CN112156088B

CN112156088B - Application of compound in preparation of mycobacterium tuberculosis inhibitor

Info

Publication number: CN112156088B
Application number: CN202011196370.9A
Authority: CN
Inventors: 赵刚; 吴雪英; 余丽花; 张梦婷; 黄佩闻; 周力华
Original assignee: Jianmin Pharmaceutical Groups Corp ltd
Current assignee: Jianmin Pharmaceutical Groups Corp ltd
Priority date: 2020-10-30
Filing date: 2020-10-30
Publication date: 2021-08-31
Anticipated expiration: 2040-10-30
Also published as: CN112156088A

Abstract

The invention discloses application of a compound in preparing a mycobacterium tuberculosis resistant medicament. The inhibition effect of the compound on mycobacterium tuberculosis is verified through an in vitro bacteriostatic test, a guinea pig lymphoid tuberculosis animal model is established by injecting mycobacterium tuberculosis liquid subcutaneously, the influence of the compound on the number and weight of swollen lymph nodes of the model animal is investigated, and the result shows that the compound can reduce the number and weight of the swollen lymph nodes and is dose-dependent, which indicates that the compound also has a better effect in vivo. The structural formula of the compound related by the invention is as follows.

Description

Application of compound in preparation of mycobacterium tuberculosis inhibitor

Technical Field

The invention belongs to the field of pharmacy, and relates to a new pharmaceutical application of a compound, in particular to a new application in preparation of a mycobacterium tuberculosis inhibitor.

Background

A traditional Chinese medicine with the effects of dissipating stagnation, reducing swelling, removing blood stasis and relieving pain is collected from the section of 2015 edition of Chinese pharmacopoeia, and the medicine is named as Xiaojin capsule and is obtained by modifying the dosage form based on Xiaojin pills. The traditional Chinese medicine is prepared from 10 traditional Chinese medicines such as artificial musk, semen momordicae, radix aconiti agrestis, resina liquidambaris, frankincense, myrrh, trogopterus dung, angelica sinensis, earthworm and Chinese ink, is a classic and famous traditional Chinese medicine prescription for treating cellulitis, and has the effects of eliminating stagnation, reducing swelling, removing blood stasis and relieving pain. It is mainly used for treating scrofula, goiter, mammary cancer and nodules of breast caused by phlegm-qi stagnation, with symptoms of skin or one or more parts of mass under skin, push movement, or bone and bone joint swelling, skin color unchanged, swelling, hardness and pain. Has good curative effect on thyroid enlargement, benign prostatic hyperplasia, cystic hyperplasia of mammary glands, chronic pelvic inflammatory disease mass, nodular fasciitis and the like which are the names of western medicine diseases.

Lymphoid tuberculosis, called scrofula in traditional Chinese medicine, is a toxic mass tissue which is formed by phlegm, toxic heat and toxic coagulation and is reflected on the muscle surface, and is considered to be infected on a primary focus by tubercle bacillus by western medicine. A large amount of clinical data prove that the small gold capsules have good curative effect on the scrofula.

In the research on effector substances and action mechanisms of the traditional Chinese medicine, the applicant carries out systematic research on chemical components and metabolites thereof entering an animal body after the traditional Chinese medicine is taken orally, and finally identifies more than two hundred compounds from in-vivo metabolites by combining detection and identification means such as gas phase, liquid phase, mass spectrum, nuclear magnetic resonance and the like, wherein one compound is named as: 2-hexyl-5-ethyl-furan-3-sulfonic acid, formula: c₁₂H₂₀O₄S, molecular weight: 260.35, the structural formula is as follows:

then, the applicant conducts high-throughput activity screening on the two hundred or more compounds, and discovers a large number of ingredients with pharmacological activity, wherein a lot of activities are related to the pharmacological action of the Xiaojin capsules, and the ingredients are inferred to be the pharmacodynamically active ingredients of the Xiaojin capsules. The compound of the invention is one of the components.

Disclosure of Invention

The invention aims to provide a new application of a compound in preparing anti-mycobacterium tuberculosis drugs.

The applicant firstly verifies the inhibition effect of the related compound on mycobacterium tuberculosis through an in vitro bacteriostasis test. And then, establishing a guinea pig lymphoid tuberculosis animal model by subcutaneously injecting mycobacterium tuberculosis bacteria, and investigating the influence of the compound on the number and weight of the swollen lymph nodes of the model animal, and finding that the number and the weight of the swollen lymph nodes of low, medium and high dose test groups are reduced and are dose-dependent, which indicates that the compound also has a better effect in vivo.

Therefore, the compound can be used for preparing anti-mycobacterium tuberculosis medicaments or preparing medicaments for treating diseases caused by mycobacterium tuberculosis, including lymphoid tuberculosis.

In the preparation of the medicament, the compound related to the invention can be used as an active ingredient alone or in combination with other antitubercular medicaments such as rifampicin, isoniazid, p-aminosalicylic acid, ethambutol and the like. Of course, when the medicines are prepared, pharmaceutically acceptable carriers can be added to prepare a preparation form suitable for clinical use.

Detailed Description

The present invention will now be described in further detail with reference to specific examples, which are provided for illustration of the present invention and are not intended to limit the scope of the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified, and materials, reagents and the like used therein are commercially available without otherwise specified.

1 Material

1.1 animals

Guinea pig, male, weighing 290-330 g, was provided by the laboratory animal center of Tongji medical college, university of science and technology, Huazhong.

1.2 strains

Mycobacterium tuberculosis Standard strain H37Rv, supplied by the provincial center of Hubei province.

1.3 drugs

2-hexyl-5-ethyl-furan-3-sulfonic acid, isolated from navy military medical university, with a purity of > 98%.

2 method

2.1 in vitro anti-tubercle bacillus action

The standard strain of mycobacterium tuberculosis is diluted to 3-6 multiplied by 10 by sterile normal saline¹⁰A/mL bacterial solution was used for the culture experiment. Preparing a drug-containing culture medium: the drug stock solution was diluted to low, medium, and high concentrations (40, 80, 160. mu.g/mL) with Roche medium, the positive control rifampicin was diluted to 5. mu.g/mL with Roche medium, and 0.1% DMSO was added to the blank medium as a negative control. All the culture media containing the medicines are sterilized by steam for 1 hour and cooled for standby. 5mL of each drug-containing culture medium is put in a culture test tube, the bacteria liquid is evenly inoculated on each culture medium inclined plane at the ratio of 1:100, the culture is carried out at the temperature of 37 ℃, the observation is carried out after 2 weeks, and the bacterial colony in the culture mediumA number of 5 or more, as "-"; the number of colonies in the culture medium is 2-4, and the colonies are marked as "+"; the number of colonies in the medium was 1 and below and was marked as "+".

2.2 action on treating scrofula

The compound with low, medium and high dose and the positive control drug are respectively diluted to 6, 12 and 24mg/mL suspension by 0.5 percent of CMC-Na to be used as gastric lavage fluid. The guinea pig is injected with 0.1mg/mL of Mycobacterium tuberculosis standard strain subcutaneously before sternum, tuberculin test is performed after one week, 30 guinea pigs with positive tuberculin reaction are taken and divided into 5 groups at random, and 6 guinea pigs in each group. The low, medium and high dose test groups respectively comprise 30mg/kg, 60mg/kg and 120mg/kg of gastric lavage compounds; the positive control group is infused with 30mg/kg of rifampicin; negative control group was intragastrically administered with 0.5% CMC-Na. The gavage administration was started the second week after the infection with tubercle bacillus, and after 2 months of continuous administration, weighing was performed, 20% pentobarbital was intraperitoneally injected, the number of swollen lymph nodes in the neck and axilla was dissected and observed, and the lymph nodes were taken and weighed.

3 results

2.1 in vitro anti-tubercle bacillus action of Compounds

The activity of the compound against mycobacterium tuberculosis is investigated by culturing mycobacterium tuberculosis in a drug-containing culture medium, and the results are shown in table 1. The results show that the positive control group, the medium and high dose test groups show obvious anti-mycobacterium tuberculosis activity, and the low dose test group also shows more obvious anti-mycobacterium tuberculosis activity.

In vitro anti-tubercle bacillus activity of the compounds of Table 1

Grouping	Concentration of	Activity against tubercle bacillus
			Negative ofControl group	0	-
Positive control group	5μg/mL	++
			Low dose group	40μg/mL	+
Middle dose group	80μg/mL	++
			High dose group	160μg/mL	++

2.2 action on treating scrofula

The number of lymph nodes with enlarged neck and axilla and the weight results of each test group are shown in Table 2. The result shows that compared with a negative control group, the number of swollen lymph nodes and the weight of lymph nodes of the animals in the low, medium and high dose test groups are reduced, and show a dose-dependent relationship, and compared with a positive control group, the result of the high dose test group has no statistical difference with the result, and the compound has a better effect of treating the lymphoid tuberculosis.

TABLE 2 therapeutic Effect of Compounds on lymph node

Grouping	Dosage form	Number of lymph nodes with swelling	Lymph node weight (g)
				Negative control group	0	4.8±0.8	0.60±0.12
Positive control group	30mg/kg	0.5±0.5^**	0.22±0.05^**
				Low dose group	30mg/kg	2.2±0.8^**	0.50±0.10
Middle dose group	60mg/kg	1.0±0.6^**	0.39±0.08^*
				High dose group	120mg/kg	0.5±0.5^**	0.24±0.06^**

^*P<0.05, relative to negative pairAccording to the group, the difference has statistical significance;^**P<0.01, there was a significant difference relative to the negative control group.

4 conclusion

Research results show that the compound shows obvious anti-tubercle bacillus activity in vivo and in vitro, and can be used for treating diseases such as lymphoid tuberculosis caused by mycobacterium tuberculosis.

Attached: raw data

。

Claims

1. The application of the compound with the structure shown as the following formula in preparing the anti-mycobacterium tuberculosis medicament,

2. use of a compound as claimed in claim 1 for the preparation of a medicament for the treatment of lymphoid tuberculosis.