CN101945886A - 采用含有脂质修饰半胱氨酸的肽的局部组合物和方法 - Google Patents
采用含有脂质修饰半胱氨酸的肽的局部组合物和方法 Download PDFInfo
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Abstract
本发明阐述用于局部施加至诸如皮肤或粘膜表面等角蛋白组织或基质的组合物。所述组合物可用于治疗皮肤病况或疾病。所述组合物包含至少一种多肽和/或脂化肽和载剂。
Description
相关申请案
本申请案主张优先于在2008年2月25日申请的美国临时专利申请案第61/067,161号,其全部揭示内容是以引用方式并入本文中。
背景技术
尽管期望健康皮肤,但持续暴露于体外要素中会引起皮肤损伤。所述损伤可因受伤、光损伤、萎缩、干性皮肤等引发。目前,市场上的许多洗剂和乳膏旨在帮助减轻皮肤损伤和/或相关不适。业内一直致力于发现可治疗和减轻皮肤损伤的新颖药剂。
发明内容
因此,本发明的第一方面涉及具有脂化半胱氨酸残基的分离肽或肽混合物,其中脂质是聚类异戊二烯,并且涉及包含所述肽或其混合物和适合局部投与的载剂的组合物。术语“局部投与”意欲包括直接或间接施加至角蛋白基质(例如皮肤、毛发或头皮)或施加至粘膜(例如经口投与)。
本发明第二方面涉及促进皮肤健康或减轻因暴露于各要素中所致皮肤损伤的方法,其包含直接或间接向皮肤施加包含具有脂化半胱氨酸残基的肽或肽混合物(其中脂质是聚类异戊二烯)和适合局部投与的载剂的组合物。
本发明第三方面涉及制备用于促进皮肤健康或减轻因暴露于各要素中所致皮肤损伤的局部组合物的方法,其包含调配包含具有脂化半胱氨酸残基的肽或肽混合物(其中脂质是聚类异戊二烯)和适合局部投与的载剂的组合物。
在本发明这些方面的某些实施例中,天然存在的肽或肽混合物的序列含有半胱氨酸,而在其它实施例中,将脂化半胱氨酸引入肽中。
在某些实施例中,肽长度为约2至200个氨基酸,而在其它实施例中,肽混合物的平均大小为约2至约200个氨基酸。
在某些实施例中,脂化半胱氨酸残基存于C-末端。在某些实施例中,脂化半胱氨酸残基是5个C-末端氨基酸残基中的至少一者。在某些实施例中,肽含有不止一个脂化半胱氨酸残基,其中不止一个半胱氨酸残基中的两者邻接或不邻接,并且其中肽的C-末端可为或可不为脂化半胱氨酸残基。在某些实施例中,肽含有非C-末端脂化半胱氨酸残基。在某些实施例中,肽混合物可在脂化肽中的不同氨基酸位置含有半胱氨酸残基。
在某些实施例中,存于半胱氨酸上的脂基是牻牛儿基、法呢基、植基、或牻牛儿基牻牛儿基,因此脂化残基分别称作牻牛儿基半胱氨酸(GC)、法呢基半胱氨酸(FC)、植基半胱氨酸(PC)、和牻牛儿基牻牛儿基半胱氨酸(GGC)。在某些实施例中,肽含有不止一个脂化半胱氨酸残基,其中每个半胱氨酸上的脂基相同或不同。
在某些实施例中,肽以占组合物约0.01%w/v至约10%w/v的量存在。
在某些实施例中,载剂是水基载剂并且组合物呈诸如凝胶或洗剂等乳液形式。在某些实施例中,载剂是非水基载剂(其中除了组合物的市售要素中存在的水以外,组合物另外不含外加水),其中组合物呈软膏剂或糊剂形式。在某些实施例中,组合物适合经口使用。
在某些实施例中,将纯净脂化肽与诸如水解丝蛋白等额外载剂掺合以获得均匀地经脂质修饰的肽。在某些实施例中,将均匀地经脂质修饰的肽调配为护发素。在某些实施例中,护发素含有碱。在某些实施例中,护发素含有水。在某些实施例中,护发素含有鲸蜡醇。
具体实施方式
局部组合物可含有含脂化半胱氨酸残基的肽或肽混合物和载剂。因此,本发明的肽一般具有至少一个脂化半胱氨酸残基,并且在本文中也称作脂化肽。本发明组合物可另外含有非脂化肽。
本发明的肽可含有多达200个或更多氨基酸残基,但在某些实施例中,肽长度较短。
本发明的肽可在肽的C-末端含有脂化半胱氨酸残基。在某些实施例中,半胱氨酸残基位于肽C-末端约5个残基以内。
在某些实施例中,肽含有不止一个半胱氨酸残基。在这些残基中,一个残基、某些残基或所有残基可经脂化。在所述情形下,当单一肽中存在两个或更多个半胱氨酸残基时,半胱氨酸残基可彼此相邻或一或多个氨基酸残基可分隔各半胱氨酸残基。
脂基可藉助诸如硫醚键等化学键结合至半胱氨酸残基。
本发明脂化肽可在(例如)N-末端区域含有其它化学修饰。可对肽中的N-末端或其它自由胺进行化学修饰,以增强脂化肽的溶解性、稳定性或渗透性。可用于增强脂化肽的溶解性、稳定性或渗透性的化学修饰技术为业内所熟知。例如,技术可包括乙酰化和/或BOC保护。可在肽中N-末端第一氨基酸的游离氨基处或附近进行修饰以改变肽的化学特性。可进行的其它修饰为所属领域技术人员所熟知。
可用于本发明目的的脂质是聚类异戊二烯,包括(例如)牻牛儿基、法呢基、牻牛儿基牻牛儿基和植基。可使用不止一种脂质来修饰肽中的半胱氨酸残基。例如,若肽含有两个或更多个半胱氨酸残基,则一个半胱氨酸残基可经植基脂化,同时另一半胱氨酸残基可经法呢基脂化。
在某些实施例中,含有肽之本发明组合物可有效用于局部应用中以促进上皮的健康或治疗上皮相关病症。这些本发明组合物在局部施加时不表现全身性效应。所述本发明组合物的安抚、增湿和去污特性尤其可用于治疗美容病况和/或一般用于促进皮肤的健康。本发明组合物可有效用于美容、药用化妆和一般的皮肤护理组合物以及医药组合物。
下文阐述本文所用某些术语的定义,其中许多为所属领域技术人员所理解。
说明书和权利要求书中通篇使用的词组“上皮”或“上皮的”或“上皮组织”意欲包括皮肤和粘膜。因此,本发明提供可用于治疗皮肤或粘膜病况的组合物,所述粘膜是(例如,但不限于)鼻粘膜、口腔粘膜、眼粘膜、耳粘膜、阴道粘膜和直肠粘膜。
在某些实施例中,本发明组合物包括医药组合物。本文所用“医药组合物”是指用于预防靶病况或疾病、降低其强度、治愈或以其它方式治疗靶病况或疾病的组合物。
在本文中术语“多肽”与“肽”可互换使用。在某些实施例中,“多肽”和“肽”可指具有2个或更多个(例如3、4、5、6、7、8、9、10、15、20、25、30、35、40、45、50、55、60、65、70、75、80、85、90、95、100、200、300、400、500或更多个)氨基酸的任何序列。在某些实施例中,氨基酸数量少于1000、500、400、300、200、100个左右。在某些实施例中,术语“二肽”是指具有2个氨基酸的“多肽”。
在某些实施例中,本发明组合物包括本文所定义的美容组合物。
1.实例性多肽的说明
本发明的肽可自任何已知适合局部施加至角蛋白组织的蛋白质、肽或肽混合物来生成。业内已知并且当前用于局部应用产品的蛋白质包括大豆蛋白、角蛋白、乳清、酪蛋白、丝蛋白、小麦蛋白、水稻蛋白、燕麦蛋白、和用于食品或美容产品的其它天然来源的蛋白质。
此外,本发明涵盖可使用细胞信号传导过程内的天然酶来生成本发明的肽。例如,可局部施加含有CaaX基序或脂化半胱氨酸的肽,并且随后通过内源性酶进行处理,从而在CaaX情形下加成脂质,并且在任一情形下移除半胱氨酸C-末端的氨基酸。
表I绘示113种以脂化形式存于人体内的人类蛋白的基因库(GENBANK)标识符和C-末端氨基酸序列。表I中包括源自G蛋白的序列。C-末端的四氨基酸序列构成CaaX基序。此基序中半胱氨酸残基后的三个氨基酸残基可在C-末端半胱氨酸残基脂化后移除。因此这些经修饰肽是本发明脂化肽的代表性实例。
表I-实例性多肽
在自表I选择肽(或多肽)或肽(或多肽)的组合用于本发明时,所属领域技术人员可注意到上游氨基酸残基之保守性。尽管C-末端半胱氨酸残基的上游(或相对于其的N-末端)氨基酸序列几乎不具有(若存在)同源性或序列相似性,但紧邻C-末端半胱氨酸残基上游的氨基酸残基仍具有一定保守性。表II展示紧邻CaaX基序中半胱氨酸残基上游的氨基酸残基的保守性。人们相信,氨基酸残基的保守性越高,用于细胞信号传导结合位点的特定肽的活性越大。例如,丝氨酸似乎是CaaX基序中半胱氨酸残基上游最普遍的氨基酸残基,并且因此紧邻C-末端区域中半胱氨酸残基上游具有丝氨酸残基的肽可在细胞调节和信号传导蛋白方面具有较强活性。不欲受限于任何特定理论或作用机制,本发明的肽可用于模拟或匹配多个序列(例如表I中的序列),并且因此活性强于单氨基酸模拟物,例如N-乙酰基-S-法呢基-半胱氨酸(AFC)。
表II
Q | 5 | 0.04424779 | 4.424779 |
F | 5 | 0.04424779 | 4.424779 |
T | 4 | 0.03539823 | 3.539823 |
N | 4 | 0.03539823 | 3.539823 |
H | 4 | 0.03539823 | 3.539823 |
I | 4 | 0.03539823 | 3.539823 |
E | 3 | 0.02654867 | 2.654867 |
Y | 3 | 0.02654867 | 2.654867 |
D | 2 | 0.01769912 | 1.769912 |
M | 1 | 0.00884956 | 0.884956 |
W | 1 | 0.00884956 | 0.884956 |
总计 | 113 |
可使用业内已知方法来制备具有期望长度的肽以达成本发明的目的。例如,可部分水解天然蛋白以获得具有约2-200个或更多个氨基酸残基的肽的混合物。
并非所有天然肽都含有半胱氨酸残基。然而,所述肽可经修饰以在肽序列中任一期望位置包括脂化半胱氨酸,例如在C-末端5个残基内通过引入半胱氨酸残基且随后加成脂基、或通过将脂化半胱氨酸残基直接引入肽序列中来修饰。例如,可使法呢基半胱氨酸(FC)在N或C-末端与肽偶合。参见实例1。
除源自天然蛋白来源(例如大豆蛋白、角蛋白、乳清、酪蛋白、蛋白质和诸如此类)的肽以外,可使用合成肽作为起始材料来合成可用于本发明目的的肽。与源自天然蛋白的肽类似,合成肽可经修饰以包括半胱氨酸残基。
在本发明的肽含有两个或更多个半胱氨酸残基并且期望之后对半胱氨酸残基进行脂化时,应理解,通过使用习用实验室方案,所有存于肽中的半胱氨酸残基都可经脂质修饰,即经单一脂质脂化。
本发明脂化肽可通过化学修饰或富集经脂质修饰肽来生成。
可使用化学修饰来提高经脂质修饰半胱氨酸残基在期望肽内的含量。在一实施例中,脂基和含有半胱氨酸的肽分开存在。可对肽进行化学修饰并且由此在半胱氨酸残基处进行脂化。例如,可在适宜条件下使游离半胱氨酸残基或含有半胱氨酸残基的肽与诸如溴化法呢基酯等活化脂质反应,从而使游离半胱氨酸残基发生脂化。参见实例2。
在另一实施例中,可脂化半胱氨酸残基并且随后可使用脂化半胱氨酸残基来修饰肽,从而使得脂化半胱氨酸残基位于C-末端。可通过用(例如)Fmoc或BOC保护天然衍生肽中的氨基并且随后用活化基团活化肽的C-末端并使诸如法呢基-半胱氨酸等经脂质修饰半胱氨酸与肽在一起反应来使所述半胱氨酸与肽的C-末端偶合。也可使经脂质修饰半胱氨酸与固相载体偶合以促进所述方法。参见实例3。
在一种此类方法中,通过以下方式来使诸如法呢基等脂质部分附接至半胱氨酸残基:使半胱氨酸的硫氢基与溴化法呢基酯在碱性条件下反应以附接法呢基。在另一方法中,通过使用脂质硫醇代替含有溴代丙氨酸的肽中的溴来制备脂化肽。例如,可根据美国专利第3,429,970号或吉尔伯特(Gilbert)等人,美国化学学会杂志(J.Am Chem.Soc.),114:3966-3913(1992)中所述的方法来合成法呢基硫醇。
已知本发明脂化肽也可能在自然界中存在。在所述情况下,期望富集脂化肽,在此情形下自其自然环境对其进行提取或分离。此通常涉及获取脂化肽的自然来源(即水解蛋白),和使用色谱法或化学分离来部分纯化混合物,并由此提高含有经脂质修饰半胱氨酸残基的肽在产物中的百分比。因此,经脂质修饰肽存于产物中的百分比高于初始蛋白质来源。
或者,也可富集已经化学修饰并由此脂化的本发明的肽。就这一点来说,可获得较高百分比的脂化肽或经脂质修饰半胱氨酸残基。
富集脂化肽或脂化半胱氨酸残基的方法为业内所熟知。例如,富集可包括使用特异性识别脂化肽的抗体或适体。参见吉尔伯特等人,生物有机和医学化学(Bioorganic and Medicinal Chemistry),5(6):1115-22(1997)。
此外,也可使用选择性基于脂质疏水性的管柱色谱、固相或液体提取来富集脂质修饰。在此情形下,可使用对脂质或经脂质修饰半胱氨酸残基具有一般或特异性亲和性的树脂,即C4树脂、与疏水染料分子偶合的树脂、与固体载体偶合的血清白蛋白、或具有定制官能团或结合位点的树脂。其它标准固相合成方法阐述于以下文献中:鲁姆柏瑞(Lumbierres)等人,11(24):7405-15(2005);帕卡穆萨(Pachamutha)等人,有机化学杂志(J.Org.Chem.),70(9):3720-3(2005);和多阿特(Douat)等人,肽科学杂志(J.Pept.Sci.),8(11):601-14(2002);和道伦斯(Dolence)等人,组合化学杂志(J.Comb.Chem.),2(5):522-36(2000),所有这些文献都是以引用方式并入本文中。参见实例3。
2.天然信号传导过程的调节
本发明的肽可为异戊烯半胱氨酸羧甲基转移酶(ICMT)的底物。ICMT是编码三种转译后修饰某些蛋白中的异戊烯化C-末端半胱氨酸残基并使所述蛋白靶向细胞膜的酶中的一者的基因。此酶位于内质网中。
与AFC类似,并且同样不欲受限于特定作用理论,本发明脂化肽可与细胞靶竞争ICMT的修饰。此外,本发明脂化肽也可与G蛋白或其它经法呢基、牻牛儿基牻牛儿基或棕榈酰基修饰的肽竞争相互作用位点。
例如,酵母a-因子是法呢基化肽,并且以与G蛋白相同的方式由相同酶甲基化。参见哈利塞纳(Hrycyna)等人,欧洲分子生物学协会杂志(EMBO J.),10(7):1699-709(1991)。可通过哺乳动物ICMT对合成法呢基化或牻牛儿基牻牛儿基化肽进行甲基化。参见斯蒂芬森(Stephenson)和克拉克J.(Clarke J.),生物与生物化学(Biol Chem.),265(27):16248-54(1990);吉纳尔(Giner)和兰多(Rando),生物化学(Biochemistry),33(50):15116-23(1994),其都是以引用方式并入本文中。
此外,经修饰肽亚组可能对其细胞靶具有较强亲和性,并且由此在生化和细胞分析中可具有优于AFC的潜能,并且由此在本发明精神中可用。不欲受限于任何特定理论,申请人相信情况确实如此,因为本发明脂化肽具有除AFC中的经修饰半胱氨酸残基以外的氨基酸。
3.局部制剂
本发明的肽可与适合局部投与的载剂一起使用并且因此被认为是美容、医药或皮肤病学上可接受的。载剂对诸如皮肤等角蛋白组织无毒性并且一般为惰性,但载剂也可提供益处。
局部制剂可均匀地经脂质修饰。换句话说,局部制剂内所包括的所有脂化肽都可经单一脂质修饰。另一方面,并非组合物中的每个单一肽都必须脂化。例如,在部分水解的蛋白质提取物中,并非所有肽都会含有半胱氨酸残基。因此,若用溴化法呢基酯处理,则仅含有半胱氨酸残基的部分会发生脂化(参见实例2)。因此,术语“均匀地经脂质修饰”也指局部制剂内的所有脂化肽都具有明确界定并且一致的分布。例如,局部制剂中所含的脂化肽可经两种或更多种脂质修饰并且具有明确界定并且一致的分布以使局部制剂均匀地经脂质修饰。
术语“局部的”是指在施加点或紧邻施加点下方投与本发明组合物。
局部施加在本文中是指施加至包括角蛋白组织在内的一或多个表面上,即“局部施用”。局部施加或“局部施用”可涉及直接施加至期望基质的区域中。若局部制剂和/或组合物为液体,则其可通过倾注、滴加、或喷雾来施用;若为软膏剂、洗剂、乳膏、凝胶或类似物则可通过擦抹来施用;若为粉剂则可通过撒施来施用;若为液体或气溶胶组合物则可通过喷雾来施用;或可通过任何其它适宜方式来施用。
在另一实施例中,局部制剂和/或组合物。本文所用“美容组合物”是指意欲擦抹、倾注、喷洒、或喷雾于基质或其任一部分上、引入基质或其任一部分中、或以其它方式施加至基质或其任一部分以清洁、美化、提高吸引力、或改变外观的组合物,或意欲用作任一物件中的一种组份的所述物件。
脂化肽中以一般占局部组合物0.01%至约10%w/v范围内的浓度存于局部组合物。在某些实施例中,脂化肽的量可超过10%w/v。然而,更通常来说,脂化肽以介于0.1%与5%w/v之间的浓度存于局部组合物中。所述量被视为可“有效”用于本文所述使用所述组合物的目的。
载剂要素的选择取决于所设想调配物的类型。可将局部制剂局部施加至皮肤、毛发、头皮或粘膜且其可呈任何形式,包括溶液、油、乳膏(例如增湿剂)、软膏剂、乳液(例如洗发剂)、悬浮液、凝胶、洗剂、乳状物、清洁剂、喷雾剂、透皮贴剂和诸如此类。参见实例4。
在另一实施例中,聚异戊二烯蛋白抑制剂化合物、载剂与任选地其它活性成份组成组合物。
可通过在全部诸如水或有机溶剂等溶剂载剂(例如低碳醇(例如低碳烷醇,例如乙醇和异丙醇)和丙酮)中均匀混合溶质或经溶解物质(例如脂化肽和任选地一或多种活性成份)来制备溶液。基本乳液含有至少三种组份,两种不可混溶液体载剂和乳化剂以及脂化肽。大多数乳液将水性相纳入非水性相中(或反之亦然)。然而,可制备基本上非水性的乳液,例如非水性不可混溶系统甘油和橄榄油的阴离子型和阳离子型表面活性剂。凝胶与乳液的不同之处一方面在于其不含乳化剂。
在另一实施例中,脂化肽可与凝胶悬浮液(半固体载剂)或固体载剂混合以形成糊剂、粉剂、软膏剂、乳膏、洗剂、水凝胶或诸如此类。
例如,可制备呈凝胶悬浮液形式的软膏剂。这些软膏剂是意欲外部施加至上皮的半固体制剂。一般来说,软膏剂基质可分类为烃基质(油质),其可使用白凡士林作为基质;吸附性基质(无水),其可使用亲水矿脂或无水羊毛脂;乳液基质(油包水型);乳液基质(水包油型);和水溶性基质,其通常使用聚乙二醇作为软膏剂基质。
其它使用脂化肽和载剂的局部制剂可使用业内已知的技术容易地制备,例如以下文献中所述:雷明顿制药科学(Remington′s Pharmaceutical Sciences),第18或19版,麦克出版公司(Mack Publishing Company),伊斯顿(Easton),宾夕法尼亚州(Pa)出版。
因此,载剂的非限制性代表性实例包括增湿剂或湿润剂、pH调节剂、除臭剂、香气剂、护发素、螯合剂、防腐剂、乳化剂或表面活性剂、增稠剂、增溶剂、渗透促进剂、抗刺激剂及着色剂。
本文所用“增湿剂”是使皮肤增加或恢复湿润的物质。可用于本发明中的增湿剂或湿润剂的代表性实例包括(但不限于)胍、羟乙酸和羟乙酸盐(例如铵盐和烷基季铵盐)、呈多种形式中的任一种的真芦荟(aloe vera)(例如真芦荟凝胶)、尿囊素、尿唑、多元醇(例如山梨醇、甘油、己三醇、丙二醇、丁二醇、己二醇和诸如此类)、聚乙二醇、糖和淀粉、糖和淀粉衍生物(例如烷氧基化葡萄糖)、透明质酸、乳酰胺单乙醇胺、乙酰胺单乙醇胺和其任一组合。
如业内所广泛认可,由于皮肤的pH为5.5,因此为避免刺激,皮肤用局部制剂的pH值优选地应介于4.0与7.0之间,优选地介于5.0与6.0之间,最优选地为约5.5或大致为5.5。因此,通常添加pH调节组合物以使组合物的pH达到期望值。因此,局部制剂优选地经调配以使pH值介于约4.0与约7.0之间的范围内,更优选地介于约5.0与约6.0之间。
适宜pH调节剂包括(例如,但不限于)一或多种肥酸、甘氨酸、柠檬酸、氢氧化钙、硅酸铝镁、缓冲剂或其任一组合。
本文所用“除臭剂”是指用于抑制或遮蔽流汗或其它体味的物质。可用除臭剂的代表性实例包括(但不限于)季铵化合物(例如溴化十六烷基三甲铵、氯化十六烷基吡啶、苄索氯铵(benzethonium chloride)、二异丁基苯氧基乙氧基乙基二甲基苯甲基氯化铵、N-十二烷基肌氨酸钠、N-棕榈酰肌氨酸钠、月桂酰肌氨酸、N-肉豆蔻酰甘氨酸、N-十二烷基肌氨酸钾、十八烷基三甲基氯化銨、乳酸氯氢化铝钠、三-十六烷基甲基氯化铵)、2,4,4’-三氯-2’-羟基二苯醚、二氨基烷基酰胺(例如L-赖氨酸十六烷基酰胺)、柠檬酸、水杨酸、和吡罗克酮(piroctose)的重金属盐(尤其锌盐)和其酸、巯氧吡啶(pyrithione)的重金属盐(尤其巯氧吡啶锌)和酚磺酸锌。其它除臭剂包括(但不限于)气味吸收材料,例如碳酸盐和碳酸氢盐,例如碱金属碳酸盐和碳酸氢盐、铵和四烷基铵碳酸盐和碳酸氢盐,尤其钠盐和钾盐,或上述任一组合。本发明组合物中可以溶解或颗粒形式纳入止汗剂,且其包括(例如)铝或锆收敛剂盐或络合物。
本文所用“香气剂”是指具有香气的物质。适宜香气剂包括(但不限于)桉叶油、合成樟脑、薄荷油、丁香油、薰衣草、甘菊和诸如此类。
可用适宜护发素包括(例如)一或多种胶原、阳离子型表面活性剂、经修饰硅酮、蛋白质、角蛋白、二甲硅油多元醇、季铵化合物、卤代季铵化合物、烷氧基化羧酸、烷氧基化醇、烷氧基化酰胺、山梨糖醇酐衍生物、酯、聚醚、甘油酯、或其任一组合。
任选地可将螯合剂添加至局部制剂中以增强防腐性或防腐系统。优选螯合剂是温和试剂,例如乙二胺四乙酸(EDTA)、EDTA衍生物、或其任一组合。
适宜使用的防腐剂可包括(但不限于)一或多种烷醇、EDTA二钠(乙二胺四乙酸盐)、EDTA盐、EDTA脂肪酸共轭物、异噻唑啉酮、对羟基苯甲酸酯类(例如对羟基苯甲酸甲酯和对羟基苯甲酸丙酯)、丙二醇、山梨酸酯、脲衍生物(例如重氮烷基咪唑脲)、或其任一组合。
本文所用乳化剂(也称作表面活性剂)促进乳液的形成和稳定。适宜乳化剂可为天然材料、精细固体、或合成材料。天然乳化剂可得自动物或植物来源。来自动物来源的乳化剂可包括明胶、蛋黄、酪蛋白、羊毛脂、或胆固醇。来自植物来源的乳化剂可包括阿拉伯胶、黄蓍胶、鹿角菜胶、或果胶。明确来自植物来源的纤维素衍生物包括甲基纤维素和羧甲基纤维素以提高粘度。精细乳化剂可包括皂粘土、氢氧化镁、氢氧化铝或三硅酸镁。合成试剂是阴离子型、阳离子型、两性离子型或非离子型。用于局部组合物中的乳化剂的实例包括十二烷基硫酸钠、苯扎氯铵(benzalkonium chloride)、聚乙二醇400单硬脂酸酯、和其组合。
本文所用“增稠剂”(也称作胶凝剂)是指使局部制剂在稠度方面更稠密或更粘稠的试剂。增稠剂是水基或油基试剂。可用于本发明背景中的适宜增稠剂包括(例如)非离子型水溶性聚合物,例如羟乙基纤维素(以商标NatrosolTM 250或350购得);阳离子型水溶性聚合物,例如Polyquat 37(以商标SynthalenTM CN购得);脂肪醇、脂肪酸、阴离子型聚合物、和其碱金属盐以及其混合物。
本文所用“增溶剂”是使溶质可溶解的物质。可用于本文背景中的增溶剂的代表性实例包括(但不限于)络合物形成性增溶剂,例如柠檬酸、乙二胺四乙酸盐、偏磷酸钠、琥珀酸、尿素、环糊精、聚乙烯吡咯烷酮、邻苯甲酸二乙铵;和胶束形成增溶剂,例如TWEENTM和司盘(spans),例如TWEEN 80TM。可用于局部制剂的其它增溶剂可为(例如)聚氧乙烯山梨糖醇酐脂肪酸酯、聚氧乙烯N-烷基醚、N-氧化N-烷基胺、泊洛沙姆(poloxamer)、有机溶剂(例如丙酮)、磷脂和环糊精。
“渗透促进剂”是已知可加速物质穿过皮肤递送的试剂。适宜渗透促进剂可包括(但不限于)二甲亚砜(DMSO)、二甲基甲酰胺(DMF)、尿囊素、尿唑、N,N-二甲基乙酰胺(DMA)、癸基甲基亚砜(C10MSO)、聚乙二醇单月桂酸酯(PEGML)、丙二醇(PG)、丙二醇单月桂酸酯(PGML)、甘油单月桂酸酯(GML)、卵磷脂、1-取代氮杂环庚-2-酮、尤其1-正十二烷基环氮杂环庚-2-酮(可以商标AzoneTM自维特白研究集团(Whitby Research Incorporated),里士满,Va.购得)、醇类和诸如此类。渗透促进剂也可为植物油。所述油可包括(例如)红花油、棉籽油和玉米油。
其它增稠剂、渗透促进剂和其它载剂材料或佐剂一般可参见雷明顿制药科学,第18或19版,由宾夕法尼亚州伊斯顿的麦克出版公司出版,其是以引用方式并入本文中。
本文所用“抗刺激剂”是可预防或减少身体部分中的酸痛、粗糙度、或炎症的试剂。可用适宜抗刺激剂包括(例如,但不限于)类固醇和非类固醇消炎剂或其它材料,例如真芦荟、甘菊、α-没药醇、可乐果(cola nitida)提取物、绿茶提取物、茶树油、甘草提取物、尿囊素、咖啡因或其它黄嘌呤、甘草酸和其衍生物。
可将当前已知的抗刺激剂分为水溶性抗刺激剂和水不溶性抗刺激剂。所述组合物的代表性实例阐述于(例如)美国专利第5,482,710号中,其是以引用方式并入本文中。
着色剂可包括颜料或染料或其组合。优选颜料可包括(但不限于)氧化铁、和氧化钛。适宜染料包括FD&C批准的着色剂、D&C批准的着色剂、和那些批准在欧洲和日本使用的染料。参见玛密翁D.M.(Marmion,D.M.),美国用于食品、药物、化妆品、和医疗设备的着色剂手册(Handbook of US Colorants for Food,Drugs,Cosmetics,and Medical Devices),第3版,1991,其是以引用方式并入本文中。
在某些实施例中,本发明组合物中包括医药上可接受的载剂。本文所用“医药上可接受的载剂”是任何传统上可用于局部投与药物的基本无毒载剂,其中在直接施加至皮肤或粘膜表面时肽可保持稳定和生物可利用性。
在某些实施例中,本发明组合物包括美容上可接受的载剂。本文所用词组“美容上可接受的载剂”是指传统上可用于局部投与化妆品的基本无毒载剂,其中本发明多肽可保持稳定和生物可利用性。所属领域技术人员应理解,美容上可接受的载剂和医药上可接受的载剂的性质相似但通常并不完全相同。
本发明其它适宜载剂可包括医药上可接受的载剂。例如,水、凡士林(VASELINE)、矿物油、植物油、动物油、有机和无机蜡(例如微晶、石蜡和地蜡)、天然聚合物(例如苍耳烷、明胶、纤维素、胶原、淀粉、或阿拉伯胶)、醇、多元醇、CTFA国际化妆品成份字典和手册(CTFA International Cosmetic Ingredient Dictionary and Handbook)(第8版,维尼格(Wenninger)和坎特伯雷(Canterbery)编辑,美容品、化妆品和香气剂联合公司(The Cosmetic,Toiletry,and Fragrance Association,Inc.),华盛顿D.C.,2000,其是以引用方式并入本文中)中所述的载剂、和诸如此类。也包括本文所述的其它载剂。
本发明组合物中的载剂可提供持续释放或延迟释放效应。载剂可为能持续或延迟释放脂化肽的任一材料以提供更有效的投与,从而降低脂化肽投用的频率和/或减少其剂量,使其易于处理,并且延长或延迟对上皮相关病况的作用。所述载剂的非限制性实例包括天然和合成聚合物的脂质体、微型海绵、微球体、或微胶囊和诸如此类。可促进脂化肽在皮肤各层内的局部递送的脂质体可自多种磷脂(例如胆固醇)、硬脂酰胺或磷脂酰胆碱来形成。
在某些实施例中,本发明组合物中的载剂含有成膜聚合物。具有成膜聚合物的局部载剂的实例包括防晒霜(美国专利第5,653,965号)、局部消毒剂(美国专利第7,323,163号)、和屏障组合物(美国专利第6,210,688号)。其它实例包括含有市售成膜聚合物的载剂,所述市售成膜聚合物是(例如)脱氢黄原胶、Eastman AQ 38S、Dermacryl AQFDermacryl 79Dermacryl LTDermacryl CAquamere、AvalureTM、和诸如此类。此一成膜聚合物可赋予防水特征或延迟释放特征。
在某些实施例中,可将局部制剂纳入适合投与口腔粘膜的载剂中,例如漱口剂、冲洗剂、口腔喷雾剂、悬浮液、牙科用凝胶、和诸如此类。业内已知的典型口腔用载剂可用于本发明中。口腔用媒剂的pH值一般为约4至约7,并且优选地为约5至约6.5。口腔局部制剂可另外含有通常用于诸如以下等产物中的习用添加剂:提供氟的化合物和甜味剂,前提是所述添加剂不干扰本发明组合物的治疗性或美容性有益特性。
提供氟的化合物可具有完全水溶性或微溶性,并且特征在于其能在水中释放氟离子或含氟离子并且其不能与组合物中的其它组份反应。典型的提供氟的化合物是无机氟化物盐,例如水溶性碱金属盐、碱土金属盐和重金属盐,例如氟化钠、氟化钾、氟化铵、氟化亚铜、氟化锌、氟化锡、氟化亚锡、氟化钡、氟硅酸钠、氟硅酸铵、氟锆酸钠、单氟磷酸钠、单氟磷酸铝和二氟磷酸铝、和氟化磷酸钙钠。优选者为碱金属氟化物、氟化锡和单氟磷酸盐,例如氟化钠和氟化亚锡、单氟磷酸钠和其混合物。
存在的提供氟的化合物的量可取决于所用提供氟的化合物的类型、氟化合物的溶解性、和最终口腔局部制剂的性质。所用提供氟的化合物的量必须为无毒量。一般来说,所用提供氟的化合物以口腔局部制剂的重量计可以最多约1%、优选地约0.001%至约0.1%、并且最优选地约0.001%至约0.05%的量存在。
在使用甜味剂(增甜剂)时,可采用业内熟知的增甜剂,包括天然和人工增甜剂。所用甜味剂可选自众多种材料,包括(但不限于)水溶性甜味剂、水溶性人工甜味剂、衍生自天然水溶性甜味剂的水溶性甜味剂、基于二肽的甜味剂、和基于蛋白质的甜味剂,包括其混合物。
局部组合物可另外包括一或多种其它相容性活性成份,此旨在向局部制剂提供除脂化肽所提供的效应以外的另一种医药、药用化妆或美容效应。本文所用“相容性”意指此一组合物中的各组份能以在普通使用条件下不发生可显著降低局部制剂效力的相互反应的方式彼此组合。
本文所用词组“另一活性成份”是指除脂化肽以外的药剂,其能发挥医药活性、皮肤病学活性或诸如美容效应等任何其它有益活性。应理解,“其它有益活性”可为只有使用本文所述局部制剂的个体才能体会出有益的活性。
因此,含有脂化肽并且另外包括一或多种其它活性成份的局部制剂除了作为治疗上皮相关病况的药物或仅作为化妆品的用途以外,可具有其它有效用途。
其它活性成份可包括(但不限于)以下试剂中一或多者的任一组合:保护剂、软化剂、收敛剂、刺激剂、角质分离剂、防晒剂、仿晒剂、抗生素试剂、抗真菌剂、抗病毒剂、抗原虫剂、祛痘剂、麻醉剂、类固醇消炎剂、非类固醇消炎剂、止痒剂、抗氧化剂、化学治疗剂、抗组胺剂、维生素、激素、去头屑剂、抗皱剂、抗皮肤萎缩剂、硬化剂、清洁剂、腐蚀剂和色素淡化剂。
“保护剂”是任何可将皮肤或其它膜的暴露表面与有害或扰人刺激物隔离的试剂。本文所述保护剂可呈以下形式:撒施粉剂、吸附剂、机械保护剂、和硬膏剂。撒施粉剂是用于覆盖和保护上皮表面、溃疡和伤口的相对惰性和不溶性材料。通常,这些物质是精细粉剂,其可吸收水分并且可用作干燥剂。对皮肤水分的吸收可减少摩擦并且也抑制某些细菌生长。用作保护性吸附剂的某些材料包括皂粘土、不溶性铋盐、硼酸、碳酸钙(沉淀的)、纤维素、玉米淀粉、硬脂酸镁、滑石粉、二氧化钛、氧化锌、和硬脂酸锌。
也可将保护剂投与皮肤以形成粘附性连续膜,其可为柔性或半硬性,此取决于材料和调配物以及其施用方式。此材料可用于多种目的,包括遮挡外部环境、提供化学载体、和用作其它药物的媒剂。机械保护剂一般为火棉胶或硬膏剂。实例可包括氢氧化铝凝胶、火棉胶、二甲硅油、凡士林纱布、吸收性明胶膜、吸收性明胶海绵、锌明胶、高岭土、羊毛脂、无水羊毛脂、矿物油、矿物油乳液、轻质矿物油、橄榄油、花生油、矿脂、硅酮、水胶体和诸如此类。
可涵盖的一个保护剂实例是缓和剂。缓和剂是主要用于减轻刺激、尤其是对粘膜或擦伤组织的刺激的保护剂。其经常以可快速覆盖某区域并可加有药品的半流体粘性制剂形式施加至表面。多种化学物质具有缓和剂特性。这些物质可包括藻酸盐、胶浆剂、树胶、糊精、淀粉、某些糖、和聚合多羟基二醇。其它物质包括阿拉伯胶、琼脂、安息香、卡波姆(carbomer)、明胶、甘油、羟乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、丙二醇、藻酸钠、黄蓍胶、水凝胶和诸如此类。
“软化剂”一般是温和的脂质或油质材料,其可局部施用,尤其施加至皮肤。软化剂增加组织水分含量,由此使皮肤更软更柔韧。可通过以下方式来使皮肤中的水分含量增加:用闭塞性水不可混溶屏障来防止水分流失,用湿润剂来提高皮肤的锁水能力,或改变皮肤最外层的角质层的脱皮。可用软化剂包括羊毛脂、鲸蜡、矿物油、石蜡、矿脂、白软膏剂、白凡士林、黄软膏剂。也包括植物油、蜡、十六烷基醇、甘油、亲水矿脂、肉豆蔻酸异丙酯、肉豆蔻醇、和油醇。
“收敛剂”一般是局部施用的蛋白质沉淀剂,其细胞渗透性较低以致于其作用主要限于细胞表面和间质间隙。收敛剂的作用伴随有组织的收缩和起皱以及黄化现象。收敛剂以治疗性方式用于通过凝固血液来阻止出血、促进愈合、使皮肤坚韧或减少出汗。收敛剂的主要组份是铝盐、锌盐、锰盐、铁盐或铋盐。
“刺激剂”是可局部作用于皮肤以诱导(基于刺激剂浓度)充血、炎症、和干燥的材料。刺激剂包括(但不限于)醇、芳香氨醑、安息香酊、樟脑、辣椒、和煤焦油提取物。优选地,刺激剂是发赤剂。本文所用“发赤剂”是可诱导充血的试剂,其中充血意指身体一部分或器官中的血量增加。发赤剂所诱导的发红是由于流向受伤区域的循环增加所致,并且伴随有舒适、温暖、瘙痒和感觉过敏的感觉。
可用于本发明背景中的防晒剂的代表性实例可包括(但不限于)对氨基苯甲酸和其盐和其衍生物(乙基酯、异丁基酯、甘油酯;对二甲基氨基苯甲酸);邻氨基苯甲酸酯(即邻氨基-苯甲酸酯;甲酯、薄荷基酯、苯基酯、苄基酯、苯乙基酯、沉香基酯、松油基酯、和环己烯基酯);水杨酸酯(戊基酯、苯基酯、辛基酯、苄基酯、薄荷基酯、甘油酯、和二丙二醇酯);肉桂酸衍生物(薄荷基酯和苄基酯、α-苯基肉桂腈;丁基肉桂酰基丙酮酸酯);二羟基肉桂酸衍生物(伞形酮、甲基伞形酮、甲基乙酰伞形酮);三羟基-肉桂酸衍生物(七叶亭(esculetin)、甲基七叶亭、瑞香素(daphnetin)、和葡糖苷、七叶苷(esculin)和瑞香苷(daphnin));烃(二苯基丁二烯、均二苯乙烯);二苄基丙酮和苄基苯乙酮;萘酚磺酸盐(2-萘酚-3,6-二磺酸和2-萘酚-6,8-二磺酸的钠盐);二羟基萘甲酸和其盐;邻-和对羟基二苯基二磺酸酯;香豆素(coumarin)衍生物(7-羟基、7-甲基、3-苯基);二唑(2-乙酰基-3-溴吲唑、苯基苯并噁唑、甲基萘噁唑、各种芳基苯并噻唑);奎宁(quinine)盐(硫酸氢盐、硫酸盐、氯化物、油酸盐、和鞣酸盐);喹啉衍生物(8-羟基喹啉盐、2-苯基喹啉);羟基或甲氧基取代的二苯甲酮;尿酸和紫尿酸;鞣酸和其衍生物(例如六乙醚);(丁基卡必醇(carbotol))(6-丙基胡椒基)醚;氢醌;二苯甲酮(苯酚、舒利苯酮(sulisobenzone)、二羟苯宗(dioxybenzone)、苯酰间二苯酚、2,2’,4,4’-四羟基二苯甲酮、2,2’-二羟基-4,4’-二甲氧基二苯甲酮、奥他苯酮(octabenzone);4-异丙基二苯甲酰甲烷;丁基甲氧基二苯甲酰甲烷;氰双苯丙烯酸乙酯;氰双苯丙烯酸辛酯;[3-(4’-甲基苯亚甲基莰-2-酮)和4-异丙基-二苯甲酰甲烷;和其任一组合。
可用于本发明中的免晒仿晒剂的代表性实例可包括(但不限于)二羟基丙酮、甘油醛、吲哚和其衍生物。免晒仿晒剂可与防晒霜剂组合使用。
本文所用术语“抗生素试剂”意指能抑制细菌和其它微生物生长或对其进行破坏的化学物质群组中的任一者,其主要用于治疗传染病。抗生素试剂的实例包括(但不限于)青霉素G(Penicillin G)、甲氧西林(Methicillin)、萘夫西林(Nafcillin)、苯唑西林(Oxacillin)、氯唑西林(Cloxacillin)、双氯西林(Dicloxacillin)、氨苄西林(Ampicillin)、阿莫西林(Amoxicillin)、替卡西林(Ticarcillin)、羧苄西林(Carbenicillin)、美洛西林(Mezlocillin)、阿洛西林(Azlocillin)、哌拉西林(Piperacillin)、亚胺培南(Imipenem)、氨曲南(Aztreonam)、头孢噻吩(Cephalothin)、头孢克洛(Cefaclor)、头孢西丁(Cefoxitin)、头孢呋辛(Cefuroxime)、头孢尼西(Cefonicid)、头孢美唑(Cefmetazole)、头孢替坦(Cefotetan)、头孢丙烯(Cefprozil)、氯碳头孢(Loracarbef)、头孢他美(Cefetamet)、头孢哌酮(Cefoperazone)、头孢噻肟(Cefotaxime)、头孢唑肟(Ceftizoxime)、头孢曲松(Ceftriaxone)、头孢他啶(Ceftazidime)、头孢吡肟(Cefepime)、头孢克肟(Cefixime)、头孢泊肟(Cefpodoxime)、头孢磺啶(Cefsulodin)、氯罗沙星(Fleroxacin)、萘啶酸(Nalidixic acid)、诺氟沙星(Norfloxacin)、环丙沙星(Ciprofloxacin)、氧氟沙星(Ofloxacin)、依诺沙星(Enoxacin)、洛美沙星(Lomefloxacin)、西诺沙星(Cinoxacin)、多西环素(Doxycycline)、米诺环素(Minocycline)、四环素(Tetracycline)、阿米卡星(Amikacin)、庆大霉素(Gentamicin)、卡那霉素(Kanamycin)、奈替米星(Netilmicin)、妥布霉素(Tobramycin)、链霉素(Streptomycin)、阿奇霉素(Azithromycin)、克拉霉素(Clarithromycin)、红霉素(Erythromycin)、依托红霉素(Erythromycin estolate)、琥乙红霉素(Erythromycin ethyl succinate)、葡庚糖酸红霉素(Erythromycin glucoheptonate)、乳糖酸红霉素(Erythromycin lactobionate)、硬酯酸红霉素(Erythromycin stearate)、万古霉素(Vancomycin)、替考拉宁(Teicoplanin)、氯霉素(Chloramphenicol)、克林霉素(Clindamycin)、甲氧苄啶(Trimethoprim)、磺胺甲噁唑(Sulfamethoxazole)、呋喃妥因(Nitrofurantoin)、利福平(Rifampin)、莫匹罗星(Mupirocin)、甲硝唑(Metronidazole)、头孢氨苄(Cephalexin)、罗红霉素(Roxithromycin)、辅-阿莫氟酸盐(Co-amoxiclavuanate)、哌拉西林与他唑巴坦(Tazobactam)的组合、和其各种盐、酸、碱和其它衍生物。抗细菌性抗生素试剂包括(但不限于)青霉素、头孢菌素(cephalosporin)、碳头孢烯(carbacephem)、头霉素(cephamycin)、碳青霉烯(carbapenem)、单酰胺菌素(monobactam)、氨基葡糖苷、糖肽、喹诺酮(quinolone)、四环素、大环内酯、和氟喹诺酮。
本文所用术语“抗真菌剂”意指能抑制真菌生长或破坏真菌的化学物质群组中的任一者。抗真菌剂包括(但不限于)两性霉素B(Amphotericin B)、克念霉素(Candicidin)、制皮菌素(Dermostatin)、非律平(Filipin)、制霉色基素(Fungichromin)、曲古霉素(Hachimycin)、哈霉素(Hamycin)、鲁斯霉素(Lucensomycin)、美帕曲星(Mepartricin)、那他霉素(Natamycin)、制霉菌素(Nystatin)、培西洛星(Pecilocin)、真菌霉素(Perimycin)、偶氮丝氨酸、灰黄霉素(Griseofulvin)、寡霉素(Oligomycin)、新霉素(Neomycin)、吡咯尼群(Pyrrolnitrin)、西卡宁(Siccanin)、杀结核菌素(Tubercidin)、绿毛菌素(Viridin)、布替萘芬(Butenafine)、萘替芬(Naftifine)、特比萘芬(Terbinafine)、联苯苄唑(Bifonazole)、布康唑(Butoconazole)、氯登妥因(Chlordantoin)、氯米达唑(Chlormidazole)、氯康唑(Cloconazole)、克霉唑(Clotrimazole)、益康唑(Econazole)、恩康唑(Enilconazole)、芬替康唑(Fenticonazole)、氟曲马唑(Flutrimazole)、异康唑(Isoconazole)、酮康唑(Ketoconazole)、拉诺康唑(Lanoconazole)、咪康唑(Miconazole)、奥莫康唑(Omoconazole)、奥昔康唑(Oxiconazole)、舍他康唑(Sertaconazole)、硫康唑(Sulconazole)、噻康唑(Tioconazole)、托西拉酯(Tolciclate)、托林达酯(Tolindate)、托萘酯(Tolnaftate)、氟康唑(Fluconazole)、伊曲康唑(Itraconazole)、沙康唑(Saperconazole)、特康唑(Terconazole)、吖啶琐辛(Acrisorcin)、阿莫罗芬(Amorolfine)、苯柳胺酯(Biphenamine)、溴柳氯苯胺(Bromosalicylchloranilide)、丁氯柳胺(Buclosamide)、丙酸钙、氯苯甘醚(Chlorphenesin)、环吡酮(Ciclopirox)、氯羟喹(Cloxyquin)、科帕腊芬内特(Coparaffinate)、双胺噻唑(Diamthazole)、依沙酰胺(Exalamide)、氟胞嘧啶、胺氯苯噻唑(Halethazole)、海克替啶(Hexetidine)、氯氟卡班(Loflucarban)、硝呋太尔(Nifuratel)、碘化钾、丙酸、巯氧吡啶、水杨苯胺、丙酸钠、舒苯汀(Sulbentine)、替诺尼唑(Tenonitrozole)、三醋汀(Triacetin)、苄硫噻二嗪乙酸(Ujothion)、十一烯酸、和丙酸锌。
本文所用术语“抗病毒剂”意指能抑制病毒复制或破坏病毒的化学物质群组中的任一者,其主要用于治疗病毒性疾病。抗病毒剂包括(但不限于)无环鸟苷(Acyclovir)、西多福韦(Cidofovir)、阿糖胞苷(Cytarabine)、双脱氧腺苷、去羟肌苷、依度尿苷(Edoxudine)、泛昔洛韦(Famciclovir)、氟尿苷(Floxuridine)、更昔洛韦(Ganciclovir)、碘苷(Idoxuridine)、肌酐普拉诺贝(Inosine Pranobex)、拉米夫定(Lamivudine)、MADU、喷昔洛韦(Penciclovir)、索立夫定(Sorivudine)、司他夫定(Stavudine)、曲氟尿苷(Trifluridine)、伐昔洛韦(Valacyclovir)、阿糖腺苷(Vidarabine)、扎西他滨(Zalcitabine)、齐多夫定(Zidovudine)、醋孟南(Acemannan)、乙酰基亮氨酸、金刚烷胺、粘病毒霉素(Amidinomycin)、地拉韦啶(Delavirdine)、膦甲酸(Foscamet)、茚地那韦(Indinavir)、干扰素-α、干扰素-β、干扰素-γ、乙氧丁酮醛(Kethoxal)、溶菌酶、美替沙腙(Methisazone)、吗啉胍(Moroxydine)、奈韦拉平(Nevirapine)、鬼臼毒素(Podophyllotoxin)、利巴韦林(Ribavirin)、金刚乙胺、利托那韦2(Ritonavir 2)、沙奎那韦(Saquinavir)、斯塔利霉素(Stailimycin)、维斯托隆(Statolon)、曲金刚胺(Tromantadine)、齐多夫定(AZT)和珍那佐酸(Xenazoic Acid)。
本文所用术语“抗原虫剂”意指能抑制原虫生长或破坏原虫的化学物质群组中的任一者,其主要用于治疗原虫性疾病。抗原虫剂的实例包括(但不限于)乙胺嘧啶(pyrimethamine)(DARAPRIM)、磺胺嘧啶(sulfadiazine)和甲酰四氢叶酸(Leucovorin)。
适宜祛痘剂可包括(但不限于)角质分离剂,例如水杨酸、硫酸、羟乙酸、丙酮酸、间苯二酚、和N-乙酰基半胱氨酸;和类视色素,例如视黄酸和其衍生物(例如顺式和反式视黄酸、视黄酸酯)和视黄醇。
“麻醉剂”是指可使感觉降低或损失的药剂。适宜麻醉药的非限制性实例可包括以下物质的医药上可接受的盐:利多卡因(lidocaine)、布比卡因(bupivacaine)、氯普鲁卡因(chlorprocaine)、地布卡因(dibucaine)、依替卡因(etidocaine)、甲哌卡因(mepivacaine)、丁卡因(tetracaine)、达克罗宁(dyclonine)、海克卡因(hexylcaine)、普鲁卡因(procaine)、可卡因(cocaine)、氯胺酮(ketamine)、普拉卡因(pramoxine)和酚。
本文所用术语“医药上可接受的盐”可包括(但不限于)可能存于本发明化合物中的酸性或碱性基团的盐。碱性化合物能与不同无机和有机酸形成众多种盐。所述无毒盐(即含有药理上可接受的阴离子的盐)可包括(但不限于)盐酸盐、氢溴酸盐、氢碘酸盐、硝酸盐、硫酸盐、硫酸氢盐、磷酸盐、酸式磷酸盐、异烟酸盐、乙酸盐、乳酸盐、水杨酸盐、柠檬酸盐、酸式柠檬酸盐、酒石酸盐、草酸盐、油酸盐、鞣酸盐、泛酸盐、酒石酸氢盐、抗坏血酸盐、琥珀酸盐、马来酸盐、龙胆酸盐、富马酸盐、葡萄糖酸盐、葡糖醛酸盐、糖二酸盐、甲酸盐、苯甲酸盐、谷氨酸盐、甲磺酸盐、乙磺酸盐、苯磺酸盐、对甲苯磺酸盐和双羟萘酸盐(即1,1’-亚甲基-双-(2-羟基-3-萘酸盐))。可包括氨基的本发明化合物也可与除上述酸以外的各种氨基酸形成医药上可接受的盐。酸性化合物能与各种药理上可接受的阳离子形成碱盐。所述盐的实例可包括碱金属盐或碱土金属盐,并且尤其包括钙盐、镁盐、钠盐、锂盐、锌盐、钾盐和铁盐。其它所述盐可包括医药上可接受的有机碱,例如氨、精氨酸、苯乙苄胺(benethamine)、苄星青霉素(benzathine)、地阿诺(deanol)、二乙醇胺、二乙胺、-2-二乙氨基乙醇、乙醇胺、乙二胺、赖氨酸、-2-羟乙基吗啉、哌嗪、-2-羟乙基吡咯烷、三乙醇胺、氨丁三醇(tromethamine)。其它所述盐可包括N-保护肽的盐。
本文所用“类固醇消炎剂”是指多种含有17碳4环系统的化合物中的任一种,并且包括固醇、各种激素(例如促蛋白合成类固醇)、和糖苷。类固醇消炎剂的代表性实例包括(但不限于)皮质类固醇,例如氢化可的松(hydrocortisone)、羟基曲安西龙(hydroxyltriamcinolone)、α-甲基地塞米松(α-methyl dexamethasone)、磷酸地塞米松、丙酸倍氯米松(beclomethasone dipropionate)、戊酸氯倍他索(clobetasol valerate)、地奈德(desonide)、脱氧米塞松(desoxymethasone)、乙酸脱氧皮质酮、地塞米松、二氯松(dichlorisone)、双醋二氟拉松(diflorasone diacetate)、戊酸二氟米松(diflucortolone valerate)、福阿诺龙(fluadrenolone)、氟氯奈德(fluclorolone acetonide)、氟氢可的松(fludrocortisone)、特戊酸氟地塞米松、氟轻松(fluocinolone acetonide)、乙酸氟轻松(fluocinonide)、氟可丁酯(flucortine butylester)、氟可龙(fluocortolone)、乙酸氟泼尼定(fluprednidene acetate,即乙酸氟甲叉龙(fluprednylidene acetate))、氟氢缩松(flurandrenolone)、氯氟舒松(halcinonide)、氢化可的松和其衍生物(例如乙酸氢化可的松、丁酸氢化可的松)、甲泼尼龙(methylprednisolone)、曲安奈德(triamcinoloneacetonide)、可的松(cortisone)、可托多松(cortodoxone)、乙酸氟轻松(fluocinonide)、氟氢可的松、双醋二氟拉松(diflorasone diacetate)、氟氢缩松、氟氢可的松、双醋二氟拉松、丙酮缩氟氢羟龙(flurandrenolone acetonide)、甲羟松(medrysone)、阿森纳福(amcinafel)、安息菲特(amcinafide)、倍他米松(betamethasone)和其各种酯、氯泼尼松(chloroprednisone)、乙酸氯泼尼松、氯可托龙(clocortolone)、克莱斯诺龙(clescinolone)、二氯松(dichlorisone)、二氟泼尼酯(difluprednate)、氟二氯松(flucloronide)、氟尼缩松(flunisolide)、氟甲松龙(fluoromethalone)、氟培龙(fluperolone)、氟泼尼龙(fluprednisolone)、戊酸氢化可的松、氢化可的松环戊丙酸酯、氢可他酯(hydrocortamate)、甲泼尼松(meprednisone)、帕拉米松(paramethasone)、泼尼松龙(prednisolone)、泼尼松(prednisone)、丙酸倍氯米松、曲安西龙(triamcinolone)、和其混合物。
“非类固醇消炎剂”是指一大类作用与阿司匹林(aspirin)类似的药剂,包括布洛芬(ibuprofen)(Advil)、萘普生钠(naproxen sodium)(Aleve)和醋氨芬(acetaminophen)(Tylenol)。可用非类固醇消炎剂的其它实例可包括(但不限于)昔康(oxicam),例如吡罗昔康(piroxicam)、伊索昔康(isoxicam)、替诺昔康(tenoxicam)、舒多昔康(sudoxicam)、和CP-14,304;双水杨酯(disalcid)、贝诺酯(benorylate)、三柳胆镁(trilisate)、痛热宁(safapryn)、索普林(solprin)、二氟尼柳(diflunisal)、和芬度柳(fendosal);乙酸衍生物,例如双氯芬酸(diclofenac)、芬氯酸(fenclofenac)、吲哚美辛(indomethacin)、舒林酸(sulindac)、托美丁(tolmetin)、伊索克酸(isoxepac)、呋罗芬酸(furofenac)、硫平酸(tiopinac)、齐多美辛(zidometacin)、阿西美辛(acemetacin)、芬替酸(fentiazac)、佐美酸(zomepirac)、克林酸(clindanac)、奥昔平酸(oxepinac)、联苯乙酸(felbinac)、和酮咯酸(ketorolac);芬那酯(fenamate),例如甲芬那酸(mefenamic acid)、甲氯芬那酸(meclofenamic acid)、氯芬那酸(flufenamic acid)、尼氟灭酸(niflumic acid)、和托芬那酸(tolfenamic acid);丙酸衍生物,例如布洛芬、萘普生、苯噁洛芬(benoxaprofen)、氟比洛芬(flurbiprofen)、酮洛芬(ketoprofen)、非诺洛芬(fenoprofen)、芬布芬(fenbufen)、吲哚洛芬(indoprofen)、吡洛芬(pirprofen)、卡洛芬(carprofen)、奥沙普秦(oxaprozin)、普拉洛芬(pranoprofen)、咪洛芬(miroprofen)、硫噁洛芬(tioxaprofen)、舒洛芬(suprofen)、阿明洛芬(alminoprofen)、和噻洛芬酸(tiaprofenic);吡唑,例如保泰松(phenylbutazone)、羟布宗(oxyphenbutazone)、非普拉宗(feprazone)、阿扎丙酮(azapropazone)、和三甲保泰松(trimethazone)。也可采用这些非类固醇消炎剂的混合物以及这些药剂的皮肤病学可接受的盐和酯。例如,依托芬那酯(etofenamate)(一种氯芬那酸衍生物)尤其可用于局部制剂。
本文所用“止痒剂”是指那些可减少、消除或防止瘙痒的物质。适宜止痒剂包括(但不限于)甲地嗪(methdilazine)和阿利马嗪(trimeprazine)的医药上可接受的盐。
本文所用“抗氧化剂”是指抑制氧化或抑制氧或过氧化物所促进的反应的物质。抗氧化剂的非限制性实例可包括抗坏血酸(维生素C)和其盐、脂肪酸的抗坏血酸基酯、抗坏血酸衍生物(例如抗坏血酸基磷酸镁、抗坏血酸基磷酸钠、抗坏血酸基山梨酸酯)、生育酚(维生素E)、生育酚山梨酸酯、生育酚乙酸酯、其它生育酚酯、丁基化羟基苯甲酸和其盐、6-羟基-2,5,7,8-四甲基色烷-2-甲酸(可以商品名TroloxR购得)、没食子酸和其烷基酯、尤其没食子酸丙酯、尿酸和其盐和烷基酯、山梨酸和其盐、硫辛酸、胺(例如N,N-二乙基羟胺、氨基-胍)、硫氢基化合物(例如谷胱甘肽)、二羟基富马酸和其盐、甘氨酸吡酮酸盐、精氨酸吡酮酸盐、去甲二氢愈创木酸、生物类黄酮、姜黄素、赖氨酸、甲硫氨酸、脯氨酸、超氧化物歧化酶、水飞蓟素(silymarin)、茶提取物、葡萄皮/种子提取物、黑色素、和迷迭香提取物。
“化学治疗剂”是指可用于治疗或控制疾病的化学品。化学治疗剂的非限制性实例可包括柔红霉素(daunorubicin)、多柔比星(doxorubicin)、伊达比星(idarubicin)、氨柔比星(amrubicin)、吡柔比星(pirarubicin)、表柔比星(epirubicin)、米托蒽醌(mitoxantrone)、依托泊苷(etoposide)、替尼泊苷(teniposide)、长春碱(vinblastine)、长春新碱(vincristine)、丝裂霉素C(mitomycin C)、5-FU、紫杉醇(paclitaxel)、多西他赛(docetaxel)、放线菌素D(actinomycin D)、秋水仙素(colchicine)、托泊替康(topotecan)、伊立替康(irinotecan)、吉西他滨(gemcitabine)、环孢素(cyclosporin)、维拉帕米(verapamil)、伐丝朴达(valspodar)、丙磺舒(probenecid)、MK571、GF120918、LY335979、比利考达(biricodar)、特非那定(terfenadine)、奎尼丁(quinidine)、泊维利碱A(pervilleine A)和XR9576。
本文所用“抗组胺剂”是指多种可抵抗体内组胺的化合物中的任一种,并且其可用于治疗过敏反应(例如花粉症)和感冒症状。抗组胺剂的非限制性实例可包括氯苯那敏(chlorpheniramine)、溴苯那敏(brompheniramine)、右氯苯那敏(dexchlorpheniramine)、曲普利啶(tripolidine)、氯马斯汀(clemastine)、苯海拉明(diphenhydramine)、异丙嗪(promethazine)、哌嗪、哌啶、阿司咪唑(astemizole)、氯雷他定(loratadine)和特非那定(terfenadine)。
本文所用“维生素”是指以微量对大多数动物的营养起关键作用的多种有机物质中的任一种,其尤其在代谢过程的调节中用作辅酶和辅酶前体。维生素的非限制性实例可包括维生素A和其类似物和衍生物:视黄醇、视黄醛、棕榈酸视黄酯、视黄酸、维甲酸、异维甲酸(统称为类视色素)、维生素E(生育酚和其衍生物)、维生素C(L-抗坏血酸和其酯和其它衍生物)、维生素B3(烟酰胺和其衍生物)、α-羟酸(例如羟乙酸、乳酸、酒石酸、苹果酸、柠檬酸等)和β-羟酸(例如水杨酸和诸如此类)。
本文所用“激素”是指身体器官产生的天然物质或其合成类似物,其通过血液传播以在其它位置触发活性。适宜激素可包括(但不限于)促钙醇(calciferol)(维生素D3)和其产物、雄激素、雌激素和孕酮。
本文所用“去头屑剂”是指可减少消除或预防在皮肤、尤其在头皮上形成头屑的药剂,头屑以白色或灰色小鳞片形式掉落。实例性去头屑成份可包括(但不限于)巯氧吡啶锌、页岩油和其衍生物(例如磺酸化页岩油)、二硫化硒、硫;水杨酸、煤焦油、聚维酮碘(povidone-iodine);咪唑,例如酮康唑(ketoconazole)、二氯苯基咪唑二氯戊环、克霉唑(clotrimazole)、伊曲康唑(itraconazole)、咪康唑(miconazole)、氯米巴唑(climbazole)、噻康唑(tioconazole)、硫康唑(sulconazole)、布康唑(butoconazole)、氟康唑(fluconazole)、亚硝酸咪康唑和其任何可能的立体异构体和其衍生物(例如地蒽酚(anthralin)、吡罗克酮乙醇胺(piroctone olamine)(羟甲辛吡酮(Octopirox))、二硫化硒、和环吡酮胺(ciclopiroxolamine))、和其混合物。
“抗皮肤萎缩活性剂”是指通过促进或维持天然脱皮过程来有效补充或恢复上皮层的物质。抗皱和抗皮肤萎缩活性剂的实例可包括视黄酸、其前药和其衍生物(例如顺式和反式)和类似物;水杨酸和其衍生物、含硫D和L氨基酸和其衍生物和盐,尤其N-乙酰基衍生物,其优选实例为N-乙酰基L-半胱氨酸;硫醇,例如乙硫醇;α-羟酸,例如羟乙酸、和乳酸;植酸、硫辛酸;溶血磷脂酸、和蜕皮剂(例如酚和诸如此类)。也可采用硬化剂或组织硬化剂。“组织硬化剂”是指在硬化疗法中注射至静脉中的用作化学刺激剂的试剂。最常用组织硬化剂是鱼肝油酸钠、十四烷基硫酸钠、聚乙二醇单十二醚9(laureth 9)和乙醇胺油酸酯。
可用清洁剂包括基于表面活性剂的清洁剂,其实例已列示于上文中。也可采用所属领域技术人员已知的其它并非基于表面活性剂的清洁剂。
“腐蚀剂”是指能通过化学作用破坏或侵蚀上皮组织的物质。腐蚀剂可用于移除死皮肤细胞。例如,β-羟酸是具有强角质分离效应的天然酸,其可用于问题皮肤、痤疮或起皮。
“色素淡化剂”是指能使皮肤脱色的物质。适宜色素淡化剂包括氢醌、对甲氧酚、和各种蛋白酶抑制剂,包括丝氨酸蛋白酶抑制剂、活性大豆粉(active soy)和视黄酸。
根据本发明另一方面,提供制备本文所述局部制剂的方法。所述方法一般包括混合本文所述脂化肽或肽混合物与适宜局部载剂。如果局部制剂中存在上文详述的其它活性成份,则所述方法包括将这些成份与活性成份和载剂混合在一起。本发明方法中所用的混合技术可涉及任一种调配局部组合物的已知技术。多种可用于本发明方法中的实例性调配技术阐述于(例如)以下文献中:哈氏化妆品学(Harry’s Cosmeticology),第7版,J B威尔金森(J B Wilkinson)和RJ摩尔(RJ Moore)编辑,朗曼科技(Longmann Scientific & Technical),1982。
在另一实施例中,本发明组合物必须将脂化肽或肽添加至现有(例如市售)局部组合物中。现有局部组合物可为医药组合物,在此情况下可将脂化肽或肽视作U.S.F.D.A的“新赋形剂”。新赋形剂的特征可参见由美国食品和药物管理局药物评估和研究中心在2005年5月发行的工业指导:医药赋形剂的安全评估的非临床研究(Guidance for Industry Nonclinical Studies for the Safety Evaluation of Pharmaceutical Excipients),其是以引用方式并入本文中。
根据本发明另一方面,提供治疗与上皮组织有关的医学、美容和/或药用化妆病况的方法。所述方法是通过将有效量的上述本发明组合物局部施加至表面上来实现。
本文所用术语“有效量”是指任一本发明局部制剂在投与有需要的个体(动物或人类)后可产生治疗效应或有益效应的量。所述效应可为治愈、最小化、预防或改善疾病或病症,改良身体外观和美观(例如皮肤水合),或可具有任何其它有益效应。物质的浓度经选择以发挥其医药、药用化妆或美容效应,但应低至足以避免在所属领域技术人员正确判断范围内的显著副作用。组合物的有效量可随以下因素而变:所治疗特定上皮组织、所治疗生物个体的年龄和身体状况、病况严重度、治疗持续时间、并行疗法的性质、所用具体化合物、组合物或其它活性成份、所用特定载剂、和类似因素。脂化肽的日用量一般在约0.1mg至约5g范围内变化。
可根据需要或期望局部施用局部制剂。在另一实施例中,局部制剂可一天局部施用一至四次(例如早晨一次并且晚上一次)。优选地在介于1与30天之间范围内的时间段中、更优选地在约十四天的时间段内实施本发明组合物的局部施加。某些病况可能需要在不确定时间长度中局部施加。
可将局部制剂投与有需要的个体(动物或人类)的角蛋白基质,例如上皮表面上。可施用本发明局部制剂的上皮表面的非限制性实例包括前臂的侧面、腿的侧面、肘、足、手背、背部、头皮、面部、臀部、耳道和任何其它皮肤表面、和本文所述任何粘膜。局部施加也包括将本发明组合物经口施加至牙龈。
在另一实施例中,表面是伤口表面。在慢性伤口中,局部施加可包括将局部制剂施加至非上皮表面,例如真皮。在又一实施例中,伤口表面是开放性伤口表面。本文所用“开放性伤口”是皮肤被撕裂、切破或刺破的身体创伤。本文所用“切破”是导致皮肤破裂或开放的伤害,“撕裂”是锯齿状不规则切伤,并且“刺破”是由尖锐物体(例如钉子、刀或尖牙)造成的伤口。
或者,可将组合物作为(例如)绷带、胶粘剂或透皮贴剂的组份投与上皮病况。在这些情况下,组合物可为绷带、胶粘剂或透皮贴剂的组成部分并由此使用至上皮表面。
可间接施加。例如,可将局部制剂施加至乳胶手套的内侧。在皮肤接触乳胶手套的内侧时,将本发明组合物施加至皮肤。在此实施例中,本发明组合物用于预防至少部分因包裹在手套中所致的皮肤炎症。
本文所用术语“治疗”包括消除、显著抑制、减缓或逆转病况进展、显著改善病况的临床或美观症状、显著预防外观出现病况的临床或美观症状、防止有害或扰人的刺激、或通常促进上皮组织的健康。
术语“病况”包括多种与皮肤或粘膜有关的病况。此术语意欲包括病症或疾病、促进上皮的健康、干性皮肤、和由任何潜在机制或病症所致的炎症。
本文所用“促进皮肤的健康(promotion of healthy skin或promoting healthy skin)”是指提供冷却或安抚感觉,或减少浮肿,或促使外观的起皱或浮肿减少。此词组也指使个体感知到其皮肤呈现出健康状态或产生健康或年轻感。
在另一实施例中,可将局部制剂施加至上皮组织表面以防止表面暴露于环境因素。所述因素包括(但不限于)UV辐射、风、极热天气或极冷天气。
在又一实施例中,可施用局部制剂以防止起皱。在另一实施例中,可施用局部制剂以防止光老化。在又一实施例中,可投与局部制剂以防止(例如)在尿布疹中出现的发红或浮肿。
在另一实施例中,可使用局部制剂来防止干性皮肤。可投与局部制剂以使皮肤增湿并保护皮肤免受干燥环境损害。
在另一实施例中,也可投与局部制剂以治疗已存在的皮肤病症,例如皮肤干裂。在另一实施例中,可投与局部制剂以治疗(例如)在尿布疹中出现的受刺激皮肤。
在另一实施例中,可施加局部制剂以治疗炎症。在炎症位于皮肤和粘膜中时,出现红斑(发红)并且可通过局部制剂来治疗。
众多种非限制性病况可导致炎症。这些病况包括(但不限于)a)皮炎,包括(但不限于)特应性皮炎、药物性皮炎、接触性皮炎、脂溢性皮炎、钱币状皮炎、手足部慢性皮炎、广泛性剥脱性淤滞、和局部瘙痒;b)痤疮,包括(但不限于)寻常痤疮、结囊性痤疮、暴发性痤疮、类固醇性痤疮、项部疤痕疙瘩性痤疮、氯痤疮、面部脓皮症、和囊肿;c)毛囊炎,包括(但不限于)头皮毛囊炎、泉浴泳池毛囊炎(spa pool folliculitis)、油性毛囊炎、糠秕孢子菌性毛囊炎、和革兰氏阴性菌毛囊炎(gram negative folliculitis);d)须部假毛囊炎;e)冻疮;f)痱子(汗疹);g)酒渣鼻,包括(但不限于)癣性酒渣鼻、类固醇性酒渣鼻和口周皮炎;h)湿疹和干癣;i)细菌性感染,包括(但不限于)葡萄球菌疾病、葡萄球菌烧灼性皮肤综合症、丹毒、毛囊炎、疖、痈、甲沟炎感染、和红癣;j)外科干预;k)肠病性肢端皮炎;l)斯威特氏病(Sweet’sdisease);m)淀粉样变性,包括(但不限于)淀粉样变性苔藓和斑状淀粉样变性;n)荨麻疹,包括(但不限于)急性广泛性荨麻疹和慢性广泛性荨麻疹和物理性荨麻疹;o)离心性环形红斑和环状红斑,包括(但不限于)持久性红斑、持久性回状红斑、匐行性回状红斑和持久性回状红斑;p)贝赛特氏综合征(bachet syndrome),包括(但不限于)葡萄膜炎、结节性红斑、生物素反应、皮肤病、坏疽性脓皮病、多形红斑、口疮性溃疡、肉芽肿性唇炎、疱疹样皮炎、皮肌炎(包括青少年型DM和无肌炎性DM)、嗜酸细胞性筋膜炎;q)虫咬和动物叮咬,包括(但不限于)海水浴皮疹、海藻皮炎、血吸虫皮炎、青花鱼中毒、疥疮、土风疮、和皮肤游走性幼虫症;r)真菌性感染,包括(但不限于)皮肤癣菌感染、体癣、脚癣、甲癣、头癣、股癣、花斑癣、须癣、足癣、和边缘性皮霉病;s)酵母菌感染,包括(但不限于)念珠菌病,例如白色念珠菌(Candida albicans)、口腔念珠菌(oral Candida)(鹅口疮)、念珠菌性甲沟炎;和t)寄生虫病,包括(但不限于)疥疮、虱病(包括头虱病、体虱病、和阴虱病);和v)病毒性感染,包括(但不限于)疱疹(包括单纯疱疹性病和蛇串疮)、禽痘病(水痘)、麻疹(rubeola)(麻疹(measles))和风疹(德国麻疹);w)血管舒张,包括(但不限于)雷伊氏综合征(reye’s syndrome)和伤口愈合;x)皮肤破裂创伤;y)自身免疫性病况,包括(但不限于)皮肤红斑狼疮;z)大疱性皮病,包括(但不限于)天疱疮;aa)不良药物反应;bb)免疫高反应性病况,包括(但不限于)多形性光疹、光过敏、皮肤划纹症、和多形性红斑;cc)癌症;dd)烧伤;ee)伤口;ff)囊肿;gg)化脓性汗腺炎;hh)蜂窝织炎。
尽管本文所述局部制剂不一定会治愈潜在皮肤疾病或病况,但局部制剂可用于降低或减轻皮肤疾病或病况。
另外,局部制剂可用于肛门直肠用乳膏和栓剂以治疗各种病况,例如瘙痒症、直肠炎、肛门裂、和痔。
局部制剂另外可用于眼科制剂以治疗炎症,例如由角膜溃疡、放射状角膜切开术、角膜移植、表面角膜镜片术和眼中其它手术伤口所致的炎症。
局部制剂也可以口腔洗剂或喷雾剂形式经口使用,以保护受伤口腔组织并加速其愈合,例如口疮、口腔烫伤或牙龈炎。
本文所述本发明具有人类和兽医学两种用途。本文所用术语“个体”包括禽类、爬行动物类或哺乳动物类动物。优选地,个体为哺乳动物。甚至更优选地,个体是人类。
实例
如下文实例中所述,在某些实例性实施例中,根据以下通用程序来制备肽和所述肽的调配物。应了解,尽管所述通用方法阐述了本发明某些肽和调配物的合成,但以下通用方法和所属领域技术人员已知的其它方法可适用于本文所揭示的肽和调配物中的每一者。
实例1
制备脂化肽的方法
将蛋白质提取物(100mL,5mg/ml)与胰蛋白酶(100mL,0.1mg/ml)或其它内肽酶(0.1mg/ml)混合并在37℃下培育2小时。通过外肽酶(100mL,0.1mg/ml)进一步消化提取物。在消化后,在室温下将缓冲溶液中经水解蛋白质(约4.6毫摩尔)的游离氨基与乙酸酐(10mL)一起搅拌过夜。将经保护肽转化为与2-(1H-7-氮杂苯并三唑-1-基)-1,1,3,3-四甲基脲六氟磷酸酯(HATU)(1.79g,4.6毫摩尔)的活化酯。在室温下用N,N-二异丙基乙胺(1.2g,9.2毫摩尔)使活性肽与脂化半胱氨酸残基(例如法呢基-S-半胱氨酸、植基-S-半胱氨酸、或牻牛儿基牻牛儿基-S-半胱氨酸)(4.6毫摩尔)偶合过夜,从而产生1.0g至1.8g均匀地经脂质修饰的肽的期望混合物。
实例2
制备脂化肽的方法
将蛋白质提取物(100mL,5mg/ml)与胰蛋白酶(100mL,0.1mg/ml)或其它内肽酶(0.1mg/ml)混合并在37℃下培育2小时。在室温下用DTT(770mg,4.6毫摩尔)过夜处理所得经水解肽混合物,以还原二硫键。使用诸如与伊尔曼试剂(Ellman’s reagent)(DTNB,5,5’-二硫双-(2-硝基苯甲酸))反应等方法来测量材料样品中的游离硫醇含量。使经水解蛋白提取物的样品(1ml)溶于0.1M磷酸钾缓冲液(pH 7.4)中,并在相同缓冲液中将其与20mM DTNB溶液(50μL)混合。在412nm和30℃下观察吸光度。DTNB试剂与游离硫醇反应以释放2-硝基-5-硫代苯甲酸(TNB)(1当量),其在30℃下摩尔消光系数为14,150M-1cm-1(TNB的浓度等于蛋白质样品中硫醇基的浓度)。使肽消化物(50g)悬浮于乙醇(200mL)中并添加碳酸钾(2M水溶液)以将pH调节至9.0。对于每摩尔硫醇以500μL/分钟的速率缓慢添加植基溴(1.5mol)、溴化法呢基酯、或其它活化脂质,同时剧烈搅拌以确保脂质在反应混合物中均匀分散。在添加完成后连续搅拌3小时,用冰冷盐酸(3N)将pH调节至6.0,将反应混合物蒸发至干燥,再悬浮于水(200mL)中并用三份己烷(200mL)洗涤。通过用交联葡聚糖(Sephadex)G-10管柱(在水中平衡)过滤产物来移除盐,并将期望产物蒸发至干燥。所得产物为50.0g至52.5g均匀地经脂质修饰的肽。
实例3
制备脂化肽的方法
使经氨基保护的半胱氨酸残基、N-BOC-S-法呢基-L-半胱氨酸(BOC-FC)(7.9970g,18.79mmol)溶于乙醇(37.6mL,2mL/mmol)和水(9.4mL,0.5mL/mmol)中。用碳酸铯(3mL,2M)将混合物滴定至pH 7,并蒸发溶剂以形成粗BOC-FC铯盐,然后将其溶于无水四氢呋喃(THF,各自为20mL)中,并将其蒸发三次以产生纯化BOC-FC铯盐(10.4551g)。将马里菲尔德(Merrifield)树脂(8.54g,1当量树脂:1.2当量Boc-FC)在二氯甲烷(DCM,100mL)中单独预溶胀1小时,用N,N-二甲基甲酰胺(DMF,2x100mL)冲洗DCM以产生经平衡马里菲尔德树脂。
使纯化BOC-FC铯盐溶于DMF(15mL)中,将其添加至存于125mL厄伦美尔(Erlenmeyer)烧瓶中的经平衡马里菲尔德树脂中。将反应混合物置于剧烈振荡的培育器中并在55℃下保持过夜,从而形成BOC-FC-马里菲尔德树脂。通过抽吸过滤回收BOC-FC-马里菲尔德树脂,用DMF(3x100mL)、1∶1 DMF∶水(3×100mL)、DCM(3×100mL)和甲醇(3×100mL)洗涤,并在含有氢氧化钾的真空干燥器中于20-25℃环境温度下干燥36小时,从而产生BOC-FC-马里菲尔德树脂(11.0608g)。
将BOC-FC-马里菲尔德树脂(5.6020g)置于250mL圆底烧瓶中,并且添加1∶1三氟乙酸∶DCM(100mL)。将烧瓶置于非真空旋转蒸发器上,并使反应物混合30分钟并通过抽吸过滤来移除溶剂,从而获得粗去保护FC-树脂。依序用DCM(3×100mL)、甲醇(3×100mL)、DCM(3×100mL)、甲醇(2x100mL)、戊烷(100mL)和甲醇(3×100mL)洗涤粗去保护FC-树脂,并在含有氢氧化钾的真空干燥器中干燥过夜,从而产生去保护FC-树脂(5.3364g)。
使单一经保护氨基酸(例如Boc-L-苯丙氨酸(Boc-Phe,265.3mg,1.0mmol))溶于存于250mL厄伦美尔烧瓶中并且含有HATU(418.3mg,1.1mmol)和三乙胺(415.9μL,3mmol)的DMF(2.5mL)中,并涡旋振荡5分钟以预活化氨基酸。向此混合物中添加去保护FC-树脂(1g,0.5mmol)。用铝箔盖上烧瓶并将其置于振荡培育器中并在55℃下保持过夜以形成Boc-Phe-FC-树脂。
通过抽吸过滤自反应混合物分离Boc-Phe-FC-树脂并依序用DMF(3×100mL)、DCM(4×100mL)、和THF(2×100mL)洗涤,然后使其再悬浮于氢氧化锂溶液(100mL,0.77M;3229.4mg LiOH,40mL去离子水,40mL THF,20mL甲醇)中。将反应混合物置于振荡培育器中并在55℃下保持3小时以使期望Boc-Phe-FC与树脂裂解。通过抽吸过滤移除不再固持产物的树脂,并用水(3x100mL)和乙酸乙酯(3×100mL)洗涤。用饱和氯化铵水溶液(25mL)将合并洗涤物酸化至pH 5,并用乙酸乙酯(3×100mL)来提取。用硫酸钠干燥经合并有机相并将其蒸发至干燥以产生期望Boc-苯并氨酰基-FC(254.3mg)。
实例4
调配物
基本上根据实例1中所述来制备均匀地经脂质修饰的肽。通过混合本发明的肽(50g,5%(w/v))、透明质酸钠(6250mg,0.625%(w/v))、软骨素-6-硫酸钠盐(6250mg,0.625%(w/v))、泛醇(312.5mg,0.03125%(w/v))、芦巴胶(Lubrajel)MS(1875mg,0.1875%(w/v))与甘油(1875mg,0.1875%(w/v))来制备局部血清。使混合物溶于去离子水(1L)中并在20-25℃下混合16-24小时或直至均匀溶解。
等效内容
所属领域技术人员仅使用常规实验就可了解或能确定,尽管在本文中已参照具体实施例阐述了本发明,但应理解,这些实施例仅说明本发明的原理和应用并且其它实施例可获得相同结果。因此,应理解,可对说明性实施例进行许多改变并且可设想出其它布置,而并不背离由随附权利要求书界定的本发明的精神和范畴。通过替代一般性或特别阐述的反应物和/或所用操作条件也可获得与前述实例类似的成功结果。
在权利要求书中,除非表示相反含义或上下文另有说明,否则诸如“一(a、an)”和“所述”可意指一者或不止一者。除非表示相反含义或上下文中另有说明,否则如果一个、不止一个、或所有群组成员都存于、用于、或以其它方式相关于给定产物或过程,那么可认为在群组的一或多个成员之间包括“或”的权利要求或说明适合表述这种情况。本发明包括恰好只有一个群组成员存于、用于、或以其它方式相关于给定产物或过程的实施例。本发明包括不止一个或所有群组成员都存于、用于、或以其它方式相关于给定产物或过程的实施例。此外,应理解,本发明涵盖所有变化、组合和排列,其中将一或多种限制、要素、条款、说明性术语等自一或多个所列权利要求引入另一权利要求中。例如,依赖于另一权利要求的任何权利要求均可经修改以包括在依赖于相同基础权利要求的任何其它权利要求中发现的一或多种限制。
如果以列表形式阐述要素,例如以马库西(Markush)群组格式阐述,则应了解,其也揭示所述要素的每个亚组并且可自所述群组中移除任何要素。应了解,一般来说,如果认为本发明或本发明各方面包含特定要素、特征等,那么本发明的某些实施例或本发明个方面是由所述要素、特征等组成或基本由其组成。应注意,术语“包含”意欲为开放式并且容许包括其它要素或步骤。
如果给定范围,则包括终点。此外,应了解,除非另有说明或上下文和所属领域技术人员的理解中另外明确表明,除非上下文明确表示其它含义,否则在本发明不同实施例中表示为范围的数值可表示所述范围内的任一具体值或子范围,精确到所述范围下限最小整数的十分之一。
此外,应了解,可自任何一或多项权利要求中明确排除属于先前技术范围内的任何本发明具体实施例。由于认定所述实施例是所属领域技术人员已知的,因此即使本文中没有明确阐述排除,也可将其排除。本发明组合物的任何特定实施例(例如任何靶定部分、任何疾病、病症和/或病况、任何连接剂、任何投与方法、任何治疗性应用等)都可因任何原因自任何一或多个权利要求中排除,不论其是否与现有先前技术相关。
上文和本文通篇中所讨论的公开案仅是由于其揭示内容早于本申请案的申请日期而提供。不得将本文中任何内容视作承认本发明者无权使所述揭示内容早于先前揭示内容。
Claims (76)
1.一种局部组合物,其包含含有脂化半胱氨酸残基的多肽和可接受的载剂,其中所述多肽是以占所述组合物重量0.01%至约10%的量存在。
2.如权利要求1所述的组合物,其中所述多肽的长度为约2至20个氨基酸。
3.如权利要求1所述的组合物,其中所述多肽的长度为约2至10个氨基酸。
4.如权利要求1所述的组合物,其中所述多肽的长度为约11至20个氨基酸。
5.如权利要求1所述的组合物,其中所述半胱氨酸残基经选自由以下组成的群组的脂质脂化:法呢基、牻牛儿基、植基和牻牛儿基牻牛儿基。
6.如权利要求5所述的组合物,其中所述半胱氨酸残基经法呢基脂化。
7.如权利要求1所述的组合物,其中将所述组合物施加至皮肤。
8.如权利要求1所述的组合物,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
9.如权利要求1所述的组合物,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除所述脂化半胱氨酸C-末端的至少一个氨基酸。
10.如权利要求1所述的组合物,其中所述多肽具有与选自由以下组成的群组的序列相同的C-末端氨基酸序列:SEQ ID NO 5、6、31、32、38、42、50、53、59、64、65、67、71、80、81、83、84、89、95、105和其组合。
11.如权利要求1所述的组合物,其中所述载剂是选自由以下组成的群组的口腔用载剂:漱口剂、冲洗剂、和口腔喷雾剂。
12.如权利要求11所述的组合物,其中所述口腔用载剂的pH为约5至约6.5。
13.如权利要求11所述的组合物,其中所述口腔用载剂另外包含至少一种选自由以下组成的群组的试剂:提供氟的化合物、甜味剂、着色剂、增湿剂、和乳化剂。
14.如权利要求1所述的组合物,其中所述载剂是延迟释放载剂。
15.如权利要求14所述的组合物,其中所述延迟释放载剂是选自由以下组成的群组的载剂:脂质体、微型海绵、微球体和微胶囊。
16.如权利要求1所述的组合物,其另外包含活性成份。
17.如权利要求16所述的组合物,其中所述活性成份是至少一种选自由以下组成的群组的成份:保护剂、软化剂、收敛剂、刺激剂、角质分离剂、防晒剂、仿晒剂、抗生素试剂、抗真菌剂、抗病毒剂、抗原虫剂、祛痘剂、麻醉剂、类固醇消炎剂、非类固醇消炎剂、止痒剂、抗氧化剂、化学治疗剂、抗组胺剂、维生素、激素、去头屑剂、抗皱剂、抗皮肤萎缩剂、硬化剂、清洁剂、腐蚀剂和色素淡化剂。
18.如权利要求17所述的组合物,其中所述保护剂选自由以下组成的群组:吸附剂、缓和剂和干燥剂。
19.如权利要求17所述的组合物,其中所述刺激剂是发赤剂。
20.如权利要求1所述的组合物,其另外包含类视色素。
21.如权利要求20所述的组合物,其中所述类视色素是以下物质中的至少一种:维生素A、视黄醇、视黄醛、棕榈酸视黄酯、视黄酸、维甲酸或异维甲酸或其混合物。
22.如权利要求1所述的组合物,其另外包含α-羟酸。
23.如权利要求22所述的组合物,其中所述α-羟酸是以下物质中的至少一种:羟乙酸、乳酸、酒石酸、苹果酸或柠檬酸或其混合物。
24.如权利要求1所述的组合物,其另外包含β-羟酸。
25.如权利要求1所述的组合物,其另外包含抗生素试剂。
26.如权利要求1所述的组合物,其另外包含抗真菌剂。
27.如权利要求1所述的组合物,其另外包含防晒剂。
28.如权利要求1所述的组合物,其另外包含类固醇消炎剂。
29.如权利要求1所述的组合物,其另外包含氧化钛、氧化锌、过氧苯甲酰氟尿嘧啶、间苯二酚、或水杨酸或其混合物。
30.如权利要求20至29中任一权利要求所述的组合物,其中所述多肽具有选自SEQ ID NO 1-113的C-末端氨基酸序列。
31.一种治疗上皮相关病况的方法,所述方法包含向包括人类在内的有需要的哺乳动物的表面上局部施用医药有效量的包含至少一种多肽和载剂的组合物。
32.如权利要求31所述的方法,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
33.如权利要求31所述的方法,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除脂化半胱氨酸C-末端的至少一个氨基酸。
34.如权利要求31所述的方法,其中所述多肽具有与选自由以下组成的群组的序列相同的C-末端氨基酸序列:SEQ ID NO 5、6、31、32、38、42、50、53、59、64、65、67、71、80、81、83、84、89、95、105和其组合。
35.如权利要求31所述的方法,其中所述载剂是至少一种选自基本由以下组成的群组的试剂:增湿剂、pH调节剂、除臭剂、增稠剂、增溶剂、渗透促进剂、抗刺激剂、着色剂和表面活性剂。
36.如权利要求31所述的方法,其中所述组合物中的所述载剂是选自由以下组成的群组的口腔用载剂:漱口剂、冲洗剂、和口腔喷雾剂。
37.如权利要求36所述的方法,其中所述口腔用载剂的pH为约5至约6.5。
38.如权利要求36所述的方法,其中所述口腔用载剂另外包含至少一种选自由以下组成的群组的试剂:提供氟的化合物、甜味剂、着色剂、增湿剂、和乳化剂。
39.如权利要求31所述的方法,其中所述载剂是延迟释放载剂。
40.如权利要求39所述的方法,其中所述延迟释放载剂是选自由以下组成的群组的载剂:脂质体、微型海绵、微球体和微胶囊。
41.如权利要求31所述的方法,其另外包含活性成份。
42.如权利要求41所述的方法,其中所述活性成份是至少一种选自由以下组成的群组的成份:保护剂、软化剂、收敛剂、刺激剂、角质分离剂、防晒剂、仿晒剂、抗生素试剂、抗真菌剂、抗病毒剂、抗原虫剂、祛痘剂、麻醉剂、类固醇消炎剂、非类固醇消炎剂、止痒剂、抗氧化剂、化学治疗剂、抗组胺剂、维生素、激素、去头屑剂、抗皱剂、抗皮肤萎缩剂、硬化剂、清洁剂、腐蚀剂和色素淡化剂。
43.如权利要求42所述的方法,其中所述保护剂选自由以下组成的群组:吸附剂、缓和剂和干燥剂。
44.如权利要求42所述的方法,其中所述刺激剂是发赤剂。
45.一种治疗上皮相关病况的方法,所述方法包含向包括人类在内的有需要的个体的表面上局部施用美容有效量的包含至少一种多肽和载剂的组合物。
46.如权利要求45所述的方法,其中所述表面是慢性伤口。
47.如权利要求45所述的方法,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
48.如权利要求45所述的方法,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除脂化半胱氨酸C-末端的至少一个氨基酸。
49.如权利要求45所述的方法,其中所述多肽具有与选自由以下组成的群组的序列相同的C-末端氨基酸序列:SEQ ID NO 5、6、31、32、38、42、50、53、59、64、65、67、71、80、81、83、84、89、95、105和其组合。
50.如权利要求45所述的方法,其中所述载剂是至少一种选自基本由以下组成的群组的试剂:增湿剂、pH调节剂、除臭剂、增稠剂、增溶剂、渗透促进剂、抗刺激剂、着色剂和表面活性剂。
51.如权利要求45所述的方法,其中所述组合物中的所述载剂是选自由以下组成的群组的口腔用载剂:漱口剂、冲洗剂、和口腔喷雾剂。
52.如权利要求51所述的方法,其中所述口腔用载剂的pH为约5至约6.5。
53.如权利要求51所述的方法,其中所述口腔用载剂另外包含至少一种选自由以下组成的群组的试剂:提供氟的化合物、甜味剂、着色剂、增湿剂、和乳化剂。
54.如权利要求45所述的方法,其中所述载剂是延迟释放载剂。
55.如权利要求54所述的方法,其中所述延迟释放载剂是选自由以下组成的群组的载剂:脂质体、微型海绵、微球体和微胶囊。
56.如权利要求45所述的方法,其另外包含活性成份。
57.如权利要求56所述的方法,其中所述活性成份是至少一种选自由以下组成的群组的成份:保护剂、软化剂、收敛剂、刺激剂、角质分离剂、防晒剂、仿晒剂、抗生素试剂、抗真菌剂、抗病毒剂、抗原虫剂、祛痘剂、麻醉剂、类固醇消炎剂、非类固醇消炎剂、止痒剂、抗氧化剂、化学治疗剂、抗组胺剂、维生素、激素、去头屑剂、抗皱剂、抗皮肤萎缩剂、硬化剂、清洁剂、腐蚀剂和色素淡化剂。
58.如权利要求57所述的方法,其中所述保护剂选自由以下组成的群组:吸附剂、缓和剂和干燥剂。
59.如权利要求57所述的方法,其中所述刺激剂是发赤剂。
60.一种在包括人类在内的有需要的个体中促进皮肤健康的方法,所述方法包含向包括人类在内的有需要的个体的表面上局部施用医药有效量的组合物,所述组合物包含:至少一种多肽、和载剂。
61.如权利要求60所述的方法,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
62.如权利要求60所述的方法,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除脂化半胱氨酸C-末端的至少一个氨基酸。
63.如权利要求60所述的方法,其中所述多肽具有与选自由以下组成的群组的序列相同的C-末端氨基酸序列:SEQ ID NO 5、6、31、32、38、42、50、53、59、64、65、67、71、80、81、83、84、89、95、105和其组合。
64.如权利要求60所述的方法,其中所述表面是开放性伤口。
65.如权利要求60所述的方法,其中所述表面是上皮表面。
66.如权利要求60所述的方法,其中所述表面是粘膜。
67.一种在包括人类在内的有需要的个体中促进皮肤健康的方法,所述方法包含向包括人类在内的个体的表面上局部施用美容有效量的组合物,所述组合物包含:至少一种多肽、和载剂。
68.如权利要求67所述的组合物,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
69.如权利要求67所述的组合物,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除脂化半胱氨酸C-末端的至少一个氨基酸。
70.如权利要求67所述的方法,其中所述表面是上皮表面。
71.如权利要求67所述的方法,其中所述表面是粘膜。
72.如权利要求67所述的方法,其中所述表面选自由绷带和透皮贴剂组成的群组。
73.一种制备局部组合物的方法,所述方法包含混合至少一种多肽与载剂以形成粉剂、油、乳膏、凝胶、绷带,并且其中所述局部组合物经调配以在局部施用时递送可有效抑制炎症的量的所述至少一种多肽。
74.如权利要求73所述的组合物,其中所述多肽具有选自由SEQ ID NO 1-113组成的群组的C-末端氨基酸序列。
75.如权利要求73所述的组合物,其中所述多肽具有与选自SEQ ID NO 1-113的序列相同的C-末端氨基酸序列,只是移除脂化半胱氨酸C-末端的至少一个氨基酸。
76.如权利要求73所述的方法,其中所述多肽具有与选自由以下组成的群组的序列相同的C-末端氨基酸序列:SEQ ID NO 5、6、31、32、38、42、50、53、59、64、65、67、71、80、81、83、84、89、95、105和其组合。
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US6716108P | 2008-02-25 | 2008-02-25 | |
US61/067,161 | 2008-02-25 | ||
PCT/US2009/035162 WO2009108713A2 (en) | 2008-02-25 | 2009-02-25 | Topical compositions and methods utilizing peptides containing lipid-modified cysteine-containing peptides |
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CN114729015B (zh) * | 2019-07-22 | 2024-01-12 | 第一美肤公司 | 具有抗衰老效果的多肽及其用途 |
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