Novel BimBH3 mimic peptide analogue constructed based on nailing and fatty acid acylation double modification strategy as well as preparation method and application of novel BimBH3 mimic peptide analogue

Abstract

The invention discloses a novel BimBH3 mimic peptide analogue constructed on the basis of a nailing and fatty acid acylation dual-modification strategy and application of the novel BimBH3 mimic peptide analogue serving as a PTPN1 inhibitor to development of a long-acting hypoglycemic drug. The structural general formula of the BimBH3 mimic peptide analogue is as shown in a formula I. The structural general formula of the BimBH3 mimic peptide analogue is as shown in a formula I. The invention provides the BimBH3 mimic peptide analogue with a novel structure, and the BimBH3 mimic peptide analogue is constructed by adopting a nailing and fatty acid acylation dual-modification strategy and has the structural general formula as shown in the formula I. The invention further provides a preparation method of the BimBH3 mimic peptide analogue. On the basis of a BimBH3 functional domain core sequence, the novel BimBH3 mimic peptide analogue has target inhibitory activity, metabolic stability and in-vivo action durability through second-site lysine substitution, N-terminal palmitic acid conjugation and fifth, sixth and ninth site glutamic acid side chain mediated synthesis ring nailing modification, and the synthesis ring nailing mode is molecular lactam cyclization. Experiments show that the compound can efficiently inhibit PTPN1 activity, has a long-acting hypoglycemic effect in a type 2 diabetes animal model, and has the potential of being developed into a long-acting antidiabetic drug which is administered once a week. The invention also discloses a solid-phase synthesis preparation method of the compound and application of the compound as a potential PTPN1 inhibitor drug.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to a class of BimBH3 mimic peptide analogs targeting PTPN1 constructed based on a dual modification strategy of pinning and fatty acid acylation, as well as their preparation methods and applications. Background Technology

[0002] Diabetes is a globally prevalent chronic metabolic disease, and its incidence has been steadily increasing in recent years. According to the International Diabetes Federation (IDF), the number of people with diabetes worldwide has exceeded 500 million, and is projected to reach 643 million by 2030 and 783 million by 2045. Based on different clinical manifestations and etiologies, diabetes is mainly divided into three categories: type 1 diabetes, type 2 diabetes, and gestational diabetes mellitus (GDM). Type 2 diabetes (T2DM) accounts for more than 90% of all diabetes cases worldwide, and its main pathological features are insulin resistance and persistent hyperglycemia.

[0003] Currently, antidiabetic drugs are the main treatment for improving hyperglycemia in diabetic patients. Based on the route of administration, commonly used drugs for treating diabetes can be divided into subcutaneous injections (such as insulin and its analogues, GLP-1 receptor agonists) and oral formulations (such as metformin, thiazolidinediones, etc.). However, existing drugs still have certain limitations: for example, the production process of GLP-1 analogues is complex and the synthesis cost is high; some hypoglycemic drugs may be associated with adverse reactions such as hypoglycemia and weight gain.

[0004] Non-receptor protein tyrosine phosphatase 1 (PTPN1, encoded by the PTPN1 gene) is a key member of the protein tyrosine phosphatase (PTP) family. It was first isolated and purified from human placental cells by Tonks et al. in 1988. This enzyme is primarily located in the endoplasmic reticulum of eukaryotic cells. Its C-terminus contains 35 specific amino acid residues, responsible for anchoring to the endoplasmic reticulum and regulating enzyme activity through subcellular localization; the N-terminus is the catalytic domain, extending towards the cytoplasm, where it performs its catalytic function. PTPN1 participates in cell signal transduction and homeostasis by dephosphorylating various signaling proteins. As an important component of the insulin signaling pathway, PTPN1 can remove the phosphate group of phosphorylated insulin receptors, thereby preventing the initiation of subsequent signaling pathways and reducing the ability of glucose transport proteins and glucose uptake. Therefore, it is considered a reverse regulator of insulin signaling and plays a crucial role in this process. Further research has revealed a close relationship between PTPN1 and obesity, type 2 diabetes, and other conditions.

[0005] PTPN1 inhibitors exert their therapeutic effects by specifically inhibiting the enzymatic activity of PTPN1. Currently, the most researched inhibitor types include difluoromethylene phosphates, oxalamidobenzoic acid derivatives, and phenoxyacetic acid derivatives. However, due to the highly conserved and positively charged catalytic pocket of PTPN1, its inhibitor development still faces the following challenges: First, PTPN1 is highly homologous to the catalytic domain of non-receptor protein tyrosine phosphatase 2 (PTPN2, encoded by PTPN2), resulting in low selectivity for small molecule inhibitors; second, PTPN1 is an intracellular protein, and some highly active inhibitors have poor cell membrane permeability, limiting their bioavailability; furthermore, since PTPN1 participates in the regulation of multiple signaling pathways, inhibiting its activity may trigger off-target effects and potential adverse reactions. Therefore, improving selectivity and cell penetration, and reducing off-target risks have become important directions in the current research and development of PTPN1 inhibitors.

[0006] The BH3 region, located within the Bcl-2 family of proteins, is a short α-helical fragment that can promote or inhibit interactions among Bcl-2 family members. Our study reveals that the BimBH3 mimic peptide is a novel PTPN1 inhibitor that effectively alleviates insulin resistance in HepG2 cells, significantly reduces blood glucose levels in a type 2 diabetes animal model, and pathological observations show that the drug has a protective effect on pancreatic cells without causing damage to liver and kidney tissues. In patients with type 2 diabetes, poor glycemic control is mainly due to poor adherence to medication, and there is an urgent clinical need for long-acting anti-type 2 diabetes drugs. Therefore, the development of novel, uniquely mechanismd, and long-acting hypoglycemic drugs based on BimBH3 mimic peptides remains an urgent clinical need, and is of great significance for advancing basic and clinical research in the prevention and treatment of diabetes in my country. (J Med Chem. 2023.02.23;66(4):3030-3044).

[0007] The compounds involved in this invention are derived from the core region of the BimBH3 domain. The N-terminus of the BimBH3 mimic peptide is conjugated with palmitic acid, and a linker-mediated ring-binding on the lysine side chain theoretically extends the duration of drug action in vivo, achieving a once-weekly long-acting hypoglycemic effect. In summary, this type of hypoglycemic peptide targets PTPN1 and has the potential for a once-weekly long-acting hypoglycemic effect, making it worthy of further development into long-acting peptide hypoglycemic drugs. Summary of the Invention

[0008] This invention describes mimetic peptide compounds derived from the core region of the Bim-BH3 domain, their synthesis methods, and applications. The BimBH3 mimetic peptides constructed using the pinning and fatty acid acylation dual modification technique exhibit good PTPN1 and PTPN2 inhibitory activity, resisting degradation by various proteases. These mimetic peptide analogs have a longer duration of pharmacological action, achieving a prolonged anti-type II diabetes effect.

[0009] To achieve the above-mentioned objectives, the present invention adopts the following technical solution: The present invention discloses a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTPN1 with dual modification of stabilization and fatty acid acylation, as well as its preparation method and application.

[0010] The BH3 mimic peptide is derived from the core region of the BimBH3 domain. The N-terminus is modified with palmitic acid, and a linker is used to mediate the cyclization of the lysine side chain. The linking mechanism is intramolecular cyclization.

[0011] Furthermore, the amino acids at the Linker positions are: Ala, D-Ala, Gly, His, Arg, D-Arg, Gly-Gly, Arg-Arg, Gly-Gly-Gly, γ-aminobutyric acid, 5-aminovaleric acid, or 7-aminoheptanoic acid, preferably Gly-Gly and γ-aminobutyric acid.

[0012] Furthermore, the BimBH3 mimic peptide constructed using the novel piercing and fatty acid acylation dual modification technology is linked via intramolecular amide cyclization, with i+3 being the preferred linkage mode.

[0013] Furthermore, the preparation method of the BimBH3 mimic peptide constructed by the novel piercing and fatty acid acylation dual modification technology includes the following steps:

[0014] (1) At room temperature, CTC resin was placed in a polypeptide solid-phase synthesizer and swollen with dichloromethane;

[0015] (2) Add 2-3 times the molar amount of N-Fmoc protected amino acids or palmitic acid and 2.4 times the equivalent amount of HOBt, DIC or Oxyma, DIC, and bubble at 40℃ for 30-60 minutes;

[0016] (3) Add 20% piperidine / DMF mixture to remove Fmoc protecting group for 8+15 min;

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

[0018] (5) Add 2% hydrazine hydrate / DMF solution to remove the protecting group of lysine side chain Dde, add 2 equivalents of Linker and 3 equivalents of Oxyma and DIC, bubble with nitrogen at 40°C for 60 min, add 20% piperidine / DMF mixture to remove Fmoc protecting group, so that the amino group on Linker is exposed.

[0019] (6) Add 0.1 equivalent of tetra(triphenylphosphine)palladium and phenylsilane / DCM solution to remove the protecting group of glutamic acid side chain OAll, so that the carboxyl group of glutamic acid side chain is exposed.

[0020] (7) Add 2.4 equivalents of HATU, HOBt and DIEA, and bubble at 40°C for 30-60 minutes to obtain the complete peptide sequence.

[0021] (8) The product generated in step (7) was added to the lysis buffer, shaken at 40°C for two hours, filtered, and then anhydrous ice-cold ether was added to precipitate the solid. Then, washing and vacuum drying were performed to obtain the crude product of the peptide analog.

[0022] (9) The crude peptide analog was purified by reversed-phase preparative liquid chromatography and freeze-dried to obtain the pure BimBH3 analog peptide constructed by double modification techniques of pinning and fatty acid acylation.

[0023] Further, in step (8), the pyrolysis solution is trifluoroacetic acid: triisopropylsilane: 3,6-dioxa-1,8-octanedithiol: pure water = 92.5:2.5:2.5:2.5.

[0024] Further, in step (8), after pyrolysis and filtration, N2 is blown to remove trifluoroacetic acid.

[0025] The present invention provides a drug or pharmaceutical composition comprising a novel stabilized and fatty acid-acylated dual-modified long-acting hypoglycemic BimBH3 mimic peptide active ingredient targeting PTPN1 as described in claims 1-5, characterized in that the drug or pharmaceutical composition comprises any of the stabilized and fatty acid-acylated dual-modified long-acting hypoglycemic BimBH3 mimic peptides targeting PTPN1 and one or more pharmaceutically acceptable carriers or excipients.

[0026] Furthermore, the BimBH3 mimic peptide analog constructed using the pinning and fatty acid acylation dual modification technology is used to prepare an inhibitor for inhibiting PTPN1 activity.

[0027] Furthermore, the BimBH3 mimic peptide analog constructed using the pinning and fatty acid acylation dual modification technology can be used in the preparation of drugs for the prevention or treatment of diseases targeting PTPN1.

[0028] Furthermore, the diseases include type 2 diabetes, cancer obesity, and Alzheimer's disease, with long-acting anti-type 2 diabetes treatments being preferred;

[0029] Furthermore, the BimBH3 mimic peptide constructed using the pinning and fatty acid acylation dual modification technology is administered orally or by injection.

[0030] Compared with existing technologies, the advantages of this invention are as follows: This invention obtains a novel long-acting hypoglycemic BimBH3 mimic peptide with dual modification of PTPN1 via peptide solid-phase synthesis, involving both stabilization and fatty acid acylation. The peptide analog originates from the core region of the BimBH3 domain, with lysine replaced at position 2 and glutamic acid introduced at positions 5, 6, and 9. Furthermore, a side-chain linker-mediated stabilization ring synthesis strategy is employed. Compared with existing long-acting antidiabetic drugs on the market, this invention has a more unique target, a simpler structure, and relatively lower production costs. Experiments have confirmed that the BimBH3 mimic peptide constructed using the stabilization and fatty acid acylation dual modification technology has high purity and a significant inhibitory effect on PTPN1, making it an excellent PTPN1 inhibitor for application in drug development for PTPN1-targeted diseases such as diabetes, cancer, and obesity. Of particular interest is that this invention, through activity screening, yielded a stabilized and fatty acid acylated dual-modified BimBH3 mimic with a longer pharmacological duration and a once-weekly hypoglycemic effect by inhibiting the PTPN1 target. Therefore, the BimBH3 analogues with dual modification of pinning and fatty acid acylation described in this invention have great development value and commercial prospects in the field of developing long-acting hypoglycemic drugs targeting PTPN1. Attached Figure Description

[0031] Figure 1 This is a solubility effect diagram of the analogues of the present invention;

[0032] Figure 2 This is a characterization diagram of the secondary structure of the far-ultraviolet circular dichroism (CD) spectrum of the analogues of the present invention;

[0033] Figure 3 This is a graph showing the glucose tolerance effect of the analogue G4 of the present invention; Detailed Implementation

[0034] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. Unless otherwise specified, the methods in the following embodiments are conventional methods.

[0035] Example 1

[0036] The specific preparation process of the simulated peptide is as follows:

[0037] (1) At room temperature, CTC resin was placed in a polypeptide solid-phase synthesizer and swollen with dichloromethane;

[0038] (2) Add 2-3 times the molar amount of N-Fmoc to protect amino acids or fatty acids and 2.4 times the equivalent amount of HOBt, DIC or Oxyma, DIC, and bubble at 40℃ for 30-60 minutes.

[0039] (3) Add 20% piperidine / DMF mixture to remove Fmoc protecting group for 8+15 min;

[0040] (4) Repeat steps (2) and (3) until the synthesis of the entire simulated peptide backbone sequence is completed;

[0041] (5) Add 2% hydrazine hydrate / DMF solution to remove the protecting group of lysine side chain Dde, add 2 equivalents of Linker and 3 equivalents of Oxyma and DIC, bubble with nitrogen at 40°C for 60 min, add 20% piperidine / DMF mixture to remove Fmoc protecting group, so that the amino group on Linker is exposed.

[0042] (6) Add 0.1 equivalent of tetra(triphenylphosphine)palladium and phenylsilane / DCM solution to remove the protecting group of glutamic acid side chain OAll, so that the carboxyl group of glutamic acid side chain is exposed.

[0043] (7) Add 2.4 equivalents of HATU, HOBt and DIEA, and bubble at 40°C for 30-60 minutes to obtain the complete peptide sequence.

[0044] (8) The product generated in step (7) was added to the lysis buffer, shaken at 40°C for two hours, filtered, and then anhydrous ice-cold ether was added to precipitate the solid. Then, washing and vacuum drying were performed to obtain the crude product of the peptide analog.

[0045] (9) The crude peptide analog was purified by reversed-phase preparative liquid chromatography and freeze-dried to obtain the pure BimBH3 analog peptide constructed by double modification techniques of pinning and fatty acid acylation.

[0046] The mass spectrometry data and HPLC purity analysis data of the 19 BimBH3 mimic peptide compounds described in this invention are shown in Table 1.

[0047] Table 1. Mass spectrometry and HPLC purity analysis data of BimBH3 mimic peptide analogs.

[0048]

[0049] Example 2: Determination of inhibitory activity of protein tyrosine phospholipase 1B (PTPN1) / T cell protein tyrosine phosphatase (PTPN2)

[0050] In this invention, MES buffer was used as the reaction system, human protein tyrosine phosphatase 1B (PTPN1) was used, p-nitrophenyl phosphate disodium (pNPP) was used as the specific substrate, SM-6 was selected as the lead compound as the positive control and DMSO was used as the negative control, and a screening model based on enzyme reaction rate was established using a 96-well plate as the carrier to search for PTPN1 inhibitors through enzymatic methods.

[0051] The specific implementation steps are as follows: First, add 86 μL of MES buffer (concentration 25 mM, pH 6.5) to a 96-well plate, then add 4 μL of the compound, 10 μL of pNPP (concentration 77 mM), and 100 μL of PTPN1 solution (concentration 50 nM) sequentially, making the total volume 200 μL. Each group is performed in triplicate, using DMSO as a negative control. The plate is shaken on a shaker at 25°C for 1 minute, and the OD values ​​are read on a microplate reader at 1 min, 5 min, and 10 min, respectively. 405 Value, calculate OD 405 The rate of change v (ΔOD / min) was measured. The initial reaction rate of each well showed a linear correlation; the slope of the linear portion of the kinetic curve determined the reaction rate of PTPN1, and enzyme activity was expressed as rate. Data were expressed as x ± SD, and t-tests were used to analyze the data from each group. The formula for calculating the inhibition rate of the compound against PTPN1 was:

[0052] Inhibition rate (%) = (v DMSO -v 样本 ) / v DMSO ×100%

[0053] Among them, v DMSO v 样本 The initial mean response rates of the negative control group and the test compound are represented, respectively. Statistical analysis and processing of inhibition rate data were performed using GraphPad Prism software, and inhibitor-response curves were fitted and IC50 values ​​were calculated. 50 Values. The suppression results are shown in Table 2.

[0054] Table 2. Inhibition results of the tested peptide analogs on PTPN1 / PTPN2 activity

[0055] / : The simulated peptide compound showed an inhibition rate of less than 50% against PTPN1 at an initial screening concentration of 40 μmol / L, so the IC50 value was not determined.

[0056] Example 3: Evaluation of the solubility of the tested simulated peptide analog

[0057] The solubility of a series of peptide analogs was determined using high-performance liquid chromatography (HPLC). The specific method is as described in Example 3: First, the peptide analogs were completely dissolved in dimethyl sulfoxide (DMSO) to prepare gradient concentration solutions, and a standard curve was plotted based on the HPLC peak areas. Then, the peptide analogs were dissolved in pure water to prepare a 1 mg / mL suspension, which was then subjected to ultrasonic disruption and vortexing to promote dissolution before HPLC analysis. The obtained peak areas were substituted into the standard curve to calculate the solubility. The results are as follows: Figure 1 As shown, the solubility of analogs G2 and G4, stapled in the i+3 manner, was improved; the solubility of analog G6, stapled in the i+4 manner, increased only slightly; while the solubility of analog G13, stapled in the i+7 manner, decreased significantly. These results indicate that a dual modification strategy of stapled and fatty acid acylation can effectively improve the solubility of peptide-mimicking compounds, thus providing a feasible approach to enhancing the bioavailability of peptide drugs.

[0058] Example 4: Secondary structure of a peptide analog

[0059] Secondary structure analysis of the BimBH3 analogues obtained through enzyme activity screening was performed using far-ultraviolet circular dichroism (CD) spectroscopy. Results are as follows: Figure 2 As shown, analog G6 exhibits typical α-helix characteristics, while the other mimic peptides show a random coil conformation.

[0060] Example 5: Stability evaluation of DPP-IV enzyme of mimic peptide analogs

[0061] Compounds screened by enzyme activity were subjected to DPP-IV enzyme stability assays. The screened compounds were prepared at specific concentrations and co-incubated with the enzyme. Samples at different time points were quenched with a quenching agent, and the reactions were analyzed by HPLC. The percentage of remaining compound at each time point relative to the origin was determined by the relative peak area. The data were fitted to the exponential decay model in GraphPad Prism software to determine the half-life, which was used to evaluate its stability. The results were then compared with the original compound. The enzyme stability data of the four pinned and fatty acid acylated dual-modified BimBH3 mimic peptides described in this invention are shown in Table 4. As shown in Table 4, the stability of analog G4 was significantly improved. The results indicate that the pinned and fatty acid acylated dual-modification technology significantly improves the antimetabolite stability of the mimic peptides against DPP-IV.

[0062] Table 4. Stability of the tested peptide analogs to DPP IV enzyme degradation

[0063] compound SM6 G2 G4 G6 G13 <![CDATA[Anti-DPP-IV metabolic half-life (t 1 / 2 , h)]]> 3.195 6.576 11.10 6.690 4.238

[0064] Example 6: Enzyme stability evaluation of peptide analogs

[0065] Compounds screened by enzyme activity were subjected to in vitro enzyme stability assays. The screened compounds were prepared at specific concentrations and co-incubated with the enzyme. Samples at different time points were quenched with a quenching agent, and the reactions were analyzed by HPLC. The percentage of remaining compound at each time point relative to the origin was determined by the relative peak area. The data were fitted to an exponential decay model in GraphPadPrism software to determine the half-life, which was used to evaluate stability. The results were then compared with the lead compound. The enzyme stability data of the four pinned and fatty acid acylated dual-modified BimBH3 mimic peptide compounds described in this invention are shown in Table 5.

[0066] Table 5. Stability of the tested peptide analogues to protease hydrolysis

[0067] Example 7: Plasma stability evaluation of BimBH3 mimetic peptide analogs

[0068] The analog peptides were formulated to a specific concentration and then co-incubated with plasma. Samples at different time points were quenched using a quencher (0.2% TFA / CH3CN). After post-processing, the samples were analyzed by analytical HPLC. The percentage of residual compound at each time point relative to the origin was determined by the relative peak area. The half-life was determined by plotting the relationship between the amount of residual peptide and reaction time and fitting the data to an exponential decay model in GraphPad Prism software to evaluate its stability. The results are shown in Table 5, indicating that the analog possesses long-lasting characteristics.

[0069] Table 5. Plasma stability of the tested peptide analogs

[0070]

[0071] Example 8: Evaluation of the glucose tolerance effect of the peptide analogue

[0072] Using G4 as a candidate molecule, an oral glucose tolerance test was conducted in mice to investigate the glucose tolerance of each group after subcutaneous administration. Results are as follows: Figure 3 As shown in Figures A and B, glucose stimulation caused an increase in blood glucose concentration in both the control and experimental groups of mice, reaching a peak approximately one hour after each gavage administration. During the glucose tolerance test (GTT), the increase in blood glucose concentration was significantly inhibited compared to the blank control group. The positive control semaglutide group (0.1 μM / kg) and the high-dose G4 analog group (0.2 μM / kg) showed significant differences compared to the saline control group. Therefore, the experimental results indicate that treatment with the G4 analog peptide can improve glucose tolerance in mice.

[0073] In summary, this invention provides a class of BimBH3 mimic peptides constructed using PTPN1-targeting ligation peptides and fatty acid modification technology, along with their preparation methods and applications. The BimBH3 mimic peptide analogs constructed using this ligation peptide and fatty acid modification technology exhibit good PTPN1 enzyme inhibitory activity and good solubility. The mimic peptide L3 demonstrates excellent resistance to protease hydrolysis, making it suitable for the development of long-acting drugs for the prevention or treatment of type II diabetes. It possesses high development potential and excellent application value.

[0074] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments or make equivalent substitutions for some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions claimed by the present invention.

Claims

1. The present application belongs to the technical field of polypeptide drug design and synthesis, and specifically relates to a kind of BimBH3 Mimetic Peptide Analogue constructed by pinning and fatty acid acylation double modification technology and a preparation method thereof, and the application of the compound as a new non-receptor type protein tyrosine phosphatase 1 (PTPN1, also known as protein tyrosine phosphatase 1B, i.e. PTP1B) inhibitor in the development of long-acting hypoglycemic drugs. The BimBH3 Mimetic Peptide Analogue is characterized in that the analogue is a polypeptide compound with the general formula shown in formula I, or a pharmaceutically acceptable salt, solvate or prodrug thereof; wherein, The N-terminal of the analog is conjugated with a fatty acid chain, and the polypeptide sequence comprises at least one pair of side chain-mediated pegging ring cyclization structure through a linker.

2. The structural general formula of the BimBH3 mimetic peptide compound of formula I.

3. The Bim BH3 mimetic peptide analog of claim 1, wherein The fatty acid chain is palmitic acid; the linker is a natural amino acid, a non-natural amino acid derivative, or a combination thereof, including Ala, D-Ala, Gly, His, Arg, D-Arg, Gly-Gly, Arg-Arg, Gly-Gly-Gly, γ-aminobutyric acid, 5-aminovaleric acid, or 7-aminoheptanoic acid, preferably the linkers are Gly-Gly and γ-aminobutyric acid, respectively.

4. The Bim BH3 mimetic analog of claim 1, wherein The connection mode is intramolecular lactam cyclization, preferably the connection mode is i+3.

5. The method of claim 1, wherein the Bim BH3 mimetic peptide targeting PTPN1 is constructed by the combination of the fatty acid acylation and the dual modification technology, and the preparation method comprises the following steps: (1) At room temperature, place the CTC resin in the polypeptide solid-phase synthesizer, and swell with dichloromethane; (2) add 2 times the resin molar amount of N-Fmoc protected amino acid and 2.4 times the resin molar amount of DIEA in dichloromethane as the solvent, and bubble at room temperature for 60-90 min; (3) use 20% piperidine / dimethylformamide mixture to remove the Fmoc protecting group; (4) add 2 times the resin molar amount of N-Fmoc protected amino acid or palmitic acid and 2.4-4.8 times the resin molar amount of HOBt, DIC or Oxyma, DIC, and bubble at 40°C for 30-60 min; (5) repeat steps (3) and (4) to synthesize the main chain sequence of the mimetic peptide; (6) add 2% hydrazine hydrate / dimethylformamide mixture to remove the Dde protection of the Lys side chain; add 2 times the amount of Linker and three times the amount of Oxyma, DIC, and bubble under nitrogen at 40°C for 60 min; use 20% piperidine / dimethylformamide mixture to remove the Fmoc protecting group to expose the amino group on the Linker; (7) add 0.1 times the amount of tetrakis(triphenylphosphine)palladium, phenylsilane / DCM mixed solution to remove the protection group of the glutamic acid side chain OAll to expose the carboxyl group of the glutamic acid side chain; (8) add 2.4 times the amount of HATU, HOBt, DIEA, and bubble at 40°C for 30-60 min to obtain the complete peptide sequence; (9) add the product obtained in step (8) to the cleavage solution, shake at 40°C for two hours, then filter, and then add anhydrous ethyl ether to precipitate the solid. Then wash and vacuum dry to obtain the crude product of the mimetic peptide analog. The crude product of the polypeptide analog is freeze-dried to convert it into a flocculent or powdery solid, and the BimBH3 mimetic peptide constructed by pegging and fatty acid acylation is obtained.

6. The preparation method of the BimBH3 mimetic peptide according to claim 4, wherein the cleavage solution in step (9) is TFA:TIS:DODT:H2O at 92.5:0.25:0.25:0.

25. Use nitrogen to remove the excess TFA in step (9).

7. The Bim BH3 mimetic analog of claims 1-5, wherein, The drug or pharmaceutical composition comprises any one of the targeting PTPN1 stapled and fatty acid acylated modified Bim BH3 mimetic peptide and one or more pharmaceutically acceptable carriers or excipients.

8. Use of the Bim BH3 mimetic peptide analog according to claims 1-5 in the preparation of a medicament for preventing or treating a disease targeting PTPN1.

9. Use according to claim 7, characterized in that: The disease includes type 2 diabetes, cancer obesity, immune regulation antitumor and Alzheimer's disease, wherein the long-acting anti-type 2 diabetes treatment drug is preferred.

10. Use according to claim 7, characterized in that: The stapled and fatty acid acylated modified Bim BH3 mimetic peptide is administered orally or by injection (subcutaneous injection, intravenous injection).

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