CN111683653A - 用于局部递送的方法和组合物 - Google Patents
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- CN111683653A CN111683653A CN201880075223.0A CN201880075223A CN111683653A CN 111683653 A CN111683653 A CN 111683653A CN 201880075223 A CN201880075223 A CN 201880075223A CN 111683653 A CN111683653 A CN 111683653A
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Abstract
本文描述了用于局部递送活性剂的组合物和使用此类组合物的方法。组合物包括一种或多种活性剂和约0.001wt.%至约10wt.%的平均分子量为约2,000道尔顿至约20,000道尔顿的细胞外基质组分。所述细胞外组分包括透明质酸、胶原蛋白、纤连蛋白、弹性蛋白、凝集素及其片段和其组合。
Description
交叉引用
本申请要求以下各项的优先权:2017年10月11日提交的美国临时申请号62/571,025;2017年10月11日提交的美国临时申请号62/571,038;2017年12月14日提交的美国临时申请号62/598,796;2017年10月11日提交的美国临时申请号62/571,049;2017年12月14日提交的美国临时申请号62/598,786;以及2017年12月14日提交的美国临时申请号62/598,828,将其各自通过引用以其全文并入本文。
发明内容
药物施用的局部途径是可取的,因为可以避免肠胃外治疗的风险和不便;可以避免与口服治疗有关的吸收和代谢变化;药物施用可以是连续的,从而允许使用具有短生物学半衰期的药理活性剂;可以避免与许多化合物有关的胃肠刺激;相比于通过全身方法,疾病的皮肤表现可以更有效地治疗。另外,如果需要的话,局部施用活性成分对比全身施用活性成分可以在受影响区域中达到更高的组织浓度。大多数透皮递送系统和经粘膜递送系统通过使用增强渗透的媒介物或试剂来实现渗透。这样的化合物或化合物的混合物在本领域中被称为“渗透促进剂”或“皮肤促进剂”。文献中许多渗透促进剂可增强透皮吸收,但它们也具有某些缺点,因为一些缺点被认为具有毒性;一些会刺激皮肤;一些长期使用会对皮肤产生稀释(thinning)作用;而且大多数都无法递送高分子量的药物和美容剂。显然,仍然需要安全有效的透皮递送组合物和系统,这些组合物和系统可以不使用本领域已知的标准渗透促进剂而向或通过皮肤、粘膜、毛发、指甲、牙齿、骨骼和各种其他表面施用各种药物和美容剂。
本发明的各种实施例涉及包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物。
另外的实施例包括用于递送活性剂的方法,这些方法包括以下步骤:将包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物局部施用于受试者的表面组织。
在一个实施例中,提供了组合物,该组合物包含多个治疗性细胞和选自细胞外基质组分、其片段及其组合的诱饵分子。
在另一个实施例中,提供了将治疗性细胞递送至组织表面的方法,该方法包括局部施用包含多个治疗性细胞和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物。在一些实施例中,分开施用包含治疗性细胞的组合物和包含诱饵分子的组合物。
在另外的实施例中,提供了在受试者中引发免疫应答的方法,该方法包括向组织表面局部施用包含多个表达抗原的细胞和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物。在一些实施例中,分开施用包含表达抗原的细胞的组合物和包含诱饵分子的组合物。
一些实施例涉及治疗皮肤表面上的疤痕和皱纹的方法,该方法包括局部施用包含多个治疗性细胞和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物。在一些实施例中,分开施用包含治疗性细胞的组合物和包含诱饵分子的组合物。
本文还公开了可用于经由透皮贴剂递送活性剂的方法和组合物。在一个实施例中,透皮贴剂包含组合物,该组合物包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。
在另一个实施例中,提供了非侵入性地检测抗原的方法,该方法包括将透皮贴剂施加于组织表面,其中该透皮贴剂包含组合物,该组合物包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。
在另一个实施例中,提供了检测体液中的分析物的方法,该方法包括将透皮贴剂施加于组织表面以捕获体液,其中该透皮贴剂包含组合物,该组合物包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的组合物。
本发明的实施例涉及包含有效量的神经毒素剂和选自细胞外基质组分、其片段及其组合的诱饵分子的组合物。
本发明的实施例涉及向有需要的受试者局部施用神经毒素剂组合物的方法,该组合物包含有效量的神经毒素剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含神经毒素剂的组合物和包含诱饵分子的组合物。
本发明的实施例还涉及在有需要的受试者中治疗、减少或改善抬头纹(frownlines)(例如,眉间纹)、皱纹或鱼尾纹的外观的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的神经毒素剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含神经毒素剂的组合物和包含诱饵分子的组合物。
本发明的实施例涉及治疗有需要的受试者的出汗症状的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的神经毒素剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含神经毒素剂的组合物和包含诱饵分子的组合物。
本发明的实施例涉及在有需要的受试者中治疗偏头痛的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的神经毒素剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含神经毒素剂的组合物和包含诱饵分子的组合物。
本发明的实施例涉及通过向有需要的受试者局部施用有效量的一种或多种活性剂和选自透明质酸酶、弹性蛋白酶或其组合的酶来治疗该受试者的病症的方法。在一些实施例中,组合物进一步包含选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含活性剂的组合物和包含透明质酸酶/弹性蛋白酶的组合物。
本发明的实施例涉及通过向皮肤表面局部施用组合物来治疗或减少有需要的受试者的皮肤表面上的疤痕和皱纹的方法,该组合物包含有效量的一种或多种活性剂和选自透明质酸酶、弹性蛋白酶或其组合的酶。在一些实施例中,组合物进一步包含选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含活性剂的组合物和包含透明质酸酶/弹性蛋白酶的组合物。
本发明的实施例涉及通过向有需要的受试者局部施用组合物来治疗该受试者的脱发的方法,该组合物包含有效量的一种或多种活性剂和选自透明质酸酶、弹性蛋白酶或其组合的酶。在一些实施例中,组合物进一步包含选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含活性剂的组合物和包含透明质酸酶/弹性蛋白酶的组合物。
本发明的实施例涉及通过向有需要的受试者局部施用组合物来治疗该受试者的皮肤病症的方法,该组合物包含有效量的一种或多种活性剂和选自透明质酸酶、弹性蛋白酶或其组合的酶。在一些实施例中,组合物进一步包含选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,分开施用包含活性剂的组合物和包含透明质酸酶/弹性蛋白酶的组合物。
附图说明
为了更充分理解本发明的性质和优势,应该结合以下附图来参考以下详细说明,其中:
图1A-1B是显示针对包含透明质酸诱饵分子(分子量为10,000Da、20,000Da、40,000Da、60,000Da或100,000Da)的肽组合物的肽通量相对于来自单独肽组合物的肽通量的百分比的图,其中在具有完整角质层的皮肤中(图1A)和在具有剥离角质层的皮肤中(图1B)测量通量,并且每种组合物以一式两份进行测量(实线、虚线)。
图2是柱状图,其显示了相比不含诱饵分子的组合物,来自水杨酸盐和透明质酸诱饵分子(分子量指定为小(5,000Da至10,000Da)、小至中(10,000Da至20,000Da)、低至中(20,000Da至30,000Da)和中(30,000Da至40,000Da))的组合物的水杨酸盐通量的百分比增加。
图3是柱状图,其显示了相比不含诱饵分子的组合物,来自氢化可的松和透明质酸诱饵分子(分子量指定为非常小(5,000Da至10,000Da)、小(10,000Da至20,000Da)、中(30,000Da至40,000Da)和大(40,000Da至60,000Da))的组合物的氢化可的松通量的百分比增加。
图4是柱状图,其显示了来自利多卡因和弹性蛋白诱饵分子(分子量指定为非常非常小(2,000Da至5,000Da)、非常小(5,000Da至10,000Da)、以及小(10,000Da至20,000Da))的局部施用组合物的利多卡因以及来自无诱饵分子的组合物的利多卡因在猪皮肤中的百分比。
图5是柱状图,其显示了与不含诱饵的组合物相比,来自含有米诺环素和透明质酸诱饵分子(分子量指定为3,000Da、5,000Da和10,000Da)的组合物的局部施用的米诺环素在猪皮肤中的百分比。
图6是显示UVA和UVB在皮肤中的吸收的柱状图(4.0对应于100%),其中这些柱对应于具有添加到市售防晒剂(Anthelios 60)中的诱饵分子的防晒组合物、和市售防晒剂(Anthelios 60)。
图7A-7B是针对单独的市售防晒剂(Anthelios 60)(图7A)和针对具有诱饵分子的市售防晒剂(Anthelios 60)(图7B)的UV吸收与波长(nm)之间的关系的图。
图8是显示针对市售防晒剂(Anthelios 60)(实线)和针对具有诱饵分子的市售防晒剂(Anthelios 60)(虚线)经皮肤的UV吸收百分比与波长(nm)之间的关系的图。
图9是柱状图,其显示了递送到体外猪皮肤移植物中的、来自加巴喷丁和透明质酸钠的局部配制品以及来自仅加巴喷丁的局部配制品的加巴喷丁在组织中的量(μg加巴喷丁/g组织)。
图10是柱状图,其显示了递送到体外猪皮肤移植物中的、来自pal-KTTKS和透明质酸钠的局部配制品以及来自仅pal-KTTKS的局部配制品的棕榈酰-赖氨酸-苏氨酸-苏氨酸-赖氨酸-丝氨酸(pal-KTTKS)在组织中的量(μg pal-KTTKS/50mg组织)。
图11是柱状图,其显示了与水杨酸盐和盐水的组合物相比,当组合物中包括弹性蛋白诱饵分子时,水杨酸盐在猪粘膜组织中的递送的百分比增加。
图12是柱状图,其显示了与单独的抗体相比,来自包含抗体和透明质酸诱饵分子(分子量指定为非常非常低、非常低和低)的组合物的抗体通量的百分比增加。
图13显示了在诱饵分子存在下,施加至眉间区的FITC-右旋糖酐的共聚焦成像。
图14显示了通过共聚焦成像,FITC-右旋糖酐随时间在眉间区中的渗透。
图15显示了3hr后在多种诱饵分子ILS-20(A)和ILS-3(B)存在下,FITC-右旋糖酐在外侧眼角区域中的渗透。
图16描绘了在基线时肉毒杆菌毒素(botox)的局部施用。
图17描绘了在第1周和第8周,局部施用的根据本文所述实施例的肉毒杆菌毒素+ILS-20对腋窝汗液的作用。
图18描绘了在第1周和第8周,局部施用的根据本文所述实施例的肉毒杆菌毒素+ILS-3对腋窝汗液的作用。
具体实施方式
现在将在下文中更全面地描述各个方面。然而,这些方面能以许多不同的形式呈现并且不应被解释为限于本文示出的实施例;更确切地说,提供这些实施例,以便本公开是全面的和完整的,并将其范围充分地传达给本领域技术人员。
在提供值的范围时,意在该范围和任何其他阐述的范围的上限与下限之间的每个中间值或在该阐述范围内的中间值均被涵盖在本公开之内。例如,如果阐述的范围是1μm至8μm,则意在也明确公开2μm、3μm、4μm、5μm、6μm和7μm、以及大于或等于1μm的值的范围且小于或等于8μm的值的范围。
单数形式“一个/一种(a/an)”和“该(the)”包括复数个指示物,除非上下文中另外明确指明。因此,例如,提及一种“聚合物”包括单一聚合物以及两种或更多种相同或不同的聚合物;提及一种“赋形剂”包括单一赋形剂以及两种或更多种相同或不同的赋形剂,等等。
所有百分比、份数和比率都是基于局部组合物的总重量,并且在约25℃进行所有测量,除非另有说明。
紧接数值之前的单词“约”表示该值的加或间10%的范围,例如,“约50”表示45至55,“约25,000”表示22,500至27,500,等等,除非本公开的上下文另外指明或与这种解释不一致。例如,在如“约49、约50、约55”的数值列表中,“约50”意指范围扩展到小于先前值与后续值之间的间隔的一半,例如,大于49.5至小于52.5。此外,应根据本文提供的术语“约”的定义来理解短语“小于约”一个值或“大于约”一个值。
如本文所用的术语“施用(administer/administering/administration)”是指直接施用化合物(也称为目的试剂)或该化合物(目的试剂)的药学上可接受的盐或组合物。
术语“抗原”可以是肽、多肽、蛋白质、糖蛋白、脂蛋白、脂质、磷脂、碳水化合物、糖脂、其混合物或缀合物、或已知诱导免疫的任何其他材料。抗原的分子量可以大于1千道尔顿(kDa)、10kDa或100kDa(包括其中间范围)。抗原可以与载体缀合。抗原可以作为完整的生物体提供,例如细菌或病毒体;抗原可以从生物体的提取物或裂解物中获得,例如从全细胞或从膜中获得;抗原可以作为活生物体(例如像活病毒或细菌)、减毒活生物体(例如像减毒活病毒或细菌)、或通过化学或遗传技术灭活的生物体提供;并且抗原可以化学合成、通过重组技术生产或从天然来源纯化。
术语“抗体片段”可以包括小于全长的抗体的任何衍生物。在示例性实施例中,抗体片段保留了全长抗体的特异性结合能力的至少重要部分。抗体片段的实例包括但不限于Fab、Fab'、F(ab')、scFv、Fv、双抗体、三抗体、四抗体、Fd片段或其混合物。抗体片段可以通过任何方式产生。例如,抗体片段可以通过完整抗体的片段化而酶促或化学地产生,可以由编码部分抗体序列的基因重组产生,或者可以全部或部分地合成产生。抗体片段可以任选地是单链抗体片段。可替代地,该片段可以包含多个链,这些链例如通过二硫键连接在一起。该片段还可以任选地是多分子复合物。
与“包括(including)”、“含有(containing)”或“特征在于(characterized by)”同义的过渡性术语“包含(comprising)”是包含性的或开放性的并且不排除附加的、未列举的元素或方法步骤。相比之下,过渡性短语“由……组成”不包括未在权利要求中指明的任何元素、步骤或成分。过渡性短语“基本上由……组成”将权利要求的范围限制于指定的材料或步骤以及“不实质地影响所要求保护的发明的一个或多个基础且新颖的特征的那些”。在实施例或权利要求中,其中术语包含被用作过渡性短语,用术语“由……组成”或“基本上由……组成”替换术语“包含”也可以被设想为此类实施例。本公开的组合物和方法可以包含所公开的组分、基本上由其组成或由其组成。
如本文所用的术语“载体”涵盖载体、赋形剂和稀释剂,意指材料、组合物或媒介物,例如液体或固体填充剂、稀释剂、赋形剂、溶剂、或包封材料(该包封材料参与携带或运输药物、化妆品或其他药剂跨越组织层(例如角质层或棘层))。
除非另有指示,否则本公开使用术语“障碍”意指术语疾病、病症、或病痛,并且术语“障碍”可与这些术语互换地使用。
如本文所用的术语“诱饵分子”可与如本文所述的细胞外基质组分、其片段及其组合互换。
术语“有效量”和“治疗有效量”在本公开中可互换使用,并且是指当组合物、化合物或活性剂施用于受试者时能够减轻患者中障碍的症状或增强预期组织治疗区域的肌理、外观、颜色、感觉或水合作用。包含“有效量”或“治疗有效量”的实际量将根据许多条件而变化,包括但不限于障碍的严重程度、患者的大小和健康状况以及施用途径。有技术的执业医师可以使用医学领域已知的方法容易地确定合适的量。
本文使用短语“药学上可接受的”是指在合理医学判断范围内的那些目的药剂/化合物、盐、组合物、剂型等,它们适用于与人类和/或其他哺乳动物的组织接触而没有过多的毒性、刺激、过敏反应或其他问题或并发症,且与合理的益处/风险比相称。在一些方面,“药学上可接受的”意指由联邦或州政府的监管机构批准或者列于美国药典(U.S.Pharmacopeia)或其他普遍认可的适用于哺乳动物(例如动物)并且更特别地适用于人的药典中。
如本文所用的术语“盐”包括通常用于形成游离酸的碱金属盐和形成游离碱的加成盐的药学上可接受的盐。该盐的性质不是关键的,条件是它是药学上可接受的。术语“盐”还包括加成盐的溶剂化物,例如水合物、以及加成盐的多晶型物。合适的药学上可接受的酸加成盐可以由无机酸或有机酸制备。此类无机酸的非限制性实例为盐酸、氢溴酸、氢碘酸、硝酸、碳酸、硫酸和磷酸。适当的有机酸可以选自脂肪族酸、脂环族酸、芳香族酸、芳基脂肪族酸、以及含有杂环基的羧酸、以及磺酸,例如甲酸、乙酸、丙酸、琥珀酸、乙醇酸、葡糖酸、乳酸、苹果酸、酒石酸、柠檬酸、抗坏血酸、葡糖醛酸、马来酸、富马酸、丙酮酸、天冬氨酸、谷氨酸、苯甲酸、邻氨基苯甲酸、甲磺酸(mesylic acid)、硬脂酸、水杨酸、对羟基苯甲酸、苯乙酸、扁桃酸、双羟萘酸(帕莫酸)、甲磺酸、乙磺酸、苯磺酸、泛酸、甲苯磺酸、2-羟基乙磺酸、磺胺酸、环己基氨基磺酸、海藻酸、3-羟基丁酸、半乳糖二酸以及半乳糖醛酸。
术语“患者”和“受试者”可互换,并且可以被认为意指可以用本发明的化合物治疗的任何活的生物体。同样,术语“患者”和“受试者”可以包括但不限于任何非人类哺乳动物、灵长类动物或人类。在一些实施例中,“患者”或“受试者”是哺乳动物,例如小鼠、大鼠、其他啮齿动物、兔、犬、猫、猪、牛、绵羊、马、灵长类动物或人类。在一些实施例中,患者或受试者是成人、儿童或婴儿。在一些实施例中,患者或受试者是人类。
在本文中,例如提及治疗皮肤障碍或全身性病症的方法时使用术语“治疗”,并且该术语通常包括施用化合物或组合物,相对于未接受该化合物或组合物的受试者而言,该化合物或组合物可降低医学病症的症状的频率或延迟其发作,或可改善受试者的组织表面的预期组织治疗区域的肌理、外观、颜色、感觉或水合作用。这可以包括以改善或稳定受试者的病症的方式逆转、减少、或抑制病症的症状、临床征象、和潜在病理。
通过保留提供或排除任何此类组(可以根据范围或以任何类似方式要求保护,包括该组内的任何子范围或子范围的组合)的任何单个成员的权利,可以出于任何原因要求保护少于本公开的全部测量值。此外,通过保留提供或排除任何单独的取代基、类似物、化合物、配体、结构或其基团、或所要求保护的基团的任何成员的权利,可以出于任何原因要求保护少于本公开的全部测量值。在整个本公开中,参考了各种专利、专利申请和出版物。这些专利、专利申请和出版物的公开内容通过引用以其全文并入本公开中,以便更全面地描述截至本公开日期的本领域技术人员已知的现有技术水平。在引用的专利、专利申请和出版物与本公开之间存在任何不一致的情况时,将以本公开为准。
为了方便起见,在此收集了说明书、实例和权利要求书中采用的某些术语。除非另外定义,否则在本公开中所用的全部技术和科学术语具有与本公开所属领域的普通技术人员通常所理解的相同的意义。
本发明的各种实施例涉及用于局部递送活性剂的组合物。这些组合物包含活性剂和诱饵分子,该诱饵分子能够通过暂时破坏细胞-细胞(即,细胞间)和细胞-支架的附着而引起组合物接触的组织的重排,从而使活性剂穿过细胞层并被动地分布于整个组织中。本文所述的组合物和方法可用于施用任何活性剂,包括小分子药物、大分子药物、生物制剂、抗体、嵌合抗体、抗原、肽、抗氧化剂、化妆品成分、治疗性细胞、诊断剂、放射性示踪剂、造影剂、神经毒素等及其组合。这些组合物和方法还可以用于诊断目的和介导诊断性分子流经各种组织。在一些实施例中,可以将这些组合物施用于任何表面组织,包括皮肤、粘膜、眼睛、耳朵、鼻子内部、口腔内部、嘴唇、尿道开口、阴道、肛门、舌头、舌系带、毛发、牙齿、骨骼、泪腺、鼻窦粘膜、呼吸道、牙龈等及其组合。
诱饵分子
在某些实施例中,诱饵分子可以是细胞外基质组分或其片段。基于细胞外基质组分的结构及其在细胞外基质中的功能,可将其分为几种类别。最突出的类别是细胞外基质蛋白的结构类别。这些主要由胶原蛋白和弹性蛋白家族组成。胶原蛋白纤维使基质增强并使基质组织起来;弹性蛋白纤维提供柔韧性和回弹性。另一种类别是特化蛋白质,例如原纤蛋白、纤连蛋白、层粘连蛋白、分区蛋白、生腱蛋白和玻连蛋白在细胞外基质中起较少的结构作用,而起较多的粘着或整合作用;这些蛋白质允许细胞附着并在基质凝胶内形成交联。最后,许多蛋白聚糖和含有硫酸乙酰肝素的蛋白质形成了高度水合的凝胶状混合物,该混合物有助于将基质稳定在其水性环境中。蛋白聚糖由蛋白质核心组成,在该蛋白质核心上附着了糖胺聚糖(GAG)的长链,形成了ECM的极其复杂的高分子量组分。作为细胞外基质组分的另一种GAG是透明质酸,一种非硫酸盐GAG。
例如,在一些实施例中,诱饵分子可以是蛋白质、肽、或与细胞外基质相关的受体、透明质酸、弹性蛋白、胶原蛋白、纤连蛋白、凝集素及其片段和其组合。此类片段包括但不限于透明质酸片段、胶原蛋白片段、纤连蛋白片段、弹性蛋白片段、凝集素片段及其组合。
在特定的实施例中,诱饵分子可以是透明质酸。已知透明质酸与例如CD44(透明质酸介导的运动的受体(RHAMM))和细胞间粘附分子-1(ICAM-1)相互作用。CD44广泛分布于全身,并介导与透明质酸的细胞相互作用。ICAM-1是透明质酸的代谢细胞表面受体,并且透明质酸与ICAM-1的结合可能有助于控制ICAM-1介导的炎症激活。透明质酸是二糖的聚合物。不希望受理论束缚,透明质酸的低分子量片段可通过中断细胞间相互作用和/或触发细胞损伤反应来破坏细胞-细胞和细胞-支架的附着,这可能破坏不直接接触透明质酸诱饵分子的细胞之间的细胞间相互作用。透明质酸可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。在一些实施例中,透明质酸片段不是交联的。
在一些实施例中,诱饵分子可以是胶原蛋白。胶原蛋白能以多种形式从多种来源中分离出来。示例性胶原蛋白包括I型胶原蛋白、II型胶原蛋白、III型胶原蛋白、IV型胶原蛋白或V型胶原蛋白。胶原蛋白还可以是纤维胶原蛋白或非纤维胶原蛋白。低分子量胶原蛋白可以例如通过水解来制备,并且像透明质酸一样,低分子量胶原蛋白可以通过中断细胞间相互作用和/或触发细胞损伤反应来破坏细胞-细胞和细胞-支架的附着,这可能会破坏组织更深处细胞之间的细胞间相互作用。胶原蛋白片段可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
在某些实施例中,诱饵分子可以是纤连蛋白。纤连蛋白是蛋白质二聚体,由通过一对二硫键连接的两个几乎相同的单体组成。纤连蛋白与称为整合素的跨膜受体蛋白和细胞外基质组分(例如胶原蛋白、纤维蛋白和硫酸肝素蛋白聚糖)结合。像透明质酸和胶原蛋白一样,纤连蛋白片段可以通过中断细胞间相互作用和/或触发细胞损伤反应来破坏细胞-细胞和细胞-支架的附着,这可能会破坏组织更深处细胞之间的细胞间相互作用。纤连蛋白片段可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
在一些实施例中,诱饵分子可以是弹性蛋白。弹性蛋白是结缔组织中发现的一种蛋白质,可使人体许多组织在拉伸或收缩后恢复其形状。像透明质酸、胶原蛋白和纤连蛋白一样,弹性蛋白片段可以通过中断细胞间相互作用和/或触发细胞损伤反应来破坏细胞-细胞和细胞-支架的附着,这可能会破坏组织更深处细胞之间的细胞间相互作用。弹性蛋白片段可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
弹性蛋白片段可商购获得或可通过蛋白酶消化产生,例如使用蛋白酶K或嗜热菌蛋白酶。例如,可从密苏里州圣路易斯的蛋白质制剂公司(Protein Preparations,Inc.,St.Louis,MO)商购的弹性蛋白E91制剂是适合进行消化的弹性蛋白产物,其分子量为约1,000至约60,000道尔顿。另外,还可以使用以商品名ProK(特别是ProK-60和ProK-60P)可获得的一系列消化物,它们是由源自牛颈部韧带的不溶性弹性蛋白的蛋白水解消化得到的弹性蛋白肽混合物。
在一些实施例中,这些弹性蛋白肽片段可以包含选自由以下组成的组的氨基酸序列:GAAPG、GVVPG、GGGPG、GLLPG、GIIPG、GSSPG、GTTPG、GCCPG、GMMPG、GFFPG、GYYPG、GWWPG、GDDPG、GNNPG、GEEPG、GQQPG、GRRPG、GHHPG、GKKPG、GPPPG、G3Hyp3HypPG(甘氨酸-3-羟基脯氨酸-3-羟基脯氨酸-脯氨酸-甘氨酸)、G4Hyp4HypPG(甘氨酸-4-羟基脯氨酸-4-羟基脯氨酸-脯氨酸-甘氨酸)、RRPEV、QPSQPGGV、PGGV、GPGV、KPGV、GPGL、EGSA、PGGF、GGGA、KPGKV、PGGV、KPKA、GPGGV、GPQA、GGPGI、PGPGA、GPGGV、GQPF、GGKPPKPF、GGQQPGL、GGPGI、VGVAPG、IGVAPG、PGGVLPG、VGVVPG、IGLGPGGV、VGAMPG、VGLSPG、IGAMPG、IGLSPG、GVAPGV、VAPGVG、APGVGV、PGVGVA、以及GVGVAP。
在一些实施例中,诱饵分子可以是层粘连蛋白。层粘连蛋白是一种复合物,其由三个不同的多肽链(α、β、γ)组成,这些多肽链通过二硫键彼此结合成十字形分子,该十字形分子包含每端均带有小球的一个长臂和三个短臂。α-2链是层粘连蛋白-2(分区蛋白)和层粘连蛋白-4(S-分区蛋白)的亚基。它的细胞结合能力(经由膜结合的整合素受体)使层粘连蛋白成为刺激和增强细胞迁移和神经突增生的有效底物涂层。在来自胎盘的层粘连蛋白中,A链被分区蛋白替代,而在神经肌肉接点附近发现的层粘连蛋白中,B1链被s-层粘连蛋白(突触层粘连蛋白)替代。层粘连蛋白片段可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
在一些实施例中,诱饵分子可以是凝集素。凝集素通常是复杂的多结构域多聚体蛋白。但是,哺乳动物凝集素的碳水化合物结合活性通常是碳水化合物识别结构域或CRD的特性。哺乳动物凝集素的CRD分为三个系统发育保守的类别:C型、S型和P型。C型凝集素需要Ca++才能与配体结合,是细胞外膜和可溶性蛋白质,并且这一类别可以结合多种碳水化合物。S型凝集素在还原条件下最具活性,在细胞内和细胞外均存在,结合β-半乳糖苷并且不需要Ca++。P型凝集素结合作为其主要配体的甘露糖6-磷酸盐。凝集素片段可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
在一些实施例中,诱饵分子可以是以下至少一种:硫酸肝素、硫酸软骨素、硫酸角质素、层粘连蛋白、分区蛋白、生腱蛋白、玻连蛋白和原纤蛋白、及其片段和其组合。这些诱饵分子可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
在一些实施例中,诱饵分子可以是细胞外基质受体,例如整合素、盘状结构域受体和多配体聚糖、及其片段和组合。这些诱饵分子可以具有的平均分子量为小于约60,000Da、约2,000Da至约60,000Da、约2,000Da至约40,000Da,约5,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000Da至约20,000Da、或约2,000Da至约10,000Da。
诱饵分子的大小可以影响细胞-细胞和细胞支架的破坏,并且在各种实施例中,诱饵分子可以具有的平均分子量为约2000道尔顿至约100,000道尔顿(“Da”)。在特定的实施例中,诱饵分子可以具有的平均分子量为约2,000Da至约60,000、约2,000Da至约40,000Da、或约5,000Da至约40,000Da。在其他实施例中,诱饵分子可以具有的平均分子量为约2,000Da至约5,000Da(“非常小”尺寸)、约5,000Da至约10,000Da(“小”尺寸)、约10,000Da至约20,000Da(“小至中”尺寸)、约20,000Da至约30,000Da(“低至中”尺寸)、约30,000Da至约40,000Da(“中”尺寸)、约40,000Da至约60,000Da(“大”尺寸)、或约60,000Da至约100,000Da(“非常大”尺寸)。因为诱饵分子通常包括细胞外基质组分的片段,所以实施例的组合物可以包括落在指定的“平均分子量”之上和之外的任何范围内的诱饵分子。例如,当平均分子量为小至中时,诱饵分子可以包括大和超大或非常小和小的单个分子。
在实施例中,组合物中存在的细胞外基质组分、其片段及其组合(诱饵分子)具有指定的平均分子量。在实施例中,诱饵分子可以具有的平均分子量为约2,000Da至约100,000Da、约2,000Da至约60,000Da、约2,000Da至约50,000Da、约2,000Da至约40,000Da、约2,000Da至约30,000Da、约2,000至约20,000Da、约2,000至约15,000Da、约2,000Da至约10,000Da、约5,000Da至约40,000Da、小于约60,000Da、小于约50,000Da、小于约40,000Da、小于约30,000Da、小于约20,000Da、小于约15,000Da、小于约10,000Da、小于约5,000Da、约60,000Da、约50,000Da、约40,000Da、约30,000Da、约20,000Da、约15,000Da、约12,500Da、约10,000Da、约8,500Da、约7,500Da、约5,000Da、约2,000Da至约5,000Da、约5,000Da至约10,000Da、约10,000Da至约20,000Da、约20,000Da至约30,000Da、约30,000Da至约40,000Da、约20,000Da至约40,000Da、约40,000Da至约60,000Da、或约60,000Da至约100,000Da、或落入这些实例范围和数字内的任何范围或单个数字。
在实施例中,这些组合物可基本上不包括分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的细胞外基质组分、其片段及其组合(诱饵分子)。在其他实施例中,该组合物可以不含有分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的可检测的诱饵分子。在另外的实施例中,该组合物可以含有小于0.05%(即,小于组合物中存在的总诱饵分子的0.05wt.%)的分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的诱饵分子。在另外的实施例中,该组合物可以含有小于0.1%(即,小于组合物中存在的总诱饵分子的0.1wt.%)的分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的诱饵分子。在一些实施例中,该组合物可以含有小于1%(即,小于组合物中存在的总诱饵分子的1wt.%)的分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的诱饵分子。
在实施例中,诱饵分子是透明质酸或其片段,其中基本上所有的透明质酸或其片段均不具有约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的分子量。在其他实施例中,诱饵分子是透明质酸或其片段,其中不存在分子量约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上的可检测的透明质酸或片段。在其他实施例中,诱饵分子是透明质酸或其片段,其中小于0.1%的透明质酸或其片段的分子量分子量为约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上。在其他实施例中,诱饵分子是透明质酸或其片段,其中小于1%的透明质酸或其片段的分子量分子量为约150,000Da以上、约125,000Da以上、约100,000Da以上、约90,000Da以上、约80,000Da以上、约70,000Da以上、60,000Da以上、约55,000Da以上、约50,000Da以上、约45,000Da以上、约40,000Da以上或约35,000Da以上。在实施例中,诱饵分子可以进一步包含胶原蛋白、纤连蛋白、弹性蛋白、凝集素、胶原蛋白片段、纤连蛋白片段、弹性蛋白片段、凝集素片段及其组合。
组合物中诱饵的量可通过调节破坏的深度来影响细胞-细胞和细胞-支架的破坏,从而调节活性剂的渗透深度。通常,各种实施例的组合物中存在的诱饵的量可以为约0.001wt.%至约10wt.%,并且在特定实施例中,在这样的组合物中诱饵的量可以为约0.1wt.%至约2.0wt.%、约0.25wt.%至约3.0wt.%、约0.5wt.%至约5.0wt.%、约0.75wt.%至约7.5wt.%、或涵盖这些实例范围的任何范围或单个浓度。如上所示,诱饵分子的量可以调节活性剂的渗透深度。例如,当局部施用组合物时,诱饵分子的相对较低的浓度(例如约0.1wt.%至约2.0wt.%、或约0.25wt.%至约1.0wt.%)可以允许活性剂部分地穿过表皮(例如,穿过颗粒层)转运并进入棘层。当局部施用该组合物时,诱饵分子的较高浓度(例如约0.5wt.%至约5.0wt.%、或约0.5wt.%至约3.0wt.%)可以允许活性剂完全穿过表皮转运至组织层下面的基底膜,例如真皮、皮下组织和血流。相对于组合物的总量,本文公开的重量百分比可以是重量/重量百分比或重量/体积百分比。
在一些实施例中,一种或多种诱饵分子能以总组合物的约0.001wt.%至约10wt.%、约0.001wt.%至约9wt.%、约0.001wt.%至约8wt.%、约0.001wt.%至约7wt.%、约0.001wt.%至约6wt.%、约0.001wt.%至约5wt.%、约0.001wt.%至约4wt.%、约0.001wt.%至约3wt.%、约0.001wt.%至约3wt.%、或约0.001wt.%至约1wt.%存在。具体实例包括约0.001wt.%、约0.01wt.%、约0.1wt.%、约0.5wt.%、约1wt.%、约2wt.%、约5wt.%、约10wt.%、以及介于这些值中的任何两个值之间的范围。相对于组合物的总量,本文公开的重量百分比可以是重量/重量百分比或重量/体积百分比。
在一些实施例中,一种或多种诱饵分子能以约1微克至约100毫克/mL组合物、约1微克至约10毫克/mL组合物、约1微克至约5毫克/mL组合物、约1微克至约1毫克/mL组合物、或约1微克至约100微克/mL组合物存在。
因为诱饵分子的浓度可以调节活性剂的渗透深度,所以靶向例如表皮的活性剂可以包括在含有更低浓度诱饵分子(例如,约0.1wt.%至约2.0wt.%、或约0.25wt.%至约1.5wt.%)的组合物中,并且靶向例如真皮或皮下组织的活性剂可以包括在含有较高浓度诱饵分子(例如,约1.0wt.%至约5.0wt.%、或约1.0wt.%至约3.0wt.%)的组合物中。类似地,活性剂的大小可影响组合物的配制品。例如,大的活性剂(例如大分子治疗性剂或生物性/治疗性肽)可能需要更高浓度的诱饵分子(例如约0.5wt.%至约5.0wt.%、或约0.5wt.%至约3.0wt.%),以允许向表皮施用,尽管相似的浓度可以允许较小的治疗剂施用至真皮或通过血流全身施用。
透明质酸酶/弹性蛋白酶
在一些实施例中,可以使用例如透明质酸酶和弹性蛋白酶等酶代替诱饵分子或与诱饵分子组合以促进活性剂的递送。不希望受到理论束缚,透明质酸酶和/或弹性蛋白酶的存在可能通过降解细胞外基质并暂时破坏细胞-细胞(即,细胞间)和细胞-支架的附着而引起组织重排,并允许活性剂有效地穿过细胞层进入组织。当施用时,透明质酸酶和弹性蛋白酶可以在组织中迅速传播和扩散,可以通过水解透明质酸和弹性蛋白来改变细胞间粘合质的渗透性和粘度。
在一些实施例中,透明质酸酶和弹性蛋白酶可以与诱饵分子组合使用。诱饵分子的存在也可能破坏细胞-细胞(即,细胞间)和细胞-支架的附着,从而协同地增强透明质酸酶和弹性蛋白酶的作用,并允许活性剂渗透并穿过组织。诱饵分子可与透明质酸酶/弹性蛋白酶一起提供协同作用,并更有效地递送活性剂。
在实施例中,透明质酸酶和弹性蛋白酶可以在施用活性剂和诱饵分子的延迟期之后导致活性剂渗透或扩散到下面的组织中。高分子量诱饵分子的存在可能不会有效地导致活性剂渗透并穿过组织。在延迟期期间,组合物中的透明质酸酶和弹性蛋白酶可降解高分子量诱饵分子并产生更小的片段,这些片段可有助于活性剂的渗透。因此,透明质酸酶和弹性蛋白酶、一种或多种诱饵分子的选择、平均分子量、高分子量诱饵分子的存在(或不存在)、诱饵分子的量、递送的活性剂和靶标表面组织将影响活性剂局部递送至所需作用部位的能力。
在实施例中,这些组合物可包括选自透明质酸酶、弹性蛋白酶的酶或其组合。透明质酸酶家族由能够水解或“分解”多糖透明质酸的酶组成。透明质酸是结缔组织的重要成分。透明质酸酶可概括地分为三类:哺乳动物型透明质酸酶(EC 3.2.1.35)是内切-β-N-乙酰基己糖胺酶,其生产四糖和六糖作为主要终产物。它们具有水解活性和转糖苷酶活性,并且可以降解透明质酸和硫酸软骨素(CS),特别是C4-S和C6-S。细菌透明质酸酶(EC4.2.99.1)降解透明质酸,并且在不同程度上降解CS和DS。它们是内切-β-N-乙酰基己糖胺酶,通过β消除反应起作用,主要产生二糖终产物。来自水蛭、其他寄生虫和甲壳类动物的透明质酸酶(EC 3.2.1.36)是内切-β-葡糖醛酸糖苷酶,通过β1-3键的水解产生四糖和六糖终产物。
本文公开的透明质酸酶可以源自任何来源,例如可以从牛蛋白(牛型)、水蛭或细菌(例如,呈透明质酸裂合酶的形式)中回收。透明质酸酶还可以是植物来源的。本领域的基因工程技术同样可以用于产生透明质酸酶。各种类型的透明质酸酶也可商购获得,例如,购自惠氏药厂(Wyeth-Ayerst)雅培公司(Abbot)(Hyazyme)、百时美施贵宝公司(Bristol-Myers Squibb)(Enzodase)和阿尔特制药公司(Ortho Pharmaceuticals)(Diffusin)。可用于组合物中的透明质酸酶的非限制性实例是人透明质酸酶-1(SEQ IDNO:1)、人透明质酸酶-2(SEQ ID NO:2)、人透明质酸酶-3(SEQ ID NO:3)、人透明质酸酶-4(SEQ ID NO:4)和人PH20(SEQ ID NO:5)。
弹性蛋白酶(EC 3.4.21.36)是称为“丝氨酸蛋白酶”的一组酶的成员,其特征在于酶活性位点中丝氨酸残基的反应性。弹性蛋白酶分解弹性蛋白(弹性纤维的特定蛋白),并消化其他蛋白,例如纤维蛋白、血红蛋白和白蛋白。已经鉴定了三种结构相关的弹性蛋白酶类型,称为弹性蛋白酶I、II和III(或蛋白酶E),哺乳动物的外分泌胰腺分泌了几种同工型。已经证实弹性蛋白酶存在于大多数哺乳动物(包括人、猴、猫、兔等)的胰腺中。本文公开的弹性蛋白酶可以源自任何来源,并且可以通过基因工程技术产生。可以使用的弹性蛋白酶的非限制性实例是人弹性蛋白酶I(SEQ ID NO:6)、人弹性蛋白酶II和人弹性蛋白酶III。
在一些实施例中,透明质酸酶、弹性蛋白酶或其组合在组合物中的有效量为1.0个单位、2个单位、3个单位、4个单位、5个单位、6个单位、7个单位、8个单位、9个单位、10个单位、11个单位、12个单位、13个单位、14个单位、15个单位、16个单位、17个单位、18个单位、19个单位、20个单位、25个单位、30个单位、35个单位、40个单位、45个单位、50个单位、100个单位、125个单位、150个单位、175个单位、200个单位、225个单位、250个单位、275个单位、300个单位、325个单位、350个单位、375个单位、400个单位、425个单位、450个单位、475个单位、500个单位、750个单位、1000个单位、1500个单位、2000个单位、以及介于这些值中的任何两个值之间的任何单个量或任何范围。
在实施例中,组合物中的透明质酸酶、弹性蛋白酶或其组合的有效量选自约0.1wt.%至约25wt.%、约0.1wt.%至约20wt.%、约0.1wt.%至约15wt.%、约0.1wt.%至约10wt.%、约0.1wt.%至约8wt.%、约0.1wt.%至约5wt.%、约0.1wt.%至约4wt.%、约0.1wt.%至约3wt.%、约0.1wt.%至约3wt.%、或约0.1wt.%至约1wt.%。具体实例包括约0.1wt.%、约0.5wt.%、约1wt.%、约2wt.%、约5wt.%、约10wt.%、约25wt.%、以及介于这些值中的任何两个值之间的任何单个量或任何范围。在实施例中,相对于组合物的总量,本文公开的重量百分比可以是重量/重量百分比或重量/体积百分比。
在实施例中,组合物中透明质酸酶、弹性蛋白酶或其组合的有效量选自约1微克至约100毫克/mL组合物、约1微克至约10毫克/mL组合物、约1微克至约5毫克/mL组合物、约1微克至约1毫克/mL组合物、或约1微克至约100微克/mL组合物。
活性剂
各种实施例的组合物可包括几乎任何活性剂,包括用于局部(topical)或局部(local)递送的药剂。活性剂的非限制性实例包括生物制剂、治疗性肽、仿生肽、小分子和大分子镇痛剂、抗真菌剂、抗细菌剂、麻醉剂、蛋白质、前列腺素、酶抑制剂、类固醇、小分子药物、大分子药物、生物制剂、抗体、嵌合抗体、抗体片段、诊断性抗体、抗原、肽、佐剂、抗氧化剂、化妆品成分、治疗性细胞、诊断剂、放射性示踪剂、造影剂、神经毒素、感觉改变剂等及其组合。
实施例所涵盖的生物制剂、治疗性肽和仿生肽包括但不限于肉毒杆菌毒素及其嵌合体或衍生物、抗体、抗体片段、抗体的衍生物、Rejuline、CG-Purilux、CG-Dermaheal、CGKeramin2、Prohairin-β4、CG-TGP2、CG-EDP3、CG-IDP等及其组合。
镇痛剂、抗真菌剂、抗细菌剂和麻醉剂以及类固醇的非限制性实例包括加巴喷丁、普瑞巴林、米诺环素、乙酰水杨酸、环孢菌素、他克莫司(FK506)、比马前列素和其他PGE2抑制剂、他达拉非、克林霉素、可的松、米诺地尔、硫酸米诺地尔、烟酰胺、水杨酸甲酯、加巴喷丁、氢化可的松、棕榈酰-KTTKS肽、苯妥英、维生素B12、环苯扎林、阿那曲唑、利多卡因、视黄酸、丙酸视黄酯、米诺环素、硫酸庆大霉素、比马前列素、硫酸米诺地尔、丙酸氯倍他索、抗坏血酸、氨甲环酸、水杨酸(水杨酸钠)、对苯二酚、托萘酯(tolfnaftate)、克霉唑、特比萘芬、异维甲酸、维甲酸、曲酸、强的松、防晒活性物质(如胡莫柳酯、水杨酸辛酯、奥克立林或阿伏苯宗)、氢化可的松、利多卡因、艾塞吉珠单抗-塔兹(ixekizumab taltz)、氨基乙酰丙酸(ALA)、巴瑞替尼、托法替尼、阿达木单抗、香茅油、3(N-丁基-N-乙酰基)氨基丙酸乙酯、沙瑞环素、D3类似物、钙调磷酸酶抑制剂、氮芥、免疫抗原、咪喹莫特、布洛芬、塞来昔布、双氯芬酸、西地那非、环吡酮(cyclopyrox)、沙瑞环素、雌激素、共轭雌激素等及其组合。
在各种实施例中,活性剂可以是以下一种或多种:α-生育酚、β-胡萝卜素、2-巯基苯并噻唑、阿巴卡韦、阿巴西普、阿昔单抗、香蒿素(abrotanum)、苦艾、阿拉伯树胶、阿坎酸、阿卡波糖、醋丁洛尔、乙酰丙嗪马来酸酯、乙酰乙酸(acetagesic)、对乙酰氨基酚、乙酰唑胺、乙酸、醋羟胺酸、乙酰半胱氨酸、乙酰-酪氨酸、酸性氟磷酸盐、阿维A、阿地溴铵(aclidinium)、乌头草药剂(aconite)、乌头类中药(Aconitum Napellus)、顶头孢霉(Acremonium Cephalosporium)、穗花类叶升麻(Actaea Spicata)、阿昔洛韦、阿达木单抗、阿达帕林、腺嘌呤、腺苷、春福寿草(Adonis Vernalis)、肾上腺素、欧洲七叶树(AesculusHip)、愚人欧芹(Aethusa Cynapium)、阿法替尼、Afoxolaner、捕蝇蕈(AgaricusMuscarius)、牡荆(Agnus Castus)、腺臭椿(Ailanthus Glandulosus)、阿克洛胺、丙氨酸、阿苯达唑、阿比鲁肽、人白蛋白、沙丁胺醇、阿卡他定、阿氯米松、阿地白介素、阿仑膦酸盐、北美肺筋草(Aletris Farinosa)、苜蓿、阿法沙龙、阿芬太尼、阿夫唑嗪、阿利苏单抗(alirocumab)、阿利吉仑半富马酸盐、阿利维A酸、尿囊素、别嘌呤醇、阿莫曲坦、欧洲桤木(Alnus Glutinosa)、芦荟、阿洛司琼、阿普唑仑、前列地尔、缢缩鸭脚树(AlstoniaConstricta)、细链孢霉(Alternaria Tenuis)、烯丙孕素、铝、金刚烷胺、龙涎香、豚草(Ambrosia Artemisiaefolia)、阿米卡星、阿米洛利、氨基己酸、氨基马尿酸盐、氨基乙酰丙酸、氨基戊酰胺、氨茶碱、氨丙嗪、胺碘酮、阿米替林、氨氯地平、氨、异戊巴比妥、阿莫沙平、阿莫西林、苯丙胺、安福霉素、两性霉素B、氨苄青霉素、安普罗林、亚硝酸戊酯(amylnitrosum)、琉璃繁缕(Anagallis Arvensis)、阿那曲唑、无水、阿尼芬净、地蒽酚、阿朴吗啡、阿可乐定、阿泊拉霉素、阿加曲班、金属银、精氨酸、阿立哌唑、阿莫达非尼、山金车酊(arnica)、砷苯酰胺、砷盐、砷、蒿甲醚、阿替卡因、阿魏、欧洲细辛(Asarum Europaeum)、块茎状马利筋(Asclepias Tuberose)、抗坏血酸、阿塞那平马来酸盐、天冬氨酸、阿司匹林、苯磺酸阿曲库铵、豆状滨藜(Atriplex Lentiformis)、颠茄(Atropa Belladonna)、阿托品、凹凸棒石、出芽短梗霉(Aureobasidium Pullularia Pullulans)、溴化金、碘化金、金属金、氯化金、燕麦(Avena Sativa)、阿维巴坦(avibactam)、阿维拉霉素、avobenzoe、阿伏苯宗、阿昔替尼、阿扎胞苷、阿扎哌隆、硫唑嘌呤、壬二酸、氮卓斯汀、阿奇霉素、氨曲南、杆菌肽、巴氯芬、badiaga、巴柳氮、香脂木、秘鲁香脂、班贝霉素、赝靛(BaptisiaTinctoria)、钡、氧化钡、巴利昔单抗、倍氯米松、贝拉西普、贝那普利、苄氟噻嗪、bentoquatam、苄烷铵、苯佐卡因、苯佐那酯、二苯甲酮、过氧化苯甲酰、苄非他明、苯托品、苯甲醇、贝拉克坦(beractant)、β-胡萝卜素、β-氨基丙腈、倍他米松、倍他洛尔、氨甲酰甲胆碱、蓓萨罗丁、贝洛托舒单抗(bezlotoxumab)、比卡鲁胺、比西酸锝(bicisate)、比马前列素、生物素、比沙可啶、金属铋、比索洛尔富马酸酯、比伐卢定、博来霉素、波西普韦、勃地酮、硼砂、硼酸、博舒替尼、灰葡萄孢菌(Botrytis Cinerea)、A型肉毒杆菌毒素、牛生长激素(牛蛋氨生长素锌)、溴莫尼定、布林佐胺、布洛鲁单抗(brodalumab)、溴芬酸、溴、溴隐亭、布地奈德、bultabital、布美他尼、布比卡因、丁丙诺啡、丁喹酯、丁螺环酮、仲丁巴比妥、布他卡因、布他比妥、布他米唑、布替萘芬、布托啡诺、丁基、卡巴他赛、卡麦角林、涩谷花叶芋(Caladium Seguinum)、炉甘石、乙酸钙(calcarea acetica)、砷酸钙(calcarea arsenicica)、碳酸钙(CalcareaCarbonica)、腐蚀性钙、钙粉、氟化钙(calcarea fluorica)、碘化钙(calcarea iodata)、氯化钙(calcarea muriatica)、草酸钙、磷酸钙、硅酸钙、硫化钙(calcarea suflruica)、硫酸钙(calcarea sulphurica)、碳酸氢钙(Calceria Carbonica)、磷酸氢钙(CalceriaPhosphorica)、卡泊三醇、卡泊三醇、骨化三醇、钙、calgest、坎苯达唑、樟脑、康纳单抗、坎地沙坦酯、斑蝥素、斑蝥、卡培他滨、卡莫瑞林、辣椒素、辣椒、卡托普利、乙二磺酸卡拉美芬、卡巴胆碱、卡巴多司、卡马西平、尿素、卡比多巴、动物炭、植物炭、石炭酸、碳霉素、碳、碳酸钙(Carbonate Lime)、碳酸钡、碳酸钾、碳酸钠、carboneum、卡铂、卡前列素氨丁三醇、羧甲基纤维素、奶蓟(Carduus Marianus)、卡芬太尼柠檬酸酯、卡立普多、卡硝唑、卡洛芬、葛缕子(Carum Carvi)、卡维地洛、卡藜、酪蛋白、卡泊芬净、欧洲板栗(Castanea Vesca)、海狸香、葳严仙属、苛性钠(causticum)、策桩子(cedron)、头孢克洛、头孢羟氨苄、头孢唑啉、头孢地尼、头孢吡肟、头孢噻肟、头孢替坦、头孢维星(cefovecin)、头孢西丁、头孢泊肟酯、头孢丙烯、头孢洛林酯、头孢他啶、头孢噻呋、头孢曲松、头孢呋辛、塞来昔布、CenchrisContortrix、风箱树(Cephalanthus Occidentalis)、头孢氨苄、头孢匹林B、色瑞替尼、西替利嗪、十六烷基吡啶、西维美林、球毛壳菌(Chaetomium Globosum)、白屈菜(ChelidoniumMajus)、鹅去氧胆酸(chenodiol)、驱虫藜(Chenopodium Anthelminticum)、伞形梅笠草(Chimaphila Umbellata)、中国硫酸(China Sulphuricum)、药用金鸡纳(ChinchonaOfficinalis)、奎宁(chininum)、氯苯达诺、三氯乙醛、氯胺、氯霉素、氯环嗪、利眠宁、洗必泰、氯(chlorine)、氯(chlorinum)、氯丁醇、氯普鲁卡因、氯喹、氯噻嗪、氯二甲苯酚、氯苯甘醚、氯苯那敏、氯丙嗪、氯磺丙脲、氯四环素、氯噻酮、氯唑沙宗、胆钙化醇、胆固醇(cholesterinum)、消胆胺、绒毛膜促性腺激素α、绒毛膜促性腺激素(chorionicgonadotropin)、含铬的、铬、胰凝乳蛋白酶、环吡酮、毒芹(Cicuta Virosa)、西司他丁、西洛他唑、西咪替丁、温带臭虫(Cimex Lectularius)、总状升麻(Cimicifuga Racemosa)、Cina、海瓜叶菊(Cineraria Maritima)、环丙沙星、顺式阿曲库铵、顺铂、加拿大岩蔷薇(CistusCanadensis)、西酞普兰、柠檬酸、柠檬酸镁(citroma magnesium)、枝孢样枝孢霉(Cladosporium Cladosporioides)、克拉屈滨、克拉霉素、克拉维酸、氯马斯汀、直立铁线莲(Clematis Erecta)、克仑特罗、环奎二苯酯(clidinium)、克林霉素、氯碘羟喹、氯倍他索、氯可托龙、氯膦酸盐、氯法拉滨、克罗米芬、氯米帕明、氯硝西泮、可乐定、氯吡格雷、氯羟吡啶、氯前列醇、氯卓酸盐、氯舒隆、克霉唑、邻氯青霉素、氯氮平、钴胺素、钴、可比司他、可卡因、可待因、秋水仙素、秋水仙碱、考来替泊、粘菌素甲磺酸盐(colistimethate)、檀香基胶原酶(collagenase santyl)、胶态三氧化二铁、胶态硫、药西瓜瓤、复方安息香、康德郎皮(condurango)、毒参属、共轭雌激素、铃兰(Convallaria Majalis)、药用苦配巴香脂(Copaiva Officinalis)、铜、促肾上腺皮质激素、可的松、替可克肽(cosyntropin)、蝇毒磷、尖刺山楂(Cratageus Oxycantha)、甲酚、克唑替尼、番红花(Crocus Sativus)、色甘酸、响尾蛇(Crotalus Horridus)、克罗米通、巴豆(Croton Tiglium)、隐甲藻(Crypthecodinium Cohnii)DHA油、药用荜澄茄(Cubeba Officinalis)、西瓜(CucurbitaCitrullus)、尖音库蚊(Culex Pipiens)、含铜的、铜、不对称弯孢霉(CurvulariaInaequalis)、墨鱼墨、氰钴维生素、欧洲仙客来(Cyclamen Europaeum)、赛克利嗪、环苯扎林、环磷酰胺、环丝氨酸、环孢菌素、赛庚啶、半胱氨酸、阿糖孢苷、畜蜱磷、达比加群、达拉菲尼、达卡巴嗪、达卡他韦、达利珠单抗、Daikon、达伐吡啶(dalfampridine)、达肝素钠、达那唑、达氟沙星、丹曲林、达格列净、氨苯砜、达托霉素、达非那新、达萨布韦、达沙替尼、曼陀罗(Datura Stramonium)、道诺霉素、地西他滨、癸氧喹酯、地拉罗司、去铁胺、地拉韦啶、翠雀属植物(Delphinium Staphisagria)种子、Delsym、地美环素、地拉考昔、地氟烷、地昔帕明、地西卢定、地氯雷他定、地洛瑞林、去氨加压素、去氧孕烯、地奈德、去羟米松、去氧皮质酮、地文拉法辛、地塞米松、右美托咪定、右雷佐生、右旋糖酐、右美沙芬、右旋安非他命、右旋糖、泛影酸盐、安定、二氮嗪、二元的、地布卡因、双氯酚、敌敌畏、重铬酸盐、地克珠利、双氯芬酸、双氯西林、双环维林、地达诺新、乙胺嗪、安非拉酮、二氟拉松、二氟沙星、二氟尼柳、洋地黄(Digitalis Purpurea)、地高辛、二氢麦角胺、二氢链霉素、地尔硫卓、茶苯海明、聚二甲基硅氧烷、二甲基、地诺前列酮、长柔毛薯蓣(Dioscorea Villosa)、Dioscoreinum、苯海拉明、双哌嗪、二丙诺啡、双嘧达莫、地洛他派、丙吡胺、双硫仑、二噻扎宁、双丙戊酸、多巴酚丁胺、多西他赛、Docone、Doconexant、二十二醇、多非利特、狗上皮细胞、毛叶泽兰(DogFennel)、多拉司琼、刺毛扁豆(Dolichos Pruriens)、多潘立酮、多奈哌齐、多巴胺、多拉菌素、多佐胺、多沙普仑、多沙唑嗪、多塞平、度骨化醇、阿霉素、强力霉素、长蠕孢内脐蠕孢(Drechslera Helminthosporium)、屈大麻酚、决奈达隆、氟哌利多、圆叶茅膏菜(DroseraRotundifolia)、屈螺酮、D-硫胺素、度拉糖肽、南蛇藤、度洛西汀、Durezol、度他雄胺、达克罗宁、依莰舒、松果菊(Echinacea Purpurea)、乙膦硫胆碱、益康唑、依法韦仑、艾氟康唑(efinaconazole)、依氟鸟氨酸、依罗霉素、珊瑚色眼镜蛇(Elaps Corallinus)、艾尔巴韦(elbasvir)、依立曲坦、中国榆树(Elm Chinese)、艾曲波帕乙醇胺、乙甲丁酰胺、依美斯汀、艾默德斯(emodepside)、恩格列净、恩曲他滨、依那普利拉、恩夫韦地、依诺肝素、恩诺沙星、恩索利唑、恩他卡朋、恩替卡韦、恩杂鲁胺、麻黄碱、麻黄碱、麻黄碱、黑附球菌(EpicoccumNigrum)、藤地莓(Epigaea Repens)、依匹斯汀、肾上腺素、Epiphegus Virginiana、盐酸表柔比星、依普利酮、依前列醇、依立诺克丁、依普沙坦、依西太尔、依替巴肽、马胸腺细胞免疫球蛋白、问荆(Equisetum Arvense)、木贼(Equisetum Hyemale)、麦角钙化醇、酒石酸麦角胺、加拿大飞蓬(Erigeron Canadensis)、厄他培南、桂竹香(Erysimum Cheiri)、红霉素、艾司西酞普兰、Esium、艾司洛尔、埃索美拉唑、艾司唑仑、酯化的、雌二醇、雌激素、雌酮、雌酮硫酸酯哌嗪(estropipate)、右旋佐匹克隆、依他尼酸、乙胺丁醇、炔雌醇、乙硫异烟胺、乙氧酰胺苯甲酯、乙琥胺、乙醇、乙基异丁嗪、炔诺醇、依替膦酸盐、依托度酸、依托咪酯、依托孕烯、依托泊苷、埃托啡、蒲桃(Eugenia Jambosa)、丁香酚、续随子(Euphorbia Lathyris)、依维莫司、依西美坦、艾塞那肽、印度天南星(Arisaema Triphyllum)根提取物、铁线莲状马兜铃(Aristolochia Clematitis)提取物、亚历桑那光腊树(Arizona Ash)提取物、亚利桑那丝杉(Arizona Cypress)提取物、赝靛(Baptisia Tinctoria)根提取物、红珊瑚(CoralliumRubrum)提取物、加拿大铁杉(Abies Canadensis)提取物、相思豆(Abrus Precatorius)种子提取物、腰果(Anacardium Occidentale)提取物、价如树(Anacardium Orientale)提取物、印度防己(Anamirta Cocculus)种子提取物、山道年蒿(Artemisia Cina)开花前顶端提取物、艾蒿(Artemisia Vulgaris)提取物、三叶海芋(Arum Triphyllum)提取物、亚历桑那光腊树(Ash Arizona)提取、美国白蜡树(Ash White)提取物、芦笋提取物、山杨提取物、烟曲霉(Aspergillus Fumigatus)提取物、黑曲霉(Aspergillus Niger)提取物、澳洲松木(Australian Pine Beefwood)提取物、鳄梨提取物、印度苦楝树(Azadirachta Indica)提取物、百喜草(Bahia Grass)提取物、落羽杉(Bald Cypress)提取物、伏牛花提取物、大麦食品提取物、杨海属月桂果实(Bayberry Wax Myrtle)提取物、蜂毒提取物、山毛榉提取物、牛肉提取物、颠茄叶提取物、雏菊(Bellis)提取物、Berberis Aquarius提取物、冬青叶小檗(Berberis Aquifolium)提取物、欧洲小檗(Berberis Vulg)提取物、欧洲小檗(BerberisVulgaris)提取物、百慕大草提取物、桦木(Betula Alba)提取物、黑桦提取物、红河桦提取物、白桦提取物、苦瓜提取物、黑升麻提取物、石墨提取物、刺槐(Black Locust)提取物、黑胡椒提取物、黑胡桃花粉提取物、黑柳提取物、美国蜚蠊(Blatta Americana)提取物、东方蜚蠊(Blatta Orientalis)提取物、德国小蠊(Blattella Germanica)提取物、一年生早熟禾提取物、梣叶枫(Box Elder Ash Leaf Maple)提取物、巴西胡桃提取物、花椰菜提取物、雀麦草提取物、泻根提取物、荞麦提取物、巨蝮蛇毒提取物、毛茛提取物、甘蓝提取物、夜花仙人掌(Cactus Grandiflorus)提取物、硫酸镉提取物、咖啡因提取物、鲑鱼降钙素提取物、金盏花提取物、加利福尼亚州海岸槲树(Live Oak Coast)提取物、加利福尼亚州胡椒树提取物、加利福尼亚州黑胡桃花粉提取物、甘汞提取物、牛角瓜(Calotropis Gigantea)提取物、白色念珠菌(Candida Albicans)提取物、近平滑假丝酵母(Candida Parapsilosis)提取物、哈密瓜提取物、苋(carelessweed)提取物、胡萝卜提取物、Castor Birch提取物、Castor Equi提取物、蓖麻油提取物、猫毛提取物、猫毛皮提取物、牛上皮提取物、美洲茶(Ceanothus Americanus)提取物、硬叶榆(Cedar Elm)提取物、山雪松(Cedar Mountain)提取物、红雪松提取物、芹菜提取物、甘菊植物提取物、牡荆树提取物、樱桃提取物、鸡肉提取物、榔榆(Chinese Elm)提取物、弗吉尼亚州流苏树(Chionanthus Virginica)提取物、金鸡纳树提取物、肉桂提取物、药西瓜果实提取物、蛤蜊提取物、石松(Club Moss)提取物、煤焦油提取物、仙人掌木防己(Cocculus Cacti)提取物、印度防己提取物、苍耳提取物、可可豆全豆巧克力提取物、可可脂提取物、椰子提取物、鳕鱼提取物、咖啡提取物、二蕊紫苏(Collinsonia Canadensis)提取物、二蕊紫苏根提取物、胶态燕麦片提取物、紫草科植物提取物、紫草科植物根提取物、常见艾蒿提取物、常见蒿属植物提取物、常见一年生苦艾提取物、毒茴(Conium Maculatum)提取物、珊瑚蛇(Coral Snake/Micrurus Fulvius)免疫球蛋白抗蛇毒素(马)提取物、玉米提取物、棉短绒提取物、棉籽提取物、常见东部杨木提取物、弗里蒙特三角叶杨(Cottonwood Fremont)提取物、常见西部杨木提取物、螃蟹提取物、荚迷皮提取物、黄瓜提取物、墨鱼提取物、亚利桑那丝杉提取物、落羽杉(Cypress Bald)提取物、雏菊提取物、蒲公英提取物、印度瑞香提取物、欧亚瑞香(Daphne Mezereum)皮提取物、颠茄(Deadly Nightshade)提取物、小酸模(Dock Sour Sheep Sorrel)提取物、皱叶酸模(DockYellow)提取物、常见东部杨木提取物、狭叶紫锥菊(Echinacea Angustafolia)提取物、狭叶紫锥菊(Echinacea Angustifolia)提取物、蛋白提取物、蛋黄提取物、美国榆树提取物、硬叶榆提取物、埃罗妥珠单抗(elotuzumab)提取物、英国车前草提取物、英国胡桃提取物、英国胡桃花粉提取物、桉树提取物、桉树油提取物、贯叶泽兰(Eupatorium Perfoliatum)提取物、贯叶泽兰花梢提取物、紫苞泽兰(Eupatorium Purpureum)提取物、小米草(Euphrasia)提取物、欧洲接骨木(European Elder)提取物、假豚草(False Ragweed Bur)提取物、比目鱼提取物、芳香漆树提取物、美国白蜡树提取物、弗里蒙特三角叶杨提取物、紫梢花(Freshwater Sponge)提取物、雏菊、墨角藻(Fucus Vesiculosus)提取物、金英树(Galphimia Glauca)花梢提取物、芸香(Garden Rue)提取物、大蒜提取物、金钩吻(Gelsemium Sempervirens)提取物、金钩吻根提取物、斑点老鹳草(Geranium Maculatum)提取物、德国小蠊(German Cockroach)提取物、姜提取物、银杏(Ginkgo Biloba)提取物、山羊奶提取物、秋麒麟草提取物、北美黄莲提取物、续随子(Gopher plant)提取物、葡萄柚提取物、石墨提取物、生咖啡提取物、英国嫩豌豆提取物、几内亚猪上皮细胞提取物、朴树提取物、榛子花粉提取物、念珠蜥蜴(Heloderma Horridum)毒液提取物、谷他血红蛋白(Hemoglobin Glutamer)-200(牛)提取物、蜜蜂提取物、蜜露提取物、啤酒花提取物、七叶树(Horse Chestnut)提取物、马上皮细胞提取物、马尾提取物、印度海扇提取物、吐根提取物、吐根根提取物、弗吉尼亚栎(virginia Live Oak)提取物、德国鸢尾(Iris Germanica)提取物、意大利黑麦草提取物、药喇叭提取物、约翰逊草提取物、核桃(Juglans Regia)提取物、西部刺柏(Juniper Western)提取物、杜松子(Juniperus Communis)提取物、多叶叉子圆柏嫩枝(Juniperus Sabina Leafy Twig)提取物、北美圆柏(Juniperus Virginiana)提取物、南美巨蝮(Lachesis Muta)提取物、羔羊提取物、利马豆提取物、龙虾提取物、黑刺槐非树干提取物、散黑穗病小麦提取物、广玉兰(Magnolia Grandiflora)提取物、红枫提取物、枫糖提取物、贾如树果(Marking Nut)提取物、刺果沼生接骨木(Marshelder Burweed)提取物、粗糙沼生接骨木(Marshelder Rough)提取物、洋甘菊(Matricaria Recutita)提取物、标准化牛尾草(Meadow Fescue Grass)提取物、白千层属花粉提取物、药用草木犀提取物、药用蜜蜂花提取物、土荆芥(Mexican Tea)提取物、镁乳(Milk of Magnesia)提取物、水飞蓟(Milk Thistle)提取物、牛全乳提取物、山金车(Mountain Arnica)提取物、山雪松提取物、山地烟草提取物、小鼠上皮细胞提取物、小鼠上皮细胞提取物、卷枝毛霉(MuscorCircinelloides F.Lusitanicus)提取物、密丛毛霉(Mucor Plumbeus)提取物、总状毛霉(Mucor Racemosus)提取物、常见艾蒿提取物、红桑提取物、白桑提取物、芥菜籽提取物、加利福尼亚州海岸槲树提取物、红橡提取物、弗吉尼亚州槲树提取物、白橡提取物、燕麦粒提取物、燕麦秸秆提取物、野生燕麦花粉提取物、燕麦片提取物、燕麦秸秆提取物、芥菜籽油提取物、老香脂(Old Balsam)提取物、夹竹桃提取物、橄榄提取物、橄榄花粉提取物、洋葱提取物、橙子提取物、野茅提取物、鸢尾草根提取物、牡蛎提取物、棕榈皇后可可棕榈提取物、斑蟊(Spanish Fly)提取物、欧芹提取物、西番莲(Passiflora Incarnata)提取物、西番莲顶部提取物、西番莲(Passion Flower)提取物、桃子提取物、花生提取物、梨提取物、美洲山核桃提取物、美洲山核桃花粉提取物、果胶提取物、加利福尼亚州胡椒树提取物、美洲大蠊提取物、黑云杉(Picea Mariana)树脂提取物、粗糙红根苋提取物、多刺苋提取物、澳洲松木提取物、白松提取物、黄松提取物、菠萝提取物、黑白斑豆(Pinto Bean Kidney Bean)提取物、糖松(Pinus Lambertiana)提取物、樟子松(Pinus Sylvestris)提取物、开心果提取物、大车前草(Plantago Major)提取物、车前种子提取物、英国车前草提取物、李子提取物、毒芹提取物、毒常春藤提取物、毒坚果提取物、毒橡树提取物、药用pongia骨架提取物、白杨提取物、猪肉提取物、金盏花(Pot Marigold)提取物、草原鼠尾草提取物、车前草提取物、纯燧石提取物、紫锥花提取物、匍匐冰草提取物、破斧树提取物、皇后棕榈可可棕榈提取物、腺栎(Quercus Glandium)醑剂提取物、兔提取物、兔上皮细胞提取物、假豚草提取物、短豚草提取物、细长豚草提取物、南部豚草提取物、高大豚草提取物、西部豚草提取物、腐臭牛肉提取物、球根毛茛(Ranunculus Bulbosus)提取物、生鸦片胶提取物、红雪松提取物、红枫提取物、红桑提取物、红橡树提取物、红洋葱提取物、红上繁草提取物、欧鼠李(RhamnusFrangula)提取物、牛皮杜鹃(Rhododendron Aureum)叶提取物、牛皮杜鹃(RhododendronChrysanthum)提取物、深红酵母(Rhodotorula Rubra)提取物、大黄提取物、红色河桦提取物、刺槐(Robinia Pseudoacacia)提取物、粗糙沼生接骨木提取物、粗糙苋提取物、粗糙苋红根提取物、酸模提取物、俄罗斯蓟提取物、芸香提取物、黑麦提取物、黑麦草提取物、意大利黑麦草提取物、草原鼠尾草提取物、常见蒿属植物提取物、鲑鱼提取物、盐草提取物、药用鼠尾草(Salvia Officinalis)提取物、接骨木花提取物、加拿大血根草(SanguinariaCanadensis)提取物、药用肥皂草(Saponaria Officinalis)根提取物、沙巴草(Schoenocaulon Officinale)种子提取物、千里光属提取物、决明属提取物、乌贼提取物、血清促性腺激素提取物、芝麻籽提取物、糙皮山核桃(Shagbark Hickory)提取物、短豚草花粉过敏原提取物、虾提取物、细长豚草提取物、茄属植物提取物、毛果一枝黄花(SolidagoVirgaurea)提取物、毛果一枝黄花花梢提取物、酸模(Sour Dock Sheep Sorrel)提取物、南部豚草提取物、大豆提取物、大豆油提取物、菠菜提取物、多刺苋提取物、药用海绵骨架提取物、南瓜提取物、圣依纳爵豆(St Ignatius Bean)提取物、圣约翰草(St Johns Wort)提取物、茄匍柄霉(Stemphylium Solani)提取物、大荨麻(Stinging Nettle)提取物、草莓提取物、四季豆(String Bean Green Bean)提取物、吕宋果(Strychnos Ignatii)种子提取物、马钱子(Strychnos Nux-Vomica)种子提取物、糖枫提取物、甜玉米提取物、甘薯提取物、标准化黄花草(Sweet Vernal Grass)提取物、枫香树提取物、枫香树非树干提取物、美国梧桐提取物、聚合草提取物、古巴狼蛛(Tarentula Cubensis)提取物、西班牙狼蛛(TarentulaHispana)提取物、金钟柏OCC提取物、烟叶提取物、番茄提取物、金枪鱼提取物、火鸡肉提取物、松脂提取物、松节油提取物、熊果(Uva Ursi)提取物、缬草属植物提取物、香草提取物、蔬菜木炭提取物、绒毛草(Velvet Grass)提取物、白藜芦(Veratrum Album)提取物、白藜芦根提取物、绿藜芦提取物、毛蕊花(Verbascum Thapsus)提取物、多穗马鞭草(VerbenaHastata)提取物、欧洲荚蒾(Viburnum Opulus)提取物、欧洲荚蒾根提取物、香堇菜(ViolaOdorata)提取物、三色堇(Viola Tricolor)提取物、黑胡桃花粉提取物、加利福尼亚州黑胡桃花粉提取物、英国胡桃花粉提取物、水麻提取物、西瓜提取物、西部杨木提取物、西部杜松提取物、西部豚草提取物、小麦花粉提取物、黑穗小麦提取物、白桤木提取物、白蜡树提取物、白桦提取物、白雪松提取物、白桑提取物、白橡树提取物、白色氧化砷提取物、白凡士林提取物、白凡士林矿物油提取物、白松提取物、白杨提取物、白马铃薯提取物、白无核葡萄提取物、全山金车植物提取物、全蛋提取物、全麦小麦籽粒提取物、野生啤酒花提取物、野生薰衣草提取物、野生燕麦花粉提取物、黑柳提取物、银莲花(Wind Flower)提取物、金缕梅提取物、木杂酚油提取物、白英(Woody Nightshade)提取物、土荆芥提取物、一年生常见苦艾提取物、Wyethia Helenioides提取物、Wyethia Helenioides根提取物、酿酒酵母(YeastSaccharomyces Cerevisiae)提取物、皱叶酸模(Yellow Dock)提取物、常绿钩吻(YellowJasmine)提取物、黄松提取物、Protortonia Cacti提取物、依佐加滨、荞麦、泛昔洛韦、法莫替丁、氨磺磷、非班太尔、牛胆汁(Fel Tauri)、非尔氨酯、非洛地平、芬苯达唑、非诺贝特、非诺贝酸、非诺多泮、非诺洛芬、芬前列林、芬太尼、三价铁(Ferric)、富马铁氰化物、火蚁、富马酸亚铁、鱼子、非索罗定、非索非那定、纤维蛋白原、榕属菩提树、Filix Mas、非那雄胺、芬戈莫德、非罗考昔、黄酮、氟卡尼、佛罗贝塔普(Florbetapir)、氟苯尼考、氟康唑、氟胞嘧啶、氟达拉滨、氟代脱氧葡萄糖、氟氢可的松、氟马西尼、氟米松、氟尼缩松、氟尼辛、氟轻松、荧光素、多氟多、氟米龙、氟尿嘧啶、氟西汀、氟甲睾酮、氟奋乃静、氟前列醇、氟拉尔拉纳(Fluralaner)、氟氢缩松、氟西泮、氟比洛芬、氟他胺、氟替卡松、氟伐他汀、氟伏沙明、茴香、叶酸、卵泡刺激素、甲吡唑、甲醛、福尔马林、甲酸、福米卡鲁法、福莫特罗、福沙吡坦二甲葡胺(Fosaprepitant Dimeglumine)、膦甲酸、磷霉素氨丁三醇、福辛普利、磷苯妥英、夫罗曲坦、氟维司群、呋喃唑酮、呋喃苯胺酸、镰刀菌、加巴喷丁、钆贝酸盐、钆双胺、钆特醇、钆弗塞胺、加兰他敏、雪花莲、没食子酸(Gallicum Acidum)、镓、藤黄(Gambogia)、加米霉素、更昔洛韦、加尼瑞克、加替沙星、愈创甘油醚(Gauifenesin)、平铺白珠树、吉非替尼、明胶、吉西他滨、吉非罗齐、庆大霉素、Gentiana Quinqueflora、格拉默、葡庚糖酐铁、格列美脲、格列吡嗪、格洛诺木、胰高血糖素、葡萄糖酸内酯、谷氨酸、谷胱甘肽、格列本脲、甘油、甘氨酸、格隆溴铵、甘草(Glycyrrhiza Glabra)、鼠曲草、金印草根、戈那瑞林、醋酸戈那瑞林、促性腺激素释放因子–白喉类毒素缀合物、戈舍瑞林、草棉、短杆菌肽、格拉司琼、格拉皮普(Grapiprant)、水八角、格拉佐普维(Grazoprevir)、灰黄霉素、米甘菊(Guaco)、格利西力、愈创木酚、愈疮木、愈创甘油醚、格利西力、胍法辛、嗜血杆菌b共轭疫苗(脑膜炎球菌蛋白结合物)、Hahnemanns Causticum、哈西奈德、卤倍他索、常山酮、氟哌啶醇、氟烷、哈洛克酮、金缕梅、Hedeoma Pulegioides、Hekla Lava、向日葵、氦氧混合气、氦、Helleborus Foetidus、黑嚏根草、蠕虫毛霉(Helminthmucor)、Helonias Dioica、血红素铁多肽、菪、肝、肝素、七水、海他西林、羟乙基淀粉、六氯酚、黑克斯胺(Hexaminolevulinate)、组胺、组氨酸、后马托品、胡莫柳、人体胰岛素、人乳头瘤病毒9价疫苗、人乳头瘤病毒四价(6、11、16、18型)疫苗、人重组、人Rho(D)免疫球蛋白、啤酒花(Humulus Lupulus)、透明质酸、透明质酸酶、海科地松(Hydorcortisone)、肼苯哒嗪、乔木绣球、金印草、盐酸、氢氯噻嗪、氢可酮、氢化可的松、积雪草(Hydrocotyle Asiatica)、氢氟酸、氢、氢化棕榈仁油、氢吗啡酮、对苯二酚、水合、羟钴胺素、羟氯喹、羟乙基、羟基脲、盐酸羟、潮霉素B、莨菪碱、尼日尔天仙子、金丝桃、羟丙甲纤维素、伊班膦酸、屈曲花(Iberis amara)、布洛芬、伊布利特、鱼石脂、艾考糊精、二十碳五烯酸、伊达比星、达鲁苏麻(Idarucizumab)、异环磷酰胺、吕宋果(Ignatia Amara)、伊格纳修斯豆、鸢尾花(Iiris Versicolor)、伊潘立酮、伊马替尼、吡虫啉、咪苯脲、亚胺培南、丙咪嗪、咪喹莫特、免疫球蛋白(人)、不纯的钙、肉毒杆菌毒素、茚达特罗、吲达帕胺、靛青、茚地那韦、铟、吲哚美辛、英夫利昔单抗、流感病毒疫苗、因弗拉姆(Influenzinum)、巨大戟醇、胰岛素、干扰素、碘化酊、碘化酪蛋白、碘(Iodine)、碘苯二胺葡甲胺(IodipamideMeglumine)、碘(Iodium)、碘克沙醇、碘氯羟喹、碘海醇、碘帕醇、碘他、碘佛醇、吐根、易普利姆玛、异丙托溴铵、厄贝沙坦、铱、伊立替康、邦特耐克丝、鸢尾花、铁、艾萨康佐姆(Isavuconazonium)、伊索地姆(Isodium)、异氟、异氟醚、异亮氨酸、美汀、异烟肼、异丙胺、异丙醇、异丙肾上腺素、硝酸异山梨、异维A酸、伊拉地平、伊曲康唑、伊维菌素、伊沙匹隆、碘克珠单抗(Ixekizumab)、巴西蓝花楹、Jacobaea Maritima、鸭嘴花(Justicia Adhatoda)、亚砷酸钾、砷酸钾、Kali Bechromate、Kali Bechromate Karaya Gum Bassora、标准化KaliBechromate Kentucky Bluegrass(六月)、Kali Bechromate Kochia Firebush、KaliBechromate Krameria Lappacea根、Bechromate Lemon、Kali Bechromate LeopardsBane、Kali Bechromate Lettuce、Kali Bichromicum、Kali Bromatum、卡利(Kali)碳酸盐、卡利(Kali)炭、Kali Iodatum、Kali Muriaticum、Kali Muriaticum、矽剂、硝酸卡利、磷酸卡利、磷酸卡利、Kali Sulphuricum、磷酸卡利、磷酸氧化酶、Natrum Phosphoricum、KaliSulph、Kali Sulphuricum、山月桂、硫酸卡那霉素、木棉花、氯胺酮、氯胺酮、酮康唑、酮洛芬、酮咯酸、酮替芬、酮替芬、木馏油、拉贝洛尔、犬乳、脱脂乳(Lac Defloratum)、LacFelinum、牛乳(Lac Vaccinum)、Lachnanthes Tinctoria、拉科胺、乳酸(Lactic Acid或Lacticum Acidum)、毒莴苣、乳果糖、莱特洛霉素、短柄野芝麻、拉米夫定、拉莫三嗪、羊毛脂、兰瑞肽、兰索拉唑、拉帕替尼、硅氟化钙、牛蒡、拉沙里菌素、拉坦前列素、山黧豆、黑寡妇蜘蛛、月桂酸、桂樱属植物、泻药、L-半胱氨酸、铅、卵磷脂、杜、杜香、杜香枝、来氟米特、浮萍(Lemna Minor)、弗吉尼亚九阶草(Leptandra Virginica)、莱西努拉(Lesinurad)、来曲唑、亮氨酸、亚叶酸钙、亮丙瑞林、左旋沙丁胺醇、左旋咪唑、左乙拉西坦、左旋布诺洛尔、左卡尼汀、左旋多巴、左氧氟沙星、醛氢叶酸、左亚叶酸、左米那普仑、可巴君、左炔诺孕酮、左啡诺、左甲状腺素、果糖、利多卡因、百合(Lilium Tigrinum)、利那洛肽、利格列汀、林可霉素、林丹、利奈唑胺、亚麻酸、三碘甲状腺氨酸、利拉鲁肽、赖诺普利、锂、利西拉、老龙皮提取物、北美山梗菜、洛度沙胺氨丁三醇、洛哌丁胺、洛匹那韦、氯雷他定、劳拉西泮、氯沙坦、氯替泼诺、洛伐他汀、洛沙平、鲁比前列酮、虱螨脲、具盖丝瓜、卢格尔斯(Lugols)、鲁利康、本芴醇、鲁前列醇、叶黄素、石松、弗吉尼亚地笋(Lycopus Virginicus)、赖氨酸、斑蝥(LyttaVesicatoria)、粗晶质、马杜霉素铵、Mag Phos、镁、马拉硫磷、锰(Manganese或Manganum)、甘露醇、盐酸马普替林(Maprotiline Hydrochloride)、马拉韦罗、麻保沙星、马罗匹坦(Maropitant)、马克齐德(Maxzide)、甲苯咪唑、甲溴菲宁、加明、美卡舍明、氮芥、氯苯甲嗪、甲氯灭酸、美托咪定、甲羟孕酮、甲芬那酸、甲氟喹、甲地孕酮、美拉索明、退黑激素、醋酸美拉索明、美洛昔康、马法兰、美金刚、欧薄荷、薄荷醇、睡菜、美喷酯、哌替啶、MephitisMephitica、甲哌卡因H.、美泊利单抗、眠尔通、美拉地酯、巯嘌呤、Mercurius Corrosivus、氯化亚汞(Mercurius Dulcis)、Mercurius Iodatus Flavus、Mercurius Iodatus Ruber、稀释水银(Mercurius Solubilis)、汞、汞、美罗培南、巯替肽、氨水杨酸、美司钠、梅斯基特、美他沙酮、二甲双胍、美沙酮、甲基苯丙胺、醋甲唑胺、乌洛托品、甲巯咪唑、蛋氨酸、美索巴莫、甲氨蝶呤、甲氧沙林、甲氧基聚乙二醇-β红细胞生成素、甲基东莨菪碱、甲琥、甲氯噻嗪、水杨酸甲酯、甲基多巴、亚甲蓝、马来酸甲麦角新碱(Methylergonovine Maleate)、哌甲酯、甲基强的松、甲泼尼龙、水杨酸甲酯、甲睾酮、胃复安、托拉、美托洛尔、美托舍酯、甲硝唑、美西律、洋种瑞香、米勃酮、米卡芬、达克宁、咪达唑仑、米格列醇、美格鲁特、米尔贝霉素肟、蓍、米那普仑、米力农、米诺环素、米诺地尔、米拉贝隆、米氮平、米索前列醇、丝裂霉素、米托坦、米托蒽醌、美维松、莫达非尼、莫尔普利、钼、糠酸莫米他松、莫能菌素、一碱的(Monobasic)、一水合物、孟鲁司特、莫仑、吗啡、鱼肝油、麝、莫昔克丁、莫西沙星、莫匹罗星、紫螺(Murex Purpurea)、氯酸(Muriaticum Acidum)、麦科菲你(Mycophenilic)、麦盖尔(Mygale)、蜡杨梅树皮、Myristica Sebifer、肉豆蔻、大麻隆、萘丁美酮(Nabumentone)、纳多洛尔、那法瑞林、奈夫西林、萘替芬、Naja Tripudians、纳布啡、烯丙吗啡、纳洛格(Naloxegol)、纳洛酮、纳曲酮、诺龙、萘甲唑啉、萘、萘普生、甲基盐霉素、那拉曲坦、肉豆蔻磷(Phos Nutmeg)、纳他霉素、那格列奈、钠、奈必洛尔、奈昔、奈多罗米、奈法唑酮、奈拉滨、新霉素、新斯的明、奈帕芬、奈喹酯、脉孢菌(Neurospora Intermedia)、中性氟化钠、奈韦拉平、烟酸、尼卡巴嗪、尼卡地平、镍、尼古丁、硝苯地平、黑孢、尼罗替尼、尼鲁米特、尼莫地平、尼达尼布、尼索地平、烯啶虫胺、硝酸、呋喃妥因、呋喃西林、氮、硝酸甘油、硝米特、笑气、纳武单抗、尼扎替丁、甲基孕酮、去甲肾上腺素、炔诺酮、诺孕、诺孕美特、甲基炔诺酮、去甲替林、新生霉素、肉豆蔻(Nux Moschata)、马钱子、制霉菌素、圣罗勒、奥地诺兹(Ocitnoxate)、奥斯赛莱(Ocitsalate)、奥克拉替尼(Oclacitinib)、奥西诺酯、水杨酸辛酯、辛苯酮(Octobenzone)、奥克立林、奥曲肽、花色水芹、奥法木单抗、氧氟沙星、奥氮平、奥拉帕尼、奥拉他单抗(Olaratumab)、油酸钠、Olmesartan Medoxmil、奥达特罗、奥洛他定、奥沙拉嗪、奥比沙韦(Ombitasvir)、奥美拉唑、肉毒杆菌素(Onabotulinumtoxina)、昂丹司琼、Onosmodium Virginianum、奥菲那(Oophorinum)、鸦片、仙人掌(Opuntia Vulgaris)、奥比沙星、奥古蛋白、奥利司他、奥美普林、柠檬酸邻甲苯海明、磷酸奥塞米韦、奥希替尼、锇金属、Ova Tosta、绵羊地高辛免疫Fab、苯唑、草酸(Oxalicum Acidum)、奥沙利铂、氧雄龙、奥沙普秦、奥沙西泮、奥卡西平、奥克地诺兹(Oxctinoxate)、奥苯达唑、奥昔、铝的氧化物、氧苯酮、奥昔布宁、羟考酮、氧、羟甲唑啉、羟吗啡酮、羟基喹啉、土霉素、催产素、紫杉醇、二甲胺基苯甲酸戊酯O、荷兰芍药(Paeonia Officinalis)、帕博西尼、帕潘立酮、钯金属、帕马溴、帕米膦酸二钠、胰、泮库溴铵、帕比司他、泮托拉唑、泛酸、罂粟碱、石蜡、帕雷拉布拉瓦、帕立骨化醇、四叶重楼、帕替帕韦(Paritaprevir)、帕罗西汀、帕瑞肽、帕唑帕尼、PEG-3350、培门冬酶、培波格拉斯地姆(Pegbovigrastim)、聚乙二醇干扰素Alfa-2a、聚乙二醇干扰素Alfa2b、培维索孟、派姆单抗、培美曲塞、喷昔洛韦、青霉胺、青霉素G、青霉素V、产黄青霉菌、光孢青霉、罗克福尔青霉菌、五价、喷他佐辛、戊巴比妥、喷司他丁、己酮可可碱、全氟丙烷、甲磺酸培高利特(Pergolide Mesylate)、特丁胺培哚普利Erbumine(PerindoprilErbumine)、氯菊酯、奋乃静、凡士林、石油、欧芹、Phellandrium Aquaticum、那吡啶、苯甲曲秦、苯乙肼、马来酸非利拉明、苯巴比妥、苯酚、吩噻嗪、酚苄明、苯基扎吡啶(Phenozapyridine)、芬特明、酚妥拉明、苯肾上腺素(Phenykephrine)、水杨酸苯酯、苯丙氨酸、苯基苯并咪唑磺酸、保泰松、去氧肾上腺素、苯丙醇胺、苯托沙敏、苯妥英钠、草茎点霉、磷、亚胺硫磷、铁的磷酸盐、磷酸化碳水化合物、磷酸、磷、酸浆(Physalis Alkekengi)、毒扁豆、Phytolacca Americana Root、垂序商陆、植物甲萘醌、苦味酸、苦味酸(PicricumAcidum)、盐酸毛果芸香碱、毛果芸香属、吡美莫司、匹莫、吲哚洛尔、吡格列酮、哌拉西林、哌嗪、增效、吡利霉素、吡罗昔康、垂体促黄体激素、Pix Liquida、铂、普乐沙福、铅、普达非洛、鬼臼、鬼臼树脂、泊洛沙林、聚乙烯、多粘菌素、聚氧乙烯、猪苓、聚山梨醇酯80、多硫酸化糖胺聚糖、聚乙烯醇、帕托珠利、猪肺磷脂、猪、泊沙康唑、钾、Pothos Foetidus、聚维酮、普拉沙星、氯化派姆、普拉克索、普兰林肽、普拉莫星、普拉格雷、普伐他汀、吡喹酮、哌唑嗪、泼尼卡酯、泼尼松龙、强的松、普瑞巴林、丙胺卡因、伯氨喹、扑痫酮、女贞、丙磺舒、普鲁卡因胺、丙氯拉嗪、黄体激素、氯胍、脯氨酸、普马、异丙嗪、普罗帕酮、丙苯丙嗪(Propiopromazine)、异丙酚、丙氧芬、普萘洛尔、丙烯、丙己、丙基硫氧嘧啶、前列他林、普罗、聚维酮碘、黑刺李(Prunus Spinosa)、伪麻黄碱、短梗霉(Pullularia Pullulans)、白头翁、噻嘧啶、吡嗪酰胺、除虫菊、吡啶斯的明、吡哆醇、拉明、乙胺嘧啶、吡啶硫酮锌、派洛济宁(Pyrogenium)、苦木(Quassia Amara)、喹硫平、喹那普利、奎尼丁、拉巴司他定(Rabacfosadine)、雷贝拉唑、消旋肾上腺素(Racepinephrine)、莱克多巴胺、镭、雷洛昔芬、拉替拉韦、雷米普利、雷莫芦单抗、雷尼替丁、萝卜(Raphanus Sativus)、雷沙吉兰、拉布立酶、Ratanhia、蛇根木、重组、瑞加德松(Regadenoson)、瑞格列奈、利血平、间苯二酚、瑞他莫林、大黄、铑、光滑漆树、稻、利巴韦林、里博西利布(Ribociclib)、核黄素、蓖麻(RicinusCommunis)、利福布丁、利福平、利福喷丁、利鲁唑、肉毒毒素b(Rimabotulinumtoxinb)、金刚乙胺、利美索龙、利塞膦酸钠、利培酮、利托那韦、利斯的明、利扎曲普坦、罗贝考昔、氯苯胍、胃长宁、罗库溴铵、罗氟司特、罗米非定、罗匹尼罗、罗哌卡因、马来酸罗格列酮、瑞舒伐他汀钙、洛克沙胂、风疹、铷、芸香(Rue)、沙巴藜芦、沙巴棕果(Sabal Serrulata)、圆柏、酿酒酵母(Saccharomycescerevisiae)、甘蔗乳酸、萨库比尔(Sacubitril)、水杨酸、水杨酰胺、盐水、盐霉素、黑柳(Salix Nigra)、沙美特罗、双水杨酯、Samarium SM 153Lexidronam、山道年(Santoninum)、甲磺酸沙奎那韦、Sarcolacticum Acidum、沙格司亭、帚枝霉滇榄(SarocladiumStrictum)、萨洛纳(Sarolaner)、紫瓶子草、菝葜、沙格列汀、裂殖壶菌Dha油、东莨菪碱(Scopalamine或Scopolamine)、林生玄参、黄芩(Scutellaria Lateriflora)、麦角(SecaleCornutum)、司可巴比妥、苏金单抗、司拉美、多花斯温漆木(Selan)、司来吉兰、硒、硒代蛋氨酸、生度米星(Semduramicin)、番泻甙、丝氨酸、舍他、舍曲林、碳酸司维拉姆、七氟醚、鲨鱼肝油、西地那非、二氧化硅、硅、银、二甲基硅油、辛伐他汀、黑芥子(Sinapis Nigra)、辛卡利特、西尼坎特肯(Sinecatechins)、西罗莫司、西他列汀、斯卡托勒姆(Skatolum)、钠、红火蚁、生长激素、索尼吉布(Sonidegib)、山梨糖醇、索他洛尔、大观霉素、驱虫草、多杀菌素、安体舒通、Spongia Tosta、氯化亚锡、康力龙、Staphysagria、淀粉、司他夫定、鹅肠草(Stellaria Media)、牛皮叶肺草(Sticta Pulmonaria)、Stigmata Maidis、曼陀罗、链霉素、链脲佐菌素、锶、箭毒羊角拗(Strophanthus Hispidus)、琥珀酰胆碱、硫糖铝、舒芬太尼、舒更葡糖、舒巴坦、康唑、磺溴嘧啶、磺胺醋酰、磺胺氯达嗪、磺胺嘧啶、磺胺二甲氧嘧啶、磺胺乙氧嗪、磺胺甲基嘧啶、磺胺二甲嘧啶、磺胺甲、磺胺甲恶唑、磺胺、磺胺硝苯、磺胺喹恶啉、柳氮磺胺吡啶、磺胺异恶唑、磺粘菌素、硫、舒林酸、硫化物的锑、硫、舒马曲坦、舒马曲坦、琥珀酸、五福花根、苹果酸舒尼替尼、苏沃雷生(suvorexant)、蒲桃(SyzygiumJambolanum)、烟草属(Tabaccum)、烟草属高大豚草(Tabaccum Tall Ragweed Giant)、他克莫司、他达拉非、滑石、他立苷酶α(Taliglucerase Alfa)、柠檬酸他莫昔芬、盐酸坦索罗辛、菊蒿(Tanacetum Vulgare)、单宁酸、塔喷他多(Tapentadol)、蒲公英(TaraxacumOfficinalis)、吐酒石(Tartaremetic)、酒石酸(Tartaricum Acidum)、牛磺酸、塔瓦布尔(Tavaborole)、他扎罗汀、他唑巴坦、他唑巴坦、锝、替比夫定、泰利霉素、碲金属、替米沙坦、替马西泮、替莫唑胺、西罗莫司、富马酸替诺福韦地索普西、替泊沙林、特拉唑嗪、特比萘芬、特布他林、特康唑、松脂(Terebinthina)、特立帕肽、睾酮、破伤风、丁卡因、四环素、四氟硼酸盐、四氢唑林、四、马香科(Teucrium Marum)、铊、亚铊、Thaspium Aureum、茶树(TheaSinensis)、噻苯铵醚、茶碱、球腹、噻菌灵、硫烯丙巴比妥、硫胺素(Thiamin或Thiamine)、硫戊巴比妥、硫喷妥钠、硫利达嗪、硫代氨基胺(Thiosinaminum)、硫链丝菌素、噻替哌、替沃噻吨、遏蓝菜巴斯德氏菌(Thlaspi Bursa-Pastoris)、苏氨酸、凝血酶(人)、麝香草酚、百里香胸腺(Thymus Serpyllum)、甲状腺素(Thyroidinum)、噻加宾、泰妙菌素、替卡格雷、替卡西林、噻氯匹定、替加环素、泰地罗新(Tildipirosin)、噻环乙胺、菩提叶、替米考星、膦酸盐、马来酸噻吗洛尔、安息香酊剂、替硝唑、康唑、硫普罗宁、噻昔达唑、替拉那韦、钛、替扎尼定、Tl 201、妥布霉素、托克拉尼(Toceranib)、生育酚酸、琥珀酸、托法替尼、妥拉磺脲、妥拉苏林、甲苯磺丁脲、托卡朋、托美汀、托萘酯、托特罗定、甲苯、托吡酯、拓扑替康、托瑞米芬、托拉塞米、漆树属毛竹(Toxicodendron Pubescens)叶子、曲马多、曲美替尼、群多普利、传明酸、苯环丙胺、曲伏前列素、曲唑酮、群勃龙、维甲酸、曲安西龙、氨苯蝶啶、三唑仑、三碱的、三氯敌百虫(TricaieTrichlorfon)、三氯甲烷(Trichlormethiazie)、三氯乙酸、皮肤毛癣菌、三氯卡班、三氯生、曲恩汀、三氟拉嗪、三叶草、梯牧草(Pratense)、三叶草、白三叶(Repens)、苯海索、曲洛司坦、异丁嗪、三甲双酮、甲氧苄啶、吡苄明、曲普利啶、三乙醇胺、氨丁三醇、托品酰胺、曲司、胰蛋白酶、色氨酸、托拉菌素、泰乐菌素(Tylosin或Tylvalosin)、酪氨酸、乌克地尼(Umeclidinium)、十一烯酸、铀硝酸、尿素、熊去氧胆酸、欧荨麻(UrticaUrens)、Ustilago Maidis、伐昔洛韦、缬更昔洛韦H、缬氨酸、丙戊酸钠、丙戊酸、缬沙坦、万古霉素、凡德他尼、伐地那非、瓦伦尼克林、加压素、维库溴铵B、维内托克斯(Venetoclax)、文拉法辛、维兰特罗、维拉佐酮、小蔓长春花(Vinca Minor)、长春新碱、长春瑞滨、维吉尼亚、槲寄生专辑、维生素A、维生素B6、维生素C、维生素D、维生素D3、维生素E、沃拉帕沙(Vorapaxar)、伏立康唑、伏立诺他、Wal-Zan、Wal-Zyr、华法林、花椒(XanthoxylumFraxineum)、X射线、甲苯噻嗪、育亨宾(Yohimbine或Yohimbinum)、扎鲁司特、扎来普隆、扎那米韦、扎瓦拉(Zavara)、玉米赤霉醇、齐多夫定、齐留通、齐帕特罗、锌、生姜(ZingiberOfficinale)、齐拉西酮、阿柏西普(Ziv-Aflibercept)、二硝托胺、唑拉西泮、唑来膦酸、佐米曲普坦、唑吡坦、唑尼沙胺。
在一些实施例中,活性剂可以是赖诺普利、阿托伐他汀、左旋甲状腺素、氨氯地平、奥美拉唑、二甲双胍、加巴喷丁、辛伐他汀、阿莫西林、氢氯噻嗪、舍曲林、氯沙坦、阿普唑仑、呋塞米、阿奇霉素、布洛芬、美托洛尔、强的松、曲马多、氟替卡松、孟鲁司特、泮托拉唑、依他普仑、及其组合。
在一些实施例中,活性剂选自镇痛剂、抗真菌剂、抗细菌剂、麻醉剂、抗炎剂、抗酒渣鼻剂、血管收缩剂、抗痤疮剂、抗跛行剂、焕肤剂(skin retexturing agent)和类固醇,包括但不限于类视黄醇(视黄醇、视黄醛、视黄酸、视黄醇丙酸酯)、水杨酸盐(乙酰水杨酸、水杨酸甲酯、水杨酸)、过氧化苯甲酰、米诺环素、盐酸克林霉素、磷酸克林霉素、红霉素、四环素、双氯青霉素、强力霉素、维甲酸、异维甲酸、阿达帕林、加巴喷丁、普瑞巴林、环孢菌素、他克莫司(FK506)、羟甲唑啉、溴莫尼定、四氢唑啉、苯肾上腺素、氯吡格雷、替卡格雷、噻氯匹定、比马前列素和其他PGE2抑制剂、他达拉非、克林霉素、可的松、米诺地尔、米诺地尔硫酸盐、烟酰胺、加巴喷丁、氢化可的松、棕榈酰-KTTKS肽、苯妥英、维生素B12、环苯扎林、阿那曲唑、利多卡因、米诺环素、硫酸庆大霉素、比马前列素、硫酸米诺地尔、丙酸氯倍他索、抗坏血酸、氨甲环酸、水杨酸(水杨酸钠)、对苯二酚、托萘酯、克霉唑、特比萘芬、异维甲酸、维甲酸、曲酸、强的松、防晒活性物质(例如胡莫柳酯、辛水杨酯、奥克立林或阿伏苯宗)、氢化可的松、利多卡因、艾塞吉珠单抗-塔兹、氨基乙酰丙酸(ALA)、色瑞替尼、托法替尼、阿达木单抗、香茅油、3(N-丁基-N-乙酰基)氨基丙酸乙酯、沙瑞环素、D3类似物、钙调磷酸酶抑制剂、氮芥、免疫抗原、咪喹莫特、布洛芬、塞来昔布、双氯芬酸、西地那非、环吡酮、沙雷环素、雌激素、共轭雌激素氢氧化钾、鬼臼脂、斑蝥素、咪喹莫特、硝酸、口服西咪替丁、5-氟尿嘧啶、博来霉素、DNCB、咪喹莫特和三氯乙酸、博来霉素、2,4-二硝基氯苯、氟尿嘧啶、硝酸银、硫酸锌、氧化锌、杆菌肽、金霉素、新霉素、莫匹罗星、多粘菌素B、铜迈星、呋喃唑酮、庆大霉素、林可霉素、头孢菌素、β内酰胺抗生素、盐酸林可霉素、他扎罗汀、维生素A、阿维A、蓓萨罗丁、奥昔布宁、维生素D、维生素C、维生素B、维生素E、硫、糖皮质激素、皮质类固醇、曲安西龙、曲安奈德、倍他米松、倍他米松1 7-戊酸酯、倍他米松二丙酸酯、哈西奈德、醋酸异氟泼尼松、氟米松、氟轻松、莫米松、氟替卡松、丙酸氟替卡松、泼尼松龙、倍氯米松、氢化可的松、环丙孕酮、屈螺酮、雌激素、孕激素、他克莫司、吡美莫司、熊果酸、桦木酸、模绕酮酸(moronic acid)、齐墩果酸、阿昔洛韦、伐昔洛韦、泛昔洛韦、喷昔洛韦、二十二醇、紫苏醇、及其组合。
在一些实施例中,活性剂可以用于兽医用途。这些试剂包括但不限于2-巯基苯并噻唑、马来酸乙酰丙嗪、乙酰唑胺钠、乙酰水杨酸、afoxolaner、阿克洛胺、阿苯达唑、硫酸沙丁胺醇、阿法沙龙、烯丙孕素、硫酸阿米卡星、硫酸氢氨戊酰胺、氨丙嗪延胡索酸酯、双甲脒、氯化铵、三水合阿莫西林、安福霉素、无水氨苄西林、氨苄西林钠、三水合氨苄西林、氨丙啉、硫酸安普霉素、砷酸胺钠、盐酸阿替美唑、阿托品、绿坡缕石、阿维拉霉素、阿扎哌隆、亚甲基双水杨酸杆菌肽、杆菌肽锌、秘鲁香油、班贝霉素、β-氨基丙腈、倍他米松戊酸酯、乙酸倍他米松、二丙酸倍他米松、磷酸钠倍他米松、戊酸倍他米松、次碳酸铋、十一碳烯酸宝丹酮(boldenone undecylenate)、牛生长激素(牛蛋氨生长素锌)、盐酸丁萘脒、布比卡因、丁丙诺啡、丁喹酯、硫酸布他卡因、盐酸丁咪唑(butamisole hydrochloride)、酒石酸布托啡诺、堪苯达唑、卡莫瑞林、乙二磺酸卡拉美芬、卡巴多、碳霉素、二氧化碳、柠檬酸卡分替里(carfentilil citrate)、卡硝唑、卡洛芬、蓖麻油、头孢羟氨苄、头孢维星钠、头孢泊肟酯、头孢噻夫晶体游离酸、盐酸头孢噻夫、头孢噻夫钠、头孢氨苄、头孢匹林苄星青霉素、头孢匹林钠、水合氯醛、氯胺-t三水合物、氯霉素、氯霉素棕榈酸酯、醋酸氯己定、盐酸氯己定、氯丁醇、磷酸氯喹、氯噻嗪、氯苯甘油氨酯、金霉素、硫酸氢氯四环素、金霉素钙络合物、盐酸金霉素、绒毛膜促性腺激素、胰凝乳蛋白酶、柠檬酸、克拉维酸钾、盐酸克伦特罗、盐酸克林霉素、氯膦酸盐、盐酸氯丙咪嗪、氯羟吡啶、氯前列醇钠、氯舒隆、克霉唑、苄星邻氯青霉素、邻氯青霉素钠、粘菌素甲磺酸钠、胶状氧化铁、环烷酸铜、促肾上腺皮质激素、蝇毒磷、甘氨酸铜、环孢霉素、环孢菌素口服液、畜蜱磷(cythioate)、达氟沙星、癸氧喹酯、地拉考昔、醋酸德舍瑞林、去氧皮质酮新戊酸酯、盐酸地托咪啶、地塞米松、地塞米松磷酸钠、地塞米松-21-异烟酸酯、右美托咪啶、盐酸右美托咪啶、右旋糖、泛影葡胺、泛影酸钠、盐酸二丁卡因、菌霉净、敌敌畏、地克珠利、双氯芬酸钠、双氯西林钠一水合物、枸橼酸乙胺嗪、盐酸二氟沙星、硫酸双氢链霉素、二甲亚砜、地诺前列素氨丁三醇、硫酸双哌嗪、盐酸二丙诺啡、dirlotapide、碘二噻宁、多潘立酮、多拉菌素、盐酸多沙普仑、盐酸多西环素、琥珀酸多西拉敏、氟哌利多、依罗霉素、乙甲丁酰胺(embutramid)、emodepside、马来酸依那普利、恩诺沙星、依立诺克丁、依西太尔、红霉素、磷酸红霉素、硫氰酸红霉素、雌二醇、苯甲酸雌二醇、戊酸雌二醇、雌三醇、乙氧苯酯、盐酸乙基异丁嗪、依托度酸、盐酸埃托啡、氨磺磷、苯硫胍、芬苯达唑、芬前列林、芬太尼、枸橼酸芬太尼、倍硫磷、非罗考昔、氟苯尼考、氟米松、醋酸氟米松、氟尼辛葡甲胺、氟轻松、盐酸氟西汀、氟前列醇钠、fluralaner、促卵泡激素、福尔马林、呋喃唑酮、呋塞米、加米霉素、明胶、硫酸庆大霉素、硫酸庆大霉素usp、葡庚糖酐铁、甘氨酸、格隆溴铵、醋酸戈那瑞林、四水合双醋酸戈那瑞林、盐酸戈那瑞林、促性腺激素释放因子-白喉类毒素结合物、戈雷匹仑(grapiprant)、灰黄霉素、愈创甘油醚、氢溴酸常山酮、氟烷、哈洛克酮、氦、谷他血红蛋白-200(牛)、缩酮氨苄青霉素钾、透明质酸钠、盐酸盐、氢氯噻嗪、氢化可的松、醋酸丙酸氢化可的松、醋酸氢化可的松、过氧化氢、潮霉素b、吡虫啉、二丙酸咪多卡、碘化酪蛋白、氯碘羟喹、右旋糖酐铁络合物、醋酸异氟泼尼松、异氟醚、异丙碘铵、伊曲康唑、伊维菌素、硫酸卡那霉素、氯胺酮、盐酸氯胺酮、酮洛芬、来洛霉素丙酸钾、拉沙里菌素、拉沙里菌素钠、左旋咪唑、盐酸左旋咪唑、磷酸左旋咪唑、树脂酸左旋咪唑、左甲状腺素钠、利多卡因、林可霉素、盐酸林可霉素、盐酸林可霉素一水合物、碘塞罗宁钠、虱螨脲、鲁前列醇、马杜霉素铵、硫酸镁、马波沙星、马罗匹坦(maropitant)、甲苯咪唑、甲氯芬那酸、美托咪啶、医用空气、醋酸甲地孕酮、美拉索明二盐酸盐(melarsomine dihydrochloride)、褪黑素、醋酸美仑孕酮、美洛昔康、盐酸马比佛卡因、扁桃酸马比佛卡因、美索巴莫、甲基强的松龙、醋酸甲基强的松龙、盐酸美托舍酯、米勃龙(mibolerone)、硝酸咪康唑、米尔比霉素肟化物、糠酸莫美他松、无水糠酸莫美他松、糠酸莫美他松一水合物、莫能菌素、莫能菌素钠、莫能菌素usp、酒石酸莫仑太、莫昔克丁、莫匹罗星、肉豆蔻基-γ-甲基吡啶鎓氯化物、盐酸丙烯吗啡、盐酸纳曲酮、萘普生、甲基盐霉素、正丁基氯、正丁基东莨菪碱溴化铵(n-butylscopolammoniumbromide)、新霉素、新霉素棕榈酸酯、硫酸新霉素、甲基硫酸新斯的明、奈喹酯nf、尼卡巴嗪、烯啶虫胺、呋喃西林、氮、硝米特、一氧化二氮、诺孕美特、新生霉素、新生霉素钠、制霉菌素、oclacitinib、油酸钠、奥美拉唑、opafp-ghc2 rdna构建体、奥比沙星、奥古蛋白、奥美普林、奥芬达唑、奥苯达唑、氧、土霉素、土霉素(单烷基三甲基铵盐)、土霉素二水合物、盐酸土霉素、催产素、pegbovigrastim、苄星青霉素、青霉素G钾、普鲁卡因青霉素、青霉素V钾、乳酸喷他佐辛、戊巴比妥、戊巴比妥钠、甲磺酸培高利特、吩噻嗪、保泰松、盐酸苯丙醇胺、苯妥英钠、亚胺硫磷、匹莫苯丹、柠檬酸哌嗪、哌嗪二盐酸盐、盐酸哌嗪、哌嗪单盐酸盐、磷酸哌嗪、哌嗪二硫化碳络合物、盐酸吡利霉素、垂体促黄体生成激素、泊洛扎林、硫酸多粘菌素b、聚氧乙烯23十二烷基醚、多硫酸化糖胺多糖、泊那珠利、猪胰岛素锌、猪垂体分泌的性促卵泡激素、泊沙康唑、钾、柠檬酸钾、磷酸钾、普多沙星、氯磷定、吡喹酮、泼尼松龙、醋酸泼尼松龙、磷酸泼尼松龙钠、琥珀酸泼尼松龙钠、泼尼松龙叔醋酸丁酯、泼尼松、扑米酮、二马来酸丙氯拉嗪、乙二磺酸丙氯拉嗪、马来酸丙氯拉嗪、黄体酮、盐酸氯丙嗪、盐酸丙美卡因、丙酰丙嗪盐酸盐(propiopromazine hydrochloride)、丙泊酚、前列他林、双羟萘酸噻嘧啶、酒石酸噻嘧啶、马来酸新安替根、乙嘧啶、rabacfosadine、盐酸莱克多巴胺、罗贝考昔、盐酸氯苯胍、盐酸罗米非定、洛克沙胂、盐霉素钠、sarolaner、司可巴比妥钠、西拉菌素(selamectin)、盐酸司来吉兰、二硫化硒、西姆卓霉素钠、西姆卓霉素钠生物质、血清促性腺激素、七氟醚、磺胺嘧啶银、氯化钠、亚硒酸钠、磺胺氯吡嗪钠一水合物、磺胺氯哒嗪钠、磺胺二甲嘧啶钠、大观霉素、大观霉素二盐酸盐五水合物、大观霉素盐酸盐五水合物、大观霉素硫酸盐四水合物、多杀菌素、康力龙、硫酸链霉素、磺胺溴甲嘧啶钠、磺胺氯哒嗪、磺胺嘧啶、磺胺嘧啶钠、磺胺地索辛、磺胺乙氧嗪(sulfaethoxypyridazine)、磺胺甲嘧啶、磺胺二甲嘧啶、硫酸氢磺胺二甲嘧啶、磺胺甲二唑、磺胺硝苯、磺胺喹沙啉、磺胺喹沙啉钠、磺胺二甲异噁唑、磺粘菌素、替泊沙林、特比萘芬、丙酸睾酮、盐酸丁卡因、四环素、盐酸四环素、磷酸四环素、噻苯铵醚(thenium closylate)、噻苯咪唑、硫烯丙巴比妥钠(thialbarbitonesodium)、硫戊巴比妥钠、硫喷妥钠、硫链丝菌肽、促甲状腺激素、泰妙菌素、富马酸氢泰妙菌素、替卡西林二钠、泰地罗新(tildipirosin)、盐酸替来他明、磷酸替米考星、替鲁膦酸二钠、噻昔达唑(tioxidazole)、磷酸托西尼布(toceranib phosphate)、盐酸妥拉唑啉、托萘酯、甲苯、群勃龙醋酸酯、曲安奈德、甲磺酸三卡因、敌百虫、三氯甲噻嗪、盐酸三氟丙嗪、曲洛司坦、酒石酸阿利马嗪(trimeprazine tartrate)、甲氧苄啶、盐酸曲吡那敏、醋酸曲普瑞林、胰蛋白酶、泰拉霉素、泰乐菌素、磷酸泰乐菌素、酒石酸泰乐菌素、泰乐菌素(tylvalosin)、酒石酸泰万菌素(tylvalosin tartrate)、克霉唑、维吉尼亚霉素、维生素E、甲苯噻嗪、盐酸甲苯噻嗪、盐酸育亨宾、折仑诺、齐帕特罗、盐酸齐帕特罗、葡萄糖酸锌、球痢灵和盐酸唑拉西泮。
在一些实施例中,活性剂可以是用于针对以下进行免疫的抗原:疟疾、狂犬病、炭疽、肺结核、乙型肝炎、白喉、百日咳、破伤风、乙型流感嗜血杆菌、脊髓灰质炎病毒、麻疹、腮腺炎、风疹、水痘、肺炎球菌、甲型肝炎、流感及其组合。
在一些实施例中,活性剂可以是蛋白质、肽、抗原、抗体、抗体片段、核酸、染料、放射性示踪剂、造影剂、有机化合物或无机化合物。
抗原/抗体片段
在一些实施例中,活性剂可以是抗原。抗原可以源自可以感染受试者的病原体。因此,抗原可以源自例如细菌、病毒、真菌或寄生虫。抗原可以是肿瘤抗原。抗原可以是过敏原,包括但不限于花粉、动物皮屑、霉菌、尘螨、跳蚤过敏原、唾液过敏原、草或食物(例如花生和其他坚果)。抗原可以是自身抗原。自身抗原可以与自身免疫性疾病相关,例如像胰岛抗原。
细菌抗原的非限制性实例包括源自以下的抗原:炭疽、弯曲杆菌属、霍乱弧菌(Vibrio cholera)、梭菌纲(包括艰难梭菌(Clostridium difficile))、白喉、肠出血性大肠杆菌、产肠毒素性大肠杆菌、贾弟虫属、淋球菌、幽门螺杆菌(Helicobacter pylori)、乙型流感嗜血杆菌、非分型流感嗜血杆菌、军团菌属、脑膜炎球菌、分枝杆菌(包括那些负责结核病、百日咳的生物体)、肺炎球菌属、沙门氏菌属、志贺氏菌属、葡萄球菌属、A组β-溶血性链球菌、B链球菌、破伤风、伯氏疏螺旋体(Borrelia burgdorfi)、耶尔森菌属等。根据本发明,细菌抗原包括例如毒素、类毒素(即,化学灭活的毒素,毒性较小但保留免疫原性)、其亚基或其组合、以及毒力或定植因子。细菌成分、产物、裂解物和/或提取物可用作细菌抗原的来源。
抗原可以源自病毒。病毒抗原包括但不限于源自以下的抗原:腺病毒、登革热血清型1至4病毒、埃博拉病毒、肠病毒、汉坦病毒、肝炎病毒血清型A至E、单纯疱疹病毒1或2、人免疫缺陷病毒、人乳头瘤病毒、流感病毒、麻疹(measles/rubeola)病毒、日本马脑炎病毒、乳头瘤病毒、细小病毒B19、脊髓灰质炎病毒、狂犬病病毒、呼吸道合胞病毒、轮状病毒、圣路易斯脑炎病毒、牛痘病毒、黄热病病毒、风疹病毒、水痘(chickenpox)病毒、水痘(varicella)病毒和腮腺炎病毒。病毒成分、产物、裂解物和/或提取物可以用作病毒抗原的来源。
在一些实施例中,抗原可以是肿瘤抗原。肿瘤抗原包括但不限于胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤。
在一些实施例中,活性剂可以是佐剂。术语“佐剂”是指用于特异性或非特异性增强抗原特异性免疫应答的物质。术语“佐剂活性”是通过将佐剂包含在组合物中或作为方法的一部分来增强对抗原(即,作为与佐剂分离的化学结构的抗原)的免疫应答的能力。佐剂包括但不限于油乳剂(例如完全或不完全的弗洛伊德(Freud)佐剂);趋化因子(例如,防御素、HCC-1、HCC-4、MCP-1、MCP-3、MCP-4、MIP-1α、MIP-1β、MIP-1δ、MIP-3α和MIP-2);趋化因子受体的其他配体(例如,CCR-1、CCR-2、CCR-5、CCR-6、CXCR-1);细胞因子(例如,IL-1、IL-2、IL-6、IL-8、IL-10、IL-12、IFN-γ、TNF-α、GM-CSF);这些细胞因子、细菌DNA或寡核苷酸的免疫刺激性CpG基序的受体的其他配体;胞壁酰二肽(MDP)及其衍生物(例如,莫拉丁酯,苏氨酰-MDP,胞壁酰三肽);热休克蛋白及其衍生物;利什曼原虫同源物及其衍生物;细菌ADP-核糖基化外毒素、其化学缀合物及其衍生物(例如,遗传突变体、含有A和/或B亚基的片段、化学类毒素形式);或盐(例如氢氧化铝或磷酸铝、磷酸钙)。
在一些实施例中,活性剂可以是一种或多种抗体和抗体片段。例如,该抗体可以是治疗性抗体或诊断性抗体。治疗性抗体包括但不限于识别胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤的抗原的抗体。实例包括但不限于阿昔单抗;阿巴西普(abatacept);阿达木单抗(adalimumab);阿利鲁单抗(abrilumab);阿托珠单抗(afutuzumab);阿柏西普(aflibercept);阿仑单抗(alemtuzumab);阿法赛特(alefacept);阿拉赛珠单抗-培戈(alacizumab pegol);阿那白滞素;阿西莫单抗(arcitumomab);阿塞西普(atacicept);阿替珠单抗(atlizumab);阿托木单抗(atorolimumab);巴利昔单抗(basiliximab);巴奈西普(baminercept);贝妥莫单抗(bectumomab);贝利木单抗(belimumab);贝索单抗(besilesomab);贝伐单抗(bevacizumab);比西单抗(biciromab);贝拉西普(belatacept);贝伦妥单抗-维多汀(brentuximab vedotin);布洛鲁单抗(brodalumab);康纳单抗(canakinumab);卡罗单抗喷地肽(capromab pendetide);卡妥索单抗(catumaxomab);赛妥珠单抗-培戈(certolizumabpegol);西妥昔单抗(cetuximab);克利妥珠单抗-泰坦(clivatuzumab tetraxetan);达利珠单抗(daclizumab);地诺单抗(denosumab);依库丽单抗(eculizumab);依决洛单抗(edrecolomab);依法珠单抗(efalizumab);依芬谷单抗(efungumab);Eloctate;厄马索单抗(ertumaxomab);依那西普(etanercept);依他赛珠单抗(etaracizumab);法索单抗(fanolesomab);法乐妥珠单抗(farletuzumab);芳妥珠单抗(fontolizumab);吉妥珠单抗奥佐米星(gemtuzumab ozogamicin);吉仑妥昔单抗(girentuximab);戈利木单抗(golimumab);替伊莫单抗-替坦(ibritumomab tiuxetan);伊戈伏单抗(igovomab);英西单抗(imciromab);英利昔单抗(infliximab);伊匹单抗(ipilimumab);拉贝珠单抗(labetuzumab);美泊利单抗(mepolizumab);莫他珠单抗(motavizumab);莫罗单抗(muromonab)-CD3;那他珠单抗(natalizumab);尼妥珠单抗(nimotuzumab);巯诺莫单抗-莫坦(Nofetumomab merpentan);奥比妥珠单抗(obinutuzumab);奥法木单抗(ofatumumab);奥马珠单抗(omalizumab);奥戈伏单抗(oregovomab);帕利珠单抗(palivizumab);帕尼单抗(panitumumab);培妥莫单抗(pemtumomab);帕妥珠单抗(pertuzumab);雷莫芦单抗(ramucirumab);兰尼单抗(ranibizumab);雷昔巴库单抗(raxibacumab);利妥昔单抗(rituximab);利纳西普(rilonacept);罗维珠单抗(rovelizumab);卢普利珠单抗(ruplizumab);硫索单抗(sulesomab);他卡妥珠单抗-泰坦(Tacatuzumab tetraxetan);特非巴珠单抗(tefibazumab);托珠单抗(tocilizumab);曲妥珠单抗(trastuzumab);阿多-曲妥珠单抗恩他新(Ado-Trastuzumab Emtansine);托西莫单抗(tositumomab);TRBS07;优特吉努单抗(ustekinumab);维多利珠单抗(vedolizumab);威司利珠单抗(visilizumab);伏妥莫单抗(votumumab);扎鲁妥木单抗(zalutumumab);扎诺利木单抗(Zanolimumab)及其片段。
在一些实施例中,活性剂可以是诊断性抗体。诊断性抗体包括但不限于针对肿瘤抗原、癌症抗原、过敏原、细菌抗原、病毒抗原、药物、激素、植物凝集素、内毒素及其组合的抗体。在一些实施例中,使用本领域已知的方法(包括但不限于碳二亚胺缀合、酯化、高碘酸钠氧化然后还原性烷基化以及戊二醛交联),还可以将所公开的抗体及其片段与毒素、化学治疗药物、放射性标记偶联。
诊断性抗体及其片段可以包括用于检测的可检测部分。术语“可检测部分”是指在抗体中掺入另一种分子,例如但不限于发色团、荧光部分、酶、抗原、具有特定反应性的基团、化学发光部分和电化学可检测部分等。在一些实施例中,抗体是生物素酰化的。
在一些实施例中,本文公开的抗体及其片段可以用可检测部分进行标记。可检测部分可以是能够直接或间接产生可检测信号的任何部分。针对抗体的可检测部分的实例包括但不限于以下:放射性同位素或放射性核素(例如3H、14C、15N、35S、90Y、99Tc、111In、125I、131I)、荧光标记(例如FITC、罗丹明、镧系元素磷)、酶标记(例如辣根过氧化物酶、β-半乳糖苷酶、萤光素酶、碱性磷酸酶)、化学发光标记、生物素基基团、由二级报告子识别的预定多肽表位(例如亮氨酸拉链成对序列、二级抗体的结合位点、金属结合结构域、表位标签)、磁性剂(例如钆螯合物)、毒素(例如百日咳毒素)、紫杉醇、细胞松弛素B、短杆菌肽D、溴化乙锭、吐根碱、丝裂霉素、依托泊苷、替尼泊苷、长春新碱、长春碱、秋水仙素、阿霉素、道诺霉素、二羟基炭疽菌素二酮、米托蒽醌、光神霉素、放线菌素D、1-去氢睾酮、糖皮质激素、普鲁卡因、丁卡因、利多卡因、普萘洛尔和嘌呤霉素及其类似物或同系物。在一些实施例中,标记通过各种长度的间隔臂附接以减少潜在的空间位阻。这样的抗体及其片段可以用于诊断性应用,包括但不限于检测应用和成像应用。
在一些实施例中,可检测部分包含荧光团。任何荧光团可以与本文公开的任何抗体一起使用,条件是荧光团的缀合产生在体内(例如,在施用于受试者后)和/或体外可检测的组合物,并且不会对抗体片段结合其表位的能力产生负面影响。代表性的荧光团包括但不限于7-二甲基氨基香豆素-3-甲酸、丹磺酰氯、硝基苯并二唑胺(NBD)、达布酰氯、肉桂酸、荧光素羧酸、尼罗蓝、四甲基羧基罗丹明、四乙基磺基罗丹明、5-羧基-X-罗丹明(5-ROX)和6-羧基-X-罗丹明(6-ROX)。应当理解,这些代表性的荧光团仅是示例性的,并且还可以使其他的荧光团。例如,ALEXA染料系列包括至少19种不同的染料,这些染料的特征在于不同的发射光谱。这些染料包括ALEXA350、405、430、488、500、514、532、546、555、568、594、610、633、635、647、660、680、700和750(可从美国加利福尼亚州卡尔斯巴德市英杰公司(Invitrogen Corp.,Carlsbad,California,United States of America)获得),并且本领域技术人员可在考虑本说明书后基于以下标准选择使用哪种染料,这些标准包括但不限于特定ALEXA的化学组成、是否要使用多个可检测部分以及每个可检测部分的发射光谱、要使用的检测技术等。
在一些实施例中,可检测部分包含花菁染料。可以与本公开主题的抗体片段缀合的花菁染料的非限制性实例包括由安玛西亚生物科学公司(Amersham Biosciences)(美国新泽西州皮斯卡塔韦(Piscataway,New Jersey,United States of America))提供的琥珀酰亚胺酯CyS、CyS.5和Cy7。
在一些实施例中,可检测部分包含近红外(NIR)染料。可以与本公开主题的抗体片段缀合的近红外染料的非限制性实例包括NIR641、NIR664、NIT7000和NIT782。
在一些实施例中,使用第二抗体检测生物素酰化的抗体,该第二抗体包含抗生物素蛋白或链霉抗生物素蛋白基团并且还与荧光标记缀合,该荧光标记包括但不限于Cy3、Cy5、Cy7、以及可获得自INVITROGENTM(美国加利福尼亚州卡尔斯巴德市)的ALEXA系列荧光标记中的任何一个。在一些实施例中,抗体片段直接用荧光标记进行标记,并且通过荧光激活细胞分选术分离与该抗体片段结合的细胞。另外的检测策略是本领域技术人员已知的。
示踪剂/造影剂
在一些实施例中,活性剂可以是放射性示踪剂,例如但不限于241Am、51Cr、60Co、64Cu、18F、67Ga、198Au、113In、111In、131I、125I、59Fe、85Kr、197Hg、32P、42K、75Se、24Na、99Tc、133Xe、90Y、3H、14C、35S、15N和169Y。
在一些实施例中,活性剂可以是用于成像的造影剂。造影剂的非限制性实例包括不透射线的造影剂、顺磁性造影剂、超顺磁性造影剂、光学成像部分、CT造影剂和其他造影剂。例如,不透射线的造影剂(用于X射线成像)将包括无机和有机碘化合物(例如泛影酸盐)、不透射线的金属及其盐(例如银、金、铂等)以及其他不透射线的化合物(例如钙盐、钡盐如硫酸钡、钽和氧化钽)。合适的顺磁性造影剂(用于MR成像)包括二亚乙基三胺五乙酸钆(Gd-DTPA)及其衍生物,以及其他钆、锰、铁,镝、铜、铕、铒、铬、镍和钴的复合物,包括与1,4,7,10-四氮杂环十二烷-N,N′,N″,N′″-四乙酸(DOTA)、乙二胺四乙酸(EDTA)、1,4,7,10-四氮杂环癸烷-N,N′,N″-三乙酸(DO3A)、1,4,7-三氮杂环壬烷-N,N′,N″-三乙酸(NOTA)、1,4,8,11-四氮杂环十四烷-N,N′,N″,N′″-四乙酸(TETA)、羟基苄基乙二胺二乙酸(HBED)等的复合物。合适的超顺磁性造影剂(用于MR成像)包括磁铁矿、超顺磁性氧化铁、单晶氧化铁,特别是可以与带负电荷主链缀合的这些试剂的每一种的复合形式。另外,其他合适的显像剂是CT造影剂(包括碘化和非碘化的离子和非离子CT造影剂)、以及例如自旋标记等造影剂或其他诊断有效的试剂。组合物中可能存在的其他染色剂和染料包括但不限于刚果红、酸性品红、吖啶橙、碱性品红、吉姆沙染色剂、甲基绿、亚甲蓝、中性红、橙-G和地衣红。
神经毒素
在一些实施例中,活性剂是神经毒素。在一些实施例中,神经毒素剂可以选自由以下组成的组:A型肉毒杆菌毒素、B型肉毒杆菌毒素、C型肉毒杆菌毒素、D型肉毒杆菌毒素、E型肉毒杆菌毒素、F型肉毒杆菌毒素、abobotulinumtoxinA、BTX-A、daxibotulinumtoxinA、incobotulinumtoxinA、onabotulinumtoxinA、rimabotulinumtoxinB、突触融合蛋白、其衍生物(包括但不限于工程化重组神经毒素,包括如WO 2014/068317和WO 199807864(将其通过引用以其全文特此并入)所述的工程化重组肉毒杆菌毒素或其片段)、其药学上可接受的盐及其组合。
这些组合物可以包括一种或多种占总组合物的约0.1wt.%至约25wt.%、约0.1wt.%至约20wt.%、约0.1wt.%至约15wt.%、约0.1wt.%至约10wt.%、约0.1wt.%至约8wt.%、约0.1wt.%至约5wt.%、约0.1wt.%至约4wt.%、约0.1wt.%至约3wt.%、约0.1wt.%至约3wt.%、或约0.1wt.%至约1wt.%的活性剂。具体实例包括约0.1wt.%、约0.5wt.%、约1wt.%、约2wt.%、约5wt.%、约10wt.%、约25wt.%、以及介于这些值中的任何两个值之间的范围。相对于组合物的总量,本文公开的重量百分比可以是重量/重量百分比或重量/体积百分比。
在一些实施例中,活性剂能以约1微克至约100毫克/mL组合物、约1微克至约10毫克/mL组合物、约1微克至约5毫克/mL组合物、约1微克至约1毫克/mL组合物、或约1微克至约100微克/mL组合物存在。
在一些实施例中,组合物中活性剂如神经毒素剂的有效量为0.1个单位、0.2个单位、0.3个单位、0.4个单位、0.5个单位、0.6个单位、0.7个单位、0.8个单位、0.8个单位、0.9个单位、1.0unit,2个单位、3,个单位、4个单位、5个单位、6个单位、7个单位、8个单位、9个单位、10个单位、11个单位、12个单位、13个单位、14个单位、15个单位、16个单位、17个单位、18个单位、19个单位、20个单位、25个单位、30个单位、35个单位、40个单位、45个单位、50个单位、100个单位、125个单位、150个单位、175个单位、200个单位、225个单位、250个单位、275个单位、300个单位、325个单位、350个单位、375个单位、400个单位、425个单位、450个单位、475个单位、500个单位、以及介于这些值中的任何两个值之间的任何单个量或任何范围。
本发明涵盖的组合物的特定实例包括以下组合物,这些组合物含有约0.01wt.%至约2.0wt.%的一种或多种诱饵分子(平均分子量为约2,000Da至约20,000Da)以及活性剂(如米诺环素、水杨酸盐、利多卡因、防晒剂、视黄醇、比马前列素、类固醇、克林霉素、米诺地尔、烟酰胺、以及相似大小的活性剂及其组合)。本发明涵盖的组合物的其他实例包括以下组合物,这些组合物含有约0.5wt.%至约5.0wt.%的一种或多种诱饵分子(平均分子量为约2,000Da至约30,000Da)以及一种或多种活性剂(如抗生素、抗真菌剂、生物制剂、抗体、大分子活性剂、基于肽的治疗剂、以及相似大小的活性剂及其组合)。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的细胞外基质组分、其片段及其组合(诱饵分子)。
在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的一种或多种诱饵分子(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的一种或多种诱饵分子(平均分子量为约2,000Da至约30,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的细胞外基质组分、其片段及其组合(诱饵分子)。
在一些实施例中,组合物包括约0.01wt.%至约2.0wt.%的弹性蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的弹性蛋白肽片段(平均分子量为约2,000Da至约30,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,组合物包括约0.1wt.%至约1.5wt.%的弹性蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约1wt.%的弹性蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约0.5wt.%的弹性蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的弹性蛋白肽片段。
在一些实施例中,组合物包括约0.01wt.%至约2.0wt.%的透明质酸(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的透明质酸(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,组合物包括约0.1wt.%至约1.5wt.%的透明质酸(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约1.0wt.%的透明质酸(平均分子量为约2,000Da至约60,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约0.5wt.%的透明质酸(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的透明质酸片段。
在一些实施例中,组合物包括约0.01wt.%至约2.0wt.%的胶原蛋白片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的胶原蛋白片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,组合物包括约0.1wt.%至约1.5wt.%的胶原蛋白片段(平均分子量为约2,000Da至约30,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约1.0wt.%的胶原蛋白片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约0.5wt.%的胶原蛋白片段(平均分子量为约2,000Da至约30,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的胶原蛋白片段。
在一些实施例中,组合物包括约0.01wt.%至约2.0wt.%的纤连蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的纤连蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,组合物包括约0.1wt.%至约1.5wt.%的纤连蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约1.0wt.%的纤连蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约0.5wt.%的纤连蛋白肽片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的纤连蛋白肽片段。
在一些实施例中,组合物包括约0.01wt.%至约2.0wt.%的凝集素片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的一种或多种本文公开的任何活性剂。在一些实施例中,组合物包括约0.1wt.%至约2.0wt.%的凝集素片段(平均分子量为约2,000Da至约30,000Da)以及约0.1wt.%至约5wt.%的活性剂,该活性剂选自由以下组成的组:水杨酸盐、米诺环素、利多卡因、克林霉素、米诺地尔、烟酰胺、地仑丁、丙酸视黄酯、环苯扎林、西地那非、阿那曲唑、抗坏血酸、氨甲环酸、对苯二酚、克霉唑、renokin、托萘酯、特比萘芬、强的松、维甲酸、防晒剂、视黄醇、比马前列素、类固醇、加巴喷丁、沙瑞环素、苯妥英、乙酰水杨酸、硫酸庆大霉素、丙酸氯倍他索、曲酸、咪喹莫特、抗体如艾塞吉珠单抗、维生素、萘普生、抗氧化剂、及其组合。在一些实施例中,组合物包括约0.1wt.%至约1.5wt.%的凝集素片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约1.0wt.%的凝集素片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,组合物包括约0.1wt.%至约0.5wt.%的凝集素片段(平均分子量为约2,000Da至约20,000Da)以及约0.1wt.%至约5wt.%的活性剂。在一些实施例中,这些组合物可以基本上不包括分子量大于60,000Da的凝集素片段。
感觉改变剂
在一些实施例中,活性剂是选自由以下组成的组的感觉改变剂:冷却剂、温热剂、松弛剂或舒缓剂、刺激剂或提神剂(refreshing agent)、及其混合物。这些感觉改变剂可以与诱饵分子一起局部施用或递送。这些组合物可以配制成糊剂、凝胶、软膏、洗剂、乳剂、霜剂、泡沫、摩丝、液体、喷雾剂、悬浮剂、分散剂、粉末或气雾剂。具有感觉改变剂的组合物可以施用于如本文所公开的任何表面组织。
在一些实施例中,冷却剂选自但不限于:薄荷醇;薄荷醇的异构体、薄荷醇衍生物;4-甲基-3-(1-吡咯烷基)-2[5H]-呋喃酮;WS-23、依色林(Icilin)、依色林联合利华类似物、5-甲基-4-(1-吡咯烷基)-3-[2H]-呋喃酮;4,5-二甲基-3-(1-吡咯烷基)-2[5H]-呋喃酮;异薄勒醇、3-(l-薄荷氧基)丙烷-1,2-二醇、3-(l-薄荷氧基)-2-甲基丙烷-1,2-二醇、对薄荷烷-2,3-二醇、对薄荷烷-3,8-二醇、6-异丙基-9-甲基-1,4-二氧杂-螺[4,5]癸烷-2-甲醇、琥珀酸薄荷酯及其碱土金属盐、三甲基环己醇、N-乙基-2-异丙基-5-甲基环己烷甲酰胺、日本薄荷(薄荷(Mentha arvensis))油、薄荷油(peppermint oil)、薄荷酮、薄荷酮甘油缩酮、乳酸薄荷酯、3-(l-薄荷氧基)乙-1-醇,3-(l-薄荷氧基)丙-1-醇、3-(l-薄荷氧基)丁-1-醇、l-薄荷基乙酸N-乙酰胺、l-薄荷基-4-羟基戊酸酯、l-薄荷基-3-羟基丁酸酯、N,2,3-三甲基-2-(1-甲基乙基)-丁酰胺和留兰香油。
在一些实施例中,温热剂选自但不限于:多元醇、辣椒素、辣椒粉、辣椒酊、辣椒提取物、辣椒素、北美金缕梅(hamamalis)、高辣椒碱、高二氢辣椒碱、壬酰基香草基酰胺、壬酸香草基醚、香草醇烷基醚衍生物(例如香草基乙醚、香草基丁醚、香草基戊醚和香草基己醚)、异香草醇烷基醚、乙基香草醇烷基醚、藜芦醇衍生物、取代的苯甲醇衍生物、取代的苯甲醇烷基醚、香草醛丙二醇缩醛、乙基香兰醛丙二醇缩醛、姜提取物、姜油、姜酚和姜酮。
在一些实施例中,松弛剂或舒缓剂选自但不限于:草本植物提取物,其选自由芦荟、α没药醇、D-泛醇、尿囊素、金缕梅属、甘菊、蓍草组成的组;金盏花属提取物、紫草科植物提取物、金缕梅(witch hazel)和其他收敛剂提取物、海草和燕麦提取物;油,其选自由杏仁油、鳄梨油和紫草科植物组成的组;以及精油,其选自由豆蔻、桉树、欧薄荷(menthapiperita)(薄荷(peppermint))、海索草和迷迭香组成的组;蜡质或油性物质,其选自由羊毛脂或凡士林胶状物组成的组;矿物质,其选自由氧化锌、炉甘石和硒组成的组;维生素,其选自由生育酚乙酸酯(维生素E)组成的组;以及药剂,其选自由镇痛剂、麻醉剂、抗炎剂和抗组胺药、以及肌肉松弛剂组成的组;薄荷醇、樟脑、丁香酚、桉油精、黄樟素、水杨酸甲酯、乳酸薄荷酯、乙氧基乙酸薄荷酯、薄荷酮甘油缩醛、3-1-薄荷氧基丙烷-1,2-二醇、1-薄荷基碳酸乙酯、(1S,3S,4R)-对-薄荷-8-烯-3-醇、薄荷基吡咯烷酮甲酸酯、N-取代-对-薄荷烷-3-甲酰胺金缕梅属提取物和姜油。
在一些实施例中,刺激剂或提神剂选自但不限于:醇、L-薄荷醇、樟脑、薄荷油、辣椒提取物、辣椒素、烟酸苄酯、水杨酸盐、乙二醇水杨酸酯、乙酰胆碱、血清素、组胺、前列腺素、神经递质;CNS刺激剂、咖啡因和奎宁。
治疗性细胞
在一些实施例中,活性剂是治疗性细胞。在一些实施例中,治疗性细胞是干细胞、基因工程化的哺乳动物细胞、抗原呈递细胞或其组合。
在一些实施例中,治疗性细胞可以是干细胞。例如,干细胞可以是成年干细胞、组织特异性干细胞、胎儿干细胞、脐带血干细胞、源自脐带的细胞、胚胎干细胞、诱导性多能干细胞、多能(pluripotent)间充质干细胞、多潜能(multipotent)间充质干细胞、造血干细胞及其组合。在一些实施例中,间充质干细胞(MSC)、多潜能成年祖细胞(MAPC)和/或其他干细胞能够分化成组织或器官系统的专门或部分专门的细胞类型。在一些实施例中,对MSC、MAPC和/或其他干细胞进行基因工程化或修饰以过表达趋化因子和/或趋化因子受体,这可以显著改善经基因修饰的MSC、MAPC、和/或其他干细胞的存活能力和寿命,并且潜在地改善其中引入了经基因修饰的干细胞的组织的存活能力。当将基因修饰的干细胞引入哺乳动物受试者用于治疗性应用和/或细胞疗法时,过表达的趋化因子和/或趋化因子受体可以减轻经基因修饰的干细胞的凋亡。
在一些实施例中,治疗性细胞是基因工程化的哺乳动物细胞。可将基因工程化的哺乳动物细胞配置为通过稳定整合进染色体中的质粒构建体、病毒构建体或通过本领域已知的其他技术表达任何内源或外源基因。例如,可以对哺乳动物细胞进行基因工程化以表达肿瘤或癌症抗原。在一些实施例中,肿瘤抗原包括但不限于胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤。在一些实施例中,基因工程化的哺乳动物细胞可以是皮肤细胞。
在一些实施例中,治疗性细胞可以是抗原呈递细胞,例如T细胞、B细胞、树突状细胞和巨噬细胞。在一些实施例中,治疗性细胞是表达嵌合受体(例如嵌合抗原受体(CAR)和转基因T细胞受体(TCR))的工程化免疫细胞。嵌合受体(例如CAR或转基因TCR)通常结合一种或多种抗原,该抗原由受试者的疾病或病症和/或其一种或多种细胞或组织表达,与受试者的疾病或病症和/或其一种或多种细胞或组织相关,和/或对受试者的疾病或病症和/或其一种或多种细胞或组织具有特异性。此类疾病可包括肿瘤、癌症、其他增殖性疾病、自身免疫性疾病或障碍、和/或传染剂或传染病。
在一些实施例中,治疗性细胞可以是例如成纤维细胞、体被细胞、脂肪细胞、前脂肪细胞、干细胞、上皮细胞、视网膜细胞、免疫功能细胞、黑素细胞或其他色素细胞、毛囊细胞、角质形成细胞、朗格汉斯细胞、默克尔细胞、肥大细胞、肌肉细胞或其组合。还考虑了递送肌肉细胞、骨细胞、胰腺细胞、粘膜细胞、软骨细胞、神经系统细胞、激素分泌细胞、内分泌细胞、肠细胞和生殖细胞的方法。
在一些实施例中,治疗性细胞是自体细胞。能以多种方式从组织中分离自体细胞,所有这些方式都是本领域技术人员已知的。在一个实施例中,可以通过常规方法从活检材料中分离细胞。可以从患者的与缺陷的组织类型或组织再生有关的任何组织中提取活检材料。举例来说而非限制,合适的细胞包括肌腱细胞、肌细胞、干细胞、骨细胞、软骨细胞、上皮细胞、角质形成细胞、神经细胞(包括但不限于神经细胞、星形胶质细胞、树突状细胞和神经胶质细胞)、成纤维细胞、齿状细胞(odontocyte)、滑膜细胞、脂肪细胞和牙骨质细胞。另外,这些细胞类型的前体细胞也可用于本发明的组合物中。在一个实施例中,例如,成肌细胞(肌细胞的前体)、成骨细胞(骨细胞的前体)、以及成神经细胞(神经细胞的前体)都可用于本公开中。在一些实施例中,细胞和细胞前体是自体细胞和自体细胞前体。
在一些实施例中,治疗性细胞是同种异体细胞。术语“同种异体的”是指具有不同基因组成的细胞、组织或生物体。本文公开的干细胞和基因工程化的哺乳动物细胞可以是同种异体细胞。
在一些实施例中,组合物中治疗性细胞的数量可以是约102个细胞至约106个细胞/mL组合物、约103个细胞至约106个细胞/mL组合物、约104个细胞至约106个细胞/mL组合物、约105个细胞至约106个细胞/mL组合物、约103个细胞至约109个细胞/mL组合物、或约103个细胞至约1010个细胞/mL组合物。
组合物
各种实施例的组合物可以包括有效量的以上鉴定的任何活性剂或其组合、以及一种或多种本文公开的任何诱饵分子。本文公开的组合物被配制成用于局部施用、透皮(transdermal)施用、经皮(percutaneous)施用或通过微针注射至受试者的皮肤。无意限制,但出于示例性实施例的目的,考虑到组合物可以是糊剂、凝胶、软膏、洗剂、乳剂、霜剂、泡沫、摩丝、液体、喷雾剂、悬浮剂、分散剂、粉末或气雾剂。该组合物包括一种或多种赋形剂,以提供所需的形式和所需的粘度、流动性或其他物理或化学特性,以有效地施加、覆盖和粘附于皮肤/表面组织。
在一些实施例中,本文所述的组合物可以进一步包括一种或多种化妆品或药学上可接受的稀释剂、填充剂、崩解剂、粘合剂、润滑剂、表面活性剂、疏水性媒介物、水溶性媒介物、乳化剂、缓冲剂、湿润剂、保湿剂、增溶剂、防腐剂、着色剂、增塑剂、载体、赋形剂等及其组合。本领域普通技术人员可以参考各种药理学参考文献,例如像,Modern Pharmaceutics[现代药剂学],Banker和Rhodes,Marcel Dekker,Inc.[马塞尔·德克尔公司](1979)以及Goodman和Gilman的The Pharmaceutical Basis of Therapeutics[治疗学的药理学基础],第6版,MacMillan Publishing Co[麦克米伦出版公司],纽约(1980),它们用于指导确定此类组分在实施例的组合物中的量。
基于预期的配制品类型选择组合物中的赋形剂。标准赋形剂包括明胶、酪蛋白、卵磷脂、阿拉伯树胶、胆固醇、黄芪胶、硬脂酸、苯扎氯铵、硬脂酸钙、单硬脂酸甘油酯、鲸蜡硬脂醇、聚西托醇乳化蜡、山梨聚糖酯、聚氧乙烯烷基醚、聚氧乙烯蓖麻油衍生物、聚氧乙烯山梨聚糖脂肪酸酯、聚乙二醇、聚氧乙烯硬脂酸酯、胶体二氧化硅、磷酸盐、十二烷基硫酸钠、羧甲基纤维素钙、羧甲基纤维素钠、甲基纤维素、羟乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素邻苯二甲酸酯、非结晶纤维素、硅酸镁铝、三乙醇胺、聚乙烯醇、聚乙烯吡咯烷酮、糖和淀粉。
“表面活性剂”是降低表面张力并从而增加产品的乳化、起泡、分散、铺展和润湿性能的表面活性剂。合适的非离子表面活性剂包括乳化蜡、单油酸甘油酯、聚氧乙烯烷基醚、聚氧乙烯蓖麻油衍生物、聚山梨酸酯、山梨聚糖酯、苯甲醇、苯甲酸苄酯、环糊精、单硬脂酸甘油酯、泊洛沙姆、聚维酮或其组合。在一个实施例中,非离子表面活性剂是硬脂醇。
“乳化剂”是表面活性物质,其促进一种液体在另一种液体中悬浮并促进油和水的稳定混合物或乳液的形成。常见的乳化剂是金属皂、某些动植物油和各种极性化合物。合适的乳化剂包括阿拉伯胶、阴离子乳化蜡、硬脂酸钙、卡波姆、鲸蜡硬脂醇、鲸蜡醇、胆固醇、二乙醇胺、棕榈硬脂酸乙二醇酯、单硬脂酸甘油酯、单油酸甘油酯、羟丙基纤维素、羟丙甲纤维素、羊毛脂、含水物、羊毛脂醇、卵磷脂、中链甘油三酯、甲基纤维素、矿物油和羊毛脂醇、磷酸二氢钠、单乙醇胺、非离子乳化蜡、油酸、泊洛沙姆、泊洛沙姆、聚氧乙烯烷基醚、聚氧乙烯蓖麻油衍生物、聚氧乙烯山梨聚糖脂肪酸酯、聚氧乙烯硬脂酸酯、藻酸丙二醇酯、自乳化单硬脂酸甘油酯、脱水柠檬酸钠、十二烷基硫酸钠、山梨聚糖酯、硬脂酸、葵花油、黄芪胶、三乙醇胺、黄原胶或其组合。在一个实施例中,乳化剂是硬脂酸甘油酯。
在一些实施例中,本文所述的组合物可以被配制成液体。用于局部施用的液体剂型可以包括稀释剂,例如像,醇、二醇、油、水等。这样的组合物还可包括润湿剂或乳化剂。在一些实施例中,实施例的组合物可以被配制成水包油或油包水乳剂。霜剂可以是其中水相分散在油相中的油包水(w/o)乳剂,或是其中油分散在水相中的水包油(o/w)乳剂。软膏通常是指更具粘性的水包油霜剂。传统的软膏基质(即,载体)包括烃类(矿脂、蜂蜡等)、植物油、脂肪醇(胆固醇、羊毛脂、羊毛醇、硬脂醇等)或硅酮。不溶性固体(例如淀粉、氧化锌、碳酸钙或滑石)也可以用于软膏和霜剂中。上述组合物的凝胶形式可以通过将大量的水性液体或水性醇液体截留在聚合物或胶态固体颗粒的网络中而形成。这样的聚合物或胶体(胶凝剂或增稠剂)的浓度通常低于10%w/w,并且其包括羧甲基纤维素、羟丙基甲基纤维素、羟乙基纤维素、甲基纤维素、藻酸钠、藻酸、果胶、黄芪胶、角叉菜、琼脂、粘土、硅酸铝、卡波姆等。
乳剂是一种液体以小液滴散布在第二种液体的整个主体内的一种制剂。经分散的液体是不连续相,而分散介质是连续相。当油是经分散的液体且水溶液是连续相时,称为水包油乳剂,而当水或水溶液是分散相且油或油质物质是连续相时,称为油包水乳剂。油相可以至少部分地由推进剂例如HFA推进剂组成。油相和水相中的一者或两者均可包含一种或多种表面活性剂、乳化剂、乳液稳定剂、缓冲剂和其他赋形剂。优选的赋形剂包括表面活性剂,尤其是非离子表面活性剂;乳化剂,尤其是乳化蜡;以及液体非挥发性非水材料,特别是二醇,例如丙二醇。油相可含有其他药学上认可的油性赋形剂。例如,如羟化蓖麻油或芝麻油等材料可以在油相中用作表面活性剂或乳化剂。
“洗剂”是低至中等粘度的液体组合物。洗剂可以含有细粉状物质,这些物质通过使用悬浮剂和分散剂而可溶于分散介质中。可替代地,洗剂可以具有与媒介物不混溶的液体物质作为分散相,并且通常借助于乳化剂或其他合适的稳定剂来分散。在一个实施例中,洗剂呈粘度为100至1000厘沱的乳液形式。洗剂的流动性使其可在较宽的表面积上快速均匀地施用。洗剂通常旨在在皮肤上干燥,在皮肤表面留下一层薄薄的药用组分。
“霜剂”是“水包油”或“油包水”型的粘性液体或半固体乳液。霜剂可以含有乳化剂和/或其他稳定剂。在一个实施例中,组合物呈粘度大于1000厘沱(通常在20,000-50,000厘沱的范围内)的霜剂形式。与软膏相比,霜剂通常是在时间上优选的,因为它们通常更易于涂抹和清除。
“软膏”是含有软膏基质和任选地一种或多种活性剂的半固体制剂。合适的软膏基质的实例包括烃类基质(例如矿脂、白矿脂、黄色软膏和矿物油);吸收基质(亲水矿脂、无水羊毛脂、羊毛脂和冷霜);水可去除性基质(例如亲水性软膏)和水溶性基质(例如聚乙二醇软膏)。糊剂通常与软膏不同,因为它们含有较大百分比的固体。与用相同组分制备的软膏相比,糊剂通常具有更高的吸收性和更少的油腻性。
“凝胶”是一种半固体系统,其包含小的或大的分子在液体媒介物中的分散体,该液体媒介物通过溶解或悬浮于该液体媒介物中的增稠剂或聚合物材料的作用呈半固体。该液体可以包括亲脂性组分、水性组分或两者。一些乳剂可以是凝胶或者另外包括凝胶组分。然而,一些凝胶不是乳剂,因为它们不含有不混溶组分的均质共混物。合适的胶凝剂包括但不限于改性纤维素,例如羟丙基纤维素和羟乙基纤维素;卡波普均聚物和共聚物;及其组合。液体媒介物中的合适溶剂包括但不限于二甘醇单乙醚;亚烃基二醇,如丙二醇;二甲基异山梨醇;醇,如异丙醇和乙醇。通常根据溶剂溶解药物的能力来选择溶剂。还可以掺入改善组合物的皮肤感觉和/或润肤性的其他添加剂。此类添加剂的实例包括但不限于肉豆蔻酸异丙酯、乙酸乙酯、C12-C15烷基苯甲酸酯、矿物油、角鲨烷、环甲硅油、癸酸/辛酸甘油三酯或其组合。
还可以使用有助于使皮肤水合的润肤剂或润滑媒介物。“润肤剂”是软化或舒缓皮肤的外部施加试剂,并且通常是本领域已知的且在概略中列出,如“Handbook ofPharmaceutical Excipients[药物赋形剂手册]”,第4版,Pharmaceutical Press[医药出版社],2003。这些包括但不限于杏仁油、蓖麻油、长角豆属提取物、鲸蜡硬脂醇、鲸蜡醇、鲸蜡酯蜡、胆固醇、棉籽油、环甲硅油、乙二醇棕榈硬脂酸酯、甘油、单硬脂酸甘油酯、单油酸甘油酯、肉豆蔻酸异丙酯、棕榈酸异丙酯、羊毛脂、卵磷脂、轻质矿物油、中链甘油三酯、矿物油和羊毛脂醇、矿脂、矿脂和羊毛脂醇、大豆油、淀粉、硬脂醇、葵花油、木糖醇或其组合。在一个实施例中,润肤剂选自由硬脂酸乙基己酯和棕榈酸乙基己酯组成的组。
在特定的实施例中,上述组合物可以配制成气雾剂,在气雾剂中,将该组合物溶于推进剂(例如二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷、二氧化碳或其他合适的气体)以及共溶剂(例如乙醇、丙酮、十六醇等及其组合)中。
在某些实施例中,各种实施例的组合物可以被配制成用于改善皮肤的肌理、外观、颜色、感觉或水合作用,并且其可以另外包括添加剂,例如维生素、美容肽、控油剂和其他皮肤护理剂。
维生素包括例如维生素D、维生素K、维生素B(包括烟酰胺、烟酸、C1-18烟酸酯和烟醇;B6化合物,例如吡哆醇;和B5化合物,例如泛醇或“pro-B5”)、维生素A(包括类视黄醇,例如视黄醇丙酸酯、类胡萝卜素和其他化合物)、维生素E(包括生育酚山梨酸酯、生育酚乙酸酯、生育酚的其他酯)、维生素C(包括脂肪酸的抗坏血酸酯和抗坏血酸衍生物,例如抗坏血酸葡糖苷、抗坏血酸磷酸镁、抗坏血酸磷酸钠和抗坏血酸山梨酸酯)、及其所有天然的和/或合成的类似物、及其组合。在各种实施例中,组合物可包括约0.0001wt.%至约50wt.%、约0.001wt.%至约10wt.%、约0.01wt.%至约5wt.%、或约0.1wt.%至约1wt.%的或任何单个浓度或范围的组合物中所含的每种维生素。
肽包括二肽、三肽、四肽、五肽和六肽,它们的盐、异构体、衍生物及其混合物。有用的肽衍生物的实例包括但不限于衍生自以下的肽:大豆蛋白,棕榈酰基-赖氨酸-苏氨酸(pal-KT)、棕榈酰基-赖氨酸-苏氨酸-苏氨酸-赖氨酸-丝氨酸棕榈酰基-甘氨酸-谷氨酰胺-脯氨酸-精氨酸(这三种可从法国司达玛公司(Sederma)获得),以及Cu-组氨酸-甘氨酸-甘氨酸(Cu-HGG,也称为),及其天然存在和合成的衍生物,及其组合。在各种实施例中,组合物可包括约1x10-7wt.%至约20wt.%、约1x10- 6wt.%至约10wt.%、以及约1x10-5wt.%至约5wt.%的或任何单个浓度或范围的组合物中所含的每种肽。
控油剂包括可用于调节皮肤油或皮脂的产生并改善油性皮肤外观的化合物。控油剂的实例包括例如水杨酸、脱氢乙酸、过氧化苯甲酰、维生素B3(例如烟酰胺)等,它们的异构体、酯、盐和衍生物及其混合物。这样的实施例的组合物可包括约0.0001wt.%至约15wt.%、约0.01wt.%至约10wt.%、约0.1wt.%至约5wt.%、以及约0.2wt.%至约2wt.%的或任何单个浓度或范围的组合物中所含的每种控油剂。
其他皮肤护理剂包括视黄醇、类固醇、防晒剂、水杨酸盐、米诺环素、抗真菌剂、肽、抗体、利多卡因等及其组合。在一些实施例中,其他皮肤护理剂包括N-酰基氨基酸化合物,包括例如N-酰基苯基丙氨酸、N-酰基酪氨酸等,它们的异构体(包括它们的D和L异构体)、盐、衍生物及其混合物。合适的N-酰基氨基酸的实例是N-十一烷基烯酰-L-苯基丙氨酸是以商品名可商购的。其他皮肤活性剂包括但不限于Lavandox、Thallasine 2、Argireline NP、Gatuline In-Tense和Gatuline Expression、Myoxinol LS 9736、Syn-ake和SesaflashTM、N-乙酰基D-葡萄糖胺、泛醇(例如可从阿尔卑斯制药公司(Alps Pharmaceutical Inc.)获得的DL泛醇)、生育酚烟酸酯、过氧化苯甲酰、3-羟基苯甲酸、类黄酮(例如黄烷酮、查尔酮)、法尼醇、植烷三醇、乙醇酸、乳酸、4-羟基苯甲酸、乙酰水杨酸、2-羟基丁酸、2-羟基戊酸、2-羟基己酸、顺式-视黄酸、反式-视黄酸、视黄醇、视黄酯(例如视黄醇丙酸酯)、植酸、N-乙酰基-L-半胱氨酸、硫辛酸、生育酚及其酯(例如生育酚乙酸酯:DL-a-生育酚乙酸酯,可从卫材公司(Eisai)获得)、壬二酸、花生四烯酸、四环素、布洛芬、萘普生、酮洛芬、氢化可的松、对乙酰氨基酚、间苯二酚、苯氧乙醇、苯氧丙醇、苯氧异丙醇、2,4,4'-三氯-2'-羟基二苯醚、3,4,4'-三氯碳酰替苯胺、吡罗克酮乙醇胺盐(octopirox)、利多卡因盐酸盐、克霉唑、咪康唑、酮康唑、新霉素硫酸盐、茶碱、及其混合物。其他皮肤护理剂在美国公开号2007/0020220A1中公开,其中组分/成分通过引用以其整体并入本文。
在一些实施例中,组合物还可包括亮肤剂,例如抗坏血酸化合物、维生素B3化合物、壬二酸、丁基羟基茴香醚、没食子酸及其衍生物、甘草酸、对苯二酚、曲酸、熊果苷、桑树提取物、及其混合物。亮肤剂的组合的使用被认为是有利的,因为它们可以通过不同的机理给皮肤提供增亮益处。
在一些实施例中,组合物包括防晒剂,例如但不限于对氨基苯甲酸(PABA)、PABA酯(甘油基PABA、戊基二甲基PABA和辛基二甲基PABA)、二苯甲酮(氧苯酮和磺异苯酮)、肉桂酸酯(辛基甲氧基肉桂酸酯和西诺沙酯)、水杨酸酯(水杨酸同甲酯)、邻氨基苯甲酸酯、TiO2、阿伏苯宗、贝曲嗪诺(bemotrizinol)、亚甲基双苯并三唑基四甲基丁基苯酚(bisoctrizole)、3-(4-甲基亚苄基)樟脑、西诺沙酯、二乙氨基羟基苯甲酰基己基苯甲酸酯、二羟苯宗、甲酚曲唑三硅氧烷、依莰舒、乙基己基三嗪酮、胡莫柳酯、邻氨基苯甲酸薄荷酯、奥克立林、水杨酸辛酯、二乙基己基丁酰胺基三嗪酮(iscotrizinol)、异戊烯基-4-甲氧基肉桂酸酯、辛基-二甲基-对氨基苯甲酸、辛基-甲氧基肉桂酸酯、氧苯酮、聚硅酮-15、水杨酸三乙醇胺和ZnO。在一些实施例中,本文公开的任何活性剂可以与本文公开的或本领域已知的任何防晒剂组合。
在一些实施例中,组合物还可包括pH调节剂或缓冲剂,其存在于组合物中以中和和/或激活增稠聚合物,以促进具有所需流变特性的组合物的形成。可以使用本领域已知的并且适合用于皮肤接触应用的任何碱或缓冲液系统。在一个实施例中,碱可包括三乙醇胺(例如10%三乙醇胺(TEA)的溶液)、碱金属氢氧化物(例如氢氧化钠(NaOH))、弱酸的盐(例如乳酸铵、柠檬酸钠、抗坏血酸钠)、或其混合物。
在一些实施例中,组合物可以呈水凝胶形式。通常通过使各种单体和/或聚合物交联以提供三维聚合物网络来制备水凝胶。聚合物的非限制性实例包括聚氧乙烯-聚丙烯嵌段共聚物、离子多糖(例如壳聚糖或藻酸钠)、纤维素和可生物降解的聚合物(例如聚丙交酯(PLA)和聚乙交酯(PGA))、琥珀酸丁二酯(PBS)、聚羟基链烷酸酯(PHA)、聚己内酸内酯(PCL)、聚羟基丁酸酯(PHB)、乙醇酸戊酯(PHV)、PHB和PHV共聚物(PHBV)以及聚乳酸(PLA)-聚乙二醇(PEG)共聚物(PLEG))。
在一些实施例中,本文公开的组合物可以呈持续释放组合物的形式。持续(或受控)释放是指活性剂在一段时间内从组合物中逐渐释放。尽管可能存在初始爆发阶段,但在一些实施例中,优选释放表现出相对线性的动力学,从而在释放期内提供恒定的活性剂供应。释放期可能从1到8小时不等,这取决于皮肤障碍及其预期用途。组合物可含有各种可生物降解的聚合物以促进缓慢释放,例如聚丙交酯(PLA)、聚乙交酯(PGA)、聚琥珀酸丁二酯(PBS)、聚羟基链烷酸酯(PHA)、聚己内酸内酯(PCL)、聚羟基丁酸酯(PHB)、乙醇酸戊酯(PHV)、PHB和PHV共聚物(PHBV)以及聚乳酸(PLA)-聚乙二醇(PEG)共聚物(PLEG))。在一些实施例中,优选的聚合物是泊洛沙姆,例如127。
在一些实施例中,组合物可包括纳米颗粒或微粒以进一步促进递送。在一些实施例中,细胞外基质组分、其片段及组合(诱饵分子)和活性剂都可以包封在纳米颗粒和/或微粒内,或者可以与纳米颗粒和/或微粒进行离子缔合。在一些实施例中,仅细胞外基质组分、其片段及组合(诱饵分子)可以被包封在纳米颗粒和/或微粒内。在一些实施例中,仅活性剂可以被包封在纳米颗粒和/或微粒内。在一些实施例中,细胞外基质组分、其片段及组合(诱饵分子)的一部分和/或活性剂可以被包封在纳米颗粒和/或微粒内。在一些实施例中,纳米颗粒或微粒可以是脂质纳米颗粒或脂质微粒。可以通过形成分散或溶解在溶剂中的活性剂、细胞外基质组分或两者的乳液来制备这样的纳米颗粒或微粒,并且该溶液可以与甘油和泊洛沙姆组合以形成乳液。可以将乳液加热、冷却并均质化以产生微粒或纳米颗粒。在一些实施例中,可以商购纳米颗粒或微粒,例如像,杂合聚酰胺基酰胺(PAMAM)树状聚合物水凝胶/聚(乳酸-共-乙醇酸)(PLGA)纳米颗粒或微粒(HDNP)、壳聚糖(CS)纳米颗粒或微粒、硫醇化壳聚糖纳米颗粒或微粒、磷酸钙(CaP)纳米颗粒或微粒、聚(乳酸-共-乙醇酸)共聚物(PLGA)、聚(乙二醇)-嵌段-聚(己内酯)纳米聚合物纳米颗粒或微粒、核/壳纳米颗粒或微粒(例如,由作为核心的卵磷脂脂质体和pluronic F 127二丙烯酸酯(DA-PF 127)构成)、无机物包被的视黄酸(atRA)纳米颗粒或微粒、聚(乳酸)(PLA)均聚物和PEG-嵌段-PLA共聚物纳米颗粒或微粒、PEG-嵌段-PPG共聚物(例如Pluronic)、PEG化脂质体-鱼精蛋白-透明质酸纳米颗粒或微粒、聚乳酸/聚乳酸-聚环氧乙烷(PLA/PLA-PEO)纳米颗粒或微粒等、及其组合。在一些实施例中,纳米颗粒的直径可以为约2至约200纳米、约5至约50纳米、或约18至约22纳米、或这些范围所涵盖的任何范围或单个值。
在一些实施例中,组合物可包括脂质体以促进递送。在一些实施例中,细胞外基质组分、其片段及组合(诱饵分子)和活性剂都可以被包封在脂质体内。在一些实施例中,仅细胞外基质组分、其片段及组合(诱饵分子)可以被包封在脂质体内。在一些实施例中,仅活性剂可以被包封在脂质体内。在一些实施例中,细胞外基质组分、其片段及组合(诱饵分子)的一部分和/或活性剂可以被包封在脂质体内。脂质体是众所周知的并且通常在制药领域中使用,并且在实施例中可以使用任何类型的脂质体。在一些实施例中,脂质体可以是磷脂酰胆碱(PC)和其他成分(例如胆固醇和脂质缀合的亲水性聚合物)。在一些实施例中,脂质体可以是壳聚糖或可以包被在壳聚糖中(即,壳糖体)。在一些实施例中,组合物可以包括胶态脂质。这样的组合物可以包括由以下形成的胶态极性脂质:蓖麻油和/或氢化蓖麻油(例如像PEG-30蓖麻油、PEG-33蓖麻油、PEG-36蓖麻油、PEG-40蓖麻油、PEG-30氢化蓖麻油和PEG-40氢化蓖麻油)的一种或多种非离子聚乙二醇衍生物、阴离子纯化的多糖(例如结冷胶)、一种或多种缓冲剂(例如像硼酸、三甲胺)、以及在一些实施例中一种或多种水性润滑剂和一种或多种胶态水性润滑剂。在一些实施例中,脂质体或胶态脂质可以形成约1纳米至约50纳米、或约6纳米至约22纳米的颗粒。在一些实施例中,组合物可以包括约0.1w/v%至约15w/v%的脂质。
可以使用多种方法制备上述组合物。广义上讲,可通过在足以提供药学上可接受的组合物的温度和时间下将如本文所述的组合物的组分组合在一起来制备组合物。例如,在一些实施例中,组合物的组合物组分能以有效浓度溶解、悬浮、分散或以其他方式混合于所选的载体或赋形剂中,使得缓解或减轻待治疗的病症。
另外的实施例涉及包括上述组合物的装置。例如,可以将此类组合物涂覆在绷带上、与生物粘附剂混合或包括在伤口敷料中。
渗透促进剂
在一些实施例中,组合物可以包括一种或多种用于透皮药物递送的渗透剂或渗透促进剂。化学渗透促进剂(CPE),用于透皮药物递送。CPE是本领域中己知的。合适的CPE可以包括但不限于溶剂(例如一元醇,例如甲醇、乙醇、丙醇、异丙醇)、脂肪酸(例如油酸、辛酸)、脂肪醇(例如月桂醇、肉豆蔻醇、油醇)、表面活性剂(例如离子或非离子洗涤剂)、脂肪酸酯(例如肉豆蔻酸异丙酯、棕榈酸异丙酯、丙酸甲酯和油酸乙酯)、有机酸(例如水杨酸和水杨酸盐、柠檬酸和琥珀酸)、含氮化合物(例如尿素)、胆汁盐和衍生物、胶束/脂质体或形成胶束或脂质体的组分(例如磷脂)、亚砜、萜烯和萜类化合物、多元醇、尿素和衍生物、以及螯合剂。CPE的非限制性列表可包括甲醇、乙醇、丙二醇、乙二醇、甘油、丁二醇、聚乙二醇、聚乙二醇单月桂酸酯、二甘醇、单乙醚(transcutol)、油酸、辛酸、薄荷醇、橙花醇、樟脑、水杨酸甲酯、月桂酸钠、十二烷基硫酸钠、鲸蜡基三甲基溴化铵、苯扎氯铵、泊洛沙姆(231、182、184)、吐温(20、40、60、80)、十二烷基磺酸钠(SDS)、甲基磺酰甲烷(MSM)、苯扎氯铵、聚乙二醇40氢化蓖麻油、二癸基二甲基溴化铵(DDAB)、二癸基三甲基溴化铵(DTAB)、聚山梨醇酯、乙醇酸钠、脱氧胆酸钠、EDTA、柠檬酸、二甲基乙酰胺(DMA)、DMSO、二甲基甲酰胺(DMF)、二甲基亚砜、癸基甲基亚砜、丙二醇、聚乙二醇、甘油、2-吡咯烷酮、1-甲基-2-吡咯烷酮、乙醇胺、二乙醇胺、三乙醇胺、尿素、卵磷脂、萜烯、萜类化合物、1-取代的氮杂环庚烷-2-酮(例如1-正十二烷基环氮杂环庚烷-2-酮)、磷脂、水、及其混合物。适用于本发明的更多渗透促进剂可以是本领域技术人员已知的。(参见例如,Williams等人,“Penetration enhancers[渗透促进剂]”Adv.Drug Deliv.Rev.[高级药物递送评论]2004年3月27;56(5):603-18;以及Pathan等人,“Chemical Penetration Enhancers for Transdermal Drug Delivery System[用于透皮药物递送系统的化学渗透促进剂],”Trop.J.Pharma.Res.[热带药物研究杂志],2009年4月;8(2):173-179;将其内容通过引用以其全文并入本文)。
取决于化合物的类型,渗透剂或渗透促进剂能以约0.1%至约30.0%w/w存在。例如,渗透促进剂能以按重量计约0.1w/w%至约15w/w%的总量包括在实施例的配制品中,并且在各种实施例中,渗透促进剂能以约2w/w%至约12w/w%、约4w/w%至约10w/w%、约4w/w%至约7w/w%、约4w/w%至约6w/w%、约4.5w/w%至约5.5w/w%、约4w/w%至约5w/w%、或这些实例范围内的任何范围或单个浓度的总量包括在内。
在一些实施例中,渗透剂或渗透促进剂可以是不可电离的二醇醚。不可电离的二醇醚包括例如二乙二醇单甲醚、三乙二醇单甲醚、聚乙二醇单甲醚、乙二醇单乙醚、二乙二醇单乙醚、三乙二醇单乙醚、乙二醇单异丙醚、乙二醇单丁醚、二乙二醇单丁醚、三乙二醇单丁醚、乙二醇单异丁醚、二乙二醇单己醚、乙二醇单2-乙基己醚、二乙二醇单2-乙基己醚、乙二醇单烯丙基醚、乙二醇单苯基醚、乙二醇单苄基醚、二乙二醇单苄基醚、丙二醇单甲醚、二丙二醇单甲醚、三丙二醇单甲醚、二丙二醇单丙醚、丙二醇单丁醚、二丙二醇单丁醚、丙二醇单苯基醚、乙二醇二甲醚、二乙二醇二甲醚、三乙二醇二甲醚、二乙二醇二乙醚、二乙二醇二丁醚、二丙二醇二甲醚等及其组合。这些不可电离的二醇醚的二醇部分包括多种化学品,包括但不限于丙二醇、二丙二醇、丁二醇、以及通式为HOCH2(CH2OH)nCH2OH(其中n(氧乙烯基团数)是4-200)的聚乙二醇。在特定的实施例中,不可电离的二醇醚可以是二乙二醇单乙醚(“DEGEE”或“乙氧基二甘醇”,以其商品名已知,可从新泽西州帕拉默斯市的嘉法狮公司(Gattefosse,Paramus,N.J)商购)。
不可电离的二醇醚能以约0.01w/w%至50w/w%的浓度包含在各种实施例的组合物中,并且在特定的实施例中,不可电离的二醇醚的浓度可以为约0.5w/w%至约10w/w%。在各种实施例中,不可电离的二醇醚能以约1w/w%至约25w/w%、约2w/w%至约20w/w%、约4w/w%至约10w/w%、约4w/w%至约8w/w%、约4w/w%至约5w/w%、或这些实例范围内的任何范围或单个浓度的总量包括在内。
在一些实施例中,渗透剂或渗透促进剂可以是肽或蛋白质片段。这样的肽和蛋白质片段通常被称为“皮肤穿透肽”(SPP)或细胞穿透肽(CPP)。SPP可以稳定皮肤中的这些结构蛋白,而不会使它们变性。例如,SPP通过氢键和弱的静电相互作用与角蛋白结合,并可作为角蛋白和药物分子之间的结合介质起作用。SPP还可以通过细胞之间的间隙以及通过脂双层使药物不受破坏地扩散而利用角质细胞之间的途径。SPP的一个实例是TD-1,已知它可以使角质细胞之间的桥粒诱导的紧密连接松弛,并在局部应用后30分钟内使细胞之间的距离从30nm变为466nm。用TD-1处理后1小时,细胞间隙增加,然后逐渐恢复。各种SPP是本领域已知的,并且已显示出许多包含9至19个氨基酸的肽表现出皮肤渗透活性。实施例涵盖所有此类肽。
在一些实施例中,渗透促进剂和诱饵分子均有助于活性剂渗透穿过表面组织。在一些实施例中,渗透促进剂和诱饵分子以协同方式共同作用。在一些实施例中,渗透促进剂特别有助于疏水活性剂的渗透。
在一些实施例中,组合物缺乏渗透促进剂。
透皮贴剂
在一些实施例中,本文公开的组合物可以呈透皮贴剂的形式。透皮贴剂可以呈任何常规形式,例如像,条带、纱布、薄膜等。贴剂材料可以是非织造的或织造的(例如,纱布敷料)。在加工过程中也可以层压各层。它可以是非闭合的或闭合的,但是后者对于衬层是优选的。优选将贴剂气密密封以用于存储(例如,箔包装)。贴剂可以保持在皮肤上,并且贴剂的组分可以使用各种粘附剂保持在一起。例如,透皮贴剂可以是创可贴型装置的形式,或者其可以包装在小的金属或塑料“杯子”中,使用粘附剂、胶带或外部织物或皮带将其绑在适当的部位上,类似于手表的一部分。整个贴剂可以是一次性的或可以是可再填充的。
本文公开的透皮贴剂可以由任何聚合材料制成。可以用于透皮贴剂中的聚合物基质材料的非限制性实例是乙烯乙酸乙烯酯(EVA)共聚物、交联聚(乙烯醇)、聚(羟基甲基丙烯酸乙酯)、酰基取代的醋酸纤维素、水解的烯烃-乙酸乙烯酯共聚物、聚氯乙烯、聚醋酸乙烯酯、聚乙烯烷基醚、聚氟乙烯、聚碳酸酯、聚氨酯、聚酰胺、聚砜、苯乙烯丙烯腈共聚物、交联聚(环氧乙烷)、聚(烯烃)、聚(乙烯基咪唑)、聚(酯)、聚(对苯二甲酸乙二醇酯)、聚磷腈、氯磺化聚烯烃、聚丙交酯(PLA)、聚乙交酯(PGA)、或其组合。
在一些实施例中,透皮贴剂可以呈水凝胶的形式。在一些实施例中,呈透皮贴剂的组合物可进一步包括以下组分,例如粘合剂(binder)、缓冲剂、着色剂、干燥剂、稀释剂、湿润剂、防腐剂、稳定剂、其他赋形剂、粘附剂(adhesive)、增塑剂、增粘剂、增稠剂、冷却剂、或其组合。
使用组合物的方法
本文公开了治疗各种障碍的方法。将包含本文所述的活性剂、透明质酸酶/弹性蛋白酶和诱饵分子的组合物施用于表面组织。施用通常是局部、透皮、经皮或微针注射。“表面组织”包括任何组织表面,例如但不限于皮肤、粘膜、眼睛、耳朵、鼻子内部、口腔内部、嘴唇、尿道开口、阴道、肛门、舌头、舌系带、毛发、牙齿、骨骼、泪腺、鼻窦粘膜、呼吸道、牙龈等。在一些实施例中,表面组织是皮肤表面或粘膜表面。在一些实施例中,粘膜表面可以是眼睛。在一些实施例中,粘膜表面可以是口腔或阴道腔。
在一些实施例中,所施用的组合物包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,所施用的组合物包含一种或多种活性剂和选自透明质酸酶、弹性蛋白酶或其组合的酶。在一些实施例中,所施用的组合物包含一种或多种活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子,并且进一步包含选自透明质酸酶、弹性蛋白酶或其组合的酶。在一些实施例中,所施用的组合物包含一种或多种活性剂,并且不包含诱饵分子或透明质酸酶/弹性蛋白酶。在一些实施例中,所施用的组合物仅包含诱饵分子而不包含活性剂。在一些实施例中,所施用的组合物包含选自透明质酸酶、弹性蛋白酶或其组合的酶,并且不包含活性剂。
皮肤障碍
本发明的实施例涉及在有需要的受试者中治疗皮肤病症的方法,这些方法包括向该受试者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,这些组合物包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。在实施例中,皮肤病症选自痤疮、银屑病、特应性皮炎、酒渣鼻及其组合。
在一些实施例中,皮肤病症可以是良性肿瘤、初癌或恶性肿瘤。在一些实施例中,皮肤病症可以选自人乳头瘤病毒诱导的病变,例如疣、普通疣、掌跖疣、扁平疣、疣状表皮发育不良相关疣、肛殖疣、尖锐湿疣;疱疹病毒相关病变,包括由HHV-1(HSV-1)、HHV-2(HSV-2)、HHV-3(水痘带状疱疹病毒)引起的病变,例如水痘、带状疱疹(Herpes zoster)、带状疱疹(shingles);痘病毒引起的病变,例如传染性软疣、羊痘疮;胼胝、皮角、鸡眼、软垂疣、纤维上皮息肉、结节性痒疹、日光性角化症、鳞状细胞癌、原位鳞状细胞癌、角化棘皮瘤、基底细胞癌、皮肤淋巴瘤和良性淋巴细胞性浸润与皮肤增生、透明细胞棘皮瘤、大细胞棘皮瘤、表皮松解性棘皮瘤、汗孔角化病、角化过度、苔癣样角化病、棘皮症、黑棘皮症、融合性网状乳头瘤病、痣(包括例如真皮痣、表皮痣、复合痣、ILVEN(炎性线状疣状表皮痣)、皮脂腺痣、黑头粉刺样痣等);痤疮,例如粉刺型痤疮、炎性痤疮、丘疹性痤疮、脓疱性痤疮、囊性痤疮;囊肿,例如表皮样囊肿、粟丘疹、毛膜囊肿、滤泡囊肿、增生性囊肿、皮样囊肿、藏毛囊肿、大汗腺性囊肿、外分泌腺性囊肿、皮脂囊肿、粘液囊肿、粘液样囊肿、腱鞘囊肿、滑液囊肿、毳毛囊肿、皮脂腺囊瘤、汗腺囊瘤;附件肿瘤,例如毛囊瘤、纤维毛囊瘤、毛囊周围纤维瘤、毛盘瘤、皮脂腺痣、软骨样汗管瘤、毛上皮瘤、毛母细胞瘤、促结缔组织增生性毛上皮瘤、毛母质瘤、毛母质癌、毛根鞘肿(tricholemmoma)、外毛根鞘癌、毛囊漏斗肿瘤、毛发腺瘤、增殖性毛发肿瘤、皮脂腺增生、皮脂腺腺瘤、皮脂腺上皮瘤、皮脂腺癌、汗管瘤、汗孔瘤、汗腺腺瘤、顶泌汗腺腺瘤、螺旋腺瘤、圆柱瘤、小汗腺痣(小汗腺错构瘤)、乳头状腺瘤、乳头状腺癌;良性黑素细胞瘤,例如雀斑、牛奶咖啡色斑(café-au-lait macule)、贝克尔黑变病、雀斑样痣、日光性斑痣、单纯性雀斑样痣、粘膜黑素细胞病变、胎斑、太田痣、蓝色痣、常见的获得性黑素细胞痣(痣细胞痣,“痣(mole)”)、先天性痣、斑痣、复发性痣;血管和血管周围肿瘤和反应性增生,例如血管瘤、樱桃状血管瘤、钉状血管瘤(靶向性血铁质性血管瘤)、簇状血管瘤、血管内皮瘤、伴有嗜酸性粒细胞增多症的血管淋巴样增生(ALHE)、血管球瘤(glomus tumor/glomangioma)、血管外皮瘤;皮肤神经瘤和神经内分泌瘤,例如神经瘤、神经鞘瘤、神经纤维瘤、神经鞘膜肿瘤、神经鞘粘液瘤(nerve sheath myxoma)、神经鞘瘤(neurothekeoma)、颗粒细胞瘤;纤维化增生和纤维组织细胞增生,例如软垂疣、纤维上皮息肉、纤维瘤、纤维性丘疹、血管纤维瘤、珍珠状阴茎丘疹、甲周纤维瘤、皮肤纤维瘤、纤维角质瘤、硬化型纤维瘤或多形性纤维瘤、结缔组织痣;皮肤疤痕、增生、瘢痕疙瘩、酒渣鼻、皮肤真菌、皮肤癣菌和霉菌感染、甲癣、色素沉着过度、皱纹、银屑病、恶性黑色素瘤、脂溢性角化病、脂溢性角化病变异(包括例如黑质丘疹皮肤病、倒转毛囊角化病/角质瘤疣状角化不良病/疣状角化不良瘤、疣状肢端角化病、灰泥样角化病);或其组合。
本文公开的组合物可包括治疗皮肤病症的任何活性剂。活性剂的非限制性实例包括水杨酸、氢氧化钾、鬼臼脂、斑蝥素、咪喹莫特、硝酸、口服西咪替丁、5-氟尿嘧啶、博来霉素、DNCB、咪喹莫特和三氯乙酸、过氧化苯甲酰、博来霉素、2,4-二硝基氯苯、氟尿嘧啶、水杨酸、硝酸银、硫酸锌、氧化锌、盐酸克林霉素和磷酸克林霉素、红霉素、四环素、双氯青霉素、强力霉素、米诺环素、杆菌肽、金霉素、新霉素、莫匹罗星、多粘菌素B、铜迈星、呋喃唑酮、庆大霉素、林可霉素、头孢菌素、β内酰胺抗生素、盐酸可可霉素、他扎罗汀、维生素A、视黄酸、维甲酸、异维甲酸、阿达帕林、视黄醇、阿维A、蓓萨罗丁、类视黄醇;奥昔布宁;维生素D、维生素C、维生素B、维生素E;硫;糖皮质激素、皮质类固醇、曲安西龙、曲安奈德、倍他米松、倍他米松1 7-戊酸酯、倍他米松二丙酸酯、哈西奈德、醋酸异氟泼尼松、氟米松、氟轻松、莫米松、氟替卡松、丙酸氟替卡松、泼尼松龙、倍氯米松、氢化可的松、环丙孕酮、屈螺酮、雌激素、孕激素、他克莫司、吡美莫司、熊果酸、桦木酸、模绕酮酸(moronic acid)、齐墩果酸、阿昔洛韦、伐昔洛韦、泛昔洛韦、喷昔洛韦、二十二醇、紫苏醇、及其组合。
本发明的实施例涉及在有需要的受试者中改善皮肤肌理的方法,这些方法包括向该受试者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。在一些实施例中,改善皮肤肌理选自改善光度、质量及其组合。
在一些实施例中,一种治疗皮肤表面上的疤痕和皱纹的方法包括向皮肤表面施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。
本文公开的治疗性组合物可具有美容目的。该组合物可例如用于改善皮肤外观,例如通过减少或去除脸部线条和皱纹、以及减少或去除妊娠纹。根据一个实施例,一种改善皮肤外观的方法包括以足以刺激弹性发生的量施用该组合物。此外,本文公开的组合物可收紧脸部和身体其他部分(包括手臂、腿、胸部和颈部区域)上松弛、下垂的皮肤,或给出减少皱纹的外观。使用组合物的其他方法包括刺激平滑肌细胞和牙龈成纤维细胞以分别产生弹性蛋白和原纤蛋白(耐酸纤维),分别用于治疗新生内膜增厚和牙齿松动(牙龈炎)。根据一些实施例,本公开还考虑了用于刺激真皮细胞分化的组合物和方法。
此外,组合物可用于增强伤口愈合并预防和治疗皮肤增生性瘢痕。因此,本公开的另一个实施例包括一种促进伤口愈合和减少瘢痕化的方法,该方法包括将预处理的成纤维细胞组合物以足以刺激弹性蛋白在受伤部位沉积的量施加至伤口。在一些实施例中,包含产生弹性蛋白或胶原蛋白的治疗性细胞的组合物可用于促进伤口愈合或治疗疤痕。适用于治疗的伤口包括那些由外伤(例如烧伤、擦伤和割伤)引起的伤口、由外科手术(例如切口和皮肤移植)引起的伤口、皮肤伤口、角膜伤口和溃疡。受伤部位的弹性蛋白合成也可以减少疤痕化,因为疤痕组织不含弹性蛋白,弹性蛋白是未受伤皮肤的重要组分。对弹性蛋白的刺激和其分泌到基质中通常还可以为参与愈合过程的细胞提供有利的环境,从而进一步促进伤口愈合过程。
毛发障碍
本文公开的各种实施例的组合物可用于治疗毛发、毛干、毛囊、毛球、油腺、及其组分。因此,某些实施例包括通过使毛发、头皮、或含有毛发或毛囊的皮肤与上述组合物接触来施用活性剂的方法。本文所述的组合物和方法可用于向毛发、毛干、毛囊、毛球、油腺、及其成分施用任何活性剂,包括小分子药物、大分子药物、生物制剂、抗体、嵌合抗体、肽、抗氧化剂等及其组合。
各种实施例的组合物可以包括用于治疗与毛发、毛干、毛囊、毛球、油腺、及其组分有关的疾病的任何治疗剂,这些疾病包括例如脱发、头皮屑、脂溢性皮炎、斑秃、毛发病、癣、头癣、毛囊炎、模式脱发、休止期脱发、乳痂、拔毛癖、牵引性脱发、结节性脆发症、脱发性毛囊炎、头虱感染、前额纤维化脱发、非瘢痕性脱发、石棉状糠疹、头皮切割性蜂窝组织炎、颈项部瘢痕性痤疮、念珠状发、虱病、全秃、假斑状秃发、泡沫毛发畸形、毛发管型、多毛症、毛发内生、念珠状发、头发过早变白、模式脱发、套叠性脆发症等。这些试剂可被设计用于局部(topical)、全身或局部(local)递送,并且活性剂的非限制性实例包括生物制剂、治疗性肽、仿生肽、小分子和大分子镇痛剂、类固醇、以及染料和着色剂。特定实施例中,活性剂可以是但不限于米诺地尔(Rogaine)、非那雄胺(Propecia)、螺内酯(Aldactone)、西咪替丁(Tagmet)、曲安西龙(Kenalog、Triderm)、强的松、倍他米松(Diprolene)、氟轻松(Synalar-HP)、氯倍他索(Clobex、Cormax、Embeline、Temovate)、地蒽酚(Dritho-Creme、Dritho-Scalp、Zithranol-RR)、磺胺嘧啶(sulfazine)、柳氮磺吡啶(Azulfidine)、环孢菌素(Gengraf、Neoral、Restasis、Sandimmune)等及其组合。活性剂的其他实例包括染料和染发剂和/或湿润剂、润肤剂、油、蛋白质、硅酮、表面活性剂、聚合物或其他毛发调理剂。各种实施例的组合物可包括以上列出的任何一种或多种活性剂。例如,一些实施例的组合物可包括毛发生长剂、着色剂或染料、以及调理剂,并且在其他实施例中,组合物可包括生长剂、染料或调理剂中的一种。
上述各种实施例的组合物可用于将足够的活性剂递送至毛发、毛干、毛囊、毛球、或油腺以影响治疗。在特定的实施例中,诱饵可使活性剂穿透表皮和真皮至毛囊和毛球。例如,某些活性剂(例如米诺地尔)不能很好地穿透皮肤层,从而限制了在没有化学载体(例如二甲基亚砜DMSO)或机械递送装置(例如微针)的情况下有效接触毛囊的活性剂的量。包括活性剂和诱饵的组合物可以提供足够的渗透和活性剂向毛囊的递送,而无需使用化学载体或机械递送系统,从而减少潜在的副作用和/或对表皮和头皮的伤害。另外,上述诱饵使活性剂穿透毛干,从而增加了活性剂、染料、毛发调理剂等向毛干的递送。例如,使用包含带有诱饵的染发剂的组合物可以实现改善的染料渗透、着色和保色。类似地,毛发调理剂(例如湿润剂和蛋白质)可以与诱饵组合以产生将此类调理剂递送至毛干的组合物。
其他障碍
本发明的实施例涉及在有需要的受试者中改善运动任务和/或功能性能的执行的方法,这些方法包括向该受试者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。在实施例中,由于例如极端或过高的温度(温暖或寒冷),改善有需要的受试者的运动任务和/或功能性能的执行包括改善受试者的精细运动技能。
本发明的实施例涉及在有需要的受试者中治疗雷诺氏综合症的方法,这些方法包括向该患者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。
本发明的实施例涉及在有需要的受试者中治疗跛行的方法,这些方法包括向该患者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。
本发明的实施例涉及在有需要的受试者中治疗周围血管疾病的体征和症状的方法,这些方法包括向该受试者的表面组织局部施用本文所述的组合物。该组合物可以包含有效量的如本文所述的活性剂和选自细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,该组合物可以进一步包含透明质酸酶/弹性蛋白酶。在一些实施例中,组合物可包含活性剂和透明质酸酶/弹性蛋白酶,并且不包含诱饵分子。
在一些实施例中,本文公开的组合物可用于治疗各种病症,例如但不限于慢性疼痛、术后疼痛、尿失禁、神经障碍(阿尔茨海默氏病、痴呆、帕金森氏病、下肢不宁综合征、抑郁症、神经性疼痛、精神分裂症、睡眠障碍、认知障碍)、心绞痛、冠心病、COPD、恶心、晕动病、避孕、激素治疗、关节炎、骨关节炎、类风湿性关节炎、炎症性肠病、成瘾、ADHD、抗炎性病症、皮肤障碍、乳腺癌、勃起功能障碍、维生素缺乏症、钙缺乏症、糖尿病、糖尿病性神经病、糖尿病足、绝经后症状、潮热、激素替代疗法、偏头痛、疱疹感染、牙龈炎症、肾衰竭、耳鸣、网球肘、肌腱炎、脂解、腕管综合征、性腺机能减退、无血管性坏死、引产、周围神经性疼痛、脊髓损伤、口腔粘膜炎和高血压。
在一些实施例中,本文公开的组合物可用于治疗与甲有关的病症,例如博氏(Beau's)线、甲松离、甲癣、甲脱离、甲沟炎、甲内生、霉菌病、黄甲综合征、脆甲症、凹甲、甲下血肿、白甲病、银屑病性甲营养不良、点刺甲(stipple nail)、甲弯曲等。
在一些实施例中,本文公开的组合物可用于治疗与眼睛有关的病症,例如弱视、睑缘炎、睑板腺囊肿、结膜炎、角膜擦伤、干眼症、糖尿病性视网膜病、青光眼、角膜炎、麦粒肿(hordeolum)、葡萄膜炎、睑腺炎(sty)等。
在一些实施例中,本文公开的组合物可以用于治疗与颊腔或口腔有关的病症,例如口腔癌、鹅口疮、溃疡、牙龈炎、疮、粘膜白斑病、吸烟者上颚、口腔念珠菌病、细菌和病毒感染等。
在一些实施例中,本文公开的组合物可用于治疗与鼻腔有关的病症,例如鼻炎、鼻息肉、鼻窦感染、上呼吸道感染等。
在一些实施例中,本文公开的组合物可用于治疗阴道疾病,例如阴道炎、阴道分泌物、淋病、细菌性阴道病、性传播疾病、萎缩性阴道炎、酵母菌感染、生殖器疣、阴道癌等。
在一些实施例中,本文公开的组合物可用于治疗与肛门有关的病症,例如痔疮、肛门癌、肛门瘙痒症、肛瘘等。
在一些实施例中,本文公开的组合物可用于治疗与舌有关的病症,例如正中菱形舌炎、萎缩性舌炎、裂纹舌、地图样舌、口腔毛状白斑、扁平苔藓、白线、鳞状细胞癌、乳头状瘤、巨舌等。
在一些实施例中,本文公开的组合物可用于治疗鼻窦病症,例如鼻窦炎、鼻鼻窦炎、急性鼻窦炎、亚急性鼻窦炎、亚急性鼻鼻窦炎、慢性鼻窦炎、慢性鼻鼻窦炎、慢性鼻鼻窦炎急性加重、真菌性鼻窦疾病、伴息肉的鼻窦炎、鼻窦肿瘤等。
在一些实施例中,本文公开的组合物可用于治疗呼吸道病症,例如肺癌、间质性肺病、肺栓塞、慢性阻塞性肺病、肺炎、气胸、肺动脉高压、胸腔积液、非小细胞肺癌、哮喘、肺纤维化、阻塞性肺病、呼吸系统疾病、结节病、支气管炎、肺结核、特发性肺纤维化、囊性纤维化、牵引性支气管扩张、肺炎、呼吸衰竭、细支气管炎、过敏性肺炎、限制性肺病、寻常型间质性肺炎、肺部感染、急性呼吸窘迫综合征、胸膜炎、尘肺、煤工尘肺、低氧血症、皮肌炎、灼热性胸痛、肺孢子菌肺炎等。
在一些实施例中,本文公开的组合物可用于治疗与泪管相关的病症,例如鼻泪管阻塞、部分泪管阻塞、总泪管阻塞、泪管狭窄、泪腺炎、泪囊炎、先天性泪囊炎、眼部感染等。
在一些实施例中,本文公开的组合物可用于治疗与内耳相关的病症,例如内耳感染、梅尼埃病、眩晕、自身免疫性内耳疾病、噪声性耳聋、听神经瘤、良性阵发性位置性眩晕、药物引起的耳毒性、耳带状疱疹、化脓性迷路炎、前庭神经元炎等。
在一些实施例中,本文公开的组合物可用于治疗膀胱病症,例如膀胱癌、尿路感染、膀胱膨出、间质性膀胱炎、膀胱过度活动症、尿失禁、膀胱疾病、尿潴留、良性前列腺增生、神经原性膀胱功能障碍、膀胱输尿管反流、压力性尿失禁、胃肠道疾病、夜遗尿等。
在一些实施例中,本文公开的组合物可用于治疗肾脏病症,例如急性肾衰竭或损伤、肾盂肾炎、慢性肾脏疾病、多囊性肾病、肾脏疾病、肾小球肾炎、肾脏疼痛或结石、红斑狼疮、肾病综合征、肾炎、糖尿病性肾病、IgA肾病、常染色体显性多囊性肾脏疾病、囊性肾脏疾病、肾囊肿、奥尔波特综合征、肾小管性酸中毒、古德帕斯丘综合征、髓质海绵肾等。
在一些实施例中,本文公开的组合物可用于治疗尿道病症,例如肾盂积水,菌尿症,血尿症,细菌、真菌和酵母菌感染等。
这些实施例的方法可用于治疗几乎任何病症。例如,实施例的方法可用于治疗各种皮肤病症,包括痤疮、局部疼痛缓解、局部真菌或细菌感染、皮肤癌、脓肿、蜂窝组织炎、肌理、外观、感觉、水合作用等。在其他实施例中,这些方法可用于施用各种美容治疗剂,例如,用于改善皮肤厚度、弹性、复原力、光滑度、肤色、肌理、亮度、透明度、轮廓、硬度、紧致度、柔软度、变色、皮肤损害等及其组合。在其他实施例中,这些方法可以与各种美容治疗剂结合使用。其他实施例的方法可用于增强例如毛发或牙齿的颜色或强度。在仍其他实施例中,本发明的方法可用于施用活性剂以治疗其中优选透皮递送活性剂的多种全身性病症,例如慢性疼痛缓解、癌症、晕动病、慢性疾病等及其组合。
本发明的实施例涉及在有需要的受试者中治疗、减少或改善抬头纹(例如,眉间纹)、皱纹或鱼尾纹的外观的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的如本文所述的活性剂和细胞外基质组分、其片段及其组合。在一些实施例中,活性剂是神经毒素。在一些实施例中,抬头纹、皱纹或鱼尾纹可以是细的、中度的或严重的或其组合。
本发明的实施例涉及在有需要的受试者中治疗出汗过多症状的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的如本文所述的活性剂和细胞外基质组分、其片段及其组合。在一些实施例中,活性剂是神经毒素。在一些实施例中,出汗是严重的腋窝出汗(严重的原发性腋窝多汗症)。在一些实施例中,出汗是腋窝出汗、手出汗、足部出汗、自我感知的或外部感知的过度出汗及其组合。在一些实施例中,受试者为18岁及以上。在一些实施例中,受试者对皮肤上使用的其他药物(局部)没有反应。
本发明的实施例涉及在有需要的受试者中治疗偏头痛的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的如本文所述的活性剂和细胞外基质组分、其片段及其组合。在一些实施例中,活性剂是神经毒素。在实施例中,偏头痛可以是阵发性偏头痛。在一些实施例中,偏头痛可以是慢性偏头痛。
本发明的实施例涉及在有需要的受试者中治疗病症的方法,这些方法包括向该受试者的表面组织局部施用组合物,该组合物包含有效量的如本文所述的活性剂和细胞外基质组分、其片段及其组合。在实施例中,该病症可以选自膀胱过度活动症尿失禁,以预防带有慢性偏头痛的头痛、增加的肌肉僵硬、异常头位和伴随颈部肌张力障碍(CD)发生的颈部疼痛、斜视或眼睑痉挛。在实施例中,该病症可以是皮肤病症。在实施例中,该病症可以是雷诺氏综合征、出汗过多、皮肤肌理改善、跛行或周围血管疾病。在实施例中,该病症可以选自膀胱过度活动症尿失禁、以预防带有慢性偏头痛的头痛、增加的肌肉僵硬、异常头位和伴随颈部肌张力障碍(CD)发生的颈部疼痛、斜视或眼睑痉挛、头痛、勃起功能障碍、抑郁症、足底肌膜炎、皮肤肌理和外观、出汗、多汗症、以及以下外观的暂时改善:(i)与皱眉肌和/或降眉间肌肌肉活动相关的中度至重度眉间纹,(ii)与眼轮匝肌活动相关的中度至重度外侧眼角纹,或(iii)与额肌肌肉活动相关的中度至重度前额纹。在实施例中,该病症可以选自:当另一种类型的药物(抗胆碱能药物)效果不佳或无法服用时,18岁及以上的成年人的膀胱过度活动症(例如,强烈需要漏泄或润湿事故的小便(急迫性尿失禁)、强烈需要立即小便(尿急)和经常小便(尿频));治疗因神经疾病导致膀胱过度活动症的18岁及以上成年人(他们在尝试抗胆碱药物后仍存在漏尿或不能忍受副作用)的漏尿(失禁);预防患有慢性偏头痛的成年人的头痛,他们在18岁或以上的人群中每月有15天或更长时间的头痛,每天头痛持续4个小时或更长时间;治疗患有上肢和下肢痉挛的18岁及以上的人的肘部、腕部、手指、拇指、脚踝和脚趾肌肉的肌肉僵硬增加;或治疗16岁及以上的人的异常头位和伴随颈部肌张力障碍(CD)发生的颈部疼痛;治疗12岁及以上的人的某些类型的眼肌问题(斜视)或眼睑异常痉挛(眼睑痉挛)、勃起功能障碍、抑郁症、足底肌膜炎、皮肤肌理和外观、在对抗胆碱能药物反应不足或不耐受的成年人中的伴有急迫性尿失禁、尿急和尿频症状的膀胱过度活动症(OAB)、在对抗胆碱能药物反应不足或不耐受的成年人中的由于与神经病症(例如脊髓损伤(SCI)、多发性硬化症(MS))相关的逼尿肌过度活动导致的尿失禁;预防患有慢性偏头痛(每月≥15天,每天头痛持续4小时或更长时间)的成年患者的头痛,成年患者的痉挛,成年患者中局部用药不足的严重腋窝多汗症,≥12岁患者的与肌张力障碍相关的眼睑痉挛,以及以下外观的暂时改善:(i)与皱眉肌和/或降眉间肌肌肉活动相关的中度至重度眉间纹,(ii)与眼轮匝肌活动相关的中度至重度外侧眼角纹,或(iii)与额肌肌肉活动相关的中度至重度前额纹。
癌症治疗
在一些实施例中,一种在受试者中引发免疫应答的方法包括向组织表面施用组合物,该组合物包含多个表达抗原的治疗性细胞和选自如本文所述的细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,抗原可以是肿瘤抗原。肿瘤抗原包括但不限于胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤。
在一些实施例中,一种治疗受试者中的癌症的方法包括向组织表面施用组合物,该组合物包含多个表达抗原的治疗性细胞和选自如本文所述的细胞外基质组分、其片段及其组合的诱饵分子。在一些实施例中,抗原可以是肿瘤抗原。肿瘤抗原包括但不限于胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤。
在一些实施例中,本文公开的方法可以与任何过继细胞转移(ACT)疗法组合使用。ACT是免疫疗法的非常有效的形式,并且其涉及将具有抗肿瘤活性的免疫细胞转移到癌症患者体内。ACT涉及在体外鉴定具有抗肿瘤活性的淋巴细胞,将这些细胞体外扩增至大量,并将其输注到携带癌的宿主中。用于过继转移的淋巴细胞可来源于切除的肿瘤的基质(肿瘤浸润性淋巴细胞或TIL)。如果它们经过基因工程化以表达抗肿瘤T细胞受体(TCR)或嵌合抗原受体(CAR)(富含混合淋巴细胞肿瘤细胞培养物(MLTC)或使用自体抗原呈递细胞和肿瘤衍生肽进行克隆),则它们也可以源自或来自血液。其中淋巴细胞起源于要被输注的携带癌的宿主的ACT称为自体ACT。
非侵入性局部诊断
本文公开的组合物可用于非侵入性局部诊断和成像。在一些实施例中,该方法包括施用透皮贴剂,该透皮贴剂包含组合物,该组合物包括一种或多种活性剂和选自如本文所述的细胞外基质组分、其片段及其组合的诱饵分子诱饵分子。在一些实施例中,该组合物可以包含透明质酸酶/弹性蛋白酶代替诱饵分子。据信,诱饵分子或透明质酸酶/弹性蛋白酶会暂时破坏细胞-细胞(即,细胞间)和细胞-支架的附着,从而使间质液、汗液、淋巴液、血清、血液和其他体液扩散并到达组织表面,例如皮肤表面。扩散的液体被透皮贴剂吸收或收集,并对液体进行分析物、抗原、pH、代谢物、电解质等分析。此类方法可以用于非侵入性地收集并诊断体液。
在一些实施例中,透皮贴剂包括液体吸收垫。液体吸收垫可以由任何方便的材料制成,其中合适的目的材料包括但不限于纤维素材料、聚合物材料等。在某些情况下,吸收垫材料由亲水性材料制成。在其他实施例中,吸收贴剂选自由纱布、纤维素垫、琼脂糖凝胶、丙烯酰胺凝胶组成的组。在一些实施例中,透皮贴剂呈本文公开的水凝胶形式。
在一些实施例中,吸收垫在其上表面被闭合层覆盖,下表面涂覆有粘合剂以将该垫固定到皮肤上。公开的垫是一种有点吸收性的材料,能够用作储器,并且由微孔聚酯或聚醚多孔聚氨酯泡沫层、棉、无纺布或类似的能够保留但仍可分配液体载体的布样材料形成。
可以检测到的分析物包括但不限于重金属,例如铅、镉、锂、铜、铁和汞;有机化合物;生物分析物和/或代谢物,例如氧气、葡萄糖、乳酸盐、半乳糖、乙醇、谷氨酸盐、果糖、肌酸、肌酸酐、胆红素、尿素、尿酸、白蛋白;电解质,例如氨、钙、二氧化碳、氯化物、锂、镁、磷、钾、钠;毒素;脂质,例如胆固醇、甘油三酯;激素,例如胰岛素、11-脱氧皮质醇、17-羟孕酮、雄烯二酮、硫酸DHEA、二聚抑制素A、雌二醇、FSH、hCG、hCG、黄体化激素(LH)、PAPP-A、孕酮、催乳素、SHBG、睾酮;治疗剂和药理剂;滥用药物,例如可卡因、咖啡因、酒精和对乙酰氨基酚;娱乐性药物;氨基酸;血液气体;酶,例如胰酶和肝酶;抗生素;细胞因子;蛋白质;生物标志物;心脏标志物;炎性疾病标志物,例如C反应蛋白;肿瘤标志物;细菌抗原和病毒抗原;以及其他生物学相关分子。
可以检测到的分析物的其他非限制性实例包括柠檬酸铁铵、蜂蜡、洋地黄皂苷、对氨基苯乙酮、二氯醌氯亚胺、二氯苯酚和丁酸。
提取之后,可以通过任何标准的化学、物理、酶学和/或光学手段进行分析物的检测和定量。然后使用电化学、生物化学、光学、荧光、吸光度、反射率、拉曼、磁性、质谱、IR光谱测量方法及其组合来检测副产物。
在一些实施例中,本文公开的方法可用于非侵入性地检测抗原。在一些实施例中,该方法涉及施用透皮贴剂,该透皮贴剂包含组合物,该组合物包括抗体或其片段和选自如本文所述的细胞外基质组分、其片段及其组合的诱饵分子诱饵分子。在一些实施例中,该组合物可以包含透明质酸酶/弹性蛋白酶代替诱饵分子。诱饵分子或透明质酸酶/弹性蛋白酶的存在通过暂时破坏细胞-细胞(即,细胞间)和细胞-支架的附着而引起组合物接触的组织发生重排,从而使抗体和抗体片段非侵入性地渗透并穿过组织并结合抗原。结合的抗体可以通过成像进行检测。抗体或抗体片段可以是本文所述的任何抗体,例如但不限于识别肿瘤抗原、癌症抗原、过敏原、细菌抗原、病毒抗原、药物、激素、植物凝集素、内毒素及其组合的抗体及其片段。
透皮贴剂可以应用于任何组织表面,例如但不限于皮肤、粘膜、眼睛、耳朵、鼻子内部、口腔内部、嘴唇、尿道开口、阴道、肛门、舌头、舌系带、毛发、牙齿、骨骼、泪腺、鼻窦粘膜、呼吸道、牙龈等。在一些实施例中,组织表面是皮肤表面或粘膜表面。在一些实施例中,粘膜表面可以是眼睛。在一些实施例中,粘膜表面可以是口腔、颊腔或阴道腔。
在一些实施例中,该方法进一步包括通过成像来检测抗体或抗体片段。抗体或其片段可以被本文公开的任何检测部分标记。在一些实施例中,成像包括通过SPECT(单光子发射计算机断层扫描)、PET(正电子发射断层扫描)、γ相机成像、直线扫描、放射自显影、磁共振成像、荧光成像、超声、及其组合来检测抗体或抗体片段。
在一些实施例中,本文公开的方法可用于非侵入性地测量体液的pH。在其他实施例中,本文公开的方法可用于针对功效监测组织药物水平。在其他实施例中,这些方法可用于检测凝血参数,例如凝血酶原时间(PT)、活化部分凝血活酶时间(aPTT)和凝血酶时间。在一些实施例中,这些方法可用于测试各种过敏。可以测试分离出的体液中的抗过敏原抗体,这些过敏原例如花粉、花粉、霉菌、宠物毛屑、尘螨和食物。
在一些实施例中,例如与可穿戴传感器结合,本文公开的方法可以提高代谢物和电解质的非侵入性监测的灵敏度和范围。
在一些实施例中,本文公开的方法可用于诊断塞扎里综合症(sezary syndrome)、蕈样霉菌病、皮肤损害、传染剂、肝炎、细菌感染、病毒感染、真菌感染、肺栓塞和肺结核。
在一些实施例中,这些方法可用于在切除后使黑色素瘤细胞成像,以监测微小浸润。在其他实施例中,方法和组合物可以与结肠镜检查结合使用。例如,可以在结肠清洗后使用包含细胞外基质组分的组合物,使用荧光标记的抗体对结肠癌进行成像。这样的方法可能导致标记抗体的渗透增加,从而在结肠癌抗原存在下导致信号增强。
在一些实施例中,由于非侵入性程序,本文公开的方法具有儿科应用。
这些实施例的方法可以包括各种附加步骤,包括例如在施加部位清洁表面组织等。例如,在施用透皮贴剂之前,通过电磁辐射、激光、皮肤磨损、化学剥离、超声、加热、冷却或通过针来烧蚀组织表面。例如,可以在施用透皮贴剂之前将皮肤准备擦拭物施用至皮肤。通常通过按摩、擦拭、填充、揉搓或任何其他方法将其应用于目标皮肤区域,以清洁目标皮肤部位并增加孔隙度。擦拭物还可含有本领域已知的常规皮肤渗透促进剂。擦拭物可以由浸泡在试剂中的纸、棉或基于纺织品的底物构成,这些试剂含有水、磷酸盐缓冲盐水、乳酸、肥皂、表面活性剂、或可在任何皮肤预处理程序(例如超声皮肤渗透系统(Sontra医药公司(Sontra Medical)))后用于清洁目标皮肤区域的任何其他化学物质、溶剂或其混合物。优选地,这些试剂是无机或有机溶剂,例如水、乙醇、异丙醇或其组合。该试剂的示例性配制品含有在水中的30%-95%异丙醇,并且该擦拭材料是纱布。
施用
在一些实施例中,活性剂和诱饵分子存在于同一组合物中。在一些实施例中,活性剂和诱饵分子存在于分开的组合物中。另外的实施例包括分别递送活性剂和诱饵分子的方法。在一些实施例中,包含活性剂的组合物和包含诱饵分子或透明质酸酶/弹性蛋白酶的组合物可以同时或顺序施用。一些实施例可包括将活性剂和诱饵分子共同施用于表面组织的步骤。例如,这样的方法可以包括将包含活性剂的组合物或配制品和包含诱饵分子的组合物或配制品施用于受试者的表面组织的步骤。在其他实施例中,可以在局部施用活性剂的组合物之前将诱饵分子的组合物施用于表面组织。例如,含有包含一种或多种诱饵分子的组合物的擦拭物可用于将诱饵分子施用到表面组织上,然后是将活性剂的组合物局部施用于表面组织的步骤。在其他实施例中,可将活性剂的组合物施用于表面组织,然后将诱饵分子的组合物施用于表面组织。
上述实施例的组合物可以增强已知局部活性剂的强度,从而减少达到治疗有效量所需的必要剂量。例如,在一些实施例中,含有活性剂和诱饵分子的组合物的强度可以比标准局部配制品中递送的活性剂高约等于80%或90%。在其他实施例中,含有活性剂和诱饵分子的组合物的强度可以比标准局部配制品中递送的活性剂高约等于约75%、约1.0%至约80%、约1.0%至约75%、约1.0%至约50%、约1.0%至约25%、约2.0%至约80%、约2.0%至约75%、约2.0%至约50%、约2.0%至约25%、约5.0%至约50%、约5.0%至约25%。因此,本文所述的组合物可提供已知局部施用的活性剂的治疗等效性,其施用剂量等于标准剂量或比标准剂量低至少约75%,等于标准剂量或比标准剂量低至少约50%,等于标准剂量或比标准剂量低至少约25%,等于标准剂量或比标准剂量低至少约10%,比标准剂量低约1.0%至约75%,比标准剂量低约1.0%至约50%,约比标准剂量低1.0%至约25%,比标准剂量低约1.0%至约10%,比标准剂量低约2.0%至约75%,比标准剂量低约2.0%至约50%,比标准剂量低约2.0%至约25%,比标准剂量低约2.0%至约10%,或这些实例范围所涵盖的任何范围或单个值。
在一些实施例中,与活性剂不和诱饵分子一起施用时相比,本文公开的方法当和诱饵分子一起施用时可递送多至少10%的活性剂、多至少15%的活性剂、多至少20%的活性剂、多至少30%的活性剂、多至少40%的活性剂、多至少50%的活性剂、多至少60%的活性剂、多至少70%的活性剂、多至少80%的活性剂、多至少100%的活性剂、多至少150%的活性剂、多至少200%的活性剂、多至少500%的活性剂、或多至少700%的活性剂。在一些实施例中,与活性剂不和诱饵分子一起施用时相比,当和诱饵分子一起施用时递送的活性剂的量可以多约10%至30%、多约10%至50%、多约10%至70%、多约10%至100%、多约10%至150%、多约10%至200%、多约10%至500%、或多约10%至1000%。
本文公开的组合物可以更有效地递送活性剂。即,在施用部位递送的活性剂的有效量比不存在诱饵分子下的活性剂的递送的有效量大得多。如实例1-15和17-20所示,诱饵分子出乎意料地有助于活性剂更有效地穿透组织,即使当活性剂以低浓度使用时。
在一些实施例中,组合物的施用是通过局部施用、透皮、经皮或微针注射进行的。施用还可以是例如静脉内、腹膜内、真皮下、皮下、真皮内、经皮、肌肉内、口服、关节内、肠胃外、鼻内或通过吸入。因此,合适的施用部位包括但不限于皮肤、支气管、胃肠道、眼睛、颊腔和耳朵。在一些实施例中,本文公开的组合物可经由针施用于任何实体组织。这样的组织包括肝组织、肺组织、胃肠道组织、肌肉组织、神经组织、骨骼、颊组织等。
可以经皮施用本文所述的化合物,即通过身体表面以及包括上皮和粘膜组织的身体通道内层施用。在一些实施例中,本文所述的组合物能以洗剂、霜剂、泡沫、贴剂、悬浮液、溶液和栓剂(直肠和阴道)的形式施用。在一些实施例中,透皮施用可以通过使用含有组合物以及对组合物呈惰性、对皮肤无毒并与组合物结合的载体的透皮贴剂来完成,以允许治疗性细胞穿透真皮和表皮以下。载体可以采用任意数量的以下形式,例如糊剂、霜剂、软膏、糊剂、凝胶和封闭装置。霜剂和软膏可以是水包油型或油包水型粘性液体或半固体乳剂。由分散于含有化合物的凡士林或亲水性凡士林中的吸收性粉末组成的糊剂也可能是合适的。可以使用各种封闭装置,例如覆盖含有组合物与或不与载体的储器的半透膜、或含有组合物的基质。其他封闭装置是文献中已知的。
本文公开的组合物还能以细雾喷雾剂的形式施用,该细雾喷雾剂包含纳米囊化的和非纳米囊化的治疗性细胞和细胞外基质组分并悬浮在载体中,以局部施用在人和动物的伤口和烧伤上。为此,将使用治疗性细胞,将其作为悬浮液掺入适当的载体中,以通过细雾雾化器(“GMSP预压缩细雾喷雾泵”类型)施用。
本文公开的方法可用于局部递送治疗性细胞,使得这些细胞穿透上皮层并均匀分布在下面的组织中。治疗性细胞的分布可根据组合物中细胞和细胞外基质组分的浓度而变化。在一些实施例中,治疗性细胞可以不均匀地分布,其中一些区域具有比其他区域更高的细胞浓度。在一些实施例中,可通过调节组合物中细胞外基质内容物的浓度来控制细胞穿过表皮层并进入下面的组织中。例如,当该组合物局部施用时,相对较低浓度的细胞外基质组分可以允许治疗性细胞部分地穿过表皮转运,而较高浓度的细胞外基质组分可以允许治疗性细胞完全穿过表皮转运至组织下层的基底膜(例如真皮和皮下组织)。
本文公开的方法可允许治疗性细胞穿透组织并重新繁殖并实现所需功能。例如,干细胞的局部施用可导致细胞穿透进入组织、分化、以及足以逆转潜在障碍的分化细胞的重新繁殖。在其他实施例中,所施用的抗原呈递细胞可以将免疫细胞募集到施用部位,并且可以帮助促进针对细菌或病毒感染或针对肿瘤细胞的有效免疫应答。
在一些实施例中,通过微针注射来施用该组合物。微针是空心针,其暴露高度在约0和1mm之间,总长度在约0.3mm至约2.5mm之间。优选地,微针是长度小于约2.5mm的空心针。最优选地,微针是长度小于约1.7mm的空心针。包含治疗性细胞和细胞外基质组分的组合物通过微针被递送到皮肤中至少约0.3mm且不超过约2.5mm的深度。
这些实施例的方法可以包括各种附加步骤,包括例如在施加部位清洁表面组织等。在这样的实施例中,组合物可以每天一次或多次施用于表面组织,并且施用可以进行至少1个月、2个月、3个月、4个月、6个月、8个月或12个月。
在这样的实施例中,组合物可以每天一次或多次施用于表面组织,并且施用可以进行至少1个月、2个月、3个月、4个月、6个月、8个月或12个月。在一些实施例中,组合物可根据需要施用一次、每日一次、每日两次、每日三次、每周一次、每周两次、每隔一周、每隔一天等。给药周期可包括施用约1周、约2周、约3周、约4周、约5周、约6周、约7周、约8周、约9周或约10周。在该周期之后,随后的周期可以在约1、2、3、4、5、6、7、8、9、10、11或12周后开始。治疗方案可包括1、2、3、4、5或6个周期,每个周期间隔开约1、2、3、4、5、6、7、8、9、10、11或12周。
本文公开的方法可以包括各种附加步骤,包括例如在施加部位清洁表面组织等。例如,在施用组合物之前,通过电磁辐射、激光、皮肤磨损、化学剥离、超声、加热、冷却或通过针来烧蚀组织表面。
皮肤外层或表皮的磨损(皮肤磨损)对于平滑或混合疤痕,瑕疵或可能由例如痤疮、日晒和老化引起的其他皮肤状况是合乎需要的。用于磨损皮肤的标准技术通常被分成两个领域,称为皮肤磨削和微晶皮肤磨削。两种技术都去除了称为角质层的表皮部分,身体将其解释为轻度损伤。然后身体取代损失的皮肤细胞,产生新的皮肤外层。此外,尽管存在与手术相关的轻度水肿和红斑,但由于新的皮肤外层,皮肤看起来和感觉更光滑。
微晶皮肤磨削通常是指由于通过发射沙子或沙砾流的手持件进行机械摩擦而去除皮肤表面的过程。例如,手持件可用于引导含有微小氧化铝、氯化钠或碳酸氢钠晶体的气流。随着手持件的每次通过,沙砾的动量趋向于磨损皮肤的两到三个细胞层。可替代地,新的“无晶体”微晶皮肤磨削技术使用没有沙砾流的金刚石尖头手持件。
在一些实施例中,在施用组合物之前,用电磁辐射烧蚀组织表面,例如使用所谓的点阵激光治疗。举例来说,这种方法采用电磁辐射(EMR),其具有在约1,850与100,000纳米之间的一个或多个波长并且具有在约1飞秒(1×10-15s)与10毫秒(10×10-3s)之间的脉冲宽度,注量范围为约1J/cm2至300J/cm2。在其他实例中,将组织用具有约2,200与5,000纳米之间的一个或多个波长的电磁辐射进行烧蚀。在仍其他实例中,将组织用具有约190与320纳米之间的一个或多个波长的电磁辐射进行烧蚀,注量范围为1J/cm2至300J/cm2。任选地,所选的用于烧蚀组织部分的条件使组织损伤的凝固区域最小化,例如通过将凝固区域保持在围绕烧蚀空隙的相对较小的直径。
电磁辐射(EMR),特别是以激光或其他光学辐射的形式,已经用于各种化妆品和医学应用,包括用于皮肤病学、牙科学、眼科学、妇科学、五官科学和内科学。对于大多数皮肤病学应用,可以使用将EMR递送至一个或多个靶组织表面的装置进行EMR治疗。EMR治疗通常被设计成(a)向组织递送一个或多个特定波长(或特定连续波长范围)的能量以诱导特定的化学反应,(b)向组织递送能量以引起温度升高或(c)向组织递送能量以损坏或破坏细胞或细胞外结构,例如用于皮肤重塑。在美容手术(例如皮肤再生)期间用于治疗皮肤的装置的实例包括LuxIR、1540、1440和2940点阵式手持件、ReliantSR激光仪以及科医人公司(Lumenis)、阿尔玛激光公司(Alma Lasers)、斯科顿公司(Sciton)的类似装置和许多其他装置。
在一些实施例中,本文公开的方法和组合物可用于与光动力疗法组合使用。光动力疗法是一种微创两步医疗手术,其使用称为光敏剂的可光活化药物来治疗一系列疾病。首先,施用光敏剂,并且一旦其渗透靶组织,然后通过暴露于特定波长的一定剂量的电磁(通常是光)辐射来激活光敏剂。本文披露的组合物可含有光敏剂。
在一些实施例中,本文可使用任何合适的光敏剂或试剂混合物。通常,这些将吸收约380nm至约900nm范围内的辐射。如本文所用,“光敏剂(photosensitizer/photosensitizing agent)”优选意指当与特定波长的辐射接触时形成单线态氧或热能的化合物。光敏剂的非限制性实例包括氨基乙酰丙酸酯、卟啉、卟啉衍生物、菌绿素、异菌绿素、酞菁、萘酞菁、焦脱镁叶绿酸、sapphyrin、得克萨卟啉、四氢卟吩、紫茜素、卟烯、吩噻嗪和金属络合物,例如但不限于锡、铝、锌、镥和锡本紫红素乙酯(SnET2)及其组合。
气雾剂递送
在一些实施例中,本文公开的组合物可以经鼻内递送。在一些实施例中,这些组合物可以是气雾剂配制品。在本文中,术语“气雾剂配制品”可以指水性组合物、干粉组合物或基于推进剂的组合物。气雾剂配制品能以不同方式例如经鼻或经口或通过吸入递送给受试者。
在一些实施例中,组合物可以是适于通过喷雾器(包括喷射喷雾器、振动网和静态网喷雾器、或孔喷雾器)进行肺递送的水溶液配制品。
在一些实施例中,组合物可以是包含微粉化颗粒的干粉,这些颗粒具有0.1至10微米的直径以及约0.5至4.5微米、约1至4微米、约1至3.5微米、约1.5至3.5微米、或约2至3微米之间的平均直径。干粉配制品适用于在干粉吸入器装置(DPI)或加压定量吸入器(pMDI)中使用。
在一些实施例中,本文公开的组合物可以呈基于推进剂的配制品的形式,其在本文中一般可以称为“pMDI配制品”。pMDI配制品适合通过例如加压定量吸入器(pMDI)等装置进行递送。在一些实施例中,组合物进一步包含推进剂。合适的推进剂是本领域已知的,并且包括例如卤素取代的烃,例如氟取代的甲烷、乙烷、丙烷、丁烷、环丙烷或环丁烷,特别是1,1,1,2-四氟乙烷(HFA134a)和1,1,1,2,3,3,3-七氟丙烷(HFA227)、或其混合物。
实例
尽管已经参考本发明的某些优选实施例相当详细地描述了本发明,但是其他版本也是可能的。因此,所附权利要求的精神和范围不应限于本说明书中包含的描述和优选变体。将参考以下非限制性实例说明本发明的各个方面。
实例1
透明质酸和仿生肽
制备含有促进毛发生长的肽混合物的组合物。以商品名出售的肽包括十肽-10、寡肽-54(CG-Nokkin)、十肽-18、乙酰基十肽-3和寡肽-42。通过将盐水中的肽与分子量为10,000道尔顿、20,000道尔顿、40,000道尔顿、60,000道尔顿或100,000道尔顿的透明质酸诱饵分子混合来制备肽组合物。对照配制品由单独的肽和单独的盐水组成。
图1A显示了使用具有完整角质层的皮肤进行研究的结果。这表明存在部分被动结合、受体介导的增强模式并且存在双峰特异性增强;随着尺寸的增加,非特异性水增强会增加,因此增强的渗透效果是特定的。即使存在死皮,添加逐渐增大的分子量也会逆转益处。
图1B显示了使用胶带剥离方法剥离角质层的皮肤进行研究的结果。这证明了跨没有角质层的可存活皮肤层的主动结合、受体介导的增强模式(即根本没有水增强作用)和基于MW存在的特异性增强;较大的MW不仅消除了增强作用,而且延缓了跨活皮肤层(代表深层表皮和真皮渗透的屏障)中细胞的渗透。
对于每种测试组合物,显示了相对于来自单独肽组合物的肽通量的肽通量百分比。每种组合物进行两次测试,第一次研究用实线表示,第二次研究用虚线表示。已知分子量高达300,000Da的透明质酸能够穿透皮肤。图1A-1B中的数据显示使用小于约40,000Da的透明质酸分子进行的肽递送是通过不同于大于40,000Da的透明质酸分子的递送途径,并且这两种递送途径与水合作用完全无关。当角质层存在于皮肤上时(图1A),在具有20,000Da和60,000Da的透明质酸的组合物情况下观察到肽递送的峰。当从皮肤上剥离角质层时(图1B),未观察到使用60,000Da透明质酸诱饵分子实现的峰,证明肽递送不仅仅归因于水合作用,因为皮肤水合作用引起的皮肤渗透增强会随着诱饵分子量的增加而增加。此外,由于已知100,000Da透明质酸可穿透角质层(Essendoubi,2016),如果从本发明组合物观察到的递送是由于水合作用,则可以预期观察到肽从具有100,000Da透明质酸诱饵分子的组合物的递送跨过具有和不具有角质层的皮肤。图1B显示具有100,000Da透明质酸诱饵分子的组合物提供的肽的递送少于具有分子量透明质酸的组合物。相对于没有诱饵分子的组合物的递送,具有40,000Da的诱饵分子的组合物具有不太增强的肽递送(图1A)。
实例2
透明质酸和水杨酸盐
制备含有1%水杨酸盐和1%透明质酸诱饵分子的组合物,该透明质酸诱饵分子具有四个分子量:小(5,000Da至10,000Da)、小至中(10,000Da至20,000Da)、低至中(20,000Da至30,000Da)以及中(30,000Da至40,000Da)。还制备了仅含有水杨酸盐的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池的接收侧水杨酸盐的浓度,结果示于图2中。
与来自仅有水杨酸盐的组合物中的水杨酸盐的通量相比,具有10,000Da至20,000Da的透明质酸诱饵分子组合物使水杨酸盐的通量高27%。与来自仅有水杨酸盐的组合物中的水杨酸盐的通量相比,20,000Da至30,000Da的诱饵分子使水杨酸盐皮肤通量增加约5%。
实例3
透明质酸和类固醇
制备含有1%氢化可的松和1%透明质酸诱饵分子的组合物,该透明质酸诱饵分子具有四个分子量:小(5,000Da至10,000Da)、小至中(10,000Da至20,000Da)、低至中(20,000Da至30,000Da)以及中(30,000Da至40,000Da)。还制备了仅含有氢化可的松的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池的接收侧水杨酸盐的浓度,结果示于图3中。
具有透明质酸诱饵分子的组合物增加了氢化可的松跨皮肤的递送,与来自缺乏诱饵分子的组合物的氢化可的松的通量相比,20,000Da至30,000Da的中等大小的诱饵分子将氢化可的松通量增加325%。与来自没有诱饵分子的组合物的氢化可的松的通量相比,分子量为约10,000Da至20,000Da的小到中尺寸诱饵分子使水杨酸盐皮肤通量增加约250%。
实例4
弹性蛋白和利多卡因
使用含有弹性蛋白诱饵分子的组合物评估利多卡因跨皮肤的递送。制备含有1wt.%利多卡因和在盐水中的0.5wt%弹性蛋白诱饵的组合物,该弹性蛋白诱饵具有三个分子量:非常非常小(2,000Da至5,000Da)、非常小(5,000Da至10,000Da)以及小(10,000Da至20,000Da)。
获得可存活的猪皮肤并用于产生中真皮移植物(0.045-0.055个单位)。将移植物定位在经皮通量装置中。将装置中的流量维持在最低设置,并且收集每次重复(对于每种测试配制品和对照配制品,n=8)的所有接收液。在样本应用并停留在供体皮肤表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化。使用光谱法测定澄清的匀浆溶液和流过样品的利多卡因含量。在12-20小时的渗透期后,测定皮肤中利多卡因的浓度。结果在如图4显示为应用的利多卡因的百分比。
没有诱饵分子的利多卡因配制品实现了3%渗透。添加具有小分子量(10,000Da至20,000Da)的弹性蛋白诱饵分子使皮肤渗透增加约7倍(渗透显著改善,p=0.0001)。
实例5
透明质酸和米诺环素
口服米诺环素HCl非常有效,但受到耳毒性和新出现的耐药性的限制。相对于口服,大多数医生会局部使用米诺环素。然而,因为米诺环素不能有效地穿过皮肤,局部应用目前不如口服有效。因此,必须使用更高的浓度,这些浓度会使皮肤和纺织品变色。
体外测量米诺环素向猪皮肤中的递送,并与米诺环素从米诺环素在盐水中的组合物(即没有诱饵分子)中的递送进行比较。制备含有1wt.%米诺环素和1%透明质酸诱饵分子的组合物,该透明质酸诱饵分子具有三个分子量:平均10,000Da、平均20,000Da和平均30,000Da。还制备了在盐水中含有1wt%米诺环素的对照配制品。
图5显示了研究结果,其中从米诺环素和透明质酸钠的局部配制品递送到猪皮肤移植物中的组织中米诺环素的量(μg米诺环素/g组织)用虚线填充柱显示,而从不含透明质酸的局部配制品递送到猪皮肤移植物中的组织中米诺环素的量用无填充柱显示。尽管米诺环素具有低的天然渗透性,但基于多糖的诱饵显著增强了渗透性(p=0.0004)。这些结果证实,诱饵介导的策略可以提供局部米诺环素的高渗透性。具有低分子量的诱饵分子使米诺环素非常低的基础渗透率增加到可以达到比口服更高的组织浓度的水平,同时避免变色和全身副作用。含有米诺环素和诱饵分子的局部组合物可用于治疗或改善皮肤结构感染或失调,例如蜂窝组织炎。
实例6
用于保护皮肤免受UVA/UVB射线的组合物
由于基质厚、与化妆品不相容、持续时间短,目前用于防晒的化学试剂的顺应性较差。通过增强现有试剂的功能,可以开发出更有效的防晒霜、防止擦掉的防晒霜、和/或与其他产品一起感觉和使用更理想的配制品(以诱导更好的顺应性)。
在该研究中,制备并测试了用于保护皮肤免受UV-A和/或UV-B暴露的组合物。在供体池中,组包括A)以1:10掺入盐水的理肤泉(Laroche Posay)Anthelios 60防晒剂(n=10个重复)、或B)以1:10掺入分子量为10,000的1%透明质酸钠的理肤泉Anthelios 60防晒剂(增强的Anthelios 60)(n=10个重复)。将流量维持在最低设置,并且收集每次重复的所有接收液。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后进行穿孔活检,置于96孔板中并在全范围UV光谱中使用。每组的UV吸光度由每组的波长确定,UVA和UVB值由适当的波长确定。结果显示在图6、图7A-7B和图8中。
添加本身没有UV吸光度的增强剂,在UVA(P=0.001)和UVB(P=0.001)两者中统计性地显著提高了商业上可获得的UV阻断剂混合物的性能,如图6所示。各组的各个波长结果显示在图8中,每组的一个代表性光谱显示在图7A和图7B中。
测试具有和不具有诱饵分子的组合物以确定皮肤中的UV吸收。图6是显示皮肤中UVA和UVB吸收的柱状图(4.0对应于100%),其中虚线填充柱对应于具有诱饵分子的防晒组合物,并且实心白条为仅防晒剂。
图7A-7B是市售防晒剂(Anthelios 60)(图7A)和具有诱饵分子的市售防晒剂(Anthelios 60)(增强的Anthelios 60)(图7B)的UV吸收随波长(以nm计)变化的图。
图8是显示市售防晒剂(Anthelios 60)(实线)和具有诱饵分子的市售防晒剂(Anthelios 60)(增强的Anthelios 60)(虚线)的通过皮肤的UV吸收百分比随波长(以nm计)变化的图。
实例7
透明质酸和加巴喷丁
使用猪皮肤移植物,体外测量在透明质酸情况下加巴喷丁向皮肤中的递送,并与加巴喷丁从加巴喷丁在盐水中的组合物(没有诱饵分子)中的递送进行比较。在供体池中,组由以下项组成:A)盐水中1%加巴喷丁(n=8个重复)、B)在盐水中1%加巴喷丁加1%透明质酸钠诱饵3,000Da(n=8个重复)、或C)仅盐水(n=8个重复)。
处理可存活的猪皮肤以产生中真皮移植物(0.045-0.055个单位),并将移植物定位在经皮通量装置中。将装置中的流量维持在最低设置,并且收集每次重复(对于每种测试配制品和对照配制品,n=8)的所有接收液。在样本应用并停留在供体皮肤表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后用于使用UPLC-质谱仪方法测定皮肤样品中的加巴喷丁含量。简而言之,将组织在55℃下在0.5mL的50%乙腈的去离子水中在搅拌下温育过夜。在分析之前,将校准标准品和组织提取溶剂样品在去离子水中稀释10倍。以1μL注射体积分析稀释的标准品和样品。浓度报告为μg加巴喷丁/g组织。
图9显示了研究结果,其中显示了从加巴喷丁和透明质酸钠的局部配制品和从不具有透明质酸钠的加巴喷丁配制品递送至猪皮肤移植物中的加巴喷丁的量(μg加巴喷丁/g组织)。单独的加巴喷丁不会产生高于盐水的显著渗透(p=0.99),但在诱饵存在下加巴喷丁相对于盐水(p=0.018)和单独的加巴喷丁(p=0.013)实现了显著的渗透。具体地,加巴喷丁单独产生0.09μg加巴喷丁/g组织的组织水平,而添加诱饵分子的加巴喷丁产生174.01μg加巴喷丁/g组织的组织水平。因此,添加诱饵分子使药剂向皮肤的递送增加了1,900倍,并且使局部加巴喷丁渗透统计学上显著增加。
实例8
透明质酸和棕榈酰-赖氨酸-苏氨酸-苏氨酸-赖氨酸-丝氨酸
制备含有化妆品试剂、作为诱饵分子的棕榈酰-赖氨酸-苏氨酸-苏氨酸-赖氨酸-丝氨酸(pal-KTTKS)和透明质酸钠(3,000Da)的局部组合物。组包括A)掺入Olay ProX中的1%Pal-KTTKS(n=8个重复),或B)掺入Olay ProX中的1%Pal-KTTKS加在盐水中的1%透明质酸钠诱饵3,000Da(n=8个重复)。
处理可存活的猪皮肤以产生中真皮移植物(0.045-0.055个单位),并将移植物定位在经皮通量装置中。将装置中的流量维持在最低设置,并且收集每次重复的所有接收液。在样本应用并停留在供体皮肤表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化。然后将澄清的匀浆溶液和流过样品用于使用UPLC-质谱仪方法测定皮肤样品中pal-KTTKS含量。
图10显示了研究结果,其中显示了从pal-KTTKS和透明质酸钠的局部配制品和从不具有透明质酸钠诱饵的pal-KTTKS局部配制品递送的组织中的pal-KTTKS的量(μg pal-KTTKS/50mg组织)。在12-20小时的渗透期后,单独pal-KTTKS的配制品(没有诱饵分子)产生约100μg的pal-KTTKS/50mg组织。诱饵分子的添加改善了试剂向皮肤中的渗透,接近450μgpal-KTTKS/50mg组织。因此,在试剂向皮肤的递送中在没有优化的情况下,向局部组合物中添加诱饵分子产生了通量增加近422%(P<0.01)。因此,在没有任何额外的配制品变化的情况下,多糖诱饵提供了最广泛认可的肽基护肤活性物质的渗透的实质性和显著增强。
实例9
含有诱饵的组合物中FITC-右旋糖酐的眼部递送
在全眼轮廓未受伤的情况下,获得完整的、新鲜的、可存活的猪眼。将眼睛在治疗溶液中沐浴至中线(镜片向下)过夜,同时通过视神经结扎悬挂在上方。组合物如下制备:A)盐水中的5,000Da FITC-右旋糖酐(n=2个重复)、B)盐水中3,000Da的1%透明质酸钠中的5,000Da FITC-右旋糖酐(n=2个重复)、C)盐水中的0.5%短弹性蛋白中5,000Da FTIC-右旋糖酐(n=2个重复)和D)仅盐水。
将眼睛在盐水中彻底洗涤5次,然后快速冷冻并用反射共聚焦成像系统(Vivascope 1500)分析,以非侵入性成像并可视化FITC-右旋糖酐的渗透。共聚焦显微镜显示,尽管镜片内几乎没有显著的信号,但多糖和肽基诱饵分子均可使FITC-右旋糖酐标记物(药物模型)可见地渗透到房水(包括前房和后房以及睫状体)、结构要素(包括小带和巩膜)以及玻璃体液(包括视网膜洗涤)中。盐水对照显示没有颗粒荧光,也没有药物(标志物)渗透,因为不存在FITC-右旋糖酐。
该实验证实,当与两类诱饵结合时,5,000Da药物标志物渗透到眼睛中。使用150,000MW的右旋糖酐和抗体标志物的类似实验证实了在两类诱饵情况下的渗透;尽管150,000MW相比于5,000MW的通量量级不同,但当局部应用于完整的眼睛时,两者都表现出渗透性。
实例10
从含有诱饵的组合物到指甲的FITC-右旋糖酐递送
将1%5,000Da FITC-右旋糖酐和1%10,000Da透明质酸钠的混合物以1:10稀释度加入市售的指甲基质中。将该材料施加到脚趾甲上并静置3小时。如前所述使用共聚焦成像来观察FITC-右旋糖酐向指甲板中的渗透。
以7微米的步长获得图像。
如预期的那样,指甲表面上存在非常高水平的信号。如通过粒状荧光图案显现的,观察到高水平的5,000Da FITC-右旋糖酐缀合物渗透到指甲板的最深层。因此,针对指甲所关注的大多数抗真菌和营养成分可以通过添加小诱饵片段来递送。
实例11
含有诱饵分子的组合物中水杨酸盐的粘膜递送
预期了用于将试剂递送至粘膜组织的组合物,并且使用可存活的猪颊组织进行研究以评估水杨酸盐从具有弹性蛋白诱饵分子的组合物进行的粘膜渗透。准备以下组合物用于测试:A)盐水中1%水杨酸钠(n=4个重复)、或B)盐水中的1%水杨酸钠加0.5%短弹性蛋白片段诱饵(诱饵)(n=4个重复)。
获得可存活的猪颊组织并产生移植物。将移植物置于经皮通量装置中以测量粘膜穿透。将装置中的流量维持在最低设置,并且收集每次重复(对于每种测试配制品和对照配制品,n=8)的所有接收液。在样本应用并停留在供体粘膜组织的情况下,继续通量12-20小时。在12-20小时的测试期后,洗涤来自每个池的组织,然后均质化。然后将澄清的匀浆溶液和流过样品用于通过吸光度测定水杨酸盐含量。来自具有弹性蛋白诱饵的组合物和来自没有诱饵的组合物的水杨酸盐的皮肤渗透性显示在图11中。
这些结果表明,向组合物中添加诱饵分子使水杨酸盐的粘膜渗透增加350%。
实例12
含有诱饵分子的组合物中抗体的递送
制备由以下组成的组合物:在供体池中,A)盐水中的25μl碱性磷酸酶缀合的IgG抗体(n=8个重复)、B)盐水中的25μl碱性磷酸酶缀合的IgG抗体加3,000Da的1%透明质酸钠(n=8个重复)、C)盐水中的25μl碱性磷酸酶缀合的IgG抗体加5,000Da的1%透明质酸钠(n=8个重复)、或D)盐水中的25μl碱性磷酸酶缀合的IgG抗体加10,000Da的1%透明质酸钠(n=8)。
处理可存活的猪皮肤以产生中真皮移植物(0.045-0.055个单位),并将移植物定位在经皮通量装置中。将流量维持在最低设置,并且收集每次重复的所有接收液。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后将流过样品用于通过吸光度测定碱性磷酸酶含量。结果如图12所示。
如流经液中的碱性磷酸酶活性所测量,单独的抗体没有表现出显著的渗透,而诱饵介导的渗透实现了施加负载量的超过2%的渗透。因此,仅通过添加诱饵分子就可以实现统计学上显著的渗透增加(P=0.003)。因此,该方法提供高百分比渗透,这使得能够开发局部大分子治疗剂。鉴于该抗体标记为150-160KD,实际上任何衍生化的抗体或抗体片段的递送是可行的,类似分子(如,肉毒杆菌毒素及其衍生物或嵌合体)的递送同样也是可行的。
实例13
功能性抗氧化能力
透明质酸(HA)和弹性蛋白(E6)诱饵都提供了来自几种不同配制品的专有抗氧化剂混合物的渗透性增加。将相同的抗氧化剂混合物与如下详述的几种不同媒介物和诱饵组合施用于皮肤。导致对过量功能性氧化应激的抗性增加。
用植皮刀处理扩散室可存活的猪皮肤至中真皮厚度,然后以n=6/预期条件进行穿孔活检。准备改进的6区扩散池装置并设定流速为0.022ml/min。将配制品(各200μl)施用于顶部(供体)表面并按摩。对于这些实验,对于每个池收集接收液12小时,然后去除皮肤,清洁并快速冷冻以用于将来在盐水中冷均质化。
应用于猪皮肤的配制品
·盐水
·配制品1
·配制品1+1%HA
·配制品1+0.5%E6(VGVAPG)
·配制品2
·配制品2+1%HA
·配制品2+0.5%E6
·配制品3
·配制品3+1%HA
·配制品3+0.5%E6
·配制品3+1%HA+0.5%E6
Invitrogen Amplex红色试剂盒(目录号A22188):在HRP存在下,Red试剂(10-乙酰基-3,7-二羟基吩噁嗪)与H2O2以1:1的化学计量比反应,产生红色荧光氧化产物试卤灵。我们使用该试剂盒作为活性氧物质的基线测量(如设计试剂盒)以确保不存在异常的ROS基线值。然后我们故意引入氧化应激并观察每个流过样品如何响应。按照试剂盒说明进行溶液制备和反应设置。
将反应在30℃温育30分钟,避光并每5分钟混合5秒(在读板器中)。测量260nm处(参考值以确保正态性)和560nm处(试卤灵)的吸光度并记录值作为基线(预应力)。选择吸光度而不是荧光,以便在掺入后(post-spike)更快地读取(每个循环约1分钟)。对于每个点,减去从零H2O2对照孔的平均值得到的值(n=2)。
向每个孔中快速掺入20uL的0.1mM H2O2原液,然后测量260nm和560nm处的吸光度,并记录值作为应激时间零。连续测量动态循环,共5个循环(约5分钟),然后在10min和15min时再次测量。由于来自掺入的过量H2O2,试卤灵本身可以经历第二次氧化成非吸收/荧光状态,多次读取是为了确保可以评估峰值和线性范围。
与正常皮肤对照(盐水处理的)相比,配制品1的配制品达到平均5.15%的抗氧化能力。虽然没有统计学显著性意义(p>0.2),但配制品1处理的皮肤的抗氧化能力始终大于盐水处理的皮肤。
所有随后的配制品比较均相对于配制品1配制品作为参考抗氧化剂能力进行。通过这种方式,可以在不直接测量单个物质的情况下评估能力相比于当前基质的增加。
HA配制品:HA增加了针对每种基质的接收液的抗氧化能力,但是从配制品到配制品之间存在显著差异:
总体而言,当将HA加入配制品3基质中时,观察到最高的显著抗氧化能力增加。
与配制品1皮肤相比,E6提供了抗氧化能力的一致增加,但由于样本量小和增加较少,因此没有达到p<0.05(大多数p<0.08)。由于sans基质是围绕HA行为而不是E6设计的,因此从配制品到配制品的E6增强没有显著差异。与先前在其他配制品基质中观察到的其他活性物质不同,E6没有达到与HA观察到的抗氧化能力一样的高增加。
实例14
FTIC-右旋糖酐共聚焦皮肤分析
使用1500对人受试者进行实时共聚焦显微镜成像,以显示高达150,000Da的各种尺寸的FITC-右旋糖酐缀合物跨毛发皮肤(前臂背侧)和非毛发皮肤(前臂掌侧)的渗透。在盐水中制备各组,各组由以下组成:盐水中的1%5,000Da的FITC-右旋糖酐或1%5,000Da FITC-右旋糖酐加1%透明质酸钠,该透明质酸钠平均分子量为5,000Da至20,000Da。使用分子量为10,000Da至20,000Da的0.5%弹性蛋白片段(E6)代替HA诱饵获得了类似的结果。
实例15
总结
应理解,实施例1-6、11、12说明透明质酸作为本文考虑的诱饵分子的诱饵分子示例。如上所述,考虑了胶原和弹性蛋白诱饵分子,其中可以选择诱饵分子的分子量以调整相关试剂跨皮肤的递送。下表总结了诱饵分子(使用透明质酸作为示例)对小分子化合物(例如,分子量小于约850Da)的透皮递送、对大分子化合物(例如,肽或蛋白质)的透皮递送、对局部施用时诱饵分子渗透到皮肤中的程度、以及对诱饵分子增加皮肤含水量的影响,使用+符号来反映效果的程度。如图所示,诱饵分子的皮肤渗透、诱饵分子引起的皮肤水合以及化合物向皮肤中的递送之间存在脱节,表明皮肤递送的增强不是由于水合作用或诱饵的存在,而是由于皮肤中的诱饵分子的活性。
虽然上面已经讨论了许多示例性方面和实施例,但是本领域技术人员将意识到其某些修改、置换、添加和子组合。因此,以下所附权利要求和此后引入的权利要求旨在被解释为包括在其真实精神和范围内的所有这些修改、置换、添加和子组合。
实例16
与不含诱饵分子的组合物相比,将制备以下化合物并测试其增加的通量:
胶原和维生素C
将制备含有维生素C和胶原诱饵分子的组合物,该胶原诱饵分子具有在盐水中标记为A1、B1、C1的三个分子量。还将测试由盐水中的维生素C组成的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池接收侧的维生素C浓度。
胶原和双氯芬酸
将制备含有双氯芬酸和胶原蛋白诱饵分子的组合物,该胶原蛋白诱饵分子在盐水中具有三个分子量5,000Da、25,000Da和50,000Da。还将测试由盐水中的双氯芬酸组成的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池接收侧的双氯芬酸浓度。
弹性蛋白和烟酰胺
将制备含有烟酰胺和弹性蛋白诱饵分子的组合物,该弹性蛋白诱饵分子在盐水中具有三个分子量5,000Da、25,000Da和50,000Da。还将测试由盐水中的烟酰胺组成的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池接收侧的烟酰胺浓度。
弹性蛋白和萘普生
将制备含有萘普生和弹性蛋白诱饵分子的组合物,该弹性蛋白诱饵分子在盐水中具有三个分子量5,000Da、25,000Da和50,000Da。还将测试含有在盐水中的萘普生的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池接收侧的萘普生浓度。
用于毛发生长的比马前列素的局部施用
将制备包含0.01%比马前列素和0.5%弹性蛋白片段诱饵分子的组合物,该弹性蛋白片段诱饵分子的分子量范围为:小(5,000Da至10,000Da)、小至中(10,000Da至20,000Da)、低至中(20,000Da至30,000Da)以及中(30,000Da至40,000Da)。还将制备仅含有0.01%比马前列素和仅含盐水的对照配制品。该组合物将应用于最近在大约21天前完成化疗周期并且经历接近全头皮脱发的受试者。与可比对照相比,预期用含有诱饵的组合物治疗的受试者在1、2和4周时实现更快的毛发生长速率。另外,在用含有诱饵的组合物治疗的受试者中,预期头发的长度、厚度和密度更大。
针对毛干的诱饵增强色彩处理
制备含有市售染发剂配制品的组合物的掺入1%的具有低至中分子量(20,000Da至30,000Da)的透明质酸诱饵分子的组合物,并与单独的染料比较。将组合物各自施用于一半头皮毛干(受试者内对照),并在30分钟后移除。冲洗后,一周后和4周后评估颜色深度。用含有诱饵的组合物处理的毛干预期显示出更大的色彩丰富度、深度和持久性。
用于干眼症的比马前列素
将制备包含0.01%比马前列素和0.5%弹性蛋白片段诱饵分子的组合物,该弹性蛋白片段诱饵分子的分子量范围为:小(5,000Da至10,000Da)、小至中(10,000Da至20,000Da)、低至中(20,000Da至30,000Da)以及中(30,000Da至40,000Da)。还将制备仅含有0.01%比马前列素和仅含盐水的对照配制品。这些组合物将以滴眼剂的形式施用于受试者的眼睛表面。相对于可比较的对照,预期用含有诱饵的组合物治疗的受试者会从干眼症中恢复。
实例17
局部肽基诱饵介导的水杨酸盐通过粘膜表面递送的增强
收获猪颊组织并清除其他组织。根据相关方案获得穿孔活检(每个n=4),并在透皮通量装置中使用。各组由以下组成:(1)生理盐水中的0.5%弹性蛋白肽E6+1%水杨酸盐;(2)生理盐水中1%的水杨酸盐;(3)仅盐水。在样本应用并停留在供体表面的情况下,继续通量12-20小时。通过酶标仪(美谷分子仪器公司(Molecular Devices)Spectramax M3)分析流经样品的水杨酸盐含量。肽基诱饵E6的添加使水杨酸盐从颊组织到流经液的渗透增加了351.8%。尽管单独的水杨酸盐显示穿透组织,但总的渗透率与单独的盐水相比在统计学上不显著。然而,将诱饵添加到水杨酸盐中具有统计学上的显著增加(p=0.036)。
实例18
西地那非的局部应用
根据相关方案,在透皮通量装置中采用中真皮移植物的复制品。各组由以下组成:(1)生理盐水中的1%寡糖和多糖诱饵(长度很短的透明质酸链)+1%西地那非(n=10);(2生理盐水中1%西地那非(n=10);和(3)仅盐水(n=2)。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化;测定澄清的匀浆溶液中西地那非的浓度,并通过美谷分子仪器公司Spectramax M3在96孔微孔板测定中读取。与单独的生理盐水相比,单独的西地那非不能被检测到并且没有显著性;西地那非+HA以13.8%的通量渗透,以达到组织浓度为96.6ug/移植物。对于具有诱饵的西地那非相比单独西地那非的更高的渗透率,结果是高度统计学上显著的(p=0.0003)。考虑到口服西地那非的生物利用度为41%,并假设组织均匀分布,目前批准的西地那非的治疗剂量在平均75kg的男性中仅能达到0.137ug西地那非/g组织的组织浓度。因此,添加诱饵将提供一种局部用药,其可以潜在地减少总暴露量和全身暴露量(风险和副作用),并获得比口服施用甚至更高的功效。
实例19
糖基诱饵局部增强毛干渗透
获得单个供体毛干,用盐水洗涤,并分成7cm的段。每个段(每组n=5)在各自的处理溶液中温育5分钟;各组由以下组成:(1)生理盐水中的1.0%寡糖和多糖诱饵(长度很短的透明质酸链)+1%FITC-右旋糖酐(MW 3,000-5,000);(2)生理盐水中的1%FITC-右旋糖酐(MW 3,000-5,000);和(3)仅盐水。将毛干用过量的盐水洗涤5次,然后在96孔板上分成1cm的段(每孔七个段)。然后通过酶标仪(美谷分子仪器公司Spectramax M3)测量FITC的吸光度。尽管单独的右旋糖酐明显粘附在毛干的外层上以达到与盐水相比在495至518nm处的吸光度增加,但是由于其到毛干的渗透增加,添加短寡糖和多糖诱饵使在5分钟的短暂治疗期内达到234%的负载率。显微镜确认结果。
实例20
通过糖基/肽基诱饵局部增强骨和骨膜渗透。
获得猪骨头并仔细清洁骨膜外的所有软组织。每个骨头都具有一部分反射的骨膜以直接暴露骨,并且相邻区域保留了完整的骨膜,以评估骨渗透性和骨膜的渗透性。要测试的区域的边缘区域应用了凡士林来创建储器。将2ml的测试品施加到包括完整骨膜和暴露的骨表面的骨头上的每个测试区域。各组由以下组成:(1)生理盐水中的1.0%寡糖和多糖诱饵(长度很短的透明质酸链)+1%FITC-右旋糖酐(MW 3,000-5,000)组成;(2)仅生理盐水中的1%FITC-右旋糖酐(MW 3,000-5,000);和(3)生理盐水中的1.0%弹性蛋白片段诱饵+1%FITC-右旋糖酐(MW 3,000-5,000)。使骨头温育过夜,然后用大量过量的盐水洗涤五次,然后进行UV曝光、照相和图像分析。尽管具有诱饵的组显示出高的多且更均匀的水平,但所有组均显示出FITC-右旋糖酐与骨膜的结合和保留。相反,具有诱饵的两组均显示出FITC-右旋糖酐到骨皮质中的高渗透和保留,而没有诱饵的组则基本上不可检测。
实例21
较大分子量的诱饵消除了通量
根据相关方案,将中真皮移植物的复制品(根据Illustris标准作业程序,针对通量装置的移植物和打孔器)用于透皮通量装置。各组由以下组成:(1)生理盐水中1%寡糖和多糖诱饵-20K MW以下的透明质酸链+1%水杨酸钠(n=6);(2)生理盐水中0.9%寡糖和多糖诱饵-40K Mw以下的透明质酸链+0.1%的150K Mw以上的透明质酸链+1%水杨酸钠(n=6);(3)+生理盐水中1%水杨酸钠(n=6);和(4)仅生理盐水(n=6)。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化;测定澄清的匀浆溶液中水杨酸钠的浓度,并通过美谷分子仪器公司Spectramax M3在96孔微孔板测定中读取。水杨酸钠的OD290比盐水高0.0165AU,因此仅表现出低水平的可检测渗透。单独的低MW水杨酸钠作为诱饵(无大诱饵)的OD值为0.1075AU(增加651.5%)。有趣的是,在保持总HA不变的情况下,将一小部分高MW HA添加到低MW HA中可消除诱饵的所有优势;具有少量高MW HA的诱饵并没有比单独的水杨酸钠显著好得多,并且实际上在数值上更接近盐水(降低了-0.01AU,但不是统计学上显著的)。因此,添加少量高MW HA完全消除了低MW HA诱饵的益处。该结果解释了先前的观察结果,即低纯度小MW HA(被高MW HA污染)并没有表明纯的小MW HA所观察到的益处。
实例22
较大的HA消除了通量–向纯的低MW成分中添加高MW“杂质”
根据相关方案,将中真皮移植物的复制品(根据Illustris标准作业程序,针对通量装置的移植物和打孔器)用于透皮通量装置。各组由以下组成:(1)生理盐水中的1%寡糖和多糖诱饵-20K MW以下的透明质酸链(经UPLC确认不含可检测的MW>50K)+1%FITC-右旋糖酐(MW 150K)(n=6);(2)1%寡糖和多糖诱饵-20K MW以下的透明质酸链+1%FITC-右旋糖酐(MW 150K)+“作为污染物添加”的0.1%的60K MW的透明质酸链(n=6);(3)1%寡糖和多糖诱饵-20K MW以下的透明质酸链+1%FITC-右旋糖酐(MW 150K)+“作为污染物添加”的0.1%的100K MW的透明质酸链(n=6);和(4)生理盐水中的1%FITC-右旋糖酐(MW 150K)(n=6)。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化;通过荧光测定澄清的匀浆溶液的FITC浓度,如通过美谷分子仪器公司Spectramax M3在96孔微板中读取的。结果呈现在下表中。
与现有文献一致,盐水中的MW 150KD分子处于背景水平,因此盐水中的1%FITC-右旋糖酐(MW 150K)没有给出可检测的通量。相反,单独的1%的纯的低MW FITC-右旋糖酐(MW 150K)作为诱饵(无大诱饵)提供了可检测的、统计上显著的通量,超过18%。有趣的是,将相对一小部分高MW HA添加到低MW HA中消除了诱饵的所有显著优势;具有少量高MW HA的诱饵并不比单独的盐水或背景显著好得多,并且在数值上接近背景水平。因此,将少量高MW HA添加至其他功能性诱饵混合物中在统计学上完全消除了低MW HA诱饵的益处。该结果解释了先前的观察结果,即低纯度小MW HA(被高MW HA污染)并没有表明纯的小MW HA所观察到的益处。
实例23
较大的HA消除了通量–从具有低平均MW的宽MW群体中去除高MW“杂质”
为了复制现实世界的制造条件(xxx=我们应该对这些条件作一些要求)并进一步阐明实例1中的观察结果,通过降解非常大的MW HA(MW>1,800,000;来自Habier公司),产生了低平均MW HA,使得又宽又平的MW范围实现平均不到20K。具体地,将18mg(约300个单位/mg)透明质酸酶(来自西格玛公司(Sigma))添加至1L的3%透明质酸溶液中,并在37摄氏度下轻轻摇动72小时。将产物加热灭活以破坏残留的透明质酸酶,进行乙醇沉淀并真空干燥。将产物通过UPLC进行测试,发现宽范围的平均18K的MW。原样制备这种低平均HA MW混合物的样品;首先通过50K的MW截止过滤器(Amicon过滤器)处理其他样品,使得其他样品只有50K及以下的MW。过滤不会改变计算的平均MW,但会截断曲线,使得低MW HA不再包含任何可检测的高MW杂质。
根据相关方案,将中真皮移植物的复制品(根据Illustris标准作业程序,针对通量装置的移植物和打孔器)用于透皮通量装置。各组由以下组成:(1)生理盐水中的纯净混合的1%寡糖和多糖诱饵-18K MW以下的透明质酸链(经UPLC确认不含可检测的MW>50K)+1%FITC-右旋糖酐(MW150K)(n=6);(2)未纯化的1%寡糖和多糖诱饵-20K MW以下的透明质酸链(其包含MW>50K的工艺污染物/杂质)+1%FITC-右旋糖酐(MW 150K)+“作为污染物添加”的0.1%的60K MW的透明质酸链(n=6);和(3)生理盐水中的1%FITC-右旋糖酐(MW150K)(n=6)。在样本应用并停留在供体表面的情况下,继续通量12-20小时。洗涤每个池(每个室)的皮肤,然后均质化;通过荧光测定澄清的匀浆溶液的FITC浓度,如通过美谷分子仪器公司Spectramax M3在96孔微板中读取的。结果呈现在下表中。
实例24
概念验证150,000Da面部应用
方法.使用3000对人受试者进行实时共聚焦显微镜成像,以可视化150,000Da FITC-右旋糖酐缀合物(模拟大分子量分子,如肉毒杆菌毒素A)在眉间和外侧眼角区域的渗透。将100μL等分试样的生理盐水中的1%FITC-右旋糖酐(平均分子量为150,000Da)+1%多糖诱饵施加到指定区域并轻轻按摩。避免阳光直射,并放置1小时。按照常规的共聚焦方法,通过用医用酒精片轻轻清洁区域然后将共聚焦环直接固定在要成像的皮肤上来准备该区域。施加超声凝胶并固定共聚焦探针。成像后,用过量的生理盐水清洁凝胶,然后如上重新施加测试样品(不具有诱饵分子的FITC-右旋糖酐),再如上放置2小时,之后重复成像流程。在不同深度(0、20、40和60微米)施加3hr后,进行共聚焦成像。图13所示的结果表明了在诱饵分子存在下,FITC-右旋糖酐在眉间区中的渗透。测试样品没有表现出任何渗透。如图14所示,FITC-右旋糖酐的渗透率随时间增加。图15显示了3hr后在多种诱饵分子ILS-20(A)和ILS-3(B)存在下,FITC-右旋糖酐在外侧眼角区域中的渗透。ILS-20是平均分子量为约20,000Da的透明质酸片段,ILS-3是平均分子量为约8,500Da的透明质酸片段。
实例25
肉毒杆菌毒素A的局部腋窝应用
方法.用肥皂和水清洁目标区域,彻底冲洗。重建2个100U 小瓶,每个小瓶含100uL等分试样的NS中的1%FITC-右旋糖酐(平均MW 150,000D)+1%多糖诱饵(ILS-20或ILS-3)。ILS-20是平均分子量为约20,000Da的透明质酸片段,ILS-3是平均分子量为约8,500Da的透明质酸片段。用注射器回收尽可能多的体积(未取下盖子;约80uL)。应用至曲腋处的腋中线,并在标准水肿男性受试者中轻轻按摩至每个腋窝。用Tegaderm敷料覆盖1小时,以使测试品的损失或扩散减到最小。去除敷料并用NS轻轻洗涤。在第10天,在引起出汗的标准化常规锻炼后,按照常规方法进行淀粉碘测试。结果示于图16-18中。
实例26
将多能干细胞的悬浮液注入功能有些减退的实体组织中。该悬浮液包括弹性蛋白片段,与没有弹性蛋白片段的注射液相比,该悬浮液允许细胞在整个组织中的渗透增加。几周后,该组织充满分化细胞,这些分化细胞足以逆转先前的潜在障碍并实现所需的组织功能。
实例27
将含有工程化细胞和长度很短的透明质酸片段的水凝胶应用于具有一些皮肤暴露区域的伤口。透明质酸链可使细胞在整个伤口中更快地渗透。与不存在透明质酸片段的情况相比,结果是更快的渗透、更均匀的分布和更高的保留。因此,伤口愈合得更快,并以最小的疤痕实现正常功能。
实例28
将微针施加到皮肤上,该微针在表面上具有工程化的自体细胞的局部悬浮液。该悬浮液含有浓度分别为0.5%和1.0%的弹性蛋白和透明质酸的低MW片段。由于存在这些寡肽/多肽和寡糖/多糖片段,因此与不存在这些片段的情况相比,细胞沉积得更快、在整个皮肤上的分布更均匀并且以更高速率保留。这导致皮肤表现出萎缩和衰老迹象减少。
实例29
将工程化的自体T细胞的悬浮液注射到含有残留癌细胞的实体组织中。该悬浮液包括弹性蛋白片段,与没有弹性蛋白片段的注射液相比,该悬浮液将允许细胞在整个组织中的渗透增加。几周后,组织将不再含有任何可存活的肿瘤细胞,并且该受试者将没有癌症。
实例30
抗体在完整皮肤中的诊断性渗透
具有高糖饮食和环境污染暴露的受试者将应用水凝胶,该水凝胶含有1%的寡糖和多糖片段(长度很短的透明质酸链)以及全长荧光标记抗体(MW高达160Kd)与最佳浓度的各种高级糖化终产物的混合物,并使凝胶静置3-6小时。然后清洁经处理的区域,并用UV光或滤光片拍照,以通过光学成像可视化抗体的强度和粒度。对照片进行定量评估以点画背景以上,这将反映确定抗原的存在。该受试者随后将重复监测后续干预的影响和生活方式的改变。
实例31
切除活检后抗体穿过组织屏障的诊断性渗透
具有Fitzpatrick 2型皮肤且先前有癌前病变切除史的受试者会出现高度可疑的黑色素瘤的病变。将进行边缘切除活检。将切除部位与含有寡糖和多糖片段(长度很短的透明质酸链)以及针对一种或多种黑色素瘤抗原的标记抗体的溶液一起温育30分钟,然后冲洗并可视化标记。有标记抗体迹象的区域将进一步切除,使得去除所有残留的微侵入。几年后,该受试者将与莫氏手术(Moh’s surgery)一起接受应用于新病变的相同方法,以确保去除细胞微侵入。
实例32
间质诊断贴剂
含有细胞外基质的肽基和糖基片段的混合物的水凝胶将被应用于受试者的肩膀。随后将应用离散的多结构域结合树脂贴剂并使其保持过夜,该贴剂可分别评估血清或间质液中的疏水组分、离子组分、小分子组分或蛋白质组分。随后将检查该贴剂以评估目的间质标志物和血清标志物。
实例33
紧急诊断
包含细胞外基质的肽基和/或糖基片段加d-二聚体反应性化合物(在d-二聚体存在下会产生颜色)的贴剂将应用于疑似患有肺栓塞同时从农村地区转至医疗护理的受试者。d-二聚体的存在将告知和指导即时和后续紧急护理。
实例34
透明质酸酶和利多卡因
使用含有透明质酸酶和诱饵分子的组合物评估利多卡因跨皮肤的递送。将制备含有1wt.%利多卡因、0.5wt%的5,000Da至10,000Da的诱饵分子(弹性蛋白片段)以及0.5wt.%的透明质酸酶。将获得可存活的猪皮肤并将其用于生产中真皮移植物。将移植物定位在透皮通量装置中。通量将持续12-20小时,并将样本应用并停留在供体皮肤表面上。洗涤并均质化来自每个池(每个室)的皮肤。使用光谱法测定澄清的匀浆溶液和流过样品的利多卡因含量。
实例35
弹性蛋白酶和米诺环素
如实例1所述,将使用含有弹性蛋白酶的组合物评估米诺环素跨皮肤的递送。体外测量米诺环素向猪皮肤中的递送,并与米诺环素从米诺环素在盐水中的组合物(即没有弹性蛋白酶和诱饵分子)中的递送进行比较。将制备含有1wt.%米诺环素、0.5wt%的5,000Da至10,000Da的诱饵分子(透明质酸片段)以及1wt.%的弹性蛋白酶的组合物。含有米诺环素和弹性蛋白酶的局部组合物可用于治疗或改善皮肤结构感染或失调,例如蜂窝组织炎。
实例36
弹性蛋白酶和维生素C
将制备含有维生素C、5,000Da至10,000Da的诱饵分子(透明质酸片段)和弹性蛋白酶的组合物。还将测试由盐水中的维生素C组成的对照配制品。将组合物置于Franz扩散池中,皮肤将扩散池的隔室分开。在固定时间后测量扩散池接收侧的维生素C浓度。将使用含有维生素C和弹性蛋白酶但不含诱饵分子的组合物重复此过程。
实例37
用于毛发生长的比马前列素的局部施用
将制备含有0.01wt.%比马前列素、约0.5wt.%的透明质酸酶和弹性蛋白酶的组合、以及0.5wt%的5,000Da至10,000Da的诱饵分子(透明质酸片段)的组合物。还将制备仅含有0.01wt.%比马前列素和仅含盐水的对照配制品。该组合物将应用于最近在大约21天前完成化疗周期并且经历接近全头皮脱发的受试者。相对于可比较的对照,预期用含有透明质酸酶/弹性蛋白酶的组合物治疗的受试者在1、2和4周时实现更快的毛发生长速率。另外,在用含有透明质酸酶/弹性蛋白酶和诱饵分子的组合物治疗的受试者中,预期头发的长度、厚度和密度更大。
实例38
用于干眼症的比马前列素
将制备含有0.01wt.%比马前列素、0.5wt%的5,000Da至10,000Da的诱饵分子(透明质酸片段)以及0.5wt.%的透明质酸酶的组合物。还将制备仅含有0.01%比马前列素和仅含盐水的对照配制品。这些组合物将以滴眼剂的形式施用于受试者的眼睛表面。相对于可比较的对照,预期用含有透明质酸酶的组合物治疗的受试者会从干眼症中恢复。
Claims (19)
1.一种组合物,其包含:
约0.001wt.%至约10wt.%的至少一种平均分子量为约2,000道尔顿至约20,000道尔顿的细胞外基质组分或其片段,和
活性剂。
2.如权利要求1所述的组合物,其中该组合物不包含任何可检测量的分子量超过60,000道尔顿的细胞外基质组分或其片段。
3.如权利要求1所述的组合物,其进一步包含约0.1wt.%至约10wt.%的酶,该酶选自由以下组成的组:透明质酸酶、弹性蛋白酶或其组合。
4.如权利要求1所述的组合物,其中该细胞外基质组分选自由以下组成的组:透明质酸、胶原蛋白、纤连蛋白、弹性蛋白、凝集素及其片段和其组合。
5.如权利要求1所述的组合物,其中该组合物包含约0.1wt.%至约25wt.%的活性剂。
6.如权利要求1所述的组合物,其中该活性剂选自由以下组成的组:小分子药物、大分子药物、生物制剂、抗体、嵌合抗体、抗体片段、诊断性抗体、抗原、肽、佐剂、抗氧化剂、化妆品成分、治疗性细胞、诊断剂、放射性示踪剂、造影剂、神经毒素、感觉改变剂及其组合。
7.一种向受试者递送活性剂的方法,该方法包括:
将组合物应用于该受试者的表面组织,该组合物包含约0.001wt.%至约10wt.%的平均分子量为约2,000道尔顿至约20,000道尔顿的细胞外基质组分或其片段、以及一种或多种活性剂。
8.如权利要求7所述的方法,其中该组合物不包含任何可检测量的分子量超过60,000道尔顿的细胞外基质组分或其片段。
9.如权利要求7所述的方法,其中该细胞外基质组分选自由以下组成的组:透明质酸、胶原蛋白、纤连蛋白、弹性蛋白、凝集素及其组合。
10.如权利要求7所述的方法,其中该组合物包含约0.1wt.%至约25wt.%的活性剂。
11.如权利要求7所述的方法,其中该活性剂选自由以下组成的组:小分子药物、大分子药物、生物制剂、抗体、嵌合抗体、抗体片段、诊断性抗体、抗原、肽、佐剂、抗氧化剂、化妆品成分、治疗性细胞、诊断剂、放射性示踪剂、造影剂、神经毒素、感觉改变剂及其组合。
12.如权利要求11所述的方法,其中所述治疗性细胞选自由以下组成的组:干细胞、基因工程化哺乳动物细胞、抗原呈递细胞及其组合。
13.一种治疗受试者的癌症的方法,该方法包括:
施用组合物,该组合物包含多个表达肿瘤抗原的治疗性细胞和约0.1wt.%至约10wt.%的细胞外基质组分、其片段及其组合。
14.如权利要求13所述的方法,其中该肿瘤抗原选自胃肿瘤、结肠肿瘤、前列腺肿瘤、宫颈肿瘤、皮肤肿瘤、子宫肿瘤、卵巢肿瘤、胰腺肿瘤、肾脏肿瘤、肝肿瘤、头颈部肿瘤、鳞状细胞肿瘤、胃肠道肿瘤、乳腺肿瘤、肺肿瘤和脑肿瘤。
15.一种治疗有需要的受试者的出汗过多的症状的方法,该方法包括:
将组合物施用于该受试者的表面组织,该组合物包含神经毒素剂和约0.001wt.%至约10wt.%的至少一种平均分子量为约2,000道尔顿至约20,000道尔顿的细胞外基质组分或其片段。
16.如权利要求15所述的方法,其中该组合物不包含任何可检测量的分子量超过60,000道尔顿的细胞外基质组分或其片段。
17.如权利要求15所述的方法,其中所述神经毒素剂选自由以下组成的组:A型肉毒杆菌毒素、B型肉毒杆菌毒素、C型肉毒杆菌毒素、D型肉毒杆菌毒素、E型肉毒杆菌毒素、F型肉毒杆菌毒素、abobotulinumtoxinA、BTX-A、daxibotulinumtoxinA、incobotulinumtoxinA、onabotulinumtoxinA、rimabotulinumtoxinB、突触融合蛋白、其衍生物、其药学上可接受的盐及其组合。
18.一种非侵入性地检测抗原的方法,该方法包括:
将透皮贴剂应用到表面组织上,该透皮贴剂包含组合物,该组合物包含抗体或抗体片段、以及约0.001wt.%至约10wt.%的至少一种平均分子量为约2,000道尔顿至约20,000道尔顿的细胞外基质组分或其片段;以及
通过成像检测该抗体或抗体片段。
19.如权利要求18所述的方法,其中该抗原是肿瘤抗原、癌症抗原、过敏原、细菌抗原、病毒抗原、药物、激素、植物凝集素、内毒素及其组合。
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US20190105261A1 (en) | 2019-04-11 |
EP3694489A2 (en) | 2020-08-19 |
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