CN116785288A - 千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用 - Google Patents
千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用 Download PDFInfo
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Abstract
本发明公开了千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用。千金藤素与盐酸特比耐芬联合应用时，对耐药白色念珠菌表现出显著的协同作用，对白色念珠菌的抑菌效果显著。
Description
技术领域
本发明涉及医药技术领域，具体涉及千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用。
背景技术
这里的陈述仅提供与本发明相关的背景技术，而不必然地构成现有技术。
真菌感染的患病率及病死率呈持续上升趋势，而且耐药形势堪忧，常见的病原真菌包括白色念珠菌、曲霉等。
白色念珠菌又名白假丝酵母菌，是自然界广泛存在的一种真菌，其属于机会致病菌，也是临床上分离最多的真菌。当人体免疫力下降或菌群失调时，白色念珠菌可引起感染，如鹅口疮、肺炎、肠炎、支气管炎、肾盂肾炎、脑膜炎、心内膜炎等。
白色念珠菌的治疗药物一般包括外用药物、口服药物和静脉注射药物，外用药物包括硝酸咪康唑栓、酮康唑乳膏、环吡酮胺乳膏；口服药物包括伊曲康唑胶囊、盐酸特比萘芬片、氟康唑胶囊等；静脉注射药物包括氟康唑氯化钠注射液、氟康唑葡萄糖注射液、注射用伏立康唑等。
近年来，随着AIDS患者的不断增多、高效广谱抗生素和皮质类固醇激素的广泛应用、抗肿瘤治疗的深入开展以及器官移植、介入治疗的不断实施，白念珠菌感染的发生率呈上升趋势。虽然已有多种抗真菌药物如咪唑类、多烯类、烯丙胺类等可以有效防治念珠菌病；但随着抗真菌药物的广泛应用，分离到的耐药白色念珠菌逐渐增多，现有的抗真菌药物对该类耐药白色念珠菌的治疗效果较差，急需寻找克服耐药白色念珠菌的新药手段。
发明内容
针对现有技术存在的不足，本发明的目的是提供千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用。
为了实现上述目的，本发明是通过如下的技术方案来实现：
第一方面，本发明提供千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用。
千金藤最初被用于中药，为防己科植物千金藤Stephania japonica(Thunb.)Miers的根。在1914年被植物学家Bunzo Hayata报道，并在二十年后由东京大学药学家Heisaburo Kondo教授提纯并命名为千金藤素。千金藤素化学式为C37H38N2O6，白色结晶。
千金藤素与现有抗真菌药物盐酸特比萘芬联用，具有协同抗耐药白色念珠菌的作用，意义重大。
在一些实施例中，所述耐药白色念珠菌为多重耐药白色念珠菌，对咪唑类、多烯类、烯丙胺类抗真菌药均高度耐药。
在一些实施例中，千金藤素与盐酸特比萘芬联合应用时的浓度配比为2-8：16-32。
在一些实施例中，千金藤素与盐酸特比萘芬联合应用时，千金藤素的最小抑菌浓度为2μg/mL，盐酸特比萘芬的最小抑菌浓度为16μg/mL。
第三方面，本发明提供一种抗耐药白色念珠菌药物，其包括千金藤素与盐酸特比萘芬的药物组合物。
在一些实施例中，所述药物还包括药学上可接受的载体或辅料，所述载体或辅料选自水合剂、增稠剂、乳化剂、防腐剂、稳定剂、离子交换剂、助流剂、粘合剂、着色剂、香味剂、甜味剂、沉淀抑制剂、润滑剂、分散剂、稀释剂、矫味剂、抗氧化剂、等渗剂、助悬剂、乳化加速剂、缓冲剂、崩解剂、表面活性剂、吸收剂、脱模剂、涂布剂中的至少一种。
上述本发明的一种或多种实施例取得的有益效果如下：
经过试验发现，千金藤素与盐酸特比萘芬联合应用，对耐药白色念珠菌表现出显著的协同作用，对白色念珠菌的抑菌效果显著。
附图说明
构成本发明的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。
图1是千金藤素与盐酸特比萘芬联用抗耐药白色念珠菌的菌丝对比图，其中的标尺为50μm。
具体实施方式
应该指出，以下详细说明都是例示性的，旨在对本发明提供进一步的说明。除非另有指明，本发明使用的所有技术和科学术语具有与本发明所属技术领域的普通技术人员通常理解的相同含义。
下面结合实施例对本发明作进一步说明。
实施例1
实验过程和方法
药物原液的制备
将千金藤素粉末溶解于DMSO中，使其浓度为20480μg/mL，储存在灭菌的柱状药敏管中，标记后于-20℃冰箱保存备用。将盐酸特比萘芬原料药粉末溶解于无水乙醇中，使其浓度为20480μg/mL，储存在灭菌的柱状药敏管中，标记后于-20℃冰箱保存备用。
菌株的准备
使用一次性接种环将存储的白色念珠菌耐药菌株划线接种于YPD固体培养基上，35℃培养48h，并传代3次以完全活化菌株。挑取上述培养基中的单个菌落重悬于YPD液体培养基中，于35℃恒温培养过夜。使用麦氏比浊管调整菌悬液为2×106个/mL，备用。
药物联合作用测定
本实施例使用无菌的96孔板，按照美国临床和实验室标准协会CLSI(M27)进行标准的操作。先将不同梯度浓度的两个药物工作液分别加入96孔板的孔中，每孔50μL(盐酸特比萘芬终浓度：1μg/mL、2μg/mL、4μg/mL、8μg/mL、16μg/mL、32μg/mL、64μg/mL、128μg/mL、256μg/mL、512μg/mL，千金藤素终浓度：16μg/mL、32μg/mL、64μg/mL、128μg/mL、256μg/mL、512μg/mL)；再将制备好的标准菌悬液(终浓度：103个/mL)加入含药液的96孔板的孔中，每孔100μL。生长对照孔中加入100μL菌悬液和100μL培养基，最后一列孔为阴性对照孔(空白对照)，仅有培养基。药敏板其余孔用培养基补齐至200μL，配置完成后置于恒温培养箱内，35℃培养24小时后观察结果。
实验结果判读
参照CLSI标准，判读时与生长对照孔进行对比，在自然光线下肉眼判读各孔中菌的生长，MIC定义为能够抑制80％以上的药物浓度。
协同作用评价
在实验室中，以部分抑菌浓度指数(fractional inhibitory concentrationindex,FICI)作为联合药敏试验效果的判断依据：FICI＝FICA+FICB＝MICA联用÷MICA单用+MICB联用÷MICB单用。式中：MICA和MICB是A、B药物单独用药时的MIC值；CA和CB是A、B两药联用时各自MIC值。根据FICI的范围可以判断联合药敏试验的效果：FICI≤0.5为协同作用。
表1千金藤素与盐酸特比萘芬联用抗6株耐药白色念珠菌的作用
A：盐酸特比萘芬；B：千金藤素。
抑制菌丝实验
过夜培养的真菌细胞，于RPMI1640培养基中调整菌浓度至1×105cells/mL，加入不同浓度的药物，第一组为对照组，对照组不加入药物；第二组中加入1μg/ml盐酸特比萘芬；第三组中加入16μg/ml千金藤素；第四组中加入1μg/ml盐酸特比萘芬+16μg/ml千金藤素。
37℃静置培养3h(1)、6h(2)和24h(3)后，使用显微镜观察菌丝形态，如图1所示。
实验结论
千金藤素与盐酸特比萘芬联用可协同抑制耐药型白色念珠菌，两者表现为明显的协同作用。另外，千金藤素与盐酸特比萘芬联用可显著抑制耐药型白色念珠菌的菌丝这一重要的毒力因子。
以上所述仅为本发明的优选实施例而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
Claims (6)
1.千金藤素与盐酸特比萘芬联用在制备耐药白色念珠菌药物中的应用。
2.根据权利要求1所述的应用，其特征在于：所述耐药白色念珠菌为多重耐药白色念珠菌。
3.根据权利要求1所述的应用，其特征在于：千金藤素与盐酸特比萘芬联合应用时的浓度配比为2-8：16-32。
4.根据权利要求1所述的应用，其特征在于：千金藤素与盐酸特比萘芬联合应用时，千金藤素的最小抑菌浓度为2μg/mL，盐酸特比萘芬的最小抑菌浓度为16μg/mL。
5.一种抗耐药白色念珠菌药物，其特征在于：其包括千金藤素与盐酸特比萘芬的组合物。
6.根据权利要求5所述的抗耐药白色念珠菌药物，其特征在于：所述药物还包括药学上可接受的载体或辅料，所述载体或辅料选自水合剂、增稠剂、乳化剂、防腐剂、稳定剂、离子交换剂、助流剂、粘合剂、着色剂、香味剂、甜味剂、沉淀抑制剂、润滑剂、分散剂、稀释剂、矫味剂、抗氧化剂、等渗剂、助悬剂、乳化加速剂、缓冲剂、崩解剂、表面活性剂、吸收剂、脱模剂、涂布剂中的至少一种。
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