CN113509433A - 美白组合物及其制剂和美白方法 - Google Patents
美白组合物及其制剂和美白方法 Download PDFInfo
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- CN113509433A CN113509433A CN202111021655.3A CN202111021655A CN113509433A CN 113509433 A CN113509433 A CN 113509433A CN 202111021655 A CN202111021655 A CN 202111021655A CN 113509433 A CN113509433 A CN 113509433A
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Abstract
本发明涉及化妆品技术领域,特别涉及美白组合物及其制剂和美白方法。该美白组合物由透明质酸酶和美白成分组成。本发明采用透明质酸酶与干细胞外泌体悬液或其它美白成分混均匀使用,透明质酸酶可促使细胞通透性增加,干细胞外泌体悬液或其它美白产品使面部肌肤细胞外基质胶原蛋白和弹性蛋白活性恢复,激活抗氧化系统,促进肌肤的代谢更新,达到一个黑色素和自由基的清除、美白细致肌肤的目的。
Description
技术领域
本发明涉及化妆品技术领域,特别涉及美白组合物及其制剂和美白方法。
背景技术
黑色素是一种生物色素,是酪氨酸或3,4-二羟苯丙氨经过一连串化学反应所形成,动物、植物与原生生物都有这种相同色素。黑色素通常以聚合的方式存在。黑色素其实是一种氨基酸衍生物,在每个人的体内都有,其是以一种特殊的细胞即黑色素细胞生成并且储存在其中,正是由于黑色素的存在,皮肤才有了颜色。20-25岁是黑色素沉淀形成的活跃期。它们存在于皮肤基底层的细胞中间,而是一种叫“黑色素原生物质”的物质,也叫做“色素母细胞”。色素母细胞分泌麦拉宁色素,当紫外线(B波、A波)照射到皮肤上(B波即UVB作用于皮肤基底层),肌肤就会处于“自我防护”的状态,即由紫外线刺激麦拉宁色素,激活酪氨酸酶的活性,来保护我们的皮肤细胞。多巴其实就是黑色素的前身,经酪氨酸氧化而成,释放出黑色素。黑色素又经由细胞代谢的层层移动,到了肌肤表皮层形成雀斑、晒斑、黑斑等形状了。
导致皮肤暗沉、多斑的原因很多,主要影响因素为自身内环境和外界环境两大因素所影响。从自身内环境看,由于生活不规律、饮食习惯不好等原因影响自身机体代谢功能的下降,导致身体黑色素堆积不能及时的排除体外,在身体内长期积累而引发的;从外环境看,由于生活所处的紫外线过强,引发皮肤的暗沉、多斑等。
目前市面有很多面部美白产品,包括外用护肤的美白化妆品、注射用的美白制剂、排毒美白养颜的口服产品。主要通过外在防护或改善身体的代谢机能,使面部皮肤达到美白的效果。但对特定部位作用不明显,周期长,效果慢,属于内调。面部美白注射制剂的美白主要原理是通过注射制剂中所含成分,抑制细胞氧化、快速分解人体肌肤的黑色素、黄色素、修复受损细胞,从而改善缺氧性晦暗、粗糙皮肤等现象。现在市场上的美白针主要成分是传明酸(TXA)和天然维生素C(NVc)、谷胱甘肽,传明酸可以控制黑色素的酵素作用,减少黑色素形成;谷胱甘肽有助于身体排毒,也可帮助细胞抗氧化;维他命C美白成份,具抗氧化作用,还原黑色素。传明酸只能改善黑斑不再变浓、扩大及增加,从而防止和改善皮肤的色素沉积,但积累的黑色素改善作用不明显;维生素C具有抗氧化、促进胶原蛋白合成等效果,每天至少需要摄入维生素C 1000mg以上,但维生素C长期大剂量摄入,会促进血液系统铁的吸收,引起高铁红细胞贫血,还可减少肠道对维生素B12的吸收,使巨幼红细胞性贫血加速恶化,如每天用量超过5克可导致溶血,重者可以致命,长期大剂量服用后突然停药,可能造成反跳性坏血病,对消化系统、泌尿系统等造成一定的损伤;添加的天然草本精华成分,因人而异,不一定能适合所有个体使用。以上抗氧化的原理可以短时间抑制黑色素的扩散,但随着黑色素的逐渐产生,还是会恢复到之前的状态,稳定性不好。而且,现有的美白制剂注射途径大多是静脉注射,注射频率较高。
发明内容
有鉴于此,本发明提供了美白组合物及其制剂和美白方法。该美白组合物可改善肌肤对美白成分的吸收效果,美白效果显著且稳定,无需频繁使用。
为了实现上述发明目的,本发明提供以下技术方案:
本发明提供了一种美白组合物,该美白组合物由透明质酸酶和美白成分组成。
作为优选,美白成分选自但不限于干细胞外泌体、传明酸、维生素C、谷胱甘肽、重组人类生长因子中的一种或几种。
透明质酸酶(hyaluronidase,HAase)是能使透明质酸产生低分子化作用酶的总称,为一种能水解透明质酸的酶(透明质酸为组织基质中具有限制水分及其它细胞外物质扩散作用的成分),用于人体能暂时降低细胞间质的粘性,可促使皮下输液、局部积贮的渗出液或血液加快扩散而利于吸收,为一种重要的药物扩散剂。临床用作药物渗透剂,促进药物的吸收,促进手术及创伤后局部水肿或血肿消散。
外泌体是一种能被大多数干细胞分泌的微小囊泡,直径大约在30-150nm,由脂质、蛋白质和核酸组成。外泌体一旦通过胞吐作用从干细胞中释放,复杂的混合因子能作为信号分子传递给其他细胞,从而改变其他细胞的功能。外泌体被包裹在坚硬的双层膜中,双层膜保护外泌体的内容物,使外泌体能够在组织中长距离移动。外泌体因在上皮组织的增殖、迁移、再生、炎症和瘢痕控制等方面的作用,紫外线损伤和老化会影响细胞外基质胶原蛋白和弹性蛋白活性,这两者都是防止皮肤脱水以及保持皮肤紧致和弹性的关键。外泌体可以恢复老年人皮肤成纤维细胞的功能,有相关报道指出经iPSC外泌体预处理的皮肤成纤维细胞能够抵抗UVB的光老化,并且不会过度表达基质降解酶MMP-1/3,相反,它们会像年轻的成纤维细胞一样高表达I型胶原。
传明酸(Tranexamic acid),又名氨甲环酸、凝血酸、止血环酸,分子式为C8H15NO2,化学名为反-4-氨甲基环已烷甲酸,为白色结晶性粉末,无臭,味微。在水中易溶,在乙醇、丙酮、三氯甲烷或乙醚中几乎不溶。其退黑除斑的功效比维生素C高约50倍,是果酸的近10倍左右。
维生素C,又称维他命C,是一种多羟基化合物,化学式为C6H8O6。结构类似葡萄糖,其分子中第2及第3位上两个相邻的烯醇式羟基极易解离而释出H+,故具有酸的性质,又称L-抗坏血酸。维生素C具有很强的还原性,很容易被氧化成脱氢维生素C,但其反应是可逆的,并且抗坏血酸和脱氢抗坏血酸具有同样的生理功能,但脱氢抗坏血酸若继续氧化,生成二酮古乐糖酸,则反应不可逆而完全失去生理效能。维生素C为抗体及胶原形成,组织修补(包括某些氧化还原作用),苯丙氨酸、酪氨酸、叶酸的代谢,铁、碳水化合物的利用,脂肪、蛋白质的合成,维持免疫功能,羟化5-羟色胺,保持血管的完整,促进非血红素铁吸收等所必需,同时维生素C还具备有抗氧化,抗自由基,抑制酪氨酸酶的形成,从而达到美白、淡斑的功效。
谷胱甘肽(glutathione,r-glutamyl cysteingl+glycine,GSH)是一种含γ-酰胺键和巯基的三肽,由谷氨酸、半胱氨酸及甘氨酸组成,存在于几乎身体的每一个细胞。谷胱甘肽能帮助保持正常的免疫系统功能,并具有抗氧化作用、整合解毒作用。半胱氨酸上的巯基为其活性基团,易与某些药物、毒素等结合,使其具有整合解毒作用。谷胱甘肽不仅可用于药物,更可作为功能性食品的基料,在延缓衰老、增强免疫力、抗肿瘤等功能性食品广泛应用。主要功效为抗氧化、整合解毒、美白、淡斑。
人类生长因子:HGF(Human growth factor)即人类生长因子。随着基因科学的发展,科学家们成功地从植物中提取生长因子。它是一种生物活性肽复合物,有多个特殊氨基酸组成肽链,进入人体血液后,促进我们的下丘脑自行分泌人体生长素,而人体生长素对我们的生长发育、各种蛋白质的合成、细胞的修复和分裂繁殖、激发胸腺恢复功能,增强人体免疫力、促进新陈代谢、增强人体排毒功能、再造老器官功能、抗击衰老、整体增进健康水平有着决定性的作用。生长因子作为特殊的氨基酸肽,还有着更多有益身体的功能,在医学上对愈合伤口、重症病人的恢复及荷尔蒙治疗方面有着广泛的运用。因而,人类生长因子被称为人类身体的黄金物质。
本发明提供一种复合注射制剂,由I型和I型制剂组成的复合I/II型个性化面部注射美白制剂套组,I型制剂含有透明质酸酶,可暂时降低细胞间质的粘性,促进药物的吸收,促进注射后局部水肿或血肿消散,同时具有保湿作用,能很好的维持治疗后期的稳定,防止黑色素重新聚集;II型制剂为干细胞外泌体悬液或其它美白成分,可直接促进纤维细胞增殖和胶原蛋白合成,修复皮肤损伤,抑制MMPs的活性,减少皮肤基质降解,同时可以激活抗氧化系统,促进抗氧化物质的增加,达到全面抗氧化,保护和美白皮肤。复合I/II型个性化面部注射美白制剂通过皮下注射,能使美白营养物快速直接到达基底层,促进面部血液循环及新陈代谢,修复面部肌肤暗沉无光泽,安全性高,见效快,效果维持时间长。
本发明还提供了一种美白制剂,该美白制剂包括透明质酸酶溶液和含有美白成分的溶液组成。
作为优选,透明质酸酶溶液的溶剂为生理盐水,透明质酸酶溶液的浓度为1~10U/mL。
作为优选,含有美白成分的溶液中的美白成分选自但不限于干细胞外泌体、传明酸、维生素C、谷胱甘肽、重组人类生长因子中的一种或几种。
作为优选,含有美白成分的溶液为干细胞外泌体悬液,干细胞外泌体悬液中干细胞外泌体的密度为450~500μg/mL。
作为优选,干细胞外泌体悬液的制备方法为:将间充质干细胞采用无血清完全培养基进行传代培养,收集传代培养获取的培养上清,采用30~50μm的细胞过滤器过滤,将过滤液离心,收集离心上清,采用0.2~0.3μm的过滤材料过滤。
作为优选,间充质干细胞选自但不限于脐带间充质干细胞、胎盘间充质干细胞、骨髓间充质干细胞、羊膜间充质干细胞、脂肪间充质干细胞中的一种或几种。
在本发明提供的具体实施例中,采用40μm细胞过滤器过滤。
作为优选,离心的条件为:0~4℃、8000~12000g离心10~30min。
在本发明提供的具体实施例中,离心的条件为:4℃、10000g离心20min。
在本发明提供的具体实施例中,采用0.22μm的过滤材料过滤。
作为优选,干细胞外泌体悬液中干细胞外泌体的密度450~500μg/mL。
本发明还提供了一种美白方法,包括:将美白产品施于面部皮肤;美白产品为上述美白组合物,或者为上述美白制剂。
作为优选,该美白产品的施用方式为注射。
本发明提供了美白组合物及其制剂和美白方法。该美白组合物由透明质酸酶和美白成分组成。本发明具有的技术效果如下:
采用透明质酸酶与干细胞外泌体悬液或其它美白成分混均匀使用,透明质酸酶可促使细胞通透性增加,干细胞外泌体悬液或其它美白产品使面部肌肤细胞外基质胶原蛋白和弹性蛋白活性恢复,激活抗氧化系统,促进肌肤的代谢更新,达到一个黑色素和自由基的清除、美白细致肌肤的目的。
具体实施方式
本发明公开了美白组合物及其制剂和美白方法,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。特别需要指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明。本发明的方法及应用已经通过较佳实施例进行了描述,相关人员明显能在不脱离本发明内容、精神和范围内对本文所述的方法和应用进行改动或适当变更与组合,来实现和应用本发明技术。
本发明中所用试剂或仪器均可由市场购得。
下面结合实施例,进一步阐述本发明:
实施例1复合I/II型个性化面部注射美白制剂套装的制备
本发明将I型制剂与II型制剂组合在一起,形成复合I/II型个性化面部注射美白制剂套装,步骤如下:
1、I型制剂制备:
将60U的透明质酸酶溶于30mL注射级的生理盐水中,配制成2U/mL的透明质酸酶溶液,分装成2.5mL/支。
2、II型制剂制备:
(1)脐带采集
1)供者来源于已取得《医疗机构执业许可证》的具有供者筛查能力的医院。
2)供者选择
对供者的一般信息、既往病史、家族史等进行资料采集,要求供者无家族遗传病、无恶性肿瘤疾病史、无感染性疾病、无自身免疫性疾病、无特定病毒感染(HIV、HBV、HCV、梅毒螺旋体、EBV、巨细胞病毒)身体健康的40岁以下供者,采集前需供者自愿提供近3个月的三甲医院体检报告。
3)供者体检报告的要求
采集标本前,供者需提供近6个月的三甲医院体检报告,若体检报告中具有完整的感染四项结果,除乙肝表面抗体可阳可阴外,其他项目报阳,则拒绝接收此标本。
4)若不能提供近6个月完整的体检报告,则需要提供5mL母血(或脐血)送至质检部进行病毒(HIV、HBV、HCV、梅毒螺旋体)检测,除乙肝表面抗体可阳可阴外,其他项目报阳,则废弃此标本。
5)提供的体检报告必须清晰、准确和完整,模糊的体检报告拒绝接收。
采集前,采集物的采集必须得到供者或其监护人的同意,签署并填写《脐带间充质干细胞采集信息表》
6)采集前,采集相关人员检查脐带采集套装内物品是否齐全,脐带采集溶液是否在有效期内,尤其注意无菌脐带保存液是否无色透明,如有浑浊、渗漏及瓶口松动,应马上更换。
7)采集工作应由医院的医护人员实施。采集人员为持有医师或护士执业证书,并经过相应的培训后方能进行采集。
8)胎儿娩出后,常规断脐,于10分钟内在脐带靠近脐带端2-3cm处,用止血钳夹住脐带,然后在靠近止血钳处用丝线结扎,在结扎处和止血钳之间,用手术剪将脐带剪断,脐带的另一端也用细丝线结扎,两个结扎点之间的脐带长度须不少于15cm;同时脐带颜色白净,脐带内有少量未凝固脐血,脐带无凝集血块,无肿胀,未见颜色异常。
9)清洗脐带,并迅速放入无菌脐带保存液内,密封无菌采集瓶。
(2)脐带干细胞获取
向15cm培养皿内倒入100mL预冷的生理盐水,用平镊取出脐带置于其内,清洗脐带表明血渍,用手术剪剪去包扎脐带首尾的脐带夹连同脐带1cm,用止血钳夹止脐带一端,用平镊挤掉血管内血液。
1)用平镊将脐带转移到新的15cm培养皿,倒入100mL预冷的75%酒精,浸泡约30s。
2)用平镊将济带转移到新的15cm培养皿,倒入100mL生理盐水,洗去酒精,此步骤重复一次。
3)用直头手术剪把脐带剪成2cm长的小段。取两把组织镊,夹取一小段脐带,选取两条动脉之间为切入点剥离脐带外膜,取另一把平镊,穿入静脉血管,剥离静脉,再把两条动脉剥离,弃脐带外膜及血管,注意保留血管外周的组织,把获得的组织撕成块。
4)按3)步骤处理所有的脐带小段,把获得的组织转移至烧杯中,添加少量完全培养基至离心管后,用手术剪剪至50mm3大小。
5)根据脐带的长度选择接种培养瓶数量,每4mL脐带长度接种1个T-175培养瓶,用镊子取相应组织块平铺到培养团中,待组织块固定于培养皿上。
6)在烧杯中加入无血清完全培养基,充分混匀后用移夜管加入25mL至铺有组织块的培养瓶中,轻轻摇晃培养团,使培养基均匀分布于培养瓶中。
7)培养7至14天可以获得脐带间充质干细胞。
8)将获取的间充质干细胞进行无血清完全培养基传代培养。
(3)干细胞外泌体悬液获取
1)收集传代培养获取的培养上清。
2)使用40um细胞过滤器过滤,收集过滤液。
3)过滤液离心管配平后置于离心机内,4℃10000g离心20min,收集离心上清。
4)离心上清0.22um过滤,将过滤液分装成2.5mL/支。干细胞外泌体悬液中干细胞外泌体的密度为450μg/mL。
3、美白制剂套装组合
将步骤1和步骤2中制备得到的2.5mL I型制剂和2.5mL II型制剂组成复合I/II型个性化面部注射美白制剂套装。
在使用时,先将复合I/II型制剂套装中的I型制剂和II型制剂混均匀,再进行注射,I型制剂可促使细胞通透性增加,II型制剂使面部肌肤细胞外基质胶原蛋白和弹性蛋白活性恢复,激活抗氧化系统,促进肌肤的代谢更新。
试验例1复合I/II型个性制剂套装效果测试
选择100例不同年龄、脸上都有不同程度黑色素沉和面部肤色暗黄的试验者进行试验,其中50例面部滚针头注射实验,进行额头和面颊注射实施例1美白制剂套装,一次使用量为4mL,两周一次,连续一个月,有36例脸上褐斑消失,肤色变白,剩下的14例面部肤色均有不同程度的变白,并且没有过敏现象发生。另外50例进行等量生理盐水面部滚针头注射,连续一个月,并没有出现明显变化。
实施例2复合I/II型个性化面部注射美白制剂套装的制备
本发明将I型制剂与II型制剂组合在一起,形成复合I/II型个性化面部注射美白制剂套装,步骤如下:
1、I型制剂制备:
将60U的透明质酸酶溶于30mL注射级的生理盐水中,配制成2U/mL的透明质酸酶溶液,分装成2.5mL/支。
2、II型制剂制备:
将1g传明酸溶于30mL注射级的生理盐水中,配制成0.03g/mL的传明酸溶液,分装成2.5mL/支。
3、美白制剂套装组合
将步骤1和步骤2中制备得到的2.5mL I型制剂和2.5mL II型制剂组成复合I/II型个性化面部注射美白制剂套装。
实施例3复合I/II型个性化面部注射美白制剂套装的制备
本发明将I型制剂与II型制剂组合在一起,形成复合I/II型个性化面部注射美白制剂套装,步骤如下:
1、I型制剂制备:
将60U的透明质酸酶溶于30mL注射级的生理盐水中,配制成2U/mL的透明质酸酶溶液,分装成2.5mL/支。
2、II型制剂制备:
将22.5g维生素C溶于30mL注射级的生理盐水中,配制成0.75g/mL的维生素C溶液,分装成2.5mL/支。
3、美白制剂套装组合
将步骤1和步骤2中制备得到的2.5mL I型制剂和2.5mL II型制剂组成复合I/II型个性化面部注射美白制剂套装。
实施例4复合I/II型个性化面部注射美白制剂套装的制备
本发明将I型制剂与II型制剂组合在一起,形成复合I/II型个性化面部注射美白制剂套装,步骤如下:
1、I型制剂制备:
将60U的透明质酸酶溶于30mL注射级的生理盐水中,配制成2U/mL的透明质酸酶溶液,分装成2.5mL/支。
2、II型制剂制备:
将1.2g谷胱甘肽溶于30mL注射级的生理盐水中,配制成0.04g/mL的谷胱甘肽溶液,分装成2.5mL/支。
3、美白制剂套装组合
将步骤1和步骤2中制备得到的2.5mL I型制剂和2.5mL II型制剂组成复合I/II型个性化面部注射美白制剂套装。
试验例2复合I/II型个性制剂套装效果测试
选择150例不同年龄、脸上都有不同程度黑色素沉和面部肤色暗黄的试验者进行试验,分50例一组,共3组,面部滚针头注射实验,进行额头和面颊注射实施例2-4美白制剂套装,一次使用量为4mL,两周一次,连续一个月。
实施例2,有18例脸上褐斑退淡,剩下的32例面部肤色无明显变白,并且没有过敏现象发生。
实施例3,均出现疼痛感,出现红肿现象。
实施例4,面部肤色均有不同程度的变白,但部分有过敏现象发生。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
Claims (10)
1.一种美白组合物,其特征在于,由透明质酸酶和美白成分组成。
2.根据权利要求1所述的美白组合物,其特征在于,所述美白成分选自干细胞外泌体、传明酸、维生素C、谷胱甘肽、重组人类生长因子中的一种或几种。
3.一种美白制剂,其特征在于,包括透明质酸酶溶液和含有美白成分的溶液组成。
4.根据权利要求3所述的美白制剂,其特征在于,所述透明质酸酶溶液的溶剂为生理盐水,所述透明质酸酶溶液的浓度为1~10U/mL。
5.根据权利要求3所述的美白制剂,其特征在于,所述含有美白成分的溶液中的美白成分选自干细胞外泌体、传明酸、维生素C、谷胱甘肽、重组人类生长因子中的一种或几种。
6.根据权利要求5所述的美白制剂,其特征在于,所述含有美白成分的溶液为干细胞外泌体悬液,所述干细胞外泌体悬液中干细胞外泌体的密度为450~500μg/mL。
7.根据权利要求6所述的美白制剂,其特征在于,所述干细胞外泌体悬液的制备方法为:将间充质干细胞采用无血清完全培养基进行传代培养,收集传代培养获取的培养上清,采用30~50μm的细胞过滤器过滤,将过滤液离心,收集离心上清,采用0.2~0.3μm的过滤材料过滤。
8.根据权利要求7所述的美白制剂,其特征在于,所述间充质干细胞选自脐带间充质干细胞、胎盘间充质干细胞、骨髓间充质干细胞、羊膜间充质干细胞、脂肪间充质干细胞的一种或几种。
9.一种美白方法,其特征在于,包括:将美白产品施于面部皮肤;所述美白产品为权利要求1或2所述的美白组合物,或者为权利要求3至8中任一项所述的美白制剂。
10.根据权利要求9所述的美白方法,其特征在于,所述美白产品的施用方式为注射。
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CN116898791A (zh) * | 2023-09-14 | 2023-10-20 | 山东博森医学工程技术有限公司 | 一种利用干细胞外泌体进行皮肤美白的医美试剂 |
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