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CN1688322B - 含有与聚果胶酸酯和edta螯合的银的抗菌溶液 - Google Patents

含有与聚果胶酸酯和edta螯合的银的抗菌溶液 Download PDF

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CN1688322B
CN1688322B CN038245620A CN03824562A CN1688322B CN 1688322 B CN1688322 B CN 1688322B CN 038245620 A CN038245620 A CN 038245620A CN 03824562 A CN03824562 A CN 03824562A CN 1688322 B CN1688322 B CN 1688322B
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埃德温·奥德尔·迈纳
克雷格·诺曼·伊托格
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Abstract

一种具有提高的长期稳定性的液体抗菌剂和清洁剂，包括至少以下主要成分：去离子水、银离子、聚果胶酸酯和乙二胺四乙酸(EDTA)。目前本技术优选实例也包括甘油，1，2-丙二醇(也就是丙二醇)；至少一种表面活性剂，其任选自烷基硫酸盐、磺酸盐、烷醇酰胺、甜菜碱、氧化胺、肌氨酸盐、和磺基琥珀酸酯；和一种缓冲化合物，其足以使pH值达到7.2到7.8范围。

Description

含有与聚果胶酸酯和EDTA螯合的银的抗菌溶液
本申请有基于临时专利申请No.60/413,379的优先权日，其申请日期为2002年9月25日。
发明领域
本申请涉及抗菌剂的制剂，特别是涉及这样的抗菌剂，其含有至少一种金属化合物作为杀灭生物的物质。
发明背景
牙科和医学专业杂志已经证明感染的蔓延，当医师和职员未能恰当清洁他们的手，感染发生在诊所、医院、病房、公共设施和其它非公共机构的设施。缺乏卫生的可怕后果经常发生，医师必须不厌其烦地清洗他们的手。酒精、过氧化氢，以及含有碘和氯的化合物已长期被认为是有效的抗微生物和抗病毒的物质，因此它们使用于擦洗和外科手术前准备是受到广泛认同的。然而，最近的研究表明，许多这些化合物的反复应用对皮肤有生理上的损害。另一方面，当微生物种群经常有规律地遭遇特定的抗微生物化合物，微生物通过发展可以排出那些化合物的化学泵逐渐进化，因此，大大地减少作为抗微生物物质的化合物的有效性。
某些金属已经显示出具有抗菌的性质。含汞的溶液，如硫柳汞和mercurachrome的酊剂，在二十世纪作为抗菌剂广泛地使用。然而，研究表明这些抗菌剂的使用导致汞通过皮肤的吸收，综合研究显示摄取甚至少量的汞导致孩子的精神发育迟缓，促进他们的发育停止。供替代抗菌化合物的研究已经广泛开展，这种抗菌化合物不会损害中枢神经系统，不会伤害皮肤和粘膜。
银是一种天然存在的金属，当局部使用时，已经显示出对众多种类致病的细菌、真菌、霉菌、寄生虫、和病毒是致死的，然而对人类身体是相对无害的。从公元前700到100年期间腓尼基人的记载显示，在长期的旅程和航海时，银容器用于保存醋、水和纯酒。同样知道，早期的美国移民者和拓荒者，将铜、金和银币使用于医疗和水净化的目的。早在19世纪年末期，银作为杀菌剂使用已得到证明。在过去的一世纪，特别是过去的二十年，银已经开始被广泛认为是一种有效的杀菌剂和杀寄生虫剂。由于这种应用，银一般用于以下形式，硝酸银、氧化银、或银化合物的胶态悬浮液。例如，硝酸银滴剂常规置于新生儿的眼睛，目的是保护新生儿的眼睛——当他们通过产道时可能已被感染。胶态银，为银元素或是氧化银微粒，悬浮在矿物油或水基质中，通过内部的摄取和局部的应用，正推广于各种医药目的。它推广用于治疗乳腺炎和治疗粘膜的感染。
没有征兆表明微生物已经或可能发展一种对银的抵抗力，因为它是从外部阻断呼吸，而不是干扰有机体的内部代谢。
最近十年，广泛的研究已证明螯合银是一种有效的抗菌剂。抗微生物作用据报道是通过阻断微生物的呼吸发生。螯合银不同于胶态银。螯合意味着是在溶液中，而胶态意味着是在混悬液中。在胶态混悬液中，微小的银粒子是悬浮或漂浮在水性溶液中，不是均匀地分散在整个的液体中。当一种金属离子与双齿的配体形成杂环时，螯合溶液形成。双齿配体的实例是碳酸盐和草酸盐离子以及乙二胺。作为一般的规则，五或六元环是有利的。因为螯合银离子被化学结合到非金属离子，它们是均匀分散在整个溶液的。螯合银离子均匀分散在整个溶液中极大增强它们在整个过程中的抗菌作用和效力。
授予James W.Van Leuven的三个美国专利包括一种液体抗菌剂，其银离子与聚果胶酸钠螯合。Van Leuven的美国专利No.4,184,974公开了一种液体清洁剂、润滑剂和局部杀生物的物质，含有月桂基二乙醇酰胺、丙二醇、甘油、聚果胶酸钠、一种水溶的去污剂、银离子、和足量碱以维持pH在7.2-7.8的范围、和蒸馏水。美国专利No.4,267,168是公布的’974专利的继续申请，公开了一种治疗人的外部组织的方法，该方法使用美国专利No.4,184,974公开的液体清洁剂、润滑剂和局部杀生物剂。美国专利No.4,289,758是公布的’168专利的继续申请，公开了一种液体杀生物的物质，含有大约100-400ppm聚果胶酸钠、大约13-250ppm银离子、大约4-8％甘油、足量的碱以维持pH7.2-7.8的范围、和蒸馏水。
对于Van Leuven专利的溶液，一个已知的缺点是，螯合银离子缺乏长期的稳定性。这种稳定性显然是由于聚果胶酸酯分子的逐渐分解或是逐渐形成额外的化合物所致，所述化合物其对银离子比聚果胶酸酯分子有更大的亲和力。这种分解在极度冷或热和光的存在下加速。越热或摄入的光的能量越强，分解越快。
                       发明概述
一种液体抗菌剂和清洁剂，具有改良的长期稳定性，包括至少以下主要成分：去离子水，银离子，聚果胶酸酯，和乙二胺四乙酸(EDTA)。溶液的长期稳定性增加和抗菌作用延长是由强化银离子的螯合键提供的。目前优选的清洁剂实施例也包括甘油、1，2-丙二醇(也就是丙二醇)，和至少一种选自以下任一族的表面活性剂，其选自烷基硫酸盐、磺酸盐、烷醇酰胺、甜菜碱、氧化胺、肌氨酸盐和磺基琥珀酸盐。本技术优选的实施例事实上已经制备和销售，包括以下重量比的成分：大约70-80％去离子水，2.5-5.5％TEA十二烷基苯磺酸盐；1.5-3.25％月桂酰胺DEA；5.5-11.5％甘油；3.0-5.5％丙二醇；0.005-0.015％聚果胶酸钠和0.005-0.015％EDTA；0.01-0.03％银离子；和足以达到pH值在7.2-7.8的范围的一种缓冲化合物。
                       发明详述
一种液体抗菌剂和清洁剂，具有改良的长期稳定性，包括至少以下主要成分：去离子水，银离子，聚果胶酸酯，和乙二胺四乙酸(EDTA)。EDTA添加到制剂中提高了螯合键的强度，由此增加它的长期稳定性。
目前本技术优选的实施例也包括甘油、1，2-丙二醇(也就是丙二醇)，和至少一种选自以下任一族的表面活性剂，其选自烷基硫酸盐、磺酸盐、烷醇酰胺、甜菜碱、氧化胺、肌氨酸盐和磺基琥珀酸盐。本技术优选的实施例事实上已经制备和销售，包括以下重量比的成分：大约70-80％的去离子水，2.5-5.5％TEA十二烷基苯磺酸盐；1.5-3.25％月桂酰胺DEA；5.5-11.5％甘油；3.0-5.5％丙二醇；0.005-0.015％聚果胶酸钠和0.005-0.015％EDTA；0.01-0.03％银离子；和足以使pH值达到7.2-7.8范围内的缓冲化合物。当pH小于7.0，聚果胶酸酯会胶凝。一定数量的强碱-弱酸类型的缓冲化合物可以用于该组合物，只要它们可以与其它存在的原料相容。因为氢氧化铵十分适合与聚果胶酸钠、EDTA、和硝酸银的组合物使用，它可以作为缓冲化合物使用并占组合物总重的大约0.03％。
聚果胶酸钠可以商购，或可以通过用碳酸钠处理果胶以便溶解它来制备。聚果胶酸酯迅速地与碱土金属离子如钙和镁螯合。
为了提供水性环境，其有益于银离子与EDTA或聚果胶酸酯螯合复合物的形成，首先配制一种含有硝酸银和氨水(以氢氧化铵的形式)的溶液，然后将EDTA和/或聚果胶酸酯加入到该溶液中。许多复合离子和中性分子已知共存于有硝酸银和氨水混合物在内的平衡状态。由于EDTA和聚果胶酸酯比铵和氢氧离子对银离子更具有亲和力，通过预先将氨加到硝酸银中，可促进与前两种化合物形成螯合复合物。以下的化学计量是螯合过程中一般的应用配方。A和B部分分别制备，然后混合到一起形成螯合银溶液。
A部分
硝酸银                            30-60克
去离子水                          2升
氢氧化铵(何种浓度？)              200毫升
B部分
乙二胺四乙酸(EDTA)
聚果胶酸钠                      19.5克
甘油                            200毫升
去离子水                        5升
由犹他州的Provo的LaSal实验室制作原料安全数据表指出根据本发明制备的螯合银制剂，符合OSHA危害接触标准，29 CFR1910.1200，不含有危险成分。同样确定，从摄食的观点看，制剂几乎没有毒性，大鼠急性口服LD50水平大约为5克每公斤体重。同时也确定该制剂没有原发的皮肤刺激，和只有对小动物眼睛轻微到中等的刺激。
虽然本发明的银制剂可能通常对脊椎动物无害，但它们具有显著的抗菌性质。为了研究本发明螯合银制剂的抗菌有效性，试验已由LaSal实验室进行。一个典型的试验在2003年8月20进行，对一种普通的传染性细菌使用以下的方法确定根据本发明制备螯合银溶液的有效性。
1.测试有机体。
测试混悬液是这样制备的，取5ml培养物，该培养物是在ToddHewitt肉汤中生长的金黄色葡萄球菌ATCC#6538，置于50ml圆锥形聚丙烯离心管中在37℃、和250PRM下震动20小时。该培养物由离心法成团，用5ml无菌18MΩ水洗涤，再次离心，然后再混悬于有1ml无菌水的最后容器中。
2.中和剂。
用以下成分配置一种中和溶液：吐温80--10.0％，卵磷脂--1.7％，胰蛋白胨--1.0％，硫代硫酸钠--0.6％，酵母提取物--0.5％，NaCl--0.5％，半胱氨酸--0.04％，和Tamol--6.0％。该溶液用高压灭菌法灭菌。
3.混悬液测试步骤。
3.1将9.9ml等份的螯合银溶液置于无菌的20mm×150mm的管中。该管平衡于20℃水浴中。
2.2该管用100μl的测试生物混悬液接种，计零。以强力混合该管以得到均匀的混悬液。
3.3经过精确的三分钟后，1ml生物/螯合银溶液被移至9ml中和剂中。两分钟后，中和的混悬液在生理盐水(PPS)中连续稀释至1∶10。
3.4用膜滤法测定在每个稀释管中的活生物体的数量。从选定的稀释管中取1ml等份用平皿培养双份。该膜用大约100mnl的无菌PPS洗涤，然后移至Trypticase Soy Agar培养皿中。该培养皿在37℃下孵育21小时。
3.5计数在每个过滤器上的菌群数量，然后计算对数减少值和死亡值百分比。
4.对照。
4.1待测混悬液的滴度是用膜滤法测定选定的测试生物体混悬液的1∶10PSS稀释液进行的。
4.2中和剂对照这样测定，加1ml消毒剂到9ml中和剂，然后以100μl的1∶1×104稀释的测试混悬液接种该管。该管静置20分钟，然后稀释至1∶10和1∶100。从这三管取1ml等份用膜滤法测定两次。
4.3在测试中使用的每个溶液的无菌对照，由过滤1ml样品完成，一式两分，取100ml等份过滤用于PSS对照。
5.结果
5.1效价
稀释：                       10-7       10-8       10-9
菌落形成单位的数量(CFU)：    TNC         96          6
                             TNC         96          9
(TNC＝总数不计)
5.2混悬液测试
葡萄球菌/螯合银稀释混悬液：    10-1       10-2     10-3
3分钟后每个培养皿的CFU：       108         15        0
                               117         9         0
5.3中和剂对照
3分钟后每个培养皿的CFU：       未稀释      10-1     10-2
                               TNC         TNC       128
                               TNC         TNC       112
5.4无菌对照
双份测定每个溶液产生以下CFU计数：
PSS    水      中和剂       螯合银溶液
0      0       0            0
0      0       0            0
6.结果
滴定结果显示，活葡萄球菌浓度为每毫升原始混悬液中有9.60×109有机体。用该混悬液接种9.9ml螯合银产生每毫升测定管中有9.60×107有机体的初始浓度。
根据这些过程的结果，允许使用以下公式计算对数减少(LR)值：
LR＝-Log(S/So)，其中S＝经过三分钟后的活有机体浓度；和So＝计零时活有机体的初始浓度。
PK＝(1-(S/So))×100。
使用这些公式，计算出LR值为4.93，PK值为99.9988。总之，螯合银溶液在三分钟内产生几乎5-log的金黄色葡萄球菌减少值。这等于或大于目前使用的许多其它局部抗菌剂的抗菌活性。
虽然本发明已经公开了作为具有特别成分的配方，每种描述有特别的浓度范围，本领域技术人员的改变或调整显然没有超出如以下权利要求的本发明的范围和精神。

Claims (5)

1.一种具有增强的长期稳定性的液体抗菌清洁剂，含有：
70-80％重量的水；
至少一种表面活性剂化合物，占4.0-8.75％重量；
5.5-11.5％重量的甘油；
3.0-5.5％重量的1，2-丙二醇；
0.005-0.015％重量的聚果胶酸钠；
0.005-0.015％重量的乙二胺四乙酸；
0.01-0.03％重量的银离子；和
足量的碱以使pH值达到7.2-7.8的范围；
其中在液体抗菌清洁剂中各组分含量之和为100％。
2.权利要求1的抗菌清洁剂，其中所述至少一种表面活性剂选自烷基硫酸盐、磺酸盐、烷醇酰胺、甜菜碱、氧化胺、肌氨酸盐和磺基琥珀酸盐。
3.权利要求1的抗菌清洁剂，其中所述至少一种表面活性剂含有：2.5-5.5％重量的TEA十二烷基苯磺酸盐和1.5-3.25％重量的月桂酰胺DEA。
4.一种具有增强的长期稳定性的液体抗菌清洁剂，含有70-80％重量的水，0.01-0.03％重量的银离子、0.005-0.015％重量的聚果胶酸酯、0.005-0.015％重量的乙二胺四乙酸以及占4.0-8.75％重量的至少一种表面活性剂化合物；5.5-11.5％重量的甘油；3.0-5.5％重量的1，2-丙二醇；和足量的碱以使溶液的pH值达到7.2-7.8的范围；其中在液体抗菌清洁剂中各组分含量之和为100％，并且所述银离子与所述聚果胶酸酯和乙二胺四乙酸两者螯合。
5.一种制备液体抗菌剂的方法，包括以下步骤：
混合水、银离子和氨水以形成银铵复合溶液；
混合乙二胺四乙酸、聚果胶酸钠、甘油和水以形成螯合溶液；
混合银铵复合溶液与螯合溶液以形成螯合银溶液；
向螯合银溶液添加至少一种表面活性剂化合物；
向螯合银溶液添加1，2-丙二醇；和
向螯合银溶液添加足量的碱以使溶液的pH值达到7.2-7.8的范围；
其中所述液体抗菌剂含有：
70-80％重量的水；
0.01-0.03％重量的银离子；
0.005-0.015％重量的聚果胶酸钠；
0.005-0.015％重量的乙二胺四乙酸；
4.0-8.75％重量的至少一种表面活性剂化合物；
5.5-11.5％重量的甘油；
3.0-5.5％重量的1，2-丙二醇；和
0.03％重量的氨水；
其中在液体抗菌剂中各组分含量之和为100％。
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