Peptide inhibitor of interleukin-23 receptor and use thereof

Abstract

The present invention relates to a peptide inhibitor of interleukin-23 receptor, and a use thereof. In particular, it relates to a peptide inhibitor of interleukin-23 receptor, a stereoisomer, pharmaceutically acceptable salt or solvate thereof, a pharmaceutical composition thereof, and a use of the peptide inhibitor in treating or preventing diseases or conditions comprising inflammatory bowel disease, Crohn's disease, and psoriasis.

Description

Peptide inhibitors of interleukin-23 receptor and uses thereof TECHNICAL FIELD

[0001] The present invention relates to novel peptide inhibitors of interleukin-23 receptor and their use in the treatment or prevention of a variety of diseases and conditions, including inflammatory bowel disease, Crohn's disease, and psoriasis. BACKGROUND

[0002] The interleukin-23 (IL-23) cytokine has been implicated in the pathogenesis of autoimmune diseases such as multiple sclerosis, asthma, rheumatoid arthritis, psoriasis, and inflammatory bowel disease (IBD). Model studies in mice of acute and chronic IBD show that IL-23R and downstream effector cytokines play a major role in disease pathogenesis. IL-23R is expressed on a variety of adaptive and innate immune cells, including Th17 cells, gd T cells, natural killer (NK) cells, dendritic cells, macrophages, and innate lymphoid cells, which are abundant in the intestine. At the intestinal mucosal surface, IL-23R gene expression and protein levels are found to be elevated in IBD patients. It is believed that IL-23 mediates this effect by promoting the development of pathogenic CD4+ T cell populations that produce IL-6, IL-17, and tumor necrosis factor (TNF).

[0003] Produced IL-23 is enriched in the intestine, which plays a key role in regulating the balance between tolerance and immunity through T cell-dependent and -independent pathways of intestinal inflammation by its effects on Th1 and Th17-related cytokines and by suppressing regulatory T cell responses in the intestine, which favor inflammation. In addition, polymorphisms of the IL-23 receptor (IL-23R) have been associated with susceptibility to inflammatory bowel disease (IBD), further establishing the key role of the IL-23 pathway in intestinal homeostasis.

[0004] Psoriasis is a chronic skin disease affecting 2% to 3% of the total population, which has been shown to be mediated by a mechanism of inflammatory T cell responses in the body. IL-23 is one of several interleukins that is believed to be a key player in the pathogenesis of psoriasis, which is said to maintain chronic autoimmune inflammation via induction of interleukin-17, regulation of memory T cells, and activation of macrophages. Expression of IL-23 and IL-23R has been shown to be increased in tissues of psoriasis patients, and antibodies that neutralize IL-23 show IL-23-dependent inhibition of psoriasis development in animal models of psoriasis.

[0005] IL-23 is a heterodimer composed of a unique pl9 subunit and the p40 subunit of IL-12, which is a helper T cell 1 (TH1) development cytokine involved in the production of interferon-gamma (IFN-γ). Although both IL-23 and IL-12 contain the p40 subunit, they have different phenotypic properties. For example, IL-12-deficient animals are susceptible to inflammatory autoimmune diseases, while IL-23-deficient animals are resistant, presumably because of a reduction in the number of IL-6-, IL-17-, and TNF-producing CD4+ T cells in the CNS of IL-23-deficient animals. IL-23 binds to IL-23R, which is a heterodimeric receptor composed of IL-12Rβ1 and IL-23R subunits. Binding of IL-23 to IL-23R activates Jak-stat signal transduction molecules, Jak2, Tyk2, and Stat1, Stat3, Stat4, and Stat5, although activation of Stat4 is substantially weaker, and a different DNA-binding Stat complex is formed in response to IL-23 compared to IL-12. IL-23R constitutively binds to Jak2 and binds to Stat3 in a ligand-dependent manner. In contrast to IL-12, which primarily acts on naive CD4(+) T cells, IL-23 preferentially acts on memory CD4(+) T cells.

[0006] Therapeutic moieties that inhibit the IL-23 pathway have been identified for the treatment of IL-23-associated diseases. A number of antibodies that bind to IL-23 or IL-23R have been identified, including ustekinumab (a humanized antibody that binds IL-23), which has been approved for the treatment of psoriasis. More recently, polypeptide inhibitors that bind to IL-23R and inhibit the binding of IL-23 to IL-23R have been identified. Clinical trials of ustekinumab and briakinumab, which target the common p40 subunit, and tildrakizumab, guselkumab, MEDI2070, and BI-655066, which target the unique pl9 subunit of IL-23, in Crohn's disease or psoriasis highlight the potential of blockade of IL-23 signaling in the treatment of human inflammatory diseases. While these findings are promising, there remains a challenge with respect to identifying stable and selective agents that preferentially target the IL-23 pathway in the gut, which can be used to treat intestinal inflammation, including Crohn's disease, ulcerative colitis, and related conditions.

[0007] Accordingly, there remains a need in the art for new therapies targeting the IL-23 pathway that can be used to treat and prevent IL-23 associated diseases, including autoimmune associated diseases. In addition, compounds and methods that specifically target IL-23R from the luminal side can provide therapeutic benefit to IBD patients. The present invention addresses these needs by providing novel peptide inhibitors that bind IL-23R to inhibit binding to and signaling by IL-23 and are suitable for oral use. SUMMARY

[0008] The present invention discloses a cyclic peptide inhibitor of interleukin-23 receptor, a stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, and use of the cyclic peptide inhibitor in treating or preventing diseases including inflammatory bowel disease, Crohn's disease, and psoriasis.

[0009] The cyclic peptide compound of the present invention has protein stability, is stable to plasma proteases, epithelial proteases, gastric and intestinal proteases, lung surface proteases, intracellular proteases, etc., has specific targeting to IL-23, has a long plasma half-life, and has good pharmacokinetic and pharmacodynamic properties.

[0010] The present invention relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the peptide compound has an amino acid sequence of Formula (I-A): Xa1-H-T-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-E-N-[3-Pal]-Xa 13 (I-A) (SEQ ID NO: 5)

[0011] The present invention also relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the peptide compound has an amino acid sequence of Formula (I), Formula (II): Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Glu-Asn-[3-Pal]-Xa 13 (I) (SEQ ID NO: 1), Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Xa 10 -Asn-[3-Pal]-Xa 13 (II) (SEQ ID NO: 2)

[0012] The present invention also relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the peptide compound has an amino acid sequence of Formula (IV): Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Glu-Asn-[3-Pal]-Xa 13(IV) (SEQ ID NO: 4)

[0013] In some embodiments, Xa1 is Pen or (D)Pen;

[0014] In some embodiments, Xa1 is Pen;

[0015] In some embodiments, Xa4 is Trp(7-methyl) or Trp(7-cyclopropyl);

[0016] In some embodiments, Xa5 is Lys or Gin;

[0017] In some embodiments, Xa7 is

[0018] In some embodiments, Xa7 is

[0019] In some embodiments, Xa9 is Thp or

[0020] In some embodiments, Xa 10 is Glu,

[0021] In some embodiments, Xa 13 is Sarc or

[0022] In some embodiments, Xa 13 is Sarc or is absent;

[0023] In some embodiments, the cyclic peptide compound is cyclized through a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0024] In some embodiments, at least one amino acid in the cyclic peptide compound is quaternized and / or conjugated with a modifying group, wherein the modifying group comprises a PEG linker or a quaternary ammonium;

[0025] In some embodiments, as an alternative, the cyclic peptide compound Xa1 is linked to the glutamic acid side chain in the sequence of Formula (I-A) through a modifying group to form a ring, wherein the modifying group comprises a PEG linker;

[0026] In some embodiments, provided that:

[0027] (1) when Xa4 is Trp(7-cyclopropyl), Xa5 is Gin, Xa9 is and only when a modifying group is conjugated at Xa1, the modifying group is not:

[0028] (2) when Xa4 is Trp (7-cyclopropyl), Xa5 is Lys, Xa9 is , and only when the modifying group is conjugated at Xa5, the modifying group is not:

[0029] In some embodiments, the cyclic peptide compound is conjugated at at least one of Xa1, Xa5, Xa7, E, Xa 13 , 3-Pal, or T to -L-R, L comprising a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -CO 1-6 haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid.

[0030] In some embodiments, the cyclic peptide compound is conjugated at at least one of Xa1, Xa5, Xa7, E, Xa 13 , 3-Pal, or T to -L-R, L comprising a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -CO 1-6 haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid.

[0031] In some embodiments, the cyclic peptide compound is conjugated at at least one of Xa1, Xa5, Xa7, E, or Sarc to -L-R, L comprising a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -CO 1-6 haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid.

[0032] In some embodiments, the cyclic peptide compound is conjugated at at least one of Xa1, Xa5, or Xa7 to -L-R, L comprising a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -CO 1-6 haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid.

[0033] In some embodiments, the cyclic peptide compound is conjugated at at least one of Xa1, Xa5, Xa7, E, Xa 13At least one of the following conjugated modifying groups: 3-Pal or T, wherein the modifying group is -LR, L is -L1-L2-L3-, wherein L3 is connected to R, and L1 is a bond, L2 contains a PEG linker, L3 is a spacer unit, and R is: n is 4-15;

[0034] In some embodiments, the cyclic peptide compound Xa1 forms a ring with the glutamate side chain in the sequence of formula (I) via a -L-link;

[0035] In some embodiments, the PEG connector is p is 0-50, q is 1-50;

[0036] In some embodiments, the PEG connector is p is 0-50, q is 1-50;

[0037] In some embodiments, R is hydrogen, hydroxyl, C 1-3 Alkyl, -COC 1-3 Alkyl, -CO halogenated C 1-3 Alkyl, C 1-24 Fatty acids or C 1-24 heterofatty acids;

[0038] In some implementations, the C 1-30 Fatty acids or C 1-30 The following are heterofatty acids: Where n is 1-15;

[0039] In some implementations, the C 1-20 Fatty acids or C 1-20 The following are heterofatty acids: Where n is 1-15;

[0040] In some implementations, the C 1-24 Fatty acids or C 1-24 The following are heterofatty acids: Where n is 2-15;

[0041] In some implementations, the C 1-24 Fatty acids or C 1-24 The following are heterofatty acids: Where n is 4-12;

[0042] In some implementations, the C 1-30 Fatty acids or C 1-30 The following are heterofatty acids:

[0043] In some embodiments, the C 1-24 fatty acid or C 1-24 heterofatty acid is:

[0044] In some embodiments, the L is -L1-L2-L3-, wherein L2 is p is 0-50, q is 1-50; each of L1and L3is independently a bond, a combination of one or more of the foregoing, m is 0-5, in some embodiments, m is 1-5;

[0045] In some embodiments, L2is p is 0-50, q is 1-50, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is

[0046] In some embodiments, the L2is p is 0-50, q is 1-50;

[0047] In some embodiments, p is 1-3, q is 1-2;

[0048] In some embodiments, the L2is p is 0-10, q is 1-5;

[0049] In some embodiments, the L2is p is 1-6, q is 1-4;

[0050] In some embodiments, the L2is p is 1-3, q is 1-2;

[0051] In some embodiments, the L2is p is 1-3, q is 1-2;

[0052] In some embodiments, the L2is

[0053] In some embodiments, each of L1and L3is independently a bond, a combination of one or more of the following:

[0054] In some embodiments, the L1is a bond, a combination of one or more of the following:

[0055] In some embodiments, the L3is a combination of one or more of the following:

[0056] In some embodiments, the L3is In some embodiments, the L3is

[0057] In some embodiments, the L is: p is 1-3, q is 1-2, m is 1-2;

[0058] In some embodiments, the L is: p is 1-3, q is 1-2;

[0059] In some embodiments, the L is:

[0060] In some embodiments, the L is:

[0061] In some embodiments, the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E, Xa 13 3-Pal, or T;

[0062] In some embodiments, the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E, Xa 13 3-Pal, or T;

[0063] In some embodiments, the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E, Xa

[0064] In some embodiments, the cyclic peptide compound is quaternized and / or conjugated with a modifying group comprising a quaternary ammonium at at least one of Xa1, Xa5, or Xa7.

[0065] The present application also relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence according to Formula (II-1), (II-2), (II-3), (II-4):

[0066] In some embodiments, the two cyclic peptide compounds are linked to form a peptide dimer via a polyethylene glycol;

[0067] In some embodiments, the two cyclic peptide compounds are linked to form a peptide dimer via In some embodiments, the two cyclic peptide compounds are linked to form a peptide dimer via

[0068] In some embodiments, the two cyclic peptide compounds are linked to form a peptide dimer via In some embodiments, the two cyclic peptide compounds are linked to form a peptide dimer via

[0069] The present application also relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence according to Formula (II-1), (II-2), (II-3), (II-4):

[0070] Pen-H-T-[Trp(7-methyl)]-Gln-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-1) (SEQ ID NO: 6)

[0071] Pen-H-T-[Trp(7-methyl)]-Lys-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-2) (SEQ ID NO: 7)

[0072] Xa1-H-T-[Trp(7-methyl)]-Lys-Xa6-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-3) (SEQ ID NO: 8)

[0073] Xa1-H-T-[Trp(7-methyl)]-Lys-Xa6-Xa7-[2-Nal]-Xa9-E-N-[3-Pal]-[Sarc] (II-4) (SEQ ID NO: 9)

[0074] Xa1, Xa6are Pen or and Xa1, Xa6are not both Pen;

[0075] Xa7is

[0076] Xa9is

[0077] the cyclic peptide compound is cyclized by a Pen-Pen or Xa1-Xa6disulfide bond;

[0078] alternatively, the side chain terminal amino group of amino acid Pen, Gln, Lys, or Xa7or Xa1is condensed with an α-hydroxyacetic acid, β-hydroxypropionic acid, 2-methyl lactic acid, an amino acid, or an α-hydroxyisobutyric acid;

[0079] the cyclic peptide compound is optionally linked to a protecting group;

[0080] with the proviso that the cyclic peptide compound is not

[0081] In some embodiments, the protecting group is a hydroxyl, NH2, C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -COC 1-6 alkyl, -COhaloC 1-6 alkyl, Gly, Ser;

[0082] In some embodiments, the protecting group is a hydroxyl, NH2, C 1-3 alkyl, -NHC 1-3 alkyl, -N(C 1-3 alkyl)2, -COC 1-3 alkyl, or -COhaloC 1-3 alkyl;

[0083] In some embodiments, the protecting group is a hydroxyl, NH2, methyl, -N(CH3)2, -COCH3, or -COCF3;

[0084] In some embodiments, the protecting group is an amino protecting group selected from acetyl;

[0085] In some embodiments, the side chain terminal amino group of amino acid Pen, Gln, Lys, or Xa7or Xa1is condensed with an α-hydroxyacetic acid, β-hydroxypropionic acid, serine, or an α-hydroxyisobutyric acid;

[0086] In some embodiments, the terminal amino group of the side chain of amino acids Pen, Gln, Lys, or Xa7 is condensed and linked with α-hydroxyacetic acid, β-hydroxypropionic acid, or serine.

[0087] In some implementations, n is 4, 5, 6, 7, 8, 9, 10, 11, or 12;

[0088] In some implementations, p is 1, 2, 3, 4, 5, or 6;

[0089] In some implementations, q is 1, 2, 3, or 4;

[0090] In some implementations, m is 0, 1, 2, 3, 4, or 5;

[0091] In some implementations, m is 0, 1, or 2;

[0092] In some implementations, m is 1, 2, 3, 4, or 5;

[0093] In some implementations, t is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10.

[0094] Specifically, the first embodiment relates to a cyclic peptide compound, its stereoisomer or its pharmaceutically acceptable salt, wherein the cyclic peptide compound has an amino acid sequence of formula (IA):

[0095] Xa1-HT-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-EN-[3-Pal]-Xa 13 (IA)(SEQ ID NO: 5)

[0096] in:

[0097] Xa1 is Pen;

[0098] Xa4 is Trp(7-methyl) or Trp(7-cyclopropyl);

[0099] Xa5 is either Gln or Lys;

[0100] Xa7 is

[0101] Xa9 is Thrp or

[0102] Xa 13 It is either Sarc or does not exist;

[0103] The cyclic peptide compound is cyclized via a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0104] at least one amino acid in the cyclic peptide compound is quaternized and / or conjugated with a modifying group, wherein the modifying group comprises a PEG linker or a quaternary ammonium;

[0105] Alternatively, the cyclic peptide compound Xa1is linked to the glutamic acid side chain in the sequence of Formula (I-A) through a modifying group, wherein the modifying group comprises a PEG linker;

[0106] provided that:

[0107] (1) when Xa4is Trp(7-cyclopropyl), Xa5is Gln, Xa9is and only when a modifying group is conjugated at Xa1, the modifying group is not:

[0108] (2) when Xa4is Trp(7-cyclopropyl), Xa5is Lys, Xa9is and only when a modifying group is conjugated at Xa5, the modifying group is not:

[0109] In a particular second embodiment, there is provided a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of Formula (I-A):

[0110] Xa1-H-T-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-E-N-[3-Pal]-[Sarc] (I-A) (SEQ ID NO: 5)

[0111] wherein:

[0112] Xa1is Pen;

[0113] Xa4is Trp(7-methyl) or Trp(7-cyclopropyl);

[0114] Xa5is Gln or Lys;

[0115] Xa7is

[0116] Xa9is Thp or

[0117] the cyclic peptide compound is cyclized through a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0118] at least one amino acid in the cyclic peptide compound is quaternized and / or conjugated with a modifying group, wherein the modifying group comprises a PEG linker or a quaternary ammonium;

[0119] alternatively, the cyclic peptide compound Xa1 is linked with the glutamic acid side chain in the sequence of Formula (I-A) to form a ring with a modifying group, wherein the modifying group comprises a PEG linker;

[0120] provided that:

[0121] (1) when Xa4 is Trp (7-cyclopropyl), Xa5 is Gin, Xa9 is and only when a modifying group is conjugated at Xa1, the modifying group is not:

[0122] (2) when Xa4 is Trp (7-cyclopropyl), Xa5 is Lys, Xa9 is and only when a modifying group is conjugated at Xa5, the modifying group is not:

[0123] a particular third embodiment, wherein the cyclic peptide compound is conjugated with -L-R at at least one of Xa1, Xa5, Xa7, E, Xa 13 , 3-Pal, or T, L comprises a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -COC 1-6 alkyl, -CO haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid; alternatively, the cyclic peptide compound Xa1 is linked with the glutamic acid side chain in the sequence of Formula (I-A) to form a ring with -L-.

[0124] a particular fourth embodiment, wherein the cyclic peptide compound is conjugated with -L-R at at least one of Xa1, Xa5, Xa7, E, Xa 13 , or 3-Pal, L comprises a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -COC 1-6 alkyl, -CO haloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid; alternatively, the cyclic peptide compound Xa1 is linked with the glutamic acid side chain in the sequence of Formula (I-A) to form a ring with -L-.

[0125] In a particular fifth embodiment, wherein the cyclic peptide compound is conjugated at at least one of Xal, Xa5, Xa7, E, or Sarc with -L-R, L comprises a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -COhaloC 1-6 alkyl, -COhaloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid; alternatively, the cyclic peptide compound Xal forms a ring with the glutamic acid side chain in the sequence of Formula (I-A) through a -L- linkage.

[0126] In a particular sixth embodiment, wherein the cyclic peptide compound is conjugated at at least one of Xal, Xa5, or Xa7 with -L-R, L comprises a PEG linker, and R is hydrogen, hydroxyl, C 1-6 alkyl, -COhaloC 1-6 alkyl, -COhaloC 1-6 alkyl, C 1-30 fatty acid, or C 1-30 heterofatty acid; alternatively, the cyclic peptide compound Xal forms a ring with the glutamic acid side chain in the sequence of Formula (I-A) through a -L- linkage.

[0127] In a particular seventh embodiment, wherein the C 1-30 fatty acid, or C 1-30 heterofatty acid is: wherein n is 1-15.

[0128] In a particular eighth embodiment, wherein the C 1-24 fatty acid, or C 1-24 heterofatty acid is: wherein n is 2-15.

[0129] In a particular ninth embodiment, wherein the L is -L1-L2-L3-, wherein L2 is p is 0-50, q is 1-50; each of L1and L3is independently a bond, a combination of one or more of the foregoing, and m is 0-5;

[0130] Preferably, L is: p is 1-3, q is 1-2, and m is 1-2.

[0131] In a particular tenth embodiment, wherein the L is -L1-L2-L3-, wherein L2 is p is 0-50, q is 1-50; each of L1and L3is independently a bond, one or more combinations of the following: m is 0-5;

[0132] In some embodiments, the L is: p is 1-3, q is 1-2.

[0133] Specific eleventh embodiment, wherein the L is -L1-L2-L3-, wherein L2is p is 0-50, q is 1-50; each of L1and L3is independently a bond, one or more combinations of the following: m is 0-5;

[0134] In some embodiments, the L is: p is 1-3, q is 1-2.

[0135] Specific twelfth embodiment, wherein the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E, Xa 13 3-Pal or T.

[0136] Specific thirteenth embodiment, wherein the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E, Xa 13 or 3-Pal.

[0137] Specific fourteenth embodiment, wherein the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5, Xa7, E or Sarc.

[0138] Specific fifteenth embodiment, wherein the cyclic peptide compound is quaternized and / or conjugated with a modification group comprising a quaternary ammonium at at least one of Xa1, Xa5 or Xa7.

[0139] In a sixteenth embodiment, a peptide dimer compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof is provided, wherein two cyclic peptide compounds having an amino acid sequence of Formula (I) as described in any of the preceding embodiments are linked via one or more linkers selected from diethylene glycol, iminodiacetic acid (IDA), β-Ala-iminodiacetic acid (β-Ala-IDA), or polyethylene glycol to form a peptide dimer.

[0140] In a seventeenth embodiment, a cyclic peptide compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof is provided, wherein the cyclic peptide compound has an amino acid sequence of Formula (II-1), (II-2), (II-3), (II-4):

[0141] Pen-H-T-[Trp(7-methyl)]-Gln-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-1) (SEQ ID NO: 6)

[0142] Pen-H-T-[Trp(7-methyl)]-Lys-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-2) (SEQ ID NO: 7)

[0143] Xa1-H-T-[Trp(7-methyl)]-Lys-Xa6-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-3) (SEQ ID NO: 8)

[0144] Xa1-H-T-[Trp(7-methyl)]-Lys-Xa6-Xa7-[2-Nal]-Xa9-E-N-[3-Pal]-[Sarc] (II-4) (SEQ ID NO: 9)

[0145] Xa1, Xa6 is Pen or and Xa1, Xa6 are not both Pen;

[0146] Xa7 is

[0147] Xa9 is

[0148] the cyclic peptide compound is cyclized by a Pen-Pen or Xa1-Xa6 disulfide bond;

[0149] alternatively, the side chain terminal amino group of amino acid Pen, Gin, Lys, Xa7, or Xa1 is condensed with an a-hydroxyacetic acid, a β-hydroxypropionic acid, 2-methyl lactic acid, an amino acid, an a-hydroxyisobutyric acid;

[0150] the cyclic peptide compound is optionally linked to a protecting group;

[0151] provided that the cyclic peptide compound is not

[0152] In a particular eighteenth embodiment, there is provided a cyclic peptide compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of formula (II-1), (II-2):

[0153] Pen-H-T-[Trp(7-methyl)]-Gln-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-1) (SEQ ID NO: 6)

[0154] Pen-H-T-[Trp(7-methyl)]-Gln-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-1) (SEQ ID NO: 6)

[0155] Xa7 is

[0156] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0157] alternatively, the side chain terminal amino group of amino acid Pen, Gin, Lys, Xa7, or Xa1 is condensed with an a-hydroxyacetic acid, a β-hydroxypropionic acid, 2-methyl lactic acid, an amino acid, an a-hydroxyisobutyric acid;

[0158] provided that the cyclic peptide compound is not

[0159] In a particular nineteenth embodiment, there is provided a cyclic peptide compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of formula (II-1), (II-2):

[0160] Pen-H-T-[Trp(7-methyl)]-Gln-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-1) (SEQ ID NO: 6)

[0161] Pen-H-T-[Trp(7-methyl)]-Lys-[Pen]-Xa7-[2-Nal]-[Thp]-E-N-[3-Pal]-[Sarc] (II-2) (SEQ ID NO: 7)

[0162] Xa7is

[0163] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0164] alternatively, the side chain terminal amino group of the amino acids Pen, Gln, Lys is condensed with an alpha-hydroxyacetic acid, a beta-hydroxypropionic acid, an amino acid;

[0165] with the proviso that the cyclic peptide compound is not

[0166] In a particular twentieth embodiment, there is provided a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of formula (I), (II):

[0167] Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Glu-Asn-[3-Pal]-Xa 13 (I) (SEQ ID NO: 1)

[0168] Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Xa 10 -Asn-[3-Pal]-Xa 13 (II) (SEQ ID NO: 2)

[0169] wherein:

[0170] Xa1is Pen or (D)Pen;

[0171] Xa4is Trp(7-methyl) or Trp(7-cyclopropyl);

[0172] Xa5is Gln or Lys;

[0173] Xa7is

[0174] Xa9is Thp or

[0175] Xa 10 is Glu,

[0176] Xa 13 is Sarc or

[0177] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group; in some embodiments, the protecting group is hydroxyl, NH2, C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -COC 1-6 alkyl, -COhaloC 1-6 alkyl, Gly, or Ser; in some embodiments, the protecting group is hydroxyl, NH2, C 1-3 alkyl, -NHC 1-3 alkyl, -N(C 1-3 alkyl)2, -COC 1-3 alkyl, or -COhaloC 1-3 alkyl; in some embodiments, the protecting group is hydroxyl, NH2, methyl, -N(CH3)2, -COCH3, or -COCF3;

[0178] the cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, Glu, Xa 10 , Xa 13 , 3-Pal, or Thr; in some embodiments, the cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, Glu, Xa 13 , 3-Pal, or Thr; the modifying group is -L-R, L is -L1-L2-L3-, wherein L3 is attached to R, and L1 is a bond, L2 comprises a PEG linker, L3 is a spacer unit, and R is: n is 4-15;

[0179] provided that:

[0180] (1) when Xa4 is Trp(7-cyclopropyl), Xa5 is Gln, Xa9 is and the modifying group is not:

[0181] (2) when Xa4 is Trp(7-cyclopropyl), Xa5 is Lys, Xa9 is and the modifying group is not:

[0182] A twenty-first embodiment relates to a cyclic peptide compound, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of Formula (I):

[0183] Xa1-H-T-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-E-N-[3-Pal]-Xa 13 (I) (SEQ ID NO: 1)

[0184] wherein:

[0185] Xa1is Pen or (D)Pen; in some embodiments, Xa1is Pen;

[0186] Xa4is Trp(7-methyl) or Trp(7-cyclopropyl);

[0187] Xa5is Gin or Lys;

[0188] Xa7is in some embodiments, Xa7is

[0189] Xa9is Thp or

[0190] Xa 13 is Sarc or in some embodiments, Xa 13 is Sarc or

[0191] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0192] the cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, E, Xa 13 , 3-Pal, or T, the modifying group being -L-R, L being -L1-L2-L3-, wherein L3is attached to R, and L1is a bond, L2comprises a PEG linker, L3is a spacer unit, and R is: n is 4-15; in some embodiments, the cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, E, Xa 13 , 3-Pal, or T, the modifying group being -L-R, L being -L1-L2-L3-, wherein L3is attached to R, and L1is a bond, L2is a PEG linker, L3is a spacer unit, and R is: n is 4-15;

[0193] provided that:

[0194] (1) when Xa4is Trp(7-cyclopropyl), Xa5is Gln, Xa9is and only when the modifying group is conjugated at Xa1, the modifying group is not:

[0195] (2) when Xa4is Trp(7-cyclopropyl), Xa5is Lys, Xa9is and only when the modifying group is conjugated at Xa5, the modifying group is not:

[0196] In a twenty-second embodiment, the application relates to a cyclic peptide compound as described in the twentieth, twenty-first embodiments, stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein L is -L1-L2-L3- wherein:

[0197] L1is a bond;

[0198] L2is p is 0-50, q is 1-50, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is In some embodiments, L2is p is 0-50, q is 1-50, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is In some embodiments, L2is p is 0-50, q is 1-50; in some embodiments, p is 1-3, q is 1-2;

[0199] L3is a bond, a combination of one or more of a combination of one or more of In some embodiments, L3is In some embodiments, L3is

[0200] In a twenty-third embodiment, the application relates to a cyclic peptide compound as described in any one of the twentieth, twenty-first, or twenty-second embodiments, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0201] Xa1is Pen;

[0202] Xa4is Trp(7-methyl);

[0203] Xa5is Lys;

[0204] Xa7is

[0205] Xa9is Thp.

[0206] In a twenty-fourth embodiment, the application relates to a cyclic peptide compound as described in any one of the twentieth, twenty-first, twenty-second, or twenty-third embodiments, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein:

[0207] Xa1is Pen;

[0208] Xa4is Trp(7-methyl);

[0209] Xa5is Lys;

[0210] Xa7is

[0211] Xa9is Thp;

[0212] Xa 10 is Glu;

[0213] Xa 13 is Sarc;

[0214] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group;

[0215] the cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, Glu, Xa 13 , 3-Pal, or Thr.

[0216] In a twenty-fifth embodiment, the application relates to a cyclic peptide compound as described in any one of the twentieth, twenty-first, twenty-second, twenty-third, or twenty-fourth embodiments, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein the L is: p is 1-5, preferably p is 1 or 2, q is 1-2, and X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is

[0217] In some embodiments, L is: p is 1-5, preferably p is 1 or 2, q is 1-2, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is In some embodiments, L is p is 1-3, q is 1-2; in some embodiments, L is: p is 1-3, q is 1-2;

[0218] In some embodiments, L is: X is

[0219] In some embodiments, L is: p is 1-5, q is 1-2, X is preferably L is: p is 1-5, q is 1-2, X is more preferably L is: X is

[0220] In a twenty-sixth embodiment, the application relates to a cyclic peptide compound according to any one of the twentieth, twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiments, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R is: n is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15; in some embodiments, R is:

[0221] In a twenty-seventh embodiment, the application relates to a cyclic peptide compound according to any one of the twentieth, twenty-first, twenty-second, twenty-third, twenty-fourth, twenty-fifth, or twenty-sixth embodiments, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein Xa1, Xa5, Xa 13the amino acid residues at Xa1and Xa5are optionally linked to a protecting group; in some embodiments, the protecting group of the amino acid residues at Xa1and Xa5is selected from the group consisting of acetyl, and / or, the protecting group of the amino acid residue at Xa 13 the protecting group of the amino acid residue at Xa

[0222] In a twenty-eighth embodiment, the application relates to a cyclic peptide compound as described in any one of the twenty-first, twenty-second, twenty-third, twenty-fourth, twenty-fifth, twenty-sixth, or twenty-seventh embodiments, wherein the cyclic peptide compound is conjugated to a modifying group at Glu.

[0223] In a twenty-ninth embodiment, the application relates to a cyclic peptide compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of Formula (III),

[0224] [Pen]-His-Thr-Trp(7-Me)-Lys(Ac)-[Pen]-Xa7-[2-Nal]-THP-Xa 10 -Asn-[3-Pal]-Sarc(III) (SEQ ID NO: 3)

[0225] Xa7is

[0226] Xa 10 is

[0227] the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group; in some embodiments, the protecting group is hydroxyl, NH2, C 1-6 alkyl, -NHC 1-6 alkyl, -N(C 1-6 alkyl)2, -COC 1-6 alkyl, -COhaloC 1-6 alkyl, Gly, or Ser; in some embodiments, the protecting group is hydroxyl, NH2, C 1-3 alkyl, -NHC 1-3 alkyl, -N(C 1-3 alkyl)2, -COC 1-3 alkyl, or -COhaloC 1-3 alkyl; in some embodiments, the protecting group is hydroxyl, NH2, methyl, -N(CH3)2, -COCH3, or -COCF3.

[0228] Specifically, in the thirtieth embodiment, a peptide dimer compound, its stereoisomer or its pharmaceutically acceptable salt, wherein two cyclic peptide compounds are linked via one or more linkers selected from diethylene glycol, iminodiacetic acid, β-Ala-iminodiacetic acid, or polyethylene glycol to form a peptide dimer, said cyclic peptide compounds having the amino acid sequences of formulas (I) and (II) as described in the twentieth embodiment. Further, in some embodiments, a peptide dimer compound as described in the twenty-eighth embodiment is involved, wherein the two cyclic peptide compounds are linked via polyethylene glycol to form a peptide dimer; in some embodiments, the two cyclic peptide compounds are linked via... The two cyclic peptide compounds are linked to form a peptide dimer, with a t value of 0-99; in some embodiments, the two cyclic peptide compounds are... They connect to form peptide dimers, with t ranging from 0 to 10.

[0229] Specifically, the thirty-first embodiment involves a compound as described in any of the foregoing technical solutions, the stereoisomer of which or its pharmaceutically acceptable salt is selected from one of the structures in Table 1 below:

[0230] Table 1:

[0231] The present invention also relates to a pharmaceutical composition comprising the peptide compound or a pharmaceutically acceptable salt thereof described in any of the foregoing technical solutions, and a pharmaceutically acceptable carrier and / or excipient.

[0232] The present invention also relates to an application of the peptide compound or its pharmaceutically acceptable salt, or the pharmaceutical composition described in any of the foregoing technical solutions, in the preparation of a medicament for the prevention and treatment of diseases or conditions in which IL-23 is overexpressed in diseased tissues of a subject.

[0233] Furthermore, the diseases or conditions that overexpress IL-23 include inflammatory bowel disease, Crohn's disease, and psoriasis.

[0234] The present invention also relates to a pharmaceutical composition or pharmaceutical formulation comprising 1-1500 mg of the peptide compound or a pharmaceutically acceptable salt thereof as described in any of the foregoing technical solutions, and a pharmaceutically acceptable carrier and / or excipient.

[0235] The present application also relates to a method for treating a disease in a mammal or a human, the method comprising administering to the subject a therapeutically effective amount of the peptide compound or a pharmaceutically acceptable salt thereof according to any one of the preceding embodiments, preferably 1-1500 mg, preferably the disease is inflammatory bowel disease, Crohn's disease and psoriasis.

[0236] The present application also provides a composition or a pharmaceutical preparation comprising the peptide compound or a pharmaceutically acceptable salt thereof according to any one of the preceding embodiments, and a pharmaceutically acceptable carrier and / or adjuvant. The pharmaceutical composition can be in the form of a unit preparation (unit preparation is also referred to as "formulation specification").

[0237] Further, the present application provides a composition or a pharmaceutical preparation comprising 1-1500 mg of the peptide compound or a pharmaceutically acceptable salt thereof according to any one of the preceding embodiments, and a pharmaceutically acceptable carrier and / or adjuvant.

[0238] The present application also provides the use of the peptide compound or a pharmaceutically acceptable salt thereof according to any one of the preceding embodiments in the preparation of a medicament for preventing and treating a disease or a condition in which IL-23 is overexpressed in a subject. Further, the disease or the condition in which IL-23 is overexpressed includes inflammatory bowel disease, Crohn's disease and psoriasis.

[0239] The present application also provides a method for treating a disease in a mammal or a human, the method comprising administering to the subject a therapeutically effective amount of the peptide compound or a pharmaceutically acceptable salt thereof according to any one of the preceding embodiments, preferably 1-1500 mg, preferably the disease is inflammatory bowel disease, Crohn's disease and psoriasis. In some embodiments, the mammal in the present application does not include a human.

[0240] An "effective amount" or "therapeutically effective amount" as used herein refers to an amount of a compound disclosed herein that, when administered to a subject for treating a disease or condition, will relieve to some extent one or more of the symptoms of the disease or condition being treated. In some embodiments, the result is a decrease and / or alleviation of signs, symptoms, or causes of a disease, or any other desired alteration of a biological system. For example, an "effective amount" for therapeutic uses is the amount of a compound disclosed herein, or a pharmaceutically acceptable salt thereof, that will elicit the biological or medical response (e.g., reduce or alleviate disease symptoms) in a subject. Examples of therapeutic effects include, but are not limited to, 1-1500 mg, 1-1400 mg, 1-1300 mg, 1-1200 mg, 1-1000 mg, 1-900 mg, 1-800 mg, 1-700 mg, 1-600 mg, 1-500 mg, 1-400 mg, 1-300 mg, 1-250 mg, 1-200 mg, 1-150 mg, 1-125 mg, 1-100 mg, 1-80 mg, 1-60 mg, 1-50 mg, 1-40 mg, 1-25 mg, 1-20 mg, 5-1500 mg, 5-1000 mg, 5-900 mg, 5-800 mg, 5-700 mg, 5-600 mg, 5-500 mg, 5-400 mg, 5-300 mg, 5-250 mg, 5-200 mg, 5-150 mg, 5-125 mg, 5-100 mg, 5-90 mg, 5-70 mg, 5-80 mg, 5-60 mg, 5-50 mg, 5-40 mg, 5-30 mg, 5-25 mg, 5-20 mg, 10-1500 mg, 10-1000 mg, 10-900 mg, 10-800 mg, 10-700 mg, 10-600 mg, 10-500 mg, 10-450 mg, 10-400 mg, 10-300 mg, 10-250 mg, 10-200 mg, 10-150 mg, 10-125 mg, 10-100 mg, 10-90 mg, 10-80 mg, 10-70 mg, 10-60 mg, 10-50 mg, 10-40 mg, 10-30 mg, 10-20 mg; 20-1500 mg, 20-1000 mg, 20-900 mg, 20-800 mg, 20-700 mg, 20-600 mg, 20-500 mg, 20-400 mg, 20-350 mg, 20-300 mg, 20-250 mg, 20-200 mg, 20-150 mg, 20-125 mg, 20-100 mg, 20-90 mg, 20-80 mg, 20-70 mg, 20-60 mg, 20-50 mg, 20-40 mg, 20-30 mg;50-1500 mg, 50-1000 mg, 50-900 mg, 50-800 mg, 50-700 mg, 50-600 mg, 50-500 mg, 50-400 mg, 50-300 mg, 50-250 mg, 50-200 mg, 50-150 mg, 50-125 mg, 50-100 mg; 100-1500 mg, 100-1000 mg, 100-900 mg, 100-800 mg, 100-700 mg, 100-600 mg, 100-500 mg, 100-400 mg, 100-300 mg, 100-250 mg, 100-200 mg;

[0241] In some embodiments, the pharmaceutical composition or formulation of the present application contains a therapeutically effective amount of the peptide compound of the present application or a pharmaceutically acceptable salt thereof as described above;

[0242] The present application relates to a pharmaceutical composition or a pharmaceutical formulation comprising a therapeutically effective amount of the peptide compound of the present application or a pharmaceutically acceptable salt thereof and a carrier and / or an excipient. The pharmaceutical composition can be in the form of a unit formulation (the amount of the main drug in the unit formulation is also referred to as "formulation specification"). In some embodiments, the pharmaceutical composition includes, but is not limited to, 1 mg, 1.25 mg, 2.5 mg, 5 mg, 10 mg, 12.5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 275 mg, 300 mg, 325 mg, 350 mg, 375 mg, 400 mg, 425 mg, 450 mg, 475 mg, 500 mg, 525 mg, 550 mg, 575 mg, 600 mg, 625 mg, 650 mg, 675 mg, 700 mg, 725 mg, 750 mg, 775 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg of the peptide compound of the present application or a pharmaceutically acceptable salt thereof.

[0243] A method for treating a disease in a mammal or human, the method comprising administering to the subject a therapeutically effective amount of a peptide compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient, the therapeutically effective amount preferably being 1-1500 mg, the disease preferably being inflammatory bowel disease, Crohn's disease, and psoriasis.

[0244] A method for treating a disease in a mammal or human, the method comprising administering to the subject a therapeutically effective amount of a peptide compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient, the therapeutically effective amount preferably being 1-1500 mg, the disease preferably being inflammatory bowel disease, Crohn's disease, and psoriasis.

[0245] The present invention relates to a kit which can comprise a composition in single or multiple dose form, the kit comprising a peptide compound of the present invention or a pharmaceutically acceptable salt thereof, the amount of the compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt thereof being the same as the amount thereof in the pharmaceutical composition described above.

[0246] The amount of the compound of the present invention or a stereoisomer or a pharmaceutically acceptable salt thereof in the present invention is in each case calculated as the free base.

[0247] "Formulation strength" means the weight of the principal drug contained in each unit of a preparation.

[0248] The term

[0249] Unless otherwise defined, scientific and technical terms used in this application shall have the meanings that are commonly understood by one of ordinary skill in the art. Generally, nomenclature used in connection with, and techniques of, chemistry, molecular biology, cell and cancer biology, immunology, microbiology, pharmacology, and protein and nucleic acid chemistry described herein are those well-known and commonly used in the art. In the event that there is a contradiction between a definition provided herein and that generally understood by one of ordinary skill in the art, the definition provided herein controls. Where the name of a commodity appears herein, it is intended to refer to its corresponding commodity or active ingredient thereof. All patents, published patent applications and publications cited herein are hereby incorporated by reference.

[0250] As used herein, the following terms have the meanings ascribed to them unless otherwise specified.

[0251] Throughout this specification, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.

[0252] "Peptide" refers broadly to a sequence of two or more amino acids joined together by peptide bonds. It is to be understood that this term does not imply a particular length of the amino acid polymer nor is it intended to suggest or distinguish whether the polypeptide was produced using recombinant techniques, chemical synthesis or enzymatic synthesis or whether it is naturally occurring.

[0253] The peptides provided herein can comprise at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity. Thus, the polypeptides provided herein comprise at least 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the sequence of Formula (I).

[0254] “Sequence identity,”“percentage of identity,”“percentage of homology,” or the like, including for example a“sequence 50% identical to,” refers to the extent to which sequences are identical within a comparison window of nucleotides or amino acids. Thus, a“percentage of sequence identity” can be calculated by comparing two optimally aligned sequences over the window, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, I) or identical amino acid residue (e.g., Ala, Pro, Ser, Thr, Gly, Val, Leu, He, Phe, Tyr, Trp, Lys, Arg, His, Asp, Glu, Asn, Gin, Cys, and Met) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the comparison window (i.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. Sequence similarity or sequence identity between sequences (which terms are used interchangeably herein) can be calculated as follows. To determine the percent identity of two amino acid sequences or two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In certain embodiments, the length of a reference sequence aligned for comparison purposes is at least 30%, preferably at least 40%>, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position.

[0255] The percentage of identity between two sequences is the number of positions at which the sequences share identity (taking into account the number of gaps that must be introduced for optimal alignment of the two sequences and the length of each gap) over the total number of positions in the sequences.

[0256] It is understood that the peptide sequences disclosed herein are shown from left to right, with the left end of the sequence being the N-terminus of the peptide and the right end of the sequence being the C-terminus of the peptide. The sequences disclosed herein are sequences incorporating a "Hy-" moiety at the amino terminal (N-terminal) end of the sequence and a "-OH" moiety or a "-NH2" moiety at the carboxy terminal (C-terminal) end of the sequence. In such cases, and unless otherwise indicated, the "Hy-" moiety at the N-terminal end of the sequence under discussion represents a hydrogen atom, corresponding to the presence of a primary or secondary amino group free at the N-terminal end, while the "-OH" or "NH2" moiety at the C-terminal end of the sequence represents a hydroxyl or amino group, respectively, corresponding to the presence of an amide group (CONH2) at the C-terminal end. In the sequences of the application, the C-terminal "-OH" moiety can be replaced by a C-terminal "-NH2" moiety, and vice versa.

[0257] The term "protecting group" refers to protecting groups at the free amino and / or carboxyl terminal end of the amino acid residues of the polypeptide, the protecting group preferably including, but not limited to, acetyl, amide, alkyl of 3-20 carbon atoms, Fmoc, t-boc, 9-fluoreneacetyl, 1-fluorene carboxy, 9-fluorene carboxy, 9-fluorenone-1-carboxy, benzyloxycarbonyl, xanthenyl (Xan), trityl (Trt), 4-methyltrityl (Mtt), 4-methoxytrityl (Mmt), 4-methoxy-2,3,6-trimethyl-benzenesulfonyl (Mtr), 1,3,5-trimethylbenzene (Mesitylene)-2-sulfonyl (Mts), 4,4-dimethoxybenzhydryl (Mbh), tosyl (Tos), 2,2,5,7,8-pentamethylchroman-6-sulfonyl (Pmc), 4-methylbenzyl (MeBzl), 4-methoxybenzyl (MeOBzl), benzyloxy (BzIO), benzyl (Bzl), benzoyl (Bz), 3-nitro-2-pyridinesulphenyl (Npys), 1-(4,4-dimentyl-2,6-diazenocyclohexyl)ethyl (Dde), 2,6-dichlorobenzyl (2,6-DiCl-Bzl), 2-chlorobenzyloxycarbonyl (2-Cl-Z), 2-bromobenzyloxycarbonyl (2-Br-Z), benzyloxymethyl (Bom), t-butyloxycarbonyl (Boc), cyclohexyloxy (cHxO), t-butyloxymethyl (Bum), t-butyloxy (tBuO), t-butyl (tBu), acetyl (Ac), benzoyl, benzyloxycarbonyl, propyl, butyl, pentyl, hexyl, and trifluoroacetyl (TFA). In some embodiments, the polypeptides of the application contain a protecting group coupled to the amino terminal end of the polypeptide, the amino terminal protecting group being a C 1-6 alkyl, -COC 1-6 alkyl or -COhaloC 1-6alkyl. In some embodiments, the polypeptides of the present application contain a protecting group coupled to the carboxyl terminus, the carboxyl terminal protecting group is NH2, -NHC 1-3 alkyl, -N(C 1-3 alkyl)2.

[0258] The term "NH2" can refer to a free amino group present at the amino terminus of a polypeptide. The term "OH" as used herein can refer to a free carboxyl group present at the carboxyl terminus of a peptide. Additionally, the term "Ac" as used herein refers to an acetyl protecting group formed by acylation of the C-terminus or N-terminus of a polypeptide. In certain peptides shown herein, NH2at the C-terminus of a peptide indicates an amino group.

[0259] The term "quaternary ammonium" takes its ordinary meaning in the art. For example, a quaternary ammonium is a substituent that contains one or more permanently positively charged nitrogen atoms. A non-limiting example of a quaternary ammonium can be a moiety with four organic substituents on a nitrogen atom, which can generally be represented by the formula NR4 + X - where R is any chemically permissible substituent, including acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds, and X is a suitable counterion (or anion), a negatively charged group associated with the positively charged quaternary amine to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F, CI, Br, I), NO3", CIO4", OH", H2PO4", HSO4", sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene- 1 -sulfonic acid-5-sulfonate, ethane- 1 -sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like). "Quaternization" refers to the act of reacting with an R group as described above to form a quaternary ammonium. It will be understood that quaternization of an amino acid of a cyclic peptide compound of the present application means that the NH2on the amino acid residue can be quaternized.

[0260] "Fatty acid" refers to a long aliphatic hydrocarbon chain that is straight chain (i.e., unbranched) or branched, substituted or unsubstituted, and contains at least one carboxyl group at one end, said fatty acid preferably having from 1 to 30 carbon atoms, i.e., "C 1-30 fatty acid", preferably having from 1 to 24 carbon atoms (i.e., C 1-24 fatty acid), further preferably having from 1 to 22 carbon atoms (i.e., C 1-22fatty acids). Non-limiting examples include: lauric acid, myristic acid, stearic acid, palmitic acid, oleic acid, isoleucine, palmitoleic acid, stearic acid, oleic acid, isoleucine, nervonic acid, linoleic acid, a-linolenic acid, g-linolenic acid, arachidonic acid, EPA, DPA, DHA, sebacic acid, C 12 dibasic acids, C 14 dibasic acids, C 16 dibasic acids, C 18 dibasic acids, C 20 dibasic acids, etc. “Heterofatty acid” refers to a fatty acid having at least one heteroatom, i.e., one or more carbon atoms in the fatty acid are replaced with a heteroatom having at least one heteroatom (e.g., nitrogen, oxygen, P(O) m and S(O) n (where m, n are integers from 0-2) non-limiting examples include: 9-hydroxynonanoic acid.

[0261] exemplary C 1-24 fatty acid or C 1-24 heterofatty acid

[0262] “Polyethylene glycol” or “PEG” is a polyether compound of the general formula H-(0-CH2-CH2) n -OH. PEGs are also known as polyethylene oxides (PEO) or polyoxyethylenes (POE), depending on their molecular weight, but PEG tends to refer to oligomers and polymers with a molecular mass of less than 20,000 Da. In some embodiments, PEGs with a molecular weight of 50 to 2,000 or 100 to 500 are used. Different forms of PEGs can also be used, depending on the initiator used in the polymerization process - a common initiator is monofunctional methyl ether PEG or methoxy poly(ethylene glycol) (abbreviated as mPEG). In some embodiments, the PEG is PEG3, PEG4, PEG5, PEG6, PEG7, PEG8, PEG9, PEG10, or PEG11. In some embodiments, the PEG linker is

[0263] The term “carboxyl” refers to -COOH, “Ac” refers to “acetyl”;

[0264] The term “linker” broadly refers to a chemical structure that is capable of linking or binding two peptide monomer subunits together to form a dimer.

[0265] A "dimer" broadly refers to a peptide comprising two or more monomeric subunits. Certain dimers comprise two DRPs. Dimers of the present invention include homodimers and heterodimers. The monomeric subunits of a dimer can be linked at their C-terminus or N-terminus, or they can be linked via internal amino acid residues. The monomeric subunits of a dimer can be linked through the same site, or each can be linked through a different site (e.g., C-terminus, N-terminus, or internal site).

[0266] The term "cyclization" or "forming a peptide loop" refers to a reaction in which a portion of a polypeptide molecule is linked to another portion of the polypeptide molecule to form a closed loop, or a portion of a polypeptide molecule is linked to several other portions of the polypeptide molecule to form multiple closed loops, such as by forming a disulfide bridge or other similar bond, or by a linker.

[0267] A "derivative" or "analog" refers to a product derived from a compound in which a hydrogen atom or a group of atoms is replaced by another atom or group of atoms. It will be appreciated that amino acid analogs of the peptide compounds as defined herein are within the scope of the present invention. Examples of such suitable modified amino acid derivatives include one or more modifications selected from the group consisting of: N-terminal and / or C-terminal modifications; replacement of one or more amino acid residues with one or more non-natural amino acid residues (e.g., replacement of one or more polar amino acid residues with one or more isosteric or isoelectronic amino acids; replacement of one or more non-polar amino acid residues with other non-natural isosteric or isoelectronic amino acids); addition of a spacer group; replacement of one or more oxidation-sensitive amino acid residues with one or more oxidation-resistant amino acid residues; replacement of one or more amino acid residues with alanine, replacement of one or more L amino acid residues with one or more D amino acid residues; N-alkylation of one or more amide bonds in a bicyclic peptide ligand; replacement of one or more peptide bonds with a surrogate bond; peptide backbone length modification; replacement or substitution of the hydrogen on the alpha carbon of one or more amino acid residues with another chemical group, modification of amino acids such as glycine, alanine, phenylalanine, cysteine, lysine, glutamic acid / aspartic acid, and tyrosine, etc. with suitable amine, thiol, carboxylic acid, and phenol reactive reagents to functionalize the amino acid, and introduction or replacement of amino acids to introduce orthogonal reactivity suitable for functionalization, for example, amino acids bearing an azide or alkyne group to allow functionalization with alkyne or azide bearing moieties, respectively.

[0268] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. In the chemical arts, a dash ("-") is used to indicate a point of attachment in a chemical name or formula. Chemical groups can be depicted with or without one or more dashes without loss of generality. A wavy line drawn by a line in a structure indicates a point of attachment of a group. Dashed lines indicate optional bonds. Unless chemically or structurally necessary, the order of a chemical group's writing or the point of attachment of a chemical group to the rest of the molecule does not indicate or imply directionality. For example, the group "-SO2CH2-" is equivalent to "-CH2SO2-," which can be attached in either direction. Similarly, an "arylalkyl" group, for example, can be attached to the rest of the molecule at either the aryl or alkyl portion of the group. As in "Cu-v" or (Cu-Cv), indicates that the group has from u to v carbon atoms. For example, "C1-C6alkyl" and "C1-C6alkyl" both mean that the alkyl group has from 1 to 6 carbon atoms. 1-6 "alkyl" and "C1-C6alkyl" both mean that the alkyl group has from 1 to 6 carbon atoms.

[0269] Throughout this specification, unless a naturally occurring amino acid is referred to by a common name (e.g., alanine, arginine, etc.), it is designated by a conventional three letter or single letter abbreviation (e.g., for alanine, Ala or A; for arginine, Arg or R, etc.). Unless otherwise indicated, the three letter and single letter abbreviations for the amino acids refer to the L-isomer of the amino acid in question. The term "L-amino acid" as used herein refers to the "L" isomeric form of a peptide, in contrast the term "D-amino acid" refers to the "D" isomeric form of a peptide (e.g., Dasp, (D)Asp or D-Asp; Dphe, (D)Phe or D-Phe). A D isomeric form of an amino acid residue can be substituted for any L-amino acid residue so long as the peptide retains the desired functionality. When referred to using the single letter abbreviation, the D-amino acid can be denoted by the lower case letter by convention.

[0270] In the case of unusual amino acids or non-naturally occurring amino acids, unless they are referred to by a common name (e.g., sarcosine, ornithine, etc.), the commonly used three character code or four character code is used for their residues, including, Sar or Sarc (sarcosine, i.e., N-methylglycine), Aib (a-aminoisobutyric acid), Dab (2,4-diaminobutyric acid), Dapa (2,3-diaminopropionic acid), γ-Glu (γ-glutamic acid), Gaba (γ-aminobutyric acid), β-Pro (pyrrolidine-3-carboxylic acid), and 8Ado (8-amino-3,6-dioxaoctanoic acid), Abu (2-aminobutyric acid), βhPro (β-homoproline), βhPhe (β-homophenylalanine), and Bip (β,β diphenylalanine), as well as Ida (iminodiacetic acid).

[0271] Unless otherwise indicated, all amino acids are used in the L-configuration.

[0272] The common names and three letter codes of some common amino acids are shown in the following table:

[0273] The term "pharmaceutically acceptable" means, within the scope of sound medical judgment, that the material is suitable for use with patients with no undue toxicity, irritation, allergic response, and the like, commensurate with a reasonable benefit / risk ratio, and effective for its intended use.

[0274] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application which is safe and effective for use in a mammal, and possesses the desired biological activity.

[0275] The term "pharmaceutical composition" refers to a composition comprising one or more compounds of the present application, or a physiologically / pharmaceutically acceptable salt or prodrug thereof, and other components such as a physiologically / pharmaceutically acceptable carrier or excipient. The purpose of a pharmaceutical composition is to facilitate administration of the active agent to an organism, and to facilitate absorption of the active agent into the organism to exert a biological activity.

[0276] The term "pharmaceutically acceptable carrier" means a material that is not biologically or otherwise undesirable, i.e., the material can be administered to an organism without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the composition in which it is contained. The term "pharmaceutically acceptable carrier" also refers to a nontoxic carrier that does not interfere with the effectiveness of the biological activity of the active compound. The term "pharmaceutically acceptable carrier" includes, but is not limited to, excipients, sweetening agents, diluents, preservatives, dyes / colorants, flavoring agents, surfactants, wetting agents, dispersing agents, suspending agents, breakers, stabilizers, solvents, or emulsifiers.

[0277] The terms "administration" or "administering" or the like refer to methods allowing the delivery of a compound or composition to the desired site of biological action. These methods include, but are not limited to, oral or parenteral (including intracerebroventricular, intravenous, subcutaneous, intraperitoneal, intramuscular, intravascular injection or infusion), topical, rectal, etc. In particular, injection or oral administration.

[0278] As used herein, the term "treatment" includes alleviating, abating or ameliorating a disease or condition, preventing additional symptoms, ameliorating or preventing metabolic factors that contribute to a symptom, inhibiting the disease or condition, e.g., arresting the development of a disease or condition, relieving the symptoms of a disease or condition, facilitating remission of a disease or condition, or stopping the symptoms of a disease or condition, and extends to preventing. "Treatment" also includes achieving a therapeutic benefit and / or a prophylactic benefit. By therapeutic benefit is meant eradication or amelioration of the symptoms of the disease in question. Also, therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological parameters associated with the disease in question, although the patient can still be afflicted with the disease. By prophylactic benefit is meant that the composition is used in a patient who may

[0279] The term "active ingredient," "therapeutic agent," "active substance," or "active agent" refers to a chemical entity that is effective in treating or preventing a disorder, disease, or condition of interest. The term "neuropsychiatric disorder" refers to the general class of neurological and psychiatric disorders, including neurological and / or psychiatric disorders.

[0280] The term "effective amount," "therapeutically effective amount," or "prophylactically effective amount" with respect to a drug, pharmaceutical agent, or active ingredient means an amount of the drug or agent that is sufficient to provide a desired effect, but is acceptable in terms of side effects. Determination of an effective amount is dependent on the age and general condition of the individual, as well as the particular active substance, and an effective amount in a case can be determined by a person of ordinary skill in the art according to routine testing.

[0281] An "individual" as used herein includes a human or non-human animal. Exemplary human individuals include a human individual (referred to as a patient) who has a disease (e.g., a disease described herein) or a normal individual. "Non-human animals" in the present application include all vertebrates, e.g., non-mammals (e.g., birds, amphibians, reptiles) and mammals, e.g., non-human primates, domestic animals, and / or laboratory models (e.g., sheep, dog, cat, cow, pig, etc.).

[0282] The term "room temperature" refers to a temperature from 10 °C to 40 °C. In some embodiments, "room temperature" refers to a temperature from 15 °C to 30 °C; in other embodiments, "room temperature" refers to a temperature from 18 °C to 25 °C.

[0283] "Equivalent" or its abbreviation "eq" is the equivalent relationship in the chemical reaction, based on the amount of the basic raw material used in each step (1 equivalent), and the equivalent amount of other raw materials required.

[0284] The "*" in the chemical structure of the examples indicates a single configuration R or S.

[0285] The following detailed description of the application is intended to illustrate, but not limit, the embodiments of the application, so that others can better understand the technical solutions of the application, its principles and its practical applications, so that others can modify and implement the application in many forms to best adapt it to the requirements of specific uses. DETAILED DESCRIPTION

[0286] The content of the application will be described in detail below by way of examples. The specific conditions not specified in the examples are carried out according to the conventional conditions of experimental methods. The examples are given to better illustrate the content of the application, but should not be understood as limiting the content of the application to the examples. The person skilled in the art can make non-essential improvements and adjustments to the embodiments according to the above content of the application, which still belong to the protection scope of the application.

[0287] Detection method: The structure of the compound is determined by mass spectrometry (MS).

[0288] MS determination is carried out by Agilent 6120B (ESI) and Agilent 6120B (APCI);

[0289] HPLC determination is carried out by Agilent 1260DAD high pressure liquid chromatograph (Zorbax SB-C18 100x4.6mm, 3.5μM);

[0290] Abbreviation explanation: DCM: dichloromethane; DMF: N,N-dimethylformamide; DIEA: N,N-diisopropyl ethylamine; MeOH: methanol; TFA: trifluoroacetic acid; DMSO: dimethyl sulfoxide; DIC: N,N'-diisopropyl carbodiimide; HOBT: 1-hydroxybenzotriazole; HOAT: N-hydroxy-7-azabenzotriazole.

[0291] EXAMPLE

[0292] The specific conditions not specified in the examples are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not specified by the manufacturer, which are conventional products that can be obtained by purchase. The proportions or percentages used in this article are by weight, unless otherwise specified.

[0293] Intermediate 1:

[0294] First step: Compound 1a (10.0 g, 73.43 mmol) was dissolved in dichloromethane (100 mL), and TiCl4(25.5 g, 134.38 mmol) was added dropwise to the above reaction solution, which was cooled to 0 °C. After 5 min, 1,1-dichloro dimethyl ether (9.4 g, 81.51 mmol) was added dropwise, and the mixture was stirred at 0 °C for 3 h. After the reaction was completed, 200 mL of water was added to the reaction system, and the resulting solution was extracted with dichloromethane, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Column chromatography (EA: PE = 1:4) gave compound 1b (5.58 g, 46.3%).

[0295] LC-MS (ESI): m / z = 163.2 [M+H] + .

[0296] Second step: Compound 1b (3.0 g, 18.27 mmol), tert-butyl (2-bromoethyl)carbamate (4.9 g, 21.91 mmol), potassium carbonate (5.1 g, 36.54 mmol), and sodium iodide (0.8 g, 5.47 mmol) were dissolved in N,N-dimethylformamide (20 mL) and stirred at 25 °C for 16 h. After the reaction was completed, 100 mL of water was added, and the reaction solution was extracted three times with 50 mL of ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography (EA: PE = 4:1) gave compound 1c (4.9 g, 87%).

[0297] LC-MS (ESI): m / z = 306.1 [M+H] + .

[0298] Third step: (±)Benzyl oxycarbonyl-a-phosphonoglycine trimethyl ester (6.2 g, 18.74 mmol) was dissolved in dichloromethane (100 mL), and after nitrogen replacement, DBU (3.1 g, 20.31 mmol) was added and stirred for 30 min. Compound 1c (4.8 g, 15.62 mmol) was then dissolved in dichloromethane (100 mL) and added dropwise to the reaction, which was stirred overnight. The reaction mixture was diluted with dichloromethane (100 mL) and washed with 5% aqueous citric acid solution (100 mL) and saturated brine (100 mL), then dried over anhydrous sodium sulfate, filtered, and rotary evaporated. Purification by silica gel chromatography (EA: PE = 3:1) gave compound 1d (6.1 g, 76.2%).

[0299] LC-MS (ESI): m / z = 511.1 [M+H] + .

[0300] Fourth Step: Compound 1d (6.0 g, 11.71 mmol) was dissolved in a mixed solution of methanol (50 mL) and dichloromethane (20 mL), (+)-1,2-bis(2S,5S)-2,5-diethylcyclobutylphosphine benzene (cyclooctadiene) rhodium trifluoromethanesulfonate (0.6 g, 0.83 mmol) was added, a high-pressure kettle was filled with hydrogen gas to a pressure of about 4.0 bar, and stirring was performed at room temperature for 3 h. The solid was filtered off, and the reaction solution was concentrated to obtain compound 1e (5.9 g, 98%).

[0301] LC-MS (ESI): m / z = 513.2 [M+H] + .

[0302] Fifth Step: Compound 1e (5.9 g, 11.46 mmol) was dissolved in a mixed solution of methanol (50 mL) and dichloromethane (20 mL), and palladium-carbon (10%, 1 g) was added. Stirring was performed under a hydrogen gas environment overnight, and then the solid was filtered off. The reaction solution was rotary evaporated to obtain compound 1f (3.7 g, 84.5%).

[0303] LC-MS (ESI): m / z = 379.4 [M+H] + .

[0304] Sixth Step: Compound 1f (3.7 g, 9.72 mmol) was dissolved in tetrahydrofuran (25 mL), lithium hydroxide monohydrate (1.2 g, 48.6 mmol) and water (25 mL) were added, and the reaction was stirred at room temperature overnight. Impurities were extracted (15 mL x 3) with ethyl acetate, and then the water phase was adjusted to neutral pH with dilute hydrochloric acid (1N). The product was precipitated, filtered and dried to obtain compound 1g (3.2 g, 89.8%).

[0305] LC-MS (ESI): m / z = 365.2 [M+H] + .

[0306] Seventh Step: Compound 1g (2.7 g, 7.41 mmol) was dissolved in a mixed solution of acetonitrile (25 mL) and water (25 mL), sodium bicarbonate (6.2 g, 74.16 mmol) and 9-fluorenylmethyl-N-succinimidyl carbonate (3.3 g, 9.63 mmol) were added, and the reaction was stirred at room temperature overnight. After the reaction was completed, dilute hydrochloric acid was added dropwise to adjust the pH to neutral, most of the acetonitrile was removed by vacuum concentration at 40°C, and the solid was collected by filtration to obtain the crude product. Purification was performed by silica gel column chromatography (DCM:MeOH = 10:1) to obtain intermediate 1 (2.7 g, 62.1%).

[0307] LC-MS (ESI): m / z = 587.2 [M+H] + .

[0308] 1 H NMR (400 MHz, DMSO-d6) δ 12.65 (s, 1H), 7.88 (d, 2H), 7.67 (s, 2H), 7.41 (d, 2H), 7.36-7.24 (m, 2H), 7.03-6.88 (m, 2H), 6.65 (d, 1H), 4.26-4.09 (m, 4H), 3.91 (s, 2H), 3.28 (s, 2H), 2.99 (d, 1H), 2.90-2.68 (m, 5H), 2.51 (s, 1H), 1.98 (s, 2H), 1.39 (s, 9H).

[0309] Intermediate 4:

[0310] First Step: Intermediate 1 (35.0 g, 71.93 mmol) in dichloromethane (50 mL) was added to a mixture of dichloromethane (100 mL) and trifluoroacetic acid (50 mL) and stirred at room temperature for 1 h. After completion of the reaction, it was concentrated under reduced pressure to get crude Intermediate 4a (35.83 g, 100%).

[0311] LC-MS (ESI): m / z = 487.2 [M+H] + .

[0312] Second Step: Intermediate 4a (26.8 g, 55.08 mmol) and 2-acetyl-5,5-dimethyl-1,3- cyclohexanedione (40.2 g, 220.32 mmol) were added to a mixture of tetrahydrofuran (210 mL) and glacial acetic acid (42 mL) and stirred at 95 °C for 16 h. After completion of the reaction, it was concentrated under reduced pressure and purified by reverse phase column (ACN: H20 = 3:2) to get Intermediate 4 (11.2 g, 45.9%).

[0313] LC-MS (ESI): m / z = 651.3 [M+H] + .

[0314] 1HNMR (400 MHz, DMSO-d6) δ 13.46-13.43 (m, 1H), 12.64 (s, 1H), 7.88-7.86 (d, 2H), 7.68-7.64 (m, 3H), 7.43-7.38 (m, 2H), 7.33-7.27 (m, 2H), 7.01-6.99 (m, 1H), 6.69-6.67 (d, 1H), 4.22-4.20 (m, 2H), 4.09-4.03 (m, 4H), 3.82-3.80 (m, 4H), 3.02-2.97 (m, 1H), 2.84-2.79 (m, 5H), 2.53 (s, 3H), 2.27 (s, 4H), 0.94 (s, 6H).

[0315] Intermediate 5:

[0316] First step: Compound 5a (5.0 g, 20.07 mmol) was dissolved in dichloromethane (50 mL), and triethylamine (4.1 g, 40.13 mmol) and 4-dimethylaminopyridine (245.2 mg, 2.01 mmol) were added in turn. The mixture was stirred in an ice bath for 5 min, and p-toluenesulfonyl chloride (4.6 g, 24.08 mmol) was added in portions. The reaction was allowed to reach room temperature and was carried out for 2 h. After the reaction was completed, 50 mL of water was added to the reaction system, and the resulting solution was extracted with dichloromethane, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM:MeOH = 10:1) to obtain compound 5b (7.4 g, 91.5%).

[0317] LC-MS (ESI): m / z = 304.1 [M-Boc+H] + .

[0318] Second step: Compound 5b (5.0 g, 12.40 mmol) was dissolved in acetonitrile (50 mL), and potassium carbonate (3.4 g, 24.80 mmol) and N-methylbenzylamine (2.3 g, 18.60 mmol) were added in turn. The mixture was heated to 80°C and reacted for 10 h. After the reaction was completed, 100 mL of water was added to the reaction system, and the resulting solution was extracted with ethyl acetate, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography (DCM:MeOH = 10:1) to obtain compound 5c (3.5 g, 80.1%).

[0319] LC-MS (ESI): m / z = 353.2 [M+H] + .

[0320] Third step: Take compound 5c (3.0 g, 8.52 mmol) dissolved in dichloromethane (30 mL), drop 10 mL of trifluoroacetic acid in ice bath, stir in ice bath, 0.5 hours. After the reaction is completed, adjust the solution to be alkaline with saturated sodium bicarbonate solution, add 20 mL of water, and extract the resulting solution with dichloromethane, dry with anhydrous sodium sulfate, and concentrate under reduced pressure to obtain intermediate 5 (1.8 g, 83.8%).

[0321] LC-MS (ESI): m / z = 253.2 [M+H] +

[0322] Intermediate 6:

[0323] First step: Add 6a (5 mmol, 5 g, sub: 1 mmol / g) and dichloromethane to the reactor, swell the resin for 5 min, then add Fmoc-AEEA-OH (7.5 mmol, 2.9 g), N,N-diisopropylethylamine (15 mmol, 2.6 ml) in turn, react at room temperature for 1 h, then add methanol (5 ml) and react for 30 min, dry the solvent and wash the resin with DMF 3 times to obtain 6b.

[0324] Second step: Add 20% piperidine / DMF to 6b, dry the solvent after reacting for 30 min and wash the resin with DMF 5 times to obtain 6c, and monitor the reaction with ninhydrin.

[0325] Third step: Add Fmoc-Glu-OtBu (10 mmol, 4.25 g), DMF (20 ml), 1-hydroxybenzotriazole (10 mmol, 1.35 g), N,N'-diisopropylcarbodiimide (10 mmol, 1.57 ml) to 6c in turn, react at room temperature for 1.5 h, and monitor the reaction with ninhydrin after the reaction is completed, dry the solvent and wash the resin with DMF 3 times to obtain 6d.

[0326] Fourth step: Add 20% piperidine / DMF to 6d, dry the solvent after reacting for 30 min and wash the resin with DMF 5 times to obtain 6e, and monitor the reaction with ninhydrin.

[0327] Fifth step: Add octadecanedioic acid mono-tert-butyl ester (10 mmol, 3.7 g), DMF (20 ml), 1-hydroxybenzotriazole (10 mmol, 1.35 g), N,N'-diisopropylcarbodiimide (10 mmol, 1.57 ml) to 6e in turn, react at room temperature for 1 h, and monitor the reaction with ninhydrin after the reaction is completed, dry the solvent and wash the resin with DMF 3 times, methanol 2 times, dichloromethane 1 time, methanol 2 times, and dry to obtain 6f.

[0328] Step 6: To 6f (7.3 g) was added 30% trifluoroethanol in dichloromethane (70 ml) and stirred at room temperature for 1 h. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain 6g (2.6 g, 74% yield)

[0329] LCMS (ESI): m / z = 701.7 [M+H] + .

[0330] Step 7: To 6g (1.9 g, 2.7 mmol) and intermediate 5 (0.7 g, 2.7 mmol) was added dichloromethane (20 mL) followed by HATU (0.87 g, 2.7 mmol) and DIPEA (0.7 g, 5.4 mmol) and stirred at room temperature for 16 h. The reaction mixture was extracted with dichloromethane (100 mL) and water (100 mL). The dichloromethane layer was concentrated to dryness and purified by column chromatography to obtain compound 6h (1.5 g, 60%).

[0331] Step 8: To 6h (1.5 g, 1.6 mmol) was added methanol (20 mL) followed by 10% palladium on carbon (150 mg) and stirred at room temperature for 16 h. The reaction mixture was filtered through celite and concentrated to dryness to obtain intermediate 6 (1.3 g, 96%).

[0332] LCMS (ESI): m / z = 845.6 [M+H] + .

[0333] Intermediate 13:

[0334] Step 1: To the reactor was added 6a (2 mmol, 2 g, sub: 1 mmol / g) and dichloromethane, the resin was swelled for 5 min, followed by the addition of Fmoc-AEEA-OH (3 mmol, 1.2 g), N,N-diisopropylethylamine (6 mmol, 1.0 ml) and stirred at room temperature for 1 h. Methanol (5 ml) was added and stirred for 30 min. The solvent was drained and the resin was washed with DMF for 3 times to obtain 13b.

[0335] Step 2: To 13b was added 20% piperidine in DMF and stirred for 30 min. The solvent was drained and the resin was washed with DMF for 5 times to obtain 13c. The reaction was monitored by ninhydrin test.

[0336] Step 3: To 13c was added Fmoc-AEEA-OH (4 mmol, 1.5 g), DMF (20 ml), 1-hydroxybenzotriazole (4 mmol, 0.54 g), N,N'-diisopropylcarbodiimide (4 mmol, 0.62 ml) and stirred at room temperature for 1.5 h. The reaction was monitored by ninhydrin test. The solvent was drained and the resin was washed with DMF for 3 times to obtain 13d.

[0337] Fourth step: To 13d, add 20% piperidine / DMF, after 30 minutes of reaction, dry the solvent and wash the resin with DMF for 5 times, to get 13e, ninhydrin monitor the reaction.

[0338] Fifth step: To 13e, add Fmoc-Glu-OtBu (4 mmol, 1.7 g), DMF (20 ml), 1- hydroxybenzotriazole (4 mmol, 0.54 g), N, N'-diisopropylcarbodiimide (4 mmol, 0.62 ml) in sequence, react for 1.5 h at room temperature, after the reaction is completed, dry the solvent and wash the resin with DMF for 3 times, to get 13f.

[0339] Sixth step: To 13f, add 20% piperidine / DMF, after 30 minutes of reaction, dry the solvent and wash the resin with DMF for 5 times, to get 13g, ninhydrin monitor the reaction.

[0340] Seventh step: To 13g, add octadecanedioic acid mono-tert-butyl ester (4 mmol, 1.48 g), DMF (20 ml), 1-hydroxybenzotriazole (4 mmol, 0.54 g), N, N'- diisopropylcarbodiimide (4 mmol, 0.62 ml) in sequence, react for 1.5 h at room temperature, after the reaction is completed, dry the solvent and wash the resin with DMF for 3 times, methanol for 2 times, dichloromethane for 1 time, methanol for 2 times, dry to get 13h.

[0341] Eighth step: To 13h (3.2 g), add 30% trifluoroethanol / dichloromethane solution (32 ml), after 1 h of reaction at room temperature, filter, collect the filtrate and concentrate under reduced pressure to get 13i (1.0 g)

[0342] Ninth step: Add 13i (1.0 g, 1.18 mmol) into DCM (10 mL), then add N-Cbz-N- methylethylenediamine (0.246 g, 1.18 mmol), HATU (0.45 g, 1.18 mmol), DIPEA (0.3 g, 2.36 mmol). React for 2 h at room temperature, add water and dichloromethane for extraction, concentrate the dichloromethane phase to dryness, purify by column chromatography to get 13j (600 mg, yield: 50%)

[0343] Tenth step: Add 13j (0.6 g, 0.587 mmol) into methanol (10 mL), then add 10% palladium carbon (0.1 g). React for 18 h at room temperature under hydrogen atmosphere, filter and concentrate to dryness to get intermediate 13 (0.47 g, yield: 71%)

[0344] LCMS m / z = 902.8 [M+H] + .

[0345] Intermediate 14:

[0346] First step: Take compound 14a (12.7 g, 29.8 mmol) in a flask, dissolved in dichloromethane (120 mL), sequentially add raw material 2-(2-(2-aminoethoxy)ethoxy)ethyl carbamate tert-butyl (8.5 g, 35.7 mmol), DIEA (7.7 g, 59.6 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (6.9 g, 35.7 mmol) is added in batches under ice bath, and the reaction is heated to room temperature for 10 h. Add water (60 mL) to the reaction system, separate the layers, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and separate by column chromatography (DCM:MeOH=10:1) to obtain compound 14b (19 g, 97%).

[0347] LCMS (ESI): m / z = 584.0 [M+H] + .

[0348] Second step: Take compound 14b (19.0 g, 28.9 mmol) in a flask, dissolved in dichloromethane (200 mL), drop trifluoroacetic acid (100 mL) at room temperature, stir at room temperature for 1 h, heat to 45°C for 10 min, after the reaction is completed, directly concentrate under reduced pressure to obtain a yellow oily liquid, and purify by reverse phase column (ACN:H2O=50:50) to obtain compound 14c (9.8 g, 55%).

[0349] LCMS (ESI): m / z = 500.0 [M+H] + .

[0350] Third step: Take compound 14c (2.4 g, 3.9 mmol) in a flask, add ethanol (25 mL) to dissolve, add 2-acetyl-5,5-dimethyl-1,3-cyclohexanedione (1.7 g, 7.8 mmol), drop trifluoroacetic acid (0.3 mL), heat to 90°C for 10 h. After the reaction is completed, concentrate under reduced pressure, add dichloromethane (100 mL) and water (40 mL) to extract, dry the organic phase with anhydrous sodium sulfate, concentrate under reduced pressure, and separate by column chromatography (DCM:MeOH=15:1) to obtain intermediate 14 (2.2 g, 85%).

[0351] LCMS (ESI): m / z = 664.4 [M+H] + .

[0352] Intermediate 29:

[0353] First step: Compound 29a (10 g, 37.7 mmol) and pyridine chlorochromate (12.2 g, 56.5 mmol) were added into dichloromethane (150 ml) and reacted at room temperature for 16 h. After the reaction was completed, the mixture was filtered and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 29b (8 g, 80.6%).

[0354] Second step: Compound 29c (10 g, 44.82 mmol) and triphenylphosphine (14.11 g, 53.79 mmol) were added into acetonitrile (150 ml) and reacted at 85°C for 35 h. The reaction mixture was directly concentrated under reduced pressure and purified by silica gel column chromatography to obtain 29d (20 g, 92%).

[0355] Third step: 29d (20 g, 41 mmol) was added into dry THF (150 ml), and after N2 replacement, the temperature was lowered to 0°C. NaHMDS (41 mmol) was slowly added, and the mixture was stirred at 0°C for 20 min. A solution of 29b (7.2 g, 27 mmol) in tetrahydrofuran (30 ml) was slowly added dropwise, and the mixture was reacted at 0°C for 1 h. Water (100 ml) was added, and the mixture was extracted with ethyl acetate (100 ml x 2). The combined organic phase was washed with saturated sodium chloride solution, and the organic phase was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 29e (5.5 g, 52.5%).

[0356] Fourth step: 29e (5.5 g, 14.2 mmol) was added into ethyl acetate (70 ml), and 10% Pd / C (1 g) was added. The mixture was reacted at room temperature for 16 h. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain 29f (5.5 g, 99.5%).

[0357] Fifth step: 29f (5.4 g, 13.8 mmol), dibenzylphosphinic acid (4 g, 15.2 mmol), tetraethylammonium iodide (3.56 g, 13.8 mmol), and cesium carbonate (9 g, 27.7 mmol) were added into DMF (60 ml), and the mixture was reacted at 60°C for 16 h. The mixture was cooled to room temperature, water (300 ml) was added, and the mixture was extracted with ethyl acetate (100 ml x 3). The organic phase was washed with saturated sodium chloride solution, and the mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain 29g (4 g, 50.62%).

[0358] Sixth step: 29g (4 g, 7 mmol) and lithium hydroxide (0.84 g, 35 mmol) were added into a mixed solvent of methanol (30 ml) and water (10 ml), and the mixture was reacted at room temperature for 3 h. Most of the methanol was removed by concentration under reduced pressure. The mixture was adjusted to pH = 4-5 with 6 mol / L hydrochloric acid, and a large amount of solid was precipitated. The solid was filtered, washed with water, and dried to obtain intermediate 29 (3.7 g, 94.82%).

[0359] LC-MS (ESI): m / z = 559.4 [M+H] + .

[0360] 1 H NMR (400 MHz, DMSO-d6) δ 12.07 (s, 1H), 7.41 - 7.26 (m, 10H), 5.04 - 4.92 (m, 4H), 2.17 (t, 2H), 1.83 - 1.70 (m, 2H), 1.50 - 1.39 (m, 4H), 1.29 - 1.18 (m, 28H).

[0361] Intermediate 35:

[0362] First step: Compound 35a (10 g, 95.11 mmol) and benzyl bromide (32.6 g, 190.22 mmol) were dissolved in acetonitrile (200 mL), then potassium carbonate (39.38 g, 285.33 mmol) was added, stirred at 80 °C for 4 h, after the reaction was completed, cooled, filtered off the solid, washed with acetonitrile and spin dry the solvent, then separated and purified using a silica gel column (PE:EA = 1:1) to obtain compound 35b (26 g, 95.8%).

[0363] LC-MS (ESI): m / z = 286.1 [M+H] + .

[0364] Second step: Compound 35b (20 g, 70.16 mmol) was dissolved in dichloromethane (200 mL), triethylamine (21.3 g, 210.53 mmol) and p-toluenesulfonyl chloride (20 g, 105.26 mmol) were added, then 4-dimethylaminopyridine (856 mg, 7.0 mmol) was added, stirred at room temperature for 3 h, water (500 mL) was added, extracted with dichloromethane (200 mL x 2), the organic phase was combined and concentrated, then separated and purified using a silica gel column (PE:EA = 3:1) to obtain compound 35c (28.8 g, 93.5%).

[0365] LC-MS (ESI): m / z = 440.1 [M+H] + .

[0366] Third Step: Dissolve N-Boc-3-hydroxyazetidine (12 g, 69.3 mmol) in dry DMF (200 mL), place the reaction at 0 °C, slowly add sodium hydride (3.33 g, 83.2 mmol), after the addition, stir for 30 min, then add compound 35c (28.8 g, 65.6 mmol) dissolved in DMF (100 mL) dropwise into the reaction, continue to stir at room temperature for 16 h, add water (1 L), extract with ethyl acetate (200 mL x 3), dry over anhydrous sodium sulfate, combine the organic phase and concentrate, purify by silica gel chromatography column (PE:EA = 1:1) to obtain compound 35d (26.3 g, 91.1%).

[0367] LC-MS (ESI): m / z = 441.2 [M+H] + .

[0368] Fourth Step: Dissolve compound 35d (26 g, 59.1 mmol) in methanol (300 mL), add palladium on carbon (5.2 g, 10%), after replacement with hydrogen, react at room temperature for 48 h, then filter and dry to obtain compound 35e (15 g, 97.2%).

[0369] LC-MS (ESI): m / z = 261.1 [M+H] + .

[0370] Fifth Step: Dissolve compound 35e (15 g, 57.7 mmol) in dichloromethane (150 mL), add 2-acetyl-5,5-dimethyl-1,3-cyclohexanedione (15.75 g, 86.5 mmol), stir at room temperature for 3 h, after the reaction is complete, concentrate, purify by silica gel chromatography column (PE:EA = 1:2) to obtain compound 35f (19.4 g, 79.3%).

[0371] LC-MS (ESI): m / z = 425.2 [M+H] + .

[0372] Sixth Step: Dissolve compound 35f (5 g, 11.8 mmol) in hydrogen chloride dioxane (50 mL, 4M), stir at room temperature for 2 h, directly dry to obtain hydrochloride of compound 35g (4.25 g, 99%).

[0373] LC-MS (ESI): m / z = 325.2 [M+H] + .

[0374] Step 7: Use compound 35g hydrochloride (4.2 g, 11.7 mmol) and fluorenylmethoxycarbonyl-L-glutamic acid 1-tert-butyl ester (5 g, 11.8 mmol) as raw material, refer to the first step of operation method of intermediate 14 to obtain compound 35h (6.5 g, 68.6%).

[0375] LC-MS (ESI): m / z = 732.2 [M+H] + .

[0376] Step 8: Compound 35h (6.5 g, 8.88 mmol) was dissolved in hydrogen chloride dioxane (65 mL, 4M) and stirred at room temperature for 2 h. After rotary evaporation, the product was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain intermediate 35 (5 g, 83.3%).

[0377] LC-MS (ESI): m / z = 676.2 [M+H] + .

[0378] Intermediate 36:

[0379] Step 1: Compound 36a (5.0 g, 21.80 mmol), rhodium dimeric acetate (193 mg, 0.44 mmol) were dissolved in super dry dichloromethane (50 mL) under nitrogen atmosphere. The reaction mixture was cooled to 0 °C, and ethyl diazoacetate (3.73 g, 32.71 mmol) dissolved in super dry dichloromethane (10 mL) was added dropwise. After the addition was completed, the reaction mixture was stirred at room temperature for 16 h. After the reaction was completed, the crude product was obtained by concentration. The crude product was purified to obtain compound 36b (5 g, 73%).

[0380] LC-MS (ESI): m / z = 216.2 [M-100+H] + .

[0381] Step 2: Compound 36b (5.0 g, 15.85 mmol) was dissolved in super dry tetrahydrofuran (50 mL). Lithium aluminum hydride (1.5 g, 39.63 mmol) was added to the above reaction solution at 0 °C. The reaction mixture was stirred at room temperature for 2 h. After the reaction was completed, sodium hydroxide aqueous solution was slowly added at 0 °C for quenching. Ethyl acetate (50 mL x 3) was added for extraction. The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified to obtain compound 36c (4 g, 92%).

[0382] LC-MS (ESI): m / z = 174.1 [M-100+H] + .

[0383] Step 3: Compound 36c (4.0 g, 14.65 mmol), methylsulfonic anhydride (3.06 g, 17.58 mmol), triethylamine (2.96 g, 29.30 mmol), 4-dimethylaminopyridine (358 mg, 2.93 mmol) were dissolved in dichloromethane (50 mL) and stirred at room temperature for 16 h. After the reaction was completed, water (20 mL) was added and extracted with dichloromethane (30 mL x 3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified to give compound 36d (3.5 g, 68%).

[0384] LC-MS (ESI): m / z = 296.1 [M-56+H] + .

[0385] Step 4: Compound 36d (3.5 g, 9.96 mmol) was dissolved in super dry N,N- dimethylformamide (35 mL), and sodium azide (1.29 g, 19.92 mmol) was added. The reaction was carried out at 90 °C for 16 h. After the reaction was completed, water (20 mL) was added and extracted with ethyl acetate (30 mL x 3). The combined organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The crude product was purified to give compound 36e (2.57 g, 86%).

[0386] LC-MS (ESI): m / z = 199.3 [M-100+H] + .

[0387] Step 5: Compound 36e (2.0 g, 6.70 mmol) was dissolved in methanol (20 mL), and Pd / C (400 mg) was added. The reaction was carried out at room temperature for 16 h. After the reaction was completed, the reaction solution was directly filtered and concentrated under reduced pressure to give compound 36f (1.6 g, 88%).

[0388] LC-MS (ESI): m / z = 273.2 [M+H] + .

[0389] Step 6: Compound 36f (1.6 g, 5.87 mmol) was dissolved in dichloromethane (20 mL), and 2-acetyl-5,5-dimethyl-1,3-cyclohexanedione (2.14 g, 11.75 mmol) was added. The reaction was carried out at room temperature for 16 h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure and purified by silica gel column chromatography to give compound 36g (1.55 g, 60%).

[0390] LC-MS (ESI): m / z = 437.3 [M+H] + .

[0391] Seventh step: 36g (1.55g, 3.55mmol) was dissolved in dichloromethane (15 mL), then trifluoroacetic acid (3 mL) was added, and the reaction was carried out at room temperature for 3h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to obtain compound 36h (1.1g, 92%).

[0392] LC-MS (ESI): m / z = 337.1 [M+H] + .

[0393] Eighth step: 36h (1.1g, 3.27mmol) was dissolved in N,N-dimethylformamide (10 mL), and N-Fmoc-L-glutamic acid-1-tert-butyl ester (1.39g, 3.27mmol), 2-(7-azobenzenesulfonyl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.86g, 4.09mmol), N,N-diisopropyl ethylamine (1.69g, 13.08mmol) were added in turn, and the reaction was carried out at room temperature for 5h. After the reaction was completed, water (20 mL) was added, and the combined organic phase was extracted with ethyl acetate (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product, which was purified to obtain compound 36i (1.25g, 51%).

[0394] LC-MS (ESI): m / z = 744.5 [M+H] + .

[0395] Ninth step: 36i (1.25g, 1.68mmol) was dissolved in dichloromethane (15 mL), then trifluoroacetic acid (3 mL) was added, and the reaction was carried out at room temperature for 3h. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure, and then purified by silica gel column chromatography to obtain intermediate compound 36 (0.9g, 82%).

[0396] LC-MS (ESI): m / z = 688.5 [M+H] + .

[0397] 1H NMR (400 MHz, CDC13) δ 13.51 (s, 1H), 7.76 (d, 2H), 7.62-7.56 (m, 2H), 7.40 (t, 2H), 7.34-7.28 (m, 2H), 6.14 (s, 1H), 4.69-4.54 (m, 1H), 4.42-4.28 (m, 2H), 4.26-4.18 (m, 2H), 3.82 (d, 1H), 3.65-3.50 (m, 6H), 3.16-2.91 (m, 2H), 2.72-2.58 (m, 2H), 2.57 (s, 3H), 2.39 (s, 4H), 2.34-2.21 (m, 1H), 2.06-1.96 (m, 1H), 1.87-1.72 (m, 3H), 1.61-1.51 (m, 2H), 1.26-1.07 (m, 2H), 1.03 (s, 6H).

[0398] Intermediate 38:

[0399] First Step: Compound 30a (1.2 g, 2.8 mmol) and tert-butyl (17-amino-3,6,9,12,15- pentaoxahexadecyl)carbamate (1.0 g, 2.8 mmol) were added into dichloromethane (20 ml), followed by the addition of HATU (1.2 g, 3.36 mmol) and DIEA (0.5 g, 4.2 mmol), and the reaction was allowed to proceed at room temperature for 2 h. The reaction was directly concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel to give 38b (2.0 g, 90%).

[0400] Second Step: 38b (2.0 g, 2.5 mmol) was added into dichloromethane (40 ml), followed by the addition of trifluoroacetic acid (12 ml), and the reaction was allowed to proceed at room temperature for 3 h. The reaction was directly concentrated under reduced pressure to give crude 38c (3.3 g).

[0401] LC-MS (ESI): m / z = 632.4 [M+H] + .

[0402] Third Step: 38c (3.3 g), 2-(1-hydroxyethylidene)-5,5-dimethylcyclohexane-1,3-dione (1.9 g, 10.4 mmol) and triethylamine (1.0 g, 10.4 mmol) were added into dichloromethane (40 ml), and the reaction was allowed to proceed at room temperature for 3 h. The reaction was directly concentrated under reduced pressure, and the residue was purified by column chromatography on silica gel to give intermediate 38 (1.8 g, 72%).

[0403] LC-MS (ESI): m / z = 796.6 [M+H] + .

[0404] Intermediate 40:

[0405] First Step: N-BOC-piperazine (10.2 g, 54.6 mmol), compound 35c (12 g, 27.3 mmol) and sodium bicarbonate (6.9 g, 81.9 mmol) were dissolved in dry acetonitrile (200 mL) and reacted at 50 °C overnight. After filtration, the filtrate was concentrated and purified by silica gel chromatography (DCM:MeOH = 10:1) to give compound 40a (6.5 g, 52.5%).

[0406] LC-MS (ESI): m / z = 454.3 [M+H] + .

[0407] Second Step: Compound 40a (2.0 g, 4.4 mmol) and palladium on carbon (0.5 g, 10%) were used as raw materials to give compound 40b (1.2 g, 99.5%) according to the operation method of the fourth step of Intermediate 35.

[0408] LC-MS (ESI): m / z = 274.2 [M+H] + .

[0409] Third Step: Compound 40b (1.2 g, 4.4 mmol) and 2-acetyl-5,5-dimethyl-1,3- cyclohexanedione (1.2 g, 6.6 mmol) were used as raw materials to give compound 40c (1.6 g, 84.2%) according to the operation method of the fifth step of Intermediate 35.

[0410] LC-MS (ESI): m / z = 438.4 [M+H] + .

[0411] Fourth Step: Compound 40c (1.6 g, 3.7 mmol) was used as a raw material to give the crude product of compound 40d (1.6 g, 100%) according to the operation method of the sixth step of Intermediate 35, which was directly used in the next step.

[0412] LC-MS (ESI): m / z = 338.3 [M+H] + .

[0413] Fifth Step: Compound 40d hydrochloride (1.6 g, 3.7 mmol) and fluorenylmethoxycarbonyl-L-glutamic acid 1-tert-butyl ester (1.6 g, 3.7 mmol) were used as raw materials to give compound 40e (2.1 g, 77.7%) according to the operation method of the first step of Intermediate 14.

[0414] LC-MS (ESI): m / z = 745.4 [M+H] + .

[0415] Step 6: Compound 40e (2.1 g, 2.8 mmol) was dissolved in dichloromethane (20 mL), trifluoroacetic acid (6 mL) was added, and the reaction was stirred at room temperature for 2 h. After concentration, the intermediate 40 (1.7 g, 89.4%) was obtained by neutral condition reverse phase purification.

[0416] LC-MS (ESI): m / z = 689.5 [M+H] + .

[0417] Intermediate 41:

[0418] Step 1: Compound 41a (5 g, 24.39 mmol) was dissolved in dichloromethane, triethylamine (7.41 g, 73.17 mmol) and p-toluenesulfonyl chloride (6.99 g, 36.59 mmol) were added, and the mixture was stirred at room temperature for 3 h. After the reaction was completed, water was added for dilution, dichloromethane was used for extraction, the organic phase was collected, anhydrous sodium sulfate was used for drying, filtration was performed, concentration was performed, and fast purification by silica gel column chromatography was performed to obtain compound 41b (8.49 g, 97%).

[0419] LCMS m / z = 360.2 [M+H] +

[0420] Step 2: Compound 41c (1 g, 5.75 mmol) was placed in a 50 ml round-bottom flask, 4N hydrogen chloride-1,4-dioxane (20 ml) was added, and stirring was performed at room temperature for 2 h. After the reaction was completed, direct concentration was performed to obtain compound 41d hydrochloride salt, which was directly used in the next step.

[0421] Step 3: The compound 41d hydrochloride salt in the previous step was placed in a 50 ml round-bottom flask, dissolved in dichloromethane (20 ml), triethylamine (0.7 g, 6.9 mmol) and DDE-OH (1.57 g, 8.63 mmol) were added, and stirring was performed at room temperature overnight. After the reaction was completed, water was added for dilution, dichloromethane was used for extraction, the organic phase was collected, anhydrous sodium sulfate was used for drying, filtration was performed, concentration was performed, and fast purification by silica gel column chromatography was performed to obtain compound 41e (1.2 g, 88%).

[0422] LCMS m / z = 239.2 [M+H] +

[0423] Step 4: Compound 41b (1 g, 2.79 mmol) and compound 41e (0.66 g, 2.79 mmol) were added to N,N-dimethylformamide (10 mL), cesium carbonate (1.82 g, 5.57 mmol) was added, the reaction was stirred in a 100°C oil bath kettle for 3 h, TLC was used to monitor the completion of the reaction, water was added for quenching, ethyl acetate was used for extraction, the organic phase was collected, the organic phase was concentrated, and then compound 41f (0.53 g, 45%) was obtained by using a silica gel chromatographic column for purification.

[0424] LCMS m / z = 426.3 [M+H] +

[0425] Step 5: Compound 41f (0.53 g, 1.25 mmol) was added into dichloromethane (12 ml), then trifluoroacetic acid (4 ml) was added, and the reaction was stirred at room temperature for 1 h. The reaction was directly concentrated under reduced pressure to obtain crude compound 41g, which was directly used in the next step.

[0426] LC-MS (ESI): m / z = 326.3 [M+H] + .

[0427] Step 6: Fluorenylmethoxycarbonyl-L-glutamic acid 1-tert-butyl ester (2.94 g, 1.25 mmol) was dissolved in N,N-dimethylformamide (10 ml), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.71 g, 1.88 mmol), N,N-diisopropylethylamine (0.32 g, 2.50 mmol) and the crude compound 41g (1.25 mmol) from the previous step were added, and the reaction was stirred at room temperature for 3 h. After the reaction was completed, ethyl acetate (40 ml) was added for dilution, and water (10 ml x 3) was added for extraction. The organic phase was collected and concentrated, and then compound 41h (0.82 g, 89%) was obtained by rapid separation and purification.

[0428] LCMS m / z = 733.4 [M+H] +

[0429] Step 7: Compound 41h (0.82 g, 1.12 mmol) was dissolved in dichloromethane, and methyl iodide (1.27 g, 8.95 mmol) was added. The reaction was stirred at room temperature overnight, and LCMS was used to monitor the completion of the reaction. Compound 41i was directly obtained by concentration and used in the next step.

[0430] LCMS m / z = 747.4 [M] +

[0431] Step 8: Compound 41i was placed in a 50 ml round-bottom flask, dichloromethane (9 ml) and trifluoroacetic acid (3 ml) were added, and the reaction was stirred at room temperature for 1 h. LCMS was used to monitor the completion of the reaction, and then intermediate 41 (0.68 g, 88%) was obtained by reverse phase purification.

[0432] LCMS m / z = 691.4 [M] +

[0433] Intermediate 42:

[0434] First Step: Take compound 5c (20.0 g, 56.6 mmol) and palladium on carbon (5 g, 10%) as raw materials, and refer to the operation method of the fourth step of Intermediate 35 to obtain compound 42a (13.5 g, 91.2%).

[0435] LC-MS (ESI): m / z = 263.2 [M+H] + .

[0436] Second Step: Take compound 42a (2.5 g, 9.5 mmol) and fluorenylmethoxycarbonyl-L- glutamic acid 1-tert-butyl ester (4.0 g, 9.5 mmol) as raw materials, and refer to the operation method of the first step of Intermediate 14 to obtain compound 42b (2.5 g, 38.8%).

[0437] LC-MS (ESI): m / z = 670.3 [M+H] + .

[0438] Third Step: Take compound 42b (2.5 g, 3.7 mmol) as raw material, and refer to the operation method of the sixth step of Intermediate 35 to obtain the crude product of compound 42c (2.5 g), which is directly used in the next step.

[0439] LC-MS (ESI): m / z = 514.3 [M+H] + .

[0440] Fourth Step: Take compound 42c (2.5 g, 3.7 mmol) and 2-acetyl-5,5-dimethyl-1,3- cyclohexanedione (1.2 g, 6.6 mmol) as raw materials, and refer to the operation method of the third step of Intermediate 38 to obtain Intermediate 42 (1.2 g, 48.0%).

[0441] LC-MS (ESI): m / z = 678.3 [M+H] + .

[0442] Intermediate 43:

[0443] First Step: Add compound 43a (10 g, 62.68 mmol) to diethylene glycol (100 mL), add DIEA (16.17 g, 125.54 mmol), stir at room temperature for 16 h, add water (500 mL) after the reaction is completed, extract with ethyl acetate three times (100 mL x 3), and purify by silica gel chromatography column (PE:EA = 1:1) to obtain compound 43b (9.5 g, 66.1%).

[0444] LC-MS (ESI): m / z = 230.1 [M+H] + .

[0445] Second Step: Take compound 43b (9.5 g, 41.45 mmol) as the raw material, and refer to the operation method of the third step of Intermediate 36 to obtain compound 43c (12 g, 94.2%).

[0446] LC-MS (ESI): m / z = 308.1 [M+H] + .

[0447] Third Step: Take compound 43c (12 g, 39.05 mmol) as the raw material, and refer to the operation method of the fourth step of Intermediate 36 to obtain compound 43d (10 g, 99.3%).

[0448] LC-MS (ESI): m / z = 255.1 [M+H] + .

[0449] Fourth Step: Compound 43d (7.2 g, 28.32 mmol) was dissolved in a mixed solvent of ethanol (72 mL) and water (7.2 mL), and iron powder (7.9 g, 141.6 mmol) and ammonium chloride (7.6 g, 141.6 mmol) were added. Stirring was carried out at 60°C for 4h. After the reaction was completed, filtration and concentration were carried out to obtain the crude product of compound 43e, which was directly used in the next step reaction.

[0450] LC-MS (ESI): m / z = 199.1 [M+H] + .

[0451] Fifth Step: Take compound 43e (5.8 g, crude product) as the raw material, and refer to the operation method of the sixth step of Intermediate 36 to obtain compound 43f (5 g, two-step yield 48.7%).

[0452] LC-MS (ESI): m / z = 363.2 [M+H] + .

[0453] Sixth Step: Take compound 43f (4 g, 11.04 mmol) as the raw material, and refer to the operation method of the eighth step of Intermediate 36 to obtain compound 43g (4.5 g, 53.0%).

[0454] LC-MS (ESI): m / z = 770.3 [M+H] + .

[0455] Seventh Step: Take compound 43g (4.5 g, 5.8 mmol) as the raw material, and refer to the operation method of the ninth step of Intermediate 36 to obtain Intermediate 43 (4 g, 95.9%).

[0456] LC-MS (ESI): m / z = 714.3 [M+H] + .

[0457] Intermediates 44 and 45:

[0458] First step: Compound 44a (20 g, 65.93 mmol) and N,N'-carbonyldiimidazole (13.36 g, 82.41 mmol) were added to tetrahydrofuran (200 ml), stirred for 10 min, reduced to 0 °C, and a sodium borohydride (3.74 g, 98.89 mmol) aqueous solution (60 ml) was added dropwise, and the reaction was carried out at room temperature for 1 h. An aqueous solution of ammonium chloride (100 ml) was added, and ethyl acetate (200 ml x 2) was extracted. The organic phase was combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure, and then purified by silica gel column chromatography to obtain 44b (16 g, 83.37%).

[0459] LC-MS (ESI): m / z = 290.2 [M+H] + .

[0460] Second step: 44b (16 g, 55.29 mmol) was added to dry DMF (150 ml), and sodium hydride (6.64 g, 165.87 mmol, 60%) was added portionwise at 0 °C, and stirred for 20 min. 44c (24.2 g, 55.29 mmol) (synthesis method according to patent WO2017089890A1) was added, and the reaction was carried out at room temperature for 3 h. The reaction solution was poured into water (800 ml), and extracted with ethyl acetate (200 ml). The aqueous phase was adjusted to pH = 4-5 with 1 mol / L hydrochloric acid solution, and the aqueous phase was extracted with ethyl acetate (150 ml x 4). The ethyl acetate was washed with saturated sodium chloride solution, concentrated under reduced pressure, and then purified by chromatography to obtain 44d (7.4 g, 26.84%).

[0461] LC-MS (ESI): m / z = 499.3 [M+H] + .

[0462] Third step: 44d (7 g, 14.04 mmol) was added to methanol (100 ml), and 10% Pd / C (1.5 g) was added, and the reaction was carried out at room temperature for 16 h. The reaction solution was filtered and concentrated under reduced pressure to obtain 44e (5 g, 97.72%).

[0463] LC-MS (ESI): m / z = 365.2 [M+H] + .

[0464] Fourth Step: 44e (5 g, 13.72 mmol), 2-(1-hydroxyethylidene)-5,5-dimethylcyclohexane-1,3-dione (5 g, 27.44 mmol) and triethylamine (2.77 g, 27.44 mmol) were added into ethanol (50 ml) and the reaction was carried out at room temperature for 5 h. The reaction was directly concentrated under reduced pressure and then purified by column chromatography on silica gel to give 44f (5.2 g, 71.7%).

[0465] LC-MS (ESI): m / z = 529.3 [M+H] + .

[0466] Fifth Step: 44f (5 g, 9.46 mmol) was added into dichloromethane (60 ml), and then trifluoroacetic acid (20 ml) was added. The reaction was carried out at room temperature for 1 h. The reaction was directly concentrated under reduced pressure to give 44g (8 g) as a crude product.

[0467] LC-MS (ESI): m / z = 429.3 [M+H] + .

[0468] Sixth Step: 44g (8 g) as a crude product was added into tetrahydrofuran (40 ml), and then 9-fluorenylmethyl-N-succinimidyl carbonate (4.79 g, 14.19 mmol) and saturated aqueous sodium bicarbonate solution (40 ml) were added. The reaction was carried out at room temperature for 5 h. The reaction was extracted with ethyl acetate (50 ml x 3), and the organic phase was washed with saturated sodium chloride solution. The filtrate was concentrated under reduced pressure and then purified by column chromatography. The intermediate 44 (1 g, retention time: 1.678 min) and the intermediate 45 (950 mg, retention time: 1.866 min) were obtained by chiral separation. Analysis conditions: Instrument name: CAS-05-ANA-SFC-D; Column: AS column; Mobile phase: A for CO2; B for 0.05% MNH3 in ethanol; Flow rate: 3 mL / min; Column temperature: 35 °C; Absorption wavelength: 220 nm

[0469] Intermediate 44: LC-MS (ESI): m / z = 651.4 [M+H] + .

[0470] Intermediate 45: LC-MS (ESI): m / z = 651.4 [M+H] + .

[0471] Example 1:

[0472] First Step: The synthesis of 1B was carried out using standard Fmoc chemistry:

[0473] To the reactor was added Rink Amide MBHA Resin resin (1 mmol, 1.4 g, sub: 0.7 mmol / g) and dichloromethane solvent, swelled for 30 min, 20% piperidine / DMF was added, mixed for 30 min.

[0474] Drained and washed with DMF five times.

[0475] Fmoc-Sar-OH protected amino acid solution was added, mixed for 30 seconds before coupling reagent was added, nitrogen bubbled for 1.5 hours, reaction monitored by ninhydrin.

[0476] Drained and washed with DMF three times.

[0477] 20% piperidine / DMF was added, mixed for 30 min.

[0478] Drained and washed with DMF five times.

[0479] Fmoc protected amino acid solution was added, mixed for 30 seconds before coupling reagent was added, nitrogen bubbled for 1.5 hours, reaction monitored by ninhydrin.

[0480] Drained and washed with DMF three times

[0481] Next amino acid coupling steps 5-8 were repeated.

[0482] Final step MeOH wash twice, DCM wash once, MeOH wash twice, vacuum drained to give peptide resin IB (6 g), used directly in next reaction.

[0483] Second step: To a 100 ml reaction flask was added cleavage solution 50 ml (91% trifluoroacetic acid + 4% triisopropylsilane + 3% 1.2-ethanedithiol + 2% water), after stirring well peptide resin IB (6 g) was added, stirred at room temperature for 2 hours. The resin was filtered to give a filtrate, the filtrate was added to 300 ml methyl tert-butyl ether (methyl tert-butyl ether was previously cooled to 0 °C), white flocculent precipitate was formed, centrifuged (3 min at 3000 rpm). The white precipitate was washed with methyl tert-butyl ether three times, vacuum dried to give crude peptide IC as a white solid (2 g), used directly in next reaction.

[0484] Third step: in 2L reaction bottle, add water (750ml), acetonitrile (250ml), 1C (2g, 1.02mmol) in turn, after stirring, slowly drop iodine / acetonitrile solution (0.1mol / L) to the reaction liquid is light yellow, add ascorbic acid to quench, prepare HPLC purification. Separation method: 1. Instrument: waters2767 preparative liquid; chromatographic column: SunFire@Prep C18 (19mm x 250mm). 2. The sample is filtered with 0.45μm filter head to prepare sample solution. 3. Preparation chromatographic conditions: a mobile phase A, B composition: A: 0.1% trifluoroacetic acid / H2O, B: CH3CN; b. Gradient elution: mobile phase content 5%-45%; c. Flow rate: 12ml / min; d. Elution time: 30min, retention time: 16min. Freeze-drying to obtain compound 1 (200mg, purity 99%).

[0485] LCMS m / z = 865.1 [M / 3+H] + .

[0486] Example 2:

[0487] The above material table is used as raw material, and compound 2 (160mg, purity 99%) is synthesized by the method of compound 1.

[0488] LCMS m / z = 846.4 [M / 3+H] + .

[0489] Example 3:

[0490] The above material table is used as raw material, and compound 3 (400mg, purity 99%) is synthesized by the method of compound 1.

[0491] LCMS m / z = 827.9 [M / 3+H] + .

[0492] Example 4:

[0493] The above material table is used as raw material, and compound 4 (80mg, purity 97%) is synthesized by the method of compound 1 after solid phase synthesis to step 14, using 2% hydrazine hydrate / DMF to remove Dde, and then using the method of compound 1 to continue synthesis.

[0494] LCMS m / z = 879.2 [M / 3+H]+ , 659.5 [M / 4 + H] + .

[0495] Example 5:

[0496] Using the above material list as raw material, compound 5 (149 mg, purity 98%) was synthesized by the method of compound 4.

[0497] LCMS m / z = 860.5 [M / 3 + H] + , 645.5 [M / 4 + H] + .

[0498] Example 6:

[0499] Using the above material list as raw material, compound 6 (148 mg, purity 94%) was synthesized by the method of compound 4.

[0500] LCMS m / z = 841.8 [M / 3 + H] + , 631.5 [M / 4 + H] + .

[0501] Example 7:

[0502] First step-third step: using the above material list as raw material, compound 7D (400 mg, purity 99%) was synthesized by the method of compound 1.

[0503] LCMS m / z = 981.4 [M / 2 + H] + , 654.7 [M / 3 + H] + .

[0504] Fourth step: In 10 ml centrifuge tube, 7D (250 mg, 0.127 mmol), succinimidyl- decaglycol-succinimidyl ester (95.6 mg, 0.127 mmol), DMF (3 ml), after stirring, add N, N-diisopropyl ethylamine (65 mg, 0.508 mmol), react at room temperature for 16 h, after LCMS shows the reaction is completed, purify by preparative HPLC. Separation method: 1. Instrument: waters 2767 preparative liquid; column: SunFire@Prep C18 (19 mm x 250 mm). 2. Filter the sample with 0.45 μm filter head to make sample solution. 3. Preparative chromatography conditions: a mobile phase A, B composition: A: 0.1% trifluoroacetic acid / H2O, B: CH3CN; b. Gradient elution: mobile phase content 5%-45%; c. Flow rate: 12 ml / min; d. Elution time: 30 min, retention time: 20 min. Freeze-drying to obtain compound 7 (87 mg, purity 98%).

[0505] LCMS m / z = 1482.9 [M / 3+H] + , 1112.5 [M / 4+H] + , 890.1 [M / 5+H] + .

[0506] Example 15:

[0507] Using the above material table as raw material, compound 15 (140 mg, purity 99%) was synthesized by the method of compound 4.

[0508] LCMS m / z = 1283.2 [M / 2+H] + , 855.8 [M / 3+H] + .

[0509] Example 16:

[0510] Using the above material table as raw material, compound 16 (134 mg, purity 98%) was synthesized by the method of compound 4.

[0511] LCMS m / z = 1311.2 [M / 2+H] + , 874.5 [M / 3+H] + .

[0512] Example 17:

[0513] Using the above material table as raw material, compound 17 (200 mg, purity 99%) was synthesized by the method of compound 4.

[0514] LCMS m / z = 1339.3 [M / 2+H] + 893.3 [M / 3+H] + .

[0515] Example 35:

[0516] First step: synthesis of 35B using standard Fmoc chemistry:

[0517] 1. To the reactor was added 35A (2 mmol, 2 g, sub: 1 mmol / g) and dichloromethane, the resin was swelled for 5 min, then Fmoc-Sar-OH (3 mmol, 0.93 g), N,N- diisopropylethylamine (6 mmol, 1 ml) were added successively, the reaction was carried out at room temperature for 1 h, then methanol (5 ml) was added and the reaction was carried out for 30 min, the solvent was pumped off and the resin was washed with DMF for 3 times.

[0518] 2. 20% piperidine / DMF was added and mixed for 30 min.

[0519] 3. Pumped off and washed with DMF for 5 times.

[0520] 4. Fmoc-protected amino acid solution was added, after mixing for 30 s, coupling reagent was added, nitrogen was bubbled for 1.5 h, and the reaction was monitored by ninhydrin.

[0521] 5. Pumped off and washed with DMF for 3 times

[0522] 6. Steps 2-5 were repeated for the next amino acid coupling.

[0523] 7. The last step was washed with MeOH for 2 times, DCM for 1 time, MeOH for 2 times, vacuum pumped off to obtain the peptide resin 35B (6.5 g), which was directly used for the next step reaction.

[0524] Second step: 30% trifluoroethanol / dichloromethane solution (70 ml) was added to 35B (6.5 g), the reaction was carried out at room temperature for 1 h, then filtered, the filtrate was concentrated under reduced pressure to obtain 35C (2.1 g).

[0525] Third step: In a 50ml flask, add 35C (1.76g, 0.54mmol), intermediate 13 (0.48g, 0.54mmol) in dichloromethane (35ml), stir until clear, then add 1-hydroxybenzotriazole (81mg, 0.6mmol), EDCI (114mg, 0.6mmol), DIPEA (139mg, 1.08mmol) and stir at room temperature for 16h. LCMS shows the starting material is consumed completely. The organic phase is washed with water once, saturated brine once, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 35D (2.0g).

[0526] Fourth step: In a 50ml flask, add cleavage solution 20ml (91% trifluoroacetic acid + 4% triisopropylsilane + 3% 1.2-ethanedithiol + 2% water), stir until clear, then add 35D (2.0g, 0.48mmol) and stir at room temperature for 2h. Filter the resin to give the filtrate, which is added to 300ml methyl tert-butyl ether (methyl tert-butyl ether is cooled to 0°C in advance), and white flocculent precipitates are formed. Centrifuge (3min at 3000rpm). The white precipitate is washed with methyl tert-butyl ether three times, and dried under vacuum to give a white solid crude peptide 35E (1.5g), which is used directly in the next step.

[0527] Fifth step: In a 1L flask, add water (350ml), acetonitrile (150ml), 35E (1.5g, 0.36mmol), stir until clear, then slowly add iodine / acetonitrile solution (0.1mol / L) until the reaction solution is light yellow, and add ascorbic acid to quench. Purify by preparative HPLC. Separation method: 1. Instrument: waters 2767 preparative liquid chromatograph; chromatographic column: SunFire@Prep C18 (19mm x 250mm). 2. Filter the sample with a 0.45μm filter to give a sample solution. 3. Preparative chromatography conditions: a. Mobile phase A and B composition: A: 0.1% trifluoroacetic acid / H2O, B: CH3CN; b. Gradient elution: mobile phase content 5%-45%; c. Flow rate: 12ml / min; d. Elution time: 30min, retention time: 18min. Lyophilize to give compound 35.

[0528] LCMS m / z = 888.5 [M / 3+H] + , 666.7 [M / 4+H] + .

[0529] Example 36:

[0530] Using the above material table as raw material, compound 36 (40mg, 97% purity) is synthesized by the method of compound 4.

[0531] LCMS m / z = 888.2 [M / 3+H] + .

[0532] Example 37:

[0533] Using the above material as raw material, the compound 1 method was used for solid phase synthesis to the 14th step, then DCM (100 mL), phenylsilane (1.08 g, 10 eq), tetrakis (triphenylphosphine) palladium (350 mg, 0.3 eq) was added in turn, and then the compound 1 method was used for continuous synthesis to obtain compound 37 (45 mg, 98%).

[0534] LCMS m / z = 893.0 [M / 3+H] + , 1338.7 [M / 2+H] + .

[0535] Example 38:

[0536] Using the above material as raw material, the compound 4 method was used for synthesis to obtain compound 38 (100 mg, purity 98.7%).

[0537] LCMS m / z = 892.5 [M / 3+H] + .

[0538] Example 39:

[0539] Using the above material as raw material, the compound 4 method was used for synthesis to obtain compound 39 (100 mg, purity 98.5%).

[0540] LCMS m / z = 896.6 [M / 3+H] + .

[0541] Example 40:

[0542] Using the above material as raw material, the compound 8 method was used for synthesis to obtain compound 40 (100 mg, purity 98%).

[0543] LCMS m / z = 976.6 [M / 3+H] + .

[0544] Example 41:

[0545] Using the above material as raw material, compound 41 (0.3 g, 93.3% purity) was synthesized by the method of compound 4.

[0546] LCMS m / z = 897.5 [M / 3+H] + .

[0547] Example 42:

[0548] Using the above material as raw material, compound 42 (70 mg, 94.28%) was synthesized by the method of compound 4.

[0549] LCMS m / z = 896.9 [M / 3+H] + , 1344.7 [M / 2+H] + .

[0550] Example 43

[0551] Using the above material as raw material, compound 43 (150 mg, 99.87% purity) was synthesized by the method of compound 4.

[0552] LCMS m / z = 905.4 [M / 3+H] + .

[0553] Example 44

[0554] Using the above material as raw material, compound 44 (220 mg, 97.76% purity) was synthesized by the method of compound 4.

[0555] LCMS m / z = 905.4 [M / 3+H] + .

[0556] Example 45:

[0557] Using the above material as raw material, compound 45 (60 mg, 94.66%) was synthesized by the method of compound 4.

[0558] LCMS m / z = 914.2 [M / 3+H] + , 1370.9 [M / 2+H] + .

[0559] Example 46:

[0560] Using the above material as raw material, compound 46 (70 mg, 95.11%) was synthesized by the method of compound 4.

[0561] LCMS m / z = 909.5 [M / 3+H] + , 1363.8 [M / 2+H] + .

[0562] Example 47

[0563] Using the above material as raw material, compound 47 (140 mg, purity 96.78%) was synthesized by the method of compound 4.

[0564] LCMS m / z = 926.2 [M / 3+H] + .

[0565] Example 48

[0566] Using the above material as raw material, compound 48 (14 mg, purity 95.87%) was synthesized by the method of compound 4.

[0567] LCMS m / z = 918.4 [M / 3] + .

[0568] Example 49

[0569] Using the above material as raw material, compound 49 (150 mg, purity 97.66%) was synthesized by the method of compound 4.

[0570] LCMS m / z = 913.6 [M / 3+H] + .

[0571] Example 50

[0572] Using the above material as raw material, compound 50 (150 mg, purity 97.66%) was synthesized by the method of compound 4.

[0573] LCMS m / z = 917.6 [M / 3+H] + .

[0574] Biological test evaluation

[0575] The present application is further described in connection with the test examples, but these examples are not meant to limit the scope of the present application.

[0576] 1. IL-23a / IL-12b & IL-23R binding test experiment

[0577] The compound was tested for inhibition of IL-23a / IL-12b & IL-23R binding by TR-FRET method. Protein IL-23a / IL-12b (ACRO, Cat# ILB-H52W5) and IL-23R (ACRO, Cat# ILR-H82F3) solutions were prepared in reaction buffer PPI (PerkinElmer, Cat# 61DB10RDF). The final concentration of IL-23a / IL-12b and IL-23R in the reaction mixture was 0.3 nM. 0.1 μL of the compound to be tested diluted in DMSO was delivered by a nanoliter pipetting system (Echo 655) into a 384-well reaction plate (Grenier, Cat# 784075) and centrifuged at 1000 rpm for 1 minute; 2.5 μL of IL-23a / IL-12b solution was transferred to the 384-well reaction plate and centrifuged at 1000 rpm for 1 minute and incubated at 25°C for 60 minutes; 2.5 μL of IL-23R solution was transferred to the 384-well reaction plate and centrifuged at 1000 rpm for 1 minute; 5 μL of Streptavidin-Tb cryptate (PerkinElmer, Cat# 610SATLA) and Anti6HIS-d2 (PerkinElmer, Cat# 61HISDLB) detection mixture was transferred to the 384-well reaction plate and centrifuged at 1000 rpm for 1 minute and incubated at 25°C for 60 minutes; finally, the HTRF signal (Ratio 665 / 620 nm) was read by a BMG high throughput screening multifunctional microplate reader. The IC50 (half maximal inhibitory concentration) value of the compound was obtained using the four-parameter nonlinear fitting formula (Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))) in the GraphPad Prism software.

[0578] Table 1

[0579] Conclusion: The compound of the present application, such as the compound of the example, has significant inhibitory ability on the binding of IL-23a / IL-12b and IL-23R.

[0580] 2. IL-23R reporter gene assay

[0581] The purpose of this assay is to evaluate the ability of compounds to inhibit the binding of IL23p19 and IL23R in a reporter gene system. HEK-blue IL23 reporter cell line (Invivogen, hkb-il23) is cultured in DMEM + 10% FBS + 100 ug / mL Normocin media. When the density of the cells reaches 80-90%, they are plated at 5000 cells / well in a 384-well plate and incubated overnight at 37°C and 5% CO2. Compound stock solutions are then diluted in DMSO and 40 nL of the dilutions are transferred to the 384-well culture plate by Echo. Incubate for 0.5 hours at 37°C and 5% CO2. Add 40 nL / well of rhIL23 (R&D, 1290-IL) to the 384-well cell culture plate at a final concentration of 1 ng / mL and incubate for 24 hours at 37°C and 5% CO2. Add 18 pL Quanti-Blue TM solution to a new 384-well plate and transfer 2 pL / well of cell culture supernatant to the 384-well plate prepared in Step 6 and incubate for 1 hour at 37°C and 5% CO2. Read the absorbance at 620-655 nM on a BMG. Evaluate the binding ability of the compounds using the following formula and fit IC 50 .

[0582] Inhibition rate calculation formula:

[0583] DMSO control group average

[0584] No IL23 stimulation group average

[0585] IC 50 The calculation formula is as follows: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))

[0586] X: Compound concentration

[0587] Y: Compound inhibition rate

[0588] Conclusion: The compounds of the present application, such as the example compounds, have significant inhibitory ability on the binding of IL23p19 and IL23R.

[0589] 3. IL-23 stimulated pSTAT3 detection assay in PBMC

[0590] Freeze stored human PBMCs were thawed and seeded into plates pre-coated with CD3 antibody at 1x10^6 cells per well. CD28 antibody was then added to the plates and the cells were incubated at 37°C, 5% CO2 for 5 days. On the fifth day, cells were seeded into 96-well plates at a density of 100K cells per well after 4h FBS-starvation stimulation. Diluted compounds were transferred to the 96-well cell culture plates and incubated at 37°C, 5% CO2 for 1h. rhIL23 (R&D, 1290-IL) was added to the cell culture plates and incubated at 37°C, 5% CO2 for 30min. Cells in the wells were lysed with lysis buffer containing 1x PHOSstop solution on ice for 30min and centrifuged at 1000rpm for 1min. The supernatant was then transferred to 96-well ELISA plates and pSTAT3 ELISA assay was performed according to the kit (CST, 7300CA) instructions. Absorbance values at 450nM were read on PHERAstar FSX (BMG LRBTECH). The inhibition rate of the compounds was evaluated using the following formula and IC50 was fitted using Graphpad 50 .

[0591] Inhibition rate calculation formula:

[0592] DMSO control group average

[0593] No IL23 stimulation group average

[0594] IC 50 The calculation formula is as follows: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC50-X)*HillSlope))

[0595] X: Compound concentration

[0596] Y: Compound inhibition rate.

[0597] Conclusion: The compounds of the present application, such as the example compounds, have obvious inhibitory effect on STAT3 phosphorylation.

[0598] 4. Mouse pharmacokinetic test

[0599] 4.1 Test animals: Male Balb / c mice, 20-25g, 6 per compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0600] 4.2 Test design: On the test day, the Balb / c mice were randomly divided into groups according to body weight. Fasting for 12-14h without water restriction 1 day before administration, and feeding 4h after administration.

[0601] Table 2. Dosing information

[0602] Note: Intravenous administration vehicle: PBS; Intragastric administration vehicle: PBS

[0603] Before and after administration, 0.06 mL of blood was taken from the orbit under isoflurane anesthesia, placed in an EDTA K2 centrifuge tube, centrifuged at 5000 rpm at 4°C for 10 min, and the plasma was collected. The blood sampling time points for the intravenous group and the intragastric group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24 h. Before analysis, all samples were stored at -80°C, and the samples were quantitatively analyzed by LC-MS / MS.

[0604] Table 3. Pharmacokinetic parameters of test compounds in mouse plasma

[0605] Conclusion: The compounds of the present application, such as the compounds of the examples, have good pharmacokinetic characteristics in mice.

[0606] 5. Rat pharmacokinetic test

[0607] 5.1 Test animals: Male SD rats, about 220 g, 6-8 weeks old, 6 rats per compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0608] 5.2 Test design: On the test day, the SD rats were randomly divided by weight. Fasting for 12-14 h without water restriction 1 day before administration, and feeding 4 h after administration. Administration: 6 rats per group (6 rats per test compound), 3 intravenous administration (intravenous group) + 3 oral gavage administration (gavage group); intravenous administration vehicle: NS or PBS; intragastric administration vehicle: NS or PBS.

[0609] Before and after administration, 0.15 mL of blood was taken from the orbit under isoflurane anesthesia, placed in an EDTA K2 centrifuge tube, centrifuged at 5000 rpm at 4°C for 10 min, and the plasma was collected. The blood sampling time points for the intravenous group and the intragastric group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 24, 48, 72 h. Brain tissue was taken at 24 h after administration, the surface residual blood was washed with cold physiological saline, and the homogenate was treated after being absorbed. Before analysis, all samples were stored at -80°C, and the samples were quantitatively analyzed by LC-MS / MS.

[0610] Table 4. Pharmacokinetic parameters of test compounds in rat plasma

[0611] -: not applicable. Compounds 35, 37, and 39 were sampled up to 24 h.

[0612] Conclusion: The compound of the present application, such as the compound of the example, has good rat in vivo pharmacokinetic characteristics, showing good exposure AUC, low clearance rate, and long half-life.

[0613] 6. Beagle pharmacokinetic test

[0614] 6.1 Test animals: male beagles, about 8-11 kg, 6 per compound, purchased from Beijing Mass Biotechnology Co., Ltd.

[0615] 6.2 Test method: On the test day, the beagles were randomly grouped according to body weight. Fasting but not water restriction for 12-14 h before administration, and feeding 4 h after administration.

[0616] 1 mL of blood was taken through the jugular vein or the limbs before and after administration and placed in an EDTA K2 centrifuge tube. Centrifugation at 5000 rpm for 10 min at 4°C, and collection of plasma. The blood sampling time points for the intravenous group and the gavage group were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 120, 144 h. Before analysis, all samples were stored at -80°C, and LC-MS / MS was used for quantitative analysis of the samples.

[0617] Conclusion: The compound of the present application, such as the compound of the example, has good dog in vivo pharmacokinetic characteristics.

[0618] 7. Monkey pharmacokinetic test

[0619] 7.1 Test animals: male cynomolgus monkeys, 3-5 kg, 3-6 years old, 4 per compound. Purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0620] 7.2 Test method: On the test day, the monkeys were randomly grouped according to body weight. Fasting but not water restriction for 14-18 h before administration, and feeding 4 h after administration.

[0621] 1.0 mL of blood was taken through the limbs before and after administration and placed in an EDTA K2 centrifuge tube. Centrifugation at 5000 rpm for 10 min at 4°C, and collection of plasma. The blood sampling time points for the intravenous group were: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, 72, 96, 120, 144 h. Before analysis, all samples were stored at -80°C, and LC-MS / MS was used for quantitative analysis of the samples.

[0622] Conclusion: The compound of the present application, such as the compound of the example, has good monkey in vivo pharmacokinetic characteristics.

[0623] 8. Plasma stability test

[0624] The plasma samples of human, monkey, dog, rat and mouse were used to evaluate the plasma stability of the compound.

[0625] The plasma samples with the concentration of 1000 ng / mL were prepared and divided into EP tubes for time points of 0 h and 6 h; the 0 h sample was directly added with the internal standard acetonitrile solution, and the 6 h sample was placed at 37℃ for the corresponding time and then added with the internal standard acetonitrile solution. The LC-MS / MS method was used to detect the concentration of the test substance in the sample, and the residual rate was calculated by the peak area ratio of the test substance to the internal standard in the time point sample and the zero time sample.

[0626] 9. Gastrointestinal fluid stability test

[0627] 9.1. Solution preparation

[0628] Dilute hydrochloric acid: take hydrochloric acid 23.4 ml, dilute with water to 1000 ml, and you get it.

[0629] Artificial gastric juice preparation: take dilute hydrochloric acid 1.64 ml, add water about 80 ml and pepsin 1 g, shake well, and dilute with water to 100 ml, and you get it.

[0630] Prepare 0.1 mol / L NaOH: weigh NaOH 0.4 g, dissolve in 100 ml water.

[0631] Preparation of artificial intestinal juice: take potassium dihydrogen phosphate 0.68 g, add water 50 ml to dissolve, adjust the pH value to 6.8 with 0.1 mol / L sodium hydroxide solution; take another trypsin 1 g, add water to dissolve, mix the two liquids, and dilute with water to 1000 ml, and you get it.

[0632] 9.2. Sample preparation

[0633] Take about 12.5 mg of sample into a 25 ml capacity bottle, add artificial gastric juice (or artificial intestinal juice) to dissolve and dilute to the calibration line.

[0634] 9.3. Analysis method

[0635] Instrument model Agilent 1260 Infinity; mobile phase A: 10 mmol / L K2HPO4 mobile phase B: acetonitrile; column: phenomenex Gemini @ 3um C18 150*4.6mm; wavelength: 224nm; column temperature: 30℃; sample disc temperature: 37℃; injection time: 35min; injection volume: 10ul; injection mode: gradient injection; gradient method.

[0636] 9.4. Sample detection and results

[0637] Sampling detection: First, the blank solution (artificial gastric juice or artificial intestinal juice) is detected, then the prepared sample (freshly prepared) is placed into the sample plate and immediately injected, and then each sample is injected.

[0638] Conclusion: The compounds of the present application, such as the compounds of the examples, have good artificial gastric and intestinal fluid stability.

[0639] 10. hERG potassium ion channel effect test

[0640] Experimental platform: electrophysiology manual patch clamp system

[0641] Cell line: Chinese hamster ovary (CHO) cell line stably expressing hERG potassium ion channel

[0642] Experimental method: CHO (Chinese Hamster Ovary) cells stably expressing hERG potassium channel are used to record hERG potassium channel current at room temperature by whole-cell patch clamp technique. Glass microelectrode is drawn by glass electrode embryo (BF150-86-10, Sutter) through a drawing instrument, and the tip resistance of the electrode after being filled with internal solution is about 2-5 MΩ. The glass microelectrode is inserted into the amplifier probe and connected to the patch clamp amplifier. The clamping voltage and data recording are controlled and recorded by pClamp 10 software through computer, the sampling frequency is 10 kHz, and the filter frequency is 2 kHz. After obtaining the whole-cell recording, the cell is clamped at -80 mV, and the step voltage for inducing hERG potassium current (I hERG) is given from -80 mV to +20 mV for 2 s, and then repolarized to -50 mV for 1 s, and then returned to -80 mV. This voltage stimulation is given every 10 s, and after the hERG potassium current is determined to be stable (at least 1 minute), the drug administration process is started. Each test concentration of the compound is given for at least 1 minute, and at least 2 cells are tested for each concentration (n≥2).

[0643] Data processing: pClamp 10, GraphPad Prism 5 and Excel software are used for data analysis and processing. The inhibition degree of different compound concentrations on hERG potassium current (hERG tail current peak value induced at -50 mV) is calculated by the following formula:

[0644] Inhibition% = [1-(I / Io)]x100%

[0645] Wherein, Inhibition% represents the inhibition percentage of the compound on hERG potassium current, and I and Io represent the amplitude of hERG potassium current after and before drug administration, respectively.

[0646] The IC50 of the compound is calculated by fitting the following equation using GraphPad Prism 5 software:

[0647] Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X)*HillSlope))

[0648] Where X is the Log value of the test article concentration, Y is the percent inhibition at the corresponding concentration, Bottom and Top are the minimum and maximum percent inhibition, respectively.

[0649] Conclusion: The compounds of the application, e.g. the example compounds, do not inhibit hERG.

[0650] 11. CYP enzyme inhibition test

[0651] The purpose of this study is to evaluate the effects of test articles on the activities of five isoforms of human liver microsomal cytochrome P450 (CYP) (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) using in vitro test systems. Specific probe substrates of CYP450 isoforms are incubated with human liver microsomes and different concentrations of test articles, and the reaction is initiated by adding reduced nicotinamide adenine dinucleotide phosphate (NADPH). After the reaction is completed, the samples are processed and the specific metabolites produced from the substrates are quantitatively determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) to measure the changes in CYP enzyme activities, calculate IC50 values, and evaluate the inhibitory potential of test articles on each CYP enzyme subtype. The incubation concentration is 0-30 μM under the test conditions.

[0652] Conclusion: The compounds of the application, e.g. the example compounds, do not inhibit CYP enzymes.

[0653] 12. Liver microsomal stability test

[0654] In this experiment, human, canine, rat and mouse liver microsomes of five species are used as in vitro models to evaluate the metabolic stability of test articles.

[0655] At 37°C, 1 μM of test article is incubated with microsomal protein and coenzyme NADPH, and the reaction is terminated by adding ice-cold acetonitrile containing an internal standard at a certain time (5, 10, 20, 30, 60 min). The concentration of the test article in the sample is detected by LC-MS / MS method, and the T1 / 2 is calculated from the ln value of the residual rate of the drug in the incubation system and the incubation time, and the liver microsomal intrinsic clearance CLint(mic) and the liver intrinsic clearance CLint(Liver) are further calculated.

[0656] Conclusion: The compounds of the application, e.g. the example compounds, have good liver microsomal stability.

[0657] 13. Caco2 permeability test

[0658] The test uses single layer Caco-2 cells, incubated in triplicate in 96-well Transwell plates. Transport buffer solution (HBSS, 10 mM HEPES, pH 7.4 ± 0.05) containing the compound of the application (2 μM) or control compounds digoxin (10 μM), nadolol (2 μM) and metoprolol (2 μM) is added to the donor wells on the apical side or the basolateral side. DMSO-containing transport buffer solution is added to the corresponding receiver wells. After incubation at 37 ± 1 °C for 2 hours, the cell plates are removed and an aliquot of the sample is taken from the apical and basolateral sides into new 96-well plates. Subsequently, acetonitrile containing internal standard is added to precipitate the proteins. The samples are analyzed using LC MS / MS and the concentrations of the compound of the application and the control compounds are determined. The concentration data are used to calculate the apparent permeability coefficients for transport from the apical side to the basolateral side and vice versa, and thus the efflux ratio. The integrity of the monolayer after 2 hours of incubation is evaluated using the leakage of fluorescein.

[0659] Conclusion: The compounds of the application, for example the example compounds, have good efflux ratio indicators.

[0660] 14. Pharmacodynamic test of the test substance in the IL-23-induced rat skin inflammation model

[0661] Model establishment: On Day -1, the animals are randomly divided into groups according to ear thickness and body weight. The sham control group is injected intradermally with 1 × PBS in the left ear from Day 0 to Day 4, once a day. The model control group is injected intradermally with IL-23 in the left ear from Day 0 to Day 4, once a day. The dosing group is injected intradermally with IL-23 in the left ear from Day 0 to Day 4, once a day. The animals in all groups are subjected to modeling and dosing after the first daily test compound administration.

[0662] Dosing of the test compound: The test compound is administered by gavage QD from Day 0 to Day 4; the sham control group and the model control group are given the corresponding volume of vehicle.

[0663] Data collection and analysis: The data are collected using Excel software. The data are analyzed using Prism 10.1.2 (Graph pad software, Inc.) software.

[0664] Ear thickness inhibition rate (%) = (ear thickness of dosed animals - average ear thickness of the sham control group) / (average ear thickness of the model control group - average ear thickness of the sham control group) × 100%

[0665] Results: The compounds of the present application, such as the compounds of the examples, can significantly inhibit the ear thickness in the rat IL-23-induced skin inflammation model. The Model group of rats was injected with IL-23 in the left ear, and the Sham group was injected with PBS in the ear. Oral administration of the compounds can significantly inhibit the ear thickness under QD administration.

[0666] 15. PD study of test drugs in a rat model induced by imiquimod

[0667] IL-17A is the main downstream cytokine of IL-23. In this experiment, IL-17A was used as a PD index to study the inhibitory effect of test drugs on IL-17A in an oral administration rat model.

[0668] SD rats (purchased from Vivotan) were randomly divided into 3 groups according to body weight and back skin thickness. From Day 0, the rats were sensitized by applying imiquimod (Sichuan Mingxin Pharmaceutical Co., Ltd.) on their back skin every afternoon for 4 days. From Day 2, the rats were administered test drugs by gavage once a day for 7 days. The detailed administration scheme is shown in Table 5. The last administration was performed in the morning of Day 4. Whole blood was collected at 0h, 1h, 4h, 8h, 24h, and 48h after administration, and anticoagulated with heparin sodium.

[0669] The collected whole blood was diluted 1:4 with RPMI-1640, and then transferred to a 96-well plate (240uL). After incubation at 37℃ for 30 minutes, IL-23 at a final concentration of 40ng / mL and IL-1β at a final concentration of 40ng / mL were used for stimulation for 24 hours. The supernatant was then collected and the content of IL-17A in the supernatant was detected by Elisa method. Graphpad Prism was used for plotting, and the average IL-17A concentration of each animal in the Vehicle group was used as the Control group. The relative content of the average IL-17A concentration of the compound at each time point corresponding to the Control group was calculated as follows:

[0670] IL-17A relative content % = average IL-17A concentration of the administration group / average IL-17A concentration of the Control group * 100%

[0671] Table 5 Grouping information table

[0672] Data analysis: two-way ANOVA analysis was used, and method was used to compare the significant analysis of IL-17A secretion levels of the two drugs at the same time point.

[0673] According to the results of the PD study, the compounds of the present application, such as the compounds of the examples, can effectively inhibit the secretion of IL-17A.

Claims

A cyclic peptide compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of Formula (I), (II): Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Glu-Asn-[3-Pal]-Xa 13 (I) (SEQ ID NO: 1) Xa1-His-Thr-Xa4-Xa5-[Pen]-Xa7-[2-Nal]-Xa9-Xa 10 -Asn-[3-Pal]-Xa 13 (II) (SEQ ID NO: 2) wherein: Xa1 is Pen or (D)Pen; Xa4 is Trp(7-methyl) or Trp(7-cyclopropyl); Xa5 is Gin or Lys; Xa7 is Xa9 is Thp or Xa 10 Glu, Xa 13 Sarc or the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group; The cyclic peptide compound is conjugated with a modifying group at at least one of Xa1, Xa5, Xa7, Glu, Xa 10 , Xa 13 , 3-Pal or Thr, the modifying group being -L-R, L being -L1-L2-L3-, wherein L3 is attached to R and L1 is a bond, L2 comprises a PEG linker, L3 is a spacer unit, and R is: n is 4-15; with the proviso that: (1) when Xa4 is Trp (7-cyclopropyl), Xa5 is Gin, Xa9 is and only when the modifying group is conjugated at Xa1, the modifying group is not: (2) when Xa4 is Trp (7-cyclopropyl), Xa5 is Lys, Xa9 is at the time, and only when the modifying group is conjugated at Xa5, the modifying group is not: The cyclic peptide compound according to claim 1, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, the L is -L1-L2-L3-, wherein: L1 is a bond; L2 is p is 0-50, q is 1-50, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is L3is a bond, in combination, m is 0-5, preferably L3 is The cyclic peptide compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, characterized in that, Xa1 is Pen; Xa4 is Trp(7-methyl); Xa5 is Lys; Xa7 is Xa9 is Thp. The cyclic peptide compound according to claim 3, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, characterized in that, Xa1 is Pen; Xa4 is Trp(7-methyl); Xa5 is Lys; Xa7 is Xa9 is Thp; Xa 10 is Glu; Xa 13 Sarc; the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group; said cyclic peptide compound is conjugated to a modifying group at at least one of Xa1, Xa5, Xa7, Glu, Xa 13 3-Pal or Thr. The cyclic peptide compound according to any one of claims 1 to 4, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, The L is: p is 1-5, q is 1-2, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is Preferably L is: p is 1-5, q is 1-2, X is -(4-6 membered heterocycloalkyl)-, -(4-6 membered heterocycloalkyl)-O-, -NH-(5-6 membered heteroaryl)-O-, preferably X is More preferably L is: X is The cyclic peptide compound according to claim 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, The R is: n is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15; preferably R is: The cyclic peptide compound according to claim 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, characterized in that, the cyclic peptide compound is conjugated with a modifying group at Glu. A cyclic peptide compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein the cyclic peptide compound has an amino acid sequence of Formula (III), [Pen]-His-Thr-Trp(7-Me)-Lys(Ac)-[Pen]-Xa7-[2-Nal]-THP-Xa 10 -Asn-[3-Pal]-Sarc(III) (SEQ ID NO: 3) Xa7 is Xa 10 To the cyclic peptide compound is cyclized by a Pen-Pen disulfide bond, and the cyclic peptide compound is optionally linked to a protecting group. A peptide dimer compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein two cyclic peptide compounds having an amino acid sequence of Formula (I), (II) as claimed in claim 1 are linked via one or more linkers selected from diethylene glycol, iminodiacetic acid, beta-Ala-iminodiacetic acid or polyethylene glycol to form a peptide dimer. The compound according to any one of claims 1 to 9, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or characterized in that, wherein the compound is selected from one of the structures of Table I. A pharmaceutical composition comprising a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1-10, and a pharmaceutically acceptable carrier and / or excipient. The pharmaceutical composition according to claim 11, comprising 1-1500 mg of the compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and / or excipient. Use of a compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1-10 or a pharmaceutical composition as claimed in claim 11 or 12 in the manufacture of a medicament for the prevention and treatment of a disease or disorder in a subject in which IL-23 is overexpressed in a diseased tissue. The use according to claim 13, wherein the disease or disorder in which IL-23 is overexpressed comprises inflammatory bowel disease, Crohn's disease and psoriasis. A method for treating a disease in a mammal or a human, said method comprising administering to the subject a therapeutically effective amount, preferably 1-1500 mg, of a compound according to any one of claims 1-10, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, said disease preferably inflammatory bowel disease, Crohn's disease and psoriasis.

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