CN114533836A - 一种不影响bFGF活性的制剂及其制备方法和应用 - Google Patents
一种不影响bFGF活性的制剂及其制备方法和应用 Download PDFInfo
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- CN114533836A CN114533836A CN202111640427.4A CN202111640427A CN114533836A CN 114533836 A CN114533836 A CN 114533836A CN 202111640427 A CN202111640427 A CN 202111640427A CN 114533836 A CN114533836 A CN 114533836A
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Abstract
本发明涉及医药技术领域,具体涉及一种不影响bFGF活性的制剂及其制备方法和应用。所述不影响bFGF活性的制剂的原料按重量份计,包括马齿苋9~13份、蒲公英8~12份、金银花8~12份、黄柏8~12份、鱼腥草5~9份、黄连6~10份、野菊花5~9份、艾叶8~12份、柴胡6~10份、薄荷叶4~8份、如意草6~10份和冰片4~8份。本发明提供了一种不影响bFGF活性的制剂,制剂原料的各组分协同作用,对伤口创面具有消炎杀菌的作用。而现有技术中的杀菌剂或消毒产品会影响bFGF的生物活性。本发明将制剂与bFGF联用,用于伤口创面的治疗,既可以消炎抗菌,又可以促进创面愈合,具有减少疤痕生成的功效。
Description
技术领域
本发明涉及医药技术领域,具体涉及一种不影响bFGF活性的制剂及其制备方法和应用。
背景技术
创面是正常皮肤(组织)在外界致伤因子如外科手术、外力、热、电流、化学物质、低温以及机体内在因素如局部血液供应障碍等作用下所导致的损害。常伴有皮肤完整性的破坏以及一定量正常组织的丢失,同时,皮肤的正常功能受损。创面修复过程首先需保护创面、防止细菌感染,然后抑制创面出血,最后促进创面细胞层面愈合。然而现有的创面修复产品一般只具有灭菌止血的功效,难以使创面快速愈合。
碱性成纤维细胞生长因子(bFGF)具有促进细胞分裂和增殖、促进血管生成、营养神经元、调节细胞代谢等功能,已广泛应用于五官科、烧伤科、整形美容科等科室,对伤口的修复作用已经通过临床验证。碱性成纤维细胞生长因子(bFGF)是一种促细胞生长作用很强的多肽细胞生长因子,由146个氨基酸组成,分子量为16.5KD,对酸、热敏感,对胰蛋白酶、糜蛋白酶和V8蛋白酶敏感,pH<4时失活,在-70℃可以保存几年,4℃可以保存一周。因此,生长因子具有较强的促进伤口愈合功能,可用于创面修复。
然而,由于创面修复过程需要对伤口进行消毒,在临床常用的消毒产品如酒精、碘酒、双氧水等消毒后的创面环境会使生长因子失活,进而导致其功效下降。当生长因子与阳离子表面活性剂类广谱杀菌剂(如苯扎氯铵)联用时,更会严重影响生长因子的活性。
发明内容
有鉴于此,有必要提供一种不影响bFGF活性的制剂及其制备方法和应用。所述制剂在不影响bFGF活性的基础上,还具有消炎抗菌功能,与bFGF联用既可以消炎抗菌,又可以促进创面愈合,具有减少疤痕生成的功效。
本发明的第一个目的在于提供一种不影响bFGF活性的制剂,所述制剂具有消炎抗菌功能;
本发明的第二个目的在于提供一种不影响bFGF活性的制剂的制备方法;
本发明的第三个目的在于提供一种不影响bFGF活性的制剂的应用,将该制剂与bFGF联用后制备成混合溶液,所述混合溶液可用于创面修复。
为实现上述第一个目的,本发明采取以下的技术方案:
一种不影响bFGF活性的制剂,所述制剂的原料包括马齿苋、蒲公英、金银花、黄柏、鱼腥草、黄连、野菊花、艾叶、柴胡、薄荷叶、如意草、冰片中的至少一种。
进一步的,所述制剂的原料按重量份计,包括马齿苋9~13份、蒲公英8~12份、金银花8~12份、黄柏8~12份、鱼腥草5~9份、黄连6~10份、野菊花5~9份、艾叶8~12份、柴胡6~10份、薄荷叶4~8份、如意草6~10份和冰片4~8份中的至少一种。
优选的,所述制剂的原料按重量份计,包括马齿苋9~13份、蒲公英8~12份、金银花8~12份、黄柏8~12份、鱼腥草5~9份、黄连6~10份、野菊花5~9份、艾叶8~12份、柴胡6~10份、薄荷叶4~8份、如意草6~10份和冰片4~8份。
为实现上述第二个目的,本发明采取以下的技术方案:
一种不影响bFGF活性的制剂的制备方法,所述制备方法为按重量份称取制剂原料,粉碎,制备成混合物,然后加水煎煮、浓缩后过滤。
进一步的,煎煮时混合物与水的比例为1:10~100(g/mL)。
进一步的,浓缩程度为混合物与水的比例为1:1~5(g/mL)。
进一步的,过滤为先经多层纱布过滤,再经0.45μm的滤膜过滤。
优选的,所述多层纱布为10层。
为实现本发明的第三个目的,本发明采取以下的技术方案:
一种不影响bFGF活性的制剂的应用,将制剂与bFGF混合制备成混合溶液。混合溶液可用于创面修复。
进一步的,所述混合溶液进一步制成包括但不限于创可贴、敷料、冻干粉或喷雾剂的产品形式。
进一步的,所述bFGF为人碱性成纤维细胞生长因子或牛碱性成纤维细胞生长因子。
进一步的,制剂与bFGF混合体积比例为(0.5~2):1。
优选的,所述bFGF的浓度为1mg/mL。
进一步的,制剂与bFGF混合后的混合溶液需置于棕色西林瓶中,在2~8℃下储存。
优选的,所述储存时间为0~7d。
进一步的,所述创可贴的制备方法为使用医用平布胶布和吸水垫制成创可贴基础材料,取制剂与bFGF的混合溶液200μL至吸水垫中。
进一步的,所述冻干粉的制备方法为将制剂与bFGF的混合溶液冻干即得。
此外,本发明还提供一种不影响bFGF活性的制剂,所述制剂在与bFGF混合后制备成混合溶液,混合溶液可有效治疗糖尿病足溃疡。
一种不影响bFGF活性的制剂,制剂的原料包括马齿苋、蒲公英、金银花、黄柏、鱼腥草、黄连、野菊花、艾叶、柴胡、薄荷叶、如意草,冰片,栀子,黄芪,姜黄,苍术、路路通、血竭、西洋参、当归、没药、乳香、沉香、西红花中的至少一种。
进一步的,制剂的原料按重量份计,包括1~100份马齿苋、1~100份蒲公英、1~100份金银花、1~100份黄柏、1~100份鱼腥草、1~100份黄连、1~100份野菊花、1~100份艾叶、1~100份柴胡、1~100份薄荷叶、1~100份如意草,1~100份冰片、1~100份栀子、1~100份黄芪、1~100份姜黄、1~100份苍术、1~100份路路通、1~100份血竭、1~100份西洋参、1~100份当归、1~100份没药、1~100份乳香、1~100份沉香、1~100份西红花中的至少一种。
本发明的有益效果为:
本发明提供了一种不影响bFGF活性的制剂,制剂原料的各组分协同作用,对伤口创面具有消炎杀菌的作用。而现有技术中的杀菌剂或消毒产品会影响bFGF的生物活性。本发明将制剂与bFGF联用制备成混合溶液,用于伤口创面的治疗,既可以消炎抗菌,又可以促进创面愈合,具有减少疤痕生成的功效。
附图说明
图1为本发明抑菌环测试图;
图2为本发明生物活性测试图;
图3为本发明实施例2制备的制剂与人碱性成纤维细胞生长因子的混合溶液在不同天数下储存后的抑菌性能图;
图4为本发明实施例2制备的制剂与人碱性成纤维细胞生长因子的混合溶液在不同天数下储存后的生物活性数据图;
图5为小鼠普通创面伤口的在不同治疗天数下的愈合情况。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚,下面将结合本发明实施例,对本发明的技术方案作进一步清楚、完整地描述。需要说明的是,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。
本发明所用人碱性成纤维细胞生长因子为盖扶,规格:20000IU。
实施例1
一种不影响bFGF活性的制剂,所述制剂的原料按重量份计,包括马齿苋10份、蒲公英10份、金银花10份、黄柏10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、柴胡8份、薄荷叶6份、如意草8份和冰片6份。
所述制剂采用如下方法制备:
按上述重量份计,将各个原料粉碎,制备成混合物;然后加水煎煮,混合物与水的比例为1:20(g/mL),浓缩至混合物与水的比例为1:2(g/mL)后过滤;过滤为先经10层纱布过滤,再经0.45μm的滤膜过滤。
实施例2
一种不影响bFGF活性的制剂的应用,将不影响bFGF活性的制剂与bFGF混合,制备成混合溶液。具体为将实施例1制备的不影响bFGF活性的制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到用于创面修复的混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。
实施例3
一种不影响bFGF活性的制剂的应用,将不影响bFGF活性的制剂与bFGF混合,制备成混合溶液。具体为将实施例1制备的不影响bFGF活性的制剂与人碱性成纤维细胞生长因子按体积比为1:1混合,得到用于创面修复的混合溶液,人碱性成纤维细胞生长因子浓度为1mg/mL。
实施例4
一种不影响bFGF活性的制剂的应用,将不影响bFGF活性的制剂与bFGF混合,制备成混合溶液。具体为将实施例1制备的不影响bFGF活性的制剂与人碱性成纤维细胞生长因子按体积比为2:1混合,得到用于创面修复的混合溶液,人碱性成纤维细胞生长因子浓度为1mg/mL。
实施例5
一种创可贴,其制备方法为使用医用平布胶布和吸水垫制成创可贴基础材料,取实施例2制备的用于创面修复的混合溶液200μL至吸水垫中。
实施例6
一种冻干粉,其制备方法为将实施例2制备的用于创面修复的混合溶液冻干即得。
对比例1
一种制剂的应用,将制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。所述制剂的原料按重量份计,包括马齿苋10份、金银花10份、黄柏10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、柴胡8份、薄荷叶6份、如意草8份和冰片6份。所述制剂的制备方法同实施例1。
对比例2
一种制剂的应用,将制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。所述制剂的原料按重量份计,包括蒲公英10份、金银花10份、黄柏10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、柴胡8份、薄荷叶6份、如意草8份和冰片6份。制备方法同实施例1。
对比例3
一种制剂的应用,将制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。所述制剂的原料按重量份计,包括马齿苋10份、蒲公英10份、黄柏10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、柴胡8份、薄荷叶6份、如意草8份和冰片6份。制备方法同实施例1。
对比例4
一种制剂的应用,将制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。所述制剂的原料按重量份计,包括马齿苋10份、蒲公英10份、金银花10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、柴胡8份、薄荷叶6份、如意草8份和冰片6份。制备方法同实施例1。
对比例5
一种制剂的应用,将制剂与人碱性成纤维细胞生长因子按体积比为0.5:1混合,得到混合溶液,其中人碱性成纤维细胞生长因子浓度为1mg/mL。所述制剂的原料按重量份计,包括马齿苋10份、蒲公英10份、金银花10份、黄柏10份、鱼腥草7份、黄连8份、野菊花7份、艾叶10份、薄荷叶6份、如意草8份和冰片6份。制备方法同实施例1。
实验数据
一、抑菌环测试
分别取实施例2、对比例1~5制备的制剂与人碱性成纤维细胞生长因子的混合溶液200μL滴至大小相同的吸水垫中,测试抑菌性能。以10%双氧水200μL为阳性对照组,无菌灭菌水200μL为阴性对照组。抑菌测试方法为:用无菌棉拭子蘸取浓度为5×105cfu/mL~5×106cfu/mL金黄色葡萄球菌悬液,在营养琼脂培养基平板表面均匀涂抹3次,盖好平皿,置室温干燥5min。分别将含有不同混合溶液的吸水垫贴放于平板表面,轻压使其贴紧,盖好平皿,置37℃培养箱培养18h观察结果,用游标卡尺测量抑菌环的直径(包括贴片)并记录。抑菌结果如图1和表1所示,图1和表1中a为阳性对照组,b为阴性对照组,c~h分别对应为实施例2、对比例1~5。
表1
由图1和表1可知,本发明实施例2制备的制剂与bFGF混合溶液抑菌环最大,与阳性对照组抑菌效果相当,说明实施例2制备的混合溶液中各个组分协同作用,具有最好的抑菌效果。
二、生物活性测试
将人碱性成纤维细胞生长因子原液、人碱性成纤维细胞生长因子与双氧水按体积比为1:1混合制备的混合溶液、人碱性成纤维细胞生长因子与75%乙醇按体积比为1:1混合制备的混合溶液、实施例2~4制备的制剂与人碱性成纤维细胞生长因子的混合溶液分别作为样品1~6,取以上样品相同酶活单位剂量用Balb/c 3T3细胞测活性。测试结果如图2所示。
由图2可知,与人碱性成纤维细胞生长因子原液相比,本发明实施例2~4提供的制剂与人碱性成纤维细胞生长因子的混合溶液中生长因子的生物活性无明显差距,仍能在一定程度上保证生长因子的生物活性,而其他的抗菌剂如双氧水、乙醇会严重抑制生长因子的生物活性。
三、储存时间测试
将实施例2制备的制剂与人碱性成纤维细胞生长因子的混合溶液分别在2~8度下分别储存3d、7d、10d、20d和30d,测试经长时间储存后本发明制剂与人碱性成纤维细胞生长因子的混合溶液的抗菌性能以及生物活性。其中抗菌性能测试方法按照《消毒技术规范》2002年版2.1.11.3.2的抑菌性能测试方法进行实验,并按照此标准的公式计算样品对金黄色葡萄球菌的抑菌率。生物活性测试方法同上。测试结果分别如图3和图4所示。
由图3和图4可知,随着储存时间的增加,实施例2制备的制剂与人碱性成纤维细胞生长因子的混合溶液的抑菌性能和生物活性逐渐下降。储存时间超过7天后,生物活性大幅度下降。因此本发明的制剂与人碱性成纤维细胞生长因子的混合后,优选在0~7d内使用。
四、对小鼠普通创面愈合的修复
1.取SPF级C57BL/6小鼠40只,雌雄各半(南方医科大学动物实验中心提供),八周龄,体重17~25g,随机分为5组,每组8只,分别对应5组样品进行试验。
2.建造模型方法:腹腔注射5%水合氯醛0.08mL·(10g)-1麻醉,用酒精棉球消毒后背部皮肤,用无菌手术剪刀剪去直径约1.5cm的圆形皮肤。当天分别用棉签将5组样品均匀涂满创面,不包扎。为防止动物之间打斗摩擦引起伤口感染,小鼠均单笼饲养。
3.组1样品为实施例3中不影响bFGF活性的制剂与bFGF混合制备成的混合溶液,组2样品为生理盐水,组3样品为实施例1中不影响bFGF活性的制剂,组4为20000IU规格的盖扶产品,组5样品为碘伏。小鼠用药时间为3天,每天9:00用药。每天观察伤口的愈合情况,记录伤口开始愈合(创面变硬缩紧)和基本愈合的时间(创面缩小至5mm),并在第3天、第6天、第9天、第12天、第15天分别拍照记录伤口复原情况。第16天小鼠处死。实验结果分别如表2和图5所示。
从表2和图5可知,与组2样品相比,组4、组3以及组1的样品均能显著缩短伤口创面开始愈合时间,特别是组1的样品能显著缩短基本愈合时间。
表2
*表示P<0.05,**表示P<0.01。
以上所述实施例仅表达了本发明的几种实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
Claims (10)
1.一种不影响bFGF活性的制剂,其特征在于,制剂的原料按重量份计,包括马齿苋9~13份、蒲公英8~12份、金银花8~12份、黄柏8~12份、鱼腥草5~9份、黄连6~10份、野菊花5~9份、艾叶8~12份、柴胡6~10份、薄荷叶4~8份、如意草6~10份和冰片4~8份。
2.权利要求1所述的不影响bFGF活性的制剂的制备方法,其特征在于,所述制备方法为按重量份称取制剂原料,粉碎,制备成混合物,然后加水煎煮、浓缩后过滤。
3.根据权利要求2所述的不影响bFGF活性的制剂的制备方法,其特征在于,煎煮时混合物与水的比例为1:(10~100);浓缩程度为混合物与水的比例为1:(1~5)。
4.根据权利要求3所述的不影响bFGF活性的制剂的制备方法,其特征在于,过滤为先经多层纱布过滤,再经0.45μm的滤膜过滤。
5.权利要求1所述的不影响bFGF活性的制剂或权利要求2~4任意一项所述的制备方法制备的不影响bFGF活性的制剂的应用,其特征在于,将制剂与bFGF混合制备成混合溶液。
6.根据权利要求5所述的不影响bFGF活性的制剂的应用,其特征在于,所述混合溶液进一步制成包括但不限于创可贴、敷料、冻干粉或喷雾剂的产品形式。
7.根据权利要求5所述的不影响bFGF活性的制剂的应用,其特征在于,制剂与bFGF混合体积比例为(0.5~2):1。
8.根据权利要求5所述的不影响bFGF活性的制剂的应用,其特征在于,所述bFGF的浓度为1mg/mL。
9.根据权利要求5所述的不影响bFGF活性的制剂的应用,其特征在于,制剂与bFGF混合后的混合溶液需置于棕色西林瓶中,在2~8℃下储存。
10.根据权利要求5所述的不影响bFGF活性的制剂的应用,其特征在于,所述创可贴的制备方法为使用医用平布胶布和吸水垫制成创可贴基础材料,取混合溶液200μL至吸水垫中;所述冻干粉的制备方法为将混合溶液冻干即得。
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