阅读说明：本技术 一种磺胺类化合物及其在制备治疗糖尿病和并发症药物中的应用 (Sulfonamide compound and application thereof in preparing medicine for treating diabetes and complications ) 是由 陈新平 李纯 吴桐雨 于 2021-09-06 设计创作，主要内容包括：本发明涉及医药技术领域,具体涉及一种磺胺类化合物及其在制备治疗糖尿病和并发症药物中的应用。本发明所述的磺胺类化合物的结构式如式(Ⅰ)所示,实验结果显示其可以改善2型糖尿病小鼠的糖代谢,提高胰岛素的敏感性,改善胰岛素抵抗,将其用于治疗2型糖尿病小鼠,其降血糖的疗效是二甲双胍的200倍或更高；降低2型糖尿病小鼠的尿蛋白、尿肌酐和尿素氮的含量,进而说明磺胺类化合物可以保护糖尿病小鼠的肾功能,治疗糖尿病引起的肾病；降低2型糖尿病小鼠的甘油三酯和总胆固醇的含量,调节血脂水平,改善糖尿病小鼠的脂代谢,说明磺胺类化合物可以治疗糖尿病心血管类并发症,可以在临床上推广应用。(The invention relates to the technical field of medicines, in particular to a sulfonamide compound and application thereof in preparing medicines for treating diabetes and complications. The sulfonamide compound has a structural formula shown in a formula (I), and experimental results show that the sulfonamide compound can improve the glucose metabolism of type 2 diabetes mice, improve the sensitivity of insulin and improve insulin resistance, and when the sulfonamide compound is used for treating type 2 diabetes mice, the curative effect of reducing blood sugar is 200 times or more that of metformin; reducing the contents of urine protein, urine creatinine and urea nitrogen in a type 2 diabetic mouse, thereby indicating that the sulfonamide compound can protect the renal function of the diabetic mouse and treat nephropathy caused by diabetes; the contents of triglyceride and total cholesterol of a type 2 diabetic mouse are reduced, the blood fat level is regulated, the lipid metabolism of the diabetic mouse is improved, and the sulfonamide compound can treat cardiovascular complications of diabetes and can be clinically popularized and applied.)

1. The application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the medicament for treating the diabetes mellitus;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

2. The use of claim 1, wherein R is₄，R₅，R₆，R₇Are each selected from hydrogen atoms.

3. The use of claim 2, wherein R is₁、R₂Are each selected from hydrogen atoms.

4. The use of claim 3, wherein the diabetes is type 2 diabetes or early stage type 1 diabetes.

5. The use of a sulfonamide compound of formula (i) or a tautomer, mesomer, racemate, enantiomer, diastereomer or mixture thereof, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of diabetic complications;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano,Carboxy, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

6. The use of claim 5, wherein the diabetic complication comprises one or more of gestational diabetes, microangiopathy, skin ulcer, cardiovascular disease and periodontitis.

7. The use of claim 6, wherein the microvascular disease is nephropathy or retinopathy.

8. The use according to any one of claims 1 to 7, wherein the medicament is administered by the following route: sublingual, inhalation, oral administration or injection.

9. The application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the health care product with the function of assisting in reducing blood sugar;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy or cycloalkylHalogen, amino, hydroxyl, carboxyl, cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

10. The application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture thereof, or the pharmaceutically acceptable salt thereof in preparing the health care product with the function of assisting in reducing blood fat;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

Technical Field

The invention relates to the technical field of medicines, in particular to a sulfonamide compound and application thereof in preparing medicines for treating diabetes and complications.

Background

Diabetes is a metabolic disease characterized by hyperglycemia. Hyperglycemia is caused by a defect in insulin secretion or an impaired biological action, or both. Diabetes is a major disease that endangers human health and can be divided into type 1 diabetes and type 2 diabetes. The international diabetes association estimates that about 3.87 million people are currently diagnosed with diabetes, with type 2 diabetes accounting for over 90%, and the estimated number of people will reach 5.95 million by 2035 years. Type 2 diabetes is a type of diabetes mellitus in which insulin is relatively hyposecreted, and/or insulin resistance is the main cause. The relative hyposecretion of insulin refers to that the carbohydrate ingested by the type 2 diabetes mellitus patient exceeds the maximum degree regulated by the body to secrete insulin, so that the continuous high glucose in blood is caused; insulin resistance refers to the rise in blood glucose caused by a decrease in the sensitivity of insulin secreted by the patient's islet beta cells in their effector cells (e.g., muscle cells, adipocytes, liver cells, etc.). Currently, treatment regimens for diabetes include drug therapy, dietary therapy, exercise therapy, and general therapy. Available drugs mainly comprise biguanides, sulfonylureas, Thiazolidinediones (TZD), meglitinide, dipeptidyl peptidase 4(DPP-4) inhibitors, sodium-glucose cotransporter (SGLT2) inhibitors, alpha-glucosidase inhibitors and insulin preparations, but the current treatment status of diabetes is not ideal, the control rate of diabetes in China is 39.7%, the treatment rate is 25.8%, and the 2010 sample epidemiological research shows that the standard reaching rate is only 16.8% and patients with unqualified blood sugar control reach 83.2% by taking glycosylated hemoglobin < 6.5% as the control standard.

Meanwhile, a series of complications such as diabetic nephropathy, diabetic skin ulcer, retinopathy, cardiovascular diseases and the like can be generated after the long-term development of diabetes, so that a large economic burden is brought to patients and the society, the health of human is seriously harmed, and the living quality of the patients is influenced. The diabetic nephropathy, the diabetic cardiovascular disease, the diabetic retinopathy and the diabetic skin ulcer are the most serious complications of diabetes, and no ideal medicine exists for clinically treating the diabetic complications.

The sulfanilamide type drug is an artificially synthesized antibacterial drug, is used for nearly 50 years in clinic, and has the advantages of wide antibacterial spectrum, stable property, simple and convenient use, no grain consumption in production and the like. Particularly, after the discovery of Trimethoprim (TMP), an antibacterial synergist in 1969, the combined application of trimethoprim and sulfanilamide can enhance the antibacterial effect and enlarge the treatment range, so that the sulfanilamide is an important chemotherapeutic drug although a large number of antibiotics are available. Sulfonamides have inhibitory effects on many gram-positive and some gram-negative bacteria, Nocardia, Chlamydia and certain protozoa (e.g. Plasmodium and amebiasis). In view of the new use of sulfonamides, researchers have also conducted many studies, for example, a patent (CN201010581743.4) discloses the use of sulfadiazine sodium in the preparation of a medicament for preventing or treating silkworm sepsis, and a patent (CN201410477735.3) discloses the use of a sulfonamide composition in the preparation of a medicament for treating foot odor; the patent (cn201410477958.x) discloses the use of pharmaceutical compositions of sulfonamides for the preparation of a medicament for the treatment of bromhidrosis. However, no researchers have studied the application of sulfonamides to treat diabetes and its complications, and no literature has disclosed the use of sulfonamides to treat diabetes and its complications.

The inventor unexpectedly discovers in the research process that the sulfonamide medicaments have the curative effect of treating the diabetes and the curative effect of treating the diabetic complications.

Disclosure of Invention

Aiming at the technical problems, the invention provides a sulfonamide compound and application thereof in preparing medicines for treating diabetes and complications. The specific technical scheme is as follows:

the first purpose of the invention is to provide the application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the medicine for treating diabetes;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy or cyclicAlkyl, halogen, amino, hydroxyl, carboxyl, cyano substituted;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

Preferably, R is₄，R₅，R₆，R₇Are each selected from hydrogen atoms.

Preferably, R is₁、R₂Are each selected from hydrogen atoms.

Preferably, the compounds of formula (I) include, but are not limited to

Preferably, the diabetes is type 2 diabetes or early stage type 1 diabetes.

The second purpose of the invention is to provide the application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the medicine for treating the diabetic complication;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

Preferably, the diabetic complication comprises one or more of gestational diabetes, microangiopathy, skin ulcer, cardiovascular disease and periodontitis.

Preferably, the microvascular disorder is renal disorder and retinopathy.

Preferably, the medicament is administered by the following route: sublingual, inhalation, oral administration or injection.

The third purpose of the invention is to provide the application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the health care product with the auxiliary hypoglycemic function;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

The fourth purpose of the invention is to provide the application of the sulfonamide compound shown in the formula (I) or the tautomer, the mesomer, the racemate, the enantiomer, the diastereomer or the mixture form thereof, or the pharmaceutically acceptable salt thereof in preparing the health care product with the function of assisting in reducing blood fat;

R₁selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₂selected from hydrogen atoms, C_1-6Alkyl radical, C_1-8Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring and the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₁and R₂N atom which may be bonded thereto forms a five-or six-membered heterocyclic ring;

R₃selected from hydrogen atoms, C_1-6Alkyl radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, benzene ring, five-membered or six-membered aromatic heterocycle can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano;

R₄，R₅，R₆，R₇are respectively selected from hydrogen atom, amino, substituted amino, hydroxyl, cyano, carboxyl and C_1-6Alkyl radical, C_1-6Alkoxy radical, C_3-6Cycloalkyl, a benzene ring, a five-or six-membered aromatic heterocycle; wherein, the benzene ring, the five-membered or six-membered aromatic heterocyclic ring can be substituted by alkyl, alkoxy, cycloalkyl, halogen, amino, hydroxyl, carboxyl and cyano.

The invention has the beneficial effects that: the invention provides a new application of a sulfanilamide compound, the sulfathiazole can improve the glucose metabolism of a type 2 diabetes mouse, improve the sensitivity of insulin, improve insulin resistance, and obviously reduce the blood sugar of the type 2 diabetes mouse, and the blood sugar reducing effect of the sulfanilamide compound is 200 times or more that of metformin; meanwhile, the sulfanilamide compound sulfathiazole can reduce the contents of urine protein, urine creatinine and urea nitrogen of a type 2 diabetes mouse, which indicates that the sulfanilamide compound sulfathiazole can protect the renal function of the diabetes mouse; the sulfanilamide compound sulfathiazole can reduce the content of triglyceride and total cholesterol of a type 2 diabetes mouse, regulate the blood fat level and improve the lipid metabolism of the diabetes mouse. The sulfa compound sulfadiazine can obviously reduce the blood sugar of a type 2 diabetic mouse and has a therapeutic effect on the type 2 diabetic mouse; the sulfanilamide compound sulfamethazine can obviously reduce the blood sugar of a type 2 diabetes mouse, and has a treatment effect on the type 2 diabetes mouse. The sulfonamide compound can be used for treating cardiovascular complications of diabetes and can be clinically popularized and applied.

Drawings

FIG. 1 weight changes in mice of each group

FIG. 2 blood glucose changes in groups of mice

FIG. 3 comparison of urine proteins in mice of each group

FIG. 4 comparison of urine creatinine in mice of each group

FIG. 5 comparison of urea nitrogen content in groups of mice

FIG. 6 shows the results of 0-120min blood glucose changes in mice of each group

FIG. 7 area under 0-120min blood glucose Change Curve of each group of mice

FIG. 8 results of blood glucose changes of each group of mice at 0-120min

FIG. 9 area under 0-120min blood glucose Change Curve of each group of mice

FIG. 10 comparison of triglyceride levels in mice of each group

FIG. 11 comparison of low-density lipoprotein cholesterol levels in mice of each group

FIG. 12 comparison of Total Cholesterol levels in groups of mice

FIG. 13 weight changes in mice of each group

FIG. 14 blood glucose changes in groups of mice

FIG. 15 changes in body weight of mice in each group

FIG. 16 blood glucose changes in groups of mice

Detailed Description

The following detailed description of the present invention will be provided in connection with specific embodiments thereof, but it should be understood that the present invention is not limited to the following examples.

Metformin hydrochloride, its chemical name is: 1.1-dimethylbiguanide hydrochloride for type 2 diabetes patients with inadequate simple dietary control, especially obesity and hyperinsulinemia patients, and has effects of reducing blood glucose, and reducing body weight and hyperinsulinemia. Can be used for treating patients with poor curative effect of some sulfonylureas, such as sulfonylureas, small intestine glycosidase inhibitor or thiazolidinedione hypoglycemic agent, which has better effect than single use. Can also be used for patients with insulin therapy to reduce insulin dosage. However, the metformin hydrochloride cannot cure type 2 diabetes, and has the advantages of large dosage, long medication period, large side effect and no treatment effect on diabetic complications.

Streptozotocin, also called streptozocin (streptozocin), has the chemical name of 2-deoxy-2- [ [ (methyl nitrosoamino) carbonyl ] -amino ] -D-glucopyranose, the molecular formula of C8H15N3O7 and the molecular weight of 265.22100, is light yellow crystalline powder which is easy to dissolve in water, but the aqueous solution of the streptozotocin is extremely unstable at room temperature and can be decomposed into gas after half an hour to be volatilized, so the streptozocin needs to be prepared at present. Soluble in lower alcohols and ketones, insoluble in polar organic solvents. The streptozotocin can damage beta cells of pancreatic islets of animals and reduce insulin secretion, so the streptozotocin can be used for inducing a diabetes animal model, generally, 1-type diabetes can be induced by single injection of a large dose (150mg/kg/day), and 2-type diabetes can be induced by multiple injections (3-5 days) of a small dose (40-60 mg/kg/day) in combination with high-fat feed feeding.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited herein are incorporated by reference in their entirety unless otherwise indicated. When a trade name appears herein, it is intended to refer to its corresponding commodity or its active ingredient.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the subject matter herein. In this application, it must be noted that, unless the context clearly dictates otherwise, as used in this specification and the claims, the singular forms "a," "an," and "the" include plural referents. It should also be noted that the use of "or", "or" means "and/or" unless stated otherwise. Furthermore, the term "comprising" as well as other forms, such as "includes," "including," and "containing," are used without limitation.

In the following examples of the present invention, the following terms full Chinese, full English or short may be used, but whether full Chinese, full English or short, refer to the same compound or drug or reagent. The method comprises the following specific steps:

chinese and English comparison abbreviation word list

Unless a specific definition is set forth, the nomenclature and laboratory procedures and techniques related to analytical chemistry, synthetic organic chemistry, and chemistry such as medical and pharmaceutical chemistry are known to those skilled in the art. Standard techniques can be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation, drug delivery and treatment of patients. Standard techniques can be used for recombinant DNA, oligonucleotide synthesis, and tissue culture and transformation (e.g., electroporation, lipid infection). For example, the reaction and purification techniques can be carried out using a kit with instructions provided by the manufacturer, or according to methods known in the art, or according to the methods described herein. In general, the foregoing techniques and procedures may be practiced by conventional methods well known in the art and described in various general or more specific documents that are cited and discussed in this disclosure.

Reagents used in the invention

Material for use in the present invention

The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not. For example, ethyl is "optionally" substituted with halo, meaning that ethyl may be unsubstituted (-CH)₂CH₃) Monosubstituted (e.g. -CH)₂CH₂F) Polysubstituted (e.g. -CHFCH)₂F、-CH₂CHF₂Etc.) or fully substituted (-CF)₂CF₃). It will be appreciated by those skilled in the art that any group containing one or more substituents will not incorporate any substitution or substitution pattern which is sterically impossible and/or cannot be synthesized.

The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced with a substituent, so long as the valence of the particular atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., ═ O), meaning that two hydrogen atoms are substituted, oxo does not occur on the aryl.

When any variable (e.g., R) occurs more than one time in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2R, the group may optionally be substituted with up to two R, and there are separate options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

The term "alkyl" refers to an optionally substituted straight or optionally substituted branched chain saturated aliphatic hydrocarbon group attached to the rest of the molecule by a single bond. The "alkyl" groups herein may have from 1 to about 8 carbon atoms, for example from 1 to 6 carbon atoms, or from 1 to 4 carbon atoms or from 1 to 3 carbon atoms. Examples of "alkyl" herein include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-2-propyl, 2-methyl-1-butyl, 3-methyl-l-butyl, 2-methyl-3-butyl, 2-dimethyl-1-propyl, 2-methyl-1-pentyl, 3-methyl-1-pentyl, 4-methyl-l-pentyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 2-dimethyl-l-butyl, 3-dimethyl-1-butyl, n-propyl, isopropyl, 2-methyl-l-propyl, 2-methyl-1-pentyl, 3-methyl-2-pentyl, 2-dimethyl-l-butyl, 3-dimethyl-1-butyl, 2-methyl-2-pentyl, and the like, 2-ethyl-1-butyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, hexyl and the like, and longer alkyl groups such as heptyl and octyl and the like. When a group as defined herein, such as "alkyl" comes within the numerical range, for example, "C1-8 alkyl" refers to an alkyl group that can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and for further example, "C1-4 alkyl" refers to an alkyl group that can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms. Alkyl herein also includes the case where no numerical range is specified.

The term "alkenyl" refers to an optionally substituted straight or optionally substituted branched monovalent hydrocarbon radical having at least one C ═ C double bond. The alkenyl group has, but is not limited to, 2 to 8 carbon atoms, such as 2 to 6 carbon atoms, 2 to 4 carbon atoms. The double bond in these groups may be in either the cis or trans conformation and should be understood to encompass both isomers. Examples of alkenyl groups include, but are not limited to, ethenyl (CH ═ CH)₂) 1-propenyl (CH)₂CH＝CH₂) Isopropenyl (C (CH)₃)＝CH₂) Butenyl, 1, 3-butadienyl and the like. When a numerical range is present for alkenyl as defined herein, e.g. "C_2-8The "alkenyl group" means an alkenyl group which may be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and the alkenyl group herein also covers the case where a numerical range is not specified.

The term "alkynyl" isRefers to an optionally substituted, straight or branched chain, monovalent hydrocarbon radical having at least one C.ident.C triple bond. The alkynyl group has, but is not limited to, 2 to 8 carbon atoms, such as 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Examples herein include, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and 1, 3-butadiynyl, and the like. When a numerical range occurs for alkynyl as defined herein, for example "C_2-8Alkynyl "refers to an alkynyl group that can be composed of 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, and alkynyl groups herein also encompass instances where a range of numbers is not specified.

The term "cycloalkyl" refers to a non-aromatic carbon-containing ring, including saturated carbocycles (e.g., cycloalkyl) or unsaturated carbocycles (e.g., cycloalkenyl). Carbocycle includes monocyclic (having one ring), and may be, for example, monocyclic cycloalkyl; a bicyclic carbocycle (having two rings), for example, may be a bicyclic cycloalkyl; carbocyclic (having more than two rings). The rings may be bridged or spiro. Carbocycle (e.g., cycloalkyl or cycloalkenyl) can have 3 to 8 carbon atoms, for example, 3 to about 6 ring-forming carbon atoms or 3 to about 5 ring-forming carbon atoms.

The term "aryl" refers to optionally substituted aromatic hydrocarbon groups having from about 6 to 20, such as 6 to 12 or 6 to 10 ring-forming carbon atoms, which may be monocyclic aryl, bicyclic aryl or higher ring aryl. The bicyclic aryl or higher ring aryl may be a monocyclic aryl fused to other independent rings such as alicyclic, heterocyclic, aromatic ring, aromatic heterocyclic. Non-limiting examples of monocyclic aryl groups include monocyclic aryl groups of 6 to about 12, 6 to about 10, or 6 to about 8 ring-forming carbon atoms, such as phenyl; bicyclic aryl is for example naphthyl; polycyclic aryl radicals are, for example, phenanthryl, anthracyl, azulenyl.

The term "heteroaryl" refers to optionally substituted heteroaryl groups containing from about 5 to about 20, such as 5 to 12 or 5 to 10, backbone ring-forming atoms, wherein at least one (e.g., 1-4, 1-3, 1-2) ring-forming atoms is a heteroatom independently selected from the group consisting of heteroatoms of oxygen, nitrogen, sulfur, phosphorus, silicon, selenium and tin, but is not limited thereto. Heteroaryl includes monocyclic heteroaryl (having one ring), bicyclic heteroaryl (having two rings), or polycyclic heteroaryl (having more than two rings). In embodiments where two or more heteroatoms are present in the ring, the two or more heteroatoms may be the same as each other, or some or all of the two or more heteroatoms may be different from each other. The bicyclic heteroaryl or higher ring heteroaryl may be a monocyclic heteroaryl fused with other independent rings such as alicyclic ring, heterocyclic ring, aromatic heterocyclic ring (which may be collectively referred to as fused ring heteroaryl). Non-limiting examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, quinolinyl, isoquinolinyl, tetrazolyl, triazolyl, triazinyl, benzofuranyl, benzothienyl, indolyl, isoindolyl, and the like.

The term "heterocyclyl" refers to a non-aromatic heterocyclic ring, which includes saturated or unsaturated heterocyclic rings (containing unsaturation), does not have a completely conjugated pi-electron system, and can be classified as a monocyclic, fused polycyclic, bridged or spiro ring system without aromaticity. Wherein one or more (e.g., 1-4, 1-3, 1-2) ring-forming atoms are heteroatoms, such as oxygen, nitrogen or sulfur atoms. Heterocycles can include mono-heterocycles (having one ring) or bis-heterocycles (having two bridged rings) or polyheterocycles (having more than two bridged rings); spiro rings are also included. A heterocyclyl group can have 3 to about 20 ring-forming atoms, such as 3 to about 10, 3 to about 8, 4 to 7, 5 to about 8, or 5 to about 6 ring-forming atoms. Non-limiting examples of heterocyclyl groups include oxiranyl, thietanyl, aziridinyl, azetidinyl, oxetanyl, thietanyl, tetrahydrofuryl, pyrrolidinyl, oxazolidinyl, tetrahydropyrazolyl, pyrrolinyl, dihydrofuranyl, dihydrothienyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, piperazinyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyranyl, dihydrothiopyranyl, azepanyl, oxepanyl, thiepanyl, thiepinyl, oxaazabicyclo [2.2.1] heptyl, and azaspiro [3.3] heptyl groups and the like.

The term "halo" or "halogen" refers to an optionally substituted group (e.g., alkyl, alkenyl, alkynyl, alkoxy, etc.) wherein at least one hydrogen atom is replaced with a halogen (e.g., fluorine, chlorine, bromine, iodine, or combinations thereof). In some embodiments, two or more hydrogens are replaced with the same halogen as each other (e.g., difluoromethyl, trifluoromethyl); in other embodiments two or more hydrogens are replaced with halogens that are not exactly the same as each other (e.g., 1-chloro-1-fluoro-1-iodoethyl).

The term "alkoxy" refers to an alkyl ether group (O-alkyl), non-limiting examples of which include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy and the like.

The term "alkanoyl" refers to a group wherein an alkyl group is attached to the group-CO-, non-limiting examples of which include formyl, acetyl, propionyl, butyryl and the like. For example, the term "C_1-6Alkylacyl "means C_1-6Alkyl groups are linked to-CO-to form groups. As another example, the term "C_1-4Alkylacyl "means C_1-4Alkyl groups are linked to-CO-to form groups.

The term "alkylsulfonyl" refers to alkyl and-SO₂-linked to form a group, non-limiting examples of which include methylsulfonyl, ethylsulfonyl, propylsulfonyl, butylsulfonyl and the like. For example, the term "C_1-6Alkylsulfonyl "means C_1-6Alkyl and-SO₂-linked to form a group. As another example, the term "C_1-4Alkylsulfonyl "means C_1-4Alkyl and-SO₂-linked to form a group.

The term "amino" refers to the group-NH₂Group, -NH (C)_1～6Alkyl) group or-N (C)_1～6Alkyl radical)₂A group. Specific examples of amino groups include, but are not limited to, -NH₂、-NHCH₃、-N(CH₃)₂、-NHC₂H₅、-N(C₂H₅)₂、-N(C₃H₇)₂、-N(CH₃)C₂H₅And the like.

The term "member" refers to the number of backbone atoms that make up a ring. For example, pyridine is a six-membered ring and pyrrole is a five-membered ring.

The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutical composition" refers to a biologically active compound optionally mixed with at least one pharmaceutically acceptable chemical ingredient or agent, i.e., a "carrier," which aids in the introduction of the compound into cells or tissues, including but not limited to stabilizers, diluents, suspending agents, thickening agents, and/or excipients.

The term "pharmaceutically acceptable salt" refers to salts that retain the biological potency of the free acid and free base of the specified compound, and that are biologically or otherwise not adversely affected. As the salt in the present invention, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like can be mentioned unless otherwise specified. Non-limiting examples of metal salts include, but are not limited to, salts of alkali metals, such as sodium, potassium, and the like; salts of alkaline earth metals such as calcium, magnesium, barium, and the like; aluminum salts, and the like. Non-limiting examples of salts with organic bases include, but are not limited to, salts with trimethylamine, triethylamine, pyridine, picoline, 2, 6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, and the like. Non-limiting examples of salts with inorganic acids include, but are not limited to, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, and the like. Non-limiting examples of salts with organic acids include, but are not limited to, salts with formic acid, acetic acid, trifluoroacetic acid, fumaric acid, oxalic acid, malic acid, maleic acid, tartaric acid, citric acid, succinic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, and the like. Non-limiting examples of salts with basic amino acids include, but are not limited to, salts with arginine, lysine, ornithine, and the like. Non-limiting examples of salts with acidic amino acids include, but are not limited to, salts with aspartic acid, glutamic acid, and the like.

Pharmaceutically acceptable salts can be synthesized from the parent compound, which contains an acid or base, by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

The term "treating" and other similar synonyms include alleviating, alleviating or ameliorating a symptom of a disease or disorder, preventing other symptoms, ameliorating or preventing the underlying metabolic cause of the symptom, inhibiting the disease or disorder, e.g., arresting the development of the disease or disorder, alleviating the disease or disorder, ameliorating the disease or disorder, alleviating a symptom caused by the disease or disorder, or halting a symptom of the disease or disorder, and further, the term can be included for prophylactic purposes. The term also includes obtaining a therapeutic effect and/or a prophylactic effect. The therapeutic effect refers to curing or ameliorating the underlying disease being treated. In addition, a cure or amelioration of one or more physiological symptoms associated with the underlying disease is also a therapeutic effect, e.g., an improvement in the condition of the patient is observed, although the patient may still be affected by the underlying disease. For prophylactic effect, the composition or compound can be administered to a patient at risk of developing a particular disease, or to a patient presenting with one or more physiological symptoms of the disease, even if a diagnosis of the disease has not yet been made.

EXAMPLE I Effect of Sulfathiazole in the treatment of diabetes mellitus and complications thereof in mice

1. Animal feeding

Experimental animals: 8 week-old SPF grade C57BL/6N male mice, weighing 18-22 g, never used any drug before the experiment, purchased from Lanzhou veterinary research institute of Chinese academy of agricultural sciences. The experimental animals are adaptively raised for one week in an environment with 24-26 ℃ and 12h/12h alternating day and night rules, and then fed with diet and free drinking water, and then the grouping experiment is carried out.

2. Drugs and reagents:

the medicines used in the invention are as follows: sulfathiazole (Sulfathiazole); metformin hydrochloride (Metformin hydrochloride, Metformin); streptozotocin (streptazocin, STZ).

Preparation of a reagent: firstly, citric acid buffer solution: 2.1g of citric acid was added to 100mL of distilled water to prepare solution A, and 2.94g of sodium citrate was added to 100mL of distilled water to prepare solution B. Mixing solution A and solution B at a ratio of 1:1.32 or 1:1, adjusting pH to 4.2-4.5, and filtering the mixed solution with a 2.22 μm filter.

The sulfathiazole solution (1 mg/kg): after DMSO dissolution, the mixture was diluted with physiological saline to the desired concentration

A metformin hydrochloride solution (200 mg/kg): is prepared by normal saline

Establishment of C57BL/6N mouse type 2 diabetes model

80 mice were randomly divided into three groups:

general feed group (SD): and 12 animals are fed with common feed for three weeks, and then are intraperitoneally injected with citric acid buffer solution (the injection dosage is the same as that of the STZ solution described below), fasted for 12 hours before the citric acid buffer solution is injected, injected according to the same time period of 9:00am-10:00am for five days once a day, and fed after the citric acid buffer solution is injected for 2 hours. Then feeding with common feed for three weeks, and detecting fasting blood glucose of mice between 7:30pm and 9:00pm without water supply for 10 h.

High fat diet group (HFD): after feeding 12 animals with high-fat feed for three weeks, injecting citric acid buffer solution (the injection dosage is same as the STZ solution described below) into abdominal cavity, fasting before injecting citric acid buffer solution and not forbidding water for 12h, injecting according to the same time period of 9:00am-10:00am, injecting once a day and continuously for five days, and feeding after injecting citric acid buffer solution for 2 h. Then feeding with high fat feed for three weeks, and detecting fasting blood glucose of mice between 7:30pm and 9:00pm without water supply for 10 h. The purpose of this group was to demonstrate that the type 2 diabetes model was induced by high fat diet in combination with STZ, but not by high fat diet alone.

Type 2 diabetes Model group (Model): 56 animals were fed with high-fat diet for three weeks, and then were administered with 50mg/kg of STZ solution (STZ dissolved in citric acid buffer solution and prepared into 5% strength solution) intraperitoneally. And (3) fasting is carried out for 12 hours before the injection of the STZ solution, injection is carried out according to the same time period of 9:00am-10:00am, once every day and five days of continuous injection, and the newly prepared STZ solution is fed after being injected for 2 hours, so that the injection is finished within 30 min. And then feeding the mice with high-fat feed for three weeks, detecting fasting plasma glucose of the mice between 7:30pm and 9:00pm, selecting mice with Model group fasting plasma glucose of more than 15.0mmol/L, feeding the mice with common feed for one week, detecting the fasting plasma glucose again, considering that the mice with fasting plasma glucose still of more than 15.0mmol/L are successful type 2 diabetes models, selecting 18 mice as type 2 diabetes models, and feeding all the mice with the common feed after formal experiments begin.

18 mice successfully modeled were randomly divided into 3 groups, 12 common-diet mice were randomly divided into 2 groups, 12 high-fat-diet mice were divided into 2 groups, 7 groups were used in total, and 6 mice were fed with common diet after the full-scale start of the experiment. The specific grouping and administration conditions were as follows:

model group: injecting equal volume of normal saline into the abdominal cavity;

model + Metformin set: intragastric administration of 200mg/kg metformin hydrochloride;

model + sulfothiazole (Sulfathiazole) group: 1mg/kg of sulfathiazole compound is injected into the abdominal cavity;

SD (normal feed) group: injecting equal volume of normal saline into the abdominal cavity;

SD + Sulfathiazole group: 1mg/kg of sulfathiazole compound is injected into the abdominal cavity;

HFD (high fat diet) group: injecting equal volume of normal saline into the abdominal cavity;

HFD + sulfothiazole (Sulfathiazole) group: 1mg/kg of sulfathiazole compound is injected into the abdominal cavity;

all mice were dosed once daily for 13 weeks, during which time the growth and life of the mice were observed and weekly changes in body weight and blood glucose were measured.

4. Detecting the index

4.1 weight and fasting glucose assay

Monitoring body weight and fasting blood glucose in the same time period (7:30pm-9:00pm) before and after administration, after fasting for 10h every week, weighing with balance, and recording body weight value; cutting the tail, properly taking blood, measuring the blood sugar content by a glucometer (glucose oxidase method), and reading and recording the blood sugar value.

4.2 detection of diabetic nephropathy-associated indicators

At week 11 of administration, mice were assayed for 24h urine volume, urine protein content, urine creatinine, and urea nitrogen content, as determined according to kit instructions.

4.3 detection of sugar tolerance and insulin tolerance by intraperitoneal injection

At the 12 th week of administration, insulin tolerance test was performed, after fasting for 6 hours without water deprivation, the blood glucose content was measured with a glucometer as the blood glucose level at 0min (BG0), then 0.3U/kg insulin was intraperitoneally injected, the blood glucose levels at 15, 30, 60, and 120min (BG15, BG30, BG60, and BG120) after insulin injection were measured and recorded, respectively, the IPITT curve was plotted with Graphpad prism software, and the total area under the curve AUC of 0-120min blood sampling period was calculated.

At the 12 th week of administration, glucose tolerance test was performed, and mice of each group were fasted for 12 hours without water deprivation, and the next day blood glucose content was measured with a glucometer as the blood glucose level at 0min (BG0), followed by intraperitoneal injection of 1g/kg glucose, blood glucose levels at 15, 30, 60, and 120min (BG15, BG30, BG60, and BG120) after the recording of the injected glucose were measured, respectively, an IPGTT curve was plotted, and the total area under the curve AUC of the blood sampling period of 0-120min was calculated.

5. Dissected mice and preparation of specimens

At the end of 13 weeks after administration, after mice are fasted for 12 hours, the eyeballs are picked up to take blood, the whole blood added with the anticoagulant is kept still at 4 ℃ for 2 days, supernatant plasma is sucked, then the supernatant is centrifugally sucked at 4 ℃ for 1000g for 10min, and split charging (avoiding repeated freeze thawing) is carried out, and the split charging is carried out and the split charging is stored in a refrigerator at-80 ℃ for freezing and storage for standby. Pancreatic tissue, liver tissue, kidney tissue, heart tissue and spleen tissue are taken, placed on ice, and fixed in 4% paraformaldehyde solution (more than 24h) after being washed clean by physiological saline, and stored in a refrigerator at 4 ℃ for later use. Dehydrating with gradient alcohol, and making into conventional paraffin embedding and section; after hematoxylin-eosin staining and mounting, the images were observed and photographed under a microscope (magnification of 200 times).

5.1 detection of diabetic cardiovascular disease-related indices

And (3) measuring the contents of total cholesterol, triglyceride and low-density lipoprotein cholesterol in the plasma according to the kit instruction.

5.2 glycated hemoglobin assay

The content of glycated hemoglobin in mice was measured according to the kit instructions.

5.3 detection of Fasting Serum Insulin (FINS) content

And (3) measuring the fasting serum insulin content of the mouse, and measuring the FINS content in the serum according to the INSELISA kit instruction of the mouse.

6. Data processing

The experimental data were statistically analyzed using SPSS23.0 software, the data are expressed as (x. + -.s), and the comparisons between groups were pairwise compared using One-way ANOVA and LSD-t. p <0.05 was considered statistically significant.

7. Results of the experiment

7.1 Effect of Sulfathiazole on weight and blood glucose in type 2 diabetic mice

As shown in fig. 1, the body weights of the mice in the SD, SD + sulfothiazole, HFD and HFD + sulfothiazole groups continued to be higher than the Model group, but there was no significant difference between the groups; there was no significant difference in body weight between the Model + sulfothiazole group, the Model + Metformin group and the Model group.

As shown in fig. 2, blood glucose of the SD, SD + sulfothiazole, HFD and HFD + sulfothiazole mice was always maintained within the normal range (<7 mmol/L); the blood glucose persistence of Model group mice was significantly higher than that of SD group (### P < 0.001); from week 1 treatment initiation to week 13, blood glucose in mice in the Model + Sulfathiazole group was significantly lower than in the Model group at weeks 3, 4, 8, 9, 11, 12, and 13 (P < 0.05;. P < 0.01;. P < 0.001); mice in the Model + Metformin group were significantly lower in blood glucose than the Model group except for weeks 5 and 10 (P < 0.05;. P < 0.01;. P < 0.001); from the experimental results of blood sugar change, the Sulfathiazole can obviously reduce the blood sugar of mice with type 2 diabetes, has the blood sugar reducing effect 200 times higher than that of metformin, and can be used for preparing the medicament for treating diabetes.

7.2 Effect of Sulfathiazole on the Kidney of type 2 diabetic mice

Urine was measured in the mice of each group at week 11 of treatment, and as shown in FIG. 3, urine protein of the SD group mice was substantially identical to that of the SD + Sulfathiazole group, and was lower than that of the Model group (Model group vs. SD group # # # P < 0.001); the contents of the HFD group and the HFD + Sulfathiazole group urine protein are basically consistent and are lower than those of the Model group; the urinary protein of mice in the Model + Sulfahiazole group is lower than that of mice in the Model group, which indicates that the Sulfahiazole can reduce the urinary protein content of mice with type 2 diabetes; the Model + Metformin group mice had slightly higher urine protein than the Model group. As shown in fig. 4, there was no difference in urine protein content between the SD group, SD + sulfothiazole group, HFD group and HFD + sulfothiazole group; the content of the urinary creatinine in the mice of the Model + Sulfathiazole group is lower than that in the Model group, which shows that the Sulfathiazole can reduce the content of the creatinine in the urine of the mice with type 2 diabetes; the mice in the Model + Metformin group had lower urinary creatinine content than the Model group. As shown in fig. 5, there was no difference in urea nitrogen content between the SD group, SD + sulfozole group and HFD + sulfozole group, but the urea nitrogen content of the HFD group was substantially the same as that of the Model group, indicating that high fat diet resulted in increased urea nitrogen in mice; the urea nitrogen content of the Model + Sulfathiazole group mice is lower than that of the Model group, which shows that the Sulfathiazole can reduce the urea nitrogen content of the type 2 diabetes mellitus mice; from the experimental results of fig. 3-5, we can see that the sulfathiazole can reduce the contents of urine protein, urine creatinine and urea nitrogen in the mice with type 2 diabetes, and further can figure out that the sulfathiazole can protect the renal function of the mice with diabetes and treat the renal disease caused by diabetes.

7.3 Effect of Sulfathiazole on insulin tolerance in type 2 diabetic mice

The mice in the above groups were subjected to insulin tolerance test at week 11 of treatment, and as shown in FIG. 6, within 0-120min, the blood glucose variation tendency of the mice in the SD group, SD + Sulfathiazole group, HFD group and HFD + Sulfathiazole group was substantially the same; at 0min, 15min, 30min, the blood glucose of the Model + Sulfathiazole group mice was significantly lower than the Model group (. P < 0.05;. P <0.01), indicating that the Model + Sulfathiazole group mice were more insulin sensitive than the Model group; the blood sugar of mice in the Model + Metformin group is lower than that of mice in the Model group at 0min, 15min and 30 min; within 60-120min, there was no substantial change in blood glucose in the Model + Sulfathiazole group, Model + Metformin group and Model group mice. As shown in fig. 7, the area under the 0-120min blood glucose change curve shows no significant difference between the SD group, SD + Sulfathiazole group, HFD group and HFD + Sulfathiazole group; the area under the blood sugar change curve of the Model group at 0-120min is larger than that of the SD group (## P < 0.001); the area under the 0-120min blood glucose change curve of the Model + Sulfathiazole group is smaller than that of the Model group (P < 0.05); the area under the 0-120min blood sugar change curve of the Model + Metformin group is smaller than that of the Model group, which shows that the sulfathiazole can improve the sugar metabolism of the type 2 diabetic mice, and also shows that the sulfathiazole can improve the sensitivity of insulin and improve the insulin resistance.

7.4 Effect of Sulfathiazole on glucose tolerance in type 2 diabetic mice

Glucose tolerance test is carried out on the mice in each group at the 11 th week of treatment, as shown in figure 8, within 0-120min, the blood sugar change trends of the SD group, the SD + Sulfathiazole group and the HFD group are basically consistent, and the blood sugar of the HFD + Sulfathiazole group is slightly higher than that of the HFD group at 30min and 60 min; within 0-30min, the blood sugar of the Model + Sulfathiazole group rises more slowly than that of the Model group, and at 0min, 15min, 30min and 60min, the blood sugar is obviously lower than that of the Model group (P < 0.05; P <0.01), which indicates that the Model + Sulfathiazole group has stronger regulating capability on the glucose in vivo than that of the Model group; the Model + Metformin group was lower in blood glucose at 0min than the Model group (. P < 0.05). As shown in FIG. 9, the area under the 0-120min blood glucose change curve is substantially the same for the SD group, SD + Sulfathiazole group and HFD + Sulfathiazole group mice, and the HFD + Sulfathiazole group is higher than the HFD group; the area under the blood sugar change curve of the Model group at 0-120min is larger than that of the SD group (## P < 0.001); the area under the 0-120min blood sugar change curve of the Model + Sulfathiazole group is smaller than that of the Model group (P <0.01), and the area under the 0-120min blood sugar change curve of the Model + Metformin group is smaller than that of the Model group, which shows that the Sulfathiazole can improve the glucose metabolism and regulate the glucose balance in vivo.

7.5 Effect of Sulfathiazole on the cardiovascular System of type 2 diabetes

As shown in fig. 10, there was no difference in triglyceride content among the mice of SD group, SD + Sulfathiazole group, HFD group, and HFD + Sulfathiazole group; the triglyceride content of mice in the Model + Sulfathiamazole group is lower than that of mice in the Model group, which indicates that the Sulfathiamazole can reduce the triglyceride content of mice with type 2 diabetes; the triglyceride content of Model + Metformin group mice was slightly higher than that of Model group. As shown in fig. 11, there was no difference in the low density lipoprotein cholesterol levels among the SD, SD + sulfothiazole, HFD and HFD + sulfothiazole mice; the low-density lipoprotein cholesterol content of the Model + Sulfahiazole group mice is slightly lower than that of the Model group, which shows that the Sulfahiazole can reduce the low-density lipoprotein cholesterol content of the type 2 diabetes mellitus mice; the Model + Metformin group mice had lower LDL cholesterol levels than the Model group. As shown in fig. 12, the total cholesterol levels of the SD, SD + Sulfathiazole, HFD and HFD + Sulfathiazole mice were substantially the same; the content ratio of total cholesterol of mice in the Model + sulfothiazole group is more than that in the Model group; the total cholesterol level in Model + Metformin mice was slightly lower than in the Model group. The sulfonamide compound sulfathiazole can reduce the content of triglyceride and total cholesterol of a type 2 diabetic mouse, regulate the blood fat level, and improve the lipid metabolism of the diabetic mouse, so that the sulfonamide compound sulfathiazole can treat diabetic cardiovascular complications.

EXAMPLE II Effect of sulfadiazine treatment of diabetes mellitus and complications thereof in mice

1. The animal feeding mode is as shown in the first example:

2. drugs and reagents:

the medicines used in the invention are as follows: sulfadiazine (Sulfadiazine); metformin hydrochloride (Metformin hydrochloride, Metformin); streptozotocin (streptazocin, STZ).

Preparation of a reagent: firstly, citric acid buffer solution: 2.1g of citric acid was added to 100mL of distilled water to prepare solution A, and 2.94g of sodium citrate was added to 100mL of distilled water to prepare solution B. Mixing solution A and solution B at a ratio of 1:1.32 or 1:1, adjusting pH to 4.2-4.5, and filtering the mixed solution with a 2.22 μm filter.

Preparation of sulfadiazine solution (1 mg/kg): dissolving DMSO, and diluting with normal saline to desired concentration

The preparation method comprises the following steps of (1) preparing a metformin hydrochloride solution (200 mg/kg): is prepared by normal saline

Establishment of C57BL/6N mouse type 2 diabetes model

Type 2 diabetes Model group (Model): 56 animals were fed with high-fat diet for three weeks, and then were administered with 50mg/kg of STZ solution (STZ dissolved in citric acid buffer solution and prepared into 5% strength solution) intraperitoneally. And (3) fasting is carried out for 12 hours before the injection of the STZ solution, injection is carried out according to the same time period of 9:00am-10:00am, once every day and five days of continuous injection, and the newly prepared STZ solution is fed after being injected for 2 hours, so that the injection is finished within 30 min. And then feeding the mice with high-fat feed for three weeks, detecting fasting plasma glucose of the mice between 7:30pm and 9:00pm, selecting mice with Model group fasting plasma glucose of more than 15.0mmol/L, feeding the mice with common feed for one week, detecting the fasting plasma glucose again, considering that the mice with fasting plasma glucose still of more than 15.0mmol/L are successful type 2 diabetes models, selecting 18 mice as type 2 diabetes models, and feeding all the mice with the common feed after formal experiments begin.

The 18 successfully modeled type 2 diabetes mice are randomly grouped into 3 groups, and the normal mice are 1 group and are totally divided into 4 groups, each group comprises 6 mice, and the specific grouping and administration conditions are as follows:

model group: perfusing normal saline with the same volume as the stomach;

model + metformin set: intragastric administration of 200mg/kg metformin hydrochloride;

model + Sulfadiazine (Sulfadiazine) group: gavage 1mg/kg sulfadiazine compound;

SD (normal feed) group: perfusing normal saline with the same volume as the stomach;

all mice were dosed once daily for 3 weeks, during which time the growth and life of the mice were observed and weekly changes in body weight and blood glucose were measured.

4. Detecting the index

4.1 weight and fasting glucose assay

Monitoring body weight and fasting blood glucose in the same time period (7:30pm-9:00pm) before and after administration, after fasting for 10h every week, weighing with balance, and recording body weight value; cutting the tail, properly taking blood, measuring the blood sugar content by a glucometer (glucose oxidase method), and reading and recording the blood sugar value.

5. The experimental results are as follows:

5.1 Effect of sulfadiazine on weight and blood glucose in type 2 diabetic mice

As shown in FIG. 13, there was no significant difference in body weight among the mice of the SD group, Model + metformin group, and Model + Sulfadiazine group. As shown in fig. 14, blood glucose in SD group mice was always maintained within the normal range (<7 mmol/L); the blood glucose persistency of Model group mice was significantly higher than that of SD group (# # P < 0.01; # # P < 0.001); at both week 1 and week 3, mice in the Model + Sulfadiazine group had significantly lower blood glucose than the Model group (. P < 0.05); at weeks 1, 2, and 3, blood glucose was significantly lower in the Model + metformin group mice than in the Model group (. P < 0.01;. P <0.001), indicating that sulfadiazine had therapeutic effects in type 2 diabetic mice.

EXAMPLE III Effect of sulfamethazine on the treatment of diabetes mellitus and its complications in mice

1. The animal feeding mode is as shown in the first example:

2. drugs and reagents:

the medicines used in the invention are as follows: sulfamethazine (Sulfamerazine); metformin hydrochloride (Metformin hydrochloride, Metformin); streptozotocin (streptazocin, STZ).

Preparation of a reagent: firstly, citric acid buffer solution: 2.1g of citric acid was added to 100mL of distilled water to prepare solution A, and 2.94g of sodium citrate was added to 100mL of distilled water to prepare solution B. Mixing solution A and solution B at a ratio of 1:1.32 or 1:1, adjusting pH to 4.2-4.5, and filtering the mixed solution with a 2.22 μm filter.

Mazeflo (1 mg/kg): dissolving DMSO, and diluting with normal saline to desired concentration

The preparation method comprises the following steps of (1) preparing a metformin hydrochloride solution (200 mg/kg): is prepared by normal saline

Establishment of C57BL/6N mouse type 2 diabetes model

Type 2 diabetes Model group (Model): 56 animals were fed with high-fat diet for three weeks, and then were administered with 50mg/kg of STZ solution (STZ dissolved in citric acid buffer solution and prepared into 5% strength solution) intraperitoneally. And (3) fasting is carried out for 12 hours before the injection of the STZ solution, injection is carried out according to the same time period of 9:00am-10:00am, once every day and five days of continuous injection, and the newly prepared STZ solution is fed after being injected for 2 hours, so that the injection is finished within 30 min. And then feeding the mice with high-fat feed for three weeks, detecting fasting plasma glucose of the mice between 7:30pm and 9:00pm, selecting mice with Model group fasting plasma glucose of more than 15.0mmol/L, feeding the mice with common feed for one week, detecting the fasting plasma glucose again, considering that the mice with fasting plasma glucose still of more than 15.0mmol/L are successful type 2 diabetes models, selecting 18 mice as type 2 diabetes models, and feeding all the mice with the common feed after formal experiments begin.

The 18 successfully modeled type 2 diabetes mice are randomly grouped into 3 groups, and the normal mice are 1 group and are totally divided into 4 groups, each group comprises 6 mice, and the specific grouping and administration conditions are as follows:

model group: perfusing normal saline with the same volume as the stomach;

model + metformin set: intragastric administration of 200mg/kg metformin hydrochloride;

model + Sulfamerazine (sulfamethazine) group: gavage 1mg/kg of a sulfamethazine compound;

SD (normal feed) group: perfusing normal saline with the same volume as the stomach;

all mice were dosed once daily for 3 weeks, during which time the growth and life of the mice were observed and weekly changes in body weight and blood glucose were measured.

4. Detecting the index

4.1 weight and fasting glucose assay

Monitoring body weight and fasting blood glucose in the same time period (7:30pm-9:00pm) before and after administration, after fasting for 10h every week, weighing with balance, and recording body weight value; cutting the tail, properly taking blood, measuring the blood sugar content by a glucometer (glucose oxidase method), and reading and recording the blood sugar value.

5. The experimental results are as follows:

5.1 Effect of sulfamethazine on weight and blood glucose in type 2 diabetic mice

As shown in FIG. 15, there was no significant difference in body weight among the mice of the SD group, Model + metformin group, and Model + Sulfamerazine group. As shown in fig. 16, blood glucose in SD group mice was always maintained within the normal range (<7 mmol/L); the blood glucose persistency of Model group mice was significantly higher than that of SD group (# # P < 0.01; # # P < 0.001); at weeks 1 and 2, the blood glucose of the Model + Sulfamerazine group mice was lower than the Model group, and at week 3, the blood glucose of the Model + Sulfamerazine group mice was significantly lower than the Model group (. P < 0.05); at weeks 1, 2, and 3, blood glucose was significantly lower in the Model + metformin group mice than in the Model group (. P < 0.05;. P < 0.01;. P <0.001), indicating that sulfamethazine had a therapeutic effect in type 2 diabetic mice.

Conclusion

In conclusion, the sulfonamide compound sulfathiazole can improve the glucose metabolism of type 2 diabetes mice, improve the sensitivity of insulin, improve insulin resistance, and obviously reduce the blood sugar of the type 2 diabetes mice, and the blood sugar reducing effect of the sulfonamide compound sulfathiazole is 200 times or more higher than that of metformin; the blood sugar reducing effect of the sulfonamide compound is also remarkable in sulfadiazine and sulfamethazine; meanwhile, the sulfanilamide compound sulfathiazole can reduce the contents of urine protein, urine creatinine and urea nitrogen of a type 2 diabetes mouse, which indicates that the sulfanilamide compound sulfathiazole can protect the renal function of the diabetes mouse; the sulfanilamide compound sulfathiazole can reduce the content of triglyceride and total cholesterol of a type 2 diabetes mouse, regulate the blood fat level and improve the lipid metabolism of the diabetes mouse. . The sulfa compound sulfadiazine can obviously reduce the blood sugar of a type 2 diabetic mouse and has a therapeutic effect on the type 2 diabetic mouse; the sulfanilamide compound sulfamethazine can obviously reduce the blood sugar of a type 2 diabetes mouse, and has a treatment effect on the type 2 diabetes mouse. The sulfonamide compound can be used for treating cardiovascular complications of diabetes and can be clinically popularized and applied.