CN107847465B - 抑制具抗药性微生物的医药组合物及其应用 - Google Patents

抑制具抗药性微生物的医药组合物及其应用 Download PDF

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CN107847465B
CN107847465B CN201680045423.2A CN201680045423A CN107847465B CN 107847465 B CN107847465 B CN 107847465B CN 201680045423 A CN201680045423 A CN 201680045423A CN 107847465 B CN107847465 B CN 107847465B
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陈丘泓
陈晔
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Peptide Cham Biotech Co ltd
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    • 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
    • A61K31/21—Esters, e.g. nitroglycerine, selenocyanates
    • A61K31/27—Esters, e.g. nitroglycerine, selenocyanates of carbamic or thiocarbamic acids, meprobamate, carbachol, neostigmine
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    • 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
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

一种抑制具抗药性微生物的抗生素，所述抗生素为Fmoc‑Ala‑OPfp，其对人体细胞不具毒性且同时具有能有效抑制具抗药性微生物的功效。

Description

抑制具抗药性微生物的医药组合物及其应用
技术领域
本发明是有关于一种抑制具有抗药性的微生物的医药组合物及其应用。
背景技术
抗药性(drug resistance)是指药物的治疗疾病或改善病人征状的效力降低。当投入药物浓度不足而不能杀死或抑制病原时，残留的细菌可能具有抵抗此种药物的能力。例如细菌可能因抗生素产生的活性氧诱发DNA突变而造成耐药性。
细菌对抗生素产生抗药性的机制基本上是由基因控制，这些基因统称为抗药性基因。这些抗药性基因中，有些是某些细菌天生就具有的，也可能是经由质体(plasmid)或跳跃子(transposon)传递而从其他细菌得到的。在有抗生素的环境中，自然会将药物可杀死的细菌个体淘汰掉，而只剩下带有抗药性基因的细菌，因此在有抗生素的环境下，就会衍生出越来越多的抗药性细菌。这就是为何抗生素使用越多的地区，抗药性细菌的比例越高的主要原因。
近年来因缺乏治疗感染抗药性细菌的抗生素及医界过度滥用，造成强抗药性的突变种细菌，导致欧美地区每年超过4万名丧生。因此，面对俗称「超级细菌」的抗药性细菌，各国皆着手研发新型抗生素。
安比西林(Ampicillin)作为临床医学第一线抗生素，为医疗院所及畜产业常态性使用，也因此抗生素的滥用，出现越来越多具抗药性的细菌。其中，台湾已有71％门诊患者体内大肠杆菌对于安比西林产生抗药性，而美国则为39.3％。该状况不仅出现在第一线抗生素药物，于大部分抗生素均无效治疗后所使用的万古霉素(Vancomycin)，亦由于抗生素滥用的情况下同样产生了具有抗药性的细菌，如抗万古霉素肠球菌(Vancomycin-Resistant Enterococcus，VRE)。
然而，过去药厂为节省成本，因此使用较为简易的研发方法，仅修饰现有抗生素的结构，例如：置换官能基，但经些许时间使用后具抗药性的细菌仍能对修饰后的抗生素产生新的抗药性，系故若再无新型抗生素，将不具有可有效治疗细菌的抗生素而造成确诊细菌感染的病患无药可救的情况。
发明内容
有鉴于此，本发明揭示了一种抗生素并可抑制具抗药性的微生物，所述抗生素为Fmoc-Ala-OPfp。
本发明所述抑制微生物生长的医药组合物是一种抗生素，芴甲氧羰基-L-丙氨酸五氟苯酯(N-(9-fluorenylmethoxycarbonyl)-L-alanine pentafluorophenyl，即Fmoc-Ala-OPfp，CAS:86060-86-8)，所述医药组合物为一种丙氨酸(alanine，Ala)衍生物，其结构式如式I所示。
Figure GDA0001567107670000021
较佳地，前述具抗药性的微生物是选自葛兰氏阳性菌、金黄色葡萄球菌Staphylococcus aureus、表皮葡萄球菌Staphylococcus epidermidis、肺炎链球菌Streptococcus Pneumoniae、肠球菌Enterococci、破伤风梭菌Clostridium tetani肉毒杆菌Clostridium botulinum中的一种或多种。
较佳地，前述具抗药性的微生物为金黄色葡萄球菌Staphylococcus aureus。
较佳地，前述医药组合物(即抗生素)的有效浓度为2.5-100uM。
较佳地，前述医药组合物(即抗生素)的有效浓度为2.5uM。
较佳地，前述抗药性为对万古霉素药物具耐受性。
本发明更近一步提供一种可抑制微生物生长的医药组合物，所述医药组合物包含Fmoc-Ala-OPfp。
本发明更近一步提供一种抑制微生物生长的医药组合物的应用，所述医药组合物包含Fmoc-Ala-OPfp。
附图说明
图1为本发明医药组合物Fmoc-Ala-OPfp的溶血性测试。
图2为本发明医药组合物Fmoc-Ala-OPfp的人类细胞的毒性测试。
图3为本发明医药组合物Fmoc-Ala-OPfp与广效性抗生素安比西林、万古霉素的抑制抗药性的黄金葡萄球菌的抑制效果比较图。
具体实施方式
实施例1.抗菌性测试
参见图3，本发明进行细菌培养试验，分别于培养基中加入特定浓度的本发明医药组合物Fmoc-Ala-OPfp、安比西林、万古霉素作为实验组，以及再另一培养基中不添加任何药剂作为控制组，并接种相同菌量的金黄色葡萄球菌后于37℃培养箱中培养；经由测定不同时间点的菌液浓度(OD630)以了解Fmoc-Ala-OPfp、安比西林、万古霉素的抑菌效果。
其中，在培养基中仅添加少量的Fmoc-Ala-OPfp(1μg/ml)，即使在接种细菌7小时后，仍可有效发挥等同于5.3μg/ml的安比西林及2.13μg/ml万古霉素的的抗菌效果，证明Fmoc-Ala-OPfp能有效抑制细菌的生长。
实施例2.溶血性测试
参见图1，新鲜人类红血球细胞由0.9％的生理食盐水冲洗3次后回溶于2％(V/V)的生理食盐水之中，将50μL的红血球悬浮液及50μL的本发明医药组合物(10、50及100μg/ml抗生素Fmoc-Ala-OPfp)或Triton X-100加入96格盘，其中红血球最后的浓度为2％(V/V)。于培养箱中于37℃并辅以轻微摇动的条件下培养1小时，样本以2000g离心30分钟。取50μL样本的上清液并滴入新的96格盘并以O.D.值490nm测量其吸光值(OD490)，磷酸缓冲溶液(PBS)生理食盐水及Triton X-100分别为阴性控制组及阳性控制组。
实施例3.细胞培养
参见图2，L929及MEF细胞株为哺乳类动细胞株并于Dulbecco's modified Eagle培养液(DMEM培养基)中培养，其中培养液加入100μg/mL streptomycin(链霉素)、100U/mLpenicillin(青霉素)以及10％(v/v)fetal bovine serum(胎牛血清)，接着将细胞株培于含有5％CO2且温度为37℃的潮湿培养箱中。
如图2所示，本发明抗生素Fmoc-Ala-OPfp对哺乳类动细胞株具有极低之细胞毒性。
实施例4.存活率分析
通过MTT分析法测试化合物对哺乳类细胞所造成的毒性，将细胞培养于灭菌后的96格盘并以100μL培养液培养24小时，接着化合物稀释于培养液之中，并继续培养24小时。之后将3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide(MTT)溶液加入96格盘的每一格并培养4小时。经离心后，移除上清液并加入100μL dimethyl sulfoxide(DMSO，二甲亚砜)溶解所有沉淀物。使用酵素免疫分析测读仪以570nm测量吸光值。
如图1、2所示，因此，人类红血球的溶血性实验及MTT分析测试Fmoc-Ala-OPfp的毒性，于100μg/mL的浓度下Fmoc-Ala-OPfp仍具有低溶血性，而于MTT分析中100μg/mL的浓度下Fmoc-Ala-OPfp对细胞仅具有10-20％的毒性。
上列详细说明系针对本发明之一可行实施例的具体说明，惟该实施例并非用以限制本发明的专利范围，凡未脱离本发明技艺精神所为的等效实施或变更，均应包含于本发明的专利范围中。

Claims (3)

1.一种包含有芴甲氧羰基-L-丙氨酸五氟苯酯Fmoc-Ala-Opfp的医药组合物供用于制备用来抑制金黄色葡萄球菌(Staphylococcus aureus)生长的医药品的用途，其中芴甲氧羰基-L-丙氨酸五氟苯酯的结构式如式I所示：
Figure FDA0002508726900000011
2.如权利要求1所述的用途，其特征是，所述芴甲氧羰基-L-丙氨酸五氟苯酯的有效浓度为2.5-100uM。
3.如权利要求1所述的用途，其特征是，所述芴甲氧羰基-L-丙氨酸五氟苯酯的有效浓度为2.5uM。
CN201680045423.2A 2015-08-03 2016-08-03 抑制具抗药性微生物的医药组合物及其应用 Expired - Fee Related CN107847465B (zh)

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