Glycosylation / fatty acid double-site modified BimBH3 analogue as well as preparation method and application thereof

Abstract

The invention discloses a preparation method of a novel BimBH3 mimic peptide analogue modified by glycosylation and fatty acid acylation double sites and application of the BimBH3 mimic peptide analogue as a PTPN1 inhibitor long-acting hypoglycemic drug. The structural general formula of the BimBH3 mimic peptide analogue is as shown in formula I: # imgabs0 # mimic peptide compound is derived from a core region of a BimBH3 structural domain, Fmoc-Lys (Dde)-OH is adopted to replace and modify the second site of a peptide chain, palmitic acid or octadecanedioic acid is adopted to perform conjugation modification on the N end of the peptide chain, sialic acid, galactosyl diacid and fructosyl acid are adopted to perform conjugation modification on the amino group on the side chain of 2-site lysine, and the BimBH3 mimic peptide analogue is obtained through conjugation modification on the amino group on the side chain of 2-site lysine. The glycosylated / fatty acid double-site decoration modified BimBH3 mimic peptide has excellent inhibitory activity on PTPN1, the problem of poor water solubility caused by fatty acylation modification can be remarkably improved after hydrophilic glycosyl is introduced into a molecular structure, and the solubility in an aqueous solution is remarkably improved. The PTPN1 inhibitor has excellent anti-protease and plasma degradation biological stability, is long in half-life period, is a potential long-acting PTPN1 inhibitor, and can be applied to the development of medicines for treating related diseases such as diabetes, cancer, obesity and the like by taking PTPN1 as a target spot.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a class of glycosylation / fatty acid dual-site conjugated modified BimBH3 analogs targeting PTPN1, and a preparation method and application thereof. Background Art

[0002] Diabetes is a global metabolic disease, characterized by "three mores and one less", namely, more drinking, more eating, more urination and weight loss. There are about 529 million diabetics in the world, accounting for about 6.1% of the world's population, and this number is still on the rise. According to the different clinical manifestations of diabetes, diabetes can be divided into three categories: type 1 diabetes, type 2 diabetes and gestational diabetes. Among diabetic patients, more than 90% have type 2 diabetes, which is the result of the combined effects of genes and environment.

[0003] The treatment of diabetes mainly includes diet therapy, exercise therapy and drug therapy. Drug therapy is the first choice for improving hyperglycemia in diabetic patients. Common oral hypoglycemic drugs on the market mainly include biguanides, sulfonylureas, glinides and DPP-IV inhibitors. Injectable drugs mainly include insulin and its analogs and GLP-1 drugs. However, these drugs have some disadvantages. For example, traditional insulin drugs will cause the risk of hypoglycemia in patients. Although GLP-1 long-acting hypoglycemic drugs have excellent long-acting hypoglycemic effects, they have complex structures, high production costs, and are difficult to synthesize, resulting in extremely high drug costs.

[0004] Non-receptor protein tyrosine phosphatase 1 (PTPN1, also known as PTP1B), as a member of protein tyrosine phosphatases (PTPs), was first isolated and purified from human placental cells by Tonks et al. in 1988. PTPN1 is mainly distributed in the endoplasmic reticulum of eukaryotic cells (Silver Spring.18(8),1516-1523, 2010). The C-terminus of PTPN1 has 35 specific amino acid residues, which are used to anchor the endoplasmic reticulum and regulate enzyme activity through intracellular localization of subcellular structures; the N-terminus is the active domain that plays a catalytic role, extending in the direction of the cytoplasm. PTPN1 mainly controls the intracellular signal transduction pathway by regulating tyrosine phosphorylation. PTPN1 can dephosphorylate phosphotyrosine residues on specific proteins, thereby regulating processes such as cell growth, differentiation, metabolism and survival. At present, with the continuous deepening of research on PTPN1, it has been proved to be related to diseases such as obesity, diabetes and cancer. It provides a new therapeutic strategy for the treatment of diseases such as cancer and diabetes.

[0005] PTPN1 is a hot target for type 2 diabetes research at home and abroad in recent years, so the development of PTPN1 inhibitors is extremely important. PTPN1 has a highly conserved and positively charged catalytic pocket, and the development of PTPN1 small molecule inhibitors is hampered by the following three points: First, PTPN1 and PTPN2 have a high degree of homology, resulting in poor selectivity of small molecule inhibitors; second, because PTPN1 is an intracellular protein, some PTPN1 inhibitors with good activity have poor membrane permeability and generally poor in vivo efficacy.

[0006] The BH3 domain is a fragment of the Bcl-2 family protein with high sequence homology. This domain can mediate the interaction between pro-apoptotic and anti-apoptotic Bcl-2 family proteins by binding to the long and hydrophobic groove of the anti-apoptotic family member Bcl-2. In vivo glucose-induced pancreatic β-cell apoptosis requires the participation of Bcl-2 pro-apoptotic proteins Bim, Puma and Bax. Bim is a key mediator of pancreatic β-cell death and hepatic insulin resistance, playing a central role not only in the induction of apoptosis but also in regulating mitochondrial function and cellular metabolism. Bim-deficient model mice have improved hepatic insulin sensitivity, increased lipid oxidation and reduced body fat. In patients with type 2 diabetes, the main reason for poor blood sugar control is poor compliance with drug treatment, and long-acting anti-type 2 diabetes drugs are urgently needed clinically. Therefore, the development of novel structural, unique mechanism of action and long-acting hypoglycemic drugs based on BimBH3 mimetic peptides is still urgently needed clinically, and it is of great significance to promote basic and clinical research in the field of diabetes prevention and treatment in my country.

[0007] Peptide drugs are currently receiving great attention worldwide. Compared with small molecule drugs, peptide drugs have the advantages of high specificity and low toxicity. However, natural peptide drugs have some significant disadvantages, such as short half-life, poor protease stability, poor solubility and low bioavailability, which hinder the widespread clinical application of peptide drugs. There are currently a variety of strategies to extend the half-life of peptide drugs, including fatty acid acylation, pegylation, glycosylation, amino acid site-specific modification, and the development of various sustained-release drug delivery systems and oral preparations. Fatty acid acylation modification usually has a significant impact on the solubility of peptides or small proteins [Diabetologia 38, A192 (1995)], and poor water solubility is the main factor leading to the failure of clinical trials of some candidate drugs with good therapeutic activity in animal experiments. [Bioorg.Med.Chem.56(2022)116614] Glycosylation modification strategies can not only improve the shortcomings of short half-life of molecules such as small molecules, polypeptides and small proteins, but also significantly improve their solubility and improve the bioavailability of polypeptide drugs. The compound involved in the present invention is derived from the core region of the BimBH3 domain, and a fatty acid molecule is coupled to the N-terminus to increase the serum albumin binding rate to prolong the plasma and in vivo half-life, and a sugar group is coupled to the lysine side chain to perform glycosylation / fatty acid dual-site coupling, balance the hydrophilicity / lipophilicity of the compound, and improve the poor water solubility caused by fatty acylation modification. In theory, it can increase the time for the drug to play a role in the body, increase the solubility of the drug, and thus increase the bioavailability of the drug in the body. In summary, this type of hypoglycemic polypeptide is a potential long-acting polypeptide hypoglycemic drug that targets PTPN1, has high solubility and has a long-acting hypoglycemic effect, and has the value of further development. Summary of the invention

[0008] The present invention describes a peptide analog compound derived from the core region of the BimBH3 domain, which is double-modified by fatty acylation and glycosylation, as well as a synthesis method and application thereof. The glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog has a significant inhibitory effect on PTPN1. After the introduction of hydrophilic sugar groups in the molecular structure, the poor water solubility caused by fatty acylation modification can be significantly improved, and the solubility in aqueous solution is significantly improved. At the same time, it still has excellent anti-protease and plasma degradation biological stability, a long half-life, and is a class of potential long-acting PTPN1 inhibitors, which can be used in drug development for related diseases such as diabetes, cancer, obesity, etc. with PTPN1 as the target. It is noteworthy that the present invention has obtained a BimBH3 analog with a long-acting pharmacological action time and high solubility in water through activity screening, and a glycosylation / fatty acid dual-site conjugated modified BimBH3 analog that exerts a long-acting hypoglycemic effect by inhibiting the PTPN1 target.

[0009] To achieve the above-mentioned purpose, the present invention adopts the following technical scheme: The present invention discloses a novel type of long-acting hypoglycemic BimBH3 mimetic peptide targeting PTPN1 with dual-site conjugation modification of glycosylation / fatty acid and its preparation method and application. The general structural formula of the BimBH3 mimetic peptide analog is shown in Formula I:

[0010]

[0011] The BH3 mimetic peptide is derived from the core region of the BimBH3 domain, using lysine to be site-specifically replaced with lysine at position II, conjugated with a small molecule fatty acid at the N-terminus, and coupled with a sugar group on the lysine side chain for glycosylation / fatty acid modification.

[0012] Furthermore, the small molecule fatty acids at the X position are palmitic acid or octadecanedioic acid, and the sugar groups at the Y position are sialic acid, galactaric acid, and fructose acid.

[0013] Furthermore, the preparation method of the novel glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide comprises the following steps:

[0014] (1) Place the CTC resin in a manual peptide solid phase synthesizer at room temperature and swell the resin with DCM;

[0015] (2) Add 2-3 times the molar amount of the resin N-Fmoc protected amino acid or fatty acid and 2.4 times the equivalent of HOBt, DIC or Oxyma, DIC, and react by bubbling at 40°C for 60 minutes;

[0016] (3) adding a piperidine / DMF mixed solution to remove the Fmoc protecting group, and the reaction time is 8 ± 15 min;

[0017] (4) repeating steps (2) and (3) until the synthesis of the entire peptide analog backbone sequence is completed;

[0018] (5) Add 2% hydrazine hydrate / DMF solution to remove the protecting group of the lysine side chain Dde, add 2 equivalents of sugar groups and 3 equivalents of Oxyma and DIC, and bubble nitrogen at 40°C for 60 minutes; the complete peptide sequence is obtained.

[0019] (6) adding the lysate to the product of step (5), shaking on a shaker at 40° C. for 2 h, filtering with absorbent cotton, adding anhydrous icy ether to precipitate the solid, washing, and vacuum drying to obtain a crude product of the peptidomimetic analog;

[0020] (7) The crude peptide analog is purified by reverse phase preparative liquid chromatography and freeze-dried to obtain a pure product of the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog.

[0021] Furthermore, in step (6), the cleavage solution is trifluoroacetic acid: triisopropylsilane: 3,6-dioxa-1,8-octanedithiol: pure water = 92.5:2.5:2.5:2.5.

[0022] Furthermore, in step (6), after the cracking and filtration, N2 is blown to remove trifluoroacetic acid.

[0023] The present invention provides a medicine or pharmaceutical composition containing all the novel glycosylation / fatty acid dual-site conjugated modified long-acting glucose-lowering BimBH3 mimetic peptides targeting PTPN1 as active ingredients described in claims 1-5, characterized in that the medicine or pharmaceutical composition comprises any of the glycosylation / fatty acid dual-site conjugated modified long-acting glucose-lowering BimBH3 mimetic peptides targeting PTPN1 and one or more pharmaceutically acceptable carriers or excipients.

[0024] Furthermore, the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog is used in the preparation of an inhibitor for inhibiting PTPN1 activity;

[0025] Furthermore, the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog is used in the preparation of drugs for preventing or treating diseases targeting PTPN1;

[0026] Furthermore, the diseases include type 2 diabetes, cancer, obesity and Alzheimer's disease, among which long-acting anti-type 2 diabetes therapeutic drugs are preferred. ;

[0027] Furthermore, the administration method of the glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide is oral administration or injection;

[0028] Compared with the prior art, the advantages of the present invention are: the present invention obtains a new type of glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide targeting PTPN1 through a polypeptide solid phase synthesis method, the polypeptide analog is derived from the core region of the BimBH3 domain, the second alanine is site-specifically replaced with lysine, the N-terminus is coupled with a small molecule fatty acid for modification, and the lysine side chain is coupled with a sugar group for modification. Compared with the long-acting hypoglycemic drugs currently on the market, the mimetic peptide has a novel target, a simple structure, and a low preparation cost. The present invention proves through experiments that the glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide has a significant inhibitory effect on PTPN1, and the introduction of hydrophilic sugar groups into the molecular structure can significantly improve the poor water solubility caused by fatty acylation modification, and the solubility in aqueous solution is significantly improved. At the same time, it still has excellent anti-protease and plasma degradation biological stability, a long half-life, and is a potential long-acting PTPN1 inhibitor, which can be used in the development of drugs for related diseases such as diabetes, cancer, obesity, etc. with PTPN1 as the target. What has attracted attention is that the present invention has obtained BimBH3 analogs with long-acting pharmacological action time and high solubility in water through activity screening, and BimBH3 analogs modified by glycosylation / fatty acid dual-site conjugation that exert long-acting hypoglycemic effect by inhibiting PTPN1 target. Therefore, the BimBH3 analogs modified by glycosylation / fatty acid dual-site conjugation described in the present invention have certain development value and commercial prospects in the field of long-acting hypoglycemic drug development targeting PTPN1. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Comparison of solubility data of analogs of the present invention;

[0030] Figure 2 The results of the glucose tolerance test of the analog L3 of the present invention; DETAILED DESCRIPTION

[0031] The technical scheme of the present invention is further described in detail below in conjunction with specific embodiments. The methods in the following embodiments are all conventional methods unless otherwise specified.

[0032] Example 1

[0033] The specific preparation process of the peptide mimetic is as follows:

[0034] (1) Place the CTC resin in a manual peptide solid phase synthesizer at room temperature and swell the resin with DCM;

[0035] (2) Add 2-3 times the molar amount of the resin N-Fmoc protected amino acid or fatty acid and 2.4 times the equivalent of HOBt, DIC or Oxyma, DIC, and react by bubbling at 40°C for 60 minutes;

[0036] (3) adding a piperidine / DMF mixed solution to remove the Fmoc protecting group, and the reaction time is 8 ± 15 min;

[0037] (4) repeating steps (2) and (3) until the synthesis of the entire peptide analog backbone sequence is completed;

[0038] (5) Add 2% hydrazine hydrate / DMF solution to remove the protecting group of the lysine side chain Dde, add 2 equivalents of sugar groups and 3 equivalents of Oxyma and DIC, and bubble nitrogen at 40°C for 60 minutes; the complete peptide sequence is obtained.

[0039] (6) adding the lysate to the product of step (5), shaking on a shaker at 40° C. for 2 h, filtering with absorbent cotton, adding anhydrous icy ether to precipitate the solid, washing, and vacuum drying to obtain a crude product of the peptidomimetic analog;

[0040] (7) The crude peptide analog is purified by reverse phase preparative liquid chromatography and freeze-dried to obtain a pure product of the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog.

[0041] The mass spectrometry data and HPLC purity analysis data of the lead compound and six BimBH3 mimetic peptide compounds described in the present invention are shown in Table 1.

[0042] Table 1. Mass spectrometry data and HPLC purity analysis data of BH3 mimetic peptide analogs

[0043]

[0044] Example 2: Protein tyrosine phosphatase 1B (PTPN1) inhibitory activity assay

[0045] In the present invention, MES buffer was used as the reaction system, human protein tyrosine phosphatase 1B (PTPN1) was utilized, disodium p-nitrophenyl phosphate (pNPP) was used as the specific substrate, the lead compound SM6 was selected as the positive control, and DMSO was used as the negative control. A screening model based on the enzyme reaction rate using a 96-well microplate as a carrier was established, and PTPN1 inhibitors were sought by enzymatic methods.

[0046] The specific implementation method is: using MES buffer system (25mM, pH6.5), adding 86ul MES buffer, 4ul compound, 10μL pNPP (77mM), 100μL PTPN1 solution (50nM) in a 96-well plate, the total reaction volume is 200μL. Each group has 3 parallels, with DMSO as negative control, shaking on a shaker at 25℃ for 1min, and reading the OD of 1min, 5min, and 10min on the microplate reader. 405 Value, calculate OD 405The rate of change v (△OD / min). The reaction rate of each well in the initial stage is linearly correlated. The slope of the linear part of the kinetic curve determines the reaction rate of PTPN1, and the speed represents the enzyme activity. The obtained data are expressed as x±SD, and each group of data is analyzed using t-test. The formula for calculating the inhibition rate of the compound on PTPN1 is:

[0047] Inhibition rate (%) = (v DMSO -v 样本 ) / v DMSO ×100%

[0048] Among them, v DMSO 、v 样本 Represent the initial average reaction rate of the negative control group and the test compound, respectively. GraphPad Prism software was used to perform statistical inhibition rate data statistics and processing, fit the inhibitory dose-effect curve and calculate IC 50 value.

[0049] Table 2. Inhibition results of the tested peptide analogs on PTPN1 activity

[0050]

[0051] Example 3: Solubility evaluation of tested peptidomimetic analogs

[0052] The solubility of the peptide analog was determined by using the determination method in Specific Implementation Case 3. The peptide analog was completely dissolved in DMSO and prepared into a gradient concentration peptide analog solution. A standard curve was drawn based on the HPLC sample peak area. The peptide analog was prepared into a 1 mg / ml suspension with pure water, and the suspension was dissolved by ultrasonic crushing, vortexing, etc., and analyzed by HPLC. The peak area was substituted into the standard curve to obtain the peptide analog solubility. The results are shown in the attached figure. Figure 1 As shown. The solubility of analogs L3 and L6 has been significantly improved, among which the solubility of L3 reaches 487ug / ml and the solubility of L6 is 314ug / ml, which is 10.8 times and 6.9 times higher than that of SM6. The results show that glycosylation modification can significantly improve the solubility of analog peptides, thereby improving the bioavailability of peptide drugs.

[0053] Example 4: Evaluation of DPP-IV Enzyme Stability of Tested Peptide Analogs

[0054] The compounds screened by enzyme activity were subjected to DPP-IV enzyme stability assay, the screened compounds were configured to a certain concentration, incubated with the enzyme, samples at different time points were taken to quench the reaction with a quencher, and analyzed by HPLC after treatment. The remaining compound (%) at each time point relative to the time origin was determined by the relative peak area. The data were fitted with the exponential decay model in the GraphPad Prism software to determine the half-life, which was used to evaluate its stability. Then it was compared with the original compound. The enzyme stability data of the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analog compound described in the present invention are shown in Table 3. As shown in Table 3, the stability of analogs L3 and L6 was significantly improved. The results show that the glycosylation / fatty acid dual-site conjugated modification can significantly improve the anti-metabolic stability of the peptide analog to DPP-IV.

[0055] Table 3. Stability results of tested peptide analogs against DPP-IV enzyme degradation

[0056]

[0057] Example 5: Evaluation of the Enzyme Stability of BimBH3 Peptide Analogs

[0058] The mimetic peptide was subjected to an in vitro enzyme stability assay. The mimetic peptide was configured to a certain concentration and incubated with the enzyme. Samples at different time points were taken to quench the reaction with a quencher, and then analyzed by HPLC after treatment. The percentage of the remaining compound at each time point relative to the time origin was determined by the relative peak area. The data was fitted with the exponential decay model in the GraphPad Prism software to determine the half-life to evaluate its stability. Then it was compared with the original compound. As can be seen from the experimental results, the analogs are more stable than BimBH312-mer, showing great potential as a good long-acting inhibitor of PTP1B. The enzyme stability data of the glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide compound described in the present invention are shown in Table 4.

[0059] Table 4. Proteolytic stability of tested peptidomimetic analogs

[0060]

[0061] Example 6: Evaluation of Plasma Stability of BimBH3 Peptide Analogs

[0062] The analog peptide was formulated into a certain concentration and then incubated with plasma. Samples at different time points were taken to quench the reaction with a quencher (0.2% TFA / CH3CN), and analyzed by analytical HPLC after post-treatment. The percentage of the remaining compound at each time point relative to the time origin was determined by the relative peak area. By plotting the relationship between the remaining peptide amount and the reaction time, the data was fitted with the exponential decay model in the GraphPad Prism software to determine the half-life to evaluate its stability. The results are shown in Table 5, and the analog has long-acting characteristics.

[0063] Table 5. Plasma stability of tested peptide analogs

[0064]

[0065] Example 7: Evaluation of the glucose tolerance effect of BimBH3 mimetic peptide analogs

[0066] Using L3 as a candidate molecule, oral glucose tolerance tests were performed on mice to study the glucose tolerance of each group after subcutaneous administration. The results are shown in the attached Figure 2 As shown in A and B, glucose stimulation caused an increase in blood glucose concentration in the control group and the experimental group mice, reaching a peak at around 60 minutes. During the GTT, compared with the blank control group mice, the increase in blood glucose concentration was significantly inhibited. The positive control semaglutide group (0.1uM / kg) and the analog L3 high-dose group (0.2uM / kg) were significantly different from the saline control group. Therefore, the experimental results show that the treatment of the analog peptide L3 can improve the glucose tolerance of mice.

[0067] In summary, the present invention provides a class of glycosylation / fatty acid dual-site conjugated modified long-acting hypoglycemic BimBH3 mimetic peptides targeting PTPN1 and their preparation methods and applications. The glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide analogs have good PTPN1 enzyme inhibitory activity and good solubility. The mimetic peptide L3 exhibits excellent stability against protease hydrolysis and can be used for the development of long-acting drugs for the prevention or treatment of type 2 diabetes, with high development prospects and excellent application value.

[0068] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for a person skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to replace some of the technical features therein by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions claimed to be protected by the present invention.

Claims

1. The present invention discloses a method for preparing and using a type of BimBH3 mimetic peptide with dual-site modification of glycosylation / fatty acid targeting PTPN1. The general structural formula of the Bim BH3 mimetic peptide analog is shown in Formula I:

2. The preparation method and application of the BimBH3 mimetic peptide targeting PTPN1 glycosylation / fatty acid dual-site coupling to improve solubility according to claim 1. Characterized in that: Palmitic acid or octadecanoic acid is coupled to the nitrogen end of the peptide chain, lysine is used for site-specific substitution modification at position 2 of the peptide chain, and sialic acid, galactaric acid, and fructose acid are introduced into the lysine side chain for conjugation modification. The preferred conjugates are: conjugate X uses palmitic acid, and conjugate Y uses galactaric acid.

3. According to claim 1, the glycosylation / fatty acid dual-site coupled BimBH3 mimetic peptide targeting PTPN1, the preparation method comprising the following steps: (1) Swelling the CTC resin with dichloromethane in a manual peptide solid phase synthesizer at room temperature; (2) adding 2-3 times the molar amount of the resin to the N-Fmoc protected amino acid and 2.4-3.6 times the molar amount of the resin to the DIEA, and bubbling reaction was carried out at room temperature for 60-90 minutes under the condition that the solvent is dichloromethane; (3) adding a 20% piperidine / dimethylformamide mixture to remove the Fmoc protecting group; (4) adding 2 times the molar amount of the resin to the N-Fmoc protected amino acid or fatty acid and 2.4-3.6 times the molar amount of the resin to the HOBt, DIC or HBTU, HOBt and DIEA and bubbling reaction at 40°C for 60-90 minutes; (5) Repeat steps (3) and (4) to complete the synthesis of the polypeptide backbone sequence; (6) The Dde protecting group of the lysine side chain was removed using 2% hydrazine hydrate / DMF solution; (7) Add 3 times the molar amount of the resin sugar group and 3.6 times the molar amount of the resin Oxyma / DIC, and react at 40°C using nitrogen bubbling for 60-90 min to obtain the complete peptide sequence; (8) adding the lysate to the product of step (7), shaking on a shaker at 40° C. for 2 h, filtering, adding anhydrous icy ether to precipitate a solid, washing, and vacuum drying to obtain a crude product of a peptidomimetic analog; The crude BimBH3 mimetic peptide is purified by reverse phase preparative liquid chromatography and freeze-dried to obtain a flocculent or powdery solid, that is, a pure Bim BH3 mimetic peptide modified with dual glycosylation / fatty acid sites.

4. The preparation method according to claim 3, characterized in that: The cleavage solution in step (8) is TFA:TIS:DODT:H2O in a ratio of 92.5:0.25:0.25:0.

25.

5. The preparation method according to claim 3, characterized in that: Excess TFA in step (8) was removed using nitrogen bubbling.

6. A drug or pharmaceutical composition with all the novel glycosylated / fatty acid modified PTPN1-targeted hypoglycemic BimBH3 mimetic peptides described in claims 1 to 5 as active ingredients, characterized in that: The drug or pharmaceutical composition comprises any of the glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptides targeting PTPN1 and one or more pharmaceutically acceptable carriers or excipients.

7. Use of the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analogue according to any one of claims 1 to 5 in improving the solubility of PTPN1 inhibitors.

8. Use of the glycosylation / fatty acid dual-site conjugated modified BimBH3 peptide analogue according to any one of claims 1 to 5 in the preparation of a drug for preventing or treating a disease targeting PTPN1.

9. The use according to claim 8, characterized in that: The diseases include type 2 diabetes, cancer, obesity and Alzheimer's disease, among which long-acting anti-type 2 diabetes therapeutic drugs are preferred.

10. The use according to claim 8, characterized in that: The administration method of the glycosylation / fatty acid dual-site conjugated modified BimBH3 mimetic peptide is oral administration or injection.

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