CN116421623A - 一种含高价银的抗菌剂及其制备方法 - Google Patents
一种含高价银的抗菌剂及其制备方法 Download PDFInfo
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- CN116421623A CN116421623A CN202310377024.8A CN202310377024A CN116421623A CN 116421623 A CN116421623 A CN 116421623A CN 202310377024 A CN202310377024 A CN 202310377024A CN 116421623 A CN116421623 A CN 116421623A
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- Prior art keywords
- antibacterial agent
- silver
- valence
- containing high
- valence silver
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/57—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone
- A61K31/573—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids substituted in position 17 beta by a chain of two carbon atoms, e.g. pregnane or progesterone substituted in position 21, e.g. cortisone, dexamethasone, prednisone or aldosterone
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- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于抗菌剂技术领域，具体涉及一种含高价银的抗菌剂及其制备方法。本发明以含量为3‑20μg/mL的Ag3+为基础，辅以炉甘石、ZnO和海藻粉，制成凝胶剂或乳剂时，具有优异的抑菌效果，且用于动物杀菌和止痒时表现优异。本发明提供的含高价银的抗菌剂具有优异的生物安全性，能抑制细菌真菌，并具有优异的伤口修复能力。本发明还提供了含高价银的抗菌剂的制备方法，通过N2和O3封闭，使得封闭后的含高价银的抗菌剂保有抑菌效果。
Description
技术领域
本发明属于抗菌剂技术领域，具体涉及一种含高价银的抗菌剂及其制备方法。
背景技术
瘙痒是兽医临床上，特别是犬猫等小动物诊疗实践中畜主最常反映的症状之一。犬猫的瘙痒症一般在临床上表现为刮擦撕咬，尤其喜欢频繁地在爪和尾部舔咬，瘙痒会阴部，摩擦脸部，在地毯和家具上磨蹭或频繁地在地上打滚。兽医人员既可通过观察患犬猫的擦痒行为，也可通过观察体表上显著的红斑、表皮的脱落、皮肤上的癣斑、体表擦伤的伤口或脱毛等临床表现认定瘙痒症状的存在。细菌感染是家养宠物瘙痒的主要病因，细菌主要有中间葡萄球菌、金黄色葡萄球菌、表皮葡萄球菌、链球菌、化脓性棒状杆菌和奇异变形杆菌等。目前主要是根据药敏实验结果指导临床用药，增强机体抵抗力，全身和局部应用抗生素。红霉素、林可霉素、克拉维酸阿莫西林、TMP、头孢菌素、百多邦、甲硝哇、利福平和恩诺沙星等药物都是常用药，但由于抗生素的耐药性以及易过敏的问题，导致需要一种新型抗菌剂用于动物杀菌和止痒。
银系抗菌剂的抗菌活性与其价态密切相关，价态越高，抗菌性能越强，研究表明Ag3+的抗菌活性是Ag2+的200倍，Ag2+的抗菌活性是Ag+的60倍。然而，目前应用的银系抗菌剂主要是以磺胺嘧啶银、硝酸银为代表的单价银抗菌剂或纳米银抗菌剂，由于高价银难以长期保存导致高价银抗菌剂的研发与应用较少。
抗生素的应用使得细菌感染的危险系数大大降低，但随着抗生素的选择作用，细菌抗性越来越强，抗生素的研发速度渐渐无法满足临床需求。银系抗菌剂因其接触杀菌与光催化杀菌作用，并不针对某一定向靶点，使其具有广谱抗菌性能具有极高的应用价值，但高价银难以长期保存导致实际应用困难。由此，如何制得一种含高价银的抗菌剂，将高价银与生物体相容性好的物质进行配合，控制银的缓释机制，增强抗菌剂的长效抗菌机制，使其稳定存在，并减少银的细胞毒性成为重要的研究课题。
发明内容
本发明的目的在于提供一种含高价银的抗菌剂，将高价银与生物体相容性好的物质进行配合，控制银的缓释机制，增强抗菌剂的长效抗菌机制，使其稳定存在，并减少银的细胞毒性。
基于上述目的，本申请通过提供一种含高价银的抗菌剂及其制备方法来解决该领域中的这种需要。
一方面，本发明涉及一种含高价银的抗菌剂，含有Ag3+3-20μg/mL。
进一步地，本发明提供的含高价银的抗菌剂中，以质量比计，包括Ag3+3-20μg/mL，炉甘石10-25mg/mL、ZnO 1-6mg/mL、海藻粉14-42mg/mL。
进一步地，本发明提供的含高价银的抗菌剂中，以质量比计，所述炉甘石中ZnO含量不低于40％。
进一步地，本发明提供的含高价银的抗菌剂中，所述海藻粉为小叶海藻粉。
进一步地，本发明提供的含高价银的抗菌剂中，制备方法包括：将所述Ag3+的配合物、炉甘石、ZnO、海藻粉分散在水中，制成水剂、凝胶剂或乳剂中的一种，调节pH至3-3.5，加压排出空气打入N2和O3封闭。
进一步地，本发明提供的含高价银的抗菌剂中，所述配合物为Ag3+-聚六亚甲基双胍配合物。
进一步地，本发明提供的含高价银的抗菌剂中，制成凝胶剂；所述凝胶剂中包含卡波姆。
进一步地，本发明提供的含高价银的抗菌剂中，制成乳剂；所述乳剂中包含硬脂酸和单甘酯。
进一步地，本发明提供的含高价银的抗菌剂中，以体积比计，所述N2和O3的配比为3-5:1。
进一步地，本发明提供的含高价银的抗菌剂中，加压排出空气打入N2和O3封闭；置于容器中，然后打入N2排空所述容器中的气体，最后打入O3加压封闭保存，所述加压压力为保持所述容器内部压力不低于0.8Mpa。
本发明与现有技术相比具有以下有益效果或者优点：
本发明提供了一种含高价银的抗菌剂，将高价银与生物体相容性好的物质进行配合，控制银的缓释机制，增强抗菌剂的长效抗菌机制，使其稳定存在，并减少银的细胞毒性。本发明还提供了该含高价银的抗菌剂的制备方法，将该抗菌剂制备成水剂、凝胶剂或乳剂，便于使用，且通过N2和O3的加压保存，能稳定该抗菌剂的抗菌性能，以及用于动物杀菌和止痒时表现优异。本发明提供了一种含高价银的抗菌剂，其具有优异的生物安全性，能抑制细菌真菌，并具有优异的伤口修复能力。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例，都属于本发明保护的范围。
下述各实施例中实验方法和检测方法，如无特殊说明，均为常规方法；所述药剂和材料，如无特殊说明，均可在市场上购买得到；所述指标数据，如无特殊说明，均为常规测量方法。
实施例1
本实施例提供了一种含高价银的抗菌剂的水剂制备试验。
(1)Ag3+-聚六亚甲基双胍配合物的制备
按照n(己二胺盐酸盐):n(双氰胺钠)＝1:2.1的摩尔比取己二胺盐酸盐和双氰胺钠溶解在正丁醇中，在溶剂沸腾条件下，反应8h，蒸出溶剂正丁醇，并将所得白色固体1，6-双氰基胍基已烷用水与无水乙醇反复洗涤至熔点在210℃以上。
将所得1，6-双氰基胍基己烷与己二胺盐酸盐按照等摩尔比加入到四口烧瓶中，加入少量水，并用稀盐酸调节pH值在6.8左右，使其充分混合。在55℃下，真空除水，待水完全除去用N2气保护并升温至155～160℃反应60～180min，每隔30min，取样1次，得到平均相对分子质量在4406的PHMB。
将PHMB溶解在去离子水中，配成质量分数20％的水溶液备用。在250mL三口烧瓶中，加入76mL去离子水，加入880μL 20％的PHMB水溶液，并搅拌均匀。在避光条件下，先后慢慢向其中滴加0.04mol/L的AgNO3 10mL和0.08mol/L的K2S2O8 12mL，用pH＝14的NaOH溶液调节pH值在11～12之间，反应16h。反应完毕后将离心所得橙棕色固体，用去离子水洗去可溶性杂质，然后在45℃下干燥。
(2)含高价银的抗菌剂，剂型为水剂
炉甘石购自睿诚康医药科技公司，ZnO含量不低于40％；小叶海藻粉购自宁夏香草生物技术有限公司，为马尾藻科植物羊栖菜Sargassum fusiforme(Harv.)Setch.的干燥藻体。
将Ag3+-聚六亚甲基双胍配合物、炉甘石、ZnO、小叶海藻粉分散在水中，水浴加热至25-35℃，加入苯甲酸甲酯，搅拌溶解后，调节pH至3-3.5制得含高价银的抗菌剂的水剂，加压排出空气打入N2和O3封闭。
其中，Ag3+-聚六亚甲基双胍配合物3μg/mL、炉甘石10mg/mL、ZnO 1mg/mL、小叶海藻粉14mg/mL、苯甲酸甲酯35mg/mL。
以体积比计，N2和O3的配比为5:1。
以体积比计，N2和O3的体积和为含高价银的抗菌剂体积的1/10。
实施例2
本实施例提供了一种含高价银的抗菌剂的凝胶剂制备试验。
本实施例的制备原料来源同实施例1，制备方法如下：
将卡波姆分散在部分水中预溶，水浴加热至25-35℃保持，加入苯甲酸甲酯、Ag3+-聚六亚甲基双胍配合物、炉甘石、ZnO、小叶海藻粉，补足剩余水，调节pH至3-3.5制得含高价银的抗菌剂的凝胶剂，加压排出空气打入N2和O3封闭。
其中，Ag3+10μg/mL、炉甘石15mg/mL、ZnO 3mg/mL、小叶海藻粉20mg/mL，卡波姆10mg/mL、苯甲酸甲酯35mg/mL。以体积比计，N2和O3的配比为4:1。以体积比计，N2和O3的体积和为含高价银的抗菌剂体积的1/10。
实施例3
本实施例提供了一种含高价银的抗菌剂的乳剂制备试验。
本实施例的制备原料来源同实施例1，制备方法如下：
将卡波姆溶解在部分水中，充分溶胀24小时，制成流状A液备用。将硬脂酸和单甘酯加入部分水中，加热到80℃，充分溶解得B液。将Ag3+-聚六亚甲基双胍配合物、炉甘石、ZnO、小叶海藻粉分散在剩余水中，水浴加热至25-35℃，加入苯甲酸甲酯，搅拌溶解后与A液、B液混合搅拌，然后加入植物油，调节pH至3-3.5，超声乳化，即为高价银的抗菌剂的乳剂，加压排出空气打入N2和O3封闭。
其中，Ag3+20μg/mL、炉甘石25mg/mL、ZnO 6mg/mL、小叶海藻粉42mg/mL、硬脂酸200mg/mL、单甘酯50mg/mL、苯甲酸甲酯35mg/mL。以质量比计，卡波姆的用量为5％，植物油的用量为10％。以体积比计，N2和O3的配比为3:1。以体积比计，N2和O3的体积和为含高价银的抗菌剂体积的1/10。
实施例4
本实施例提供了一种含高价银的抗菌剂的动物安全性试验。
试验动物：新西兰家兔，体重2-2.5kg，雌雄各半。
新西兰家兔20只，体重2.0～2.5kg，雌雄各半，均分为5组，每组4只。第1组为空白对照组，第2组为实施例1破损皮肤给药组(破损皮肤用针尖划破，以出血为度)，第3组为实施例2破损皮肤给药组，第4组为实施例3破损皮肤给药组，第5组为实施例1完整皮肤给药组。
于给药前24h，将家免背部脊柱两侧毛用8％硫化钠溶液脱去，面积约为150cm2。实验时脱毛区给药组分别涂抹实施例1-3提供的含高价银的抗菌剂中的一种8g，对照组脱毛区涂抹清水作为对照，然后用一层油纸及二层纱布覆盖，再以无刺激胶布封闭、固定，动物分笼饲养。所有动物给受试物24h后，用温开水洗去残留的受试物，去除受试物后，于1周内，每日观察并记录动物的体重、皮肤、毛发、眼和黏膜的变化、呼吸、循环、中枢神经系统、四肢活动等的变化，若有动物死亡，则需进行尸检和肉眼观察。当有肉眼可见病变时，则需进行病理组织学检查。
实验结果表明，在整个观察期间，所有动物正常生长，毛色光亮，行为活动、饮水、饮食及大小便均正常，口、鼻、耳等处无异常分泌物，呼吸、循环和中枢神经系统均正常。所有动物均存活，且给药组和对照组动物未见明显差异。提示本品对家兔皮肤用药无明显的急性毒性反应。
实施例5
本实施例提供了一种含高价银的抗菌剂的体外抗菌试验。
供试液：实施例1-3提供的含高价银的抗菌剂。
测定最低抑菌浓度(MIC)和最低杀菌浓度(MBC)：用琼脂二倍稀释法测定MIC。于含有供试液的系列琼脂平皿表面点种细菌，菌液最终浓度为105CFU/mL，需氧菌37℃培养18h，真菌27℃培养24h，厌氧菌置厌氧孵箱37℃孵育48h，未见细菌生长的平皿内所含供试药的最低浓度为其最小抑菌浓度(MIC，mg/L)。实验中用金黄色葡萄球菌，大肠埃希氏菌作质控菌株。用肉汤二倍稀释法测MIC，再取未见细菌生长管培养液0.1mL于无药琼脂平皿表面，用无菌玻璃棒推匀，培养(条件同MIC测定)，仍无细菌生长的最低药物浓度为其最低杀菌浓度(MBC)，结果见表1。
表1，抗菌试验结果
如表1所示，本发明提供的含高价银的抗菌剂金黄色葡萄球菌等9个试验菌种具有明显的抑菌或杀菌作用。由此，本发明提供的含高价银的抗菌剂具有优异的抑菌效果，且适用于动物杀菌和止痒。
进一步，本发明提供了实施例2含高价银的抗菌剂凝胶剂形封闭7天后的抗菌试验。
样品处理：实施例2提供的一种含高价银的抗菌剂的凝胶剂封闭7天后称取5g，放无菌研钵中研磨、粉碎、混匀，放入无菌平血中真接进行试验。
试验菌株：大肠杆菌(ATCC 8099)，培养9代、白色念珠菌(ATCC 10231)，培养9代、金黄色葡萄球菌(ATCC 6538)，培养9代，绿脓杆菌(ATCC 9027)培养9代，由中国菌种保藏中心提供。每组试验重复5次，试验温度为20±2℃，实施例2提供的一种含高价银的抗菌剂的凝胶剂杀菌效果如表2所示。
表2，高价银的抗菌剂凝胶剂形的杀菌效果
注[1]：阳性对照组平均活菌数(平均3.1×104cfu/片，2.8×104cfu/片～3.6×104cFu/片)，阴性对照组无菌生长；
注[2]：阳性对照组平均活菌数(平均2.7×104cfu/片，2.2×104cfu/片～2.9×104cFu/片)，阴性对照组无菌生长；
注[3]：阳性对照组平均活菌数(平均2.5×104cfu/片，2.0×104cfu/片～2.6×104cFu/片)，阴性对照组无菌生长；
注[4]：阳性对照组平均活菌数(平均2.1×104cfu/片，1.9×104cfu/片～2.6×104cFu/片)，阴性对照组无菌生长。
由表2可知，在试验温度为20±2℃的条件下，实施例2提供的一种含高价银的抗菌剂的凝胶剂封闭7天后作用5min，对大肠杆菌(ATCC 8099)、白色念珠菌(ATCC 10231)、金黄色葡萄球菌(ATCC 6538)，绿脓杆菌(ATCC 9027)的平均杀菌率可达90％以上。
实施例6
本实施例提供了一种含高价银的抗菌剂的促进伤口愈合试验。
试验动物：新西兰家兔，体重2-2.5kg，雌雄各半。
供试液：1#实施例1提供的含高价银的抗菌剂封闭7天，2#实施例2提供的含高价银的抗菌剂封闭7天，3#实施例3提供的含高价银的抗菌剂封闭7天，4#实施例1提供的含高价银的抗菌剂不含炉甘石、ZnO和小叶海藻粉封闭7天，5#实施例2提供的含高价银的抗菌剂不含炉甘石、ZnO和小叶海藻粉封闭7天，6#实施例3提供的含高价银的抗菌剂不含炉甘石、ZnO和小叶海藻粉封闭7天，7#凡士林(阴性对照组)，8#京万红软膏(阳性对照组)。
于试验前一天剃掉家兔背部两侧的背毛，去毛范围约为3cm×3cm。注射戊巴比妥钠(30mg/mL)对家兔进行麻醉，后用碘酒清洗背部去毛区域皮肤，用灭菌刀片按20cm×15cm大小划破家兔表皮(注意皮肤破损仅达表皮，不要伤及真皮)，去除表皮形成2.0cm×1.5cm的去皮破损伤口。用生理盐水清洗伤口，并在该破损皮区涂抹对应组别的药物，用2层纱布敷贴在涂药后的伤口处，然后用一层无刺激塑料膜覆盖，再用无刺激胶布固定。给家兔上药前用生理盐水清洗伤口，每天在同一时间点给药一次，连续给药14d，给药量为5g/d。在每次给药前观察家兔伤口的愈合情况，并观察家兔给药后的精神状态、活动等。分别在给药的第7天和第14天对家兔的伤口进行拍照，观察伤口的愈合情况：是否有出血、渗透液、化脓和结痂等现象，并用量尺测量伤口愈合的大小尺寸，计算伤口愈合率，试验结果如表。
表3，高价银的抗菌剂凝胶剂形的杀菌效果
如表3所示，本发明提供的一种含高价银的抗菌剂具有优异的伤口修复能力，相较未加入炉甘石、ZnO和小叶海藻粉，其伤口修复能力得到显著提升。
如上所述，即可较好地实现本发明，上述的实施例仅仅是对本发明的优选实施方式进行描述，并非对本发明的范围进行限定，在不脱离本发明设计精神的前提下，本领域普通技术人员对本发明的技术方案做出的各种改变和改进，均应落入本发明确定的保护范围内。
Claims (10)
1.一种含高价银的抗菌剂，其特征在于，含有Ag3+3-20μg/mL。
2.根据权利要求1所述的含高价银的抗菌剂，其特征在于，以质量比计，包括Ag3+3-20μg/mL，炉甘石10-25mg/mL、ZnO 1-6mg/mL、海藻粉14-42mg/mL。
3.根据权利要求2所述的含高价银的抗菌剂，其特征在于，以质量比计，所述炉甘石中ZnO含量不低于40％。
4.根据权利要求2所述的含高价银的抗菌剂，其特征在于，所述海藻粉为小叶海藻粉。
5.根据权利要求2所述的含高价银的抗菌剂，其特征在于，制备方法包括：将所述Ag3+的配合物、炉甘石、ZnO、海藻粉分散在水中，制成水剂、凝胶剂或乳剂中的一种，调节pH至3-3.5，加压排出空气打入N2和O3封闭。
6.根据权利要求5所述的含高价银的抗菌剂，其特征在于，所述配合物为Ag3+-聚六亚甲基双胍配合物。
7.根据权利要求5所述的含高价银的抗菌剂，其特征在于，制成凝胶剂；所述凝胶剂中包含卡波姆。
8.根据权利要求5所述的含高价银的抗菌剂，其特征在于，制成乳剂；所述乳剂中包含硬脂酸和单甘酯。
9.根据权利要求5所述的含高价银的抗菌剂，其特征在于，以体积比计，所述N2和O3的配比为3-5:1。
10.根据权利要求5所述的含高价银的抗菌剂，其特征在于，加压排出空气打入N2和O3封闭；置于容器中，然后打入N2排空所述容器中的气体，最后打入O3加压封闭保存，所述加压压力为保持所述容器内部压力不低于0.8Mpa。
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