Polypeptide inhibitors, their production methods and applications

Abstract

The present invention relates to polypeptide inhibitors, their preparation methods and applications. In particular, the present invention relates to compounds represented by general formula (I), their preparation methods and pharmaceutical compositions containing the compounds, and their use as inhibitors in the treatment of multiple inflammatory and autoimmune diseases and cancer, wherein each substituent in general formula (I) is as defined in the specification.

Description

[Technical Field]

[0001] This application claims priority to Chinese Patent Application No. 2022115304381, filed November 30, 2022. This application incorporates all of the above Chinese patent applications by reference. This application claims priority from Chinese Patent Application No. 2023102062995, filed on March 3, 2023. This application incorporates all of the above Chinese patent applications by reference. This application claims priority from Chinese Patent Application No. 2023102618188, filed on March 15, 2023. This application incorporates all of the above Chinese patent applications by reference. This application claims priority from Chinese Patent Application No. 202310349519X, filed on March 31, 2023. This application incorporates all of the above Chinese patent applications by reference. This application claims priority from Chinese Patent Application No. 2023111444895, filed on September 5, 2023. This application incorporates all of the above Chinese patent applications by reference.

[0002] The present invention belongs to the field of biomedicine, and specifically relates to polypeptide inhibitors, their preparation methods and applications. [Background technology]

[0003] The cytokine IL-23 plays an important role in human innate and adaptive immunity. IL-23 induces the expression of proinflammatory cytokines in lymphocytes, most notably helper T cells (TH17), innate lymphoid cells (ILCs), and gamma-delta T cells. The IL-23 heterodimeric receptor consists of two subunits, IL-23R and IL-12Rβ1. IL-23R is a subunit unique to the IL-23 pathway. IL-12Rβ1 is shared with the IL-12 receptor. Similarly, the IL-23 cytokine consists of two subunits, p19 and p40. The p19 subunit is unique to IL-23, and the p40 subunit is shared with IL-12.

[0004] IL-23 provides the necessary conditions for the generation and survival of Th17 cells. At the same time, a large amount of evidence from preclinical models and clinical practice has shown that Th17 cells play a crucial role in the pathology of many autoimmune diseases, including inflammatory bowel disease, psoriasis vulgaris, rheumatoid arthritis, systemic lupus erythematosus (SLE), and multiple sclerosis (MS). Evidence from preclinical models and clinical trials has shown that blocking IL-23 signaling is effective in treating autoimmune diseases. However, the shared ligand and receptor subunit properties of the IL-23 and IL-12 pathways imply a more complex biological process. Furthermore, data on tumorigenesis, infection predisposition, and autoimmune disorders suggest that IL-23 blockade may offer therapeutic advantages in terms of efficacy and safety compared with IL-12 blockade. Summary of the Invention [Problem to be solved by the invention]

[0005] Limitations of antibody therapy include high production costs, poorer stability, higher demands on transportation, storage, use, and production, and typically requires infusion or injection during treatment, which poses challenges to patient compliance. Antibody immunosuppressive therapy is generally systemic, and treated tuberculosis patients face the risk of relapse and other serious infections. Therefore, when deploying anti-TNF or anti-IL23 therapy in many developing countries, it is necessary to exclude the high proportion of tuberculosis or hepatitis B (HBV) patients in the latent stage. Systemic antibody therapy has a very long half-life, resulting in the production of anti-drug antibodies (ADAs) that neutralize the antibody drug, resulting in reduced therapeutic efficacy. Intermittent dosing of anti-TNF antibodies significantly increases the likelihood of ADA development, which increases the treatment risk for patients.

[0006] Therefore, the object of the present invention is to provide molecules that specifically block IL-23 heterodimeric receptors by blocking IL-23R, compositions containing these molecules, methods for screening these molecules, and methods for using these molecules to treat multiple inflammatory and autoimmune diseases and cancers. These molecules should have good affinity and specificity and be distributed to inflammatory disease tissues without significantly affecting systemic immunity. [Means for solving the problem]

[0007] An object of the present invention is to provide a compound represented by general formula (IA), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the structure of the compound represented by general formula (IA) is as follows: [ka] where: R A is -(CH2) mA -R AA is selected from R AA is -NH2, -N + H3, -N + (CH3)3 or -NH-C(O)-CH3; m A is selected from integers from 1 to 6, Preferably, R A teeth, [ka] is selected from R B is -(CH2) mB -R BB is selected from R BB is -NR 1 R 2 , -N + (R 1 )2R 2 or -NH-C(O)-R 1 is selected from R 1 is H, alkyl group, [ka] or amino acids, preferably R 1 is selected from H, an alkyl group, R 2 is H, alkyl group, [ka] or an amino acid, R a is H, alkyl group, [ka] or an amino acid, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; Preferably, R BB is -NH2, -N + H3, -N + (CH3)3 or -NH-C(O)-CH3; m B is selected from integers from 1 to 6, Preferably, R B teeth, [ka] is selected from R 3 represents a hydroxy group, an alkyl group, or -NR 6 R 7 or an amino acid, R 6 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R7 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c is H, alkyl group, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from an amino acid; X'7 is selected from amino acids; R 4 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 is H, alkyl group, -CH2C(O)NH2, [ka] or amino acids, preferably [ka] is selected from R b is H, alkyl group, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n1 and n2 are each independently selected from integers of 0 to 12; t2 to t9 are each independently selected from integers of 0 to 12; m4 to m22 are each independently selected from integers of 0 to 24.

[0008] An object of the present invention is to provide a compound represented by general formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the structure of the compound represented by general formula (I) is as follows: [ka] where: R 1 and R 2 are each independently H, an alkyl group, [ka] or an amino acid, R a is H, alkyl group, [ka] or an amino acid, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; Preferably, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; R 3 represents a hydroxy group, an alkyl group, or -NR 6 R 7 or an amino acid, R 6 and R 7 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c is H, alkyl group, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X6 is selected from amino acids; X7 is selected from an amino acid; Preferably, R 3 represents a hydroxy group, an alkyl group, or -NR 6 R 7 or an amino acid, R 6 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 7 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c is H, alkyl group, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from an amino acid; X'7 is selected from amino acids; R 4 and R 5 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R b is H, alkyl group, [ka] or an amino acid, R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X3 is selected from amino acids; X4 is selected from amino acids; Preferably, R 4 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 is H, alkyl group, -CH2C(O)NH2, [ka] or amino acids, preferably [ka] is selected from R b is H, alkyl group, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n to n2 are each independently selected from integers of 0 to 12; t to t9 are each independently selected from integers of 0 to 12; m to m21 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; The compound is [ka] isn't it.

[0009] Another object of the present invention is to provide a compound represented by general formula (IV), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the structure of the compound represented by general formula (IV) is as follows: [ka] where: X 1 is selected from amino acids, X 2 is selected from amino acids, R 1 and R 2 are each independently H, an alkyl group, [ka] or an amino acid, R a is H, alkyl group, [ka] or an amino acid, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; R 3 represents a hydroxy group, an alkyl group, or -NR 6 R 7 or an amino acid, R 6 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 7 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c is H, alkyl group, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from an amino acid; X'7 is selected from amino acids; R 4 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 is H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R b is H, alkyl group, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n to n2 are each independently selected from integers of 0 to 12; t to t9 are each independently selected from integers of 0 to 12; m to m21 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; The compound is [ka] isn't it.

[0010] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 1 and R 2 are each independently H, an alkyl group, [ka] or an amino acid, R a H, alkyl groups, [ka] or an amino acid, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; Preferably, Raa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; R 3 is selected from a hydroxy group; R 4 and R 5 are each independently selected from H, an alkyl group, and —CHC(O)NH; n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24.

[0011] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 1 and R 2 is selected from H, R 3 hydroxy group, alkyl group, -NR 6 R 7 or an amino acid, R 6 and R 7 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c H, alkyl groups, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X6 is selected from amino acids; X7 is selected from amino acids; Preferably, R 3 hydroxy group, alkyl group, -NR 6 R 7 or an amino acid, R 6 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 7 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c H, alkyl groups, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from amino acids; X'7 is selected from an amino acid; R 4 and R 5 are each independently selected from H, an alkyl group, and —CHC(O)NH; n2 is selected from integers from 0 to 12; t6 to t9 are each independently selected from integers of 0 to 12; m13 to m21 are each independently selected from integers of 0 to 24.

[0012] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 1 and R 2 is selected from H, R 3 is selected from a hydroxy group; R 4 and R 5 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R b H, alkyl groups, [ka] or an amino acid, R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X3 is selected from amino acids; X4 is selected from amino acids; Preferably, R 4 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 H, alkyl group, -CH2C(O)NH2, [ka] or amino acids, preferably [ka] is selected from R b H, alkyl groups, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers of 0 to 24.

[0013] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is further as shown in general formula (II): [ka] where: R 1 and R 2 are each independently H, an alkyl group, [ka] or an amino acid, R aH, alkyl groups, [ka] or an amino acid, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; Preferably, R aa is an alkyl group, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; R 4 is selected from H or an alkyl group; n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24.

[0014] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is further represented by general formula (III): [ka] where: R 3 hydroxy group, alkyl group, -NR 6 R 7 or an amino acid, R 6 and R 7 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c H, alkyl groups, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X6 is selected from amino acids; X7 is selected from amino acids; Preferably, R 3 hydroxy group, alkyl group, -NR 6 R 7 or an amino acid, R 6 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 7 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c H, alkyl groups, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from amino acids; X'7 is selected from an amino acid; R 4 is selected from H or an alkyl group; n2 is selected from integers from 0 to 12; t6 to t9 are each independently selected from integers of 0 to 12; m13 to m21 are each independently selected from integers of 0 to 24.

[0015] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is further represented by general formula (V): [ka] where: X 1 is selected from amino acids, X 2 is selected from amino acids, R 4 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R b H, alkyl groups, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X'2 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 12; The compound is [ka] It is characterized by not being

[0016] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is X 1 But, Thr, [ka] is selected from, preferably [ka] It is characterized in that:

[0017] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is X 2 but, [ka] is selected from, preferably [ka] It is characterized in that:

[0018] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is further represented by general formula (IV): [ka] where: R 4 is selected from H or an alkyl group; R 5 is an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R b H, alkyl groups, [ka] or amino acids R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X3 is selected from amino acids; X4 is selected from amino acids; Preferably, R 4 H, alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R 5 H, alkyl group, -CH2C(O)NH2, [ka] or amino acids, preferably [ka] is selected from Rb H, alkyl groups, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb is an alkyl group, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers of 0 to 24.

[0019] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 1 But H, C 1-6 alkyl groups, [ka] or an amino acid, R a But H, C 1-6 alkyl groups, [ka] or an amino acid, R aa But C 1-6 alkyl groups, [ka] or an amino acid, X1 is selected from amino acids; Preferably, R aa But C 1-6 alkyl groups, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24.

[0020] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 2 But H, C 1-6 alkyl groups, [ka] or an amino acid, R a But H,C 1-6 alkyl groups, [ka] or an amino acid, R aa But C 1-6 alkyl groups, [ka] or an amino acid, X1 is selected from amino acids; Preferably, R aa But C 1-6 alkyl groups, [ka] or an amino acid, X1 is selected from amino acids; X'1 is selected from amino acids; n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24.

[0021] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 3 is a hydroxy group, C 1-6 Alkyl group, -NR 6 R 7 or an amino acid, R 6 and R 7 are each independently H, an alkyl group, -CH2C(O)NH2, [ka] or an amino acid, R c H, alkyl groups, [ka] or an amino acid, R cc is an alkyl group, [ka] or an amino acid, X5 is selected from amino acids; X6 is selected from amino acids; X7 is selected from amino acids; Preferably, R 3 is a hydroxy group, C 1-6 Alkyl group, -NR 6 R 7 or an amino acid, R 6 But H, C 1-6alkyl groups, -CH2C(O)NH2, [ka] or an amino acid, R 7 But H, C 1-6 alkyl groups, -CH2C(O)NH2, [ka] or an amino acid, R c But H, C 1-6 alkyl groups, [ka] or an amino acid, R cc But C 1-6 alkyl groups, [ka] or an amino acid, X5 is selected from amino acids; X'5 is selected from amino acids; X6 is selected from amino acids; X'6 is selected from amino acids; X7 is selected from amino acids; X'7 is selected from an amino acid; n2 is selected from integers from 0 to 12; t6 to t9 are each independently selected from integers of 0 to 12; m13 to m21 are each independently selected from integers of 0 to 24.

[0022] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 4 But H, C 1-6 alkyl groups, -CH2C(O)NH2, [ka] or an amino acid, R b But H, C 1-6 alkyl groups, [ka] or an amino acid, R bb But C 1-6 alkyl groups, [ka] or an amino acid, X2 is selected from amino acids; X3 is selected from amino acids; X4 is selected from amino acids; Preferably, R 4 But H, C 1-6 alkyl groups, -CH2C(O)NH2, [ka] or an amino acid, R b But H, C 1-6 alkyl groups, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb But C 1-6 alkyl groups, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers of 0 to 24.

[0023] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is R 5 But H, C 1-6 alkyl groups, -CH2C(O)NH2, [ka] or an amino acid, R b But H, C 1-6 alkyl groups, [ka] or an amino acid, R bb But C 1-6 alkyl groups, [ka] or an amino acid, X2 is selected from amino acids; X3 is selected from amino acids; X4 is selected from amino acids; Preferably, R 5 But H, C 1-6 alkyl groups, -CH2C(O)NH2, [ka] or amino acids, more preferably [ka] is selected from R b But H, C 1-6 alkyl groups, [ka] or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R bb But C 1-6 alkyl groups, [ka] or an amino acid, X2 is selected from amino acids; X'2 is selected from amino acids; X3 is selected from amino acids; X'3 is selected from amino acids; X4 is selected from amino acids; X'4 is selected from amino acids; n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers of 0 to 24.

[0024] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0025] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0026] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from Or, [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0027] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from Or, [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0028] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from, preferably [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0029] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from, preferably [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0030] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0031] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from Or, [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0032] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is [ka] but, [ka] is selected from Or, [ka] but, [ka] The present invention is characterized in that the compound is selected from the group consisting of:

[0033] In a preferred embodiment of the invention, the amino acid is Gly, Ala, Val, Leu, Ile, Phe, Trp, Tyr, Asp, His, Asn, Glu, Lys, Gln, Met, Arg, Ser, Thr, Cys or Pro, Preferably, the amino acid is Gln, More preferably, the amino acid is Gln or Glu.

[0034] In a further embodiment of the invention, n is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12; n1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; n2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t4 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t5 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t6 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t7 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t8 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t9 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; m is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24; m1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m4 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m5 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m6 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m7 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m8 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m9 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m10 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m11 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m12 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m13 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m14 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m15 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m16 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m17 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m18 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m19 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m20 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m21 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24.

[0035] m22 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24.

[0036] In a preferred embodiment of the present invention, the compound, its stereoisomer or its pharmaceutically acceptable salt is a compound in which the amino acid is selected from the group consisting of Gly, Ala, Val, Leu, Ile, Phe, Trp, Tyr, Asp, His, Asn, Glu, Lys, Gln, Met, Arg, Ser, Thr, Cys, Pro, [ka] and Preferably, the amino acid is Glu, Thr, [ka] It is characterized in that:

[0037] The present invention further provides a method for preparing the above compound, its stereoisomer or its pharmaceutically acceptable salt, which is based on solid phase or liquid phase synthesis, Preferably, the synthesis method comprises: Step 1) synthesizing a resin peptide based on a solid phase synthesis method; Step 2) of cleaving the resin peptide obtained in step 1) to obtain a polypeptide intermediate; Step 3) of condensing the polypeptide intermediate with the side chain; and step 4) of removing the protecting groups from the peptide segment obtained in step 3) and then forming a ring to obtain the final product, More preferably, the synthesis method comprises: Step 1) synthesizes a peptide on a resin based on the solid-phase synthesis method of the Fmoc method and end-caps it with acetic anhydride; Step 2) of cleaving the resin peptide obtained in step 1) to obtain a polypeptide intermediate; Step 3) of subjecting the polypeptide intermediate and the side chain to a condensation reaction using a coupling agent; and step 4) of removing the protecting groups from the peptide segment obtained in step 3) and then oxidizing the disulfide bond to form a ring to obtain the final product.

[0038] The present invention further relates to pharmaceutical compositions, which comprise a therapeutically effective amount of the compound, its stereoisomer, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0039] In a preferred embodiment of the invention, the pharmaceutical composition is selected from a tablet, a capsule, a liquid formulation or an injection.

[0040] In a preferred embodiment of the present invention, the pharmaceutical composition is an immediate release formulation or a sustained release formulation.

[0041] In some embodiments of the present invention, the compound, its stereoisomer or a pharmaceutically acceptable salt thereof may be administered in any convenient manner, such as, for example, oral, parenteral, buccal, sublingual, nasal, rectal, intrathecal or transdermal administration, and a correspondingly prepared pharmaceutical composition.

[0042] In some embodiments of the present invention, the compound, its stereoisomer, or its pharmaceutically acceptable salt may be formulated as a liquid or solid preparation, for example, a syrup, suspension, emulsion, tablet, capsule, powder, granule, or lozenge.

[0043] The present invention further relates to the use of the compound, its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the manufacture of an IL-23R inhibitor drug. The present invention further relates to the use of the compound, its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof in the treatment of inflammatory and autoimmune diseases and cancers, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, The present invention relates to an application in the manufacture of a medicament for treating Chediak-Higashi syndrome and Wiskott-Aldrich syndrome, pouchitis following proctocolectomy and ileal pouch-anal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholecithin, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), psoriasis vulgaris, psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetes mellitus.

[0044] The present invention relates to the use of said compounds, their stereoisomers or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof in the treatment of inflammatory, autoimmune diseases and cancer, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, and cancer. Further relates to applications in methods for treating Diack-Higashi syndrome and Wiskott-Aldrich syndrome, pouchitis following proctocolectomy and ileal pouch-anal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholecithin, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), psoriasis vulgaris, psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetic diseases.

[0045] The present invention further relates to the treatment of inflammatory, autoimmune diseases and cancer, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome and Wiskott-Aldrich syndrome, proctocolectomy and ileal pouch-anal anastomosis. and / or a method for treating and / or preventing pouchitis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), plaque psoriasis, plaque psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetes mellitus, which comprises administering to a patient a therapeutically effective amount of the compound, its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0046] The present invention further relates to the treatment of inflammatory, autoimmune diseases and cancer in mammals, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome, and Wiskott-Aldrich syndrome, and the treatment of colonic ulcers and rectal colonic resections and colonic ulcers following ileal pouch-anal anastomosis. and the like. The present invention relates to a method for treating a disease such as pouchitis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), plaque psoriasis, plaque psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetes mellitus caused by a compound of the present invention, which comprises administering to said mammal a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt, ester, prodrug, solvate, hydrate or derivative thereof.

[0047] In some embodiments, the methods include the use of a therapeutic agent in the treatment of inflammatory, autoimmune diseases and cancers, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Cheddia ulcerative colitis, and eosinophilic colitis. For Buck-Higashi syndrome and Wiskott-Aldrich syndrome, pouchitis following proctocolectomy and ileal pouch-anal anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), psoriasis vulgaris, psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetes mellitus, Preferably, the disease is inflammatory bowel disease, arthritis or psoriasis vulgaris. DETAILED DESCRIPTION OF THE INVENTION

[0048] Unless otherwise defined herein, scientific and technical terms used in this patent application have the meanings commonly understood by those skilled in the art. Generally, the nomenclature and techniques of such disciplines used in connection with chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described herein are those known and commonly used in the art.

[0049] The term "peptide" refers broadly to a sequence of two or more amino acids joined by peptide bonds. It should be understood that the term does not imply a specific length of an amino acid polymer, nor is it intended to imply or distinguish between polypeptides produced using recombinant technology, chemical or enzymatic synthesis, or naturally occurring polypeptides. The term peptide includes cyclic peptides.

[0050] The term "amino acid" refers to any and all amino acids and residues thereof, including naturally occurring amino acids (e.g., α-amino acids), unnatural amino acids, modified amino acids, synthetic amino acids, or rare amino acids, including both D- and L-amino acids. Natural amino acids include those found in nature, such as the 23 amino acids that combine into peptide chains to form the building blocks of many proteins. These stereoisomers are predominantly L-stereoisomers, although some D-amino acids occur in bacterial envelopes and some antibiotics. The 20 "standard" naturally occurring amino acids are alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). The "non-standard" naturally occurring amino acids are pyrrolysine (found in methanogens and other eukaryotes), selenocysteine ​​(present in many non-eukaryotes and the majority of eukaryotes), and N-formylmethionine (encoded by the start codon AUG in bacteria, mitochondria, and chloroplasts). "Non-natural" or "unnatural amino acids" are non-proteinogenic amino acids (i.e., amino acids that are not naturally encoded or found in the genetic code), either naturally occurring or chemically synthesized. Over 140 non-natural amino acids are known, and thousands of possible combinations are possible. Examples of "non-natural" amino acids are β-amino acids (β 3 and β 2), homoamino acids, proline and pyruvate derivatives, trisubstituted alanine derivatives, glycine derivatives, ring-substituted phenylalanine and tyrosine derivatives, linear amino acids, diamino acids, D-diamino acids, α-methyl amino acids, and N-methyl amino acids. Non-natural or unnatural amino acids further include modified amino acids. "Modified" amino acids include amino acids (e.g., natural amino acids) that have been chemically modified to include non-naturally occurring groups or chemical moieties on the amino acid. Preferably, non-natural amino acids according to the present invention are [ka] Including, but not limited to:

[0051] The term "alkyl group" refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched-chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, and 5-methylhexyl groups. , 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof.More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.

[0052] Any hydrogen atom described in the present invention may be substituted with its isotope, deuterium, and any hydrogen atom in the compounds of the examples of the present invention may also be substituted with a deuterium atom.

[0053] "Hydroxy" refers to an -OH group.

[0054] "THF" refers to tetrahydrofuran.

[0055] "MeOH" refers to methanol.

[0056] "DMF" refers to N,N-dimethylformamide.

[0057] "TFA" refers to trifluoroacetic acid.

[0058] "TEA" refers to triethylamine.

[0059] "MeI" refers to methyl iodide.

[0060] "DMA" refers to N,N-dimethylacetamide.

[0061] "Et2O" refers to ether.

[0062] "DCM" refers to dichloromethane;

[0063] "DMAP" refers to 4-dimethylaminopyridine.

[0064] "DCC" refers to dicyclohexylcarbodiimide.

[0065] "DCE" refers to 1,2 dichloroethane.

[0066] "DIEA" refers to N,N-diisopropylethylamine.

[0067] "NBS" refers to N-bromosuccinimide.

[0068] "NIS" refers to N-iodosuccinimide.

[0069] "Cbz-Cl" refers to benzyl chloroformate.

[0070] "Pd2(dba)3" refers to tri(dibenzylideneacetone)dipalladium.

[0071] "Dppf" refers to 1,1'-bisdiphenylphosphinoferrocene.

[0072] "HATU" refers to 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate.

[0073] "KHMDS" refers to potassium hexamethyldisilazane.

[0074] "LiHMDS" refers to lithium bistrimethylsilylamide.

[0075] "MeLi" refers to methyllithium.

[0076] "n-BuLi" refers to n-butyllithium.

[0077] "NaBH(OAc)3" refers to sodium triacetoxyborohydride.

[0078] A "pharmaceutical composition" is meant to contain a mixture of one or more compounds described herein, or physiologically / pharmaceutically acceptable salts or prodrugs thereof, with other chemical components, as well as other components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism and promote absorption of the active ingredients, thereby exerting their biological activity.

[0079] "Pharmaceutically acceptable salt" refers to a salt of a compound of the present invention, which is safe and effective when used in a mammalian body and possesses the desired biological activity. [Example]

[0080] The present invention will be further described below in conjunction with examples, but these examples are not intended to limit the scope of the present invention.

[0081] Example The structures of the compounds of the present invention were confirmed by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 nuclear magnetometer in deuterated dimethyl sulfoxide (DMSO-d), deuterated methanol (CD3OD), deuterated chloroform (CDCl3), or deuterated water (DO). The internal standard was tetramethylsilane (TMS).

[0082] Liquid chromatography-mass spectrometry (LC-MS) measurements were performed using a Thermo Scientific-LTQ-XL electrospray ionization mass spectrometer. HPLC measurements were performed using an Agilent 1260 high-pressure liquid chromatograph (Phenomenex Gemini C18, 4.6 x 150 mm, 5 μm chromatography column). Thin-layer chromatography silica gel plates were Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plates. TLC was performed using 0.15 mm to 0.20 mm silica gel plates, while thin-layer chromatography product separation and purification was performed using 0.4 mm to 0.5 mm silica gel plates. Column chromatography generally used Yantai Yellow Sea silica gel 200-300 mesh silica gel as the support.

[0083] The starting materials in the embodiments of the present invention are known and commercially available, or can be synthesized using or according to methods known in the art.

[0084] Unless otherwise specified, all reactions of the present invention are carried out under a dry nitrogen or argon atmosphere with continuous magnetic stirring, the solvents are dry solvents, and the reaction temperatures are in degrees Celsius.

[0085] The eluent system for silica gel column chromatography and the developer system for thin-layer chromatography used to purify the intermediates and compounds in the examples include A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, and C: dichloromethane and acetone system. The volume ratio of the solvents is adjusted according to the polarity of the compounds, and can also be adjusted by adding a small amount of alkaline or acidic reagents such as triethylamine and acetic acid.

[0086] Intermediate A [ka] [ka]

[0087] Step 1 To a solution of compound A1 (50 g, 0.19 mol) in DMF (350 mL) was added NaHCO3 (79.8 g, 0.95 mol) and MeI (80.9 g, 0.57 mol) at room temperature. The mixture was stirred at 30 °C for 16 hours, diluted with water, and extracted with ethyl acetate. The combined organic layer was washed with saturated brine, dried over Na2SO4, filtered, and the filtrate was concentrated to give compound A2 (55 g), which was used directly in the next step. LC / MS: [M+H] + -100=178.0.

[0088] Step 2 Compound A2 (10 g, 36.06 mmol) was dissolved in TFA / DCM (v:v=1:2, 100 mL) and stirred at room temperature for 2 hours. After concentrating the reaction mixture, compound A3 (10 g) was obtained, and the crude product was used directly in the next step. LC / MS: [M+H] + =178.1.

[0089] Step 3 To a solution of compound A1 (9.5 g, 36 mmol) in CH3CN (250 mL) at 0 °C, compound A3 (6.4 g, 36 mmol), DIEA (23.3 g, 180 mmol), and HATU (13.6 g, 36 mmol) were added. The reaction mixture was stirred at room temperature for 1 h, diluted with water, and extracted three times with ethyl acetate. The combined organic layer was washed with saturated brine, dried over Na2SO4, and filtered. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (0% to 5%, MeOH / DCM) to give compound A4 (15 g) in 98.7% yield. LC / MS: [M+H] + -100=323.2.

[0090] Step 4 Compound A4 (9.1 g, 21.54 mmol) was dissolved in HCl / dioxane (4N, 100 mL) and stirred at room temperature for 1 hour. After concentrating the reaction mixture, compound A5 (7 g) was obtained, and the crude product was used directly in the next step. LC / MS: [M+H] + =323.4.

[0091] Step 5 Compound A5 (6.88 g, 21.34 mmol), DIEA (13.79 g, 106.71 mmol), and HATU (8.05 g, 21.34 mmol) were added to a solution of (S)-4-(((benzyloxy)carbonyl)amino)-5-(tert-butoxy)-5-oxopentanoic acid (7.2 g, 21.34 mmol) in CH3CN (80 mL) at 0 °C, and the reaction mixture was stirred at room temperature for 16 h. The mixture was diluted with brine and extracted three times with ethyl acetate. The combined organic layer was washed with saturated brine, dried over Na2SO4, and filtered. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (0% to 5%, MeOH / DCM) to give compound A6 (7 g) in 51.1% yield. LC / MS: [M+H] + =642.6.

[0092] Step 6 To a solution of compound A6 (7 g, 10.91 mmol) in THF (150 mL) was added 10% Pd / C (700 mg), and the mixture was stirred at room temperature under a hydrogen atmosphere (1.5 atm) for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to give compound A7 (5.07 g) in a 91.6% yield. LC / MS: [M+H] + =508.4.

[0093] Step 7 Compound A7 (5.07 g, 10 mmol), DIEA (6.45 g, 50 mmol), and HATU (4.45 g, 13 mmol) were added to a solution of mono-tert-butyl hexadecanedioate (4.45 g, 12.99 mmol) in CH3CN (80 mL) at 0 °C. The reaction mixture was stirred at room temperature for 16 h, then diluted with brine and extracted three times with EtOAc. The combined organic layers were washed with saturated brine, dried over Na2SO4, and filtered. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (0% to 5%, MeOH / DCM) to give compound A8 (7.7 g) in 92.6% yield. LC / MS: [M+H] + =832.7, 1 H NMR(400MHz,CD3OD) δ4.24(dd,J=9.1,5.1Hz,1H),4.16(s,2H),4.00(s,2H),3.73(s,3H),3.69(dt,J=5.3,2 .7Hz,4H),3.66-3.62(m,4H),3.56(dt,J=8.4,5.5Hz,4H),3.44(t,J=5.4Hz,2H),3.37( t,J=5.5Hz,2H),2.29(t,J=7.7Hz,2H),2.25-2.16(m,4H),2.10(qd,J=7.6,5.3Hz,1H), 1.89(dt,J=16.5,7.5Hz,1H),1.64-1.52(m,4H),1.45(d,J=9.1Hz,18H),1.29(s,20H).

[0094] Step 8 At 0°C, a solution of compound A8 (4 g, 4.81 mmol) in THF (40 mL) was added with a solution of LiOH (0.23 g, 9.61 mmol) in water (10 mL), and the mixture was allowed to react at this temperature for 1 hour. 1N HCl was added to the reaction mixture to adjust the pH to 4, and the aqueous phase was extracted three times with EtOAc. The combined organic layer was washed with saturated brine, dried over Na2SO4, and then filtered. The filtrate was concentrated under reduced pressure to give compound A9 (4 g), and the crude product was used directly in the next step. LC / MS: [M+H] + =818.7.

[0095] Step 9 To a solution of compound A9 (3.5 g, 4.28 mmol) in CH3CN (40 mL) at 0 °C, DIEA (3.32 g, 25.67 mmol), HATU (1.94 g, 5.13 mmol), and 2-amino-N-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethyl)-N,N-dimethylethane-1-ammonium hydrochloride (1.92 g, 6.42 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound A10 (TFA salt, 2.16 g) in 43.7% yield. LC / MS: [M] + =1062.8.

[0096] Step 10 A solution of compound A10 (2 g, 1.88 mmol) in THF (20 mL) was added with aqueous LiOH (135 mg, 5.64 mmol, 10 mL) at 0°C and the mixture was allowed to react at this temperature for 1 hour. The reaction mixture was concentrated to remove THF, and the residue was adjusted to pH 7 by dropwise addition of 1N HCl. The mixture was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CHCN / 0.03% TFA / water) to give compound A11 (TFA salt, 0.7 g) in a 35.5% yield. LC / MS: [M] + =1048.8.

[0097] Step 11 To a solution of compound A11 (0.7 g, 0.67 mmol) in CH3CN (10 mL) at 0 °C, DIEA (517 mg, 4.0 mmol), HATU (302 mg, 0.8 mmol), and 2-amino-N-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethyl)-N,N-dimethylethane-1-ammonium hydrochloride (208 mg, 1.0 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound A12 (TFA salt, 820 mg) in 99.2% yield. LC / MS: [(M+H) / 2] + =620.4.

[0098] Step 12 To a solution of compound A12 (700 mg, 0.56 mmol) in THF (20 mL) was added 10% Pd / C (70 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CHCN / 0.03% TFA / water) to give compound intermediate A (TFA salt, 600 mg) in 95.1% yield. LC / MS: [M] + =1104.7. 1H NMR(400MHz,CD3OD) δ4.24(dd,J=9.2,5.2Hz,1H),4.02(d,J=14.0Hz,4H),3.94(s,2H),3.74(q,J=6.1Hz,4H),3.71-3.63(m, 14H),3.61-3.54(m,6H),3.48(dt,J=10.6,5.6Hz,4H),3.40-3.35(m,2H),3.22(s,6H),3.13(t,J=5.8Hz ,2H),2.73(s,3H),2.51(t,J=6.2Hz,2H),2.29(t,J=7.6Hz,2H),2.26-2.17(m,4H),2.11(ddd,J=15.6,1 0.3,6.6Hz,1H),1.95-1.82(m,1H),1.59(dt,J=22.3,7.1Hz,4H),1.45(d,J=9.0Hz,18H),1.29(s,20H).

[0099] Intermediate A1 [ka]

[0100] Step 1 L-Phenylglycine (3.35 g, 40.26 mmol) was dissolved in methanol (150 mL) and cooled to -78°C. A methanol solution (20 mL) of 6,6-dimethylbicyclo[3.1.0]hexan-3-one A1a (CAS: 13855-29-3, 5.0 g, 40.26 mmol) and a methanol solution (20 mL) of tert-butyl isocyanate were added dropwise in that order, and the mixture was gradually warmed to room temperature and allowed to react for 24 hours. The reaction mixture was concentrated, then dissolved in 200 mL of ether, and the mixture was triturated. Insoluble matter was removed by filtration. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (gradient elution with EA / PE system) to obtain compound (2S)-2-((3-(tert-butylaminoformyl)-6,6-dimethylbicyclo[3.1.0]hexan-3-yl)amino)-2-phenylacetic acid methyl ester A1b (10.5 g), with a yield of 70.0%. LC / MS: [M+H] + =373.3.

[0101] Step 2 Methyl (2S)-2-((3-(tert-butylaminoformyl)-6,6-dimethylbicyclo[3.1.0]hexan-3-yl)amino)-2-phenylacetate A1b (10.0 g, 26.84 mmol) and palladium hydroxide (1.13 g, 8.05 mmol) were dissolved in methanol (200 mL). The mixture was purged with hydrogen gas three times, and the hydrogen gas atmosphere was maintained while the reaction was carried out at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, and the filtrate was concentrated and then dissolved in ether (150 mL). The filtrate was extracted twice with 2N dilute hydrochloric acid (50 mL). The aqueous phases were combined, the pH was adjusted to 8 with solid potassium carbonate, and the mixture was extracted twice with ether (100 mL). The mixture was concentrated under reduced pressure to give 3-amino-N-(tert-butyl)-6,6-dimethylbicyclo[3.1.0]hexane-3-carboxamide A1c (4.9 g) in a yield of 81.4%. LC / MS: [M+H] + =225.2.

[0102] Step 3 3-Amino-N-(tert-butyl)-6,6-dimethylbicyclo[3.1.0]hexane-3-carboxamide A1c (4.9 g, 21.84 mmol) was dissolved in 6N hydrochloric acid (50 mL) and refluxed for 16 hours. The reaction mixture was cooled to room temperature, extracted with ether, washed, and the aqueous phase was collected and concentrated under reduced pressure to give 3-amino-6,6-dimethylbicyclo[3.1.0]hexane-3-carboxylic acid A1d (hydrochloride, 4.4 g) in 98% yield. LC / MS: [M+H] + =170.1.

[0103] Step 4 3-Amino-6,6-dimethylbicyclo[3.1.0]hexane-3-carboxylic acid A1d (hydrochloride, 4.4 g, 21.39 mmol) was dissolved in sodium bicarbonate solution (100 mL), and a dioxane solution (80 mL) of 9-fluorenylmethyl-N-succinimidyl carbonate (10.79 g, 32.09 mmol) was added. The mixture was allowed to react at room temperature for 1 hour. The reaction mixture was extracted twice with ethyl acetate (100 mL). The combined organic phase was washed with water. The aqueous phase was adjusted to pH 2 with 2N dilute hydrochloric acid and extracted twice with ethyl acetate (50 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was then purified by silica gel column chromatography (gradient elution with MeOH / DCM system) to obtain intermediate A1 3-((((9H-fluorenyl-9-yl)methoxy)carbonyl)amino)-6,6-dimethylbicyclo[3.1.0]hexane-3-carboxylic acid (5.6 g), with a yield of 66.9%. LC / MS: [MH] - =390.2.

[0104] Intermediate A2 [ka] By referring to the synthesis method of intermediate A1, 4-(difluoromethylene)cyclohexanone (CAS: 137780-61-1) and subsequent similar experimental procedures, intermediate A2 1-((((9H-fluorenyl-9-yl)methoxy)carbonyl)amino)-4-(difluoromethylene)cyclohexane-1-formic acid was obtained. LC / MS: [MH] - =412.1.

[0105] Intermediate A3 [ka]

[0106] Step 1 At 0°C, a solution of tetrahydrothiopyran-4-one A3a (50 g, 431.0 mmol) in methanol (30 mL) was added with ammonia in methanol (7 M, 300 mL) and stirred at 0°C for 3 h. Cyanotrimethylsilane (44.9 g, 452.6 mmol) was then added dropwise. After the addition was complete, the mixture was gradually warmed to room temperature and stirred for 1.5 h. The reaction mixture was concentrated to give a light brown oil, which was triturated with petroleum ether (300 mL) at 0°C to precipitate a solid. After filtration and drying, 4-aminotetrahydrothiopyran-4-nitrile A3b (61.5 g) was obtained, which was directly used in the next step without further purification. LC / MS: [M+H] + =143.1.

[0107] Step 2 A3b (61.5 g, 433.0 mmol) was dissolved in a mixture of dioxane (300 mL) and water (300 mL), and sodium carbonate (55.1 g, 519.7 mmol) was added. A solution of 9-fluorenylmethyl chloroformate (117.6 g, 454.7 mmol) in dioxane (100 mL) was added dropwise to the reaction mixture at 0 °C, and the mixture was allowed to warm to room temperature and stirred for 1 h. Water (400 mL) was added to the reaction mixture, which was triturated and filtered to give a white solid. The resulting solid was triturated with petroleum ether (300 mL), filtered, and dried to give (9H-fluorenyl-9-yl)methyl (4-cyanotetrahydro-2H-thiopyran-4-yl)carbamate A3c (124 g) in 79% yield. LC / MS: [M+H] + =365.2.

[0108] Step 3 (9H-Fluorenyl-9-yl)methyl (4-cyanotetrahydro-2H-thiopyran-4-yl)carbamate A3c (50.0 g, 137.4 mmol) was dissolved in dichloromethane (150 mL). At 0 °C, 85% metachloroperbenzoic acid (83.7 g, 412.1 mmol) was added portionwise. The mixture was gradually warmed to room temperature and stirred overnight. The reaction mixture was filtered, and the filter cake was dissolved in dichloromethane / methanol = 10:1 (800 mL) and washed with saturated sodium bicarbonate solution (300 mL × 4). The organic phase was dried and concentrated to give (9H-Fluorenyl-9-yl)methyl (4-cyano-1,1-dioxotetrahydro-2H-thiopyran-4-yl)carbamate A3d (38.1 g) in 28.2% yield. LC / MS: [M+H] + =396.2.

[0109] Step 4 A3d (38.1 g, 96.2 mmol) was dissolved in a 4N solution of hydrochloric acid in dioxane (300 mL), and concentrated hydrochloric acid (400 mL) was added and stirred at 90° C. for 30 h. The reaction mixture was concentrated to 200 mL, water (400 mL) was added, and the mixture was triturated to precipitate a solid. The solid obtained by filtration was triturated with ethyl acetate (200 mL), filtered, and dried to obtain intermediate A3 (12.77 g, purity: 97.80%). LC / MS: [M+H] + =416.2; 1 H NMR (400MHz, DMSO-d6) δ7.91(d,3H),7.72(d,2H),7.43(t,2H),7.35(td,2H),4.35(d,2H),4.25(t,1H),3.09(s,4H),2.46(s,2H),2.29(s,2H).

[0110] Intermediate B [ka]

[0111] Step 1 At room temperature, compound B2 (3.5 g, 16.25 mmol) and TEA (4.93 g, 48.72 mmol) were added to a solution of compound B1 (2.1 g, 16.13 mmol) in DCM (21 mL). Further, T3P (50%, 20.69 g, 32.51 mmol) was added under ice bath. The mixture was stirred at room temperature for 16 hours, then concentrated, ethyl acetate and water were added, and the aqueous phase was subsequently extracted twice with ethyl acetate. The combined organic phase was washed with 1M HCl, then saturated NaHCO3 solution, dried over Na2SO4, filtered, and the filtrate was concentrated to give compound B3 (5 g), with a yield of 94.6%. The crude product was directly used in the next step. LC / MS: [M+H] + -56=272.2.

[0112] Step 2 Compound B3 (5g, 15.27mmol) is dissolved in TFA / DCM (v:v=1:1, 50mL) solution and stirred at room temperature for 2 hours.The reaction solution is concentrated, then dissolved in an appropriate amount of DCM, and then concentrated again, and this is repeated three times.After DCM is dried and concentrated, compound B4 (2.7g) is obtained, with a yield of 65.2%.This crude product is directly used in the next step reaction. LC / MS: [M+H] + =272.2.

[0113] Step 3 To a solution of compound B4 (2.7 g, 9.95 mmol) in DMF (30 mL) was added compound B5 (4.47 g, 14.92 mmol) at room temperature. After cooling in an ice bath, HATU (4.54 g, 11.94 mmol) and DIEA (7.71 g, 59.69 mmol) were added to the solution. The mixture was stirred at 0 °C for 2 hours and then concentrated. The residue was dissolved in an appropriate amount of CH3CN and purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound B6 (1.5 g) in a 29.4% yield. LC / MS: [M] + =516.4.

[0114] Step 4 LiOH (244 mg, 5.8 mmol) was added to a solution of compound B6 (1.5 g, 2.9 mmol) in THF (15 mL) in an ice bath, and the mixture was stirred for 2 hours at 0° C. The reaction mixture was adjusted to neutral pH with 1 M HCl, then concentrated. The residue was dissolved in an appropriate amount of CH3CN and purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound B7 (1.3 g) in a yield of 89.6%. LC / MS: [M] + =502.4.

[0115] Step 5 Compound B8 (763 mg, 4.4 mmol) was added to a solution of compound B7 (1.1 g, 2.2 mmol) in CH3CN (10 mL) at room temperature. After cooling in an ice bath, HATU (1.0 g, 2.6 mmol) and DIEA (1.41 g, 10.9 mmol) were added. The mixture was stirred at 0 °C for 2 h, concentrated, and the residue was dissolved in an appropriate amount of CH3CN. The mixture was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound B9 (1.3 g) in 89.8% yield. LC / MS: [M] + =658.6.

[0116] Step 6 Compound B9 (1.3 g, 1.97 mmol) was dissolved in HCl / dioxane (4N, 100 mL) and stirred at room temperature for 2 hours. After concentrating the reaction solution, compound intermediate B (hydrochloride, 1.08 g) was obtained in a yield of 98.2%. LC / MS: [M] + =558.6, 1H NMR(400MHz,MeOD) δ3.94(s,2H),3.76(t,J=6.2Hz,2H),3.66(t,J=4.5Hz,8H),3.52(dd,J=12.9,6.7Hz,4H),3.27-3.07(m,10H),2.73(s,3H),2.52 (t,J=6.1Hz,2H),2.23(td,J=7.6,3.8Hz,4H),1.61(s,4H),1.50(d,J=6.6Hz,2H),1.33(d,J=9.9Hz,12H),0.91(t,J=6.7Hz,3H).

[0117] Intermediate C [ka] Intermediate C was synthesized with reference to the manufacturing method of WO 2021 / 127460.

[0118] Intermediate D [ka] Intermediate D was synthesized with reference to the production method of WO 2021 / 041770.

[0119] Intermediate L1 [ka]

[0120] Step 1 To a solution of compound L1-1 (0.9 g, 3.52 mmol) in CH3CN (25 mL) at 0 °C, DIEA (2.73 g, 21.1 mmol), HATU (1.61 g, 4.22 mmol), and N-Cbz-N-methylethylenediamine (hydrochloride, 1.29 g, 5.28 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CH3CN / 0.03% TFA / water, 0 to 100% gradient elution) to give compound L1-2 (TFA salt, 1.35 g) in 73% yield. LC / MS: [M] + =410.3.

[0121] Step 2 To a solution of compound L1-2 (1.2 g, 2.29 mmol) in THF (50 mL) was added 10% Pd / C (120 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CHCN / 0.03% TFA / water, 0-100% gradient elution) to give compound intermediate L1 (TFA salt, 760 mg) in 85% yield. LC / MS: [M] + =276.2. 1 H NMR(400MHz,MeOD) δ3.99-3.91(m,2H),3.77(t,J=6.2Hz,2H),3.70-3.62(m,4H),3.62-3.52(m, 4H),3.23(s,9H),3.10(t,J=5.6Hz,2H),2.70(s,3H),2.57(t,J=6.2Hz,2H).

[0122] Intermediate L2 [ka]

[0123] Step 1 To a solution of compound L2-1 (0.6 g, 2.86 mmol) in CH3CN (20 mL) at 0 °C, DIEA (2.22 g, 17.2 mmol), HATU (1.31 g, 3.43 mmol), and N-Cbz-N-methylethylenediamine (hydrochloride, 1.05 g, 4.29 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CH3CN / 0.03% TFA / water, 0 to 100% gradient elution) to give compound L2-2 (TFA salt, 1.17 g) in 86% yield. LC / MS: [M] + =364.3.

[0124] Step 2 To a solution of compound L2-2 (1.0 g, 2.29 mmol) in THF (50 mL) was added 10% Pd / C (100 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CHCN / 0.03% TFA / water, 0-100% gradient elution) to give compound intermediate L2 (TFA salt, 0.66 mg) in 92% yield. LC / MS: [M] + =230. 1 H NMR(400MHz,DMSO-d6) δ8.47-8.37(m,2H),3.35-3.21(m,4H),3.05(s,9H),2.79(t,J=6.0Hz,2H),2.53-2.4 5(m,3H),2.16-2.09(m,2H),1.72-1.62(m,2H),1.60-1.51(m,2H),1.29-1.21(m,2H).

[0125] Intermediate L3 [ka] [ka]

[0126] Step 1 To a solution of compound L3-1 (CAS: 1118767-16-0, 4.0 g, 4.73 mmol) in CH3CN (45 mL) at 0 °C, DIEA (3.67 g, 28.37 mmol), HATU (2.16 g, 5.67 mmol), and 2-amino-N-(2-(2-(3-methoxy-3-oxopropoxy)ethoxy)ethyl)-N,N-dimethylethane-1-ammonium hydrochloride (2.13 g, 7.09 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound L3-2 (TFA salt, 3.05 g) in 53.6% yield. LC / MS: [M]+ =1090.8.

[0127] Step 2 A solution of compound L3-2 (2.4 g, 1.99 mmol) in THF (24 mL) was added with aqueous LiOH (143 mg, 5.97 mmol, 12 mL) at 0 °C and reacted at this temperature for 1 h. The reaction mixture was concentrated to remove THF, and the residue was adjusted to pH 7 with dropwise addition of 1 N HCl. The mixture was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound L3-3 (TFA salt, 1.1 g) in 46.4% yield. LC / MS: [M] + =1076.7.

[0128] Step 3 To a solution of compound L3-3 (0.8 g, 0.67 mmol) in CH3CN (12 mL) at 0 °C, DIEA (521 mg, 4.03 mmol), HATU (306 mg, 0.81 mmol), and N-Cbz-N-methylethylenediamine (hydrochloride, 245 mg, 1.01 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound L3-4 (TFA salt, 870 mg) in 93.8% yield. LC / MS: [(M+H) / 2] + =632.9.

[0129] Step 4 To a solution of compound L3-4 (650 mg, 0.56 mmol) in THF (20 mL) was added 10% Pd / C (65 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CHCN / 0.03% TFA / water) to give compound intermediate L3 (TFA salt, 570 mg) in 97.1% yield. LC / MS: [M] +=1132.8.

[0130] Intermediate L4 [ka]

[0131] Step 1 To a solution of compound A9 (3.0 g, 3.67 mmol) in CH3CN (40 mL) at 0 °C, DIEA (2.84 g, 22.0 mmol), HATU (1.67 g, 4.40 mmol), and N-(2-aminoethyl)-6-methoxy-N,N-dimethyl-6-oxohexane-1-ammonium hydrochloride (1.4 g, 5.50 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound L4-1 (TFA salt, 1.75 g) in 42.2% yield. LC / MS: [M] + =1016.7.

[0132] Step 2 A solution of compound L4-1 (1.4 g, 1.24 mmol) in THF (15 mL) was added with aqueous LiOH (89 mg, 3.71 mmol, 5 mL) at 0 °C and reacted at this temperature for 1 h. The reaction mixture was concentrated to remove THF, and the residue was adjusted to pH 7 by dropwise addition of 1 N HCl. The mixture was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CHCN / 0.03% TFA / water) to give compound L4-2 (TFA salt, 740 mg) in 53.6% yield. LC / MS: [M] + =1002.7.

[0133] Step 3 To a solution of compound L3-3 (0.7 g, 0.63 mmol) in CH3CN (10 mL) at 0 °C, DIEA (486 mg, 3.76 mmol), HATU (286 mg, 0.75 mmol), and N-Cbz-N-methylethylenediamine (hydrochloride, 229 mg, 0.94 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CH3CN / 0.03% TFA / water) to give compound L4-3 (TFA salt, 770 mg) in 94.0% yield. LC / MS: [(M+H) / 2] + =596.9.

[0134] Step 4 To a solution of compound L4-3 (0.7 g, 0.56 mmol) in THF (20 mL) was added 10% Pd / C (70 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (5% to 40%, 40% to 70%, CHCN / 0.03% TFA / water) to give compound intermediate L4 (TFA salt, 590 mg) in 93.9% yield. LC / MS: [M] + =1044.8. 1 H NMR(400MHz,MeOD) δ4.31-4.23(m,1H),4.04(d,J=10.8Hz,4H),3.77-3.67(m,10H),3.64-3 .56(m,4H),3.53-3.45(m,6H),3.43-3.36(m,4H),3.20-3.12(m,8H),2. 75(s,3H),2.37-2.20(m,8H),2.18-2.09(m,1H),1.98-1.81(m,4H),1.7 9-1.70(m,2H),1.69-1.53(m,5H),1.47(d,J=8.0Hz,18H),1.31(s,20H).

[0135] Intermediate L5 [ka]

[0136] Step 1 To a solution of compound L3-1 (2 g, 2.36 mmol) in CHCN (20 mL) at 0 °C, DIEA (1.83 g, 14.2 mmol), HATU (1.08 g, 2.84 mmol), and N-Cbz-N-methylethylenediamine (hydrochloride, 864 mg, 3.55 mmol) were added sequentially, and the solution was stirred at 0 °C for 1 h. The mixture was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CHCN / 0.03% TFA / water, 0-100% gradient elution) to give compound L5-1 (1.9 g) in a 77.6% yield. LC / MS: [M+H] + =1036.7.

[0137] Step 2 To a solution of compound L1-2 (1.2 g, 1.16 mmol) in THF (30 mL) was added 10% Pd / C (120 mg) and the reaction was stirred under a hydrogen atmosphere (1.5 atm) at room temperature for 16 hours. The reaction mixture was filtered through diatomaceous earth, the filtrate was concentrated under reduced pressure, and the residue was purified by reverse-phase column chromatography (CHCN / 0.03% TFA / water, 0-100% gradient elution) to give compound L5 (940 mg) in 90% yield. LC / MS: [M+H] + =902.6. 1H NMR(400MHz,MeOD) δ4.27(dd,J=9.1,5.2Hz,1H),4.04(d,J=5.9Hz,4H),3.70(s,6H),3.68(d,J=5.6Hz,3H),3.62(dd,J=12. 1,6.8Hz,3H),3.56(dd,J=9.5,5.7Hz,4H),3.51-3.44(m,3H),3.41(d,J=5.9Hz,2H),3.08(t,J=5.8Hz,2 H),2.67(s,3H),2.31(t,J=7.6Hz,2H),2.28-2.19(m,5H),2.13(dd,J=13.7,5.9Hz,1H),2.05(d,J=5.5H z,1H),1.91(dd,J=13.9,9.1Hz,1H),1.61(dd,J=16.0,8.1Hz,4H),1.48(d,J=9.0Hz,18H),1.31(s,24H).

[0138] Example 1 [ka] [ka]

[0139] Step 1: Synthesis of fully protected linear peptide 1.1 Resin hook 1.1.1 Weigh 0.6 g of 2-CTC Resin (degree of substitution S = 1.08 mmol / g) and 78 mg of N-FMOC-L-3-pyridylalanine (0.2 mmol, degree of substitution 0.33) and add them to a reaction column. Add DCM (10 mL) and then 0.6 mL of DIEA to the reaction column and bubble with nitrogen gas for 2 hours. Add 0.6 mL of MeOH to the reaction column and continue to bubble with nitrogen gas for 30 minutes. Discharge the waste until no liquid flows out. Add DMF (30 mL) and wash five times, each time for 1 minute. Discharge the waste until no liquid flows out.

[0140] 1.1.2 20% piperidine / DMF (15 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was discharged until no liquid flowed out. DMF (30 mL) was added and washed five times, each time for 1 min. The waste was discharged until no liquid flowed out. The resin was detected with ninhydrin and was blue.

[0141] 1.2. Amino acid coupling 1.2.1 Coupling of Fmoc-Asn(Trt)-OH [Table 1]

[0142] 1. Weigh out 3.0 eq of Fmoc-Asn(Trt)-OH and add it to the resin. 6.00 eq of DIEA was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, 2.85 eq of HBTU was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0143] The reaction was carried out in an environment of 2.25°C for 0.5 hours, and the resin was detected with ninhydrin, and found to be colorless and transparent.

[0144] 3. The reaction solution was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0145] 1.2.2 Coupling of Fmoc-Glu(OtBu)-OH [Table 2]

[0146] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0147] 2. Weigh out 3.0 eq of Fmoc-Glu(OtBu)-OH and add it to the resin. Add 6.00 eq of DIEA, add 5 mL of DMF to the reaction column, and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to bubble evenly through the resin.

[0148] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0149] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0150] 1.2.3 Coupling of Fmoc-Thp(Gly)-OH [Table 3]

[0151] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0152] 2. Weigh out 3.0 eq of Fmoc-Thp(Gly)-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HATU. Adjust the nitrogen gas flow to bubble evenly through the resin.

[0153] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0154] 4. The reaction solution was drained and washed five times with DMF, each time for 1 minute, and waste was drained until no more liquid flowed out.

[0155] 1.2.4 Coupling of Fmoc-2Nal-OH [Table 4]

[0156] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was discharged until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was discharged until no more liquid flowed out. The resin was detected with tetrachlorobenzoquinone and was green.

[0157] 2. Weigh out 3.0 eq of Fmoc-2-Nal-OH and add it to the resin. 6.00 eq of DIEA was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, 2.85 eq of HATU was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0158] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with tetrachlorobenzoquinone, revealing that the resin was colorless and transparent.

[0159] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0160] 1.2.5 Coupling of Fmoc-4-[2-(Boc-amino)ethoxy]-L-Phenylalanine [Table 5]

[0161] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0162] 2. Weigh out 3.0 eq of Fmoc-4-[2-(Boc-amino)ethoxy]-L-phenylalanine and add it to the resin. 6.00 eq of DIEA was added. 5 mL of DMF was added to the reaction column, and nitrogen gas was bubbled through. After the amino acid had dissolved, 2.85 eq of HBTU was added. The nitrogen gas was adjusted to allow uniform bubbling throughout the resin.

[0163] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0164] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0165] 1.2.6 Coupling of Fmoc-Pen(Trt)-OH [Table 6]

[0166] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0167] 2. Weigh out 3.0 eq of Fmoc-Pen(Trt)-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to bubble evenly through the resin.

[0168] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0169] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0170] 1.2.7 Coupling of Fmoc-Lys(Ac)-OH [Table 7]

[0171] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0172] 2. Weigh out 3.0 eq of Fmoc-Lys(Ac)-OH and add it to the resin. Add 6.00 eq of DIEA, add 5 mL of DMF to the reaction column, and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to uniformly bubble through the resin.

[0173] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0174] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0175] 1.2.8 Coupling of Fmoc-(7Me)Trp-OH [Table 8]

[0176] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0177] 2. Weigh out 3.0 eq of Fmoc-(7Me)Trp-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to uniformly bubble through the resin.

[0178] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0179] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0180] 1.2.9 Coupling of Fmoc-Thr(Tbu)-OH [Table 9]

[0181] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0182] 2. Weigh out 3.0 eq of Fmoc-Thr(Tbu)-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to bubble evenly through the resin.

[0183] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0184] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0185] 1.2.10 Coupling of Fmoc-Asn(Trt)-OH [Table 10]

[0186] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0187] 2. Weigh out 3.0 eq of Fmoc-Asn(Trt)-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to uniformly bubble through the resin.

[0188] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0189] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0190] 1.2.11 Coupling of Fmoc-Pen(Trt)-OH [Table 11]

[0191] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0192] 2. Weigh out 3.0 eq of Fmoc-Pen(Trt)-OH and add it to the resin. Add 6.00 eq of DIEA. Add 5 mL of DMF to the reaction column and bubble with nitrogen gas. After the amino acid was dissolved, add 2.85 eq of HBTU. Adjust the nitrogen gas flow to bubble evenly through the resin.

[0193] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0194] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0195] 1.2.12 Ac2O end-capping [Table 12]

[0196] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 min. The waste was drained until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 min. The waste was drained until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0197] 2. DMF (10 mL), DIEA (6.00 eq), and Ac2O (5.0 eq) were added to the resin, and nitrogen gas was bubbled through the resin. The nitrogen gas was adjusted to bubble uniformly through the resin.

[0198] The reaction was carried out in an environment of 3.25°C for 10 minutes, and the resin was detected with ninhydrin, revealing that the resin was colorless and transparent.

[0199] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), 1 min each time, and waste was drained until no more liquid flowed out.

[0200] Step 2: Cleavage of the fully protected linear peptide 2.1 Wash five times with DMF (20 mL each time), 1 min each time, and drain the waste until no more liquid flows out.

[0201] 2.2 The resin was shrunk with MeOH (20 mL) for 3 min at a time, the waste was drained until no liquid flowed, and the resin was washed three times with 20 mL of methyl tert-butyl ether. The resin was dried with nitrogen gas and prepared for use.

[0202] 2.3 50 mL of the prepared 20% HFIP / DCM solution was placed in a reactor and bubbled with nitrogen gas. The reaction was carried out once every 30 minutes. After three cuts, all the filtrates were collected, concentrated under reduced pressure, and dried. An additional 50 mL of DCM was added, concentrated under reduced pressure, and continued to dry.

[0203] 2.4 100 mL of acetonitrile and 100 mL of water were added, and the mixture was sonicated until the product was uniformly dispersed. The mixture was then freeze-dried in a freeze dryer to obtain 500 mg of intermediate 1c, with a purity of 75% and a yield of 62%. LC / MS: [M+H] + =3012.7;[M+H-243] + =2769.7

[0204] Step 3: Condensation of Intermediate 1c with Intermediate A 200 mg (0.06 mmol) of the above intermediate 1c was taken, 1 mL of DMF was added, DIEA (3 eq, 0.18 mmol), and further intermediate A (77 mg, 0.06 mmol, 1 eq) was added, and HATU (23 mg, 0.06 mmol, 1 eq) was slowly added dropwise. The mixture was dissolved in DMF (0.2 mL). Tof-LCMS was used to detect the complete consumption of the raw material. The polarity of the product was very small, and the gradient was 80-100°C, and it took 2 min. LC / MS: [M] + =4099.20, [M-243] + =3856.2.

[0205] After the reaction was complete, the DMF was spun dry with an oil pump and prepared for use.

[0206] Step 4: Cleavage of the linear peptide 4.1. The cutting solution was mixed according to the following volumes:

[0207] [Table 13]

[0208] The crude peptide obtained in step 3 was added to the combined cleavage solution, shaken on a shaker for 2.5 hours, filtered, and the filtrate was added to 10 volumes of ice-cold isopropyl ether, centrifuged, and washed three times with isopropyl ether. The crude peptide was obtained after drying in vacuo for 2 hours. LC / MS: [(M+H) / 2] + =1402.8, [(M+2H) / 3] + =936.1.

[0209] Step 5: Oxidation of disulfide bonds 5.1 160 mg of the crude peptide obtained in Step 4 was dissolved in 100 mL of water and 100 mL of acetonitrile, and after clarification, 0.1 M I2 / MeOH was slowly added dropwise until the solution turned bright yellow. After stirring for 5 minutes, the yellow color did not disappear, and 0.1 M sodium thiosulfate was added dropwise until the yellow color disappeared. After stirring for 2 minutes, the yellow color remained unchanged. The sample was lyophilized and purified to obtain 43 mg of product Example 1, with a purity of 97.41% and a yield of 18.7%. LC / MS: [(M+H) / 2] + =1401.8;[(M+2H) / 3] + =935.1.

[0210] Purification conditions: [Table 14]

[0211] Example 2 [ka]

[0212] Referring to the synthesis method of Example 1, the reaction of intermediate B with polypeptide intermediate 1c and subsequent similar experimental procedures gave 26.4 mg of product Example 2, with a purity of 92.94% and a yield of 13.6%. LC / MS: [(M+H) / 2] + =1184.8;[(M+2H) / 3] + =790.2.

[0213] Example 3 [ka]

[0214] Referring to the synthesis method of Example 1, the reaction of intermediate L1 with polypeptide intermediate 1c and subsequent similar experimental procedures gave 42.0 mg of product Example 3, with a purity of 96.4% and a yield of 20.2%. LC / MS: [(M+H) / 2] + =1043.5.

[0215] Example 4 [ka]

[0216] Referring to the synthesis method of Example 1, the reaction of intermediate L3 with polypeptide intermediate 1c and subsequent similar experimental procedures gave 28.1 mg of product Example 4, with a purity of 96.2% and a yield of 14.9%. LC / MS: [(M+H) / 2] + =1415.7;[(M+2H) / 3] + =944.2.

[0217] Example 6 [ka]

[0218] Referring to the synthesis method of Example 1, the reaction of intermediate L5 with polypeptide intermediate 1c and subsequent similar experimental procedures gave 31.2 mg of product Example 6, with a purity of 97.8% and a yield of 18.1%. LC / MS: [(M+H) / 2] + =1300.1;[(M+2H) / 3] + =867.1.

[0219] Example 12 [ka]

[0220] Referring to the synthesis method of Example 1, the reaction of intermediate L2 with polypeptide intermediate 1c and subsequent similar experimental procedures gave 23.8 mg of product Example 12, with a purity of 95.7% and a yield of 17.5%. LC / MS: [(M+H) / 2] + =1020.5.

[0221] Example 12 can also be prepared by the following method. [ka]

[0222] Step 1: Condensation of Intermediate 1c with Intermediate L2 Intermediate 1c (200 mg, 0.06 mmol) from above was taken and added to DMF (1 mL), DIEA (10 eq, 0.6 mmol), and then Intermediate L2 (28 mg, 0.12 mmol, 2 eq). HATU (69 mg, 0.18 mmol, 3 eq) was slowly added dropwise. The mixture was dissolved in DMF (0.2 mL). Tof-LCMS was used to detect complete consumption of the starting material. After the reaction was complete, the DMF was spin-dried to give Intermediate 12d, which was ready for use.

[0223] Step 2: Deprotection The cutting solution was formulated according to the following volumes:

[0224] [Table 15]

[0225] The intermediate 12d obtained in the previous step was added to the compounded cleavage solution, shaken on a shaker for 2.5 hours, filtered, the filtrate was added to 10 volumes of ice-cold isopropyl ether, centrifuged, washed three times with isopropyl ether, and dried in vacuum for 2 hours to obtain intermediate 12e.

[0226] Step 3: Oxidation of disulfide bonds and purification, conversion to salt To the intermediate 12e (190 mg) obtained above, water (160 mL) and acetonitrile (40 mL) were added, and after dissolving and clarifying, 0.1 M I2 / MeOH was slowly added dropwise until the solution turned bright yellow. After stirring for 5 minutes, the yellow color did not disappear, and 0.1 M sodium thiosulfate was added dropwise until the yellow color disappeared. After stirring for 2 minutes, the reaction mixture of Example 12 was obtained.

[0227] 3.1 Purification The reaction mixture was purified under the following conditions.

[0228] Purification conditions: [Table 16]

[0229] After the purified solution was detected by RP-HPLC, the parts that passed the purity test were combined and diluted 2-fold with purified water for use.

[0230] 3.2 Conversion to Salts and Freeze-Drying Conversion conditions to salt: [Table 17]

[0231] The same preparative RP-HPLC column was equilibrated with 5% MPB in MPA at 15 ml / min for 15 minutes (MPA = 0.5% AcOH / water, MPB = 0.5% AcOH / ACN, MPC = 0.15M NHOAc / water). The diluted purified solution was loaded onto the chromatography column at 15 ml / min. It was washed with 5% MPB / MPC for 20 minutes, followed by 5% MPB / MPA for 20 minutes. Finally, the product was eluted with 60% MPB / MPA. The mixture was concentrated under reduced pressure to remove the organic solvent and lyophilized to give the final product, Example 12, weighing 35 mg, with a purity of 96.1% and a yield of 18.4%. LC / MS: [(M+H) / 2] + =1020.5.

[0232] Example 20 [ka]

[0233] Referring to the synthesis method of Example 1, the reaction of intermediate L4 with polypeptide intermediate 1c and subsequent similar experimental procedures gave 35.1 mg of product Example 20, with a purity of 97.1% and a yield of 19.1%. LC / MS: [(M+H) / 2] + =1378.7;[(M+2H) / 3] + =919.5.

[0234] Other examples were manufactured with reference to the manufacturing method described above.

[0235] [Table 18-1] [Table 18-2] [Table 18-3] [Table 18-4]

[0236] Examples 1-14 [ka]

[0237] Step 1: Fully protected linear peptide solid-phase synthesis 1.1 Resin hanger 1.1.1 Swelling Rink Amide-AM Resin (1-14a, 0.4 g, substitution degree 0.5-0.7 mmol / g) was weighed and added to a reaction column. DMF (10 mL) was added, and nitrogen gas was bubbled through the column for 20 minutes. The waste was then discharged until no more liquid flowed out.

[0238] 1.1.2 Resin hanger 1) 20% piperidine / DMF (10 mL) was added, and nitrogen gas was bubbled through for 30 minutes. The waste was evacuated until no more liquid flowed. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed. A sample was taken and detected with ninhydrin, and the resin was blue.

[0239] 2) Fluorenylmethoxycarbonylsarcosine (186.6 mg, 0.6 mmol), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0240] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through for 4 hours.

[0241] 4) The solvent was removed. The resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was continuously bubbled through the resin, and the resin was stirred for 2-3 minutes, after which the solvent was removed. A sample was taken and detected with ninhydrin, and the resin was found to be colorless.

[0242] 1.2. Amino acid coupling 1.2.1 Coupling of Fmoc-3-(4-pyridyl)-L-alanine [Table 19]

[0243] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with tetrachlorobenzoquinone, and the resin was blue in color.

[0244] 2) Fmoc-3-(4-pyridyl)-L-alanine (234 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0245] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. Sampling was performed and detection with tetrachlorobenzoquinone revealed that the resin was colorless and transparent.

[0246] 4) The solvent was removed, and the resin was washed sequentially with 10 mL of DMF (10 mL), 10 mL of isopropanol (10 mL), 10 mL of DMF (10 mL), 10 mL of DMF (10 mL), and 10 mL of DMF (10 mL). After each wash, nitrogen gas was maintained under stirring for 2-3 minutes, and the solvent was then removed.

[0247] 1.2.2 Coupling of Fmoc-Asn(Trt)-OH [Table 20]

[0248] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0249] 2) Fmoc-Asn(Trt)-OH (358 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0250] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0251] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0252] 1.2.3 Coupling of Fmoc-Glu(OtBu)-OH [Table 21]

[0253] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0254] 2) Fmoc-Glu(OtBu)-OH (255 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0255] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0256] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0257] 1.2.4 Coupling of 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexaneformic acid [Table 22]

[0258] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0259] 2) 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexaneformic acid (241 mg), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0260] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0261] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0262] 1.2.5 Coupling of Fmoc-2Nal-OH [Table 23]

[0263] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with tetrachlorobenzoquinone, revealing that the resin was reddish brown.

[0264] 2) Fmoc-2Nal-OH (262 mg), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0265] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. Sampling was performed and detection with tetrachlorobenzoquinone revealed that the resin was colorless and transparent.

[0266] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0267] 1.2.6 Coupling of Fmoc-4-[2-(Boc-amino)ethoxy]-L-Phenylalanine [Table 24]

[0268] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0269] 2) Fmoc-4-[2-(Boc-amino)ethoxy]-L-phenylalanine (328 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0270] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0271] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0272] 1.2.7 Coupling of Fmoc-Pen(Trt)-OH [Table 25]

[0273] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0274] 2) Fmoc-Pen(Trt)-OH (368 mg), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0275] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0276] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0277] 1.2.8 Coupling of Fmoc-Lys(Ac)-OH [Table 26]

[0278] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0279] 2) Fmoc-Lys(Ac)-OH (246 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0280] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0281] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0282] 1.2.9 Coupling of Fmoc-(7Me)Trp-OH [Table 27]

[0283] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0284] 2) Fmoc-(7Me)Trp-OH (264 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0285] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0286] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0287] 1.2.10 Coupling of Fmoc-Thr(Tbu)-OH [Table 28]

[0288] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0289] 2) Fmoc-Thr(Tbu)-OH (238 mg), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0290] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0291] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0292] 1.2.11 Coupling of Fmoc-Asn(Trt)-OH [Table 29]

[0293] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0294] 2) Fmoc-Asn(Trt)-OH (358 mg), N,N'-diisopropylcarbodiimide (76 mg), and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0295] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0296] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0297] 1.2.12 Coupling of Fmoc-Pen(Trt)-OH [Table 30]

[0298] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0299] 2) Fmoc-Pen(Trt)-OH (368 mg), N,N'-diisopropylcarbodiimide (76 mg) and 1-hydroxybenzotriazole (81 mg) were weighed and added to a reaction column.

[0300] 3) DMF (10 mL) was added and dissolved, and nitrogen gas was bubbled through, followed by reaction at room temperature for 3 hours. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0301] 4) The solvent was removed, and the resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was maintained under bubbling, and the resin was stirred for 2-3 minutes, after which the solvent was removed again.

[0302] 1.2.13 Ac2O end-capping [Table 31]

[0303] 1) 20% piperidine / DMF (10 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was evacuated until no more liquid flowed out. DMF (10 mL) was added and washed five times, each time for 1 minute. The waste was evacuated until no more liquid flowed out. A sample was taken and detected with ninhydrin, and the resin was blue.

[0304] 2) DCM (10 mL), DIEA (6.0 eq), and Ac2O (5.0 eq) were added to the resin, and nitrogen gas was bubbled through the resin. The nitrogen gas was adjusted to bubble uniformly through the resin.

[0305] 3) The reaction was carried out at room temperature for 30 minutes. A sample was taken and detected with ninhydrin, and the resin was found to be colorless and transparent.

[0306] 4) The resin was washed sequentially with DMF (10 mL), isopropanol (10 mL), DMF (10 mL), DMF (10 mL), and DMF (10 mL). After each wash, nitrogen gas was continuously bubbled through the resin, and the resin was stirred for 2-3 minutes, after which the solvent was removed.

[0307] 5) Sample collection and washing: The resin was washed with isopropyl ether (10 mL), bubbled with nitrogen gas, stirred, washed for 2-3 minutes, and dried by suction. This procedure was repeated three times to obtain resin peptide 1-14b.

[0308] Step 2: Cleavage and deprotection of the linear peptide The cutting solution was formulated according to the following volumes:

[0309] [Table 32]

[0310] The resulting resin peptide 1-14b was added to 10 mL of the cleavage solution, shaken in a water bath at 25°C for 2.5 hours, filtered, and the filtrate was added to 10 volumes of ice-cold isopropyl ether, centrifuged, and the supernatant was discarded. The resulting mixture was washed three times with isopropyl ether and vacuum-dried for 2 hours to obtain crude peptide 1-14c.

[0311] Step 3: Oxidation of disulfide bonds The crude peptide 1-14c (190 mg) obtained above was added to water (100 mL) and acetonitrile (100 mL), dissolved, and clarified. 0.1 M I2 / MeOH was slowly added dropwise until the solution turned bright yellow. After stirring for 5 minutes, the yellow color did not disappear. 0.1 M sodium thiosulfate was added dropwise until the yellow color disappeared. After stirring for 2 minutes, the sample was purified under the following conditions.

[0312] Purification conditions: [Table 33]

[0313] The purified solution was concentrated under reduced pressure to remove the organic solvent and freeze-dried to give 45 mg of the final product Example 1-14, with a purity of 97.5% and a yield of 11.7%. MS: m / z=1931.8, [M+H] + , m / z=966.4, [M+2H] 2+ .

[0314] Example 1-16-1 [ka] [ka]

[0315] Step 1: Synthesis of fully protected linear peptide 1.1 Resin hanger 1.1.1 0.6 g of 2-CTC Resin (1-16-1a, degree of substitution S=1.08 mmol / g) and 78 mg of N-Fmoc-L-3-pyridylalanine (0.2 mmol, degree of substitution 0.33) were weighed and added to a reaction column. DCM (10 mL) was then added, followed by 0.6 mL of DIEA, and nitrogen gas was bubbled through the column for 2 hours. 0.6 mL of MeOH was then added to the reaction column, nitrogen gas was bubbled through the column for 30 minutes, and the waste was discharged until no more liquid flowed out. DMF (30 mL) was added and washed five times, each time for 1 minute, and the waste was discharged until no more liquid flowed out.

[0316] 1.1.2 20% piperidine / DMF (15 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was discharged until no more liquid flowed out. DMF (30 mL) was added and washed five times, each time for 1 minute. The waste was discharged until no more liquid flowed out. The resin was detected with ninhydrin and turned blue.

[0317] 1.2. Amino acid coupling 1.2.1 Coupling of Fmoc-Asn(Trt)-OH [Table 34]

[0318] 1. Fmoc-Asn(Trt)-OH (358 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0319] The reaction was carried out in an environment of 2.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0320] 3. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0321] 1.2.2 Coupling of Fmoc-Glu(OtBu)-OH [Table 35]

[0322] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed. The resin was detected with ninhydrin and was blue.

[0323] 2. Fmoc-Glu(OtBu)-OH (255 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column, and nitrogen gas was bubbled through. After the amino acid was dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0324] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0325] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0326] 1.2.3 Coupling of 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexaneformic acid [Table 36]

[0327] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0328] 2.1-(9H-Fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluorocyclohexaneformic acid (CAS: 1986905-26-3, 241 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added, and 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, HATU (217 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0329] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0330] 4. The reaction mixture was drained and washed five times with DMF, 1 minute each time, and waste was drained until no more liquid flowed.

[0331] 1.2.4 Coupling of Fmoc-2Nal-OH [Table 37]

[0332] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no liquid flowed out. The resin was green when detected with tetrachlorobenzoquinone.

[0333] 2. Fmoc-2-Nal-OH (262 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid dissolved, HATU (217 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0334] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with tetrachlorobenzoquinone, and was found to be colorless and transparent.

[0335] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0336] 1.2.5 Coupling of Fmoc-4-[2-(Boc-amino)ethoxy]-L-Phenylalanine [Table 38]

[0337] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0338] 2. Fmoc-4-[2-(Boc-amino)ethoxy]-L-phenylalanine (328 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column, and nitrogen gas was bubbled through. After the amino acid had dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0339] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0340] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0341] 1.2.6 Coupling of Fmoc-Pen(Trt)-OH [Table 39]

[0342] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0343] 2. Fmoc-Pen(Trt)-OH (368 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0344] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0345] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0346] 1.2.7 Coupling of Fmoc-Lys(Ac)-OH [Table 40]

[0347] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0348] 2. Fmoc-Lys(Ac)-OH (246 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0349] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0350] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0351] 1.2.8 Coupling of Fmoc-(7Me)Trp-OH [Table 41]

[0352] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0353] 2. Fmoc-(7Me)Trp-OH (264 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid dissolved, HATU (217 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0354] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0355] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0356] 1.2.9 Coupling of Fmoc-Thr(Tbu)-OH [Table 42]

[0357] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0358] 2. Fmoc-Thr(Tbu)-OH (238 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0359] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0360] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0361] 1.2.10 Coupling of Fmoc-Asn(Trt)-OH [Table 43]

[0362] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0363] 2. Fmoc-Asn(Trt)-OH (358 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0364] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0365] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0366] 1.2.11 Coupling of Fmoc-Pen(Trt)-OH [Table 44]

[0367] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0368] 2. Fmoc-Pen(Trt)-OH (368 mg, 3.0 eq) was weighed and added to the resin. DIEA (155 mg, 6.00 eq) was added. 5 mL of DMF was added to the reaction column. Nitrogen gas was bubbled through the column. After the amino acid was dissolved, HBTU (216 mg, 2.85 eq) was added. The nitrogen gas was adjusted to bubble evenly through the resin.

[0369] The reaction was carried out in an environment of 3.25°C for 0.5 hours, and the resin was detected with ninhydrin, and was found to be colorless and transparent.

[0370] 4. The reaction mixture was drained and washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed.

[0371] 1.2.12 Ac2O end-capping [Table 45]

[0372] 1.20% piperidine / DMF (20 mL) was added to the reaction column, and nitrogen gas was bubbled through for 20 minutes. The waste was emptied until no more liquid flowed out. DMF (20 mL) was added and washed five times, each time for 1 minute. The waste was emptied until no more liquid flowed out. The resin was detected with ninhydrin and was blue.

[0373] 2. DMF (10 mL), DIEA (155 mg, 6.00 eq), and Ac2O (102 mg, 5.0 eq) were added to the resin, and nitrogen gas was bubbled through the resin. The nitrogen gas was adjusted to bubble uniformly through the resin.

[0374] The reaction was carried out in an environment of 3.25°C for 10 minutes, and detection with ninhydrin revealed that the resin was colorless and transparent.

[0375] 4. The reaction solution was drained, and the mixture was washed five times with DMF (20 mL each time), for 1 minute each time, and waste was drained until no more liquid flowed out, to obtain linear peptide 1-16-1b.

[0376] Step 2: Cleavage of the fully protected linear peptide 2.1 Wash five times with DMF (20 mL each), 1 minute each time, and drain the waste until no more liquid flows.

[0377] 2.2 The resin was shrunk with MeOH (20 mL) for 3 min at a time, drained until no liquid flowed, and washed with 20 mL of methyl tert-butyl ether three times. The resin was dried with nitrogen gas and prepared for use.

[0378] 2.3 50 mL of the prepared 20% HFIP / DCM solution was placed in a reactor and bubbled with nitrogen gas. The reaction was carried out once every 30 minutes. After three cuts, all the filtrates were collected, concentrated under reduced pressure, and dried. An additional 50 mL of DCM was added, concentrated under reduced pressure, and continued to dry.

[0379] 2.4 100 mL of acetonitrile and 100 mL of water were added, and the mixture was sonicated until the product was uniformly dispersed. The mixture was then freeze-dried in a freeze dryer to obtain 480 mg of intermediate 1-16-1c, with a purity of 76% and a yield of 60%. LC / MS: [M+H] + =3044.4, [M+H-243] + =2801.4.

[0380] Step 3: Condensation of Intermediate 1-16-1c with Intermediate A The above intermediate 1-16-1c (200 mg, 0.06 mmol) was taken, and 1 mL of DMF and DIEA (3 eq, 0.18 mmol) were added, followed by the addition of Intermediate A (77 mg, 0.06 mmol, 1 eq), and then HATU (23 mg, 0.06 mmol, 1 eq, dissolved in DMF (0.2 mL)) was slowly added dropwise. Tof-LCMS was used to detect the complete consumption of the raw material, and the DMF was spun dry on an oil pump to give crude intermediate 1-16-1d, which was ready for use. LC / MS: [M] + =4130.2,[M-243] + =3887.2.

[0381] Step 4: Cleavage of the linear peptide 4.1. The cutting solution was mixed according to the following volumes:

[0382] [Table 46]

[0383] The crude peptide obtained in step 3 was added to the cleavage solution, shaken on a shaker for 2.5 h, filtered, and the filtrate was added to 10 volumes of ice-cold isopropyl ether, centrifuged, washed three times with isopropyl ether, and dried in vacuo for 2 h to obtain crude peptide 1-16-1e. LC / MS: [(M+H) / 2] + =1419.2, [(M+2H) / 3] + =946.5.

[0384] Step 5: Oxidation of disulfide bonds 5.1 160 mg of crude peptide 1-16-1e obtained in Step 4 was dissolved in 100 mL of water and 100 mL of acetonitrile, and after clarification, 0.1 M I2 / MeOH was slowly added dropwise until the solution turned bright yellow. After stirring for 5 minutes, the yellow color did not disappear, and 0.1 M sodium thiosulfate was added dropwise until the yellow color disappeared. After stirring for 2 minutes, the yellow color remained unchanged. The sample was lyophilized and purified to obtain 42 mg of product Example 1-16-1, with a purity of 97.1% and a yield of 18.0%. LC / MS: [(M+H) / 2] + =1418.2, [(M+2H) / 3] + =945.8.

[0385] Purification conditions: [Table 47]

[0386] Example 3-16-1 [ka]

[0387] Example 3-16-1 can be produced by referring to the following method. Refer to the preparation method of Example 1-16-1, in which 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexaneformic acid in step 1 of linear peptide solid phase synthesis is replaced with (9H-fluorenyl-9-yl)methyl (4-cyanotetrahydro-2H-thiopyran-4-yl)carbamate A3c to obtain 120 mg of product Example 3-16-1, with a purity of 98.5% and a yield of 23.0%. LC / MS: [(M+H) / 2] + =1409.6, [(M+2H) / 3] + =940.8.

[0388] Example 4-16-1 [ka]

[0389] Example 4-16-1 can be produced with reference to the following method. Refer to the preparation method of Example 1-16-1, in which 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexane formic acid in step 1 of linear peptide solid phase synthesis is replaced with intermediate A3, to obtain 110 mg of product Example 4-16-1, with a purity of 99.4% and a yield of 21.0%. LC / MS: [(M+H) / 2] + =1425.7, [(M+2H) / 3] + =951.0.

[0390] Purification conditions: [Table 48]

[0391] Examples 1-19 [ka]

[0392] Examples 1-19 can be prepared with reference to the following methods. Refer to the preparation method of Example 1-16-1, where intermediate A in the coupling of the linear peptide and the side chain in step 3 was replaced with intermediate L2, and 110 mg of product Example 1-19 was obtained, with a purity of 95.1% and a yield of 15.4%. LC / MS: [(M+H) / 2] + =1037.5, [(M+2H) / 3] + =692.4.

[0393] Example 3-19 [ka]

[0394] Examples 3-19 can be prepared with reference to the following methods. Refer to the preparation method of Example 1-16-1, in which 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexaneformic acid in step 1 of the linear peptide solid phase synthesis is replaced with (9H-fluorenyl-9-yl)methyl(4-cyanotetrahydro-2H-thiopyran-4-yl)carbamate A3c, and in step 3 of the linear peptide and side chain bonding, intermediate A, which is the side chain, is replaced with intermediate L2, to obtain 110 mg of product Example 3-19, with a purity of 99.0% and a yield of 15.4%. LC / MS: [(M+H) / 2] + =1028.5, [(M+2H) / 3] + =686.4.

[0395] Example 4-19 [ka]

[0396] Examples 4-19 can be prepared with reference to the following methods. Refer to the preparation method of Example 1-16-1, in which 1-(9H-fluorenyl-9-ylmethoxycarbonylamino)-4,4-difluoro-cyclohexane formic acid in step 1 of the linear peptide solid phase synthesis is replaced with intermediate A3, and intermediate A, the side chain in the bonding between the linear peptide and the side chain in step 3, is replaced with intermediate L2, to obtain 102 mg of product Example 4-19, with a purity of 99.8% and a yield of 14.2%. LC / MS: [(M+H) / 2] + =1044.5, [(M+2H) / 3] + =697.0.

[0397] The following examples can be produced by referring to the above production methods. [ka] [ka] [ka] [ka] [ka] [ka]

[0398] where R 1 and R 2 is selected from the following structures:

[0399] [Table 49-1] [Table 49-2] [Table 49-3] [Table 49-4] [Table 49-5] [Table 49-6] [Table 49-7] [Table 49-8] [Table 49-9] [Table 49-10] [Table 49-11] [Table 49-12] [Table 49-13]

[0400] Biological Test Evaluation The present invention will be further explained below in conjunction with test examples, but these examples are not intended to limit the scope of the present invention.

[0401] Test Example 1: Experiment in which the compound of the present invention competes with IL23 for binding to IL23R 1.1 Experimental Objective: To determine the efficiency with which the compounds of the present invention compete with IL23 for binding to IL23R

[0402] 1.2 Laboratory equipment and reagents: 1.2.1 Equipment [Table 50]

[0403] 1.2.2 Reagents [Table 51]

[0404] 1.3 Experimental Method: The compounds of the present invention were screened using IL23 / IL23R BINDING ASSAY KITS (PerkinElmer Cat#64BDPIL23PEH). Screening of the compounds of the present invention was carried out in the form of a multiwell plate (suitable for HTRF assay). Generally, 2 μl of some of the compounds of the present invention and the standard solutions in the kit were first added to each well of the multiwell plate, and then 4 μl of Tag1-IL23, 4 μl of Tag2-IL23R, and 10 μL of premixed Anti-Tag1 Eu Cryptate Antibody and Anti-Tag2 d2 reagent were added to all wells. The multiwell plate was shielded from light using a non-transparent coating, and after incubation at room temperature for 2 hours, the non-transparent coating was removed. The signal value was read using an HTRF-compatible detection instrument to determine the IC. 50 The value was detected.

[0405] 1.4 Experimental data processing method The calculation of the inhibition rate was as follows: % inhibition=(Signal cmpd-Signal Ave_PC) / (Signal Ave_VC-Signal Ave_PC)×100. H=Ave(DMSO), L=Ave(Guselkumab)

[0406] Compound IC 50 values ​​were calculated. Nonlinear regression fitting was performed on the HTRF experimental data using Graphpad's log(inhibitor) vs. response - variable slope (four parameters) to fit the curve and calculate the IC. 50 got the value. Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope)) X:log of inhibitor concentration;Y:% Inhibition

[0407] 1.5 Experimental Results: [Table 52]

[0408] 1.6 Experimental Conclusion: The compounds of the preferred examples of the present invention showed excellent biological activity in the inhibition test of competition with IL23 for binding to IL23R.

[0409] Test Example 2: Experiment to test whether the compound of the present invention competes with IL12 for binding to IL12Rβ1 2.1 Experimental Objective: Efficiency of the Compounds of the Invention to Compete with IL12 for Binding to IL12Rβ1

[0410] 2.2 Laboratory equipment and reagents 2.2.1 Equipment [Table 53]

[0411] 2.2.2 Reagents [Table 54]

[0412] 2.3 Experimental Method: The compounds of the present invention were screened using IL12 / IL12RB1 BINDING ASSAY KITS (PerkinElmer Cat# 64BDIL12PEG). Screening of the compounds of the present invention was carried out in the form of a multiwell plate (suitable for HTRF assay). Generally, 2 μl of some compounds of the present invention and the standard solutions in the kit were first added to each well of the multiwell plate, and then 4 μl of Tag1-IL12, 4 μl of Tag2-IL12Rb1, and 10 μL of premixed Anti-Tag1 Eu Cryptate Antibody and Anti-Tag2 XL665 reagent were added to all wells. The multiwell plate was shielded from light using a non-transparent film, and after incubation at room temperature for 2 hours, the non-transparent film was removed. The signal value was read using an HTRF-compatible detection instrument to determine the IC. 50 The value was detected.

[0413] 2.4 Experimental data processing method The calculation of the inhibition rate was as follows: % inhibition =(Signal cmpd-Signal Ave_PC) / (Signal Ave_VC-Signal Ave_PC)×100. H=Ave(DMSO);L=Ave(Guselkumab)

[0414] Compound IC 50 values ​​were calculated. Nonlinear regression fitting was performed on the HTRF experimental data using Graphpad's log(inhibitor) vs. response - variable slope (four parameters) to fit the curve and calculate the IC. 50 got the value. Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope)) X:log of inhibitor concentration;Y:% Inhibition

[0415] 2.5 Experimental conclusions: According to the above method, the compounds of the present invention exhibited an IC value of 1000 nM (IC 50 ) showed greater biological activity than

[0416] Test Example 3: Experiment to block STAT3 phosphorylation in hIL23-activated DB cells by the compounds of the present invention 3.1 Experimental Objective: Measurement of the Inhibitory Activity of the Compounds of the Present Invention Against hIL23-Activated STAT3 Phosphorylation in DB Cells

[0417] 3.2 Laboratory equipment and reagents 3.2.1 Equipment [Table 55]

[0418] 3.2.2 Reagents [Table 56]

[0419] 3.3 Experimental Method: The compounds produced above were added to DB (ATCC Cat#CRL-2289) cells to inhibit the signal transduction of rhIL-23 cytokine. The screening of compounds was carried out in a multi-well plate format (suitable for ELISA assay), and generally, in the presence of some of the compounds of the present invention, rhIL-23 (0.5 nM, R&D SYSTEMS) was used at 6.25 x 10 6 DB cells (cells / milliliter) were stimulated in RPMI medium (Invitrogen) containing 10% FBS, and after half an hour, they were cleaved on ice with 50 μL of 2× cleavage solution (CST). The p-STAT3 content of the cleaved product was measured using the PathScan Phospho-Stat3 (Tyr705) Sandwich ELISA Kit (CST Cat#7300C), and the activity of the compounds of the present invention in blocking rhIL23 signaling in DB cells was calculated.

[0420] 3.4 Experimental data processing method The calculation of the inhibition rate was as follows: Inhibition%=(Ave_H-Sample) / (Ave_H-Ave_L)×100 H=Ave(DMSO), L=Ave(Guselkumab)

[0421] Compound IC 50 values ​​were calculated. Nonlinear regression fitting was performed on the ELISA experimental data using Graphpad's log(inhibitor) vs. response - variable slope (four parameters) to fit the curve and calculate the IC 50 got the value. Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)×HillSlope)) X:log of inhibitor concentration;Y:% Inhibition

[0422] 3.5 Experimental results: [Table 57]

[0423] 3.6 Experimental Conclusion: The compounds of the preferred examples of the present invention showed excellent inhibitory activity in the test for inhibiting hIL23-activated STAT3 phosphorylation.

[0424] Test Example 4: Pharmacokinetics measurement in rats 4.1. Research Objective: SD rats were used as test animals to investigate the pharmacokinetic behavior of the compounds of the examples in the rat body (plasma) when administered orally or intravenously.

[0425] 4.2. Testing method 4.2.1 Test Drugs: The compounds embodying the present invention were prepared in-house.

[0426] 4.2.2 Test animals: Male SD rats, three per group, were used. Shanghai Jie Sijie Laboratory Animal Co., Ltd., Animal Production Permit Number (SCXK (Shanghai) 2013-0006 No. 0.311620400001794).

[0427] 4.2.3 Drug Preparation: Preparation of oral medication: 20% Labrasol 20% Labrasol: 40 ml of Labrasol was weighed and placed in a 500 ml glass bottle, and PBS was added up to 200 ml. The solution was stirred magnetically and completely dissolved to prepare 20% Labrasol.

[0428] The example compound was weighed and dissolved in the solution, shaken well and sonicated for 5 minutes to obtain a clear, colorless solution with a concentration of 0.5 mg / mL.

[0429] Preparation of intravenous drugs: PBS The example compound was weighed and dissolved in the solution, vortexed, sonicated for 5 minutes, and passed through a 0.22 μm filter membrane to obtain a clear, colorless solution with a concentration of 0.2 mg / mL.

[0430] 4.2.4 Administration: Male SD rats (3 rats per group) were used. After an overnight fast, the drug was administered PO at a dose of 5 mg / kg in a volume of 10 mL / kg.

[0431] Male SD rats (3 rats per group) were used. After overnight fasting, the drug was administered intravenously at a dose of 1 mg / kg in a volume of 5 mL / kg.

[0432] 4.2.5 Sample Collection: 0.2 mL of blood was collected from the jugular vein before administration and at 0.083 h (IV), 0.25 h, 0.5 h, 1.0 h, 2.0 h, 4.0 h, 6.0 h, 8.0 h, and 24.0 h after administration. The blood was placed in an EDTA-2K test tube and centrifuged at 4°C, 6000 rpm for 6 minutes to separate the plasma. The plasma was stored at -20°C and served 4 h after administration.

[0433] 4.3 Sample Processing: 1) 50 μL of plasma sample was added to 200 μL of acetonitrile to precipitate, mixed, and then centrifuged at 4500 rpm for 15 minutes.

[0434] 2) The treated supernatant solution was collected and analyzed for the concentration of the test compound by LC / MS / MS, and the LC / MS / MS analysis equipment was AB Sciex Triple Quad 6500+.

[0435] 4.4 Liquid phase analysis: Liquid phase conditions: Shimadzu LC-30AD pump Chromatography column: HALO 90A PFP 2.7 μm 2.1×30 mm Mobile phase: Solution A was a 5% aqueous acetonitrile solution (0.1% formic acid), and solution B was a 95% aqueous acetonitrile solution (0.1% formic acid). ●Flow rate: 0.6mL / min Elution time: 0 to 2.0 minutes, and the eluate was as follows:

[0436] [Table 58]

[0437] 4.5. Test Results and Analysis The main pharmacokinetic parameters were calculated using WinNonlin 8.1.

[0438] [Table 59]

[0439] 4.6 Experimental conclusions: Data from pharmacokinetic evaluation experiments in rats showed that compounds of preferred embodiments of the present invention had stronger blood concentrations and higher exposures compared to PN-235.

[0440] Test Example 5: Pharmacokinetics measurement in beagle dogs 5.1. Research Objective: Beagle dogs were used as test animals to investigate the pharmacokinetic behavior of the compounds of the examples in the rat body (plasma) when administered IV.

[0441] 5.2. Testing method 5.2.1 Test Drugs: The compounds embodying the present invention were prepared in-house.

[0442] 5.2.2 Test animals: Male beagle dogs, 3 per group, were used. Yizheng Anlimao Biotechnology Co., Ltd. 5.2.3 Drug Preparation: Preparation of intravenous drugs: PBS The example compound was weighed and dissolved in the solution, vortexed, sonicated for 5 minutes, and passed through a 0.22 μm filter membrane to obtain a clear, colorless solution with a concentration of 0.1 mg / mL.

[0443] 5.2.4 Administration: Three male beagle dogs per group were administered IV after an overnight fast at a dose of 0.2 mg / kg in a volume of 2 mL / kg.

[0444] 5.2.5 Sample Collection: Before administration and at 0.083 h (IV), 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 24 h, 48 h, 72 h, 96 h, 120 h, 144 h, and 168 h after administration, 0.5 mL of blood was collected from the forelimb vein, placed in an EDTA-2K test tube, and centrifuged at 4°C, 6000 rpm for 6 minutes to separate the plasma, which was stored at -20°C and served 4 h after administration.

[0445] 5.3 Sample Processing: 1) 50 μL of plasma sample was added to 200 μL of acetonitrile to precipitate, mixed, and then centrifuged at 4500 rpm for 15 minutes.

[0446] 2) The treated supernatant solution was taken and analyzed for the concentration of the test compound by LC / MS / MS, and the LC / MS / MS analysis equipment was AB Sciex Triple Quad 6500+.

[0447] 5.4 Liquid phase analysis: ●Liquid phase conditions: ExionLC Chromatography column: ACQUITY™ Premier Peptide CSH C18 130A 1.7 μm 2.1 × 100 mm Column Mobile phase: Solution A was a 5% aqueous acetonitrile solution (0.1% formic acid), and solution B was a 95% aqueous acetonitrile solution (0.1% formic acid). ●Flow rate: 0.6mL / min Elution time: 0 to 2.0 minutes, and the eluate was as follows:

[0448] [Table 60]

[0449] 5.5. Test Results and Analysis The main pharmacokinetic parameters were calculated using WinNonlin 8.1.

[0450] [Table 61]

[0451] 5.6 Experimental conclusions: Data from pharmacokinetic evaluation studies in dogs showed that compounds of preferred embodiments of the present invention had stronger blood concentrations and higher exposures compared to PN-235.

[0452] Test Example 6: Measurement of the solubility of the compound of the present invention 6.1 Research objective: The solubility of the compounds in PBS was investigated.

[0453] 6.2 Laboratory equipment and reagents: [Table 62]

[0454] 6.3 Experimental Method: Approximately 2 mg of the compound was weighed out, added to an appropriate amount of PBS solution, and dissolved by ultrasonic treatment for 10 seconds. No precipitation was observed even after leaving it at room temperature overnight.

[0455] 6.4 Experimental results and conclusions: At room temperature, the compounds of the preferred examples of the present invention exhibited excellent solubility advantages, for example, the solubility of Example 1-16-1 was 39.58 to 59.38 mg / mL, which was greater than the solubility of Example 4-16-1, 124.75 mg / mL.

[0456] Test Example 7: Stability of the compound of the present invention in simulated gastrointestinal fluid 7.1 Purpose of the study: The stability of the compounds of the examples in simulated gastrointestinal fluid was investigated.

[0457] 7.2 Laboratory equipment and reagents: 7.2.1 Reagents Acetonitrile (F22M4L201, Fisher), trifluoroacetic acid (17110655, TEDIA), sodium hydroxide (20170616, China National Pharmaceutical Co., Ltd.), pancreatin (20220811, Shanghai Experimental Laboratory), pepsin (20220909, Shanghai Experimental Laboratory), potassium dihydrogen phosphate (20210528, Wokai), sodium chloride (20200105, Shanghai Experimental Laboratory), hydrochloric acid (20181203, China National Pharmaceutical Co., Ltd.), anhydrous sodium dihydrogen phosphate (20200602, Shanghai Experimental Laboratory), 3F Powder (FFF-0723-B, Biorelevant), glacial acetic acid (20181112, China National Pharmaceutical Co., Ltd.).

[0458] 7.2.2 Equipment High-performance liquid chromatography (1260, Agilent), electronic balance (MCE-C, Sartorius), pH meter (Five Easy Plus, METTLER TOLEDO), pipette (500-5000 μL, 100-1000 μL, 10-100 μL, Eppendorf), ultrasonic cleaner (SK5200LHC, Shanghai Ketuo Ultrasound Instrument Co., Ltd.).

[0459] 7.3 Experimental Method: 7.3.1 Preparation of mobile phase and GI simulant 0.05% trifluoroacetic acid aqueous solution: 2 L of purified water was measured out, 1 mL of trifluoroacetic acid was added, and the mixture was mixed uniformly and then ultrasonicated to obtain the solution.

[0460] 0.05% trifluoroacetic acid acetonitrile solution: 2 L of acetonitrile was measured out, 1 mL of trifluoroacetic acid was added, and the mixture was mixed uniformly and then ultrasonicated to obtain the solution.

[0461] SIF(E): 69.48 mg of potassium dihydrogen phosphate and 108.49 mg of pancreatin were weighed, 10 mL of purified water was added, and the mixture was mixed uniformly. The pH was then adjusted to 6.76 with 1 N aqueous sodium hydroxide solution (approximately 210 μL).

[0462] SGF(E): 21.64 mg of sodium chloride and 33.70 mg of pepsin were weighed, 10 mL of purified water was added, 20 μL of hydrochloric acid was added, and the pH was adjusted to 1.98 with 1 N aqueous sodium hydroxide solution (approximately 120 μL).

[0463] FaSSIF: 0.42 g of sodium hydroxide, 3.43 g of anhydrous sodium dihydrogen phosphate, and 6.19 g of sodium chloride were weighed, and 1 L of purified water was added. The mixture was sonicated and dissolved. The pH was then adjusted to 6.50 with 1 N sodium hydroxide or 1 N hydrochloric acid, and 2.24 g of 3F Powder was added and stirred until dissolved.

[0464] FeSSIF: 4.04 g of sodium hydroxide, 8.65 g of glacial acetic acid, and 11.87 g of sodium chloride were weighed, and 1 L of purified water was added. The mixture was sonicated and dissolved. The pH was then adjusted to 5.0 with 1 N sodium hydroxide or 1 N hydrochloric acid, and 11.2 g of 3F Powder was added and stirred until dissolved.

[0465] 7.3.2 Stability measurements Approximately 2 mg of compound was weighed, added to an appropriate amount of purified water, and dissolved by sonication to prepare a compound stock solution. 50 μL of the stock solution was taken, and 950 μL of SGF, FaSSIF, FeSSIF, SGF(E), and SIF(E) were added, respectively, and mixed uniformly. The sample stability was then examined at different time points.

[0466] 7.4 Liquid phase analysis: The liquid chromatography conditions were as follows:

[0467] [Table 63]

[0468] 7.5 Test Results and Analysis The compounds of the preferred examples of the present invention have excellent stability in simulated gastrointestinal fluids.

[0469] Test Example 8: Consideration of in vitro metabolic stability of the compounds of the present invention in liver microsomes 8.1 Study Objective: The metabolic stability of the compound of this example in mouse, rat, dog and human liver microsomes was investigated in phase I and partial phase II.

[0470] 8.2 Experimental Reagents 8.2.1 Reagents Compound of the present invention (self-prepared), liver microsomes (H0610 / M1000 / D1000, Xenotech), phosphate buffer (Lot#SLBS7904 and Lot#SLBR3106V, pH 7.4, Gibco), NADPH (reduced nicotinamide adenine dinucleotide phosphate, Shanghai Pinde Pharmaceutical Technology Co., Ltd.), UDPGA (Sigma), alamethicin (Lot#GR3226732-1, Paragon Pharmaceuticals), methanol (Merck), acetonitrile (Merck), DMSO (Sigma), 7-hydroxycoumarin (Paragon Pharmaceuticals).

[0471] 8.2.2 Drug Combinations Test compounds were formulated in DMSO to a 10 mM stock solution and stored in a refrigerator at -20°C until ready for use.

[0472] 8.3 Experimental steps 1) Buffer preparation 4.01 mL of 1 M K2HPO4·PO2O (AR grade) and 0.99 mL of 1 M KH2PO4 (AR grade) were taken and dissolved in ultrapure water, and the volume was adjusted to 50 mL to prepare a phosphate buffer solution with a final concentration of 100 mM.

[0473] 2) Preparation of compound working solution Preparation of compound working solution: 1 μL of compound stock solution was added to 999 μL of phosphate buffer to give a final concentration of 10 μM. Depending on the properties of the compound, the mixing ratio may be adjusted appropriately to adjust the final concentration.

[0474] 3) Preparation of liver microsomal working solution 156.3 μL of 20 mg / mL microsomes were diluted to 5 mL with 100 mM phosphate buffer and mixed uniformly to a final concentration of 0.625 mg / mL.

[0475] 4) Preparation of NADPH and UDPGA 33.3 mg of NADPH and 25.8 mg of UDPGA were weighed out and added to 2 mL of 100 mM phosphate buffer, with the final concentrations of both being 20 mM.

[0476] 5) Preparation of hole-punching agent (Alamethicin) 1 mg of Alamethicin was weighed out and added to 200 μL of methanol to prepare a 5 mg / mL solution, and 10 μL of this solution was then added to 990 μL of phosphate buffer (pH 7.4) to give a final concentration of 50 μg / mL.

[0477] 6) Preparation of reaction stop solution The internal standard was diluted with acetonitrile to give a final solution, which was then stored in a refrigerator at 2-8°C.

[0478] 7) Incubation process 400 μL of the prepared liver microsomes, 25 μL of compound working solution (10 μM), and 25 μL of Alamethicin (50 μg / mL) were added sequentially to a 96-well plate and pre-incubated at 37°C for 10 minutes. 50 μL of the prepared NADPH / UDPGA was then added to initiate the reaction, which was then incubated at 37°C. The total volume of the reaction system was 500 μL, and the final contents of each component were as follows: the compound of this example (0.5 μM), liver microsomes (0.5 mg / mL), NADPH (1 mM), UDPGA (1 mM), and Alamethicin (2.5 μg / mL).

[0479] At 0, 5, 15, 30, 60, and 120 min, 50 μL of the solution was removed, and 200 μL of cold stop solution containing an internal standard was added to stop the reaction. The sample was then centrifuged at 3500 rpm for 10 min, and the supernatant was collected and analyzed by LC-MS / MS.

[0480] 8.4 Biological analysis 1) Chromatography conditions Equipment: Shimadzu LC-20 AD Chromatography column: Phenomenex Gemiu™ C18 (50 x 4.6 mm, 5 μm particle size) Mobile phase: A: acetonitrile, B: 0.1% formic acid solution Flash gradient: 0.2-1.6 min 5%A-95%A, 3.0-3.1 min 95%A-5%A Flow rate: 1.0ml / min Run time: 4.0 min Sample input volume: 5 μL

[0481] 2) Mass spectrometry conditions Instrument: API4000 liquid chromatography mass spectrometer, AB Sciex Ion source: Electrospray ionization source (ESI) Drying gas: N2, temperature 500℃ Electrospray voltage: 5000V Detection method: Cation detection Scanning method: Multi-reaction monitoring (MRM) method Scan time: 0.8401s

[0482] 8.5 Data Processing The raw data was calculated using the following formula: Residual rate % = Peak area ratio of compound to internal standard at any time point / Peak area ratio of compound to internal standard at 0 minutes × 100% T 1 / 2 =0.693 / K e , where K e represents the removal rate constant.

[0483] K e In vitro hepatic microsome intrinsic clearance (CL) int ) and hepatic intrinsic clearance (CL int,liver ) calculation

[0484] CL int =0.693 / T 1 / 2 / microsomal protein content (microsomal concentration at incubation mg / mL)

[0485] CL int,liver =CL int nt Amount of microsomal protein in liver (mg / g) × × liver-to-body weight ratio

[0486] In vivo hepatic clearance (CL) was estimated based on a well-stirred model. CL=(CL int,liver nt,liver rate) / (CL int,liver nt, liver rate), where fu represents the free score in blood, default is 1.

[0487] The parameters in the formula are shown in the table below. [Table 64]

[0488] 8.6 Experimental Conclusions: The results showed that the compounds of the preferred examples of the present invention all exhibited stable metabolic effects in liver microsomes of various genera and species.

[0489] Test Example 9: Metabolic stability of the compound of the present invention in liver / kidney tissue homogenates 9.1 Experimental Objective: This experiment was conducted to determine the metabolic stability of the compound of this example in rat, dog, and monkey liver / kidney tissues.

[0490] 9.2 Experimental Reagents Rat liver tissue homogenate (Yakumei Kotoku), monkey liver tissue homogenate (Yakumei Kotoku), dog liver tissue homogenate (Yakumei Kotoku), human liver tissue homogenate (Bioreclamation IVT, S06585), rat kidney tissue homogenate (Yakumei Kotoku), monkey kidney tissue homogenate (Yakumei Kotoku), dog kidney tissue homogenate (Yakumei Kotoku), human kidney tissue homogenate (Bioreclamation IVT, S06585).

[0491] 9.3 Experimental steps 1) Solution preparation Test compound working solution: Dilute 5 μL of compound stock solution (10 mM dimethyl sulfoxide) with 995 μL of dimethyl sulfoxide (working solution concentration: 50 μM, 100% DMSO). Control Deslorelin working solution: Dilute 5 μL of Deslorelin stock solution (10 mM in dimethyl sulfoxide (DMSO)) with 495 μL of dimethyl sulfoxide (working solution concentration: 100 μM, 100% DMSO). Control semaglutide working solution: 50 μL of semagulide stock solution (1 mM dimethyl sulfoxide solution) was diluted with 450 μL of dimethyl sulfoxide (working solution concentration: 100 μM, 100% DMSO).

[0492] 2) Incubation process Prior to the experiment, the combined frozen liver / kidney homogenates were thawed in a 37°C water bath.

[0493] A 98 μL / well aliquot of blank liver / kidney homogenate was added to all 96-well reaction plates (blank, T0, T10, T30, T60, and T120). A 2 μL / well aliquot of working solution (50 or 100 μM) was added to all reaction plates except the blank (T0, T10, T30, T60, and T120). All reaction plates containing compounds and liver / kidney homogenate mixtures were incubated in a 37°C water bath.

[0494] The reaction plate was incubated at 37°C and a timer was started.

[0495] At the end of the incubation, 100 μL of sample was mixed with 100 μL of 4% H3PO4, and 800 μL of stop solution (methanol with tolbutamide and labetalol as internal standards) was added to precipitate proteins. Mix thoroughly.

[0496] Each plate was sealed and shaken for 20 minutes.

[0497] After shaking, each plate was centrifuged at 4000 rpm and 4°C for 20 minutes.

[0498] After centrifugation, 150 μL of supernatant was transferred from each reaction plate to its corresponding bioacylation plate.

[0499] Prior to LC-MS / MS analysis, each bioanalytical plate was sealed and shaken for 10 minutes.

[0500] 9.4 Data Processing The percentage remaining after the test compound was incubated in the liver / kidney homogenate was calculated by the following equation:

[0501] % remaining = 100 x (PAR incubation time point / PAR at TO time), where PAR is the peak area ratio between the analyte and the internal standard (is), and the incubation time points were TO (0 min), Tn (n = 0, 10, 30, 60, 120 min).

[0502] 9.5 Experimental Conclusions The results showed that the compounds of the preferred examples of the present invention have excellent stability in liver and kidney tissue homogenates.

[0503] Test Example 10: Consideration of the stability of the compound of the present invention in plasma 10.1 Research Objective: The stability of the compound of this example in mouse, rat, dog, and human plasma was investigated.

[0504] 10.2 Experimental Equipment Centrifuge (Eppendorf 5804R / 5424R), vortex machine (IKA VORTEX GENIUS 3), pipettor (Eppendorf 10-100 μL, Eppendorf 100-1000 μL, RAININ 0.5-10 μL), water bath (Shanghai Hengke).

[0505] 10.3 Experimental steps 1) Solution preparation Plasma production: After collecting whole blood from animals or humans, it is placed in a test tube containing an anticoagulant and centrifuged at 3500 rpm for 10 minutes, and the upper layer of pale yellow plasma is collected. 10 μM test compound working solution: Stock solution formulated in DMSO, working solution formulated in 100 mM phosphate buffer.

[0506] 10 μM positive control working solution: Procaine: Weigh out 2.36 mg of procaine and dilute it to a 10 mM stock solution with 1 mL of DMSO. Transfer 10 μL of the 10 mM stock solution to 1 mL of 100 mM phosphate buffer, so that the final concentration is 100 μM. Enalapril: 4.93 mg of enalapril was weighed and diluted with 1 mL of DMSO to a 10 mM stock solution, and 10 μL of the 10 mM stock solution was transferred to 1 mL of 100 mM phosphate buffer for a final concentration of 100 μM.

[0507] 2) Incubation process 285 μL of plasma and 15 μL of 10 μM compound (test compound, positive control) were added sequentially to a 96-well plate, and the plate was incubated at 37°C.

[0508] At 0, 15, 30, 60, 90, and 120 min (time points were adjustable), 40 μL was removed and 160 μL of acetonitrile stop solution containing an internal standard was added.

[0509] After centrifugation (3500 rpm, 10 min), 50 μL of the supernatant was collected and diluted with 50 μL of DDH2O before the sample was loaded onto the LC-MS / MS.

[0510] 10.4 Biological analysis 1) Chromatography conditions Equipment: Shimadzu LC-20 AD Chromatography column: Phenomenex Gemiu™ C 18 (50×4.6mm, 5μm particle size) Mobile phase: A: acetonitrile, B: 0.1% formic acid solution Flash gradient: 0-8 min: 5% A → 95% A, 2.0-2.1 min: 90% A → 5% A Flow rate: 0.8mL / min Run time: 5.0 min Sample input volume: 5 μL

[0511] 2) Mass spectrometry conditions Equipment: API4000 liquid chromatography mass spectrometer, AB, USA Ion source: Electrospray ionization source (ESI) Drying gas: N2, temperature 500℃ Electrospray voltage: 5500V Detection method: Cation detection Scanning method: Multi-reaction monitoring (MRM) method Scanning time: 0.1s

[0512] 10.5 Experimental results and data processing All calculations were performed in Microsoft Excel. Peak area ratios were determined from extracted ion chromatograms.

[0513] The percentage of compound remaining at each time point was calculated by the following equation: Remaining percentage t time (%) = Peak area ratio t time / Peak area 0 time × 100%

[0514] In the formula, the peak area ratio t hr is the peak area ratio between the control compound and the test compound at t min, The slope value k was determined by linear regression of the percent parent drug remaining versus the natural logarithm of the incubation time curve.

[0515] In vitro half-life (in vitro t 1 / 2 ) was determined by the slope. In vitro 1 / 2 =-(0.693 / k)

[0516] [Table 65]

[0517] 10.6 Experimental Conclusions: The compounds of the preferred examples of the present invention showed excellent stability in mouse, rat, dog and human plasma.

[0518] Test Example 11: In vivo pharmacodynamic study of the compounds of the present invention in an IL-23-induced rat otitis model 11.1 Experimental Objective: The in vivo efficacy of the compounds was evaluated in an IL23-induced rat otitis model.

[0519] 11.2 Laboratory equipment and reagents 11.2.1 Experimental equipment Refrigerator (BCD-268TN, Haier), biological safety cabinet (BSC-1300II A2, Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory), clean room (CJ-2F, Suzhou Feng's Laboratory Animal Equipment Co., Ltd.), electronic pipette (Easypet 3, Eppendorf), constant temperature water bath (HWS-12, Shanghai Yiheng Scientific), ultrasonic cleaner (115F0032, Shanghai Science and Technology), water purifier (Pacific TII, Thermo), magnetic stirrer (08-2G, Chikyu), electronic balance (CPA2202S, Sartorius), electronic balance (BSA2202S-CW, Sartorius), ultrasonic cell disrupter (JY92-IIN, Ningbo Xinzhi), micrometer (MDC-25PX, Sanfeng).

[0520] 11.2.2 Experimental Reagents IL-23 Protein, Rat, Recombinant (CT045-R08H, Sino Biological), PBS (10010-049, gibco), penetration enhancer, isoflurane (R510-22-10, Ruiwode), 4% tissue cell fixative (AR-0211, Dingguo).

[0521] 11.3 Experimental steps 11.3.1 Animal Purchase Male SD rats, 6-8 weeks old, were purchased from Jiangsu Jisui Yaokang Biotechnology Co., Ltd.

[0522] 11.3.2 Establishment of otitis model a, After the animals were acclimatized for one week, their tail numbers were marked with a felt-tip pen, and the animals were weighed and then randomly assigned to groups. b) On the day before model creation (i.e., D-1), animals were administered intragastric administration according to the experimental design and grouping. c,rrIL-23 was diluted with PBS to a concentration of 50 μg / ml, dispensed into 1 ml centrifuge tubes, and stored at −80°C until ready for use. d. On the first day of model creation (i.e., D0), the animals were administered intragastric administration according to the experimental design and grouping. Half an hour later, the rats were anesthetized with isoflurane, and after measuring the initial thickness of the right ear, IL-23 (1 μg, 20 μL) or 20 μL of PBS was intradermally injected into the right ear. e. On the second day (i.e., D1) of model creation, the animals were subjected to intragastric administration according to the experimental design and grouping. Half an hour later, the rats were anesthetized with isoflurane. After measuring the initial thickness of the right ear, IL-23 (1 μg, 20 μL) or 20 μL of PBS was intradermally injected into the right ear. f. On the third day (i.e., D2) of model creation, the animals were administered intragastric administration according to the experimental design and grouping. Half an hour later, the rats were anesthetized with isoflurane. After measuring the initial thickness of the right ear, IL-23 (1 μg, 20 μL) or 20 μL of PBS was intradermally injected into the right ear. g. On the fourth day (i.e., D3) of model creation, the animals were subjected to intragastric administration according to the experimental design and grouping. Half an hour later, the rats were anesthetized with isoflurane. After measuring the initial thickness of the right ear, IL-23 (1 μg, 20 μL) or 20 μL of PBS was intradermally injected into the right ear. On the fifth day (i.e., D4) of the experiment, the final thickness of the right ear was measured. According to the experimental design and grouping, the animals were intragastricly administered. One hour later, the animals were euthanized and blood and ear tissue samples were collected. The ears were divided into two parts: one part was snap-frozen and used for inflammatory factor mRNA detection, and the other part was fixed in 4% tissue cell fixative for histological processing and analysis (HE staining). The plasma was used for PK detection.

[0523] 11.4 Data Processing: a) The initial ear thickness was subtracted from the daily ear thickness of animals in each group (ear thickness increase) and compared with the model group (vehicle).

[0524] b) Ear thickness results from Day 4 to Day 0, the final day of the experiment, were plotted and analyzed using Graphpad Prism 9. One-way ANOVA Dunnett's test was used to analyze differences between groups in ear thickness changes, and p<0.05 was considered significant. Inhibition rate of ear thickness (%) = [1 - (mean value of treatment group / mean value of model group)] × 100%.

[0525] 11.5 Experimental Results: [Table 66]

[0526] 11.6 Experimental Conclusion: In an IL23-induced rat otitis model, the compounds of the preferred embodiments of the present invention were able to effectively inhibit the increase in ear thickness, and once-daily administration had a more significant inhibitory effect on ear thickness increase than twice-daily administration of PN-235, achieving the purpose of extending the administration cycle.

Claims

1. A compound represented by general formula (I), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 【Chemistry 1】 R １ and R ２ are each independently H, an alkyl group, 【Chemistry 2】 or an amino acid, R ａ is H, an alkyl group, 【Transformation 3】 or an amino acid, R ａａ is an alkyl group, 【Chemistry 4】 or an amino acid, X １ is selected from amino acids, Preferably, R ａａ is an alkyl group, 【Transformation 5】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ and R ７ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Transformation 6】 or an amino acid, R ｃ is H, an alkyl group, 【Transformation 7】 or an amino acid, R ｃｃ is an alkyl group, 【Transformation 8】 or an amino acid, X ５ is selected from amino acids, X ６ is selected from amino acids, X ７ is selected from amino acids, Preferably, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 9】 or an amino acid, R ７ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 10】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 11】 or an amino acid, R ｃｃ is an alkyl group, 【Chemistry 12】 or an amino acid, X ５ is selected from amino acids, X' ５ is selected from amino acids, X ６ is selected from amino acids, X' ６ is selected from amino acids, X ７ is selected from amino acids, X' ７ is selected from amino acids, R ４ and R ５ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Chemistry 13】 or an amino acid, R ｂ is H, an alkyl group, 【Chemistry 14】 or an amino acid, R ｂｂ is an alkyl group, 【Chemistry 15】 or an amino acid, X ２ is selected from amino acids, X ３ is selected from amino acids, X ４ is selected from amino acids, Preferably, R ４ represents H, an alkyl group, or —CH ２ C(O)NH ２ , 【Chemistry 16】 or an amino acid, R ５ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 17】 or amino acids, preferably [Chemistry 18] is selected from R ｂ is H, an alkyl group, 【Chemistry 19】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is an alkyl group, 【Chemistry 20】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n to n2 are each independently selected from integers of 0 to 12; t to t9 are each independently selected from integers of 0 to 12; m to m21 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; The compound is 【Chemistry 21】 Instead, Preferably, R １ and R ２ are each independently H, an alkyl group, 【Chemistry 22】 or an amino acid, R ａ is H, an alkyl group, 【Chemistry 23】 or an amino acid, R ａａ is an alkyl group, 【Chemistry 24】 or an amino acid, X １ is selected from amino acids, Preferably, R ａａ is an alkyl group, 【Chemistry 25】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, R ３ is selected from a hydroxy group, R ４ and R ５ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ is selected from n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24; Or, R １ and R ２ is selected from H, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ and R ７ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Chemistry 26】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 27】 or an amino acid, R ｃｃ is an alkyl group, 【Chemistry 28】 or an amino acid, X ５ is selected from amino acids, X ６ is selected from amino acids, X ７ is selected from amino acids, Preferably, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 29】 or an amino acid, R ７ represents H, an alkyl group, or —CH ２ C(O)NH ２ , 【Transformation 30】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 31】 or an amino acid, R ｃｃ is an alkyl group, 【Chemistry 32】 or an amino acid, X ５ is selected from amino acids, X' ５ is selected from amino acids, X ６ is selected from amino acids, X' ６ is selected from amino acids, X ７ is selected from amino acids, X' ７ is selected from amino acids, R ４ and R ５ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ is selected from n2 is selected from integers from 0 to 12; t6 to t9 are each independently selected from integers of 0 to 12; m13 to m21 are each independently selected from integers of 0 to 24; Or, R １ and R ２ is selected from H, R ３ is selected from a hydroxy group, R ４ and R ５ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Transformation 33】 or an amino acid, R ｂ is H, an alkyl group, 【Transformation 34】 or an amino acid, R ｂｂ is an alkyl group, 【Chemistry 35】 or an amino acid, X ２ is selected from amino acids, X ３ is selected from amino acids, X ４ is selected from amino acids, Preferably, R ４ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Transformation 36】 or an amino acid, R ５ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 37】 or amino acids, preferably 【Transformation 38】 is selected from R ｂ is H, an alkyl group, 【Chemistry 39】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is an alkyl group, 【Chemistry 40】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers of 0 to 24, a compound represented by formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

2. A compound represented by general formula (IV), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 【Chemistry 41】 X １ is selected from amino acids, X ２ is selected from amino acids, R １ and R ２ are each independently H, an alkyl group, 【Chemistry 42】 or an amino acid, R ａ is H, an alkyl group, 【Chemistry 43】 or an amino acid, R ａａ is an alkyl group, 【Chemistry 44】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 45】 or an amino acid, R ７ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 46】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 47】 or an amino acid, R ｃｃ is an alkyl group, 【Chemistry 48】 or an amino acid, X ５ is selected from amino acids, X' ５ is selected from amino acids, X ６ is selected from amino acids, X' ６ is selected from amino acids, X ７ is selected from amino acids, X' ７ is selected from amino acids, R ４ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 49】 or an amino acid, R ５ is H, an alkyl group, -CH ２ C(O)NH ２ , [Transformation 50] or an amino acid, R ｂ is H, an alkyl group, 【Chemistry 51】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is an alkyl group, 【Chemistry 52】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n to n2 are each independently selected from integers of 0 to 12; t to t9 are each independently selected from integers of 0 to 12; m to m21 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; The compound is 【Chemistry 53】 A compound represented by general formula (IV), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is not

3. The compound may further be as shown in general formula (II) or (III): 【Chemistry 54】 where: R １ and R ２ are each independently H, an alkyl group, 【Transformation 55】 or an amino acid, R ａ is H, an alkyl group, 【Transformation 56】 or an amino acid, R ａａ is an alkyl group, 【Chemistry 57】 or an amino acid, X １ is selected from amino acids, Preferably, R ａａ is an alkyl group, 【Chemistry 58】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ and R ７ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Chemistry 59】 or an amino acid, R ｃ is H, an alkyl group, 【Transformation 60】 or an amino acid, R ｃｃ is an alkyl group, 【Chemistry 61】 or an amino acid, X ５ is selected from amino acids, X ６ is selected from amino acids, X ７ is selected from amino acids, Preferably, R ３ represents a hydroxy group, an alkyl group, or —NR ６ R ７ or an amino acid, R ６ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Transformation 62】 or an amino acid, R ７ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Transformation 63】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 64】 or an amino acid, R ｃｃ is an alkyl group, 【Transformation 65】 or an amino acid, X ５ is selected from amino acids, X' ５ is selected from amino acids, X ６ is selected from amino acids, X' ６ is selected from amino acids, X ７ is selected from amino acids, X' ７ is selected from amino acids, R ４ is selected from H or an alkyl group; n is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; m is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; n2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t6 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t7 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t8 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t9 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; m13 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m14 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m15 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m16 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m17 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m18 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m19 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m20 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; The compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, characterized in that m21 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24.

4. R １ is H, C １－６ alkyl groups, 【Chemical Formula 66】 or an amino acid, R ａ is H, C １－６ alkyl groups, 【Transformation 67】 or an amino acid, R ａａ is C １－６ alkyl groups, 【Transformation 68】 or an amino acid, X １ is selected from amino acids, Preferably, R ａａ is C １－６ alkyl groups, 【Transformation 69】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24; and / or R ２ is H, C １－６ alkyl groups, 【Transformation 70】 or an amino acid, R ａ is H, C １－６ alkyl groups, 【Chemistry 71】 or an amino acid, R ａａ is C １－６ alkyl groups, 【Chemistry 72】 or an amino acid, X １ is selected from amino acids, Preferably, R ａａ is C １－６ alkyl groups, 【Transformation 73】 or an amino acid, X １ is selected from amino acids, X' １ is selected from amino acids, n is selected from an integer from 0 to 12; t to t1 are each independently selected from integers of 0 to 12; m to m3 are each independently selected from integers of 0 to 24; and / or R ３ is a hydroxy group, C １－６ alkyl group, -NR ６ R ７ or an amino acid, R ６ and R ７ are each independently H, an alkyl group, or —CH ２ C(O)NH ２ , 【Chemistry 74】 or an amino acid, R ｃ is H, an alkyl group, 【Chemistry 75】 or an amino acid, R ｃｃ is an alkyl group, 【Transformation 76】 or an amino acid, X ５ is selected from amino acids, X ６ is selected from amino acids, X ７ is selected from amino acids, Preferably, R ３ is a hydroxy group, C １－６ alkyl group, -NR ６ R ７ or an amino acid, R ６ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Chemical Formula 77】 or an amino acid, R ７ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Transformation 78】 or an amino acid, R ｃ is H, C １－６ alkyl groups, 【Transformation 79】 or an amino acid, R ｃｃ is C １－６ alkyl groups, 【Chemistry 80】 or an amino acid, X ５ is selected from amino acids, X' ５ is selected from amino acids, X ６ is selected from amino acids, X' ６ is selected from amino acids, X ７ is selected from amino acids, X' ７ is selected from amino acids, n2 is selected from integers from 0 to 12; t6 to t9 are each independently selected from integers of 0 to 12; The compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 2, wherein m13 to m21 are each independently selected from integers of 0 to 24.

5. The compound is further represented by general formula (V): 【Chemistry 81】 where: X １ is selected from amino acids, X ２ is selected from amino acids, R ４ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 82】 or an amino acid, R ５ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 83】 or an amino acid, R ｂ is H, an alkyl group, 【Chemical 84】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is an alkyl group, 【Chemical 85】 or an amino acid, X' ２ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 12; The compound is 【Chemical 86】 3. The compound of claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof, which is not:

6. X １ , Thr, 【Chemistry 87】 is selected from, preferably 【Chemical 88】 and and / or X ２ teeth, 【Chemical 89】 is selected from, preferably 【Chemistry 90】 3. The compound according to claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein:

7. The compound is further represented by general formula (IV): 【Chemistry 91】 where: R ４ is selected from H or an alkyl group; R ５ is an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 92】 or an amino acid, R ｂ is H, an alkyl group, 【Chemistry 93】 or an amino acid, R ｂｂ is an alkyl group, 【Chemical 94】 or an amino acid, X ２ is selected from amino acids, X ３ is selected from amino acids, X ４ is selected from amino acids, Preferably, R ４ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemical 95】 or an amino acid, R ５ is H, an alkyl group, -CH ２ C(O)NH ２ , 【Chemistry 96】 or amino acids, preferably 【Chemistry 97】 is selected from R ｂ is H, an alkyl group, 【Chem.98】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is an alkyl group, 【Chem.99】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n1 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t2 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t3 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t4 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; t5 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12; m4 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m5 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m6 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m7 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m8 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m9 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m10 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m11 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; m12 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, or 24; The compound, its stereoisomer or pharmaceutically acceptable salt thereof according to claim 1, characterized in that m22 is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24.

8. R ４ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Chemistry 100】 or an amino acid, R ｂ is H, C １－６ alkyl groups, 【Chemistry 101】 or an amino acid, R ｂｂ is C １－６ alkyl groups, 【Chemical Engineering 102】 or an amino acid, X ２ is selected from amino acids, X ３ is selected from amino acids, X ４ is selected from amino acids, Preferably, R ４ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Chemistry 103】 or an amino acid, R ｂ is H, C １－６ alkyl groups, 【Chemical 104】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is C １－６ alkyl groups, 【Chemistry 105】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; and / or R ５ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Chemistry 106】 or an amino acid, R ｂ is H, C １－６ alkyl groups, 【Chemistry 107】 or an amino acid, R ｂｂ is C １－６ alkyl groups, 【Chemistry 108】 or an amino acid, X ２ is selected from amino acids, X ３ is selected from amino acids, X ４ is selected from amino acids, Preferably, R ５ is H, C １－６ Alkyl group, —CH ２ C(O)NH ２ , 【Chemistry 109】 or amino acids, more preferably 【Chemical 110】 is selected from R ｂ is H, C １－６ alkyl groups, 【Chemistry 111】 or an amino acid, wherein the alkyl group may be further substituted with a hydroxy group or an alkoxy group; R ｂｂ is C １－６ alkyl groups, 【Chemistry 112】 or an amino acid, X ２ is selected from amino acids, X' ２ is selected from amino acids, X ３ is selected from amino acids, X' ３ is selected from amino acids, X ４ is selected from amino acids, X' ４ is selected from amino acids, n1 is selected from integers from 0 to 12; t2 to t5 are each independently selected from integers of 0 to 12; m4 to m12 are each independently selected from integers of 0 to 24; m22 is selected from integers from 0 to 24; Preferably, 【Chemistry 113】 teeth, 【Chemical 114】 is selected from, preferably 【Chemical 115】 is selected from 【Chemistry 116】 teeth, 【Chemistry 117】 is selected from, preferably 【Chemistry 118】 3. The compound according to claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof, selected from:

9. The following compound, its stereoisomer, or a pharmaceutically acceptable salt thereof: 【Chemical 119】 【Chemical 120】 【Chemistry 121】 【Chemistry 122】 【Chemical 123】 【Chemistry 124】 【Chemistry 125】 【Chemistry 126】 A compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that:

10. A process for preparing the compound of claim 2, its stereoisomer or its pharmaceutically acceptable salt, which is based on solid phase or liquid phase synthesis, Preferably, the synthesis method comprises: Step 1) synthesizing a resin peptide based on a solid phase synthesis method; Step 2) of cleaving the resin peptide obtained in step 1) to obtain a polypeptide intermediate; Step 3) condensing the polypeptide intermediate with the side chain; and step 4) removing the protecting groups from the peptide segment obtained in step 3) and then forming a ring to obtain a final product, More preferably, the synthesis method comprises: Step 1) synthesizing a resin peptide based on the solid phase synthesis method of the Fmoc method and end-capping it with acetic anhydride; Step 2) of cleaving the resin peptide obtained in step 1) to obtain a polypeptide intermediate; Step 3) condensing the polypeptide intermediate with the side chain using a coupling agent; and step 4) removing the protecting groups from the peptide segment obtained in step 3) and then forming a ring by oxidation of the disulfide bond to obtain a final product.

11. A pharmaceutical composition comprising a therapeutically effective amount of a compound of claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

12. 10. The compound of claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof for use in inhibiting IL-23R in a subject in need thereof.

13. Inflammatory and autoimmune diseases and cancers, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome and Wiskott-Aldrich syndrome, following proctocolectomy and ileal pouch-anal anastomosis.

10. The compound according to claim 2, its stereoisomer or a pharmaceutically acceptable salt thereof for use in the treatment of pouchitis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), plaque psoriasis, plaque psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetic diseases, preferably inflammatory bowel disease (IBD), rheumatoid arthritis or plaque psoriasis.

14. The pharmaceutical composition of claim 11 for use in inhibiting IL-23R in a subject in need thereof.

15. Inflammatory and autoimmune diseases and cancers, such as inflammatory bowel disease (IBD), ulcerative colitis, Crohn's disease, celiac disease (non-tropical sprue), enteropathy associated with seronegative arthritis, microscopic colitis, collagenous colitis, eosinophilic gastroenteritis, colitis associated with radiation therapy or chemotherapy, colitis associated with congenital immune diseases such as leukocyte adhesion deficiency-1, chronic granulomatous disease, glycogen storage disease type 1b, Hermansky-Pudlak syndrome, Chediak-Higashi syndrome and Wiskott-Aldrich syndrome, proctocolectomy and ileal pouch anal 12. The pharmaceutical composition according to claim 11 for use in the treatment of pouchitis following porto-hilar anastomosis, gastrointestinal cancer, pancreatitis, insulin-dependent diabetes mellitus, mastitis, cholecystitis, cholangitis, pericholangitis, chronic bronchitis, chronic sinusitis, asthma, psoriasis, psoriatic arthritis, irritable bowel syndrome (IBS), multiple sclerosis (MS), plaque psoriasis, plaque psoriatic arthritis, rheumatoid arthritis, pemphigus vulgaris, organ transplant rejection, Crohn's disease, systemic lupus erythematosus (SLE) or diabetic diseases, preferably inflammatory bowel disease (IBD), rheumatoid arthritis or plaque psoriasis.

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