CN114432428A - PGLa提高细菌对抗生素的敏感性和延缓细菌耐药性产生的应用 - Google Patents
PGLa提高细菌对抗生素的敏感性和延缓细菌耐药性产生的应用 Download PDFInfo
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- CN114432428A CN114432428A CN202210176576.8A CN202210176576A CN114432428A CN 114432428 A CN114432428 A CN 114432428A CN 202210176576 A CN202210176576 A CN 202210176576A CN 114432428 A CN114432428 A CN 114432428A
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Abstract
本发明公开了PGLa提高细菌对抗生素的敏感性和延缓细菌耐药性产生的应用。通过多肽PGLa和抗生素联合用药,提高耐药菌对抗生素的敏感性,并延缓细菌耐药性产生,在克服药物应用中的细菌耐药性上具有更好的效果以及更高的安全性和操作性。
Description
技术领域
本发明属于医药技术领域,涉及多肽PGLa与抗生素的联合用药,以及多肽PGLa在提高细菌对抗生素敏感性、延缓细菌耐药性产生方面的应用。
背景技术
抗生素耐药菌严重危害人类健康。美国疾病控制和预防中心披露,美国每年约有200多万人遭受耐药菌感染,至少造成2.3万人死亡。世界卫生组织预测,到2050年耐药菌感染导致的死亡可能达到每年1000万人。在畜牧养殖中已全面实行“饲料端禁抗,养殖端限抗”。因此急需开发新型抗菌药物,挽救抗生素疗效。
抗生素联合用药是挽救抗生素药效的有效方案,例如利奈唑胺与亚抑制浓度的亚胺培南联用,可以对耐甲氧西林金黄色葡萄球菌发挥杀菌作用,磷霉素与阿米卡星联用,可以对多重耐药的鲍曼不动杆菌发挥杀菌作用。但抗生素联合并没有使抗生素的用量绝对减少。非抗生素类抗菌物质往往需要很大剂量才能在与抗生素联用后发挥作用,导致毒副作用较大。因此急需开发能与抗生素联合用药的非抗生素类抗菌物质。
PGLa是一种由非洲爪蟾皮肤颗粒腺体分泌的阳离子多肽,由21个氨基酸残基组成,分子量约为2.0KDa。PGLa富含赖氨酸,因此带有正电荷,而细菌细胞膜上脂多糖、肽聚糖等分子带负电,因此PGLa通过静电吸附与细菌细胞膜相互作用并通过形成α螺旋插入在细菌细胞膜上,当PGLa堆积达到细菌细胞膜承受的阈值浓度,会导致细胞膜形成孔洞,细胞内容物释放进而引起细菌裂解死亡。
中国专利CN1046462公开了生物活性两亲肽与抗菌素(例如妥布霉素、庆大霉素、红霉素、青霉素、单菌霉素、林可霉素、氯霉素、万古霉素、卡那霉素等抗生素)联用具有增效作用,即所述生物活性两亲肽能够有效的增加抗菌素的抗靶细胞(例如大肠杆菌、绿脓假单胞菌、金黄色酿脓葡萄菌的非耐药菌株)的活性,并使预防、破坏、抑制靶细胞生长所需的抗菌素用量(例如采用20%的MIC)及其自身用量减少。但单独使用PGLa或抗生素时,均需要较高浓度才能抑制耐药菌的生长,二者联用对抑制耐药菌的增效作用尚难以确定。
发明内容
针对以上现有技术中存在的问题,本发明提供了PGLa提高细菌对抗生素的敏感性和延缓细菌耐药性产生的应用,达到安全、有效抑制细菌(特别是耐药菌)的目的。
为达到上述目的,本发明采用了以下技术方案:
一种用于抑制细菌生长和杀死细菌的联合抗菌药物,该药物包括抗生素以及多肽,其中多肽与抗生素的重量比为1:(0.01-100),多肽为PGLa或PGLa的类似物。
优选的,所述PGLa的类似物是将PGLa的氨基酸序列(GMASKAGAIAGKIAKVALKAL)中位于氨基端第2位的氨基酸Met替换为Ala、Gly、Ser、Thr中的任意一种氨基酸。
优选的,所述抗生素选自氨苄青霉素(氨苄西林)、萘夫西林、羧苄西林、苯唑西林、青霉素G、头孢唑林、头孢氨苄、头孢西丁、头孢噻肟、头孢呋辛、头孢他啶、氯霉素、左氧氟沙星、环丙沙星、四环素、万古霉素、亚胺培南、克林霉素、达托霉素、卡那霉素、庆大霉素、利福平、红霉素、巴洛沙星、金霉素等中的一种或多种。其中,氨苄青霉素等为青霉素类抗生素,头孢他啶等为头孢菌素类抗生素,氯霉素等为氯霉素类抗生素,红霉素等为大环内酯类抗生素,左氧氟沙星、环丙沙星、巴洛沙星等为喹诺酮类抗生素,四环素等为四环素类抗生素,亚胺培南等为碳青霉烯类抗生素,达托霉素等为环脂肽类抗生素,卡那霉素和庆大霉素等为氨基糖苷类抗生素,克林霉素等为林可酰胺类抗生素,万古霉素等为糖肽类抗生素。
优选的,所述药物中多肽与头孢菌素类抗生素的重量比为1:(0.12-2),例如PGLa与头孢唑林的重量比为1:(1-2),PGLa与头孢噻肟的重量比为1:(0.125-0.25);多肽与喹诺酮类抗生素的重量比为1:(0.12-0.25),例如PGLa与环丙沙星的重量比为1:(0.125-0.25);多肽与碳青霉烯类抗生素的重量比为1:(0.03-10),例如PGLa与亚胺培南的重量比为1:(0.03125-0.167);多肽与环脂肽类抗生素的重量比为1:(2-4),例如PGLa与达托霉素的重量比为1:(2-4);多肽与青霉素类抗生素的重量比为1:(0.12-2),例如PGLa与氨苄西林的重量比为1:(1-2),PGLa与萘夫西林的重量比为1:(0.5-1),PGLa与羧苄青霉素(羧苄西林)的重量比为1:(0.5-1),PGLa与苯唑西林的重量比为1:(0.125-0.5);多肽与氨基糖苷类抗生素的重量比为1:(1-4),例如PGLa与卡那霉素的重量比为1:(1-4);多肽与氯霉素类抗生素的重量比为1:(0.2-0.5),例如PGLa与氯霉素的重量比为1:(0.25-0.5)。
优选的,所述药物为口服剂或注射剂。
优选的,所述药物还包括辅料,所述辅料选自药学上可接受的载体、赋形剂、溶剂中的一种或多种。
优选的,所述药学上可接受的载体选自填充剂、崩解剂、润湿剂、pH调节剂、抗氧化剂等中的一种或多种。
优选的,所述填充剂选自淀粉类、微晶纤维素、乳糖、甘露醇中的一种或多种。
优选的,所述崩解剂选自羟丙基淀粉、羧甲基淀粉钠、羧甲基淀粉钠、交联羧甲基纤维素钠、交联聚维酮、低取代羟丙基纤维素、玉米淀粉中的一种或多种。
优选的,所述润湿剂选自水、乙醇水溶液中的一种。
优选的,所述pH调节剂选自氢氧化钠、盐酸、磷酸、磷酸盐、柠檬酸、柠檬酸盐、甘氨酸、赖氨酸中的一种或多种,pH调节剂调节pH值在5.5-8.5之间(例如6.5-7.5)。
优选的,所述抗氧剂选自亚硫酸钠、亚硫酸氢钠、焦亚硫酸钠中的一种或多种。
优选的,所述溶剂为注射用水。
优选的,所述多肽可人工合成。
上述联合抗菌药物是在多肽与抗生素联合应用情况下依据相应细菌敏感性实验的发现而提出的。实验中PGLa的使用浓度为0.1-512mg/L时,能够提高细菌(特别是耐药菌株)对抗生素的敏感性;PGLa的使用浓度为0.1-512mg/L时,还能够延缓细菌抗生素耐药性的发生。而实验中抗生素的使用浓度可降至2mg/L。
优选的,所述细菌为抗生素敏感菌或抗生素耐药菌,具体包括大肠杆菌(大肠埃希菌)、溶藻弧菌、铜绿假单胞菌等革兰氏阴性菌和表皮葡萄球菌、金黄色葡萄球菌、链球菌、蜡样芽胞杆菌、粪肠球菌、枯草芽孢杆菌、破伤风杆菌等革兰氏阳性菌。
优选的,所述多肽的用量为0.01μg-1g/次/kg体重,抗生素按照用量为0.1mg-1mg/次/kg体重与多肽联用。
本发明的有益效果体现在:
本发明的联合抗菌药物的主要成分是多肽(例如PGLa)和抗生素,二者按一定比例组合,使得所述药物可以作为抑菌剂、杀菌剂并通过提高耐药菌对抗生素的敏感性、延缓细菌耐药性产生,解决细菌耐药性导致的抗生素治疗效果不佳的问题,且安全性和操作性更高。
进一步的,本发明通过将PGLa与特定类别的抗生素联用,可以将抗生素(例如头孢菌素类等)对耐药菌的最低抑菌浓度降低到敏感阈值以下,恢复细菌(特别是耐药菌)对抗生素的敏感性,并延缓细菌抗生素(例如青霉素类等)耐药性的产生(即PGLa还可以作为延缓细菌对抗生素敏感性的制剂主要成分)。
附图说明
图1为金黄色葡萄球菌耐药菌株对4μg/mL头孢呋辛(添加不同浓度多肽PGLa)的敏感性。
图2为金黄色葡萄球菌耐药菌株对其他类抗生素(添加32μg/mL多肽PGLa)的敏感性。
图3为铜绿假单胞菌对10μg/mL亚胺培南(添加不同浓度多肽PGLa)的敏感性。
图4为多种细菌及其耐药菌株对16μg/mL头孢呋辛(添加32μg/mL多肽PGLa)的敏感性。
具体实施方式
下面结合附图和实施例对本发明作进一步详细说明。应理解,以下实施例仅用于解释本发明,而不用于限制本发明的保护范围。
(一)金黄色葡萄球菌的耐药性测定
金黄色葡萄球菌是常见的食源性致病菌,广泛存在于自然环境中。金黄色葡萄球菌在适当的条件下能够产生肠毒素,引起食物中毒。
测定金黄色葡萄球菌对多种抗生素的最小抑菌浓度的实验步骤如下:
1.菌株选择:实验选取了金黄色葡萄球菌Mu50,Mu50是从病患上分离的一株多重耐药型金黄色葡萄球菌。
2.抗菌药物的配制:
氯霉素、四环素使用乙醇配制,氯霉素母液浓度为10mg/mL,四环素母液浓度为5mg/mL;其余抗生素均用无菌无离子水配制,其中氨苄西林、萘夫西林、羧苄西林、苯唑西林、头孢唑林、头孢氨苄、头孢西丁、头孢噻肟、头孢呋辛、头孢他啶、卡那霉素、庆大霉素、克林霉素的母液浓度为50mg/mL,左氧氟沙星、环丙沙星、万古霉素、亚胺培南、达托霉素的母液浓度为10mg/mL。
3.测定最小抑菌浓度(MIC)
按照美国临床和实验室标准化协会(CLSI)文件M100中的实验指导方案,使用微量肉汤稀释法测定抗菌药物的MIC,所有测试均在阳离子调节的Mueller Hinton肉汤(CAMHB)中进行。在96孔板每孔中加入100μL二倍梯度稀释的抗菌药物稀释液,将细菌浓度调至0.5麦氏浊度(约1.5×108CFU/mL),再稀释150倍,然后添加到每个孔中(每孔100μL),使得细菌终浓度约为5×105CFU/mL,含有细菌悬液但未添加抗菌药物的孔用作阳性对照,仅添加CAMHIB培养基的孔用作阴性对照。将96孔板至于恒温孵育箱中,于35℃孵育20-24小时后用酶标仪测量。大肠杆菌Escherichia coli ATCC 25922用作质控菌株。
MIC定义为抑制90%以上细菌生长的最低抑菌浓度,肉眼可观察到孔中溶液澄清。每个浓度重复3个孔,至少进行3次独立测量。MIC测定结果见表1,根据CLSI判定药物敏感性的判断点。
抑制率=(1-(OD600nm处理组-OD600nm阴性对照组)/(OD600nm阳性对照组-OD600nm阴性对照组))×100%
表1.金黄色葡萄球菌Mu50的耐药性
测定结果(表1)显示,Mu50对万古霉素为敏感(≤2μg/mL),而对其余所测药物均为耐药。据此得出Mu50对青霉素类、头孢菌素类一代、头孢菌素类二代、头孢菌素类三代、氯霉素类、林可酰胺类、喹诺酮类、四环素类、碳青霉烯类、环脂肽类、氨基糖苷类抗生素均耐药。
按照上述方法,测定了PGLa对Mu50的MIC的值为128μg/mL。
(二)多肽PGLa提高金黄色葡萄球菌耐药菌株对头孢呋辛的敏感性
为了研究多肽PGLa是否有助于头孢菌素类抗生素对金黄色葡萄球菌耐药菌株的杀菌作用,在抗生素头孢呋辛中添加不同浓度的多肽PGLa并进行了实验。
首先,配制4μg/mL的头孢呋辛溶液8份,再分别添加终浓度为64μg/mL、32μg/mL、16μg/mL、8μg/mL、4μg/mL、2μg/mL、1μg/mL的多肽PGLa(PGLa与头孢呋辛的体积比均为1:1),充分溶解均匀后分别等体积加入按照(一)中操作制备好的7份菌液中,剩余1份头孢呋辛溶液等体积加入按照(一)中操作制备好的第8份菌液中,置于37℃恒温振荡器200rpm培养3h后,分别吸取100μL新鲜菌液涂布于提前准备好的LB营养琼脂培养基上,37℃培养24h后进行菌落计数得活菌数,计算杀菌率,分别参照未添加多肽PGLa(0μg/mL)情况下(即经4μg/mL头孢呋辛处理3小时)的杀菌率计算杀菌倍数。
结果如图1所示,在添加多肽PGLa后头孢呋辛对金黄色葡萄球菌Mu50的杀菌率提高,而且随着多肽PGLa含量的提高,杀菌率也随之提高。具体的实验数据如下:多肽PGLa添加量是4μg/mL时,杀菌率提高了40倍;多肽PGLa添加量是8μg/mL时,杀菌率提高了90倍;多肽PGLa添加量是16μg/mL时,杀菌率提高了151倍;多肽PGLa添加量是32μg/mL时,杀菌率提高了285倍。多肽PGLa添加量是64μg/mL时,杀菌率仍为285倍,杀菌效率提高不再明显。
(三)多肽PGLa提高金黄色葡萄球菌耐药菌株对其他类抗生素的敏感性
为了研究添加一定比例的多肽PGLa后,除了头孢菌素类(例如头孢呋辛)以外的其他β-内酰胺类及非β-内酰胺类的抗生素对耐药菌是否有效,进行了下面的实验。
按照(一)中操作制备金黄色葡萄球菌耐药菌株菌液,向菌液中等体积加入抗生素和多肽(菌液:抗生素:多肽体积比=2:1:1),使得抗生素终浓度分别为:32μg/mL氨苄青霉素、64μg/mL卡那霉素、8μg/mL氯霉素、1μg/mL亚胺培南、32μg/mL头孢唑林、16μg/mL萘夫西林、4μg/mL环丙沙星、64μg/mL达托霉素等,并与添加至终浓度为32μg/mL的多肽PGLa混合(PGLa与抗生素的体积比均为1:1),处理4h后吸取新鲜菌液,经转接LB营养琼脂培养基培养后统计活菌数,计算杀菌率。
结果如图2所示,添加多肽PGLa后,抗生素对金黄色葡萄球菌Mu50的杀菌率最少提高了11倍,最大提高了125倍。并且当添加PGLa终浓度至32μg/mL时,氨苄西林(终浓度为32μg/mL)、达托霉素(终浓度为64μg/mL)、氯霉素(终浓度为8μg/mL)、头孢唑林(32μg/mL),及卡那霉素(终浓度为64μg/mL)的杀菌率,与环丙沙星及亚胺培南相比,提高不明显。
(四)多肽PGLa提高铜绿假单胞菌对碳青霉烯类抗生素的敏感性
为了研究多肽PGLa是否有助于碳青霉烯类抗生素对临床分离铜绿假单胞菌(耐药菌株,亚胺培南对其MIC值为128μg/mL)的杀菌作用,在抗生素亚胺培南中添加不同浓度的多肽PGLa并进行了实验。
首先,配制10μg/mL的亚胺培南溶液8份,分别与浓度为0μg/mL、1μg/mL、2μg/mL、4μg/mL、8μg/mL、16μg/mL、32μg/mL、64μg/mL的多肽PGLa,按体积比为1:1充分溶解均匀后,分别加入按照(一)中操作制备好的8份铜绿假单胞菌的菌液中,置于37℃恒温振荡器200rpm培养3h后,分别吸取100μL新鲜菌液涂布于提前准备好的LB营养琼脂培养基上,37℃培养24h后进行菌落计数得活菌数,计算杀菌率,分别参照未添加多肽PGLa(0μg/mL)情况下的杀菌率(即经10μg/mL亚胺培南处理3小时)计算杀菌倍数。
结果如图3所示,在添加多肽PGLa后亚胺培南对铜绿假单胞菌的杀菌率提高,而且随着多肽PGLa含量的提高,杀菌率也随之提高。具体的实验数据如下:PGLa添加量是2μg/mL时,杀菌率提高了5倍,PGLa添加量是4μg/mL时,杀菌率提高了12倍,PGLa添加量是8μg/mL时,杀菌率提高了41倍,PGLa添加量是32μg/mL时,杀菌率提高了245倍。当PGLa添加量是64μg/mL时(PGLa与亚胺培南比例为32:5),杀菌率提高了245倍,杀菌率较PGLa添加量为32μg/mL时提高不再明显。
(五)多肽PGLa提高多种细菌及其耐药菌株对头孢菌素类抗生素的敏感性
为研究添加多肽PGLa后,表皮葡萄球菌BB2321、表皮葡萄球菌耐药菌株BB2632、大肠杆菌K88、鸡源大肠埃希菌耐药菌和铜绿假单胞菌ATCC27853及其临床分离耐药菌株对头孢菌素类抗生素的敏感性是否提高,按照(一)中操作分别制备以上3种细菌的敏感菌株、耐药菌株的菌液,分别向各菌液中等体积加入由16μg/mL的头孢呋辛与添加至终浓度为32μg/mL的多肽PGLa构成的混合液(菌液:PGLa:头孢呋辛体积比=2:1:1),处理4h后吸取新鲜菌液,经转接LB营养琼脂培养基培养后,计算杀菌率。
结果如图4所示,添加多肽PGLa后,对3种细菌的敏感菌株的杀菌率提高了约2倍,对其相应的耐药菌株的杀菌率提高了7-24倍。
综上所述,在抗生素中添加一定比例的多肽PGLa,可以提高金黄色葡萄球菌等细菌及其耐药菌株对头孢呋辛等抗生素的敏感性,多肽PGLa与抗生素联用后,多肽PGLa的工作浓度位于0.1-512mg/L之间,而抗生素的工作浓度最多可由1024mg/L下降至2mg/L,且联用后抑菌效果显著。以上各实验结果表明,可以通过抗生素与多肽PGLa联用,提高细菌对抗生素敏感性,以此治疗细菌及其耐药菌株引起的症状。
(六)多肽PGLa延缓金黄色葡萄球菌对抗生素耐药性的产生
实验分三组:第一组将金黄色葡萄球菌ATCC29213连续在1/2MIC值浓度的苯唑西林、氯霉素、卡那霉素下连续传20代,所添加的三种抗生素终浓度分别为1μg/mL、2μg/mL、8μg/mL;第二组在第一组每次传代的基础上再添加8μg/mL的多肽PGLa,第三组连续传代金黄色葡萄球菌ATCC29213,不添加抗生素及多肽PGLa。
传代完成后,分别挑取最终第20代的金黄色葡萄球菌ATCC29213,测定其对苯唑西林、氯霉素、卡那霉素的MIC值。
实验数据(表2)表明,添加一定浓度的PGLa可以延缓金黄色葡萄球菌对抗生素耐药性的产生。
表2.延缓耐药性实验结果
(七)小鼠实验
取实验用小鼠200只,体重稳定在100g左右,其中100只全部静脉注射新鲜Mu50菌液0.2mL,另外100只全部静脉注射敏感型金黄色葡萄球菌ATCC29213,耐药金黄色葡萄球菌与敏感金黄色葡萄球菌均平均分配为5组,每组20只小鼠,在注射菌液后间隔3h后开始实验。第一组,注射4μg/mL的头孢呋辛(剂量是0.2mg/次/kg)和生理盐水的混合溶液;第二组,注射4μg/mL的头孢呋辛和0.01mg/次/kg的多肽PGLa的混合溶液(该溶液中PGLa的浓度是1μg/mL);第三组,注射4μg/mL的头孢呋辛和0.1mg/次/kg的多肽PGLa的混合溶液(该溶液中PGLa的浓度是10μg/mL);第四组,注射4μg/mL的头孢呋辛和1mg/次/Kg的多肽PGLa的混合溶液(该溶液中PGLa的浓度是100μg/mL);第五组,注射4μg/mL的头孢呋辛和10mg/次/Kg的多肽PGLa的混合溶液(该溶液中PGLa的浓度是1000μg/mL)。
同等条件下饲养,观察各组小鼠在注射Mu50耐药菌和ATCC29213敏感菌一周内的死亡率,死亡率数据统计见表3、表4。
结果表明,注射Mu50组中从第二组开始随所添加PGLa浓度越高小鼠死亡率越低,第四组小鼠死亡率为0,因此添加PGLa能显著提高小鼠存活率,说明了PGLa与抗生素联用在小鼠体内抑制并杀死金黄色葡萄菌的效果更好。并且,注射ATCC29213组中第二组及第三组的小鼠死亡率比注射Mu50组中第二组及第三组的死亡率更小,表明同等剂量下,敏感菌ATCC29213比耐药菌Mu50更容易在小鼠体内被杀死。
表3.小鼠死亡率-Mu50
表4.小鼠死亡率-ATCC29213
(八)其他实验的结果
添加1mg/mL多肽PGLa后,金黄色葡萄球菌Mu50对卡那霉素、庆大霉素、头孢他啶、头孢噻肟、亚胺培南、环丙沙星、氯霉素的敏感性提高了2-128倍。
在8μg/mL的氨苄青霉素中添加终浓度16μg/mL的多肽PGLa后,对金黄色葡萄球菌、大肠杆菌和铜绿假单胞菌的杀菌率提高了5-10倍,对其相应的耐药菌株的杀菌率提高了8-12倍。并且可以延缓金黄色葡萄球菌对氨苄青霉素(以及卡那霉素和庆大霉素)耐药性的发生。
总之,本发明通过将抗生素与多肽PGLa的联用,能够明显提高抗生素对耐药菌的抑制能力,以及明显延缓细菌耐药性的发生,为克服细菌耐药性提供了操作性、安全性高的方案。
Claims (10)
1.一种抗菌药物组合物,其特征在于:该组合物包括抗生素及多肽,所述组合物中多肽与抗生素的重量比为1:(0.01-100),多肽为PGLa或PGLa的类似物。
2.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述抗生素选自青霉素类、头孢菌素类、氯霉素类、大环内酯类、喹诺酮类、四环素类、碳青霉烯类、环脂肽类、氨基糖苷类、林可酰胺类、糖肽类抗生素中的一种或多种。
3.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述组合物中多肽与头孢菌素类抗生素的重量比为1:(0.12-2)。
4.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述组合物中多肽与喹诺酮类抗生素的重量比为1:(0.12-0.25)。
5.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述组合物中多肽与碳青霉烯类抗生素的重量比为1:(0.03-10)。
6.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述组合物中多肽与环脂肽类抗生素的重量比为1:(2-4)。
7.根据权利要求1所述一种抗菌药物组合物,其特征在于:所述组合物中,多肽与青霉素类抗生素的重量比为1:(0.12-2),或多肽与氨基糖苷类抗生素的重量比为1:(1-4),或多肽与氯霉素类抗生素的重量比为1:(0.2-0.5)。
8.多肽在制备用于提高细菌对抗生素的敏感性和/或延缓细菌耐药性的发生的药物中的应用,其特征在于:所述多肽与抗生素的重量比为1:(0.01-100),多肽为PGLa或PGLa的类似物。
9.根据权利要求8所述的应用,其特征在于:所述药物按照多肽的用量为0.01μg-1g/次/kg体重给药,抗生素按照用量为0.1mg-1mg/次/kg与该药物联用。
10.根据权利要求8所述的应用,其特征在于:所述细菌为抗生素敏感菌或抗生素耐药菌。
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