CN107835691A - 抵抗碳青霉烯耐药性阴性细菌的含有作为活性成分的adk蛋白的抗菌组合物 - Google Patents
抵抗碳青霉烯耐药性阴性细菌的含有作为活性成分的adk蛋白的抗菌组合物 Download PDFInfo
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- CN107835691A CN107835691A CN201680020565.3A CN201680020565A CN107835691A CN 107835691 A CN107835691 A CN 107835691A CN 201680020565 A CN201680020565 A CN 201680020565A CN 107835691 A CN107835691 A CN 107835691A
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Abstract
本发明涉及抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物,其包含作为活性成分的来源于结核分枝杆菌的腺苷酸激酶(ADK)蛋白。根据本发明的来源于结核分枝杆菌的ADK蛋白对碳青霉烯耐药性革兰氏阴性细菌具有优异的抗微生物活性,因此可作为抗菌组合物用于各种领域。
Description
技术领域
[1]本发明涉及抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物,其包含作为活性成分的来源于结核分枝杆菌(Mycobacterium tuberculosis)的腺苷酸激酶(ADK)蛋白。
背景技术
[2]通常,抗菌剂统称为抗微生物剂,特别是具有抗微生物作用的物质,特别是通过抑制系统(在该系统内,细菌合成细胞壁、蛋白等)而具有优异的抗微生物效果的物质,或由这些物质制备的剂。抗菌剂的主要成分主要从真菌中提取,并且这些抗菌剂如今被广泛用于治疗由细菌感染等引起的疾病。
[3]从弗莱明(Fleming)在20世纪发现青霉素抗生素开始,已经研发了许多抗微生物剂和抗生素以避免细菌感染引起的疾病。这种抗菌剂在我们的生活中占据重要的地位,广泛用于各种应用,如药物、食品、化妆品防腐剂等中。然而,在使用化学合成材料的抗菌剂的情况下,对其具有耐药性的细菌数量逐渐增加,因此其用途越来越受限制。
[4]抗微生物剂耐药性细菌是指对某些抗微生物剂具有耐药性并因此对其不响应的细菌。例如,对青霉素不响应的青霉素耐药性金黄色葡萄球菌(Staphylococcus aureus)属于这些细菌。另外,学术界在1961年首次报道甲氧苯青霉素耐药性金黄色葡萄球菌(MRSA)并且自那时以来其已经成为全球主要的致病性感染性细菌,并且于1988年首先在欧洲发现对万古霉素(vancomycin)有耐药性的万古霉素耐药性肠球菌(VRE)是公知的,并且在90年代后期,在日本、美国、法国和韩国报道了万古霉素中等耐药性金黄色葡萄球菌(VISA)。另外,在2002年由疾病控制中心(the Centers for Disease Control)首次在全球报道万古霉素耐药性金黄色葡萄球菌(VRSA),其中万古霉素作为属于最常见的人类感染致病菌的金黄色葡萄球菌的最后治疗剂是公知的,并且因此所谓超级细菌的增殖的可能性大大增加。
[5]同时,在抗生素领域被视为最重要的β-内酰胺抗生素包含通常称为青霉素的青霉烷抗生素(penam antibiotics)、通常称为头孢菌素的头孢菌素抗生素(cefemantibiotics)、青霉烯抗生素(penem antibiotics)和碳青霉烯类抗生素(carbapenemantibiotics)。其中,碳青霉烯类抗生素的实例包含可商购的亚胺培南(imipenem)、帕尼培南(panipenem)、美罗培南(meropenem)、厄他培南(ertapenem)等。
[6]因此,本发明的发明人努力研发新的抗微生物剂,结果证实了来源于结核分枝杆菌(Mycobacterium tuberculosis)的腺苷酸激酶或腺苷激酶(ADK)蛋白对碳青霉烯耐药性革兰氏阴性细菌表现出优异的抗微生物活性,从而完成了本发明。
发明内容
【技术问题】
[7]本发明的目的是提供一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物,其包含作为活性成分的ADK蛋白。
[8]本发明的另一个目的是提供一种用于预防或治疗由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的组合物,该组合物包含作为活性成分的ADK蛋白。
【技术方案】
[9]本发明提供抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物、准药物、食品添加剂或饲料添加剂,其包含作为活性成分的ADK蛋白。
[10]本发明还提供了用于预防或治疗由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的药物组合物或食品组合物,所述组合物包含作为活性成分的ADK蛋白。
【有益效果】
[11]根据本发明,来源于结核分枝杆菌(Mycobacterium tuberculosis)的ADK蛋白对碳青霉烯耐药性革兰氏阴性细菌具有优异的抗菌活性,并且因此可作为抗菌组合物用于各种领域。
附图说明
[12]图1是显示重组ADK蛋白的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析结果的图像。
具体实施方式
[13]在下文中,将更详细地描述本发明。
[14]本发明提供了一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物,其包含作为活性成分的ADK蛋白。
[15]本文所用的术语“抗菌”或“抗菌活性”是指抵抗诸如细菌或真菌之类的微生物的性质,更具体地,是指抗生素等抑制细菌生长或增殖的性质等。
[16]本文所用的术语“抗菌组合物”是指具有抑制微生物(如细菌或真菌)的生长和发育的活性的组合物,并且可以包含在需要抗微生物作用的各种领域中使用的所有形式,例如药物、准药物、食品添加剂、饲料添加剂等。特别地,抗菌组合物可以用于与微生物直接相关的产品中,例如医疗领域中的抗生素或污染抑制剂;用于防腐或抗微生物目的的食品;在农业中用于抗微生物、杀菌或消毒目的;化妆品或日常用品,以抑制头皮屑、防治运动员脚,或抑制腋窝的气味,或用于抗痤疮的目的;或用于防腐、抗微生物或杀菌目的的清洁用洗涤剂、洗盘子用洗涤剂等,但本发明不限于上述目的。
[17]本发明的ADK蛋白来源于结核分枝杆菌(Mycobacterium tuberculosis),可以优选具有由SEQ ID NO:1表示的氨基酸序列,可以通过由SEQ ID NO:2所表示的碱基序列编码,并且包含ADK蛋白的功能等同物。本文所用的术语“功能等同物”是指由于氨基酸的添加、取代或缺失而与氨基酸序列SEQ ID NO:1具有至少70%,优选至少80%,更优选至少90%,最优选至少95%的序列同源性的蛋白,并且该蛋白表现出与具有氨基酸序列SEQ IDNO:1的蛋白或由碱基序列SEQ ID NO:2编码的蛋白的生理活性基本上相同的生理活性。
[18]本发明的ADK蛋白包含具有其天然氨基酸序列的蛋白和其氨基酸序列变体的蛋白。ADK蛋白的变体是指通过ADK天然氨基酸序列和一个或多个氨基酸残基或其组合的缺失、插入或非保守或保守取代具有不同序列的蛋白。在蛋白和肽中并不总体上改变分子的活性的氨基酸交换是本技术领域已知的。ADK蛋白或其变体可以天然提取或合成(Merrifield,J.Amer.Chem.Soc.85:2149-2156,1963),或者可以通过基于DNA序列的基因重组来制备(Sambrook et al,Molecular Cloning,Cold Spring Harbor LaboratoryPress,New York,USA,2nd Edition,1989)。
[19]本发明的ADK蛋白可以由碱基序列SEQ ID NO:2编码,并且能够在功能上与核苷酸执行相同的作用的变体包含在本发明的范围内。
[20]本文所用的术语“革兰氏阴性细菌”是指通过革兰氏染色染成红色的细菌,并且通常,这些细菌对颜料和表面活性剂具有很强的耐性。本发明的革兰氏阴性菌包括含有内毒素的所有类型的革兰氏阴性细菌,其实例包含但不限于属于下述属的细菌:埃希氏杆菌(Escherichia)属、假单胞菌(Pseudomonas)属、不动杆菌(Acinetobacter)属、沙门氏菌(Salmonella)属、克雷伯氏菌(Klebsiella)属、奈瑟氏菌(Neisseria)属、肠杆菌(Enterobacter)属、志贺氏菌(Shigella)属、莫拉氏菌(Moraxella)属、幽门螺杆菌(Helicobacter)属、寡养单胞菌(Stenotrophomonas)属、扁豆属(Bdellovibrio)属和军团菌(Legionella)属。特别地,这些革兰氏阴性细菌的实例包含但不限于大肠杆菌(Escherichia coli)、绿脓假单胞菌(Pseudomonas aeruginosa)、荧光假单胞菌(Pseudomonas fluorescens)、恶臭假单胞菌(Pseudomonas putida)、绿脓假单胞菌(Pseudomonas chlororaphis)、穿孔素假单胞菌(Pseudomonas pertucinogena)、斯氏假单胞菌(Pseudomonas stutzeri)、丁香假单胞菌(Pseudomonas syringae)、鲍曼不动杆菌(Acinetobacter baumannii)、鲁氏不动杆菌(Acinetobacter lwoffii)、醋酸钙不动杆菌(Acinetobacter calcoaceticus)、溶血性不动杆菌(Acinetobacter haemolyticus)、肠炎沙门氏菌(Salmonella enterica)、邦戈沙门氏菌(Salmonella bongori)、肠炎沙门氏菌(Salmonella enteritidis)、鼠伤寒沙门氏菌(Salmonella typhimurium)、鸡沙门氏菌(Salmonella gallinarum)、鸡白痢沙门氏菌(Salmonella pullorum)、姆班达卡沙门氏菌(Salmonella mbandaka)、猪霍乱沙门氏菌(Salmonella choleraesuls)、汤姆森沙门氏菌(Salmonella thompson)、婴幼儿沙门氏菌(Salmonella infantis)、德氏沙门氏菌(Salmonella derby)、肺炎克雷伯氏菌(Klebsiella pneumonia)、肉芽肿克雷伯氏菌(Klebsiella granulomatis)、产酸克雷伯氏菌(Klebsiella oxytoca)、土生克雷伯氏菌(Klebsiella terrigena)、淋病奈瑟氏球菌(Neisseria gonorrhoeae)、脑膜炎奈瑟氏球菌(Neisseria meningitidis)、产气肠杆菌(Enterobacter aerogenes)、阴沟肠杆菌(Enterobacter cloacae)、鲍氏志贺氏菌(Shigella boydii)、痢疾志贺氏菌(Shigelladysenteriae)、福氏志贺氏菌(Shigella flexneri)、宋氏志贺氏菌(Shigella sonnei)、卡他莫拉菌(Moraxella catarrhalis)、腔隙莫拉氏菌(Moraxella lacunata)、牛莫拉氏菌(Moraxella bovis)、幽门螺杆菌(Helicobacter pylori)、海尔曼螺杆菌(Helicobacterheilmannii)、猫胃螺杆菌(Helicobacter felis)、雪貂螺杆菌(Helicobacter mustelae)、芬纳尔螺杆菌(Helicobacter fenelliae)、芜菁螺杆菌(Helicobacter rappini)、肝螺杆菌(Helicobacter hepaticus)、胆汁螺杆菌(Helicobacter bilis)、肠胃炎螺杆菌(Helicobacter pullorum)、嗜麦芽窄食单胞菌(Stenotrophomonas maltophilia)、嗜麦芽寡养单胞菌(Stenotrophomonas nitritireducens)、食菌蛭弧菌(Bdellovibriobacteriovorus)、嗜肺军团菌(Legionella pneumophila)、茴香军团菌(Legionellaanisa)、伯利恒军团菌(Legionella birminghamensis)、博兹曼军团菌(Legionellabozemanii)、辛辛那提军团菌(Legionella cincinnatiensis)、杜莫夫军团菌(Legionelladumoffii)、弗氏军团菌(Legionella feeleii)、戈氏军团菌(Legionella gormanii)、假鹤虱军团菌(Legionella hackeliae)、以色列军团菌(Legionella israelensis)、约旦军团菌(Legionella jordanis)、兰斯格军团菌(Legionella lansingensis)、长滩军团菌(Legionella longbeachae)、马氏军团菌(Legionella maceachernii)、米克达德军团菌(Legionella micdadei)、橡岭军团菌(Legionella oakridgensis)、圣海伦军团菌(Legionella sainthelensi)、图森军团菌(Legionella tucsonensis)和沃斯沃军团菌(Legionella wadsworthii)。
[21]本文所用的术语“碳青霉烯”是指具有由下式1表示的结构的一种β-内酰胺抗生素:
<式1>
其中,在式1中,R1和R2各自独立地为H、C1-C10烷基或经羟基取代的C1-C10烷基,以及
R3是H、-O-R4或-S-R4,
其中R4是经N-取代或C-取代的胺或吡咯烷。
[22]碳青霉烯类抗生素的实例包含但不限于亚胺培南(imipenem)、美罗培南(meropenem)、厄他培南(ertapenem)、多利培南(doripenem)、帕尼培南/倍他普隆(panipenem/betamipron)、比亚培南(biapenem)、阿祖培南(razupenem)、替比培南(tebipenem)、来那培南(lenapenem)、和托莫培南(tomopenem)。
[23]作为代表性碳青霉烯类抗生素的亚胺培南(imipenem)、帕尼培南(panipenem)、美罗培南(meropenem)、厄他培南(ertapenem)和多利培南(doripenem)的结构分别显示在式2、式3、式4、式5和式6中:
<式2>
<式3>
<式4>
<式5>
<式6>
[24]根据本发明的来源于结核分枝杆菌(Mycobacterium tuberculosis)的ADK蛋白对碳青霉烯耐药性革兰氏阴性细菌具有优异的抗微生物活性,且因此可在各种领域有效地用作抗菌组合物,即准药物、食品添加剂、饲料添加剂等。
[25]本发明还提供了抗碳青霉烯耐药性革兰氏阴性细菌的包含作为活性成分的ADK蛋白的抗菌准药物、抗菌食品添加剂或抗菌饲料添加剂。
[26]本发明还提供了包含作为活性成分的ADK蛋白的用于预防或治疗由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的组合物。
[27]组合物包含药物组合物或食物组合物。
[28]本发明还提供一种预防或治疗由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的方法,所述方法包含向个体施用ADK蛋白。
[29]本发明的ADK蛋白选择性地表现出对碳青霉烯耐药性革兰氏阴性细菌的优异的抗微生物活性,因此包含该ADK蛋白的组合物可有效地用于预防或治疗由碳青霉烯耐药性革兰氏阴性菌引起的感染性疾病。
[30]本文所用的术语“感染性疾病”是指由通过繁殖和侵入动物或人的导致疾病的致病微生物引起的病,致病微生物如病毒、细菌、真菌和寄生虫,并且所述术语“感染性疾病”是指根据本发明的目的的由碳青霉烯耐药性革兰氏阴性细菌引起的所有感染性疾病。例如,本发明的感染性疾病可以是呼吸系统疾病、胃肠道疾病、炎症性疾病等。特别地,传染病的实例包含但不限于肺炎、腹膜炎、脑膜炎、伤口感染、骨关节炎、胆囊炎、尿道感染、脑血管炎、心内膜炎、心肌炎、心包炎、关节炎、咽炎、淋病、杆菌性痢疾、肠炎、结膜炎、胃炎、鼓膜炎、膀胱炎、淋巴管炎和败血症。
[31]本文所用的术语“预防”是指通过施用组合物抑制感染性疾病或延迟其发作的所有行为。本文所用的术语“治疗”是指通过施用组合物减轻或有益地改变由于感染性疾病引起的症状的所有行为。
[32]本发明的药物组合物还可以包含常用的合适的载体、赋形剂或稀释剂。可以包含在本发明的药物组合物中的载体、赋形剂或稀释剂的实例包含但不限于乳糖、葡萄糖、蔗糖、山梨糖醇、甘露糖醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、金合欢胶、藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁和矿物油。
[33]本发明的药物组合物可以根据一般方法以口服或肠胃外形式施用,并且使用通常使用的稀释剂或赋形剂(如填充剂、增量剂、粘合剂、润湿剂、崩解剂、表面活性剂等)进行配制。用于口服施用的固体制剂包含片剂、丸剂、粉剂、颗粒剂、胶囊剂等,并且通过将组合物与至少一种赋形剂(如淀粉、碳酸钙、蔗糖、乳糖、明胶等)以制备这样的固体制剂。除了简单的赋形剂之外,还使用润滑剂,如硬脂酸镁和滑石粉。用于口服施用的液体制剂的实例包含悬浮液、内部使用的液体、乳剂、糖浆等,除了简单的常用稀释剂(例如水和液体石蜡)外,这些液体制剂还可以包括各种赋形剂,例如润湿剂、甜味剂、调味剂、防腐剂等。肠胃外施用的制剂包含无菌水溶液、非水溶剂、悬浮液、乳剂、冷冻干燥制剂和栓剂。非水溶剂和悬浮液的非限制性实例包含丙二醇、聚乙二醇、植物油(如橄榄油)和可注射的酯(如油酸乙酯)。栓剂基质的实例包含Witepsol、Macrogol、吐温61(Tween 61)、可可脂、月桂精、甘油胶冻(glycerogelatin)等。
[34]本文所用的术语“施用”是指通过使用适当的方法将本发明的药物组合物供应给个体。
[35]本发明的药物组合物的合适剂量可以根据患者的条件和体重、疾病严重程度、药物类型、施用途径和施用时间而变化,但可以由本领域普通技术人员适当选择。为了获得期望的效果,本发明的药物组合物可以按每天0.001mg/kg至1000mg/kg的量施用。药物组合物可以每天施用一次或多次。该剂量并不意图以任何方式限制本发明的范围。
[36]本发明的药物组合物可以通过各种途径施用给个体。可以预期所有的施用方法,例如口服注射、直肠或静脉内注射、肌肉注射、皮下注射、子宫内膜硬膜外注射和脑室内注射。然而,对于口服施用,由于蛋白被消化,因此可以配制口服组合物,使得活性成分被包被或配制成被保护免于在胃中分解。本发明的组合物可优选以注射的形式施用。
[37]根据本发明所述的药物组合物除了包含ADK蛋白之外,还可以包含一种或多种具有抗微生物活性的已知物质。
[38]在本发明中,食品组合物可优选为健康功能食品的形式。
[39]在本发明中,健康功能食品是指具有由物理、生物化学或生物技术方法提供的附加值使得相应的食品赋予或表现出适合于特定应用的预期功能的一组食品,或者被设计为使得食物的组成在身体中足以赋予关于生物防御节律控制、疾病预防和恢复等的身体调节功能的经加工的食品。
[40]健康功能食品可以包含可以使用的可食用的食品添加剂,并且还可以包含通常用于制备健康功能食品的合适的载体、赋形剂和稀释剂。
[41]当本发明的食品组合物用作食品添加剂时,该组合物可以根据常用的方法单独或与其它食品或食品成分组合使用。可以根据使用目的(预防、健康或医疗)适当地确定活性成分的混合量。通常,在食品或饮料的制备中,相对于原料,本发明的组合物可以按15重量%或以下,优选按10重量%或以下的量添加。然而,当为了健康和卫生目的或为了健康控制目的长时间摄取时,食物组合物的量可以包含在上述量的范围内或低于上述量的范围或超过上述量的范围。组合物在安全性方面没有问题,因此也可以在上述量的范围内或大于上述量的范围使用。
[42]除了上述成分外,本发明的食品组合物可以包含各种营养补充剂、维生素、电解质、调味剂、着色剂、果胶酸及其盐、海藻酸及其盐、有机酸、保护胶体增稠剂、pH调节剂、稳定剂、防腐剂、甘油、醇、碳酸饮料中使用的碳酸化剂等。此外,本发明的食品组合物可以包含用于制备天然果汁、果汁饮料和蔬菜饮料的果肉。这些成分可以单独使用或组合使用。虽然这些添加剂的比例不是非常重要,但是相对于100重量份的本发明的组合物,添加剂的量通常选自0.01重量份至0.1重量份。
[43]在下文中,将描述示例性实例来帮助理解本发明。然而,这些实施例仅用于更容易理解本发明,并不意图限制本发明的范围。
实施例1.重组ADK(Rv0733)的克隆
[44]使用结核分枝杆菌H37Rv(ATCC 27294)的基因组DNA作为模板(引物:5'-CATATGAGAGTTTTGTTGCTGGGACCG-3'(SEQ ID NO:3)和5'-AAGCTTCTTTCCCAGAGCCCGCAACGC-3'(SEQ ID NO:4))通过PCR扩增ADK(Rv0733)区。分离的PCR产物用NdeI和HindIII酶消化并插入到表达载体pET22b中。将用插入了ADK基因的载体pET22b转化的大肠杆菌BL21在LB培养基(含有100μg/ml氨苄青霉素)中于37℃培养12小时。随后,向其中加入1mM异丙基-D-硫代吡喃半乳糖苷(IPTG),然后培养6小时,用裂解缓冲液(含有1M DTT、溶菌酶和PMSF)裂解细胞。根据制造商的方法,使用镍-次氮基三乙酸琼脂糖(Ni-NTA,Invitrogen,Carlsbad,CA,,美国)纯化重组蛋白。最后,通过SDS-PAGE鉴定纯化的重组ADK蛋白。其结果示于图1中。
[45]实施例2.碳青霉烯耐药性革兰氏阴性细菌的鉴定
[46]从韩国大学(Korea University)传染病科病人血液样本分离鲍曼不动杆菌KUMC.2014.90和KUMC.2014.91菌株。为了鉴定该菌株对抗生素的耐药性,主要使用作为自动化装置的VITEK II系统,结果确认菌株对所有碳青霉烯类抗生素(即亚胺培南、美罗培南、厄他培南等)具有耐药性(确认结果值R>32)。其次,使用液体培养基微量稀释法再次证实了菌株对属于碳青霉烯类抗生素的亚胺培南和美罗培南的耐药性(确认结果值大于128μg/ml)。
[47]实验实施例1.验证ADK蛋白对碳青霉烯耐药性革兰氏阴性细菌的抗菌活性
[48]为了验证实施例1中获得的重组ADK蛋白对实施例2中获得的碳青霉烯耐药性鲍曼不动杆菌(KUMC.2014.90和KUMC.2014.91菌株)的抗菌活性,根据常规已知的方法执行Rezazurin测试以针对碳青霉烯耐药性鲍曼不动杆菌菌株测定最低抑菌浓度(MIC)和最低杀菌浓度(MBC)。作为对照,使用作为正常菌株的鲍曼不动杆菌。实验结果如下表1所示。
<表1>
[49]如表1所示,由于假设未用抗生素和ADK蛋白处理的孔的结果值为100%而计算得到50%和80%的抑制浓度,因而证实了抗生素(即氨苄青霉素、卢卡森(rukasyn)、左氧氟沙星(cravit)和谱能(prepenem))抑制属于正常菌株的鲍曼不动杆菌的生长,同时不能抑制碳青霉烯耐药性鲍曼不动杆菌(KUMC.2014.90和KUMC.2014.91菌株)的生长。相比之下,证实了ADK蛋白显著抑制了属于正常菌株的鲍曼不动杆菌属和两种类型的碳青霉烯抗药性鲍曼不动杆菌(KUMC.2014.90和KUMC.2014.91菌株)两者的生长。
[50]根据上述实验结果,证实了根据本发明,来源于结核分枝杆菌的ADK蛋白对碳青霉烯耐药性革兰氏阴性细菌具有优异的抗微生物活性。
[51]下文中,将参照以下的制造实施例对本发明的药物组合物和食品组合物进行说明。然而,这些实施例仅为了说明的目的而提供,并不意图限制本发明的范围。
[52]制备实施例1.药物组合物的制备
[53]1-1.粉末的制备
ADK蛋白 2克
乳糖 1g
将上述成分混合并将其填充到气密性包装内,从而完成粉末的制备。
[54]1-2.片剂的制备
将上述成分混合,然后根据制备片剂的一般方法制备片剂。
[55]1-3.胶囊的制备
将上述成分混合,然后根据制备胶囊的一般方法将混合物填充到明胶胶囊内,从而完成胶囊的制备。
[56]制备实施例2.食品组合物的制备
[57]2-1.保健食品的制备
[58]作为示例性实施方式,维生素混合物和矿物混合物包含用于保健食品的相对合适的成分的混合物,但混合比可以任意改变,并且根据制备保健食品的一般方法混合上述成分并制备成粒,并且可以用于根据一般方法制备保健食品组合物中。
序列表
<110> 建国大学全球本土化产学合作基金会
<120> 抵抗碳青霉烯耐药性阴性细菌的含有作为活性成分的ADK蛋白的抗菌组合物
<130> MPO17-058
<150> KR 10-2015-0047360
<151> 2015-04-03
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 181
<212> PRT
<213> 结核分枝杆菌(Mycobacterium tuberculosis)
<400> 1
Met Arg Val Leu Leu Leu Gly Pro Pro Gly Ala Gly Lys Gly Thr Gln
1 5 10 15
Ala Val Lys Leu Ala Glu Lys Leu Gly Ile Pro Gln Ile Ser Thr Gly
20 25 30
Glu Leu Phe Arg Arg Asn Ile Glu Glu Gly Thr Lys Leu Gly Val Glu
35 40 45
Ala Lys Arg Tyr Leu Asp Ala Gly Asp Leu Val Pro Ser Asp Leu Thr
50 55 60
Asn Glu Leu Val Asp Asp Arg Leu Asn Asn Pro Asp Ala Ala Asn Gly
65 70 75 80
Phe Ile Leu Asp Gly Tyr Pro Arg Ser Val Glu Gln Ala Lys Ala Leu
85 90 95
His Glu Met Leu Glu Arg Arg Gly Thr Asp Ile Asp Ala Val Leu Glu
100 105 110
Phe Arg Val Ser Glu Glu Val Leu Leu Glu Arg Leu Lys Gly Arg Gly
115 120 125
Arg Ala Asp Asp Thr Asp Asp Val Ile Leu Asn Arg Met Lys Val Tyr
130 135 140
Arg Asp Glu Thr Ala Pro Leu Leu Glu Tyr Tyr Arg Asp Gln Leu Lys
145 150 155 160
Thr Val Asp Ala Val Gly Thr Met Asp Glu Val Phe Ala Arg Ala Leu
165 170 175
Arg Ala Leu Gly Lys
180
<210> 2
<211> 546
<212> DNA
<213> 结核分枝杆菌(Mycobacterium tuberculosis)
<400> 2
gtgagagttt tgttgctggg accgcccggg gcgggcaagg ggacgcaggc ggtgaagctg 60
gccgagaagc tcgggatccc gcagatctcc accggcgaac tcttccggcg caacatcgaa 120
gagggcacca agctcggcgt ggaagccaaa cgctacttgg atgccggtga cttggtgccg 180
tccgacttga ccaatgaact cgtcgacgac cggctgaaca atccggacgc ggccaacgga 240
ttcatcttgg atggctatcc acgctcggtc gagcaggcca aggcgcttca cgagatgctc 300
gaacgccggg ggaccgacat cgacgcggtg ctggagtttc gtgtgtccga ggaggtgttg 360
ttggagcgac tcaaggggcg tggccgcgcc gacgacaccg acgacgtcat cctcaaccgg 420
atgaaggtct accgcgacga gaccgcgccg ctgctggagt actaccgcga ccaattgaag 480
accgtcgacg ccgtcggcac catggacgag gtgttcgccc gtgcgttgcg ggctctggga 540
aagtag 546
<210> 3
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221> misc_feature
<222> (1)..(27)
<223> F 引物
<400> 3
catatgagag ttttgttgct gggaccg 27
<210> 4
<211> 27
<212> DNA
<213> 人工序列(Artificial Sequence)
<220>
<221> misc_feature
<222> (1)..(27)
<223> R 引物
<400> 4
aagcttcttt cccagagccc gcaacgc 27
Claims (9)
1.一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌组合物,该抗菌组合物包含作为活性成分的腺苷酸激酶(ADK)蛋白。
2.根据权利要求1所述的抗菌组合物,其中所述ADK蛋白来源于结核分枝杆菌(Mycobacterium tuberculosis)。
3.根据权利要求1所述的抗菌组合物,其中所述ADK蛋白由SEQ ID NO:1表示。
4.根据权利要求1所述的抗菌组合物,其中所述ADK蛋白由SEQ ID NO:2所表示的碱基序列编码。
5.一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌准药物,所述抗菌准药物包含作为活性成分的腺苷酸激酶(ADK)蛋白。
6.一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌食品添加剂,该抗菌食品添加剂包含作为活性成分的腺苷酸激酶(ADK)蛋白。
7.一种抗碳青霉烯耐药性革兰氏阴性细菌的抗菌饲料添加剂,所述抗菌饲料添加剂包含作为活性成分的腺苷酸激酶(ADK)蛋白。
8.一种用于预防或治疗由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的药物组合物,所述药物组合物包含作为活性成分的腺苷酸激酶(ADK)蛋白。
9.一种用于预防或减轻由碳青霉烯耐药性革兰氏阴性细菌引起的感染性疾病的食物组合物,所述食物组合物包含作为活性成分的腺苷酸激酶(ADK)蛋白。
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PCT/KR2016/003467 WO2016159741A1 (ko) | 2015-04-03 | 2016-04-04 | Adk 단백질을 유효성분으로 포함하는 카바페넴 내성 그람음성균에 대한 항균용 조성물 |
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KR (1) | KR101995622B1 (zh) |
CN (1) | CN107835691A (zh) |
BR (1) | BR112017021225A2 (zh) |
RU (1) | RU2668160C1 (zh) |
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CN113544139A (zh) * | 2019-02-28 | 2021-10-22 | 丹迪生物科技有限公司 | 具有抗菌活性的多肽、包含该多肽的用于预防或治疗败血症的组合物和抗菌组合物 |
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KR101983668B1 (ko) * | 2017-08-22 | 2019-05-29 | 건국대학교 산학협력단 | 그람 음성 박테리아에 대한 강한 항균 활성을 갖는 벌거숭이두더지쥐 유래의 항균성 폴리펩티드 및 이를 포함하는 약학 조성물 |
KR102225240B1 (ko) * | 2019-07-03 | 2021-03-10 | 경북대학교 산학협력단 | 다제내성 병원성 세균을 효과적으로 사멸시키는 재조합 항균 단백질 Ablysin의 용도 |
KR102596822B1 (ko) * | 2020-03-20 | 2023-11-02 | 에이치엘비사이언스 주식회사 | 리포폴리사카라이드 제거용 조성물 및 키트 |
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SG11201708130PA (en) | 2017-11-29 |
US20180078619A1 (en) | 2018-03-22 |
KR101995622B1 (ko) | 2019-07-02 |
RU2668160C1 (ru) | 2018-09-26 |
BR112017021225A2 (pt) | 2018-06-26 |
EP3292765A1 (en) | 2018-03-14 |
KR20160118994A (ko) | 2016-10-12 |
JP2018516232A (ja) | 2018-06-21 |
JP6475865B2 (ja) | 2019-02-27 |
EP3292765A4 (en) | 2019-03-13 |
US10792340B2 (en) | 2020-10-06 |
WO2016159741A1 (ko) | 2016-10-06 |
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