Polypeptide compound and application thereof in treating enteritis

Abstract

The invention relates to a novel polypeptide and application thereof. The polypeptide provided by the invention is short in peptide chain and faster and better in absorption. Experimental results show that the polypeptide provided by the invention shows an obvious enteric cavity dilatation relieving effect in pharmacodynamic tests of a zebra fish inflammatory bowel disease model and a rat inflammatory bowel disease model. The polypeptide provided by the invention has a remarkable colitis relieving effect, and can be used for preparing a medicine for treating enteritis, especially a medicine for treating ulcerative colitis.

Description

Technical Field

[0001] This invention relates to a novel polypeptide and its applications. The polypeptide of this invention has significant therapeutic effects on inflammatory bowel disease, especially on ulcerative colitis. Background Technology

[0002] Enteritis is an inflammatory reaction of the intestines caused by various factors, such as infection by pathogens like bacteria and viruses, immune system damage, radiation damage, dietary irritants, and drug irritants. Clinical manifestations mainly include fever, nausea, vomiting, abdominal pain, diarrhea, watery stools or mucus-containing bloody stools, and some patients may also experience tenesmus.

[0003] Enteritis can be classified into specific and nonspecific enteritis based on its inflammatory manifestations. Specific enteritis includes inflammatory bowel disease (IBD), necrotizing bowel disease (NBD), and dysbiosis-related enteritis. Among these, IBD, a relatively common condition, is an idiopathic, non-infectious, chronic, persistent, relapsing inflammatory disease affecting the intestines. Clinical manifestations include abdominal pain and diarrhea, bloody or purulent stools, and even fever, abdominal masses, weight loss, and malnutrition. IBD is a chronic, long-term disease requiring long-term, aggressive treatment.

[0004] Generally, specific enteritis has a clear cause, such as bacterial enteritis, pseudomembranous enteritis, viral enteritis, and radiation enteritis. Nonspecific enteritis, on the other hand, lacks a clear cause and is considered to be caused by immune system diseases, such as ulcerative colitis and Crohn's disease. Inflammatory bowel disease includes ulcerative colitis and Crohn's disease. Ulcerative colitis is a chronic nonspecific inflammatory bowel disease with an unclear etiology, most commonly occurring in young adults. Clinical manifestations include persistent or recurrent diarrhea, bloody and mucous stools accompanied by abdominal pain, tenesmus, and varying degrees of systemic symptoms. Extraintestinal manifestations may occur, affecting the skin, mucous membranes, joints, eyes, liver, and gallbladder. Complications include toxic megacolon, intestinal perforation, lower gastrointestinal bleeding, intraepithelial neoplasia, and cancer. Accurate incidence data for ulcerative colitis (UC) in my country are lacking; regional epidemiological surveys suggest an incidence of 0.42-2.22 per 100,000. Although the incidence rate remains low compared to Western countries, it has shown a significant upward trend compared to 20 years ago. The lesions primarily affect the colonic mucosa and submucosa, starting from the distal colon and progressing proximally, sometimes involving the entire colon; 5% may involve the terminal ileum, exhibiting a continuous distribution. The main clinical manifestations of ulcerative colitis are diarrhea, abdominal pain, and bloody, mucous stools. The etiology is not fully understood, but it is currently believed to be related to infection, immune abnormalities, genetics, and psychological factors. Furthermore, patients with enteritis often experience symptoms such as intestinal dilatation (also known as intestinal megacolon or bowel dilatation), possibly due to inflammation disrupting the neural and muscular regulatory mechanisms controlling normal bowel function, causing intraluminal pressure to dilate the intestinal wall beyond its normal range of motion. Alternatively, it may be caused by microbial overgrowth and the toxins produced, as well as mucus exudation. Relieving or inhibiting intestinal dilatation is helpful in the treatment of enteritis.

[0005] While commonly used medications for ulcerative colitis have some preventative and therapeutic effects, they still suffer from drawbacks such as slow absorption, long treatment cycles, and unclear therapeutic effects. Summary of the Invention

[0006] To overcome the shortcomings and defects of existing technologies, the present invention aims to provide a novel polypeptide compound and its applications. The inventors of this invention have discovered that the polypeptide compound of this invention can effectively or significantly alleviate or inhibit intestinal distension, thereby playing a role in the treatment of enteritis.

[0007] In a first aspect, the present invention relates to compounds of formula (I) or physiologically compatible salts thereof, wherein the compounds of formula (I) are as follows:

[0008] HX a -Z1-Z2-Z3-Z4-X b -OH(I)

[0009] in

[0010] Z1 is Pro, Ala, Gly, D-Pro, or missing;

[0011] Z2 is Val, Ala, D-Val, Ile, or missing;

[0012] Z3 is Pro, Ala, Gly, D-Pro, or missing;

[0013] Z4 is Gln, Ala, Glu, Ile, D-Gln or deleted;

[0014] The condition is that at most two of Z1, Z2, Z3 and Z4 are missing.

[0015] X a A sequence containing 0-10 amino acid residues;

[0016] X b It is a sequence containing 0-9 amino acid residues.

[0017] In one implementation, two of Z1, Z2, Z3, and Z4 are missing. In one implementation, Z1 and Z2 are missing. In one implementation, Z3 and Z4 are missing. In one implementation, one of Z1, Z2, Z3, and Z4 is missing. In one implementation, Z1 is missing. In one implementation, Z4 is missing. In one implementation, none of Z1, Z2, Z3, and Z4 are missing.

[0018] In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is D-Pro; Z2 is Val; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Ala; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Ile; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Pro; and Z4 is missing. In one embodiment, Z1 is Pro; Z2 is Ala; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is Gly; Z2 is Val; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Gly; and Z4 is Gln. In one embodiment, Z1 is Ala; Z2 is Val; Z3 is Pro; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Pro; and Z4 is Glu. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is D-Pro; and Z4 is Gln. In one embodiment, Z1 is Pro; Z2 is Val; Z3 is Pro; and Z4 is D-Gln.

[0019] In one implementation scheme, X a For X a10 -X a9 -X a8 -X a7 -X a6 -X a5 -X a4 -X a3 -X a2 -X a1 -*, where * indicates the position connecting to Z1-Z2-Z3-Z4,

[0020] X a10 Gly or missing,

[0021] X a9 For Gly, Pro, Ser or missing,

[0022] X a8 For Pro, Glu, Ser or missing,

[0023] X a7 For Arg, Glu, Thr, Ser or missing,

[0024] X a6 For Lys, Thr, Ala, Asp, Glu, Arg, Ser, or missing,

[0025] X a5 For Asp, Ala, Val, Arg, Phe, Ile, Pro, Lys, Glu or missing,

[0026] X a4 Val, Phe, Pro, Pyro-Glu, Lys, Leu, Ile, Ala, Asp, or missing.

[0027] X a3 Tyr, Leu, Pro, Asp, Arg, Glu, Lys, Tys, Val, Ile, or missing.

[0028] X a2 For D-Lys, Lys, Ala, Arg, Val, Glu, Tyr or missing, and

[0029] X a1 For Glu, Ala, D-Glu, Tyr, Gln, Asp, Asn or missing.

[0030] In one implementation scheme, X a For X aa -X a2 -X a1 -*, where * indicates the position connecting to Z1-Z2-Z3-Z4, X a2 -X a1 For Lys-Glu-*, Arg-Glu-*, Val-Tyr-*, Glu-Glu-*, Lys-Gln-*, Lys-D-Glu-*, or Tyr-Glu-*, and X aa For X a10 -X a9 -X a8 -X a7 -X a6 -X a5 -X a4 -X a3 -, where X a10 For Gly or missing, X a9 For Gly, Pro, Ser, or missing, X a8 For Pro, Glu, Ser, or missing, X a7 For Arg, Glu, Thr, Ser or missing, X a6 For Lys, Thr, Ala, Asp, Glu, Arg, Ser, or missing, X a5 For Asp, Ala, Val, Arg, Phe, Ile, Pro, Lys, Glu or missing, X a4Val, Phe, Pro, Pyro-Glu, Lys, Leu, Ile, Ala, Asp or missing, and X a3 Tyr, Leu, Pro, Asp, Arg, Glu, Lys, Tys, Val, Ile, or missing. In one implementation, X aa Arg-Lys-Asp-Val-Tyr-, Gly-Pro-Glu-Thr-Ala-Phe-Leu-, Val-Pro-Pro-, Pyro-Glu-Leu-, Arg-Lys-Asp-, Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-, Ser-Ser-Glu-Asp-Ile-Lys-Glu-, Ser-Ser-Glu-Asp-Ile-Lys-, Val-Pro-Tys-, Pro-Ala-Tys-, Arg-Lys-Asp-Val-, Ser-Ser-Glu-Asp-Ile- or missing, where the rightmost connector - indicates a hyphen with -X. a2 -X a1 Connection. In one implementation, X a For X aa -X a2 -X a1 -*, where * indicates the position connecting to Z1-Z2-Z3-Z4, where X a2 -X a1 For Lys-Glu-*, and X aa As defined above.

[0031] In one implementation scheme, X a For X a1 -*, where * indicates the position connecting to Z1-Z2-Z3-Z4, where X a1 For Glu, Ala, D-Glu, Asp, Asn or missing.

[0032] In one implementation scheme, X a For X a2 -X a1 -*, where * indicates the position connecting to Z1-Z2-Z3-Z4, where X a2 For Lys, and X a1 It can be Glu, D-Glu, or Gln.

[0033] In one implementation scheme, X aFor Glu-*, Ala-*, D-Glu-*, Asp-*, Asn-*, Lys-Glu-*, D-Lys-Glu-*, Ala-Glu-*, Lys-Gln-*, Lys-D-Glu-*, Lys-Ala-*, Arg-Lys-Asp-Val-Tyr-L ys-Glu-*, Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Glu-*, Val-Pro-Pro-Lys-Glu-*, Pyro-Glu-Leu-Lys-Glu-*, Arg-Lys-Asp-Val-Tyr-*, Gly- Pro-Glu-Thr-Ala-Phe-Leu-Arg-Lys-Glu-*, Ser-Ser-Glu-Asp-Ile-Lys-Glu-Lys-Glu-*, Ser-Ser-Glu-Asp-Ile-Lys-Glu-Glu-*, Val-Pro-Tys-Lys-Glu-*, Pro-Ala-Tys-Lys-Glu-*, Arg-Lys-Asp-Val-Tyr-Glu-*, Ser-Ser-Glu-Asp-Ile-Lys-Glu-* or missing, where * indicates the position connected to Z1-Z2-Z3-Z4.

[0034] In one implementation scheme, X b For **-X b1 -X b2 -X b3 -X b4 -X b5 -X b6 -X b7 -X b8 -X b9 Where ** indicates the position connecting to Z1-Z2-Z3-Z4,

[0035] X b1 For Ala, D-Ala, or missing,

[0036] X b2 For Lys, Ala, D-Lys, Arg, or missing,

[0037] X b3 For Pro, Gly, D-Pro, Ser, Val, Ala or missing,

[0038] X b4 For Arg, Ala, Ser, Pro, D-Arg or missing,

[0039] X b5For Lys, Ala, D-Lys, Glu, Tyr or missing,

[0040] X b6 Val, Asp, D-Val, Ala, or missing.

[0041] X b7 For Ala, D-Ala, Ile, or missing,

[0042] X b8 For Ala, D-Ala, Lys, or missing, and

[0043] X b9 It is Gln, Ala, Glu, Asn, D-Gln or missing.

[0044] In one implementation scheme, X b For **-X b1 -X b2 -X b3 -X b4 -X b5 -X b6 -X b7 -X b8 -X b9 Where ** indicates the position connecting to Z1-Z2-Z3-Z4,

[0045] X b1 For Ala or D-Ala,

[0046] X b2 For Lys, Ala, or D-Lys,

[0047] X b3 For Pro, Gly, or Ala,

[0048] X b4 For Arg or D-Arg,

[0049] X b5 For Lys or missing,

[0050] X b6 Val, D-Val, Ala, or missing.

[0051] X b7 Ala or missing,

[0052] X b8 For Ala or missing, and

[0053] X b9 It is Gln, Asn, D-Gln or missing.

[0054] In one implementation scheme, Xb For **-X b1 -X b2 -X bb Where ** indicates the position connecting to Z1-Z2-Z3-Z4, X b1 -X b2 - represents **-Ala-Lys-, **-Ala-Lys-, **-Ala-D-Lys-, **-Ala-Ala-, or **-D-Ala-Lys-, X bb -X b3 -X b4 -X b5 -X b6 -X b7 -X b8 -X b9 , where X b3 For Pro, Gly, D-Pro, Ser, Val, Ala or missing, X b4 For Arg, Ala, Ser, Pro, D-Arg or missing, X b5 For Lys, Ala, D-Lys, Glu, Tyr, or absence, X b6 For Val, Asp, D-Val, Ala, or missing, X b7 For Ala, D-Ala, Ile, or missing, X b8 For Ala, D-Ala, Lys, or missing, and X b9 For Gln, Ala, Glu, Asn, D-Gln or missing; or X b3 For Pro, Gly, or Ala, X b4 For Arg or D-Arg, X b5 For Lys or missing, X b6 For Val, D-Val, Ala, or missing, X b7 For Ala or missing, X b8 For Ala or missing, and X b9 It is Gln, Asn, D-Gln, or missing. In one implementation, X bb-Pro-Arg-Lys-Val, -Pro-Arg-Lys, -Pro-Arg, -Ala-Gln, -Ala-Ala, -Pro, -Lys-Val, -Val-Pro-Tyr, -Arg- Lys, -Val-Ala, -Pro-Arg-Lys-Val-Ala, -Lys-Val-Ala-Ala-Gln, -Pro-Arg-Lys-Val-Ala-Ala-Gln, -Pro-Ar g-Lys-Val-Ala-Ala-Gln, -Pro-Ala-Lys-Val-Ala-Ala-Gln, -Pro-Arg-Ala-Val-Ala-Ala-Gln, -Pro-Arg-Ly s-Val-Ala-Ala-Ala, -Gly-Arg-Lys-Val-Ala-Ala-Gln, -D-Pro-Arg-Lys-Val-Ala-Ala-Gln, -Pro-Arg-D-Ly s-Val-Ala-Ala-Gln, -Pro-Arg-Lys-Val-D-Ala-Ala-Gln, -Pro-Arg-Lys-Val-Ala-D-Ala-Gln, -Ser-Ser-Gl u-Asp-Ile-Lys-Glu, -Pro-Arg-Lys-Val-Ala-Ala-Asn, -Pro-Arg-Lys-Val-Ala-Ala-Gln, -Pro-Arg-Lys-D- Val-Ala-Ala-Gln, -Ala-Arg-Lys-Val-Ala-Ala-Gln, -Pro-Arg-Lys-Ala-Ala-Ala-Gln, -Pro-Arg-Lys-Val-Ala-Ala-Gln, -Pro-D-Arg-Lys-Val-Ala-Ala-Gln, -Pro-Arg-Lys-Val-Ala-Ala-D-Gln or missing, where the leftmost connector - indicates a symmetric relationship with X. b1 -X b2 Connection. In one implementation, X b For **-X b1 -X b2 -X bb Where ** indicates the position connecting to Z1-Z2-Z3-Z4, X b1 -X b2 -for**-Ala-Lys-,X bb As defined above.

[0055] In one implementation scheme, X b It is **-Ala-.

[0056] In one implementation scheme, X b**-Ala-, **-Ala-Lys, **-Ala-D-Lys-, **-Ala-Ala-, **-D-Ala-Lys, **-Ala-Arg, **-Ala-Lys-Pro-Arg-Lys-Val, **-Ala-Lys-Pro-Arg-Lys, **-Ala-Lys-Pro-Arg, **-Val-Ala-Ala-Gln, **-Lys-Val-Ala-Ala, **-Ala-Lys-Pro, **-Pro-Arg-Lys-Val, **-Ala-Lys-Val-Pro-Tyr, **-Lys-P ro-Arg-Lys、**-Arg-Lys-Val-Ala、**-Ala-Lys-Pro-Arg-Lys-Val-Ala、**-Pro-Arg-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-Pro-Arg-Lys-Val-Al a-Ala-Gln、**-Ala-Ala-Pro-Arg-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-Pro-Ala-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-Pro-Arg-Ala-Val-Ala-Ala -Gln、**-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Ala、**-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-D-Pro-Arg-Lys-Val-Ala-Ala-G ln、**-Ala-Lys-Pro-Arg-D-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-Pro-Arg-Lys-Val-D-Ala-Ala-Gln、**-Ala-Lys-Pro-Arg-Lys-Val-Ala-D-Ala -Gln、**-Ala-Lys-Ser-Ser-Glu-Asp-Ile-Lys-Glu、**-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Asn、**-Ala-D-Lys-Pro-Arg-Lys-Val-Ala-Ala-G ln、**-Ala-Lys-Pro-Arg-Lys-D-Val-Ala-Ala-Gln、**-Ala-Lys-Ala-Arg-Lys-Val-Ala-Ala-Gln、**-Ala-Lys-Pro-Arg-Lys-Ala-Ala-Ala-Gln、**-D-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln,**-Ala-Lys-Pro-D-Arg-Lys-Val-Ala-Ala-Gln,**-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-D-Gln or missing, where ** indicates the position connected to Z1-Z2-Z3-Z4.

[0057] In one implementation, Z1 is Pro; Z2 is Val; Z3 is Pro; and Z4 is Gln; X a For X aa -X a2 -X a1 -

[0058] *, where * represents the position connecting to Z1-Z2-Z3-Z4, where X a2 -X a1 For Lys-Glu-*, and X b For **-X b1 -X b2 -X bb Where ** indicates the position connecting to Z1-Z2-Z3-Z4, X b1 -X b2 - is for **-Ala-Lys-, where X aa and X bb As defined above.

[0059] In one embodiment, the compound is selected from:

[0060] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 1);

[0061] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys (compound 2);

[0062] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (compound 3);

[0063] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 4);

[0064] Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 5);

[0065] Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 6);

[0066] Lys-Glu-Pro-Val (compound 7);

[0067] Val-Ala-Ala-Gln (compound 8);

[0068] Glu-Pro-Val-Pro (Compound 9);

[0069] Lys-Pro-Arg-Lys (compound 10);

[0070] Ala-Lys-Pro-Arg (compound 11);

[0071] Pro-Val-Pro-Gln (compound 12);

[0072] Val-Pro-Gln-Ala (compound 13);

[0073] Pro-Gln-Ala-Lys (compound 14);

[0074] Arg-Lys-Val-Ala (compound 15);

[0075] Lys-Val-Ala-Ala (compound 16);

[0076] Gln-Ala-Lys-Pro (compound 17);

[0077] Pro-Arg-Lys-Val (compound 18);

[0078] Ala-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 19);

[0079] Lys-Ala-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 20); Lys-Glu-Ala-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 21); Lys-Glu-Pro-Val-Ala-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 22); Lys-Glu-Pro-Val-Pro-Ala-Ala-Lys-P ro-Arg-Lys-Val-Ala-Ala-Gln (compound 23); Lys-Glu-Pro-Val-Pro-Gln-Ala-Ala-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 24); Lys-Glu-Pro-Ala-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 25); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Ala-Arg-Lys-Val-Ala-Ala-Gln (compound 26); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Ala-Lys-Val-Ala-Ala-Gln (compound 27); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Ala-Val-Ala-Ala-Gln (compound 28); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Ala-Ala-Ala-Gln (compound 29); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-P ro-Arg-Lys-Val-Ala-Ala-Ala (compound 30); Lys-Glu-Gly-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 31); Lys-Glu-Pro-Val-Gly-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 32); Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (compound 33);Lys-Glu-Pro-Val-Gly-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (Compound 34); Lys-Glu-Gly-Val-Gly-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (Compound 35); Lys-Glu-Gly-Val-Pro-Gln -Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (Compound 36); Lys-Glu-Gly-Val-Gly-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (Compound 37); Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys (Compound 38);

[0080] Pro-Val-Pro-Gln-Ala (compound 39);

[0081] Pro-Val-Pro-Glu-Ala (compound 40);

[0082] Pro-Val-Pro-Ile-Ala (compound 41);

[0083] Pro-Val-Pro-Ala (compound 42);

[0084] Glu-Pro-Val-Pro-Gln-Ala (compound 43);

[0085] Pro-Val-Pro-Gln-Ala-Lys (compound 44);

[0086] Glu-Pro-Val-Pro-Gln-Ala-Arg (compound 45);

[0087] Ala-Pro-Val-Pro-Gln-Ala-Lys (compound 46);

[0088] Glu-Pro-Val-Pro-Ala-Ala-Lys (compound 47);

[0089] Glu-Pro-Val-Pro-Gln-Ala-Ala (compound 48);

[0090] Asp-Pro-Val-Pro-Gln-Ala-Lys (compound 49);

[0091] Asn-Pro-Val-Pro-Gln-Ala-Lys (Compound 50);

[0092] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala (Compound 51);

[0093] Glu-Pro-Ala-Pro-Gln-Ala-Lys (compound 52);

[0094] Glu-Ala-Val-Pro-Gln-Ala-Lys (compound 53);

[0095] Glu-Pro-Val-Ala-Gln-Ala-Lys (compound 54);

[0096] Glu-Pro-Val-Pro-Glu-Ala-Lys (compound 55);

[0097] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val (Compound 56);

[0098] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (compound 57);

[0099] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro (Compound 58);

[0100] Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (compound 59);

[0101] Pro-Val-Pro-Gln-Ala-Lys-Pro (Compound 60);

[0102] D-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 61); Lys-D-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 62); Lys-Glu-D-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 63); Lys-Glu-Pro-D-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 64); Lys-Glu-Pro-Val-D-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 65);

[0103] Lys-Glu-Pro-Val-Pro-D-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 66);

[0104] Lys-Glu-Pro-Val-Pro-Gln-D-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 67);

[0105] Lys-Glu-Pro-Val-Pro-Gln-Ala-D-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 68);

[0106] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-D-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 69);

[0107] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-D-Arg-Lys-Val-Ala-Ala-Gln (compound 70);

[0108] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-D-Lys-Val-Ala-Ala-Gln (compound 71);

[0109] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-D-Val-Ala-Ala-Gln (compound 72);

[0110] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-D-Ala-Ala-Gln (compound 73);

[0111] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-D-Ala-Gln (compound 74);

[0112] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-D-Gln (compound 75);

[0113] Glu-Gly-Val-Pro-Gln-Ala-Lys (compound 76);

[0114] Glu-Pro-Val-Gly-Gln-Ala-Lys (compound 77);

[0115] D-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 78);

[0116] Glu-D-Pro-Val-Pro-Gln-Ala-Lys (compound 79);

[0117] Glu-Pro-D-Val-Pro-Gln-Ala-Lys (Compound 80);

[0118] Glu-Pro-Val-D-Pro-Gln-Ala-Lys (compound 81);

[0119] Glu-Pro-Val-Pro-D-Gln-Ala-Lys (compound 82);

[0120] Glu-Pro-Val-Pro-Gln-D-Ala-Lys (compound 83);

[0121] Glu-Pro-Val-Pro-Gln-Ala-D-Lys (compound 84);

[0122] Lys-Glu-Pro-Val-Pro (compound 85);

[0123] Glu-Pro-Val-Pro-Gln (compound 86);

[0124] Lys-Glu-Pro-Val-Pro-Gln (compound 87);

[0125] Val-Pro-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 88);

[0126] Pyro-Glu-Leu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 89);

[0127] Pro-Ala-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 90);

[0128] Arg-Lys-Asp-Val-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 91);

[0129] Arg-Lys-Asp-Val-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 92);

[0130] Arg-Lys-Asp-Val-Tyr-Glu-Pro-Val-Pro-Gln-Ala-Lys (Compound 93);

[0131] Arg-Lys-Asp-Val-Tyr-Pro-Val-Pro-Gln (compound 94);

[0132] Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 95);

[0133] Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Glu-Pro-Val-Pro-Gln-Ala-Lys (Compound 96);

[0134] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 97);

[0135] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 98);

[0136] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Pro-Val-Pro-Gln (compound 99);

[0137] Val-Pro-Pro-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 100);

[0138] Glu-Pro-Val-Pro-Gln-Ala-Lys-Ser-Ser-Glu-Asp-Ile-Lys-Glu (compound 101);

[0139] Glu-Pro-Val-Pro-Gln-Ala-Lys-Val-Pro-Tyr (compound 102);

[0140] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Glu (compound 103);

[0141] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Asn (compound 104);

[0142] Lys-Gln-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 105);

[0143] Lys-Glu-Pro-Ile-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 106);

[0144] Val-Pro-Gln-Ala (compound 107); or

[0145] Val-Pro-Gln-Ala-Lys (compound 108).

[0146] In one embodiment, the compound is selected from:

[0147] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 1);

[0148] Lys-Glu-D-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 63);

[0149] Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Glu-Pro-Val-Pro-Gln-Ala-Lys (Compound 96);

[0150] Val-Pro-Pro-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 100);

[0151] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Asn (compound 104);

[0152] Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 6);

[0153] Pro-Val-Pro-Gln (compound 12);

[0154] Glu-Pro-Val-Ala-Gln-Ala-Lys (compound 54);

[0155] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (compound 57);

[0156] Lys-Glu-Pro-Val-Pro-Gln-Ala-D-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 68);

[0157] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-D-Val-Ala-Ala-Gln (compound 72);

[0158] Pyro-Glu-Leu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 89);

[0159] Arg-Lys-Asp-Val-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 92);

[0160] Arg-Lys-Asp-Val-Tyr-Pro-Val-Pro-Gln (compound 94);

[0161] Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 95);

[0162] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 97);

[0163] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 98);

[0164] Lys-Gln-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 105);

[0165] Lys-Glu-Pro-Ile-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 106);

[0166] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (compound 4);

[0167] Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 5);

[0168] Glu-Pro-Val-Pro (Compound 9);

[0169] Lys-Pro-Arg-Lys (compound 10);

[0170] Arg-Lys-Val-Ala (compound 15);

[0171] Lys-Glu-Pro-Ala-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 25);

[0172] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Ala-Arg-Lys-Val-Ala-Ala-Gln (compound 26);

[0173] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Ala-Ala-Ala-Gln (compound 29);

[0174] Lys-Glu-Gly-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 31);

[0175] Lys-Glu-Pro-Val-Gly-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (compound 34);

[0176] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys (compound 38);

[0177] Glu-Pro-Val-Pro-Gln-Ala-Ala (compound 48);

[0178] Asp-Pro-Val-Pro-Gln-Ala-Lys (compound 49);

[0179] Asn-Pro-Val-Pro-Gln-Ala-Lys (Compound 50);

[0180] Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala (Compound 51);

[0181] Glu-Ala-Val-Pro-Gln-Ala-Lys (compound 53);

[0182] Glu-Pro-Val-Pro-Glu-Ala-Lys (compound 55);

[0183] Lys-D-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 62);

[0184] Lys-Glu-Pro-Val-D-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 65);

[0185] Lys-Glu-Pro-Val-Pro-D-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 66);

[0186] Lys-Glu-Pro-Val-Pro-Gln-D-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 67);

[0187] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-D-Arg-Lys-Val-Ala-Ala-Gln (compound 70);

[0188] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-D-Gln (compound 75);

[0189] Glu-Pro-Val-Pro-Gln-D-Ala-Lys (compound 83);

[0190] Lys-Glu-Pro-Val-Pro (compound 85);

[0191] Glu-Pro-Val-Pro-Gln (compound 86);

[0192] Val-Pro-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 88);

[0193] Pro-Ala-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 90);

[0194] Arg-Lys-Asp-Val-Tyr-Glu-Pro-Val-Pro-Gln-Ala-Lys (Compound 93);

[0195] Ser-Ser-Glu-Asp-Ile-Lys-Glu-Pro-Val-Pro-Gln (compound 99); or

[0196] Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Glu (Compound 103).

[0197] In a second aspect, the present invention provides the use of the polypeptide compound or a physiologically compatible salt thereof in the preparation of a medicament for treating enteritis; or a method for treating enteritis, the method comprising administering a therapeutically effective amount of the polypeptide compound or a physiologically compatible salt thereof to a subject; or the polypeptide compound or a physiologically compatible salt thereof for treating enteritis.

[0198] Furthermore, enteritis includes specific enteritis and nonspecific enteritis. Specific enteritis further includes inflammatory bowel disease, necrotizing enteropathic disease, and dysbiosis-related enteritis.

[0199] Furthermore, inflammatory bowel disease includes ulcerative colitis.

[0200] Furthermore, the present invention provides pharmaceutical compositions comprising the polypeptide compound of the present invention or a physiologically compatible salt thereof, and pharmaceutically acceptable excipients.

[0201] Furthermore, drug dosage forms include tablets, capsules, solutions, powders, and pills.

[0202] The novel polypeptide provided by this invention and the beneficial effects of its applications:

[0203] The polypeptides of this invention have short peptide chains, resulting in faster and better absorption. Experimental results show that the polypeptides provided in this application significantly alleviated intestinal distension in a rat model of acute inflammatory bowel disease; they also demonstrated significant therapeutic effects in the same model. The polypeptides of this invention have significant efficacy in relieving colitis and can be used to prepare drugs for treating enteritis, especially ulcerative colitis. Attached Figure Description

[0204] Figure 1 A schematic diagram of the solid-phase synthesis steps of peptides is shown;

[0205] Figure 2 The mass spectrum of the polypeptide is shown. Detailed Implementation

[0206] 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.

[0207] As used in this application (including the appended claims), unless the context clearly indicates otherwise, singular terms such as “a” and “the” include their corresponding plural pronouns.

[0208] Unless the context clearly indicates otherwise, the term “or” is used to mean the term “and / or” and is used interchangeably with the term “and / or”.

[0209] The term "effective amount" or "therapeutic effective amount" refers to an amount of an active ingredient (such as a compound) that, when administered to a subject to treat at least one clinical symptom of a disease or disorder, is sufficient to affect such treatment of said disease, disorder, or symptom. "Therapeutic effective amount" can vary depending on the compound, the disease, disorder, and / or the symptom of the disease or disorder, the severity of the disease, disorder, and / or the age and / or weight of the subject to be treated. In any given case, the appropriate amount may be apparent to those skilled in the art or may be determined by routine experimentation. In some embodiments, "therapeutic effective amount" is the amount of at least one compound disclosed in this application and / or at least one stereoisomer thereof, and / or at least one pharmaceutically acceptable salt thereof, that is effective in "treating" (as defined above) a subject's disease or disorder. In the case of combination therapies, "therapeutic effective amount" refers to the total amount of the combination of substances used to effectively treat a disease, disorder, or symptom.

[0210] 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.

[0211] The term "intestinal dilatation" refers to the dilation of the intestines or the intestinal tract. In patients with enteritis, intestinal dilatation is caused by various factors such as intestinal damage, mucus exudation, gas or fluid accumulation in the intestinal tract, and microbial overgrowth. The intestinal tract is thicker than normal. Recurrent enteritis is commonly seen in patients with inflammatory bowel disease, which leads to intestinal dilatation, or tumors that cause intestinal dilatation and intestinal obstruction.

[0212] The abbreviations for the amino acids used in this application are as follows:

[0213] Chinese name English name abbreviation alanine Alanine Ala Arginine Arginine Arg Asparagine Asparagine Asn Aspartic acid Aspartic acid Asp Cysteine Cysteine Cys glutamine Glutamine Gln glutamic acid Glutamic acid Glu glycine Glycine Gly Histidine Histidine His Isoleucine Isoleucine Ile Leucine Leucine Leu Lysine Lysine Lys Methionine Methionine Met Phenylalanine Phenylalanine Phe proline Proline Pro Serine Serine Ser threonine Threonine Thr Tryptophan Tryptophan Trp Tyrosine Tyrosine Tyr Valine Valine Val

[0214] Table 1. English names and Chinese abbreviations of solvents, reagents, etc.

[0215]

[0216]

[0217] Example 1: Chemical Synthesis of Peptides

[0218] The peptide compounds were synthesized using a fully automated peptide synthesizer and conventional solid-phase synthesis method. The process involved resin swelling, deprotection, washing, amino acid activation, and condensation to chemically synthesize the peptides.

[0219] Taking Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (compound 1) as an example, a schematic diagram of the polypeptide solid-phase synthesis steps can be found in [link to schematic diagram]. Figure 1 .

[0220] Step 1: Preparation of fully protected peptide resin for polypeptides

[0221] (1) Resin swelling: Weigh 7.71g of 2-Chlorotrityl Chloride Resin (SD = 0.73mmol / g), add it to a synthesis tube with a sieve plate, and swell it with 100ml of dichloromethane (DCM).

[0222] (2) Preparation of Fmoc-Gln(Trt)-resin: Weigh out Fmoc-Gln(Trt)-OH and DIPEA respectively in a molar ratio of 1:1.78:4.31 and add them to the synthesis tube. Bubble and shake with nitrogen (N2) at room temperature for 1-3 hours, then dry under vacuum; then wash with dimethylformamide (DMF) 5 times, 100 ml each time, and dry the resin under vacuum.

[0223] (3) Removal of Fmoc protecting group: Add 100 ml of 20% piperidine-DMF (v / v) solution to the reactor, bubble the reaction under nitrogen for 20 min, and then dry it. Then wash it 5 times with DMF, 100 ml each time, for 3 minutes each time, and then dry it. The Fmoc removal result is detected by ninhydrin method.

[0224] (4) Amino acid pre-activation: Add 15 mmol of Fmoc-protected amino acids, 15 mmol of HOBt and 15 mmol of DIC to a 250 ml beaker, dissolve in 100 ml of DMF, stir at room temperature and set aside.

[0225] (5) Amino acid linkage: Pour the activated protected amino acid solution into the reactor and add an appropriate amount of DCM cleaning equipment. React under nitrogen bubbling at room temperature for 1–3 hours. Check the completeness of amino acid linkage using the ninhydrin method; if complete, dry the solution. Wash the resin 5 times with DMF (100 ml / time, 3 min / time), then dry. The dosage of each amino acid and condensing agent is shown in Table 2.

[0226] (6) After the first amino acid condensation is completed, repeat steps (4) and (5) to extend the peptide chain in the order of amino acids until the last amino acid is coupled.

[0227] (7) The resin peptides were washed 4 times with DMF, 150ml each time, for 3min each time; then washed 5 times with DCM, 150ml each time, for 3min each time, and then dried.

[0228] Table 2 Dosage of Amino Acids and Condensing Agents

[0229]

[0230]

[0231] Step 2: Cutting

[0232] (1) Add 100 ml of cutting agent (TFA:TIPS:H2O=95:2.5:2.5, v / v) to the synthesis tube and bubble with nitrogen for 1.5 to 3 hours.

[0233] (2) After the cleavage reaction was completed, the cleavage agent was filtered into a 250 ml round-bottom flask. After vacuum concentration to one-quarter of the original cleavage agent volume, 10 times the existing volume of methyl tert-butyl ether was added, and a white solid was precipitated. 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 sintered glass funnel and dried under nitrogen in a fume hood until the solvent evaporated and the crude peptide became a powder. 9.04 g of crude peptide was obtained, with a crude yield of 74.7%.

[0234] Step 3: Purification, salt replacement, and freeze-drying

[0235] Peptide HPLC with direct salt replacement (acetate)

[0236] A. Chromatographic parameters

[0237] Column: Dynamic axial compression column, 80*250mm; Packing material: Daisogel C18 (SP-100-8-ODS-P)

[0238] Eluent A1: 0.1M acetic acid

[0239] Eluent A2: 0.025M acetic acid – 0.1M ammonium acetate

[0240] Elution Buffer B: Acetonitrile

[0241] Flow rate: 180 ml / min

[0242] UV detection wavelength: 220nm

[0243] B. Operating Procedures

[0244] a) Dissolve the crude peptides in water and / or acetonitrile, and filter through a 0.45 μm filter membrane.

[0245] b) 95% A1 + 5% B equilibrium chromatographic column

[0246] c) Sample injection

[0247] d) 95% A2+5% B equilibrium chromatographic column

[0248] e) Gradient elution of A1 and B

[0249] f) Collect the eluent of the target peptide.

[0250] g) Rotary evaporation concentration

[0251] h) Freeze-drying

[0252] 8.26g of crude peptide was purified to obtain 4.92g of pure peptide, with a purification yield of 59.5%.

[0253] 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.

[0254] Table 3 Synthesized polypeptide compounds

[0255]

[0256]

[0257]

[0258]

[0259]

[0260]

[0261] 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.

[0262] Compound 1: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0263] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 73 H 127 N 23 O 20 ; m / z: m / z: 549.65904([M+3H] 3+ ), 823.98493([M+2H] 2+ ), 1646.96346([M+H] + ).

[0264] 1 H NMR(600MHz,D2O+CD3CN)δ4.48(dd,J=9.5,4.1Hz,1H),4.42(dd,J=9.2,4.5Hz,1H),4.32–4.27(m,1H),4 .27–4.21(m,3H),4.20–4.07(m,6H),4.00(dd,J=8.3,4.8Hz,1H),3.95(d,J=7.5Hz,1H),3.86(t,J=6.7Hz ,1H),3.77–3.62(m,3H),3.59–3.43(m,3H),3.04(t,J=6.7Hz,2H),2.87–2.78(m,6H),2.28–2.08(m,9H), 2.00–1.80(m,31H,AcOH),1.80–1.41(m,23H),1.40–1.24(m,6H),1.24–1.18(m,9H),0.87–0.73(m,12H).

[0265] Compound 2: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys (acetate)

[0266] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 57 H 100 N 18 O 15 m / z: 426.59680([M+3H]) 3+ ), 639.39021([M+2H] 2+ ), 1277.77023([M+H] + ).

[0267] 1H NMR(600MHz,DMSO-d6)δ4.57(dd,J=8.7,4.9Hz,1H),4.41–4.11(m,10H),3.83 (s,1H),3.78(t,J=6.5Hz,1H),3.54(d,J=73.5Hz,7H),3.08(d,J=7.0Hz,2H),2 .74–2.68(m,7H),2.32(d,J=9.1Hz,2H),2.23–2.17(m,2H),2.05–1.46(m,AcOH ,56H),1.43–1.22(m,7H),1.17(d,J=7.0Hz,3H),0.82(dd,J=19.6,6.7Hz,6H).

[0268] Compound 3: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (acetate)

[0269] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 51 H 88 N 16 O 14 m / z: 383.89927([M+3H]) 3+ ), 575.34276([M+2H] 2+ ), 1149.67551([M+H] + ).

[0270] 1 H NMR (600MHz, DMSO-d6) δ4.58–4.55(m,1H),4.44–4.37(m,2H),4.33(dd,J=8.4,4.2Hz,1H ),4.30–4.22(m,3H),4.16–4.12(m,2H),3.79(d,J=6.1Hz,1H),3.64–3.48(m,6H),3.08(t ,J=7.0Hz,2H),2.71(d,J=8.2Hz,5H),2.36–2.30(m,2H),2.15(t,J=7.6Hz,2H),2.09–1. 41(m,AcOH,46H),1.37–1.29(m,4H),1.16(d,J=7.1Hz,3H),0.85(dd,J=20.7,6.7Hz,6H).

[0271] Compound 4: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0272] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C40 H 69 N 11 O 12 m / z: 448.76653([M+2H]) 2+ ), 896.52058([M+H] + ).

[0273] 1 H NMR (600MHz, DMSO-d6) δ4.56 (dd, J=9.0, 4.9Hz, 1H), 4.39 (dd, J=8.3, 4.4Hz, 1H), 4. 31–4.24(m,3H),4.13(dd,J=9.1,5.1Hz,2H),3.78(dd,J=7.9,4.3Hz,1H),3.67–3.5 1(m,4H),2.75–2.70(m,4H),2.38–2.29(m,2H),2.14(t,J=7.8Hz,2H),2.04–1.47(m ,AcOH,33H),1.35–1.28(m,4H),1.19(d,J=7.2Hz,3H),0.85(dd,J=22.1,6.8Hz,6H).

[0274] Compound 5: Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0275] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 33 H 60 N 12 O9; m / z: 385.24021([M+2H]) 2+ ), 769.46971([M+H] + ).

[0276] 1 H NMR(600MHz,D2O)δ4.23–4.17(m,1H),4.15–4.07(m,4H),3.95(dd,J=8.5,4.8Hz ,1H),3.89(d,J=7.7Hz,1H),3.25–3.15(m,2H),3.00(t,J=7.0Hz,2H),2.78(t,J =7.7Hz,2H),2.30–2.22(m,1H),2.12–2.05(m,2H),1.93–1.80(m,5H),1.75(s,A cOH,10H),1.62–1.40(m,8H),1.19(d,J=7.2Hz,8H),0.73(dd,J=6.7,4.0Hz,6H).

[0277] Compound 6: Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0278] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 34 H 57 N9O 11 m / z: 768.42641([M+H]) + ).

[0279] 1 H NMR(600MHz,D2O)δ4.46(dd,J=8.4,6.4Hz,1H),4.35–4.27(m,3H),4.26–4.18(m,2H),4.09 (dd,J=8.2,5.1Hz,1H),3.84–3.77(m,1H),3.68–3.52(m,3H),2.91(t,J=7.5Hz,2H),2.39( t,J=7.2Hz,2H),2.33(t,J=7.6Hz,2H),2.27–2.17(m,2H),2.12–1.87(m,AcOH,13H),1.83– 1.72(m,3H),1.66–1.57(m,3H),1.32(dd,J=11.8,7.5Hz,5H),0.92(dd,J=12.6,6.7Hz,6H).

[0280] Compound 7: Lys-Glu-Pro-Val (acetate)

[0281] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 21 H 37 N5O7; m / z: 472.27768 ([M+H]) + ).

[0282] 1 H NMR(600MHz,D2O)δ4.43(dd,J=10.2,4.0Hz,1H),4.27(dd,J=8.3,5.3Hz,1H),3.85–3.80(m,2H),3.68–3.60(m,1H),3.59–3.46(m,1H),2.80(t,J =7.6Hz,2H),2.22–2.14(m,2H),2.10–2.06(m,1H),1.96–1.64(m,AcOH,1 1H),1.52–1.47(m,2H),1.29–1.22(m,2H),0.71(dd,J=15.3,6.9Hz,6H).

[0283] Compound 8: Val-Ala-Ala-Gln (acetate)

[0284] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 16 H 29 N5O6; m / z: 388.2187 ([M+H]) + ).

[0285] 1 H NMR(600MHz,D2O)δ4.25(q,J=7.2Hz,1H),4.17(q,J=7.2Hz,1H),4.05(dd,J=8.5,4.8Hz,1H),3.67(d,J=6.0Hz,1H),2.23–2.1 4(m,2H),2.13–2.05(m,1H),2.04–1.95(m,1H),1.85–1.76(m,1H),1.28(dd,J=7.2,5.0Hz,6H),0.90(dd,J=10.3,6.9Hz,6H).

[0286] Compound 9: Glu-Pro-Val-Pro (acetate)

[0287] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 20 H 32 N4O7; 441.23562([M+H]) + ).

[0288] 1 H NMR (600MHz, DMSO-d6) δ4.42(dd,J=8.4,4.6Hz,1H),4.28(t,J=8.3Hz,1H),4.17(dd,J=8.5,4.7Hz,1H),3.78(dd, J=8.3,4.2Hz,1H),3.64–3.45(m,4H),2.37–2.27(m,2H),2.08–1.58(m,AcOH,16H),0.87(dd,J=25.7,6.7Hz,6H).

[0289] Compound 10: Lys-Pro-Arg-Lys (acetate)

[0290] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 23 H 45 N9O5; 264.68527 ([M+2H]) 2+ ), 528.36210([M+H] + ).

[0291] 1 H NMR (600MHz, DMSO-d6) δ4.42 (dd, J=8.2, 5.2Hz, 1H), 4.23 (t, J=6.7Hz, 1H), 4. 14(dd,J=8.9,5.2Hz,1H),4.09(d,J=7.9Hz,1H),3.64(dd,J=9.9,6.6Hz,1H),3 .45–3.40(m,1H),3.07(s,2H),2.72(t,J=6.3Hz,4H),2.11–2.04(m,1H),1.92 –1.66(m,AcOH,18H),1.56–1.48(m,8H),1.45–1.38(m,2H),1.33–1.29(m,2H).

[0292] Compound 11: Ala-Lys-Pro-Arg (acetate)

[0293] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 20 H 38 N8O5; 236.15638 ([M+2H]) 2+ ), 471.30462([M+H] + ).

[0294] 1 H NMR(600MHz,D2O))δ4.55(dd,J=8.3,5.6Hz,1H),4.34(dd,J=8.4,5.9Hz,1 H),4.09–3.96(m,2H),3.82–3.74(m,1H),3.64–3.55(m,1H),3.12(t,J=6.9 Hz,2H),2.93(t,J=7.6Hz,2H),2.28–2.20(m,1H),2.04–1.83(m,3H),1.82 (s,AcOH,6H),1.79–1.60(m,6H),1.58–1.51(m,2H),1.42(d,J=7.1Hz,5H).

[0295] Compound 12: Pro-Val-Pro-Gln (acetate)

[0296] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 20 H 33 N5O6; m / z: 440.25122 ([M+H]) + ).

[0297] 1H NMR(600MHz,D2O)δ4.33–4.18(m,3H),3.97(dd,J=8.9,4.8Hz,1H),3.74–3.65(m,1H),3.53–3.46(m,1H),3.25–3.16( m,2H),2.29–2.22(m,1H),2.17–2.10(m,3H),1.99–1.66(m,AcOH,12H),0.82(d,J=6.8Hz,3H),0.76(d,J=6.7Hz,3H).

[0298] Compound 13: Val-Pro-Gln-Ala (acetate)

[0299] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 18 H 31 N5O6; m / z: 414.23575 ([M+H]) + ).

[0300] 1 H NMR(600MHz,D2O)δ4.37(t,J=7.5Hz,1H),4.15(dd,J=8.6,6.0Hz,1H),4.07(d,J=5.5Hz,1H),4.03(t,J=7.2Hz,1H),3.68–3.61(m,1H),3.55–3 .48(m,1H),2.35–2.26(m,2H),2.26–2.15(m,2H),2.04–1.74(m,AcOH,7H),1.22(d,J=7.2Hz,3H),0.98(d,J=7.0Hz,3H),0.87(d,J=6.8Hz,3H).

[0301] Compound 14: Pro-Gln-Ala-Lys (acetate)

[0302] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 19 H 34 N6O6; m / z: 222.13480 ([M+2H]) 2+ ), 443.26183([M+H] + ).

[0303] 1H NMR(600MHz,D2O)δ4.28(dd,J=8.7,6.1Hz,1H),4.25–4.15(m,2H),4.01(dd,J=8.1,5.2Hz,1H),3.34–3.22(m,2H),2.86(t,J=7. 5Hz,2H),2.37–2.29(m,1H),2.26(t,J=7.6Hz,2H),2.00–1.84(m,5H),1.73–1.64(m,1H),1.62–1.50(m,3H),1.30–1.24(m,5H).

[0304] Compound 15: Arg-Lys-Val-Ala (acetate)

[0305] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 20 H 40 N8O5; m / z: 237.16465 ([M+2H]) 2+ ), 473.32087([M+H] + ).

[0306] 1 H NMR(600MHz,D2O)δ4.28(t,J=7.3Hz,1H),4.00–3.94(m,2H),3.90(t,J=6.4Hz,1H),3.07(t,J=7.0Hz,2H),2.85(t,J=7.6Hz,2H),2.01–1. 92(m,1H),1.80–1.75(m,AcOH,8H),1.73–1.59(m,2H),1.59–1.40(m,4H),1.35–1.22(m,2H),1.19(d,J=7.2Hz,3H),0.81(t,J=6.9Hz,6H).

[0307] Compound 16: Lys-Val-Ala-Ala (acetate)

[0308] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 17 H 33 N5O5; m / z: 388.25660 ([M+H]) + ).

[0309] 1H NMR(600MHz,D2O)δ4.18(q,J=7.1Hz,1H),4.03–3.91(m,3H),2.87(t,J=7.7Hz,2H),1.98–1.89(m,1H),1.83–1.74(m, AcOH,6H),1.60–1.53(m,2H),1.36–1.27(m,2H),1.26(d,J=7.2Hz,3H),1.19(d,J=7.2Hz,3H),0.83(d,J=6.8Hz,6H).

[0310] Compound 17: Gln-Ala-Lys-Pro (acetate)

[0311] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 19 H 34 N6O6; m / z: 443.26161 ([M+H]) + ).

[0312] 1 H NMR(600MHz,D2O)δ4.48(dd,J=8.2,5.6Hz,1H),4.26–4.20(m,1H),4.10(dd,J=8.5,5.4Hz,1H),3.91(t,J=6.6Hz,1H),3.68–3.61(m,1H),3.55–3.4 3(m,1H),2.88(t,J=7.5Hz,2H),2.39–2.27(m,2H),2.15–1.96(m,3H),1. 92–1.69(m,8H),1.66–1.49(m,3H),1.43–1.29(m,2H),1.28–1.22(m,3H).

[0313] Compound 18: Pro-Arg-Lys-Val (acetate)

[0314] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 22 H 42 N8O5; m / z: 250.17158 ([M+2H]) 2+ ), 499.33464([M+H] + ).

[0315] 1H NMR(600MHz,D2O)δ4.27(dd,J=8.7,6.0Hz,1H),4.24–4.17(m,2H),3.88(d,J=6.2Hz,1H),3.33–3.20(m,2H),3.09–3.02(m,2H),2.85(t,J =7.5Hz,2H),2.37–2.28(m,1H),1.98–1.86(m,4H),1.78(s,AcOH,6H),1.72–1.42(m,8H),1.38–1.23(m,2H),0.75(dd,J=11.2,6.8Hz,6H).

[0316] Compound 19: Ala-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0317] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 70 H 120 N 22 O 20 m / z: 530.64499([M+3H]) 3+ ), 795.46224([M+2H] 2+ ), 1589.91350([M+H] + ).

[0318] 1 H NMR(600MHz,D2O)δ4.53–4.43(m,2H),4.33(dd,J=8.3,6.0Hz,1H),4.27(t,J=7.7Hz,3H),4.24–4.12 (m,6H),4.06–3.94(m,3H),3.77(q,J=7.6Hz,1H),3.73–3.67(m,2H),3.63–3.53(m,2H),3.51(q,J=7. 6Hz,1H),3.08(t,J=6.9Hz,2H),2.91–2.83(m,4H),2.29(t,J=7.6Hz,2H),2.26–2.12(m,7H),2.03–1 .46(m,AcOH,38H),1.39(d,J=7.1Hz,4H),1.36–1.30(m,2H),1.30–1.22(m,10H),0.92–0.77(m,12H).

[0319] Compound 20: Lys-Ala-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0320] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 71 H 125 N 23 O 18 m / z: 530.32804 ([M+3H]) 3+ ), 794.98683([M+2H] 2+ ), 1588.96546([M+H] + ).

[0321] 1 H NMR(600MHz,D2O)δ4.51(q,J=7.1Hz,1H),4.48–4.43(m,1H),4.31–4.25(m,3H),4.24–4.11(m,6H),4 .03(dd,J=8.4,4.9Hz,1H),3.98(d,J=7.6Hz,1H),3.87(t,J=6.7Hz,1H),3.79–3.67(m,3H),3.60–3. 47(m,3H),3.08(t,J=6.9Hz,2H),2.91–2.83(m,6H),2.28(t,J=7.6Hz,2H),2.21–2.13(m,5H),2.03– 1.64(m,AcOH,33H),1.64–1.47(m,11H),1.44–1.28(m,6H),1.28–1.21(m,12H),0.89–0.76(m,12H).

[0322] Compound 21: Lys-Glu-Ala-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0323] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 71 H 125 N 23 O 20 m / z: 540.99125([M+3H]) 3+ ), 810.98235([M+2H] 2+ ), 1620.94722([M+H] + ).

[0324] 1H NMR(600MHz,D2O)δ4.45(dd,J=9.4,4.9Hz,1H),4.30–4.24(m,3H),4.24–4.11(m,8H),4.02( dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.89(t,J=6.7Hz,1H),3.79–3.66(m,2H),3.60 –3.47(m,2H),3.08(t,J=6.9Hz,2H),2.91–2.83(m,6H),2.28(t,J=7.6Hz,2H),2.22–2.12(m ,6H),2.02–1.46(m,AcOH,41H),1.43–1.28(m,6H),1.28–1.20(m,12H),0.88–0.75(m,12H).

[0325] Compound 22: Lys-Glu-Pro-Val-Ala-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0326] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 71 H 125 N 23 O 20 m / z: 540.99088([M+3H]) 3+ ), 810.98168([M+2H] 2+ ), 1620.95237([M+H] + ).

[0327] 1H NMR (600MHz, D2O) δ4.51 (dd, J=9.6, 4.4Hz, 1H), 4.46 (dd, J=9.4, 4.8Hz, 1H), 4.35 (dd, J=8.4, 5.8Hz, 1H), 4.28 (dd, J= 8.3,6.1Hz,1H),4.25–4.13(m,7H),4.03(dd,J=8.3,4.9Hz,1H),3.98(d,J=7.6Hz,1H),3.94–3.88(m,2H),3.75–3.67( m,2H),3.63–3.56(m,1H),3.51(q,J=7.3Hz,1H),3.09(t,J=6.9Hz,2H),2.91–2.83(m,6H),2.29–2.21(m,4H),2.21–2 .13(m,4H),2.03–1.65(m,AcOH,31H),1.64–1.46(m,11H),1.45–1.28(m,6H),1.28–1.22(m,12H),0.89–0.78(m,12H).

[0328] Compound 23: Lys-Glu-Pro-Val-Pro-Ala-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0329] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 71 H 124 N 22 O 19 m / z: 398.24191([M+4H]) 4+ ), 530.65344([M+3H] 3+ ), 795.97791([M+2H] 2+ ).

[0330] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.6,4.4Hz,1H),4.44(dd,J=9.4,4.9Hz,1H),4.33(dd,J=8.4,5.8Hz,1H),4.31–4. 08(m,9H),4.03(dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.92–3.87(m,1H),3.79–3.73(m,1H),3.73–3.66(m, 2H),3.62–3.52(m,2H),3.51–3.46(m,1H),3.07(t,J=6.9Hz,2H),2.90–2.81(m,6H),2.29–2.21(m,2H),2.21–2.10 (m,5H),2.02–1.64(m,AcOH,37H),1.64–1.45(m,11H),1.44–1.27(m,6H),1.27–1.20(m,12H),0.91–0.74(m,12H).

[0331] Compound 24: Lys-Glu-Pro-Val-Pro-Gln-Ala-Ala-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0332] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 70 H 120 N 22 O 20 m / z: 398.23245([M+4H]) 4+ ), 530.64075([M+3H] 3+ ), 795.45752([M+2H] 2+ ).

[0333] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.7,4.4Hz,1H),4.45(q,J=7.1Hz,1H),4.33(dd,J=8.4,5.8Hz,1H),4.31–4.24(m ,3H),4.23–4.12(m,6H),4.03(dd,J=8.4,4.8Hz,1H),3.97(d,J=7.6Hz,1H),3.92–3.87(m,1H),3.79–3.73(m,1H) ,3.73–3.64(m,2H),3.62–3.48(m,3H),3.08(t,J=6.9Hz,2H),2.91–2.83(m,4H),2.32–2.21(m,4H),2.20–2.11( m,5H),2.03–1.71(m,AcOH,32H),1.71–1.45(m,10H),1.37–1.27(m,4H),1.27–1.17(m,12H),0.90–0.76(m,12H).

[0334] Compound 25: Lys-Glu-Pro-Ala-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0335] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 71 H 123 N 23 O 20 m / z: 405.48935([M+4H]) 4+ ), 540.31677([M+3H] 3+ ), 809.97144([M+2H] 2+ ).

[0336] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.5,4.4Hz,1H),4.48–4.41(m,2H),4.32–4.25(m,3H),4.21(dd,J=8.1,6 .7Hz,1H),4.19–4.12(m,5H),4.02(dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.92–3.85(m,1H),3.7 4–3.64(m,3H),3.60–3.46(m,3H),3.07(t,J=6.9Hz,2H),2.90–2.83(m,6H),2.30–2.20(m,4H),2.20–2. 12(m,5H),2.02–1.45(m,AcOH,48H),1.43–1.27(m,6H),1.27–1.21(m,12H),0.80(dd,J=6.8,4.4Hz,6H).

[0337] Compound 26: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Ala-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0338] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 71 H 125 N 23 O 20 m / z: 405.99350([M+4H]) 4+ ), 540.98883([M+3H] 3+ ), 810.97961([M+2H] 2+ ).

[0339] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.6,4.4Hz,1H),4.33(dd,J=8.4,5.8Hz,1H),4.31–4.23(m,2H ),4.23–4.09(m,8H),4.03(dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.93–3.86(m,1H),3. 81–3.66(m,2H),3.62–3.52(m,2H),3.07(t,J=6.9Hz,2H),2.86(m,6H),2.31–2.11(m,8H),2. 03–1.41(m,AcOH,48H),1.40–1.27(m,6H),1.25(dd,J=13.7,7.3Hz,12H),0.90–0.76(m,12H).

[0340] Compound 27: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Ala-Lys-Val-Ala-Ala-Gln (acetate)

[0341] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 70 H 120 N 20 O 20 m / z: 391.23177([M+4H]) 4+ ), 781.45496([M+2H] 2+ ).

[0342] 1 H NMR(600MHz,D2O)δ4.52–4.43(m,2H),4.33(dd,J=8.4,5.8Hz,1H),4.31–4.22(m,3H),4.21–4.10( m,6H),4.02(dd,J=8.5,4.8Hz,1H),3.96(d,J=7.7Hz,1H),3.92–3.84(m,1H),3.81–3.73(m,1H),3. 73–3.65(m,2H),3.61–3.52(m,2H),3.52–3.45(m,1H),2.91–2.81(m,6H),2.30–2.10(m,9H),2.02– 1.62(m,AcOH,31H),1.62–1.51(m,8H),1.41–1.27(m,6H),1.27–1.20(m,12H),0.90–0.75(m,12H).

[0343] Compound 28: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Ala-Val-Ala-Ala-Gln (acetate)

[0344] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 70 H 120 N 22 O 20 m / z: 398.23303([M+4H]) 4+ ), 530.64142([M+3H] 3+ ), 795.45850([M+2H] 2+ ).

[0345] 1 H NMR(600MHz,D2O)δ4.52–4.42(m,2H),4.33(dd,J=8.4,5.8Hz,1H),4.30–4.24(m,3H),4.23–4.09(m,6H), 4.02(dd,J=8.5,4.8Hz,1H),3.95(d,J=7.5Hz,1H),3.92–3.85(m,1H),3.79–3.73(m,1H),3.72–3.67(m,2 H),3.62–3.42(m,3H),3.08(t,J=6.9Hz,2H),2.87(t,J=7.6Hz,4H),2.30–2.25(m,2H),2.24–2.10(m,7H) ,2.02–1.65(m,AcOH,29H),1.64–1.48(m,8H),1.42–1.29(m,4H),1.28–1.19(m,12H),0.90–0.75(m,12H).

[0346] Compound 29: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Ala-Ala-Ala-Gln (acetate)

[0347] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 71 H 123 N 23 O 20 m / z: 405.48969([M+4H]) 4+ ), 540.31714([M+3H] 3+ ), 809.97205([M+2H] 2+ ).

[0348] 1 H NMR(600MHz,D2O)δ4.49(dd,J=9.6,4.3Hz,1H),4.45(dd,J=9.3,4.9Hz,1H),4.33(dd,J=8.4,5.8Hz ,1H),4.29–4.24(m,3H),4.19–4.13(m,7H),4.02(dd,J=8.4,4.8Hz,1H),3.89(t,J=6.8Hz,1H),3.7 9–3.66(m,3H),3.61–3.47(m,3H),3.08(t,J=6.9Hz,2H),2.86(q,J=8.0Hz,6H),2.28(t,J=7.6Hz,2 H),2.23–2.13(m,7H),2.02–1.46(m,AcOH,47H),1.42–1.18(m,20H),0.85(dd,J=14.2,6.7Hz,6H).

[0349] Compound 30: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Ala (acetate)

[0350] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 71 H 124 N 22 O 19 m / z: 398.24493([M+4H]) 4+ ), 530.65551([M+3H] 3+ ), 795.47815([M+2H] 2+ ), 1589.94194([M+H] + ).

[0351] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.6,4.4Hz,1H),4.46(dd,J=9.4,4.9Hz,1H),4.34(dd,J=8.4,5.8Hz,1H),4.3 1–4.24(m,3H),4.24–4.20(m,1H),4.20–4.09(m,5H),4.02–3.96(m,2H),3.93–3.86(m,1H),3.80–3.74(m,1H ),3.74–3.67(m,2H),3.63–3.43(m,3H),3.08(t,J=6.9Hz,2H),2.91–2.82(m,6H),2.31–2.12(m,7H),2.01–1 .65(m,AcOH,34H),1.64–1.46(m,11H),1.35(dd,J=16.0,8.1Hz,6H),1.27–1.20(m,12H),0.92–0.77(m,12H).

[0352] Compound 31: Lys-Glu-Gly-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0353] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 70 H 123 N 23 O 20 m / z: 402.49240([M+4H]) 4+ ), 536.31955([M+3H] 3+ ), 803.97442([M+2H] 2+ ), 1606.91471([M+H] + ).

[0354] 1H NMR (600MHz, D2O) δ4.45 (dd, J=9.4, 4.9Hz, 1H), 4.32 (d, J=8.1Hz, 1H), 4.30–4.25 (m, 2H), 4.24–4.19 (m, 2H), 4.19–4. 11(m,5H),4.02(dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.91–3.84(m,2H),3.82–3.73(m,2H),3.69(q,J=7.7,7. 0Hz,1H),3.59–3.52(m,1H),3.52–3.46(m,1H),3.07(t,J=6.9Hz,2H),2.90–2.82(m,6H),2.28(t,J=7.6Hz,2H),2.23– 2.12(m,6H),2.02–1.64(m,AcOH,35H),1.63–1.45(m,11H),1.44–1.27(m,6H),1.27–1.21(m,9H),0.88–0.73(m,12H).

[0355] Compound 32: Lys-Glu-Pro-Val-Gly-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0356] High-resolution mass spectrometry (TOF-HRMS), molecular formula: C 70 H 123 N 23 O 20 m / z: 402.49278([M+4H]) 4+ ), 536.31975([M+3H] 3+ ), 803.97422([M+2H] 2+ ), 1606.92969([M+H] + ).

[0357] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.7,4.4Hz,1H),4.46(dd,J=9.4,4.9Hz,1H),4.36(dd,J=8.3,5.9Hz,1H),4.28(dd,J=8.3 ,6.1Hz,1H),4.24–4.14(m,6H),4.03(dd,J=8.3,4.9Hz,1H),3.97(dd,J=10.0,7.5Hz,2H),3.90(t,J=6.8Hz,1H),3.87–3. 78(m,2H),3.76–3.67(m,2H),3.61–3.54(m,1H),3.54–3.47(m,1H),3.08(t,J=6.9Hz,2H),2.91–2.83(m,6H),2.26–2.13( m,8H),2.03–1.87(m,9H),1.87–1.45(m,AcOH,33H),1.44–1.28(m,6H),1.26(dd,J=18.4,7.2Hz,9H),0.89–0.79(m,12H).

[0358] Compound 33: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0359] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 70 H 123 N 23 O 20 m / z: 402.48917([M+4H]) 4+ ), 536.31647([M+3H] 3+ ), 803.97101([M+2H] 2+ ).

[0360] 1H NMR(600MHz,D2O)δ4.51(dd,J=9.6,4.4Hz,1H),4.34(dd,J=8.4,5.8Hz,1H),4.31–4.11(m,9 H),4.03(dd,J=8.4,4.9Hz,1H),3.97(d,J=7.6Hz,1H),3.92–3.86(m,1H),3.85–3.74(m,3H) ,3.74–3.66(m,1H),3.62–3.52(m,2H),3.06(t,J=7.0Hz,2H),2.91–2.83(m,6H),2.31–2.11 (m,8H),2.03–1.43(m,AcOH,45H),1.40–1.27(m,6H),1.27–1.23(m,9H),0.91–0.77(m,12H).

[0361] Compound 34: Lys-Glu-Pro-Val-Gly-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0362] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 67 H 119 N 23 O 20 m / z: 392.48178([M+4H]) 4+ ), 522.97314([M+3H] 3+ ), 784.45752([M+2H] 2+ ).

[0363] 1 H NMR(600MHz,D2O)δ4.51(dd,J=9.6,4.4Hz,1H),4.35(dd,J=8.3,5.9Hz,1H),4.25–4.12(m,7H),4.03(d d,J=8.4,4.8Hz,1H),3.96(dd,J=10.6,7.5Hz,2H),3.92–3.86(m,1H),3.85–3.77(m,4H),3.75–3.68(m, 1H),3.60–3.53(m,1H),3.06(t,J=7.0Hz,2H),2.92–2.83(m,6H),2.28–2.12(m,7H),2.05–1.41(m,AcOH ,42H),1.40–1.27(m,6H),1.27–1.24(m,9H),0.85(dd,J=11.0,6.7Hz,6H),0.80(dd,J=6.8,5.1Hz,6H).

[0364] Compound 35: Lys-Glu-Gly-Val-Gly-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0365] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 64 H 115 N 23 O 20 m / z: 382.47363([M+4H]) 4+ ), 509.62903([M+3H] 3+ ), 763.93982([M+2H] 2+ ).

[0366] 1 H NMR (600MHz, D2O) δ4.27–4.13(m,8H),4.05–4.00(m,2H),3.97(d,J=7.6Hz,1H),3.93–3.87(m,2H),3.85–3.76(m,5H),3.06(t,J=7. 0Hz,2H),2.90–2.83(m,6H),2.24–2.12(m,6H),2.02–1.41(m,AcOH,40H),1.39–1.27(m,6H),1.27–1.22(m,9H),0.85–0.76(m,12H).

[0367] Compound 36: Lys-Glu-Gly-Val-Pro-Gln-Ala-Lys-Gly-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0368] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 67 H 119 N 23 O 20 m / z: 392.48172([M+4H]) 4+ ), 522.97314([M+3H] 3+ ), 783.95605([M+2H] 2+ ).

[0369] 1H NMR(600MHz,D2O)δ4.32(d,J=8.0Hz,1H),4.28(dd,J=8.2,6.7Hz,1H),4.25–4.12(m,8H),4 .03(dd,J=8.4,4.8Hz,1H),3.97(d,J=7.6Hz,1H),3.92–3.83(m,2H),3.83–3.73(m,4H),3.6 0–3.53(m,1H),3.06(t,J=7.0Hz,2H),2.91–2.83(m,6H),2.28(t,J=7.5Hz,2H),2.24–2.12( m,5H),2.02–1.42(m,AcOH,43H),1.41–1.27(m,6H),1.27–1.22(m,9H),0.91–0.72(m,12H).

[0370] Compound 37: Lys-Glu-Gly-Val-Gly-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0371] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 67 H 119 N 23 O 20 m / z: 392.48172([M+4H]) 4+ ), 522.97314([M+3H] 3+ ), 783.95605([M+2H] 2+ ).

[0372] 1 H NMR(600MHz,D2O)δ4.44(dd,J=9.4,4.9Hz,1H),4.29–4.12(m,8H),4.04–3.99(m, 2H),3.96(d,J=7.6Hz,1H),3.91–3.82(m,3H),3.81(s,2H),3.72–3.65(m,1H),3. 52–3.44(m,1H),3.06(t,J=6.9Hz,2H),2.89–2.81(m,6H),2.23–2.12(m,7H),2.0 1–1.45(m,AcOH,43H),1.42–1.26(m,6H),1.26–1.20(m,9H),0.83–0.77(m,12H).

[0373] Compound 38: Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys (acetate)

[0374] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 51 H 88 N 16 O 14 m / z: 383.89569([M+3H]) 3+ ), 575.33978([M+2H] 2+ ).

[0375] 1 H NMR(600MHz,D2O)δ4.45(dd,J=9.2,5.0Hz,1H),4.39(dd,J=8.3,6.4Hz,1H),4.29–4.21( m,4H),4.19–4.12(m,3H),4.01(dd,J=7.8,5.5Hz,1H),3.78–3.73(m,1H),3.72–3.67(m,1 H),3.61–3.48(m,4H),3.08(t,J=6.8Hz,2H),2.88–2.83(m,4H),2.30–2.26(m,4H),2.20 –2.13(m,3H),2.08–1.46(m,AcOH,38H),1.39–1.22(m,7H),0.86(dd,J=10.3,6.7Hz,6H).

[0376] Compound 39: Pro-Val-Pro-Gln-Ala (acetate)

[0377] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 23 H 38 N6O7; m / z: 511.2871 ([M+H]) + ).

[0378] 1 H NMR(600MHz,D2O)δ4.41(d,J=7.3Hz,1H),4.38–4.32(m,2H),4.20(dd,J=8.7,5.9 Hz,1H),4.11–4.04(m,1H),3.85–3.77(m,1H),3.62(dt,J=10.3,7.2Hz,1H),3.39– 3.29(m,2H),2.39–2.31(m,3H),2.28–2.20(m,1H),2.08–1.90(m,AcOH,10H),1.86 –1.80(m,1H),1.26(d,J=7.3Hz,3H),0.94(d,J=6.8Hz,3H),0.88(d,J=6.8Hz,3H).

[0379] Compound 40: Pro-Val-Pro-Glu-Ala (acetate)

[0380] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 23 H 37 N5O8; m / z: 512.27081 ([M+H]) + ).

[0381] 1 H NMR(600MHz,D2O)δ4.41(d,J=7.3Hz,1H),4.38–4.32(m,2H),4.23(dd,J=8.9 ,5.6Hz,1H),4.13–4.09(m,1H),3.86–3.76(m,1H),3.62(dt,J=10.2,7.2Hz,1 H),3.38–3.28(m,2H),2.43–2.35(m,3H),2.26–2.16(m,1H),2.12–1.72(m,Ac OH,12H),1.28(d,J=7.2Hz,3H),0.94(d,J=6.8Hz,3H),0.87(d,J=6.8Hz,3H).

[0382] Compound 41: Pro-Val-Pro-Ile-Ala (acetate)

[0383] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 24 H 41 N5O6; m / z: 496.31158 ([M+H]) + ).

[0384] 1H NMR(600MHz,D2O)δ4.36–4.27(m,3H),4.03(q,J=7.2Hz,1H),3.98(d,J=7.8Hz,1H),3.79–3.72(m ,1H),3.60–3.53(m,1H),3.34–3.21(m,2H),2.36–2.25(m,1H),2.21–2.12(m,1H),2.03–1.96(m, 1H),1.96–1.81(m,AcOH,7H),1.80–1.69(m,2H),1.43–1.33(m,1H),1.20(d,J=7.2Hz,3H),1.13– 1.02(m,1H),0.88(d,J=6.8Hz,3H),0.85–0.80(m,6H),0.80–0.77(m,1H),0.74(t,J=7.4Hz,3H).

[0385] Compound 42: Pro-Val-Pro-Ala

[0386] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 18 H 30 N4O5; m / z: 383.22791 ([M+H]) + ).

[0387] 1 H NMR(600MHz,D2O)δ4.36(d,J=7.2Hz,1H),4.30(dd,J=8.5,6.3Hz,1H),4.24(d d,J=8.1,6.5Hz,1H),4.00(q,J=7.2Hz,1H),3.79–3.72(m,1H),3.60–3.52(m, 1H),3.31–3.23(m,2H),2.33–2.26(m,1H),2.20–2.13(m,1H),2.09–1.69(m,A cOH,10H),1.21(d,J=7.3Hz,3H),0.88(d,J=6.9Hz,3H),0.81(d,J=6.8Hz,3H).

[0388] Compound 43: Glu-Pro-Val-Pro-Gln-Ala (acetate)

[0389] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 28 H 45 N7O 10 m / z: 640.32892 ([M+H]) + ).

[0390] 1 H NMR(600MHz,D2O)δ4.34(dd,J=8.3,6.5Hz,1H),4.25–4.15(m,3H),4.08(dd,J =8.8,5.9Hz,1H),4.04–4.00(m,1H),3.73–3.65(m,1H),3.57–3.41(m,3H),2. 35(t,J=7.3Hz,2H),2.24–2.20(m,2H),2.16–2.07(m,2H),2.04–1.75(m,AcOH ,11H),1.73–1.63(m,2H),1.17(d,J=7.2Hz,3H),0.80(dd,J=12.9,6.8Hz,6H).

[0391] Compound 44: Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0392] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 29 H 50 N8O8; m / z: 320.19421 ([M+2H]) 2+ ).

[0393] 1 H NMR(600MHz,D2O)δ4.29–4.19(m,3H),4.13–4.06(m,2H),3.95(dd,J=8.2,5.1 Hz,1H),3.73–3.68(m,1H),3.52–3.47(m,1H),3.24–3.16(m,2H),2.79(t,J=7. 5Hz,2H),2.25–2.19(m,3H),2.14–2.08(m,1H),1.97–1.57(m,AcOH,17H),1.53 –1.45(m,3H),1.21–1.17(m,5H),0.82(d,J=6.8Hz,3H),0.75(d,J=6.7Hz,3H).

[0394] Compound 45: Glu-Pro-Val-Pro-Gln-Ala-Arg (acetate)

[0395] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N 11 O 11 m / z: 398.71835([M+2H]) 2+ ), 796.42944([M+H]+ ).

[0396] 1 H NMR(600MHz,D2O)δ4.38(dd,J=8.3,6.4Hz,1H),4.27–4.19(m,3H),4.19–4.10(m,2H),4.03(d d,J=8.3,4.9Hz,1H),3.78–3.70(m,1H),3.60–3.46(m,3H),3.07–3.02(m,2H),2.30(t,J=7.2 Hz,2H),2.25(t,J=7.6Hz,2H),2.19–2.11(m,2H),2.06–1.78(m,AcOH,12H),1.76–1.67(m,3H ),1.60–1.53(m,1H),1.48–1.42(m,2H),1.24(d,J=7.1Hz,3H),0.85(dd,J=12.8,6.7Hz,6H).

[0397] Compound 46: Ala-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0398] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 32 H 55 N9O9; m / z: 355.71259 ([M+2H]) 2+ ), 710.41809([M+H] + ).

[0399] 1H NMR (600MHz, DMSO-d6) δ8.23(d,J=7.7Hz,1H),8.08(d,J=8.1Hz,1H),7.87(d,J=8.4Hz,1H),7.68(s,1H),7.38(d,J=6.4Hz,1H),6.75(s, 1H),4.40–4.37(m,1H),4.33–4.25(m,3H),4.15(t,J=7.2Hz,3H),3.78(d,J=6.4Hz,1H),3.66(dd,J=32.4,8.0Hz,2H),3.58–3.51(m,2H), 3.43(d,J=7.0Hz,1H),3.36–3.27(m,1H),2.67(t,J=7.7Hz,3H),2.19–2.07(m,4H),2.01–1.69(m,AcOH,24H),1.63–1.60(m,1H),1.55–1. 41(m,5H),1.26(d,1H),1.18(d,J=7.2Hz,6H),1.11(d,J=6.7Hz,3H),1.04(d,J=6.6Hz,1H),0.87(d,J=6.7Hz,3H),0.83(d,J=6.9Hz,3H).

[0400] Compound 47: Glu-Pro-Val-Pro-Ala-Ala-Lys (acetate)

[0401] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 32 H 54 N8O 10 m / z: 356.20514([M+2H]) 2+ ), 711.40198([M+H] + ).

[0402] 1 H NMR(600MHz,D2O)δ4.45(dd,J=8.4,6.4Hz,1H),4.33–4.27(m,3H),4.21(dd,J=1 5.1,7.2Hz,2H),4.07(dd,J=8.1,5.1Hz,1H),3.86–3.75(m,1H),3.67–3.54(m,3H ),2.91(t,J=7.5Hz,2H),2.37(t,J=6.9Hz,2H),2.26–2.18(m,2H),2.14–1.69(m ,AcOH,13H),1.65–1.58(m,3H),1.35–1.29(m,9H),0.92(dd,J=13.6,6.8Hz,6H).

[0403] Compound 48: Glu-Pro-Val-Pro-Gln-Ala-Ala (acetate)

[0404] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 50 N8O 11 m / z: 356.18692([M+2H]) 2+ ), 711.36597([M+H] + ).

[0405] 1 H NMR(600MHz,D2O)δ4.34(dd,J=8.3,6.5Hz,1H),4.24–4.16(m,3H),4.12–4.0 6(m,2H),4.03–3.99(m,1H),3.73–3.65(m,1H),3.56–3.41(m,3H),2.35(t,J= 7.3Hz,2H),2.22–2.18(m,2H),2.15–2.07(m,2H),2.04–1.73(m,AcOH,11H),1 .71–1.63(m,2H),1.18(dd,J=11.2,7.2Hz,6H),0.80(dd,J=13.0,6.8Hz,6H).

[0406] Compound 49: Asp-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0407] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 33 H 55 N9O 11 m / z: 377.70767([M+2H]) 2+ ), 754.40814([M+H] + ).

[0408] 1H NMR(600MHz,D2O)δ4.36–4.29(m,2H),4.23–4.14(m,2H),4.13–4.06(m,2H),3.97(dd,J=8.3,5.0Hz,1H ),3.74–3.65(m,1H),3.56–3.46(m,2H),3.44–3.39(m,1H),2.79(t,J=7.5Hz,2H),2.65(dd,J=17.5,3. 3Hz,1H),2.38(dd,J=17.5,10.7Hz,1H),2.21(t,J=7.6Hz,2H),2.15–2.06(m,2H),1.93–1.75(m,AcOH, 9H),1.72–1.61(m,3H),1.54–1.45(m,3H),1.21(dd,J=13.1,7.5Hz,5H),0.80(dd,J=11.0,6.7Hz,6H).

[0409] Compound 50: Asn-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0410] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 33 H 56 N 10 O 10 m / z: 377.21542([M+2H]) 2+ ), 753.42365([M+H] + ).

[0411] 1 H NMR(600MHz,D2O)δ4.55(dd,J=9.3,4.0Hz,1H),4.47(dd,J=8.4,6.1Hz,1H),4.34–4.29(m, 2H),4.26–4.19(m,2H),4.07(dd,J=8.1,5.1Hz,1H),3.83–3.77(m,1H),3.69–3.53(m,3H),2 .94–2.90(m,3H),2.72(dd,J=17.0,9.3Hz,1H),2.33(t,J=7.6Hz,2H),2.27–2.19(m,2H),2. 05–1.71(m,AcOH,15H),1.65–1.57(m,3H),1.34–1.30(m,5H),0.91(dd,J=13.4,6.7Hz,6H).

[0412] Compound 51: Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala (acetate)

[0413] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 59 H 102 N 18 O 16 m / z: 440.59735([M+3H]) 3+ ), 660.39252([M+2H] 2+ ).

[0414] 1 H NMR(600MHz,D2O)δ4.38(dd,J=9.3,4.8Hz,1H),4.33(dd,J=8.3,6.5Hz,1H),4.22–4.12(m,5H ),4.10–4.04(m,3H),3.93–3.88(m,2H),3.75–3.67(m,1H),3.66–3.58(m,1H),3.56–3.41(m,4 H),3.00(t,J=6.9Hz,2H),2.82–2.76(m,4H),2.24–2.19(m,4H),2.14–2.06(m,3H),1.97–1.41 (m,AcOH,37H),1.33–1.12(m,10H),0.80(dd,J=9.7,6.7Hz,6H),0.74(dd,J=11.0,6.8Hz,6H).

[0415] Compound 52: Glu-Pro-Ala-Pro-Gln-Ala-Lys (acetate)

[0416] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 32 H 53 N9O 11 m / z: 370.6998([M+2H]) 2+ ), 740.39221([M+H] + ).

[0417] 1H NMR(600MHz,D2O)δ4.43(q,J=7.1Hz,1H),4.33(dd,J=8.4,6.5Hz,1H),4.26(dd,J=8.4,5.8Hz,1H),4.22(d d,J=7.6,4.6Hz,1H),4.18–4.11(m,2H),4.01(dd,J=8.3,5.0Hz,1H),3.67(dt,J=10.0,6.7Hz,1H),3.61–3 .54(m,1H),3.53–3.44(m,2H),2.84(t,J=7.5Hz,2H),2.30(t,J=7.2Hz,2H),2.25(t,J=7.6Hz,2H),2.21–2 .12(m,2H),2.08–1.77(m,AcOH,11H),1.77–1.63(m,3H),1.60–1.49(m,3H),1.24(dd,J=10.4,7.1Hz,6H).

[0418] Compound 53: Glu-Ala-Val-Pro-Gln-Ala-Lys (acetate)

[0419] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 32 H 55 N9O 11 m / z: 371.7081([M+2H]) 2+ ), 742.4089([M+H] + ).

[0420] 1 H NMR(600MHz,D2O)δ4.23–4.18(m,3H),4.13–4.06(m,2H),3.95(dd,J=8.3,5.0Hz,1 H),3.85–3.82(m,1H),3.73–3.66(m,1H),3.53–3.47(m,1H),2.79(t,J=7.5Hz,2H), 2.22–2.17(m,4H),2.13–2.07(m,1H),1.99–1.70(m,AcOH,9H),1.69–1.59(m,2H), 1.53–1.45(m,3H),1.26–1.12(m,9H),0.79(d,J=6.7Hz,3H),0.76(d,J=6.7Hz,3H).

[0421] Compound 54: Glu-Pro-Val-Ala-Gln-Ala-Lys (acetate)

[0422] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 32 H 55 N9O 11 m / z: 371.70767([M+2H]) 2+ ), 742.40802([M+H] + ).

[0423] 1 H NMR(600MHz,DMSO-d6+D2O)δ4.34(dd,J=8.3,6.0Hz,1H),4.15–4.05(m,4H), 3.97–3.87(m,2H),3.51(d,J=9.8Hz,1H),3.46–3.38(m,1H),2.74(t,J=7.5Hz ,2H),2.23(t,J=7.1Hz,2H),2.17–2.06(m,3H),1.95–1.67(m,AcOH,10H),1. 65–1.59(m,1H),1.53–1.42(m,3H),1.22–1.13(m,8H),0.78(d,J=6.8Hz,6H).

[0424] Compound 55: Glu-Pro-Val-Pro-Glu-Ala-Lys (acetate)

[0425] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 56 N8O 12 m / z: 385.2083([M+2H]) 2+ ), 769.4092([M+H] + ).

[0426] 1H NMR(600MHz,D2O)δ4.33(dd,J=8.3,6.5Hz,1H),4.21–4.16(m,3H),4.11(t,J=7.1Hz,1H),4. 07(dd,J=8.8,5.7Hz,1H),3.98(dd,J=8.5,4.8Hz,1H),3.71–3.66(m,1H),3.55–3.41(m,3H), 2.78(t,J=7.5Hz,2H),2.36–2.14(m,5H),2.13–2.05(m,2H),1.99–1.74(m,AcOH,11H),1.70 –1.61(m,3H),1.53–1.45(m,3H),1.20(dd,J=15.4,7.5Hz,5H),0.80(dd,J=12.5,6.7Hz,6H).

[0427] Compound 56: Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val (acetate)

[0428] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 56 H 97 N 17 O 15 m / z: 416.91827([M+3H]) 3+ ), 624.87354([M+2H] 2+ ).

[0429] 1 H NMR(600MHz,D2O)δ4.43(dd,J=9.3,4.9Hz,1H),4.38(dd,J=8.4,6.4Hz,1H),4.27–4.19(m,5H),4 .16–4.10(m,3H),3.88(d,J=6.2Hz,1H),3.74(dt,J=10.1,6.9Hz,1H),3.71–3.63(m,1H),3.60–3 .46(m,4H),3.07–3.02(m,2H),2.87–2.82(m,4H),2.29–2.24(m,4H),2.19–2.11(m,3H),2.04–1. 43(m,AcOH,37H),1.37–1.20(m,7H),0.85(dd,J=10.1,6.8Hz,6H),0.74(dd,J=11.8,6.9Hz,6H).

[0430] Compound 57: Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (acetate)

[0431] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 45 H 76 N 14 O 13 m / z: 511.29254([M+2H]) 2+ ).

[0432] 1 H NMR(600MHz,D2O)δ4.39(dd,J=8.6,5.5Hz,1H),4.32(dd,J=8.3,6.5Hz,1H),4.22–4.13(m,4H),4.10–4.0 3(m,2H),3.93(dd,J=7.9,5.2Hz,1H),3.72–3.60(m,2H),3.54–3.40(m,4H),3.00(t,J=6.9Hz,2H),2.80( t,J=7.5Hz,2H),2.24–2.18(m,4H),2.14–2.06(m,3H),1.98–1.70(m,AcOH,19H),1.69–1.60(m,4H),1.55 –1.47(m,4H),1.45–1.39(m,2H),1.34–1.24(m,2H),1.16(d,J=7.1Hz,3H),0.80(dd,J=10.4,6.7Hz,6H).

[0433] Compound 58: Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro (acetate)

[0434] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 39 H 64 N 10 O 12 m / z: 433.24188([M+2H]) 2+ ), 865.47662([M+H] + ).

[0435] 1H NMR(600MHz,D2O)δ4.41(dd,J=8.5,5.5Hz,1H),4.33(dd,J=8.3,6.5Hz,1H),4 .22–4.14(m,3H),4.11–4.02(m,3H),3.75–3.67(m,1H),3.59–3.41(m,5H),2. 81(t,J=7.4Hz,2H),2.25–2.19(m,4H),2.15–1.61(m,AcOH,26H),1.57–1.48( m,3H),1.37–1.23(m,2H),1.16(d,J=7.0Hz,3H),0.80(dd,J=11.5,6.7Hz,6H).

[0436] Compound 59: Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg (acetate)

[0437] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 40 H 69 N 13 O 10 m / z: 298.18314([M+3H]) 3+ ), 446.77118([M+2H] 2+ ), 892.53497([M+H] + ).

[0438] 1 H NMR(600MHz,D2O)δ4.40(dd,J=8.6,5.5Hz,1H),4.30–4.19(m,4H),4.11–4.05(m,2H),3.93(dd,J =7.9,5.2Hz,1H),3.73–3.63(m,2H),3.52–3.45(m,2H),3.26–3.17(m,2H),3.01(t,J=6.9Hz,2H) ,2.81(t,J=7.5Hz,2H),2.26–2.19(m,3H),2.14–2.09(m,2H),1.94–1.62(m,AcOH,24H),1.57–1. 40(m,7H),1.34–1.26(m,2H),1.16(d,J=7.3Hz,3H),0.82(d,J=6.8Hz,3H),0.76(d,J=6.7Hz,3H).

[0439] Compound 60: Pro-Val-Pro-Gln-Ala-Lys-Pro (acetate)

[0440] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N9O9; m / z: 368.7207 ([M+2H]) 2+ ), 736.4343([M+H] + ).

[0441] 1 H NMR(600MHz,D2O)δ4.46(dd,J=8.4,5.6Hz,1H),4.34(d,J=7.4Hz,1H),4.29(dd,J=8.5,6.2Hz,1H),4.27– 4.24(m,1H),4.16–4.07(m,3H),3.78–3.70(m,1H),3.66–3.60(m,1H),3.58–3.45(m,2H),3.30–3.22(m,2H ),2.86(t,J=7.5Hz,2H),2.32–2.24(m,3H),2.19–2.13(m,1H),2.12–2.03(m,1H),2.02–1.69(m,AcOH,19 H),1.61–1.52(m,3H),1.42–1.27(m,2H),1.23–1.20(m,3H),0.87(d,J=6.8Hz,3H),0.81(d,J=6.7Hz,3H).

[0442] Compound 61: D-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0443] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4978 ([M+4H]) 4+ ), 549.6611([M+3H] 3+ ), 823.9882([M+2H] 2+ ).

[0444] 1H NMR(600MHz,D2O)δ4.46–4.35(m,2H),4.27(dd,J=8.3,6.1Hz,1H),4.22–4.04(m,9H ),3.96(dd,J=8.4,4.8Hz,1H),3.90(d,J=7.7Hz,1H),3.84(t,J=6.6Hz,1H),3.74–3. 59(m,3H),3.56–3.38(m,3H),3.01(t,J=6.9Hz,2H),2.83–2.76(m,6H),2.23–2.06( m,9H),1.96–1.39(m,AcOH,48H),1.18(dd,J=22.0,7.2Hz,15H),0.83–0.67(m,12H).

[0445] Compound 62: Lys-D-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0446] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4976([M+4H]) 4+ ), 549.6608([M+3H] 3+ ).

[0447] 1 H NMR(600MHz,D2O)δ4.50(dd,J=9.8,4.4Hz,1H),4.37(dd,J=9.4,4.9Hz,1H),4.25–4.03(m,10H),3.95(d d,J=8.5,4.8Hz,1H),3.89(d,J=7.7Hz,1H),3.83(t,J=6.6Hz,1H),3.74–3.68(m,1H),3.64–3.55(m,2H) ,3.52–3.38(m,2H),3.00(t,J=6.9Hz,2H),2.83–2.75(m,6H),2.20(t,J=7.6Hz,2H),2.12–2.06(m,6H), 1.92–1.41(m,AcOH,47H), 1.32–1.13(m,15H), 0.78(dd,J=9.7,6.7Hz,6H), 0.73(dd,J=6.8,4.1Hz,6H).

[0448] Compound 63: Lys-Glu-D-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0449] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4976([M+4H]) 4+ ), 549.6609([M+3H] 3+ ), 823.9878([M+2H] 2+ ).

[0450] 1 H NMR(600MHz,D2O)δ4.45(dd,J=9.4,4.4Hz,1H),4.37(dd,J=9.4,4.9Hz,1H),4.29–4.25(m,1H ),4.21–4.04(m,8H),3.96–3.94(m,1H),3.89(d,J=7.7Hz,1H),3.79(d,J=6.9Hz,1H),3.68–3. 59(m,2H),3.54–3.35(m,3H),3.00(t,J=6.9Hz,2H),2.82–2.74(m,6H),2.23–2.19(m,2H),2. 15–2.02(m,6H),1.95–1.41(m,AcOH,48H),1.17(dd,J=21.6,7.2Hz,15H),0.83–0.64(m,12H).

[0451] Compound 64: Lys-Glu-Pro-D-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0452] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4978 ([M+4H]) 4+ ), 549.6609([M+3H] 3+ ), 823.9878([M+2H] 2+ ).

[0453] 1H NMR(600MHz,D2O)δ4.46(dd,J=10.2,3.9Hz,1H),4.41–4.31(m,2H),4.27–4.18(m,3H), 4.15–4.02(m,7H),3.95–3.92(m,1H),3.89(d,J=7.7Hz,1H),3.83(t,J=6.6Hz,1H),3.6 7–3.50(m,5H),3.47–3.25(m,2H),3.00(t,J=6.9Hz,2H),2.81–2.76(m,6H),2.23–2.06 (m,10H),1.94–1.41(m,AcOH,50H),1.17(dd,J=19.9,7.1Hz,17H),0.76–0.68(m,12H).

[0454] Compound 65: Lys-Glu-Pro-Val-D-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0455] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4979([M+4H]) 4+ ), 549.6612([M+3H] 3+ ), 823.9883([M+2H] 2+ ).

[0456] 1 H NMR(600MHz,D2O)δ4.42(dd,J=9.9,4.2Hz,1H),4.34(dd,J=9.2,5.2Hz,1H),4.31–4.23(m,2H) ,4.22–4.18(m,2H),4.15–4.05(m,6H),3.94(dd,J=8.4,4.8Hz,1H),3.89(d,J=7.7Hz,1H),3.8 2(t,J=6.7Hz,1H),3.65(d,J=6.8Hz,3H),3.57–3.36(m,3H),3.00(t,J=6.9Hz,2H),2.82–2.75 (m,6H),2.18–2.03(m,9H),1.95–1.40(m,AcOH,46H),1.37–1.08(m,16H),0.80–0.68(m,12H).

[0457] Compound 66: Lys-Glu-Pro-Val-Pro-D-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0458] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4978 ([M+4H]) 4+ ), 549.6611([M+3H] 3+ ), 823.9881([M+2H] 2+ ).

[0459] 1 H NMR(600MHz,D2O)δ4.44–4.35(m,2H),4.26–4.04(m,10H),3.94(dd,J=8.5,4.8Hz,1H),3. 89(d,J=7.7Hz,1H),3.82(t,J=6.8Hz,1H),3.76–3.68(m,1H),3.64(d,J=8.7Hz,2H),3.55 –3.37(m,3H),3.00(t,J=6.9Hz,2H),2.83–2.75(m,6H),2.19–2.05(m,9H),2.02–1.38(m, AcOH,47H),1.34–1.15(m,15H),0.78(dd,J=11.2,6.7Hz,6H),0.73(dd,J=6.7,3.9Hz,6H).

[0460] Compound 67: Lys-Glu-Pro-Val-Pro-Gln-D-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0461] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4961([M+4H]) 4+ ), 549.6615([M+3H] 3+ ), 823.9886([M+2H] 2+ ).

[0462] 1H NMR(600MHz,D2O)δ4.47–4.38(m,2H),4.26(dd,J=8.4,5.8Hz,1H),4.22–4.04(m,9H),3.95 (dd,J=8.5,4.8Hz,1H),3.89(d,J=7.7Hz,1H),3.82(t,J=6.8Hz,1H),3.74–3.56(m,3H),3. 54–3.35(m,3H),3.00(t,J=6.9Hz,2H),2.86–2.74(m,6H),2.23–2.05(m,9H),1.94–1.38(m ,AcOH,46H),1.29–1.14(m,15H),0.78(dd,J=10.5,6.7Hz,6H),0.73(dd,J=6.8,3.9Hz,6H).

[0463] Compound 68: Lys-Glu-Pro-Val-Pro-Gln-Ala-D-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0464] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4977([M+4H]) 4+ ), 549.6615([M+3H] 3+ ), 823.9879([M+2H] 2+ ).

[0465] 1 H NMR(600MHz,D2O)δ4.47–4.40(m,2H),4.26(dd,J=8.4,5.9Hz,1H),4.23–4.15(m,3H),4.14 –4.01(m,6H),3.95(dd,J=8.5,4.8Hz,1H),3.89(d,J=7.7Hz,1H),3.82(t,J=6.8Hz,1H),3. 71–3.44(m,5H),3.01(t,J=6.9Hz,2H),2.82–2.73(m,6H),2.23–2.05(m,9H),1.93–1.40(m ,AcOH,46H),1.30–1.14(m,15H),0.79(dd,J=12.5,6.7Hz,6H),0.73(dd,J=6.8,4.6Hz,6H).

[0466] Compound 69: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-D-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0467] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4976([M+4H]) 4+ ), 549.6608([M+3H] 3+ ).

[0468] 1 H NMR(600MHz,D2O)δ4.42(dt,J=8.2,4.3Hz,2H),4.26(dd,J=8.4,5.9Hz,1H),4.23–4.14(m,3H) ,4.14–4.05(m,6H),3.94(dd,J=8.5,4.8Hz,1H),3.89(d,J=7.7Hz,1H),3.81(q,J=7.7,7.2Hz, 1H),3.74–3.58(m,3H),3.55–3.38(m,3H),2.98(t,J=7.0Hz,2H),2.82–2.75(m,6H),2.23–2.0 5(m,9H),1.93–1.13(m,AcOH,62H),0.79(dd,J=13.2,6.7Hz,6H),0.73(dd,J=6.8,4.7Hz,6H).

[0469] Compound 70: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-D-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0470] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4978 ([M+4H]) 4+ ), 549.6611([M+3H] 3+ ), 823.9883([M+2H] 2+ ).

[0471] 1H NMR(600MHz,D2O)δ4.45–4.36(m,2H),4.27–4.14(m,6H),4.12–4.05(m,4H),3.95(dd, J=8.4,4.8Hz,1H),3.90(d,J=7.7Hz,1H),3.82(t,J=6.8Hz,1H),3.66(dd,J=35.9,8.3 Hz,3H),3.55–3.39(m,3H),2.99(t,J=7.0Hz,2H),2.82–2.75(m,6H),2.23–2.07(m,9H ),1.93–1.15(m,AcOH,63H),0.79(dd,J=13.9,6.7Hz,6H),0.74(dd,J=6.8,2.0Hz,6H).

[0472] Compound 71: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-D-Lys-Val-Ala-Ala-Gln (acetate)

[0473] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4976([M+4H]) 4+ ), 549.6609([M+3H] 3+ ), 823.9878([M+2H] 2+ ).

[0474] 1 H NMR(600MHz,D2O)δ4.45–4.36(m,2H),4.26(dd,J=8.4,5.8Hz,1H),4.22–4.16(m,3H),4.12– 4.05(m,6H),3.96–3.91(m,2H),3.82(t,J=6.8Hz,1H),3.73–3.58(m,3H),3.54–3.36(m,3H) ,3.00(t,J=7.0Hz,2H),2.82–2.75(m,6H),2.22–2.06(m,9H),1.93–1.41(m,AcOH,45H),1.3 2–1.14(m,15H),0.79(dd,J=13.8,6.7Hz,6H),0.74(d,J=6.8Hz,3H),0.70(d,J=6.8Hz,3H).

[0475] Compound 72: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-D-Val-Ala-Ala-Gln (acetate)

[0476] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4976([M+4H]) 4+ ), 549.6609([M+3H] 3+ ), 823.9876([M+2H] 2+ ).

[0477] 1 H NMR(600MHz,D2Oδ4.45–4.35(m,2H),4.26(dd,J=8.4,5.9Hz,1H),4.21–4.05(m,9 H),3.94–3.88(m,2H),3.82(t,J=6.8Hz,1H),3.73–3.57(m,3H),3.55–3.37(m,3H ),2.99(t,J=6.8Hz,2H),2.82–2.75(m,6H),2.23–2.06(m,9H),1.96–1.36(m,AcO H,50H),1.32–1.14(m,15H),0.78(dd,J=13.9,6.7Hz,6H),0.73(d,J=6.7Hz,6H).

[0478] Compound 73: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-D-Ala-Ala-Gln (acetate)

[0479] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4977([M+4H]) 4+ ), 549.6609([M+3H] 3+ ), 823.9879([M+2H] 2+ ).

[0480] 1H NMR(600MHz,D2O)δ4.48(dd,J=9.7,4.3Hz,1H),4.43(dd,J=9.3,4.9Hz,1H),4.32(dd,J=8.4,5.8Hz ,1H),4.27–4.23(m,3H),4.21–4.17(m,3H),4.15–4.10(m,3H),3.97(dd,J=9.2,4.6Hz,1H),3.89–3. 85(m,2H),3.76–3.66(m,3H),3.59–3.45(m,3H),3.05(t,J=6.9Hz,2H),2.87–2.81(m,6H),2.26(t, J=7.7Hz,2H),2.22–2.12(m,7H),2.00–1.48(m,AcOH,47H),1.37–1.20(m,16H),0.86–0.77(m,12H).

[0481] Compound 74: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-D-Ala-Gln (acetate)

[0482] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4977([M+4H]) 4+ ), 549.6610([M+3H] 3+ ), 823.9879([M+2H] 2+ ).

[0483] 1H NMR(600MHz,D2O)δ4.48(dd,J=9.6,4.3Hz,1H),4.44(dd,J=9.3,4.9Hz,1H),4.32(dd,J=8.4,5.8Hz,1H), 4.28–4.17(m,5H),4.17–4.09(m,4H),4.05(dd,J=9.1,4.7Hz,1H),3.95(d,J=7.7Hz,1H),3.88(t,J=6.8Hz ,1H),3.78–3.63(m,3H),3.60–3.37(m,3H),3.06(t,J=6.9Hz,2H),2.89–2.81(m,6H),2.29–2.09(m,9H), 2.02–1.46(m,AcOH,45H), 1.39–1.19(m,15H), 0.84(dd,J=14.0,6.7Hz,6H), 0.78(dd,J=14.3,6.8Hz,6H).

[0484] Compound 75: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-D-Gln (acetate)

[0485] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 127 N 23 O 20 m / z: 412.4978 ([M+4H]) 4+ ), 549.6611([M+3H] 3+ )823.9881([M+2H] 2+ ).

[0486] 1H NMR(600MHz,D2O)δ4.49–4.42(m,2H),4.32(dd,J=8.4,5.8Hz,1H),4.27–4.10(m,9H),4.04(dd, J=8.4,4.6Hz,1H),3.96(d,J=7.4Hz,1H),3.90–3.84(m,1H),3.73(d,J=7.7Hz,1H),3.71–3.64( m,2H),3.60–3.32(m,3H),3.06(t,J=6.9Hz,2H),2.89–2.79(m,6H),2.28–2.06(m,9H),1.99–1. 47(m,AcOH,44H),1.39–1.18(m,15H),0.84(dd,J=13.9,6.7Hz,6H),0.78(dd,J=6.8,3.3Hz,6H).

[0487] Compound 76: Glu-Gly-Val-Pro-Gln-Ala-Lys (acetate)

[0488] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 53 N9O 11 m / z: 364.7022([M+2H]) 2+ ), 728.3929([M+H] + ).

[0489] 1 H NMR(600MHz,D2O)δ4.30(d,J=7.8Hz,1H),4.26(dd,J=8.2,6.7Hz,1H),4.19–4.11(m,2H),4.02(dd, J=8.3,5.0Hz,1H),3.95(t,J=6.5Hz,1H),3.91–3.82(m,2H),3.78–3.72(m,1H),3.58–3.51(m,1H), 2.84(t,J=7.5Hz,2H),2.29–2.24(m,4H),2.20–2.11(m,1H),2.01–1.82(m,AcOH,10H),1.77–1.65( m,2H),1.60–1.50(m,3H),1.25(dd,J=7.5,4.7Hz,5H),0.84(d,J=6.8Hz,3H),0.80(d,J=6.7Hz,3H).

[0490] Compound 77: Glu-Pro-Val-Gly-Gln-Ala-Lys (acetate)

[0491] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 53 N9O 11 m / z: 364.7004([M+2H]) 2+ ), 728.3932([M+H] + ).

[0492] 1 H NMR(600MHz,D2O)δ4.40(dd,J=8.3,6.5Hz,1H),4.23(dd,J=7.7,4.4Hz,1H),4.20–4.15(m,2H ),4.01(dd,J=8.3,5.1Hz,1H),3.95(d,J=7.4Hz,1H),3.83–3.76(m,2H),3.63–3.55(m,1H),3 .50–3.45(m,1H),2.85(t,J=7.5Hz,2H),2.32(t,J=7.2Hz,2H),2.23–2.15(m,3H),2.05–1.67 (m,AcOH,12H),1.59–1.50(m,3H),1.26(dd,J=9.7,7.5Hz,5H),0.84(dd,J=14.3,6.8Hz,6H).

[0493] Compound 78: D-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0494] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N9O 11 m / z: 384.716([M+2H]) 2+ ), 768.4245([M+H] + ).

[0495] 1H NMR(600MHz,D2O)δ4.36(dd,J=8.8,4.3Hz,1H),4.29–4.21(m,3H),4.18–4.11( m,2H),4.01(dd,J=8.2,5.1Hz,1H),3.78–3.72(m,1H),3.63–3.49(m,3H),2.84( t,J=7.5Hz,2H),2.28–2.23(m,4H),2.19–2.12(m,2H),1.98–1.66(m,AcOH,15H) ,1.59–1.50(m,3H),1.25(dd,J=11.6,7.4Hz,5H),0.85(dd,J=15.5,6.8Hz,6H).

[0496] Compound 79: Glu-D-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0497] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N9O 11 m / z: 384.7159([M+2H]) 2+ ), 768.4244([M+H] + ).

[0498] 1 H NMR(600MHz,D2O)δ4.36(dd,J=8.8,3.9Hz,1H),4.33(d,J=8.1Hz,1H),4.28(dd,J=8.3, 6.6Hz,1H),4.23(dd,J=7.1,5.1Hz,1H),4.19–4.13(m,2H),4.02(dd,J=8.2,5.1Hz,1H) ,3.76–3.68(m,1H),3.61–3.51(m,3H),2.84(t,J=7.5Hz,2H),2.31–2.06(m,7H),2.00– 1.66(m,AcOH,16H),1.58–1.50(m,3H),1.27–1.23(m,5H),0.81(dd,J=22.8,6.7Hz,6H).

[0499] Compound 80: Glu-Pro-D-Val-Pro-Gln-Ala-Lys (acetate)

[0500] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N9O 11m / z: 384.7158([M+2H]) 2+ ), 768.4241([M+H] + ).

[0501] 1 H NMR(600MHz,D2O)δ4.46(d,J=7.3Hz,1H),4.36(dd,J=8.3,6.6Hz,1H),4.30–4.20(m,3H),4. 17(dd,J=8.1,6.1Hz,1H),4.03–4.00(m,1H),3.72–3.66(m,1H),3.63–3.57(m,2H),3.53–3. 49(m,1H),2.85(t,J=7.5Hz,2H),2.36–2.21(m,6H),2.18–2.13(m,1H),2.05–1.78(m,AcOH, 17H),1.70–1.66(m,1H),1.58–1.51(m,3H),1.28–1.23(m,6H),0.78(dd,J=15.0,6.7Hz,6H).

[0502] Compound 81: Glu-Pro-Val-D-Pro-Gln-Ala-Lys (acetate)

[0503] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 34 H 57 N9O 11 m / z: 384.7157([M+2H]) 2+ ), 768.4240([M+H] + ).

[0504] 1 H NMR(600MHz,D2O)δ4.43–4.30(m,2H),4.28–4.21(m,2H),4.21–4.13(m,2H),4 .03–3.98(m,1H),3.72–3.53(m,3H),3.50–3.44(m,1H),2.85(t,J=7.5Hz,2H) ,2.33(t,J=7.2Hz,2H),2.23–2.11(m,4H),2.07–1.74(m,AcOH,14H),1.73–1. 64(m,2H),1.58–1.50(m,3H),1.29–1.23(m,5H),0.81(dd,J=15.1,6.7Hz,6H).

[0505] Compound 82: Glu-Pro-Val-Pro-D-Gln-Ala-Lys (acetate)

[0506] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 53 N9O 11 m / z: 384.7158([M+2H]) 2+ ), 768.4242([M+H] + ).

[0507] 1 H NMR(600MHz,D2O)δ4.38(dd,J=8.3,6.4Hz,1H),4.27–4.14(m,5H),4.03(dd,J=8.4,5.0Hz,1H),3.80–3.75(m,1H),3.62–3.45(m,3H),2.85(t, J=7.5Hz,2H),2.32(t,J=7.3Hz,2H),2.24–2.12(m,4H),2.09–1.67(m,AcOH,15H),1.60–1.50(m,3H),1.28(d,J=7.3Hz,5H),0.88–0.80(m,6H).

[0508] Compound 83: Glu-Pro-Val-Pro-Gln-D-Ala-Lys (acetate)

[0509] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 53 N9O 11 m / z: 384.7159([M+2H]) 2+ ), 768.4243([M+H] + ).

[0510] 1H NMR(600MHz,D2O)δ4.38(dd,J=8.3,6.4Hz,1H),4.28–4.20(m,3H),4.19–4.12(m,2H),4 .05(dd,J=8.6,5.0Hz,1H),3.77–3.72(m,1H),3.60–3.45(m,3H),2.85–2.80(m,2H),2. 32(t,J=7.2Hz,2H),2.26(t,J=8.0Hz,2H),2.20–2.12(m,2H),2.08–1.77(m,AcOH,13H) ,1.75–1.66(m,3H),1.57–1.47(m,3H),1.26–1.18(m,5H),0.85(dd,J=13.0,6.7Hz,6H).

[0511] Compound 84: Glu-Pro-Val-Pro-Gln-Ala-D-Lys (acetate)

[0512] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 53 N9O 11 m / z: 384.7160([M+2H]) 2+ ), 768.4246([M+H] + ).

[0513] 1 H NMR(600MHz,D2O)δ4.39(dd,J=8.3,6.4Hz,1H),4.30–4.20(m,3H),4.19–4.11(m,2H),4 .05(dd,J=8.3,4.8Hz,1H),3.79–3.71(m,1H),3.61–3.47(m,3H),2.86–2.81(m,2H),2. 32(t,J=7.2Hz,2H),2.27(t,J=7.5Hz,2H),2.21–2.12(m,2H),2.05–1.78(m,AcOH,12H) ,1.78–1.68(m,3H),1.60–1.50(m,3H),1.27–1.20(m,5H),0.86(dd,J=12.5,6.7Hz,6H).

[0514] Compound 85: Lys-Glu-Pro-Val-Pro (acetate)

[0515] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 26 H 44N6O8; m / z: 454.2655 ([M+2H]) 2+ ), 569.3287([M+H] + ).

[0516] 1 H NMR (600MHz, D2O) δ4.50 (dd, J=9.9, 4.2Hz, 1H), 4.36–4.22 (m, 3H), 4.09 (dd, J=8. 5,5.8Hz,1H),3.89(t,J=6.7Hz,1H),3.70–3.65(m,2H),3.60–3.53(m,2H),3.51–3 .26(m,1H),2.87(d,J=7.5Hz,2H),2.28–2.09(m,5H),2.00–1.68(m,AcOH,18H),1 .58–1.55(m,2H),1.35–1.29(m,2H),0.88(d,J=6.8Hz,3H),0.82(d,J=6.7Hz,3H).

[0517] Compound 86: Glu-Pro-Val-Pro-Gln (acetate)

[0518] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 25 H 40 N6O9; m / z: 569.2919 ([M+H]) + ).

[0519] 1 H NMR(600MHz,D2O)δ4.40(dd,J=8.3,6.4Hz,1H),4.31–4.23(m,3H),4.09(dd,J=9.0,4.8Hz,1H),3.78–3.72(m,1H),3 .61–3.48(m,3H),2.41(t,J=7.3Hz,2H),2.25–2.14(m,4H),2.09–1.70(m,AcOH,13H),0.86(dd,J=15.3,6.7Hz,6H).

[0520] Compound 87: Lys-Glu-Pro-Val-Pro-Gln (acetate)

[0521] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 31 H 52 N8O 10 m / z: 349.1971([M+2H]) 2+ ), 697.3868([M+H]+ ).

[0522] 1 H NMR(600MHz,D2O)δ4.51(dd,J=9.8,4.3Hz,1H),4.34–4.25(m,3H),4.03(dd, J=8.8,4.7Hz,1H),3.89(t,J=6.7Hz,1H),3.79–3.65(m,2H),3.63–3.52(m,2H ),2.86(t,J=7.5Hz,2H),2.28–2.13(m,6H),2.00–1.73(m,AcOH,17H),1.60– 1.54(m,2H),1.36–1.28(m,2H),0.86(d,J=6.8Hz,3H),0.83(d,J=6.6Hz,3H).

[0523] Compound 88: Val-Pro-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0524] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 92 H 152 N 26 O 24 m / z: 502.2934 ([M+4H]) 4+ ), 669.3699([M+3H] 3+ ).

[0525] 1H NMR(600MHz,D2O)δ6.97–6.94(m,2H),6.70–6.66(m,2H),4.44(dd,J=9.4,4.8Hz,1H),4.35–4.08(m,17H),4.05–4.00 (m,2H),3.96(d,J=7.6Hz,1H),3.76–3.67(m,3H),3.64–3.53(m,3H),3.48(dd,J=10.2,6.3Hz,3H),3.07(t,J=6.9Hz,2 H),2.94(dd,J=13.6,6.6Hz,1H),2.88–2.79(m,7H),2.76(dd,J=13.6,9.2Hz,1H),2.27(t,J=7.6Hz,2H),2.21(t,J=7. 6Hz,2H),2.18–2.08(m,8H),2.01–1.45(m,AcOH,59H),1.30–1.10(m,16H),0.95(d,J=6.9Hz,3H),0.87–0.77(m,18H).

[0526] Compound 89: pyro-Glu-Leu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0527] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 84 H 145 N 25 O 24 m / z: 468.7766([M+4H]) 4+ ), 624.6998([M+3H] 3+ ), 936.5460([M+2H] 2+ ).

[0528] 1H NMR(600MHz,D2O)δ4.47–4.42(m,2H),4.32–4.10(m,13H),4.02(dd,J=8.4,4.8Hz,1H),3.9 6(d,J=7.6Hz,1H),3.82–3.60(m,3H),3.58–3.45(m,3H),3.07(t,J=6.9Hz,2H),2.90–2.80( m,6H),2.45–2.36(m,1H),2.31–2.24(m,4H),2.23–2.03(m,7H),1.99–1.44(m,AcOH,47H), 1.39–1.19(m,15H),0.85(dd,J=12.5,6.8Hz,6H),0.81–0.77(m,9H),0.75(d,J=6.0Hz,3H).

[0529] Compound 90: Pro-Ala-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0530] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 90 H 148 N 26 O 24 m / z: 495.2855([M+4H]) 4+ ), 660.0449([M+3H] 3+ ), 989.5637([M+2H] 2+ ).

[0531] 1 H NMR(600MHz,D2O)δ6.99–6.92(m,2H),6.71–6.64(m,2H),4.47–4.42(m,1H),4.36(t,J =7.8Hz,1H),4.32–4.06(m,15H),4.04–3.99(m,1H),3.96(d,J=7.6Hz,1H),3.79–3.65( m,3H),3.60–3.46(m,3H),3.30–3.22(m,2H),3.07(t,J=6.9Hz,2H),2.89–2.79(m,8H), 2.33–2.06(m,11H),1.99–1.45(m,AcOH,51H),1.39–1.14(m,19H),0.87–0.77(m,12H).

[0532] Compound 91: Arg-Lys-Asp-Val-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0533] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 103 H 174 N 32 O 28 m / z: 578.0856 ([M+4H]) 4+ ), 770.4463([M+3H] 3+ ), 1155.1642([M+2H] 2+ ).

[0534] 1 H NMR(600MHz,D2O)δ6.96(d,J=8.5Hz,2H),6.70–6.64(m,2H),4.50–4.43(m,2H),4.38(t,J=8.0Hz,1H),4.30–4.11( m,13H),4.02(dd,J=8.4,4.9Hz,1H),3.96(d,J=7.6Hz,1H),3.94–3.90(m,2H),3.79–3.65(m,3H),3.62–3.46(m,3H ),3.11–3.05(m,4H),2.88–2.79(m,10H),2.51(dd,J=15.9,5.3Hz,1H),2.42(dd,J=16.0,8.5Hz,1H),2.29–2.11(m ,9H),1.99–1.64(m,AcOH,45H),1.63–1.45(m,19H),1.41–1.14(m,18H),0.87–0.77(m,12H),0.72(d,J=6.7Hz,6H).

[0535] Compound 92: Arg-Lys-Asp-Val-Tyr-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0536] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 70 H 116 N 20 O 20 m / z: 390.2233([M+4H]) 4+ ), 519.9619([M+3H] 3+ ), 779.4393([M+2H]2+ ).

[0537] 1 H NMR(600MHz,D2O)δ6.96(d,J=8.6Hz,2H),6.67(d,J=8.5Hz,2H),4.49(dd,J=8.4,5.3Hz,1H),4.38(t,J=8.0Hz,1H),4.31–4.23 (m,5H),4.19–4.10(m,3H),4.01(dd,J=8.2,5.1Hz,1H),3.93–3.89(m,2H),3.77–3.67(m,2H),3.61–3.50(m,2H),3.10–3.05(m ,2H),2.86–2.79(m,8H),2.51(dd,J=16.0,5.3Hz,1H),2.42(dd,J=15.9,8.5Hz,1H),2.28–2.20(m,4H),2.18–2.10(m,2H),1.9 7–1.65(m,AcOH,29H),1.61–1.45(m,13H),1.26(dd,J=11.1,7.4Hz,10H),0.83(dd,J=19.0,6.7Hz,6H),0.72(d,J=5.6Hz,6H).

[0538] Compound 93: Arg-Lys-Asp-Val-Tyr-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0539] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 64 H 104 N 18 O 19 m / z: 715.3917([M+2H]) 2+ ).

[0540] 1H NMR(600MHz,D2O)δ6.96(d,J=8.6Hz,2H),6.66(d,J=8.5Hz,2H),4.48–4.42(m,3H),4.26–4.12(m,6H),4.02(dd,J=8.2, 5.0Hz,1H),3.95–3.89(m,2H),3.77–3.68(m,1H),3.56–3.47(m,2H),3.43–3.36(m,1H),3.10–3.04(m,2H),2.87–2.80(m ,6H),2.54(dd,J=16.0,5.1Hz,1H),2.45(dd,J=16.0,8.4Hz,1H),2.27(t,J=7.6Hz,2H),2.17–2.08(m,4H),1.98–1.66(m ,AcOH,27H),1.65–1.45(m,9H),1.26(dd,J=11.3,7.4Hz,7H),0.84(dd,J=13.0,6.7Hz,6H),0.70(dd,J=6.8,2.8Hz,6H).

[0541] Compound 94: Arg-Lys-Asp-Val-Tyr-Pro-Val-Pro-Gln (acetate)

[0542] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 50 H 80 N 14 O 14 m / z: 367.8727([M+3H]) 3+ ), 551.3051([M+2H] 2+ ), 1101.6030([M+H] + ).

[0543] 1H NMR(600MHz,D2O)δ4.45(dd,J=8.9,4.8Hz,1H),4.36–4.22(m,4H),4.04–3.98(m,1H),3.90(dd,J=7.0,5.8H z,2H),3.76(dd,J=10.1,6.4Hz,1H),3.69–3.63(m,1H),3.60–3.43(m,2H),3.09–3.05(m,2H),2.95(dd,J=14 .3,5.2Hz,1H),2.86–2.82(m,2H),2.74–2.67(m,1H),2.52–2.31(m,2H),2.22–2.08(m,4H),1.99–1.46(m,A cOH,29H),1.32–1.23(m,2H),0.90–0.81(m,6H),0.75(dd,J=10.3,6.8Hz,2H),0.67(dd,J=17.1,6.7Hz,4H).

[0544] Compound 95: Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0545] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 113 H 188 N 34 O 31 m / z: 504.6906([M+5H]) 5+ ), 630.6115([M+4H] 4+ ), 840.4796([M+3H] 3+ ).

[0546] 1H NMR(600MHz,D2O)δ7.22(t,J=7.4Hz,2H),7.16(t,J=7.3Hz,1H),7.12–7.07(m,2H),4.47–4.41(m,3H),4.35–4. 03(m,19H),4.02(dd,J=8.4,4.8Hz,1H),3.95(d,J=3.6Hz,1H),3.86(s,1H),3.75(s,1H),3.68(t,J=8.5Hz,2H), 3.58–3.42(m,5H),3.06(t,J=6.9Hz,4H),2.91–2.81(m,7H),2.29–2.11(m,12H),2.00–1.19(m,AcOH,80H),1.14 (d,J=7.1Hz,3H),1.06(d,J=6.3Hz,3H),0.84(dd,J=13.7,6.7Hz,6H),0.81–0.76(m,9H),0.72(d,J=5.6Hz,3H).

[0547] Compound 96: Gly-Pro-Glu-Thr-Ala-Phe-Leu-Arg-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0548] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 74 H 118 N 20 O 22 m / z: 547.2972 ([M+3H]) 3+ ), 820.4424([M+2H] 2+ ).

[0549] 1H NMR(600MHz,D2O)δ7.21(dd,J=8.2,6.6Hz,2H),7.16(t,J=7.3Hz,1H),7.12–7.07(m,2H),4.50–4.42(m,2H),4.36–4.22(m,4H),4.21–4 .09(m,7H),4.07–4.00(m,2H),3.93(d,J=16.5Hz,1H),3.84(d,J=16.5Hz,1H),3.78–3.70(m,1H),3.68–3.60(m,1H),3.56–3.42(m,4H), 3.10–2.97(m,3H),2.90–2.82(m,3H),2.29–2.09(m,9H),2.00–1.81(m,AcOH,19H),1.77–1.66(m,5H),1.61–1.37(m,9H),1.26(dd,J=11 .9,7.4Hz,5H),1.14(d,J=7.2Hz,3H),1.05(d,J=6.4Hz,3H),0.84(dd,J=15.5,6.7Hz,6H),0.77(d,J=5.9Hz,3H),0.72(d,J=5.7Hz,3H).

[0550] Compound 97: Ser-Ser-Glu-Asp-Ile-Lys-Glu-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0551] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 105 H 179 N 31 O 35 m / z: 488.0708([M+5H]) 5+ ), 609.8367([M+4H] 4+ ), 812.7799([M+3H] 3+ ).

[0552] 1H NMR(600MHz,D2O)δ4.48–4.41(m,3H),4.34(t,J=5.4Hz,1H),4.31(dd,J=8.4,5.8Hz,1H),4.28–4.22(m ,3H),4.22–4.09(m,11H),4.04–3.88(m,5H),3.81–3.72(m,3H),3.71–3.62(m,2H),3.59–3.43(m,3H),3 .07(t,J=6.9Hz,2H),2.90–2.80(m,8H),2.58(dd,J=16.0,6.3Hz,1H),2.48(dd,J=16.0,7.9Hz,1H),2. 29–2.09(m,13H),2.00–1.47(m,AcOH,51H),1.40–1.20(m,18H),1.12–1.05(m,1H),0.89–0.70(m,18H).

[0553] Compound 98: Ser-Ser-Glu-Asp-Ile-Lys-Glu-Glu-Pro-Val-Pro-Gln-Ala-Lys (acetate)

[0554] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 66 H 109 N 17 O 26 m / z: 519.5973 ([M+3H]) 3+ ), 778.8925([M+2H] 2+ ).

[0555] 1 H NMR(600MHz,D2O)δ4.54–4.47(m,2H),4.37–4.30(m,2H),4.27–4.10(m,8H),4.05–3.99(m,2H),3 .94–3.88(m,2H),3.79–3.73(m,3H),3.70–3.62(m,1H),3.61–3.50(m,2H),2.85(t,J=7.6Hz,4H) ,2.63(dd,J=16.2,6.2Hz,1H),2.51(dd,J=16.2,7.9Hz,1H),2.37–2.06(m,11H),1.99–1.49(m,A cOH,30H),1.34–1.24(m,8H),1.10–1.01(m,1H),0.84(dd,J=13.8,6.7Hz,6H),0.77–0.71(m,6H).

[0556] Compound 99: Ser-Ser-Glu-Asp-Ile-Lys-Glu-Pro-Val-Pro-Gln (acetate)

[0557] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 52 H 85 N 13 O 21 m / z: 614.806([M+2H]) 2+ ), 1228.605([M+H] + ).

[0558] 1 H NMR(600MHz,D2O)δ4.54–4.48(m,2H),4.36(t,J=5.5Hz,1H),4.34–4.24(m,3H),4.23–4.18(m,2H),4.12– 4.05(m,2H),4.01(d,J=7.9Hz,1H),3.94–3.88(m,2H),3.79–3.72(m,3H),3.70–3.62(m,1H),3.60–3.50( m,2H),2.85(t,J=7.6Hz,2H),2.65(dd,J=16.3,6.0Hz,1H),2.53(dd,J=16.4,8.0Hz,1H),2.40–2.03(m,9 H),2.01–1.51(m,AcOH,21H),1.34–1.22(m,3H),1.08–1.01(m,1H),0.89–0.80(m,6H),0.76–0.70(m,6H).

[0559] Compound 100: Val-Pro-Pro-Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0560] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 88 H 150 N 26 O 23 m / z: 389.0348 ([M+5H]) 5+ ), 480.0415([M+4H] 4+ ), 647.7197([M+3H] 3+ ).

[0561] 1H NMR(600MHz,D2O)δ4.49–4.42(m,2H),4.34–4.06(m,13H),4.05–4.00(m,2H),3.96(d, J=7.6Hz,1H),3.80–3.73(m,1H),3.69(d,J=9.8Hz,4H),3.58–3.45(m,5H),3.07(t,J= 6.9Hz,2H),2.88–2.83(m,6H),2.27(t,J=7.7Hz,3H),2.20–2.11(m,9H),2.01–1.45(m ,AcOH,55H),1.24(dd,J=21.5,7.2Hz,16H),0.97(d,J=6.9Hz,3H),0.88–0.78(m,15H).

[0562] Compound 101: Glu-Pro-Val-Pro-Gln-Ala-Lys-Ser-Ser-Glu-Asp-Ile-Lys-Glu (acetate)

[0563] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 66 H 109 N 17 O 26 m / z: 519.5978 ([M+3H]) 3+ ), 778.8933([M+2H] 2+ ).

[0564] 1 H NMR(600MHz,D2O)δ4.50(dd,J=8.0,6.0Hz,1H),4.40(dd,J=8.4,6.4Hz,1H),4.36–4.30(m,2H),4.27–4.19(m,6H), 4.18–4.11(m,2H),4.08(dd,J=8.8,4.9Hz,1H),4.01(d,J=8.1Hz,1H),3.82–3.71(m,5H),3.62–3.47(m,3H),2.86(t ,J=7.5Hz,4H),2.65(dd,J=16.4,6.0Hz,1H),2.51(dd,J=16.3,8.1Hz,1H),2.34(t,J=7.2Hz,2H),2.31–2.10(m,9H ),2.06–1.52(m,AcOH,28H),1.35–1.23(m,8H),1.08–1.00(m,1H),0.86(dd,J=11.5,6.7Hz,6H),0.76–0.70(m,6H).

[0565] Compound 102Glu-Pro-Val-Pro-Gln-Ala-Lys-Val-Pro-Tyr (acetate)

[0566] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 53 H 82 N 12 O 15 m / z: 564.3073([M+2H]) 2+ ).

[0567] 1 H NMR(600MHz,D2O)δ7.01–6.95(m,2H),6.69(d,J=8.5Hz,2H),4.40(dd,J=8.3,6.4Hz ,1H),4.29–4.10(m,9H),3.79–3.71(m,1H),3.68–3.45(m,5H),2.92–2.81(m,4H),2 .33(t,J=7.3Hz,2H),2.26(t,J=7.7Hz,2H),2.20–2.13(m,2H),2.06–1.52(m,AcOH, 25H),1.33–1.21(m,5H),0.86(dd,J=11.3,6.7Hz,6H),0.77(dd,J=17.6,6.7Hz,6H).

[0568] Compound 103: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Glu (acetate)

[0569] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 126 N 22 O 21 m / z: 824.4799([M+2H]) 2+ ).

[0570] 1H NMR (600MHz, D2O) δ4.49 (dd, J=9.6, 4.4Hz, 1H), 4.45 (dd, J=9.4, 4.9Hz, 1H), 4.36–4. 05(m,11H),4.01–3.96(m,2H),3.92–3.87(m,1H),3.82–3.64(m,3H),3.60–3.46(m,3 H),3.07(t,J=7.0Hz,2H),2.88–2.83(m,6H),2.29–2.09(m,9H),1.99–1.46(m,AcOH, 47H),1.39–1.20(m,16H),0.85(dd,J=14.2,6.7Hz,6H),0.80(dd,J=6.7,3.7Hz,6H).

[0571] Compound 104: Lys-Glu-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Asn (acetate)

[0572] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 72 H 125 N 23 O 20 m / z: 408.9938([M+4H]) 4+ ), 544.9891([M+3H] 3+ ), 816.9801([M+2H] 2+ ).

[0573] 1 H NMR(600MHz,D2O)δ4.49(dd,J=9.7,4.4Hz,1H),4.45(dd,J=9.4,4.8Hz,1H),4.34–4.10(m,11H),3.9 6(d,J=7.6Hz,1H),3.92–3.84(m,1H),3.78–3.66(m,3H),3.62–3.43(m,3H),3.07(t,J=6.9Hz,2H),2 .90–2.82(m,6H),2.63(dd,J=15.1,4.9Hz,1H),2.53(dd,J=15.1,8.1Hz,1H),2.30–2.11(m,7H),2.0 3–1.42(m,AcOH,44H),1.39–1.19(m,15H),0.85(dd,J=14.0,6.7Hz,6H),0.80(dd,J=6.8,3.0Hz,6H).

[0574] Compound 105: Lys-Gln-Pro-Val-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0575] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 73 H 128 N 24 O 19 m / z: 412.2515([M+4H]) 4+ ), 549.3327([M+3H] 3+ ), 823.4955([M+2H] 2+ ).

[0576] 1 H NMR(600MHz,D2O)δ4.53(dd,J=9.0,5.0Hz,1H),4.44(dd,J=9.4,4.8Hz,1H),4.34(dd,J=8.4,6.0Hz,1H),4.30 –4.23(m,3H),4.22–4.11(m,6H),4.02(dd,J=8.4,4.8Hz,1H),3.96(d,J=7.6Hz,1H),3.90(t,J=6.7Hz,1H),3. 80–3.63(m,3H),3.60–3.42(m,3H),3.07(t,J=6.9Hz,2H),2.89–2.82(m,6H),2.34–2.24(m,4H),2.22–2.07(m ,5H),2.01–1.45(m,AcOH,50H),1.39–1.20(m,15H),0.85(dd,J=15.5,6.7Hz,6H),0.80(dd,J=6.8,4.3Hz,6H).

[0577] Compound 106: Lys-Glu-Pro-Ile-Pro-Gln-Ala-Lys-Pro-Arg-Lys-Val-Ala-Ala-Gln (acetate)

[0578] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 74 H 129 N 23 O 20 m / z: 416.0015([M+4H]) 4+ ), 554.3328([M+3H] 3+ ), 830.9956([M+2H] 2+ ).

[0579] 1 H NMR(600MHz,D2O)δ4.50(dd,J=9.6,4.4Hz,1H),4.45(dd,J=9.3,4.9Hz,1H),4.36–4.22(m,4H),4.22–4.1 0(m,6H),4.02(dd,J=8.4,4.9Hz,1H),3.96(d,J=7.6Hz,1H),3.92–3.84(m,1H),3.82–3.74(m,1H),3.73–3 .63(m,2H),3.61–3.41(m,3H),3.07(t,J=6.9Hz,2H),2.90–2.82(m,6H),2.32–2.12(m,9H),2.00–1.46(m, AcOH,46H),1.43–1.05(m,17H),0.84(d,J=6.8Hz,3H),0.80(dd,J=6.8,4.3Hz,6H),0.75(t,J=7.4Hz,3H).

[0580] Compound 107: Val-Pro-Gln-Ala (acetate)

[0581] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 18 H 31 N5O6; m / z: 414.2341 ([M+1H]) + ).

[0582] 1 H NMR(600MHz,D2O)δ4.29(t,J=7.6Hz,1H),4.07(dd,J=8.5,6.1Hz,1H),4.01–3.91(m,2H),3.61–3.53(m,1H),3.46–3.41 (m,1H),2.24–2.10(m,4H),1.95–1.69(m,AcOH,6H),1.14(d,J=7.3Hz,3H),0.91(d,J=7.0Hz,3H),0.80(d,J=6.8Hz,3H).

[0583] Compound 108: Val-Pro-Gln-Ala-Lys (acetate)

[0584] High-resolution mass spectrometry (Orbitrap-HRMS), molecular formula: C 24 H 43 N7O7; m / z: 271.6681([M+2H]2+), 542.3289([M+1H] + ).

[0585] 1 1H NMR (600 MHz, D2O) δ 4.28 (t, J = 7.6 Hz, 1H), 4.13–4.06 (m, 2H), 3.99 (d, J = 5.5 Hz, 1H), 3.94 (dd, J = 8.2, 5.1 Hz, 1H), 3.60–3.54 (m, 1H), 3.46–3.41 (m, 1H), 2.78 (t, J = 7.5 Hz, 2H), 2.21 (t, J = 7.6 Hz, 2H), 2.17–2.09 (m, 2H), 1.93–1.58 (m, AcOH, 11H), 1.53–1.44 (m, 3H), 1.20 (dd, J = 9.7, 7.5 Hz, 5H), 0.91 (d, J = 6.9 Hz, 3H), 0.80 (d, J = 6.9 Hz, 3H).

[0586] Example 2: Experiment on the efficacy of polypeptide compounds in alleviating colitis

[0587] Experimental animals: Wild-type AB strain zebrafish, reproduced by natural paired mating. Zebrafish at 3 days post-fertilization (3 dpf). All zebrafish were raised in fish-raising water at 28 °C (water quality: 200 mg of instant sea salt was added to every 1 L of reverse osmosis water, with a conductivity of 450 - 550 μS / cm; pH of 6.5 - 8.5; hardness of 50 - 100 mg / L CaCO3). The license number for the use of experimental animals is: SYXK(Zhe)2012 - 0171, and the feeding management meets the requirements of international AAALAC certification (certification number: 001458).

[0588] Experimental methods:

[0589] (1) Grouping: Polypeptide group: Prepared into a 20.0 mg / mL stock solution with ultrapure water and stored at -20 °C.

[0590] Positive control group: Prednisone, white powder, batch number C10016501, Shanghai Macklin Biochemical Co., Ltd., stored at 4 °C; prepared into a 15.0 mg / mL stock solution with DMSO and stored in aliquots at -20 °C.

[0591] Normal control group and model control group.

[0592] (2) Experimental Methods: 3dpf wild-type AB strain zebrafish were randomly selected and placed in 6-well plates, with 30 zebrafish treated in each well (experimental group). Except for the normal control group, all experimental groups were treated with TNBS dissolved in water to establish a zebrafish intestinal mucosal injury model. After treatment at 28℃ for 2 days, the TNBS was removed, and polypeptide compounds (concentrations shown in Table 4) were dissolved in water and administered. Prednisone 15.0 μg / mL was used as a positive control. A normal control group (healthy group) and a model control group (intestinal mucosal injury group) were also set up, with a volume of 3 mL per well (experimental group). After further treatment at 28℃ for 2 days, 10 zebrafish from each experimental group were randomly selected and photographed under a dissecting microscope. Data were analyzed and collected using NIS-Elements D 3.20 advanced image processing software. The intestinal lumen area of ​​the zebrafish was analyzed, and the statistical analysis results of this index were used to evaluate the efficacy of the samples in alleviating colitis. Statistical results are expressed as mean ± SE. Statistical analysis was performed using SPSS 26.0 software; p < 0.05 indicated statistical significance.

[0593] Experimental results: The efficacy of polypeptide compounds in relieving colitis is shown in Table 4.

[0594] Table 4. Experimental results evaluating the efficacy of polypeptide compounds in relieving colitis (n=10)

[0595]

[0596] Compared with the model control group, *p<0.05, **p<0.01, ***p<0.001

[0597] As shown in Table 4, the polypeptide compound groups (compounds 1, 4, 5, 6, 12, and 15) exhibited significant statistical differences compared to the model control group (p<0.05; p<0.01; p<0.001), and their effects were superior to the prednisone group. The experimental results indicate that the polypeptide compounds provided in this application have the effect of relieving colitis, specifically by significantly alleviating intestinal dilatation.

[0598] The experimental system was designed with test substance concentration gradients of 250 μg / mL, 500 μg / mL, and 1000 μg / mL. By statistically analyzing the intestinal lumen area of ​​zebrafish after intervention in each group, it was found that most test substances with colitis-relieving effects showed significant effects at a concentration of 1000 μg / mL (p<0.05; p<0.01; p<0.001). Further increasing the test substance concentration resulted in zebrafish mortality within the groups. Therefore, in subsequent rounds of activity evaluation, all test substance peptide compounds were uniformly administered at 1000 μg / mL. The results are summarized in Table 5.

[0599] Table 5 Summary of the activity of peptide compounds in alleviating intestinal dilation in zebrafish colitis.

[0600]

[0601] Note: The activity of each compound in alleviating intestinal distension in zebrafish colitis was determined through multiple batches of experiments. For ease of comparison, its anti-intestinal distension activity was compared with the control group of the same batch: *p<0.05, **p<0.01, ***p<0.001; ND indicates not tested, "—" indicates no activity, "+" indicates some effect in alleviating intestinal distension, "#" indicates that all zebrafish died in the experiment, and "&" indicates that 13 zebrafish died in the experiment.

[0602] According to the data in Tables 4 and 5, the peptide group showed significant statistical differences compared to the model control group (p<0.05, p<0.01, or p<0.001), and the effect was better than that of the prednisone group. Under the experimental conditions, compounds 1, 4-6, 9, 10, 12, 15, 25, 26, 29, 31, 34, 38, 48-51, 53-55, 57, 62, 63, 65-68, 70, 72, 75, 83, 85, 86, 88-90, 92-100, and 103-106 all showed significant efficacy in relieving colitis, specifically by alleviating intestinal distension (p<0.05); compounds 7, 8, 11, 14, 16, 17, 19, 21-24, 27, 28, 30, 32, 33, 36, 37, 39-42, 46, 47, 52, 58, 59, 64, 69, 71, 74, 76, 78-82, 84, 87, 91, and 102 had a certain effect in relieving intestinal distension.

[0603] Example 3: Evaluation of the therapeutic efficacy of peptide compounds on a TNBS-induced acute inflammatory bowel disease (UC) model in SD rats.

[0604] Objective: To evaluate the therapeutic effect of peptide compounds on a TNBS-induced acute inflammatory bowel disease model in SD rats.

[0605] Materials and Methods:

[0606] This experiment established a rat model of ulcerative colitis by administering a single enema of trinitrobenzenesulfonic acid (TNBS) to male SD rats. TNBS-induced acute colitis in rats is an immune response induced by cytokines secreted by Th1 cells. In the control group, the colon showed significant atrophy and intestinal wall thickening, with a significantly increased colon weight-to-length ratio. Cleft-like ulcer foci were visible on the intestinal mucosal surface, and typical granulomas were observed in the submucosa and muscularis propagules, accompanied by infiltration of inflammatory cells, primarily lymphocytes and plasma cells. Simultaneously, significant proliferation of small blood vessels and connective tissue (fibroblasts) was observed, and crypt structure destruction and hemorrhage were visible around the ulcer foci. This model is simple to establish, has good reproducibility, and is widely used as an experimental animal model for evaluating drugs for treating inflammatory bowel disease. Experimental animals (male SD rats) were randomly divided into six groups based on body weight on the first day of the experiment: a normal control group, a model control group, a positive control group treated with mesalazine, a low-dose treatment group for compound 1, a high-dose treatment group for compound 1, a low-dose treatment group for compound 6, a high-dose treatment group for compound 6, and a treatment group for compound 12, with 10 rats per group. On the day of grouping (D0), except for the normal control group which received a solvent, all other experimental groups were induced to develop the model by enema with TNBS ethanol solution. Treatment with the test drugs and positive control drugs began the day after model induction. The treatment doses of compound 1 were 3.0 mg / kg and 6.0 mg / kg, the treatment doses of compound 6 were 1.4 mg / kg and 2.8 mg / kg, the treatment dose of compound 12 was 0.8 mg / kg, and the treatment dose of the positive control drug mesalazine was 30.0 mg / kg. All drugs were administered via enema once daily for 14 consecutive days. The experimental treatment period was (…). During days 1-14, dynamic data were collected on the body weight and general clinical observation indicators (including but not limited to food intake, water intake, activity level, and defecation) of animals in each experimental group. Simultaneously, based on the ulcerative colitis disease activity-related scoring criteria, the degree of weight loss, fecal morphology, and fecal hemorrhage response after TNBS induction were scored for each experimental group, and the ulcerative colitis disease activity (DAI) was calculated. Data were collected daily for 14 consecutive days. At the experimental endpoint, colonic tissue was collected from each experimental group. After measuring colonic length and weighing, images were acquired without necropsy. Subsequently, the colon was longitudinally dissected along the intestinal wall, and necropsy images were acquired. Gross lesions such as colonic adhesions, colonic ulcers, and inflammation in each experimental group were observed and scored based on the "Coronary Gross Morphology Scoring Criteria." Ulcer images were acquired under a stereomicroscope, and the area of ​​colonic ulcer foci was calculated. Routine fixation was then performed. Based on the statistical analysis results of various pharmacodynamic indicators, the therapeutic effect of the tested peptide compound on the ulcerative colitis disease model under the established treatment regimen was evaluated.

[0607] Results and conclusions:

[0608] In this experimental study, the model control group animals exhibited obvious disease characteristics of ulcerative colitis, including inhibited weight gain, soft and loose stools, significantly elevated DAI scores for ulcerative colitis disease activity, and gross observation revealed colonic atrophy, typical ulcer foci, and confluent colonic ulcer foci. Compound 1, under the established treatment regimen, significantly improved the inhibited weight gain status in the model animals, with a statistically significant difference in the degree of improvement compared to the control group (P<0.05). Compounds 6 and 12, however, did not show significant improvement in the inhibited weight gain status of the model animals.

[0609] Table 6. Effects of the tested peptide compounds on body weight changes in model animals.

[0610]

[0611]

[0612] Note: All data are presented as mean ± standard deviation (SD) for statistical analysis. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 indicates the statistical analysis results of the model control group and the normal control group. # P<0.05, ## P<0.01 indicates the statistical analysis results of the animals in the treatment group and the model control group compared with those in the test polypeptide compound treatment group.

[0613] The DAI scores during the experimental period showed that the tested compounds 1, 6, and 12 had a significant ameliorative effect on the progression of ulcerative colitis in the model animals under the set treatment regimen. The fecal characteristics and weight loss of the animals in the corresponding treatment groups were significantly alleviated, and the mean DAI scores of the animals all showed a decrease. Compared with the model control group at the same time, there was a significant statistical difference (P<0.05). The therapeutic effect of compound 1 was the most significant. At the same time, the therapeutic effects of compounds 1 and 6 showed a clear dose-dependent effect.

[0614] Table 7. Effects of the tested peptide compounds on the DAI score of model animals.

[0615]

[0616] Note: All data are presented as mean ± standard deviation (SD) for statistical analysis. * P<0.05, ** P<0.01, *** P<0.001, ****P<0.0001 indicates the statistical analysis results of the model control group and the normal control group. # P<0.05, ## P<0.01, ### P<0.001, #### P<0.0001 indicates the statistical analysis results of the animals in the treatment group with the tested polypeptide compound and the model control group. + P<0.05, ++ P<0.01, +++ P<0.001, ++++ P<0.0001 indicates the statistical analysis results between the dosage groups of compound 1 or 6.

[0617] The results of the experimental endpoints, including gross anatomy and colonic tissue damage scores, showed that compounds 1, 6, and 12 had significant therapeutic effects on TNBS-induced gross colonic damage (colonic adhesions, ulcers, and inflammation), colonic ulcer area, and abnormal increase in colonic weight / length ratio. The degree of improvement was statistically different from that of the control group at the same time point (P<0.05). Compound 1 had the most significant therapeutic effect, and its therapeutic effect showed a clear dose-dependent effect.

[0618] Table 8. Effects of the tested polypeptide compounds on gross colonic injury and ulcer area in model animals.

[0619]

[0620]

[0621] Note: All data are presented as mean ± standard deviation (SD) for statistical analysis. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 indicates the statistical analysis results of the model control group and the normal control group. # P<0.05, ## P<0.01, ### P<0.001, #### P<0.0001 indicates the statistical analysis results of the animals in the treatment group with the tested polypeptide compound and the model control group. + P<0.05, ++ P<0.01, +++ P<0.001, ++++ P<0.0001 indicates the statistical analysis results between the dosage groups of compound 1 or 6.

[0622] Table 9. Effects of the tested polypeptide compounds on the colon weight / length ratio in model animals.

[0623]

[0624] Note: All data are presented as mean ± standard deviation (SD) for statistical analysis. * P<0.05, ** P<0.01, *** P<0.001, **** P<0.0001 indicates the statistical analysis results of the model control group and the normal control group. # P<0.05, ## P<0.01, ### P<0.001, #### P<0.0001 indicates the statistical analysis results of the animals in the treatment group with the tested polypeptide compound and the model control group. + P<0.05, ++ P<0.01, +++ P<0.001, ++++ P<0.0001 indicates the statistical analysis results between the dosage groups of compound 1 or 6.

[0625] This experiment used TNBS to induce colonic lesions in male SD rats via enema, establishing a rat model of ulcerative colitis consistent with clinical pathogenesis. Treatment with test compounds 1, 6, and 12, and the positive control drug mesalazine, began the day after model induction. During the treatment period (D1-D14), changes in animal body weight, DAI score (disease activity index), gross colonic damage (colonic adhesions, ulcers, and inflammatory response), ulcer area, and colon weight / length ratio were monitored to evaluate the therapeutic effect of the test compounds on the TNBS-induced ulcerative colitis model in SD rats. The results showed that:

[0626] 1) Experimental animals induced by TNBS showed obvious signs of ulcerative colitis, including significant weight loss, significantly increased disease activity score (DAI score), significantly increased colonic adhesion and ulcer score, increased ulcer area of ​​colonic tissue, and significantly increased colon weight / length ratio, indicating that the rat ulcerative colitis model was successfully established and is suitable for this pharmacodynamic study.

[0627] 2) Test compound 1 showed a significant improvement effect on the weight loss response of model animals under the set treatment regimen, and the degree of improvement was statistically different from that of the model control group at the same time (P<0.05), while test compounds 6 and 12 and the positive control drug mesalazine did not show a significant weight improvement effect.

[0628] 3) The tested compounds 1, 6 and 12 showed significant improvement effects on the progression of ulcerative colitis in model animals under the set treatment regimens. Their DAI scores all showed a significant decrease compared with the model control group at the same time point. The therapeutic effect of tested compound 1 was the most obvious. At the same time, the improvement effects of tested compounds 1 and 6 showed obvious dose dependence, while the positive control drug mesalazine did not have a significant effect on the symptoms of ulcerative colitis in model animals.

[0629] 4) The tested compounds 1, 6 and 12 showed significant therapeutic effects on gross colonic injury in model animals under the set treatment regimen. Compared with the control group at the same time, the colonic adhesion score, colonic ulcer and inflammation score and colonic tissue ulcer area all showed significant decreases, and the degree of improvement was statistically significant (P<0.05). The tested compound 1 showed the most significant therapeutic effect, and its therapeutic effect showed a significant dose-dependent effect.

[0630] 5) The tested compounds 1, 6 and 12 showed a significant ameliorative effect on the colonic atrophy process in the model animals under the set treatment regimen. Compared with the control group, the colon weight / length ratio was significantly reduced (P<0.05).

[0631] It is evident that this experiment established a rat model of ulcerative colitis that conforms to clinical pathogenesis. The efficacy evaluation results of this model show that the tested compounds 1, 6 and 12 all have significant therapeutic effects.

[0632] Example 4: Study on the effect of polypeptide compounds on the in vitro proliferation of CaCo-2 cells

[0633] The following cell lines were used to study the in vitro proliferation of polypeptide compounds:

[0634] Table 10 Cell line information

[0635] Serial Number cell lines Cancer types Growth characteristics culture medium 1 CaCo-2 Colorectal cancer Adhesive (MEM + 0.01mM NEAA) + 20% FBS

[0636] Experimental methods:

[0637] The cells will recover and be maintained under the culture conditions shown in Table 10.

[0638] Day 1 (Cell Plating): Collect cells in the logarithmic growth phase, resuspend them, and count and assess cell viability using a cell counter. Dilute the cell suspension according to the plating density requirements, adding 90 μL of cell suspension to each well of a 96-well plate (the cell plating density will be adjusted based on historical data or density optimization experiments). Simultaneously, plating each cell into 3 replicates (T0 plates). Incubate all 96-well plates overnight at 37°C with 5% CO2.

[0639] Day 2 (medium change): Replace the medium with the corresponding serum concentration according to the plate map, 90 μL per well. Day 2: T0 plate reading and test compound processing. Add 10 μL of medium to each well of the T0 plate to bring the total volume to 100 μL. Add 50 μL of medium to each well. Reagents. Mix and vortex for 5 minutes to allow for complete cell lysis. Incubate at room temperature for 10 minutes to stabilize the luminescence signal (Note: temperature, cell density, and edge effects can all cause uneven luminescence signal). Detect the fluorescence signal using an EnVision Multi Label Reader. Prepare the test compounds according to their corresponding molecular weights: C1 = 91.08 μM, C2 = 9.108 μM, and C3 = 0.9108 μM. Prepare 10X working solutions of Cisplatin according to the dilutions shown in Table 11. Add 10 μL of working solution (10X) to each well, for a final volume of 100 μL per well in all plates. Incubate cells at 37°C in a 5% CO2 incubator.

[0640] Day 5, 72 hours after treatment with the test compound (plate reading): Observe the condition of the drug-treated group and the control group under a microscope to see if they are normal. Add 50 μL of CTG reagent to each well. Mix on a shaker for 5 minutes to ensure complete cell lysis. Incubate at room temperature for 10 minutes to stabilize the fluorescence signal. Read the fluorescence signal using an EnVision Multi Label Reader.

[0641] Table 11. Cisplatin concentrations after dilution

[0642] C1-1 C2-1 C3-1 C4-1 C5-1 C6-1 C7-1 C8-1 C9-1 C10-1 Concentration (μM) 1000 333.33 111.11 37.04 12.35 4.12 1.37 0.46 0.15 0 Solution volume (μL) 90 90 90 90 90 90 90 90 90 90 Culture medium volume (μL) 209.7 209.7 209.7 209.7 209.7 209.7 209.7 209.7 209.7 209.7

[0643] Cell viability (%) = Lum 待测化合物 / Lum 溶剂对照 ×100%.

[0644] Experimental results: The statistical results of the proliferation-promoting activities of the selected polypeptide compounds are shown in Table 12:

[0645] Compound numbering Proliferative activity 1 Cell viability >110% 20 / 44 / 45 /

[0646] The experimental system was designed with a test substance concentration gradient of C1 = 91.08 μM, C2 = 9.108 μM, and C3 = 0.9108 μM, with 0.88% FBS as the reference. The experimental results showed that the polypeptide compound 1 of this application has the activity of promoting the proliferation of Caco-2 cells, specifically manifested as an increase in cell viability after intervention.

[0647] In summary, the polypeptide compounds of this invention have short peptide chains, resulting in faster and better absorption. Experimental results show that the polypeptide compounds provided in this application have activity in relieving intestinal distension caused by zebrafish colitis; and exhibit significant therapeutic effects in a rat model of acute inflammatory bowel disease. The polypeptide compounds of this invention have significant efficacy in relieving colitis and can be used to prepare drugs for treating enteritis, especially ulcerative colitis.

[0648] 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 of formula (I) or a physiologically compatible salt thereof, wherein the compound of formula (I) is as follows: H-X a -Glu-Pro-Val-Pro-X b -OH(I) in X a For Lys, Ser-Ser-Glu-Asp-Ile-Lys, or missing; X b It can be Gln-Ala, Ala-Ala-Lys, Gln-Ala-Ala, Glu-Ala-Lys, Gln, or missing.

2. The compound according to claim 1, or a physiologically compatible salt thereof, wherein the compound is selected from: Glu-Pro-Val-Pro (Compound 9); Glu-Pro-Val-Pro-Gln-Ala (compound 43); Glu-Pro-Val-Pro-Ala-Ala-Lys (compound 47); Glu-Pro-Val-Pro-Gln-Ala-Ala (compound 48); Glu-Pro-Val-Pro-Glu-Ala-Lys (compound 55); Lys-Glu-Pro-Val-Pro (compound 85); Glu-Pro-Val-Pro-Gln (compound 86); Lys-Glu-Pro-Val-Pro-Gln (compound 87); Ser-Ser-Glu-Asp-Ile-Lys-Glu-Pro-Val-Pro-Gln (compound 99).

3. Use of the compound of any one of claims 1-2 or a physiologically compatible salt thereof in the preparation of a medicament for treating enteritis.

4. The use according to claim 3, wherein the enteritis includes specific enteritis and nonspecific enteritis; further, specific enteritis includes inflammatory bowel disease, necrotizing enteropathic enteritis and dysbiosis enteritis; further, inflammatory bowel disease includes ulcerative colitis.

5. A pharmaceutical composition comprising a compound of any one of claims 1-2 or a physiologically compatible salt thereof, and a pharmaceutically acceptable excipient.

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