5: Add HATU (25.21 mg, 0.066 mmol) and DIEA (8.53 mg, 0.066 mol) to a DMF (5 mL) solution of (2R, 3S)-3-(4-((S)-2-(1-ethyl-1H-pyrazole-5-carboxamido)-2-((1r, 4S)-4-methylcyclohexyl)acetamido)-3-fluorophenyl)-2-propamidobutyric acid (30 mg, 0.055 mmol). Stir the mixture at 25 °C for 30 min. Then add 4-(difluoromethylene)piperidine (8.78 mg, 0.066 mmol). Stir the mixture at 25 °C for 1 h. Dilute the mixture with water (10 mL) and extract with ethyl acetate (10 mL * 3). Combine the organic layers, dry with anhydrous sodium sulfate, filter and concentrate under vacuum to give the crude product, which is purified by SFC to obtain the title compound.

[0460] Compound 246a: N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (P1, 10 mg, 0.015 mmol, yield: 27.3%), is a white solid.

[0461] 1 H NMR(400MHz,MeOD)δppm 7.76(t,J=8.2Hz,1H),7.46(s,1H),7.12–7.07(m,2H),6.83(s,1H),4.94(d,J=10.3Hz,1H),4.53– 4.67(m,3H),3.48–3.40(m,1H),3.37(t,J=5.1Hz,2H),3.23–3.13(m,2H),2.32–2.14(m,2H),2.09– 2.01(m,2H),1.90–1.81(m,2H),1.77–1.68(m,4H),1.61–1.48(m,1H),1.36–1.31(m,7H),1.27–1.1 4(m,2H),1.10(t,J=7.6Hz,3H),0.99–0.91(m,2H),0.88(d,J=6.3Hz,3H).MS(ESI)m / z=658.9[M+H] + .

[0462] Compound 246b: N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (P2, 10 mg, 0.015 mmol, yield: 27.3%), is a white solid.

[0463] 1 H NMR(400MHz,MeOD)δppm 7.73(t,J=8.1Hz,1H),7.46(s,1H),7.13–7.04(m,2H),6.83(s,1H),5.07(d,J=9.9Hz,1H),4.53–4.47(m ,3H),3.90–3.81(m,2H),3.61–3.56(m,1H),3.43–3.35(m,1H),3.25–3.19(m,1H),2.32–2.07(m,4H),2. 03–1.81(m,4H),1.79–1.71(m,3H),1.34(t,J=7.1Hz,4H),1.28–1.25(m,1H),1.21(d,J=6.9Hz,3H),1.1 9–1.08(m,1H),0.99–0.91(m,2H),0.88(d,J=6.3Hz,3H),0.83(t,J=7.6Hz,3H).MS(ESI)m / z=658.9[M+H] + .

[0464] Example 247:

[0465] Synthesis of N-[(2R,3S)-3-{4-[(2S)-2-[2-(3-cyanophenyl)-2,2-difluoroacetamyl]-2-[4-(difluoromethylene)cyclohexyl]acetamyl]-3-fluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]acrylamide

[0466]

[0467] Step 1: Stir a mixture of N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (100 mg, 0.29 mmol), (S)-{[(tert-butoxy)carbonyl]amino}[4-(difluoromethylene)cyclohexyl]acetic acid (87 mg, 0.29 mmol), HATU (163 mg, 0.43 mmol), and N,N-diisopropylethylamine (74 mg, 0.57 mmol) in THF (6 mL) at room temperature for 16 hours. The product was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (MeOH / DCM = 0-5%) to give ((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobutyl-2-yl)phenyl)amino)-2-oxoethyl)carbamate tert-butyl ester (90 mg, 0.14 mmol, yield: 48.7%), as a yellow solid. MS (ESI) m / z = 638.2 [M+H] + .

[0468] Step 2: Add TFA (1.5 mL) to a 2 mL solution of ((S)-1-(4-(difluoromethylene)cyclohexyl)-2-((2-fluoro-4-(((2S,3R)-4-(4-methylpiperazin-1-yl)-4-oxo-3-propamidobut-2-yl)phenyl)amino)-2-oxoethyl)carbamate (90 mg, 0.141 mmol) in DCM. Stir the mixture at room temperature for 2 hours. Then concentrate under vacuum to give crude N-((2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl)propionamide (110 mg). No further purification is required for the next step. MS(ESI)m / z = 538.2[M+H] + .

[0469] Step 3: A mixture of N-((2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl)propionamide (100 mg, 0.186 mmol), (3-cyanophenyl)difluoroacetic acid (37 mg, 0.186 mmol), HATU (106 mg, 0.279 mmol), and N,N-diisopropylethylamine (144 mg, 1.116 mmol) in THF (5 mL) was stirred at room temperature for 16 hours. The mixture was diluted with water (50 mL) and extracted with EtOAc (30 mL x 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (MeOH / DCM = 0-5%) to give N-[(2R,3S)-3-{4-[(2S)-2-[2-(3-cyanophenyl)-2,2-difluoroacetamido]-2-[4-(difluoromethylene)cyclohexyl]acetamido]-3-fluorophenyl}-1-(4-methylpiperazin-1-yl)-1-oxobut-2-yl]propionamide (12 mg, 0.017 mmol, yield: 9.0%), as a white solid. MS (ESI) m / z = 717.2 [M+H] + .

[0470] 1 H NMR (400MHz, CD3OD) δ8.00(s,1H),7.92(dd,J=13.1,7.9Hz,2H),7.68(dd,J=16.4,8.2Hz ,2H),7.19–7.02(m,2H),4.85(s,1H),4.41(d,J=8.7Hz,1H),3.68–3.41(m,3H),3.13(dt, J=13.7,6.8Hz,1H),2.77–2.59(m,2H),2.48(t,J=12.6Hz,2H),2.38(d,J=19.3Hz,3H),2 .23(td,J=15.1,7.4Hz,3H),2.10–1.69(m,7H),1.33(d,J=7.0Hz,3H),1.21–1.07(m,5H).

[0471] Example 248:

[0472] Synthesis of N-(1,1-dicyclopropyl-3-((4-(1-(4-(difluoromethylene)piperidin-1-yl)-1-oxopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0473]

[0474] Step 1: At 0°C, a solution of t-BuOK (7.13 g, 63.5 mmol) in THF (50 mL) was added to a solution of (methoxymethyl)triphenylphosphonium chloride (21.8 g, 63.5 mmol) in THF (150 mL). The mixture was stirred at room temperature for 1 hour under nitrogen atmosphere, followed by the addition of dicyclopropyl methyl ketone (5 g, 45.4 mmol) at this temperature, and further stirring was carried out at 25°C for 16 hours. The mixture was quenched with water (50 mL), diluted with hexane (600 mL), and then filtered. The filtrate was washed with water (100 mL x 3) and brine (100 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (EtOAc:hexane = 0–3%) to give a colorless oily product (6.1 g, 41.9 mmol, 92.3% yield). 1 HNMR(400MHz, CDCl3)δ5.87(s,1H),3.58(s,3H),1.85-1.78(m,1H),0.88–0.79(m ,1H),0.76–0.65(m,2H),0.66–0.56(m,2H),0.50–0.42(m,2H),0.27–0.18(m,2H).

[0475] Step 2: The mixture of (1-cyclopropyl-2-methoxyvinyl)cyclopropane (2 g, 14.5 mmol) in THF (40 mL) and 3NHCl (13 mL) was stirred at 55 °C for 4 hours and at room temperature for 16 hours. The mixture was diluted with hexane (200 mL), washed with water (50 mL * 2), dried with Na2SO4, and concentrated under vacuum to give a colorless oily crude product (1.7 g, crude product). 1 HNMR (400MHz, CDCl3) δ9.74 (d, J = 2.9Hz, 1H), 1.08-1.03 (m, 1H), 0.88–0.86 (m, 2H), 0.61–0.51 (m, 4H), 0.31–0.23 (m, 4H).

[0476] Step 3: A mixture of 2,2-dicyclopropylacetaldehyde (1 g, 8.1 mmol), TMSCN (1.21 g, 12.1 mmol), and ammonium carbonate (4.2 g, 43.7 mmol) in ethanol (8.5 mL) and water (8.5 mL) was stirred in a sealed tube at 95 °C for 6 hours. The mixture was acidified to pH 5 with HCl, diluted with DCM (100 mL), washed with water (30 mL x 3), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (MeOH:DCM = 0–5%) to give a white solid product (0.49 g, 2.0 mmol, 24.6% yield). MS (ESI) m / z = 195.2 [M+H] + .

[0477] Step 4: The mixture of 5-(dicyclopropylmethyl)imidazolidine-2,4-dione (490 mg, 2.52 mmol) in saturated NaOH (7N) (15 mL) was stirred at 105 °C for 72 hours. A solution of Boc₂O (1.1 g, 5 mmol) in THF (5 mL) was added at 25 °C. The mixture was further stirred at 25 °C for 2 hours. The mixture was diluted with EtOAc (80 mL) and washed with water (30 mL x 2). The combined aqueous solution was adjusted to pH 4 with HCl and then extracted with DCM (30 mL x 3). The combined organic layers were washed with water (20 mL), dried over Na₂SO₄, and concentrated to give a white solid product (330 mg, crude). MS (ESI) m / z = 292.1 [M + Na] + .

[0478] Step 5: A mixture of ethyl 2-(4-amino-3-fluorophenyl)propionate (400 mg, 1.89 mmol), 2-{[(tert-butoxy)carbonyl]amino}-3,3-dicyclopropylpropionic acid (510 mg, 1.89 mmol), and EDCI (726 mg, 3.79 mmol) in pyridine (10 mL) was stirred at 25 °C for 16 hours. The mixture was diluted with EtOAc (100 mL), washed with 1N HCl (20 mL x 3) and brine (20 mL), dried over Na₂SO₄, concentrated, and purified by rapid chromatography (EtOAc:hexane = 0–30%) to give the product (570 mg, 1.17 mmol, 61.8% yield) as a white solid. MS (ESI) m / z = 407.0 [M + H - 56] + . 1HNMR (400MHz, CDCl3) δ8.29–8.20(m,1H),8.12(s,1H),7.13–7.03(m,2H),5.52–5.33(m,1H),4.43-4.33(m,1H),4.19–4.05(m,2H),3.66(q,J= 7.1Hz,1H),1.54–1.35(m,12H),1.22(t,J=7.1Hz,3H),0.91(dd,J=10.5,6.6Hz,1H),0.76-0.68(m,2H),0.61–0.43(m,4H),0.32–0.20(m,4H).

[0479] Step 6: The mixture of ethyl 2-{4-[2-{[(tert-butoxy)carbonyl]amino}-3,3-dicyclopropylpropamido]-3-fluorophenyl}propionate (50 mg, 0.108 mmol) in a 2N HCl solution of MeOH (4 mL) was stirred at 25 °C for 4 hours. The mixture was diluted with DCM (30 mL), washed with saturated NaHCO3 (15 mL x 2) and water (15 mL), dried over Na2SO4, and concentrated to give a colorless oily product (40 mg, crude product). MS (ESI) m / z = 349.1 [M+H] + .

[0480] Step 7: A mixture of methyl 2-[4-(2-amino-3-dicyclopropylpropamido)-3-fluorophenyl]propionate (40 mg, 0.115 mmol), 2-isopropylpyrazole-3-carboxylic acid (18 mg, 0.115 mol), HATU (87 mg, 0.230 mmol), and DIEA (45 mg, 0.344 mmol) in THF (4 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by rapid chromatography (EtOAc:hexane = 0–30%) to give a yellow oily product (45 mg, 0.088 mmol, 76.8% yield). MS (ESI) m / z = 484.9 [M+H] + .

[0481] Step 8: Methyl 2-(4-{3,3-dicyclopropyl-2-[(2-isopropylpyrazol-3-yl)carbamoyl]propamidoyl}-3-fluorophenyl)propionate (45 mg, 0.093 mmol) and LiOH·H₂O (10 mg, 0.23 mmol) in THF / H₂O = 1:1 (3 mL) were stirred at 25 °C for 16 hours. The mixture was acidified to pH = 4 with 1 N HCl, diluted with DCM (50 mL), washed with water (20 mL) and brine (20 mL), dried over Na₂SO₄, and concentrated to give a yellow solid crude product (40 mg, crude product). MS (ESI) m / z = 471.1 [M+H] + .

[0482] Step 9: A mixture of 2-(4-{3,3-dicyclopropyl-2-[(2-isopropylpyrazol-3-yl)carbamoyl]propamidoyl}-3-fluorophenyl)propionic acid (40 mg, 0.085 mmol), 4-(difluoromethylene)piperidine (14 mg, 0.085 mmol), HATU (65 mg, 0.17 mmol), and DIEA (33 mg, 0.255 mmol) in THF (3 mL) was stirred at 25 °C for 16 hours. The mixture was diluted with EA (50 mL), washed with water (15 mL x 3) and brine (15 mL), dried over Na2SO4, concentrated, and purified by preparative TLC (MeOH:DCM = 3%) to give a white solid product (10.7 mg, 0.017 mmol, 20.4% yield). 1 HNMR(400MHz,CD3OD)δ7.92-7.88(m,1H),7.54(s,1H),7.16-7.11(m,2H),6.80(s, 1H),5.45-5.36(m,1H),4.98(d,J=7.1Hz,1H),4.19-4.13(m,1H),3.84–3.74(m,1H) ,3.58–3.41(m,3H),2.25–2.04(m,3H),1.69-1.65(m,1H),1.47(t,J=6.1Hz,6H),1. 41(d,J=6.8Hz,3H),0.95–0.79(m,3H),0.57–0.23(m,8H).MS(ESI)m / z=586.2[M+H] + .

[0483] Example 249:

[0484] Synthesis of N-((S)-1-((1r,4S)-4-(difluoromethyl)cyclohexyl)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0485]

[0486] Step 1: Palladium / carbon (40 mg) was added to a methanol (5 mL) solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetate (200 mg, 0.626 mmol), and the mixture was stirred overnight at room temperature under H2. The mixture was filtered through diatomaceous earth and concentrated under vacuum to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetate (200 mg, 0.622 mmol, yield: 99.4%) as a yellow oil. MS (ESI) m / z = 222.1 [M+H-Boc] + .

[0487] Step 2: LiOH·H₂O (65 mg, 1.56 mmol) was added to a THF / water solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid (200 mg, 0.62 mmol) in a THF / water ratio of 1:1 (6 mL). The mixture was stirred overnight at room temperature. It was diluted with water (20 mL) and extracted with ethyl acetate (20 mL). The pH of the aqueous phase was then adjusted to 3 with 1 N HCl and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid (160 mg, 0.52 mmol, yield: 83.6%) as a white solid. MS (ESI) m / z = 306.2 [MH] - .

[0488] Step 3: The mixture of N-((2R,3S)-3-(4-amino-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (40 mg, 0.10 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid (48 mg, 0.16 mmol), HATU (71 mg, 0.19 mmol), and DIEA (34 mg, 0.26 mmol) in THF (3 mL) was stirred at 25 °C for 48 hours. The product was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (elution gradient: ethyl acetate / petroleum ether = 0-50%) to give compound ((S)-1-((1r,4S)-4-(difluoromethyl)cyclohexyl)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)carbamate tert-butyl ester (70 mg, impure), as a pale yellow solid. MS (ESI) m / z = 673.3 [M+H] + .

[0489] Step 4: Add TFA (1 mL) to a solution of ((S)-1-((1r,4S)-4-(difluoromethyl)cyclohexyl)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)carbamate (70 mg, 0.104 mmol) in dichloromethane (3 mL) and stir the mixture at room temperature for 2 hours. Quench with saturated NaHCO3 (10 mL) and extract with dichloromethane (10 mL * 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product N-((2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-(difluoromethyl)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (50 mg, 0.087 mmol, yield: 83.6%), as a pale yellow solid. MS (ESI) m / z = 573.3 [M+H] + .

[0490] Step 5: The mixture of N-((2R,3S)-3-(4-((S)-2-amino-2-((1r,4S)-4-(difluoromethyl)cyclohexyl)acetamyl)-3-fluorophenyl)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxobut-2-yl)propionamide (50 mg, 0.087 mmol), 2-ethylpyrazole-3-carboxylic acid (18 mg, 0.131 mmol), HATU (60 mg, 0.157 mmol), and DIEA (28 mg, 0.218 mmol) in DMF (2 mL) was stirred at room temperature for 2 hours. It was then diluted with water (20 mL) and extracted with ethyl acetate (15 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by preparative HPLC (mobile phase: ACN-H₂O (0.1% FA), 35 / 65-75 / 25)) to give compound N-((S)-1-((1r,4S)-4-(difluoromethyl)cyclohexyl)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (8 mg, 0.012 mmol, yield: 13.2%) as a white solid. MS (ESI) m / z = 695.2 [M+H] + . 1 H NMR(400MHz,CD3OD)δ7.80–7.71(m,1H),7.46(d,J=2.0Hz,1H),7.16-7.05(m,2H) ,6.86-6.81(m,1H),5.95–5.45(m,1H),4.95(d,J=10.3Hz,1H),4.75(d,J=10.3Hz ,1H),4.57–4.44(m,2H),3.49-3.34(m,3H),3.25–3.11(m,2H),2.32–2.18(m,2H) ,2.17–1.95(m,3H),1.93–1.52(m,10H),1.38-1.25(m,7H),1.10(t,J=7.6Hz,3H).

[0491] Example 250:

[0492] Synthesis of N-((S)-2-((4-((2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1s,4R)-4-fluoro-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0493]

[0494] Step 1: MeLi (1.48 mL, 3.85 mmol, 2.6 M) was added to an 8 mL THF solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-oxocyclohexyl)acetate (500 mg, 1.75 mmol) at -78 °C and N2. The mixture was stirred at this temperature for 1 hour, followed by stirring at room temperature for 15 hours. The mixture was quenched with saturated NH4Cl (0.5 mL), diluted with DCM (30 mL), washed with water (10 mL) and brine (10 mL), dried over Na2SO4, concentrated, and purified by rapid chromatography (EtOAc:hexane = 0–50%) to give the product (160 mg, 0.32 mmol, yield: 1.82%) as a colorless oil. MS (ESI) m / z = 324.1 [M + Na] + .

[0495] Step 2: BAST (352 mg, 1.59 mmol) was added to a DCM (4 mL) solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-hydroxy-4-methylcyclohexyl)acetate (160 mg, 0.53 mmol) at 0 °C, and the mixture was stirred at 25 °C for 16 hours. The mixture was diluted with DCM (30 mL), washed with water (15 mL * 2), dried over Na₂SO₄, concentrated, and purified by rapid chromatography (EtOAc:hexane = 0–30%) to give the product (130 mg, 0.21 mmol, yield: 40.4%) as a colorless oil. MS (ESI) m / z = 326.1 [M + Na] + .

[0496] Step 3: A mixture of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-fluoro-4-methylcyclohexyl)acetate (130 mg, 0.43 mmol) and LiOH·H₂O (54 mg, 1.28 mmol) in THF / MeOH / H₂O (1 mL / 1 mL / 1 mL) was stirred at 25 °C for 3 hours. The mixture was concentrated and purified by preparative HPLC (Gemini-C18 150*21.2 mm, 5 μm: ACN---H₂O (0.1% TFA): 35-60) to give the product (45 mg, 0.078 mmol, yield: 18.2%) as a colorless oil. MS (ESI) m / z = 312.0 [M+Na] + .

[0497] Step 4: The mixture of (S)-{[(tert-butoxy)carbonyl]amino}(4-fluoro-4-methylcyclohexyl)acetic acid (40 mg, 0.14 mmol), N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (53 mg, 0.14 mmol), HATU (105 mg, 0.28 mmol), and DIEA (54 mg, 0.41 mmol) in THF (3 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by rapid chromatography (MeOH:DCM = 0–5%) to obtain the product, which was then purified by SFC (SP-120-10-C18-BIO-C18 250*50mm, 10µm (pH 8–10): CO2-MeOH(DEA)) to obtain the product (9 mg, 0.013 mmol, yield: 9.48%) as a colorless oil. MS (ESI) m / z = 655.1 [M+H] + .

[0498] Step 5: The mixture of N-[(S)-({4-[(2S,3R)-4-[4-(difluoromethylene)piperidin-1-yl]-4-oxo-3-propamidobutyl-2-yl]-2-fluorophenyl}carbamoyl)(4-fluoro-4-methylcyclohexyl)methyl]tert-butyl carbamate (9 mg, 0.014 mmol) in TFA / DCM (0.5 mL / 0.5 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily product (220 mg, crude product).

[0499] MS(ESI)m / z = 555.0 [M+H] + .

[0500] Step 6: Stir the mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-2-(4-fluoro-4-methylcyclohexyl)acetamyl]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (10 mg, 0.018 mmol), 2-ethylpyrazole-3-carboxylic acid (5 mg, 0.036 mmol), HATU (14 mg, 0.036 mol), and DIEA (19 mg, 0.14 mmol) in THF (2 mL) at room temperature for 16 hours. The mixture was diluted with DCM (40 mL), washed with water (15 mL x 3) and brine (15 mL), dried over Na₂SO₄, concentrated, and purified by preparative TLC (MeOH / DCM = 5%) to give the product N-((S)-2-((4-(2S,3R)-4-(4-(difluoromethylene)piperidin-1-yl)-4-oxo-3-propamidobutyl-2-yl)-2-fluorophenyl)amino)-1-((1S,4R)-4-fluoro-4-methylcyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (7.7 mg, 0.011 mmol, yield: 60%), as a white solid. MS (ESI) m / z = 677.1 [M+H] + . 1 H NMR (400MHz, CD3OD) δ7.78 (t, J=8.3Hz, 1H), 7.49 (d, J=2.0Hz, 1H), 7.17–7. 08(m,2H),6.89–6.85(m,1H),4.97(d,J=10.3Hz,1H),4.68–4.48(m,3H),3.5 1–3.37(m,3H),3.25-3.17(m,2H),2.31-2.17(m,4H),2.1-1.90(m,5H),1.8 6–1.72(m,2H),1.68–1.50(m,4H),1.39-1.33(m,9H),1.13(t,J=7.6Hz,3H).

[0501] Examples 251a and 251b:

[0502] N-((S)-2-((5-fluoro-1-((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and

[0503] Synthesis of N-((S)-2-((5-fluoro-1-((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0504]

[0505] Step 1: At -15°C, POCl3 (104 mg, 0.68 mmol) was added to a mixture of (2R)-2-(4-amino-5-fluoro-2-oxopyridin-1-yl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (100 mg, 0.34 mmol) and (S)-{[(tert-butoxy)carbonyl]amino}(4-methylcyclohexyl)acetic acid (92 mg, 0.34 mmol) in pyridine (5 mL). The mixture was stirred at this temperature for 4 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (EtOAc:hexane = 0-26%) to give a yellow oily product (35 mg, 0.048 mmol, 14.1% yield). MS (ESI) m / z = 549.0 [M+H] + .

[0506] Step 2: The mixture of N-[(S)-({5-fluoro-1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-2-oxopyridin-4-yl}carbamoyl)(4-methylcyclohexyl)methyl]tert-butyl carbamate (35 mg, 0.064 mmol) in TFA / DCM (0.5 mL / 0.5 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily product (40 mg, crude product). MS (ESI) m / z = 449.1 [M+H] + .

[0507] Step 3: A mixture of (2R)-2-{4-[(2S)-2-amino-2-(4-methylcyclohexyl)acetamyl]-5-fluoro-2-oxopyridin-1-yl}-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (40 mg, 0.089 mmol), 2-isopropylpyrazole-3-carboxylic acid (16 mg, 0.11 mmol), HATU (68 mg, 0.18 mmol), and DIEA (69 mg, 0.53 mmol) in THF (3 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and analyzed by preparative HPLC (Gemini-C18). 150*21.2mm, 5um: ACN-H2O (0.1% FA): 50-80) Purified to obtain products P1 and P2: N-((S)-2-((5-fluoro-1-((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-1-((1r,4S)-4-methylcyclohexyl)- 2-Oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and N-((S)-2-((5-fluoro-1-((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-1-((1r,4S)-4-methylcyclohexyl)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0508] Compound 251a: (35 mg, 0.0062 mmol, 6.95% yield), is a white solid.

[0509] 1 HNMR(400MHz,CD3OD)δ7.91-7.87(m,1H),7.77-7.73(m,1H),7.51(d,J=1.8Hz,1H) ,6.80(d,J=1.9Hz,1H),5.58(q,J=6.7Hz,1H),5.369-5.32(m,1H),4.59(d,J=8.2H z,1H),4.27–3.99(m,2H),3.33(s,3H),1.91-1.70(m,5H),1.51(d,J=6.8Hz,3H),1 .44(t,J=7.2Hz,6H),1.31–1.12(m,3H),1.02-0.89(m,2H),0.90(d,J=6.5Hz,3H).

[0510] MS(ESI)m / z = 585.0 [M+H] + .

[0511] Compound 251b: (6.3 mg, 0.011 mmol, 12.0% yield), is a white solid.

[0512] 1 HNMR(400MHz,CD3OD)δ7.93-7.88(m,1H),7.76-7.73(m,1H),7.51(d,J=1.9Hz,1H),6.7 9(d,J=2.0Hz,1H),5.59(q,J=6.7Hz,1H),5.41–5.30(m,1H),4.58(d,J=8.2Hz,1H),4.25 –4.01(m,2H),3.33(s,3H),1.92-1.70(m,5H),1.51(d,J=6.8Hz,3H),1.45(d,J=6.7Hz, 3H),1.43(d,J=6.8Hz,3H),1.31–1.12(m,3H),1.03-0.96(m,2H),0.90(d,J=6.5Hz,3H).

[0513] MS(ESI)m / z = 585.0 [M+H] + .

[0514] Examples 252a and 252b:

[0515] Synthesis of N-((S)-1-cycloheptyl-2-((5-fluoro-1-((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-2-oxo-1,2-dihydropyridin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazol-5-carboxamide

[0516]

[0517] Step 1: At -15°C, POCl3 (104 mg, 0.68 mmol) was added to a mixture of (2R)-2-(4-amino-5-fluoro-2-oxopyridin-1-yl)-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (100 mg, 0.34 mmol) and (S)-{[(tert-butoxy)carbonyl]amino}(cycloheptyl)acetic acid (92 mg, 0.34 mmol) in pyridine (3 mL). The mixture was stirred at -15°C for 4 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (EtOAc:hexane = 0-26%) to give a yellow oily product (35 mg, 0.051 mmol, 15.1% yield). MS (ESI) m / z = 549.0 [M+H] + .

[0518] Step 2: The mixture of N-[(S)-cycloheptayl({5-fluoro-1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-2-oxopyridin-4-yl}carbamoyl)methyl] tert-butyl carbamate (35 mg, 0.064 mmol) in TFA / DCM (0.5 mL / 0.5 mL) was stirred at 25 °C for 1 hour. The mixture was concentrated to give a yellow oily product (40 mg, crude product). MS (ESI) m / z = 449.0 [M+H] + .

[0519] Step 3: A mixture of (2R)-2-{4-[(2S)-2-amino-2-cycloheptylacetamyl]-5-fluoro-2-oxopyridin-1-yl}-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (40 mg, 0.089 mmol), 2-isopropylpyrazole-3-carboxylic acid (16 mg, 0.11 mmol), HATU (68 mg, 0.18 mmol), and DIEA (69 mg, 0.54 mmol) in THF (2 mL) was stirred at 25 °C for 16 hours. The mixture was concentrated and purified by preparative HPLC (Gemini-C18). 150*21.2mm, 5um: ACN-H2O (0.1% FA): 55-75) yielded the products N-((S)-1-cycloheptyl-2-((5-fluoro-1-((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and N-((S)-1-cycloheptyl-2-((5-fluoro-1-((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-2-oxo-1,2-dihydropyridin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide.

[0520] Compound 252a: P1 (6.6 mg, 0.011 mmol, 12.3% yield) is a white solid.

[0521] 1 HNMR(400MHz,CD3OD)δ7.94-7.89(m,1H),7.78-7.75(m,1H),7.53(d,J=1.9 Hz,1H),6.80(d,J=2.0Hz,1H),5.64–5.55(m,1H),5.41-5.32(m,1H),4.71( d,J=8.0Hz,1H),4.28–4.01(m,2H),3.35(s,3H),2.22-2.18(m,1H),1.89-1 .72(m,4H),1.69–1.55(m,4H),1.54–1.43(m,13H).MS(ESI)m / z=585.0[M+H] + .

[0522] Compound 252b: (2.7 mg, 0.0041 mmol, 7.36% yield), is a white solid.

[0523] 1 HNMR(400MHz,CD3OD)δ7.94-7.90(m,1H),7.77(d,J=5.3Hz,1H),7.53(d,J=1 .9Hz,1H),6.80(d,J=2.0Hz,1H),5.63-5.58(m,1H),5.40-5.33(m,1H),4.70 (d,J=8.0Hz,1H),4.24-4.05(m,2H),3.35(s,3H),2.24–2.18(m,1H),1.87-1 .72(m,4H),1.70–1.58(m,4H),1.55–1.43(m,13H).MS(ESI)m / z=585.0[M+H] + .

[0524] Examples 253a and 253b:

[0525] Synthesis of N-(1,1-dicyclopropyl-3-((4-(1-(4-(difluoromethylene)piperidin-1-yl)-1-oxopropyl-2-yl)-2-fluorophenyl)amino)-3-oxopropyl-2-yl)-1-isopropyl-1H-pyrazole-5-carboxamide

[0526]

[0527] Step 1: Add EDCI (181.50 mg, 0.947 mmol) to a solution of ethyl 2-(4-amino-3-fluorophenyl)propionate (100 mg, 0.473 mmol) and (S)-{[(tert-butoxy)carbonyl]amino}(cycloheptyl)acetic acid (128.46 mg, 0.473 mmol) in pyridine (10 mL). Stir the reaction mixture at 25 °C for 16 hours. The mixture was diluted with EtOAc (50 mL), washed with 1 N HCl (20 mL x 3) and brine (20 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (EtOAc / hexane = 0–30%) to give ethyl 2-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-cycloheptylacetamyl)-3-fluorophenyl)propionate (180 mg, 0.388 mmol, 81.9% yield). MS (ESI) m / z = 409.3 [M + H - 56] + .

[0528] Step 2: Add TFA (1 mL) to a DCM (4 mL) solution of ethyl 2-(4-((S)-2-((tert-Butoxycarbonyl)amino)-2-cycloheptaylacetamyl)-3-fluorophenyl)propionate (180 mg, 0.388 mmol). Stir the reaction mixture at 25 °C for 3 hours. Dilute the mixture with DCM (30 mL), wash with saturated NaHCO3 solution (15 mL x 2) and water (15 mL), dry with Na2SO4, and concentrate to obtain a colorless oily product, ethyl 2-(4-((S)-2-amino-2-cycloheptaylacetamyl)-3-fluorophenyl)propionate (160 mg, crude product). MS (ESI) m / z = 365.3 [M+H] + .

[0529] Step 3: HATU (333.84 mg, 0.878 mmol) and DIEA (170.22 mg, 1.317 mmol) were added to a THF (20 mL) solution of ethyl 2-(4-((S)-2-amino-2-cycloheptylacetamitano)-3-fluorophenyl)propionate (160 mg, 0.439 mmol) and 2-isopropylpyrazole-3-carboxylic acid (67.68 mg, 0.429 mmol). The reaction mixture was stirred at 25 °C for 16 hours. The mixture was concentrated under vacuum and purified by rapid chromatography (EtOAc / hexane = 0–20%) to give ethyl 2-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamitano)-3-fluorophenyl)propionate (180 mg, 0.360 mmol, 81.9% yield) as a pale yellow solid. MS(ESI)m / z = 501.4[M+H] + .

[0530] Step 4: LiOH·H2O (37.72 mg, 0.899 mmol) was added to a THF / H2O ratio of 1:1 (6 mL) of ethyl 2-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carbamoyl)acetamyl)-3-fluorophenyl)propionic acid (180 mg, 0.360 mmol). The reaction mixture was stirred at 25 °C for 16 hours. The mixture was adjusted to pH 4 with 1 N HCl, diluted with DCM (50 mL), washed with water (20 mL) and brine (20 mL), dried over Na2SO4 and concentrated under vacuum to give a yellow solid product, 2-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carbamoyl)acetamyl)-3-fluorophenyl)propionic acid (100 mg, crude product). MS(ESI)m / z = 473.3[M+H] + .

[0531] Step 5: Add HATU (160.91 mg, 0.423 mmol) and DIEA (82.05 mg, 0.635 mmol) to a THF (10 mL) solution of 2-(4-((S)-2-cycloheptyl-2-(1-isopropyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)propionic acid (100 mg, 0.212 mmol) and 4-(difluoromethylene)piperidine hydrochloride (35.89 mg, 0.212 mol). Stir the reaction mixture at 25 °C for 16 hours. The mixture was diluted with EtOAc (50 mL), washed with water (15 mL * 3) and brine (15 mL), dried with Na2SO4, concentrated under vacuum, and purified by rapid chromatography (MeOH / DCM = 0–3%) and chiral HPLC to obtain the products N-((S)-1-cycloheptyl-2-((4-(((S)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and N-((S)-1-cycloheptyl-2-((4-(((R)-1-(4-(difluoromethylene)piperidin-1-yl)-1-oxopropyl-2-yl)-2-fluorophenyl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide.

[0532] Compound 253a: (33.2 mg, 0.056 mmol, yield: 26.7%)

[0533] 1HNMR (400 MHz, DMSO-d6) δ 9.89 (s, 1H), 8.45 (d, J = 8.4 Hz, 1H), 7.71 (t, J = 8.3 Hz, 1H), 7.49 (d, J = 1.8 Hz, 1H), 7.18 (d, J = 11.9 Hz, 1H), 7.06 (d, J = 8.3 Hz, 1H), 6.91 (d, J = 1.9 Hz, 1H), 5.38 (dt, J = 13.3, 6.6 Hz, 1H), 4.58 (t, J = 8.3 Hz, 1H), 4.15 (q, J = 6.7 Hz, 1H), 3.62–3.38 (m, 4H), 2.02 (dt, J = 15.2, 12.0 Hz, 4H), 1.81–1.47 (m, 8H), 1.46–1.31 (m, 10H), 1.26 (t, J = 9.3 Hz, 4H). MS (ESI) m / z = 588.4 [M+H] + .

[0534] Compound 253b: (30.7 mg, 0.052 mmol, yield: 24.7%)

[0535] 1 HNMR (400 MHz, DMSO-d6) δ 9.90 (s, 1H), 8.46 (d, J = 8.5 Hz, 1H), 7.70 (t, J = 8.2 Hz, 1H), 7.49 (d, J = 1.9 Hz, 1H), 7.16 (dd, J = 11.8, 1.6 Hz, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.92 (d, J = 2.0 Hz, 1H), 5.38 (dt, J = 13.3, 6.6 Hz, 1H), 4.58 (t, J = 8.3 Hz, 1H), 4.15 (q, J = 6.8 Hz, 1H), 3.61–3.36 (m, 4H), 2.19–1.92 (m, 4H), 1.80–1.46 (m, 8H), 1.46–1.29 (m, 10H), 1.26 (t, J = 9.4 Hz, 4H). MS (ESI) m / z = 588.4 [M+H] + .

[0536] Example 254a and 254b:

[0537] Synthesis of N-((S)-1-cycloheptayl-2-((1-(((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and N-((S)-1-cycloheptayl-2-((1-(((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide)

[0538]

[0539] Step 1: Under nitrogen atmosphere, a mixture of 5-chloro-2H-pyridazin-3-one (1 g, 7.7 mmol), methyl (2S)-2-hydroxypropionate (1 g, 10 mmol), and PPh3 (3 g, 11.5 mmol) in THF (20 mL) was stirred at 0 °C for 10 min. Then, DIAD (2.34 g, 11.5 mmol / L) was added to the solution, and the mixture was stirred at 60 °C for 6 h under nitrogen atmosphere. The resulting mixture was diluted with water and extracted with EtOAc (20 mL x 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by rapid chromatography (EtOAc / hexane = 0-10%) to give a yellow solid product (2R)-2-(4-chloro-6-oxopyridazin-1-yl)propionate (1.4 g, 6.4 mmol, 83% yield).

[0540] MS(ESI)m / z=217.0[M+H]+.

[0541] Step 2: Under nitrogen atmosphere, a mixture of BocNH2 (1.5 g, 12.9 mmol), methyl (2R)-2-(4-bromo-5-fluoro-2-oxopyridin-1-yl)propionate (1.4 g, 6.46 mmol), Xantphos (308 mg, 0.65 mmol), Pd2(dba)3 (296 g, 0.3 mmol), and Cs2CO3 (6.3 g, 19 mmol) in dioxane (20 mL) was stirred at 110 °C for 5 hours. The resulting mixture was diluted with H2O (20 mL) and extracted with EtOAc (20 mL * 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (EtOAc / hexane = 0-50%) to give the yellow solid product (2R)-2-(4-{[(tert-butoxy)carbonyl]amino}-6-oxopyridazin-1-yl)propionate (650 mg, 2.1 mmol, 33.7% yield). MS (ESI) m / z = 298.1 [M+H] + .

[0542] Step 3: LiOH·H2O (275 mg, 6.56 mmol) was added to a MeOH / THF / H2O solution of (2R)-2-(4-{[(tert-butoxy)carbonyl]amino}-6-oxopyridazin-1-yl)propionate (650 mg, 2.19 mmol) in 3 mL of MeOH / THF / H2O = 1:1, and the mixture was stirred at 25 °C for 2 hours. The resulting mixture was diluted with saturated sodium carbonate solution (20 mL) and extracted with EtOAc (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product, which was purified by rapid chromatography (PE / EtOAc = 0-50%) to give the yellow solid product (2R)-2-(4-{[(tert-butoxy)carbonyl]amino}-6-oxopyridazin-1-yl)propionate (500 mg, 1.76 mmol, 80% yield).

[0543] MS(ESI)m / z = 284.0 [M+H] +

[0544] Step 4: A mixture of (2R)-2-(4-{[(tert-butoxy)carbonyl]amino}-6-oxopyridazin-1-yl)propionic acid (500 mg, 1.76 mmol), (2,2,2-trifluoroethyl)methylamine (316 mg, 2.12 mmol), and EDCI (676 mg, 3.53 mmol) in pyridine (10 mL) was stirred at 25 °C for 2 hours. The resulting mixture was diluted with 1N HCl (20 mL), extracted with EtOAc (20 mL * 3), and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (PE / EtOAc = 0-50%) to obtain N-{1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-6-oxopyridazin-4-yl} tert-butyl carbamate (400 mg, 1.0 mmol, 60% yield) as a yellow solid. MS(ESI)m / z = 379.1 [M+H] + .

[0545] Step 5: Dioxane hydrochloride (4M, 2mL) was added to a DCM (4mL) solution of N-{1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-6-oxopyridazin-4-yl}carbamate tert-butyl (350 mg, 0.92 mmol) and the resulting mixture was stirred at 25 °C for 16 hours. The mixture was concentrated under vacuum to give a yellow solid crude product (2R)-2-(4-amino-6-oxopyridazol-1-yl)-N-methyl-N-(2,2,2,2-trifluoroethyl)propionamide (140 mg, 0.50 mmol, 54.3% yield). MS (ESI) m / z = 279.0 [M+H] + .

[0546] Step 6: Add NaH (28 mg, 1.16 mmol) to a DMF (3 mL) solution of compound (2R)-2-(4-amino-6-oxopyridazol-1-yl)-N-methyl-N-(2,2,2,2-trifluoroethyl ester)propionamide (130 mg, 0.47 mmol) and stir at 25 °C for 1 hour. Add HATU (266 mg, 0.7 mmol) and DIEA (181 mg, 1.4 mmol) to another DMF (3 mL) solution of compound (S)-{[(tert-butoxy)carbonyl]amino}(cycloheptyl)acetic acid (353 mg, 0.93 mmol). Stir the mixture at 25 °C for 1 hour. Then add the mixture to the above reaction solution. Stir the resulting mixture at 25 °C for 16 hours. Quench the mixture with water (20 mL) and extract with EtOAc (40 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by chromatography (MeOH / DCM = 0-10%) to give a yellow solid, N-[(S)-cycloheptayl({1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-6-oxopyridazin-4-yl}carbamoyl)methyl]tert-butyl carbamate (40 mg, 0.08 mmol, 17% yield). MS (ESI) m / z = 532.2 [M+H] + .

[0547] Step 7: Add a solution of dioxane (4M, 2mL) of HCl to a DCM solution of N-[(S)-cycloheptayl({1-[(1R)-1-[methyl(2,2,2-trifluoroethyl)carbamoyl]ethyl]-6-oxopyridazin-4-yl}carbamoyl)methyl]carbamate tert-butyl ester (40 mg, 0.07 mmol) in 3 mL and stir at 25 °C for 3 hours. Concentrate the mixture under vacuum to obtain the crude product (2R)-2-{4-[(2S)-2-amino-2-cycloheptaylacetamitano]-6-oxypyridazin-1-yl}-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (40 mg), a yellow solid. MS (ESI) m / z = 432.1 [M+H] + .

[0548] Step 8: The mixture of compounds (2R)-2-{4-[(2S)-2-amino-2-cycloheptaylacetamyl]-6-oxypyridazin-1-yl}-N-methyl-N-(2,2,2-trifluoroethyl)propionamide (40 mg, 0.1 mmol), 2-isopropylpyrazole-3-carboxylic acid (21 mg, 0.14 mmol), HATU (88 mg, 0.24 mmol), and DIEA (72 mg, 0.56 mmol) in THF (3 mL) was stirred at 25 °C for 3 hours. The mixture was diluted with water (20 mL) and extracted with EtOAc (20 mL * 3). The organic layers were combined, dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (MeOH / DCM = 0-5%) and chiral HPLC to obtain N-((S)-1-cycloheptyl-2-((1-((R)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide and N-((S)-1-cycloheptyl-2-((1-((S)-1-(methyl(2,2,2-trifluoroethyl)amino)-1-oxopropyl-2-yl)-6-oxo-1,6-dihydropyridazin-4-yl)amino)-2-oxoethyl)-1-isopropyl-1H-pyrazole-5-carboxamide, both white solids.

[0549] Compound 254a: (2.9 mg, 0.056 mmol, yield: 26.7%)

[0550] 1 H NMR(400MHz, CD3ODδ8.05(d,J=2.5Hz,1H),7.50(d,J=2.1Hz,2H),6.78(d,J=1.9Hz,1H),5.75(q ,J=7.0Hz,1H),5.39–5.25(m,1H),4.51(d,J=8.0Hz,1H),4.32–4.15(m,1H),4.13–3.91(m,1H),3 .23(s,3H),2.21–2.08(m,1H),1.80(d,J=11.7Hz,1H),1.73(d,J=6.1Hz,3H),1.58(d,J=6.9Hz,4 H),1.53(d,J=10.0Hz,3H),1.43(dd,J=6.6,4.3Hz,9H),1.28(s,1H).MS(ESI)m / z=568.2[M+H]+.

[0551] Compound 254b: (3.6 mg, 0.052 mmol, yield: 24.7%)

[0552] 1 H NMR (400MHz, CD3OD) δ8.09(d,J=2.4Hz,1H),7.50(d,J=1.8Hz,1H),7.46(d,J=2.4Hz,1H),6.77(d,J=1.9Hz, 1H),5.75(t,J=6.9Hz,1H),5.42–5.27(m,1H),4.52(d,J=8.0Hz,1H),4.20(dt,J=18.6,7.7Hz,1H),4.06(dt, J=9.1,7.6Hz,1H),3.20(s,3H),2.15(dd,J=8.2,3.3Hz,1H),1.80(d,J=12.1Hz,1H),1.78–1.68(m,3H),1.5 7(d,J=6.9Hz,4H),1.55–1.48(m,3H),1.42(dt,J=10.9,5.5Hz,9H),1.28(s,1H).MS(ESI)m / z=568.2[M+H]+.

[0553] Example 255:

[0554] Synthesis of N-[(2R,3S)-3-{4-[(2S)-3,3-dicyclopropyl-2-[(2-ethylpyrazol-3-yl)carbamoyl]propamidoyl]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide

[0555]

[0556] Step 1: Stir a mixture of N-[(2R,3S)-3-(4-amino-3-fluorophenyl)-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (75 mg, 0.19 mmol), (2S)-2-{[(tert-butoxy)carbonyl]amino}-3,3-dicyclopropylpropionic acid (79 mg, 0.30 mmol), HATU (148 mg, 0.40 mmol), and N,N-diisopropylethylamine (101 mg, 0.78 mmol) in THF (5 mL) at room temperature for 16 hours. The product was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (MeOH / DCM = 0-5%) to give a yellow solid, N-[(1S)-2,2-dicyclopropyl-1-({4-[(2S,3R)-4-[4-(difluoromethylene)piperidin-1-yl]-4-oxo-3-propamidobutyl-2-yl]-2-fluorophenyl}carbamoyl)ethyl] tert-butyl carbamate (80 mg, 12.6 mmol, 66% yield). MS (ESI) m / z = 635.3 [M+H] + .

[0557] Step 2: Add TFA (1 mL) to a 2 mL solution of N-[(1S)-2,2-dicyclopropyl-1-({4-[(2S,3R)-4-[4-(difluoromethylene)piperidin-1-yl]-4-oxo-3-propamidobutyl-2-yl]-2-fluorophenyl}carbamoyl)ethyl]carbamate (60 mg, 0.09 mmol) in DCM. Stir the resulting mixture at room temperature for 3 hours. Then concentrate under vacuum to give the crude product N-[(2R,3S)-3-{4-[(2S)-2-amino-3,3-dicyclopropylpropamido]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobutyl-2-yl]propionamide (60 mg). No further purification is required for the next step. MS (ESI) m / z = 535.2 [M+H] + .

[0558] Step 3: A mixture of N-[(2R,3S)-3-{4-[(2S)-2-amino-3,3-dicyclopropylpropionamide]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (60 mg, 0.11 mmol), 2-ethylpyrazole-3-carboxylic acid (20 mg, 0.15 mmol), HATU (64 mg, 0.17 mmol), and N,N-diisopropylethylamine (58 mg, 0.45 mmol) in DMF (5 mL) was stirred at room temperature for 16 hours. The mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL x 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (MeOH / DCM = 0-5%) to give the white solid N-[(2R,3S)-3-{4-[(2S)-3,3-dicyclopropyl-2-[(2-ethylpyrazol-3-yl)carbamoyl]propamidoyl]-3-fluorophenyl}-1-[4-(difluoromethylene)piperidin-1-yl]-1-oxobut-2-yl]propionamide (12.6 mg, 0.018 mmol, 16.9% yield). MS (ESI) m / z = 657.3 [M+H] + . 1H NMR (400MHz, CD3OD) δ7.83(t,J=8.2Hz,1H),7.48(d,J=2.0Hz,1H),7.15(dd,J=12.0,1.4Hz,1H),7.05(d,J=8.3Hz,1H),6.83( d,J=2.0Hz,1H),4.94(dd,J=8.9,4.4Hz,2H),4.51(q,J=7.1Hz,2H),3.48(dd,J=11.9,5.6Hz,1H),3.44–3.31(m,2H),3.22–3.0 8(m,2H),2.33–2.17(m,2H),2.05(t,J=12.4Hz,2H),1.76–1.60(m,1H),1.55–1.41(m,1H),1.36(t,J=7.2Hz,3H),1.32(d,J=7 .0Hz,3H),1.10(t,J=7.6Hz,3H),0.83(ddd,J=14.9,9.2,4.8Hz,2H),0.77–0.70(m,1H),0.57–0.36(m,3H),0.34–0.13(m,5H).

[0559] Example 256:

[0560] Synthesis of 2-(1-((2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyryl)piperidin-4-yl)acetic acid

[0561]

[0562] Step 1: Add DIEA (55 mg, 0.425 mmol) to a DMF (5 mL) solution of (2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (50 mg, 0.081 mmol), methyl 2-(piperidin-4-yl)acetate hydrochloride (30 mg, 0.155 mmol), and HATU (37 mg, 0.097 mmol). Stir the reaction mixture at 25 °C for 4 hours. Pour the reaction mixture into water (20 mL) and extract with EtOAc (20 mL * 3). The organic layer was washed with brine (20 mL * 2), dried over anhydrous Na₂SO₄, and concentrated under vacuum to obtain the crude product. The crude product was purified by column chromatography (90-100% EtOAc / hexane) to give methyl 2-(1-((2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyryl)piperidin-4-yl)acetate (50 mg, 0.066 mmol, yield: 81.63%), as a yellow gel. MS (ESI) m / z = 758.3 [M+H] + .

[0563] Step 2: Add LiOH·H₂O (11.07 mg, 0.264 mmol) to a solution of methyl 2-(1-((2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyryl)piperidin-4-yl)acetate (50 mg, 0.066 mmol) in THF (10 mL) and water (5 mL). Stir the mixture at 25 °C for 3 hours. Adjust the pH of the mixture to 3 with 1N HCl, and then extract with EA (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by general-purpose (Genal) preparative HPLC (elution gradient: 18%-21% acetonitrile / water (0.1% FA)) to give the title compound 2-(1-((2R,3S)-2-(3-cyclopropyl-3-methylurea)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyryl)piperidin-4-yl)acetic acid (16.2 mg, 0.022 mmol, yield: 33.01%), as a white solid. MS (ESI) m / z = 744.3 [M+H] +. 1 H NMR(400MHz,MeOD)δppm 7.85–7.71(m,1H),7.51–7.46(m,1H),7.19–7.04(m,2H),6.87(d,J=2.0Hz,1H),4.98–4.88(m,1H),4.6 1(t,J=8.1Hz,1H),4.56–4.48(m,2H),4.38–4.26(m,1H),3.95–3.88(m,1H),3.19–3.09(m,1H),2.98–2. 82(m,4H),2.62–2.51(m,3H),2.48–2.35(m,1H),2.19–1.77(m,8H),1.65–1.48(m,2H),1.40–1.33(m,6 H),1.31–1.04(m,3H),0.95–0.88(m,2H),0.80–0.65(m,2H),0.60–0.49(m,0.5H),0.16–0.05(m,0.5H).

[0564] Examples 257a and 257b:

[0565] N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylureo)-4-((3R,4R)-4-methoxy-3,4-dimethylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide and

[0566] Synthesis of N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-((3R,4S)-4-methoxy-3,4-dimethylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0567]

[0568] Step 1: Under nitrogen atmosphere and at 0°C, methyl magnesium bromide (11.8 mL, 35.4 mmol) was added to a THF (100 mL) solution of 3-methyl-4-oxopiridine-1-carboxylic acid tert-butyl ester (5 g, 23.5 mol). The mixture was stirred at room temperature under N2 for 2 hours. The mixture was quenched with NH4Cl (30 mL) and extracted with EA (100 mL * 2). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under vacuum to give the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: 0-50% EA in PE) to give 4-hydroxy-3,4-dimethylpiperidine-1-carboxylic acid tert-butyl ester (2 g, 8.73 mmol, yield: 37.2%) as a yellow oil. MS (ESI) m / z = 156.0 [M-56-18] + .

[0569] Step 2: Under N2, tert-butyl 4-hydroxy-3,4-dimethylpiperidine-1-carboxylic acid (1 g, 4.34 mmol) was added to a solution of NaH (868 mg, 21.7 mmol) in DMF (10 mL). The mixture was stirred at room temperature under N2 for 1 hour. MeI (6.17 g, 43.4 mmol) and HMPA (0.5 mL) were added to the mixture, and the mixture was stirred at 50 °C for 12 hours. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, concentrated under vacuum, and the crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 5 / 1, v / v) to give tert-butyl 4-methoxy-3,4-dimethylpiperidine-1-carboxylic acid (500 mg, 2.06 mmol, yield: 47.1%) as a yellow oil. MS(ESI)m / z = 266.2[M+Na] + .

[0570] Step 3: Add hydrochloric acid solution (2 mL, 4 M dioxane solution) to a DCM (2 mL) solution of 100 mg (0.41 mmol) of 4-methoxy-3,4-dimethylpiperidine-1-carboxylic acid tert-butyl ester (DCM). Stir the mixture at room temperature for 2 hours. Concentrate the mixture under vacuum to give 4-methoxy-3,4-dimethylpiperidine hydrochloride (65 mg, 0.36 mmol, yield: 88.3%) as a yellow oil. This product can be used in the next step without further purification. MS (ESI) m / z = 144.2 [M+H] + .

[0571] Step 4: Add HATU (32 mg, 0.084 mmol) to a DMF (5 mL) solution of (2R,3S)-2-(3-cyclopropyl-3-methylureido)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (40 mg, 0.065 mmol), 4-methoxy-3,4-dimethylpiperidine hydrochloride (18 mg, 0.10 mmol), and DIEA (25 mg, 0.19 mmol). Stir the mixture at room temperature for 2 hours. The mixture was dissolved in EtOAc (50 mL), washed with H2O (20 mL) and brine (20 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. This crude product was purified by preparative HPLC (Gemini-C18 column, 5 μm silica, 21.2 mm diameter, 150 mm length) using a mixture of water (containing 0.05% FA) and MeCN as eluent (20-50%), with decreasing polarity. The purified product was then purified by SFC to give the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-((3R,4R)). -4-methoxy-3,4-dimethylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide and N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-methylurea)-4-((3R,4S)-4-methoxy-3,4-dimethylpiperidin-1-yl)-4-oxobut-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide.

[0572] Compound 257a (14.4 mg, 0.019 mmol, yield: 29.23%) is a white solid.

[0573] MS(ESI)m / z = 744.3[M+H] + .

[0574] 1H NMR (400MHz, CD3OD): δppm 7.96–7.80(m,1H),7.50(d,J=2.0Hz,1H),7.18–7.06(m,2H),6.88(d,J=2.4Hz,1H),6.29–6. 26(m,1H),4.95–4.91(m,1H),4.61–4.51(m,3H),4.15–3.34(m,3H),3.17–3.04(m,4H),2.95 –2.88(m,3H),2.61–2.44(m,4H),2.06–1.99(m,2H),1.87–1.79(m,4H),1.40–1.30(m,7H),1 .29–1.17(m,2H),1.09(s,1H),0.94–0.85(m,4H),0.78–0.66(m,5.5H),0.10–0.02(m,0.5H).

[0575] Compound 257b (18.9 mg, 0.025 mmol, yield: 38.46%) is a white solid.

[0576] MS(ESI)m / z = 744.3[M+H] + .

[0577] 1 H NMR (400MHz, CD3OD): δppm 7.91–7.79(m,1H),7.50–7.49(m,1H),7.17–7.05(m,2H),6.88–6.87(m,1H),6.29–6.25(m,1H),4.97– 4.90(m,1H),4.62–4.51(m,3H),4.14–3.87(m,1H),3.64–3.48(m,1H),3.22–2.99(m,5H),2.90–2.89( m,3H),2.61–2.45(m,4H),2.06–1.99(m,2H),1.94–1.87(m,3H),1.70–1.67(m,1H),1.40–1.33(m,7H) ,1.27–1.16(m,2H),1.11–1.04(m,1H),0.93–0.87(m,4H),0.82–0.74(m,5.5H),0.15–0.07(m,0.5H).

[0578] Example 258: Synthesis of N-((S)-2-((4-(2S,3R)-3-(3-cyclopropyl-3-ethylureo)-4-oxo-4-(4-(1-(trifluoromethyl)cyclopropyl)piperazin-1-yl)but-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide

[0579]

[0580] Step 1: At 0°C, add TEAA (491.82 mg, 2.34 mmol) to a DCM (20 mL) solution of methyl (2R,3S)-2-amino-3-(3-fluoro-4-nitrophenyl)butyrate (500 mg, 1.95 mmol) and TEA (592.39 mg, 5.85 mmol). Stir the mixture at room temperature for 2 hours. Dilute the mixture with water (40 mL) and extract with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-20%) to give methyl (2R,3S)-3-(3-fluoro-4-nitrophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (420 mg, 1.19 mmol, yield: 61.10%) as a yellow oil. MS (ESI) m / z = 352.8 [M+H] + .

[0581] Step 2: Add 10% Pd / C (80 mg) to a 10 mL solution of methyl (2R,3S)-3-(3-fluoro-4-nitrophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (420 mg, 1.19 mmol). Stir the mixture at room temperature under H2 atmosphere for 2 hours. Filter the mixture through diatomaceous earth and concentrate under vacuum to obtain a crude product, which is then purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-50%) to give methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (300 mg, 0.93 mmol, yield: 78.08%) as a pale yellow solid. MS (ESI) m / z = 323.1 [M+H] + .

[0582] Step 3: To a THF (5 mL) solution of methyl (2R,3S)-3-(4-amino-3-fluorophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (300 mg, 0.93 mmol), (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetic acid (426.41 mg, 1.39 mmol), DIEA (359.14 mg, 2.78 mmol), and HATU (528.96 mg, 1.39 mmol) were added. The reaction mixture was stirred at room temperature for 48 hours. The mixture was concentrated under vacuum to obtain a crude product, which was then purified by rapid chromatography (elution gradient: ethyl acetate / hexane = 0-40%) to give methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamityl)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamityl)butyrate (350 mg, 0.57 mmol, yield: 61.68%) as a white solid. MS (ESI) m / z = 632.1 [M+Na] + .

[0583] Step 4: Add TFA (3 mL) to a DCM (5 mL) solution of methyl (2R,3S)-3-(4-((S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamyl)butyrate (350 mg, 0.57 mmol). Stir the mixture at room temperature for 2 hours. Quench the mixture with saturated NaHCO3 (20 mL) and extract with dichloromethane (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: methanol / dichloromethane, 0–2%) to give methyl (2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamito)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamito)butyrate (280 mg, 0.55 mmol, yield: 95.72%), as a grayish-white solid. MS (ESI) m / z = 510.1 [M+H] + .

[0584] Step 5: To a DMF (20 mL) solution of methyl (2R,3S)-3-(4-((S)-2-amino-2-(4-(difluoromethylene)cyclohexyl)acetamyl)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamyl)butyrate (280 mg, 0.55 mmol), 1-ethyl-1H-pyrazole-5-carboxylic acid (92.24 mg, 0.66 mmol), DIEA (212.27 mg, 1.65 mmol), and HATU (229.27 mg, 0.60 mmol) were added. The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-50%) to give methyl (2R,3S)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (290 mg, 0.46 mmol, yield: 83.54%), as a grayish-white solid. MS (ESI) m / z = 632.2 [M+H] + .

[0585] Step 6: A solution of methyl (2R,3S)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)-2-(2,2,2-trifluoroacetamido)butyrate (280 mg, 0.44 mmol) in HCl / MeOH (5 mL) was stirred at 50 °C for 16 hours. The pH of the mixture was adjusted to 9 with a saturated NaHCO3 aqueous solution and extracted with DCM (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give methyl (2R,3S)-2-amino-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (150 mg, 0.28 mmol, yield: 63.18%), as a white solid. MS (ESI) m / z = 536.2 [M+H] + .

[0586] Step 7: Add triphosgene (53.20 mg, 0.18 mmol) and DIEA (144.54 mg, 1.12 mmol) to a DCM (10 mL) solution of methyl (2R, 3S)-2-amino-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (120 mg, 0.22 mmol). Stir the reaction mixture at room temperature for 10 minutes. Add N-ethylcyclopropylamine hydrochloride (54.50 mg, 0.45 mmol). Stir the resulting mixture at room temperature for 1 hour. Dilute the mixture with water (40 mL) and extract with DCM (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid chromatography (elution gradient: ethyl acetate / hexane, 0-50%) to give methyl (2R,3S)-2-(3-cyclopropyl-3-ethylureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (90 mg, 0.14 mmol, yield: 62.11%), as a yellow oil. MS (ESI) m / z = 647.2 [M+H] + .

[0587] Step 8: Add LiOH·H2O (29.20 mg, 0.70 mmol) to a MeOH / H2O (10 mL) solution of methyl (2R,3S)-2-(3-cyclopropyl-3-ethylureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyrate (90 mg, 0.14 mmol). Stir the reaction mixture at room temperature for 2 hours. Dilute the mixture with water (20 mL) and adjust the pH to 3 with 1N HCl, then extract with ethyl acetate (30 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give crude (2R,3S)-2-(3-cyclopropyl-3-ethylureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (60 mg, 0.095 mmol, yield: 68.14%), as a white solid. MS (ESI) m / z = 633.3 [M+H] + .

[0588] Step 9: Add 1-[1-(trifluoromethyl)cyclopropyl]piperazine (13.81 mg, 0.071 mmol), DIEA (18.34 mg, 0.14 mmol), and HATU (19.81 mg, 0.052 mmol) to a DMF (5 mL) solution of (2R,3S)-2-(3-cyclopropyl-3-ethylureo)-3-(4-((S)-2-(4-(difluoromethylene)cyclohexyl)-2-(1-ethyl-1H-pyrazole-5-carboxamido)acetamido)-3-fluorophenyl)butyric acid (30 mg, 0.047 mmol) in 1 mL of DMF. Stir the mixture at 25 °C for 2 hours. Dilute the mixture with water (10 mL) and extract with ethyl acetate (10 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. This crude product was purified by general-purpose (Genal) preparative HPLC (elution gradient: acetonitrile / water (0.1% FA)) to give the title compound N-((S)-2-((4-((2S,3R)-3-(3-cyclopropyl-3-ethylureo)-4-oxo-4-(4-(1-(trifluoromethyl)cyclopropyl)piperazin-1-yl)but-2-yl)-2-fluorophenyl)amino)-1-(4-(difluoromethylene)cyclohexyl)-2-oxoethyl)-1-ethyl-1H-pyrazole-5-carboxamide (21.2 mg, 0.026 mmol, yield: 55.27%), as a white solid. MS (ESI) m / z = 809.3 [M+H] + . 1 H NMR(400MHz,MeOD)δppm 7.81(t,J=8.2Hz,1H),7.39(d,J=2.0Hz,1H),7.10–6.93(m,2H),6.77(d,J=2.0Hz ,1H),6.16(d,J=9.2Hz,1H),4.81(t,J=9.6Hz,1H),4.54–4.40(m,3H),3.41–3.23 (m,3H),3.10–2.99(m,2H),2.67–2.26(m,6.5H),2.15–1.71(m,6.5H),1.31–1.09 (m,9H),1.00(t,J=7.2Hz,3H),0.87–0.80(m,4H),0.73(s,2H),0.67–0.60(m,2H).

[0589] Using a method similar to that used in the synthesis examples, the following compounds were obtained. The compounds are shown in Table 3.

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610] Synthesis of intermediates:

[0611] Synthesis of intermediate 1,2-((tert-butoxycarbonyl)amino)-3,3-dicyclopropylpropionic acid

[0612]

[0613] Step 1: At 0°C, a solution of t-BuOK (7.13 g, 63.5 mmol) in THF (50 mL) was added to a solution of (methoxymethyl)triphenylphosphonium chloride (21.8 g, 63.5 mmol) in THF (150 mL). The mixture was stirred at room temperature for 1 hour under nitrogen atmosphere, followed by the addition of dicyclopropyl methyl ketone (5 g, 45.4 mmol) at this temperature, and further stirring was carried out at 25°C for 16 hours. The mixture was quenched with water (50 mL), diluted with hexane (600 mL), and then filtered. The filtrate was washed with water (100 mL x 3) and brine (100 mL), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (EtOAc:hexane = 0–3%) to obtain the product (6.1 g, 41.9 mmol, 92.3% yield), a colorless oil. 1 HNMR(400MHz, CDCl3)δ5.87(s,1H),3.58(s,3H),1.85-1.78(m,1H),0.88–0.79(m ,1H),0.76–0.65(m,2H),0.66–0.56(m,2H),0.50–0.42(m,2H),0.27–0.18(m,2H).

[0614] Step 2: The mixture of (1-cyclopropyl-2-methoxyvinyl)cyclopropane (2 g, 14.5 mmol) in THF (40 mL) and 3NHCl (13 mL) was stirred at 55 °C for 4 hours, and then stirred at room temperature for 16 hours. The mixture was diluted with hexane (200 mL), washed with water (50 mL * 2), dried over Na2SO4, and concentrated under vacuum to give a colorless oily crude product (1.7 g, crude product). 1 HNMR (400MHz, CDCl3) δ9.74 (d, J = 2.9Hz, 1H), 1.08-1.03 (m, 1H), 0.88–0.86 (m, 2H), 0.61–0.51 (m, 4H), 0.31–0.23 (m, 4H).

[0615] Step 3: A mixture of 2,2-dicyclopropylacetaldehyde (1 g, 8.1 mmol), TMSCN (1.21 g, 12.1 mmol), and ammonium carbonate (4.2 g, 43.7 mmol) in ethanol (8.5 mL) and water (8.5 mL) was stirred in a sealed tube at 95 °C for 6 hours. The mixture was acidified to pH 5 with HCl, diluted with DCM (100 mL), washed with water (30 mL x 3), dried over Na₂SO₄, concentrated under vacuum, and purified by rapid chromatography (MeOH:DCM = 0–5%) to give a white solid product (0.49 g, 2.0 mmol, 24.6% yield). MS (ESI) m / z = 195.2 [M+H]+ .

[0616] Step 4: The mixture of 5-(dicyclopropylmethyl)imidazolidine-2,4-dione (490 mg, 2.52 mmol) in saturated NaOH (7N) (15 mL) was stirred at 105 °C for 72 hours. At 25 °C, 5 mL of THF containing 1.1 g (5 mmol) of Boc₂O was added. The mixture was stirred further at 25 °C for 2 hours. The mixture was diluted with EtOAc (80 mL) and washed with water (30 mL x 2). The combined water was acidified to pH 4 with HCl and then extracted with DCM (30 mL x 3). The combined organic layers were washed with water (20 mL), dried over Na₂SO₄, and concentrated to give a white solid product (330 mg, crude). MS (ESI) m / z = 292.1 [M + Na] + .

[0617] Synthesis of intermediate 2.(S)-2-((tert-Butoxycarbonyl)amino)-3,3-bicyclobutyric acid

[0618]

[0619] Step 1: At 0°C, magnesium bromo(cyclobutyl)methyl ether (42.8 mL, 21.4 mmol, 0.5 M THF solution) was added to a THF solution of cyclobutaneformaldehyde (1500 mg, 17.8 mmol) in 50 mL of THF, and the mixture was stirred at 0°C for 1.5 hours. The reaction mixture was quenched with saturated ammonium chloride (100 mL) and extracted with ethyl acetate (60 mL x 3). The organic layers were combined, washed with brine (60 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to give a colorless oily compound, dicyclobutylmethanol (750 mg, 5.4 mmol, yield: 30.0%). 1 H NMR (400MHz, CDCl3) δ3.39 (t, J = 6.9Hz, 1H), 2.36–2.23 (m, 2H), 1.95–1.72 (m, 12H).

[0620] Step 2: At 0°C, imidazole (425 mg, 6.2 mmol) and triphenylphosphine dibromide (2637 mg, 6.2 mmol) were added to a solution of dicyclobutylmethanol (730 mg, 5.2 mmol) in DCM (20 mL). The mixture was stirred overnight at room temperature. The reaction mixture was quenched with saturated ammonium chloride (50 mL) and extracted with DCM (50 mL x 3). The organic layers were combined, washed with brine (50 mL), dried over anhydrous sodium sulfate, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether) to give a colorless oily compound (bromomethylene)dicyclobutane (540 mg, 2.7 mmol, crude). No further purification was required for the next step.

[0621] Step 3: Under stirring, powdered sodium hydroxide (381 mg, 9.5 mmol) was added to a DMF (15 mL) solution of NI-(S)-BPB-GLY (950 mg, 1.9 mmol). Then, (bromomethylene)bicyclobutane (988 mg, 4.9 mmol) was added to the solution. The mixture was stirred under nitrogen, and the reaction progress was monitored by TLC. The reaction mixture was quenched with ice water (150 mL) and extracted with ethyl acetate (100 mL x 3). The organic layers were combined, washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, concentrated under vacuum to give a crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 1, v / v) to give the compound (280 mg, 0.45 mmol, yield: 10.1%) as a maroon solid. MS (ESI) m / z = 620.2 [M+H] + .

[0622] Step 4: The mixture of compound 6 (280 mg, 0.45 mmol) in 3M HCl / MeOH = 1:1 (15 mL) was stirred at 70 °C for 1 hour and concentrated. The residue was diluted with water (10 mL) and washed with DCM (10 mL * 2). Na₂CO₃ was added to the combined liquid layers to adjust the pH to 10, and THF (20 mL) and Boc₂O (492 mg, 2.26 mmol) were added. The mixture was stirred further at room temperature for 16 hours. The mixture was diluted with EA (50 mL) and washed with water (50 mL * 2). 4M HCl was added to the combined liquid layers to adjust the pH to 3. Then it was washed with EA (50 mL * 3) and the combined organic layers were concentrated to give the product (S)-2-((tert-Butoxycarbonyl)amino)-3,3-dicyclobutylpropionic acid (65 mg, 0.22 mmol, yield: 48.4%) as a yellow solid. MS(ESI)m / z = 320.2[M+Na] + .

[0623] Synthesis of intermediate 3,4-ethyl-1,2,5-oxadiazole-3-carboxylic acid

[0624]

[0625] Step 1: At 10°C, a solution of NaNO2 (7.95 g, 115 mmol) in H2O (25 mL) was added to a solution of methyl 3-oxovalerate (10 g, 76.8 mmol) in AcOH (60 mL), and the mixture was stirred at 25°C for 16 hours. The resulting mixture was quenched with saturated sodium bicarbonate solution (100 mL) and extracted with EtOAc (50 mL x 3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by rapid chromatography (EtOAc / hexane = 0-10%) to give a yellow oily product (2E)-2-(hydroxyimino)-3-oxovalerate methyl ester (8 g, 50 mmol, 65% yield). MS (ESI) m / z = 160.1 [M+H]+.

[0626] Step 2: A mixture of (2E)-2-(hydroxyimino)-3-oxovalerate methyl ester (8 g, 50 mmol), hydroxylamine hydrochloride (10.5 g, 150 mmol), and sodium acetate (8.2 g, 100 mmol) in ethanol (20 mL) was stirred at 80 °C for 16 hours. The resulting mixture was diluted with H₂O (100 mL) and extracted with EtOAc (100 mL * 3). The organic layers were combined, washed with brine (50 mL), dried over anhydrous sodium, filtered, and concentrated under vacuum to give a crude product, which was purified by rapid chromatography (EtOAc / hexane = 0-30%) to give a yellow oily product (2E,3E)-2,3-bis(hydroxyimino)valerate methyl ester (2.4 g, 13.7 mmol, 27.6% yield). 1 HNMR (400MHz, DMSO) δ12.10(s,1H),11.93(s,1H),3.73(s,3H),2.47(d,J=7.5Hz,1H),1.33–1.07(m,1H),1.01(t,J=7.5Hz,3H).

[0627] Step 3: At 25°C, a solution of (2E,3E)-2,3-bis(hydroxyimino)valerate (2.4 g, 13.8 mmol) and PIDA (5.3 g, 16.5 mmol) in DCM (30 mL) was stirred for 2 hours. The resulting mixture was concentrated under vacuum to give a crude product, which was purified by rapid chromatography (EtOAc / hexane = 0-10%) to give a yellow oily mixture of 4-ethyl-3-(methoxycarbonyl)-1,2,5-oxadiazole-2-onium-2-ester and 3-ethyl-4-(methoxycarbonyl)-1,2,5-oxadiazole 2-oxide (1.1 g, 6.39 mmol, 46% yield). 1 H NMR (400MHz, CDCl3) δ4.04(s,5H),3.98(s,3H),2.95(q,J=7.5Hz,2H),2.83(q,J=7.5Hz,4H),1.36(t,J=7.5Hz,3H),1.24(t,J=7.5Hz,6H).

[0628] Step 4: A solution of 10 mL of trimethyl phosphite in 4-ethyl-3-(methoxycarbonyl)-1,2,5-oxadiazole-2-onthium-2-ester (1.1 g, 6.4 mmol) was stirred at 100 °C for 16 hours. The resulting mixture was concentrated under vacuum to give a crude product, which was purified by rapid chromatography (EtOAc / hexane = 0-10%) to give a yellow oily mixture of methyl 4-ethyl-1,2,5-oxadiazole-3-carboxylic acid (200 mg, 1.28 mmol, 20% yield). 1 H NMR (400MHz, CDCl3) δ3.90 (s, 3H), 2.92 (q, J = 7.4Hz, 2H), 1.24 (t, J = 7.5Hz, 3H).

[0629] Step 5: LiOH·H2O (107 mg, 2.56 mmol) was added to a MeOH / THF / H2O solution of methyl 4-ethyl-1,2,5-oxadiazole-3-carboxylic acid (200 mg, 1.28 mmol) in 3 mL of MeOH / THF / H2O = 1:1:1, and the mixture was stirred at 25 °C for 2 hours. The resulting mixture was diluted with saturated sodium carbonate solution (10 mL) and extracted with DCM / MeOH = 10 / 1 (22 mL * 5). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a yellow oily crude product, 4-ethyl-1,2,5-oxadiazole-3-carboxylic acid (40 mg, 28 mmol, 22% yield). 1 H NMR (400MHz, CDCl3) δ3.03 (q, J = 7.5Hz, 2H), 1.37 (t, J = 7.5Hz, 3H).

[0630] Synthesis of intermediate 4.(S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid

[0631]

[0632] Step 1: Palladium / carbon (40 mg) was added to a methanol (5 mL) solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetate (200 mg, 0.626 mmol), and the mixture was stirred overnight at room temperature under H2. The mixture was filtered through diatomaceous earth and concentrated under vacuum to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetate (200 mg, 0.622 mmol, yield: 99.4%) as a yellow oil. MS (ESI) m / z = 222.1 [M+H-Boc] + .

[0633] Step 2: LiOH·H₂O (65 mg, 1.56 mmol) was added to a THF / water solution of (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid (200 mg, 0.62 mmol) in a THF / water ratio of 1:1 (6 mL). The mixture was stirred overnight at room temperature. It was diluted with water (20 mL) and extracted with ethyl acetate (20 mL). The pH of the aqueous phase was then adjusted to 3 with 1 N HCl and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (S)-2-((tert-butoxycarbonyl)amino)-2-(4-(difluoromethyl)cyclohexyl)acetic acid (160 mg, 0.52 mmol, yield: 83.6%) as a white solid. MS (ESI) m / z = 306.2 [MH] - .

[0634] Synthesis of intermediate 5.(S)-2-((tert-Butoxycarbonyl)amino)-2-((1S,4R)-4-fluoro-4-methylcyclohexyl)acetic acid

[0635]

[0636] Step 1: MeLi (1.48 mL, 3.85 mmol, 2.6 M) was added to an 8 mL THF solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-oxocyclohexyl)acetate (500 mg, 1.75 mmol) at -78 °C and N2. The mixture was stirred at this temperature for 1 hour, followed by stirring at room temperature for 15 hours. The mixture was quenched with saturated NH4Cl (0.5 mL), diluted with DCM (30 mL), washed with water (10 mL) and brine (10 mL), dried over Na2SO4, concentrated, and rapidly purified (EtOAc:hexane = 0–50%) to give the product (160 mg, 0.32 mmol, yield: 1.82%) as a colorless oil. MS (ESI) m / z = 324.1 [M + Na] + .

[0637] Step 2: At 0°C, BAST (352 mg, 1.59 mmol) was added to a DCM (4 mL) solution of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-hydroxy-4-methylcyclohexyl)acetate (160 mg, 0.53 mmol), and the mixture was stirred at 25°C for 16 hours. The mixture was diluted with DCM (30 mL), washed with water (15 mL * 2), dried over Na₂SO₄, concentrated, and purified by rapid chromatography (EtOAc:hexane = 0–30%) to give the product (130 mg, 0.21 mmol, yield: 40.4%) as a colorless oil. MS (ESI) m / z = 326.1 [M + Na] + .

[0638] Step 3: A mixture of (2S)-2-{[(tert-butoxy)carbonyl]amino}-2-(4-fluoro-4-methylcyclohexyl)acetate (130 mg, 0.43 mmol) and LiOH·H₂O (54 mg, 1.28 mmol) in THF / MeOH / H₂O (1 mL / 1 mL / 1 mL) was stirred at 25 °C for 3 hours. The mixture was concentrated and purified by preparative HPLC (Gemini-C18 150*21.2 mm, 5 μm: ACN---H₂O (0.1% TFA): 35-60) to give the product (45 mg, 0.078 mmol, yield: 18.2%) as a colorless oil. MS (ESI) m / z = 312.0 [M+Na] + .

[0639] Synthesis of intermediate 6,4-methyl-4,7-diazaspiro[2,5]octane

[0640]

[0641] Step 1: A mixture of 4,7-diazaspiro[2.5]octane-7-carboxylic acid tert-butyl ester (500 mg, 2.36 mmol), sodium cyanoborohydride (222 mg, 3.53 mmol), and formaldehyde aqueous solution (70.66 mg, 2.36 mol) in ACN (10 mL) was stirred at room temperature for 16 hours. The mixture was then concentrated under vacuum to obtain a crude product, which was purified by rapid chromatography (MeOH / DCM = 0-5%) to give a yellow solid product, 4-methyl-4,7-diazaspiro[2.5]octane-7-carboxylic acid tert-butyl ester (280 mg, 1.2 mmol, 52.4% yield). MS (ESI) m / z = 227.1 [M+H] + .

[0642] Step 2: A solution of dioxane (1.5 mL) in HCl was added to a DCM (2 mL) solution of 4-methyl-4,7-diazaspiro[2.5]octane-7-carboxylic acid tert-butyl ester (280 mg, 0.204 mmol). The resulting mixture was stirred at room temperature for 2 hours. The mixture was then concentrated under vacuum to obtain the crude product 4-methyl-4,7-diazaspiro[2.5]octane (230 mg, crude). No further purification was required for the next step. MS (ESI) m / z = 127.2 [M+H] + .

[0643] Synthesis of intermediate 7. (2R,3S)-1,2,3-trimethylpiperazine hydrochloride

[0644]

[0645] Step 1: Formaldehyde (83.67 mg, 2.79 mmol) was added to a DMF (10 mL) solution of (2S,3R)-2,3-dimethylpiperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.93 mmol). The reaction mixture was stirred at room temperature for 1 hour. Then, NaBH3CN (175.12 mg, 2.79 mmol) was added. The reaction mixture was stirred at room temperature for another 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (15 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give (2S,3R)-2,3,4-trimethylpiperazine-1-carboxylic acid tert-butyl ester (140 mg, 0.61 mmol, yield: 65.70%) as a pale yellow oil. MS (ESI) m / z = 229.0 [M+H] + .

[0646] Step 2: Add HCl dioxane (4M, 3mL, 12mmol) to a DCM solution of (2S,3R)-2,3,4-trimethylpiperazine-1-carboxylic acid tert-butyl ester (140 mg, 0.61 mmol) in 3 mL. Stir the reaction mixture at 25 °C for 2 hours. Concentrate the reaction mixture under vacuum to give a crude product. Dilute the mixture with Et₂O. Filter the mixture and dry the filter cake in air to give (2R,3S)-1,2,3-trimethylpiperazine hydrochloride (70 mg, 0.43 mmol, yield: 69.52%) as a white solid. MS (ESI) m / z = 129.2 [M+H] + .

[0647] Synthesis of intermediate 8.(S)-4-(difluoromethylene)-2-methylpiperidine hydrochloride

[0648]

[0649] Step 1: Under nitrogen protection and at -45°C, (S)-2-methyl-4-oxopiperidin-1-carboxylic acid tert-butyl ester (500 mg, 2.33 mmol) and 2-((difluoromethyl)sulfonyl)pyridine (540.89 mg, 2.80 mmol) were added to a DMF (50 mL) solution of t-BuOK (471.3 mg, 4.20 mmol). The reaction mixture was stirred at 25°C for 16 hours. 1 N HCl was added to the mixture to adjust the pH to 3, and the mixture was extracted with EA (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give (S)-4-(difluoromethylene)-2-methylpiperidine-1-carboxylic acid tert-butyl ester (350 mg, 1.42 mmol, yield: 60.37%) as a yellow oil. MS(ESI)m / z=191.9[M-56] + .

[0650] Step 2: HCl-dioxane (4M, 3mL, 12mmol) was added to a DCM solution of (S)-4-(difluoromethylene)-2-methylpiperidine-1-carboxylic acid tert-butyl ester (350 mg, 1.42 mmol) in 3 mL. The reaction mixture was stirred at 25 °C for 2 hours. The reaction was concentrated under vacuum to obtain a crude product, and the mixture was diluted with Et₂O. The mixture was filtered, and the filter cake was dried in air to give the product (S)-4-(difluoromethylene)-2-methylpiperidine HCl salt (220 mg, 1.20 mmol, yield: 84.86%) as a white solid. MS (ESI) m / z = 148.1 [M+H] + .

[0651] Synthesis of intermediate 9. (S)-1-(tert-butyl)-3-methylpiperazine

[0652]

[0653] Step 1: Under nitrogen atmosphere and stirring at 0°C, DIEA (1.94 g, 14.98 mmol) was added to a DCM (10 mL) solution of (S)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (1 g, 4.99 mmol) and acetic anhydride (765 mg, 7.49 mmol). The reaction mixture was stirred at 25°C for 16 hours. The solvent was removed under vacuum. The residue was diluted with water (10 mL). The mixture was extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product, which was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 3 / 1, v / v) to give (S)-4-acetyl-2-methylpiperazine-1-carboxylic acid tert-butyl ester (1.1 g, 4.54 mmol, yield: 90.92%) as a colorless gel. MS(ESI)m / z = 187.1 [M-56] + .

[0654] Step 2: Under nitrogen atmosphere at -10°C, titanium tetrachloride (4.13 mL, 4.13 mmol, 1 M DCM solution) was added to a stirred THF (10 mL) solution of (S)-4-acetyl-2-methylpiperazine-1-carboxylic acid tert-butyl ester (1 g, 4.13 mmol). The reaction mixture was stirred at -10°C for 30 min. At -10°C, methyl magnesium bromide solution (8.25 mL, 24.76 mmol, 1 M diethyl ether solution) was added to the mixture. The reaction mixture was stirred at 25°C for 16 h. The reaction mixture was quenched with 10% NaHCO3 solution (10 mL) at 0°C and filtered. The filtrate was concentrated under vacuum to obtain a residue. The residue was diluted with water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 20 / 1, v / v) to give (S)-4-(tert-butyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (400 mg, 1.56 mmol, yield: 37.80%) as a pale yellow gel. MS (ESI) m / z = 257.2 [M+H]+.

[0655] Step 3: Under nitrogen atmosphere, HCl (1.95 mL, 7.80 mmol, 4 M 1,4-dioxane solution) was added to a stirred solution of (S)-4-(tert-butyl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (400 mg, 1.56 mmol) in DCM (5 mL). The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under vacuum to give crude (S)-1-(tert-butyl)-3-methylpiperazine hydrochloride (120 mg, 0.62 mmol, yield: 39.91%), which was a yellow gel and could be used in the next step without further purification. 1 H NMR(400MHz,DMSO-d6)δ11.71(br.s,1H),9.93(br.s,1H),9.74(br.s,1H),3. 69–3.12(m,7H),1.38(s,9H),1.33(d,J=6.8Hz,3H).MS(ESI)m / z=157.3[M+H] + .

[0656] Synthesis of intermediate 10,1-(1-methylcyclopropyl)piperazine hydrochloride

[0657]

[0658] Step 1: At 0°C, Ac₂O (299.85 mg, 2.94 mmol) was added to a DCM (20 mL) solution of piperazine-1-carboxylic acid tert-butyl ester (500 mg, 2.67 mmol). The reaction mixture was stirred at 0°C for 1 hour. NaHCO₃ was added to the mixture to adjust the pH to 9, and the mixture was extracted with DCM (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give 4-acetylpiperazine-1-carboxylic acid tert-butyl ester (600 mg, 2.63 mmol, yield: 96.95%) as a pale yellow oil. MS (ESI) m / z = 173.1 [M-56] + .

[0659] Step 2: At -78°C, tetraisopropyl titanate (1487.47 mg, 5.23 mmol) was added to a THF (10 mL) solution of 4-acetylpiperazine-1-carboxylic acid tert-butyl ester (400 mg, 1.74 mmol). The reaction mixture was stirred at -78°C for 30 minutes. Ethyl magnesium bromide (3 M Et2O solution, 2.33 mL, 6.98 mmol) was added to the mixture at -78°C. The reaction mixture was stirred at -78°C for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (15 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-50% EtOAc / hexane) to give 4-(1-methylcyclopropyl)piperazine-1-carboxylic acid tert-butyl ester (200 mg, 0.83 mmol, yield: 47.50%) as a pale yellow oil. MS(ESI)m / z = 241.0[M+H] + .

[0660] Step 3: Add HCl-dioxane (4M, 3 mL, 12 mmol) to a DCM solution of 200 mg (0.83 mmol) of 4-(1-methylcyclopropyl)piperazine-1-carboxylic acid tert-butyl ester (3 mL). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the reaction mixture under vacuum to give a crude product. Dilute the mixture with Et₂O, filter, and dry the filter cake in air to give 1-(1-methylcyclopropyl)piperazine hydrochloride (100 mg, 0.57 mmol, yield: 68.19%) as a white solid. MS (ESI) m / z = 141.1 [M+H] + .

[0661] Synthesis of intermediate 11. (3S,5R)-4,4-difluoro-3,5-dimethylpiperidine hydrochloride

[0662]

[0663] Step 1: Under nitrogen atmosphere at 0°C, DAST (443 mg, 2.75 mmol) was added to a stirred solution of (3S,5R)-1-benzyl-3,5-dimethylpiperidin-4-one (300 mg, 1.37 mmol) in DCM (6 mL). The reaction mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into water (50 mL) and extracted with DCM (50 mL x 3). The organic layer was washed with brine (50 mL x 2), dried over anhydrous Na2SO4, and concentrated under vacuum to obtain the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 20% EtOAc in hexane solution) to give (3S,5R)-1-benzyl-4,4-difluoro-3,5-dimethylpiperidin (150 mg, 0.62 mmol, yield: 45.42%) as a colorless oil. MS(ESI)m / z = 240.0[M+H] + .

[0664] Step 2: Under nitrogen atmosphere, a solution of 1-chloroethyl chloroformate (173 mg, 1.62 mmol) in DCE (2 mL) was added to a stirred solution of (3S,5R)-1-benzyl-4,4-difluoro-3,5-dimethylpiperidine (150 mg, 0.62 mmol) in DCE (5 mL). The reaction mixture was stirred at 25 °C for 16 hours. The mixture was quenched with methanol (5 mL) and concentrated under vacuum to give the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 20% MeOH in DCM solution) to give (3S,5R)-4,4-difluoro-3,5-dimethylpiperidine hydrochloride (50 mg, 0.33 mmol, yield: 61.96%) as a yellow gel. MS (ESI) m / z = 150.0 [M+H] + .

[0665] Synthesis of intermediate 12,4-(difluoromethylene)-3-methylpiperidine hydrochloride

[0666]

[0667] Step 1: Under nitrogen protection and at -45°C, tert-butyl 3-methyl-4-oxopiperidin-1-carboxylic acid (1000 mg, 4.67 mmol) and 2-((difluoromethyl)sulfonyl)pyridine (1081.79 mg, 5.60 mmol) were added to a DMF (50 mL) solution of t-BuOK (942.59 mg, 8.40 mmol). The reaction mixture was stirred at 25°C for 16 hours. 1 N HCl was added to the mixture to adjust the pH to 3, and the mixture was extracted with EA (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give tert-butyl 4-(difluoromethylene)-3-methylpiperidine-1-carboxylic acid (800 mg, 3.24 mmol, yield: 68.99%) as a pale yellow oil. MS(ESI)m / z=192.2[M-56] + .

[0668] Step 2: Add HCl-dioxane (4M, 3 mL, 12 mmol) to a DCM solution of 4-(difluoromethylene)-3-methylpiperidine-1-carboxylic acid tert-butyl ester (800 mg, 3.22 mmol) in 3 mL. Stir the reaction mixture at 25 °C for 2 hours. Concentrate the reaction mixture under vacuum to give a crude product. Dilute the mixture with Et₂O, filter, and dry the filter cake in air to give 4-(difluoromethylene)-3-methylpiperidine hydrochloride (500 mg, 2.73 mmol, yield: 84.38%) as a white solid. MS (ESI) m / z = 148.2 [M+H] + .

[0669] Synthesis of intermediate 13,4-ethoxy-4-methylpiperidine hydrochloride

[0670]

[0671] Step 1: Under nitrogen atmosphere at 0°C, add NaH (140 mg, 3.5 mmol, 60% mineral oil solution) to a stirred DMF (5 mL) solution of 4-hydroxy-4-methylpiperidin-1-carboxylic acid tert-butyl ester (500 mg, 2.32 mmol). Stir the reaction mixture at 25°C for 1 hour. Add a DMF (5 mL) solution of EtI (725 mg, 4.65 mmol) to the above reaction mixture. Stir the reaction mixture at 25°C for another 16 hours. Quench the reaction with a saturated aqueous solution of NH4Cl (5 mL). Remove the solvent under vacuum. Dilute the residue with water (10 mL) and extract with ethyl acetate (10 mL x 3). The organic layers were combined, washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 4 / 1, v / v) to give 4-ethoxy-4-methylpiperidine-1-carboxylic acid tert-butyl ester (120 mg, 0.49 mmol, yield: 21.23%) as a colorless oil. MS (ESI) m / z = 266.2 [M + Na] + .

[0672] Step 2: Under nitrogen atmosphere, HCl (0.49 mL, 1.97 mmol, 4 M 1,4-dioxane solution) was added to a stirred solution of tert-butyl 4-ethoxy-4-methylpiperidine-1-carboxylic acid (120 mg, 0.49 mmol) in DCM (5 mL). The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated under vacuum to give crude 4-ethoxy-4-methylpiperidine hydrochloride (60 mg, 0.33 mmol, yield: 67.71%) as a colorless gel, which could be used in the next step without further purification. MS (ESI) m / z = 144.1 [M+H] + .

[0673] Synthesis of intermediate 14. (3aR, 6aS)-5-(difluoromethylene)octahydrocyclopentano[c]pyrrole hydrochloride

[0674]

[0675] Step 1: Under nitrogen protection, at -45°C, (3aR, 6aS)-5-oxohexahydrocyclopentane[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (2000 mg, 8.84 mmol) and 2-((difluoromethyl)sulfonyl)pyridine (2048.72 mg, 10.61 mmol) were added to a DMF (50 mL) solution of t-BuOK (1785.10 mg, 15.91 mmol). The reaction mixture was stirred at 25°C for 16 hours. 1N HCl was added to the mixture to adjust the pH to 3, and the mixture was extracted with EA (20 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give (3aR, 6aS)-5-(difluoromethylene)hexahydrocyclopentano[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (1200 mg, 4.63 mmol, yield: 51.72%) as a pale yellow oil. MS (ESI) m / z = 204.1 [M-56] + .

[0676] Step 2: HCl-dioxane (4 M, 3 mL, 12 mmol) was added to a DCM (3 mL) solution of (3aR, 6aS)-5-(difluoromethylene)hexahydrocyclopentano[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (1200 mg, 4.63 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction was concentrated under vacuum to give a crude product. The mixture was diluted with Et₂O, filtered, and the filter cake was dried in air to give (3aR, 6aS)-5-(difluoromethylene)octahydrocyclopentano[c]pyrrole hydrochloride (800 mg, 4.10 mmol, yield: 88.57%) as a white solid. MS (ESI) m / z = 160.1 [M+H] + .

[0677] Synthesis of intermediate 15. (3aR,6aS)-5,5-difluorooctahedrocyclopentano[c]pyrrole hydrochloride

[0678]

[0679] Step 1: At 0 °C, DAST (284.92 mg, 1.77 mmol) was added to a DCM (10 mL) solution of (3aR, 6aS)-5-oxohexahydrocyclopentano[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (200 mg, 0.88 mmol). The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (10 mL) and extracted with DCM (15 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give (3aR, 6aS)-5,5-difluorohexahydrocyclopentano[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (100 mg, 0.40 mmol, yield: 45.55%) as a pale yellow oil. MS(ESI)m / z=192.1[M-56] + .

[0680] Step 2: HCl-dioxane (4M, 3 mL, 12 mmol) was added to a DCM (3 mL) solution of (3aR, 6aS)-5,5-difluorohexahydrocyclopentano[c]pyrrole-2(1H)-carboxylic acid tert-butyl ester (100 mg, 0.40 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated under vacuum to give a crude product. The mixture was diluted with Et₂O, filtered, and the filter cake was dried in air to give (3aR, 6aS)-5,5-difluorooctahedralcyclopentano[c]pyrrole hydrochloride (40 mg, 0.22 mmol, yield: 54.00%) as a white solid.

[0681] Synthesis of intermediate 16,3,4-dimethylpiperidine-4-ol hydrochloride

[0682]

[0683] Step 1: At 0 °C, magnesium bromine (methyl)magnesium (3M Et₂O solution, 2.33 mL, 7.00 mmol) was added to a THF (30 mL) solution of 1000 mg, 4.67 mmol of 3-methyl-4-oxopiridine-1-carboxylic acid tert-butyl ester. The reaction mixture was stirred at room temperature for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (15 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-20% EtOAc / hexane) to give 4-hydroxy-3,4-dimethylpiperidine-1-carboxylic acid tert-butyl ester (800 mg, 3.49 mmol, yield: 74.40%) as a pale yellow oil. MS (ESI) m / z = 156.0 [M-18-56] + .

[0684] Step 2: HCl-dioxane (4M, 3 mL, 12 mmol) was added to a DCM solution of 100 mg (0.44 mmol) of tert-butyl 4-hydroxy-3,4-dimethylpiperidine-1-carboxylic acid (DCM) in 3 mL. The reaction mixture was stirred at 25 °C for 2 hours. The reaction was concentrated under vacuum to give a crude product. The mixture was diluted with Et₂O, filtered, and the filter cake was dried in air to give 3,4-dimethylpiperidine-4-ol hydrochloride (30 mg, 0.18 mmol, yield: 41.64%) as a white solid. MS (ESI) m / z = 130.2 [M+H] + .

[0685] Synthesis of intermediate 17,1-(piperazin-1-yl)cyclopropane-1-carboxynitrile

[0686]

[0687] Step 1: Add 1-aminocyclopropane-1-nitrile hydrochloride (200 mg, 1.69 mmol), N-benzyl-2-chloro-N-(2-chloroethyl)ethyl-1-amine hydrochloride (455 mg, 1.69 mol), and DIPEA (2 mL) to a sealed tube. Heat the mixture to 120 °C for 2 days. Cool the reaction to room temperature and concentrate under reduced pressure. Dilute the residue with EtOAc (30 mL), wash with water (20 mL x 2) and brine (20 mL). Dry the organic layer with sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography (0-60% EtOAc / hexane) to give 1-(4-benzylpiperazin-1-yl)cyclopropane-1-carboxynitrile (200 mg, 0.83 mmol, yield: 49.1%) as a yellow gel. MS (ESI) m / z = 242.1 [M+H] + .

[0688] Step 2: K₂CO₃ (344 mg, 2.49 mmol) was added to an acetone (10 mL) solution of 1-(4-benzylpiperazin-1-yl)cyclopropane-1-onitrile (200 mg, 0.83 mmol) and 1-chloroethylcarbonyl chloride (178 mg, 1.25 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The solvent was removed under vacuum. The residue was diluted with methanol (10 mL) and stirred at 60 °C for another 1 hour. The mixture was diluted with water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give crude 1-(piperazin-1-yl)cyclopropane-1-onitrile (50 mg, 0.33 mmol, yield: 39.9%) as a colorless gel, which could be used for the next step without further purification. MS (ESI) m / z = 152.3 [M+H] + .

[0689] Synthesis of intermediate 18,4-(dichloromethylene)piperidine hydrochloride

[0690]

[0691] Step 1: PPh3 (3.15 g, 0.012 mol) was added to a MeCN (20 mL) solution of 4-oxopiperidine-1-carboxylic acid tert-butyl ester (1 g, 0.005 mol). The mixture was stirred at 0 °C for 5 minutes under nitrogen. Then, bromotrichloromethane (1.2 g, 0.006 mol) was added at 0 °C. The mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with EtOAc (30 mL) and washed with water (20 mL x 2) and brine (20 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (0-30% EtOAc / hexane) to give 4-(dichloromethylene)piperidine-1-carboxylic acid tert-butyl ester (700 mg, 1.8 mmol, yield: 36.0%) as a colorless oil. MS (ESI) m / z = 263.2 [M+H] + .

[0692] Step 2: Add 1,4-dioxane / HCl (10 mL) to a DCM (10 mL) solution of 4-(dichloromethylene)piperidine-1-carboxylic acid tert-butyl ester (700 mg, 2.62 mmol). Stir the reaction at room temperature for 2 hours. Concentrate the mixture under reduced pressure to give 4-(dichloromethylene)piperidine hydrochloride (400 mg, 1.68 mmol, yield: 64.4%) as a yellow oil. 1 H NMR (400MHz, DMSO) δppm 7.64–7.55 (m, 1H), 3.08 (t, J = 5.6Hz, 4H), 2.68–2.65 (t, J = 6.4Hz, 4H).

[0693] Synthesis of intermediate 19,4-cyclohexenepropylpiperidine

[0694]

[0695] Step 1: Add t-BuOK (1.46 g, 0.013 mol) to a THF (30 mL) solution of bromo(cyclopropyl)triphenyl-15-phosphine (2.5 g, 0.006 mol). Stir the mixture at 50 °C for 1 hour under nitrogen. Cool the reaction to room temperature and add tert-butyl 4-oxopiperidin-1-carboxylic acid (1 g, 0.005 mol). Stir the reaction mixture at 60 °C for another 16 hours. Quench the mixture with water (50 mL) and extract with EtOAc (50 mL x 3). Wash the organic layer with brine (30 mL x 2), dry with anhydrous Na2SO4, and concentrate under vacuum to give the crude product. Purify the crude product by rapid silica gel chromatography (elution gradient: 0 to 10% hexane solution of EtOAc) to give tert-butyl 4-cyclopropylpiperidine-1-carboxylic acid (500 mg, 1.8 mmol, yield: 36.0%) as a white solid. 1 H NMR (400MHz, DMSO) δppm 3.35 (t, J = 5.6 Hz, 4H), 2.23 (t, J = 5.6 Hz, 4H), 1.41 (s, 9H), 1.02 (s, 4H).

[0696] Step 2: Add TFA (2 mL) to a DCM (10 mL) solution of 4-cyclomethoxypiperidine-1-carboxylic acid tert-butyl ester (500 mg, 2.23 mmol). Stir the reaction mixture at room temperature for 2 hours. Concentrate the mixture under reduced pressure, adjust the pH to 8 with NaHCO3, and extract with EtOAc (50 mL * 3). Wash the organic layer with brine (50 mL * 2), dry with anhydrous Na2SO4, and concentrate under vacuum to give the crude product, which is purified by rapid silica gel chromatography (elution gradient: 0 to 30% hexane solution of EtOAc) to give 4-cyclomethoxypiperidine (250 mg, 1.826 mmol, yield: 74.5%) as a white solid. MS (ESI) m / z = 124.2 [M + H] + .

[0697] Synthesis of intermediate 20,4-(tert-butoxy)piperidine

[0698]

[0699] Step 1: Under nitrogen atmosphere at 0°C, add boron trifluoride diethyl ether (880 mg, 6 mmol) to a stirred solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (2.5 g, 12.0 mmol) and tert-butyl 2,2,2-trichloroacetylimine (10.84 g, 49.0 mmol) in DCM (50 mL). Stir the reaction mixture at 25°C for 16 hours. Pour the reaction mixture into water (100 mL) and extract with DCM (100 mL x 3). Wash the organic layer with brine (100 mL x 2), dry with anhydrous Na2SO4 and concentrate under vacuum to give the crude product. Purify the crude product by rapid silica gel chromatography (elution gradient: 0 to 20% EtOAc in hexane) to give tert-butyl 4-(tert-butoxy)piperidine-1-carboxylate (350 mg, 1.2 mmol, yield: 9.68%) as a colorless oil. 1 HNMR (400MHz, DMSO) δ3.68–3.60(m,3H),2.95(s,2H),1.64–1.59(m,2H),1.38(s,9H),1.23–1.19(m,2H),1.13(s,9H).

[0700] Step 2: Under nitrogen atmosphere, add 0.5 mL of TFA to a stirred solution of 50 mg (0.19 mmol) of tert-butyl 4-(tert-butoxy)piperidine-1-carboxylic acid in 3 mL of DCM. Stir the reaction mixture at 25 °C for 3 hours. Concentrate the mixture under vacuum to give 4-(tert-butoxy)piperidine (25 mg, 0.16 mmol, yield: 82.17%) as a yellow gel. MS (ESI) m / z = 158.1 [M+H] + .

[0701] Synthesis of intermediate 21,2-(difluoromethylene)-6-azaspiro[3,4]octane 2,2,2-trifluoroacetate

[0702]

[0703] Step 1: At -40°C and under nitrogen, tert-butyl 2-oxo-6-azaspiro[3.4]octane-6-carboxylic acid (1 g, 4.42 mmol) and 2-(difluoromethane)sulfonylpyridine (1 g, 5.30 mmol) were added to a DMF (20 mL) solution of t-BuOK (893 mg, 7.95 mmol). The mixture was stirred at room temperature under N2 for 16 hours. The mixture was concentrated, the pH was adjusted to 1-2 with 1N hydrochloric acid aqueous solution, diluted with H2O (30 mL), and then extracted with ethyl acetate (30 mL * 3). The organic phases were combined, dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: 0-10% PE for EA) to give 2-(difluoromethylene)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (310 mg, 1.20 mmol, yield: 26.9%) as a yellow oil. MS (ESI) m / z = 204.0 [M-56+H] + .

[0704] Step 2: Add TFA (1 mL) to a DCM (5 mL) solution of 310 mg (1.19 mmol) of 2-(difluoromethylene)-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester (DCM). Stir the mixture at room temperature for 2 hours. Concentrate the mixture under vacuum to obtain a yellow, oily crude 2-(difluoromethylene)-6-azaspiro[3.4]octane TFA salt (320 mg). The crude product can be used for the next step without further purification. MS (ESI) m / z = 160.2 [M+H] + .

[0705] Synthesis of intermediate 22,6-(difluoromethylene)-2-azaspiro[3.3]heptane 2,2,2-trifluoroacetate

[0706]

[0707] Step 1: At -40°C and under nitrogen, tert-butyl 6-oxo-2-azaspiro[3.3]heptane-2-carboxylic acid (1 g, 4.73 mmol) and 2-(difluoromethane)sulfonylpyridine (1.1 g, 5.70 mmol) were added to a DMF (20 mL) solution of t-BuOK (956 mg, 8.52 mmol). The mixture was stirred at room temperature under N2 for 16 hours. The mixture was concentrated, the pH was adjusted to 1-2 with 1N hydrochloric acid aqueous solution, diluted with H2O (30 mL), and then extracted with ethyl acetate (30 mL * 3). The organic phases were combined, dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: EA solution of 0-10% PE) to obtain 6-(difluoromethylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (610 mg, 2.49 mmol, yield: 52.5%), which was a yellow oil. 1 H NMR (400MHz, CD3Cl3): δppm 3.94 (s, 4H), 2.84 (t, J = 3.8Hz, 4H), 1.43 (s, 9H). MS (ESI) m / z = 190.1 [M-56] + .

[0708] Step 2: Add TFA (1 mL) to a DCM (5 mL) solution of 6-(difluoromethylene)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (610 mg, 2.47 mmol). Stir the mixture at room temperature for 2 hours. Concentrate the mixture under vacuum to obtain a yellow, oily, crude 6-(difluoromethylene)-2-azaspiro[3.3]heptane TFA salt (400 mg). The crude product can be used in the next step without further purification. MS (ESI) m / z = 146.0 [M+H] + .

[0709] Synthesis of intermediate 23,4-methoxy-3,4-dimethylpiperidine hydrochloride

[0710]

[0711] Step 1: Under nitrogen atmosphere and at 0°C, methyl magnesium bromide (11.8 mL, 35.4 mmol) was added to a THF (100 mL) solution of 3-methyl-4-oxopiperidin-1-carboxylic acid tert-butyl ester (5 g, 23.5 mol). The mixture was stirred at room temperature under N2 for 2 hours. The mixture was quenched with NH4Cl (30 mL) and extracted with EtOAc (100 mL * 2). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under vacuum to give the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: 0-50% EtOAc in PE solution) to give 4-hydroxy-3,4-dimethylpiperidin-1-carboxylic acid tert-butyl ester (2 g, 8.73 mmol, yield: 37.2%) as a yellow oil. MS (ESI) m / z = 156.0 [M-56-18] + .

[0712] Step 2: Under N2, tert-butyl 4-hydroxy-3,4-dimethylpiperidine-1-carboxylic acid (1 g, 4.34 mmol) was added to a solution of NaH (868 mg, 21.7 mmol) in DMF (10 mL). The mixture was stirred under N2 at room temperature for 1 hour. MeI (6.17 g, 43.4 mmol) and HMPA (0.5 mL) were added to the mixture, and the mixture was stirred at 50 °C for 12 hours. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate, concentrated under vacuum, and the crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 5 / 1, v / v) to give tert-butyl 4-methoxy-3,4-dimethylpiperidine-1-carboxylic acid (500 mg, 2.06 mmol, yield: 47.1%) as a yellow oil. MS(ESI)m / z = 266.2[M+Na] + .

[0713] Step 3: Add hydrochloric acid solution (2 mL, 4 M dioxane solution) to a DCM solution of 100 mg (0.41 mmol) of tert-butyl 4-methoxy-3,4-dimethylpiperidine-1-carboxylic acid (tert-butyl ester) in 2 mL of DCM. Stir the mixture at room temperature for 2 hours. Concentrate the mixture under vacuum to give 4-methoxy-3,4-dimethylpiperidine HCl salt (65 mg, 0.36 mmol, yield: 88.3%) as a yellow oil. The product can be used in the next step without further purification.

[0714] MS(ESI)m / z = 144.2[M+H] + .

[0715] Synthesis of intermediate 24. (3S,5R)-4-methoxy-3,4,5-trimethylpiperidine hydrochloride

[0716]

[0717] Step 1: Under nitrogen protection, methyl magnesium bromide (2.3 mL, 6.9 mmol) was added to a THF (10 mL) solution of (3S,5R)-1-benzyl-3,5-dimethylpiperidin-4-one (300 mg, 1.38 mol) at 0 °C. The mixture was stirred at room temperature under N2 for 2 hours. The mixture was quenched with NH4Cl (10 mL) and extracted with EA (20 mL * 2). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: 0-50% EA in PE solution) to give (3S,5R)-1-benzyl-3,4,5-trimethylpiperidin-4-ol (300 mg, 1.29 mmol, yield: 93.2%) as a yellow oil. MS (ESI) m / z = 234.0 [M+H] + .

[0718] Step 2: Add 10% Pd / C (100 mg) to a MeOH (10 mL) solution of (3S,5R)-1-benzyl-3,4,5-trimethylpiperidin-4-ol (300 mg, 1.29 mmol). Stir the mixture at room temperature under H2 for 16 hours. Filter the mixture and concentrate under reduced pressure to give (3S,5R)-3,4,5-trimethylpiperidin-4-ol (170 mg, 1.19 mmol, yield: 92.4%) as a yellow oil. MS (ESI) m / z = 144.1 [M+H] + .

[0719] Step 3: Triethylamine (318 mg, 3.14 mmol) and (Boc)₂O (274 mg, 1.26 mmol) were added to a DCM (10 mL) solution of (3S,5R)-3,4,5-trimethylpiperidin-4-ol (150 mg, 1.05 mmol). The mixture was stirred at room temperature for 2 hours. The mixture was concentrated under vacuum to give a residue. The residue was subjected to rapid chromatography (silica gel, solvent gradient: 0-30% PE in EtOAc solution) to give (3S,5R)-4-hydroxy-3,4,5-trimethylpiperidin-1-carboxylic acid tert-butyl ester (200 mg, 0.82 mmol, yield: 78.4%) as a yellow oil. MS (ESI) m / z = 170.2 [M-56-18] + .

[0720] Step 4: Under nitrogen protection, (3S,5R)-4-hydroxy-3,4,5-trimethylpiperidine-1-carboxylic acid tert-butyl ester (200 mg, 0.82 mmol) was added to a solution of NaH (164 mg, 4.09 mmol) in DMF (10 mL). The mixture was stirred at room temperature under N2 for 1 hour. MeI (1.16 g, 8.19 mmol) and HMPA (0.2 mL) were added to the mixture, and the mixture was stirred at 50 °C for 12 hours. The mixture was quenched with water (20 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic layers were dried over anhydrous sodium sulfate and concentrated under vacuum to obtain a crude product. This crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 5 / 1, v / v) to give (3S,5R)-4-methoxy-3,4,5-trimethylpiperidine-1-carboxylic acid tert-butyl ester (160 mg, 0.62 mmol, yield: 75.8%) as a yellow oil. MS (ESI) m / z = 280.2 [M + Na] + .

[0721] Step 5: Add hydrochloric acid solution (2 mL, 4 M dioxane solution) to a DCM (2 mL) solution of (3S,5R)-4-methoxy-3,4,5-trimethylpiperidine-1-carboxylic acid tert-butyl ester (160 mg, 0.62 mmol). Stir the mixture at room temperature for 2 hours. Concentrate the mixture under vacuum to give (3S,5R)-4-methoxy-3,4,5-trimethylpiperidine hydrochloride (110 mg, 0.57 mmol, yield: 91.7%) as a yellow oil. The product can be used in the next step without further purification. MS (ESI) m / z = 158.2 [M+H] + .

[0722] Synthesis of intermediate 25,4-isopropyl-4-methoxypiperidine

[0723]

[0724] Step 1: Under nitrogen atmosphere at -40°C, a solution of tetrahydrofuran containing magnesium bromo(isopropyl)magnesium (37.5 mL, 37.5 mmol) was added dropwise to a stirred 50 mL THF solution of tert-butyl 4-oxopiridine-1-yl)carboxylate (5 g, 25 mmol). The reaction mixture was stirred at -40°C for 4 hours. The residue was quenched with 1N HCl (50 mL) and then extracted with EtOAc (200 mL x 3). The organic layer was washed with brine (150 mL x 2), dried over anhydrous Na2SO4, and concentrated under vacuum to give the crude product. The crude product was purified by rapid silica gel chromatography (elution gradient: 0 to 30% EtOAc in hexane) to give tert-butyl 4-hydroxy-4-isopropylpiperidine-1-carboxylate (600 mg, 2.5 mmol, yield: 10.00%) as a colorless oil. MS(ESI)m / z = 170.0 [M-18-56] + .

[0725] Step 2: At 0°C, NaH (163.70 mg, 4.09 mmol) was added to a DMF (10 mL) solution of 4-hydroxy-4-isopropylpiperidine-1-carboxylic acid tert-butyl ester (200 mg, 0.82 mmol). The reaction mixture was stirred at room temperature for 1 hour. At 0°C, MeI (1162.27 mg, 8.19 mmol) and HMPA (1 mL) were added. The reaction mixture was stirred at 50°C for 16 hours. The mixture was quenched with water (10 mL) and extracted with EtOAc (15 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give 4-isopropyl-4-methoxypiperidine-1-carboxylic acid tert-butyl ester (180 mg, 0.70 mmol, yield: 85.09%) as a pale yellow oil. MS(ESI)m / z = 280.1[M+Na] + .

[0726] Step 3: Add HCl-dioxane (3 mL, 12 mmol) to a DCM solution of 180 mg (0.70 mmol) of 4-isopropyl-4-methoxypiperidine-1-carboxylic acid tert-butyl ester (DCM). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the reaction mixture under vacuum to give 4-isopropyl-4-methoxypiperidine HCl salt (100 mg, 0.64 mmol, yield: 90.93%) as a white solid. MS (ESI) m / z = 158.1 [M+H] + .

[0727] Synthesis of intermediate 26,2-methyl-1-(1-(trifluoromethyl)cyclopropane)piperazine

[0728]

[0729] Step 1: A mixture of 2-(benzylamino)ethane-1-ol (5 g, 0.03 mol) and 2-methylethylene oxide (1584.2 mg, 5.86 mmol) was stirred at 50 °C for 16 hours. The mixture was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under vacuum to give a crude product. This crude product was purified by rapid silica gel chromatography (elution gradient: petroleum ether / ethyl acetate, 50 / 1, v / v) to give 1-(benzyl(2-hydroxyethyl)amino)propane-2-ol (5.5 g, 0.023 mol, yield: 71.65%) as a yellow oil. MS (ESI) m / z = 210.0 [M+H] + .

[0730] Step 2: At 0°C, SOCl2 (5657.4 mg, 47.5 mmol) was added to a stirred solution of 1-(benzyl(2-hydroxyethyl)amino)prop-2-ol (2000 mg, 9.51 mmol) in 30 mL of DCE. The reaction mixture was stirred at 70°C for 16 hours. The mixture was concentrated under vacuum to give a crude product. The crude product was diluted with water (100 mL) and extracted with dichloromethane (100 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under vacuum to give N-benzyl-2-chloro-N-(2-chloroethyl)prop-1-amine hydrochloride (2200 mg, 6.98 mmol, yield: 73.40%) as a colorless oil. MS (ESI) m / z = 245.9 [M + H] + .

[0731] Step 3: N-benzyl-2-chloro-N-(2-chloroethyl)propane-1-amine hydrochloride (500 mg, 2.02 mmol), 1-(trifluoromethyl)cyclopropylamine (326.8 mg, 2.02 mol), and DIEA (4 mL) were placed in a sealed tube. The mixture was heated to 120 °C for 2 days. The reaction was cooled to room temperature and concentrated under reduced pressure. The residue was diluted with EtOAc (30 mL) and washed with water (20 mL x 2) and brine (20 mL). The organic layer was dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (0-60% EtOAc / hexane) to give 4-benzyl-2-methyl-1-(1-(trifluoromethyl)cyclopropyl)piperazine (100 mg, 0.53 mmol, yield: 14.9%) as a yellow gel. MS (ESI) m / z = 298.9 [M+H]+.

[0732] Step 4: 4-Benzyl-2-methyl-1-(1-(trifluoromethyl)cyclopropyl)piperazine (100 mg, 0.334 mmol) and 10% Pd / C (22 mg, 0.1 mmol) were suspended in methanol (8 mL). The reaction mixture was stirred at 50 psi for 6 hours at room temperature. The mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to give 2-methyl-1-(1-(trifluoromethyl)cyclopropyl)piperazine (40 mg, 0.17 mmol, yield: 51.7%) as a colorless gel. MS (ESI) m / z = 209.1 [M+H]+

[0733] Synthesis of intermediate 27,4-ethyl-4-(methoxy-d3)piperidine

[0734]

[0735] Step 1: At 0°C, NaH (698.17 mg, 17.45 mmol) was added to a DMF (10 mL) solution of 4-ethyl-4-hydroxypiperidine-1-carboxylic acid tert-butyl ester (800 mg, 3.49 mmol). The reaction mixture was stirred at room temperature for 1 hour. Iodomethane-d3 (4.01 g, 34.91 mmol) was added to the mixture at 0°C. The reaction mixture was stirred at 50°C for 16 hours. The mixture was quenched with water (30 mL) and extracted with EA (15 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give 4-ethyl-4-(methoxy-d3)piperidine-1-carboxylic acid tert-butyl ester (700 mg, 2.84 mmol, yield: 81.45%) as a pale yellow oil. MS(ESI)m / z = 269.0 [M + Na] + .

[0736] Step 2: Add HCl-dioxane (3 mL, 12 mmol) to a DCM solution of 100 mg (0.40 mmol) of 4-ethyl-4-(methoxy-d3)piperidine-1-carboxylic acid tert-butyl ester (DCM). Stir the reaction mixture at 25 °C for 2 hours. Concentrate the reaction mixture under vacuum to give 4-ethyl-4-(methoxy-d3)piperidine HCl salt (40 mg, 0.27 mmol, yield: 67.93%) as a white solid. MS (ESI) m / z = 147.2 [M+H] + .

[0737] Synthesis of intermediate 28.1-(1-(difluoromethyl)cyclopropane)piperazine

[0738]

[0739] Step 1: Add 1-(difluoromethyl)cyclopropane-1-amine hydrochloride (200 mg, 1.39 mmol), N-benzyl-2-chloro-N-(2-chloroethyl)ethyl-1-amine hydrochloride (375 mg, 1.39 mmol), and DIPEA (2 mL) to a sealed tube. Heat the mixture to 120 °C for 2 days. Cool the reaction to room temperature and concentrate under reduced pressure. Dilute the residue with EtOAc (30 mL) and wash with water (20 mL x 2) and brine (20 mL). Dry the organic layer with sodium sulfate, filter, and concentrate under reduced pressure. Purify the residue by column chromatography (0-60% EtOAc / hexane) to give 1-benzyl-4-(1-(difluoromethyl)-cyclopropyl)piperazine (300 mg, 1.12 mmol, yield: 80.55%) as a yellow gel. MS (ESI) m / z = 267.2 [M+H] + .

[0740] Step 2: 1-Benzyl-4-(1-(difluoromethyl)cyclopropyl)piperazine (300 mg, 1.12 mmol) and 10% Pd / C (477 mg, 4.48 mmol) were suspended in MeOH (10 mL). The reaction mixture was stirred at room temperature under 50 psi hydrogen for 6 hours. The mixture was filtered through a diatomaceous earth pad, and the filtrate was concentrated under reduced pressure to give 1-(1-(difluoromethyl)cyclopropyl)piperazine (80 mg, 0.45 mmol, yield: 40.46%) as a white solid. MS (ESI) m / z = 177.2 [M+H] + .

[0741] Synthesis of intermediate 29.1-(piperazin-1-yl)cyclopropane-1-carboxylic acid methyl ester

[0742]

[0743] Step 1: N-benzyl-2-chloro-N-(2-chloroethyl)ethyl-1-amine hydrochloride (3.58 g, 0.0096 mol) was added to a DIEA (30 mL) solution of methyl 1-aminocyclopropane-1-carboxylate (1 g, 0.0087 mol). The reaction mixture was stirred in a sealed tube at 120 °C for 48 hours. The mixture was diluted with water (10 mL) and extracted with EA (15 mL x 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The residue was purified by column chromatography (0-10% EtOAc / hexane) to give methyl 1-(4-benzylpiperazin-1-yl)cyclopropane-1-carboxylate (400 mg, 1.46 mmol, yield: 16.79%) as a colorless oil. MS (ESI) m / z = 275.1 [M+H] + .

[0744] Step 2: Add 10% Pd / C (50 mg) to a MeOH (10 mL) solution of methyl 1-(4-benzylpiperazin-1-yl)cyclopropane-1-carboxylate (150 mg, 0.54 mmol). Stir the reaction mixture at 25 °C for 2 hours. Filter the mixture and concentrate under vacuum to give methyl 1-(piperazin-1-yl)cyclopropane-1-carboxylate (70 mg, 0.38 mmol, yield: 69.49%) as a pale yellow oil. MS (ESI) m / z = 185.2 [M+H] + .

[0745] Synthesis of intermediate 30. (1-(piperazin-1-yl)cyclopropane)methanol

[0746]

[0747] Step 1: LiAlH4 (1.09 mL, 1.09 mmol) was added to a THF (10 mL) solution of methyl 1-(4-benzylpiperazin-1-yl)cyclopropane-1-carboxylate (150 mg, 0.54 mmol). The reaction mixture was stirred at 25 °C for 2 hours. The mixture was diluted with water (10 mL) and extracted with EA (15 mL * 3). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give (1-(4-benzylpiperazin-1-yl)cyclopropyl)methanol (100 mg, 0.41 mmol, yield: 74.25%) as a yellow oil. MS (ESI) m / z = 247.1 [M + H] + .

[0748] Step 2: Add 10% Pd / C (30 mg) to a solution of (1-(4-benzylpiperazin-1-yl)cyclopropyl)methanol (100 mg, 0.41 mmol) in MeOH (10 mL). Stir the reaction mixture at 25 °C for 2 hours. Filter the mixture and concentrate under vacuum to give (1-(piperazin-1-yl)cyclopropyl)methanol (60 mg, 0.38 mmol, yield: 94.61%) as a pale yellow oil. MS (ESI) m / z = 157.1 [M+H] + .

[0749] Synthesis of intermediate 31,4-ethoxy-4-(trifluoromethyl)piperidine

[0750]

[0751] Step 1: Over 15 minutes, trimethyl(trifluoromethyl)silane (1.8 g, 12.86 mol) was added to a THF (30 mL) solution of 4-oxopiperidin-1-carboxylic acid benzyl ester (2 g, 8.57 mmol) cooled in an ice bath via a feeding funnel. A 1.0 MTBAF THF solution (85.7 mL, 85.7 mmol) was slowly added dropwise to the mixture. The mixture was stirred at room temperature for 16 hours. The mixture was concentrated to a volume of 20 mL, carefully quenched with 1 N hydrochloric acid aqueous solution, diluted with H₂O (30 mL), and then extracted with DCM (50 mL x 3). The organic phases were combined, dried over Na₂SO₄, filtered, and concentrated under vacuum to obtain the crude product. The crude product was subjected to rapid chromatography (silica gel, solvent gradient: 0-30% PE in EA) to give 4-hydroxy-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (1.6 g, 5.28 mmol, yield: 61.5%) as a white solid. MS (ESI) m / z = 304.1 [M+H] + .

[0752] Step 2: Under N2, add 4-hydroxy-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (500 mg, 1.65 mmol) to a solution of NaH (330 mg, 8.24 mmol) in 10 mL of DMF. Stir the mixture under N2 at room temperature for 1 hour. Add EtI (2.57 g, 16.5 mmol) to the mixture and stir at 50 °C for 12 hours. Quench the mixture with water (20 mL) and extract with ethyl acetate (50 mL x 2). Dry the combined organic layers with anhydrous sodium sulfate, concentrate under vacuum to obtain the crude product, which is purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 5 / 1, v / v) to give 4-ethoxy-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (480 mg, 1.45 mmol, yield: 87.9%) as a yellow oil. MS (ESI) m / z = 332.2 [M+H] + .

[0753] Step 3: Add 10% pd / C (100 mg) to a MeOH (10 mL) solution of 4-ethoxy-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (480 mg, 1.45 mmol). Stir the mixture at room temperature under H2 for 16 hours. Filter the mixture and concentrate under vacuum to give 4-ethoxy-4-(trifluoromethyl)piperidine (180 mg, 0.91 mmol, yield: 63.0%) as a yellow oil.

[0754] Synthesis of intermediate 32,4-(trifluoromethyl)piperidine-4-ol

[0755]

[0756] Step 1: Add 10% pd / C (100 mg) to a MeOH (10 mL) solution of 500 mg (1.65 mmol) of benzyl 4-hydroxy-4-(trifluoromethyl)piperidine-1-carboxylate. Stir the mixture at room temperature under H2 for 16 hours. Filter the mixture and concentrate under vacuum to give 4-(trifluoromethyl)piperidine-4-ol (160 mg, 0.95 mmol, yield: 57.3%) as a yellow oil. MS (ESI) m / z = 170.1 [M+H] + .

[0757] Synthesis of intermediate 33,2-(difluoromethylene)-7-azaspiro[3,5]nonane hydrochloride

[0758]

[0759] Step 1: Under nitrogen atmosphere at -40°C, a solution of t-BuOK (697 mg, 6.21 mmol) in DMF (4 mL) was added to a solution of 2-oxo-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (745 mg, 3.11 mmol) and DFMPS (600 mg, 3.11 mmol). The mixture was stirred at 25°C for 2 hours. The mixture was neutralized with 1N HCl, diluted with hexane (80 mL), washed with water (30 mL x 2) and brine (30 mL x 2), dried over Na2SO4, concentrated, and purified by rapid chromatography (EtOAc: hexane = 0–10%) to give a colorless oily product 2-(difluoromethylene)-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (609 mg, 2.23 mmol, 71.9% yield). 1 H NMR (400MHz, CDCl3) δ3.40-3.25 (m, 4H), 2.45-2.35 (m, 4H), 1.57 (dd, J = 7.4, 3.6Hz, 4H), 1.45 (s, 9H).

[0760] Step 2: A solution of 2-(difluoromethylene)-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (609 mg, 2.23 mmol) in 1,4-dioxane / HCl (3 mL) and DCM (3 mL) was stirred at 25 °C for 2 hours. The mixture was concentrated to give a white solid crude product, 2-(difluoromethylene)-7-azaspiro[3.5]nonane hydrochloride (422 mg, 2.02 mmol, 90.6% yield). MS (ESI) m / z = 174.2 [M+H]+.

[0761] Synthesis of intermediate 34,4-(2,2-difluoroethoxy)-4-(trifluoromethyl)piperidine

[0762]

[0763] Step 1: Under N2, add 4-hydroxy-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (300 mg, 0.99 mmol) to a solution of NaH (118.7 mg, 2.97 mmol) in DMF (10 mL). Stir the mixture at room temperature for 1 hour. Add 2,2-difluoroethyltrifluoromethanesulfonate (635.4 mg, 2.97 mmol) to the mixture and stir at 50 °C for 16 hours. Quench the mixture with water (20 mL) and extract with ethyl acetate (50 mL x 2). Dry the combined organic layers with anhydrous sodium sulfate, concentrate under vacuum to give a crude product, which is purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 5 / 1, v / v) to give 4-(2,2-difluoroethoxy)-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (310 mg, 0.84 mmol, yield: 85.3%) as a yellow oil. MS(ESI)m / z = 368.1 [M+H] + .

[0764] Step 2: Add 10% Pd / C (50 mg) to a methanol (10 mL) solution of 4-(2,2-difluoroethoxy)-4-(trifluoromethyl)piperidine-1-carboxylic acid benzyl ester (310 mg, 0.84 mmol). Stir the mixture at room temperature under H2 for 4 hours. Filter the mixture and concentrate under vacuum to give 4-(2,2-difluoroethoxy)-4-(trifluoromethyl)piperidine (120 mg, 0.51 mmol, yield: 61.0%) as a yellow oil. MS (ESI) m / z = 234.1 [M+H] + .

[0765] Synthesis of intermediate 35.N-methyl-4-(trifluoromethyl)piperidine-4-amine

[0766]

[0767] Step 1: Under nitrogen atmosphere, formaldehyde (112 mg, 1.49 mol, 40% aqueous solution) and acetic acid (0.2 mL) were added to a stirred solution of 4-amino-4-(trifluoromethyl)piperidine-1-carboxylic acid tert-butyl ester (400 mg, 1.49 mmol) in MeOH (5 mL). The reaction mixture was stirred at 25 °C for 30 min. A suspension of NaBH3CN (140 mg, 2.23 mmol) in MeOH (5 mL) was added to the reaction mixture. The reaction mixture was stirred at 25 °C for another 2 h. The reaction was quenched with saturated NH4Cl aqueous solution (5 mL). The solvent was removed under vacuum. The residue was diluted with water (10 mL). The mixture was extracted with ethyl acetate (10 mL x 3). The organic layers were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give the crude product. The crude product was purified by rapid chromatography (elution gradient: petroleum ether / ethyl acetate, 1 / 5, v / v) to give tert-butyl 4-(methylamino)-4-(trifluoromethyl)piperidine-1-carboxylic acid (80 mg, 0.283 mmol, yield: 19.01%) as a colorless gel. MS (ESI) m / z = 227.2 [M-56] + .

[0768] Step 2: Under nitrogen atmosphere, HCl (0.28 mL, 1.13 mmol, 4 M 1,4-dioxane solution) was...

Claims

1. A compound of formula (I) or its enantiomers, stereoisomers, solvates or pharmaceutically acceptable salts thereof: Its features are, Excerpt A is selected from the following group: A 5-12 membered heteroaryl group containing one or more nitrogen atoms, wherein the heteroaryl group is optionally substituted by one or more substituents selected independently from the group consisting of: halogen, hydroxyl, cyano, -NH2, C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 cycloalkyl, C 3-7 Cycloalkoxy, phenyl, phenoxy, 5-6-membered heteroaryl and 4-7-membered heterocyclic alkyl, wherein the 5-6-membered heteroaryl and 4-7-membered heterocyclic alkyl contain one or more heteroatoms each independently selected from O, S and N; -NH-C(O)-R3, where R3 is selected from the following group: C 3-12 Cycloalkyl and 3-12 membered heterocyclic groups, C 5-12 Aryl and 5-12 heteroaryl groups, which are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, halogen, -CN, C 1-6 Cyanoalkyl, oxo, -NH2, and -OH; the heterocyclic alkyl and heteroaryl groups contain one or more heteroatoms, each independently selected from O, S, and N; Optionally substituted methyl groups, wherein one or more substituents are independently selected from: C 1-6 Alkoxy, C 6-12 aryl groups and 5-12 membered heteroaryl groups containing one or more heteroatoms independently selected from O, S, and N, each optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy groups, halogens, -CN, and -OH; -OC 3-7 cycloalkyl and -O-CH2-C 3-7 Cycloalkyl, -O-CH2-5-7-membered heteroaryl, and benzyl, each benzyl, C 3-7 The cycloalkyl and heteroaryl groups are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 cycloalkyl, halogen, -CN, -NH2 and -OH; -NH(C 1-6 alkyl), -NH(C) 3-7 cycloalkyl), -N(C) 1-6 alkyl)2 and -N(C 1-6 Alkyl)(C 3-7 cycloalkyl groups, each optionally substituted by one or more substituents selected independently from the group consisting of: C 1-6 Alkyl, C 1-6 hydroxyalkyl, C 1-6 Alkoxy, C 3-7 cycloalkyl, halogen, -CN, -NH2 and -OH; C 1-6 Alkyl, C 6-12 Aryl, 5-12 membered heteroaryl, 4-7 membered heterocyclic, each optionally substituted by one or more substituents independently selected from the group consisting of: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocyclic alkyl, halogen, halogenated C 1-6 Alkyl, -CN, -NH2, -OH, phenyl, -NH(C 1-6 Alkyl group), -(CH2) q -NH(CO)-(C 1-4 Alkyl), -NHS(O)2(C 1-6 Alkyl); q is 0, 1, 2, 3 or 4; Alternatively, two substituents located on the same or adjacent ring atoms and the ring atom attached to them together form: substituted or unsubstituted C 5-7 Carbocyclic group, or substituted or unsubstituted 5-7 membered heterocyclic group; Part B is selected from C 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more heteroatoms independently selected from N, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens; the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups; part C is selected from the group consisting of: * A 4-9 membered heterocyclic group containing one or more heteroatoms selected from O, N, and S, wherein the heterocyclic group is optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene, C... 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-8 carbonyl group, -OC 3-6 cycloalkyl, C 1-6 Hydroxyalkyl, halogen, -CN, CF3, -CD3, -NH2, =NH, -OH, -COOH, C 1-6 Alkoxy, -SC 1-6 Alkyl, C 1-3 carbonyl, oxo, -N(C) 1-6 alkyl)2、-NH(C 1-6 alkyl), -NHC(O)-(C 1-6 alkyl), -NHC(O)-NH(C) 1-6 Alkyl), -NHS(O)2-(C 1-6 Alkyl), -NHS(O)2-(C 3-6 cycloalkyl), -S(O)2-(C 1-6 Alkyl), -S(O)2-(C 3-6 cycloalkyl), -C(O)-C 1-6 Alkyl, 4-7 membered heterocyclic and 5-7 membered heteroaryl; The substituent is optionally substituted by one or more substituents, each independently selected from the group consisting of: deuterium, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkyl groups, halogens, -CN, CF3, CD3, oxo groups, -NH2, -COOH, -C(O)OC 1-6 Alkyl, -OH, 5-7 membered heteroaryl, 4-7 membered heterocyclic and C 3-6 Cycloalkyl; or two substituents located on the same or adjacent ring atoms and the ring atoms attached to them together to form: C 3-7 Carbon rings or 3-7 membered heterocycles, wherein the C 3-7 The carbocyclic ring or 3-7 membered heterocycle is optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene (=CH2), C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-3 Alkyl groups, halogens, -CN, CF3, -NH2, -OH, or oxo groups; *Optionally substituted amino groups, wherein one or more substituents are independently selected from: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Hydroxyalkyl, C 1-6 Alkyl groups, halogens, -CN, -NO2, -CF3, -NH2, and -OH; the substituents may optionally be substituted with one or more halogens; R1, R1 ’ and R2 ’ Independently selected from the following groups: hydrogen, deuterium, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, deuterated C 1-6 Alkyl, deuterated C 2-6 alkenyl, deuterated C 2-6 alkynyl group, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy groups, halogens, halogenated C 1-6 Alkyl, -CN, -N(C) 1-6 Alkyl groups (-2), CF3, -NH2, and -OH; R2 is selected from: hydrogen, -NH2, C 1-6 Alkyl, C 1-6 Alkoxy, -NH2, -N(C) 1-6 Alkyl)2、-NH-C 1-6 Alkyl group, -NHC(O)-C 1-6 Alkyl, -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-(4-7 membered heterocyclic group), -NHC(O)-(4-7 membered heteroaryl group), -NHC(O)-O-NH-C 1-6 Alkyl group, -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 1-6 Alkyl), -NHC(O)-N(C) 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 1-6 Alkyl, -NHC(O)-OC 3-7 Cycloalkyl, -NHC(O)-N(C) 1-6 Alkyl)2、-NHC(O)-N(C 3-7 cycloalkyl)2、-NHC(O)-O-CH2-C 3-7 cycloalkyl, -NHSO2-C 1-6 Alkyl group, -NHSO2-NH-C 1-6 Alkyl groups and -NHSO2-N(C 1-6 Alkyl groups (2), which are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Alkoxy, C 3-7 Cycloalkyl, deuterium, halogen, -CN, CF3, -NH2 and -OH; the heterocyclic or heteroaryl group contains one or more heteroatoms each independently selected from O, S and N; R a and R a ’ Each is independently selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne, halogen, -CN, CF3, -NH2, -OH, -C 1-3 Alkoxy, C 3-7 cycloalkyl and C 3-7 Heterocyclic groups; R b and R b ’ Each is independently selected from: hydrogen, C 1-6 Alkyl, C 2-6 alkenyl, C 3-7 cycloalkyl, C 5-10 Aryl and C 5-10 Heteroaryl groups, which are optionally substituted by one or more substituents, each independently selected from the following group: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 3-7 Cycloalkyl, halogen, -CN, CF3, -NH2, -OH and -C 1-3 alkoxy; or R b and R b ’ Together with the carbon atoms bonded to them, they form: C 3-8 Carbon ring, C 5-9 Spiral ring, C 5-10 Aromatic rings or C 5-10 Aromatic heterocycles, which are optionally substituted by one or more substituents, each independently selected from the group consisting of: methylene, C... 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, -C 1-3 Alkoxy, halogen, -CN, CF3, -NH2, and -OH; the substituents may optionally be substituted with one or more halogens; L is absent or selected from -O- and -NH-; m is 0, 1, or 2; and n is 0 or 1.

2. The compound according to claim 1, or its enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, characterized in that, L does not exist, m is 0, n is 1, and the compound is represented by formula (Ia): in Part A1 is selected from the group consisting of 5-12-membered heteroaryl groups containing one or more nitrogen atoms, wherein the heteroaryl group is optionally substituted by one or more substituents independently selected from the group consisting of halogen, hydroxyl, cyano, -NH2, C. 1-6 Alkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl groups and 4-7 membered heterocycloalkyl groups containing one or more heteroatoms each independently selected from O, S, and N; Part B1 is selected from: C 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12 Aryl, or 6-12-membered heteroaryl, optionally substituted with one or more halogens, wherein the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups. Part of C1 is selected from: 4-9 membered heterocyclic groups containing one or more N atoms, wherein the heterocyclic group is optionally substituted by one or more substituents independently selected from the group consisting of: halogen, methylene, and C. 1-6 Alkyl groups, wherein the substituents are optionally substituted with one or more halogens, or formed by two substituents on the same ring atom and the ring atom to which they are attached: C 3-7 Carbon rings; R1, R1 ’ and R2 ’ Independently selected from the following groups: hydrogen, CF3, and C 1-6 alkyl; R2 is selected from: -NH2, -NHC(O)-C 1-6 Alkyl group, -NHC(O)-NH-(C 1-6 alkyl), -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-N(C 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 3-7 Cycloalkyl, -NHC(O)- (4-7 membered heterocyclic) and -NHC(O)- (4-7 membered heteroaryl), each optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl and C 1-6 Alkoxy group; the heterocyclic group contains one or more heteroatoms, each independently selected from O, S and N; R b1 and R b1 ’ Each is independently selected from: C 3-7 cycloalkyl or with one or more C 3-7 Cycloalkyl-substituted C 2-6 alkenyl; or R b1 and R b1 ’ And together with the carbon atoms to which they are attached, they form: optionally by one or more C atoms. 1-6 alkyl or halogen-substituted C 3-8 Carbon ring.

3. The compound according to claim 1, or its enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, characterized in that, L does not exist, m is 0, n is 1, and the compound is represented by formula (Ib): in Part D is selected from the following group: C 1-6 Alkyl, C 3-7 cycloalkyl, -OC 3-7 cycloalkyl, -O-CH2-C 3-7 Cycloalkyl, -O-CH2-5-7-membered heteroaryl, -NH(C 1-6 alkyl), -N(C) 3-7 cycloalkyl), -N(C) 1-6 Alkyl)2, -N(C 1-6 Alkyl)(C 3-7 cycloalkyl), C 6-12 Aryl, 5-12 membered heteroaryl, and 4-7 membered heterocyclic groups, optionally substituted by one or more substituents independently selected from the group consisting of: H, C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, C 3-7 Cycloalkyl, 4-7 membered heterocyclic alkyl, halogen, halogenated C 1-6 Alkyl, C 1-6 Cyanoalkyl, oxo, -CN, -NH2, -OH, phenyl, -NH(C) 1-6 Alkyl group), -(CH2) q -NH(CO)-(C 1-4 alkyl) and -NHS(O)2(C 1-6 Alkyl); q is 0, 1, 2, 3, or 4; or, two substituents located on the same or adjacent ring atoms and the ring atoms to which they are attached together form: substituted or unsubstituted C 5-7 The heterocyclic group is a 5-7 membered heterocyclic group, either substituted or unsubstituted; the heterocyclic alkyl, heterocyclic and heteroaryl groups contain one or more heteroatoms independently selected from O, S and N; Part B2 is selected from: C 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms each independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens; the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups; Some C2s are selected from the following groups: The amino group, optionally substituted with one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, halogen or halogenated C 1-6 alkyl; A 4-9 membered heterocyclic group containing one or more heteroatoms selected from N, O, and S, wherein the heterocyclic group is optionally substituted by one or more substituents independently selected from the group consisting of methylene, C, and S. 1-6 Alkyl, C 2-6 alkenyl, C 1-6 Alkoxy, -SC 1-6 Alkyl, C 3-6 carbonyl group, -OC 3-6 Cycloalkyl, -NH2, CF3, CD3, CN, oxo, =NH, -N(C 1-6 alkyl)2、-NH(C 1-6 alkyl), -NHC(O)-(C 1-6 Alkyl), -NHS(O)2-(C 1-6 Alkyl), -NHS(O)2-(C 3-6 cycloalkyl), C 1-3 Hydroxyalkyl, -C(O)-C 1-6 Alkyl, halogen, -OH, -COOH, 5-7 membered heterocyclic groups, and 5-7 membered heteroaryl groups; the substituents may optionally be replaced by one or more halogens, deuterium, CN, CF3, oxo groups, CD3, -OH, -COOH, or -C(O)OC. 1-6 Alkyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-3 Alkoxy, 5-7 aryl and C 3-6 Cycloalkyl substitution; optionally, two substituents located on the same or adjacent ring atoms and the ring atoms to which they are attached together form: C 3-7 A carbon ring, or a 3-7 membered heterocycle containing one or more heteroatoms selected from N, O and S, or a 5-7 membered heteroaryl; The C 3-7 The carbocyclic ring or 3-7 membered heterocycle is optionally substituted by one or more substituents independently selected from the group consisting of: halogen, oxo group, C. 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 2-6 alkynyl group, C 1-3 Alkoxy groups, -CN, -CF3, -NH2, and -OH; R1, R1 ’ and R2 ’ Independently selected from the following groups: hydrogen, CF3, and C 1-6 alkyl; R2 is selected from: -NHC(O)-OC 1-6 Alkyl group, -NHC(O)-C 1-6 Alkyl group, -NHC(O)-NH-(C 1-6 alkyl), -NHC(O)-(C 3-7 cycloalkyl), -NHC(O)-NH-(C 3-7 cycloalkyl), -NHC(O)-N(C 1-6 Alkyl)(C 3-7 cycloalkyl), -NHC(O)-OC 3-7 Cycloalkyl, -NHC(O)-O-CH2-C 3-7 Cycloalkyl, -NHC(O)- (4-7 membered heterocyclic groups), -NHC(O)-N(C 1-6 Alkyl)2、-NHC(O)-N(C 3-7 Cycloalkyl)2 and -NHC(O)- (4-7-membered heteroaryl), each optionally substituted by one or more substituents independently selected from the group consisting of: deuterium, halogen, C 1-6 Alkyl, C 2-6 alkenyl, halogenated C 2-6 alkenyl, C 1-6 Alkoxy and C 3-7 Cycloalkyl; the heterocyclic or heteroaryl group comprises one or more heteroatoms independently selected from O, S, and N. R b2 and R b2 ’ Each is independently selected from: C 3-7 cycloalkyl; or, R b2 and R b2 ’ And together with the carbon atoms they are attached to form: C 3-8 A carbocyclic or 5-7 membered heterocycle, optionally substituted by one or more substituents independently selected from the group consisting of: methylene, C... 2-6 alkenyl, C 1-6 Alkyl or halogen; the substituent is optionally substituted with one or more halogens; or, two substituents located on the same or adjacent ring atoms and the ring atoms to which they are attached together form: C 3-6 A carbocyclic ring or a 4-7 membered heterocyclic group; said carbocyclic ring and heterocyclic group are optionally substituted by one or more substituents, each independently selected from the group consisting of: C 2-6 alkenyl, C 1-6 Alkyl or halogen; the substituent may optionally be replaced by one or more halogens. R c and R c ’ Each element is independently selected from: hydrogen, deuterium, and carbon. 1-3 Alkyl, C 1-3 Alkyl groups, halogens, -CN, -CF3, -CD3, -NH2, and -OH; p is 0 or 1; Preferably, R2 ’ Is it CF3 or C? 1-3 Alkyl groups; and / or Part B2 is selected from: phenyl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N atoms, wherein the phenyl is substituted with two halogens, the 6-membered heteroaryl group is unsubstituted or substituted with one halogen, and the 6-membered heterocyclic group is unsubstituted or substituted with one halogen or oxo group; and / or Part of C2 is selected from: 5-9 membered heterocyclic groups containing one or more N atoms, said heterocyclic group being substituted with a =CF2 and optionally substituted with one or more halogens or C. 1-6 Alkyl substitution, or two substituents on the same or adjacent ring atoms and the ring atoms to which they are attached, together form: C 3-7 Carbon rings; and / or R b2 and R b2 'And together with the carbon atoms they are attached to form: C atoms replaced by a =CF2' 5-8 Carbon ring.

4. The compound according to claim 1, or its enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, characterized in that, L does not exist, m is 0, n is 0, and the compound is represented by formula (Ic): in Part of D1 is selected from the following group: C 6-12 Aryl and 5-12 heteroaryl groups, which are optionally substituted by one or more substituents independently selected from the group consisting of: C 1-6 Alkyl, C 1-6 Hydroxyalkyl, C 1-6 Alkoxy, halogen, -CN, -NH2, and -OH; the heterocyclic alkyl and heteroaryl groups contain one or more heteroatoms independently selected from O, S, and N; Part of B3 is selected from: C 6-12 aryl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-12-membered heteroaryl group containing one or more N heteroatoms, wherein the C 6-12 The aryl or 6-12-membered heteroaryl group is optionally substituted with one or more halogens, wherein the 6-membered heterocyclic group is optionally substituted with one or more halogens or oxo groups; Part of C3 is selected from the following group: arbitrarily selected by one or more Cs 1-6 Alkyl-substituted amino groups, wherein the C 1-6 The alkyl group may optionally be substituted with one or more CF3 groups; the heterocyclic group contains one or more N atoms and is a 4-9 membered heterocyclic group, said heterocyclic group may optionally be substituted with one or more halogens, methylene groups, or C atoms. 2-6 Alkenyl substitution, the methylene and C 2-6 Alkenyl groups are optionally converted to one or more halogens or C. 1-6 Alkyl substitution, or two substituents located on the same ring atom and the ring atom attached to them, together form: C 3-7 Carbon rings; R2 and R2 ’ Independently selected from the following groups: hydrogen, CF3, and C 1-6 alkyl; R b3 and R b3 ’ Each is independently selected from: hydrogen, C 3-7 cycloalkyl or with one or more C 3-7 Cycloalkyl-substituted C 2-6 alkenyl; or R b3 and R b3 ’ And together with the carbon atoms to which they are attached, they are formed: optionally by one or more methylene groups, C 1-6 Alkyl, C 2-6 Alkenyl or halogen-substituted C 3-8 A carbon ring; the substituents are optionally substituted with one or more halogens; R c and R c ’ Each element is independently selected from: hydrogen, deuterium, and carbon. 1-3 Alkyl, C 1-3 Alkyl groups, halogens, -CN, -CF3, -CD3, -NH2, and -OH; p is 0 or 1; Preferably, Part B3 is selected from: phenyl, a 6-membered heterocyclic group containing one or more heteroatoms independently selected from O, S, and N, or a 6-membered heteroaryl group containing one or more N atoms, wherein the phenyl is substituted with two halogens, the 6-membered heteroaryl group is unsubstituted or substituted with one halogen, and the 6-membered heterocyclic group is unsubstituted or substituted with one halogen or oxo group; and / or The C3 is selected from: 5-9 membered heterocyclic groups containing one or more N atoms, said heterocyclic group being substituted with a =CF2 and optionally substituted with one or more halogens or C atoms. 1-6 Alkyl substitution, or two substituents on the same ring atom and the ring atom attached to them, together form: C 3-7 Carbon rings; and / or R b3 and R b3 'And together with the carbon atoms they are attached to form: C atoms substituted with a =CF2' 5-8 Carbon ring.

5. The compound according to any one of claims 1-4, or its enantiomers, stereoisomers, solvates, or pharmaceutically acceptable salts thereof, characterized in that, The compounds are selected from the group consisting of:

6. A pharmaceutical composition comprising a compound of any one of claims 1-5 and a pharmaceutically acceptable excipient.

7. A method for treating a disease or ailment of a subject in need, characterized in that, The disease or condition includes administering to a subject a therapeutically effective amount of any one of claims 1-5, or an enantiomer, stereoisomer, solvate, or pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 6, wherein the disease or condition is selected from the group consisting of inflammatory diseases, proliferative diseases, and autoimmune diseases.

8. The method according to claim 7, characterized in that, The disease or condition described is selected from one or more of the following: plaque psoriasis, guttate psoriasis, reversal psoriasis, pustular psoriasis, erythrodermic psoriasis, psoriatic arthritis, ankylosing spondylitis, hidradenitis suppurativa, palmoplantar psoriasis, airway inflammation, ankylosing spondylitis, asthma, rheumatoid arthritis, osteoarthritis, spondyloarthritis, bone erosion, intraperitoneal abscess and adhesions, IBD (inflammatory bowel disease), Crohn's disease, allograft rejection, psoriasis, psoriatic arthritis, certain types of cancer, blood Tube formation, atherosclerosis and multiple sclerosis, erythema, reactions to allergen exposure, Helicobacter pylori-associated gastritis, bronchial asthma, asthma, allogeneic graft rejection (e.g., kidney), systemic lupus erythematosus, lupus nephritis, Behcet's disease, ulcerative colitis, rheumatoid arthritis (RA), inflammatory bowel disease, Wegener's granulomatosis, sarcoidosis, systemic sclerosis, insulin-dependent diabetes mellitus, septic shock syndrome, Alzheimer's disease, inflammatory eye disease, uveitis and non-infectious uveitis.

9. The use of any compound of claims 1-5, or an enantiomer, stereoisomer, solvate, or pharmaceutically acceptable salt thereof, or the use of the pharmaceutical composition of claim 6 in the preparation of a medicament for the prevention or treatment of a disease or condition, wherein the disease or condition is selected from the group consisting of inflammatory diseases, proliferative diseases, and autoimmune diseases.

10. The use according to claim 9, characterized in that, The disease or condition described is selected from one or more of the following: plaque psoriasis, guttate psoriasis, reversal psoriasis, pustular psoriasis, erythrodermic psoriasis, psoriatic arthritis, ankylosing spondylitis, hidradenitis suppurativa, palmoplantar psoriasis, airway inflammation, ankylosing spondylitis, asthma, rheumatoid arthritis, osteoarthritis, spondyloarthritis, bone erosion, intraperitoneal abscess and adhesions, IBD (inflammatory bowel disease), Crohn's disease, allograft rejection, psoriasis, psoriatic arthritis, certain types of cancer, blood Tube formation, atherosclerosis and multiple sclerosis, erythema, reactions to allergen exposure, Helicobacter pylori-associated gastritis, bronchial asthma, asthma, allogeneic graft rejection (e.g., kidney), systemic lupus erythematosus, lupus nephritis, Behcet's disease, ulcerative colitis, rheumatoid arthritis (RA), inflammatory bowel disease, Wegener's granulomatosis, sarcoidosis, systemic sclerosis, insulin-dependent diabetes mellitus, septic shock syndrome, Alzheimer's disease, inflammatory eye disease, uveitis and non-infectious uveitis.

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