KR20230138735A - Pharmaceutical Anti-Tuberculosis Composition - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating tuberculosis, which has a structure different from that of conventionally known agents, and more specifically, 4,5,6,7-Tetrahydroisoxazolo[5,4-c] represented by the following formula (1): ]Relates to a pharmaceutical composition for preventing or treating tuberculosis containing pyridin-3-ol (THIP) as an active ingredient.
[Formula 1]

Description

Anti-tuberculosis pharmaceutical composition {Pharmaceutical Anti-Tuberculosis Composition}

The present invention relates to a pharmaceutical composition for preventing or treating tuberculosis, which has a different structure from conventionally known preparations.

Tuberculosis (TB) is an infectious disease caused by infection with Mycobacterium, especially Mycobacterium tuberculosis, and has the characteristic of progressing or maintaining slowly over a long period of time compared to other infectious diseases. Mycobacterium tuberculosis is a rod-shaped bacillus with a very slow growth rate. It is surrounded by a cell wall rich in fat, so it can live for a long time even in dry conditions, and it is also called an acid-fast bacterium because it can withstand strong acids and alkalis.

Worldwide, it is estimated that one-third of the total population is infected with tuberculosis bacteria, and approximately 800 million new patients occur every year. Mycobacterium tuberculosis develops after an incubation period for a certain period of time after infection, or develops acutely without an incubation period, causing complications such as lung inflammation and asthma, leading to the death of the infected person. Most infected people are simple carriers with no symptoms, and about 1/10 of them develop the disease, and if they do not receive appropriate treatment when they develop the disease, more than half of them will die. Because simple carriers have no symptoms, they can easily spread tuberculosis to others, making it difficult to prevent and treat tuberculosis. Typical symptoms include coughing with bloody sputum, chills, cold sweats, and weight loss. Since any organ in the body can be infected, it causes various symptoms depending on the infected organ.

Currently, chemotherapy is the treatment method for tuberculosis. The first-line anti-tuberculosis drug for the treatment of early-stage tuberculosis is a combination of drugs (isoniazid, rifampin, pyrazinamide, and ethambutol). Although it is a very effective tuberculosis treatment method with a tuberculosis cure rate of about 85%, it requires continuous administration for at least 6 months or more. , it causes significant side effects, is expensive, and has unclear efficacy against multidrug-resistant tuberculosis. In particular, since one mutant strain of Mycobacterium tuberculosis appears every 10 ⁶ cell divisions, at least two types of drugs, and if possible four or more types of drugs, must be used in combination. However, if resistance to one drug develops, there is no special treatment method, such as chemotherapy. This is the biggest problem.

The mortality rate due to tuberculosis has been increasing again since the 1990s, which is believed to be mainly due to the increase in infections caused by multi-drug resistant (MDR) tuberculosis bacteria and the increase in HIV (Human Immunodeficiency Virus)/TB co-infection. Currently, there are approximately 50 million multidrug-resistant tuberculosis patients worldwide, but the treatment efficiency is only about 50%. In addition, according to the revised tuberculosis treatment guidelines in 2017, tuberculosis is one of the major opportunistic diseases that occur in HIV-infected people, and in Korea, it accounts for 11-25% of opportunistic infectious diseases in HIV-infected people. The occurrence of tuberculosis in HIV patients is more important not only because of the risk of tuberculosis, but also because it can have a negative impact on HIV disease. In the case of HIV-infected people with tuberculosis, the blood concentration of HIV increases and HIV disease may progress more rapidly (Non-patent Document 1).

Therefore, the development of a new anti-tuberculosis drug that has a different mechanism of action from existing drugs and has relatively fewer side effects is a very urgent task.

Registered Patent No. 10-1344218

Tuberculosis treatment guidelines (3rd edition) 2017.

The purpose of the present invention is to provide a pharmaceutical composition with a new structure that can effectively prevent or treat tuberculosis by inhibiting the proliferation of tuberculosis bacteria.

The present invention for achieving the above-described object is a method for preventing or treating tuberculosis comprising 4,5,6,7-Tetrahydroisoxazolo[5,4-c]pyridin-3-ol (THIP) represented by the following formula (1) as an active ingredient: It relates to pharmaceutical compositions.

[Formula 1]

It was confirmed that THIP can be usefully used in the prevention or treatment of tuberculosis by significantly reducing the survival rate of Mycobacterium tuberculosis in macrophages infected with Mycobacterium tuberculosis. Macrophages are one of the cells responsible for immunity. Generally, bacterial strains are sterilized by the phagocytosis of macrophages, but Mycobacterium tuberculosis suppresses the phagocytosis within macrophages and evades the action of anti-tuberculosis drugs by hiding within the macrophages. This is the biggest reason why tuberculosis bacteria do not respond to drugs. Therefore, the anti-tuberculous activity of macrophages indicates that cells infected with Mycobacterium tuberculosis can be effectively eliminated by the immune system. In addition, it exhibits antibacterial activity not only against active tuberculosis such as pulmonary tuberculosis, biliary tuberculosis, bone tuberculosis, throat tuberculosis, lymphatic tuberculosis, pulmonary tuberculosis, breast tuberculosis, spinal tuberculosis, or multidrug-resistant tuberculosis, but also against latent tuberculosis strains in macrophages, thereby preventing latent tuberculosis. It can also be useful for prevention or treatment. In the present invention, “latent tuberculosis” refers to inactive tuberculosis, which refers to tuberculosis in a dormant state after primary tuberculosis infection or a state of Mycobacterium tuberculosis infection without the development of disease symptoms.

The pharmaceutical composition of the present invention is prepared by itself or mixed with a pharmaceutically acceptable carrier, forming agent, diluent, etc., by a method known in the pharmaceutical field, to form powder, granules, tablets, capsules, or injections. It can be manufactured and used in dosage forms such as:

The pharmaceutical composition according to the present invention can be administered in a pharmaceutically effective amount. In the present invention, the 'pharmaceutically effective amount' means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment. The level of effective dosage depends on factors including the type of patient's disease, severity, age, gender, activity of the drug, sensitivity to the drug, time of administration, route of administration and excretion rate, duration of treatment, drugs used simultaneously, and other fields of medicine. It can be determined based on known factors. Considering all of the above factors, it is important to administer an amount that can achieve maximum effect with the minimum amount without side effects, and this can be easily determined by a person skilled in the art.

The composition of the present invention can be administered alone or in combination with other anti-tuberculosis treatments. Since one mutant strain of Mycobacterium tuberculosis appears every approximately 10 ⁶ cell divisions, it is more desirable to use at least two or more types of drugs, and if possible, four or more types of drugs in combination. When administered in combination with other anti-tuberculosis treatments, it may be administered sequentially or simultaneously with conventional treatments. The anti-tuberculosis treatments administered in combination include rifampin, isoniazid, pyrazinamide, ethambutol, streptomycin, fluoroquinolone, kanamycin, and cycloserine. (Cycloserine), Prothionamide, Levofloxacin, Moxifloxacin, Ofloxacin, Rifabutin, Capeomycin and Linezolid ( It may be one or more selected from the group consisting of linezolid, but is not limited thereto.

As described above, according to the present invention, the survival rate of tuberculosis bacteria can be significantly reduced by using THIP, which has a new structure compared to conventional anti-tuberculosis drugs, as an active ingredient, and can be usefully used as a new pharmaceutical composition for preventing or treating tuberculosis.

In addition, the composition of the present invention has an inhibitory effect on the proliferation of latent tuberculosis strains internalized in macrophages, and can prevent or improve not only active tuberculosis but also latent tuberculosis, so it can be usefully used to prevent the spread of tuberculosis by simple carriers. there is.

Figure 1 is a graph showing the anti-tuberculous efficacy of THIP against Mycobacterium tuberculosis in macrophages.

The present invention will be described in more detail below with reference to the attached examples. However, these embodiments are only examples for easily explaining the content and scope of the technical idea of the present invention, and the technical scope of the present invention is not limited or changed thereby. Based on these examples, it will be obvious to those skilled in the art that various modifications and changes are possible within the scope of the technical idea of the present invention.

[Example]

Example 1: Mycobacterium tuberculosis infection of macrophages and treatment with fenofibrate

1) Collection and culture of peritoneal macrophages

8-week-old wild-type C57BL/6 mice were injected intraperitoneally with 1 mL of 3% thioglycollate, and 3 days later, they were injected with cooled PBS (phosphate) containing 3% FBS (fetal bovine serum, Gibco; 16000-044). 3 mL of buffer saline) was injected into the abdominal cavity. Abdominal lavage was collected and centrifuged at 4°C and 1,500 rpm for 8 minutes to obtain a macrophage pellet. The collected macrophages were suspended in DMEM (Lonza; 12-604F) containing 10% FBS and penicillin-streptomycin-amphotericin B (Lonzal; 17-745E) and then cultured overnight.

2) Mycobacterium tuberculosis infection and drug treatment

Pathogenic tuberculosis bacteria Mycobacterium tuberculosis (Mtb) or M. bovis Bacillus Calmette-Guerin (BCG) were cultured in the mouse abdominal macrophage medium prepared in 1). The average number of infected bacteria per cell (multiplicity of infection, MOI) was 1:1. This was added to allow infection for 2 hours. After washing the extracellular bacteria with PBS, they were treated with THIP at a concentration of 10 or 50 μM and cultured for 3 days. As a control, 0.1% DMSO was used.

Example 2: Analysis of the impact of THIP on Mycobacterium tuberculosis survival rate

For the Mycobacterium tuberculosis-infected macrophages treated with fenofibrate according to 2) of Example 1, the culture medium of the macrophages was removed immediately before, 3 days after, or 7 days after treatment with fenofibrate, and then sterilized distilled water was added to burst the macrophages. To identify tuberculosis bacteria present in macrophages, 7H10 medium was used, and after culturing in an incubator at 37°C for about 14 days, colony forming units (CFU) were measured to confirm intracellular survival (ICS).

Figure 1 shows the results, and it was confirmed that the intracellular survival rate of Mycobacterium tuberculosis and BCG decreased significantly in a concentration-dependent manner 3 days after addition of THIP.

Claims (5)

A pharmaceutical composition for preventing or treating tuberculosis comprising a compound of the following formula (1) as an active ingredient.
[Formula 1]
In claim 1,
The tuberculosis is pulmonary tuberculosis, biliary tuberculosis, bone tuberculosis, throat tuberculosis, lymphatic tuberculosis, lung tuberculosis, breast tuberculosis, spinal tuberculosis, or multidrug-resistant tuberculosis. A pharmaceutical composition for preventing or treating tuberculosis.
In claim 1,
A pharmaceutical composition for preventing or treating tuberculosis, characterized in that it exhibits antibacterial activity against tuberculosis strains latent in macrophages.
In claim 1,
A pharmaceutical composition for preventing or treating tuberculosis, characterized in that it is administered in combination with other anti-tuberculosis treatments.
In claim 4,
The anti-tuberculosis treatments include rifampin, isoniazid, pyrazinamide, ethambutol, streptomycin, fluoroquinolone, kanamycin, and cycloserine. , Prothionamide, Levofloxacin, Moxifloxacin, Ofloxacin, Rifabutin, Capeomycin, and Linezolid. A pharmaceutical composition for preventing or treating tuberculosis, characterized in that it contains at least one selected from the group consisting of.