CN107320471B

CN107320471B - Cycloserine pharmaceutical composition for treating tuberculosis and application thereof

Info

Publication number: CN107320471B
Application number: CN201710668428.7A
Authority: CN
Inventors: 牛文一; 李明瑛; 万勇敢; 黄健; 张力公
Original assignee: Hunan Huize Bio Pharmaceutical Co ltd
Current assignee: HUNAN HUIZE BIO-PHARMACEUTICAL Co.,Ltd.
Priority date: 2017-08-08
Filing date: 2017-08-08
Publication date: 2020-10-09
Anticipated expiration: 2037-08-08
Also published as: CN107320471A

Abstract

The invention discloses a cycloserine medicinal composition for treating tuberculosis and application thereof, belonging to the technical field of medicines. The pharmaceutical composition takes cycloserine and tranexamic acid as active ingredients, the dosage ratio of the tranexamic acid to the cycloserine is (2.5-10.0):1, the two components can not only quickly kill mycobacterium tuberculosis by exerting synergistic action, but also recover the activity of tissues eroded by the mycobacterium tuberculosis to a certain extent, and is suitable for treating various tuberculosis.

Description

Cycloserine pharmaceutical composition for treating tuberculosis and application thereof

Technical Field

The invention belongs to the technical field of western medicine active compounds, and particularly relates to a cycloserine medicinal composition for treating tuberculosis and application thereof.

Background

Tuberculosis, one of the major public health problems facing the world today, is the causative agent Mycobacterium tuberculosis (Mtb), a genus of Mycobacterium. In recent years, the tuberculosis epidemic has been increasing due to the emergence of Human Immunodeficiency Virus (HIV) in combination with mycobacterium tuberculosis infection, large-scale population mobility and other problems. At present, about 900 million people develop into active tuberculosis every year around the world, and 200-300 million people die from tuberculosis. It is estimated that about 1/3 of the population is infected with Mtb worldwide, most of which are latent tuberculosis infections (LTBI), i.e. a state in which the body's immune system is able to control the replication of tubercle bacillus without developing into tuberculosis, but without completely eliminating it. At this time, the tubercle bacillus is in a latent state, and in the case of low immunity, the tubercle bacillus can be replicated again and develop into active tuberculosis to become a new infection source. The infected people have a risk of developing active tuberculosis of about 5-10% throughout life, and if the infected people are simultaneously infected by HIV, the probability is up to 10% per year, which is far higher than that of HIV-negative people. Epidemiological investigations have shown that about 85% to 90% of the newly diagnosed active tubercle bacilli test positive LTBI evolves.

Currently, the treatment of tuberculosis employs a formulation of a combination of agents recommended by the U.S. public health agency, including first two months with isoniazid, rifampin, pyrazinamide, and ethambutol, followed by four months with isoniazid and rifampin alone. For patients infected with AIDS, the use of the pharmaceutical composition needs to be delayed to seven months. For patients infected with multi-drug resistant tuberculosis, the pharmaceutical composition is added with other second-line agents, such as streptomycin, kanamycin, amikacin, capreomycin, ethionamide, cycloserine, ciprofloxacin and ofloxacin. Such combination therapies for patients resistant to multi-drug tuberculosis (typically over a 2 year course of treatment) generally have lower activity and higher side effects than the first-line drugs currently on the market. In addition, the drugs have strong toxic and side effects, such as numbness of limbs caused by isoniazid, streptomycin has toxicity to auditory nerve and vestibular nerve, cycloserine has large toxic reaction to nervous system, gastrointestinal tract fever resistance and other toxic and side effects. Moreover, most anti-tuberculosis drugs have toxic and side effects on the liver in different degrees, and are difficult to accept by patients with high price.

Therefore, there is an urgent need for new antitubercular drugs with high activity to achieve the purpose of shortening the treatment time and reducing the toxic and side effects. At present, there is no report on tranexamic acid for inhibiting and killing mycobacterium tuberculosis, and there is no report on tranexamic acid combined with cycloserine for treating tuberculosis.

Disclosure of Invention

In view of the problems of poor curative effect and large toxic and side effect of cycloserine on tuberculosis, the invention aims to find a treatment method which has good curative effect and small adverse reaction and can be maintained for long-term use, thereby providing the pharmaceutical composition for treating tuberculosis by taking cycloserine and tranexamic acid as active ingredients.

The inventor has unexpectedly found in clinical application that when a patient with gastrorrhagia simultaneously suffers from tuberculosis and stops bleeding through tranexamic acid injection after hospitalization, the biological activity of cycloserine in killing mycobacterium tuberculosis can be remarkably enhanced. Aiming at the discovery, the inventor further designs in vitro and in vivo tests, and finally discovers that the combined medicine of tranexamic acid and cycloserine in a certain proportion range has the synergistic effect of quickly inhibiting and killing mycobacterium tuberculosis.

Therefore, the inventor provides a pharmaceutical composition for treating tuberculosis on the basis of taking cycloserine and tranexamic acid as active ingredients, thereby achieving the aim of the invention. The specific technical scheme is as follows:

a pharmaceutical composition for treating tuberculosis is prepared from active ingredients and pharmaceutically acceptable adjuvants, wherein the active ingredients comprise tranexamic acid and cycloserine. It should be noted that the compound tranexamic acid used in the present invention is trans-4-aminomethyl cyclohexanecarboxylic acid, and cycloserine is dextro-4-amino-3-tetrahydroisooxazolone.

Further preferably, the pharmaceutical composition for the treatment of tuberculosis as described above, wherein the active ingredients consist of tranexamic acid and cycloserine.

Still further preferably, the pharmaceutical composition for treating tuberculosis as described above, wherein the ratio of the amount of tranexamic acid to the amount of cycloserine in the active ingredients is (2.5-10.0): 1.

Still further preferably, the pharmaceutical composition for treating tuberculosis as described above, wherein the ratio of the amount of tranexamic acid to the amount of cycloserine in the active ingredients is (2.5-5.0): 1.

Still further preferably, the pharmaceutical composition for treating tuberculosis as described above is a solid oral preparation, and each unit preparation contains 500mg of tranexamic acid and 50-200mg of cycloserine. Still further preferably, the unit preparation contains 500mg of tranexamic acid and 100 mg of cycloserine and 200mg of cycloserine.

Still further preferably, the pharmaceutical composition for treating tuberculosis as described above, wherein the solid oral preparation comprises tablet, capsule, granule, and dripping pill.

Based on the research results of the inventor and the common technical knowledge of the technicians in the field, the invention also provides a pharmaceutical application, namely, the application of tranexamic acid in preparing the drugs for treating tuberculosis; or the application of the active ingredient composition consisting of tranexamic acid and cycloserine in the preparation of the drugs for treating tuberculosis; further preferably, the invention also provides application of an active ingredient composition consisting of tranexamic acid and cycloserine in preparing a medicament for treating tuberculosis.

Compared with the prior art, the pharmaceutical composition provided by the invention takes cycloserine and tranexamic acid as active ingredients, and the cycloserine and tranexamic acid can rapidly kill mycobacterium tuberculosis by exerting a synergistic effect, and can recover the activity of tissues eroded by the mycobacterium tuberculosis to a certain extent, similar to severe tuberculosis patients with hemoptysis and bedridden degree. In addition, the invention greatly shortens the course of treatment of tuberculosis patients and enhances the curative effect. Most importantly, when the medicine is used for tuberculosis patients, only one fourth to one third of the original clinical dosage needs to be taken, so that the medicine has little damage to the liver and the kidney.

Detailed Description

The invention will be further described with reference to specific embodiments, and the advantages and features of the invention will become apparent as the description proceeds. It should be understood that the illustrated embodiments are exemplary only, and are not intended to limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit of the invention, and it is intended to cover all such changes and modifications as fall within the scope of the invention.

Example 1: experimental study on effect of tranexamic acid combined with cycloserine on mycobacterium tuberculosis

1. Preparing an improved Luo culture medium inclined plane: 2.4g/L potassium dihydrogen phosphate, 0.24g/L magnesium sulfate, 0.6g/L magnesium citrate, 7.2g/L sodium L-glutamate, 0.4g/L malachite green and 39.0g/L potato starch. The preparation method comprises the following steps: 49.84g of the components are weighed according to the formula, 12ml of glycerol is absorbed, the components are heated, stirred and dissolved in 600ml of distilled water, boiled for 5-10 minutes, autoclaved at 121 ℃ for 15 minutes and taken out. Cooling to about 55 deg.C, adding sterile and stirred whole egg solution 1000ml under aseptic condition, mixing (to avoid generation of bubbles), subpackaging with 20mm × 150mm test tubes with 8ml each, placing on long inclined plane, and steaming at 85 deg.C for 50 min.

2. The preparation of the mycobacterium tuberculosis suspension comprises weighing 0.1mg of mycobacterium tuberculosis H37RV standard bacteria (ATCC: 27294) colony, adding 0.5mL of mixed diluent of physiological saline and Sutong culture solution (volume ratio is 1: 3) to inoculate on the slope of the improved Roche medium, culturing at 37 deg.C for 18d, collecting the colony growing well on the improved Roche medium, adding a small amount of physiological saline containing 0.05% Tween 80, grinding uniformly, diluting to obtain white and uniformly turbid bacteria solution, counting under microscope, adjusting bacteria concentration to 3 × 10⁷one/mL (for testing).

3. Test grouping and index data acquisition: preparing the improved Roche culture medium into a drug-containing plate and a drug-free plate respectively according to the method, namely: a placebo containing no drug; cycloserine group (cycloserine 5, 10 mg/mL); contains tranexamic acid groups with different concentrations (40, 20, 10, 5 mg/mL); a combination drug group containing tranexamic acid (40, 20, 10, 5mg/mL) at different concentrations based on cycloserine (5 mg/mL). Each group of media contained 15mL per plate. Inoculating 0.1mL of the bacterial suspension prepared by the method on the surface of each medicine-containing plate and each non-medicine-containing plate, setting 6 plates for each concentration, culturing at 37 ℃, observing for 1 time every 5 days, and counting the number of bacterial colonies of each plate by taking the surface of a blank control group mycobacterium tuberculosis overgrow culture medium as a termination culture standard after 40 days.

4. And (3) analyzing test results: the test statistics results in table 1 show that tranexamic acid shows a certain bacteriostatic action with the increase of the dosage, the inhibition and killing action on H37RV standard bacteria is incomplete when the cyclic serine is 5mg/mL, but the combined use of the tranexamic acid with the concentration of 20 mg/mL and 40mg/mL shows a very significant bactericidal action.

TABLE 1 comparison of the number of colonies of Mycobacterium tuberculosis H37RV each containing a drug group

Example 2 Tranexamic acid in combination with cycloserine Effect on tuberculosis mice

1. Replication of mouse tuberculosis model: 32 clean KM mice, 8 weeks old, 18-22g in body mass, male and femaleThe specific method for infecting mouse with human Mycobacterium tuberculosis H37RV standard strain comprises collecting 0.4mL of a strain containing 6.6 × 10⁵Injecting the mycobacterium tuberculosis H37RV bacterial suspension into tail veins of mice so as to establish a tuberculosis model of the mice.

2. Grouping and administration: tuberculosis model mice are randomly divided into four groups according to a male-female half mode: CK, CS, TXA, CS + TXA groups, 8 mice per group. Beginning on day 2 when the mice are infected with the mycobacterium tuberculosis H37RV, the mice of each administration group are perfused with the corresponding drug for 2 times/d for 6 weeks on the basis of the body mass of the mice; the CK group was gavaged with an equal volume of saline 2 times/day for 6 weeks.

3. Acquiring test index data: after completion of the administration, animals were sacrificed, thymuses of mice were harvested, and after residual blood was blotted with filter paper, the thymus indices were obtained by weighing (mg), dividing by the body weight (g) of the mice, and multiplying by 10. Lung and spleen colony counts: the lung and spleen of the mouse are ground, diluted and inoculated on an improved Roche plate culture medium, and cultured for 4 weeks at 37 ℃ for colony counting.

4. And (3) analyzing test results: as can be seen from the test results in Table 2, the thymus weight index of mice in each administration group is improved to a certain extent after 6 weeks of treatment compared with the CK group, and particularly, the weight index of mice in the CS + TXA group is remarkably different from that in the CK group or 2 single drug groups (P is less than 0.05). In addition, the lung and spleen colony numbers of the CS + TXA group mice are significantly lower than those of the CK group, the CS group and the TXA group, and all have significant statistical difference (P < 0.05). The results of observing the animal organs show that the liver and the kidney of the CS + TXA group mice have no obvious change.

Table 2 comparison of the test indices of the groups of mice (n ═ 8)

Compared with the CK group, the data of the data are compared,^*p is less than 0.05; in comparison with the CS group,^#p is less than 0.05; in comparison with the TXA group,^￥P＜0.05。

Claims

1. the application of tranexamic acid in preparing medicine for killing mycobacterium tuberculosis is disclosed.

2. The application of the active ingredient composition consisting of tranexamic acid and cycloserine according to the weight ratio of (2.5-10.0):1 in the preparation of the medicine for killing mycobacterium tuberculosis.