A pharmaceutical composition comprising clarithromycin and sudapyridine

Abstract

The invention belongs to the field of medical technology, and specifically relates to a pharmaceutical composition comprising clarithromycin and sudapyridine. The technical problem solved is that the drug selection for inhibiting mycobacterium abscessus is limited, the treatment cycle is long, the disease is prone to recurrence, and the drug resistance to commonly used antibiotics in clinical practice is high. The pharmaceutical composition inhibits mycobacterium abscessus by combining clarithromycin and sudapyridine, wherein sudapyridine has good activity in inhibiting mycobacterium abscessus, clarithromycin has good safety and good anti-mycobacterium abscessus activity, the pharmaceutical composition can increase the blood concentration of sudapyridine in tissue within 16 hours by combining clarithromycin and sudapyridine, and a drug combination scheme having better mycobacterium abscessus inhibition activity and improving the cure rate of mycobacterium abscesses is formed by combining clarithromycin and sudapyridine. The specific technical scheme is that the ratio of clarithromycin and sudapyridine is preferably 15-20:9-15.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a pharmaceutical composition comprising clarithromycin and sudapyridine. Background Art

[0002] Nontuberculous mycobacteria refer to mycobacteria other than Mycobacterium tuberculosis complex and Mycobacterium leprae. They mainly infect the lungs, causing gradual failure of lung function, and infect other organs of the body through the blood and lymph, leading to critical illness and death. The isolation rate of nontuberculous mycobacteria in my country has been increasing year by year, from only 4.9% in 1990 to 11.1% in 2000 and 22.9% in 2010, and continued to rise after 2010.

[0003] Mycobacterium abscessum (MAB) belongs to the group of fast-growing mycobacteria among non-tuberculous mycobacteria. It can invade the lungs, skin and soft tissues, bones and joints, lymph nodes and other tissues and organs of the human body and cause local or disseminated infections. After suffering from this disease, patients will have symptoms of cough and sputum. As the disease worsens, they will also have symptoms of chest pain and chest tightness. Due to the special bacterial structure and special drug resistance mechanism of MAB, it has different degrees of resistance to commonly used antibiotics in clinical practice, which makes the treatment of MAB full of difficulties, including limited drug selection, long treatment cycle, easy recurrence and easy to develop drug resistance.

[0004] MAB also has a resistance mechanism, which can make MAB resistant to macrolide drugs after long-term interaction with them, and through a series of complex mechanisms, the activity of bacterial efflux pumps changes. Patients with MAB lung infection often have a history of underlying diseases and long-term use of antibiotics, which may induce changes in the efflux of MAB to different antibiotics.

[0005] The new anti-tuberculosis drug bedaquiline kills Mycobacterium tuberculosis by inhibiting adenosine triphosphate (ATP) synthase. Previous studies have shown that it also exhibits good activity against MAB, with most minimum inhibitory concentrations (MIC) < 1μg / ml. However, bedaquiline has severe cardiotoxicity, and studies have found that MAB causes a high rate of QT prolongation, so the U.S. Food and Drug Administration (FDA) has issued a black box marketing warning.

[0006] Sudapiridine (WX-081), as a new compound, is my country's first Class 1.1 innovative drug in the field of tuberculosis. It is an oral preparation obtained by optimizing the chemical structure of bedaquiline and affects the growth of Mycobacterium tuberculosis by inhibiting ATP synthase. WX-081 applied for new drug research in 2017 and was approved by the Chinese FDA for clinical research in 2018. Animal experiments showed that it has good oral bioavailability and wide tissue distribution, and no effects on blood pressure, heart rate and electrocardiogram were observed after conversion to adult dosage. Phase I-II clinical trials showed that it has excellent pharmacokinetic characteristics, good bioavailability, and a linear relationship between exposure and dose. The maximum blood drug concentration (Cmax) at the same dose is 1.3-1.5 times that of bedaquiline. Because WX-081 has only been used for basic and clinical research on tuberculosis, there is no clinical verification for MAB, especially the research on the combination regimen containing WX-081 for the treatment of MAB, and there are no research results at home and abroad. Summary of the invention

[0007] In a first aspect, the present invention provides a pharmaceutical composition for inhibiting MAB, wherein the pharmaceutical composition contains clarithromycin and WX-081, wherein clarithromycin can increase the blood concentration of WX-081.

[0008] Furthermore, the pharmaceutical composition contains clarithromycin and WX-081 in a ratio of 10-30:5-20.

[0009] Furthermore, the pharmaceutical composition preferably contains clarithromycin and WX-081 in a ratio of 15-20:9-15.

[0010] Furthermore, the pharmaceutical composition may be a pharmaceutical preparation, which is a combination preparation containing clarithromycin and WX-081 mixed together, or may be two independent pharmaceutical preparations containing clarithromycin and WX-081 respectively.

[0011] Furthermore, the separate pharmaceutical preparations may be administered simultaneously or sequentially.

[0012] In one embodiment, the pharmaceutical composition is preferably in the form of a tablet, which is an independent formulation of clarithromycin and WX-081 and is administered to a subject in a sequential manner.

[0013] Furthermore, the pharmaceutical preparation also includes pharmaceutically acceptable excipients.

[0014] Furthermore, the excipient is a pharmaceutical excipient that enhances the efficacy of the pharmaceutical composition, reduces toxicity and / or alleviates toxic side effects.

[0015] The pharmaceutical preparation can be selected from pills, pills, decoctions, granules, capsules, powders, tablets, injection preparations and the like.

[0016] Furthermore, the pharmaceutical preparation comprises a third active ingredient, which is an active ingredient having anti-MAB activity.

[0017] Furthermore, the third active ingredient includes but is not limited to moxifloxacin, clofazimine, linezolid and sulfonamides.

[0018] In a second aspect, the present invention provides an application of a pharmaceutical composition in the preparation of a drug for inhibiting MAB.

[0019] Furthermore, the pharmaceutical composition contains clarithromycin and WX-081.

[0020] Furthermore, the pharmaceutical composition contains clarithromycin and WX-081 in a ratio of 10-30:5-20.

[0021] Furthermore, the pharmaceutical composition preferably contains clarithromycin and WX-081 in a ratio of 15-20:9-15.

[0022] Furthermore, the application is to increase blood drug concentration.

[0023] Furthermore, the blood drug concentration is the blood drug concentration of WX-081 increased by clarithromycin.

[0024] Beneficial Effects

[0025] The present invention combines clarithromycin and WX-081 to form a drug combination scheme with better anti-MAB activity and improved MAB cure rate. It has been verified that the combined use can increase the blood concentration of WX-081 and the content in the spine and lung tissues. DETAILED DESCRIPTION

[0026] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in the embodiments described below can be combined with each other as long as they do not conflict with each other.

[0027] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0028] The structural formula of "Sudapyridine" (WX-081) involved in this article is as follows:

[0029]

[0030] Example 1 Preparation of Pharmaceutical Composition Tablet I

[0031] The first step is to crush, sieve and mix clarithromycin and WX-081 evenly.

[0032] In the second step, corresponding auxiliary materials are added to the mixture obtained in the first step.

[0033] The third step is to prepare dry granules, whole granules and tablets from the mixture obtained in the second step to obtain clarithromycin-WX-081 tablets.

[0034] In the fourth step, the auxiliary materials described in the second step include: fillers, binders, disintegrants, and lubricants.

[0035] Example 2 Preparation of Pharmaceutical Composition Tablet II

[0036] The first step is to crush and sieve clarithromycin.

[0037] The second step is to crush and sieve WX-081.

[0038] In the third step, the fine powders obtained in the first and second steps are added with corresponding auxiliary materials, dried, granulated, and tableted to obtain clarithromycin tablets with a drug content of 250 mg per tablet and WX-081 tablets with a drug content of 150 mg per tablet.

[0039] In the fourth step, the auxiliary materials described in the third step include: fillers, binders, disintegrants, and lubricants.

[0040] Example 3 Study on tissue distribution of WX-081 in rats

[0041] 1. Experimental Materials

[0042] Experimental animals: 12 SD rats that passed quarantine, half male and half female, SPF grade, weighing 180-250 g, were purchased from Hangzhou Medical University.

[0043] The 12 SD rats were randomly divided into two groups according to gender and weight, half of which were male and half were female. There was no statistical difference in the average weight of the animals in each group after grouping.

[0044] Rats were housed in a 485×350×200mm 3 (length × width × height) PP cages, with no more than 5 animals per cage. Record the environmental parameters of the animal room during the breeding period. During the experiment, animals of other species shall not be kept in the same room area.

[0045] Experimental reagents: WX-081, bedaquiline, loratadine, formic acid, methanol, acetonitrile, and normal saline.

[0046] Table 1 Rat dosage design

[0047]

[0048] The drug was administered by intragastric gavage. The dosage was calculated based on the body weight on the first day (overnight fasting). The administration time was 9:00-10:00 am, once a day.

[0049] Rats were fasted for at least 12 hours before administration, and were fed uniformly 4 hours after administration, but were not allowed to drink water. Rats were killed by bleeding from the femoral artery at 3 time points after administration, and plasma was collected, and brain tissue, spine and lung tissue were immediately removed and frozen for later use. WX-081 and its combined administration groups were administered 20 minutes, 1 hour and 16 hours after administration. The collected tissue samples were placed in an ice box and stored in a -80°C refrigerator for testing.

[0050] 2. Experimental methods

[0051] 2.1 LC-MS / MS analysis conditions

[0052] Chromatographic column: Agilent XDB C18 (50 mm × 3.5 mm, 2.1 μm); flow rate: 0.3 mL / min;

[0053] Column temperature: 35°C; injection volume: 5 μL. Mobile phase A: 0.1% formic acid solution, mobile phase B: methanol, gradient elution see Table 2

[0054] Table 2 Gradient elution conditions

[0055]

[0056] 2.2 Mass spectrometry conditions

[0057] ESI source, positive ion mode, MRM detection, acquisition of each compound signal, see Table 3. The main parameters are: gas temp: 300℃; gas flow: 5L / min; nebulizer: 45psi; sheath gas temp: 400℃; sheath gas flow: 11L / min; capillary: 3.5kV; nozzle voltage: 500V.

[0058] Table 3 Detection conditions of target compound and internal standard LC-MS / MS

[0059]

[0060] 2.3 Preparation of standard and internal standard stock solutions

[0061] Accurately weigh 10 mg of WX-081 reference substance, add 1 mL of DMSO to prepare a 10 mg / mL WX-081 stock solution, and store it at a low temperature and away from light for later use.

[0062] Accurately measure 300 mL of acetonitrile and 15 μL of LLTD (100 μg / mL) into the mobile phase bottle and mix them to prepare a 5 ng / mL LLTD solution.

[0063] 2.4 Sample processing

[0064] Take about 0.1g of rat brain tissue, spine, and lung tissue, weigh accurately, and make homogenate with physiological saline at a ratio of 1:9. Take out 50μL of brain tissue and spine homogenate sample or plasma sample and put it into a 1.5mL EP tube, and then add 150μL of acetonitrile solution containing internal standard to precipitate protein. After mixing, vortex oscillation, centrifuge at 10℃ and 14,000rpm for 10min, and take the supernatant for LC-MS / MS injection analysis.

[0065] For samples diluted 5 times: Take out 10μL of the corresponding tissue homogenate sample or plasma sample and put it into a 1.5mL EP tube, add 40μL of blank matrix, vortex to mix, and then add 150μL of acetonitrile solution containing internal standard to precipitate protein. After mixing, vortex and oscillate, centrifuge at 10℃ and 14,000rpm for 10min, and take the supernatant for LC-MS / MS analysis.

[0066] Samples diluted 10 times: Take out 5 μL of the corresponding tissue homogenate sample or plasma sample into a 1.5 mL EP tube, add 45 μL of blank matrix, vortex to mix, and then add 150 μL of acetonitrile solution containing internal standard (LLTD, 5 ng / mL) to precipitate protein. After mixing, vortex, centrifuge at 10°C, 14,000 rpm for 10 minutes, and take the supernatant for LC-MS / MS injection analysis.

[0067] 2.5 Preparation of target standard curve

[0068] Accurately pipette 47.5 μL of the homogenate sample of the spine or lung tissue or the plasma sample into a 1.5 mL EP tube, and accurately add 2.5 μL of WX-081 working solution with concentrations of 1, 2, 4, 10, 16, and 20 μg / mL in parallel to prepare standard curve samples with concentrations of 50, 100, 200, 500, 800, and 1000 ng / mL, process according to the "Sample Processing" item, and inject 5 μL for analysis under the experimental conditions. Accurately pipette 47.5 μL of brain tissue homogenate sample into a 1.5 mL EP tube, and accurately add 2.5 μL of WX-081 working solution with concentrations of 0.2, 0.4, 1, 2, 4, 10, 16, 20 μg / mL in parallel to prepare standard curve samples with concentrations of 10, 20, 50, 100, 200, 500, 800, 1000 ng / mL, and process according to the "sample processing" item. Inject 5 μL under the experimental conditions for analysis. The results are obtained with the concentration of the analyte as the horizontal coordinate and the peak area of ​​the analyte as the vertical coordinate. The weighted (W=1 / X2) least squares method is used for regression calculation, and the linear regression equation obtained is the standard curve. The linear range of WX-081 in plasma, spine and lung tissue is 50-1000 ng / mL; the linear range of brain tissue is 10-1000 ng / mL, and the linear range is good within this range, as shown in Table 4.

[0069] Table 4 Standard curve and linear range of WX-081

[0070]

[0071] To ensure reliable analytical quality determination, a working curve is carried out for each analytical batch, and three quality control samples of low, medium and high concentrations are measured at the same time. The number of quality control samples is required to be greater than 5% of the total number of unknown samples. The sample concentration and quality control sample concentration are calculated based on the working curve of the day. The accuracy value of at least 67% of the quality control samples should be within the range of ±15% of the labeled value, and at least 50% of the samples at each concentration level should meet this standard. The measurement results of the quality control samples of each compound are detailed in Table 5.

[0072] Table 5 Quality control of WX-081 during analysis

[0073]

[0074]

[0075] 3. Experimental results

[0076] The results of WX-081 content in group A and group B are shown in Table 6.

[0077] The results showed that after rats were given group A and group B doses by oral gavage, WX-081 was highest in the lungs, followed by plasma and spine, and less in the brain.

[0078] Comparing the A and B administration groups, it was found that the plasma concentration of WX-081 in both A and B administration groups increased with time. The plasma concentration at 20 minutes was group B>group A; at 1 hour, group B>group A; at 16 hours, group B>group A. This indicates that group B can increase the blood concentration of WX-081.

[0079] Regarding the distribution of WX-081 in brain tissue, no WX-081 was detected in the brain tissue of group A and group B at 20 minutes and 1 hour. At 16 hours, the concentration of WX-081 in group A was higher than that in group B, but there was no statistical difference.

[0080] In the distribution of WX-081 in the spine, the amount of WX-081 in the A and B administration groups increased with time. At 20 minutes, no WX-081 was detected. At 1 hour, group A was greater than group B, and at 16 hours, group B was greater than group A. This indicates that the B administration group can increase the content of WX-081 in the spine within 16 hours.

[0081] In the distribution of WX-081 in lung tissue, the amount of WX-081 in the A and B administration groups increased with time. Among them, at 20 minutes, group B>group A, at 1 hour, group B>group A, and at 16 hours, group B>group A, indicating that the B administration group can increase the content of WX-081 in the lung tissue.

[0082] Table 6 Content of WX-081 in tissues

[0083]

[0084]

[0085] 4. Conclusion

[0086] Compared with the WX-081 single administration group, the WX-081 and clarithromycin combination group can increase the blood concentration, spine and lung tissue content of WX-081. Compared with group A, the WX-081 content in the plasma of group B at 20min and 1h was significantly increased, and the content in the lung tissue at 1h was significantly increased.

Claims

1. A pharmaceutical composition for inhibiting Mycobacterium abscessus, the pharmaceutical composition consisting of clarithromycin and sudapyridine, wherein clarithromycin can increase the blood concentration of sudapyridine.

2. The pharmaceutical composition for inhibiting Mycobacterium abscessus according to claim 1, characterized in that: The pharmaceutical composition is a pharmaceutical preparation, which is a combination preparation of clarithromycin and sudapyridine or two independent preparations.

3. The pharmaceutical preparation for inhibiting Mycobacterium abscessus according to claim 2, characterized in that: The pharmaceutical preparation comprises a pharmaceutically acceptable excipient, wherein the excipient is a pharmaceutical excipient that can enhance the efficacy, reduce toxicity and / or alleviate side effects of the pharmaceutical composition.

4. The pharmaceutical preparation for inhibiting Mycobacterium abscessus according to claim 2, characterized in that: The pharmaceutical preparation is a pill, a pill, a decoction, a granule, a capsule, a powder, a tablet or an injection preparation.

5. The pharmaceutical preparation for inhibiting Mycobacterium abscessus according to claim 4 is selected from tablets, characterized in that: The tablets are separate formulations of clarithromycin and sudapyridine and are administered to the subject in a sequential manner.

6. Use of a pharmaceutical composition in the preparation of a drug for inhibiting Mycobacterium abscessus, wherein the pharmaceutical composition consists of clarithromycin and sudapyridine.

7. Use of the pharmaceutical composition according to claim 6 in the preparation of a drug for inhibiting Mycobacterium abscessus, characterized in that: The application is to increase blood drug concentration, and the increased blood drug concentration is that clarithromycin increases the blood drug concentration of sudapyridine.

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