TW201210609A - Glucagon-Like Peptide-2 analogues and their preparation method and use - Google Patents

Abstract

This invention relates to Glucagon-like peptide-2 (GLP-2) analogs and their medical uses, the invention further involves a pharmaceutical composition containing GLP-2 analogues and their pharmaceutical salts. The widely use of the GLP-2 analogues or the pharmaceutical composition containing GLP-2 analogues and their pharmaceutical salts provided by this invention in preventing or treating of gastrointestinal tract-related disorders related with mucosa injury and reducing gastro-intestinal side effects in chemotherapy and radiation therapy.

Description

201210609 6. DISCLOSURE OF THE INVENTION: FIELD OF THE INVENTION The present invention relates to Glucagon Like Peptide _2, 'GLP-2' analogs and their preparation for the prevention and/or treatment of gastrointestinal mucosal damage w Uses for injury-related conditions and drugs that reduce the gastrointestinal side effects of chemotherapy and radiation therapy. [Prior Art] GLP-2 belongs to the pre-glucagon-derived peptide (pr〇giucag〇n_derived __ peptide, PGDP), and is a prohormone convertase (PC) of pr〇giucagon (PG). One of the products of degradation is a single-chain polypeptide consisting of 33 amino acid residues with a molecular weight of 390 Daltons, and its amino acid sequence is highly conserved in mammals [Drucker DJ. Glucagon-like peptide 2 J Clin Endocrinol Metab, 2001 '86: 1758-1774]. GLP-2 is synthesized and released by endocrine cells of the intestinal tract L, and is regulated by feeding, nerve and endocrine factors. It is most important to stimulate the secretion activity of L endocrine cells by eating foods containing carbon-lulu water compounds and fats [Rocca AS] LaGrecaJ, Kalitsky J, Brubaker PL. Monounsaturated fatty acid diets improve glycemic tolerance through increased secretion of glucagon-like peptide-1. Endocrinology, 2001, 142: 1148-55]. The concentration of GLP-2 in the blood circulation was most significant in normal adults after i5 minutes and after i hours. The form of the pro-wife present in GLP-2 in intestinal mucosa and blood circulation is intact GLP-2, GLP-2 (1 -33). The biological half-life of GLP-2(l-33) in human blood circulation 3 94975 201210609 About 7 minutes' its metabolism mainly through renal excretion and dipeptidyl peptidase-4 (DPP- on the intestinal brush border) 4) Hydrolysis of the first two residues of the amine terminal to form inactive GLP-2 (3-33). Rats fed GLP-2 analogues lacking the DDP-4 site of action had significantly better intestinal epithelial growth than rats fed native GLP-2. The clearance of GLP-2 in the blood of rats with bilateral nephrectomy was significantly lower than that of unremoved kidney rats. However, GLP-2 (3-33) has recently been shown to be a competitive antagonist of the GLP-2 receptor (GLP-2R), which inhibits GLP-2C1-33) and nutrient-induced intestinal mucosal growth [Shin ED, Estall] JL, izzo A, Drucker DJ, Brubaker PL. Mucosal adaptation to enteral nutrients is dependent on the physiologic actions of glucagons-like peptide-2 in mice. Gastroenterology, 2005, 128: 1340-53]. The above indicates that the biological activity of GLP-2 is regulated by DDP-4 hydrolysis, its degradation product antagonism and renal clearance. The effects of GLP-2 on the intestinal tract include: specifically stimulating intestinal mucosal growth, enhancing regeneration after intestinal mucosal injury; inhibiting apoptosis of intestinal mucosal epithelial cells and crypt cells; and enhancing basal glucose transporter of intestinal epithelial cells - 2 (glucose transporter-2, GLUT2) performance and glucose

Transport, promote intestinal digestion and absorption; inhibit gastrointestinal motility and gastric acid secretion; enhance intestinal mucosal barrier function; increase intestinal blood supply. The effect of GLP-2 on the intestinal mucosa is essentially independent of age and gender, and has significant effects in intravenous, intramuscular, subcutaneous or intraperitoneal injections. n D'G, Petersen Y, Stoll B, et al. Glucagon-like peptide 2: a nutrient-responsive gut growth factor. J 94975 4 201210609

Nutr, 2001, 131: 709-712]. The GLP-2 receptor (GLP-2R) is a member of the B-type glycosidin-catenin G protein coupled receptor superfamily and is highly tissue-specific. Immunohistochemistry and in situ hybridization have demonstrated that GLP-2R is expressed in human intestinal secretion cells, mouse intestinal neurons, and subepithelial myofibroblasts of rats, mice, and marmosets [Yusta B, Huang L, Munroe D, et al. Enteroendocrine localization of GLP-2 receptor expression in humans and rodents. Gastroenterology, β® 119:744-55 ; Bjerknes M, Cheng H. Modulation of specific intestinal epithelial progenitors by

Enteric neurons. Proc Natl Acad Sci. 2001, 98: 12497-502]. The lack of expression of GLP-2R on intestinal cells suggests that GLP-2R may be indirect in promoting proliferation and cytoprotection. Signaling of GLP-2 in cells is achieved via a specific GLp_2R-mediated cyclic adenosine monophosphate (cAMP) pathway. GLP-2 initiates adenylate cyclization, which causes an increase in intracellular cAMP's latter activation of protein kinase A (PKA), which may then promote gene transcription of cAMP response elements via intracellular ca2+ and/or inositol triphosphate pathways. . GLP-2 can phosphorylate two isomers of ERK-1 and mitogen-activated protein kinase (MAPK) in human intestinal mucosal epithelial cells Caco-2 in vitro via phosphorylated extracellular signal-regulated kinase (MEK) The exosinic acid and tyrosine residues of ERK-2 increase their activity, which significantly promotes Caco-2 cell proliferation. This effect can be achieved by a specific tyrosine kinase inhibitor (Genistein), a phospholipid creatinine 3-kinase (PI3-K) inhibitor αΥ294002), and a mitogen-activated protein number enzyme (10). 94975 5 201210609 Inhibitor (PD098059) blocked. On the other hand, GLP-2R signaling initiation inhibits actinomycete-induced BHK-GLP-2R cells> weeks of death and inhibits the activity of caspase-3 (caspase-3) To reduce the division of poly ADP-ribose polymerase. And regardless of the presence or absence of PKA inhibitors, GLP-2 reduced cycloheximide-induced caspase_3 division. After administration of cycloheximide, GLP- is present in the presence of two kinase inhibitors, the phospholipidinositol-3-kinase-3-kinase (PI3-K) inhibitor LY294002 and the mitogen-activated protein kinase (MAPK) inhibitor PD98054. 2 can also increase cell viability [YustaB, EstallJ, DruckerDJ. Glucagon-like peptide-2 recepter activation engages bad and glycogen synthase kinase-3 in a protei kinase A-dependent manner and ensures apoptosis following inhibition of phosphatidylinositol 3-kinase. J Bi〇i

Chem, 2002, 277:24896-24906; Walsh NA, Yusta B, DaCarabra MP, et al. Glucagon-like peptide-2 recepter activation in the rat intestinal mucosa.

Endocrinology, 2003, 144: 4385-4392], indicating that intracellular signaling of GLP-2 is not completely dependent on the PI3-K and MAPK pathways. Thus, the signaling pathway involved in the GLP-2 gut mechanism is complex including both the cAMP-dependent protein kinase pathway and the tyrosine kinase pathway, the ρΐ3~κ and MAPK pathways. The exact molecular mechanism of GLP-2 action has not yet been fully elucidated. The human trial of GLP-2 intestine protection is still in its infancy and is limited to therapeutic studies in patients with short bowel syndrome. It has been clarified that Glp_2 ear 94975 6 201210609 == role', but whether it can reduce the damage and promote intestinal repair is yet to be confirmed by further clinical trials. Although it is still used in the course of treatment, the biological effects of the examination (4)-2 were found in the study of the guide _, and the study of naked skin injection, ^ GLP-2 significantly promoted the growth of intestinal adhesion, therefore (10) 2 Whether it will hedge the growth of intestinal tumors is still difficult to determine. In the case of short bowel disease!: Ϊ 督 督 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 ; 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督 督the study. It is believed that GLp-2 special intestinal protection & anti-effect has important clinical application value in the prevention and treatment of malabsorption, inflammatory bowel disease, stress gastrointestinal mucosal injury and other diseases. GLP-2 is secreted as a peptide with the following 33 amino acids:

His-Ala-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-ij e Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp,

Leu~Ile-Gln-Thr-Lys-I le-Thr-Asp. GLP-2 rapidly inverts the enzyme DPPiy and cleaves it at the nitrogen position of the 2-position alanine (Ala) to form GLP-2 ' (33) and is inactivated. This rapid enzymatic degradation mechanism of GLP-2 (1-33) in addition to renal clearance results in a half-life of about 7 minutes in the human blood circulation.

The effects of GLP-2 antagonists and their effects on the growth of gastrointestinal tissues are described by Drucker et al. in U.S. Patent No. 5,994,500, which is incorporated herein by reference. US 5,994,500 alters the structure of mammalian GLP-2 via mutations such as substitutions and deletions. GLP-2 analogs and their medical uses are disclosed in US 6, 184, 208, US 5, 789, 379 and US 6,184, 201. The analogs are all obtained by substitution and or deletion of human GLP^7 94975 201210609.

DaCambra et al. (Mark P. DaCambra; Bernardo Yusta; Martin Sumner-Smith; Anna Crivici; Daniel J. Drucker; Patricia L. Brubaker, Structural Determinants for Activity of Glucagon-like Peptide-2, Biochemistry (2000), 39(30) , 8888-8894) describes structural determinants of GLP-2 activity. Examples of such determinants are Phe6 and Thr5,

The Phe6 and Thr5 are believed to be critical for GLP-2 receptor binding and activation.

The GLP-2 analogue [Gly2]GLP-2 is disclosed by Drucker et al. in W097/39031. Two of the alanines were replaced by glycine to render the peptide resistant to DPPIV cleavage. Substitution of alanine is shown to increase the stability and potency of the peptide. This patent application describes how to use Glp-2 analogues against diseases associated with intestinal epithelial mucosal inflammation and injury. These diseases include large-scale small bowel resection, inflammatory bowel disease, chemotherapy-induced mucositis, and ischemic injury.

W002/066511 describes GLp-2 analogs with extended half-life in vivo and their use as medicaments for the treatment of gastrointestinal disorders such as inflammatory bowel disease. W001/14H9 describes the use of h[Gly2]GLP_2ffi for the pretreatment of a drug that inhibits chemotherapy-induced apoptosis and promotes cell survival. SUMMARY OF THE INVENTION An object of the present invention is to provide a GLP-2 analogue ' represented by the following formula (1), and pharmaceutically acceptable salts thereof and derivatives thereof, which include long-chain fats (tetra) and/or silk Derivatives of diols and/or other long-acting modifications of the derivative of the GLP-2 analogue produced by the 94975 8 201210609 or a pharmaceutically acceptable salt of the GLP 2 analogue:

Rl-Zl-Xl-X2-X3-Gly^Ser-Phe-Ser-Asp-Glu-X10-Xll-Thr-Ile-Leu-X15-X16-Leu-Ala-Ala-Arg-Asp-Phe-Ile- X24-X25-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-(Z2)-R2(I)' wherein Z1, Z2, R1 or R2 is absent or is a natural or non-natural amine group An acid, or a peptide sequence of 2 to 33 amino acid units consisting of natural and/or unnatural amino acids and X1, X2, X3, X, XII, X15, X16, X24 or X25 are natural or Non-natural amino acid. Preferably, in the GLP-2 analogue represented by the formula (I) of the present invention, XI is His or D-His. Preferably, the GLP-2 analogue represented by the general formula I of the present invention is

Medium X2 is D-Ala or Gly. Preferably, the GLP-2 analogue X10 represented by the formula I according to the invention is Nle or Met. Preferably, the GLP-2 analogue X15 represented by Formula I of the present invention is Glu or Asp. Preferably, in the GLP-2 analogue represented by the formula I of the present invention, X3 is Glu or Asp, and X3 forms a peptide bond with Gly with an α carboxyl group or a /3 carboxyl group. In particular, the present invention more preferably has the following sequence of GLP-2 analogues: D-His-D-Ala-Asp"'^^y~^^r-f>h6-S6r-Asp_Glu-Nle-Asn -Thr~Iie-Leu-Giu-Asn-Leu-Ala-Ala-Arg-Asp-Phe- 9 94975 201210609

Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp °

Hi s~Gly-Asp-Gly_Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu -Ile-Gln-Thr-Lys-Ile-Thr-Asp.

His-Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp- Leu-Ile-Gln- Thr-Lys-Ile-Thr-Asp ° D-His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Asp-Asn- Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn_Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp ° D_Hi s_Gly_Glu-Gly-Ser-Phe_Ser-Asp_Glu-Met-Asn-Thr-Ile -Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-

Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp. According to the present invention, the present invention also relates to a process for producing a GLP-2 analogue according to the present invention, characterized in that the GLP-2 analogue is prepared by solid state synthesis and purified by reverse phase liquid chromatography. The invention further relates to a pharmaceutical composition comprising a GLP-2 analogue of the invention comprising a pharmaceutically acceptable carrier and/or a cancer chemotherapeutic agent, comprising a therapeutically effective amount. The dosage form of the composition is preferably an injection or a sustained release agent. The present invention also relates to the use of the pharmaceutical composition for the preparation of a medicament for preventing and/or treating a disorder associated with gastrointestinal mucosal damage, wherein the gastrointestinal mucosal damage-related disorder is selected from the group consisting of peptic ulcer, malabsorption syndrome, short bowel syndrome, and back Blind-sand syndrome, inflammatory bowel disease, abdominal spleen diarrhea, tropical 94975 10 201210609 _ type Spoel's abdomen, low-gammaglobulinemia type Spironic thirst, enteritis, enteritis, ulceration Colitis caused by colitis, gastrointestinal damage, or positive gastrointestinal side effects of stagnation, chemotherapy or radiation therapy -: 彳 彳 选 、 、 腹部 腹部 腹部 腹部 腹部 腹部 腹部 腹部 腹部 腹部 腹部 腹部 放射性Inflammation, transmission of wood! · Gastrointestinal mucosal damage caused by colitis or post-infection gastroenteritis, poison or chemotherapeutic agent or DPP_IV (dipeptidyl peptidase_IV) mediated condition. The present invention further relates to the use of the heart 2_filament for the prevention and/or treatment of a medicament for gastrointestinal mucosal injury-related disorders, wherein the gastrointestinal adhesion, damage-related disorder is selected from the group consisting of digestive ulcers, malabsorption syndrome, Short bowel syndrome, ileocecal valve syndrome, inflammatory bowel disease, abdominal spleen diarrhea, .,,, 卞 斯 斯 卢 腹 diarrhea, hypogammaglobulinemia type Spiro diarrhea, enteritis, Localized enteritis, ulcerative colitis, osteoporosis caused by gastrointestinal mucosal damage and/or malnutrition 'Chemical therapy or radiation therapy of gastrointestinal tract J, preferably diarrhea, abdominal painful phlegm, „zone vomiting, radioactive gastrointestinal Gastrointestinal mucosal damage caused by inflammation, infectious gastroenteritis or post-infection gastroenteritis, or a poison or chemotherapeutic agent or DPP-IV (dipeptidyl peptidase-IV) mediated disorder. It can be seen that the present invention relates to A novel GLp 2 analog. The amino acid at one or more positions in the sequence of the glP-2 analog of the present invention is replaced compared to the conventional GLP-2, such that the GLP of the present invention is made -2 analog has a body The improvement of biological activity and the improvement of chemical stability. Specifically, 'the GLP-2 analog of the present invention comprises the GLP-2 sequence of position 1, 2, 3, 1 position, Replacement 11 94975 201210609 and/or replacement combinations at one or more of positions 11 , 15 , 16 , 24 or 25 . The present invention provides GLP-2 with improved chemical stability and or biological activity. Analogs. Accordingly, the present invention provides a GLP-2 analogue represented by Formula I, or a pharmaceutically acceptable salt thereof, or a derivative thereof, etc.:

Rl-Zl-Xl-X2-X3-Gly-Ser-Phe-Ser-Asp-Glu~Xl〇-Xll-Thr-Ile-Leu-X15-X16-Leu-Ala-Ala-Arg-Asp-Phe-Il e-X24-X25-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-(Z2)-R2 (Formula I) 'where Z1, Z2, R1 or R2 are absent 'or natural or non- a natural amino acid, or a peptide sequence of 3 to 2 amino acid units consisting of natural or unnatural amino acids, and again; 1, 2, 3, 乂1〇, 乂1b) 15. X16, X24 or X25 are natural or unnatural amino acids. The GLP-2 analogues of the invention also have "high chemical stability, for example stability against acid hydrolysis, oxidation and deamidation. The substitution of the χ3 position enhances the stability of the GLP-2 analogue of the present invention to acid hydrolysis' Χ10 position substitution can enhance the oxidative stability of the GLP-2 analogue of the present invention. XI1, XI6 and or Χ24 Substitution of one or more positions in the position enhances the stability of the GLP-2 analogs of the invention against the action of the decylamine. Thus, the GLP-2 analogs of the invention are relative to Gly2-GLP-2 Can be shown to degrade against acidic solutions, for deamination and/or for oxygen Enhancement of stability of degradation. The substitution of position X丨5 can improve the yield of the GLP-2 analogues of the present invention by chemical synthesis. GLP-2 analogues are long-chain fatty acids and/or polyethylene. Alcohol or other long-acting modification can prolong its half-life in vivo. Polyethylene glycol modification can effectively mask the analog parent and reduce the recognition of the analog by the in vivo immune system and the endoproteinase 12 94975 201210609 The immunogenicity of the analog and the probability of hydrolysis by the protease in the body are reduced, thereby prolonging the time of GLP-2 analog circulation in the body. In another aspect, the invention relates to a GLP of the invention comprising a carrier mixed with a carrier A composition of the -2 analog or a salt thereof or a derivative thereof, etc. In a preferred embodiment, the composition is a pharmaceutically acceptable composition, the carrier being a pharmaceutically acceptable or pharmaceutically acceptable carrier. The GLP-2 peptide analog of the invention may be a pharmaceutically acceptable acid addition salt of a GLP-2 analogue of the invention.

In another aspect, the invention relates to a GLP-2 analogue of the invention or a salt thereof for use in therapy. In another aspect, the present invention relates to the use of a GLP-2 analogue or a salt thereof or a derivative thereof for the preparation of a medicament for the treatment and/or prevention of a gastrointestinal related disorder, such as treatment of impaired intestinal function, bone mass Neonates with osteoporosis or DPP-1V (dipeptidyl peptidase-1V) mediated disorders. For example, the stomach and bowel-related disorders are selected from the group consisting of peptic ulcer, gastritis, malabsorption syndrome, short bowel syndrome, ileocecal valve syndrome, inflammatory bowel disease, abdominal spleen diarrhea, tropical Spira Diarrhea, hypogammaglobulinemia type Spirafe diarrhea, enteritis, Crohn's disease, ulcerative colitis, diarrhea-related irritable bowel syndrome, small bowel injury or short bowel syndrome. Other conditions that may be treated or otherwise detectable or treatable with GLP-2 analogs of the invention include radiation gastroenteritis, infectious gastroenteritis or post-infection gastroenteritis, and due to toxic agents or other chemical diagnostic agents Caused by intestinal damage. This may require simultaneous or post-administration of GLP-2 analogues with chemotherapy or radiation therapy to reduce the side effects of chemotherapy 13 94975 201210609 or radiation therapy such as diarrhea, abdominal cramps or vomiting, and lower by chemotherapy or Intestinal epithelial structure and functional damage caused by radiation therapy. The invention further relates to a treatment kit comprising a cancer chemotherapeutic drug, a GLP-2 analogue of the invention, and instructions for use. In particular, in a treatment kit, it comprises a cancer chemotherapy drug as a therapeutic agent, a GLP-2 analogue of the present invention, and instructions for use, and each of the drugs or analogs described above is optional and A combination of pharmaceutically acceptable carriers. The therapeutic agents can be packaged separately for separate use or can be provided in the same composition. Accordingly, the present invention also relates to a pharmaceutical composition comprising a cancer chemotherapeutic drug in combination with a pharmaceutically acceptable carrier and a GLP-2 analogue of the invention. Patients with an increased risk of gastrointestinal mucosal formation may desire to select a compound to reduce side effects or eliminate the risk of side effects. For example, when choosing a drug for colon cancer formation in a patient who has colon cancer formation or is at risk of developing colon tumor formation, the choice of a drug that is more selective for the small intestine than the colon is a non-selective drug or a choice for the colon. Drugs that are higher than the small intestine may be more appropriate. In other aspects, the invention also relates to the use of a GLP-2 analogue for the manufacture of a medicament for the treatment and/or prevention of malnutrition. In another aspect, the invention relates to the use of a therapeutically effective amount of a GLP-2 analogue of the invention, or a salt thereof, or a derivative thereof, for use in a medicament for treating a gastrointestinal related disorder in a patient. In another aspect, the present invention relates to a method of treating or preventing a side effect of chemotherapy or radiation therapy in a patient who is in need thereof, the method comprising: using a therapeutically effective amount of the invention described in the present invention a GLp-2 analogue or a salt thereof or a derivative thereof. • The present invention provides a method for treating and preventing the need for poor preparation, which comprises using a therapeutically effective amount of a GLP-2 analogue of the present invention or a salt thereof or the like derivative. . The analog peptide of the present invention is synthesized by the solid phase synthesis technique of a person skilled in the art having the ordinary knowledge in the technical field of the present invention, and the basic principle is as follows: the terminal amino acid of the peptide bond to be synthesized first The mesogenic group is linked to an insoluble polymer resin (solid phase carrier) by a covalent bond structure, and then the amino acid on the carrier is used as an amine group, and the defect is removed by de-ruthenium. Reacts with the (iv) activated ship-based component and joins the long chain. Repeat (condensation - silk - deprotection - washing - lower - round condensation) operation to achieve the desired length of the peptide chain, and finally the peptide chain is cleaved from the resin, I have been purified and treated to obtain the desired polypeptide. The middle control of the condensation and the #deprotection reaction step is a method of detecting the triterpenes. That is, when there is a free amine group on the resin peptide chain, the detection of the trikolidine reagent will show & color/again No coloration when free amine groups (the ninhydrin reagent itself is yellow). Therefore, after the completion of the condensation reaction, if the yellowing/color (the color of the ninhydrin reagent itself) is detected by the knot, the decoupling before the coupling of the amino acid can be carried out after the coupling is completed. Operation, if blue is displayed, it is proved that there are some free amine groups on the peptide bond, and it is necessary to repeat the coupling or change the existing condensed age view - Sansaki is yellow. Principle of detection of agonistic activity of GLP-2 analogues involved in the present invention 15 94975 201210609 is a receptor for GLP-2R coupled to a Gs protein, which, when bound to an agonist, causes an increase in intracellular cAMP concentration. This experiment co-transfects luciferase reporter gene granules regulated by GLP-2R and cAMP response elements in HEK293 cells. When the compound binds to the receptor and activates the receptor, luciferase expression increases. The initiation of GLP-2R by the compound can be obtained by detecting the activity of luciferase. [Embodiment] In order to explain the present invention in more detail, the following examples are given. However, the present invention is not limited thereto, and those skilled in the art can complete the present invention by the technical solution provided by the present invention in combination with the prior art method, and replace the related amino acid to obtain the derivative defined by the general formula I. . Solid phase synthesis of the positive compound Teduglutide of Example 1 The sequence of the positive compound Teduglutide is: H-Hi s-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-

I le-Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Tr p-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH (1) Solid phase Drying and swelling of synthetic resin Weighing Fmoc_Asp(0]tBu)-Wang resin vacuum dried for 24 hours

(0. 5mmol/g) 40g (20mmol) was placed in a 2L bubbling bottle, 400mL DMF swelling resin was added for 30 minutes, DMF solution was removed; (2) Fmoc-Asp(0tBu)-Wang resin removed Fmoc protective orientation Add 200niL2(10) piperidine/DMF solution to the bubbling bottle containing Fmoc-Asp(OtBu)-Wang resin, take it out after 5 minutes, add 20〇raL 20% piperidine/DMF solution, and react at room temperature for 20 minutes. . After the reaction, the resin was washed four times with 16 94975 201210609 DMF 200 mL.

(3) Solid phase synthesis of Teduglutide 1 Condensation Fmoc-Thr(tBu)-〇H Weigh 50mmol Fmoc-Thr(tBu)-0H, add 125mL 0.4M HOBt/DMF to dissolve, then add i50mL0.4MDIC/DCM, in room The reaction was stirred for 10 minutes under temperature; the above solution was added to the resin, and N2 was reacted at room temperature for 4 hours. After the reaction is completed, the reaction solution is withdrawn, followed by DMF, IPA.

And DMF washable resin. 2 Extension of the peptide chain According to the order of the amino acid in the Teduglutide from the carboxy terminus (C-terminus) to the amine terminus (N-terminus)

(H-His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn -T rp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-〇H), the amount of amino acid and condensation reagent is the same as Fmoc-Thr(tBu)-0H, and the protective amino acid is respectively φ Is Fmoc-Thr(tBu)-0H, Fmoc-I le-OH, Fmoc-Lys(Boc)-〇H, Fmoc-Gln(Trt)-〇H, Fmoc-Leu-OH, Fmoc-Trp(Boc)- 0H, Fmoc-Asn(Trt)-0H, Fmoc-Phe-OH, Fmoc-Asp(0tBu)-0H, Fmoc-Arg(Pbf)-OH, Fmoc-Ala-OH, Fmoc-Glu(0tBu)-0H, Fmoc-Met-OH, Fmoc-Ser(tBu)-〇H, Fmoc-Gly-OH and Fmoc-His(Trt)-0H, repeated condensation and deprotection two-step reaction, synthesis. His(Trt)-Ala-Asp (OtBu)-Gly-Ser(tBu)-Phe-Ser(tBu)-A sp(OtBu)-Glu(OtBu)-Met-Asn(Trt)-Thr(tBu)-Ile-Leu-G lu(OtBu) -Asn(Trt)-Leu-Ala-Ala-Arg(Pbf)-Asp(0tBu)-P 17 94975 201210609 he-Ile-Asn(Trt)-Trp(Boc)-Leu-Ile-Gln(Trt)-Thr (tBu) -Lys(Boc)-Ile-Thr(tBu)-Asp(OtBu)-Wang resin. Post-treatment of 3Teduglutide Resin Peptide The resin peptides obtained were washed successively with DMF, IPA and DMF, washed twice with anhydrous diethyl ether and dried in vacuo to give a resin peptide. (4) Preparation of crude Teduglutide peptide The dried resin peptide was added to freshly prepared 10 mL/g resin peptide TFA: ethanedithiol (EDT): TIS: water = 92 · 5: 2. 5: 2. 5 · · 2.5 (volume ratio) of the lysate, reacted at room temperature for 4 hours. After the completion of the reaction, the mixture was filtered, and the resin was washed twice with TFA. The filtrate was collected and combined, and evaporated to 1/3 of the original volume. A large amount of ice-free squirt was added to precipitate Teduglutide, which was centrifuged and vacuum-dried to obtain a white crude peptide. (5) Preparation of Teduglutide by reversed-phase liquid chromatography. 5 g of crude peptide was dissolved in a certain amount of water, filtered through a 0.45 μm membrane and separated by reversed-phase high performance liquid chromatography (RP-HPLC). The mobile phase was A 0. 1 %TFA/H20, B 0. 1% TFA/acetonitrile. Among them, the column was a Denali C-18 column (particle size 8. 3 #m, 5 x 30 cm), the column temperature was 45 degrees', the detection wavelength was 220 nm, and the flow rate was 120 mL/min. The product peak was collected, concentrated to remove most of the acetonitrile, and then lyophilized to give the finished product of Teduglutide 1. 〇g, purity 98.5%, yield 20. 0%. Solid Phase Synthesis of Compound 1 of Example 2 The sequence of Compound 1 is: D-His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Asp-Asn- Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Tr 18 94975 201210609 p-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH, ie in the general formula (I), XI It is D-His, X2 is Gly, X3 is ASP, X10 is Met, X11 is Asn, X15 is Asp, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method was the same as in Example 1. Example 3 Solid phase synthesis of compound 2 The sequence of compound 2 is:

D-His-Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn- Tr p-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH, ie, in the general formula (I), XI is D-His, X2 is Gly, X3 is Glu, X10 is Met, Xll is Asn, X15 is Asp, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method was the same as in Example 1. Example 4 Solid phase synthesis of compound 3 The sequence of compound 3 is: HD-His-Ala-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Th r-Ile-Leu-Glu-Asn- Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH, ie in the general formula (I), XI is D-His X2 is Ala, X3 is Asp, X10 is Met, X11 is Asn, X15 is Glu, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method was the same as in Example 1. Example 5 Solid phase synthesis of compound 4 The sequence of compound 4 is: HD-His-Ala-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Th r-Ile-Leu-Glu-Asn- Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn- 19 94975 201210609

Trp-Leu-I le-Gln-Thr-Lys-I le-Thr-Asp-OH' In the formula (i), XI is D-His, X2 is Ala, X3 is Glu, X10 is Met, Xll It is Asn, X15 is Glu, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method was the same as in Example 1. Example 6 Solid phase synthesis of compound 5 The sequence of compound 5 is: HD-His-D-Ala-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Nle-Asn-Thr-Ile-Leu-Glu-Asn -Leu-Ala-Ala-Arg-Asp-Phe-I le-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH, ie in the general formula (I), XI is D -His, X2 is Ala, X3 is Asp, X10 is Nle, X11 is Asn 'X15 is Glu, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method is the same as in Example 1. Solid Phase Synthesis of Compound 6 of Example 7 The sequence of Compound 6 is: HD-His-D-Ala-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Nle-Asn-Thr-Ile-Leu-Glu-Asn -Leu-Ala-Ala-Arg-Asp-Phe-Ile-As n-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH 'that is, in the general formula (I), XI is D -His, X2 is D-Ala, X3 is Glu, X10 is Nle, Xll 疋Asn ' X15 is Glu ' X16 is Asn, X24 is Asn, X25 is

The Trp ° preparation method was the same as in Example 1. Solid Phase Synthesis of Compound 7 of Example 8 The sequence of Compound 7 is: 20 94975 201210609 H-His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr-Ile-Leu-Glu- Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-A sn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH', ie in the general formula (I), XI is D-His, X2 is Gly, X3 is Asp, ΧΙΟ is Met, X11 is Asn, X15 is Glu, X16 is Asn, X24 is Asn, and X25 is Trp °. The preparation method is the same as in Example 1. Example 9 Solid Phase Synthesis of Compound 8

The sequence of compound 8 is:

His-Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-T hr-I le-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn - Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH' means that in the general formula (I), XI is His, X2 is Gly, X3 is Glu, ΧΙΟ is Met, Xll is Asn, X15 It is Glu, X16 is Asn, X24 is Asn, and X25 is Trp. The preparation method was the same as in Example 1. Solid Phase Synthesis of Compound 9 of Example 10 The sequence of Compound 9 is:

HD-His-Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-A sn-Thr-Ile-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn -Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH' In the formula (I), XI is D-His, X2 is Gly, X3 is Glu, X10 is Met, Xll is Asn, X15 is Glu, X16 is Asn, X24 is Asn, X25 is

The preparation method of Trp ° is the same as that of the example. 21 94975 201210609 The solid phase synthesis of compound 10 of the compound of Example 11 is: H-His-Gly-Asp*(Gly-Ser-Phe-Ser-Asp-Glu-Met-As n-Thr-Ile-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-OH, ie in In the general formula (I), XI is His, X2 is Gly, X3 is Asp, X10 is Met, X11 is Asn, X15 is Glu, X16 is Asn, X24 is Asn, X25 is Trp, and X3 is Asp*: Fmoc -Asp-OtBu. The preparation method is the same as in Example 1. Example 12 agonistic activity assay on GLP-2 receptor (GLP-2R) GLP-2R is a receptor coupled to Gs protein, when the receptor binds to an agonist This results in an increase in intracellular cAMP concentration. This example co-transfects the luciferase reporter plasmid regulated by GLP-2R and cAMP response elements in HEK293 cells, and binds to the receptor when the GLP-2 analog binds to the receptor. At the same time, the expression of luciferase increased. The detection of GLP-2 analogue by GLP-2 analogue was obtained by detecting luciferase activity. The relevant information of each GLP-2 analogue to be tested is shown in Table 1 below. Shown: 22 94975 201210609 Table 1 Molecular weight, mass and final concentration of each GLP-2 analogue for GLP_2R agonistic activity detection Mass (mg) Molecular weight DMSO (ul) Final concentration (mM) Compound 10 ^ 5. 2 3917 132. 7547 10 Compound 4 3 8 3915 97.06258 10 Compound 5 5. 3 3911 135.5152 10 Compound 8 3.8 3931 96.66751 10 Compound 2 6. 6 3916 168. 5393 10 Stone material 3 4.7 3930 119. 5929 10 Compound 9 4.5 3931 114.4747 10 Compound 7 5.7 3916 145.5567 10 Compound 6 5.7 3926 145. 1859 10 ide 5. 3 3901 135.8626 10 Compound 1 6 3901 153.8067 10 GLP-2R and pCRE-Luc plastids were introduced into HEK293 cells by electroporation, and the transfected cells were 40,000. The density of each well/1 〇〇β 1 was seeded into g6 well plates and incubated at 37 ° C for 24 hours. GLP-2 analogues of the invention (each concentration of 3 well replicates) or positive control gIp-2 were added at a concentration gradient and incubated for 5 hours at 37t. Solvent DMS0 was a negative control. Remove 50/^ 丨 medium from each well and add 5 〇 quot 丨 丨 _ 基质 基质 基质 , , ,. The 80/i 1 reaction solution was taken out and added to a white 96-well plate and detected on an Invision plate reader. The EC50 value, 95% confidence, and maximum response rate of the GLP-2 analog are shown in Table 2 below. 23 94975 201210609 Table 2 EC50, 95% confidence and maximum response rate of each GLP-2 analogue GLP-2 or GLP-2 analogue ECsoCnM) 95% confidence (ηΜ) Maximum response rate (8) GLP-2 (positive Control) 0. 03664 0. 01667 to 0. 08054 100 Compound 10 > 100 22.08933 Compound 4 0. 1361 0. 08114 to 0. 2282 99.10504 Compound 5 0. 1195 0. 07614 to 0. 1877 106.4152 Compound 8 0. 08349 0. 04536至0. 1537 86.53016 Compound 2 0. 01105 0. 004686 to 0. 02607 121.9285 Compound 3 0. 2188 0. 1075 to 0. 4454 114.1183 Compound 9 1. 374 0. 6168 to 3. 061 117. 9512 Compound 7 0. 1679 0. 1034 to 0. 2727 103.0608 Compound 6 1. 178 0. 5294 to 2. 623 83.24433 teduglutide 0.04348 0. 008376 to 0. 06585 88.78505 Compound 1 0. 03585 0. 01980 to 0. 06492 90.61841 Both Compounds 1 and 2 showed better GLP receptor agonistic activity. [Simple diagram description] None [Main component symbol description] None 24 94975 201210609 Sequence table

<110>Jiangsu Haosen Pharmaceutical Group Co., Ltd. <120> Glycoglycan peptide-2 analog, and preparation method and use thereof<130> 890031CG <160> 26 <170> Patentln version 3.3 <210 〉

33 PRT artificial sequence <211><212><213><220><221><222><223> GLP-2 analogue positive compound Teduglutide <400>

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Asp Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30 Asp <210> 2 <211> 33 <212> PRT <213> Artificial sequence 201210609 <220><221> His is D-His < 222 > (1) - (1) < 223 > GLP-2 Analog Compound 1 < 400 > 2

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Asp Asn 15 10 15

Leu Ala Ala Arg Asp Phe He Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 3 <211> 33 <212> PRT <213> artificial sequence <220><221> His is D-His <222> (1)..(1) <223>; GLP-2 Analog Compound 2 <400> 3

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Asp Asn 1 5 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30 2 94975 201210609

Asp <210> 4 <211> 33 <212> PRT <213> Artificial sequence <220><221><222><223><400>

His is D-His (1)·.(1) GLP-2 analogue compound 3 4

His Ala Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 1 5 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp

<210> 5 <211> 33 <212> PRT <213> Artificial sequence <220><221> His is D-His <222> (1)..(1) <223> GLP-2 analogue compound 4 <400> 201210609

His Ala Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 6 <211> 33 <212> PRT < 213 > artificial sequence <220><221> His is D-His <222> (1)..(1) <223> GLP-2 Analog Compound 5 <220><221> Ala is D-Ala <222> (2).. (2) <223> GLP-2 Analog Compound 5 <220>221> Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 5 <400>

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys He Thr 20 25 30 4 94975 201210609

Asp <210> 7 <211> 33 <212> PRT < 213 > artificial sequence <220><221> His is D-His < 222 > (1) - (1) < 223 > GLP-2 analogue compound 6 <220><221> Ala is D-Ala <222> (2)..(2) <223> GLP-2 analogue compound 6

<220><221>Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 6 <400〉 7

His Ala Glu Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30 Asp <210> 8 <211> 33 201210609 <212> PRT <213> Manual Sequence <220><221><222><223> GLP-2 analogue compound 7 <400>

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys He Thr 20 25 30

Asp <210> 9 <211> 33 <212> PRT <213> artificial sequence

<220><221><222><223> GLP-2 analogue compound 8 <400>

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu He Gin Thr Lys lie Thr 20 25 30 6 94975 201210609 , * Asp <210> 10 <211> 33 <212> PRT <213>Artificial Sequence<220><221> His is D-His <222> (1)..(1)

<223> GLP-2 analogue compound 9 <400>

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp

<210> 11 <211> 33 <212>PRT < 213 > Artificial sequence <220><221> A peptide bond is formed with a stone carboxyl group <222> (3).. (4 <223> GLP-2 analogue compound 10 <400> 11 7 94975 201210609

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 12 <211> 33 <212> PRT <213> artificial sequence <220><221> His is D-His <222> (1)..(1) <223> GLP-2 Analog Compound 11 <220><221>Xaa is Nle <222> (10)..(10) <223> GLP-2 Analog Compound 11 <400>

His Gly Glu Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp 94975 201210609 > J <210><211><212><213> « 13 33 PRT artificial sequence ' <220> <221> <222><223> His is D-His (1)-(1) GLP-2 analogue compound 12 <220><221><222><223> Ala is D-Ala P)"(2) GLP-2 analogue compound 12 < 220> <221> <222><223> Xaa is Nle (10).. (10) GLP-2 analogue compound 12 <220><221><222><223> Ala Is L-3-(2-naphthyl)-Ala (25).. (25) GLP-2 analogue compound 12 <400> 13

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Gin Ala Leu lie Gin Thr Lys lie Thr 20 25 30 Asp 9 94975 201210609 <210> 14 <211> 33 <212> PRT <213> Manual sequence <220><221> His is D-His <222> (1)..(1) <223> GLP-2 analogue compound 13 <220><221> Ala is D-Ala <222> (2) (2) <223> GLP-2 analogue compound 13 <220><221> Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 13 <220><221> Ala is L-3-(1-naphthyl)-Ala <222> (25).. (25) <223> GLP-2 analogue compound 13 <400>

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 15 10 94975 201210609 Reference <211> 33 <212> PRT <213> Artificial sequence <220><221>His is D-His <222> (1)..(1 <223> GLP-2 analogue compound 14 <220><221> Ala is D-Ala <222> (2)..(2) φφ <223> GLP-2 analogue compound 14 <;220><221> Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 14 <220><221> Ala is L-3-(2' - quinacridyl)-Ala <222> (25).. (25) <223> GLP-2 analogue compound 14 <400>

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Gin Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 16 <211> 33 11 94975 201210609 <212> PRT <213> Artificial sequence <220><221> His is D-His <222> (1)..(1) <223> GLP-2 analogue compound 15 <220><221> Ala is D-Ala <222> (2).. (2) <223> GLP-2 analogue compound 15 <220>;<221> Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 15 <220><221> Ala is L-3-(3'-quinoline喏基)-Ala <222> (25)..(25) <223> GLP-2 analogue compound 15 <400> 16

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 17 <211> 33

<212> PRT 12 94975 201210609 <213> Artificial sequence <220><221> His is D-His <222>(1)..(1) <223> GLP-2 analogue compound 16 <220〉 <221>Ala is D-Ala <222> (2)..(2)

<223> GLP-2 analogue compound 16 <220><221> Xaa is Nle <222>(10)..(10) '<223> GLP-2 analogue compound 16 <220><221>Trp is D-Trp <222> (25)..(25) <223> GLP-2 analogue compound 16 <400>

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn Φ φ 1 5 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 18 <211> 33 <212> PRT <213> Artificial sequence 13 94975 201210609 <220><221> Trp is D-Trp <222> (25)..(25) <223> GLP-2 analogue compound 17 <400> 18

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 19 <211> 33 <212> PRT <213> Artificial sequence <220><221> Ala is L-3-(l-naphthyl)-Ala <222> ) (25) <223> GLP-2 analogue compound 18 <400> 19

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys He Thr 20 25 30 14 94975 201210609

Asp <210> 20 <211> 33 <212> PRT <213> artificial sequence <220><221><222><223><400>

Ala is L-3-(2-naphthyl)-Ala (25)..(25) GLP-2 analogue compound 19 20

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr He Leu Glu Asn 1 5 10 15

Leu Ala Ala Arg Asp Phe lie Gin Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp

<210> 21 <211> 33 <212> PRT <213> Artificial sequence <220><221> Ala is L-3-(l'-quinazolyl)-Ala <222> 25)..(25) <223> GLP_2 analogue compound 20 <400> 21 15 94975 201210609

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 22 <211> 33 <212> PRT < 213 > artificial sequence <220><221> Ala is L-3-(2'-quinaclidyl)-Ala <222> (25).. (25) <223> GLP-2 analogue compound 21 <400>

His Gly Asp Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 1 5 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 23 <211> 33 <212> PRT <213> artificial sequence 16 94975 201210609 <220><221> Trp is D-Trp <222> (25)..(25) <223> GLP-2 analogue compound 22 <400> 23

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Gin Trp Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 24 <211> 33 <212> PRT <213> artificial sequence_ · <220><221> Ala is L-3-(2-naphthyl)-Ala <222> (25)..(25) <223> GLP-2 analogue compound 23 <400〉 24

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30 17 94975 201210609

Asp <210> 25 <211> 33 <212> PRT <213> Artificial sequence <220><221> Ala is L-3-(3'-quinaclidyl)-Ala <222> (25).. (25) <223> GLP-2 analogue compound 24 <400> 25

His Gly Glu Gly Ser Phe Ser Asp Glu Met Asn Thr He Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp <210> 26 <211> 33 <212> PRT <213> Artificial sequence <220><221> His is D-His <222> (1)..(1) 18 94975 201210609 <223> GLP-2 analogue compound 25 <220><221> Ala is D-Ala <222> (2).. (2) <223> GLP-2 analogue compound 25 <220>;<221> Xaa is Nle <222> (10)..(10) <223> GLP-2 analogue compound 25 <220><221> Ala is L-3-(3'-pyridine ())-Ala <222> (25)..(25) <223> GLP-2 analogue compound 25 <400>

His Ala Asp Gly Ser Phe Ser Asp Glu Xaa Asn Thr lie Leu Glu Asn 15 10 15

Leu Ala Ala Arg Asp Phe lie Asn Ala Leu lie Gin Thr Lys lie Thr 20 25 30

Asp 19 94975

Claims (1)

201210609 VII. Patent Application Range: 1. A glucagon-like peptide-2 analog represented by the following general formula (I), a pharmaceutically acceptable salt thereof, and the like, the derivative including a long-chain fatty acid and • / or a derivative of a glycoside-peptide-2 analog produced by polyethylene glycol and/or other long-acting modifications, or a derivative of a pharmaceutically acceptable salt of a glucagon-like peptide-2 analog: Rl-Zl-Xl-X2-X3-Gly-Ser-Phe-Ser-Asp-Glu-X10-XI1-Thr-Ile-Leu-X15-Xl6-Leu-Ala-Ala-Arg-Asp-Phe-Ile- X24-X25-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp-(Z2)-R2(I), wherein Z1, Z2, R1 or R2 is absent or is a natural or unnatural amino acid Or a peptide sequence having 2 to 33 amino acid units consisting of natural and/or unnatural amino acids, and X b X2, X3, X10, XII, X15, X16, X24 or X25 is natural or non- Natural amino acid. 2. The glucagon-like peptide-2 analog of claim 1, wherein XI is His or D-His. 3. A glucagon-like peptide-2 analog according to claim 1 or 2 wherein X2 is D-Ala or Gly. 4. A glucagon-like peptide-2 analog according to claim 1 or 2, wherein X10 is Nle or Met. 5. A glucagon-like peptide-2 analog according to claim 1 or 2 wherein X15 is Glu or Asp. 6. A glucagon-like peptide 2 analog according to claim 1 or 2, wherein X3 is Glu or Asp, and X3 forms a peptide bond with Gly as an alpha carboxyl group or a cold 25 94975 201210609 carboxyl group. 7. The glucagon-like peptide-2 analog according to claim 1, which has the following sequence: D-His-D~Ala-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Nle - Asn-Thr-Ile-Leu-Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp. 8. The glucagon-like peptide-2 analog as described in claim i, which has the following sequence: His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr -Ile-Leu'Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp~Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp. 9. The glucagon-like peptide-2 analog according to claim 1, which has the following sequence: His~Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn-Thr -I le--Leu~Glu-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn'Trp-Leu-Ile-Gln- Thr-Lys-Ile-Thr-Asp. 10. The glucagon-like peptide-2 analog of claim 1, which has the following sequence: ^His-Gly-Asp-Gly-Ser-Phe-Ser-Asp-Glu-Met-Asn Thr -Iie__Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe Ile-Asn~Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp. U. The glucagon-like peptide-2 analog of claim 1, which has the following sequence: D-His-Gly-Glu-Gly-Ser-Phe-Ser-Asp-Glu-Met- 26 94975 201210609 % . Asn-Thr-Ile-Leu-Asp-Asn-Leu-Ala-Ala-Arg-Asp-Phe-Ile-Asn-Trp-Leu-Ile-Gln-Thr-Lys-Ile-Thr-Asp. 12. A method for preparing a glucagon-like peptide-2 analog _ as described in claim i of the patent application, characterized in that the glucosinolate peptide-2 analog is prepared by solid phase synthesis using a reversed phase solution Purified by phase chromatography. 13. A pharmaceutical composition comprising a therapeutically effective amount of a glucagon peptide-2 analogue as described in the scope of claim 2, which comprises a pharmaceutically acceptable carrier and/or a cancer chemotherapeutic agent. 14. The pharmaceutical composition according to claim 3, wherein the dosage form is an injection or a sustained release agent. 15. The glucagon-like peptide 2 analogue described in item 1 of the scope of the patent application is used for the preparation of a medicament for the prevention and/or treatment of gastrointestinal mucosal damage. Selected from digestive tract ulcers, malabsorption syndrome, short bowel syndrome, ileocecal valve syndrome, gastrointestinal bowel disease, abdominal Sprague ventral writing, tropical Sprague's thirst, low-grade t-globulinemia Spiro diarrhea, enteritis, localized enteritis, colitis, gastrointestinal mucosal damage caused by osteoporosis #〇 / or camp ^ disease, gastrointestinal side effects of chemotherapy or radiation therapy, You Lu selected thirsty, abdominal painful phlegm , vomiting, radiation gastritis or post-infection gastroenteritis, poisoning _ _ and ... to produce gastrointestinal membrane damage, or dimorphic enzyme = = gastrointestinal visceral compound composition of the disease, which gastrointestinal, Known injury-related diseases - Thousands of intestinal mucosal m-beat injury related diseases selected from digestive tract ulcers 94975 27 201210609 Ulcers, malabsorption syndrome, short bowel syndrome, ileocecal valve syndrome, inflammatory bowel disease, abdominal Sprague Diarrhea, tropical Spira diarrhea, low Gammaglobulinemia type Spironol diarrhea, enteritis, Crohn's disease, ulcerative colitis, osteoporosis and/or malnutrition caused by gastrointestinal mucosal damage, gastrointestinal side effects of chemotherapy or radiation therapy, preferably diarrhea, Abdominal cramps, vomiting, radiation gastroenteritis, infectious gastroenteritis or post-infection gastroenteritis, gastrointestinal or chemotherapeutic agents caused by gastrointestinal mucosal damage, or dipeptidyl peptidase-IV mediated disorders. 17. A therapeutic kit comprising a cancer chemotherapeutic drug, a glucagon-like peptide-2 analog as described in claim 1 of the patent application, and instructions for use. 28 94975 201210609 IV. Designated representative map: There is no schema in this case (1) The representative representative figure of this case is: (). (2) A brief description of the symbol of the representative figure: 5. If there is a chemical formula in this case, please disclose the chemical formula that best shows the characteristics of the invention: This case does not represent the chemical formula 94975