CN109224075B

CN109224075B - Application of PTEN inhibitor in preparation of medicine for treating type 1 diabetes

Info

Publication number: CN109224075B
Application number: CN201810892206.8A
Authority: CN
Inventors: 翁勤洁; 杨波; 何俏军; 赵梦婷; 王佳颖; 郑佳焕; 王金成
Original assignee: Zhejiang University ZJU
Current assignee: Zhejiang University ZJU
Priority date: 2018-08-07
Filing date: 2018-08-07
Publication date: 2021-11-16
Anticipated expiration: 2038-08-07
Also published as: CN109224075A

Abstract

The invention provides an application of a PTEN inhibitor in preparing a medicament for treating type 1 diabetes, wherein small molecular compounds in the inhibitor are mainly peroxyorganic vanadium compounds, and are mainly used for treating cerebral/cardiac ischemia, repairing after nerve injury and the like at present. Animal experiment researches prove that the PTEN inhibitor does not influence the blood sugar and related sugar metabolism of normal mice and wild type 1 diabetes mice, only has obvious treatment effect on type 1 diabetes mice with deletion mutation of STAT3 in pancreatic beta cells, can effectively improve the blood sugar, increase the insulin secretion, reduce the beta cell apoptosis and reverse the diabetes. Therefore, the PTEN accumulation can be used as a specific marker protein for diagnosing STAT3 deletion mutant diabetes, so that the diagnosis and treatment of diabetes are more accurate and quicker. The PTEN inhibitor provides a precise treatment target and an effective new medicine for treating STAT3 deletion mutation diabetes.

Description

Application of PTEN inhibitor in preparation of medicine for treating type 1 diabetes

Technical Field

The invention belongs to the field of pharmacy, and relates to application of a PTEN inhibitor in preparation of a medicament for treating STAT3 deletion mutation type 1 diabetes.

Background

Diabetes is a chronic progressive disease with 1 high morbidity and mortality, needs lifelong medication along with various complications, and has a global morbidity of 8.5%. Islet beta cell death is the underlying cause of the development of type 1 diabetes and is an important factor in the development of type 2 diabetes. Type 1 diabetes is caused by a sudden decrease in the number of beta cells, a lack of insulin secretion, and abnormal regulation of blood glucose. Type 2 diabetes accounts for more than 90% of the total number, and islet beta cells of the body are dysfunctional in a hyperglycemic alicyclic environment, and cell masses are atrophied, so that insulin cannot be normally secreted. At present, the method for treating diabetes mellitus is generally used clinically, which is to regulate insulin secretion and mainly uses insulin, insulin secretion promoting medicaments (sulfonylureas, GLP-1 agonists and DPP-4 inhibitors), insulin sensitizers (biguanides, thiazolidinediones, SGTL-2 inhibitors) and other medicaments for treatment. All hypoglycemic drugs have various side effects, such as hypoglycemia, liver injury, heart injury, gastrointestinal reactions, anaphylaxis and the like. Therefore, personalized medicines or targets are always searched for, diabetes can be effectively treated, adverse effects are reduced, and accurate medical treatment is realized. No report on accurately treating the type 1 diabetes with STAT3 deletion mutation in islet beta cells by inhibiting PTEN exists in clinic and clinical tests, so that the application and mechanism of the PTEN inhibitor in the type 1 diabetes with STAT3 deletion mutation are clear, and the method has very practical significance for treating individualized diabetes.

PTEN is a protein and lipid phosphatase that is currently reported to regulate pancreatic islet beta cell proliferation, carbohydrate metabolism, insulin sensitivity, and the like. The PTEN small molecular compound inhibitor is a peroxy organic vanadium compound, and the research is mainly focused on the treatment effect on the ischemia, such as experimental stroke, myocardial ischemia and cerebral ischemia, and is beneficial to nerve repair of spinal nerves, optic nerves and the like and lung epithelial cell repair. However, PTEN is not targeted so far, and reports related to the treatment of diabetes by using PTEN inhibitors are available.

Disclosure of Invention

The invention provides an application of a PTEN inhibitor in preparation of a medicament for treating type 1 diabetes, wherein the type 1 diabetes is type 1 diabetes caused by STAT3 deletion mutation.

The PTEN inhibitors include: small molecule compounds that inhibit the function of the PTEN protein, double-stranded ribonucleic acids that inhibit the expression of the PTEN gene by RNA interference, or antibody proteins for inhibiting the activity of the PTEN protein. The PTEN protein can be used as a new target point for STAT3 deficiency diabetes treatment, and promotes personalized diabetes treatment. The PTEN gene maps to chromosome 10q23, spans a genomic region of approximately 218k, contains 9 exons and 8 introns, and has an mRNA length of 5500 bp.

The small molecular compound for inhibiting the function of the PTEN protein comprises the following peroxy compoundsAn organic vanadium compound: bpv (phen), diperoxy (1, 10-phenanthroline) potassium vanadyl, molecular weight 404.29, molecular formula C₁₂H₈KN₂O₅V.3H₂O; bpv (pic), dipotassium diperoxy (pyridinone) vanadates, molecular weight 367.20, formula C₆H₄KNO₇V.2H₂O; bpv (bipy), potassium diperoxy (bipyridine) vanadyl, molecular weight 326.22, molecular formula C₁₀H₈KN₂O₅V; bpv (HOpic), Potassium bis (5-hydroxypyridine-2-carboxy) oxovanadate, molecular weight 347.24, molecular formula C₆H₄K₂NO₈V; VO-Ohpic, hydroxy (oxo) vanadium 3-hydroxypyridine-2-carboxylic acid, molecular weight 415.20, molecular formula C₁₂H₉N₂O₈V.3H₂O.H are provided. Also included are other reported or potential PTEN inhibitors such as SF1670, N- (9, 10-dioxo-9, 10-dihydrophenanthrene-2-) pivalamide, molecular weight 307.34, formula C₁₉H₁₇NO₃. In an in vivo animal model, the mouse pharmaceutical dose of the PTEN inhibitor is 1mg/kg-20 mg/kg.

The medicament is prepared from a PTEN inhibitor and pharmaceutically acceptable auxiliary materials, and the medicament dosage form is a gel, a soft capsule, an oral preparation, an injection, a freeze-dried powder injection or a large infusion.

In the present invention, a series of methods known in the art can be used to prepare antibody proteins specific for PTEN proteins. For example, purified human PTEN protein or an antigenic fragment thereof is injected into an animal to produce polyclonal antibodies. Similarly, cells expressing the human PTEN protein or an antigenic fragment thereof can be used to immunize an animal to produce antibodies. The antibodies produced according to the invention may also be monoclonal antibodies, which may be produced using hybridoma technology. The antibodies of the invention include, inter alia, antibodies that inhibit the function of PTEN and can be produced by immunizing a fragment or domain of the human PTEN protein, and the human PTEN protein product and fragments thereof can be produced recombinantly or synthesized using a polypeptide synthesizer. Antibodies that bind to unmodified forms of PTEN protein can be obtained by immunizing an animal with a gene product produced in a prokaryotic cell. Antibodies that bind to post-translationally modified forms, such as glycosylated or phosphorylated PTEN proteins or polypeptides, can be obtained by immunizing an animal with a gene product produced in a eukaryotic cell.

The experiment proves that the PTEN small molecular compound inhibitor has obvious therapeutic effect on the type 1 diabetes mouse of beta-STAT 3KO induced by Streptozotocin (STZ). The PTEN small molecular compound inhibitor bpv (phen) reverses the blood sugar of a beta-STAT 3KO mouse, improves oral glucose tolerance, protects beta cell function, prevents beta cell apoptosis, and has no treatment effect on a non-STAT 3-deleted type 1 diabetes mouse. At present, no better medicine plays a role in accurate treatment by regulating PTEN clinically, and related research and invention are not found. Therefore, the PTEN inhibitor can treat type 1 diabetes under the STAT3 deficiency condition, and provides a new treatment target and a new effective drug for the precise treatment of diabetes.

Drawings

FIG. 1bpv-

phen

1, 2, 5, 10, 15, 20 on the blood glucose lowering effect of STZ-induced β -STAT3KO diabetic mice.

FIG. 2bpv-

phen

1, 2, 5, 10, 15, 20 action on oral glucose tolerance improvement in STZ-induced β -STAT3KO diabetic mice.

FIG. 3bpv-phen 2mg/kg on STZ-induced increase in insulin secretion in β -STAT3KO diabetic mice.

FIG. 4bpv-phen 2mg/kg insulin bolus protection in STZ-induced β -STAT3KO diabetic mice.

FIG. 5bpv-phen 2mg/kg insulin protective effect on STZ-induced β -STAT3KO diabetic mice.

FIG. 6bpv-phen 2mg/kg had no effect on STZ-induced blood glucose in wild type diabetic mice.

FIG. 7bpv-phen 2mg/kg had no effect on STZ-induced oral glucose tolerance in wild type diabetic mice.

Detailed Description

The invention is further explained by the accompanying drawings and examples.

Example 1: selecting 8-10 week old male mice with genotypes of beta-STAT 3KO and C57BL/6, continuously molding STZ for three days at a small dose (40mg/kg), setting a corresponding normal control group, starving for 12 hours before administration and molding, and constructing a type 1 diabetes model. And detecting random blood sugar on the third day after molding, and detecting for 1 time every 1 day. The model groups of beta-STAT 3KO and C57BL/6 are divided into the administration group and the model group of the PTEN small molecular compound inhibitor bpv-

phen

1, 2, 5, 10, 15 and 20mg/kg according to random blood sugar. Bpv-phen 2mg/kg is injected into the abdominal cavity 10 days after the model is made, and the medicine is killed 28 days after the medicine is administered. As shown in figures 1-5, the bpv-phen administration group obviously improves blood sugar, oral glucose tolerance, insulin secretion, islet structure and mass of STAT3 deletion mutant diabetic mice.

Example 2: selecting 8-10 weeks old C57BL/6 male mice, STZ150mg/kg single molding, setting corresponding normal control group, starving for 12 hours before administration molding, and constructing type 1 diabetes model. As shown in FIGS. 6-7, the bpv-phen administration group had no effect on blood glucose and oral glucose tolerance in the normal diabetic mice.

The experimental results are as follows:

referring to fig. 1, bpv-

phen

1, 2, 5, 10, 15 and 20mg/kg of beta-STAT 3KO model mice can obviously reduce the blood sugar of the diabetic mice.

Referring to fig. 2, bpv-

phen

1, 2, 5, 10, 15 and 20mg/kg of beta-STAT 3KO model mice can obviously improve the oral glucose tolerance of the diabetic mice.

Referring to fig. 3, bpv-phen 2mg/kg of beta-STAT 3KO model mice can obviously promote insulin secretion of the diabetic mice.

Referring to fig. 4, bpv-phen 2mg/kg of beta-STAT 3KO model mice can obviously protect the islet mass of the diabetic mice and prevent beta cell death.

Referring to fig. 5, bpv-phen 2mg/kg of beta-STAT 3KO model mice can obviously improve the islet structure of the diabetic mice.

Referring to FIG. 6, bpv-phen 2mg/kg was administered to WT mice, and had no effect on blood glucose in diabetic mice.

Referring to FIG. 7, bpv-phen 2mg/kg was administered to WT mice, and there was no effect on oral glucose tolerance in diabetic mice.

The invention achieves the aim of developing accurate therapeutic drugs by treating STAT3 deletion mutant type 1 diabetes through a PTEN inhibitor. At the animal level, the PTEN small molecule inhibitor bpv (phen) is effective in reversing STAT3 deletion mutant type 1 diabetes.

The invention has the significance of providing the micromolecule compound for potentially treating STAT3 deletion mutation type 1 diabetes, thereby promoting the accurate medical development of diabetes, realizing personalized treatment and lightening the adverse reaction of clinical medicines.

Claims

1. An application of a PTEN inhibitor in preparation of a drug for treating type 1 diabetes is characterized in that the PTEN inhibitor is applied in preparation of a drug for treating type 1 diabetes under a STAT3 deficiency condition, the PTEN inhibitor is a peroxy organic vanadium compound, and the peroxy organic vanadium compound is selected from diperoxy (1, 10-phenanthroline) potassium vanadyl acid.

2. The use as claimed in claim 1 wherein the pharmaceutically acceptable dose of the PTEN inhibitor is from 1mg/kg to 20mg/kg in an in vivo animal model.

3. The use of claim 1, wherein the medicament is prepared from a PTEN inhibitor and pharmaceutically acceptable excipients, and the medicament is in the form of a gel, a soft capsule, an injection or a lyophilized powder for injection.