CN112569232B - Pharmaceutical composition for inhibiting tuberculosis bacillus - Google Patents

Abstract

The invention relates to a pharmaceutical composition for inhibiting tubercle bacillus, which comprises kantazoxamine, bedaquiline, pretomanid and a pharmaceutically acceptable carrier. The invention discloses that the kantazodone (MRX-I) has excellent activity of inhibiting tubercle bacillus for the first time, and the combination medication scheme of the kantazodone (MRX-I) and the bedaquiline and Pretomanid shows excellent antibacterial activity.

Description

Pharmaceutical composition for inhibiting tubercle bacillus

Technical Field

The invention provides a pharmaceutical composition for inhibiting tubercle bacillus, and belongs to the field of medicines.

Background

Tuberculosis is a chronic infectious disease caused by tubercle bacillus, and is one of the major public health problems of global attention at present. The disease was declared by the World Health Organization (WHO) in 1990 as a global epidemic disease, which has affected more than one third of the world population. In particular, multi-drug resistant tuberculosis (MDR-TB) and widely drug resistant tuberculosis (XDR-TB) have appeared since the 80 s of the 20 th century, and the situation of tuberculosis prevention and control has become increasingly severe. The multi-drug resistant tuberculosis refers to tubercle bacillus resistant to any two or more drugs of streptomycin, isoniazid, rifampicin, ethambutol and pyrazinamide. The widely drug-resistant tuberculosis is a tubercle bacillus which is resistant to at least one of any fluoroquinolone drugs and three two-line injection drugs (patulin sulfate, kanamycin and amikacin) except for the multi-drug-resistant tuberculosis.

In order to cope with the urgent situation of rapid diffusion of drug-resistant tuberculosis, the world health organization proposes a multi-level combined drug administration scheme based on the current clinical first-line and second-line nuclear resistant drugs, and the occurrence probability of drug-resistant bacteria is reduced by avoiding drug cross drug resistance through the combined treatment of multiple mechanism drugs. However, in the long term, there are still two problems with the current control of tuberculosis. First, there is a lack of drugs that cure rapidly. The existing drug use scheme needs uninterrupted treatment for more than 6 months, has high treatment cost and great management difficulty, and is very easy to cause drug-resistant tuberculosis once patients are separated from the treatment. Secondly, the problem of drug resistance is becoming serious, especially for multi-drug resistant and widely drug resistant tuberculosis, there is currently a lack of effective treatment means, which poses a serious threat to the control of tuberculosis. Therefore, the development of a new generation of drugs which can treat drug-resistant tubercle bacillus infection and can quickly take effect has become urgent matters for the prevention and treatment of the modern tuberculosis.

Linezolid (Linezolid) is the earliest approved oxazolidinone antibacterial agent on the market and plays a great role in the treatment of multi-drug resistant tuberculosis (MDR-TB) and broad drug resistant tuberculosis (XDR-TB) infection, but the safety and tolerance of Linezolid for long-term use are still worth focusing, which limits the upper limit of administration of Linezolid and makes it difficult to treat drug resistant tubercle bacillus infection rapidly and effectively. In addition, the previous clinical study found that the proportion of linezolid that was discontinued due to severe adverse reactions was 15.81%.

Bedaquinine (Bedaquinine or TMC 207), produced by Johnson pharmaceutical Co., ltd. In the United states, approved for sale by the United states FDA on day 28 of 2012, was the first new antitubercular drug with a novel mechanism of action since Rifampicin (RFP) in 1974. Is also the first new drug in the world to be approved by the FDA since the 70 s of the 20 th century for the treatment of multi-drug resistant tuberculosis. Bedapsone may cause side effects of QT prolongation.

The us FDA at 2019, 8, announced that Pretomanid developed by the non-profit organization global tuberculosis drug development consortium (TB Alliance) was approved for the treatment of specific highly resistant Tuberculosis (TB) patients in combination with bedaquiline (bedaquiline) and linezolid. This is the third new anti-tuberculosis drug approved by the FDA for the last 40 years, and is the first new tuberculosis drug developed by non-profit organizations and marketed.

Applicant's prior chinese patent application CN101720325A discloses an oxazolidinone antibacterial compound, wherein the compound kantazoxamine (Contezolid, MRX-I) is disclosed, which shows high antibacterial activity in both in vivo and in vitro antibacterial models, but no related report on the study of anti-tubercle bacillus combination therapy is currently seen.

Disclosure of Invention

In view of the above problems, the present invention aims to provide a safe, highly resistant and fast acting compound and pharmaceutical composition for the treatment of drug-resistant tubercle bacillus infections.

The invention provides a pharmaceutical composition for inhibiting tubercle bacillus, which comprises the following components:

a compound of formula I or a pharmaceutically acceptable salt thereof, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable carrier;

The compound (kantazosin, MRX-I) in the three-medicine combined composition provided by the invention has equivalent antitubercular activity with linezolid, and has toxicity obviously lower than linezolid in long-term toxicity evaluation experiments of rats and dogs, and better clinical use safety, tolerance and compliance compared with linezolid. The inhibition effect of the composition containing the three-medicine combination provided by the invention on the tubercle bacillus is superior to that of the combination of the bedaquiline/Pretomanid.

Preferably, the mass ratio of the compound of formula II and the compound of formula III to the compound of formula I or the pharmaceutically acceptable salt thereof is (0.3-2.4): 1.

More preferably, the mass ratio of the compound of formula II and the compound of formula III to the compound of formula I or the pharmaceutically acceptable salt thereof is (0.5-1.0): 1.

In a preferred example, the mass sum of the compounds of formula II and III to the mass ratio of the compound of formula I or a pharmaceutically acceptable salt thereof is 5:8.

Preferably, the mass ratio of the compound of the formula II to the compound of the formula III is 1 (0.5-8).

In a preferred example, the mass ratio of the compound of formula II to the compound of formula III is 1:4.

The present invention provides for the first time a combination regimen of conazolamine (MRX-I) with bedaquiline and Pretomanid. The combined medication scheme is superior to the existing combination of the bedaquiline/Pretomanid, and is expected to become a new generation of medicine which has quick effect and can treat multi-drug resistant tuberculosis and wide drug resistant tuberculosis infection.

Detailed Description

The invention will be further described in connection with the following embodiments, it being understood that the following embodiments are only illustrative of the invention and not limiting thereof.

Firstly, the invention provides an application of a compound of formula I or pharmaceutically acceptable salt thereof in preparing a drug for inhibiting tubercle bacillus,

The preparation of the compound of formula I (Kangtai Azolamide, MRX-I) can be found in the applicant's previous China patent application CN101720325A.

Pharmaceutically acceptable salts of the compounds of formula I include, but are not limited to, (1) salts of the compounds of formula I with inorganic or organic acids to form acid addition salts, and (2) salts of the compounds of formula I in which the acidic proton is replaced by a metal ion or coordinated to an organic base. Such as hydrochloric acid, hydrobromic acid, phosphoric acid, etc., such as acetic acid, lactic acid, tartaric acid, citric acid, salicylic acid, etc., such as sodium ion, potassium ion, calcium ion, etc., such as ethanolamine, N-methyl glucamine, etc.

In vitro antitubercular Activity test of Compounds of formula I

MIC (minimum inhibitory concentration) was determined using a microplate, and the compounds of formula I were tested for their inhibitory activity against clinically isolated resistant strains FQ-R1, INH-R2, RIF-R1, RIF-R2 (strains supplied by IDRI company) against the existing antitubercular drugs rifampicin, isoniazid, levofloxacin, linezolid. Test methods Compounds were dissolved in DMSO and serially diluted 2-fold into 96-well plates containing 7H9-Tw-OADC medium at 20 different concentrations with a final DMSO solubility of 2%. The highest drug concentration was 200uM. Each plate contained a vehicle control, 100uM rifampicin control. Various tubercles were added to the 96-well plate and cultured for 5 days, and bacterial growth was detected by OD ₅₉₀. MIC values (minimum concentration at which growth was completely inhibited) were obtained by calculating a dose response curve (using GRAPHPAD PRISM to model golmpertz's model). The results are shown in Table 1 below.

Table 1:

As can be seen from Table 1, the compound of formula I in the invention has an anti-tuberculosis activity equivalent to linezolid, and has no cross drug resistance with rifampicin, isoniazid and levofloxacin, namely, has good antibacterial activity on drug resistant strains of rifampicin, isoniazid and levofloxacin, can treat tubercle bacillus infection, has the advantage of reducing the generation of drug resistant bacteria, and has the potential of clinical combined administration.

The compound of formula I was subjected to a toxicity evaluation test for 13 weeks continuous administration in SD rats, and after administration by gavage at different doses, the physiological and laboratory test indicators of the rats were observed to confirm the maximum dose (NOAEL) at which no adverse reaction was observed. Experimental methods the compound of formula I is dissolved in a 0.5% sodium hydroxymethyl cellulose solution and the formulation is administered to the animal by oral gavage twice daily (once each in the morning and twice at about 10 hours intervals) for 13 weeks, at a volume of 10mL/kg/dose, and the formulation is removed from the refrigerator at least 15 minutes prior to administration and allowed to stand at room temperature with continuous stirring until the end of administration. Animals were observed twice daily (morning and afternoon) with the primary observations being death, abnormal manifestations, pain or discomfort. Cage side observations were made once daily for toxicity test animals at the pre-dosing, dosing and recovery periods, and observed abnormal findings were recorded. Wherein during the dosing period, a cage side observation was performed about 1 hour after the morning dosing (based on the last animal/group dosing end time), and the recovery period observation time was close to the dosing period. Each toxicity test animal was subjected to a detailed clinical observation prior to pre-dosing grouping, once a week prior to dosing on day 1 of the dosing period, then 1 time a week to week 13, once on the day of sacrifice at the end of the dosing period (limited to animals sacrificed at the end), once on day 1 of recovery period, then once a week to week 4, and once on the day of death at the end of the recovery period. On day 42 of dosing (all surviving animals), day 92 (animals were sacrificed only at the end), day 29 of recovery (animals were sacrificed only at the end of recovery), blood and urine samples were collected once each. Performing hematology detection, coagulation detection, blood biochemistry detection, urine routine detection and bone marrow smear. In the test, NOAEL of the compound of the formula I reaches 100mg/kg, and the literature reports that linezolid is only 10mg/kg in the same test, which proves that the compound of the formula I has better safety than linezolid and is more suitable for the requirement of tuberculosis infection requiring long-term treatment.

In a second aspect, the present invention provides a pharmaceutical composition for inhibiting tubercle bacillus comprising a compound of formula I or a pharmaceutically acceptable salt thereof, a compound of formula II (bedaquiline), a compound of formula III (formula Pretomanid), and a pharmaceutically acceptable carrier;

in the mouse-combined infection model, a comparative test of bedaquiline/Pretomanid, bedaquiline/Pretomanid/linezolid, and bedaquiline/Pretomanid/compound of formula I was performed at different doses for 4 weeks, and the results are shown in table 2.

TABLE 2

Test method female BALB/c mice were infected with approximately 4log10 CFU of H37Rv tubercle bacillus by aerosol and after 2 weeks of feeding, started to receive lavage drug treatment. Control mice were sacrificed on the first day after infection and prior to initiation of drug treatment, lungs were dissected, and the amount of bacterial infection after infection and before treatment was determined. Bedazomet is formulated every two weeks in an acidic HPCD solution, pretomanid in a CM-2 solution, linezolid in a 0.5% methylcellulose solution, and the compound of formula I in a 0.5% sodium hydroxymethylcellulose solution. As shown in Table 2, the drugs of the compounds of formula I (Mrx) 50mg/kg,100mg/kg and 200mg/kgmg/kg were administered in combination with the drugs of the bedaquiline (B) 25mg/kg, pretomanid (Pa) mg/kg, linezolid (L) 50mg/kg and 100mg/kg, respectively (L50 means that 50mg/kg of linezolid was administered to the mice to be tested); L100 refers to administration of 100mg/kg linezolid to a subject mouse; mrX50 means MRX-I of 50mg/kg to the test mice, mrX100 means MRX-I of 100mg/kg to the test mice, mrX200 means MRX-I of 200mg/kg to the test mice, BPa means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg) to the test mice, BPaL means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg)/linezolid (50 mg/kg) to the test mice, BPaL means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg)/linezolid (100 mg/kg) to the test mice, BPaMrx means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg)/MRX-I (50 mg/kg) to the test mice, BPaMrx means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg)/X-56I) to the test mice, BPaL means Beda quinoline (25 mg/kg)/Pretomanid (100 mg/kg)/linezolid (100 mg/kg) to the test mice), BPaMrx means Beda quinoline (25 mg/kg)/Pretomanid (50 mg/kg) to the test mice), and the Beda quinoline (25 mg/kg)/linezolid (200 mg/kg) to the test mice. The stomach was irrigated once a day for 5 consecutive days a week. after 6 weeks of continuous administration, mice were dissected and the amount of bacterial infection in the lungs was determined. As can be seen from table 2, the drug combination dosing regimen (BPaMrx) provided by the present invention was comparable to bedaquiline/Pretomanid/linezolid (BPaL), and superior to bedaquiline/Pretomanid (BPa). Moreover, as the toxicity of the compound shown in the formula I is lower than that of linezolid, the tolerance is high, the administration dosage can be increased, for example, the lung infection of a tested mouse can be cleared after BPaMrx and 200 administration for six weeks, and the rapid treatment effect is achieved.

The above-mentioned medicine combination only shows the preferred proportion of every component, and its dosage ratio can be properly regulated. For example, the total amount of bedaquiline and Pretomanid and the compound of formula I is preferably (0.3-2.4): 1, more preferably (0.5-1): 1, and one preferred example is 5:8. The ratio of the bedaquiline to Pretomanid may be 1 (0.5 to 8), preferably 1:4.

The pharmaceutical composition may be an injectable, oral, inhalable or topical composition. For example, together with carriers well known in the art, such as excipients, for example, in the form of tablets, pills, capsules, suppositories, sterile injectable solutions or sterile powders and the like. It will be appreciated that the pharmaceutical compositions may be administered simultaneously or may be administered separately or in combination.

The various aspects described above are a brief description of the present invention and one of ordinary skill in the art, after reading the foregoing specification, will be able to make modifications, equivalents, and other types of modifications to the invention set forth herein. Accordingly, the description should be regarded as illustrative.

Claims (1)

1. A pharmaceutical composition for inhibiting Mycobacterium tuberculosis, characterized in that the pharmaceutical composition comprises: A compound of formula I or a pharmaceutically acceptable salt thereof, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable carrier; in, The mass ratio of the compound of formula II and the compound of formula III to the compound of formula I or a pharmaceutically acceptable salt thereof is 5:8; the mass ratio of the compound of formula II to the compound of formula III is 1:4.