Polypeptides for repairing skin or mucosal damage and uses thereof

Abstract

The present application relates to a polypeptide for repairing skin injury or mucosa injury, which has obvious effects of reducing pathological development of acute and chronic gastrointestinal diseases, promoting repair of skin or mucosa injury, treating tumors and the like, and use of the polypeptide for repairing skin injury or mucosa injury. Specifically, the present application relates to a polypeptide capable of binding to FGFR1 receptor, which plays a role in inhibiting or promoting cell proliferation, vascular proliferation, collagen formation and the like by blocking or activating FGF / FGFR1 signal transmission.

Description

[0001] This application is a divisional application of Chinese invention patent application number CN202111650580.5, filed on December 30, 2021, entitled "A type of polypeptide for repairing skin or mucous membrane damage and its application". Technical Field

[0002] This invention relates to polypeptides for repairing skin or mucosal damage, which have significant effects in alleviating the pathological progression of acute and chronic gastrointestinal diseases, promoting the repair of skin or mucosal damage, and treating tumors; and the use of said polypeptides for repairing skin trauma or mucosal damage. Specifically, this invention relates to polypeptides capable of binding to the FGFR1 receptor, which exert their effects of inhibiting or promoting cell proliferation, angiogenesis, and collagen formation by blocking or activating FGF / FGFR1 signaling. Background Technology

[0003] At least 19 members of the fibroblast growth factor (FGF) family have been identified, playing crucial roles in development, angiogenesis, and wound healing. Their functions are achieved through binding to fibroblast growth factor receptors (FGFRs), which, via signal transduction systems, activate corresponding genes. Fibroblast growth factor receptors are a class of transmembrane tyrosine kinase receptors, and four types have been identified (FGFR1, FGFR2, FGFR3, and FGFR4), each encoded by a separate gene. Their functions are similar, involving the regulation of cell proliferation, differentiation, migration, and some pathological processes. FGFRs are widely distributed on the surface of target cells, such as epithelial cells, fibroblasts, and vascular endothelial cells. Phenotypic alterations of FGFRs are associated with hereditary diseases and malignant transformation of cells.

[0004] Fibroblast growth factors (FGFs) initiate intracellular signal transduction by binding to receptors (FGFRs), regulating cell proliferation, differentiation, and migration. Activation of FGFRs plays an important role in angiogenesis, embryonic development, tumor growth, and wound repair.

[0005] Skin and / or mucous membrane injury is a common pathological feature of many diseases. Skin injury refers to damage to normal skin (tissue) caused by external traumatic factors such as surgery, external force, heat, electric current, chemicals, and low temperature, as well as internal factors such as local blood supply disorders. It is often accompanied by the disruption of skin integrity and the loss of a certain amount of normal tissue, while also impairing normal skin function. It is also called a wound or trauma. Currently, protein / peptide drugs such as basic fibroblast growth factor, epidermal growth factor, platelet-derived growth factor, granulocyte-macrophage colony-stimulating factor, and growth hormone have significant effects on wound repair, skin care, anti-wrinkle, and anti-aging. However, the long amino acid sequences of these protein / peptide drugs lead to high preparation costs and poor stability, thus limiting their application.

[0006] Chronic gastritis is a chronic inflammation of the gastric mucosa, a common and frequently occurring disease in gastroenterology. Clinically, chronic gastritis refers to chronic inflammation of the gastric mucosa (pathologically manifested as mononuclear cell and lymphocyte infiltration) and / or glandular atrophic lesions caused by various factors. Chronic gastritis is more common in middle-aged and elderly people. Its onset is related to age but not to gender. It has a slow onset and a protracted course, making it difficult to cure and challenging to treat.

[0007] FGF / FGFR1 signaling is essential for normal cell growth, but excessive FGF secretion or insufficient expression can lead to various diseases. Studies have found that breast cancer, glioma, and liver cancer cells all express high levels of FGFR1, and abnormal FGFR1-mediated signaling is closely related to fibrotic diseases such as pulmonary fibrosis and cirrhosis. Furthermore, under conditions of skin or mucous membrane injury, activation of FGF / FGFR1 signaling can promote cell and tissue repair. Currently, many challenges remain in the treatment of skin and mucous membrane diseases and tumors, requiring further research and solutions from scientists to discover more and better drugs with therapeutic potential.

[0008] Given that current treatments cannot meet clinical needs, there is still a need for peptides that can better treat skin and mucous membrane diseases and tumors, such as acute and chronic gastrointestinal diseases and skin or mucous membrane injuries. Summary of the Invention

[0009] Through extensive experiments and research, the inventors of this invention have discovered that the polypeptide of this invention can bind to the FGFR1 receptor and exert its effects of inhibiting or promoting cell proliferation, angiogenesis, and collagen formation by blocking or activating FGF / FGFR1 signal transduction. Therefore, the polypeptide of this invention has significant effects in alleviating the pathological development of acute and chronic gastrointestinal diseases, promoting the repair of skin or mucous membrane damage, and treating tumors.

[0010] In a first aspect, the present invention provides compounds of formula (I) or physiologically compatible salts thereof, wherein the compounds of formula (I) are as follows:

[0011] H-Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-OH(I),

[0012] in

[0013] Xaa1 is Pro and missing;

[0014] Xaa2 represents Ala, which is missing;

[0015] Xaa3 represents Ser, Ala, Thr, and is missing;

[0016] Xaa4 is Met, Met(O), Val, Ala, Leu, Ile, Thr, Glu, Arg, or is missing;

[0017] Xaa5 is Gln, Ala, Glu, Pro, Lys, or is missing;

[0018] Xaa6 is Ala, Ser, Val, Asp, or missing;

[0019] Xaa7 is Ser, Ala, Thr, Pro, Val, or missing;

[0020] Xaa8 is Leu, Ala, Val, Tyr, or missing;

[0021] Xaa9 is Glu, Gln, Ala, Leu, Asp or missing;

[0022] Xaa10 is Ala, Ser, or missing;

[0023] Xaa11 is Glu, Gln, Ala, Leu, Asp or is missing;

[0024] Xaa12 is Ala, Ser, or missing;

[0025] Xaa13 is Lys, Ala, Arg, His, or missing;

[0026] Xaa14 is Gly, Ala, Pro or missing;

[0027] Xaa15 is Lys, Arg, Ala, His, or missing;

[0028] Xaa16 is Ala, Ser, or missing;

[0029] Xaa17 is Glu, Gln, Asp, Asn, Ala, Lys or is missing;

[0030] Xaa18 is Asp, Glu, Leu, Ala, Gln, or is missing; and

[0031] Xaa19 is Asp, Glu, or missing; and

[0032] The condition is that at least four of Xaa1, Xaa2, Xaa3, Xaa4, Xaa5, Xaa6, Xaa7, Xaa8, Xaa9, Xaa10, Xaa11, Xaa12, Xaa13, Xaa14, Xaa15, Xaa16, Xaa17, Xaa18, and Xaa19 are not missing.

[0033] In one implementation, both Xaa1 and Xaa2 are missing.

[0034] In one embodiment, Xaa3 is Ser or missing, preferably Ser.

[0035] In one embodiment, Xaa4 is Met or Met(O) or missing, preferably Met or Met(O).

[0036] In one embodiment, Xaa3-Xaa4 are Ser-Met, Ser-Met(O), Ser-Val, Ser-Ala, Ser-Leu, Ser-Ile, Thr-Met or missing, preferably Ser-Met, Ser-Met(O) or missing, more preferably Ser-Met.

[0037] In one embodiment, Xaa5 is Gln or missing, preferably Gln.

[0038] In one implementation, Xaa6 is Ala or missing, preferably Ala.

[0039] In one implementation, Xaa7 is Ser or missing, preferably Ser.

[0040] In one embodiment, Xaa8 is Leu or missing, preferably Leu.

[0041] In one embodiment, Xaa5-Xaa6-Xaa7-Xaa8 are Gln-Ala-Ser-Leu, Gln-Ala-Ser-Ala, Ala-Ala-Ser-Leu, Gln-Ala-Ala-Leu, Glu-Ala-Ser-Leu, Gln-Ala-Thr-Leu, Gln-Ala-Ser-Val, Ala-Ser-Thr-Leu, Pro-Val-Pro-Leu, Lys-Asp-Val-Tyr, or are absent, preferably Gln-Ala-Ser-Leu.

[0042] In one implementation, Xaa9 is Glu or Gln.

[0043] In one implementation, Xaa10 is Ala.

[0044] In one implementation, Xaa11 is Glu, Gln, or Ala.

[0045] In one implementation, Xaa12 is Ala.

[0046] In one implementation, Xaa13 is Lys.

[0047] In one embodiment, Xaa9-Xaa10-Xaa11-Xaa12 are Glu-Ala-Glu-Ala, Gln-Ala-Gln-Ala, Ala-Ala-Glu-Ala, Glu-Ala-Ala-Ala, Leu-Ala-Glu-Ala, Gl u-Ala-Leu-Ala, Gln-Ala-Glu-Ala, Glu-Ala-Gln-Ala, Asp-Ala-Glu-Ala, Glu-Ala-Asp-Ala, Glu-Ser-Glu-Ala, Glu-Ala-Glu-Ser or missing.

[0048] In one embodiment, Xaa9-Xaa10-Xaa11-Xaa12-Xaa13 is Glu-Ala-Glu-Ala-Lys, Gln-Ala-Gln-Ala-Lys, Glu-Ala-Glu-Ala-A la,Ala-Ala-Glu-Ala-Lys,Glu-Ala-Ala-Ala-Lys,Glu-Ala-Glu-Ala-Arg,Glu-Ala-Glu-Ala-His,Leu-Ala-Glu- Ala-Lys, Glu-Ala-Leu-Ala-Lys, Gln-Ala-Glu-Ala-Lys, Glu-Ala-Gln-Ala-Lys, Asp-Ala-Glu-Ala-Lys, Glu-Ala -Asp-Ala-Lys, Glu-Ser-Glu-Ala-Lys, Glu-Ala-Glu-Ser-Lys, Glu-Ala-Glu-Ala or deletion, preferably Glu-Ala-Glu-Ala-Lys.

[0049] In one implementation, Xaa14 is Gly.

[0050] In one implementation, Xaa15 is Lys.

[0051] In one embodiment, Xaa14-Xaa15 is Gly-Lys, Gly-Arg, Gly-Ala, Gly-His, Ala-Lys, Pro-Lys, or absent, preferably Gly-Lys.

[0052] In one implementation, Xaa16 is Ala.

[0053] In one implementation, Xaa17 is Glu.

[0054] In one implementation, Xaa16-Xaa17 are Ala-Glu, Ala-Gln, Ala-Asp, Ala-Asn, Ala-Ala, Ala-Lys, Ser-Glu, or are absent.

[0055] In one implementation, Xaa18-Xaa19 are Asp, Glu, Leu, Ala, Gln-Asp, Gln-Glu, or are absent.

[0056] In one embodiment, the compound is selected from:

[0057] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 1);

[0058] Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 2);

[0059] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys (compound 3);

[0060] Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys (compound 4);

[0061] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys (compound 5);

[0062] Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys (compound 6);

[0063] Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 7);

[0064] Ser-Met-Gln-Ala-Ser-Leu (compound 8);

[0065] Gln-Ala-Ser-Leu (compound 9);

[0066] Gly-Lys-Ala-Glu (compound 10);

[0067] Ser-Met(O)-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 11);

[0068] Ser-Val-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 12);

[0069] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln (compound 13);

[0070] Ser-Met-Gln-Ala-Ser-Leu-Gln-Ala-Gln-Ala-Lys-Gly-Lys-Ala-Glu (compound 14);

[0071] Ser-Met-Gln-Ala-Ser-Ala-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 15);

[0072] Ser-Ala-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 16);

[0073] Ser-Leu-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 17);

[0074] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Ala-Gly-Lys-Ala-Glu (Compound 18);

[0075] Ser-Met-Gln-Ala-Ser-Leu-Ala-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 19);

[0076] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Ala-Ala-Lys-Gly-Lys-Ala-Glu (Compound 20);

[0077] Ser-Met-Ala-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 21);

[0078] Ser-Ile-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 22);

[0079] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Asp (Compound 23);

[0080] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Asn (Compound 24);<>

[0081] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Arg-Ala-Glu (Compound 25);

[0082] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Ala-Ala-Glu (Compound 26);

[0083] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Ala (Compound 27);

[0084] Ser-Met-Gln-Ala-Ala-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 28);

[0085] Ala-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 29);

[0086] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Arg-Gly-Lys-Ala-Glu (Compound 30);

[0087] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-His-Ala-Glu (Compound 31);

[0088] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-His-Gly-Lys-Ala-Glu (Compound 32);

[0089] Ser-Met-Gln-Ala-Ser-Leu-Leu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 33);

[0090] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Leu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 34);

[0091] Ser-Met-Gln-Ala-Ser-Leu-Gln-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 35);

[0092] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Gln-Ala-Lys-Gly-Lys-Ala-Glu (Compound 36);

[0093] Ser-Met-Glu-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 37);

[0094] Ser-Met-Gln-Ala-Ser-Leu-Asp-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 38);

[0095] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Asp-Ala-Lys-Gly-Lys-Ala-Glu (Compound 39);

[0096] Thr-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 40);

[0097] Ser-Met-Gln-Ala-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 41);

[0098] Ser-Met-Gln-Ala-Ser-Val-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 42);

[0099] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ser-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 43);

[0100] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ser-Lys-Gly-Lys-Ala-Glu (Compound 44);

[0101] Glu-Ala-Glu-Ala (Compound 45);

[0102] Glu-Ala-Glu-Ala-Lys-Gly-Lys (Compound 46);

[0103] Thr-Ala-Ser-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 47);

[0104] Ser-Thr-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 48);

[0105] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Asp (Compound 49);

[0106] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Glu (Compound 50);

[0107] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu-Leu (Compound 51);

[0108] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu-Ala (Compound 52);

[0109] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Lys-Gln-Asp(Compound 53);

[0110] Ala-Glu-Pro-Val-Pro-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (Compound 54);

[0111] Pro-Ala-Ala-Glu-Pro-Val-Pro-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu(Compound 55);

[0112] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Ala-Lys-Ala-Glu (compound 56);

[0113] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Pro-Lys-Ala-Glu (compound 57);

[0114] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Lys-Gln-Glu (compound 58);

[0115] Arg-Lys-Asp-Val-Tyr-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (compound 59);

[0116] Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ser-Glu (compound 60); or

[0117] Glu-Ala-Glu-Ala-Ala-Gly-Lys-Ala-Glu (Compound 61).

[0118] In a second aspect, the present invention provides the following compounds or physiologically compatible salts thereof:

[0119] Phe-Asp-Ala-Leu-Lys-Gln-Gln-Phe-Gln-Ala-Phe-Gln-Leu-Glu (compound 62);

[0120] His-Cys-Leu-Ala-Gly-Leu-Lys-Lys-Asp-Leu-Glu-Asp-Leu-Glu (compound 63);

[0121] Glu-Ile-Asn-Gln-Leu-Glu-Leu-Ile-Lys-Gln-Ala-Ser-Ile (compound 64);

[0122] Ala-Ala-Arg-Leu-Ala-Asp-Glu-Leu-Arg-Ala-Glu (compound 65);

[0123] Glu-Thr-Leu-Gln-Arg-Lys-Asn-Lys-Glu (compound 66);

[0124] Val-Asp-Ala-Ala-Val-Leu-Glu-Lys-Leu-Glu (compound 67);

[0125] Ala-Ala-Val-Leu-Asp-Lys-Leu-Glu (compound 68);

[0126] Ala-Ala-Val-Leu-Glu-Lys-Leu-Glu (compound 69);

[0127] Lys-Phe-Tyr-Ser-Gln-Ser-Thr-Ala-Ser-Ser-Ser-Tyr-Ala-Tyr-Pro-Ser-His-Phe-Gly-Pro-Ala-Gly-Phe-Ser-Gly-Ser-His-Ser (compound 70);

[0128] Val-Asp-Ala-Ala-Val-Ile-Glu-Lys-Ile-Glu (compound 71);

[0129] Val-Asp-Ala-Ala-Met-Val-Leu-Leu-Thr-Arg (compound 72); or

[0130] Val-Asp-Ala-Ala-Val-Leu-Met-Leu-Arg-Thr (compound 73).

[0131] For convenience, the H on the left and the OH on the right are omitted when describing the compounds of the present invention in this application.

[0132] Thirdly, the present invention provides a method for repairing skin trauma, the method comprising contacting the skin trauma with a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof; or, the present invention provides the use of a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament or cosmetic for repairing skin trauma; or the present invention provides a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for repairing skin trauma. In one embodiment, the skin trauma is related to, but is not limited to, epidermal inflammation, mechanical and surgical wounds, burns and scalds, ulcers, fistulas, pressure sores, and skin damage caused by radiotherapy and chemotherapy. In one embodiment, the skin trauma refers to damage to normal skin caused by external injury factors such as surgery, external force, heat, electric current, chemicals, low temperature, and internal factors such as local blood supply disorders. In one embodiment, the skin trauma is often accompanied by disruption of skin integrity and loss of a certain amount of normal tissue. In another embodiment, the skin trauma includes impairment of normal skin function.

[0133] This invention provides a method for promoting HaCAT cell proliferation, the method comprising contacting the cells with a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof. Alternatively, this invention provides the use of a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for promoting HaCAT cell proliferation; or this invention provides a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for promoting HaCAT cell proliferation.

[0134] Fourthly, the present invention provides a method for repairing mucosal damage, the method comprising administering a compound of the present invention or a physiologically compatible salt thereof to a subject or contacting the mucosal damage with the compound of the present invention or a physiologically compatible salt thereof. Alternatively, the present invention provides the use of the compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for repairing mucosal damage; or the present invention provides the compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for repairing mucosal damage.

[0135] In one embodiment, the mucosal injury is mucosal injury within cavities such as the digestive or respiratory systems.

[0136] Mucosal damage to the digestive system is associated with oral, esophageal, and gastrointestinal diseases. Oral diseases include oral ulcers, stomatitis, gingivitis, periodontitis, etc.; esophageal diseases include esophagitis, esophageal ulcers, etc.; and gastrointestinal diseases include, but are not limited to, mucosal damage caused by radiotherapy and chemotherapy, such as chronic gastritis, chronic atrophic gastritis, acute gastritis, gastroduodenal ulcers, functional gastrointestinal disorders, dyspepsia, precancerous lesions, digestive system tumors, gastrointestinal bleeding, gastroesophageal reflux disease, acute and chronic enteritis, ulcerative colitis, Crohn's disease, and radiotherapy and chemotherapy.

[0137] In a preferred embodiment, the digestive tract mucosa includes the gastric mucosa and the intestinal mucosa. In a preferred embodiment, the mucosal injury is gastric mucosal injury caused by irritants or drugs, or stress. Irritants include hydrochloric acid, ethanol, or alcohol, and drugs include nonsteroidal anti-inflammatory drugs such as aspirin or indomethacin.

[0138] This invention provides a method for preventing, alleviating, or treating gastrointestinal diseases or eliminating inflammatory edema, the method comprising administering to a subject a compound of the invention or a physiologically compatible salt thereof. Alternatively, this invention provides the use of a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for preventing, alleviating, or treating gastrointestinal diseases or eliminating inflammatory edema; or this invention provides a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for preventing, alleviating, or treating gastrointestinal diseases or eliminating inflammatory edema. The digestive tract diseases mentioned include those related to the oral cavity, esophagus, and gastrointestinal tract. Oral diseases include oral ulcers, stomatitis, gingivitis, periodontitis, etc.; esophageal diseases include esophagitis, esophageal ulcers, etc.; and gastrointestinal diseases include chronic gastritis, chronic atrophic gastritis, acute gastritis, gastroduodenal ulcers, functional gastrointestinal disorders, dyspepsia, precancerous lesions, digestive system tumors, gastrointestinal bleeding, gastroesophageal reflux disease, acute and chronic enteritis, ulcerative colitis, Crohn's disease, and mucosal damage caused by radiotherapy and chemotherapy; but are not limited thereto. In one embodiment, the prevention, mitigation, or treatment of the digestive tract diseases is achieved by regulating stem cell proliferation and differentiation. The method utilizes the compounds of the present invention or their physiologically compatible salts to protect or repair damage to the gastrointestinal mucosa, such as the gastric or intestinal mucosa, thereby preventing, mitigating, or treating gastrointestinal diseases.

[0139] This invention provides a method for repairing skin trauma or mucosal damage, the method comprising administering to a subject a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof. Alternatively, this invention provides the use of a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament or cosmetic for repairing skin trauma or mucosal damage; or this invention provides a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for repairing skin trauma or mucosal damage.

[0140] This invention provides a method for blocking or activating FGF / FGFR1 signaling, the method comprising administering to a subject a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof, causing it to bind to an FGFR1 receptor, thereby blocking or activating FGF / FGFR1 signaling and exerting an inhibitory or promotion effect on cell proliferation, angiogenesis, and collagen formation. Alternatively, this invention provides the use of the compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for blocking or activating FGF / FGFR1 signaling; or this invention provides the compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof, which is used to block or activate FGF / FGFR1 signaling. In a further embodiment, the compound of the invention (i.e., the compound of the first or second aspect above) binds to an FGFR1 receptor, thereby blocking or activating FGF / FGFR1 signaling and exerting an inhibitory or promotion effect on cell proliferation, angiogenesis, and collagen formation. In one embodiment, the present invention provides a method for activating FGF / FGFR1 signaling, the method comprising administering to a subject a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof, causing it to bind to an FGFR1 receptor, thereby activating FGF / FGFR1 signaling and exerting an inhibitory or promoting effect on cell proliferation, angiogenesis, and collagen formation; or, the present invention provides a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for activating FGF / FGFR1 signaling; or the present invention provides the use of a compound of the present invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for activating FGF / FGFR1 signaling.

[0141] This invention provides a method for treating diseases related to FGFR targets, the method comprising administering to a subject a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof. Alternatively, this invention provides the use of a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof in the preparation of a medicament for treating diseases related to FGFR targets; or this invention provides a compound of the invention (i.e., the compound of the first or second aspect above) or a physiologically compatible salt thereof for treating diseases related to FGFR targets. In one embodiment, the FGFR target is FGFR1. In one embodiment, the disease related to the FGFR target is cancer or tumor.

[0142] In the methods of the present invention described above, the compounds of the present invention or their physiologically compatible salts are administered by oral, injection, subcutaneous, or other means.

[0143] Fifthly, the present invention provides a pharmaceutical, food, health product, cosmetic, or daily product composition, said composition comprising the compound of the present invention or a physiologically compatible salt thereof, and a physiologically acceptable carrier. In one embodiment, the physiologically acceptable carrier comprises a pharmaceutically acceptable carrier or a cosmetically acceptable carrier. The pharmaceutical, food, health product, cosmetic, or daily product composition can be prepared according to conventional pharmaceutical or cosmetic techniques, including mixing the compound of the present invention as an active ingredient with a carrier and preparing it into the desired dosage form according to conventional techniques. The composition of the present invention can be formulated into oral administration formulations, mucosal administration formulations, injectable formulations, inhaled formulations, and topical formulations as needed.

[0144] The polypeptides of this invention are not homologous to known polypeptides, facilitating the synthesis of high-purity polypeptides. Furthermore, the polypeptides of this invention can bind to the FGFR1 receptor to exert biological effects, exhibiting significant therapeutic effects on skin and mucous membrane diseases caused by various factors. These polypeptides can be used to treat acute and chronic gastrointestinal diseases, tumors, and related diseases. The polypeptides of this invention have a small molecular weight and, when administered orally, significantly reduce inflammation and edema, promote the repair of gastrointestinal mucosal damage, and alleviate the pathological development of acute and chronic gastritis and peptic ulcers. They also promote skin wound repair, shorten wound healing time, and regulate immune function. Oral administration is also effective. In addition, the polypeptides of this invention remain effective even after disinfection with iodine preparations or hydrogen peroxide when applied to skin wounds, whereas epidermal growth factor, when applied to skin wounds, will have its structure destroyed after disinfection with iodine preparations or hydrogen peroxide and will not function. Attached Figure Description

[0145] The accompanying drawings are provided for the purpose of further illustrating this application and are not intended to limit the scope of this application.

[0146] Figure 1 The mass spectrum of compound 62 is shown.

[0147] Figure 2 The mass spectrum of compound 63 is shown.

[0148] Figure 3 The mass spectrum of compound 1 is shown.

[0149] Figure 4 The mass spectrum of compound 64 is shown.

[0150] Figure 5 The mass spectrum of compound 65 is shown.

[0151] Figure 6 The mass spectrum of compound 66 is shown.

[0152] Figure 7 The mass spectrum of compound 67 is shown.

[0153] Figure 8 The mass spectrum of compound 68 is shown.

[0154] Figure 9 The mass spectrum of compound 69 is shown.

[0155] Figure 10 The mass spectrum of compound 70 is shown.

[0156] Figure 11 The mass spectrum of compound 71 is shown.

[0157] Figure 12 The mass spectrum of compound 72 is shown.

[0158] Figure 13 The mass spectrum of compound 73 is shown.

[0159] Figure 14 A schematic diagram of the solid-phase synthesis steps of peptides is shown.

[0160] Figure 15 The therapeutic effect of compound 1 on a mouse model of chronic atrophic gastritis was demonstrated.

[0161] Figure 16 The study demonstrated the proliferative effect of compound 53 on HaCaT cells.

[0162] Figure 17 The study demonstrated the proliferative effect of compound 1 on HaCaT cells.

[0163] Figure 18 The study demonstrated the proliferative effect of compound 64 on HaCaT cells.

[0164] Figure 19The study demonstrated the proliferative effect of compound 65 on HaCaT cells.

[0165] Figure 20 The study demonstrated the proliferative effect of compound 66 on HaCaT cells.

[0166] Figure 21 The study demonstrated the proliferative effect of compound 57 on HaCaT cells.

[0167] Figure 22 The study demonstrated the proliferative effect of compound 68 on HaCaT cells.

[0168] Figure 23 The study demonstrated the proliferative effect of compound 69 on HaCaT cells.

[0169] Figure 24 The proliferative effect of compound 70 on HaCaT cells was demonstrated.

[0170] Figure 25 The proliferative effect of compound 71 on HaCaT cells was demonstrated.

[0171] Figure 26 The study demonstrated the proliferative effect of compound 72 on HaCaT cells.

[0172] Figure 27 The study demonstrated the proliferative effect of compound 73 on HaCaT cells.

[0173] Figure 28 The proliferative effect of compound 1 on HMEC-1 cells was demonstrated.

[0174] Figure 29 The proliferative effect of compound 64 on HMEC-1 cells was demonstrated.

[0175] Figure 30 The proliferative effect of compound 71 on HMEC-1 cells was demonstrated.

[0176] Figure 31 The proliferative effect of compound 72 on HMEC-1 cells was demonstrated.

[0177] Figure 32 The proliferative effect of compound 73 on HMEC-1 cells was demonstrated.

[0178] Figure 33 The proliferative effect of compound 63 on RSC96 cells was demonstrated.

[0179] Figure 34 The proliferative effect of compound 1 on RSC96 cells was demonstrated.

[0180] Figure 35The proliferation-promoting effect of compound 65 on RSC96 cells was demonstrated.

[0181] Figure 36 The proliferative effect of compound 67 on RSC96 cells was demonstrated.

[0182] Figure 37 The effect of compound 68 on the proliferation of RSC96 cells was shown.

[0183] Figure 38 Compound 70 was shown to inhibit the proliferation of RSC96 cells.

[0184] Figure 39 The effect of compound 71 on the proliferation of RSC96 cells was shown.

[0185] Figure 40 The proliferative effect of polypeptide compound 73 on RSC96 cells was demonstrated. Detailed Implementation

[0186] The term "physiologically compatible salt" refers to a salt form that is physiologically compatible (i.e., pharmacologically acceptable) and substantially non-toxic to the individual to whom the compounds of the present invention will be administered. Physiologically compatible salts of the compounds of the present invention include conventional and stoichiometric acid addition salts or base addition salts formed from suitable, non-toxic organic or inorganic acids or inorganic bases.

[0187] The term "subject" refers to an animal, preferably a mammal, and most preferably a human. Specifically, the term "subject" refers to a mammal or human with skin trauma and / or mucosal damage. Those skilled in the art will understand that the repair of skin trauma and / or mucosal damage of the present invention can be applied for cosmetic (i.e., non-therapeutic) and therapeutic purposes. Therefore, the term "skin damage" in this application also includes skin damage repaired for cosmetic purposes, such as wrinkles (e.g., wrinkles caused by ultraviolet radiation), skin lines, cracks, lumps, large pores (e.g., associated with appendage structures such as sweat ducts, sebaceous glands, or hair follicles), or unevenness or roughness, loss of skin elasticity (loss and / or inactivation of functional elastin), sagging (including puffiness around the eyes and jaw), loss of skin firmness, loss of skin tightness, loss of the ability to recover after skin deformation, discoloration (including dark circles), freckles, grayish-yellow skin tone, hyperpigmented skin areas such as age spots and freckles, keratin, abnormal differentiation, hyperkeratosis, degeneration of elastic tissue, destruction of collagen, and other tissue changes in the stratum corneum, dermis, epidermis, vascular system of the skin (e.g., telangiectasia or bifurcated vessels), and subcutaneous tissue, especially other tissue changes in the subcutaneous tissue close to the skin.

[0188] The following description of the present invention is based on specific experiments and is not intended to limit the scope of protection of the present invention.

[0189] Example 1: Chemical Synthesis of Peptides

[0190] The peptide compounds were synthesized using conventional solid-phase synthesis methods, involving multiple cycles of resin swelling, amino acid substitution, deprotection, washing, amino acid activation, condensation, washing, and deprotection, followed by final cleavage and side chain deprotection. This application utilizes a peptide synthesizer for synthesis.

[0191] Table 1. English names, abbreviations, and corresponding Chinese names of solvents and reagents, etc.

[0192]

[0193] The following uses compound 1 (Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu) as an example to illustrate the method for the synthesis and purification of compound 1. The method includes steps 1, preparation of fully protected peptide resin; 2, cleavage and deprotection; 3, purification (salt replacement) and lyophilization.

[0194] Step 1: Preparation of fully protected peptide resin

[0195] (1) Preparation of Fmoc-Glu(OtBu)-resin: Weigh 13.48 g of 2-Chlorotrityl Chloride Resin (SD = 0.75 mmol / g) and dichloromethane (DCM) and add them sequentially to the 6 synthesis tubes of the peptide synthesizer. Set the peptide synthesizer to the correct settings.

[0196] The substitution reaction procedure was followed to perform the substitution reaction of the first amino acid and remove the protecting group Fmoc. The specific reaction procedure is as follows: The resin was swollen with DCM for 20 minutes and then drained. In an activation flask, Fmoc-Aa(n)-OH (3 eq, 6 mmol) was dissolved in DMF (40 mL), and then DIPEA (7 eq, 14 mmol) was added. The Fmoc-Aa(n)-OH / DIPEA DMF mixture was added to the reaction flask containing the resin and reacted for 1 hour, then drained. 20% MeOH / DMF (40 mL) was added and reacted for 30 minutes for end-capping. After draining, the mixture was washed 3 times with DMF, and then the protecting group Fmoc was removed with 20% PIP / DMF (v / v), followed by washing 5 times with DMF solution.

[0197] (2) Preparation of fully protected peptide resin: The circulating water bath temperature was adjusted to 35 degrees Celsius. Using the coupling program set in the peptide synthesizer, amino acids were coupled one by one according to the peptide sequence, starting from the (n-1)th amino acid. The specific reaction process is as follows: Fmoc-Aa(n-1)-OH (3 eq, 6 mmol) and HOBt (3 eq, 6 mmol) were dissolved in DMF (40 mL) in an activation flask, and then DIC (4 eq, 8 mmol) was added to activate the amino acids. The DMF mixture of Fmoc-Aa(n-1)-OH / HOBt / DIC was added to the amino acid resin in the reaction flask for coupling. After reacting for 1 hour, the solvent was removed, and the solid resin was washed 3 times with DMF solution. Then, the protecting group Fmoc was removed with 20% PIP / DMF (v / v) and washed 5 times with DMF solution. The above steps were repeated until the entire amino acid sequence was coupled. The amount of each amino acid and condensing agent is shown in Table 2.

[0198] Step 2, Cutting

[0199] Transfer the resin from the peptide synthesizer to a manual synthesis tube with a sieve plate and wash five times with DCM. Add 200 mL of cleavage agent (TFA:TIS:H2O = 95:2.5:2.5, v / v) to the synthesis tube, and then bubble the reaction under nitrogen (N2) for 1.5–3 hours.

[0200] After the cleavage reaction was complete, the cleavage agent was filtered into a 500 mL round-bottom flask. After vacuum concentration to one-quarter of the original cleavage agent volume, 10 times the current volume of methyl tert-butyl ether was added, and the mixture precipitated to obtain a white solid. The resulting mixture was filtered and washed three times with 50 mL of methyl tert-butyl ether. The resulting crude peptide product was then placed in a vacuum drying oven to remove excess solvent until the crude peptide was in powder form. 11.46 g of crude peptide was obtained, with a crude yield of 61.5%.

[0201] Step 3: Purification (salt replacement) freeze-drying

[0202] (1) Purification of peptides (HPLC)

[0203] A. Chromatographic parameters

[0204] Column: Dynamic axial compression column, 80*250 mm; Packing material: Daisogel C18 (SP-100-8-ODS-P)

[0205] Eluent A: 0.1% trifluoroacetic acid aqueous solution (v / v)

[0206] Elution Buffer B: Acetonitrile

[0207] Flow rate: 180 mL / min

[0208] UV detection wavelength: 220 nm

[0209] B. Operating Procedures

[0210] a) Dissolve the crude peptide in water and / or acetonitrile and filter through a 0.45 μm filter membrane; b) Inject the sample; c) Elute with an acetonitrile-water mobile phase gradient; d) Collect the target peptide eluent; e) Concentrate by rotary evaporation.

[0211] (2) Polypeptide salt swapping (HPLC)

[0212] A. Chromatographic parameters

[0213] Column: Dynamic axial compression column, 80*250 mm; Packing material: Daisogel C18 (SP-100-8-ODS-P)

[0214] Eluent A1: 0.1 M acetic acid

[0215] Eluent A2: 0.025 M acetic acid – 0.1 M ammonium acetate

[0216] Elution Buffer B: Acetonitrile

[0217] Flow rate: 180 mL / min

[0218] UV detection wavelength: 220 nm

[0219] B. Operating Procedures

[0220] a) Equilibrate column with 95% A1 + 5% B; b) Injection; c) Equilibrate column with 95% A2 + 5% B; d) Gradient elution with A1 and B; e) Collection of target peptide eluent; f) Rotary evaporation concentration; g) Freeze-drying.

[0221] Table 2. Amounts of amino acids and condensing agents used in the chemical synthesis of peptides.

[0222]

[0223] Other compounds were synthesized in a similar manner to that used for compound 1. Results are shown in Table 3 and other parts of the specification.

[0224] Table 3. Synthesized polypeptide compounds

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234] Note: Double charge peaks indicate that the target molecule has 2 protons, and triple charge peaks indicate that the target molecule has 3 protons; N / A indicates that weighing is difficult and the actual weight is not included.

[0235] Compound 1: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0236] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O25S; m / z: 517.25855 ([M+3H]3+), 775.38204 ([M+2H]2+), 1549.74923 ([M+H]+).

[0237] 1H NMR (600 MHz, D2O+MeOD)δ4.63–4.50 (m, 1H), 4.27 (t, J = 7.3 Hz, 1H), 4.19 (dd, J = 9.3, 5.1 Hz, 1H), 4.14 (t, J = 5.5 Hz, 1H), 4.09 (q, J =7.2 Hz, 2H), 4.05–3.93 (m, 9H), 3.85–3.76 (m, 2H), 3.75–3.62 (m, 4H), 2.75(t, J = 7.6 Hz, 4H), 2.39–2.21 (m, 8H), 2.15 (t, J = 7.5 Hz, 2H), 2.03–1.71(m, 23H, AcOH), 1.69–1.20 (m, 16H), 1.20–1.13 (m, 12H), 0.68 (d, J = 6.3 Hz, 3H), 0.62 (d, J = 6.3 Hz, 3H).

[0238] Compound 2: Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0239] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C55H94N16O22; m / z: 444.56747 ([M+3H]3+), 666.34654 ([M+2H]2+), 1331.68030 ([M+H]+).

[0240] Compound 3: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys

[0241] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C55H96N16O21S; m / z: 450.56546 ([M+3H]3+), 675.34210 ([M+2H]2+), 1349.67108 ([M+H]+).

[0242] Compound 4: Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys

[0243] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C47H82N14O18; m / z: 377.87399 ([M+3H]3+), 566.30565 ([M+2H]2+), 1131.59986 ([M+H]+).

[0244] Compound 5: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys

[0245] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C47H81N13O19S; m / z: 582.78356 ([M+2H]2+), 1164.55736 ([M+H]2+).

[0246] Compound 6: Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys

[0247] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C39H67N11O16; m / z: 473.74506 ([M+2H]2+), 946.48291 ([M+H]2+).

[0248] Compound 7: Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0249] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C38H65N11O16; m / z: 466.73868 ([M+2H]2+), 932.46713 ([M+H]+).

[0250] Compound 8: Ser-Met-Gln-Ala-Ser-Leu

[0251] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C25H45N7O10S; m / z: 636.30254 ([M+H]+).

[0252] Compound 9: Gln-Ala-Ser-Leu (acetate)

[0253] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C17H31N5O7; m / z: 418.22972 ([M+H]+).

[0254] 1H NMR (600 MHz, D2O)δ4.43–4.33 (m, 2H), 4.20–4.15 (m, 1H), 4.00 (t,J = 6.6 Hz, 1H), 3.85–3.76 (m, 2H), 2.46–2.35 (m, 2H), 2.17–2.04 (m, 2H), 2.00–1.96 (m, 2H, AcOH), 1.58–1.51 (m, 3H), 1.37 (d, J = 7.2 Hz, 3H), 0.89–0.76 (m, 6H).

[0255] Compound 10: Gly-Lys-Ala-Glu

[0256] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C16H29N5O7; m / z: 404.21472 ([M+H]+).

[0257] Compound 11: Ser-Met(O)-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0258] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O26S; m / z: 522.58959 ([M+3H]3+), 783.37985 ([M+2H]2+), 1565.74624 ([M+H]+).

[0259] 1H NMR (600 MHz, D2O)δ4.59–4.46 (m, 3H), 4.37 (t, J = 5.6 Hz, 1H), 4.32–4.22 (m, 7H), 4.22–4.17 (m, 3H), 4.16–4.10 (m, 2H), 3.98–3.92 (m, 2H),3.90 (s, 1H), 3.88–3.77 (m, 3H), 2.94 (t, J = 7.5 Hz, 5H), 2.90–2.81 (m, 1H),2.66 (s, 3H), 2.41–2.19 (m, 9H), 2.17–1.67 (m, 23H, AcOH), 1.66–1.53 (m, 7H), 1.48–1.36 (m, 4H), 1.35–1.32 (m, 12H), 0.87 (d, J = 5.6 Hz, 3H), 0.81 (d, J =5.7 Hz, 3H).

[0260] Compound 12: Ser-Val-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0261] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O25; m / z: 506.59990 ([M+3H]3+), 759.39412 ([M+2H]2+), 1517.77125 ([M+H]+).

[0262] 1H NMR (600 MHz, D2O) δ4.37 (t, J = 5.5 Hz, 1H), 4.33–4.23 (m, 7H), 4.22–4.11 (m, 6H), 3.98–3.79 (m, 6H), 2.95 (t, J = 7.5 Hz, 4H), 2.41–2.25 (m,8H), 2.08–2.00 (m, 5H), 1.99–1.68 (m, 15H, AcOH), 1.67–1.54 (m, 7H), 1.50–1.37 (m, 4H), 1.37–1.33 (m, 12H), 0.92–0.86 (m, 9H), 0.82 (d, J = 5.7 Hz, 3H).

[0263] Compound 13: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln (acetate)

[0264] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H109N19O24S; m / z: 516.92919 ([M+3H]3+), 774.88810 ([M+2H]2+), 1548.75961 ([M+H]+).

[0265] 1H NMR (600 MHz, D2O)δ4.47 (dd, J = 8.3, 6.1 Hz, 1H), 4.39–4.34 (m,1H), 4.31–4.16 (m, 10H), 4.16–4.08 (m, 2H), 3.97–3.94 (m, 2H), 3.94–3.79 (m,4H), 2.94 (t, J = 7.6 Hz, 4H), 2.60–2.47 (m, 2H), 2.40–2.22 (m, 8H), 2.10–1.98 (m, 9H), 1.98–1.84 (m, 13H, AcOH), 1.84 –1.67 (m, 4H), 1.67–1.54 (m,7H), 1.48–1.37 (m, 4H), 1.36–1.32 (m, 12H), 0.87 (d, J = 5.8 Hz, 3H), 0.82(d, J = 5.8 Hz, 3H).

[0266] Compound 14: Ser-Met-Gln-Ala-Ser-Leu-Gln-Ala-Gln-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0267] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H110N20O23S; m / z: 516.60130 ([M+3H]3+), 774.39599 ([M+2H]2+), 1547.77410 ([M+H]+).

[0268] 1H NMR (600 MHz, D2O)δ4.69–4.61 (m, 1H), 4.13 (t, J = 7.3 Hz, 1H), 4.09–3.93 (m, 4H), 3.92–3.78 (m, 10H), 3.74–3.65 (m, 2H), 3.64–3.51 (m, 4H),2.98–2.96 (m, 1H), 2.95–2.92 (m, 2H), 2.65–2.59 (m, 4H), 2.28–1.97 (m, 10H),1.89–1.57 (m, 21H, AcOH), 1.57–1.18 (m, 11H), 1.18–1.08 (m, 4H), 1.07–1.00(m, 12H), 0.55 (d, J = 6.4 Hz, 3H), 0.50 (d, J = 6.4 Hz, 3H).

[0269] Compound 15: Ser-Met-Gln-Ala-Ser-Ala-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0270] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C60H102N18O25S; m / z: 503.24185 ([M+3H]3+), 754.35857 ([M+2H]2+), 1507.70677 ([M+H]+).

[0271] Compound 16: Ser-Ala-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0272] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C61H104N18O25; m / z: 497.25701 ([M+3H]3+), 745.38093 ([M+2H]2+), 1489.74746 ([M+H]+).

[0273] Compound 17: Ser-Leu-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0274] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H110N18O25; m / z: 766.40505 ([M+2H]2+).

[0275] Compound 18: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Ala-Gly-Lys-Ala-Glu

[0276] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C60H101N17O25S; m / z: 746.85331 ([M+2H]2+).

[0277] Compound 19: Ser-Met-Gln-Ala-Ser-Leu-Ala-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0278] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C61H106N18O23S; m / z: 497.92179 ([M+3H]3+), 746.37833 ([M+2H]2+), 1491.74166 ([M+H]+).

[0279] Compound 20: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Ala-Ala-Lys-Gly-Lys-Ala-Glu

[0280] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C61H106N18O23S; m / z: 497.92359 ([M+3H]3+), 746.38084 ([M+2H]2+), 1491.74509 ([M+H]+).

[0281] Compound 21: Ser-Met-Ala-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0282] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C61H105N17O24S; m / z: 498.25060 ([M+3H]3+), 746.87141 ([M+2H]2+).

[0283] Compound 22: Ser-Ile-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0284] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H110N18O25; m / z: 511.27255 ([M+3H]3+), 766.40318 ([M+2H]2+), 1531.78999 ([M+H]+)

[0285] Compound 23: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Asp

[0286] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H106N18O25S; m / z: 512.58493 ([M+3H]3+), 768.37218 ([M+2H]2+), 1535.72417 ([M+H]+).

[0287] Compound 24: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Asn

[0288] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H107N19O24S; m / z: 512.25851 ([M+3H]3+), 767.88243 ([M+2H]2+), 1534.34795 ([M+H]+).

[0289] Compound 25: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Arg-Ala-Glu

[0290] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N20O25S; m / z: 526.59167 ([M+3H]3+), 789.38318 ([M+2H]2+), 1577.76111 ([M+H]+).

[0291] Compound 26: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Ala-Ala-Glu

[0292] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C60H101N17O25S; m / z: 746.85430 ([M+2H]2+), 1492.68936 ([M+H]+)

[0293] Compound 27: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Ala

[0294] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C61H106N18O23S; m / z: 497.92090 ([M+3H]3+), 746.37592 ([M+2H]2+), 1491.74463 ([M+H]+).

[0295] Compound 28: Ser-Met-Gln-Ala-Ala-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0296] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O24S; m / z: 511.92444 ([M+3H]3+), 767.38306 ([M+2H]2+), 1533.75671 ([M+H]+).

[0297] Compound 29: Ala-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0298] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O24S; m / z: 511.92638 ([M+3H]3+), 767.38435 ([M+2H]2+), 1533.75280 ([M+H]+).

[0299] 1H NMR (600 MHz, D2O) δ4.38 (dd, J = 8.4, 6.0 Hz, 1H), 4.32 (t, J =5.6 Hz, 1H), 4.26– 4.12 (m, 10H), 4.08 (dd, J = 8.5, 4.9 Hz, 1H), 4.00 (q, J = 7.1 Hz, 1H), 3.90–3.84 (m, 1H), 3.84– 3.73 (m, 3H), 2.89 (t, J = 7.6 Hz, 4H), 2.54–2.42 (m, 2H), 2.37–2.21 (m, 8H), 2.05–1.95 (m, 8H), 1.95–1.49 (m,25H, AcOH), 1.43 (d, J = 7.1 Hz, 3H), 1.41–1.31 (m, 4H), 1.30 (t, J = 7.2 Hz,12H), 0.83 (d, J = 5.8 Hz, 3H), 0.77 (d, J = 6.0 Hz, 3H).

[0300] Compound 30: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Arg-Gly-Lys-Ala-Glu

[0301] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N20O25S; 526.59256 ([M+3H]3+), 789.38355 ([M+2H]2+), 1577.74446 ([M+H]+).

[0302] Compound 31: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-His-Ala-Glu

[0303] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H103N19O25S; m / z: 520.24450 ([M+3H]3+), 779.86149 ([M+2H]2+), 1558.70931 ([M+H]+).

[0304] Compound 32: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-His-Gly-Lys-Ala-Glu

[0305] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H103N19O25S; m / z: 520.24531 ([M+3H]3+), 779.86349 ([M+2H]2+), 1558.71094 ([M+H]+).

[0306] Compound 33: Ser-Met-Gln-Ala-Ser-Leu-Leu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0307] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H112N18O23S; m / z: 767.38324 ([M+2H]2+), 1533.75610 ([M+H]+).

[0308] Compound 34: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Leu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0309] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H112N18O23S; m / z: 767.39618 ([M+2H]2+), 1533.79517 ([M+H]+).

[0310] 1H NMR (600 MHz, D2O) δ4.39 (dd, J = 8.2, 6.2 Hz, 1H), 4.30 (dd, J =9.3, 5.1 Hz, 1H), 4.26 (t, J = 5.4 Hz, 1H), 4.23–4.04 (m, 11H), 3.89 (d, J =4.7 Hz, 2H), 3.87–3.74 (m, 4H), 2.86 (t, J = 7.7 Hz, 4H), 2.50–2.40 (m, 2H), 2.40–2.32 (m, 4H), 2.26 (t, J = 7.5 Hz, 2H), 2.13–2.05 (m, 1H), 2.00–1.96 (m,5H), 1.96–1.83 (m, 13H, AcOH), 1.78–1.60 (m, 4H), 1.60–1.51 (m, 8H), 1.51–1.43 (m, 2H), 1.42–1.29 (m, 4H), 1.29–1.26 (m, 12H), 0.80 (t, J = 5.6 Hz, 6H), 0.74 (d, J = 6.0 Hz, 6H).

[0311] Compound 35: Ser-Met-Gln-Ala-Ser-Leu-Gln-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0312] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H109N19O24S; m / z: 774.88964 ([M+2H]2+), 1548.76276 ([M+H]+).

[0313] Compound 36: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Gln-Ala-Lys-Gly-Lys-Ala-Glu

[0314] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H109N19O24S; m / z: 516.92993 ([M+3H]3+), 774.88916 ([M+2H]2+), 1548.76034 ([M+H]+).

[0315] Compound 37: Ser-Met-Glu-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0316] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H107N17O26S; m / z: 775.87388 ([M+2H]2+), 1550.72354 ([M+H]+).

[0317] Compound 38: Ser-Met-Gln-Ala-Ser-Leu-Asp-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0318] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H106N18O25S; m / z: 768.37273 ([M+2H]2+), 1535.72499 ([M+H]+).

[0319] Compound 39: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Asp-Ala-Lys-Gly-Lys-Ala-Glu

[0320] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H106N18O25S; m / z: 768.37189 ([M+2H]2+), 1535.72715 ([M+H]+).

[0321] Compound 40: Thr-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0322] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H110N18O25S; m / z: 521.92978 ([M+3H]3+), 782.38824 ([M+2H]2+), 1563.75498 ([M+H]+).

[0323] Compound 41: Ser-Met-Gln-Ala-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0324] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H110N18O25S; m / z: 521.92871 ([M+3H]3+), 782.38721 ([M+2H]2+), 1563.76697 ([M+H]+).

[0325] Compound 42: Ser-Met-Gln-Ala-Ser-Val-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0326] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H106N18O25S; m / z: 768.37357 ([M+2H]2+).

[0327] Compound 43: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ser-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0328] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O26S; m / z: 522.58964 ([M+3H]3+), 783.37864 ([M+2H]2+).

[0329] Compound 44: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ser-Lys-Gly-Lys-Ala-Glu

[0330] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O26S; m / z: 783.37737 ([M+2H]2+), 1565.72717 ([M+H]+).

[0331] Compound 45: Glu-Ala-Glu-Ala (acetate)

[0332] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C16H26N4O9; m / z: 419.17728 ([M+H]+).

[0333] 1H NMR (600 MHz, DMSO-d6)δ8.60 (d, J = 7.1 Hz, 1H), 8.27 (d, J = 7.8Hz, 1H), 7.90 (d, J = 6.8 Hz, 1H), 4.29 (p, J = 7.0 Hz, 1H), 4.19 (td, J =8.3, 5.1 Hz, 1H), 4.02 (p, J = 7.1 Hz, 1H), 3.66 (t, J = 6.3 Hz, 1H), 2.31(t, J = 7.6 Hz, 2H), 2.23 (t, J = 7.9 Hz, 2H), 1.95–1.79 (m, 5H, AcOH), 1.79–1.70 (m, 1H), 1.27–1.21 (m, 6H).

[0334] Compound 46: Glu-Ala-Glu-Ala-Lys-Gly-Lys

[0335] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C30H53N9O12; m / z: 732.38880 ([M+H]+).

[0336] Compound 47: Thr-Ala-Ser-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0337] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C58H100N16O24; m / z: 469.24518 ([M+3H]3+), 703.36326 ([M+2H]2+), 1405.71314 ([M+H]+).

[0338] Compound 48: Ser-Thr-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu (acetate)

[0339] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C62H106N18O26; m / z: 507.25973 ([M+3H]3+), 760.38380 ([M+2H]2+), 1519.74995 ([M+H]+).

[0340] 1H NMR (600 MHz, D2O)δ4.33–4.27 (m, 2H), 4.25–4.11 (m, 12H), 4.08(dd, J = 8.6, 4.9 Hz, 1H), 3.95–3.89 (m, 2H), 3.89–3.85 (m, 1H), 3.84–3.73(m, 3H), 2.89 (t, J = 7.6 Hz, 4H), 2.39–2.22 (m, 8H), 2.04–1.86 (m, 20H,AcOH), 1.86–1.63 (m, 5H), 1.63–1.46 (m, 7H), 1.43–1.32 (m, 4H), 1.31–1.27 (m,12H), 1.13 (d, J = 6.3 Hz, 3H), 0.81 (d, J = 6.2 Hz, 3H), 0.76 (d, J = 6.2Hz, 3H).

[0341] Compound 49: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Asp

[0342] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C67H114N20O27S; m / z: 555.27329 ([M+3H]3+), 832.40392 ([M+2H]2+), 1663.79211 ([M+H]+).

[0343] Compound 50: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Glu

[0344] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C68H116N20O27S; m / z: 559.94388 ([M+3H]3+), 839.40987 ([M+2H]2+), 1677.80026 ([M+H]+).

[0345] Compound 51: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Leu

[0346] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C69H119N19O26S; m / z: 554.95315 ([M+3H]3+), 831.92378 ([M+2H]2+).

[0347] Compound 52: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Gln-Ala

[0348] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C66H113N19O26S; m / z: 540.93567 ([M+3H]3+), 810.89807 ([M+2H]2+).

[0349] Compound 53: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Lys-Gln-Asp

[0350] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C73H126N22O28S; m / z: 448.73124 ([M+4H]4+), 597.97133 ([M+3H]3+), 896.45075 ([M+2H]2+), 1791.88031 ([M+H]+).

[0351] Compound 54: Ala-Glu-Pro-Val-Pro-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0352] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C67H111N17O24; m / z: 513.60834 ([M+3H]3+), 769.90656 ([M+2H]2+), 1538.80615 ([M+H]+).

[0353] Compound 55: Pro-Ala-Ala-Glu-Pro-Val-Pro-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0354] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C75H123N19O26; m / z: 569.63794 ([M+3H]3+), 853.95032 ([M+2H]2+), 1706.89331 ([M+H]+).

[0355] Compound 56: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Ala-Lys-Ala-Glu

[0356] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C64H110N18O25S; m / z: 521.93003 ([M+3H]3+), 782.38948 ([M+2H]2+), 1563.76203 ([M+H]+).

[0357] Compound 57: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Pro-Lys-Ala-Glu (acetate)

[0358] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C66H112N18O25S; m / z: 530.60034 ([M+3H]3+), 795.39502 ([M+2H]2+), 1589.78271 ([M+H]+).

[0359] 1H NMR (600 MHz, D2O) δ4.49 (dd, J = 9.1, 5.2 Hz, 1H), 4.42 (dd, J =8.4, 5.9 Hz, 1H), 4.31 (q, J = 7.4, 6.4 Hz, 2H), 4.26–4.13 (m, 9H), 4.11–4.05(m, 2H), 3.94–3.86 (m, 2H), 3.83–3.69 (m, 3H), 3.55–3.47 (m, 1H), 2.93–2.87(m, 4H), 2.54–2.42 (m, 2H), 2.36–2.16 (m, 9H), 2.04–1.93 (m, 9H), 1.93–1.49(m, 28H, AcOH), 1.43–1.32 (m, 4H), 1.31–1.25 (m, 12H), 0.82 (d, J = 5.7 Hz, 3H), 0.76 (d, J = 5.7 Hz, 3H).

[0360] Compound 58: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Lys-Gln-Glu

[0361] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C74H128N22O28S; m / z: 452.23230 ([M+4H]4+), 602.64038 ([M+3H]3+), 903.45508 ([M+2H]2+), 1805.90295 ([M+H]+).

[0362] Compound 59: Arg-Lys-Asp-Val-Tyr-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu

[0363] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C68H112N20O24; m / z: 399.21298 ([M+4H]4+), 531.94670 ([M+3H]3+), 797.41529 ([M+2H]2+), 1593.81299 ([M+H]+).

[0364] Compound 60: Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ser-Glu (acetate)

[0365] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C63H108N18O26S; m / z: 522.58909 ([M+3H]3+), 783.37789 ([M+2H]2+), 1565.73801 ([M+H]+).

[0366] 1H NMR (600 MHz, D2O) δ4.45–4.39 (m, 1H), 4.37 (t, J = 5.5 Hz, 1H), 4.34–4.26 (m, 2H), 4.25–4.06 (m, 10H), 3.91–3.90 (m, 2H), 3.88–3.86 (m, 1H), 3.85–3.80 (m, 2H), 3.79–3.74 (m, 3H), 2.89 (t, J = 7.5 Hz, 4H), 2.54–2.43 (m,1H), 2.39–2.22 (m, 7H), 2.06–1.96 (m, 8H), 1.95–1.92 (m, 23H, AcOH), 1.91–1.87 (m, 2H), 1.86–1.73 (m, 2H), 1.72–1.65 (m, 1H), 1.62–1.54 (m, 6H), 1.54–1.48 (m, 1H), 1.45–1.31 (m, 4H), 1.30 (d, J = 7.2 Hz, 9H), 0.82 (d, J = 5.9Hz, 3H), 0.77 (d, J = 5.9 Hz, 3H).

[0367] Compound 61: Glu-Ala-Glu-Ala-Ala-Gly-Lys-Ala-Glu (acetate)

[0368] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C35H58N10O16; m / z: 438.21012 ([M+2H]2+), 875.40922 ([M+H]+).

[0369] 1H NMR (600 MHz, D2O)δ4.29–4.15 (m, 7H), 4.11 (dd, J = 8.6, 4.9 Hz,1H), 3.95 (t, J = 6.5 Hz, 1H), 3.87–3.77 (m, 2H), 2.88 (t, J = 7.6 Hz, 2H),2.39–2.25 (m, 7H), 2.10–1.94 (m, 9H, AcOH), 1.91–1.78 (m, 2H), 1.78–1.69 (m,1H), 1.69–1.62 (m, 1H), 1.62–1.53 (m, 2H), 1.37–1.30 (m, 2H), 1.30–1.25 (m, 14H).

[0370] Compound 62: Phe-Asp-Ala-Leu-Lys-Gln-Gln-Phe-Gln-Ala-Phe-Gln-Leu-Glu

[0371] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 571.62579 ([M+3H]3+), 856.93494 ([M+2H]2+)

[0372] Compound 63: His-Cys-Leu-Ala-Gly-Leu-Lys-Lys-Asp-Leu-Glu-Asp-Leu-Glu

[0373] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 528.60779 ([M+3H]3+), 792.40802 ([M+2H]2+)

[0374] Compound 54: Glu-Ile-Asn-Gln-Leu-Glu-Leu-Ile-Lys-Gln-Ala-Ser-Ile

[0375] High-resolution mass spectrometry (Orbitrap Exploris) m / z 500.62137 ([M+3H]3+), 749.92682 ([M+2H]2+), 1499.85291 ([M+H]+)

[0376] Compound 65: Ala-Ala-Arg-Leu-Ala-Asp-Glu-Leu-Arg-Ala-Glu

[0377] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 405.55411 ([M+3H]3+), 607.82733 ([M+2H]2+), 1214.64624 ([M+H]+)

[0378] Compound 66: Glu-Thr-Leu-Gln-Arg-Lys-Asn-Lys-Glu

[0379] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 382.54669 ([M+3H]3+), 573.31622 ([M+2H]2+), 1146.61548 ([M+H]+)

[0380] Compound 67: Val-Asp-Ala-Ala-Val-Leu-Glu-Lys-Leu-Glu

[0381] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 543.80505 ([M+2H]2+), 1086.60242 ([M+H]+)

[0382] Compound 68: Ala-Ala-Val-Leu-Asp-Lys-Leu-Glu

[0383] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 429.74985 ([M+2H]2+), 858.49243 ([M+H]+)

[0384] Compound 69: Ala-Ala-Val-Leu-Glu-Lys-Leu-Glu

[0385] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 436.75772 ([M+2H]2+), 872.50806 ([M+H]+)

[0386] Compound 70: Lys-Phe-Tyr-Ser-Gln-Ser-Thr-Ala-Ser-Ser-Ser-Tyr-Ala-Tyr-Pro-Ser-His-Phe -Gly-Pro-Ala-Gly-Phe-Ser-Gly-Ser-His-Ser

[0387] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 736.58075 ([M+4H]4+), 981.77148 ([M+3H]3+), 1472.15320 ([M+2H]2+)

[0388] Compound 71: Val-Asp-Ala-Ala-Val-Ile-Glu-Lys-Ile-Glu

[0389] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 543.80481 ([M+2H]2+), 1086.60217 ([M+H]+)

[0390] Compound 72: Val-Asp-Ala-Ala-Met-Val-Leu-Leu-Thr-Arg

[0391] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 544.80920 ([M+2H]2+), 1088.61121 ([M+H]+)

[0392] Compound 73: Val-Asp-Ala-Ala-Val-Leu-Met-Leu-Arg-Thr

[0393] High-resolution mass spectrometry (Orbitrap Exploris) m / z: 544.80945 ([M+2H]2+), 1088.61169 ([M+H]+)

[0394] Example 2: FGFR receptor binding assay: FGFR1 protein target affinity test

[0395] The affinity of the peptide compound prepared in this invention for FGFR1 was tested using an FGFR1 allosteric inhibitor (Sigma product number SSR128129E) as a positive control.

[0396] FGFR1 affinity screening was performed as follows: an appropriate concentration of FGFR1 solution was taken and an appropriate concentration of positive compound SSR128129E (Sigma) was added. Then, the sample solution of the polypeptide compound of the present invention, which was lyophilized into powder and dissolved in protein buffer, was added. The mixture was incubated at room temperature for 50 min, and then ultrafiltration was performed to screen the solutions with different treatments. An appropriate volume of acetonitrile was added to each solution to precipitate the protein. After centrifugation, the supernatant was collected for mass spectrometry analysis.

[0397] Affinity screening conditions: One-dimensional liquid chromatography (LC) conditions: PolyLC column, mobile phase A: KH₂PO₄, NaCl, pH 7.5, mobile phase B: acetonitrile, column temperature 8℃, flow rate 1 mL / min, appropriate injection volume; Two-dimensional liquid chromatography (LC) conditions: BONUSRRHD column, mobile phase A: 0.1% formic acid aqueous solution, mobile phase B: 0.1% formic acid acetonitrile, column temperature 60℃, flow rate 0.3 mL / min, injection volume 40 μL. Gradient elution was used: 98% A (0–1 min), 98%–10% A (1–6 min), 10% A (6–7.5 min), 10%–98% A (7.5–8 min), 98% A (8–9.5 min). Mass spectrometry conditions: Agilent 6530 Q-Tof, ESI ion source, voltage 3.5 kV, mass-to-charge ratio scan range m / z = 200-3000, acquisition mode positive ion scan, ion source temperature 350℃, desolventization temperature 300℃, nitrogen flow rate 8 L / min.

[0398] Affinity screening revealed that compounds 1 and 62-73 have an affinity for FGFR1.

[0399] Table 4. Screening results of peptide compounds that bind with affinity to FGFR1

[0400]

[0401] * The Cb / Ct ratio refers to the amount of the compound in the lower layer (mass spectrometry signal intensity) compared to the amount of the compound in the upper layer during the ultracentrifugation affinity screening. The larger the Cb / Ct ratio, the stronger the affinity.

[0402] Example 3: Anti-ulcer effect of the polypeptide compound of the present invention on an ethanol-induced mouse gastric ulcer model

[0403] Experimental animals: Male C57BL / 6JGPt mice at 6 weeks old, weighing about 20 g per mouse, with 10 mice in each group. Chengdu Yakang Biotech Co., Ltd., Animal License No.: SCXK (Chuan) 2020 - 034.

[0404] Experimental method:

[0405] After grouping the experimental animals, they were administered drugs according to Table 5 one day before the experiment. Among them, the model group was given 200 μl of pure water, and the remaining drug-administered groups were given 200 μl of the drug solution. Then, all animals started fasting but not water-deprived for 24 h.

[0406] One hour after drug administration on the second day, each group of mice was intragastrically administered 9 μl / g of absolute ethanol to establish the model. One hour later, the animals were sacrificed by cervical dislocation. The gastric cardia was ligated and the pylorus was clamped, and the whole stomach was removed. 1 mL of 1% formaldehyde solution was injected into the gastric body through the glandular stomach. The cardia was ligated, and the stomach was immediately placed in 1% formaldehyde solution after removal. After soaking for 30 min, the gastric tissue was taken out, cut along the greater curvature of the stomach, and the gastric contents were rinsed clean with physiological saline. After spreading out, the damage of the mouse gastric mucosa was observed and measured, the ulcer index and ulcer inhibition rate were calculated, and a panoramic photo of the stomach was taken; finally, the gastric tissue was fixed by immersing the tissue sample in formalin.

[0407] Calculation method of ulcer index: For a strip-shaped injury with a length greater than 1 mm, measure its length and count 1 point per millimeter; if its width is greater than 1 mm, double the score according to the number of millimeters of the width; for a length less than 1 mm, count 0.5 points, and add up the scores to obtain the ulcer index of the animal.

[0408] Ulcer inhibition rate % = (ulcer index of the model group - ulcer index of the drug-administered group) / ulcer index of the model group × 100%.

[0409] Data statistical method: The ulcer index is expressed in the form of mean ± standard deviation; the ulcer inhibition rate is calculated using the median and mean values of the ulcer indices of each drug-administered group and the mock group; the data is statistically analyzed using Excel, and the difference data of the gastric ulcer index is statistically analyzed using an independent sample T-test. Table 5 shows the relative ulcer inhibition rate results of the compounds of the present invention.

[0410] Table 5. Antiulcer activity of the compounds of the present invention after a single dose in a mouse ethanol-induced model.

[0411]

[0412]

[0413] *Note:

[0414] The anti-ulcer activity of each compound was determined through multiple batches of experiments. For ease of comparison, the anti-ulcer activity is expressed as the average of the relative ulcer inhibition rate (with compound 1 as the control group in each batch of experiments), i.e., relative ulcer inhibition rate = (ulcer inhibition rate of the tested compound) / (ulcer inhibition rate of compound 1).

[0415] ND indicates that no testing has been performed.

[0416] Example 4: Pharmacodynamic study of the effect of the polypeptide compound of the present invention on the healing of acute mechanical skin injury in rats.

[0417] SPF-grade SD rats, weighing 180-230g, were housed in clean, sterilized cages. Water, feed, and bedding were provided daily at regular intervals, and the temperature was maintained at 22℃ with humidity at 55%-65%. The rats were allowed one week to acclimatize. They were then randomly assigned to three groups: a model control group (saline), a Jin Yintai control group (40 IU / cm², Shenzhen Huashengyuan Gene Engineering Development Co., Ltd.), and a test peptide treatment group (40 μg / cm²), with six rats in each group. After successful anesthesia with 3% sodium pentobarbital via intraperitoneal injection, the hair 1 cm from the wound edge was trimmed. The wound area was first disinfected with iodine, then locally disinfected with 75% alcohol. A 1.5 cm × 1.5 cm (diameter 1.5 cm) circular full-thickness skin wound was created 4 cm below the back, near the neck, with the spine as the midline, extending to the muscle layer, to establish an acute mechanical injury animal model. After modeling, the rats were housed individually with the wound exposed. During dressing changes, the wound was first cleaned with povidone-iodine, then rinsed with sterile saline and dried. Except for the model group, 40 µL of the corresponding drug solution was applied topically to the wounds of rats in other groups once daily. On days 0, 5, 10, and 14 of drug administration, images of the wounds of each group of rats were captured. The wound area was calculated using image analysis software (Image J), ​​and the wound healing rate was then determined according to the formula.

[0418] The results showed that peptides 68, 67, and 1 all tended to promote wound healing. The results are shown in Table 6.

[0419] Table 6: Effects of representative compounds of this invention on promoting wound healing

[0420]

[0421] Note: * indicates that p < 0.05 compared to the model group.

[0422] Example 5: Therapeutic effect of compound 1 on a mouse model of chronic atrophic gastritis

[0423] Methods: Chronic atrophic gastritis in mice was modeled using MNNG (N-methyl-N-nitro-N-nitrosoguanidine) combined with ranitidine. Mice were allowed free access to an aqueous solution containing MNNG (100 mg / ml), while simultaneously receiving ranitidine (8 mg / ml) via gavage at a fixed time each day for 20 weeks. After 20 weeks of modeling, in addition to drinking ordinary distilled water, mice were administered compound 1 (5 mg / kg) via gavage daily. The therapeutic effect of compound 1 on chronic atrophic gastritis was observed after 2 weeks of administration.

[0424] Results: Histological staining showed that the gastric glandular structure in the gastric body and antrum of the model group mice was disordered, accompanied by a decrease in the number of gastric acid cells (H+-K+-ATPase positive) and a decrease in the mucosal epithelial height of the gastric antrum. After 2 weeks of treatment with compound 1, compared with the model group, the gastric gland structure of mice in the compound 1 group returned to normal, the number of gastric acid cells increased significantly, and the mucosal height of the gastric antrum basically returned to normal. (See attached results). Figure 15 These results indicate that compound 1 can promote the repair of chronic atrophic gastritis in mice.

[0425] Example 6: The promoting effect of the polypeptide compound of the present invention on the proliferation of HaCAT cells

[0426] Experimental Methods: Human immortalized keratinocytes (HaCaT cells) were passaged at a concentration of 1.0×10⁵–5.0×10⁵ / mL and cultured at 37℃ and 5% CO₂ for 24–36 hours for biological activity assay. Cells were digested with 0.25% trypsin for 5 min, and digestion was terminated by adding at least one volume of 1640 whole blood medium. The cell suspension was collected, centrifuged at 1000 RPM for 3 min, the supernatant was discarded, and the cells were resuspended in 2 mL of 1640 whole blood medium. 20 μL of the cell suspension was stained with AOPI, and the cell concentration was determined using a cell counter. A 5×10⁴ / mL concentration of 1640 medium with 10% serum was prepared and seeded into 96-well cell culture plates at 100 μL per well (5000 cells / well), and cultured overnight at 37℃ and 5% CO₂.

[0427] After 24 h, the original culture medium was discarded, and 100 μL of 1% serum-concentrated 1640 medium was added to prepare solutions of different concentrations of polypeptide compounds, bringing the final concentrations of the tested polypeptide compounds to 0.2, 0.4, and 0.8 μg / mL, respectively. An EGF control group was set up, containing 100 μL of recombinant human epidermal growth factor (EGF) solution prepared with 1% serum-concentrated 1640 medium, resulting in a final concentration of 100 ng / mL. A model control group was prepared by adding an equal volume of 1% serum-concentrated 1640 medium. Cells were cultured at 37℃ and 5% CO2 for 24 h, 48 h, and 72 h, and the proliferation of the HaCaT cell line was detected using a CCK8 assay kit.

[0428] The test results are as follows Figures 16 to 27 As shown, peptide compounds 74, 73, 72, 71, 70, 69, 68, 67, 66, 65, 1, 64, and 63 exhibited proliferative effects on HaCaT cells at low concentrations (i.e., 0.2–0.8 μg / mL). (* indicates P < 0.05; ** indicates P < 0.01).

[0429] The results showed that most of the H25 peptides of the present invention had a significant proliferative effect on HaCaT cells, and the effect was significantly better than that of the EGF group (100 ng / mL).

[0430] Example 7: The promoting effect of the polypeptide compound of the present invention on the proliferation of HMEC-1 cells

[0431] Human microvascular endothelial cells (HMEC-1 cells) were cultured in 10% serum at 37°C and 5% CO2, with the medium changed every 1–2 days, and passaged at a cell concentration of 4 × 10⁶ cells / mL. Cells were collected and prepared into a culture medium at 5 × 10⁴ cells / mL; 100 μL per well (5000 cells / well) were seeded into 96-well cell culture plates and cultured at 37°C and 5% CO2 until cell adhesion occurred. The polypeptide compound of this invention was prepared into test concentrations (0.05 μg / mL, 0.2 μg / mL, 0.8 μg / mL) in 0% serum medium as the experimental group. A positive control (300 ng / mL) was prepared using the same method and added as the positive control. An equal volume of 0% serum-free, drug-free medium was added to the control group. All wells were incubated in duplicate at 37°C and 5% CO2 for 48 hours. Cell proliferation was detected using CCK-8: the old culture medium was removed, and serum-free culture medium containing 10% CCK-8 was added to each well. After incubation in an incubator for 2 h, the absorbance was measured at 450 nm using an ELISA reader.

[0432] The test results are as follows Figures 28 to 32 As shown, peptide compounds 74, 72, and 1 all exhibited significant proliferative effects on HMEC-1 cells. In the figure, * indicates promotion (P < 0.05); ** indicates promotion (P < 0.01).

[0433] Example 8: Effect of peptides on RSC96 cell proliferation

[0434] Glial cells (RSC96 cells) were adjusted to a concentration of 1.0 × 10⁵–5.0 × 10⁵ / mL and passaged for 24–36 hours at 37°C and 5% CO₂ for biological activity assay. Cells were digested with trypsin and collected, and seeded into 96-well cells at a concentration of 5 × 10⁴ / mL in serum-free medium, at a density of 100 μL per well (5000 cells / well), and cultured overnight at 37°C and 5% CO₂.

[0435] The polypeptide compounds of the present invention were prepared into test concentrations (0.2ug / ml, 0.4ug / ml, 0.8ug / ml) using 0% serum culture medium as experimental groups. An equal volume of 0% serum-free culture medium was added to the control group. The mixture was added in 4 replicates and incubated at 37°C and 5% CO2 for 48 hours.

[0436] Cell proliferation was detected using CCK-8: the old culture medium was removed, and serum-free culture medium containing 10% CCK-8 was added to each well. After incubation in an incubator for 2 hours, the absorbance was measured at 450 nm using a microplate reader.

[0437] The test results are as follows Figures 33 to 40 As shown, peptide compounds 74, 68, 66, 1, and 64 all exhibited significant proliferative effects on RSC96 cells. In the figure, * indicates P < 0.05 compared to the control group; ** indicates P < 0.01 compared to the control group.

[0438] Although the present invention has disclosed the above embodiments, the implementation of the present invention is not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, or simplifications that do not depart from the present invention should be considered as equivalent substitutions and are included within the protection scope of the present invention.

Claims

1. A compound or a physiologically compatible salt thereof, wherein the compound is selected from: Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys; Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Met(O) - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Val - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Gln; Ser - Met - Gln - Ala - Ser - Ala - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Ala - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Leu - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Ala - Gly - Lys - Ala - Glu; Ser - Met - Ala - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Ile - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Asp; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Asn; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Ala - Ala - Glu; Ser - Met - Gln - Ala - Ser - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Ala; Ser - Met - Gln - Ala - Ala - Leu - Glu - Ala - Glu - Ala - Lys - Gly - Lys - Ala - Glu; Ala-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-His-Ala-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-His-Gly-Lys-Ala-Glu; Ser-Met-Gln-Ala-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Ser-Met-Gln-Ala-Ser-Val-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Glu-Ala-Glu-Ala; Thr-Ala-Ser-Thr-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Ser-Thr-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Ala-Glu-Pro-Val-Pro-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Ala-Lys-Ala-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Pro-Lys-Ala-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ala-Lys-Gln-Glu; Ser-Met-Gln-Ala-Ser-Leu-Glu-Ala-Glu-Ala-Lys-Gly-Lys-Ser-Glu; Glu-Ala-Glu-Ala-Ala-Gly-Lys-Ala-Glu.

2. Use of the compound of claim 1 or a physiologically compatible salt thereof in the preparation of a medicament for repairing gastric ulcers.

3. A pharmaceutical composition comprising the compound of claim 1 or a physiologically compatible salt thereof and a physiologically acceptable carrier.

4. A daily chemical composition comprising the compound of claim 1 or a physiologically compatible salt thereof and a physiologically acceptable carrier.

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