CN115279387A - 抑制tslp及治疗或改善皮肤炎症疾病的包括含龙眼肉组合生药提取物的局部用组合物及其用途 - Google Patents
抑制tslp及治疗或改善皮肤炎症疾病的包括含龙眼肉组合生药提取物的局部用组合物及其用途 Download PDFInfo
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Abstract
本发明涉及用于抑制胸腺基质淋巴细胞生成素(TSLP)的表达或者治疗并改善皮肤炎症疾病的包含龙眼肉、藁本及远志的组合生药提取物作为有效成分的局部用药物组合物及化妆品组合物。
Description
技术领域
本发明涉及用于抑制胸腺基质淋巴细胞生成素(TSLP)及治疗或改善皮肤炎症疾病的包含含有龙眼肉(Longanae Arillus)的组合生药提取物的局部用组合物及其用途。
背景技术
特应性皮炎、银屑病等慢性复发性皮肤炎症疾病(特应性皮炎)的病因多种多样,很难治疗。
尤其,一部分特应性皮炎因丝聚蛋白基因发生突变而损伤皮肤屏障,因细菌感染或螨虫源性异物感染引起严重的炎症反应。
除上述基因因素以外,因角质层的神经酰胺引起的水分保有力减少、皮肤表面的pH增加等原因引起的角质形成细胞非正常消除等,会在引起皮肤屏障功能障碍的同时成为决定皮肤疾病的严重程度的原因。并且,这些皮肤疾病的大部分集中在婴幼儿期及青少年期,引起情绪及教育方面的社会问题。
尤其,由于这些皮肤疾病伴随的难以忍受的瘙痒症引起的抓挠次数的增加进一步恶化症状。近来,查明存在于皮肤角质形成细胞中的白细胞介素及胸腺基质淋巴细胞生成素等多种细胞因子的表达增加刺激皮肤下的感觉神经而引起瘙痒症和皮炎(Mack et al.,2018,Trends Immunol..2018 Dec;39(12):980-991)。
炎症反应是在生物体或组织受到物理作用、各种化学物质的接触或者细菌感染等任意的基质的变化时引起的恢复入侵时发生损伤的部位的再生机制。
炎症成分等多种血管活性物质因外部刺激而在局部释放,这虽增加血管透过性而诱发炎症,但在某些情况下,不均衡的炎症反应会促进黏膜损伤而引起多种其他疾病(Willoughby DA.(1975)Human arthritis applied to animal models.Towards abetter therapy.Annals of the Rheumatic Disease.34,471-478.)。
作为炎症指标的促炎性细胞因子包括肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、单核细胞趋化蛋白1(MCP1)等(Yun HJ,Heo SK,Yi HS,Kim CH,Kim BW and Park SD.(2008)Anti-inflammatory effect of injinho-tang in Raw264.7Cells.Kor.J.Herbology.23(2),169-178)。
现在,特应性皮炎的治疗通常使用注射剂或软膏剂形式的皮质醇、泼尼松龙、甲基泼尼松龙、地塞米松等多种抗组胺剂或者类固醇,但无法接收充分剂量的给药。尤其,类固醇疗法会扩张毛细血管并使皮肤层变薄而使过敏反应更为严重,或者,若中断类固醇疗法,则会因类固醇戒断而使症状更为严重。
因此,在治疗并改善特应性皮炎等炎症性皮肤病方面,至今仍需要从天然资源中开发比现有使用的药物副作用少、价格低廉且有效的药物及化妆品。
龙眼肉(Longanae Arillus)为属于无患子科(Sapindaceae)的龙眼(Dimocarpuslongan或Euphoria longan)或同种的种皮,据报告,含有葡萄糖、果糖、蛋白质等,具有心脏保护、促进食欲等下效果(Chung B.S et al,Dohaehyangyakdaesajeon,youngrimsa,2ndEd.p197-198,1998)。
细叶藁本(Ligusticum tenuissimum Kitagawa)属于伞形科(Umbelliferae),为中国藁本(Ligusticum sinense Oliv)、辽藁本(Ligusticum jeholense Nakai etKitagawa)或同种的根茎或根部,据报告,藁本(Ligustici Tenuissimi Rhizoma)含有蛇床肽内酯(cnidilide)、3-丁基苯酞(phthalide),具有抗病毒效果等(Chung B.S et al,Dohae-hyangyakdaesajeon,youngrimsa,2nd Ed.P428-429,1998)。
属于远志科(Polygalaceae)的远志(Polygala tenuifolia Willd)或同种的根部远志(Polygalae radix)含有多种皂苷,据报告,具有祛痰作用、抗菌作用等(Chung B.S etal,Dohaehyangyakdaesajeon,youngrimsa,2nd Ed.P798-799,1998)。
但是,在本申请中引用的作为参考的上述文献中,没有公开任何关于对皮肤炎症疾病显出强力治疗效果的局部应用龙眼肉、藁本及远志的组合生药提取物的有关预防或改善活性的报告。
发明内容
技术问题
为了调查龙眼肉、藁本及远志的组合生药提取物的抗炎效果,本发明人集中进行了多种实验,包括:有关与皮肤炎症相关的细胞因子(酸性核糖体磷蛋白亚基(RPLPO),胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子(GM-CSF)以及白细胞介素1β)表达的抑制效果等体外实验(实验例1);通过BALB/C小鼠的特应性皮炎抑制效果等生物体内实验(实验例2);利用实验动物的有关皮肤炎症的多种细胞因子(磷酸甘油醛脱氢酶(GADPH)、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子、白细胞介素4(IL-4)、白细胞介素10(IL-10)、白细胞介素13(IL-13)、白细胞介素31(IL-31)以及白细胞介素33(IL-33))的表达抑制试验(实验例3);利用试验动物的胸腺基质淋巴细胞生成素细胞因子表达抑制试验(实验例4)。通过这些调查的结果,本发明人确认到本发明的组合生药提取物强力地抑制及改善胸腺基质淋巴细胞生成素细胞因子的表达或者治疗或改善皮肤炎症疾病,从而最终完成本发明。
技术方案
为了解决背景技术中的问题,本发明的技术方案为开发用于治疗及预防皮肤炎症疾病的新型生药剂型。
因此,本发明的目的在于,提供用于抑制胸腺基质淋巴细胞生成素细胞因子的表达或者治疗及改善皮肤炎症疾病的包含龙眼肉、藁本及远志的组合生药提取物作为有效成分的局部用药物组合物。
本申请中定义的术语“组合生药提取物”是指如下组合生药提取物,即,(a)以龙眼肉、藁本及远志的干燥重量(w/w)为基准,龙眼肉、藁本及远志的组合生药提取物的混合比为0.01-100∶0.01-100∶0.01-100重量份(w/w),优选地,混合比为0.1-50∶0.1-50∶0.1-50重量份(w/w),更优选地,混合比为0.1-10∶0.1-10∶0.1-10重量份(w/w),尤其优选地,混合比为1-5∶1-5∶1-5重量份(w/w),最优选地,混合比为1-3∶1-3∶1-3重量份(w/w);(b)以龙眼肉、藁本及远志的干燥重量(w/w)为基准,龙眼肉、藁本及远志的各提取物的组合物的混合比为0.01-100∶0.01-100∶0.01-100重量份(w/w),优选地,混合比为0.1-50∶0.1-50∶0.1-50重量份(w/w),更优选地,混合比为0.1-10∶0.1-10∶0.1-10重量份(w/w),尤其优选地,混合比为1-5∶1-5∶1-5重量份(w/w),最优选地,混合比为1-3∶1-3∶1-3重量份(w/w)。
本发明的再一目的为提供包含抑制胸腺基质淋巴细胞生成素细胞因子的量的龙眼肉、藁本及远志的组合生药提取物作为有效成分的胸腺基质淋巴细胞生成素表达抑制剂。
本申请中公开的术语“提取物”包括可以通过选自水、例如甲醇、乙醇、丙醇、丁醇等C1-C4的低级醇、丙酮、乙酸乙酯、三氯甲烷、己烷、丁二醇、丙二醇和甘油中的一种以上的溶剂提取的提取物,优选地,包括可以通过选自由水、甲醇、乙醇中的一种以上的溶剂提取的提取物,更优选地,包括可以通过选自水和10%-90%(v/v)的乙醇水溶液中的一种以上的溶剂提取的提取物,最优选地,包括可以通过选自水和20%-80%(v/v)乙醇水溶液的一种以上的溶剂提取的提取物,但不限定于此。
本申请中公开的术语“皮肤炎症疾病”包括:因老化或特应性引起的瘙痒症、特应性皮炎、银屑病等慢性复发性皮炎;接触性皮炎、脂溢性皮炎、神经性皮炎、皮肤干燥症、红斑、炎症性皮炎、银屑病或特应性疾病。优选地,包括:因老化引起的瘙痒症或因细胞因子的表达增加引起的特应性、特应性皮炎、银屑病等慢性复发性皮炎;接触性皮炎、脂溢性皮炎、神经性皮炎、皮肤干燥症、红斑、炎症性皮炎、银屑病或特应性疾病;更优选地,包括选自由因老化引起的瘙痒症或因胸腺基质淋巴细胞生成素细胞因子的表达增加引起的特应性、特应性皮炎、银屑病等慢性复发性皮炎以及接触性皮炎、脂溢性皮炎、神经性皮炎、皮肤干燥症、红斑、炎症性皮炎、银屑病或特应性疾病组成的组中的疾病。
本申请中公开的术语“抗炎”表示抑制各种炎症的所有机制,但不限定于此。
炎症为身体组织对病原体、损伤的细胞或刺激物质等有害刺激的复杂的生物学反应中的一部分,将发热、疼痛、发红、浮肿等非特异性的反应成为“炎症反应”。
炎症分为如下种类:(a)急性炎症,为身体对有害刺激的初期反应,通过血浆及白细胞(尤其是粒细胞)从血液中向受损组织的移动增加来实现,然后通过一系列的生物化学现象的传播以及局部血管系统、免疫系统及与损伤组织内的多种细胞相关的炎症反应的成熟来实现;(b)慢性炎症,为长期炎症,其特征在于,引起存在于炎症部位的细胞类型,例如单核细胞的渐进性变化,在炎症过程中同时发生组织的破坏和治愈。
通常,受损细胞的巨噬细胞释放多种细胞因子来激活T淋巴细胞,作为淋巴细胞的肥大细胞释放多种组氨引起内部屏障反应来诱导被感染细胞的炎症。因此,细胞因子的表达水平可以用作炎症反应被激活(其他方式,抗炎活性)的指标。本申请中公开的“抗炎活性”是指对皮肤炎症的抑制活性。
细胞因子是指通过趋化因子、干扰素、白细胞介素、淋巴因子及巨噬细胞、B淋巴细胞、T淋巴细胞以及肥大细胞等免疫细胞以及包括内皮细胞在内的广义的细胞生成的肿瘤坏死因子、通过成纤维细胞以及通过例如病毒等的病原体的入侵而诱发的免疫学进行来释放的包括多种基质细胞在内的所有免疫物质。
通常,细胞因子虽然只在感染初期释放,但在免疫系统非正常地被激活的情况下,会持续释放。当细胞因子以高水平持续释放一周以上的时间时,本发明人称之为“细胞因子风暴(storm)”,这是如下的生理反应,即,先天免疫体系无法调节成为细胞因子的促炎性信号传递分子而引起过度的释放,免疫细胞极度丰富地聚集于感染的部位而使炎症恶化,舒缓血管引起血管外出血,严重的甚至会引起死亡。本申请中公开的术语“细胞因子表达的抑制活性”可以解释为预防、治疗或改善细胞因子风暴。
本申请中公开的术语“细胞因子”是指与特应性皮炎等皮炎相关的多种细胞因子,具体地,包括选自由胸腺基质淋巴细胞生成素、例如粒细胞-巨噬细胞集落刺激因子、巨噬细胞集落刺激因子(M-CSF)、粒细胞集落刺激因子(G-CSF)等集落刺激因子(CSF)、例如白细胞介素-1(IL-1)、IL-4、IL-10、IL-12、IL-13、IL-31、IL-33等白细胞介素、肿瘤坏死因子α(TNF-α)、干扰素γ(IFNγ)等组成的组中的细胞因子,但不限定于此。
本发明的提取物可以通过如下优选的实施方式来制备。
在本发明的情况下,上述提取物可以通过如下方式制备。
本申请中定义的术语“龙眼肉、藁本及远志的组合生药提取物”可以通过如下步骤制备:步骤1,将龙眼肉、藁本及远志切片及清洗来用作基础提取物质;步骤2,以龙眼肉、藁本及远志的干燥重量(w/w)为基准,以0.01-100∶0.01-100∶0.01-100重量份(w/w)的混合比,优选地,以0.1-50∶0.1-50∶0.1-50重量份(w/w)的混合比,更优选地,以0.1-10∶0.1-10∶0.1-10重量份(w/w)的混合比,尤其优选地,以1-5∶1-5∶1-5重量份(w/w)的混合比,最优选地,以1-3∶1-3∶1-3重量份(w/w)范围的混合比一起混合龙眼肉、藁本及远志来获得混合物质;步骤3,将选自由水、例如甲醇、乙醇、丙醇、丁醇等C1-C4的低级醇、丙酮、乙酸乙酯、三氯甲烷、己烷、丁二醇、丙二醇和甘油组成的组中的提取溶剂,优选地,将选自由水、甲醇、乙醇组成的组中的提取溶剂,更优选地,将选自由水和10%-90%(v/v)的乙醇水溶液组成的组中的提取溶剂,最优选地,将选自水和20%-80%(v/v)乙醇水溶液组成的组中的提取溶剂,以1倍-20倍的体积(v/w)加入于混合物质,优选地,以4倍-8倍的体积(v/w)加入于混合物质;步骤4,通过在50℃至120℃的温度范围,优选地在约80℃至100℃的温度范围中进行1小时至24小时,优选地,进行2小时至12小时的热水提取、冷水提取、回流提取或超声波提取,优选地,通过热水提取的提取方法来提取各溶液;步骤5,重复上述提取过程,通过过滤、冷冻干燥或热风干燥来干燥各过滤液,优选地,通过冷冻干燥各过滤液来收集龙眼肉、藁本及远志的组合生药提取物。
本发明的还有一目的在于,提供如前所述的制备本发明的龙眼肉、藁本及远志的组合生药提取物的方法。
本发明的另一目的在于,提供用于抑制胸腺基质淋巴细胞生成素或者治疗或改善皮肤炎症疾病的包含通过上述方法制备的龙眼肉、藁本及远志的组合生药提取物作为有效成分的局部用药物组合物。
并且,根据本发明的再一实施方式,提供抑制哺乳动物的胸腺基质淋巴细胞生成素细胞因子或者治疗或改善皮肤炎症疾病的方法,包括向上述哺乳动物局部给药有效量的龙眼肉、藁本及远志的组合生药提取物及其药学上可接受的载体的步骤。
并且,根据本发明的还有一实施方式,提供将龙眼肉、藁本及远志的组合生药提取物作为有效成分来制备为用于抑制包括人类在内的哺乳动物的胸腺基质淋巴细胞生成素细胞因子或者治疗或改善皮肤炎症疾病的局部制剂的用途。
本发明的组合物还可以根据所使用的方法包含通常的载体、佐剂或稀释剂。优选地,根据用途及应用方法以适当的物质用作上述载体,但不限定于此。适当的稀释剂在Remington's Pharmaceutical Science(Mack Publishing co,Easton PA)的书面文本中列举。
以下,下述配制方法及赋形剂仅用于例示,无论从哪个方面都不是限定本发明。
本发明的组合物可以提供为包含药学上可接受的载体、佐剂或稀释剂的局部用药物组合物,例如,可以包含乳糖、蔗糖、葡萄糖、山梨糖醇、甘露醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、阿拉伯胶、海藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石粉、硬脂酸镁及矿物油。剂型还可以包含填充剂、抗凝剂、润滑剂、保湿剂、香味剂、乳化剂、防腐剂等。本发明的组合物可以通过本发明所属技术领域中广为人知的任意过程配制来在向患者给药后迅速、持续或延迟释放有效成分。
为了局部给药,本发明的提取物可以配制为包括霜剂、凝胶、面膜、喷剂溶液、乳液、软膏、洗剂、擦剂(liniment)、夜霜、溶液、悬浮液、化妆片、贴片、喷雾剂、巴布剂等局部用制剂在内的软膏及霜剂形式。
药剂学给药形式的本发明的组合物能够以其药学上可接受的盐的形式来使用,不仅可以单独或以适当的组合来使用,还可以与具有药剂学活性的化合物组合来使用。
本发明的提取物或组合物的优选剂量根据对象的状态及体重、疾病的严重程度、药物形式、给药途径及期间的不同而不同,可以通过相关从业者来选择。但是,为了获得优选的效果,通常建议以0.01g/kg-10g/kg的量局部给药本发明的提取物,优选地,以1g/kg至5g/kg(重量/天)的范围的量局部给药本发明的提取物。剂量能够以一天一种剂量或多种剂量的方式给药。从组合物的观点出发,以组合物的总重量为基准,包含0.01重量百分比至80重量百分比的本提取物,优选地,包含0.5重量百分比至50重量百分比的本提取物。
本发明人通过进行有关与皮肤炎症相关的细胞因子(酸性核糖体磷蛋白亚基、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子以及白细胞介素1β)表达的抑制效果等体外实验(实验例1);通过BALB/C小鼠的特应性皮炎抑制效果等生物体内实验(实验例2);利用实验动物的有关皮肤炎症的多种细胞因子(磷酸甘油醛脱氢酶、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子、白细胞介素4、白细胞介素10、白细胞介素13、白细胞介素31以及白细胞介素33)的表达抑制试验(实验例3);进行利用试验动物的胸腺基质淋巴细胞生成素细胞因子表达抑制试验(实验例4)确认到本发明的复合组合物具有强力的抗炎效果,因此,确认到本发明的组合提取物以局部用药剂或化妆品组合物的形式非常有用地用在皮肤炎症疾病的改善或治疗中。
本发明的又一目的在于,提供包含有效量的用于抑制胸腺基质淋巴细胞生成素细胞因子或者治疗或改善皮肤炎症疾病的龙眼肉、藁本及远志的组合生药提取物作为有效成分的化妆品组合物。
以组合物的总重量为基准,本发明的化妆品组合物包含0.001重量百分比-40重量百分比的本发明的组合物,优选地,包含0.01重量百分比-10重量百分比的本发明的组合物。其他成分可以为本发明所属技术领域中公知的通常的化妆品组合物成分的混合物。
包含上述组成的化妆品的剂型可以制备为例如润肤液、柔肤水、爽肤水、收敛剂、润肤乳、乳液、保湿乳液、滋养乳液、按摩霜、滋养霜、保湿霜、护手霜、粉底、精华、滋养精华、面膜、洁面泡沫、洁面乳液、卸妆面霜、润肤露、沐浴露、润发乳、美容液等的任意形式。
以下,下述配制方法及赋形剂仅用于例示,无论从哪个方面都不是限定本发明。
本发明的化妆品组合物还可以包含选自由水溶性维生素、脂溶性维生素、肽聚合物、多糖类聚合物、鞘脂及海藻提取物组成的组中的追加添加剂。
优选的水溶性维生素可以与化妆品混合,优选地,多种维生素可以为例如维生素B1、B2、维生素B6、吡哆醇、盐酸吡哆醇、维生素B12、泛酸、烟酸、烟酰胺、叶酸、维生素C、维生素H等、它们的盐,例如维生素盐酸盐、抗坏血酸钠盐等或它们的衍生物,例如抗坏血酸-2-膦酸钠盐、抗坏血酸-2-膦酸镁盐,这些维生素可以通过微生物转化法、从微生物培养物中纯化的方法、生物酶法或化学合成法等通常的方法获得。
优选的脂溶性维生素可以与化妆品混合,优选地,本发明的实施例使用的脂溶性维生素包括多种维生素,例如维生素A、维生素D2、维生素D3、维生素E(dl-a-生育酚、d-a-生育酚、d-d-生育酚)及他们的衍生物,例如抗坏血酸棕榈酸酯、抗坏血酸硬脂酸酯、抗坏血酸二棕榈酸、乙酸-dl-a-生育酚、烟酸dl-a-生育酚维生素E、dl-泛醇、D-泛醇、泛酸乙醚等,可以通过微生物转化法、从微生物中纯化的方法、生物酶法或化学合成法等通常的方法来获得。
优选的肽聚合物可以与化妆品混合,优选地,本发明的实施例中使用的肽聚合物包括胶原蛋白、水解胶原蛋白、明胶、弹性蛋白、水解凝胶、角蛋白等。
优选的多糖类聚合物可以与化妆品混合,优选地,上述多糖类聚合物为羟乙基纤维素、黄原胶、透明质酸钠、硫酸软骨素或它们的盐(钠盐等Na)。例如,可以从哺乳动物或鱼类中纯化硫酸软骨素来使用。
优选的鞘脂可以与化妆品混合,优选地,上述鞘脂为神经酰胺、植物鞘氨醇、鞘脂多糖等。鞘脂可以通过通常的方法从哺乳动物、鱼类、贝壳类、酵母或植物等中获得。
优选的海藻提取物(seaweed extract)可以与化妆品混合,优选地,上述海藻提取物为褐藻类、红枣类、绿藻类等的提取物或从它们分离纯化的角叉菜胶、海藻酸、海藻酸钠、海藻酸钾等。海藻类提取物(algae extract)从海藻中纯化来获得。
根据需要,可以将上述必需成分与通常的化妆品组合物所使用的其他成分一同组合为本发明的化妆品组合物。
优选地,上述记载的其他成分可以包括油成分、保湿剂、软化剂、表面活性剂、有机或无机染料、有机粉体、紫外线吸收剂、保存剂、防腐剂、抗氧化剂、植物提取物、pH调节剂、醇类、颜料、香水、制冷剂、血液循环改善剂(circulator)、止汗剂(antihidrotic)、蒸馏水等。
优选地,油成分可以包括酯油、碳氢油、硅油、氟化物油、动物油、植物油等。
优选地,上述记载的酯油可以包括甘油三-2-乙基己酸,十六烷基-2-乙基己酸、肉豆蔻酸异丙酯、肉豆蔻酸丁酯、棕榈酸异丙酯、硬脂酸乙酯、棕榈酸辛酯、异十六烷基异硬脂酸、硬脂酸丁酯、亚油酸乙酯、异丙基亚油酸、乙基油酸、异十六烷基肉豆蔻酸、异硬脂基肉豆蔻酸、异硬脂基棕榈酸、辛基十二烷基肉豆蔻酸,异十六烷基异硬脂酸、癸二酸二乙酯、己二酸异丙酯、异烷基新戊酸、甘油三辛酯、甘油三癸酯、三甲基丙烷三-2-乙基乙酸、三甲基丙烷三异硬脂酸、季戊四醇四-2-乙基乙酸、鲸蜡基辛酸、癸基月桂酸、己基月桂酸、癸基肉豆蔻酸、肉豆蔻基肉豆蔻酸、鲸蜡基肉豆蔻酸、硬脂基硬脂酸、癸基油酸、鲸蜡基蓖麻油酸、异硬脂基月桂酸、异十三烷基肉豆蔻酸、异鲸蜡基棕榈酸、辛基硬脂酸、异鲸蜡基硬脂酸、异癸基油酸、辛基十二烷基油酸、辛基十二烷基亚油酸、异丙基异硬脂酸、鲸蜡硬脂基2-乙基酸、硬脂基2-乙基酸、己基异硬脂酸、乙二醇二辛酸、乙二醇二油酸、丙二醇二癸酸、丙二醇二辛脂、丙二醇二癸酯、丙二醇二辛酸、新戊二醇二癸酸、新戊二醇二辛酸、甘油三辛酸、甘油三十一烷酸、甘油三棕榈酸、甘油三异硬脂酸、辛基十二烷基新戊酸、异硬脂基辛酸、辛基异壬酸、十六烷基新癸酸、辛基十二烷基新癸酸、异十六烷基异硬脂酸、异硬脂基异硬脂山、辛基癸基异硬脂酸,聚甘油齐墩果醇酸酯、聚甘油异硬脂酸酯、三异十六烷基柠檬酸、三异烷基柠檬酸、三异辛基柠檬酸、月桂基乳酸、肉豆蔻基乳酸、鲸蜡基乳酸、辛基癸基乳酸、柠檬酸三乙酯、乙酰基柠檬酸三乙酯、乙酰基柠檬酸三丁酯、三辛基柠檬酸、二异硬脂基马来酸、二2-乙基己基羟基硬脂酸、2-乙基己基琥珀酸、二异丁基己二酸、癸二酸二异丙酯、癸二酸二辛酯、胆甾醇硬脂酸、胆甾醇异硬脂酸、胆甾醇羟基硬脂酸、胆甾醇羟基硬脂酸、胆固醇油酸、二氢胆固醇油酸、植物甾醇异硬脂酸、植物甾醇油酸、异十六烷基12-硬脂酰羟基硬脂酸、硬脂基12-硬脂酰羟基硬脂酸、异硬脂酰12-硬脂酰羟基硬脂酸。
优选地,上述记载的碳氢油包括角鲨烯、液体石蜡、α-烯烃低聚物、异链烷烃、地蜡、石蜡、液体异石蜡、聚丁烯、微晶蜡、凡士林。
优选地,上述记载的硅油可以包括聚甲基硅油、甲基苯基硅氧烷、甲基环聚硅氧烷、八甲基聚硅氧烷、十甲基聚硅氧烷、十二甲基环硅氧烷、二甲基硅氧烷-甲基十六烷氧基硅氧烷共聚物、二甲基硅氧烷-甲基十八烷氧基硅氧烷共聚物、烷基改性硅油、氨基改性硅油等。
优选地,上述记载的氟化物油可以包括全氟聚醚等。
优选地,动物油或植物油可以包括鳄梨油、杏仁油、橄榄油、芝麻油、米糠油、红花油、大豆油、玉米油、菜籽油、苦杏仁油、棕榈仁油、棕榈油、蓖麻油、葵花籽油、果籽油、棉籽油、椰子油(coconut palm oil)、石栗油(cucui nut oil)、小麦胚芽油(wheat embryo budoil)、米胚芽油、乳木果油、月见草油、澳洲坚果油(marker daymia nut oil)、白芒花籽油(medo home oil)、蛋黄油、羊毛脂、大麻籽油、貂油、甜橙油(orange ruppy oil)、荷荷巴油、大麻蜡、液体羊毛脂、固体蓖麻蜡等。
优选地,保湿剂可以包括水溶性低分子保湿剂、亲油性低分子保湿剂、水溶性聚合物及脂溶性聚合物等。
具体地,优选的水溶性低分子保湿剂可以包括丝氨酸、谷氨酰胺、山梨糖醇、甘露醇、吡咯烷酮羧酸钠、甘油、丙二醇、1,3-丁二醇、乙二醇、聚乙二醇(聚合度>2)、聚丙二醇(聚合度>2)、乳酸、乳酸盐等。
优选的脂溶性低分子保湿剂可以包括胆固醇、胆固醇酯等。
优选的水溶性聚合物可以包括羧乙基聚合物、聚天冬氨酸盐、黄芪胶、黄原胶、羟甲基纤维素(HMC)、羟乙基纤维素(HEC)、羟丙基纤维素(HPC)、羧甲基纤维素、水溶性甲壳素、壳聚糖、糊精等。
优选的脂溶性聚合物可以包括聚乙烯吡咯烷酮-二十烯共聚物、聚乙烯吡咯烷酮-十六烯共聚物、硝酸纤维素、糊精脂肪酸酯、有机硅聚合物等。
优选的软化剂可以包括长链酰基谷氨酸胆固醇酯、胆固醇羟基硬脂酸、12-羟基硬脂酸、松香酸(rogic acid)、羊毛脂脂肪酸胆固醇酯等。
优选的表面活性剂可以包括非离子型表面活性剂、阴离子型表面活性剂、阳离子型表面活性剂、两性表面活性剂等。
具体地,优选的非离子型表面活性剂可以包括自乳化单硬脂酸甘油酯、脂肪酸丙二醇酯、脂肪酸甘油酯、局脂肪酸甘油酯、山梨糖醇酐脂肪酸酯、聚氧乙烯(POE)山梨糖醇酐脂肪酸酯、聚氧乙烯甘油脂肪酸酯、聚氧乙烯烷基醚、聚氧乙烯脂肪酸酯、聚氧乙烯固体蓖麻油、聚氧乙烯蓖麻油、聚氧乙烯-聚烯烃塑性体(POE-POP)共聚物、聚氧乙烯-聚烯烃塑性体烷基醚、聚醚改性有机硅、月桂烷醇酰胺、烷基氧化胺、氢化大豆磷脂等。
优选的阴离子型表面活性剂可以包括脂肪酸皂、α-酰基磺酸盐、烷基磺酸盐、烷基芳基磺酸盐、烷基萘磺酸盐、烷基磺酸盐、聚氧乙烯烷基醚硫酸盐、烷基酰胺硫酸盐、烷基磷酸盐、聚氧乙烯烷基磷酸盐、烷基酰胺磷酸盐、烷酰基烷基牛磺酸盐、N-酰基-氨基酸盐、聚氧乙烯烷基醚羧酸盐、烷基磺基琥珀酸盐、烷基磺基乙酸盐、酰化可水解胶原肽盐、全氟烷基磷酸酯等。
优选的阳离子型表面活性剂可以包括:烷基三甲基氯化铵、硬脂基三甲基氯化铵、硬脂基三甲基溴化铵、鲸蜡硬脂基三甲基氯化铵、二硬脂基二甲基氯化铵、硬脂基二甲基苄基氯化铵、山嵛基三甲基溴化铵、苯扎氯铵、硬脂酸二乙氨基乙酯、硬脂酸二甲氨基丙酰胺、羊毛脂衍生物季铵盐等。
优选的两性表面活性剂可以包括羧基甜菜碱型、酰胺甜菜碱型、羟基磺基甜菜碱型、磷酸甜菜碱型、氨基羧酸、咪唑啉衍生物型、酰胺胺型等。
优选的有机及无机染料可以包括:无机染料,硅酸、硅酸酐、硅酸镁、滑石、绢云母、云母、高岭土、孟加拉、克洛伊、膨润土、钛膜云母、氯氧化铋、氧化锆、氧化镁、氧化锌、氧化钛、氧化铝、硫酸钙、硫酸钡、硫酸镁、碳酸钙、碳酸镁、氧化亚铁、氧化铬、氢氧化铬、炉甘石、炭黑以及它们的复合物;有机染料,聚酰胺、聚酯、聚丙烯、聚苯乙烯、聚氨酯、乙烯基树脂、尿素树脂、酚醛树脂、氟树脂、硅树脂(silicone resin)、丙烯酸树脂、三聚氰胺树脂、环氧树脂、聚碳酸酯树脂、二乙烯基苯-苯乙烯共聚物、蚕丝蛋白、纤维素、CI颜料黄、CI颜料橙以及它们的复合物。
优选的有机粉体可以包括:金属皂,例如茶酸钙;烷基磷酸金属盐(alkylphosphonate metal salt),例如鲸蜡酸钠、月桂酸锌、月桂酸钙;酰基氨基酸多价金属盐,例如N-月桂酰-b-丙氨酸钙、N-月桂酰-b-丙氨酸锌、N-月桂酰-甘氨酸钙等;酰胺磺酸多价金属盐,例如N-月桂酰牛磺酸钙、N-棕榈酰牛磺酸钙等;N-酰基碱性氨基酸,例如Nε-月桂酰-L-赖氨酸、Nε-棕榈酰赖氨酸、Nα-棕榈酰鸟氨酸、Nα-月桂酰精氨酸、氢化羊毛脂脂肪酸酰基精氨酸等;N-酰基多肽,例如N-月桂酰甘氨酸;α-氨基脂肪酸,例如α-氨基辛酸、α-氨基月桂酸等;以及聚乙烯、聚丙烯、尼龙、聚甲基丙烯酸甲酯、聚苯乙烯、二乙烯基苯-苯乙烯共聚物、四氟化乙烯等。
优选的紫外线吸收剂可以包括对氨基苯甲酸、对氨基苯甲酸乙酯、对氨基苯甲酸戊酯、对氨基苯甲酸辛酯、水杨酸乙二醇酯、水杨酸苯酯、水杨酸辛酯、水杨酸苄酯、水杨酸丁苯酯、水杨酸高薄荷酯、肉桂酸苄酯、对甲氧基肉桂酸酯-2-乙氧基乙基酯、对甲氧基肉桂酸酯辛酯、二对甲氧基肉桂酸酯单-2-乙基甘油酯、对甲氧基肉桂酸酯异丙基酯、二异丙基-二异丙基肉桂酸酯混合物,尿刊酸,尿烷酸乙酯,羟基甲氧基二苯甲酮,羟基甲氧基二苯甲酮磺酸及其盐,二羟基甲氧基二苯甲酮,二羟基甲氧基二苯甲酮二磺酸钠(dihydroxymethoxy benzophenone disulfonate Na)、二羟基二苯甲酮、四羟基二苯甲酮、4-叔丁基-4'-甲氧基二苯甲酰甲烷、2,4,6-三苯胺基-p-(羧基-2'-乙基己基-1'-oxy)-1,3,5-三嗪、2-(2-羟基-5-甲基苯基)苯并三唑等。
优选的保存剂可以包括扁柏醇、三盐酸、三氯羟基二苯醚、葡萄糖醛酸氯己定、苯氧基乙醇、间苯二酚、异丙基甲基苯酚、薁、水杨酸、苯锂硫锌、盐酸贝沙氯铵、感光剂301、单硝基愈创木酚钠、十一烯酸等。
优选的抗氧化剂可以包括丁基羟基苯甲醚、没食子酸丙酯、依山梨酸等。
优选的pH调节剂可以包括柠檬酸、柠檬酸钠、苹果酸、苹果酸钠、富马酸、富马酸钠、琥珀酸、琥珀酸钠、磷酸氢钠等。
优选的醇类可以包括鲸蜡醇等。
并且,在本发明的目的及效果范围内,可以向上述成分添加的其他成分及其量不受限制,优选地,其他成分的含量为组合物总含量中的0.01%至5%,更优选地,为0.01%至3%的范围。
本发明的化妆品组合物可以变形为溶液、乳液、粘附性混合物等。
水溶性维生素、脂溶性维生素、肽聚合物、多糖类聚合物、鞘脂、海藻提取物等上述成分以及根据需要添加的除上述成分以外的添加成分可以通过文献中记载的通常的方法来获取(Matsumoto Mithio;Manual for the development of transdermal appliedpreparation.Seisi Press,1st Ed.,1985)。
本发明的化合物无毒副作用,因此可以安全地使用。
本发明所属技术领域的普通技术人员应该明白的是,在不脱离本发明的精神或范围的情况下,本发明的组合物、用途及配制可以进行多种变更及变形。
通过下述实施例更为具体地说明本发明。但是,无论在任何方面都不应理解为本发明限定于这些实施例。
有益效果
如上所述,本发明人通过进行有关与皮肤炎症相关的细胞因子(酸性核糖体磷蛋白亚基、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子以及白细胞介素1β)表达的抑制效果等体外实验(实验例1);通过BALB/C小鼠的特应性皮炎抑制效果等生物体内实验(实验例2);利用试验动物的有关皮肤炎症的多种细胞因子(磷酸甘油醛脱氢酶、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子、白细胞介素4、白细胞介素10、白细胞介素13、白细胞介素31以及白细胞介素33)的表达抑制试验(实验例3);利用试验动物的胸腺基质淋巴细胞生成素细胞因子表达抑制试验(实验例4)确认到本发明的复合组合物具有强力的抗炎效果,因此,确认到本发明的组合提取物以局部用药剂或化妆品组合物的形式非常有用地用在皮肤炎症疾病的改善或治疗中。
附图说明
图1示出处理试验试样(WIN)、地塞米松(DEX)及蒸馏水(DIW)的皮炎诱发背部皮肤的病变。
图2示出在处理试验试样(WIN)、地塞米松(DEX)及蒸馏水(DIW)的皮炎诱发背部皮肤病变上进行苏木精伊红染色(H&E)和甲苯胺蓝(TB)染色试验的结果。
具体实施方式
本发明所属技术领域的普通技术人员应该明白的是,在不脱离本发明的精神或范围的情况下,本发明的组合物、用途及配制可以进行多种变更及变形。
通过下述实施例更为具体地说明本发明。但是,无论在任何方面都不应理解为本发明限定于这些实施例。
实施例
在不限制本发明的范围的情况下,下述实施例及实验例用于追加例示本发明。
实施例1.制备本发明的组合提取物(1)
细细切割20g的干燥龙眼肉(Buyoung Yakup Co.Ltd.公司)、20g的干燥藁本(Buyoung Yakup Co.Ltd.公司)以及20g的干燥远志(Buyoung Yakup Co.Ltd.公司)后,与6倍体积(v/w)的20%的乙醇水溶液混合,在90±5℃的温度下回流提取该混合物3天。通过过滤纸(孔隙大小:小于10μm)过滤提取物去除残余物后,使用4倍体积(v/w)的20%的乙醇水溶液再提取剩余的残余物两次后,通过过滤纸(孔隙大小:小于10μm)过滤提取物。
将收集的提取物混合在一起后在真空下进行浓缩(16白利糖度-21白利糖度)来获得浓缩的提取物。通过冷冻干燥过程干燥浓缩的提取物后,通过粉碎(小于50目)获得20.5g的本发明的组合提取物(1)(以下表示为“WIN-1001X”)(干燥粉末,收率33.4%)。
实施例2至实施例6.制备本发明的组合提取物(2)至本发明的组合提取物(6)
除使用不同混合比和不同溶剂以外,以与实施例1相同的过程来获得龙眼肉(LA)、藁本(LT)及远志(PR)的多种本发明的组合提取物,即,本发明的组合提取物(2)至本发明的组合提取物(6),在下述实验中使用他们作为试验试样。
表1
各种组合提取物
实验例1.对细胞因子表达的抑制效果(体外).
为了确定本发明提取物的抗炎活性,根据文献记载的过程进行利用HaCaT细胞的下述细胞因子表达抑制试验(Jeong et al.,2019,J.Invest.Dermatol.,May;139(5):pp1098-1109)。
在包含10%的胎牛血清、100单位/ml的青霉素、100μg/ml的链霉素(D6429,Sigma-Aldrich Co.Ltd公司)的杜氏改良伊戈尔培养基(DMEM)中接种HaCaT细胞(人类上皮角质细胞,300493,CLS公司)后,在保持最佳湿度(85%-95%)及5%的CO2大气气氛的条件下在恒温箱(HERA cell 150i,Thermo Fisher Scientific Co.Ltd.公司)中进行培养。
为了进行基因表达试验,将培养的细胞移到12个孔中后,使用50ng/ml的肿瘤坏死因子α(RC214-12,Biobasic Co.Ltd公司)处理1小时来诱导炎症反应。将地塞米松(200nM,阳性对照组,“DEX”,D4902,Sigma-Aldrich Co.Ltd.公司)及蒸馏水(阴性对照组,“DIW”)用作比较对照组。
诱导炎症1小时后,使用相同的培养基处理1μg/ml的实施例中制备的本发明的提取物后培养1小时。培养后,从细胞中提取核糖核酸(RNA)(FATRR-001,Favorgen公司)后,使用互补脱氧核糖核酸(cDNA)合成试剂盒(RRO36A,TAKARA公司)从核糖核酸合成互补脱氧核糖核酸。使用合成的互补脱氧核糖核酸与Sybrgreen试剂盒(RT500M,Enzynomics公司)进行聚合反应后,如表2所示,利用与皮肤炎症有关的多种细胞因子的相关引物(RPLPO、TSLP、GM-CSF以及IL-1β)进行实时聚合酶链式反应(PCR)。
表2
实时聚合酶链式反应(RT-PCR)方法中使用的引物
实时聚合酶链式反应的定量结果如表3所示,通过表3可知,与处理蒸馏水(DIW)的阴性对照组相比,处理本发明的提取物的试验试样组迅速抑制了与皮肤炎症相关的多种细胞因子的表达水平,确认到试验试样对与皮肤炎症相关的多种细胞因子的表达的抑制活性与处理地塞米松(DEX)的阳性对照组相同。
因此,确认实施例1至实施例6中制备的各种本发明的组合提取物对皮肤炎症具有强力的抑制效果。
表3
细胞因子表达抑制效果
实验例2.特应性皮炎抑制效果(生物体内)
为了确认本发明提取物的特应性皮炎抑制效果,根据参考文献(Li et al.,Drug.Des.Devel.Ther.,2016,Feb.19;10:781-191)中记载的方法进行利用小鼠的动物模型试验。
将100μl的0.15%(w/v)的2,4-二硝基氟苯(DNFB,D1529,Sigma-Aldrich Co.Ltd.公司)涂敷于6周龄的BALB/C雌性小鼠(DBL Co.,Ltd.公司,仁川(Incheon),韩国(Korea))的腹腔,去除背部中央的毛后,从第7天到第16天每隔三天涂敷来诱导皮肤炎症。从2,4-二硝基氟苯的初期治疗日的第7天开始到第16天,不仅涂敷试验试样(10mg/ml的本发明的提取物WIN-1001X,实施例中制备),还使用地塞米松(200μM,阳性对照组“DEX”,D4902,Sigma-Aldrich Co.Ltd.公司)以及蒸馏水(阴性对照组,“DIW”)进行涂敷来作为比较对照组。
为了比较试验结果,根据上述过程,除使用丙酮替代2,4-二硝基氟苯,使用蒸馏水替代试验试样以外,进行相同的试验。
通过图1可知,在处理试样的第16天,小鼠背部皮肤发生皮炎,皮炎的严重程度分为4种标准,即,(i)红斑/出血,(ii)浮肿,(iii)擦刮伤/充血,(iv)疤痕/干燥,在总12分的情况下,分为4个分数,即,0分(无症状),1分(单纯皮炎),2分,(平均皮炎),4分(重症皮炎)。
通过图1及表4可知,与阴性对照组相比,本发明组合提取物表现出强力的皮炎改善效果,确认到表现出与处理地塞米松的阳性对照组相似的效果。
表4
皮炎改善效果
并且,进行了有关试验动物的皮肤组织的组织学分析。
具体地,在处理试样第16天以及固定后的第12小时在保持4℃的振荡机(CR300,FINEPCR公司)中将发生皮炎的背部皮肤组织固定于4%(w/v)的多聚甲醛溶液(P6148,Sigma-Aldrich公司)后,将脱水的透明组织包埋于石蜡中,通过切割(5μm的宽度)制备组织切片。
使用用于染色皮肤表皮层的H&E染色剂(苏木精(S3309,DAKO公司)和伊红(109844,Millipore公司))或用于染色渗透炎症病变的肥大细胞的甲苯胺蓝(TB,185426,Sigma-Aldrich公司)染色组织切片,使用显微镜(EVOS XL.,Life Technologies公司)观察结果。
通过图2及表5-表6可知,与处理蒸馏水的阴性对照组相比,处理本发明组合提取物的试验组的皮肤表皮层的宽度显著减小,渗透到炎症病变的肥大细胞的数量减少,确认到其效果与处理地塞米松的阳性对照组相似。
表5
有关表皮的相对面积的效果(倍)
表6
有关肥大细胞的相对数量的效果(倍)
实验例3.细胞因子表达抑制效果(生物体内)
为了确认本发明提取物的抗炎活性,根据文献(Li et al.,Drug.Des.Devel.Ther.,2016,Feb 19;10:781-191)中记载的过程进行利用试验动物的下述细胞因子表达抑制试验。
在处理2,4-二硝基氟苯后的第16天,从实验例2中制备的小鼠背部皮肤中提取核糖核酸(FATRR-001,Favorgan公司),利用互补脱氧核糖核酸合成试剂盒(RRO36A,TAKARA公司)从核糖核酸合成互补脱氧核糖核酸。使用合成的互补脱氧核糖核酸与Sybrgreen试剂盒(RT500M,Enzynomics公司)进行聚合反应后,如表7所示,利用与皮肤炎症有关的多种细胞因子(磷酸甘油醛脱氢酶、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子、白细胞介素4、白细胞介素10、白细胞介素13、白细胞介素31以及白细胞介素33)的相关引物进行实时聚合酶链式反应(PCR)。
表7
实时聚合酶链式反应的定量结果如表8所示,通过表3可知,与处理蒸馏水(DIW)的阴性对照组相比,处理本发明的提取物的试验试样组迅速抑制了与皮肤炎症相关的多种细胞因子的表达水平,确认到试验试样对与皮肤炎症相关的多种细胞因子的表达的抑制活性与处理地塞米松(DEX)的阳性对照组相同。
因此,确认实施例中制备的各种本发明的组合提取物对皮肤炎症具有强力的抑制效果。
表8
细胞因子表达抑制效果
实验例4.胸腺基质淋巴细胞生成素细胞因子表达抑制效果(生物体内)
为了确认本发明提取物的抗炎活性,根据(Li et al.,Drug.Des.Devel.Ther.,2016,Feb 19;10:781-191)中记载的过程,进行利用试验动物的下述胸腺基质淋巴细胞生成素细胞因子表达抑制试验。
在处理2,4-二硝基氟苯后的第16天,从实验例2中制备的小鼠背部皮肤中提取核糖核酸(FATRR-001,Favorgan公司),利用互补脱氧核糖核酸合成试剂盒(RRO36A,TAKARA公司)从核糖核酸合成互补脱氧核糖核酸。使用合成的互补脱氧核糖核酸与Sybrgreen试剂盒(RT500M,Enzynomics公司)进行聚合反应后,使用与表7所示的与皮肤炎症相关的胸腺基质淋巴细胞生成素细胞因子相关的引物进行实时聚合酶链式反应。
通过示出实时聚合酶链式反应的定量结果的表9可知,与处理蒸馏水的阴性对照组相比,处理本发明提取物的试验试样组迅速抑制了与皮肤炎症相关的胸腺基质淋巴细胞生成素细胞因子的表达水平,确认到试验试样组对胸腺基质淋巴细胞生成素细胞因子的表达的抑制活性与处理地塞米松的阳性对照组相同。
因此,确认实施例中制备的本发明组合提取物具有对胸腺基质淋巴细胞生成素细胞因子表达的强力抑制效果。
表9
胸腺基质淋巴细胞生成素细胞因子表达抑制效果
统计分析
通过实验中获得的实验结果计算平均值与标准误。利用t-检验分析显著性检验,显著性(P值)以P≤0.05=*、P≤0.01=**及P≤0.001=***来表示。
本发明的方法
以下,说明赋形剂的配制及种类,但本发明不限定于此。代表性的制备例如下。
制备护肤液
1.00%的实施例的提取物(WIN-1001X)
3.00%的甘油
1.00%的乙醇
0.10%的丙二醇
微量香料
100%以下的蒸馏水
根据通常的护肤液制备方法溶解有效成分来制备皮肤制剂。
制备润肤乳
3.00%的实施例的提取物(WIN-1002X)
1.00%的L-抗坏血酸-2-磷酸镁
1.00%的可溶性胶原蛋白(1%的溶液)
0.10%的柠檬酸钠
3.00%的1,3-丁二醇
100%以下的蒸馏水
根据通常的润肤乳制备方法溶解有效成分来制备润肤乳。
制备霜剂
3.00%的实施例的提取物(WIN-1003X)
2.00%的聚乙二醇单硬脂酸酯
1.00%的单硬脂酸甘油酯
4.00%的鲸蜡醇
6.00%的角鲨烯
6.00%的三2-甘油乙基己酸酯
1.00%的鞘糖脂
3.00%的1,3-丁二醇
100%以下的蒸馏水
根据通常的霜剂制备方法溶解有效成分来制备霜剂。
制备面膜
5.00%实施例的提取物(WIN-1004X)
13.00%聚乙烯醇
1.00%的L-抗坏血酸-2-磷酸镁
1.00%的月桂酰羟基脯氨酸
2.00%的可溶性胶原蛋白(1%的溶液)
3.00%的1,3-丁二醇
5.00%的乙醇
100%以下的蒸馏水
20g的糖
20g的果糖
适量柠檬香料
100ml的蒸馏水
根据通常的面膜制备方法溶解有效成分来制备面膜。
制备美容液
2.00%的实施例的提取物(WIN-1005X)
12.00%的羟乙烯纤维素(2%的溶液)
2.00%的黄原胶(2%的溶液)
3.00%的1,3-丁二醇
4.00%浓度的甘油
5.00%的透明质酸钠
100ml的蒸馏水
根据通常的美容液制备方法溶解有效成分来制备美容液制剂。
因此,显而易见的是,说明的本发明可以变形为多种方式。这些变形不能视为脱离本发明的精神及范围,对于本发明所属技术领域的普通人员来说显而易见的是,所有变形都包括在随附的发明要求保护范围内。
根据本发明的上述记载,应该明了的是,相同的内容可以变形为多种方式。这些变形不能视为脱离本发明的精神及范围,对于本发明所属技术领域的普通人员来说显而易见的是,所有变形都包括在随附的发明要求保护范围内。
产业上的可应用性
如在本发明中说明的,本发明提供包含龙眼肉、藁本及远志的组合生药提取物的局部用组合物及化妆品组合物,本发明人通过进行有关与皮肤炎症相关的细胞因子(酸性核糖体磷蛋白亚基、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子以及白细胞介素1β)表达的抑制效果等体外实验(实验例1);通过BALB/C小鼠的特应性皮炎抑制效果等生物体内实验(实验例2);利用试验动物的有关皮肤炎症的多种细胞因子(磷酸甘油醛脱氢酶、胸腺基质淋巴细胞生成素、粒细胞-巨噬细胞集落刺激因子、白细胞介素4、白细胞介素10、白细胞介素13、白细胞介素31以及白细胞介素33)的表达抑制试验(实验例3);利用试验动物的胸腺基质淋巴细胞生成素细胞因子表达抑制试验(实验例4)验证本发明的复合组合物具有强力的抗炎效果,因此,确认到本发明的组合提取物以局部用药剂或化妆品组合物的形式非常有用地用在皮肤炎症疾病的改善或治疗中。
<110> 朴玉南
(株)美迪合伙来音
<120> 抑制TSLP及治疗或改善皮肤炎症疾病的包括含龙眼肉组合生药提取物的局部用组合物及其用途
<130> DIF/2021-02-002/EK
<150> KR 10-2020-0031482
<151> 2020-03-13
<150> KR 10-2021-0022675
<151> 2020-02-19
<160> 26
<170> KoPatentIn 3.0
<210> 1
<211> 19
<212> DNA
<213> 智人
<400> 1
agcccagaac actggtctc 19
<210> 2
<211> 21
<212> DNA
<213> 智人
<400> 2
actcaggatt tcaatggtgc c 21
<210> 3
<211> 23
<212> DNA
<213> 智人
<400> 3
tatgagtggg accaaaagta ccg 23
<210> 4
<211> 22
<212> DNA
<213> 智人
<400> 4
gggattgaag gttaggctct gg 22
<210> 5
<211> 22
<212> DNA
<213> 智人
<400> 5
tcctgaacct gagtagagac ac 22
<210> 6
<211> 19
<212> DNA
<213> 智人
<400> 6
tgctgcttgt agtggctgg 19
<210> 7
<211> 20
<212> DNA
<213> 智人
<400> 7
ctccagggac aggatatgga 20
<210> 8
<211> 20
<212> DNA
<213> 智人
<400> 8
tctttcaaca cgcaggacag 20
<210> 9
<211> 21
<212> DNA
<213> 小鼠
<400> 9
aggtcggtgt gaacggattt g 21
<210> 10
<211> 23
<212> DNA
<213> 小鼠
<400> 10
tgtagaccat gtagttgagg tca 23
<210> 11
<211> 22
<212> DNA
<213> 小鼠
<400> 11
agcttgtctc ctgaaaatcg ag 22
<210> 12
<211> 22
<212> DNA
<213> 小鼠
<400> 12
aggtttgatt caggcagatg tt 22
<210> 13
<211> 20
<212> DNA
<213> 小鼠
<400> 13
agggtctacg gggcaatttc 20
<210> 14
<211> 21
<212> DNA
<213> 小鼠
<400> 14
tcacagtccg tttccggagt t 21
<210> 15
<211> 20
<212> DNA
<213> 小鼠
<400> 15
ggtctcaacc cccagctagt 20
<210> 16
<211> 23
<212> DNA
<213> 小鼠
<400> 16
gccgatgatc tctctcaagt gat 23
<210> 17
<211> 21
<212> DNA
<213> 小鼠
<400> 17
gctcttactg actggcatga g 21
<210> 18
<211> 20
<212> DNA
<213> 小鼠
<400> 18
cgcagctcta ggagcatgtg 20
<210> 19
<211> 19
<212> DNA
<213> 小鼠
<400> 19
cctggctctt gcttgcctt 19
<210> 20
<211> 21
<212> DNA
<213> 小鼠
<400> 20
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<210> 21
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<400> 21
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<210> 22
<211> 21
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<400> 22
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<210> 23
<211> 22
<212> DNA
<213> 小鼠
<400> 23
tcagcagacg aatcaataca gc 22
<210> 24
<211> 22
<212> DNA
<213> 小鼠
<400> 24
tcgctcaaca cttggacttt ct 22
<210> 25
<211> 22
<212> DNA
<213> 小鼠
<400> 25
gctgcagaag ggagaaatca cg 22
<210> 26
<211> 30
<212> DNA
<213> 小鼠
<400> 26
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Claims (11)
1.一种局部用药物组合物,其特征在于,包含龙眼肉、藁本及远志的组合生药提取物作为抑制胸腺基质淋巴细胞生成素细胞因子的表达或者治疗及改善皮肤炎症疾病的有效成分。
2.根据权利要求1所述的局部用药物组合物,其特征在于,
在上述组合生药提取物中,
(a)以龙眼肉、藁本及远志的干燥重量(w/w)为基准,龙眼肉、藁本及远志的组合生药提取物的混合比在0.01-100∶0.01-100∶0.01-100重量份(w/w)范围内;或者
(b)以龙眼肉、藁本及远志的干燥重量(w/w)为基准,龙眼肉、藁本及远志的各提取物的组合物的混合比在0.01-100∶0.01-100∶0.01-100重量份(w/w)范围内。
3.根据权利要求1所述的局部用药物组合物,其特征在于,上述提取物使用选自水、包括甲醇、乙醇、丙醇、丁醇在内的C1-C4低级烷醇、丙酮、乙酸乙酯、三氯甲烷、己烷、丁二醇、丙二醇或甘油中的一种以上的溶剂提取。
4.根据权利要求1所述的局部用药物组合物,其特征在于,上述皮肤炎症疾病为选自由因老化或特应性引起的瘙痒症、包括特应性皮炎、银屑病在内的慢性复发性皮炎、接触性皮炎、脂溢性皮炎、神经性皮炎、皮肤干燥症、红斑、炎性皮炎、银屑病或特应性疾病组成的组中的疾病。
5.一种抑制胸腺基质淋巴细胞生成素细胞细胞因子或者治疗或改善哺乳动物的皮肤炎症疾病的方法,其特征在于,包括向上述哺乳动物局部给药有效量的龙眼肉、藁本及远志的组合生药提取物及其药学上可接受的载体的步骤。
6.一种龙眼肉、藁本及远志的组合生药提取物的用途,其特征在于,用来制备以上述龙眼肉、藁本及远志的组合生药提取物为有效成分的用于抑制胸腺基质淋巴细胞生成素细胞因子或者治疗或改善包括人类在内的哺乳动物的皮肤炎症疾病的局部用制剂。
7.一种胸腺基质淋巴细胞生成素表达抑制剂,其特征在于,以抑制胸腺基质淋巴细胞生成素细胞因子的量包含龙眼肉、藁本及远志的组合生药提取物作为有效成分。
8.一种化妆品组合物,其特征在于,包含龙眼肉、藁本及远志的组合生药提取物作为抑制胸腺基质淋巴细胞生成素细胞因子或者治疗或改善皮肤炎症疾病的有效成分。
9.根据权利要求8所述的化妆品组合物,其特征在于,上述提取物使用选自水、包括甲醇、乙醇、丙醇、丁醇在内的C1-C4低级烷醇、丙酮、乙酸乙酯、三氯甲烷、己烷、丁二醇、丙二醇或甘油中的一种以上的溶剂提取。
10.根据权利要求8所述的化妆品组合物,其特征在于,上述皮肤炎症疾病为选自由因老化或特应性引起的瘙痒症、包括特应性皮炎、银屑病在内的慢性复发性皮炎、接触性皮炎、脂溢性皮炎、神经性皮炎、皮肤干燥症、红斑、炎性皮炎、银屑病或特应性疾病组成的组中的疾病。
11.根据权利要求8所述的化妆品组合物,其特征在于,上述组合物为选自润肤液、柔肤水、爽肤水、收敛剂、润肤乳、乳液、保湿乳液、滋养乳液、按摩霜、滋养霜、保湿霜、护手霜、粉底、精华、滋养精华、面膜、洁面泡沫、洁面乳液、卸妆面霜、润肤露、沐浴露、润发乳、美容液的形式。
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PCT/KR2021/002964 WO2021182865A1 (en) | 2020-03-13 | 2021-03-10 | A topical composition comprising an extract of combined herbs comprising longanae arillus for tlsp inhibition and the treatment or alleviation of skin inflammatory disease and the use thereof. |
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