CN1857220B - A sustained-release agent for anti-tuberculosis drugs - Google Patents

Abstract

The slow released antituberculotic preparation is implanting agent or injection with slow releasing of medicine in the local tuberculosis focus to maintain the local effective medicine concentration while lowing the systemic toxicity. The slow released injection consists of slow released microsphere and solvent. The slow released microsphere consists of slow releasing supplementary material and antituberculotic selected from rifampicin, rifamycin, rifapentine and/or rifabutine. The solvent is special solvent containing suspending agent of carboxymethyl cellulose sodium, etc. and has viscosity of 100-3000 cp at 20-30 deg.c. The slow releasing supplementary material is selected from EVAc, PLA, PLGA, etc. The slow released preparation may be prepared with slow released microsphere. The present invention has obvious and unique treating effect on various kinds of intractable tuberculosis.

Description

A sustained-release agent for anti-tuberculosis drugs

(1) Technical field

The invention relates to an anti-tuberculosis drug sustained-release agent, belonging to the technical field of drugs. Specifically, the invention provides a slow-release preparation containing anti-tuberculosis drugs, mainly slow-release injections and slow-release implants. The slow-release agent is mainly applied locally, and can obtain and maintain an effective drug concentration in the local area of the tuberculosis lesion.

(2) Background technology

Tuberculosis, represented by pulmonary tuberculosis, is a disease that seriously affects people's health. It is widespread all over the world and has killed hundreds of millions of people. People call it the white plague. There was a saying that "ten tuberculosis and nine deaths". With the advent of anti-tuberculosis drugs such as streptomycin, tuberculosis became a treatable disease. However, with people's ignorance of its severity, obvious insufficient investment in related prevention and treatment, and irregular treatment, the resulting drug resistance of tuberculosis has become the most difficult problem in the successful treatment of the disease.

Because it is very contagious, its incidence rate is increasing at a rate of 1% in the world every year in recent years. Among the three diseases identified by the World Health Organization that need to be controlled globally, tuberculosis is second only to AIDS and ahead of malaria. One person is infected with tuberculosis every second in the world, and 3 to 4 people die from tuberculosis every minute. At present, there are still about 3 million people dying from tuberculosis every year in the world on average. The current epidemic of tuberculosis in China is very serious, and it has become one of the 22 countries with a high burden of tuberculosis in the world. At present, there are about 6 million tuberculosis patients in China, accounting for one-third of the global number of patients, ranking second in the world. in India; drug-resistant patients in China accounted for a quarter of the world. There are at least 1.5 million new patients every year in the country, among which more than 650,000 are infectious patients. The number of deaths due to tuberculosis is as high as 250,000 each year, and most of them are young and middle-aged. It has become the number one killer as a single infectious disease. Tuberculosis requires more than six months of continuous medication to achieve a satisfactory prognosis. Due to the long treatment time, many patients may forget to take the medicine in a timely and quantitative manner, which often leads to the emergence of drug resistance. On the one hand, the treatment of drug-resistant tuberculosis patients is protracted, and on the other hand, the application of multiple drugs combined with chemotherapy is expensive. For example, more than 20,000 to 30,000 yuan are required in China. Therefore, the research and development of new and effective preparations or methods for preventing and treating tuberculosis has become an urgent worldwide issue.

At present, many new anti-tuberculosis drugs have shown good efficacy, but the effect on multidrug-resistant tuberculosis (MDR-TB) is not ideal. Because for many chronic lesions, especially local lesions, it is difficult to obtain an effective bactericidal concentration with the administration of conventional therapy. There will be many side effects if the dose is increased or the drug is taken for a long time.

(3) Contents of the invention

The present invention aims at the deficiencies in the prior art, and provides a slow-release agent containing anti-tuberculosis drugs, specifically, a slow-release agent for anti-tuberculosis drugs containing rifampicin, mainly slow-release injections and slow-release implants agent.

As a commonly used anti-tuberculosis drug, rifampicin is mainly an oral preparation in foreign countries. Even ordinary injections are not ideal, because effective drug concentrations cannot be obtained at the lesion site. Moreover, the obvious systemic toxicity and the generation of drug resistance during application greatly limit the application of this drug.

The present invention finds that making anti-tuberculosis drugs into slow-release preparations (mainly slow-release injections and slow-release implants) can not only greatly increase the local drug concentration, reduce the concentration of the drug in the circulatory system, and reduce the effect of the drug on normal tissues. It can also greatly facilitate the application of drugs, reduce the course of treatment, shorten the treatment time, reduce the complications of drugs, reduce the cost of patients, and reduce drug tolerance. The above unexpected findings constitute the main content of the present invention.

One form of the drug sustained-release preparation of the present invention is a sustained-release injection, which consists of sustained-release microspheres and a vehicle. Specifically, the sustained-release injection consists of the following ingredients:

(a) Slow-release microparticles, consisting of the following components by weight percentage:

Anti-tuberculosis drugs 1-70%

Sustained release excipients 30-99%

Suspending agent 0.0-30%

The above is weight percentage

and

(b) The vehicle is an ordinary vehicle or a special vehicle containing a suspending agent.

in,

Anti-tuberculosis drugs are selected from rifampicin and its analogues; sustained-release excipients are selected from polyphenylene, difatty acid and sebacic acid copolymer (PFAD-SA), poly (erucic acid dimer-sebacic acid) [ P(EAD-SA)], poly(fumaric-sebacic acid) [P(FA-SA)], ethylene vinyl acetate copolymer (EVAc), polylactic acid (PLA), polyglycolic acid, and glycolic acid One or a combination of copolymer (PLGA), xylitol, oligosaccharides, chondroitin, chitin, hyaluronic acid, collagen, gelatin and albumin glue; suspending agent is selected from sodium carboxymethylcellulose, (iodo)glycerin, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surface active substances, one of TWen 20, TWen 40 and TWen 80 or a combination thereof.

The analogs of rifampin include rifamycin (rifamycin), rifapentine (rifapentine, DL473, cyclopentiperazine rifamycin), the use of butin (spirocyclic piperidine rifamycin, ansamycin , LM427). The anti-tuberculosis drug of the present invention can be selected from one or more of rifampicin and its analogues.

The proportion of the above-mentioned anti-tuberculosis drugs in the composition depends on specific conditions, and may be 1%-70%, preferably 2%-50%, and most optimally 5%-40%.

The weight percentages of active ingredients and slow-release auxiliary materials in the anti-tuberculosis sustained-release microspheres of the present invention are preferably as follows:

Anti-tuberculosis drugs 2-50%

Sustained release excipients 50-98%

Suspending agent 0.0-30%

Sustained release excipients are selected from polyphenylpropane, difatty acid and sebacic acid copolymer (PFAD-SA), poly(erucic acid dimer-sebacic acid), poly(fumaric acid-sebacic acid), ethylene acetic acid Vinyl ester copolymers, polylactic acid, copolymers of polyglycolic and glycolic acids, hyaluronic acid, collagen, or gelatin.

The most preferred active ingredients and weight percentages in the sustained-release microspheres of the present invention are as follows:

(a) 5-40% rifampicin, rifamycin, rifapentine or rifabutin;

(b) 5-40% rifampicin in combination with 5-40% rifamycin, rifapentine or rifabutin;

(c) 5-40% rifamycin in combination with 5-40% rifapentine or utilibutin: or

(d) Combination of 5-40% rifapentine and 5-40% urbutin.

The most preferred percentage by weight of the slow-release auxiliary material in the slow-release microspheres of the present invention is as follows:

(1) 55-90% PLA;

(2) 50-90% PLGA;

(3) 50-85% polyphenylene;

(4) 55-90% difatty acid and sebacic acid copolymer; or

(5) 55-90% EVAc.

The above slow-release microspheres are mixed with sodium carboxymethylcellulose, (iodo)glycerin, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surface active substances, soil temperature 20, soil temperature 40 or soil temperature 80 Suspension vehicles together to make sustained-release injections. Wherein the concentration of sodium carboxymethyl cellulose in solvent can be 0.1-5%, but is preferred with 0.53%, is most preferred with 1-2%.

The peak molecular weight of polylactic acid can be, but not limited to, 5000-100,000, but preferably 20,000-60,000, most preferably 30,000-50,000; the molecular weight of polyglycolic acid can be, but not limited to, 5000-100,000, but in 20,000-60,000 is preferred, and 30,000-50,000 is the most preferred; the above polyhydroxy acids can be single or multiple. When selected alone, polylactic acid (PLA) or a copolymer of hydroxycarboxylic acid and glycolic acid (PLGA) is preferred, and the molecular weight of the copolymer can be, but not limited to, 5000-100,000, but preferably 20,000-60,000, 30,000-50,000 is the most preferred; when multiple options are selected, polymers or composite polymers or copolymers composed of different polymers are preferred, and composites containing polylactic acid or sebacic acid with different molecular weights are preferred. Polymers or copolymers are most preferred, such as, but not limited to, polylactic acid with a molecular weight of 1000 to 30000 mixed with polylactic acid with a molecular weight of 20000 to 50000, polylactic acid with a molecular weight of 10000 to 30000 and polylactic acid with a molecular weight of 30000 to 80000 PLGA is mixed, polylactic acid with a molecular weight of 20,000 to 30,000 is mixed with sebacic acid, and PLGA with a molecular weight of 30,000 to 80,000 is mixed with sebacic acid.

Among various high molecular polymers, polylactic acid, sebacic acid, mixtures or copolymers of high molecular weight polymers containing polylactic acid or sebacic acid are preferred, mixtures and copolymers can be selected from, but not limited to, PLA, PLGA, mixtures of glycolic acid and hydroxycarboxylic acids, mixtures or copolymers of sebacic acid and aromatic polyanhydrides or aliphatic polyanhydrides. The blending ratio of glycolic acid and hydroxycarboxylic acid is 10/90-90/10 by weight, preferably 25/75-75/25 by weight. The method of blending is arbitrary. The contents of glycolic acid and hydroxycarboxylic acid in copolymerization are respectively 10-90% and 90-10% by weight. Representatives of aromatic polyanhydrides are polyphenylpropanol [poly(1,3-bis(p-carboxyphenoxy)propane-sebacic acid) (p(CPP-SA)), difatty acid-sebacic acid copolymer ( PFAD-SA)], poly(erucic acid dimer-sebacic acid) [P(EAD-SA)] and poly(fumaric acid-sebacic acid) [P(FA-SA)], etc. The contents of p-carboxyphenoxypropane (p-CPP) and sebacic acid copolymerization are respectively 10-60% and 20-90% by weight, and the blending weight ratio is 10-40: 50-90, preferably by weight Ratio 15-30:65-85.

In addition to the above-mentioned excipients, other substances can also be used, see US Patent (Patent No. 4757128; 4857311; 4888176; 4789724) and "Compendium of Pharmaceutical Excipients" (page 123, published by Sichuan Science and Technology Press in 1993, edited by Luo Mingsheng and Gao Tianhui ) has been described in detail. In addition, Chinese patents (application Nos. 96115937.5; 91109723.6; 9710703.3; 01803562.0) and U.S. invention patents (patent No. 5,651,986) also list certain pharmaceutical excipients, including fillers, solubilizers, absorption accelerators, film-forming agents, gelling agents, etc. agent, preparation (or) porogen, excipient or blocker, etc.

In order to adjust the drug release rate or change other characteristics of the present invention, the monomer composition or molecular weight of the polymer can be changed, the composition and proportion of pharmaceutical excipients can be added or adjusted, and water-soluble low-molecular compounds can be added, such as, but not limited to, each sugar or salt etc. Wherein the sugar can be, but not limited to, xylitol, oligosaccharide, (sulfate) chondroitin and chitin, etc., wherein the salt can be, but not limited to, potassium salt and sodium salt, etc.

In sustained-release injections, the drug sustained-release system can be made into microspheres, submicrospheres, microemulsions, nanospheres, granules or spherical pellets, and then mixed with injection vehicles to make injections for use. Suspension-type sustained-release injections are preferred among various sustained-release injections. Suspension-type sustained-release injections are preparations obtained by suspending drug sustained-release systems containing anti-tuberculosis components in injections. The auxiliary materials used are the above-mentioned sustained-release injections. One or a combination of excipients, the solvent used is a common solvent or a special solvent containing a suspending agent. Common solvents are, but not limited to, distilled water, water for injection, physiological flushing solution, absolute ethanol or buffers prepared with various salts. The purpose of the suspending agent is to effectively suspend the drug-containing microspheres, thereby facilitating injection.

The suspending agent is selected from one of sodium carboxymethylcellulose, (iodo)glycerin, simethicone, propylene glycol, carbomer, mannitol, sorbitol, surface active substances, earth temperature 20, earth temperature 40 and earth temperature 80 or a combination thereof.

The content of the suspending agent in the common solvent depends on its characteristics, and can be 0.1-30% depending on the specific circumstances. The composition of preferred suspending agent is:

A) 0.5-5% sodium carboxymethylcellulose + 0.1-0.5% Tween 80; or

B) 5-20% mannitol + 0.1-0.5% Tween 80; or.

C) 0.5-5% sodium carboxymethylcellulose + 5-20% sorbitol + 0.1-0.5% Tween 80.

The preparation method of sustained-release injection is arbitrary, and several methods can be used for preparation: as, but not limited to, mixing method, melting method, dissolution method, spray drying method to prepare microspheres, dissolution method combined with freeze (dry) pulverization method to make micropowder, Liposome encapsulation method and emulsification method, etc. Among them, the dissolution method (that is, the solvent evaporation method), the drying method, the spray drying method and the emulsification method are preferred. Microspheres can be used to prepare the above-mentioned various sustained-release injections, and the method is arbitrary. The particle size range of the microspheres used may be between 5-400um, preferably between 10-300um, most preferably between 20-200um.

Microspheres can also be used to prepare other sustained-release injections, such as gel injections, gel sustained-release injections, and block copolymer micelle injections. Among them, the block copolymer micelle is formed by hydrophobic-hydrophilic block copolymer in aqueous solution, has a spherical core-shell structure, the hydrophobic block forms the core, and the hydrophilic block forms the shell. The drug-loaded micelles are injected into the body to achieve the purpose of controlled drug release or targeted therapy. The drug carrier used is any one or combination of the above. Wherein preferred molecular weight is polyethylene glycol (PEG) of 1000-15000 as the hydrophilic block of micelle copolymer, preferred biodegradable polymer (such as PLA, polylactide, polycaprolactone and copolymer thereof (molecular weight 1500 -25000)) as the hydrophobic block of the micellar copolymer. The particle size range of the block copolymer micelles can be between 10-300um, preferably between 20-200um. Gel injection is formed by dissolving biodegradable polymers (such as PLA, PLGA or DL-LA and ε-caprolactone copolymer) in certain amphiphilic solvents, and then adding drugs to mix (or suspend) them A fluid gel that can be injected locally. Once injected, the amphiphilic vehicle quickly diffuses into body fluids, and moisture from the body fluids penetrates the gel, solidifying the polymer and slowly releasing the drug.

The present invention finds that the key factor affecting the suspension and/or injection of the drug and/or sustained-release microspheres is the viscosity of the solvent. The greater the viscosity, the better the suspension effect and the stronger the injectability. This unexpected discovery constitutes one of the main index features of the present invention. The viscosity of the vehicle depends on the viscosity of the suspending agent. The viscosity of the suspending agent is 100cp-3000cp (at 20°C-30°C), preferably 1000cp-3000cp (at 20°C-30°C), most preferably 1500cp-3000cp (at 20°C -30°C). The viscosity of the solvent prepared according to this condition is 10cp-650cp (at 20°C-30°C), preferably 20cp-650cp (at 20°C-30°C), most preferably 60cp-650cp (at 20°C-30°C).

There are many methods for the preparation of injections. One is to directly mix the sustained-release microparticles (A) with a suspending agent of "0" in a special solvent to obtain the corresponding sustained-release microparticle injections; the other is to mix the suspending agent without The slow-release microparticles (A) with "0" are mixed in special solvents or ordinary solvents to obtain the corresponding sustained-release microparticle injections; the other is to mix the slow-release microparticles (A) in ordinary solvents, and then add suspending agent Mix evenly to obtain the corresponding sustained-release microparticle injection. In addition, the slow-release microparticles (A) can also be mixed with a special solvent to prepare a corresponding suspension, and then vacuum drying is used to remove the water in the suspension, and then the special solvent or common solvent is used to suspend. The corresponding sustained-release microparticle injection was obtained. The above methods are only for illustration but not limitation of the present invention. It is worth noting that the concentration of suspended drug or sustained-release microspheres (or microcapsules) in the injection depends on specific needs, and can be, but not limited to, 10-400 mg/ml, but preferably 30-300 mg/ml , most preferably at 50-200mg/ml. The viscosity of the injection is 50cp-1000cp (at 20°C-30°C), preferably 100cp-1000cp (at 20°C-30°C), most preferably 200cp-650cp (at 20°C-30°C). This viscosity is suitable for 18-22 gauge needles and special needles with larger inner diameter (up to 3mm).

Sustained-release microspheres can also be used to prepare sustained-release implants, and the pharmaceutical excipients used can be any one or more of the above-mentioned pharmaceutical excipients, but water-soluble polymers are the main choice. Among high molecular polymers, polylactic acid, sebacic acid, mixtures or copolymers of high molecular weight polymers containing polylactic acid or sebacic acid are preferred, and mixtures and copolymers can be selected from, but not limited to, PLA, PLGA , the mixture of PLA and PLGA, the mixture or copolymer of sebacic acid and aromatic polyanhydride or aliphatic polyanhydride. The blending ratio of polylactic acid (PLA) and polyglycolic acid is 10/90-90/10 (weight), preferably 25/75-75/25 (weight). The method of blending is arbitrary. The contents of glycolic acid and lactic acid in copolymerization are respectively 10-90% and 90-10% by weight. The representative of aromatic polyanhydride is p-carboxyphenylpropane (p-CPP), and the content of p-carboxyphenylpropane (p-CPP) and sebacic acid copolymerization is respectively 10-60% and 20-90% by weight, The blending weight ratio is 10-40:50-90, preferably 15-30:65-85.

Therefore, another form of the sustained release agent of the present invention is that the sustained release agent is a sustained release implant. The active ingredient of the anti-tuberculosis implant can be packaged uniformly in the whole pharmaceutical excipient, or in the center of the carrier support or its surface; the active ingredient can be released by direct diffusion and/or polymer degradation.

The feature of the slow-release implant is that the slow-release auxiliary materials used include any one or more of the above-mentioned other auxiliary materials in addition to high molecular polymers. Added pharmaceutical excipients are collectively referred to as additives. Additives can be divided into fillers, porogens, excipients, dispersants, isotonic agents, preservatives, retarders, solubilizers, absorption accelerators, film-forming agents, gelling agents, etc. according to their functions.

The main components of sustained-release implants can be made into various dosage forms. Such as, but not limited to, capsules, sustained-release agents, implants, sustained-release agent implants, etc.; in various shapes, such as, but not limited to, granules, pills, tablets, powders, granules, spherical, Blocky, needle-like, rod-like, columnar and membranous. Among various dosage forms, slow-release implants in vivo are preferred. The volume depends on factors such as the location and size of the lesion. It can be in the form of a rod of 0.1-5 mm (thickness) x 1-10 mm (length), or other shapes such as sheet.

The best dosage form of sustained-release implants is biocompatible, degradable and absorbable sustained-release implants, which can be made into various shapes and dosage forms according to different clinical needs. The packaging method and steps of its main components have been described in detail in U.S. Patent (US5651986), including several methods for preparing sustained-release preparations: such as, but not limited to, (i) mixing the carrier support powder with the drug and then compressing it into Implants, the so-called mixing method; (ii) melting the carrier support, mixing with the drug to be packaged, and then cooling the solid, the so-called melting method; (iii) dissolving the carrier support in a solvent, putting The drug to be packaged is dissolved or dispersed in the polymer solution, and then the solvent is evaporated and dried, which is the so-called dissolution method; (iv) spray drying method; and (v) freeze drying method, etc.

The active ingredients and weight percent of the slow-release implant of the present invention are preferably as follows:

Anti-tuberculosis drugs 2-50%

Sustained release excipients 50-98%

Suspending agent 0.0-30%

The anti-tuberculosis drug and its weight percentage in the anti-tuberculosis sustained-release microspheres of the present invention are preferably as follows:

(1) 5-40% of rifampicin, rifamycin, rifapentine or rifabutin;

(2) Combination of 5-40% rifampicin and 5-40% rifamycin, rifapentine or rifabutin;

(3) Combination of 5-40% rifamycin and 5-40% rifapentine or butin;

(4) Combination of 5-40% rifapentine and 5-40% urbutin.

The weight percent of the sustained-release auxiliary material in the anti-tuberculosis sustained-release microspheres of the present invention is preferably as follows:

(1) 55-90% PLA;

(2) 50-90% PLGA;

(3) 50-85% polyphenylene;

(4) 55-90% difatty acid and sebacic acid copolymer; or

(5) 55-90% EVAc.

In addition, the selected auxiliary materials can also be a combination of any one or more of the above.

The invention can be used to prepare pharmaceutical preparations for treating various tuberculosis of humans and animals, mainly slow-release injections or slow-release implants. Although the prepared pharmaceutical preparation can be used for the treatment of systemic tuberculosis such as pulmonary tuberculosis, it is preferred to treat local lesions. Common local lesions or extrapulmonary tuberculosis mainly include: tuberculosis, lymphatic tuberculosis, bone and joint tuberculosis, synovial tuberculosis, tuberculous osteomyelitis, renal tuberculosis, skin tuberculosis, intestinal tuberculosis, breast tuberculosis, genital tuberculosis (fallopian tube, Endometrium, testis, epididymis), anal tuberculosis, thyroid tuberculosis, pericardial tuberculosis, chest wall tuberculosis, tuberculosis fistula, pleural tuberculosis, etc. In addition, local lesions also include chronic fibrous cavitary tuberculosis and chronic lesions caused by or combined with tuberculosis, such as: but not limited to, severe decubitus ulcers, intractable skin ulcers, diabetic foot, femoral head necrosis, and senile prostate diseases wait.

The route of administration depends on many factors. In order to obtain an effective concentration at the site of tuberculosis, the drug can be administered through various routes, such as subcutaneous, intracavitary (such as intraperitoneal, thoracic and spinal canal), perilesional or intralesional injection or placement , intralymph node and intramedullary injection, but the slow-release implant is preferably injected locally (slow-release injection) or placed (slow-release implant) in the lesion. It can be injected or placed during the operation or before and after the operation; it can be used for interventional therapy through fiberoptic bronchoscope and other instruments, such as the treatment of pulmonary tuberculosis cavity; it can also be used for percutaneous puncture into the lesion for interventional therapy; it can also be injected or placed in the joint cavity; it can be used with the whole body Chemotherapy can be applied at the same time or separately before and after, but it is better to have systemic anti-tuberculosis treatment for one week to several months before and after local application.

The dosage varies due to the composition of the drug, but the total amount of the drug can range from 10% to 200% of the daily dose of the conventional route. When two drugs are used in combination, the dose of each drug should not exceed 100% of its daily dose by conventional route. If the lesion has not been completely cleared or improved, re-placement or injection of sustained-release agents may be considered after 20 to 40 days. In order to prevent the spread of tuberculosis in the lesion, systemic administration should be combined before and after each local administration.

The sustained-release injection or sustained-release implant prepared by the present invention can also add other medicinal ingredients, such as, but not limited to, antibiotics, painkillers, anticoagulants, hemostatic drugs and the like.

Technical method of the present invention is described further by following test and embodiment:

Experiment 1. Comparison of local drug concentrations after different application of anti-tuberculosis drugs (rifampicin)

Rats were taken as the test object, and they were divided into groups to receive the same amount of rifampicin (10 mg) in the following different ways: group 1, intraperitoneal injection of common rifampicin injection; group 2, quarter rib subcutaneous injection of common rifampicin Injection; group 3, subcutaneous injection of rifampicin sustained-release injection in quarter rib; group 4, subcutaneous rifampicin sustained-release implant in quarter rib. After one week, two weeks, and three weeks, the drug concentration at the local administration site was measured respectively. The results show that there are significant differences in the local drug concentration after application in different ways, and local administration can be significantly improved, and the effective drug concentration at the administration site can be effectively maintained. Among them, local placement of slow-release implants and injection of slow-release injections have the best results. However, local injection of sustained-release injections is the most convenient and easy to operate. This finding constitutes an important feature of the present invention. The following related experiments further confirmed this point.

Experiment 2. Comparison of antibacterial effects in vivo after different application of anti-tuberculosis drugs

Rats were taken as the test subjects, 2×10 ⁵ Staphylococcus aureus were injected into the bone marrow cavity of their femurs, and after one week they were given equal doses of rifampicin according to the grouping in Experiment 1 (10 rats/group). Subsequently, local redness and swelling and other inflammatory changes were checked. Thirty days later, the animals were sacrificed and the local bone marrow was collected for bacterial examination. The results showed that the injection of rifampicin sustained-release injection and rifampicin sustained-release implant group had the best effect, and the local redness and swelling began to shrink significantly in the first week after treatment, and no animals died. In the intraperitoneal injection of common rifampicin injection group, 80% of the animals died within 20 days; in the local injection of common rifampicin injection group, 20% of the animals died within 20 days, but 60% of the animals died within 30 days. Comparing the antibacterial effects shows that the effects of different methods of application are significantly different, and local administration can significantly increase and effectively maintain the effective drug concentration at the site, among which local slow-release implants and injection slow-release injections have the best effect . However, the injection of sustained-release injections is the most convenient and easy to operate. Not only the curative effect is good, but also the side effects are small.

Experiment 3. Comparison of antibacterial effects of rifampicin in different ways in vivo

Taking rats as test objects, repeat test 2, the results are shown in Table 1

Table 1

Test group (n) received treatment Animal Mortality (%) P value 1 (10) control 0 2 (10) Ordinary rifampicin injection intraperitoneally 100 3 (10) Local injection of common rifampicin injection 70 <0.05 4 (10) Local injection of rifampicin sustained release injection 10 <0.01 5 (10) Locally placed rifampicin extended-release implant 0 <0.01

The above results show that the antibacterial effect of the anti-tuberculosis drug rifampicin is different when administered through different routes, and the effect of local application is better (P<0.05). The effect of releasing the implant is better.

Experiment 4. Comparison of antibacterial effects of rifampicin in different ways

Taking white rats as the test object, repeat test 3, but observe the animals until 60 days after administration, the results are shown in Table 2

Table 2

Test group (n) received treatment Animal Mortality (%) P value 1 (10) control 0 2 (10) Ordinary rifampicin injection intraperitoneally 100 3 (10) Local injection of common rifampicin injection 100 4 (10) Local injection of rifampicin sustained release injection 60 5 (10) Locally placed rifampicin extended-release implant 20 <0.01

The above results show that the anti-bacterial effect of rifampicin, an anti-tuberculosis drug administered through different routes, is different, and the effect of local application is better. Better results. But the effect of local rifampicin sustained-release implant was better than local injection of rifampicin sustained-release injection (P<0.01).

The same obvious therapeutic effect is also seen in locally placed and locally injected rifampicin analog sustained-release agents: rifamycin, rifapentine and utilbutin.

Experiment 5. The synergistic effect of different drug combinations

Get 2 * ¹⁰ Mycobacterium tuberculosis culture 24 hours and treat with same concentration (5ug/ml) different drugs for 24 hours, detect cell growth inhibition rate (%), the results are shown in Table 3

Table 3 Synergistic effect of different drug combinations (inhibition rate%)

The results of experiment 5 showed that the anti-tuberculosis drug rifampicin and rifampicin analogs had obvious antibacterial effect when used alone, but the effect of combined application was better (P<0.01).

In conclusion, both local placement and local injection of rifampicin or rifampicin analog slow-release preparations have obvious inhibitory effects on bacterial growth, and the significant therapeutic effect shown is related to the effective drug concentration obtained locally. Therefore, the active ingredient of the sustained-release preparation of the present invention is rifampicin or any rifampicin analogue. The rifampicin analog used may be, but not limited to, rifamycin, rifapentine and utilbutin.

Drugs containing the above active ingredients can be made into sustained-release microspheres, and then made into sustained-release injections and implants, among which suspension injections formed by combining with special solvents containing suspending agents are preferred.

The sustained-release injection or sustained-release implant can also be further illustrated by the following embodiments. The above-mentioned embodiment and the following embodiment are only to further illustrate the present invention, but not to limit its content and use in any way.

(4) Specific implementation methods

Example 1.

Put 90, 90 and 80 mg of polyphenylpropane (p-carboxyphenylpropane (p-CPP): sebacic acid (SA) at 20:80) copolymers into (A), (B) and (C) three Add 100 ml of dichloromethane to each container, dissolve and mix well, add 10 mg rifampicin, 10 mg rifamycin, 10 mg rifampicin and 10 mg rifamycin respectively, re-shake and spray dry Microspheres for injection containing 10% rifampicin, 10% rifamycin and 10% rifampicin and 10% rifamycin were prepared by method. Then suspend the microspheres in physiological saline containing 15% mannitol to prepare the corresponding suspension-type sustained-release injection. The viscosity of the injection is 300cp-600cp (at 20°C-30°C). The drug release time of the sustained release injection in physiological saline in vitro is 15-20 days, and the drug release time in mice subcutaneous is about 30-40 days.

Example 2.

The method step of being processed into slow-release injection is identical with embodiment 1, but difference is that contained anti-tuberculosis active ingredient and weight percentage thereof are:

(a) 2-40% rifampicin, rifamycin, rifapentine or utilbutin;

(b) 2-40% rifampicin in combination with 2-40% rifamycin, rifapentine or rifabutin;

(c) 2-40% rifamycin in combination with 2-40% rifapentine or rifabutin;

(d) Combination of 2-40% rifapentine and 2-40% urbutin.

Example 3.

Put 70 mg of polylactic acid (PLGA, 75:25) with a peak molecular weight of 65,000 into three containers (A), (B) and (C) respectively, and then add 100 ml of dichloromethane to each, dissolve and mix well , add 30mg rifampicin, 30mg rifapentine, 15mg rifampicin and 15mg rifapentine into three containers respectively, shake up again and use spray drying method to prepare 30% rifampicin, 30% rifapentine 15% rifampicin and 15% rifapentine injection microspheres. The dried microspheres are suspended in physiological saline containing 1.5% sodium carboxymethylcellulose to prepare the corresponding suspension-type sustained-release injection. The viscosity of the injection is 300cp-600cp (at 20°C-30°C). The drug release time of the slow-release injection in physiological saline in vitro is 10-15 days, and the drug release time in mice subcutaneous is about 20-30 days.

Example 4

The method step of being processed into slow-release injection is identical with embodiment 3, but difference is that contained anti-tuberculosis active ingredient and weight percentage thereof are:

(a) 2-40% rifampicin, rifamycin, rifapentine or utilbutin;

(b) 2-40% rifampicin in combination with 2-40% rifamycin, rifapentine or rifabutin;

(c) 2-40% rifamycin in combination with 2-40% rifapentine or rifabutin;

(d) Combination of 2-40% rifapentine and 2-40% urbutin.

Example 5.

Put 70mg of ethylene vinyl acetate copolymer (EVAc) into a container, add 100ml of dichloromethane to dissolve and mix well, add 20mg of rifampicin and 10mg of utilbutin, re-shake and use spray drying method to prepare 20 % rifampicin and 10% microspheres for injection utilizing butin. Then suspend the microspheres in the injection solution containing 5-15% sorbitol to prepare the corresponding suspension sustained-release injection. The drug release time of the slow-release injection in physiological saline in vitro is 10-15 days, and the drug release time in mice subcutaneous is about 20-30 days.

Example 6.

The method step of being processed into slow-release injection is identical with embodiment 5, but difference is that contained anti-tuberculosis active ingredient is:

Combination of 10-20% Utilbutin with 10-20% rifampicin, rifamycin or rifapentine.

Example 7.

Put 70mg of polyphenylpropane (p-CPP: 20:80 of sebacic acid (SA)) copolymer into a container, add 100ml of dichloromethane, dissolve and mix well, then add 20mg Rifamycin and 10 mg rifapentine were re-shaken and spray-dried to prepare microspheres for injection containing 20% rifamycin and 10% rifapentine. Then suspend the microspheres in physiological saline containing 1.5% sodium carboxymethylcellulose and 0.5% Tween 80 to prepare the corresponding suspension-type sustained-release injection. The drug release time of the slow-release injection in physiological saline in vitro is 10-15 days, and the drug release time in mice subcutaneous is about 20-30 days.

Example 8.

The method step of being processed into sustained-release injection is the same as in Example 7, but the difference is that the contained anti-tuberculosis active ingredients are:

Combination of 10-20% rifapentine with 10-20% rifampicin, rifamycin, or ubutin.

Example 9

Put 70mg of polyphenylpropane (p-CPP: 20:80 of sebacic acid (SA)) copolymer into a container, add 100ml of dichloromethane, dissolve and mix well, then add 15mg Rifapentine and 15 mg rifampicin were re-shaken and spray-dried to prepare microspheres for injection containing 15% rifapentine and 15% rifampicin. Then the microspheres were suspended in physiological saline containing 1.5% sodium carboxymethylcellulose, 15% sorbitol and 0.2% Tween 80 to prepare the corresponding suspension-type sustained-release injection. The drug release time of the slow-release injection in physiological saline in vitro is 10-15 days, and the drug release time in mice subcutaneous is about 20-30 days.

Example 10

The method step of being processed into sustained-release injection is the same as in Example 9, but the difference is that the contained anti-tuberculosis active ingredients are:

Combination of 15% rifampicin with 15% rifapentine, rifamycin or utilibutin.

Example 11

Put 70mg of polyphenylpropane (p-CPP: 20:80 of sebacic acid (SA)) copolymer into a container, add 100ml of dichloromethane, dissolve and mix well, then add 10mg Rifampicin and 20mg rifapentine were re-shaken and spray-dried to prepare microspheres for injection containing 10% rifampicin and 20% rifapentine. Then the microspheres are pressed into tablets to prepare the corresponding slow-release implants. The drug release time of the slow-release implant in physiological saline in vitro is 10-15 days, and the drug release time in mice subcutaneous is about 30-40 days.

Example 12

The method step of being processed into slow-release implants is the same as in Example 11, but the difference is that the contained anti-tuberculosis active ingredients are:

Combination of 10-20% rifampicin with 10-20% rifapentine, rifamycin or ubutin.

Example 13

Put 70 mg of polylactic acid (PLGA, 50:50) with a peak molecular weight of 45,000 into a container, add 100 ml of dichloromethane, dissolve and mix well, add 10 mg of rifampicin and 20 mg of rifapentine, shake well and spray Microspheres for injection containing 10% rifampicin and 20% rifapentine were prepared by drying method. Then the microspheres are pressed into tablets to prepare the corresponding slow-release implants. The drug release time of the sustained release implant in physiological saline in vitro is 15-25 days, and the drug release time in mice subcutaneous is about 35-50 days.

Example 14

The method step of being processed into slow-release implant is identical with embodiment 11,13, but difference is that contained anti-tuberculosis active ingredient and weight percent are:

(a) 2-40% rifampicin, rifamycin, rifapentine or utilbutin;

(b) 2-40% rifampicin in combination with 2-40% rifamycin, rifapentine or rifabutin;

(c) 2-40% rifamycin in combination with 2-40% rifapentine or rifabutin;

(d) Combination of 2-40% rifapentine and 2-40% urbutin.

Example 15

The method step of being processed into slow-release preparation is identical with embodiment 1-14, but difference is that the slow-release auxiliary material used is one of following or its combination:

a) Polylactic acid (PLA) with a peak molecular weight of 10,000-30,000, 30,000-60,000, 60,000, 100,000 or 100,000-150,000;

b) A copolymer of polyglycolic acid and glycolic acid (PLGA) with a peak molecular weight of 10,000-30,000, 30,000-60,000, 60,000-100,000 or 100,000-150,000, wherein the ratio of polyglycolic acid to glycolic acid is 50-95:50 -50;

c) ethylene vinyl acetate copolymer (EVAc);

d) 10:90, 20:80, 30:70, 40:60, 50:50, or 60:40 p-carboxyphenylpropane (p-CPP):sebacic acid (SA) copolymer (polyphenylpropane );

e) difatty acid and sebacic acid copolymer;

f) poly(erucic acid dimer-sebacic acid) copolymer;

g) poly(fumaric acid-sebacic acid) copolymer;

h) Xylitol, oligosaccharides, chondroitin, chitin, potassium salts, sodium salts, hyaluronic acid, collagen, gelatin or albumin.

Example 16

The method step of being processed into sustained-release injection is identical with embodiment 1-10, but difference is that used suspending agent is respectively following one or its combination:

a) 0.5-3.0% carboxymethylcellulose (sodium);

b) 5-15% mannitol;

c) 5-15% sorbitol;

d) 0.1-1.5% surface active substances;

e) 0.1-0.5% Tween 20;

f) (iodo)glycerin, simethicone, propylene glycol or carbomer;

g) 0.5-5% sodium carboxymethylcellulose + 0.1-0.5% Tween 80;

h) 5-20% mannitol + 0.1-0.5% Tween 80; or

i) 0.5-5% sodium carboxymethylcellulose + 5-20% sorbitol + 0.1-0.5% Tween 80.

The above embodiments are for illustration only, and do not limit the application of the present invention.

See the claims for the disclosed and protected contents of the present invention.

Claims (3)

1. slow released antituberculotic preparation is characterized in that this slow released antituberculotic preparation is a slow releasing injection, is grouped into by following one-tenth:

(1) the tuberculosis effective ingredient is that 10% rifampicin and 10% rifamycin, slow-release auxiliary material are to carboxy phenyl propane in the sustained-release micro-spheres: decanedioic acid is 20: 80 a polifeprosan, solvent is the normal saline that contains 15% mannitol, in the time of 20 ℃-30 ℃, the viscosity of injection is 300cp-600cp; Or

(2) the tuberculosis effective ingredient is that 15% rifampicin and 15% rifapentine, slow-release auxiliary material are 65000 polylactic acid for the molecular weight peak value in the sustained-release micro-spheres: hydroxyacetic acid is 75: 25 polylactic acid and a co-glycolic acid, solvent is the normal saline that contains 1.5% sodium carboxymethyl cellulose, in the time of 20 ℃ 30 ℃, the viscosity of injection is 300cp-600cp;

Sustained-release micro-spheres is weight percentage.

2. the tuberculosis slow releasing injection according to claim 1 is characterized in that the sustained-release micro-spheres in the tuberculosis slow releasing injection also is used to prepare sustained-release implant.

3. slow released antituberculotic preparation is characterized in that this slow released antituberculotic preparation is the tuberculosis sustained-release implant, is grouped into by following one-tenth:

(1) the tuberculosis effective ingredient is 10% rifampicin and 20% rifapentine, and slow-release auxiliary material is to carboxy phenyl propane: decanedioic acid is 20: 80 a polifeprosan; Or

(2) the tuberculosis effective ingredient is 10% rifampicin and 20% rifapentine, and slow-release auxiliary material is 45000 polylactic acid for the molecular weight peak value: hydroxyacetic acid is 50: 50 polylactic acid and a co-glycolic acid;

Below all be weight percentage.