CN117343145A - Long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B and preparation method and application thereof - Google Patents
Long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B and preparation method and application thereof Download PDFInfo
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- WUJVPODXELZABP-FWJXURDUSA-N trodusquemine Chemical compound C([C@@H]1C[C@H]2O)[C@@H](NCCCNCCCCNCCCN)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@H](C)CC[C@H](C(C)C)OS(O)(=O)=O)[C@@]2(C)CC1 WUJVPODXELZABP-FWJXURDUSA-N 0.000 description 1
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Abstract
The invention discloses a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B, and a preparation method and application thereof. By a means ofThe structural general formula of the Bim BH3 mimic peptide analogue is shown in formula I:wherein n=14, raa is Gly, aib, ser, asp, lys, val, thr, phe, pro, hyp, leu, ile, trp, tyr, cys, met, glu, arg, D-Ala, beta-Ala, D-Ser, D-Leu, D-Pro, D-His and N-CH 3 Ala. The analogue peptide compound is derived from a core region of a Bim-BH3 structural domain, the 2 nd amino acid is modified by adopting natural or unnatural amino acid site-directed substitution, and the N-terminal conjugated small molecule fatty acid is modified, so that the BimBH3 analogue which has longer pharmacological action time and plays a long-acting hypoglycemic effect for one week through inhibiting a PTP1B target is obtained. The compound is prepared by adopting a polypeptide solid-phase synthesis method, and the crude product is purified and freeze-dried to obtain the target compound.
Description
Technical Field
The invention belongs to the field of biological medicine, and particularly relates to a long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B, and a preparation method and application thereof.
Background
Protein tyrosine phosphatase 1B (protein tyrosine phosphatase B, PTP 1B) is a member of the protein tyrosine phosphatase superfamily, which was extracted from human placenta since 1988 and verified to find the earliest PTP family protein, which is mainly distributed on the endoplasmic reticulum of cells, and the crystal structure of which was resolved in 1994. Recent research perspectives have shown that: PTP1B has close relation with the onset and development of type 2 diabetes and obesity, and can be used as a (potential) target for development of anti-tumor and Alzheimer disease medicines. Effects of PTP1B include antagonism with PTK to modulate insulin activity, inhibition of angiogenesis by VEGFR2, tumor antigen presentation, leptin signaling, and the like. PTP1B is a key down-regulating protein in the insulin signaling pathway, and PTP1B inhibitors sensitize insulin-like and insulin-like substances by blocking tyrosine phosphorylation of insulin-stimulated Insulin Receptor (IR), thereby affecting phosphorylation of insulin receptor (IRs-1), lowering blood glucose. At the same time, it can enhance leptin signal, induce fat metabolism level to increase and body weight to decrease. Thus, PTP1B is a popular target for recent studies on T2DM, and a number of candidate compounds have entered preclinical and clinical I, II phase experiments. Some researches find that the over-expression of PTP1B can obviously promote the occurrence and growth of tumors in mice, and the inhibition of the expression of PTP1B by the inhibitor can generate an anti-tumor effect; the mechanism research shows that PTP1B controls the non-mitochondrial oxygen consumption of cells by regulating and controlling the RNF213 gene, so as to promote the survival and growth of tumor cells under the anoxic condition. Accordingly, PTP1B is considered as a target of an antitumor drug. In recent years, PTP1B has also been proposed as a regulatory action in physiological processes related to alzheimer's disease in the central nervous system, and a strategy for inhibiting PTP1B and thus antagonizing adverse physiological processes related to alzheimer's disease regulated by PTP1B has been proposed to develop anti-alzheimer's disease drugs. Therefore, PTP1B has become a potential hot target for anti-diabetic, cancer and Alzheimer's disease drug development, and PTP1B inhibitors are expected to be applied to the development of anti-diabetic drugs targeting PTP 1B. Poor medication compliance in patients with type 2 diabetes (T2 DM) has become one of the major causes of non-ideal glycemic control. The once-weekly antidiabetic agent can significantly improve the convenience, compliance and quality of life of the T2DM patient, and is therefore a clinical need and preference for the T2DM patient.
Currently, inhibitors of PTP1B mainly include PTP1B inhibitors in inorganic small molecule compounds, organic compounds and natural products. To date, there are only three small molecule inhibitors: ertiprotafib, MSI-1436 and JTT-551 enter clinical trials, but they eventually terminate development due to poor efficacy or adverse reactions, poor selectivity and poor cell membrane permeability are the main reasons that hinder the development process of PTP1B small molecule inhibitors; most organic compounds are screened by organic synthesis and combinatorial chemistry methods, firstly, compounds with PTP1B inhibition activity are screened, then substituent groups of the compounds are modified, and finally, a better PTP1B inhibitor is obtained, and the inhibitor has the problems of poor stability, higher charge, over-high lipophilic coefficient and the like, which restrict the formation of drugs; PTP1B inhibitors in natural products are isolated and identified in nature by high throughput screening, and although they have high selectivity and activity, the site of action is not well defined. Therefore, the defects of the existing PTP1B inhibitor molecules are overcome, and the development of novel PTP1B inhibitors which are novel in structure, strong in selectivity, low in toxicity and long in acting is very necessary to meet the urgent requirements of domestic clinic.
The analogue peptide compound is derived from a core region of a Bim-BH3 structural domain, the 2 nd amino acid is subjected to site-directed substitution modification by adopting natural or unnatural amino acid, and the N-terminal is conjugated with a fatty acid small molecule with high serum albumin binding rate, so that the maintenance time of the hypoglycemic effect can be greatly prolonged theoretically, and the long-acting hypoglycemic effect of once-a-week administration can be exerted. In conclusion, the hypoglycemic polypeptide is a potential long-acting polypeptide hypoglycemic drug which targets PTP1B and has long-acting hypoglycemic effect once a week.
Disclosure of Invention
The invention provides a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B, and a preparation method and application thereof. The BimBH3 mimic peptide analogue has good PTP1B inhibition activity, can resist degradation of various proteases and in-vitro plasma, has a remarkably prolonged in-vitro plasma half-life, and can be used for preparing long-acting drug development for preventing or treating related diseases (preferably type 2 diabetes) taking PTP1B as a target.
In order to achieve the aim of the invention, the invention is realized by adopting the following technical scheme: the invention discloses a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B, and a preparation method and application thereof. The structural general formula of the Bim BH3 mimic peptide analogue is shown as formula I:
BimBH3 mimetic peptide compounds of formula I.
The BH3 mimetic peptide is derived from the core region of a Bim-BH3 domain, amino acid 2 is modified by site-directed substitution of natural or unnatural amino acids, and the N-terminus is modified by conjugation with small molecule fatty acids (n=14).
Further, the substituted natural or unnatural amino acids at the Raa position are respectively: gly, aib, ser, asp, lys, val, thr, phe, pro, hyp, leu, ile, trp, tyr, cys, met, glu, arg, D-Ala, beta-Ala, D-Ser, D-Leu, D-Pro, D-His and N-CH 3 -Ala。
Further, the preparation method of the novel BimBH3 mimetic peptide comprises the following steps:
(1) Placing CTC resin in a manual polypeptide solid-phase synthesizer at room temperature, and activating with dichloromethane and dimethylformamide;
(2) Adding a piperidine/dimethylformamide mixed solution to remove Fmoc protecting groups;
(3) Adding 2-3 times of N-Fmoc protected amino acid or fatty acid and 2.4-4.8 times of HOBt, DIC or HOBt, HBTU and DIEA in the molar amount of resin, and carrying out bubbling reaction for 30-60 min at 40 ℃;
(4) Repeating the steps (2) and (3) until the synthesis of the whole mimic peptide sequence is completed;
(5) Adding a lysate into the product obtained in the step (4), oscillating for 2 hours at 40 ℃, filtering, adding absolute ethyl glacial ether to separate out solid, and washing and vacuum drying to obtain a crude product of the analogue;
(6) And purifying the crude peptide analogue product by using reverse phase preparation liquid chromatography, collecting a target peak mobile phase solution, removing acetonitrile, and freeze-drying to obtain flocculent or powdery solid, thus obtaining the BimBH3 simulated peptide analogue pure product.
Further, the lysate comprises triisopropylsilane, 3, 6-dioxa-1, 8-octanedithiol, water and trifluoroacetic acid.
Further, the step (5) is carried out after filtering, and N is blown 2 Excess trifluoroacetic acid is removed.
The invention also provides a medicament or a pharmaceutical composition taking the novel targeted PTP1B long-acting hypoglycemic BimBH3 mimic peptide as an active ingredient, which is characterized in that the medicament or the pharmaceutical composition comprises any novel targeted PTP1B long-acting hypoglycemic BimBH3 mimic peptide and one or more pharmaceutically acceptable carriers or excipients.
Further, the BimBH3 mimetic peptide analogues are used for preparing inhibitors for inhibiting PTP1B activity;
further, the BimBH3 mimic peptide analogue is applied to the preparation of a medicament for preventing or treating diseases taking PTP1B as a target point;
further, the diseases include diabetes, cancer and alzheimer's disease, with the preferred indication being type 2 diabetes (once a week long-acting therapeutic);
furthermore, the medicine or the medicine composition taking the BimBH3 mimic peptide as an active ingredient is orally taken or injected.
Compared with the prior art, the invention has the advantages that: the invention obtains a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B by a polypeptide solid-phase synthesis method, the polypeptide compound is derived from a core region of a Bim-BH3 structural domain, the 2 nd amino acid is modified by adopting natural or unnatural amino acid site-directed substitution, and the N-terminal conjugation is modified by small molecular fatty acid. Experiments prove that the BimBH3 mimic peptide has remarkable inhibition effect on protein tyrosine phosphatase 1B (PTP 1B), and the obtained mimic peptide has high purity, can be used as an excellent PTP1B inhibitor, and can be applied to drug development of related diseases taking PTP1B as a target, such as diabetes, cancer, alzheimer disease and the like. Particularly, the BimBH3 analogues with longer pharmacological action time and long-acting hypoglycemic effect once a week through inhibiting PTP1B targets are obtained through activity screening. Therefore, the BimBH3 analogue has great development value and excellent commercialization prospect in the field of long-acting hypoglycemic drug development of targeted PTP 1B.
Drawings
FIG. 1 shows the results of the once-a-week hypoglycemic effect of the BimBH3 mimetic peptide compound of the present invention.
Detailed Description
The technical scheme of the invention is further described in detail below with reference to specific embodiments. The methods in the following examples are conventional methods unless otherwise specified.
Example 1
The synthetic route taking the compound DFY-1 as an example is specifically prepared as follows:
(1) Resin activation: the corresponding amount of CTC resin is weighed and placed in a manual polypeptide solid-phase synthesizer, and 5ml of DCM is added for swelling and activation for 1h.
(2) Connection Phe (F): DMF was washed 3 times, fmoc-Phe-OH in 3 times the resin molar amount, DIEA in 3.6 times the resin molar amount were added, dissolved in 10ml DCM, and the reaction was bubbled at 40℃for 1h, and DMF was washed 4 times. 5ml of blocking solution (DCM: meOH: DIEA=17:2:1) blocked the resin 3 times, 10min each time, 4 washes with DMF. Fmoc protecting groups were removed twice (8min+15min) by addition of 20% piperidine DMF, washed 4 times with 5ml DMF and assayed by Kaiser's reagent.
(3) Connection Glu (E): washing 3 times with DMF, adding Fmoc-Glu (OtBu) -OH with 2 times of resin molar quantity, HOBt and DIC with 2.4 times of resin molar quantity respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times with DMF, adding 20% piperidine DMF to remove Fmoc protecting group twice (8 min+15 min), washing 4 times with 5ml of DMF, and detecting by Kaiser's reagent.
(4) Connection Asp (D): washing 3 times with DMF, adding Fmoc-Asp (OtBu) -OH in an amount which is 2 times the molar amount of the resin, HOBt and DIC in an amount which is 2.4 times the molar amount of the resin respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times with DMF, removing Fmoc protecting groups by adding 20% piperidine DMF twice (8 min+15 min), washing 4 times with 5ml of DMF, and detecting by Kaiser's reagent.
(5) Connecting Gly (G): washing 3 times by DMF, adding Fmoc-Gly-OH with 2 times of resin mole amount, HOBt with 2.4 times of resin mole amount and DIC respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times by DMF, adding 20% piperidine DMF to remove Fmoc protecting groups twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(6) Connection Ile (I): washing 3 times by DMF, adding 2 times of Fmoc-Ile-OH with the molar weight of resin, 2.4 times of HOBt with the molar weight of resin and DIC respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times by DMF, adding 20% piperidine DMF to remove Fmoc protecting groups twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(7) Ligation Arg (R): washing 3 times with DMF, adding Fmoc-Arg (Mtr) -OH with 2 times of resin mole amount, HOBt and DIC with 2.4 times of resin mole amount respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times with DMF, adding 20% piperidine DMF to remove Fmoc protecting group twice (8 min+15 min), washing 4 times with 5ml of DMF, and detecting by Kaiser's reagent.
(8) Ligation Arg (R): washing 3 times by DMF, adding Fmoc-Arg (Mtr) -OH with 2 times of resin molar quantity, HOBt and DIC with 2.4 times of resin molar quantity respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 1h, washing 4 times by DMF, adding 20% piperidine DMF to remove Fmoc protecting group twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(9) Connection Leu (L): washing 3 times by DMF, adding 2 times of Fmoc-Leu-OH, 2.4 times of HBTU, HOBt and DIEA respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times by DMF, adding 20% of piperidine DMF to remove Fmoc protecting groups twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(10) Connection Glu (E): washing 3 times with DMF, adding Fmoc-Glu (OtBu) -OH with 2 times of resin molar quantity, HOBt and DIC with 2.4 times of resin molar quantity respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times with DMF, adding 20% piperidine DMF to remove Fmoc protecting group twice (8 min+15 min), washing 4 times with 5ml of DMF, and detecting by Kaiser's reagent.
(11) Connection Gln (Q): washing 3 times with DMF, adding Fmoc-Glu (OtBu) -OH with 2 times of resin molar quantity, HOBt and DIC with 2.4 times of resin molar quantity respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times with DMF, adding 20% piperidine DMF to remove Fmoc protecting group twice (8 min+15 min), washing 4 times with 5ml of DMF, and detecting by Kaiser's reagent.
(12) Connecting Gly (G): washing 3 times by DMF, adding Fmoc-Ala-OH with 2 times of resin mole amount, HOBt with 2.4 times of resin mole amount and DIC respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times by DMF, adding 20% piperidine DMF to remove Fmoc protecting groups twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(13) Connection Ile (I): washing 3 times by DMF, adding Fmoc-Ala-OH with 2 times of resin mole amount, HOBt with 2.4 times of resin mole amount and DIC respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 30min, washing 4 times by DMF, adding 20% piperidine DMF to remove Fmoc protecting groups twice (8 min+15 min), washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(14) Linking palmitic acid: washing 3 times by DMF, adding 4 times of palmitic acid, 4.8 times of HBTU, HOBt and DIEA respectively, dissolving in 10ml of DMF, bubbling at 40 ℃ for 1h, washing 4 times by 5ml of DMF, and detecting by Kaiser's reagent.
(15) 5ml DCM was washed 4 times, 5ml DMF was washed 4 times, 5ml MeOH was washed 4 times and drained.
(16) Cleavage, cleavage of the side chain protecting group: adding 0.25ml triisopropylsilane, 0.25ml water, 0.25ml 3, 6-dioxa-1, 8-octanedithiol, 9.25ml trifluoroacetic acid, shaking at 40deg.C for 2 hr, filtering, and N 2 Blowing off trifluoroacetic acid, adding 30ml of absolute glacial diethyl ether, centrifuging at 3000rpm for 3min to obtain white precipitate, repeatedly washing with cold absolute diethyl ether for 3 times, and vacuum drying to obtain crude product.
(17) Purifying the crude product by reverse phase preparative liquid chromatography (RP-HPLC), collecting the target peak mobile phase solution, removing acetonitrile, and lyophilizing to obtain flocculent or powdery solid, i.e. BimBH3 analog peptide analog pure product, and performing structure confirmation by mass spectrometry and high performance liquid chromatography analysis.
Mass spectrum data and HPLC purity analysis data for the 25 BimBH3 mimetic peptide compounds described in the present invention are shown in table 1.
Table 1. Mass spectrometry data and HPLC purity analysis data for the simulated peptide analogues of bh 3.
Example 2: determination of protein tyrosine phospholipase 1B (PTP 1B) inhibitory Activity
According to the invention, MES buffer solution is adopted as a reaction system, human protein tyrosine phosphatase 1B (PTP 1B) is utilized, disodium p-nitrophenylphosphate (pNPP) is adopted as a specific substrate, a lead compound SM-6 is selected as a positive control, DMSO is adopted as a negative control, a screening model based on 96-hole microplates with enzyme reaction rate is established, and a PTP1B inhibitor is found through an enzymology method.
The specific implementation method comprises the following steps: using MES buffer (25 mM, pH 6.5), 10. Mu.L of pNPP (77 mM), 86. Mu.L of MES buffer, 4. Mu.L of compound (2 mM), 100. Mu.L of PTP1B solution (50 nM) were sequentially added to a 96-well plate, and the total volume of the reaction was 200. Mu.L. Each group of 3 parallels toDMSO is used as negative control, sodium orthovanadate (2 mM) is used as positive control, shake is carried out on a shaker for 1min at 25 ℃, and OD (optical density) of 1min and OD of 5min are respectively read on an enzyme label instrument 405 Value, calculate OD 405 The rate of change v (. DELTA.OD/min). The initial reaction rate of each well is linearly dependent, and the slope of the linear part of the kinetic curve determines the reaction rate of PTP1B, which is expressed as the rate of enzyme activity. For the data obtainedThe data from each group is shown using t-test analysis. The inhibition rate of the compound to PTP1B is calculated by the formula:
inhibition (%) = (v) DMSO -v Sample of )/v DMSO ×100％
Wherein v is DMSO 、v Sample of Initial average reaction rates of the negative control group and the test compound are shown, respectively. Statistical inhibition rate data statistics and processing were performed using GraphPad Prism software, inhibition dose response curves were fitted and IC was calculated 50 Values.
TABLE 2 inhibition of PTP1B Activity by test mimetic peptide analogs
* : compounds with a primary screening inhibition of less than 50% were not IC 50 Is measured.
As shown in Table 2, the BimBH3 mimetic peptide compounds of the present invention generally have excellent PTP1B inhibitory activity, particularly PTP1B inhibitory ICs of DFY-1, DFY-3, DFY-4, DFY-5, DFY-6, DFY-7 and DFY-19 50 The values are all lower than 400nmol/L, and the potential in vivo hypoglycemic activity is excellent.
Example 3: plasma stability evaluation of BimBH3 mimetic peptide compounds.
The high-activity PTP1B inhibitor mimic peptide is prepared into a certain concentration, then incubated with plasma, and samples at different time points are taken and quenched with a quencher (0.2% TFA/CH) 3 CN) quenching the reaction, and after workup, analyzing with analytical HPLC. Each time point phaseThe time origin residual compound (%) was determined by the relative peak area. Half-life was determined by plotting the amount of peptide remaining versus reaction time, fitting the data to an exponential decay model in GraphPad Prism software, to evaluate its stability.
TABLE 3.4 in vitro plasma half-life data for the high Activity PTP1B inhibitor BimBH3 mimetic peptides
As shown in Table 3, the 4 highly active PTP1B inhibitor BimBH3 mimetic peptide compounds of the present invention have half-lives exceeding 30 hours in rat and dog plasma ex vivo, and particularly the compound DFY-7 exhibits excellent stability in plasma ex vivo in both animals (t 1/2 ＝183.1h、t 1/2 =154.9h), can be kept in vivo and exert target inhibition activity continuously theoretically, and the action time can reach 1 week or even longer, which indicates that the BimBH3 mimetic peptide compound provided by the invention has overcome the problem of metabolic instability of polypeptide drugs. The compounds are suggested to have long-acting time in vivo.
Example 4: evaluation of in vivo long-acting hypoglycemic effect of high-activity PTP1B inhibitor BimBH3 mimic peptide compound
Evaluation of in vivo hypoglycemic effect: female C57BL/Ks db/db mice are selected, and are normally bred in animal houses with constant temperature of 24+/-2.0 ℃ and normal sunlight period at the age of 8-9 weeks. After 1 week of adaptive feeding, fasting blood glucose was measured, and mice with blood glucose levels exceeding 11mM were selected and randomly grouped, and a parallel control group (physiological saline), a positive control group (cable Ma Lutai, 0.5. Mu. Mol/kg) and a treatment group (BimBH 3 mimetic peptide compound, 0.5. Mu. Mol/kg) were set, respectively. 5 groups, 5 groups total, were administered once a week by subcutaneous injection (10 mL/10g mouse body weight) for a treatment period of 2 weeks. Blood glucose levels were measured using an ACCU-CHEK Performa glucometer (Roche Diabetes Care Gmbh Sandhofer Strasse 11668305Mannheim, germany) for tail blood sampling in mice and each group of type 2 diabetic model mice was monitored for changes in Fasting Blood Glucose (FBG) levels.
The results of the in vivo long-acting hypoglycemic activity of the 5 high-activity PTP1B inhibitor BimBH3 mimic peptide compounds are shown in figure 1. Each group was measured for pre-experimental fasting blood glucose levels on day 1, and each of the two groups was dosed once on day 2 and day 9, and as can be seen from the experimental results of fig. 1, the fasting blood glucose of the model mice of the parallel control group was continuously increased, which corresponds to the pathological characteristics of type 2 diabetes, whereas the fasting blood glucose levels of the mice of the treatment group at the blood glucose monitoring time points (day 9 and day 16) were significantly lower than those of the parallel control group, and the fasting blood glucose levels at the blood glucose monitoring time points (day 17, 19, 21 and 23) after stopping the treatment were still lower than those of the parallel control group, indicating that the BimBH3 mimetic peptide compound (DFY-1, DFY-3, DFY-6 and DFY-7) having excellent PTP1B inhibitory activity of the present invention exhibited a once-week long-acting therapeutic effect against type 2 diabetes, and had very high development prospects and application values.
The above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be apparent to one skilled in the art that modifications may be made to the technical solutions described in the foregoing embodiments, or equivalents may be substituted for some of the technical features thereof; such modifications and substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (10)
1. The invention discloses a novel long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B, and a preparation method and application thereof. The structural general formula of the BimBH3 mimic peptide analogue is shown in formula I:
BimBH3 mimetic peptide compounds of formula I.
Where n=14, raa is a different substituted natural or unnatural amino acid.
2. The novel PTP1B targeted long acting hypoglycemic BimBH3 mimetic peptide according to claim 1, wherein the substituted natural or unnatural amino acids at Raa position are respectively: gly, aib, ser, asp, lys, val, thr, phe, pro, hyp, leu, ile, trp, tyr, cys, met, glu, arg, D-Ala, beta-Ala, D-Ser, D-Leu, D-Pro, D-His and N-CH 3 -Ala。
3. The novel PTP1B targeted long acting hypoglycemic BimBH3 mimetic peptide according to claim 1, wherein the preparation method comprises the steps of:
(1) Placing CTC resin in a manual polypeptide solid-phase synthesizer at room temperature, and activating with dichloromethane and dimethylformamide;
(2) Adding a piperidine/dimethylformamide mixed solution to remove Fmoc protecting groups;
(3) Adding 2-3 times of N-Fmoc protected amino acid or fatty acid and 2.4-4.8 times of HOBt, DIC or HBTU, HOBt and DIEA in the molar amount of resin, and blowing air at 40 ℃ for reaction for 30-60 min;
(4) Repeating the steps (2) and (3) until the synthesis of the whole mimic peptide sequence is completed;
(5) Adding a lysate into the product obtained in the step (4), oscillating for 2 hours at 40 ℃, filtering, adding absolute ethyl glacial ether to separate out solid, and washing and vacuum drying to obtain a crude product of the analogue;
(6) And purifying the crude peptide analogue product by using reverse phase preparation liquid chromatography, collecting a target peak mobile phase solution, removing acetonitrile, and freeze-drying to obtain flocculent or powdery solid, thus obtaining the pure pam BH3 simulated peptide analogue product modified by palmitic acid.
4. The method according to claim 3, wherein the cleavage liquid in the step of producing comprises triisopropylsilane, 3, 6-dioxa-1, 8-octanedithiol, water and trifluoroacetic acid.
5. The method according to claim 3, wherein N is blown after filtration in the step (5) 2 Excess trifluoroacetic acid is removed.
6. A medicament or pharmaceutical composition comprising as active ingredient the novel, PTP 1B-targeted, long-acting hypoglycemic BimBH3 mimetic peptide according to any one of claims 1 to 5, wherein said medicament or pharmaceutical composition comprises any one of said novel, PTP 1B-targeted, long-acting hypoglycemic BimBH3 mimetic peptide and one or more pharmaceutically acceptable carriers or excipients.
7. Use of a BimBH3 mimetic peptide analogue according to any one of claims 1 to 5 in the manufacture of an inhibitor for inhibiting PTP1B activity.
8. Use of a BimBH3 mimetic peptide analogue according to any one of claims 1 to 5 in the manufacture of a medicament for the prophylaxis or treatment of a PTP1B targeted disease.
9. The use according to claim 8, characterized in that: such diseases include diabetes, cancer and Alzheimer's disease, with long-acting diabetes therapeutic drugs being preferred.
10. The use according to claim 8, characterized in that: the drug or the drug composition taking the BimBH3 mimetic peptide as an active ingredient is orally taken or injected.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311330112.9A CN117343145A (en) | 2023-10-13 | 2023-10-13 | Long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311330112.9A CN117343145A (en) | 2023-10-13 | 2023-10-13 | Long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117343145A true CN117343145A (en) | 2024-01-05 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311330112.9A Pending CN117343145A (en) | 2023-10-13 | 2023-10-13 | Long-acting hypoglycemic BimBH3 mimic peptide targeting PTP1B and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117343145A (en) |
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2023
- 2023-10-13 CN CN202311330112.9A patent/CN117343145A/en active Pending
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