CN116509939A - 一种用于抑制皮肤糖化的组合物 - Google Patents
一种用于抑制皮肤糖化的组合物 Download PDFInfo
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- CN116509939A CN116509939A CN202310330604.1A CN202310330604A CN116509939A CN 116509939 A CN116509939 A CN 116509939A CN 202310330604 A CN202310330604 A CN 202310330604A CN 116509939 A CN116509939 A CN 116509939A
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Abstract
本发明公开了一种用于抑制皮肤糖化的组合物，其特征在于，包含五味子提取物，具有抑制胶原蛋白糖化、抑制细胞骨架糖化、抑制细胞内AGEs(晚期糖化产物)生成和抑制AGEs引起的炎症等其中一种或多种功效，可以提供具有改善皮肤状态乃至改善皮肤老化效果的组合物。
Description
技术领域
本发明涉及组合物领域，具体涉及一种用于抑制皮肤糖化的组合物。
背景技术
糖化(glycation)是糖与蛋白质和脂质的非酶反应，由还原糖的羰基与蛋白质的游离氨基反应形成Schiff base、amadori product和mailard product，这些化合物通过一系列反应生成棕色的不可逆晚期糖化产物(AGEs,Advanced Glycation End products)；
因为人体在摄入糖分后代谢糖分时会感到负担，这时损伤细胞的活性氧会增加，从而诱发衰老，这个过程称为糖化，是代谢功能下降的原因之一；
具体地说，糖化是发生在表皮、真皮和血管等多种皮肤组织中的现象，在细胞单位阻碍细胞生长，导致细胞死亡(Apoptosis)和细胞休止(Senescence)阶段，在角质形成细胞中，报道通过NFκB的核内传入提高炎症介导因子的表达，活性氧(ROS)的生成也随之提高，在成纤维细胞中也能提高炎症介导因子及活性氧的产生，降低细胞外基质(ECM、Extracellular matrix)物质胶原蛋白及弹性蛋白的表达，提高细胞外基质水解酶(MMP.Matrix metalloproteinase)的表达，这导致表皮、真皮的细胞再生及持久性降低，并诱发胶原蛋白及弹性蛋白等聚纤维的交联(Cross-linking)，使其对细胞外基质水解酶产生抗性，最终导致皮肤僵硬，弹性降低，细胞外基质无法进行改造，从而诱导皮肤老化；
与一般角质形成细胞(28天周期细胞死亡)不同，如果对Long-live cell(Fibroblast、Brain cells、Retina cells等)和Long-lasting protein(Collagen、Crystallins、Lens、Cornea等)及DNA进行糖化，就会给皮肤带来严重的问题，皮肤内胶原蛋白的糖化，即Long-lasting protein，通过介导相邻胶原纤维和纤维之间的交联(Cross-linking)，降低皮肤弹性，引起皱纹，据悉，胶原蛋白糖化从20多岁开始，每年以3.7％的速度增加，紫外线及活性氧等外部环境也会增加皮肤糖化速度；
用来抑制皮肤老化的抗氧化已经广为人知，目前存在很多化妆品。近年来，晚期糖化产物的积累成为内源性衰老的主要原因而备受关注，在中国美妆市场，以抗糖化而非抗氧化为目标的化妆品也备受关注，在2022In-cosmetics korea的Innovation zone上，也曾展示过一些以皮肤糖化为目标的原料；
但是，对具有适当功效的同时含对人体无害成分的组合物的研究并不尽如人意。
发明内容
本发明要解决的通过常规醇提工艺即可获得一种组合物，并可通过多种抗糖化方法验证该组合物的抗糖化功效，本发明提供一种用于抑制皮肤糖化的组合物，具有抑制胶原蛋白糖化、抑制细胞骨架糖化、抑制细胞内AGEs(晚期糖化产物)生成和抑制AGEs引起的炎症等其中一种或多种功效，可以提供具有改善皮肤状态乃至改善皮肤老化效果的组合物，用以解决现有技术导致的缺陷。
为解决上述技术问题本发明提供以下的技术方案：
一种用于抑制皮肤糖化的组合物，其中，包含五味子提取物。
上述的一种用于抑制皮肤糖化的组合物，其中，还包含黄檗提取物、白薇提取物、三七根提取物中的一种或多种。
上述的一种用于抑制皮肤糖化的组合物，其中，为药物组合物、化妆品组合物、食品组合物中的任一种。
上述的一种用于抑制皮肤糖化的组合物，其中，为片剂、胶囊剂、注射剂、面霜、凝胶、面膜、贴片、喷雾剂、软膏剂、硬膏剂、乳液剂、擦剂、糊剂和卡介苗剂中的任一种。
上述的一种用于抑制皮肤糖化的组合物，其中，为气溶胶、液体、溶液、凝胶、面片、面膜、带、贴片、面霜、药膏、泡沫中的任一种。
上述的一种用于抑制皮肤糖化的组合物，其中，所述食品组合物为片剂、胶囊剂、软膏剂、丸剂、颗粒剂、饮料、减肥棒、巧克力、焦糖、饼干中的任一种。
依据上述本发明一种用于抑制皮肤糖化的组合物提供的技术方案具有以下技术效果：
具有抑制胶原蛋白糖化、抑制细胞骨架糖化、抑制细胞内AGEs(晚期糖化产物)生成和抑制AGEs引起的炎症等其中一种或多种功效，可以提供具有改善皮肤状态乃至改善皮肤老化效果的组合物。
附图说明
图1为五味子提取物晚期糖化产物生成的抑制图。
图2为白薇提取物晚期糖化产物生成的抑制图。
图3为三七根提取物晚期糖化产物生成的抑制图。
图4为五味子提取物胶原糖化抑制结果图。
图5为白薇提取物胶原糖化抑制图。
图6为三七根提取物胶原糖化抑制图。
图7为五味子提取物对胶原糖化的抑制图。
图8为白薇提取物对胶原糖化的抑制图。
图9为三七根提取物对胶原糖化抑制图。
图10为五味子提取物对细胞内细胞骨架糖化的抑制图。
图11为白薇提取物对细胞内细胞骨架糖化抑制图。
图12为三七根提取物对细胞内细胞骨架糖化抑制图。
图13为五味子提取物对细胞内晚期糖化产物表达抑制图。
具体实施方式
为了使发明实现的技术手段、创造特征、达成目的和功效易于明白了解，下结合具体图示，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例是本发明的一部分实施例，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的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。
须知，本说明书所附图式所绘示的结构、比例、大小等，均仅用以配合说明书所揭示的内容，以供熟悉此技术的人士了解与阅读，并非用以限定本发明可实施的限定条件，故不具技术上的实质意义，任何结构的修饰、比例关系的改变或大小的调整，在不影响本发明所能产生的功效及所能达成的目的下，均应仍落在本发明所揭示的技术内容得能涵盖的范围内。
同时，本说明书中所引用的如“上”、“下”、“左”、“右”、“中间”及“一”等的用语，亦仅为便于叙述的明了，而非用以限定本发明可实施的范围，其相对关系的改变或调整，在无实质变更技术内容下，当亦视为本发明可实施的范畴。
一种用于抑制皮肤糖化的组合物，其中，包含五味子提取物。
五味子是五味子树的果实，直径约1cm，深红色，可感受到甜味、酸味、苦味、咸味、辣味，因此被称为五味子，含有甜菊酯、苹果酸、柠檬醛等成分，能增强心脏、降低血压、提高免疫力，被用作强壮剂，有增强肺功能、镇咳祛痰作用，有助于治疗咳嗽、口渴等。
上述用于抑制皮肤糖化的组合物宜进一步包含黄柏提取物、白薇提取物和三七根提取物中的一种或多种。
黄柏，又称黄檗、黄木，黄柏是芸香科(Rutaceae)黄檗属(Genus Phellodendron)中的黄柏(Phellodendron amurense Ruprecht)或黄柏的干燥树皮，《中国药典》将黄皮树称为“川黄柏”，只有黄皮树被认定为黄柏起源植物，树皮作药材，大黄柏(Phellodendronamurense var.latifoliolatum NAKAI)、岛生黄檗(P.insulare NAKAI)、毛黄柏(P.molleNAKAI)的外壳也一起使用，黄柏提取物具有健胃、正藏、收敛、止泻、退热、解毒、镇痛等功效，适用于包括消化不良、腹泻、腹痛、黄疸、肝炎、肝硬化、小便不畅、子宫出血等。
白薇是龙胆木(Gentianales)薄竹李科(Asclepiadaceae)属多年生草本植物，因根系洁白细长而得名白薇。
白薇在秋季或早春时挖根，晒干后用水煎煮药用，将白薇的根晒干后，汉方称为白薇，据悉，在高烧或手脚浮肿时有疗效，根有解热作用，可用于肺结核有潮热的地方或血热、虚热，作为药材，还有消除肺结核引起的吐血、肾炎、尿路感染症和隐性病晚期灼热感的功效，对全身水肿和淋病也有疗效。据悉，分布在韩国、日本、中国、蒙古等地。
三七根为五加科人参属(Panax notoginseng(Burkill)F.H.Chen ex C.H.)的根，有止血作用，可祛除瘀血，用于各种出血症状，即吐血、流鼻血、咳血、大小便出血等，也可用于瘀伤骨折、瘀血引起的腹痛、肿胀、止痛、产后失血过多等症状。药理作用有缩短凝血时间、抗疲劳效果、降低血压、舒缓心跳、增加冠状动脉血流量、溶解脂质作用、增进肝功能、激活免疫功能等报道。
上述的一种用于抑制皮肤糖化的组合物能抑制晚期糖化产物的生成、抑制胶原蛋白交联为宜、抑制细胞内细胞骨架糖化、抑制细胞内晚期糖化产物的表达。
上述的一种用于抑制皮肤糖化的组合物为药物组合物、化妆品组合物、食品组合物中的任何一种为宜。
本发明中的药学组合物可以但不限于从群中选择的剂型，包含片剂、胶囊剂、注射剂、面霜、凝胶、贴剂、喷雾剂、软膏剂、硬膏剂、乳液剂、擦剂、糊剂和卡介苗剂等。
本发明的药物组合物还可包含通常用于制剂的药剂学允许的载体、润滑剂、润湿剂、甜味剂、香味剂、乳化剂、悬浮剂、保鲜剂等。
本发明的药物组合物可口服或非口服，非口服者可采用但不限于静脉内注、皮下注、肌肉注、腹腔注、经皮注、粘膜注和滴眼注、局部注等方式。
本发明药物组合物的适宜剂量可根据制剂化方法、给药方式、患者年龄、体重、性别、病态、摄入食物、给药时间、给药途径、排泄速率和反应感应性等因素进行多种处方，优选地，本发明药物组合物的剂量可以但不限于成人0.0001-100mg/kg(体重)
可添加到本发明的合适成分的选择，将在显然的范围内，在一定程度上依赖于本发明的一种用于抑制皮肤糖化的组合物可被剂型化为溶液、乳液、乳霜、软膏、凝胶、泡沫或经皮贴片。
例如，可以包含着色剂(色素)、着香剂(香料)、悬浮剂、乳化剂、助溶剂、稳定剂、澄清剂、pH调节剂、粘度调节剂、溶剂等，化妆品组合物通常附加的成分或含量等，对同行业人员来说是不言而喻的。
根据本发明的化妆品组合物可以且不限于气溶胶、液体、溶液、凝胶、面片、面霜、药膏、泡沫和粉末中的任一剂型。
例如，乳液与溶液类似，往往比溶液更浓，本质上更舒缓，乳液中一般是水和油混合的，而且酒精往往比溶液少。
例如，乳液含有几乎相同比例的油和水精华，能很好地渗透到皮肤角质层外层，面霜比乳液厚，从容器中取出时保持形状。
此外，本发明组合物可包括在面膜、带或贴片中，但不限于。
上述面膜可以是局部型贴片、片状、纱布状、霜状及凝胶状中的任何一种形式。
上述面膜可以是面膜、手部面膜、足部面膜、小腿面膜、发膜、大腿面膜、腹部面膜、肋部面膜、前臂面膜、下巴面膜及颈部面膜中的任一种选择。
另一方面，适用于食品组合物时，可以以本发明的组合物为有效成分，选定糖类中通常使用的成分，将其剂型化为片剂、胶囊剂、丸剂、颗粒剂、饮料(口服剂)、减肥棒、巧克力、焦糖剂型或饼干类剂型等，也可以附加适当地使用适合于保健食品的功能性原料。
以下通过包括根据本发明的五味子提取物的组合物的具体实施例及实验例来更具体地说明本发明。
实施例1：
五味子提取物制备：
在干燥的五味子50克中加入70％乙醇1升，室温下搅拌3小时，萃取而成。利用滤纸对提取溶液进行过滤回收后，利用减压浓缩器60℃进行减压浓缩，完全去除溶剂，得到粉末状的五味子70％乙醇提取物。
实施例2-3：
白薇及三七根提取物制备：
将干燥后的白薇及三七根粉碎，按照上述实施例1的相同方法分别提取了各自的提取物。
实验例1：
晚期糖化产物生成抑制评价：
在0.1M磷酸缓冲液(Phosphate buffer)(PBS,pH7.4,contained 0.02％sodiumazide)中加入0.5M葡萄糖(glucose)和10mg/mL Bovine Serum Albumin(BSA)，在37℃下进行了反应，加入实施例1-3，诱导非酶性反应21天后，在355-460nm波长测定荧光强度，确定对晚期糖化产物(AGEs)生成的抑制程度，阳性对照组采用氨基胍(Aminoguanidine)；实验结果证实，实施例1抑制了BSA的糖化，如图1-3所示。
实验例2：
胶原糖化抑制评价：
人源成纤维细胞在平板上培养24小时后，用400μM的乙二醛(Glyoxal)和实施例1-3进行5天的培养，用胰蛋白酶(Trypsin)回收细胞，用FBS(Fetal bovine serum)重新悬浊，放入胶原混合物中混合，在Anti-LinkTM(Allvivo Vascular公司)处理的24-孔板上培养1小时后，将含有10％FBS的培养基放入上层，培养24小时后，确定胶原蛋白交联(Cross-linking)的数量。
实验结果证实，实施例1-3抑制了胶原蛋白的结合，如图4-9所示。
实验例3：
细胞内细胞骨架糖化抑制评价：
将人源成纤维细胞在平板上培养24小时后，用400μM乙二醛及实施例1-3处理，培养5天，方法用胰蛋白酶回收细胞，在8mm套膜上培养24小时，取出培养基，用3.7％甲醛(formaldehyde)固定细胞，用0.1％ Triton X-100处理，抗波形纤维蛋白抗体，粘附二次抗体，用激光扫描共焦显微镜Confocal laser scanning microscopy确定波形纤维蛋白(Cytoskeletal fiber)的分布。
实验结果表明，糖化诱导的细胞波形蛋白纤维聚集在核周围，实施例1-3证实了通过波形纤维蛋白的进一步伸展抑制糖化，如图10-12所示。
实验例4：
细胞内晚期糖化产物表达抑制评价：
将人源成纤维细胞在平板上培养24小时后，用400μM的乙二醛和实施例1-3处理，培养5天，去除培养液，用RIPA裂解液(RIPA lysis buffer)溶解细胞后，离心分离获得蛋白，通过BCA蛋白质定量获得了同量的蛋白质样品，然后通过SDS-PAGE对蛋白质进行了不同大小的分离，用电转移(Electrotransfer)将细胞蛋白转移到PVDF膜中，并通过免疫印迹(Immunoblot)确定了晚期糖化产物受体(RAGE，Receptor for advanced glycation endproducts)的表达水平。
实验结果证实，实施例1抑制了晚期糖化产物的表达，如图13所示。
综上，本发明的一种用于抑制皮肤糖化的组合物，具有抑制胶原蛋白糖化、抑制细胞骨架糖化、抑制细胞内AGEs(晚期糖化产物)生成和抑制AGEs引起的炎症等其中一种或多种功效，可以提供具有改善皮肤状态乃至改善皮肤老化效果的组合物。
以上对发明的具体实施例进行了描述。需要理解的是，发明并不局限于上述特定实施方式，其中未尽详细描述的设备和结构应该理解为用本领域中的普通方式予以实施；本领域技术人员可以在权利要求的范围内做出各种变形或修改做出若干简单推演、变形或替换，这并不影响发明的实质内容。
Claims (6)
1.一种用于抑制皮肤糖化的组合物，其特征在于，包含五味子提取物。
2.如权利要求1所述的一种用于抑制皮肤糖化的组合物，其特征在于，还包含黄檗提取物、白薇提取物、三七根提取物中的一种或多种。
3.如权利要求1所述的一种用于抑制皮肤糖化的组合物，其特征在于，为药物组合物、化妆品组合物、食品组合物中的任一种。
4.如权利要求3所述的一种用于抑制皮肤糖化的组合物，其特征在于，为片剂、胶囊剂、注射剂、面霜、凝胶、面膜、贴片、喷雾剂、软膏剂、硬膏剂、乳液剂、擦剂、糊剂和卡介苗剂中的任一种。
5.如权利要求4所述的一种用于抑制皮肤糖化的组合物，其特征在于，为气溶胶、液体、溶液、凝胶、面片、面膜、带、贴片、面霜、药膏、泡沫中的任一种。
6.如权利要求5所述的一种用于抑制皮肤糖化的组合物，其特征在于，所述食品组合物为片剂、胶囊剂、丸剂、颗粒剂、饮料、减肥棒、巧克力、焦糖、饼干中的任一种。
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