阅读说明：本技术 甲磺酸萘莫司他在治疗衣原体生殖道感染的应用方法 (Application method of nafamostat mesylate in treatment of chlamydia genital tract infection ) 是由 陈建林 彭亮 张宏波 沈宏伟 唐玲丽 胡紫昊 赵宇婕 刘珊珊 鲁琼 于 2020-06-15 设计创作，主要内容包括：本发明涉及生物医药技术领域,尤其为甲磺酸萘莫司他在治疗衣原体生殖道感染的应用方法,包括以下步骤：A、选定实验材料；B、制定实验方法；C、观察实验结果；D、得出实验结论。本发明提供甲磺酸萘莫司他在治疗衣原体生殖道感染的新用途,尤其涉及甲磺酸萘莫司他制备治疗衣原体生殖道感染的药物方面的应用,以及甲磺酸萘莫司他在制备预防由衣原体生殖道感染引起的输卵管梗阻、输卵管积水、输卵管性不孕的药物方面的应用,而且通过体外细胞实验、动物实验等多个方面证明了甲磺酸萘莫司他可有效地治疗衣原体生殖道感染,进而可用于防治输卵管性不孕疾病。(The invention relates to the technical field of biological medicines, in particular to an application method of nafamostat mesilate in treating chlamydia genital tract infection, which comprises the following steps: A. selecting an experimental material; B. making an experimental method; C. observing an experimental result; D. and (5) obtaining an experimental conclusion. The invention provides a new application of nafamostat mesilate in treating chlamydia genital tract infection, in particular to an application of nafamostat mesilate in preparing a medicament for treating chlamydia genital tract infection, and an application of nafamostat mesilate in preparing a medicament for preventing fallopian tube obstruction, hydrosalpinx and tubal infertility caused by chlamydia genital tract infection.)

1. An application method of nafamostat mesilate in treating chlamydia genital tract infection is characterized in that: the method comprises the following steps:

A. selecting an experimental material;

B. making an experimental method;

C. observing an experimental result;

D. and (5) obtaining an experimental conclusion.

2. The method of claim 1 for the treatment of chlamydial genital tract infections with nafamostat mesylate, wherein the method comprises: the experimental materials in the step A comprise any several of strains, animals, cells and main reagents.

3. The method of claim 1 for the treatment of chlamydial genital tract infections with nafamostat mesylate, wherein the method comprises: the experimental method in the step B is specifically divided into three types,

the inhibition effect of nafamostat mesylate on chlamydia infection genital tract epithelial cells is achieved;

② the influence of nafamostat mesylate on the genital tract of mice infected with chlamydia;

③ the influence of the vaginal perfusion of nafamostat mesylate on the upper and lower genital tract cytokines of the mouse infected by the chlamydia.

Technical Field

The invention relates to the technical field of biomedicine, in particular to an application method of nafamostat mesilate in treating chlamydia genital tract infection.

Background

Chlamydia (Chlamydia) is a group of very small, non-motile, intracellularly growing microorganisms, and urogenital infections caused by them are the most common sexually transmitted diseases worldwide, with about 1.272 million new patients worldwide per year according to WHO statistics^[1]. Since the clinical manifestations of chlamydial infections are nonspecific, especially among women infected, asymptomatic individuals account for approximately 80% of the population, and are therefore often missed in clinical diagnosis. If the patient is not diagnosed and treated in time, the patient is easy to develop chronic and persistent infection, which causes the injury, adhesion and scar formation of the genital tract tissue, and then causes various complications such as tubal infertility, pelvic inflammation, ectopic pregnancy and the like^[2-4]And seriously affects the female reproductive health. In addition, genital tract chlamydia infection increases the risk of cervical cancer and AIDS^[5,6]It brings a heavy burden to the family and society.

Currently, according to the consensus of diagnosis and treatment of chlamydial genital tract infection recommended by WHO, azithromycin and doxycycline are first-line drugs for treating chlamydial genital tract infection, and although most chlamydial genital tract infections can be cured by antibiotic therapy, reports of antibiotic treatment failure have been increasing in recent years due to the emergence of multidrug-resistant chlamydia^[7,8]. Due to the broad spectrum of broad-spectrum antibioticsThe widespread use and the general use of empirical therapy during diagnosis and treatment may lead to the emergence of more multidrug-resistant chlamydiae in the future^[9]. Therefore, before the emergence of effective chlamydia vaccines, it is urgent to find other effective drugs capable of treating chlamydia genital tract infection and preventing the occurrence of pathological damage to the fallopian tube.

Nafamostat mesylate (CAS: 82956-11-4) is a synthetic serine protease inhibitor that has been approved by the FDA for: amelioration of acute symptoms of pancreatitis, treatment of patients with Disseminated Intravascular Coagulation (DIC), anticoagulation during extracorporeal circulation of blood in patients who are hemorrhagic or who have a tendency to bleed^[10,11]. Pharmacological studies show that nafamostat mesylate has strong selective inhibition effects on trypsin, thrombin, kallikrein, plasmin, fibrin protease, active blood coagulation factors (XIIa, Xa and VIIa) and trypsin-like serine protease of the classical pathway of the complement system. Recent studies have shown that the compounds are also useful in the treatment of diseases in other areas: the compound can relieve lung injury induced by inflammatory factor by its antiinflammatory effect^[12](ii) a Can be used for treating ischemia reperfusion injury of kidney and brain^[13](ii) a Pancreatic cancer inhibition by down-regulation of NF- κ B pathway^[14]Colorectal cancer^[15]Growing of (3); growth inhibition of triple negative breast cancer by downregulation of the Smad2-TGF β pathway^[16](ii) a Can be used for treating reactive arthritis induced by chlamydia^[17]。

At present, the application of nafamostat mesylate in treating chlamydia genital tract infection and preventing complications such as fallopian tube obstruction, hydrosalpinx, fallopian tube infertility and the like is not reported.

Disclosure of Invention

The invention aims to provide an application method of nafamostat mesilate in treating chlamydia genital tract infection, which has the advantage of providing a new application of nafamostat mesilate in treating chlamydia genital tract infection and solves the problems in the background technology.

In order to achieve the purpose, the invention provides the following technical scheme: the application method of nafamostat mesylate in treating chlamydia genital tract infection comprises the following steps:

A. selecting an experimental material;

B. making an experimental method;

C. observing an experimental result;

D. and (5) obtaining an experimental conclusion.

Preferably, the experimental materials in the step A comprise any several of strains, animals, cells and main reagents.

Preferably, the experimental method in the step B is specifically divided into three,

the inhibition effect of nafamostat mesylate on chlamydia infection genital tract epithelial cells is achieved;

② the influence of nafamostat mesylate on the genital tract of mice infected with chlamydia;

③ the influence of the vaginal perfusion of nafamostat mesylate on the upper and lower genital tract cytokines of the mouse infected by the chlamydia.

Compared with the prior art, the invention has the following beneficial effects:

1. nafamostat mesylate was used for the first time to treat chlamydial genital tract infection;

2. because of the wide use of the traditional antibiotics, part of the germ-line chlamydiae generate mutation drug resistance, the traditional antibiotic therapy has the risk of treatment failure, but the technical scheme is not used clinically as the antibiotic, has low drug resistance and can replace the traditional antibiotic scheme to treat the acute stage chlamydia infection;

3. provides a new administration route for treating acute chlamydia infection, namely vaginal local administration, and compared with the traditional systemic antibiotic treatment, the administration route has the following advantages: the vaginal local drug application mode improves the local drug concentration of the focus, and the drug can more directly act on chlamydia and target cells infected by the chlamydia; secondly, the influence of the medicine on other organs is reduced by a local medicine application mode of the vagina, and the metabolic burden of organs such as the liver, the kidney and the like is reduced; and the mode of local vaginal administration also reduces the occurrence of flora imbalance after systemic administration of antibiotics.

Drawings

FIG. 1 is a graph showing the effect of nafamostat mesylate on the growth of chlamydia inclusion bodies according to the present invention;

(Note: the upper image is a fusion image of a bright field in which HeLa229 cells are present and red fluorescence in which chlamydia inclusion bodies are present)

FIG. 2 is a graph showing the effect of nafamostat mesylate on the number of chlamydia inclusions in accordance with the present invention;

(Note: P < 0.01X P <0.001, total 3 independent experiments, each time in different concentrations of the multiple holes)

FIG. 3 is a graph showing the effect of nafamostat mesylate on the diameter of chlamydia inclusion bodies according to the invention;

(Note: P <0.001, total 3 independent experiments, each experiment in different concentrations of the multiple holes)

FIG. 4 is a graph showing the change in vaginal bacterial load with time after infection with Chlamydia in different groups of mice according to the invention;

(Note: P < 0.05X P < 0.01X P <0.001, n ═ 5-7)

FIG. 5 is a graph showing the effect of nafamostat mesylate of the invention on cytokines in the upper and lower reproductive tracts of mice after infection;

(Note: P <0.05n ═ 5)

FIG. 6 is the structural formula of nafamostat mesylate of the invention;

FIG. 7 is a schematic flow chart of the method of the present invention.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Referring to fig. 1-7, a method of using nafamostat mesylate in the treatment of chlamydial genital tract infection comprising the steps of:

A. selecting an experimental material;

B. making an experimental method;

C. observing an experimental result;

D. and (5) obtaining an experimental conclusion.

The experimental materials in the step A comprise any several of strains, animals, cells and main reagents.

The experimental method in the step B is specifically divided into three types,

the inhibition effect of nafamostat mesylate on chlamydia infection genital tract epithelial cells is achieved;

② the influence of nafamostat mesylate on the genital tract of mice infected with chlamydia;

③ the influence of the vaginal perfusion of nafamostat mesylate on the upper and lower genital tract cytokines of the mouse infected by the chlamydia.