WO2017114095A1 - 可利霉素在抗结核分枝杆菌感染中的应用 - Google Patents

可利霉素在抗结核分枝杆菌感染中的应用 Download PDF

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WO2017114095A1
WO2017114095A1 PCT/CN2016/108502 CN2016108502W WO2017114095A1 WO 2017114095 A1 WO2017114095 A1 WO 2017114095A1 CN 2016108502 W CN2016108502 W CN 2016108502W WO 2017114095 A1 WO2017114095 A1 WO 2017114095A1
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tuberculosis
mycobacterium tuberculosis
colimycin
drugs
kelimycin
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PCT/CN2016/108502
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English (en)
French (fr)
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王以光
姜洋
赵小峰
郝卫清
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沈阳同联集团有限公司
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Priority to MYPI2018001121A priority Critical patent/MY191800A/en
Priority to ES16880885T priority patent/ES2785644T3/es
Application filed by 沈阳同联集团有限公司 filed Critical 沈阳同联集团有限公司
Priority to BR112018013327-2A priority patent/BR112018013327A2/zh
Priority to RU2018125622A priority patent/RU2733382C2/ru
Priority to PL16880885T priority patent/PL3384915T3/pl
Priority to EP16880885.5A priority patent/EP3384915B1/en
Priority to KR1020187021578A priority patent/KR20180098624A/ko
Priority to JP2018553283A priority patent/JP6770584B2/ja
Priority to UAA201808048A priority patent/UA121159C2/uk
Priority to PE2018001212A priority patent/PE20190210A1/es
Priority to US16/067,327 priority patent/US11000708B2/en
Priority to DK16880885.5T priority patent/DK3384915T3/da
Priority to CA3010020A priority patent/CA3010020A1/en
Priority to AU2016382584A priority patent/AU2016382584B2/en
Publication of WO2017114095A1 publication Critical patent/WO2017114095A1/zh
Priority to PH12018501394A priority patent/PH12018501394A1/en
Priority to ZA2018/04398A priority patent/ZA201804398B/en
Priority to CONC2018/0007133A priority patent/CO2018007133A2/es

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • A61P31/06—Antibacterial agents for tuberculosis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0053—Mouth and digestive tract, i.e. intraoral and peroral administration
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Definitions

  • the invention relates to the use of macrolide antibiotics in the treatment of Mycobacterium tuberculosis infection.
  • Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis (MTB) infection, which mainly affects the lungs. It is the single most common cause of death among infectious diseases. Tuberculosis is more common in immunocompromised populations and is the most common AIDS-related opportunistic infection. According to the World Health Organization (WHO), there are 8 million to 10 million new cases of tuberculosis every year in the world, and 3 to 4 million people die of tuberculosis, and they are more likely to be in developing countries. It is also predicted that about 1 billion people will be infected and 3-5 million will die of tuberculosis between 2000 and 2020.
  • WHO World Health Organization
  • MTB resistance has increased year by year and will become a major threat to global tuberculosis control.
  • China is one of the 22 countries with high incidence of tuberculosis in the world.
  • the number of active tuberculosis patients ranks second in the world, and the epidemic characteristics are high infection rate, high prevalence rate, high drug resistance rate and high mortality rate. 2 million MTB nationwide. More than a quarter of the positive patients were tuberculosis-resistant patients.
  • the first-line drugs commonly used in the treatment of tuberculosis include: rifampicin, isoniazid, streptomycin, ethambutol and pyrazinamide.
  • these drugs have many adverse reactions, limited bactericidal effects, and long course of treatment, usually used for more than 6 months, and poor patient compliance.
  • Second-line anti-tuberculosis drugs include: capreomycin, ethionamide, p-aminosalicylic acid, cycloserine, ciprofloxacin, amikacin and kanamycin Etc., and these drugs have large adverse reactions, longer treatment time (18 to 24 months), expensive, and lower cure rate. It is noteworthy that clinical studies have shown that first-line anti-tuberculosis drugs can cause damage to the liver.
  • the adverse reactions of isoniazid are peripheral neuritis, hepatotoxicity, central nervous system disorders and allergic reactions; the adverse reactions of rifampicin are Hepatotoxicity, digestive tract discomfort, nervous system symptoms and allergic reactions; adverse reactions of ethambutol are mainly optic neurotoxicity; pyrazinamide adverse reactions are yellowing of skin and increased blood uric acid; streptomycin toxicity and bacterial resistance Sexual problems severely limit their use, because the combination with other drugs can reduce the incidence of bacterial resistance, although not much clinical application, but still used as a first-line drug against tuberculosis. [Zhu Shanmei Strait Pharmacy 2010, 22(2): 123-125].
  • the new macrolides clarithromycin, azithromycin and roxithromycin are derivatives of fourteen-membered erythromycin, and their antibacterial mechanism is different from that of the first-line and second-line anti-tuberculosis drugs.
  • the 50S subunit reversibly binds to interfere with protein synthesis. It has been reported in China that the MIC of clarithromycin against sensitive M. tuberculosis is 0.25-2.0 ⁇ g/ml, the MIC of resistant bacteria is 2.0-32 ⁇ g/ml, and the MIC of azithromycin against M.
  • tuberculosis is 128 ⁇ g/ ML [Tang Shenjie, the latest advances in anti-tuberculosis drug research, China National Defense Magazine, 2006 28-volume supplement 1-3 pages].
  • the MIC of clarithromycin against M. tuberculosis H37Rv (ATCC 27294) is 6 ⁇ g/ml; the MIC of azithromycin is 95 ⁇ g/ml [Kanakeshwari Falzari et al: In vitro and in vivo activities o macrolide derivatives against Mycobacterium tuberculosis Antimicrob. Agents and Chemother. 2005, 49(4): 1447-1454]; MIC of roxithromycin ⁇ 64 ⁇ g/ml.
  • the test results show that colimycin is effective against Gram-positive bacteria, especially some resistant bacteria (such as ⁇ -lactam-resistant Staphylococcus aureus, erythromycin-resistant Staphylococcus aureus, etc.), and has no obvious cross with similar drugs.
  • the main steps of the present invention are to determine the activity of colimycin against Mycobacterium tuberculosis by using the absolute concentration method for the clinically isolated Mycobacterium tuberculosis, and to use the anti-tuberculosis first-line drugs isoniazid and rifampicin as clinical controls.
  • the results showed that there were 172 strains of colimycin in the clinical isolates of 240 strains of Mycobacterium tuberculosis, the total effective rate was 71.66%, and 37 of them were better than isoniazid, accounting for the effective number of bacteria. 21.5%; 39 strains with better activity than rifampicin, accounting for 22.7% of the effective bacteria; 23 strains with better activity than both, accounting for 13.4% of the effective bacteria count.
  • the results of the present invention suggest that colimycin is expected to exploit new uses in the treatment of M. tuberculosis infections resistant to isoniazid and rifampicin.
  • the invention also provides the use of a composition comprising colimycin as an active ingredient and a pharmaceutically acceptable carrier for the preparation of a medicament for the prevention of tuberculosis infection.
  • the use of the present invention may be in the form of an oral dosage form, or an injectable dosage form or any other suitable dosage form.
  • the first-line drug related to tuberculosis commonly used in clinical practice is used as a control.
  • a series of studies on the activity of lixinomycin against Mycobacterium tuberculosis showed that the effective number of the colimycin showed activity in clinically isolated Mycobacterium tuberculosis was superior to the control group. It is used in the treatment of certain drug-resistant Mycobacterium tuberculosis infections.
  • PBS pH 6.8 was added to 40 mL, and centrifuged at 3000 g for 20 minutes, and then the supernatant was discarded to leave a precipitate. 2 mL of PBS (pH 6.8) was added to prepare a suspension. The treated specimen inoculated medium was subjected to solid culture.
  • composition of the medium is shown in Table 1.
  • the ingredients were added to distilled water at the doses listed and fully dissolved; boiled for 30 minutes or at 121 ° C for 15 minutes.
  • Mycobacterium tuberculosis culture-positive bacteria for a little moss, apply it evenly on the glass slide, fix it on the flame, dye it with stone carbonate reddish solution, heat on the flame for 5 minutes, decolorize with 3% hydrochloric acid alcohol for about 1 minute, rinse with water, then After counterstaining with alkaline melanin solution for 1 minute, rinse with water, absorb water, and observe under oil microscope.
  • the stained red is Mycobacterium tuberculosis.
  • Colimycin standard product from China National Institute for the Control of Pharmaceutical and Biological Products; reference drug: isoniazid, rifampicin using Sigma standard products.
  • Anti-tuberculosis drugs are formulated into mother liquor at a certain concentration, and then added to the medium in a certain amount to prepare the required dose (Table 2).
  • the strains isolated from the clinical specimens were smeared as cultures of acid-fast bacteria, diluted with physiological saline containing 10% Tween 80, and turbid with McFarland standard turbidity tube (Guangdong Huan Kai Microbiology Co., Ltd.), preparation 10 - 2 mg/mL of bacterial solution was inoculated into the medium containing the test drug.
  • Negative and positive controls were set for each batch of trials.
  • the negative control was unmedicated medium.
  • the positive control was M. tuberculosis standard strain H37Rv.
  • the inoculum size per tube was 0.1 mL.
  • the cultured medium after inoculation was cultured at 37 °C.
  • the bacterial liquid was observed once on the third day after inoculation, and then observed once a week, and the results were reported in 4 weeks.
  • Control no drug medium. All isolated strains grew 4+ in drug-free medium.
  • Colimycin is superior to 250 ⁇ g/ml of rifampicin in certain clinical Mycobacterium tuberculosis activities.
  • the activity of colimycin was compared with the clinical activity of Mycobacterium tuberculosis resistant to isoniazid and rifampicin. The results showed that the activity of colimycin 20 ⁇ g/ml was better than that of 23 strains of Mycobacterium tuberculosis. ⁇ and 250 ⁇ g / ml rifampicin (Table 6).
  • kalliktomycin is not only active against the clinical first-line anti-tuberculosis drugs isoniazid and rifampicin-sensitive bacteria, but also active against some isoniazid and rifampicin-resistant bacteria, and is expected to be clinically New applications have been made in the treatment of drug-resistant M. tuberculosis infections.

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Abstract

可利霉素在制备抗结核杆菌感染中的应用，其主要步骤是，以临床一线抗结核药异烟肼和利福平作为对照，采用绝对浓度法进行可利霉素抗结核分枝杆菌活性测定，结果表明，可利霉素对临床分离的结核分枝杆菌包括耐药菌的活性，明显优于临床一线对照药异烟肼和利福平，有望发掘出可利霉素在制备治疗结核菌感染疾病药物中的用途。

Description

可利霉素在抗结核分枝杆菌感染中的应用 技术领域：
本发明涉及大环内酯类抗生素在治疗结核杆菌感染中的应用。
技术背景：
结核病是由结核分枝杆菌(MTB)感染引起的慢性传染病，主要累及肺，是传染病中单一导致死亡人数最多的疾病。结核病多见于免疫功能低下人群，也是最常见的与艾滋病相关的一种机会感染性疾病。据世界卫生组织(WHO)报告，全世界每年有800万-1000万肺结核新发病例，有300万～400万人死于结核病，且以发展中国家患者为甚。另据预测，2000-2020年将有约10亿人被感染，3500万人死于结核病。与此同时，MTB耐药性逐年增加，将成为全球控制结核病的一大威胁。我国是全球结核病高发的22个国家之一，活动性结核病患者数居世界第二位，且流行的特点呈现高感染率、高患病率、高耐药率和高死亡率，全国200万MTB阳性患者中，有1/4以上为结核菌耐药患者。
目前，临床常用治疗结核病的一线药物(具有明确的抗结核分枝杆菌活性)有：利福平、异烟肼、链霉素、乙胺丁醇和吡嗪酰胺等5种。然而这些药物不良反应多，杀菌作用有限，疗程较长，通常使用6个月以上，患者依从性差。二线抗结核药物(对结核分枝杆菌具抑菌作用)有：卷曲霉素、乙硫异烟胺、对氨基水杨酸、环丝氨酸、环丙沙星、阿米卡星和卡那霉素等，而这些药物不良反应较大，治疗时间更长(18～24个月)，花费昂贵，治愈率也更低。值得关注的是，临床研究表明，一线抗结核药物均可对肝脏造成损害，如异烟肼的不良反应为周围神经炎、肝毒性、中枢神经系统障碍及过敏反应；利福平的不良反应有肝毒性、消化道不适、神经系统症状和过敏反应；乙胺丁醇的不良反应主要为视神经毒性；吡嗪酰胺的不良反应为出现皮肤黄染和血尿酸增高；链霉素毒性及细菌耐药性问题严重限制了其应用，因与其它药物联用可减少细菌耐药性的发生率，尽管临床应用不多，但仍用作抗结核病的一线药物。[朱珊梅海峡药学2010，22(2)：123-125]。另据报道，2010年，对518株临床分离结核分枝杆菌的研究表明，在 一线药物中，对异烟肼的耐药率最高达到53.67％，对链霉素耐药率为45.95％；在二线药物中，对氧氟沙星耐药率达39.77％，对阿米卡星耐药率为15.83％，对卷曲霉素耐药率为21.81％；在321株对一种及以上一线药物的耐药菌株中，对氧氟沙星耐药高达57.01％，对阿米卡星和卷曲霉素耐药率分别为25.55％和33.02％；在217株多重耐药株中，对氧氟沙星耐药率达72.35％[刘一典等2010中国防痨协会临床基础专业学术大会汇编274-275页]。这对我国结核病控制工作，特别是耐多药结核病控制工作的实施带来严峻的挑战。因此，尽快寻找抗结核分支杆菌特别是抗多重耐药菌株的替代产品已经刻不容缓了。
新大环内酯类药物克拉霉素、阿奇霉素、罗红霉素为十四元环红霉素的衍生物，其抗菌作用机制与上述一线、二线抗结核药物不同，是与菌体内核糖体的50S亚基可逆性结合，干扰蛋白质的合成。国内有报道称，克拉霉素对敏感结核分枝杆菌的MIC为0.25-2.0微克/毫升，对耐药菌的MIC为2.0-32微克/毫升；阿奇霉素抗结核分枝杆菌的MIC为128微克/毫升[唐神结，抗结核药物研究最新进展，中国防痨杂志，2006年28卷增刊1-3页]。据国外报道，克拉霉素对M.tuberculosis H37Rv(ATCC 27294)的MIC为6微克/毫升；阿奇霉素的MIC为95微克/毫升[Kanakeshwari Falzari et al:In vitro and in vivo activities o macrolide derivatives against Mycobacterium tuberculosis.Antimicrob.Agents and Chemother.2005,49(4):1447-1454]；罗红霉素的MIC≥64微克/毫升。此类药物并未被列入临床治疗结核分枝杆菌感染的药物，但有临床上单独或与抗结核药物合并使用治疗耐多药结核菌感染疾病的报道[许丽等深圳市住院病人抗结核相关抗菌药物应用调查中国防痨杂志2010，32(3)：151-154]。
研究证明，16元环与14元环大环内酯类抗生素具有相类似的抗菌作用机制。此类药物对结核分枝杆菌活性低，如对M.tuberculosis H37Rv(ATCC 27294)，泰洛菌素的MIC为58.6微克/毫升，螺旋霉素和麦迪霉素均>100微克/毫升[Kanakeshwari Falzari et al:In vitro and in vivo activities of macrolide derivatives against Mycobacterium tuberculosis.Antimicrob.Agents and Chemother.2005,49(4):1447-1454]。因此，迄今为止，国内外尚无此类抗生素用于临床治疗结核病的相关报道。
本实验室研制的一种新型16元环大环内酯类抗生素-可利霉素(原用名生技 霉素、必特螺旋霉素)，系以异戊酰螺旋霉素为主组分的4”-酰化螺旋霉素，其作用机制是通过与细菌核糖体结合而抑制其蛋白质合成。体内外试验结果表明，可利霉素对革兰阳性菌，尤其对某些耐药菌(如耐β-内酰胺金葡菌、耐红霉素金葡菌等)有效，与同类药无明显的交叉耐药性。与此同时，它对支原体、衣原体有很好的抗菌活性，对部分革兰阴性菌也有抗菌作用，且对弓形体、军团菌等亦有良好抗菌效果[王以光等，“必特螺旋霉素及其在抗感染性疾病中的应用”，2003年12月23日，中国发明专利：ZL 2003 1 0122420.9]。可利霉素具有良好的组织渗透性，其体内抗菌活性明显优于体外，且具有潜在的免疫调节作用。通过I、II、III期临床研究，证明可利霉素是一种使用安全疗效显著的抗生素。本实验室进一步发掘可利霉素潜在功效，扩大可利霉素临床适应症和使用范围。
发明内容：
本发明的目的是，提供可利霉素对临床分离结核分枝杆菌活性的系列检测实验研究，以证明其可能具有治疗结核杆菌感染疾病的新用途。
本发明的主要步骤是，采用绝对浓度法对临床分离的结核杆菌，进行可利霉素抗结核分枝杆菌活性测定，并以临床使用的抗结核病一线药物异烟肼和利福平作为对照。实验结果表明，可利霉素对临床分离的240株结核分枝杆菌中显示活性的菌株为172株，总有效率为71.66％，其中活性优于异烟肼的有37株，占有效菌数的21.5％；活性优于利福平的有39株，占有效菌数的22.7％；活性比两者均优的有23株，占有效菌数的13.4％。本发明结果提示，可利霉素有望在治疗对异烟肼和利福平耐药的结核分枝杆菌感染疾病中发掘新的用途。
本发明还提供可利霉素为有效成分与药学上可接受的载体组成的组合物在制备抗结核菌感染药物中的应用。
本发明所述的应用，可以采用口服剂型，也可以采用注射剂型或其它任何适宜的剂型。
实施方案：
以下实施例仅为帮助本领域技术人员更好地理解本发明，但不以任何方式限制本发明。
在具体实施方式中，以临床常用的抗结核病相关一线药物为对照，进行了可 利霉素抗结核分枝杆菌活性测定的系列研究，结果表明，所述可利霉素对临床分离的结核分枝杆菌中显示活性的菌株有效数，均优于对照组。其在治疗某些耐药结核分支杆菌感染疾病中得到应用。
《实施例1》结核分枝杆菌标本的获取与处理
根据卫生部发布的国标WS288-2008《肺结核诊断标准》的规定，挑选临床表现症状、体征及胸部影像学检查确诊或高度疑似的结核患者。收集被选患者痰、胸水、脑脊液、脓汁标本约2mL，加入带有螺旋帽的50mL离心管中。加入与标本等量的N-乙酰-L-半胱氨酸氢氧化钠(NaOH-NALC)前处理液，漩涡振荡20秒。室温静置18分钟。加入PBS(pH6.8)至40mL，3000g离心20分钟，然后弃上清，留沉淀。加入2mL PBS(pH6.8)，以制备成悬浊液。处理后的标本接种培养基进行固体培养。
《实施例2》结核分枝杆菌标本的分离培养鉴定
1.培养基的配制：
培养基成分如表1所示。按其所列剂量将各成分加入蒸馏水，充分溶解；煮沸30分钟或121℃高压15分钟。
取新鲜鸡卵，经自来水清洗，肥皂水刷洗干净，待干后以75％酒精擦拭消毒。在无菌操作下将卵液倒入已灭菌的有刻度搪瓷杯内，充分搅拌混匀，灭菌纱布过滤后，取1000mL加入，充分混匀；加入2％孔雀绿20mL，充分混匀；分装试管(18mm×180mm)内加入培养基7mL，置蒸汽恒温箱内85℃50分钟凝固。按5％取管制备好的培养基，置入37℃培养48小时，进行无菌检验；无菌检验合格后，放置4℃冰箱备用，一个月内使用。
表1培养基
Figure PCTCN2016108502-appb-000001
2.结核杆菌的接种培养：
吸取《实施例1》处理好的标本0.1mL。将其均匀接种于培养基斜面上，混悬液应布满整个斜面。放37℃孵箱内培养。第3天，第7天观察细菌生长情况，其后每周观察一次，但若发现有阳性可疑者，应及时挑出，每天观察。阳性生长须经涂片染色验证。培养至8周未见细菌生长，为结核分枝杆菌培养阴性。
3.结核杆菌的抗酸染色
取上述结核分枝杆菌培养阳性菌苔少许，均匀涂布于玻片上，火焰上固定，用石碳酸复红液染色，火焰上加热5分钟，3％盐酸酒精脱色1分钟左右，用水冲洗，再用碱性美兰溶液复染1分钟，水冲洗，吸干水分后，油镜下观察，染色为红色者为结核分枝杆菌。
《实施例3》可利霉素抗结核分支杆菌活性实验
1.绝对浓度法
1)抗结核药物
可利霉素标准品：源自中国药品生物制品检定所；对照药：异烟肼、利福平采用Sigma公司的标准品。抗结核药物按一定浓度配制成母液，然后按一定量加入培养基配制成所须剂量(表2)。
2)菌株接种
临床标本分离的菌株，涂片确认为抗酸菌的培养物，用含10％吐温80的生理盐水稀释，与McFarland标准比浊管(广东环凯微生物科技有限公司)比浊，制备10-2mg/mL菌液，接种于含检测药物的培养基。
每批试验设阴性和阳性对照。阴性对照为未加药物培养基。阳性对照为结核分枝杆菌标准株H37Rv。每管接种量为0.1mL。接种后的培养基置37℃培养。
表2抗结核药物加入培养基内的剂量
Figure PCTCN2016108502-appb-000002
注：本发明实验中的一线药物用量是根据临床实际用量设计的。
3)结果观察和判定
菌液接种后第3日观察一次，以后每周观察一次，4周报告结果。
培养结果按下列标准判定：
①培养8周未见菌落生长，报告分枝杆菌培养阴性(-)；
②培养基斜面菌落生长20个以下，报告分枝杆菌阳性与菌落数；
③培养基斜面菌落分散生长，菌落数20个以上，占据斜面面积1/4以下者，报告分枝杆菌阳性(1+)；
④培养基斜面菌落分散生长，占据斜面1/4以上，1/2以下者，报告分枝杆菌培养阳性(2+)；
⑤培养基斜面菌落密集生长或部分融合，占据斜面1/2以上，3/4以下者，报告分枝杆菌阳性(3+)；
⑥培养基斜面菌落密集生长，占据斜面面积3/4以上者，报告分枝杆菌阳性(4+)。
2.可利霉素抗结核分支杆菌活性
1)对可利霉素敏感的临床结核分支杆菌
240株临床分离鉴定的结核分支杆菌中，对可利霉素1或20(μg/ml)敏感的菌株有172株(表3)。由表3可见，可利霉素对临床分离的172株敏感菌株中，有92株菌的敏感度与实验室质控株H37Rv相当，占有效菌株的53.48％；其他80株的敏感度明显高于实验室质控株；有54株对1μg/ml的可利霉素敏感，占总有效菌的31.39％。说明可利霉素对临床分离的结核分支杆菌有较好的活性。
表3对可利霉素敏感的临床结核分支杆菌
Figure PCTCN2016108502-appb-000003
Figure PCTCN2016108502-appb-000004
Figure PCTCN2016108502-appb-000005
质控：用结核分支杆菌实验室标准株H37Rv做为阳性对照；
对照：不加药培养基。所有分离菌株在无药培养基生长均为4+。
2)可利霉素与异烟肼对某些临床结核分支杆菌活性的比较
将可利霉素与临床一线抗结核药物异烟肼对临床结核分支杆菌活性进行比较，结果表明，可利霉素20μg/ml对37株异烟肼耐药的临床结核分支杆菌有效(表4)。
表4可利霉素与异烟肼对临床结核分支杆菌灵敏度的比较
Figure PCTCN2016108502-appb-000006
Figure PCTCN2016108502-appb-000007
3)可利霉素与利福平对某些临床结核分支杆菌活性的比较
将可利霉素与临床一线抗结核药物利福平对临床分离的结核分支杆菌活性进行比较，实验结果表明，可利霉素20μg/ml对39株利福平耐药的临床结核分支杆菌有效(表5)。
表5可利霉素与利福平对临床结核分支杆菌灵敏度的比较
Figure PCTCN2016108502-appb-000008
Figure PCTCN2016108502-appb-000009
可利霉素对某些临床结核分支杆菌活性优于250μg/ml的利福平。
4)可利霉素对某些耐异烟肼和耐利福平的临床结核分支杆菌活性比较
将可利霉素与耐异烟肼和耐利福平的临床结核分支杆菌活性进行比较，实验结果表明，可利霉素20μg/ml对23株临床结核分支杆菌活性优于1μg/ml异烟肼和250μg/ml利福平(表6)。
表6可利霉素对某些耐异烟肼和耐利福平的临床结核分支杆菌活性比较
Figure PCTCN2016108502-appb-000010
本发明实验研究结果表明，可利霉素不仅对临床一线抗结核药物异烟肼和利福平敏感菌有活性，而且对部分异烟肼和利福平的耐药菌也有活性，有望在临床治疗耐药结核分枝杆菌感染疾病中得到新的应用。

Claims (6)

  1. 可利霉素在制备抗结核分枝杆菌感染药物中的应用。
  2. 以可利霉素为有效成分与药学上可接受的载体组成的组合物在制备抗结核分枝杆菌感染药物中的应用。
  3. 权利要求1或2所述的应用，其特征是，抗结核分枝杆菌感染治疗采用的可利霉素或可利霉素的组合物制备成口服剂型、注射剂型或其它任何适宜的剂型。
  4. 一种抗结核分枝杆菌感染的方法，包括给予患者有效量的可利霉素。
  5. 一种抗结核分枝杆菌感染的方法，包括给予患者有效量的可利霉素与药学上可接受的载体组成的组合物。
  6. 权利要求4或5所述的方法，其特征是，抗结核分枝杆菌感染治疗采用的可利霉素或可利霉素的组合物制备成口服剂型、注射剂型或其它任何适宜的剂型。
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