GLP-2 analogue and application thereof

Abstract

The invention discloses a GLP-2 analogue and application thereof, and belongs to the technical field of biological medicine. The GLP-2 analogue is oligopeptide formed by 1-5 amino acids at the N end of teduglutide, the oligopeptide retains the activity of the GLP-2 analogue, the disease degree of intestinal injury type bowel diseases is effectively improved, and the effect of the GLP-2 analogue is equivalent to that of teduglutide; and since the GLP-2 analogue only comprises five amino acids, the preparation difficulty and cost are remarkably reduced, so that the development cost of clinical medicines is reduced, and the medication cost of the GLP-2 analogue is expected to be reduced.

Description

Technical Field

[0001] The present application belongs to the field of biomedical technology, and specifically relates to GLP-2 analogs and their applications. Background Art

[0002] Glucagon-like peptide-2 (GLP-2) is derived from enteroendocrine L cells and is an intestinal-specific growth factor. It is transcribed and translated from the proglucagon (PG) gene and belongs to the proglucagon derived peptide (PGDP). The main function of GLP-2 is to stimulate the growth of intestinal mucosal cells, inhibit the apoptosis of intestinal mucosal epithelial cells and crypt cells, cause the increase of small intestinal villi, and enhance the intestinal mucosal barrier. GLP-2 can also inhibit gastric emptying and gastric acid secretion, enhance intestinal nutrient absorption and blood flow, and promote intestinal repair after injury. At present, GLP-2 research mainly focuses on gastrointestinal diseases, and is mainly used in the treatment of patients with short bowel syndrome in clinical practice.

[0003] Natural GLP-2 is composed of 33 amino acids, and its amino acid sequence is: HADGSFSDEMNTILDNLAARDFINWLIQTKITD (SEQ ID NO.1). This sequence is highly conserved, and its active circulating form is GLP-2 (1-33). However, natural GLP-2 is easily degraded in the human body. Specifically, the alanine (A) in the second position will be recognized and cut by dipeptidyl peptidase IV (DPP-IV) to generate inactive GLP-2 (3-33), which has a half-life of about 7 minutes in a healthy human body. Therefore, it is necessary to change the molecular structure of GLP-2 to extend its half-life.

[0004] Teduglutide for injection is a GLP-2 analog drug already on the market. It was developed by Takeda and its trade name is Revestive. Its active ingredient is Teduglutide. The difference from natural GLP-2 is that the alanine (A) at the second position of the N-terminus of teduglutide is replaced by glycine (G). This difference does not affect its active function and prevents the hydrolysis of DPP-IV, thereby extending its half-life to about 3 to 5 hours. However, the price of this drug is very expensive. In pediatric patients with short bowel syndrome, although the use of teduglutide for injection can help achieve intestinal autonomy, the annual cost exceeds $400,000.

[0005] In addition, the GLP-2 analogs currently used in clinical research are mostly prepared by chemical synthesis, genetic recombination and other technologies. The preparation process of genetic recombination technology is complex, the synthesis cost is high, and the mechanism and target of action are unclear; while the chemical synthesis technology is difficult to synthesize peptides of >30 amino acids. These factors have greatly limited the widespread application of GLP-2 analogs in clinical research. Summary of the invention

[0006] 1. Problem to be solved

[0007] In order to solve the problem that teduglutide for injection in the prior art is expensive and difficult to prepare because the number of amino acids in the GLP-2 analog teduglutide it contains is greater than 30, the present application provides a GLP-2 analog and its application. The GLP-2 analog is an oligopeptide formed by 1-5 amino acids at the N-terminus of teduglutide, which retains the activity of the GLP-2 analog and effectively improves the disease severity of intestinal injury-type enteropathy. Moreover, since it only includes 5 amino acids, the difficulty and cost of preparation are significantly reduced, thereby reducing the development cost of clinical drugs, and it is expected to reduce the medication cost of GLP-2 analogs.

[0008] 2. Technical solution

[0009] In order to solve the above problems, the technical solutions adopted in this application are as follows:

[0010] The present application provides a GLP-2 analog, which is an oligopeptide formed by 5 amino acids, and the amino acid sequence is: histidine-glycine-aspartic acid-glycine-serine (HGDGS, SEQ ID NO.2). The oligopeptide is a truncated peptide of teduglutide (the amino acid sequence is: HGDGSFSDEMNTILDNLAARDFINWLIQTKITD, SEQ ID NO.3). The inventors unexpectedly found in the study that the oligopeptide retains the activity of the GLP-2 analog and has a biological activity effect that is basically similar to that of teduglutide; while the oligopeptide composed of 4 amino acids (HGDG, SEQ ID NO.4) does not have the activity of the GLP-2 analog and cannot improve inflammatory damage.

[0011] The present application also provides the use of the above-mentioned GLP-2 analogs in the preparation of drugs for treating intestinal diseases caused by intestinal damage.

[0012] Furthermore, the above-mentioned intestinal injury-related intestinal diseases include one or more of hypoxic injury, ulcerative colitis, and sepsis.

[0013] Furthermore, the above-mentioned intestinal injury-type enteropathy includes hypoxic injury.

[0014] Furthermore, the above-mentioned intestinal injury-related intestinal diseases include ulcerative colitis, which manifests as ulcerative colitis after damage by dextran sulfate sodium (DSS).

[0015] Furthermore, the above-mentioned intestinal injury-related enteropathy includes sepsis, which manifests as sepsis after lipopolysaccharide (LPS) injury.

[0016] The present application also provides a pharmaceutical composition, which comprises the above-mentioned GLP-2 analog or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0017] Furthermore, in the above-mentioned pharmaceutical composition, the excipients include: L-histidine, D-mannitol, sodium phosphate monohydrate, disodium hydrogen phosphate heptahydrate and a pH regulator.

[0018] Furthermore, in the above-mentioned pharmaceutical composition, the mass ratio of GLP-2 analogue, L-histidine, D-mannitol, sodium phosphate monohydrate and disodium hydrogen phosphate heptahydrate is (4-6):(3.5-4):(10-20):(0.3-0.8):(3-4).

[0019] Furthermore, in the above-mentioned pharmaceutical composition, the mass ratio of GLP-2 analogue, L-histidine, D-mannitol, sodium phosphate monohydrate and disodium hydrogen phosphate heptahydrate is 5:3.88:15:0.644:3.434.

[0020] Furthermore, the pH adjuster includes sodium hydroxide and / or hydrochloric acid.

[0021] The present application also provides a pharmaceutical preparation, comprising any of the above-mentioned pharmaceutical compositions.

[0022] Furthermore, the above-mentioned pharmaceutical preparation includes an injection preparation in its dosage form.

[0023] Furthermore, the above-mentioned pharmaceutical preparation, the injection preparation includes a subcutaneous injection preparation.

[0024] Furthermore, the above-mentioned pharmaceutical preparation, the injection preparation also includes a diluent for injection, and the diluent for injection includes water for injection.

[0025] Furthermore, in the above-mentioned pharmaceutical preparation, the concentration of the GLP-2 analogue is 5 to 15 mg / mL.

[0026] Furthermore, in the above-mentioned pharmaceutical preparation, the concentration of the GLP-2 analogue is 10 mg / mL.

[0027] Furthermore, the above-mentioned pharmaceutical composition or pharmaceutical preparation can be used to treat intestinal diseases caused by intestinal damage.

[0028] Furthermore, the above-mentioned intestinal injury-related intestinal diseases include one or more of hypoxic injury, ulcerative colitis, and sepsis.

[0029] Furthermore, the above-mentioned intestinal injury-type enteropathy includes hypoxic injury.

[0030] Furthermore, the above-mentioned intestinal injury-related intestinal diseases include ulcerative colitis, which manifests as ulcerative colitis after damage by dextran sulfate sodium (DSS).

[0031] Furthermore, the above-mentioned intestinal injury-related enteropathy includes sepsis, which manifests as sepsis after lipopolysaccharide (LPS) injury.

[0032] The present application also provides the use of the above-mentioned pharmaceutical composition or pharmaceutical preparation in drugs for treating intestinal diseases caused by intestinal damage, which comprises subcutaneous injection of an effective amount of the pharmaceutical composition.

[0033] Furthermore, in the use of the above-mentioned pharmaceutical composition in the treatment of intestinal damage-related enteropathy, the effective amount of the pharmaceutical composition is calculated based on the GLP-2 analogue being 0.05 mg / kg body weight.

[0034] 3. Beneficial effects

[0035] Compared with the prior art, the present application has the following beneficial effects:

[0036] (1) The GLP-2 analogues and their applications provided in the present application are oligopeptides formed by 5 amino acids. When used to treat intestinal damage-related enteropathy or to prepare drugs for treating intestinal damage-related enteropathy, they can promote the repair of intestinal barrier damage and inhibit intestinal cell apoptosis, and their biological activity is close to that of teduglutide.

[0037] (2) The GLP-2 analogs and their applications provided in the present application, compared with natural GLP-2 or teduglutide (both of which have 33 amino acids), the GLP-2 analogs only include 5 amino acids and can be prepared by Fmoc solid phase synthesis. The preparation process is simpler, faster, and less expensive, which can reduce the cost of drug development.

[0038] (3) The GLP-2 analogs and their applications provided in the present application are oligopeptides formed by 5 amino acids. However, as shown in Example 6, the oligopeptide (HGDG) composed of 4 amino acids does not have the biological activity of GLP-2. The difference is only one amino acid, but the effects are significantly different. This also shows that the technical effects of the GLP-2 analogs and their applications provided in the present application are difficult to predict. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 This is a schematic diagram of the principle of preparing GLP-2 analogs using the Fmoc solid phase synthesis method.

[0040] Figure 2 It is the effect of GLP-2 analogues on the proliferation and migration ability of intestinal epithelial cells, where A is the CCK8 test result and B is the cell scratch test result.

[0041] Figure 3 It is the detection result of intracellular glucose content level after intestinal epithelial cell damage.

[0042] Figure 4 It is the result of the cell number and lactate dehydrogenase (LDH) release after intestinal epithelial cell damage.

[0043] Figure 5 This is the result of immunofluorescence detection of connexin ZO-1.

[0044] Figure 6 The results of WB detection of the protein levels of cyclin CDK4 and anti-apoptotic protein BCL2.

[0045] Figure 7 It is the result of detecting the intracellular inflammatory factors IFN-β and IL-1b mRNA levels by PCR.

[0046] Figure 8 This is the result of blood glucose tester detecting blood glucose changes in mice.

[0047] Fig. 9 The results of WB detection of the protein levels of cyclin ZO-1 and Occludin, mouse small intestinal tissue cyclin CDK4 and CDK6, anti-apoptotic protein BCL2, and GLP-2 receptor protein.

[0048] Fig.10 This is the result of HE staining.

[0049] Fig.11 It is the design and process of dextran sodium sulfate (DSS) injury test.

[0050] Fig.12 These are the results of mouse body weight monitoring and DAI scoring.

[0051] Fig.13 This is the result of the mouse colon length test.

[0052] Fig.14 This is the result of HE staining.

[0053] Fig.15 It is the result of PCR detection of different amounts of peptides on the mRNA levels of inflammatory factors TNF-a, IL-1b and IL-6 in intestinal epithelial cells. DETAILED DESCRIPTION

[0054] The present application is further described below in conjunction with specific embodiments.

[0055] It should be noted that the terms such as "upper", "lower", "left", "right", "middle", etc. cited in this specification are only for the convenience of description and are not used to limit the scope of implementation. Changes or adjustments to their relative relationships should be regarded as the scope of implementation of this application without substantially changing the technical content.

[0056] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this application belongs; the term "and / or" used herein includes any and all combinations of one or more of the associated listed items.

[0057] If the specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or conditions recommended by the manufacturer. If the manufacturers of the reagents or instruments are not specified, they are all conventional products that can be purchased commercially.

[0058] As used herein, the term "about" is used to provide flexibility and imprecision associated with a given term, measurement or value. The degree of flexibility for a particular variable can be easily determined by one skilled in the art.

[0059] As used herein, the term "at least one of" is intended to be synonymous with "one or more of." For example, "at least one of A, B, and C" expressly includes only A, only B, only C, and combinations of each thereof.

[0060] Concentration, amount and other numerical data can be presented in range format herein.It should be understood that such range format is only used for convenience and simplicity, and should be flexibly interpreted as not only including the numerical value clearly described as range limit, but also including all single numerical values ​​or sub-ranges contained in the range, just as each numerical value and sub-range are clearly described.For example, the numerical range of about 1 to about 4.5 should be interpreted as not only including the limit value of 1 to about 4.5 clearly described, but also including single numerals (such as 2,3,4) and sub-ranges (such as 1 to 3,2 to 4, etc.).The same principle is applicable to the scope of only narrating a numerical value, such as "less than about 4.5", which should be interpreted as including all the above-mentioned values ​​and ranges.In addition, no matter how the breadth of the described range or feature is, this explanation should be applicable.

[0061] As used in this application, "GLP-2", "natural GLP-2" and "GLP-2-33-aa" all refer to natural GLP-2, whose amino acid sequence is: HADGSFSDEMNTILDNLAARDFINWLIQTKITD (SEQ ID NO.1).

[0062] As used in this application, "teduglutide for injection" is a GLP-2 analog drug already available on the market, developed by Takeda, and its trade name is Revestive.

[0063] As used in this application, "Teduglutide" is the active ingredient of the GLP-2 analog drug "Teduglutide for injection" that has been marketed. It consists of 33 amino acids, and its amino acid sequence is:

[0064] HGDGSFSDEMNTILDNLAARDFINWLIQTKITD (SEQ ID NO. 3).

[0065] As used in this application, "the GLP-2 analogue of the present application" and "GLP-2-5-aa edit" are both the GLP-2 analogue of the present application, and its amino acid sequence is: HGDGS (SEQ ID NO.2).

[0066] As used in this application, "GLP-2-4-aa edit" is an oligopeptide consisting of amino acids 1-4 of teduglutide, and its amino acid sequence is: HGDG (SEQ ID NO. 4).

[0067] As used in this application, "GLP-2-5-aa" refers to the N-terminal 1-5 amino acids of natural GLP-2, and is also a type of GLP-2 analog, and its amino acid sequence is: HADGS (SEQ ID NO.5).

[0068] In the present application, polypeptides or oligopeptides such as "GLP-2", "teduglutide", "GLP-2 analog of the present application", "GLP-2-4-aa edit", and "GLP-2-5-aa" are all dissolved using a biosolubilizer (dimethyl sulfoxide, DMSO).

[0069] Example 1

[0070] This embodiment provides a GLP-2 analog and a method for preparing the same.

[0071] The GLP-2 analog provided in this example has an amino acid sequence of: H-histidine-glycine-aspartic acid-glycine-serine-OH (HGDGS, SEQ ID NO. 1).

[0072] The present application does not limit the preparation method of GLP-2 analogs, which can be prepared by biosynthesis or solid phase synthesis. In this embodiment, an example of solid phase synthesis is provided.

[0073] The Fmoc solid phase synthesis method is used to prepare GLP-2 analogs. Figure 1As shown, on the polymer resin, according to the amino acid sequence of the GLP-2 analog, starting from the carboxyl end, the sequence (condensation → washing → deprotection → neutralization and washing → next round of condensation) is repeated to connect the amino acids into GLP-2 analogs; finally, the peptide chain is cleaved from the resin, and after purification and other treatments, the desired GLP-2 analog is obtained.

[0074] In this embodiment, the GLP-2 analog is prepared and synthesized by Beijing Zhongke Yaguang Biotechnology Co., Ltd., which removes the amino protecting group by an alkaline solvent (piperidine), and then cross-links with the activated carboxyl group of the next amino acid to form a peptide bond. After the synthesis is completed, it is eluted and deprotected by a deprotecting agent (TFA), and purified by reverse HPLC chromatography, and its purification rate is>95%. Compared with synthetic GLP-2 or teduglutide, due to the reduction in the number of amino acids, the prepared GLP-2 oligopeptide product has high purity, high product yield, and low production cost. The GLP-2 analog prepared in this embodiment has a unit price of about 25 yuan / mg, which is about 5% of the unit price of natural GLP-2 (495 yuan / mg) and teduglutide (440 yuan / mg), which greatly reduces the cost.

[0075] Example 2

[0076] This embodiment provides the use of the GLP-2 analog of the present application in the treatment of intestinal damage-related enteropathy.

[0077] In this embodiment, intestinal injury is intestinal epithelial cell hypoxia / reoxygenation (H / R) injury, specifically including:

[0078] Using cells without hypoxia / reoxygenation (control) and cells with hypoxia / reoxygenation but without pretreatment with GLP-2 analogs (H / R) as controls, Ncm460 and Caco-2 cells were pretreated with natural GLP-2 (GLP-2-33-aa) and the GLP-2 analog of the present application (GLP-2-5-aa edit) at different concentrations (100, 200, 400, 800, 1600 nM) for 2 hours, and then the cells were cultured at 37°C with 95% O2 and 5% CO2 for 12 hours, and then the cells were subjected to hypoxia for 12 hours with a mixed anaerobic gas of 1% O2, 5% CO2 and 94% N2.

[0079] The cell viability was detected by CCK8, and the cell migration ability was detected by cell scratching.

[0080] Result analysis:

[0081] CCK8 test results Figure 2As shown in (A), after hypoxia / reoxygenation (H / R) injury, the proliferation ability of Ncm460 and Caco-2 cells was significantly reduced. After a certain concentration of GLP-2-5-aa edit was given, the proliferation ability of intestinal epithelial cells was significantly increased.

[0082] The results of cell scratch assay were as follows Figure 2 As shown in (B), after hypoxia / reoxygenation (H / R) injury, the migration ability of Ncm460 and Caco-2 cells was significantly reduced. After GLP-2-5-aa edit pretreatment, the migration ability of intestinal epithelial cells was significantly increased and higher than that of GLP-2-33-aa pretreatment, but no significant difference was observed.

[0083] At the same time, neither native GLP-2 (GLP-2-33-aa) nor GLP-2-5-aa edit showed toxic effects on intestinal epithelial cells.

[0084] Example 3

[0085] This embodiment provides the use of the GLP-2 analog of the present application in the treatment of intestinal damage-related enteropathy.

[0086] In this embodiment, the intestinal injury is lipopolysaccharide (LPS) injury, which specifically includes:

[0087] After pretreatment of Ncm460 cells with 800 nM GLP-2-5-aa, GLP-2-33-aa (native), GLP-2-5-aa edit and Teduglutide for 2 h, Ncm460 cells were stimulated with 1 μg / ml LPS for 24 h to establish the LPS stimulation model of intestinal epithelial cells.

[0088] ELISA was used to detect the changes in intracellular glucose levels after intestinal epithelial cell damage caused by GLP-2 and GLP-2 analogs; the LDH method and cell number counting method were used to detect the effects on the proliferation ability of intestinal epithelial cells after damage; WB, immunofluorescence and PCR methods were used to detect the effects on cyclins, junction proteins, apoptosis proteins and inflammatory factors IFN-β and IL-1b mRNA levels.

[0089] Result analysis:

[0090] The results of intracellular glucose level detection after intestinal epithelial cell damage are as follows Figure 3 As shown, after pretreatment with GLP-2-5-aaedit and Teduglutide, the intracellular glucose level was basically the same within 24 hours. In contrast, after pretreatment with GLP-2-5-aa and GLP-2-33-aa, the intracellular glucose level decreased significantly at 6 hours and 24 hours.

[0091] The results of cell number and lactate dehydrogenase (LDH) release after intestinal epithelial cell injury are as follows Figure 4 As shown, after pretreatment with GLP-2-5-aaedit and Teduglutide, the number of cells was larger and the release of lactate dehydrogenase was less, indicating that GLP-2-5-aa edit and Teduglutide can inhibit cell apoptosis at basically the same level, but the inhibitory effects are higher than those of GLP-2-5-aa and GLP-2-33-aa.

[0092] The results of immunofluorescence detection of connexin ZO-1 are as follows Figure 5 As shown, after 24 h of LPS stimulation, GLP-2-5-aaedit and Teduglutide pretreatment upregulated the fluorescence intensity of the intercellular junction protein ZO-1.

[0093] The results of WB detection of protein levels of cyclin CDK4 and anti-apoptotic protein BCL2 are as follows Figure 6 As shown, after 24 h of LPS stimulation, GLP-2-5-aa edit and Teduglutide pretreatment upregulated the protein levels of cyclin CDK4 and anti-apoptotic protein BCL2.

[0094] The results of PCR detection of intracellular inflammatory factors IFN-β and IL-1b mRNA levels are as follows Figure 7 As shown, 6 hours after LPS stimulation, the intracellular inflammatory factors IFN-β and IL-1b mRNA levels were significantly downregulated after GLP-2-5-aa edit and Teduglutide pretreatment, and were significantly different from those in the GLP-2-5-aa and GLP-2-33-aa pretreatment groups.

[0095] Example 4

[0096] This embodiment provides the use of the GLP-2 analog of the present application in the treatment of intestinal damage-related enteropathy.

[0097] In this embodiment, the intestinal injury is lipopolysaccharide (LPS) injury, which specifically includes:

[0098] This example provides verification that the GLP-2 analog of the present application has an intestinal protective effect after intestinal damage in mice after lipopolysaccharide (LPS) modeling. Specifically, it includes:

[0099] After intraperitoneal injection of LPS (7.5 mg / kg) in mice, natural GLP-2 and GLP-2 analogs (600 μg / kg) were injected subcutaneously twice a day (9 am and 4 pm) for 5 consecutive days; 1 hour after the second injection of GLP-2 every day, the blood glucose changes of mice were detected by blood glucose meter.

[0100] The effects on the mRNA levels of cyclins, junction proteins, apoptosis proteins, GLP-2 receptors, and inflammatory factors IFN-β and IL-1b in small intestine were detected by WB and PCR. In addition, the effects of GLP-2-5-aa, GLP-2-33-aa, and GLP-2-5-aa edit on intestinal tissue after intestinal injury were observed by HE.

[0101] Result analysis:

[0102] The results of blood glucose tester testing the blood glucose changes of mice are as follows Figure 8 As shown, the blood glucose concentration of mice decreased significantly after LPS stimulation, while the blood glucose concentration of mice increased on the 3rd to 5th day after treatment with GLP-2-5-aa, GLP-2-33-aa, and GLP-2-5-aa edit peptides.

[0103] The results of WB detection of protein levels of cyclin ZO-1 and Occludin, mouse small intestinal tissue cyclin CDK4 and CDK6, anti-apoptotic protein BCL2, and GLP-2 receptor protein are as follows Fig. 9 As shown in the results, after LPS treatment, the expression levels of mouse small intestinal tissue cycle proteins CDK4 and CDK6, junction proteins ZO-1 and Occludin, anti-apoptotic protein BCL2, and GLP-2 receptor protein were significantly decreased; GLP-2-5-aa edit treatment significantly upregulated the LPS-induced decrease in protein levels, and the effect was significantly higher than that of the GLP-2-5-aa and GLP-2-33-aa treatment groups.

[0104] HE staining results Fig.10 As shown, GLP-2-5-aa, GLP-2-33-aa, and GLP-2-5-aa edit treatment can all improve LPS-induced damage to the small intestinal villi of mice.

[0105] Example 5

[0106] This embodiment provides the use of the GLP-2 analog of the present application in the treatment of intestinal damage-related enteropathy.

[0107] In this embodiment, the intestinal injury is dextran sulfate sodium (DSS) injury, which specifically includes:

[0108] Experimental design and process Fig.11 As shown, acute colitis was induced by giving mice 2.5% DSS in their drinking water for 7 days, followed by regular drinking of purified water until the end of the experiment; all drug groups were subcutaneously injected twice daily for 8 consecutive days (all drug injection concentrations were 600 μg / kg). The control group was injected with the same volume of normal saline as the DSS group.

[0109] Body weight and rectal bleeding were monitored daily throughout the experiment. At the end of the experiment, mice were euthanized and colons were collected for analysis of HE injury.

[0110] Result analysis:

[0111] Mouse weight monitoring and DAI score results are as follows Fig.12 As shown, the body weight of mice in the DSS-treated group was significantly reduced and the DAI score was significantly increased; while the GLP-2-5-aa edit-treated group and the positive control group GLP-2-33-aa and Teduglutide groups effectively reversed the symptoms of colitis in mice.

[0112] The results of the mouse colon length test are as follows Fig.13 As shown, compared with the normal group, the colon length of mice in the DSS group was significantly shortened, and the shortening of colon length was alleviated in the GLP-2-5-aa edit treatment group and the GLP-2-33-aa and Teduglutide groups.

[0113] HE test results are as follows Fig.14 As shown, GLP-2-5-aa edit, GLP-2-33aa and Teduglutide peptide treatment can improve the damage of the colon in mice after acute colitis, and there is no significant difference between the GLP-2-5aa edit treatment group and the Teduglutide treatment group.

[0114] Example 6

[0115] This embodiment provides the use of the GLP-2 analogs and other oligopeptides of the present application in the treatment of intestinal diseases such as intestinal damage.

[0116] In this embodiment, the intestinal injury is lipopolysaccharide (LPS) injury, which specifically includes:

[0117] After pretreatment with 800 nM GLP-2-4-aa edit (HGDG), GLP-2-5-aa edit (HGDGS) and Teduglutide for 2 h, RAW cells were stimulated with 1 μg / ml LPS for 24 h to establish the LPS stimulation model of RAW cells (immune cells).

[0118] The PCR method was used to detect the effects of GLP-2 analogues with different peptide chain lengths on the levels of inflammatory factors IFN-βmRNA, TNF-a and IL-1b after RAW cell injury.

[0119] Result analysis:

[0120] like Fig.15The results showed that GLP-2-5-aa edit and Teduglutide peptide treatment improved the inflammatory response of immune cells induced by LPS stimulation, while the GLP-2-4-aa edit treatment group did not improve this inflammatory damage, which also indicates that not any number of truncated Teduglutide will retain its biological activity.

Claims

1. A GLP-2 analogue, characterized in that: The amino acid sequence of the GLP-2 analog is shown in SEQ ID NO.

2.

2. Use of the GLP-2 analogue according to claim 1 in the preparation of a medicament for treating intestinal diseases caused by intestinal damage.

3. The use according to claim 2, characterized in that: The intestinal injury-related enteropathy includes one or more of hypoxic injury, ulcerative colitis, and sepsis.

4. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the GLP-2 analogue or a pharmaceutically acceptable salt thereof according to claim 1, and a pharmaceutically acceptable excipient.

5. A pharmaceutical composition according to claim 4, characterized in that: The auxiliary materials include: L-histidine, D-mannitol, sodium phosphate monohydrate, disodium hydrogen phosphate heptahydrate and a pH regulator.

6. A pharmaceutical composition according to claim 5, characterized in that: The pH adjuster includes sodium hydroxide and / or hydrochloric acid.

7. A pharmaceutical composition according to claim 6, characterized in that: The mass ratio of the GLP-2 analogue, L-histidine, D-mannitol, sodium phosphate monohydrate and disodium hydrogen phosphate heptahydrate is (4-6):(3.5-4):(10-20):(0.3-0.8):(3-4).

8. A pharmaceutical preparation, characterized in that A pharmaceutical composition comprising any one of claims 4-7.

9. A pharmaceutical preparation according to claim 8, characterized in that: The dosage form of the pharmaceutical dosage form is an injection preparation.

10. A pharmaceutical preparation according to claim 9, characterized in that: The injection preparation also includes a diluent for injection, and the concentration of the GLP-2 analog in the injection preparation is 5 to 15 mg / mL.

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