Interleukin receptor antagonist and application thereof

Abstract

Provided are an interleukin receptor antagonist and an application thereof, and specifically provided is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The provided compound exhibits potent biological activity and good pharmacokinetic properties, or demonstrates good efficacy in animal pharmacodynamic studies. c[X4-X5-X6-X7-X8-X9]-X10-X11-X12-X13-X14-X15-X16 (I)

Description

An interleukin receptor antagonist and its application

[0001] This application claims priority to Chinese patent applications filed on December 18, 2024 (202411872806X), December 26, 2024 (2024119399731), March 21, 2025 (2025103430531), October 30, 2025 (2025115724216), November 21, 2025 (2025117230028), and December 11, 2025 (2025118758835). The full text of the aforementioned Chinese patent applications is incorporated herein by reference. Technical Field

[0002] This invention relates to an interleukin receptor antagonist and its application. Background Technology

[0003] Interleukin-23 (IL-23) is a heterodimeric hematopoietic cytokine composed of two subunits, p40 and p19. Structurally, it shares the p40 subunit with IL-12, and they are linked by disulfide bonds. The IL-23 receptor is composed of two subunits: the IL-12 receptor β1 subunit (IL-12Rβ1) and the IL-23 receptor (IL-23R). IL-23R is a unique subunit of the IL-23 pathway and interacts with the IL-23p19 subunit. The IL-23R protein contains an extracellular region, a single transmembrane region, and a cytoplasmic region of 252 amino acids. Unlike the three membrane fibrin junctions of other hematopoietic factor receptor superfamily members, its extracellular region consists of a signal sequence, an N-terminal immunoglobulin-like domain, and two cytokine receptor regions.

[0004] IL-23 is a cytokine with multiple biological functions, acting on various cell types to promote cytokine secretion and produce corresponding cellular effects. IL-23 can act on human memory T cells, promoting their vigorous proliferation. IL-23 can induce T cells to produce various cytokines such as IFN-γ, IL-17, and IL-10. In addition to acting directly on T cells, IL-23 can also activate and regulate T cell-dependent immune responses through dendritic cells (DCs). It can regulate granulocyte production by modulating neutrophil regulatory cells to control IL-17 secretion.

[0005] IL-23 provides essential conditions for the generation and survival of Th17 cells. Th17 is a recently discovered subset of CD4+ T cells, distinct from Th1 and Th2 cells. This subset highly expresses IL-17 and also produces IL-6 and TNF-α. IL-17 is an inflammatory cytokine whose primary biological function is to promote inflammatory responses, and it is closely related to autoimmune diseases such as rheumatoid arthritis, asthma, multiple sclerosis, and psoriasis, as well as transplant rejection. Th17 cells mediate the occurrence and development of inflammatory responses, autoimmune diseases, tumors, and transplant rejection. The differentiation and function of the Th17 cell subset are regulated by Th1 and Th2 cytokines. Studies have confirmed that IL-23 stimulates the expansion and maintenance of Th17 cells; therefore, IL-23R is a key factor for the Th17 cell subset. The pathogenicity of the IL23-IL23R-Th17-IL17 axis in autoimmune diseases has also been confirmed.

[0006] The global market for autoimmune disease drugs exceeds $100 billion. According to Frost & Sullivan, the global market is projected to reach $176.7 billion by 2030. In comparison, the global market for anti-tumor drugs was $239.8 billion in 2023. From a market size perspective, autoimmune diseases have the potential to become the next largest market after oncology drugs. Autoimmune diseases are difficult to cure completely, and traditional therapies have limited efficacy. The causes of most autoimmune diseases are not fully understood, and the alternation between flare-ups and remissions makes it difficult to cure them with current treatments. Traditional treatments have limited efficacy for patients with severe conditions and have significant side effects with long-term use. In contrast, targeted drugs can selectively suppress the immune system, thereby reducing side effects and acting faster. Currently, most targeted drugs in the autoimmune field are biological agents, primarily antibodies, administered via injection. For chronic diseases like autoimmune diseases, oral formulations are more convenient, leading to better patient compliance and drug accessibility. However, currently available oral small molecule drugs are less effective than biologics for autoimmune diseases and have more side effects, which greatly limits patients' choices of oral medications. There is an urgent clinical need for oral autoimmune drugs with better efficacy and fewer side effects. Summary of the Invention

[0007] To address the problems of poor pharmacokinetic properties in existing drugs for autoimmune diseases, this invention provides an interleukin receptor antagonist and its applications. This invention provides compounds that specifically block the IL-23 signaling pathway by blocking IL-23R, compositions comprising these molecules, methods for screening these molecules, and methods for using these molecules to treat various autoimmune diseases. These molecules possess one or more of the following advantages: (1) extremely strong biological activity; (2) good pharmacokinetic properties; (3) good efficacy in animal studies.

[0008] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0009] c[X4-X5-X6-X7-X8-X9]-X10-X11-X12-X13-X14-X15-X16

[0010] (I)

[0011] In c[X4-X5-X6-X7-X8-X9], “c[]” indicates that X4 and X9 are connected in a ring;

[0012] X6, X7, X8, X11, X12, X13, and X14 are any amino acid residues;

[0013] X4 is The carbonyl end is connected to X5;

[0014] R 4 -C(=O)R 4a , -L 1 -Z 1 or -L 2 -(L 3 ) n1 -(L 4 ) n1a -Z 2 ;

[0015] R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0016] X5 is The carbonyl end is connected to X6;

[0017] R 5a C 1-6 Alkyl, -L 1 -Z 1 Or be -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 alkyl;

[0018] R 5a-1 and R 5a-2 Independently H or C 1-6Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups;

[0019] R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0020] X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8; R 9a and R 9b Independently H or C 1-6 alkyl;

[0021] X10 is The carbonyl end is connected to X11;

[0022] R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy;

[0023] R 10a-1 and R 10a-2 Independently -NH2, -OH, or halogen;

[0024] Cycle B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or a ring with one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups;

[0025] R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen;

[0026] X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0027] X16 is

[0028] R 16 Independently -NH2, -L 1 -Z 1 -L 2 -(L 3 ) n1 -L 4 -Z 2 or -L 2 -(L 3 ) n1 -(L 4 ) n1a -Z 2 ;

[0029] And at least one of X4 and X16 contains -L 2 -(L 3 ) n1 -L 4 -Z 2 or -L 2 -(L 3 ) n1 -(L 4 ) n1a -Z 2 structure;

[0030] L 1 and L 2 Independent of chemical bonds or connecting units;

[0031] Z 1 Independently

[0032] Z 1a and Z 1c Independently -NH-, -C(=O)-, -O-, or -S-;

[0033] Z 1b H, -SF5, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0034] Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0035] L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -N(R L3 - or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -N(R L3 )-C(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 Alkylene-N(R) L3 )-;

[0036] R L3 For H or C 1-6 alkyl;

[0037] L 4 Independently

[0038] R L4a and R L4b H and C independently 1-6 alkyl or

[0039] Z 1g H, halogen or C 1-6 alkyl;

[0040] Z 2 Independently -C (=O) -C 6-24 Alkylene-R z ;R z -CO2H, -SO3H,

[0041] n1, n1a, n3, n4, n5, n6, n7, n8, n9, n9a, n9b, n10, n10a, n11, n12 and n13 are independently 0, 1, 2, 3, 4, 5 or 6;

[0042] In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0043] The condition is that the compound represented by formula (I) satisfies one, two, or three of the following conditions:

[0044] (1) X10 is

[0045] (2)X15 is

[0046] (3) X8 is

[0047] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0048] c[X4-X5-X6-X7-X8-X9]-X10-X11-X12-X13-X14-X15-X16

[0049] (I)

[0050] In c[X4-X5-X6-X7-X8-X9], “c[]” indicates that X4 and X9 are connected in a ring;

[0051] X6, X7, X8, X11, X12, X13, and X14 are any amino acid residues;

[0052] X4 is The carbonyl end is connected to X5;

[0053] R 4 -C(=O)R 4a , -L 1 -Z 1 or -L 2 -(L 3 ) n1 -L 4 -Z 2 ;

[0054] R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0055] X5 is The carbonyl end is connected to X6;

[0056] R 5a C1-6 Alkyl, -L 1 -Z 1 Or be -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 alkyl;

[0057] R 5a-1 and R 5a-2 Independently H or C 1-6 Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups;

[0058] R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0059] X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8; R 9a and R 9b Independently H or C 1-6 alkyl;

[0060] X10 is The carbonyl end is connected to X11;

[0061] R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy;

[0062] R 10a-1 and R 10a-2 Independently -NH2, -OH, or halogen;

[0063] Ring B is a 5-8 membered heterocyclic alkenyl group, or is surrounded by one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups;

[0064] R 10-1 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen;

[0065] X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0066] X16 is

[0067] R 16 Independently -NH2, -L 1 -Z 1 or -L 2 -(L 3 ) n1 -L 4 -Z 2 ;

[0068] And at least one of X4 and X16 contains -L 2 -(L 3 ) n1 -L 4 -Z 2 structure;

[0069] L 1 and L 2 Independent of chemical bonds or connecting units;

[0070] Z 1 Independently

[0071] Z 1a and Z 1c Independently -NH-, -C(=O)-, -O-, or -S-;

[0072] Z 1b H, -SF5, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0073] Z 1d Z 1eand Z 1f The site is H or C 1-6 alkyl;

[0074] L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -N(R L3 )-;

[0075] R L3 For H or C 1-6 alkyl;

[0076] L 4 Independently

[0077] R L 4 a and R L4b H and C independently 1-6 alkyl or

[0078] Z 1g H, halogen or C 1-6 alkyl;

[0079] Z 2 Independently -C (=O) -C 6-24 Alkylene -CO2H;

[0080] n1, n3, n4, n5, n6, n7, n8, n9, n10, n11, n12, and n13 are independently 0, 1, 2, 3, 4, 5, or 6;

[0081] In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0082] The condition is that the compound represented by formula (I) satisfies one, two, or three of the following conditions:

[0083] (1) X10 is

[0084] (2)X15 is

[0085] (3) X8 is

[0086] This invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0087] c[X4-X5-X6-X7-X8-X9]-X10-X11-X12-X13-X14-X15-X16

[0088] (I)

[0089] In c[X4-X5-X6-X7-X8-X9], “c[]” indicates that X4 and X9 are connected in a ring;

[0090] X6, X7, X8, X11, X12, X13, and X14 are any amino acid residues;

[0091] X4 is The carbonyl end is connected to X5;

[0092] R 4 -C(=O)C 1-6 alkyl, -L 1 -Z 1 or -L 2 -(L 3 ) n1 -L 4 -Z 2 ;

[0093] X5 is The carbonyl end is connected to X6;

[0094] R 5a C 1-6 Alkyl, -L 1 -Z 1 C that is replaced by -C(=O)NH2 1-6 alkyl;

[0095] X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8; R 9a and R9 b Independently H or C 1-6 alkyl;

[0096] X10 is The carbonyl end is connected to X11;

[0097] R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy;

[0098] R 10a-1 and R 10a-2Independently -NH2, -OH, or halogen;

[0099] R 10b For H or C 1-6 alkyl;

[0100] X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0101] X16 is

[0102] R 16 -NH2, -L 1 -Z 1 or -L 2 -(L 3 ) n1 -L 4 -Z 2 ;

[0103] And at least one of X4 and X16 contains -L 2 -(L 3 ) n1 -L 4 -Z 2 structure;

[0104] L 1 and L 2 Independent of chemical bonds or connecting units;

[0105] Z 1 Independently

[0106] Z 1a and Z 1c Independently -NH-, -C(=O)-, -O-, or -S-;

[0107] Z 1b H, -SF5, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C2-6 alkynyl group, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0108] Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0109] L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -N(R L3 )-;

[0110] R L3 For H or C 1-6 alkyl;

[0111] L 4 Independently

[0112] R L4a and R L4b H and C independently 1-6 alkyl or

[0113] Z 1g H, halogen or C 1-6 alkyl;

[0114] Z 2 Independently -C (=O) -C 6-24 Alkylene -CO2H;

[0115] n1, n2, n3, n4, n5, n6, n7, n8, n9, n10, n11, n12, and n13 are independently 0, 1, 2, 3, 4, 5, or 6;

[0116] In the 5-6 membered heteroaryl group, the heteroatom is one, two, or three of N, O, and S, and the number of heteroatoms is one, two, or three.

[0117] The condition is that the compound represented by formula (I) satisfies one or both of the following conditions:

[0118] (1) X10 is

[0119] (2)X15 is

[0120] In this invention, the amino acid residues include natural amino acid residues and non-natural amino acid residues, and contain carbonyl groups and nitrogen-containing groups.

[0121] In some embodiments, when the compound of formula (I) contains a cationic fragment, the anion is acetate, trifluoroacetate, chloride, bromide, adipic acid, benzoate, benzenesulfonate, citrate, decanoate, lactate, maleate, methanesulfonate, propionate, oxalate, succinate, sulfate, or tartrate; the cation and anion have equal charges.

[0122] In some implementations, L 1 and L 2 In this context, the connecting unit is a combination of one or more groups selected from the following: C 1-6 Alkylene, -C(=O)-, -NR La -、 -C(=O)NR Ld -、-NR Le C(=O)-, -C(=O)O-, -O-, -S-, -(CH2CH2O) m -、 Or by one or more R Lf Replacement C 1-6 Alkylene;

[0123] R La R Lb R Lc R Ld and R Le Independently H or C 1-6 alkyl;

[0124] R Lf C 1-6 Alkyl, -NHC(=NH)NH2 or -CONH2.

[0125] m can be 1, 2, 3, 4, 5, 6, 7 or 8.

[0126] In some implementations, L 1 Independent of chemical bonds, -C(=O)-C 1-6 alkylene-C(=O)- or a-end and Z 1 Connection; R Lb and R Lc Independently H or C 1-6 Alkyl group; m1, m2, m3 and m4 are independently 0, 1, 2, 3, 4 or 5.

[0127] In some implementations, L 1 Independently -C(=O)-(CH2CH2O) m2a -C 1-6 Alkylene NH- a a end and Z end 1Connection; m2a is 1-10.

[0128] In some implementations, L 1 Independent of chemical bonds, a-end and Z 1 connect.

[0129] In some implementations, L 1 Independently a-end and Z 1 connect.

[0130] In some implementation schemes, Z 1 for

[0131] In some implementation schemes, Z 1 for

[0132] In some implementations, L 2 Independently for -L 2a -C 1-6 Alkylene-L 2 b-、 -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1- 6-alkylene- or The C 1- 6-alkylene groups are optionally surrounded by one or more R Lf replace;

[0133] L 2a and L 2b Independently -C(=O)- or -NH-;

[0134] L 2c L 2d L 2e and L 2f Independently -C(=O)-, -CH2-, or -NH-;

[0135] m is 1, 2, 3, 4 or 5;

[0136] m5, m6, m7, m8, m9, m10, m11 and m12 are independently 0, 1, 2, 3, 4 or 5;

[0137] R Lb R Lc and R Lf The definition is as described above.

[0138] In some implementations, L 2 Independently L 2g and L 2h Independently -C(=O)-, -CH2-, or -NH-;

[0139] m13 is independently 0, 1, 2, 3, 4, or 5; m14, m15, and m16 are independently 1, 2, 3, 4, or 5; R Lb and R Lc The definition is as described above.

[0140] In some implementations, L 2 Independently for -L 2a -C 1-6 Alkylene-L 2b -、 -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1- 6-alkylene- The C 1-6 Alkylene is optionally surrounded by one or more R Lf replace;

[0141] L 2a and L 2b Independently -C(=O)- or -NH-;

[0142] L 2c L 2d L 2e L 2f L 2g and L 2h Independently -C(=O)-, -CH2-, or -NH-;

[0143] m is 1, 2, 3, 4 or 5;

[0144] m5, m6, m7, m8, m9, m10, m11, m12, and m13 are independently 0, 1, 2, 3, 4, or 5;

[0145] m14, m15 and m16 are independently 1, 2, 3, 4 or 5;

[0146] R Lb R Lc and R Lf The definition is as described above.

[0147] In some implementations, L 2 Independently -C (=O) -C 1-6 alkylene-NH- b -C(=O)-C 1-6 Alkylene-C(=O)- b -NH-C 1-6 alkylene-NH- b , -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 alkylene- b or The C 1- 6-alkylene compounds are optionally R Lf Replacement; b-end and L 3 connect;

[0148] m is 1, 2, 3, 4 or 5;

[0149] m5, m6, m7, m8, m9, m10, m11 and m12 are independently 0, 1, 2, 3, 4 or 5;

[0150] R Lb R Lc and R Lf The definition is as described above.

[0151] In some implementations, L 2 Independently b end and L 3 connect;

[0152] m13 is independently 0, 1, 2, 3, 4, or 5; m14, m15, and m16 are independently 1, 2, 3, 4, or 5; R Lb and R Lc The definition is as described above.

[0153] In some implementations, L 2 Independently -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 3 Connection; R Lf It is -CONH2.

[0154] In some implementations, L 2 Independent of chemical bonds, b end and L 3 connect.

[0155] In some implementations, L 2 Independently b end and L 3 connect.

[0156] In some implementations, L 2 Independently b end and L 3 connect.

[0157] In some implementations, L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH-; n1 is 0, 1 or 2.

[0158] In some implementations, L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 Alkylene-NH-.

[0159] In some implementations, n1 is 0, 1, or 2.

[0160] In some implementations, L 3 Independently c-end and L 4 Connect; n1 is 0, 1, or 2.

[0161] In some implementations, L 3 Independently c-end and L 4 connect.

[0162] In some implementations, L 4 Independently

[0163] R L4a and R L4b H and C independently 1-6 alkyl or

[0164] Z 1g H, halogen or C1-6 alkyl;

[0165] n10, n11, n12 and n13 are independently 1, 2, 3, 4 or 5.

[0166] In some implementations, L 4 Independently n10a can be 1, 2, 3, 4 or 5 independently.

[0167] In some implementations, L 4 for d end and Z end 2 connect.

[0168] In some implementations, L 4 for d end and Z end 2 connect.

[0169] In some implementation schemes, Z 2 for

[0170] In some implementation schemes, Z 2 for

[0171] In some implementations, X6, X7, X8, X11, X12, X13, and X14 are independently...

[0172] R a For H or C 1-6 alkyl;

[0173] R b -(CH2) p1 -R b-1 ;

[0174] Or, R a R b Together with the carbon atoms it is attached to, they form C 3-8 Cycloalkyl, 4-10 membered heterocyclic alkyl, with one or more R b-2 Replacement C 3-8 cycloalkyl, or by one or more R b-3 Substituted 4-10 membered heterocyclic alkyl groups;

[0175] R b-1 -OH, -CO2H, -CON(R) b-4 R b-5 ), -N(R b-6 )C(=O)R b-7C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 6-10 aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclic alkyl, C substituted with one or more hydroxyl groups 1-6 Alkyl, with one or more R b-8 Replacement C 6-10 aryl, with one or more R b-9 Substituted 5-10 heteroaryl groups, or substituted with one or more R groups b-10 Substituted 4-10 membered heterocyclic alkyl groups;

[0176] R b-4 R b-5 and R b-6 Independently H or C 1-6 alkyl;

[0177] R b-7 C 1-6 Alkyl or 4-10 membered heterocyclic alkyl;

[0178] R b-2 R b-3 R b-8 R b-9 and R b-10 Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0179] p1 is 0, 1, 2, 3, 4, 5 or 6;

[0180] In the 4-10 membered heterocyclic alkyl and 5-10 membered heteroaryl groups, the heteroatom is one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3.

[0181] In some implementations, X4 is The carbonyl end is connected to X5;

[0182] R 4 -C(=O)CH3, -L 14 -Z 14 or -L 24 -(L 34 ) n1 -L 44 -Z 24 ;

[0183] n1 is 0, 1, or 2;

[0184] L 14 Same as L above 1 Definition of Z 14Same as Z above 1 Definition; L 24 Same as L above 2 Definition; L 34 Same as L above 3 Definition; L 44 Same as L above 4 Definition; Z 24 Same as Z above 2 Definition.

[0185] In some implementation schemes, R 4 -C(=O)R 4a ;R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl.

[0186] In some implementations, X4 is The carbonyl end is connected to X5;

[0187] R 4 -C(=O)R 4a , -L 14 -Z 14 or -L 24 -(L 34 ) n1 -L 44 -Z 24 ;

[0188] R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0189] n1 is 0, 1, or 2;

[0190] L 14 Same as L above 1 Definition of Z 14 Same as the definition of z1 above; L 24 Same as L above 2 Definition; L34 Same as L above 3 Definition; L 44 Same as L above 4 Definition; Z 24 Same as Z above 2 Definition.

[0191] In some implementations, L 14 For chemical bonds, -C(=O)-C 1-6 alkylene-C(=O)- or a-end and Z 14 Connection; R Lb and R Lc Independently H or C 1-6 alkyl;

[0192] Z 14 for Z 1b It is a halogen; Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0193] L 24 For chemical bonds, -C(=O)-C 1-6 alkylene-NH- b -C(=O)-C 1-6 Alkylene-C(=O)- b or The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 34 connect;

[0194] R Lb and R Lc Independently H or C 1-6 alkyl;

[0195] R Lf It is -NHC(=NH)NH2;

[0196] L 34 -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 44 connect;

[0197] L 44 for d end and Z end 24 connect;

[0198] Z 24 -C(=O)-C6-24 Alkylene -CO2H;

[0199] n1 is 0, 1, or 2;

[0200] n4, n5, n7, n8, n9, n10, and n11 are independently 1, 2, 3, 4, or 5;

[0201] m2, m3, m4, m6, m7, m8 and m9 are independently 1, 2, 3, 4 or 5.

[0202] In some implementations, L 14 It is -C(=O)-(CH2CH2O) m2a -C 1-6 Alkylene NH- a a end and Z end 1 Connections; m2a is 1-10;

[0203] Z 14 for

[0204] In some implementations, X4 is The carbonyl end is connected to X5.

[0205] In some implementations, X4 is The carbonyl end is connected to X5.

[0206] In some implementations, X4 is The carbonyl end is connected to X5.

[0207] In some implementations, X5 is The carbonyl end is connected to X6;

[0208] R 5a C 1-6 Alkyl, -Z 15 C that is replaced by -C(=O)NH2 1-6 alkyl;

[0209] Z 15 for Z 1d Z 1e and Z 1f Independently H or C 1-6 alkyl;

[0210] n8 can be 1, 2, 3, 4, or 5.

[0211] In some implementation schemes, R 5aFor -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 alkyl;

[0212] R 5a-1 and R 5a-2 Independently H or C 1-6 Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups;

[0213] R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 Cycloalkyl.

[0214] In some implementations, X5 is The carbonyl end is connected to X6;

[0215] R 5a C 1-6 Alkyl, -Z 15 Or be -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 Alkyl; R 5a-1 and R 5a-2 Independently H or C 1-6 Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups;

[0216] R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl;

[0217] Z 15 for Z 1d Z 1e and Z 1f Independently H or C 1-6 alkyl;

[0218] n8 can be 1, 2, 3, 4, or 5.

[0219] In some implementations, X5 is The carbonyl end is connected to X6.

[0220] In some implementations, X5 is The carbonyl end is connected to X6.

[0221] In some implementations, X6 is The carbonyl end is connected to X7; R a For H or C 1-6 Alkyl; R b -(CH2) p1 -R b- 1 ;R b-1 -OH, C 1-6 Alkyl groups, or C groups substituted with one or more hydroxyl groups 1-6 Alkyl group; p1 is 0 or 1.

[0222] In some implementations, X6 is The carbonyl end is connected to X7.

[0223] In some implementations, X6 is The carbonyl end is connected to X7.

[0224] In some implementations, X7 is The carbonyl end is connected to X8.

[0225] In some implementations, X8 is The carbonyl end is connected to X9.

[0226] In some implementation schemes, X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8.

[0227] In some implementations, X10 is The carbonyl end is connected to X11; ring B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or is bonded by one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups;

[0228] R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 It can be -NH2, -OH or halogen independently.

[0229] In some implementations, X10 is The carbonyl end is connected to X11; ring B is a 5-8 membered heterocyclic alkenyl group, or is connected by one or more R groups. 10-1Substituted 5-8 membered heterocyclic alkenyl groups;

[0230] R 10-1 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 It can be -NH2, -OH or halogen independently.

[0231] In some implementations, X10 is The carbonyl end is connected to X11;

[0232] R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy;

[0233] R 10a-1 and R 10a-2 Independently -NH2, -OH, or halogen;

[0234] R 10b and R 10c H and C independently 1-6 Alkyl, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen;

[0235] n2a is 0, 1, or 2;

[0236] n2b and n2 are independently 1 or 2.

[0237] In some implementations, X10 is The carbonyl end is connected to X11;

[0238] R 10a C replaced by -NH2 1-6 Alkoxy;

[0239] R 10b C 1-6 alkyl;

[0240] n2 is 1 or 2.

[0241] In some implementations, X10 is The carbonyl end is connected to X11; R 10b For H, or by one or more R 10b-1Replacement C 1-6 alkyl;

[0242] R 10c C 1-6 Alkyl, or with one or more R 10b-1 Replacement C 1-6 alkyl;

[0243] R 10b-1 Independently -NH2, -OH, or halogen;

[0244] n2a is 0, 1, or 2;

[0245] n2b and n2 are independently 1 or 2.

[0246] In some implementations, X10 is The carbonyl end is connected to X11;

[0247] R 10a C replaced by -NH2 1-6 Alkoxy;

[0248] R 10b and R 10c H and C independently 1-6 Alkyl, or with one or more R 10b-1 Replacement C 1-6 alkyl;

[0249] R 10b-1 Independently -NH2, -OH, or halogen;

[0250] n2a is 0, 1, or 2;

[0251] n2b and n2 are independently 1 or 2.

[0252] In some implementations, X10 is The carbonyl end is connected to X11.

[0253] In some implementations, X10 is The carbonyl end is connected to X11.

[0254] In some implementations, X10 is

[0255] In some implementations, X11 is The carbonyl end is connected to X12.

[0256] In some implementations, X12 is R a R bTogether with the carbon atoms it is attached to, they form C 3-8 Cycloalkyl, 4-10 membered heterocyclic alkyl, with one or more R b-2 Replacement C 3-8 cycloalkyl, or by one or more R b-3 Substituted 4-10 membered heterocyclic alkyl groups; R b-2 and R b-3 Independently halogen or C 1-6 alkyl.

[0257] In some implementations, X12 is The carbonyl end is connected to X13.

[0258] In some implementations, X12 is The carbonyl end is connected to X13.

[0259] In some implementations, X13 is The carbonyl end is connected to X14.

[0260] In some implementations, X14 is The carbonyl end is connected to X15.

[0261] In some implementations, X15 is The carbonyl end is connected to X16; T 1 and T 4 Independently -NH-, -NR 15 -、-O- or -S-; T 2 T 3 T 5 and T 6 Independent of N, CH or CR 15 ;R 15 The definition is as described above.

[0262] In some implementation schemes, R 15 It can be -NH2, -CH2COOH, -CH2CH2COOH or -CH2CONH2.

[0263] In some implementations, X15 is The carbonyl end is connected to X16.

[0264] In some implementations, X16 is

[0265] R 16 -NH2, -L 116 -Z 116 or -L 216 -(L 316 )n1 -L 416 -Z 216 ;

[0266] n1 is 0, 1, or 2;

[0267] L 116 Same as L above 1 Definition of Z 116 Same as Z above 1 Definition; L 216 Same as L above 2 Definition; L 316 Same as L above 3 Definition; L 416 Same as L above 4 Definition; Z 216 Same as Z above 2 Definition.

[0268] In some implementations, X16 is

[0269] R 16 -NH2, -L 116 -Z 116 or -L 216 -(L 316 ) n1 -L 416 -Z 216 ;

[0270] n1 is 0, 1, or 2;

[0271] L 116 Same as L above 1 Definition of Z 116 Same as Z above 1 Definition; L 216 Same as L above 2 Definition; L 316 Same as L above 3 Definition; L 416 Same as L above 4 Definition; Z 216 Same as Z above 2 Definition.

[0272] In some implementations, X16 is

[0273] R 16 -NH2, -L 116 -Z 116 or -L 216 -(L 316 ) n1 -L 416 -Z 216 ;

[0274] n1 is 0, 1, or 2;

[0275] L 116 Same as L above 1 Definition of Z 116 Same as Z above 1 Definition; L 216 Same as L above 2 Definition; L 316 Same as L above 3 Definition; L 416 Same as L above 4 Definition; Z 216 Same as Z above 2 Definition.

[0276] In some implementations, X16 is

[0277] R 16 -NH2, -L 116 -Z 116 or -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ;

[0278] n1 and n1a are independently 0, 1 or 2;

[0279] L 116 Same as L above 1 Definition of Z 116 Same as Z above 1 Definition; L 216 Same as L above 2 Definition; L 316 Same as L above 3 Definition; L 416 Same as L above 4 Definition; Z 216 Same as Z above 2 Definition.

[0280] In some implementations, L 116 It is a chemical bond;

[0281] Z 116 for Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0282] L 216 For chemical bonds, -NH-C1-6 alkylene-NH- b , -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 alkylene- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0283] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect;

[0284] L 416 Independently d end and Z end 216 connect;

[0285] Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H,

[0286] R L4 a and R L4 b is independently H or

[0287] Z 1g It is a halogen;

[0288] n1 is 0, 1, or 2;

[0289] n7, n8, n9, n9a, n9b, n10, n10a, n11, n12 and n13 are independently 1, 2, 3, 4 or 5;

[0290] m, m6, m7, m8, m9, m10, m11 and m12 are independently 1, 2, 3, 4 or 5.

[0291] In some implementations, L 216 -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf For -CONH2;

[0292] L 316 Independently -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect;

[0293] n9, n9a, and n9b are independently 1, 2, 3, 4, or 5;

[0294] L 416 Independently d end and Z end 216 Connection; n10a can be 1, 2, 3, 4, or 5 independently;

[0295] Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -SO3H,

[0296] In some implementations, L 116 It is a chemical bond;

[0297] Z 116 for Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0298] L 216 For chemical bonds, -NH-C 1-6 alkylene-NH- b , -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 alkylene- b or The C 1- 6-alkylene compounds are optionally R Lf Replacement; b-end and L 316 Connection; R Lf For -CONH2;

[0299] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect;

[0300] L 416 Independently d end and Z end 216 connect;

[0301] Z 216 -C(=O)-C 6-24 Alkylene -CO2H;

[0302] R L4a and R L4b Independently for H or

[0303] Z 1g It is a halogen;

[0304] n1 is 0, 1, or 2;

[0305] n7, n8, n9, n10, n11, n12, and n13 are independently 1, 2, 3, 4, or 5;

[0306] m, m6, m7, m8, m9, m10, m11 and m12 are independently 1, 2, 3, 4 or 5.

[0307] In some implementations, L 216 for b end and L 316 Connections; m13 is independently 0, 1, 2, 3, 4, or 5; m14, m15, and m16 are independently 1, 2, 3, 4, or 5; R Lb and R Lc The definition is as described above.

[0308] In some implementations, L 116 It is a chemical bond;

[0309] Z 116 for Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl;

[0310] L 216 For chemical bonds, -NH-C 1-6 alkylene-NH- b , -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 alkylene- b , The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf For -CONH2;

[0311] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect;

[0312] L 416 Independently d end and Z end 216 connect;

[0313] Z 216 -C(=O)-C 6-24 Alkylene -CO2H;

[0314] R L4a and R L4b Independently for H or

[0315] Z 1g It is a halogen;

[0316] n1 is 0, 1, or 2;

[0317] n7, n8, n9, n10, n11, n12, and n13 are independently 1, 2, 3, 4, or 5;

[0318] m, m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5;

[0319] m13 can be 0, 1, 2, 3, 4 or 5 independently.

[0320] In some implementations, X16 is

[0321] In some implementations, X16 is

[0322] In some implementations, X16 is

[0323] In some embodiments, the compound represented by formula (I) is a compound represented by formula (I-1):

[0324] The definitions of X4, X5, X8, X10, X15 and X16 are as described in any embodiment of the present invention.

[0325] In some embodiments, the compound represented by formula (I) is a compound represented by formula (I-2):

[0326] The definitions of X4, X5, X8, X10, X16 and ring A are as described in any embodiment of the present invention.

[0327] In some embodiments, the compound represented by formula (I) is a compound represented by formulas (I-3):

[0328] The definitions of X4, X5, X8, X15, X16 and ring B are as described in any embodiment of the present invention.

[0329] In some embodiments, the compound represented by formula (I) is a compound represented by formulas (I-4):

[0330] The definitions of X4, X5, X8, X16, ring A, and ring B are as described in any embodiment of the present invention.

[0331] In some embodiments, the compound represented by formula (I) is a compound represented by formulas (I-5):

[0332] The definitions of X4, X5, X6, X8, X10, X12, X15 and X16 are as described in any embodiment of the present invention.

[0333] In some embodiments, in the compound shown in formula (I-5), X4 is The carbonyl end is connected to X5;

[0334] X5 is The carbonyl end is connected to X6;

[0335] X6 is The carbonyl end is connected to an amino group;

[0336] X8 is The carbonyl end is connected to an amino group;

[0337] X12 is The carbonyl end is connected to an amino group;

[0338] X10 is The carbonyl terminus is connected to an amino group; ring B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or is bonded to one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups;

[0339] R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2;

[0340] X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0341] X16 is

[0342] R 16 -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ;

[0343] L 216 -NH-C 1-6 alkylene-NH- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0344] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=o)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect;

[0345] L 416 Independently d end and Z end 216 connect;

[0346] Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H,

[0347] n1 and n1a are independently 0, 1 or 2;

[0348] n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5;

[0349] m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5;

[0350] m13 can be 0, 1, 2, 3, 4 or 5 independently;

[0351] In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is one, two or three.

[0352] In some embodiments, in the compound shown in formula (I-5), X10 is The carbonyl end is connected to X11.

[0353] In some embodiments, in the compound shown as formula (I-5), X15 is The carbonyl end is connected to X16; preferably In some embodiments, in the compounds shown as formula (I-5), X16 is...

[0354] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ia):

[0355] Among them, X5, X8, X16, and R 4 T 1 T 2 and T 3 The definition is as described in any embodiment of this invention.

[0356] In some embodiments, in the compound represented by formula (Ia), R4 -C(=O)CH3,

[0357] X5 is The carbonyl end is connected to an amino group;

[0358] X8 is The carbonyl end is connected to an amino group;

[0359] T 1 -NH-, -NR 15 -、-O- or -S-; T 2 and T 3 Independent of N, CH or CR 15 ;R 15 It can be -NH2, -CH2COOH, -CH2CH2COOH or -CH2CONH2;

[0360] X16 is defined as described in any embodiment of this invention.

[0361] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ib):

[0362] Among them, X5, X16, R 4 R 10b The definitions of n2 and n2 are as described in any embodiment of the present invention.

[0363] In some embodiments, in the compound represented by formula (Ib), R 10b C 1-6 Alkyl group; n2 is 1 or 2;

[0364] X5 is The carbonyl end is connected to an amino group;

[0365] R 4 -C(=O)CH3,

[0366] X16 is R 16 -L 216 -(L 316 ) n1 -L 416 -Z 216 ;

[0367] L 216 -NH-C 1-6 alkylene-NH- b , The C 1-6Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0368] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect;

[0369] L 416 Independently d end and Z end 216 connect;

[0370] Z 216 -C(=O)-C 6-24 Alkylene -CO2H;

[0371] n1 is 0, 1, or 2;

[0372] n9 and n10 are independently 1, 2, 3, 4 or 5;

[0373] m6, m7, m8 and m9 are independently 1, 2, 3, 4 or 5.

[0374] In some embodiments, in the compound represented by formula (Ib), X16 is

[0375] In some embodiments, in the compound represented by formula (Ib), R 10b C 1-6 Alkyl group; n2 is 1 or 2;

[0376] X5 is The carbonyl end is connected to the amino group above it;

[0377] X16 is

[0378] X4 is The carbonyl end is connected to X5;

[0379] R 4 -L 24 -(L 34 ) n1 -L 44 -Z 24 ;

[0380] L 24 For chemical bonds, b end and L 34 connect;

[0381] L 34 -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 44 connect;

[0382] L 44 for d end and Z end 24 connect;

[0383] Z 24 -C(=O)-C 6-24 Alkylene -CO2H;

[0384] n1 is 0, 1, or 2;

[0385] n9 and n10 are independently 1, 2, 3, 4 or 5.

[0386] In some embodiments, in the compound shown in formula (Ib), X4 is

[0387] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ic):

[0388] Among them, X5, X16, R 4 T 1 T 2 T 3 R 10b The definitions of n2 and n2 are as described in any embodiment of the present invention.

[0389] In some embodiments, the compound shown in formula (I) is a compound shown in formula (Id):

[0390] Among them, X5, X8, X16, and R 4 T 4 T 5 and T 6 The definition is as described in any embodiment of this invention.

[0391] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ie):

[0392] Among them, X5, X10, X16 and R 4 The definition is as described in any embodiment of this invention.

[0393] In some embodiments, in the compound represented by formula (Ie), X10 is

[0394] X5 is

[0395] R 4 -C(=O)CH3,

[0396] X16 is R 16 -L 216 -(L 316 ) n1 -L 416 -Z 216 ;

[0397] L 216 -NH-C 1-6 alkylene-NH- b or The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0398] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect;

[0399] L 416 Independently d end and Z end 216 connect;

[0400] Z 216 -C(=O)-C 6-24 Alkylene -CO2H;

[0401] n1 is 0, 1, or 2;

[0402] n9 and n10 are independently 1, 2 or 3;

[0403] m6 and m7 are independently 1, 2, 3, 4 or 5.

[0404] In some embodiments, in the compound represented by formula (Ie), X16 is

[0405] In some embodiments, the compound represented by formula (I) is a compound represented by formula (If):

[0406] Among them, X5, X8, X15, X16, and R 10c n2a, n2b and R 4 The definition is as described in any embodiment of this invention.

[0407] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ig):

[0408] Among them, R 4 -C(=O)CH3,

[0409] X5 is The carbonyl end is connected to an X6 amino group;

[0410] X6 is The carbonyl end is connected to an amino group;

[0411] X8 is The carbonyl end is connected to an amino group;

[0412] X12 is The carbonyl end is connected to an amino group;

[0413] X10 is The carbonyl end is connected to an amino group;

[0414] Cycle B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or a ring with one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups;

[0415] R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2;

[0416] X15 is The carbonyl end is connected to X16;

[0417] Ring A is a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0418] X16 is

[0419] R 16 -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ;

[0420] L 216 -NH-C 1-6 alkylene-NH- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0421] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect;

[0422] L416 Independently d end and Z end 216 connect;

[0423] Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H,

[0424] n1 and n1a are independently 0, 1 or 2;

[0425] n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5;

[0426] m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5;

[0427] m13 can be 0, 1, 2, 3, 4 or 5 independently;

[0428] In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is one, two or three.

[0429] In some embodiments, in the compound shown in formula (Ig), X10 is... The carbonyl end is connected to an amino group.

[0430] In some embodiments, in the compound shown in formula (Ig), X15 is... The carbonyl end is connected to an X16 amino group.

[0431] In some embodiments, the compound represented by formula (I) is a compound represented by formula (Ih):

[0432] Wherein, ring B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cyclic alkenyl group, or is surrounded by one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups;

[0433] R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R10b-1 Independently -NH2;

[0434] Ring A is a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl;

[0435] X16 is

[0436] R 16 -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ;

[0437] L 216 -NH-C1-6 alkylene-NH- b , or -NH-C1-6 alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl;

[0438] L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a-NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect;

[0439] L 416 Independently d end and Z end 216 connect;

[0440] Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H,

[0441] n1 and n1a are independently 0, 1 or 2;

[0442] n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5;

[0443] m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5;

[0444] m13 can be 0, 1, 2, 3, 4 or 5 independently;

[0445] In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is one, two or three.

[0446] In some embodiments, the compound as shown in formula (Ih), for

[0447] In some embodiments, the compound as shown in formula (Ih), for

[0448] In some embodiments, the compound represented by formula (I) is any one of the compounds in Table A below:

[0449] Table A

[0450] In some embodiments, the compound represented by formula (I) is any one of the compounds in Table B below:

[0451] Table B

[0452] In some embodiments, when the compounds in Tables A and B contain cationic fragments, the anion is acetate.

[0453] The present invention provides a pharmaceutical composition comprising a compound as shown in formula (I) as described above or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

[0454] The present invention also provides the use of a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for the prevention and / or treatment of diseases associated with interleukin-23; wherein the diseases associated with interleukin-23 are preferably autoimmune diseases, such as psoriasis, arthritis or inflammatory bowel disease.

[0455] The present invention also provides a compound as shown in formula (II) or a pharmaceutically acceptable salt thereof.

[0456] Among them, R 4 -C(=O)R 4a ;R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl;

[0457] X16 is R 16 It is -NH2;

[0458] X5, X6, X8, X10, and X12 are defined as in any embodiment of the present invention.

[0459] In some embodiments, the compound represented by formula (II) is any of the following compounds:

[0460] The present invention also provides compounds or pharmaceutically acceptable salts thereof as shown below:

[0461] Terminology Definition

[0462] Unless otherwise specified, the terms used in this invention have the following meanings:

[0463] The amino acid residues in this invention include natural amino acid residues and non-natural amino acid residues; the natural amino acids refer to 20 conventional amino acids, namely alanine (A), cysteine ​​(C), aspartic acid (D), glutamic acid (E), phenylalanine (F), glycine (G), histidine (H), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), proline (P), glutamine (Q), arginine (R), serine (S), threonine (T), valine (V), tryptophan (W), and tyrosine (Y);

[0464] The non-natural amino acids mentioned refer to amino acids that are not naturally encoded or found in the genetic code of any organism; they can be purely synthetic compounds. Furthermore, in this invention, the amino acid residues also include natural or non-natural amino acids whose C-terminal carboxyl group, N-terminal amino group, and / or their side-chain functional groups are chemically modified.

[0465] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0466] The term "alkyl" refers to a straight-chain or branched alkyl group having a specified number of carbon atoms (e.g., C1 to C6). Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, etc.

[0467] The term "halogenated alkyl" refers to an alkyl group that is substituted with one or more halogens, such as an alkyl group substituted with one, two, three or four halogens.

[0468] The term "alkoxy" refers to the group R. X -O-, where R X It is an alkyl group as defined above.

[0469] In this invention, the term "alkenyl" refers to a straight-chain or branched hydrocarbon chain group having at least one double bond, consisting only of carbon and hydrogen atoms, having, for example, 2 to 6 (preferably 2 to 4) carbon atoms, and connected to the rest of the molecule by single bonds, such as including but not limited to vinyl, 1-propenyl, n-allyl, but-1-enyl, but-2-enyl, pent-1-enyl, or pent-1,4-dienyl.

[0470] The term "alkynyl" refers to a straight-chain or branched hydrocarbon chain group having at least one triple bond, consisting only of carbon and hydrogen atoms, having, for example, 2 to 6 (preferably 2 to 4) carbon atoms, and connected to the rest of the molecule by single bonds, such as including but not limited to ethynyl, 1-propynyl, n-propynyl, but-1-alkynyl, but-2-alkynyl, pent-1-alkynyl, or pent-1,4-dialkynyl.

[0471] The term "oxo" refers to =O, where an oxygen atom replaces two hydrogen atoms on the same atom. For example, methylene (-(CH2-)) becomes carbonyl (-C(=O)-) after being oxidized.

[0472] The term "cycloalkyl" refers to a saturated monocyclic cyclic group consisting only of carbon atoms and having a specified number of carbon atoms (e.g., C3 to C8). Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0473] The term "cycloalkenyl" refers to a group having at least one carbon-carbon sp. 2 Non-aromatic carbocyclic substituents (e.g., C7-C) of partially unsaturated monocyclic or polycyclic (e.g., bicyclic, tricyclic or more bridging rings, fused rings, or spirocyclic systems) of double bonds. 15 Cycloalkenyl, C7-C 10 Cycloalkenyl, C8-C9 cycloalkenyl, C3-C 12 Cycloalkenyl, C3-C 10 Cycloalkenyl or C5-C6 cycloalkenyl. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, and cyclohexenyl.

[0474] In this invention, the term "heterocyclic alkenyl" refers to a cyclic, unsaturated group having a specified number of ring atoms (e.g., 3-10, 3-6, or 8-10), a specified number of heteroatoms (e.g., 1, 2, 3, 4, or 5), and a specified type of heteroatom (one, two, or more of N, O, and S); and having one or more (e.g., 1, 2, or 3) sp... 2 Double bonds, which are monocyclic or polycyclic (e.g., bridged rings, fused rings, or spirocyclic systems of bicyclic, tricyclic, or more rings), are not aromatic. Heterocyclic alkenyl groups are attached to the rest of the molecule via carbon atoms or heteroatoms.

[0475] The term "heterocyclic alkyl" refers to a cyclic group having a specified number of ring atoms (e.g., 5 to 10), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S), which can be monocyclic, bridged, or spirocyclic, and each ring is saturated. Heterocyclic alkyl groups include, but are not limited to, aza-butanediol, tetrahydropyrrolyl, tetrahydrofuranyl, morpholinyl, and piperidinyl.

[0476] The term "aryl" refers to an aryl group having a specified number of carbon atoms (e.g., C6 to C5). 10 An aryl group is a cyclic group consisting solely of carbon atoms, which may be monocyclic or polycyclic, and at least one ring is aromatic (conforming to Hückel's rule). An aryl group is linked to other segments of the molecule via an aromatic or non-aromatic ring. Aryl groups include, but are not limited to, phenyl and naphthyl groups.

[0477] The term "heteroaryl" refers to a cyclic group having a specified number of ring atoms (e.g., 5 to 10), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (one or more of N, O, and S). It can be monocyclic or polycyclic, and at least one ring is aromatic (conforming to Hückel's rule). Heteroaryl groups are linked to other segments of a molecule via aromatic or non-aromatic rings. Heteroaryl groups include, but are not limited to, furanyl, pyrroleyl, thiopheneyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, and indoleyl groups.

[0478] As used in this invention, the terms "cycloalkylene", "heterocycloalkylene", "arylene", and "heteroaryl" refer to a cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group in which one hydrogen atom is further substituted, as defined above.

[0479] The "-" at the end of a group indicates that the group is connected to other segments in the molecule through that site. For example, CH3-C(=O)- refers to an acetyl group.

[0480] In structural fragments This refers to the connection between this structural segment and other segments in the molecule through this site. Unless otherwise specified, use wedge-shaped solid lines. and wedge-shaped dashed key This indicates the absolute configuration of a stereocenter. When the compound is chiral, it is either the R configuration or the S configuration.

[0481] The term "multiple" refers to 2, 3, 4, or 5.

[0482] When any variable (e.g., group R) 10a-4 When these terms appear multiple times in the definition of a compound, their definitions are independent and do not affect each other. For example, a compound defined by three R's... 10a-4 Replacement C 1-6 Alkyl refers to C 1-6 Alkyl groups will be formed by 3 Rs 10a-4 Replace, 3 R 10a-4 The definitions are independent of each other and do not affect each other.

[0483] The term "pharmaceutically acceptable salt" refers to a salt obtained by reacting a compound with a pharmaceutically acceptable (relatively non-toxic, safe, and suitable for patient use) acid or base. When a compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable base in a suitable inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, sodium, potassium, calcium, aluminum, magnesium, bismuth, and ammonium salts. When a compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the free form of the compound with a sufficient amount of a pharmaceutically acceptable acid in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include, but are not limited to, hydrochloride salts, sulfate salts, and methanesulfonate salts.

[0484] The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the preparation of prescriptions. They are all substances contained in pharmaceutical preparations, excluding the active ingredient.

[0485] The term “treatment” refers to any of the following: (1) alleviating one or more biological manifestations of a disease; (2) interfering with one or more points in the biological cascade that triggers the disease; or (3) slowing the development of one or more biological manifestations of a disease.

[0486] The term "prevention" refers to reducing the risk of developing a disease.

[0487] The term "patient" refers to any animal that has received or is about to receive treatment, preferably a mammal, and most preferably a human. Mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans.

[0488] Based on common knowledge in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0489] The reagents and raw materials used in this invention are all commercially available.

[0490] The positive and progressive effects of this invention are as follows: This invention provides a polypeptide compound that specifically blocks the IL-23 signaling pathway, and the polypeptide compound provided by this invention has at least one of the following advantages:

[0491] (1) Extremely strong biological activity;

[0492] (2) Good pharmacokinetic properties;

[0493] (3) It showed good efficacy in animal medicine. Detailed Implementation

[0494] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0495] This application uses the following abbreviations:

[0496] DCM represents dichloromethane; DCE represents dichloroethane; PE represents petroleum ether; Dioxane or 1,4-dioxane represents dioxane; DMF represents dimethylformamide; DMSO represents dimethyl sulfoxide; EA or EtOAc represents ethyl acetate; MeCN or ACN represents acetonitrile; MeOH represents methanol; MTBE represents methyl tert-butyl ether; DMA or DMAC represents N,N-dimethylacetamide; H2O represents water.

[0497] Pip represents piperidine; HATU represents (7-azobenzotriazole)-N,N,N′,N′-tetramethylurea hexafluorophosphate; HOAT represents N-hydroxy-7-azabenzotriazole; HOBT represents 1-hydroxybenzotriazole; DIPEA or DIEA represents N,N-diisopropylethylamine; TEA represents triethylamine; TBAF represents tetrabutylammonium fluoride; MsCl represents methanesulfonyl chloride; NaH represents sodium hydride; NaBH4 represents sodium borohydride; Na2CO3 represents sodium carbonate; NaCl represents sodium chloride; NaHCO3 represents sodium bicarbonate; NaOH represents hydroxide. Sodium sulfate; Na2SO4 represents sodium sulfate; NBS represents N-bromosuccinimide; K2CO3 represents potassium carbonate; Cs2CO3 represents cesium carbonate; AcOH represents acetic acid; Ac2O represents acetic anhydride; HCl represents hydrochloric acid; TFA represents trifluoroacetic acid; Tis represents triisopropylsilane; EDT represents 1,2-ethylenedithiol; NH4I represents ammonium iodide; I2 represents elemental iodine; Oxyma represents ethyl 2-oxime cyanoacetate; DIC represents N,N′-diisopropylcarbodiimide; PhSiH3 represents benzenesilane; 1,3-Dimethylbarbituric acid acid represents 1,3-dimethylbarbituric acid; Pd(PPh4)3 represents tetrakis(triphenylphosphine)palladium; DBU represents 1,8-diazacyclic [5,4,0]undecene-7; DME·NiCl2 represents nickel chloride dimethoxyethane; Mn represents manganese powder; Zn represents zinc powder; dtbpy represents 4,4′-di-tert-butyl-2,2′-bipyridine; NiBr represents nickel bromide; TPP represents triphenylphosphine; (2,2'-Bipyridine) NiCl2 represents (2,2'-bipyridine)nickel dichloride; DEA represents diethylamine; TSTU represents 2-succinimide-1,1,3,3-tetramethylurea tetrafluoroborate; SEMCl represents 2-(trimethylsilyl)ethoxymethyl chloride; Alloc-Osu represents allyl succinimide carbonate; Fmoc-OSu represents 9-fluorenylmethyl N-succinimide carbonate; MeI represents iodomethane; CDI represents N,N′-carbonyldiimidazole;

[0498] Fmoc represents 9-fluorenylmethoxycarbonyl; Trt represents triphenylmethyl; Boc represents tert-butyloxycarbonyl; t Bu represents tert-butyl; Ac represents acetyl; Me represents methyl; All represents allyl; Dde represents 1-(4,4-dimethyl-2,6-dioxocyclohexyl-1-ethylene)ethyl;

[0499] h represents hours; min represents minutes; mg represents milligrams; mL represents milliliters; M represents mol; mmol represents millimoles; g represents grams; v or V represents volume; %wt represents weight ratio; N2 represents nitrogen; HPLC represents high performance liquid chromatography; d represents doublet; br represents broad peak; m represents multiplet; ℃ represents degrees Celsius; CD3OD represents deuterated methanol; CDCl3 represents deuterated chloroform; DMSO-d6 represents deuterated dimethyl sulfoxide; LC-MS represents liquid chromatography-mass spectrometry; ESI represents electrospray ionization; m / z represents mass-to-charge ratio.

[0500] Amino acid list:

[0501] Segment list

[0502] Example 1: Intermediate Synthesis:

[0503] Intermediate AA1

[0504] Compound AA1-1 (5 g, 25.3 mmol) was dissolved in DMF (50 mL). Sodium hydride (1.2 g, 30.2 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 0.5 h. SEMCl (4.13 g, 27.7 mmol) was added at 0 °C. The mixture was stirred at room temperature (25 °C) for 1 h. Water (1500 mL) was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 30%) to obtain compound AA1-2. MS m / z (ESI): 328.3 [M+H] + .

[0505] Activated zinc powder (0.7 g, 10.6 mmol) was added to DMF (5 mL), and iodine (0.077 g, 0.3 mmol) was added under nitrogen atmosphere. The system was stirred at room temperature (25 °C) for 0.5 h. AA1-4 (2.06 g, 4.6 mmol) in DMF (10 mL) was added to the system; the system was stirred at room temperature (25 °C) for 0.5 h, and the supernatant A was collected. Under nitrogen atmosphere, AA1-2 (1 g, 3 mmol), tris(dibenzylacetone)palladium (0.28 g, 0.3 mmol), and tricyclohexylphosphine (0.17 g, 0.6 mmol) were added to DMF (10 mL), and supernatant A was added; the system was heated to 80 °C and stirred for 4 h. Water (200 mL) was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 25%) to give compound AA1-3. MS m / z (ESI): 573.5 [M+H] + .

[0506] Compound AA1-3 (1.5 g, 2.6 mmol) was dissolved in THF (12 mL) and H2O (3 mL). Lithium hydroxide (0.15 g, 5.2 mmol) was added at 0 °C. The mixture was stirred for 1 hour in an ice bath. Water (20 mL) was added to the system, and the system was adjusted to neutral with 1 mol / L hydrochloric acid aqueous solution. The mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to obtain compound AA1. MS m / z (ESI): 559.5 [M+H] + .

[0507] Starting from the corresponding starting materials, intermediates AA6, AA7, AA8, AA9, AA21, AA22, and AA24 were synthesized by referring to the operation steps of intermediate AA1.

[0508] AA6: MS m / z(ESI): 558.6[M+H] + . 1H-NMR(400MHz,DMSO-d6)δ9.31(s,1H),8.38-8.31(m,2H),7.93(q,J=7.6Hz,3H),7.64(d,J=7.2Hz,2H),7.49-7.27(m,4H),7.10(d,J=2.8Hz,1H),5.84(s,2H),4.52-4.46(m,1H),4.27-4.11(m,3H),3.60-3.49(m,4H),0.89-0.80(m,2H),-0.07(s,9H).

[0509] AA7:MS m / z(ESI):558.6[M+H] + 。 1 H-NMR(400MHz,DMSO-d6)δ9.17(s,1H),8.24-8.15(m,2H),7.87(d,J=7.6Hz,3H),7.57(d,J=7.6Hz,2H),7.41-7.25(m,4H),6.97(d,J=2.8Hz,1H),5.75(s,2H),4.43-4.37(m,1H),4.18-4.09(m,3H),3.47-3.41(m,4H),0.81-0.71(m,2H),-0.15(s,9H).

[0510] AA8:MS m / z(ESI):559.4[M+H] + 。

[0511] AA9:MS m / z(ESI):543.4[M+H] + 。 1 H-NMR(400MHz,DMSO-d6)δ8.79(s,1H),8.17(s,1H),8.01(s,1H),7.77-7.69(m,2H),7.52(t,J=5.6Hz,2H),7.28-7.15(m,4H),5.63-5.47(m,2H),4.13-3.97(m,3H),3.79-3.69(m,1H),3.04(s,1H),2.93-2.88(m,1H),1.23(s,9H).

[0512] AA21:MS m / z(ESI):486.2[M+H] + 。

[0513] AA22:MS m / z(ESI):557.2[M+H] + 。

[0514] AA24: MS m / z(ESI): 674.2[M+H] + .

[0515] Intermediate AA2

[0516] Compound AA2-1 (5 g, 22.12 mmol) was dissolved in DMAC (70 mL). NaH (1.06 g, 26.54 mmol, 60% purity) was added at 0 °C, and the mixture was stirred at 0 °C for 0.5 h. MeI (4.71 g, 33.18 mmol) was added dropwise at 0 °C. The mixture was then heated to room temperature (25 °C) and stirred for 2 h. Water was added to the mixture, and the pH was adjusted to 6 with 2N dilute hydrochloric acid. The mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum (v / v) = 25%) to obtain compound AA2-2. MS m / z (ESI): 239.9 [M+H] + .

[0517] Compound AA2-2 (4.35 g, 18.12 mmol) was dissolved in DMA (50 mL), and DME·NiCl2 (398.08 mg, 1.81 mmol), Mn (4.98 g, 90.59 mmol), compound AA1-4 (12.26 g, 27.18 mmol), and pyridine-2-formamidinium (856.61 mg, 5.44 mmol) were added. The system was heated to 60 °C and stirred for 2 h under N2 conditions. The system was then poured into ice water and stirred until homogeneous, followed by extraction three times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 10%) to obtain compound AA2-3. MS m / z (ESI): 485.3 [M+H] + .

[0518] Compound AA2-3 (2 g, 4.13 mmol) was dissolved in DCE (20 mL), and trimethyltin hydroxide (2.99 g, 16.51 mmol) was added. The system was heated to 65 °C and stirred for 12 h under N2 conditions. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 10%) to give compound AA2. MS m / z (ESI): 471.1 [M+H] + . 1H-NMR (400MHz, DMSO-d6) δ7.86 (d, J=7.4Hz, 2H), 7.69 (d, J=7.7Hz, 1H), 7.59-7.47 (m, 2H), 7.38 (t, J=7.4Hz, 2H), 7.26 (q, J=7.1Hz, 2H), 7.15 (d , J=8.0Hz, 1H), 7.04 (s, 1H), 6.89-6.84 (m, 1H), 4.28-4.24 (m, 1H), 4.14 -4.03(m, 3H), 3.18-3.12(m, 1H), 3.02-2.87(m, 5H), 2.79-2.66(m, 3H).

[0519] Starting from the corresponding starting materials, intermediates AA11 and AA12 were synthesized by referring to the operation steps of intermediate AA2.

[0520] AA11: MS m / z(ESI): 443.1[M+H] + .

[0521] AA12: MS m / z(ESI): 443.2[M+H] + . 1 H-NMR (400MHz, DMSO-d6) δ8.40 (s, 1H), 7.88 (d, J=7.6Hz, 2H), 7.64-7.49 (m, 3H), 7.42-7.26(m, 6H), 6.92(brs, 1H), 4.37-3.99(m, 6H), 3.19-2.94(m, 2H).

[0522] Intermediate AA3

[0523] NiBr (892.42 mg, 3.77 mmol) and dtbpy (1.01 g, 3.77 mmol) were dissolved in DMA (30 mL). The system was stirred at room temperature (25 °C) for 1 h under N2 conditions. Compounds AA3-1 (4.87 g, 21.56 mmol), AA3-2 (6 g, 10.78 mmol), and Zn (3.17 g, 48.51 mmol) were added. The system was heated to 40 °C and stirred for 2 h under N2 conditions. Water was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 50%) to obtain compound AA3-3. MS m / z (ESI): 513.4 [M+H] + .

[0524] Compound AA3-3 (1 g, 1.95 mmol) was dissolved in DCM (8 mL), and TFA (8 mL) was added. The system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 50%) to give compound AA3. MS m / z (ESI): 457.2 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ12.83 (s, 1H), 7.89-7.78 (1n, 3H), 7.64-7.55 (m, 3H) , 7.45-7.37(m, 4H), 7.31-7.24(m, 2H), 4.41-4.13(m, 6H), 3.22-2.94(m, 5H).

[0525] Intermediate AA13

[0526] Compound AA13-1 (10 g, 52.85 mmol) was dissolved in DMF (100 mL), and potassium carbonate (14.61 g, 105.70 mmol) and allyl bromide (8.31 g, 68.71 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h. Water (200 mL) was added to the system, and the mixture was extracted with ethyl acetate (200 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 5%) to obtain compound AA13-2. MS m / z (ESI): 252.2 [M + Na] + .

[0527] Compound AA13-2 (11 g, 47.98 mmol) was dissolved in 120 mL of dioxane hydrochloride solution; the system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to give compound AA13-3, which was used directly in the next reaction without further purification. MS m / z (ESI): 130.3 [M+1] + .

[0528] Compound AA1 (20 g, 35.80 mmol) was dissolved in DMF (100 mL), and potassium carbonate (9.9 g, 71.60 mmol) and allyl bromide (5.64 g, 46.54 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h. Water (300 mL) was added to the system, and the mixture was extracted with ethyl acetate (300 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 30%) to give compound AA13-4. MS m / z (ESI): 599.6 [M + H] + .

[0529] Compound AA13-4 (13 g, 21.71 mmol) and DEA (6.67 g, 1.19 mol, 75 mL) were dissolved in ACN (75 mL); the system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (petroleum ether / tetrahydrofuran (v / v) = 33%) to obtain compound AA13-5.

[0530] Compound AA13-5 (7.75 g, 20.58 mmol) and Fmoc-Asp (O t Bu)-OH (8.47 g, 20.58 mmol) was dissolved in DMF (100 mL), and HATU (11.73 g, 30.87 mmol) and DIPEA (7.98 g, 61.75 mmol) were added. The system was stirred at room temperature (25 °C) for 1 h. Water (300 mL) was added to the system, and the mixture was extracted with ethyl acetate (300 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 35%) to give compound AA13-6. MS m / z (ESI): 770.6 [M+H] + .

[0531] Compound AA13-6 (14 g, 18.18 mmol) and tetraphenylphosphine palladium (1.68 g, 1.45 mmol) were dissolved in dichloromethane (150 mL). Under nitrogen atmosphere, phenylsilane (15.74 g, 145.46 mmol) was added. The system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 65%) to give compound AA13-7. MS m / z (ESI): 730.4 [M+H] + .

[0532] Compounds AA13-7 (14.00 g, 14.39 mmol) and AA13-3 (2.85 g, 17.28 mmol) were dissolved in DMF (100 mL), and HATU (8.20 g, 21.58 mmol) and DIPEA (5.57 g, 43.2 mmol) were added. The system was stirred at room temperature (25 °C) for 1 h. Water (300 mL) was added to the system, and the mixture was extracted with ethyl acetate (300 mL × 3). The organic phases were combined, washed with saturated brine (300 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 30%) to obtain compound AA13-8. MS m / z (ESI): 841.6 [M+H] + .

[0533] Compound AA13-8 (10.00 g, 10.30 mmol) was dissolved in dichloromethane (150 mL), and zinc bromide (18.54 g, 82.40 mmol) was added. The system was stirred at room temperature (25 °C) for 1 h. The mixture was quenched with an aqueous sodium bicarbonate solution (500 mL), filtered, and the aqueous phase was extracted with dichloromethane (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by C18 reversed-phase medium-pressure column chromatography (water:acetonitrile = 3:2) to obtain intermediate AA13. MS m / z (ESI): 785.7 [M+H] + .

[0534] Intermediate AA14

[0535] Activated zinc powder (1.5 g, 23.3 mmol) was added to DMF (10 mL), and iodine (0.12 g, 0.46 mmol) was added under a nitrogen atmosphere; the system was stirred at 0 °C for 0.5 h. A DMF (10 mL) solution of Fmoc-3-iodo-L-alanine methyl ester (3.15 g, 7.0 mmol, 1.5 equivalents) was added; the system was stirred at room temperature (25 °C) for 0.5 h. Under a nitrogen atmosphere, the supernatant was added to a DMF (10 mL) solution of compound AA14-1 (1 g, 4.7 mmol), tris(dibenzylideneacetone)dipalladium (0.43 g, 0.46 mmol), and 2-dicyclohexylphosphine-2′,6′-dimethoxybiphenyl (0.38 g, 0.93 mmol); the system was stirred at 80 °C for 16 h. Water (100 mL) was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (petroleum ether / ethyl acetate (v / v) = 25%) to give compound AA14-2. MS m / z (ESI): 460.2 [M+H] + .

[0536] Compound AA14-2 (1.2 g, 2.6 mmol) and trimethyltin hydroxide (1.4 g, 7.8 mmol) were dissolved in DCE (20 mL); the system was stirred at 80 °C for 2 h. Dichloromethane (20 mL) was added to the system, the organic phase was washed with saturated citric acid solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound AA14, which was used directly in the next reaction without further purification. MS m / z (ESI): 446.2 [M+H] + .

[0537] Starting from the corresponding starting materials, intermediates AA15, AA16, AA17, AA18, AA19, and AA25 were synthesized by referring to the operation steps of intermediate AA14.

[0538] AA15: MS m / z(ESI): 529.2[M+H] + .

[0539] AA16: MS m / z(ESI): 430.3[M+H] + .

[0540] AA17: MS m / z(ESI): 428.2[M+H] + .

[0541] AA18: MS m / z(ESI): 430.2[M+H] + .

[0542] AA19: MS m / z(ESI): 489.2[M+H- t Bu] + .

[0543] AA25: MS m / z(ESI): 443.2[M+H] + .

[0544] Intermediate AA23

[0545] Compound AA23-1 (10 g, 24.30 mmol) was dissolved in DMF (100 mL), and HATU (13.85 g, 36.46 mmol), DIEA (9.42 g, 72.91 mmol), and pyrrolidine (1.73 g, 24.30 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h. Water (500 mL) was added, and the mixture was extracted three times with ethyl acetate (300 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 0–50%) to give compound AA23-2. MS m / z (ESI): 465.3 [M+H] + .

[0546] Compound AA23-2 (10 g, 21.53 mmol) was dissolved in DCM (50 mL), and TFA (50 mL) was added. The system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to obtain a crude product, which was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water = 0–50%) to obtain compound AA23. MS m / z (ESI): 409.3 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ7.90 (d, J=7.6Hz, 2H), 7.70 (d, J=7.6Hz, 2H), 7.44-7.31 (m, 4H), 6.79 (d, J=7.2H z, 1H), 4.26-4.22 (m, 3H), 4.06-4.01 (m, 1H), 3.25 (t, J=6.4Hz, 4H), 2.64-2.63 (m, 2H), 1.85-1.71 (m, 4H).

[0547] Fmoc-Asp-Pal-Sar-OAll

[0548] Fmoc-3-Pal-OH (4 g, 10.30 mmol) was dissolved in DMF (50 mL), and sarcosine tert-butyl hydrochloride (2.24 g, 12.36 mmol), HATU (5.90 g, 15.45 mmol), and DIEA (3.99 g, 30.89 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h. Water (50 mL) was added to the system, and the mixture was extracted with ethyl acetate (50 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 0–40%) to give compound BB1-1. MS m / z (ESI): 515.9 [M+H] + .

[0549] Compound BB1-1 (5 g, 9.70 mmol) was dissolved in DCM (15 mL), and TFA (15 mL) was added. The system was stirred at room temperature (25 °C) for 16 h. The system was concentrated to obtain a crude product, which was purified by C18 reversed-phase medium-pressure column chromatography (water / acetonitrile = 50%) to give compound BB1-2. MS m / z (ESI): 459.9 [M+1] + .

[0550] Compound BB1-2 (4 g, 8.71 mmol) was dissolved in DEA (20 mL) and ACN (20 mL). The system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to give compound BB1-3, which was used directly in the next reaction without further purification. MS m / z (ESI): 238.2 [M+H] + .

[0551] Fmoc-Asp( t Bu)-OH (3.5 g, 8.51 mmol) was dissolved in THF (30 mL), and DIEA (2.20 g, 17.01 mmol) and TSTU (2.56 g, 8.51 mmol) were added. The system was stirred at room temperature (25 °C) for 16 h. Water (30 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound BB1-4, which was used directly in the next reaction without further purification. MS m / z (ESI): 530.8 [M + Na] + .

[0552] Compound BB1-3 (2 g, 8.43 mmol) was dissolved in THF (30 mL), and compound BB1-4 (4.29 g, 8.43 mmol) and DIEA (2.18 g, 16.86 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h. Water (30 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 0–20%) to obtain compound BB1-5. MS m / z (ESI): 630.9 [M+H] + . 1 H-NMR (400MHz, DMSO-d6) δ11.75 (s, 1H), 8.40-8.30 (m, 2H), 7.93-8.02 (1H), 7.8 3-7.91 (m, 2H), 7.68 (t, J = 8.0Hz, 2H), 7.64-7.57 (m, 2H), 7.40 (t, J = 7.4Hz, 2H), 7.30(t, J=7.1Hz, 2H), 7.19-7.13(m, 1H), 4.95-4.69(m, 1H), 4.34-4.11(m, 4H), 3.82-3.68(m, 2H), 3.03-2.75(m, 5H), 1.85-1.63(m, 2H), 1.33(d, J=1.4Hz, 9H).

[0553] Compound BB1-5 (4 g, 6.34 mmol) was dissolved in DMF (50 mL), and allyl bromide (843.99 mg, 6.98 mmol) and potassium carbonate (1.75 g, 12.68 mmol) were added. The system was stirred at room temperature (25 °C) for 16 h. Water (50 mL) was added to the system, and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 0–50%) to give compound BB1-6. MS m / z (ESI): 671.5 [M+H] + .

[0554] Compound BB1-6 (2.8 g, 4.17 mmol) was dissolved in DCM (10 mL) and TFA (10 mL). The system was stirred at room temperature (25 °C) for 16 h. The system was concentrated to obtain a crude product, which was purified by C18 reversed-phase medium-pressure column chromatography (water / acetonitrile = 0–50%) to give the intermediate Fmoc-Asp-Pal-Sar-OAll. MS m / z (ESI): 615.3 [M+H] + . 1H-NMR (400MHz, DMSO-d6) δ12.35 (s, 1H), 8.44-8.33 (m, 2H), 8.21 (m, 1H), 7.90 (d, J=7. 8Hz, 2H), 7.72 (d, J=7.8Hz, 2H), 7.65-7.56 (m, 2H), 7.42 (t, J=7.4Hz, 2H), 7.33 (t, J=7 .4Hz, 2H), 7.21 (dd, J=7.4, 4.8Hz, 1H), 5.97-5.85 (m, 1H), 5.36-5.20 (m, 2H), 4.98-4. 74(m, 1H), 4.68-4.57(m, 2H), 4.36-4.00(m, 6H), 3.11-2.78(m, 5H), 2.46-2.40(m, 2H).

[0555] Intermediate AA4

[0556] Compound AA4-1 (4.5 g, 14.9 mmol) and methyl 4-bromobutyrate (1.34 g, 7.45 mmol) were dissolved in acetonitrile (30 mL). The system was stirred at room temperature (25 °C) for 2.5 h, then heated to 80 °C and stirred for 16 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 0–5%) to give compound AA4-2. MS m / z (ESI): 371.4 [M+H] + .

[0557] Compound AA4-2 (2.7 g, 7.3 mmol) was dissolved in dichloromethane (10 mL), and 4 M HCl dioxane solution (5.5 mL, 21.9 mmol) was added. The system was stirred at room temperature (25 °C) for 2.5 h. The system was concentrated to give compound AA4-3, which was used directly in the next reaction without further purification. MS m / z (ESI): 215.2 [M+H] + .

[0558] The crude compound AA4-3 (7.3 mmol) was dissolved in a 1 / 1, v / v mixed solvent (20 mL) of acetonitrile / water. 9-fluorenylmethyl N-succinimide carbonate (3.2 g, 9.5 mmol) and sodium bicarbonate (3.1 g, 36.5 mmol) were added to the solution. The system was stirred at room temperature (25 °C) for 4 h. The pH was adjusted to 5 with 10% citric acid aqueous solution. The mixture was extracted twice with ethyl acetate (150 mL × 2). The combined organic phases were washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.05% trifluoroacetic acid) = 0–65%) to obtain intermediate AA4. MS m / z (ESI): 437.4 [M] + . 1HNMR (400MHz, DMSO) δ12.55 (s, 1H), 7.90 (d, J=7.5Hz, 2H), 7.73 (d, J=7.4Hz, 2H), 7.62 (d, J=8.0Hz, 1H), 7.42 (td, J=7.5, 1.1Hz, 2H), 7.33 (td, J=7.5, 1.2Hz, 2H), 4.34-4.17 ( m, 3H), 3.97-3.85 (m, 1H), 3.29 (t, J=7.1Hz, 2H), 3.15 (t, J=7.0Hz, 2H), 2.18 (t, J=8.0H z, 2H), 1.88 (p, J=7.7Hz, 2H), 1.77-1.54 (m, 2H), 1.51-1.36 (m, 2H), 1.33-1.20 (m, 2H).

[0559] Intermediate AA10

[0560] Compounds AA10-1 (6 g, 19.48 mmol) and AA10-2 (3.75 g, 23.38 mmol) were dissolved in MeOH (60 mL), and TEA (6.11 g, 60.40 mmol, 8.42 mL) was added. The system was heated to 70 °C and stirred under reflux for 12 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 0–33%) to give compound AA10-3. MS m / z (ESI): 377.2 [M+Na] + . 1 H-NMR (400MHz, DMSO-d6) δ7.67 (d, J=8.0Hz, 1H), 7.60-7.56 (m, 2H), 4.95 (brs , 1H), 4.45 (s, 2H), 3.71 (t, J=5.6Hz, 2H), 3.42 (q, J=6.0Hz, 2H), 1.31 (s, 9H).

[0561] Starting from AA10-3, synthesize intermediate AA10 by referring to the operation steps of intermediate AA1.

[0562] AA10: MS m / z(ESI): 486.3[M+H-Boc] + . 1 H-NMR (400MHz, DMSO-d6) δ12.82 (brs, 1H), 7.86 (d, J=7.6Hz, 2H), 7.75 (d, J=8.4Hz, 1H), 7.65-7.54 (m, 3H), 7.46-7.35 (m, 4H), 7. 31-7.24(m, 2H), 6.91(t, J=5.2Hz, 1H), 4.44-4.33(m, 2H), 4.25-4.12(m, 4H), 3.54-3.45(m, 2H), 3.21-2.93(m, 4H), 1.27(s, 9H).

[0563] Intermediate B1

[0564] 4-(4-iodophenyl)butyric acid (5.3 g, 18.2 mmol), Fmoc-S-lysine tert-butyl hydrochloride (7 g, 15.2 mmol), and DIEA (7 g, 45.4 mmol) were dissolved in DMF (100 mL). HATU (9.8 g, 18.2 mmol) was added at 0 °C, and the mixture was stirred at 0 °C for 1 h. Water (300 mL) was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (petroleum ether / ethyl acetate (v / v) = 10%) to give compound B1-1. MS m / z (ESI): 697.4 [M+H] + .

[0565] Compound B1-1 (10 g, 14.3 mmol) and DBU (2.4 g, 15.7 mmol) were dissolved in DCM (100 mL); the system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to give intermediate B1-2. MS m / z (ESI): 475.4 [M+H] + .

[0566] Compound B1-2 (900 mg, 1.90 mmol) was dissolved in tetrahydrofuran (12 mL). Triethylamine (479.95 mg, 4.74 mmol, 661 μL) and compound B1-3 (208.85 mg, 2.09 mmol) were added at room temperature (25 °C). The system was stirred at room temperature (25 °C) for 2 h. Saturated ammonium chloride (15 mL) was added to the system, and the mixture was extracted with ethyl acetate (12 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 5–35%) to obtain compound B1. MS m / z (ESI): 589.2 [M+H] + .

[0567] Starting from the corresponding starting materials, intermediate B12 was synthesized by referring to the operation steps of intermediate B1.

[0568] B12: MS m / z(ESI): 575.4[M+H] + .

[0569] Intermediate B2

[0570] Compound B2-1 (5 g, 20.6 mmol) was dissolved in THF (50 mL), and NBS (13.3 g, 40.11 mmol) and TPP (10.52 g, 40.11 mmol) were added. The system was stirred at room temperature (25 °C) for 2 h under nitrogen atmosphere. The system was then poured into ice water and stirred until homogeneous, followed by extraction three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 20%) to give compound B2-2. MS m / z (ESI): 311.5 [M+H] + .

[0571] Compound B2-2 (4 g, 12.81 mmol) was dissolved in acetonitrile (50 mL), and trimethylamine (20 mL) was added. Under nitrogen atmosphere, the system was heated to 50 °C and stirred for 16 h. The system was concentrated to obtain compound B2-3, which was used directly in the next reaction without further purification. MS m / z (ESI): 291.4 [M] + .

[0572] Compound B2-3 (4.0 g) was dissolved in DCM (50 mL), and TFA (20 mL) was added. The system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to obtain a crude product, which was purified by C18 reversed-phase medium-pressure column chromatography (water = 100%) to obtain intermediate B2. MS m / z (ESI): 191.2 [M]+. 1H NMR (400MHz, DMSO-d6) δ8.11 (s, 2H), 3.81 (m, 2H), 3.50-3.78 (m, 8H), 3.11 (s, 9H), 2.94 (s, 2H).

[0573] Intermediate B3

[0574] Compound B3-1 (30 g, 65.1 mmol) and sodium bicarbonate (21.9 g, 260.3 mmol) were dissolved in dichloromethane (100 mL) and water (200 mL). At room temperature (25 °C), a solution of Alloc-OSu (14.3 g, 71.6 mmol) in dichloromethane (100 mL) was slowly added dropwise. The system was stirred at room temperature (25 °C) for 1 h. After washing with saturated sodium chloride solution, the mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 5%) to obtain compound B3-2. MS m / z (ESI): 509.42 [M+H] + .

[0575] Compound B3-2 (33 g, 64.9 mmol) and DBU (12.8 g, 84.4 mmol) were dissolved in dichloromethane (400 mL); the system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to give compound B3-3. MS m / z (ESI): 287.29 [M+H] + .

[0576] Compound B3-3 (20 g, 69.8 mmol) and 37% wt formaldehyde aqueous solution (20 mL) were dissolved in methanol (200 mL). At 0 °C, sodium triacetylborohydride (44.4 g, 209.5 mmol) was added, and the mixture was stirred at 0 °C for 1 h. Sodium hydroxide aqueous solution (0.5 N, 300 mL) was added, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to give compound B3-4. MS m / z (ESI): 315.29 [M+H] + .

[0577] Compound B3-4 (13.8 g, 43.9 mmol), tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethyl)carbamate (41.1 g, 131.7 mmol), and potassium carbonate (18.2 g, 131.7 mmol) were dissolved in acetonitrile (200 mL); the system was heated to 80 °C and stirred for 6 h. The system was filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to obtain compound B3-5. MS m / z (ESI): 546.46 [M] + .

[0578] Compound B3-5 (18 g, 28.7 mmol) was dissolved in DCM (100 mL), and TFA (200 mL) was added at room temperature (25 °C). The system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to give compound B3-6, which was used directly in the next reaction without further purification. MS m / z (ESI): 390.31 [M] + .

[0579] Compound B3-6 (13.5 g, 28.7 mmol) and sodium bicarbonate (12.1 g, 143.5 mmol) were dissolved in acetonitrile (100 mL) and water (100 mL). Fmoc-OSu (11.6 g, 34.4 mmol) was added at 0 °C. The system was stirred at room temperature (25 °C) for 2 h. Hydrochloric acid (6 M) was added to adjust the pH to 4. The system was purified by C18 reversed-phase medium-pressure column chromatography (water (0.3% HCl) / acetonitrile = 67%) to give compound B3. MS m / z (ESI): 612.37 [M] + .

[0580] Intermediate B5

[0581] Oxaloyl chloride (3.93 g, 30.94 mmol) was dissolved in dichloromethane (38 mL). At -70 °C, a dichloromethane solution of DMSO (3.78 g, 48.43 mmol) was added dropwise (2 mL). The system was stirred at -70 °C for 1 h. At -70 °C, a dichloromethane solution of compound B5-1 (4 g, 13.45 mmol) was added (25 mL). The system was stirred at -70 °C for 1 h. At -70 °C, triethylamine (8.17 g, 80.71 mmol) was slowly added. The system was stirred at -70 °C for 10 min, then heated to room temperature (25 °C) and stirred for 40 min. The mixture was quenched with water (30 mL) and extracted three times with dichloromethane (25 mL). The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 25%) to give compound B5-2. MS m / z (ESI): 296.2 [M+H] + .

[0582] Compound B5-2 (1.8 g, 6.09 mmol) and compound B5-3 (1.42 g, 6.09 mmol) were dissolved in tetrahydrofuran (36 mL); the system was stirred at 15 °C for 0.5 h. Sodium borohydride acetate (1.29 g, 6.09 mmol) was added; the system was stirred at 15 °C for 0.5 h. The system was filtered, and the filtrate was concentrated to obtain a crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 50%) to give compound B5-4. MS m / z (ESI): 513.5 [M+H] + .

[0583] Compound B5-4 (600.00 mg, 995.79 μmol) was dissolved in acetonitrile (8 mL), and potassium carbonate (323.52 mg, 2.34 mmol) and methyl iodide (830.64 mg, 5.85 mmol) were added. The system was stirred at 30 °C for 1 h. The system was filtered, and the filtrate was concentrated to obtain compound B5-5, which was used directly in the next reaction without further purification. MS m / z (ESI): 541.5 [M] + .

[0584] Compound B5-5 (600.00 mg, 995.79 μmol) was dissolved in dioxane hydrochloride solution (4 M, 12 mL); the system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to obtain compound B5, which was used directly in the next reaction without further purification. MS m / z (ESI): 485.4 [M] + .

[0585] Starting from the corresponding starting materials, intermediate B4 was synthesized by referring to the operation steps of intermediate B5.

[0586] B4: MS m / z (ESI): 456.5 [M] + .

[0587] Intermediate B6

[0588] Compound B6-1 (5 g, 15.95 mmol) was dissolved in DCM (60 mL), and TEA (4.84 g, 47.86 mmol) was added. Methylsulfonyl chloride (3.69 g, 19.15 mmol) was then added dropwise at 0 °C. The system was stirred at 18 °C for 1.5 h. After washing with saturated sodium chloride solution, the mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound B6-2, which was used directly in the next reaction without further purification. MS m / z (ESI): 392.3 [M+H] + .

[0589] Compound B6-3 (3 g, 14.83 mmol) and aq. HCHO (6.83 g, 74.15 mmol, 37% purity) were dissolved in MeOH (40 mL), and AcOH (890.57 mg, 14.83 mmol) was added. The system was stirred at room temperature (25 °C) for 20 min, and NaCNBH3 (2.80 g, 44.49 mmol) was added. The system was stirred at room temperature (25 °C) for 16 h. Water (25 mL) was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (tetrahydrofuran / petroleum ether (v / v) = 30%-66%) to obtain compound B6-4. MS m / z (ESI): 231.2 [M+H] + .

[0590] Compound B6-4 (2 g, 8.68 mmol) and compound B6-2 (3.40 g, 8.68 mmol) were dissolved in ACN (40 mL), and K2CO3 (3.60 g, 26.05 mmol) and KI (288.26 mg, 1.74 mmol) were added. The system was heated to 90 °C and stirred for 5 h. The system was filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% TFA) = 0–33%) to obtain compound B6-5. MS m / z (ESI): 526.5 [M] + .

[0591] Compound B6-5 (1.54 g, 2.93 mmol) was dissolved in dioxane hydrochloride (15 mL); the system was stirred at room temperature (25 °C) for 1 h. The system was concentrated to obtain compound B6, which was used directly in the next reaction without further purification. 1 H-NMR (400MHz, DMSO-d6) 63.90-3.81 (m, 2H), 3.70-3.61 (m, 8H), 3.53-3.52 (m, 2H), 3.34-3.27 (m, 2H), 3.07 ( s, 6H), 2.81 (t, J=7.6Hz, 2H), 2.49 (s, 3H), 2.30 (s, 4H), 1.74-1.57 (m, 4H), 1.40-1.25 (m, 2H), 0.96 (s, 6H).MS m / z(ESI):426.4[M] + .

[0592] Starting from the corresponding starting materials, intermediate B14 is synthesized following the same procedure as intermediate B6.

[0593] B14: MS m / z (ESI): 428.4 [M] + .

[0594] Intermediate B7

[0595] Compound B7-1 (5 g, 14.3 mmol) was dissolved in MeOH (50 mL) and DCM (10 mL), and thionyl chloride (1.56 mL, 21.5 mmol) was added at 0 °C. The system was stirred at room temperature (25 °C) for 16 h. The system was concentrated to give compound B7-2, which was used directly in the next reaction without further purification. 1 H NMR (400MHz, DMSO) δ3.57 (s, 3H), 3.51 (t, J = 6.8Hz, 2H), 2.28 (t, J = 7.2Hz, 2 H), 1.83-1.73(m, 2H), 1.54-1.48(m, 2H), 1.40-1.34(m, 2H), 1.24(s, 24H).

[0596] Dibenzyl phosphite (4 g, 15.1 mmol) was dissolved in DMF (50 mL), and sodium hydride (0.55 g, 15.1 mmol) was added at 0 °C. The system was stirred at 0 °C for 0.5 h. B7-1 (5 g, 13.8 mmol) was added at 0 °C. The system was stirred at room temperature (25 °C) for 1 h. Water (200 mL) was added to the system, and the mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 25%) to give compound B7-3. MS m / z (ESI): 545.4 [M+H] + .

[0597] Compound B7-3 (6.8 g, 12.5 mmol) was dissolved in MeOH (60 mL) and H₂O (15 mL). Sodium hydroxide (1.5 g, 37.5 mmol) was added at 0 °C. The system was stirred at room temperature (25 °C) for 3 h. Water (100 mL) was added, and the pH was adjusted to neutral with 1 mol / L hydrochloric acid. The mixture was extracted three times with ethyl acetate. The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to obtain compound B7. MS m / z (ESI): 531.4 [M+H] + .

[0598] Intermediate B8

[0599] Compound B8-1 (1.7 g, 9.1 mmol), compound B8-2 (2.4 g, 9.1 mmol), and potassium carbonate (5 g, 36.4 mmol) were dissolved in DMF (50 mL); the system was stirred at 60 °C for 2 h. Water (100 mL) was added to the system, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (ethyl acetate / petroleum ether (v / v) = 50%) to give compound B8-3. MS m / z (ESI): 373.3 [M+H] + .

[0600] Compound B8-3 (2.1 g, 5.5 mmol) was dissolved in DCM (20 mL), and a 4 M HCl solution of dioxane (20 mL) was added at room temperature (25 °C). The system was stirred at room temperature (25 °C) for 2 h. The system was concentrated to obtain compound B8-4, which was used directly in the next reaction without further purification. MS m / z (ESI): 217.2 [M+H] + .

[0601] Compound B8-4 (1.2 g, 5.5 mmol) and sodium bicarbonate (2.3 g, 27.5 mmol) were dissolved in acetonitrile (15 mL) and water (15 mL). 9-fluorenylmethyl-N-succinimide carbonate (2.2 g, 6.6 mmol) was slowly added dropwise at 0 °C. The system was stirred at room temperature (25 °C) for 2 h. 1 M hydrochloric acid aqueous solution (50 mL) was added, and the system was purified by C18 reversed-phase medium-pressure column chromatography ((0.5% TFA) water / acetonitrile = 67%) to give compound B8. MS m / z (ESI): 439.3 [M+H] + .

[0602] Starting from the corresponding starting materials, intermediate B9 was synthesized by referring to the operation steps of intermediate B8.

[0603] B9: MS m / z (ESI): 411.3 [M+H] + .

[0604] Intermediate B10

[0605] Compound B10-1 (1 g, 3.5 mmol) was dissolved in formic acid (10 mL), and hydrogen peroxide (2.5 mL, 22.7 mmol, 30% wt) was added at 0 °C. The system was stirred at room temperature (25 °C) for 16 h. The system was freeze-dried to obtain compound B10, which was used directly in the next reaction without further purification. MS m / z (ESI): 335.2 [MH] + .

[0606] Intermediate B11

[0607] Compound B11-1 (5 g, 16.82 mmol) was dissolved in ACN (50 mL), and trimethylamine (2.98 g, 50.47 mmol) was added. Under a nitrogen atmosphere, the system was heated to 50 °C and stirred for 16 h. The system was concentrated to obtain compound B11-2, which was used directly in the next reaction without further purification. MS m / z (ESI): 276.2 [M] + .

[0608] Compound B11-2 (3.5 g, 12.66 mmol) was dissolved in dichloromethane (50 mL), and TFA (25 mL) was added. The mixture was stirred at room temperature (25 °C) for 2 h under a nitrogen atmosphere. The system was concentrated to obtain a crude product, which was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 10%) to give compound B11. MS m / z (ESI): 220.2 [M] + .1 HNMR (400MHz, DMSO-d6) δ4.09-4.01 (m, 2H), 3.61-3.53 (m, 8H), 3.11 (s, 9H), 2.45 (t, J=6.0Hz, 2H).

[0609] Intermediate B13

[0610] Compound B13-1 (5 g, 30 mmol) and concentrated sulfuric acid (1.5 g, 7.5 mmol) were dissolved in methanol (50 mL), and the system was stirred at 40 °C for 16 h. Water (100 mL) was added to the system, and the mixture was extracted three times with DCM. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound B13-2, which was used directly in the next reaction without further purification. MS m / z (ESI): 335.2 [MH] + .

[0611] 18-(tert-butoxy)-18-oxooctadecanoic acid (2 g, 5.5 mmol) and CDI (0.94 g, 5.8 mmol) were dissolved in DCM (10 mL), and the system was stirred at room temperature (25 °C) for 2 h. Compound B13-2 (1 g, 5.5 mmol) and DBU (1.7 g, 11 mmol) were added, and the system was stirred at room temperature (25 °C) for 16 h. The reaction solution was poured into citric acid buffer solution (pH = 4.5), filtered, and the filter cake was washed with water and dried to obtain compound B13-3. ESI-MS m / z = 478.4 [M+H- t Bu] + .

[0612] Compound B13-3 (0.84 g, 1.6 mmol) and sodium hydroxide (0.19 g, 4.8 mmol) were added to tetrahydrofuran (10 mL), methanol (2 mL), and water (2 mL); the system was stirred at room temperature (25 °C) for 4 h. Water (100 mL) was added to the system, and the pH was adjusted to 7 with 1 M hydrochloric acid. The mixture was extracted three times with ethyl acetate. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by medium-pressure column chromatography (methanol / dichloromethane (v / v) = 7%) to obtain compound B13. MS m / z (ESI): 464.4 [M+H- t Bu] + .

[0613] Example 2: Synthesis of polypeptide compounds

[0614] In the following polypeptide synthesis, Rink amide-MBHA resin is... 2-CTC Resin is

[0615] Synthetic route one:

[0616] Synthesis of Compound 1:

[0617] Chemical solid-phase synthesis:

[0618] 1.1 Composite Table:

[0619] 1.2 Operating Steps:

[0620] 1.2.1 Weigh 1.5g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After the system is filtered, wash the resin twice with 10V DMF.

[0621] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0622] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0623] 1.2.4 Weigh 3.0 eq Fmoc-Lys(Alloc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0624] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0625] 1.2.6 Add 10V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0626] 1.2.7 Weigh 3.0eq t BuO-Ste-Glu(AEEA-AEEA-OH)-O t Bu, 3.0 eq Oxyma, 3.0 eq DIC, and 10 V DMF were added to the reactor, and N2 was blown in for 1 hour.

[0627] 1.2.8 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0628] 1.2.9 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 3-15 in sequence.

[0629] 1.2.10 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0630] 1.2.11 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0631] 1.2.12 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0632] 1.2.13 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0633] 1.3 Cutting and pyrolysis:

[0634] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), and lyse at room temperature for 2.5 h. Filter. At 0℃, add 10V MTBE to the filtrate, causing solid precipitation. Centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 1-2.

[0635] 1.4 Disulfide bond cyclization:

[0636] Weigh crude peptides 1-2 and add them to a reaction flask. Dissolve them in 50% methanol / water solution to prepare a 40 mg / mL solution. Add 1.5 eq I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 10-15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. Purify the system by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 1.

[0637] Salt conversion conditions:

[0638] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0639] MS m / z(ESI): 1391.7[(M+2H) / 2] + 928.4[(M+3H) / 3] + .

[0640] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0641] Synthetic route two:

[0642] Synthesis of compound 2:

[0643] Chemical solid-phase synthesis:

[0644] 1.1 Composite Table:

[0645] 1.2 Operating Steps:

[0646] 1.2.1 Weigh 1.5g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After the system is filtered, wash the resin twice with 10V DMF.

[0647] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0648] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0649] 1.2.4 Weigh 3.0 eq Fmoc-Sar-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0650] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0651] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-14 in sequence.

[0652] 1.2.7 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0653] 1.3 Cutting and pyrolysis:

[0654] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), and lyse at room temperature for 2.5 h. Filter. At 0℃, add 10V MTBE to the filtrate, causing solid precipitation. Centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 2-2.

[0655] 1.4 Disulfide bond cyclization:

[0656] Weigh crude peptide 2-2 and add it to a reaction flask. Dissolve it in 50% methanol / water solution to prepare a 40 mg / mL solution. Add 1.5 eq I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 10-15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. Purify the system by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 2.

[0657] Salt conversion conditions:

[0658] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0659] MS m / z(ESI): 1306.7[(M+2H) / 2] + 871.7[(M+3H) / 3] + .

[0660] Synthetic route three:

[0661] Synthesis of compound 3:

[0662] Chemical solid-phase synthesis:

[0663] 1.1 Composite Table:

[0664] 1.2 Operating Steps:

[0665] 1.2.1 Weigh 1.46g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0666] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0667] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0668] 1.2.4 Weigh 2.0 eq Fmoc-Sar-OH, 4.0 eq DIEA, and 10 V DMF and add them to the reactor. Then add 2.0 eq HATU and purge with N2 for 1 h.

[0669] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0670] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-17 in sequence.

[0671] 1.2.7 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0672] 1.3 Cutting and pyrolysis:

[0673] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / ml)), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide 3-2.

[0674] 1.4 Disulfide bond cyclization:

[0675] Weigh crude peptide 3-2 and add it to a reaction flask. Dissolve it in a 10%-20% acetonitrile / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. Add 0.2 mmol / mL I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 10-15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. The system is purified by high-performance liquid chromatography and converted to salt to obtain compound 3.

[0676] Salt conversion conditions:

[0677] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0678] MS m / z(ESI): 1298.8[(M+2H) / 2] + 866.1[(M+3H) / 3] + .

[0679] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0680] Synthetic route four:

[0681] Synthesis of compound 5:

[0682] 1.2 Operating Steps:

[0683] 1.2.1 Weigh 1.45g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0684] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0685] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0686] 1.2.4 Weigh 2.0 eq Fmoc-Asp-Pal-Sar-OAll, 4.0 eq DIEA, and 10V DMF and add them to the reactor, then add 2.0 eq HATU and purge with N2 for 1 hour.

[0687] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 6 times with 10V DMF.

[0688] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-11 in sequence.

[0689] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0690] 1.2.8 Add 3 mL Ac2O, 6 mL DIEA, 10 V DMF to the reactor and purge with N2 for 1 h; after the reaction is complete, the ninhydrin / tetrachlorobenzoquinone test is negative; after filtration, wash the resin 6 times with 10 V DMF.

[0691] 1.2.9 Add DCM swelling resin to the reactor for 30 min, then add phenylsilane (20 eq), 1,3-dimethylbarbituric acid (20 eq), tetra-triphenylphosphine palladium (0.5 eq), and purge with N2 for 6 h; after the reaction is completed, the ninhydrin / tetrachlorobenzoquinone test is negative; after the system is filtered, the resin is washed 6 times with 10V DMF.

[0692] 1.2.10 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 13-16 in sequence.

[0693] 1.2.11 Wash the resin twice with MeOH and twice with MTBE, then dry the resin until it becomes fine sand.

[0694] 1.3 Cutting and pyrolysis:

[0695] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / ml)), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide 5-4.

[0696] 1.4 Disulfide bond cyclization:

[0697] Crude peptide 5-4 was weighed and added to a reaction flask, dissolved in a 10%-20% acetonitrile / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. 0.2 mmol / mL I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color of the reaction solution disappeared. The system was purified by high-performance liquid chromatography and converted to salt to obtain compound 5.

[0698] Salt conversion conditions:

[0699] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0700] MS m / z(ESI): 1312.6[(M+2H) / 2] + 875.4[(M+3H) / 3] + .

[0701] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0702] Synthetic route five:

[0703] Synthesis of compound 9:

[0704] Chemical solid-phase synthesis:

[0705] 1.1 Composite Table:

[0706] 1.2 Operating Steps:

[0707] 1.2.1 Weigh 1.5g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After the system is filtered, wash the resin twice with 10V DMF.

[0708] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0709] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0710] 1.2.4 Weigh 3.0 eq Fmoc-Lys(Alloc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0711] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0712] 1.2.6 Add 10V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0713] 1.2.7 Weigh 3.0eq t BuO-Ste-Glu(AEEA-AEEA-OH)-O t Bu, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF were added to the reactor, and N2 was blown in for 1 hour.

[0714] 1.2.8 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0715] 1.2.9 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 3-15 in sequence.

[0716] 1.2.10 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0717] 1.2.11 Weigh 3.0 eq intermediate B1, 3.0 eq HATU, 6.0 eq IDEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0718] 1.2.12 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0719] 1.2.13 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0720] 1.3 Cutting and pyrolysis:

[0721] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 9-2.

[0722] 1.4 Disulfide bond cyclization:

[0723] Weigh crude peptide 9-2 and add it to a reaction flask. Dissolve it in 50% methanol / water solution to prepare a 40 mg / mL solution. Add 1.5 eq I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 10-15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. Purify the system by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 9.

[0724] Salt conversion conditions:

[0725] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0726] MS m / z(ESI): 1648.84[(M+H) / 2] + 1099.66[(M+2H) / 3] + .

[0727] Synthetic route six:

[0728] Synthesis of compound 10:

[0729] Chemical solid-phase synthesis:

[0730] 1.1 Composite Table:

[0731] 1.2 Operating Steps:

[0732] 1.2.1 Weigh 1.5g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After the system is filtered, wash the resin twice with 10V DMF.

[0733] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0734] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0735] 1.2.4 Weigh 3.0 eq Fmoc-Lys(Dde)-OH, 4.0 eq DIEA, and 10 V DMF and add them to the reactor. Then add 2.0 eq HATU to the reactor and purge with N2 for 1 h.

[0736] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0737] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-14 in sequence.

[0738] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0739] 1.2.8 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0740] 1.2.9 Add 3 mL Ac2O, 6 mL DIEA, 10 V DMF to the reactor and purge with N2 for 1 h; after the reaction is complete, the ninhydrin / tetrachlorobenzoquinone test is negative; after filtration, wash the resin 6 times with 10 V DMF.

[0741] 1.2.10 Add 10V 5% hydrazine hydrate / DMF to the reactor and purge with N2 for 15 min; repeat this operation three times.

[0742] 1.2.11 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0743] 1.2.12 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 16-19 in sequence.

[0744] 1.2.13 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0745] 1.2.14 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0746] 1.3 Cutting and pyrolysis:

[0747] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / mL)), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide 10⁻³.

[0748] 1.4 Disulfide bond cyclization:

[0749] Weigh 10⁻³ of crude peptide and add it to a reaction flask. Dissolve it in a 10%-20% acetonitrile / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. Add 0.2 mmol / mL I₂ / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. The system is purified by high-performance liquid chromatography and converted to salt to obtain compound 10.

[0750] Salt conversion conditions:

[0751] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0752] MS m / z (ESI): 918.1 [(M+3H) / 3] + .

[0753] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0754] Synthetic route seven:

[0755] Synthesis of compound 12:

[0756] Chemical solid-phase synthesis:

[0757] 1.1 Composite Table:

[0758] 1.2 Operating Steps:

[0759] 1.2.1 Weigh 1.45g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0760] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0761] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0762] 1.2.4 Weigh 3.0 eq Fmoc-Asp-Pal-Sar-OAll, 4.0 eq DIEA, and 10V DMF and add them to the reactor. Then add 2.0 eq HATU and N2 to the reactor and purge for 1 hour.

[0763] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0764] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-15 in sequence.

[0765] 1.2.7 Add DCM swelling resin to the reactor for 30 min, then add phenylsilane (20 eq), 1,3-dimethylbarbituric acid (20 eq), tetra-triphenylphosphine palladium (0.5 eq), and purge with N2 for 6 h; after the reaction is completed, the ninhydrin / tetrachlorobenzoquinone test is negative; after the system is filtered, the resin is washed 6 times with 10V DMF.

[0766] 1.2.8 Add 10V DMF, 2.0eq intermediate B2, and 4.0eq DIEA to the reactor, then add 2.0eq HATU and N2 and purge for 1h.

[0767] 1.2.9 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0768] 1.2.10 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0769] 1.3 Cutting and pyrolysis:

[0770] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / ml)), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide 12-3.

[0771] 1.4 Disulfide bond cyclization:

[0772] Crude peptide 12-3 was weighed and added to a reaction flask, dissolved in a 10%-20% acetonitrile / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. 0.2 mmol / mL I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color of the reaction solution disappeared. The system was purified by high-performance liquid chromatography and converted to salt to obtain compound 12.

[0773] Salt conversion conditions:

[0774] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0775] MS m / z (ESI): 923.8 [(M+2H) / 3] + 693.2[(M+3H) / 4] + ;

[0776] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0777] Synthetic route eight:

[0778] Synthesis of compound 24:

[0779] Chemical solid-phase synthesis:

[0780] 1.1 Composite Table:

[0781] 1.2 Operating Steps:

[0782] 1.2.1 Weigh 2.5g of Rink amide-MBHA resin (degree of substitution 0.361mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0783] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0784] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0785] 1.2.4 Weigh 3.0 eq intermediate B3, 3.0 eq HATU, 6.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0786] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0787] 1.2.6 Repeat steps 1.2.2-1.2.5 of the synthesis table to condense amino acid 2.

[0788] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0789] 1.2.8 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0790] 1.2.9 Weigh 3.0eq t BuO-Ste-Glu(O t Bu)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF were added to the reactor, and N2 was blown in for 0.5 h.

[0791] 1.2.10 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0792] 1.2.11 Add 10V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0793] 1.2.7 Weigh 3.0 eq Fmoc-Sar-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0794] 1.2.8 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0795] 1.2.9 Repeat steps 1.2.7-1.2.10 according to the synthesis table to condense amino acids 5-16 in sequence.

[0796] 1.2.10 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0797] 1.2.11 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0798] 1.2.12 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0799] 1.2.13 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0800] 1.3 Cutting and pyrolysis:

[0801] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 24-2.

[0802] 1.4 Disulfide bond cyclization:

[0803] Crude peptide 24-2 was weighed and added to a reaction flask, dissolved in 50% methanol / water solution to prepare a 40 mg / mL solution. 1.5 eq I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 10-15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 24.

[0804] Salt conversion conditions:

[0805] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0806] MS m / z(ESI): 1403.18[(M+H) / 2] + 935.97[(M+2H) / 3] + .

[0807] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0808] Synthetic route nine:

[0809] Synthesis of Compound 50:

[0810] Chemical solid-phase synthesis:

[0811] 1.1 Composite Table:

[0812] 1.2 Operating Steps:

[0813] 1.2.1 Weigh 1g of Rink amide-MBHA resin (degree of substitution 0.318mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0814] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0815] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0816] 1.2.4 Weigh 2.0 eq intermediate AA13, 4.0 eq DIEA, and 10 V DMF and add them to the reactor, then add 2.0 eq HATU and N2 and purge for 1 h.

[0817] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 6 times with 10V DMF.

[0818] 1.2.6 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0819] 1.2.7 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0820] 1.2.8 Weigh 2.0 eq of intermediate Fmoc-Glu(O) t Bu)-OH, 2.0 eq Oxyma, 2.0 eq DIC, 10 V DMF, activated for 10 min, then added to the reactor, and N2 was blown in for 1 h.

[0821] 1.2.9 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 6 times with 10V DMF.

[0822] 1.2.10 Repeat steps 1.2.6-1.2.9 according to the synthesis table to condense amino acids 3-11 in sequence.

[0823] 1.2.11 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0824] 1.2.12 Add 10V DMF to the reactor, then add 4.0eq DIEA and 2.0eq AC2O, and react for 1h under nitrogen protection; after the reaction is completed, the ninhydrin / tetrachlorobenzoquinone test is negative; after the system is filtered, the resin is washed 6 times with 10V DMF.

[0825] 1.2.13 Add DCM swelling resin to the reactor for 30 min, then add phenylsilane (20 eq), 1,3-dimethylbarbituric acid (20 eq), tetratetraphenylphosphine palladium (0.5 eq), and purge with N2 for 5 h; after the reaction is completed, the ninhydrin / tetrachlorobenzoquinone test is negative; after the system is filtered, the resin is washed 6 times with 10V DMF.

[0826] 1.2.14 Weigh 2.0 eq intermediate B6, 4.0 eq DIEA, and 10 V DMF and add them to the reactor, then add 2.0 eq HATU and N2 and purge for 1 h.

[0827] 1.2.15 After the reaction is complete, wash the resin 6 times with 10V DMF.

[0828] 1.2.16 Add 10V 5% hydrazine hydrate / DMF to the reactor and purge with N2 for 15 min; repeat this operation three times.

[0829] 1.2.17 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0830] 1.2.18 Weigh 2.0 eq intermediate Fmoc-AEEA-OH, 2.0 eq Oxyma, 2.0 eq DIC, 10 V DMF, activate for 10 min and add to the reactor, then purge with N2 for 1 h.

[0831] 1.2.19 The reaction ended, and the ninhydrin / tetrachlorobenzoquinone test was negative. After filtration, the resin was washed 6 times with 10V DMF.

[0832] 1.2.20 Repeat steps 1.2.6-1.2.9 according to the synthesis table to condense amino acids 15-16 in sequence.

[0833] 1.2.15 Wash the resin twice with MeOH and twice with MTBE, then dry the resin until it becomes fine sand.

[0834] 1.3 Cutting and pyrolysis:

[0835] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / ml)), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide 50-5.

[0836] 1.4 Disulfide bond cyclization:

[0837] Weigh crude peptide 50-5 and add it to a reaction flask. Dissolve it in a 10%-20% acetonitrile / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. Add 0.2 mmol / mL I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. The system is purified by high-performance liquid chromatography and converted to salt to obtain compound 50.

[0838] Salt conversion conditions:

[0839] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0840] MS m / z (ESI): 1382.4 [(M+H) / 2] + ; 921.8[(M+2H) / 3] + .

[0841] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0842] Synthetic route ten:

[0843] Synthesis of compound 57:

[0844] Chemical solid-phase synthesis:

[0845] 1.1 Composite Table:

[0846] 1.2 Operating Steps:

[0847] 1.2.1 Weigh 1.5g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After the system is filtered, wash the resin twice with 10V DMF.

[0848] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0849] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0850] 1.2.4 Weigh 3.0 eq Alloc-Lys(Fmoc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0851] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0852] 1.2.6 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0853] 1.2.7 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0854] 1.2.8 Weigh 3.0eq t BuO-Ste-Glu(AEEA-AEEA-OH)-O t Bu, 3.0 eq HATU, 6.0 eq DIEA, and 10 V DMF were added to the reactor, and N2 was blown in for 1 hour.

[0855] 1.2.9 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0856] 1.2.10 Add 10V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0857] 1.2.11 Weigh 3.0 eq Fmoc-Sar-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0858] 1.2.12 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0859] 1.2.13 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 4-16 in sequence.

[0860] 1.2.14 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0861] 1.2.15 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 40 min.

[0862] 1.2.16 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0863] 1.2.17 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0864] 1.3 Cutting and pyrolysis:

[0865] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), lyse at room temperature for 2.5 h, and filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates, centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 57-2.

[0866] 1.4 Disulfide bond cyclization:

[0867] Crude peptide 57-2 was weighed and added to a reaction flask, dissolved in 50% methanol / water solution to prepare a 40 mg / mL solution. 1.5 eq I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 10-15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 57.

[0868] Salt conversion conditions:

[0869] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0870] MS m / z(ESI): 1475.41[(M+2H) / 2] + 983.68[(M+3H) / 3] + .

[0871] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0872] Synthetic route eleven:

[0873] Synthesis of compound 76:

[0874] Chemical solid-phase synthesis:

[0875] 1.1 Composite Table:

[0876] 1.2 Operating Steps:

[0877] 1.2.1 Weigh 1g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0878] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0879] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0880] 1.2.4 Weigh 3.0 eq Alloc-Lys(Fmoc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0881] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0882] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-5 in sequence.

[0883] 1.2.7 Add 10V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0884] 1.2.8 Weigh 3.0 eq Fmoc-Sar-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0885] 1.2.9 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0886] 1.2.10 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 7-18 in sequence.

[0887] 1.2.11 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0888] 1.2.12 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 40 min.

[0889] 1.2.13 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0890] 1.2.14 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0891] 1.3 Cutting and pyrolysis:

[0892] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), and lyse at room temperature for 2.5 h. Filter. At 0℃, add 10V MTBE to the filtrate, causing solid precipitation. Centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 76-2.

[0893] 1.4 Disulfide bond cyclization:

[0894] Crude peptide 76-2 was weighed and added to a reaction flask, dissolved in 50% methanol / water solution to prepare a 40 mg / mL solution. 1.5 eq I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 10-15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 76.

[0895] Salt conversion conditions:

[0896] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0897] MS m / z(ESI): 1415.06[(M+2H) / 2] + ; 943.61[(M+3H) / 3] + .

[0898] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0899] Synthetic route 12:

[0900] Synthesis of compound 83:

[0901] Chemical solid-phase synthesis:

[0902] 1.1 Composite Table:

[0903] 1.2 Operating Steps:

[0904] 1.2.1 Weigh 1g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0905] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0906] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0907] 1.2.4 Weigh 3.0 eq Fmoc-Lys(Alloc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0908] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0909] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-14 in sequence.

[0910] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0911] 1.2.8 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0912] 1.2.9 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0913] 1.2.10 Add 20V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF.

[0914] 1.2.11 Weigh 3.0 eq Fmoc-AEEA-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0915] 1.2.12 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0916] 1.2.13 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 17-19 in sequence.

[0917] 1.2.14 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0918] 1.3 Cutting and pyrolysis:

[0919] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole), and lyse at room temperature for 2.5 h. Filter. At 0℃, add 10V MTBE to the filtrate, causing solid precipitation. Centrifuge, wash the solid twice with MTBE, dissolve the centrifuged solid in acetonitrile, and purify by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain 83-2.

[0920] 1.4 Disulfide bond cyclization:

[0921] Crude peptide 83-2 was weighed and added to a reaction flask, dissolved in 50% methanol / water solution to prepare a 40 mg / mL solution. 1.5 eq I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 10-15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)), followed by high-performance liquid chromatography and salt transfer to obtain compound 83.

[0922] Salt conversion conditions:

[0923] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0924] MS m / z(ESI): 1407.84[(M+2H) / 2] + 939.69[(M+3H) / 3] + .

[0925] Starting from the corresponding starting materials, the following compounds were synthesized following the procedures described above:

[0926] Synthetic route thirteen:

[0927] Synthesis of compound 87:

[0928] Chemical solid-phase synthesis:

[0929] 1.1 Composite Table:

[0930] 1.2 Operating Steps:

[0931] 1.2.1 Weigh 3g of 2-CTC Resin (degree of substitution 1.13mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin three times with 10V DMF.

[0932] 1.2.2 Weigh 1.5 mmol Fmoc-Lys(Dde)-OH and add it to the reactor, then add 10 V DCM and 3 mmol DIEA, and purge with nitrogen for 3 h.

[0933] 1.2.3 After the reaction is complete, wash the resin three times with 10V DMF, add a 10V MEOH / DIEA / DMF (1 / 2 / 4) mixed solution, and purge with nitrogen for 30 minutes.

[0934] 1.2.4 After the reaction is complete, wash the resin 6 times with 10V DMF.

[0935] 1.2.5 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0936] 1.2.6 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0937] 1.2.7 Weigh 2.0 eq 18-(tert-butoxy)-18-oxooctadecanoic acid, 2.0 eq Oxyma, 2.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0938] 1.2.8 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 6 times with 10V DMF.

[0939] 1.2.9 Add 10V 5% hydrazine hydrate / DMF to the reactor, purge with N2 for 15 min, and repeat three times.

[0940] 1.2.10 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0941] 1.2.11 Weigh 2.0 eq Fmoc-AEEA-OH, 2.0 eq Oxyma, 2.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0942] 1.2.12 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0943] 1.2.13 Repeat steps 1.2.5-1.2.8 according to the synthesis table to condense amino acids 4-18 in sequence.

[0944] 1.2.14 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0945] 1.2.15 Weigh 6 mL Ac2O, 12 mL DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0946] 1.2.16 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0947] 1.2.17 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0948] 1.3 Cutting and pyrolysis:

[0949] Weigh the peptide resin into a centrifuge tube, add 10V 85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole sulfide + NH4I (150mg / mL) reagent, lyse at room temperature for 2.5h, and filter. Add 10V 0℃ MTBE to the filtrate, precipitate the solid, centrifuge, wash twice with MTBE, and dry the centrifuged solid with N2 to obtain 87-3.

[0950] 1.4 Disulfide bond cyclization:

[0951] Crude peptide 87-3 was weighed and added to a reaction flask, dissolved in a 10%-20% acetonitrile / water solution to prepare a 0.5-1 mg / mL solution. 0.2 mmol / mL I₂ / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by high-performance liquid chromatography and converted to salt to obtain compound 87.

[0952] Salt conversion conditions:

[0953] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0954] MS m / z(ESI): 1396.9[(M+2H) / 2] + ; 931.5[(M+3H) / 3] + .

[0955] Synthetic Route Fourteen:

[0956] Synthesis of peptide Pep01:

[0957] Chemical solid-phase synthesis:

[0958] Composite table:

[0959] 1.2 Solid-phase synthesis operation steps:

[0960] 1.2.1 Weigh 1.12g of Rink amide-MBHA resin (degree of substitution 0.268mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0961] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0962] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0963] 1.2.4 Weigh 2.0 eq Fmoc-Sar-OH, 4.0 eq DIEA, and 10 V DMF and add them to the reactor, then add 1.8 eq HATU and purge with N2 for 1 h.

[0964] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0965] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-13 in sequence.

[0966] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 10 min.

[0967] 1.2.8 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 40 min.

[0968] 1.2.9 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0969] 1.2.10 Wash the resin twice with MeOH and twice with DCM, then dry the resin until it becomes fine sand.

[0970] 1.3 Cutting and pyrolysis:

[0971] Weigh the peptide resin into a centrifuge tube, add 10V cleavage buffer (85% TFA / 2.5% Tis / 5% H2O / 5% EDT / 2.5% anisole thioether + NH4I (150 mg / ml)), and lyse at room temperature (25℃) for 2.5 h. Filter. At 0℃, add 10V MTBE to the filtrate; solid precipitates. Centrifuge, wash the solid twice with MTBE, and dry the centrifuged solid with N2 to obtain crude peptide Pep01-2, which can be used directly in the next reaction without further purification.

[0972] 1.4 Disulfide bond cyclization:

[0973] Weigh the crude peptide Pep01-2 and add it to a reaction flask. Dissolve it in a 10%-20% ACN / water solution to prepare a solution with a concentration of 0.5 mg / mL-1 mg / mL. Add 0.2 mmol / mL I2 / MeOH solution dropwise until the reaction solution turns pale yellow. Incubate the system at room temperature (25℃) for 10-15 min. Add saturated sodium vitamin C aqueous solution dropwise until the pale yellow color of the reaction solution disappears. The system is then purified by high-performance liquid chromatography and transsalted to obtain peptide Pep01.

[0974] Salt conversion conditions:

[0975] After high efficiency preparative liquid chromatography purification, the sample was diluted and pumped into the chromatographic column. After equilibration with 95% C mobile phase for 3 column volumes, it was then equilibrated with 95% A mobile phase for 3 column volumes. The above gradient was then used for preparation, and the fraction was freeze-dried to obtain the product.

[0976] MS m / z (ESI): 974.5 [(M+2H) / 2] + 650.3[(M+3H) / 3] + .

[0977] Starting from the corresponding starting materials, the following polypeptides were synthesized following the steps described above:

[0978] Synthetic route fifteen:

[0979] Synthesis of polypeptide intermediates:

[0980] Chemical solid-phase synthesis:

[0981] 1.1 Composite Table:

[0982] 1.2 Operating Steps:

[0983] 1.2.1 Weigh 1g of Rink amide-MBHA resin (degree of substitution 0.343mmol / g) and add it to the reactor. Swell the resin with 10V dichloromethane for 30min. After filtration, wash the resin twice with 10V DMF.

[0984] 1.2.2 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0985] 1.2.3 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive.

[0986] 1.2.4 Weigh 3.0 eq Fmoc-Lys(Alloc)-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0987] 1.2.5 After the reaction was completed, the ninhydrin / tetrachlorobenzoquinone test was negative; after filtration, the resin was washed 5 times with 10V DMF.

[0988] 1.2.6 Repeat steps 1.2.2-1.2.5 according to the synthesis table to condense amino acids 2-14 in sequence.

[0989] 1.2.7 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0990] 1.2.8 Weigh 10.0 eq Ac2O, 20.0 eq DIEA, and 10 V DMF and add them to the reactor. Purge with N2 for 1 h.

[0991] 1.2.9 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0992] 1.2.10 Add 20V DCM, 20eq PhSiH3, 0.2eq Pd(PPh4)3, and N2 to the reactor and purge for 20 min; repeat the operation twice; then wash the resin 6 times with 10V DMF to obtain intermediate Int-P1-2.

[0993] 1.2.11 Weigh 3.0 eq Fmoc-AEEA-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0994] 1.2.12 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0995] 1.2.13 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[0996] 1.2.14 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive. The intermediate Int-P2-1 was obtained.

[0997] 1.2.15 Weigh 3.0 eq Fmoc-AEEA-OH, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF and add them to the reactor. Purge with N2 for 0.5 h.

[0998] 1.2.16 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[0999] 1.2.17 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[1000] 1.2.18 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive. The intermediate Int-P3-1 was obtained.

[1001] 1.2.19 Weigh 3.0 eq Fmoc-Glu-O t Bu, 3.0 eq Oxyma, 6.0 eq DIC, and 10 V DMF were added to the reactor, and N2 was blown in for 0.5 h.

[1002] 1.2.20 After the reaction was completed, the resin was washed 5 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was negative.

[1003] 1.2.21 Add 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min. After filtration, add another 10V 20% Pip / DMF to the reactor and purge with N2 for 20 min.

[1004] 1.2.22 After the reaction was completed, the resin was washed 6 times with 10V DMF. The ninhydrin / tetrachlorobenzoquinone test was positive. The intermediate Int-P4-1 was obtained.

[1005] The polypeptide intermediate obtained above was washed twice with MeOH and twice with DCM. The resin was then dried to a fine sand-like state.

[1006] 1.3 Cutting and pyrolysis:

[1007] The peptide intermediates Int-P1-2, Int-P2-1, Int-P3-1, and Int-P4-1 were weighed separately and placed into centrifuge tubes. 10V cleavage buffer (95% TFA / 2% Tis / 2% H2O / 1% anisole thioether) was added, and the mixture was lysed at room temperature for 2.5 h. The filtrate was then filtered. At 0℃, 10V MTBE was added to the filtrate, causing solid precipitation. The solid was centrifuged, washed twice with MTBE, and dissolved in acetonitrile. The solid was then purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain crude peptides Int-P1-3, Int-P2-2, Int-P3-2, and Int-P4-2.

[1008] 1.4 Disulfide bond cyclization:

[1009] Crude peptides Int-P1-3, Int-P2-2, Int-P3-2, and Int-P4-2 were weighed and added to a reaction flask, dissolved in 50% methanol / water solution to prepare a 40 mg / mL solution. 1.5 eq I2 / MeOH solution was added dropwise until the reaction solution turned pale yellow. The system was reacted at room temperature (25℃) for 10-15 min. Saturated sodium vitamin C aqueous solution was added dropwise until the pale yellow color disappeared. The system was purified by C18 reversed-phase medium-pressure column chromatography (acetonitrile / water (0.5% trifluoroacetic acid)) to obtain compounds Int-P1, Int-P2, Int-P3, and Int-P4.

[1010] Int-P1-3: MS m / z(ESI): 1040.2[(M+2H) / 2] + ;

[1011] Int-P1: MS m / z(ESI): 1038.7[(M+2H) / 2] + ;

[1012] Int-P2-2: MS m / z(ESI): 1112.3[(M+2H) / 2] + ;

[1013] Int-P2: MS m / z(ESI): 1111.0[(M+2H) / 2] + ;

[1014] Int-P3-2: MS m / z(ESI): 1184.7[(M+2H) / 2] + ;

[1015] Int-P3: MS m / z(ESI): 1184.1[(M+2H) / 2] + ;

[1016] Int-P4-2: MS m / z(ESI): 1249.9[(M+2H) / 2] + ;

[1017] Int-P4: MS m / z(ESI): 1248.5[(M+2H) / 2] + .

[1018] Biological Experiment Examples

[1019] Experimental Example 1: Determination of the binding affinity of a compound to human IL-23R protein using surface plasmon resonance (SPR) method

[1020] 1. Experimental Objective

[1021] The affinity of the compound for human IL-23R protein was determined using surface plasmon resonance technology with a Biacore instrument.

[1022] 2. Experimental materials and equipment

[1023] 2.1 Instruments and Equipment

[1024] Instrument Name: Biacore; Model: T200; Equipment Number: M239; Supplier: Cytiva.

[1025] 2.2 Sample Information

[1026] Sample name: Human IL-23R, hFc Tag; Product number: ILR-HM223.

[1027] 2.3 Chips and Reagents

[1028] Table 1

[1029] 3. Experimental Methods

[1030] 3.1 Preparation of buffer solutions

[1031] 1) 1xHBS-EP +Preparation of (10mM HEPES, 150mM NaCl, 3mM EDTA, 0.05% surfactant P20, pH 7.4): Add 100mL of 10×HBS-EPES... + A solution of (0.1M HEPES, 1.5M NaCl, 0.03M EDTA, 0.5% surfactant P20, pH 7.4) was added to 900mL of ultrapure water, mixed thoroughly, filtered through a 0.22μm membrane, and then evacuated.

[1032] 2) High-salt HBS-EP + Preparation of running buffer: Take 500 mL of 1×HBS-EP + Add 10.2375g of sodium chloride to the buffer solution.

[1033] 3.2 Pierce TM Protein A is coupled to the CM5 chip.

[1034] 1) Using 1×HBS-EP + The experimental run buffer consisted of 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% surfactant P20, pH 7.4. Channel 12 of the CM5 chip was selected, and a freshly mixed 1:1 mixture of 50 mM N-hydroxysuccinimide (NHS) and 200 mM 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC) was injected into channel 12 at a flow rate of 10 μL / min for 600 s.

[1035] 2) Pierce TM Protein A was diluted to 100 μg / ml with 10 mM Acetate 4.5 and coupled to channel 12 of the CM5 chip. The injection time was 600 s, and the coupling level was approximately 5200 RU.

[1036] 3) Inject 1M ethanolamine at a flow rate of 10 μL / min for 300 s to block the remaining activation sites. Equilibrate the chip with the coupled protein at a flow rate of 10 μL / min.

[1037] 3.3 Affinity determination of the compound with Human IL-23R,hFc Tag protein

[1038] 1) Using 1×HBS-EP +The experimental buffer (10 mM HEPES, 500 mM NaCl, 3 mM EDTA, 0.05% surfactant P20, pH 7.4) was used to determine the binding and dissociation of one concentration of the compound with Human IL-23R and hFc Tag proteins in each injection cycle. Channel 1 served as a blank reference channel.

[1039] 2) Human IL-23R,hFc Tag protein, diluted to a concentration of 20 μg / ml, was captured into channel 2 of the CM5 chip at a flow rate of 10 μL / min for 120 s, with a capture level of approximately 2200 RU.

[1040] 3) Then inject compound samples of different concentrations (0.625-10 nM, 2-fold dilution, the specific concentration shall be subject to the actual concentration) at a flow rate of 50 μL / min, and record the binding and dissociation process of each compound with Human IL-23R and hFc Tag protein. The binding time is 120 s and the dissociation time is 600 s.

[1041] Chip regeneration was achieved by injecting 10 mM Glycine 1.5 at a flow rate of 30 μL / min for 30 s. The experimental temperature was 25 °C.

[1042] 4. Data Analysis

[1043] The affinity between the compound and human IL-23R protein was analyzed using the T200 analysis software (version 3.2) in kinetic analysis mode 1:1 binding, where k a k is the binding rate constant. d K is the dissociation rate constant. D is the affinity constant.

[1044] 5. Experimental Results

[1045] The experimental results are shown in Table 2 below.

[1046] Table 2

[1047] The results showed that the compounds of the present invention, including Pep01, Pep02, Pep06, Pep09, Pep03, Pep04, Pep05, Pep07, and Pep08, exhibited high binding affinity and stable binding characteristics. The following experimental examples demonstrate that the compounds of the present invention have good biological effects and in vivo pharmacokinetic properties in in vitro and in vivo experiments of IL-23R-related diseases.

[1048] The structure of Ref A is shown below:

[1049] Ref A is the second compound on page 147 of the specification in WO2024155553, and was prepared according to the preparation method disclosed in WO2024155553.

[1050] Experiment Example 2: PBMC Detection of pSTAT3

[1051] 1. Experimental objective:

[1052] The inhibitory effect of the compound on pSTAT3 expression in PBMCs was evaluated using ELISA.

[1053] 2. Relevant samples:

[1054] compound

[1055] 3. Cells:

[1056] Normal human PBMC cells

[1057] 4. Experimental Procedure:

[1058] Cryopreserved PBMCs were revived and stimulated for 3 days with 5 μg / mL of anti-human CD3 (Thermo, 16-0037-38) and 1 μg / mL of anti-human CD28 (Thermo, 16-0289-85). Cells were collected and induced again for 2 days under the same conditions, for a total of 5 days. Cells were collected, resuspended in serum-free medium, and seeded in 96-well plates for overnight incubation. Different concentrations of the compounds were added, and the cells were incubated at 37°C and 5% CO2 for 1 hour. Then, human IL-23 (biotechne, A10123091) was added to a final concentration of 20 ng / mL, and the cells were incubated at 37°C and 5% CO2 for 30 minutes. Lysis buffer (PhosStop: 10X; cycloplegic rehydration solution) was added. TM Cells were lysed using a 50X Cocktail, 10X Lysis Buffer, and 100X PMSF. pSTAT3 was measured according to the ELISA instructions (CST, 7300°C), and the absorbance at OD450 was read. IC50 was calculated using a nonlinear fitting four-parameter formula in GraphPad Prism 8 software. 50 Fitting.

[1059] 5. Results:

[1060] The experimental results are shown in Table 3 below.

[1061] Table 3

[1062] Ref B is compound 1-16-1 in WO2024114762, which was prepared according to the preparation method disclosed in WO2024114762.

[1063] Experimental Example 3: pSTAT3 Inhibition Test of Normal Human PBMCs (with 2% HSA)

[1064] 1. Experimental objective:

[1065] The compound was tested for its inhibitory effect on IL-23-induced pSTAT3 in normal human PBMC cells.

[1066] 2. Relevant samples:

[1067] compound

[1068] 3. Cells:

[1069] Normal human PBMC cells (purchased from Shanghai Junxing Biotechnology Co., Ltd.)

[1070] 4. Main reagents and consumables:

[1071] RPMI 1640 medium (ATCC modified) (Peiyuan Biotechnology, L240KJ), fetal bovine serum (EcoSai Biotechnology, FND500), PBS (Yuanpei, B310KJ), anti-CD3 (BioLegend, 317347), IL-1β (Acrobiosystems, ILB-H4110), 96-well plate (Corning, 3799), Human IL-23 alpha & IL-12 beta Heterodimer Protein, His Tag & Tag Free (MALS verified) (Acrobiosystems, ILB-H52W5-50ug), BSA (Fatty Acid & IgG Free, BioPremium) (Beyotime, ST025-20g), human serum albumin (HSA, Sigma-Aldrich, S#A1653-10G), protease phosphatase inhibitor mixture (general type, 50X) (Beyotime, P1045). Phospho-Stat3(Tyr705)Sandwich ELISA Kit#7300(CST, 7300C)

[1072] 5. Experimental Procedure:

[1073] 1) Day 1 induces PBMC

[1074] Prepare 5 μg / ml anti-CD3 with PBS, add 6 ml to T75 Flask, and incubate at 37°C for 2 hours. Resuscitate PBMCs with complete medium (1640 medium + 10% heat-inactivated FBS + 1X P / S), centrifuge at 400g for 10 min, resuspend in induction medium (1640 medium + 10% heat-inactivated FBS + 1X P / S + 100 ng / ml IL-1β) and count the cells. Then prepare a 0.5 M / ml cell suspension with induction medium. Wash the pre-coated T75 cells three times with PBS, add 25 ml of 0.5 M / ml cell suspension, and incubate at 37°C for 72 hours.

[1075] 2) Day 4-Day 5: Compound and cytokine treatment, ELISA detection

[1076] PBMCs were collected by centrifugation at 400g for 10 min, washed twice with starvation medium (1640 medium + 0.1% BSA), resuspended in starvation medium, counted, and prepared to a 4M / ml cell suspension. The cells were then transferred to 6-well plates and starved at 37°C for 4 hours. Subsequently, the cells were washed once by centrifugation at 400g for 5 min with 5 ml of 2% HSA (1640 medium + 0.1% BSA + 2% HSA), resuspended in 2% HSA 1640, counted, and prepared to a 2M / ml cell suspension with 2% HSA 1640. 100 μl of the cell suspension was added to each well. 4X compound was prepared using 2% HSA 1640, with a top concentration of 20 nM, and serially diluted 4-fold to obtain 8 concentration points (excluding the 0 point). 50 μl was added to each well, mixed, and incubated at 37°C for 30 min. 4X IL-23 (20 ng / ml, 400 pM) was prepared using 2% HSA 1640. Add 50 μL to each well, mix well, and incubate at 37°C for 30 min. Centrifuge at 1200 rpm for 2 min and discard the supernatant. Add 250 μL of pre-chilled PBS to each well, centrifuge at 1200 rpm for 2 min, and discard the supernatant. Add 120 μL of pre-chilled 1.2X Lysis buffer (CST, 7300C comes with Lysis buffer; 1X protease phosphatase inhibitor should be prepared fresh for use) to each well, mix thoroughly, and incubate at 4°C with shaking at 500 rpm for 30 min. Centrifuge at 4°C, 4000 rpm for 10 min, and then aspirate 100 μL of the supernatant into each well to an ELISA plate (CST, 7300C). Seal the plate and incubate overnight at 4°C. Perform ELISA assays the next day.

[1077] 6. Results and Discussion

[1078] The experimental results are shown in Table 4 below:

[1079] Table 4

[1080] The structure of Ref C is shown below:

[1081] Ref C is compound 7 in WO2024155552 (pages 301-303 of the specification), prepared according to the preparation method disclosed in WO2024155552.

[1082] Experimental Example 4: Inhibition of IFNγ secretion from normal human NK cells (with 2% HSA)

[1083] 1. Experimental objective:

[1084] The inhibitory effect of the compound on IFNγ secretion in normal human NK cells was tested.

[1085] 2. Relevant samples:

[1086] compound

[1087] 3. Cells:

[1088] Normal human NK cells (Frozen CD56+ Natural Killer Cells, purchased from Shanghai Aoneng Biotechnology Co., Ltd.)

[1089] 4. Main reagents and consumables:

[1090] NK-IV medium (Sanyi Hematopoietic Cell Propagation, CT-001), Human IL-18 (MCE, HY-P70591), Human IL-23alpha & IL-12 beta Protein (Kactus, IL2-HM1AB-100μg), Human serum albumin (HSA, Sigma-Aldrich, S#A1653-10G), Human IFN-γ Precoated ELISA Kit (Dakowei, 1110003), 96-well plate (Corning, 3799)

[1091] 5. Experimental Procedure:

[1092] 1) Day 1: Treatment of cells with compounds and cytokines

[1093] 4X compound was prepared using NK-IV medium (containing 2% HSA) with a top concentration of 40 nM and serially diluted 4-fold to obtain 9 concentration points (excluding the 0 point). NK cells were resuscitated, resuspended in NK-IV medium (containing 2% HSA), and counted. Then, a 0.45 M / ml cell suspension was prepared using NK-IV medium (containing 2% HSA), and 100 μL of cell suspension was added to each well. Next, 50 μL of 4X compound was added to each well, mixed, and incubated at 37°C for 30 min. Finally, 4X cytokines (IL-23 100 ng / ml and IL-18 200 ng / ml) were prepared using NK-IV medium (containing 2% HSA), and 50 μL of 4X cytokines was added to each well, mixed, and incubated overnight at 37°C.

[1094] 2) Day 2 ELISA test

[1095] After centrifuging at 300g for 10 minutes, take 100ul of the supernatant and add it to the ELISA plate to complete the ELISA detection.

[1096] 6. Results and Discussion

[1097] The experimental results are shown in Table 5 below:

[1098] Table 5

[1099] Conclusion: The compounds of this invention exhibit high activity in the albumin-containing test system.

[1100] Experimental Example 5: Rat Splenic Cell IL17A Secretion Inhibition Assay

[1101] 1. Experimental Objective

[1102] The ability of the compound to inhibit IL-23-induced interleukin-17A (IL-17A) production in rat spleen cells was determined by ELISA to assess the compound's activity in inhibiting the IL23 / IL-23R signaling pathway.

[1103] 2. Main Reagents and Instruments

[1104] 2.1 Main Reagents

[1105] Table 6

[1106] 2.2 Main Instruments

[1107] Name: Multifunctional Microplate Reader; Brand: Molecular Devices; Model: i3x.

[1108] 3. Experimental methods and procedures

[1109] 1) Preparation of cell culture medium

[1110] A complete cell culture medium was prepared, consisting of 90% RMPI-1640, 10% heat-inactivated rat serum, and 1% penicillin-streptomycin solution.

[1111] 2) Preparation of compound solutions

[1112] The compound was serially diluted. The initial concentration of the compound was 48.8 nM, and it was serially diluted 4-fold, resulting in a total of 8 concentration gradients.

[1113] 3) Preparation of single-cell suspension of rat spleen cells

[1114] After euthanizing rats with carbon dioxide, the spleen was immediately removed and placed on a 70 μm cell sieve in a 6-well plate containing PBS. The spleen tissue was gently ground with a syringe plunger to prepare a single-cell suspension.

[1115] 4) Red blood cell lysis

[1116] Collect the cell suspension and centrifuge at 1800 rpm for 5 minutes. Discard the supernatant, resuspend the cell pellet in red blood cell lysis buffer, and incubate at room temperature for 5 minutes.

[1117] 5) Cell washing

[1118] Add an equal volume of PBS to terminate the lysis reaction, centrifuge at 1800 rpm for 5 minutes, and discard the supernatant. Resuspend the cells in PBS and wash them once more.

[1119] 6) Preparation of stimulation culture medium

[1120] Rat IL-23 and rat IL-1β were added to complete cell culture medium to a final concentration of 30 ng / mL, which was then used as a conditioned medium for cell stimulation.

[1121] 7) Cell counting and resuspension

[1122] Resuspend cells in conditioned medium, count them, and adjust the cell density to 6 × 10⁶. 6 per mL.

[1123] 8) Cell plating and compound treatment

[1124] Add 100 μL of cell suspension and 100 μL of each concentration of compound solution to each well of a 96-well plate, and incubate in a CO2 incubator for 3 days.

[1125] 9) Sample collection

[1126] After 3 days of culture, 100 μL of supernatant was aspirated from each well for subsequent ELISA detection of rat IL-17A.

[1127] 10) ELISA testing and data analysis

[1128] The rat IL-17A ELISA kit was tested and analyzed according to the manufacturer's instructions. Data analysis was performed using GraphPad Prism software (version 9.5) with four-parameter nonlinear regression fitting to calculate the IC50. 50 value.

[1129] result

[1130] The experimental data are shown in Table 7 below.

[1131] Table 7

[1132] Experiment Example 6: Rat Pharmacokinetic Experiment:

[1133] This experiment evaluated the in vivo pharmacokinetic properties of rats via intravenous injection, oral administration, and duodenal intubation.

[1134] Experimental methods and conditions: Male SD rats were administered the test compound once orally at doses of 1 mg / kg (intravenous injection (IV), solvent: PBS), 10 mg / kg (oral administration (PO), solvent: PBS + 100 mg / kg penetration enhancer NaClO), and 10 mg / kg (intraduodenal cannulation administration (ID), solvent: PBS + 100 mg / kg penetration enhancer NaClO). Blood samples were collected via the jugular vein at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after administration. Approximately 0.2 mL of each sample was collected. The plasma was anticoagulated with K2-EDTA and centrifuged for analysis. Plasma drug concentration was determined using liquid chromatography-tandem mass spectrometry (LC / MS / MS) to calculate pharmacokinetic parameters. The results are shown in Tables 8 and 9 below.

[1135] Table 8. Pharmacokinetics of ID (10 mg / Kg)

[1136] Table 9. Pharmacokinetics of PO administration (10 mg / Kg)

[1137] Conclusion: Compared with the clinical molecule JNJ-2113, the compound of this invention has pharmacokinetic advantages in rats, including a significantly prolonged half-life and greatly increased plasma exposure.

[1138] Experiment Example 7: Pharmacokinetic Experiment of Crab-Eating Monkeys:

[1139] This experiment evaluated the in vivo pharmacokinetics of cynomolgus monkeys after oral administration.

[1140] Experimental methods and conditions: Male cynomolgus monkeys were administered the test compound once. Blood samples were collected via peripheral vein at 5 min, 30 min, 2 h, 8 h, 24 h, 48 h, 96 h, and 168 h after administration. Approximately 0.5 mL of blood was collected from each sample. The plasma was anticoagulated with K2-EDTA and centrifuged for analysis. Plasma drug concentration was determined using liquid chromatography-tandem mass spectrometry (LC / MS / MS) to calculate pharmacokinetic parameters. The results are shown in Table 10 below.

[1141] Table 10. Pharmacokinetics of po administration

[1142] Conclusion: Compared with the clinical molecule JNJ-2113 and the published patented molecules Ref A, B, the compounds of this invention have pharmacokinetic advantages in cynomolgus monkeys, including a significantly prolonged half-life and greatly increased plasma exposure.

[1143] Experiment Example 8: In vivo efficacy experiment of IL23-induced rat otitis model

[1144] 1. Experimental Objective

[1145] The purpose of this experiment was to evaluate the efficacy of the compound in a rat model of psoriasis induced by human interleukin-23.

[1146] 2. Experimental Materials

[1147] 2.1 Reagents and Materials

[1148] hIL-23 (ILB-H52W5, Acrobiosystem), phosphate buffer (pH: 7.2) (Gibco), phosphate buffer (pH: 7.4) (Gibco), penetration enhancer

[1149] 2.2 Instruments and Equipment

[1150] Electronic balance, electronic scale, centrifuge, ultrasonic cleaner, electronic vibrator, anesthesia machine

[1151] 2.3 Animal Information

[1152] Table 11

[1153] Note: All animal handling procedures in this experimental protocol were approved by the International Agency for Animal Welfare and Use (IACUC).

[1154] 3. Experimental Methods

[1155] 3.1 Grouping

[1156] After the adaptation period, all rats will be randomly grouped according to their body weight.

[1157] 3.2 Model Establishment

[1158] Except for the blank control group, rats in all other groups were injected intradermally with 2.5 μg of human interleukin-23 in the right ear to induce a rat psoriasis model. The injection was administered once daily on days 0, 1, 2, and 3, for a total of four injections. Each injection was a single-point injection with a volume of 20 μL. The blank control group received an equal volume of phosphate-buffered saline injected intradermally in the right ear as a control.

[1159] 3.3 Drug administration

[1160] All rats were administered the compound or solvent (model group) from day -1 to day 3 for a total of 5 days. The compound was administered daily 30 minutes before the injection of human interleukin-23. The group receiving the compound twice daily had a 10-hour interval between administrations.

[1161] 3.4 Data Processing

[1162] 1) Ear thickness measurement

[1163] Ear thickness was measured daily in all animals from day -1 to day 4, for a total of 6 times. The initial ear thickness (ear thickening) was subtracted from the daily ear thickness in each group and compared with the model group. Ear thickness results from day 4 to day -1 (the last day of the experiment) were analyzed using GraphPad Prism. Ear thickening inhibition rate % = [1 - (mean of the treated group / mean of the model group)] × 100%.

[1164] 2) PK test samples (day 3)

[1165] Animal plasma was collected for PK testing (baseline, 0.25h, 2h, 6h, 24h). Approximately 0.2 mL of whole blood was collected from rats via the jugular vein and added to a tube containing K2-EDTA as an anticoagulant. The tube was placed on wet ice until centrifugation. Samples were centrifuged (3200×g, 4℃, 10 min) within 1 hour of collection. Collected plasma samples were stored at -80℃ until bioanalysis.

[1166] 4. Experimental endpoint

[1167] On the final day of the experiment (day 4), all rats were euthanized with carbon dioxide. Right ears were collected, processed, weighed, and then processed with RNA Later overnight. The supernatant was discarded the next day, and the ear samples were stored at -80°C. Samples were collected for Q-PCR analysis of IL17A relative expression levels.

[1168] 5. Q-PCR analysis method

[1169] 5.1 Experimental Materials

[1170] 5.1.1 Reagents

[1171] TRIzol TM Reagents: Invitrogen, catalog number: 15596018; PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time): Takara, catalog number: RR047A; TB Premix Ex Taq TM II (Tli RNaseH Plus): Takara, catalog number: RR820A; qPCR primers (see Table 12), Suzhou Genewiz.

[1172] Table 12

[1173] 5.1.2 Instruments

[1174] Tissue homogenizer: Shanghai Jingxin, model Tissuelyser-24L; Spectrophotometer: Thermo Scientific, model Nanodrop2000; PCR thermal cycler: Applied Biosystems, model 4375305; Real-time PCR instrument: Bio-Rad, model CFX384.

[1175] 5.2 PCR detection method

[1176] 5.2.1 RNA Extraction

[1177] 1) Add 1 ml of TRIzol lysis buffer to every 20-30 mg of tissue and homogenize using a pre-cooled tissue homogenizer (60 Hz, 1 minute, 2 times).

[1178] 2) Incubate for 5 minutes to allow the nucleoprotein complex to completely dissociate, centrifuge at 12000×g for 5 minutes at 4°C, and transfer the supernatant to a new centrifuge tube.

[1179] 3) Add 0.2 mL of chloroform, mix thoroughly, and let stand at room temperature for 2-5 minutes.

[1180] 4) Centrifuge at 12000×g for 15 minutes at 4℃. The mixture separates into a lower red phenol-chloroform phase, a middle phase, and a colorless upper aqueous phase. Transfer the aqueous phase containing RNA to a new test tube.

[1181] 5) Add 0.5 mL of isopropanol to the aqueous phase, mix gently, and let stand at -20°C for 1 hour or overnight.

[1182] 6) Centrifuge at 4℃, 12000×g for 15 minutes. Total RNA will precipitate at the bottom of the test tube, forming a white gel-like precipitate. Discard the supernatant.

[1183] 7) Resuspend the precipitate in 1 mL of 75% ethanol. Vortex the sample briefly, then centrifuge at 7500 × g for 5 minutes at 4 °C. Discard the supernatant.

[1184] 8) Repeat step 7.

[1185] 9) Vacuum or air dry the RNA particles for 5-10 minutes.

[1186] 10) Dissolve the RNA precipitate in 40 μL of enzyme-free water and store at -80℃ for a long time.

[1187] 11) The 260 / 280 ratio was determined using Nanodrop 2000.

[1188] 5.2.2 Reverse transcription reaction

[1189] Genomic DNA elimination reaction

[1190] 1) Prepare the genomic DNA elimination reaction solution on ice (Table 13).

[1191] 2) Set the program on the PCR instrument to 42℃ for 2 minutes, then store at 4℃.

[1192] Table 13 Genomic DNA Elimination Reagents

[1193] Reverse transcription reaction

[1194] 1) Prepare the reverse transcription reaction solution on ice (see Table 14 for reverse transcription reaction reagents). Add 10 μL of the reaction solution from the previous step to the prepared solution, mix gently, and immediately proceed with the reverse transcription reaction.

[1195] 2) Set the program on the PCR instrument to 37℃ for 15 minutes, 85℃ for 5 seconds, and store at 4℃.

[1196] 3) After the reaction is complete, the cDNA should be stored at -20℃ for a long period of time.

[1197] Table 14 Reverse Transcription Reagents

[1198] 5.2.3 qPCR reaction

[1199] 1) Prepare qPCR reaction solutions (qPCR reaction reagents, Table 15) on ice.

[1200] 2) Add the 384-well plate to the Real-Time PCR instrument, set the PCR program according to Table 16 and start the reaction immediately.

[1201] 3) Calculate the expression of the target gene using the ΔΔCt method.

[1202] Table 15 qPCR Reagents

[1203] Table 16 PCR Procedure

[1204] 6. Data Analysis

[1205] Experimental data are expressed as mean ± standard error (Mean ± SEM) and analyzed using GraphPad Prism software with ANOVA, one-way ANOVA and two-way ANOVA methods (*P<0.05, **P<0.01, ***P<0.001; ****P<0.0001).

[1206] 7. Experimental Results

[1207] The percentage of ear thickening inhibition and the relative expression level of IL17A are shown in Table 17 below.

[1208] Table 17

[1209] Conclusion: The preferred compound of this invention can effectively inhibit ear thickening in an in vivo pharmacological experiment of an IL23-induced rat otitis model; when administered once daily, it has a more significant effect on inhibiting ear thickening than the clinical molecule JNJ-2113 and the patented molecules Ref B and Ref C at the same or even higher doses.

[1210] In an in vivo pharmacological experiment of the preferred compound of this invention in an IL23-induced rat otitis media model, it effectively inhibited the expression of IL17A downstream of the signaling pathway. When administered once a day, it showed a more significant inhibitory effect on IL17A expression than the clinical molecule JNJ-2113 and the patented molecule Ref B at the same or even higher doses.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, c[X4-X5-X6-X7-X8-X9]-X10-X11-X12-X13-X14-X15-X16 (I) in, In c[X4-X5-X6-X7-X8-X9], "c[]" indicates that X4 and X9 are connected in a loop; X6, X7, X8, X11, X12, X13, and X14 are any amino acid residues; X4 is The carbonyl end is connected to X5; R 4 -C(=O)R 4a , -L 1 -Z 1 or -L 2 -(L 3 ) n1 -(L 4 ) n1a -Z 2 ; R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl; X5 is The carbonyl end is connected to X6; R 5a C 1-6 Alkyl, -L 1 -Z 1 Or be -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 alkyl; R 5a-1 and R 5a-2 Independently H or C 1-6 Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups; R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl; X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8; R 9a and R 9b Independently H or C 1-6 alkyl; X10 is The carbonyl end is connected to X11; R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy; R 10a-1 and R 10a-2 Independently -NH2, -OH, or halogen; Cycle B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or a ring with one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups; R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen; X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl; X16 is R 16 -NH2, -L 1 -Z 1 -L 2 -(L 3 )n1-L 4 -Z 2 or -L 2 -(L 3 ) n1 -(L 4 ) n1a -Z 2 ; And at least one of X4 and X16 contains -L 2 -(L 3 ) n1 -L 4 -Z 2 or -L 2 -(L 3 )n1-(L 4 ) n1a -Z 2 structure; L 1 and L 2 Independent of chemical bonds or connecting units; Z 1 Independently Z 1a and Z 1c Independently -NH-, -C(=O)-, -O-, or -S-; Z 1b H, -SF5, halogen, C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl group, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl; Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl; L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -N(R L3 - or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -N(R L3 )-C(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 Alkylene-N(R) L3 )-; R L3 For H or C 1-6 alkyl; L 4 Independently R L4a and R L4b H and C independently 1-6 alkyl or Z 1g H, halogen or C 1-6 alkyl; Z 2 Independently -C (=O) -C 6-24 Alkylene-R z ;R z -CO2H, -SO3H, n1, n1a, n3, n4, n5, n6, n7, n8, n9, n9a, n9b, n10, n10a, n11, n12 and n13 are independently 0, 1, 2, 3, 4, 5 or 6; In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3. The condition is that the compound represented by formula (I) satisfies one, two, or three of the following conditions: (1) X10 is (2)X15 is (3) X8 is 2. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) is the same as the compound shown in formula (I-5): X4 is The carbonyl end is connected to X5; X5 is The carbonyl end is connected to X6; X6 is The carbonyl end is connected to an amino group; X8 is The carbonyl end is connected to an amino group; X12 is The carbonyl end is connected to an amino group; X10 is The carbonyl terminus is connected to an amino group; ring B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or is bonded to one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups; R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2; X15 is The carbonyl end is connected to X16; ring A is a 5-6 membered heteroaryl group, which is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl; X16 is R 16 -L 216 -(L 316 )n1-(L 416 ) n1a -Z 216 ; L 216 -NH-C 1-6 alkylene-NH- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H, n1 and n1a are independently 0, 1 or 2; n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5; m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5; m13 can be 0, 1, 2, 3, 4 or 5 independently; In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is one, two or three.

3. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) is the same as the compound shown in formula (Ih): Wherein, ring B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cyclic alkenyl group, or is surrounded by one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups; R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2; Ring A is a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl; X16 is R 16 -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ; L 216 -NH-C 1-6 alkylene-NH- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H, n1 and n1a are independently 0, 1 or 2; n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5; m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5; m13 can be 0, 1, 2, 3, 4 or 5 independently; In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3. Preferably, for for 4. The compound of formula (I) as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1) When the compound shown in formula (I) contains a cationic fragment, the anion is acetate, trifluoroacetate, chloride, bromide, adipic acid, benzoate, benzenesulfonate, citrate, decanoate, lactate, maleate, methanesulfonate, propionate, oxalate, succinate, sulfate, or tartrate; the cation and anion have equal charges. (2)L 1 and L 2 In this context, the connecting unit is a combination of one or more groups selected from the following: C 1-6 Alkylene, -C(=O)-, -NR La -、 -C(=O)NR Ld -、-NR Le C(=O)-、-C(=O)O-、-O-、-S-、-(CH2CH2O) m -、 Or by one or more R Lf Replacement C 1-6 Alkylene; R La R Lb R Lc R Ld and R Le Independently H or C 1-6 alkyl; R Lf C 1-6 Alkyl groups, -NHC(=NH)NH2, or -CONH2; m is 1, 2, 3, 4, 5, 6, 7 or 8; (3) X6, X7, X8, X11, X12, X13, and X14 are independently... R a For H or C 1-6 alkyl; R b -(CH2) p1 -R b-1 ; Or, R a R b Together with the carbon atoms it is attached to, they form C 3-8 Cycloalkyl, 4-10 membered heterocyclic alkyl, with one or more R b-2 Replacement C 3-8 cycloalkyl, or by one or more R b-3 Substituted 4-10 membered heterocyclic alkyl groups; R b-1 -OH, -CO2H, -CON(R) b-4 Rb -5 ), -N(R b-6 )C(=O)R b-7 C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 6-10 aryl, 5-10 membered heteroaryl, 4-10 membered heterocyclic alkyl, C substituted with one or more hydroxyl groups 1-6 Alkyl, with one or more R b-8 Replacement C 6- 10 aryl, with one or more R b-9 Substituted 5-10 heteroaryl groups, or substituted with one or more R groups b-10 Substituted 4-10 membered heterocyclic alkyl groups; R b-4 R b-5 and R b-6 Independently H or C 1-6 alkyl; R b-7 C 1-6 Alkyl or 4-10 membered heterocyclic alkyl; R b-2 R b-3 R b-8 R b-9 and R b-10 Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl; p1 is 0, 1, 2, 3, 4, 5 or 6; In the 4-10 membered heterocyclic alkyl and 5-10 membered heteroaryl groups, the heteroatom is one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; (4)X10 is The carbonyl end is connected to X11; R 10a C 1-6 Alkyl, C 1-6 Alkyl group, by one or more R 10a-1 Replacement C 1-6 Alkyl, or with one or more R 10a-2 Replacement C 1-6 Alkoxy; R 10a-1 and R 10a-2 Independently -NH2, -OH, or halogen; R 10b and R 10c H and C independently 1-6 Alkyl, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen; n2a is 0, 1, or 2; n2b and n2 are independently 1 or 2.

5. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1)L 1 Independent of chemical bonds, -C(=O)-C 1-6 Alkylene-C(=O)-, Or -C(=O)-(CH2CH2O) m2a -C 1-6 Alkylene NH- a a end and Z end 1 Connection; R Lb and R Lc Independently H or C 1-6 Alkyl group; m1, m2, m3 and m4 are independently 0, 1, 2, 3, 4 or 5; m2a is 1-10; (2)L 2 Independently for -L 2a -C 1-6 Alkylene-L 2b -、 -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 Alkylene- or -NH-G 1-6 Alkylene-C(=O)- b The C 1-6 Alkylene is optionally surrounded by one or more R Lf replace; L 2a and L 2b Independently -C(=O)- or -NH-; L 2c L 2d L 2e L 2f L 2g and L 2h Independently -C(=O)-, -CH2-, or -NH-; m is 1, 2, 3, 4 or 5; m5, m6, m7, m8, m9, m10, m11, m12, and m13 are independently 0, 1, 2, 3, 4, or 5; m14, m15 and m16 are independently 1, 2, 3, 4 or 5; R Lb and R Lc Independently H or C 1-6 alkyl; R Lf C 1-6 Alkyl groups, -NHC(=NH)NH2, or -CONH2; (3)L 3 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene -NH-; n1 is 0, 1 or 2; (4)L 4 Independently R L4a and R L4b H and C independently 1-6 alkyl or Z 1g H, halogen or C 1-6 alkyl; n10, n10a, n11, n12 and n13 are independently 1, 2, 3, 4 or 5.

6. The compound of formula (I) as claimed in claim 5, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1)L 1 Independent of chemical bonds, a-end and Z 1 connect; (2)L 2 Independent of chemical bonds, b end and L 3 connect; (3)Z 1 for (4)L 3 Independently c-end and L 4 Connection; n1 is 0, 1, or 2; (5)L 4 for d end and Z end 2 connect; (6)Z 2 for 7. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1) X4 is The carbonyl end is connected to X5; R 4 -C(=O)R 4a , -L 14 -Z 14 or -L 24 -(L 34 ) n1 -L 44 -Z 24 ; R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl; n1 is 0, 1, or 2; L 14 L as claimed in any one of claims 1-4 1 Definition; Z 14 Z as claimed in any one of claims 1-4 1 Definition; L 24 L as claimed in any one of claims 1-4 2 Definition; L 34 L as claimed in any one of claims 1-4 3 Definition; L 44 As defined in any one of claims 1-4; Z 24 Z as claimed in any one of claims 1-4 2 Definition; (2)X5 is The carbonyl end is connected to X6; R 5a C 1-6 Alkyl, -Z 15 Or be -CON(R) 5a-1 R 5a-2 ) replaced by C 1-6 Alkyl; R 5a-1 and R 5a-2 Independently H or C 1-6 Alkyl; or, R 5a-1 R 5a-2 Together with the nitrogen atom it is attached to, it forms a 4-10 membered heterocyclic alkyl group, or is formed by one or more R atoms. 5b Substituted 4-10 membered heterocyclic alkyl groups; R 5b Independently halogen, oxo group (=O), C 1-6 Alkyl, C 1-6 Halogenated alkyl or C 3-8 cycloalkyl; Z 15 for Z 1d Z 1e and Z 1f Independently H or C 1-6 alkyl; n8 can be 1, 2, 3, 4, or 5; (3)X10 is The carbonyl end is connected to X11; R 10a C replaced by -NH2 1-6 Alkoxy; R 10b and R 10c H and C independently 1-6 Alkyl, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2, -OH, or halogen; n2a is 0, 1, or 2; n2b and n2 are independently 1 or 2; (4)X15 is The carbonyl end is connected to X16; T 1 and T 4 Independently -NH-, -NR 15 -、-O- or -S-; T 2 T 3 T 5 and T 6 Independent of N, CH or CR 15 ;R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl; (5)X16 is R 16 -NH2, -L 116 -Z 116 or -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ; n1 and n1a are independently 0, 1 or 2; L 116 L as claimed in any one of claims 1-4 1 Definition of Z 116 Z as claimed in any one of claims 1-4 1 Definition; L 216 L as claimed in any one of claims 1-4 2 Definition; L 316 L as claimed in any one of claims 1-4 3 Definition; L 416 L as claimed in any one of claims 1-4 4 Definition; Z 216 Z as claimed in any one of claims 1-4 2 Definition; (6)X12 is R a R b Together with the carbon atoms it is attached to, they form C 3-8 Cycloalkyl, 4-10 membered heterocyclic alkyl, with one or more R b-2 Replacement C 3-8 cycloalkyl, or by one or more R b-3 Substituted 4-10 membered heterocyclic alkyl groups; R b-2 and R b-3 Independently halogen or C 1-6 alkyl.

8. The compound of formula (I) as claimed in claim 7, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1)L 14 For chemical bonds, -C(=O)-C 1-6 Alkylene-C(=O)-, Or -C(=O)-(CH2CH2O) m2a -C 1-6 Alkylene NH- a a end and Z end 14 Connection; R Lb and R Lc Independently H or C 1-6 alkyl; Z 14 for Z 1b It is a halogen; Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl; L 24 For chemical bonds, -C(=O)-C 1-6 alkylene-NH- b -C(=O)-C 1-6 Alkylene-C(=O)- b or The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 34 connect; R Lb and R Lc Independently H or C 1-6 alkyl; R Lf It is -NHC(=NH)NH2; L 34 -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 44 connect; L 44 for d end and Z end 24 connect; Z 24 -C(=O)-C 6-24 Alkylene -CO2H; n1 is 0, 1, or 2; n4, n5, n7, n8, n9, n10, and n11 are independently 1, 2, 3, 4, or 5; m2, m2a, m3, m4, m6, m7, m8 and m9 are independently 1, 2, 3, 4 or 5; (2)R 15 It can be -NH2, -CH2COOH, -CH2CH2COOH or -CH2CONH2; (3)L 116 It is a chemical bond; Z 116 for Z 1d Z 1e and Z 1f The site is H or C 1-6 alkyl; L 216 For chemical bonds, -NH-C 1-6 alkylene-NH- b , -NH-(CH2CH2O) m -C 1-6 Alkylene -C(=O)NH-(CH2CH2O) m -C 1-6 alkylene- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf For -CONH2; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H, R L4a and R L4b Independently for H or Z 1g It is a halogen; n1 is 0, 1, or 2; n7, n8, n9, n9a, n9b, n10, n10a, n11, n12 and n13 are independently 1, 2, 3, 4 or 5; m, m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5; m13 can be 0, 1, 2, 3, 4 or 5 independently.

9. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) satisfies one or more of the following conditions: (1) X4 is The carbonyl end is connected to X5; (2)X5 is The carbonyl end is connected to X6; (3)X6 is The carbonyl end is connected to X7; (4)X7 is The carbonyl end is connected to X8; (5)X8 is The carbonyl end is connected to X9; (6)X9 is The carbonyl end is connected to X10, and the nitrogen end is connected to X8; (7)X10 is The carbonyl end is connected to X11; (8)X11 is The carbonyl end is connected to X12; (9)X12 is The carbonyl end is connected to X13; (10)X13 is The carbonyl end is connected to X14; (11)X14 is The carbonyl end is connected to X15; (12)X15 is The carbonyl end is connected to X16; (13)X16 is 10. The compound of formula (I) as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound represented by formula (I) is a compound represented by formula (I-2), formula (I-3), or formula (I-4): The definitions of X4, X5, X8, X10, X15, X16, ring A, and ring B are as defined in any one of claims 1-7.

11. The compound of formula (I) as claimed in claim 1, characterized in that, The compound represented by formula (I) is a compound represented by formula (Ia), (Ib), (Ic), (Id), (Ie), (If), or (Ig): Among them, X5, X8, X10, X15, X16, R4, R 5a R 10b R 10c The definitions of n2a, n2b, and n2 are as defined in any one of claims 1-9; T 1 T 2 T 3 T 4 T 5 T 6 As defined in claim 7; Preferably, the compound represented by formula (Ib) satisfies any of the following conditions: (1)R 10b C 1-6 Alkyl group; n2 is 1 or 2; X5 is The carbonyl end is connected to an amino group; R 4 -C(=O)CH3, X16 is R 16 -L 216 -(L 316 ) n1 -L 416 -Z 216 ; L 216 -NH-C 1-6 alkylene-NH- b , The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene -CO2H; n1 is 0, 1, or 2; n9 and n10 are independently 1, 2, 3, 4 or 5; m6, m7, m8 and m9 are independently 1, 2, 3, 4 or 5; (2)R 10b C 1-6 Alkyl group; n2 is 1 or 2; X5 is The carbonyl end is connected to an amino group; X16 is X4 is The carbonyl end is connected to X5; R 4 -L 24 -(L 34 ) n1 -L 44 -Z 24 ; L 24 For chemical bonds, b end and L 34 connect; L 34 -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 44 connect; L 44 for d end and Z end 24 connect; Z 24 -C(=O)-C 6-24 Alkylene -CO2H; n1 is 0, 1, or 2; n9 and n10 are independently 1, 2, 3, 4 or 5; Preferably, in the compound represented by formula (Ie), X10 is... X5 is R 4 -C(=O)CH3, X16 is R 16 -L 216 -(L 316 ) n1 -L 416 -Z 216 ; L 216 -NH-C 1-6 alkylene-NH- b or The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene -CO2H; n1 is 0, 1, or 2; n9 and n10 are independently 1, 2 or 3; m6 and m7 are independently 1, 2, 3, 4 or 5; Preferably, in the compound shown in formula (Ig), R 4 -C(=O)CH3, X5 is The carbonyl end is connected to an X6 amino group; X6 is The carbonyl end is connected to an amino group; X8 is The carbonyl end is connected to an amino group; X12 is The carbonyl end is connected to an amino group; X10 is The carbonyl end is connected to an amino group; Cycle B is a 5-8 membered heterocyclic alkenyl group, a 5-8 membered cycloalkenyl group, or a ring with one or more R groups. 10-1 Substituted 5-8 membered heterocyclic alkenyl groups or those with one or more R groups 10-2 Substituted 5-8 membered cycloalkenyl groups; R 10-1 and R 10-2 Independently for C 1-6 Alkyl, oxo, or with one or more R 10b-1 Replacement C 1-6 Alkyl; R 10b-1 Independently -NH2; X15 is The carbonyl end is connected to X16; Ring A is a 5-6 membered heteroaryl group, wherein the 5-6 membered heteroaryl group is optionally surrounded by 1, 2 or 3 R groups. 15 Replace; R 15 Independently for C 1-6 Alkyl, -NR 15a R 15b Or by one or more R 15c Replacement C 1-6 Alkyl; R 15a and R 15b Independently H or C 1-6 Alkyl; R 15c Independently -CO2H, -CONH2, or -CO2C 1-6 alkyl; X16 is R 16 -L 216 -(L 316 ) n1 -(L 416 ) n1a -Z 216 ; L 216 -NH-C 1-6 alkylene-NH- b , or -NH-C 1-6 Alkylene-C(=O)- b The C 1-6 Alkylenes are optionally R Lf Replacement; b-end and L 316 Connection; R Lf -CONH2; R Lb and R Lc Independently H or C 1-6 alkyl; L 316 Independently -C (=O) -C 1-6 Alkylene-(OCH2CH2) n9 -NH- c -NH(CH2CH2O) n9 -C 1-6 Alkylene-C(=O)- c or -C(=O)-C 1-6 Alkylene-(OCH2CH2) n9a -NHC(=O)-(4-10 membered heterocyclic alkylene)-(CH2CH2O) n9b -C 1-6 alkylene-NH- c c end and L 416 connect; L 416 Independently d end and Z end 216 connect; Z 216 -C(=O)-C 6-24 Alkylene-R z ;R z -CO2H, -SO3H, n1 and n1a are independently 0, 1 or 2; n9, n9a, n9b, n10, n10a and n11 are independently 1, 2, 3, 4 or 5; m6, m7, m8, m9, m10, m11, m12, m14, m15 and m16 are independently 1, 2, 3, 4 or 5; m13 can be 0, 1, 2, 3, 4 or 5 independently; In the 5-6 membered heteroaryl, 5-8 membered heterocyclic alkenyl and 4-10 membered heterocyclic alkyl groups, the heteroatom is one, two or three of N, O and S, and the number of heteroatoms is 1, 2 or 3. More preferably, in the compound shown in formula (Ig), X10 is The carbonyl end is connected to an amino group; X15 is The carbonyl end is connected to an X16 amino group.

12. The compound of formula (I) as claimed in claim 1, characterized in that, The compound shown in formula (I) is any compound in Table A or Table B.

13. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-12 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutical excipient.

14. The use of a compound of formula (I) as described in any one of claims 1-12 or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 13, in the preparation of a medicament for the prevention and / or treatment of diseases associated with interleukin-23; wherein the diseases associated with interleukin-23 are preferably autoimmune diseases, such as psoriasis, arthritis, or inflammatory bowel disease.

15. A compound as shown in formula (II) or a pharmaceutically acceptable salt thereof, in, R 4 -C(=O)R 4a ;R 4a C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-8 cycloalkyl, or by one or more R 4b Replacement C 3-8 cycloalkyl; R 4b Independent of halogen, C 1-6 Alkyl or halogenated C 1-6 alkyl; X16 is R 16 It is -NH2; X5, X6, X8, X10 and X12 are as defined in claim 1, 4, 7 or 9; Preferably, the compound represented by formula (II) is any of the following compounds:

16. A compound as shown below, or a pharmaceutically acceptable salt thereof:

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