CN116075314A - 用于治疗实体瘤和软瘤以及增殖性疾病的组合物和方法 - Google Patents
用于治疗实体瘤和软瘤以及增殖性疾病的组合物和方法 Download PDFInfo
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Abstract
本发明提供了一种在有需要的受试者中预防或治疗实体瘤和软瘤以及增殖性疾病的方法。所述方法包括向受试者施用有效量的植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包含黑种草、头状百里香、银斑百里香、牛至、百里香(Thymbraspicata)、百里香(Satujerathymbra)、芝麻和漆树,绞股蓝、乳香和人参的组,所述方法用于预防或治疗受试者的实体瘤和软瘤以及增殖性疾病。
Description
技术领域
本发明在一些实施例中,涉及用于治疗实体瘤和软瘤以及增殖性疾病的组合物和方法。
背景技术
“癌症”一词用于描述许多异常细胞分裂失控的疾病。癌性实体瘤和软瘤以及增殖性疾病最初可能出现在几乎身体的任何组织或器官中,并由于先天遗传因素和环境因素,例如某人的饮食或暴露于辐射、毒素等。尽管医学取得了进步,并且对癌性实体瘤和软瘤以及增殖性疾病的分子基础有了了解,但任何给定类型癌症的确切原因在很大程度上是未知的,尤其是在特定个体中。鉴于缺乏知识,很难找到有效治疗实体瘤和软瘤以及增殖性疾病的方法也就不足为奇了。
寻找有效的治疗方法也具有挑战性,因为癌症通常会对各种治疗策略产生抗药性。此外,鉴于某些类型的癌症从其原发性来源扩散的能力,治疗癌症的有效手段成为更大的挑战。这个过程称为转移,使肿瘤细胞能够通过血液和淋巴系统扩散到身体的其他重要部位。转移是难以开发有效癌症治疗的关键原因。
目前现有的癌症疗法包括多种不同的消融技术,例如外科手术;组织、超声波、射频和辐射的低温或加热方法;化学方法,如药物、细胞毒剂、单克隆抗体;或经动脉化学固定(TACE),以及它们的组合,依照基于所治疗癌症的特定类型和阶段的特定方案。然而,这些疗法与相当高的成本相关。此外,目前的治疗选择是高度侵入性的,与显著的毒性相关,并导致患者的整体生活质量较差。
标准的护理癌症肿瘤治疗通常将受影响组织的手术切除与化学疗法或放射疗法相结合。施用化疗剂的标准方法是通过血液,例如全身递送,可以通过各种途径例如静脉内和/或胃肠道递送来实现。然而,毒性是与全身递送的化学治疗药物相关的主要缺点。标准的护理手术治疗也带来了一些问题,包括癌细胞转移到血液和/或淋巴系统中,这导致癌细胞有机会转移到身体的其他部位并导致额外的肿瘤形成。
当无法进行手术时,对于实体瘤和软瘤以及增殖性疾病,可接受的治疗方法是使用放射或化学疗法。但与化疗或放疗前手术切除的肿瘤的存活率相比,无法手术的癌症的存活率较低。
某些癌症肿瘤类似于身体的组织,因此会削弱免疫系统识别和杀死它们的固有能力。几种抗癌技术(例如癌症疫苗)旨在刺激免疫系统对抗癌细胞。免疫系统对肿瘤细胞发动攻击的能力受到阻碍,因为肿瘤细胞几乎不表现为(如果有的话)该个体的外来抗原。此外,肿瘤中可能有许多不同类型的细胞。每种细胞类型都有不同的细胞表面抗原,再次阻止免疫系统的攻击。
根据疾病阶段,肿瘤可能过于晚期(例如,体积庞大),疫苗无法发挥作用。这些以及其他因素是为什么肿瘤可能缺乏足以刺激足够的免疫系统所需的足够数量的抗原(或靶标)。
一般而言,如果早期发现癌症,则针对癌症的标准治疗可能高度有效。然而,即使获得了最好的结果,这种治疗也是侵入性的、有毒的并且损害身体以及对患者的精神和情感要求很高。如果癌症在晚期发现,很少有治疗可以为患者提供长期生存的希望。
因此,本领域继续需要识别和开发在治疗实体瘤和软瘤以及增殖性疾病方面更有效并且总体上对个人和社会提供更低成本的新策略。
发明内容
根据本发明的一个方面,提供了一种在有需要的受试者中预防或治疗实体瘤和软瘤(soft tumor)以及增殖性疾病的方法,该方法包括向受试者施用有效量的植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分(fraction)、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物种选自一类含有黑种草(Nigella sativa)、头状百里香(Thymuscapitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbraspicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)、漆树(Rhus coriaria)、绞股蓝(Gynostemmapetaphyllum)、乳香和人参的组,所述方法用于预防或治疗受试者的实体瘤和软瘤以及增殖性疾病。
根据本发明的一个方面,提供了一种针对实体瘤和软瘤以及增殖性疾病的疫苗,包括有效量的的植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗或预防实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类含有黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)、漆树(Rhus coriaria)、绞股蓝(Gynostemmapetaphyllum)、乳香和人参的组。
根据本发明的一个方面,提供了一种药物组合物,包含有效量的植物种或其属衍生的组分,选择一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物的、其模拟物或其组合物的组,其中,所述组分能够治疗或预防实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类含有黑种草(Nigella sativa)、头状百里香(Thymuscapitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbraspicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)、漆树(Rhus coriaria)、绞股蓝(Gynostemmapetaphyllum)、乳香和人参的组,所述药物组合物用于预防或治疗实体瘤和软瘤以及增殖性疾病。
根据本发明的一个方面,提供了一种物质组合物,包含至少2种植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、合成类似物其、其模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类含有黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)、漆树(Rhus coriaria)、绞股蓝(Gynostemma petaphyllum)、乳香和人参的组。
根据本发明的一个方面,提供了一种食品补充剂,包含至少2种植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类含有黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)、漆树(Rhus coriaria)、绞股蓝(Gynostemma petaphyllum)、乳香和人参的组。
根据本发明的一个方面,所述方法、疫苗、药物组合物、组合物或食品补充剂包含菠萝蛋白酶或其类似物。
根据本发明的一个方面,所述方法、疫苗、药物组合物、组合物或食品补充剂包含菠萝提取物,其中,菠萝提取物包含菠萝蛋白酶或其类似物。
根据本发明的一个方面,所述方法、疫苗、药物组合物、组合物或食品补充剂包含含有色氨酸的植物提取物。
根据本发明的一个方面,提供了一种食品补充剂、组合物或提取物,进一步包括“贝都因茶(Beduin Tea)”,其中,所述“贝都因茶”包括玫瑰叶姜味草灌木(Rose LeavesMicromeria fruticose)、鼠尾草(Salvia)、柠檬草(cymbopgon)(柠檬醛)阿莱藤属(Aloysia)、马鞭草(verbena officinalis)、马郁兰(origanum majorana)、薄荷。
根据本发明的另一个方面,提供了一种食品补充剂、组合物或提取物,进一步包括“贝都因茶”,其中,所述“贝都因茶”包括百里香、鼠尾草、小豆蔻、肉桂、红茶、哈布克、马尔马亚。
本发明所述百里香成分的更多细节包含在附录1中。
根据本发明的一些实施例,所述实体瘤和软瘤以及增殖性疾病选自一类含有肉瘤和癌的组,例如纤维肉瘤、粘液肉瘤、脂肪肉瘤、软骨肉瘤、成骨肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤,尤文氏瘤,平滑肌肉瘤,横纹肌肉瘤,结肠癌,胰腺癌,乳腺癌,卵巢癌,前列腺癌,鳞状细胞癌,基底细胞癌,腺癌,汗腺癌,皮脂腺癌,乳头状癌,乳头状腺癌,囊腺癌,髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎癌、维尔姆瘤、宫颈癌、睾丸肿瘤、肺癌、小细胞肺癌、膀胱癌、上皮癌、多形性胶质母细胞瘤、星形细胞瘤、髓母细胞瘤、颅咽管瘤、室管膜瘤、松果体瘤、血管母细胞瘤、听神经瘤、少突胶质细胞瘤、皮肤T细胞淋巴瘤(CTCL)、黑色素瘤、神经母细胞瘤和视网膜母细胞瘤。
根据本发明的一些实施例,所述实体瘤、软瘤以及增殖性疾病是脑癌、乳腺癌、三阴性乳腺癌、膀胱癌、骨癌、结肠直肠癌、肺癌、肾癌、肝癌、胃癌、前列腺癌癌症、肉瘤、黑色素瘤、癌、或淋巴瘤。
根据本发明的具体实施例,所述实体瘤和软瘤以及增殖性疾病是前列腺癌、乳腺癌、结肠直肠癌、胰腺癌、或淋巴瘤。
根据本发明的一些实施例,所述实体瘤和软瘤以及增殖性疾病是淋巴瘤。
根据本发明的一些实施例,所述增殖性疾病是纤维瘤(Fibroids)。
根据本发明的一些实施例,所述增殖性疾病是子宫内膜异位症。
根据本发明的一些实施例,所述组分包括至少2个组分。
根据本发明的一些实施例,所述组分包括至少3个组分。
根据本发明的一些实施例,所述组分包括至少4个组分。
根据本发明的一些实施例,所述组分包括至少5个组分。
根据本发明的一些实施例,所述组分包括5-10个组分。
根据本发明的一些实施例,所述组分包含百里醌或其类似物。
根据本发明的一些实施例,所述组分包括百里酚或其类似物。
根据本发明的一些实施例,所述组分包括香芹酚或其类似物。
除非另有定义,否则本发明使用的所有技术和/或科学术语具有与本发明所属领域的普通技术人员通常理解的相同含义。尽管与本发明所述的方法和材料相似或等效的方法和材料可以用于本发明的实施例的实践或测试,但示例性方法和/或材料如下文描述。如有冲突,以专利说明书(包括定义)为准。此外,这些材料、方法和示例仅是说明性的,并不意味着必然是限制性的
附图说明
本发明仅通过示例的方式参考附图描述了本发明的一些实施例。现在详细地具体参考附图,应强调的是,所示的细节仅作为示例并且出于说明性讨论本发明的实施例的目的。就这一点而言,本领域技术人员结合附图的描述能够显而意见的实施本发明的实施例。
图1A-C显示了取自berkem(dot)com的植物提取方法的实施例。图1A-描述植物提取一般原理的示意图;图1B-描述根据一些实施例的主要分离过程的示意图;图1C-描述可能影响过程的参数的示意图。
图2描绘了一名疑似患有基底细胞癌的鳞状细胞癌患者在治疗前的变色和色素沉着,日期为2020年7月28日。
图3描绘了2020年10月30日治疗后疑似疑患有基底细胞癌的鳞状细胞癌患者的变色和色素沉着。
具体实施方式
在本发明在其一些实施例中,涉及用于治疗和预防实体瘤和软瘤以及增殖性疾病的组合物和方法。
在详细解释本发明的至少一个实施例之前,应当理解,本发明在其应用中不必限于以下描述中阐述的或由实施例所例示的细节。本发明能够具有其他实施例或能够以各种方式实践或执行。
本发明的靶组织为实体瘤和软瘤以及增殖性疾病,特别是恶性实体瘤和软瘤以及增殖性疾病。本发明提供了用于治疗实体瘤和软瘤以及增殖性疾病的基于植物的组合物和/或组分。实体瘤和软瘤以及增殖性疾病是指组织异常肿块,通常不包含囊肿或液体区域。本发明的基于植物的组合物或组分,当施用于患有实体瘤和软瘤以及增殖性疾病的受试者时可以具有治疗效果(仅举几例,组合物和/或组分可以减轻实体瘤和软瘤以及增殖性疾病的症状,导致肿瘤块消退,减缓癌症进展或治愈癌症)。目前,对于许多形式的实体瘤和软瘤以及增殖性疾病及其伴随症状,还没有有效的预防性治疗。
因此,根据本发明的一个方面,提供了一种在有需要的受试者中治疗实体瘤和软瘤以及增殖性疾病的方法,所述方法包括向受试者施用有效量的植物物种或属衍生的组分,选自一类含有植物部分、其提取物、其级分(fraction)、其活性成分、其合成类似物、其模拟物或其组合物的组,其中所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)和漆树(Rhus coriaria)、绞股蓝(Gynostemma petaphyllum)、乳香和人参的组,所述方法用于预防或治疗受试者的实体瘤和软瘤以及增殖性疾病。
根据本发明的替代或附加的方面,提供了一种针对实体瘤和软瘤以及增殖性疾病生长的疫苗,所述疫苗包含有效量的植物物种或属衍生的组分,选自一类含有植物部分、其提取物、其级分(fraction)、其活性成分、其合成类似物、其模拟物或其组合物的组,其中所述组分能够治疗和/或减缓实体瘤和软瘤以及增殖性疾病的生长,并且其中所述植物物种选自一类包括黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)和漆树(Rhus coriaria)、绞股蓝(Gynostemma petaphyllum)、乳香和人参的组。
根据本发明的替代或附加的方面,提供了一种药物组合物,其包含有效量的植物物种或属衍生的组分,选自一类含有植物部分、其提取物、其级分(fraction)、其活性成分、其合成类似物、其模拟物或其组合物的组,其中所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草(Nigella sativa)、头状百里香(Thymus capitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbra spicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)和漆树(Rhuscoriaria)、绞股蓝(Gynostemmapetaphyllum)、乳香和人参的组,所述药物组合物用于治疗实体瘤和软瘤以及增殖性疾病。
根据本发明的替代或附加方面,提供了一种物质组合物,其包含至少2种植物物种或属衍生的组分,选自一类含有植物部分、其提取物、其级分(fraction)、其活性成分、其合成类似物、其模拟物或其组合物的组,其中所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草(Nigella sativa)、头状百里香(Thymuscapitatus)、银斑百里香(Thymus vulgaris)、牛至(Origanum syriacum)、百里香(Thymbraspicata)、百里香(Satujera thymbra)、芝麻(Sesamum indicum)和漆树(Rhus coriaria)、绞股蓝(Gynostemmapetaphyllum)、乳香和人参的组。
根据本发明的实施例,所述方法、疫苗、药物组合物、组合物或食品补充剂包含菠萝蛋白酶或其类似物。
根据本发明的实施例,所述方法、疫苗、药物组合物、组合物或食品补充剂包含菠萝提取物,其中所述菠萝提取物包含菠萝蛋白酶或其类似物。
本发明承认本发明的各方面提供了用于治疗或预防肿瘤病毒(Oncoviruses)的组合物和方法。
肿瘤病毒:
肿瘤病毒或致癌病毒是一种可导致癌症的病毒。[4]本发明中的术语肿瘤病毒是指任何具有DNA或RNA基因组的导致癌症的病毒,并且与“肿瘤病毒”或“癌症病毒”同义。
卡波西肉瘤(Kaposi's sarcoma)是一种可在皮肤中形成肿块的癌症,由卡波西肉 瘤相关疱疹病毒(KSHV),也称为HHV-8引起。
DNA病毒人乳头瘤病毒(HPV)是一种DNA病毒,通过干扰肿瘤抑制蛋白例如p53引起细胞转化。干扰p53的作用可使感染病毒的细胞进入细胞周期的不同阶段,使病毒基因组得以复制。迫使细胞进入细胞周期的S期可能会导致细胞发生转化。[25]人乳头瘤病毒感染是导致宫颈癌、外阴癌、阴道癌、阴茎癌、肛门癌和HPV-阳性口咽癌的主要原因。[7][26][27][28][29][30][31]有近200种不同的人乳头瘤病毒(HPVs),[29]许多HPV类型是致癌的。[7][26]
卡波西肉瘤相关疱疹病毒(KSHV或HHV-8)与卡波西肉瘤(一种皮肤癌)有关。[32]
人类疱疹病毒第四型(Epstein–Barr virus)(EBV或HHV-4)与四种癌症有关。
默克尔细胞多瘤病毒(Merkel cell polyomavirus)-一种多瘤病毒-与默克尔细 胞癌的发展有关。[24]
人类巨细胞病毒(CMV或HHV-5)与粘液表皮样癌和其他可能的恶性肿瘤有关。[33]
RNA病毒
一些RNA病毒也与癌症有关,例如丙型肝炎病毒以及某些逆转录病毒,例如人类T 淋巴细胞病毒(HTLV-1)和劳斯肉瘤病毒(Rous sarcoma virus)(RSV)。
如本发明所用,术语“植物”包括整株植物、嫁接植物、植物的祖先和后代以及植物部分,包括种子、花、树皮、芽、茎、根(包括块茎)、果实、根茎、接穗、以及植物细胞、组织和器官。
根据一个具体实施例,所述植物部分是种子。
根据一个具体实施例,所述植物部分是果实。
根据一个具体实施例,所述植物部分是叶。
根据一个具体实施例,所述植物部分是茎。
根据一个具体实施例,所述植物部分是花。
植物部分可以是固体部分或非固体部分,例如植物的油或含水部分。
植物可以是任何形式,包括悬浮培养物、胚胎、分生组织区、愈伤组织、叶、配子体、孢子体、花粉和小孢子。
术语植物是指野生植物或其栽培品种。
如本发明所用,术语“植物物种”是指该属内的一种或多种植物的亚组。这些植物将彼此共享相似的特征。一个物种中可能只有一种植物,也可能有数百种植物。该术语旨在包括亚种,例如不同地理位置生长或可被发现的亚种,例如黎巴嫩漆树和叙利亚漆树或高丽参和西洋参。
如本发明所用,“植物属”是指低于科和高于物种的分类等级。
应当理解,下面列出的相关种和属以及它们的每个选项或其组合代表本发明的不同实施例。
术语“提取”是指一种分离过程,该过程依赖于一种或多种分析物与一种或多种分析物以外的样品组分的分离。提取是通常使用两个不混溶的相以将一种或多种溶质从一相进入另一相进行分离的过程。溶质在两相之间的分布是分配理论描述的平衡条件。例如,在水中煮沸茶叶会将单宁、可可碱和咖啡因从叶子中提取到水中。更典型的通常不仅是在实验室中进行的提取而是将有机化合物从水相中提取到有机相中。常见的提取剂根据Hildebrand溶解度参数按极性升序从乙酸乙酯到水排列(乙酸乙酯<丙酮<乙醇<甲醇<丙酮:水(7:3)<乙醇:水(8:2)<甲醇:水(8:2)<水)。植物提取过程见图1A-C。
如本发明所用,术语“提取物”是指这种分离过程的结果,其可以根据提取过程采取溶液制剂或其他化学形式的形式。特别地,术语提取物可以涉及通过提取样品(例如原材料)的一部分制成的物质,例如通过使用诸如乙醇或水的溶剂。在各种情况下,提取物涉及富含一种或多种溶质的溶剂。特别地,在本发明公开的意义上的“植物提取物”通常包括植物材料的浓缩制剂,其通过用一种或多种提取方法分离或纯化所需的活性成分而获得。
溶剂的选择取决于要获得的所需组分。例如,为了在提取过程中提取极性组分,建议的溶剂包括但不限于水、乙醇、甲醇或丁醇,而对于非极性化合物,根据提取物的用途,使用乙醚、己烷或氯仿。对于中极性,可以选择乙酸乙酯,但也可以使用其他溶剂。
固/液萃取的一般程序可以按五种不同的方式进行缩放:
浸渍:接触阶段保持在室温。
煎煮或回流:接触阶段保持在溶剂的沸点。
消化:接触阶段保持在前两种情况之间的温度
灌注:将沸腾的溶剂倒在固体上,然后冷却一段时间。
浸出或渗滤:溶剂通过生物质。
也可以将这些方法相互结合或与其他工艺如蒸馏、蒸汽蒸馏、精馏等结合。
根据另一个实施例,考虑连续或组合使用各种溶剂,并且有机化学的普通技术人员将知道根据如下所述的活性成分选择哪一种。
可以通过其他方式进一步辅助提取,例如超滤、反渗透、高压(超临界CO2)、微波、超声波等。
在一些实施例中,植物部分与极性溶剂(例如乙醇)或非极性溶剂(例如己烷或戊烷)接触几分钟,例如15分钟或更长、约30分钟或更长、约1小时或更长,约2小时或更长,或约5小时或更长。
在接触期间也可以控制温度。
根据具体实施例,植物部分与溶剂(例如乙醇)接触,同时不断混合,例如在振动筛上。
应当理解,提取过程也可以是无溶剂的。
例如,无溶剂微波萃取(SFME)已被提议作为一种从广泛用于食品工业的芳香草本植物中提取精油的绿色方法。该技术结合了微波加热和在大气压力下进行的干馏,无需添加任何溶剂或水。挥发性化合物的分离和浓缩在一个阶段进行。在一些实施例中,SFME和/或水蒸馏(HD)用于从本发明的植物中提取精油。
在一些实施例中,本发明的过程包括从包含液体提取物和固体的混合物(即粗提物)中分离液体提取物(即过滤的提取物)。用于分离液体提取物(即过滤的提取物)的合适方法包括有机合成领域中已知的那些并且包括但不限于重力过滤、抽吸和/或真空过滤、离心、凝结和倾析等。在一些实施例中,分离包括通过孔径为约1-5μm、约0.5-5μm、约0.1-5μm、约1-2μm、约0.5-2μm、约0.1-2μm、约0.5-1μm、约0.1-1μm、约0.25-0.45μm、或约0.1-0.5μm(例如约2μm、约1μm、约0.45μm或约0.25μm)的多孔膜、注射器、海绵、沸石、纸等过滤液体提取物。
根据具体实施例,本发明考虑对生成后的过滤提取物进行干燥(即去除极性/非-极性溶剂)和/或冷冻。
干燥过滤的提取物(即去除极性溶剂)的方法没有特别限制,并且可以包括在减压(例如,低于大气压)和/或升高的温度(例如,高于约25℃)下蒸发溶剂。在一些实施例中,可能难以通过标准溶剂去除程序从液体提取物中完全去除溶剂,例如蒸发。在一些实施例中,如共蒸发、冻干等过程可用于从液体级分中完全去除极性溶剂以形成干粉、干丸、干颗粒、糊剂等。根据一个具体实施例,极性溶剂用真空蒸发器蒸发。
提取过程的选择很大程度上取决于要分离的成分。
应当理解,在提取物的产生之后,本发明的具体实施例进一步考虑了附加的纯化步骤,以便进一步从提取物中分离/纯化活性剂,例如,将过滤后的提取物分级。
如本发明所用,“级分”是指仅包含提取物的某些化学成分但不是全部的提取物的一部分。
分级(fractionating)可以通过例如但不限于柱色谱法、制备型高效液相色谱法(“HPLC”)、减压蒸馏及其组合的方法进行。
根据一个具体实施例,分级通过HPLC进行。
在一些实施例中,分级包括将过滤的提取物重新悬浮在极性溶剂(如甲醇,如上所述)中,将极性提取物施加到分离柱,并通过柱色谱法分离具有抗呼吸系统疾病(例如抗纤维化、抗炎)活性的提取物(制备型高效液相色谱法)。
将洗脱溶剂应用于分离极性提取物的分离柱,以从极性提取物中洗脱级分(fractions)。适用的洗脱溶剂包括但不限于甲醇、乙醇、丙醇、丙酮、乙酸、甲乙酮、乙腈、丁腈、二氧化碳、乙酸乙酯、四氢呋喃、二异丙醚、氨、三乙胺、N,N-二甲基甲酰胺、N,N-二甲基乙酰胺等及其组合。
根据替代或附加的实施例,液相色谱法包括高效液相色谱法(HPLC)。
根据替代或附加的实施例,液相色谱在反向固定相上进行。
级分可以通过分析方法表征,例如但不限于光谱方法,例如但不限于紫外-可见光谱(“UV-Vis”)、红外光谱(“IR”)等;质谱(“MS”)方法,例如但不限于飞行时间质谱;四极质谱;电喷雾质谱、傅里叶变换质谱、基质辅助激光解吸/电离(“MALDI”)等;色谱方法,例如但不限于气相色谱(“GC”)、液相色谱(“LC”)、高效液相色谱(“HPLC”)等;及其组合(例如,GC/MS、LC/MS、HPLC/UV-Vis等),以及本领域普通技术人员已知的其他分析方法。
获得的组分(活性成分、提取物和/或级分)可以用治疗实体瘤和软瘤以及增殖性疾病或其症状来检测。用于测试效果的示例性方法在下文以及随后的实施例部分中进一步描述。
本发明所述的活性成分、提取物和/或级分可以立即使用或储存直至进一步使用。
根据具体实施例,活性成分、提取物和/或级分在冷冻下保存需要的时间长度,例如冷冻柜中,直到进一步使用(例如在大约-20℃到-90℃,在大约-70℃到-90℃,例如在-80℃)。
根据其他具体实施例,立即使用活性成分、提取物和/或级分(例如在几分钟内,例如最多30分钟)。
活性成分、提取物和/或级分可以单独使用。可替代地,可以将不同的活性成分、提取物和/或级分(例如来自不同植物或来自单独的提取程序)合并在一起。同样,不同的活性成分、提取物和/或级分(来自相同的提取物、来自不同的提取物、来自不同的植物和/或来自不同的提取程序)可以合并在一起。
使用本教导,本发明人不仅能够鉴定出可用于有效治疗或预防实体瘤和软瘤以及增殖性疾病的植物和提取物,而且能够鉴定出其活性成分。
“活性成分”是指对实体瘤和软瘤以及增殖性疾病具有抗(预防或治疗)效果的确定的化学成分。
活性成分可以从植物中提纯或化学合成(人工、人造)。
本发明还考虑了活性成分的类似物和衍生物,只要维持抗实体瘤和软瘤以及增殖性疾病的抗肿瘤(预防或治疗)效果(参见例如以下实施例部分),它们也称为模拟物。
以下是从本发明的选定植物中提取活性成分的一些非限制性实施例。
从头状婆罗门参(T.capitatus)的叶子中提取-收集头状婆罗门参(叶子)样品的地上部分。从树枝上分离出来的叶子在室温下脱水7天,然后稍微混合成细粉进行提取。
精油(EO)提取-水蒸馏用于从植物中提取EO,例如头状婆罗门参的干燥地上部分。简而言之,通过将100g植物与500mL蒸馏水混合,进行数小时(例如3小时)的提取。将提取物干燥并使用硫酸钠和旋转蒸发器在减压下浓缩。EO产量是根据每100g干燥植物获得的油量(以mL为单位)确定的。最后,纯EO储存在-4℃直至进一步分析。
精油分析-EO的化学成分通过GC和GC-MS进行检查。GC分析使用气相色谱仪进行。成分的比例由峰面积的积分决定。此外,质谱法(MS)通常可用于在与上述气相色谱法相同的条件下分析EO。基于质谱的拟合和纯度,通过将它们的保留指数(相对于一系列正烷烃的保留时间确定)与Wiley库搜索例程12的标准的保留指数进行比较来确定不同化合物的鉴定。这样的条件用于确定如下所述的活性成分。
从百里香(Satujera Thymbra)中提取:
2009年4月在黎巴嫩随机收集了来自S.thymbra的风干地上部分。使用克莱文格式装置将植物材料进行蒸汽蒸馏3小时以生产精油,产量为0.84%(w/w)。使用无水硫酸镁干燥油并在4℃下储存。通过GC/MS分析S.thymbra油。
从漆树(Sumac)中提取
为了从漆树果实中分离、测定和鉴定化合物,从漆树植物的果实或叶子中提取不同的提取物。有些是从水提取物中分离出来的,有些是从酒精提取物中分离出来的,有些是从脂质提取物中分离出来的。可水解单宁在漆树果实中的比例最高,其次是黄酮类化合物。这强调了水果的抗氧化潜力。在可水解单宁之后,占水果质量近20%的是其他未鉴定的化合物。随后有花青素、异黄酮、萜类和二萜类。对漆树成熟果实进行化学性质分析,发现其蛋白质含量为2.6%,脂肪含量为7.4%,纤维含量为14.6%,灰分含量为1.8%。此外,热量计算表明100克漆树果实含有147.8kcal。
从黑种草中提取百里醌
可以使用各种方法,包括具有温度控制功能的微波辅助提取系统以及其他提取方法、索氏提取法和常规的固/液提取。
黑种草
根据一个具体实施例,活性成分(例如,可以通过超临界二氧化碳萃取法获得)包括但不限于:
本发明考虑的其他植物是黑种草属。
黑种草是毛茛科18种一年生植物的一个属,原产于南欧、北非、南亚、西南亚和中东。 应用于该属成员的常用名称是黑种草、灌木丛中的恶魔或迷雾中的爱。
Nigella arvensis
Nigella carpatha
Nigella damascena
Nigella degenii
Nigella deserti
Nigella doerfleri
Nigella elata
Nigellafumariifola
Nigella hispanica
Nigella latisecta
Nigella nigellastrum
Nigella orientalis
Nigella oxypetala
thymo
Nigella sativa
Nigella segetalis
Nigella stricta
Nigella unguicularis
根据一个具体实施例,所述活性成分是百里醌。
其他含有百里醌的植物包括但不限于:
Monarda fistulos(美国薄荷属);
Satureja montana(香薄荷属);
其他含有百里醌的家族包括但不限于:
菊科-示例包括但不限于以下亚科:
·刺菊木亚科Bremer&Jansen
·管状花亚科Sweet
·菊苣亚科Chevallier
·伞形科Panero&Funk
·Famatinanthoideae S.E.Freire,Ariza&Panero
·Gochnatioideae Panero&Funk
·Gymnarrhenoideae Panero&Funk
·Hecastocleidoideae Panero&Funk
·Mutisioideae Lindley
·Pertyoideae Panero&Funk
·Stifftioideae Panero
·Wunderlichioideae Panero&Funk
Cupressaceae柏科
________________________________________________________
·Cunninghamioideae杉亚科
·Taiwanioideae台湾杉亚科
·Athrotaxidoideae密叶杉亚科
·Sequoioideae红杉亚科
·Taxodioideae落羽杉亚科
·Callitroideae柏松亚科
·Cupressoideae柏亚科
·Incertae sedis
Lamiacea唇形科
Ranunculacea毛茛科
·Hydrastidoideae海蜇科
·Glaucidioideae海藻科
·Coptoideae
·Thalictroideae唐松草亚科
·Ranunculoideae毛茛亚科
含有香芹酚的植物清单包括但不限于:
Monarda didyma
Nigella sativa
Origanum compactum
Origanum dictamnus
Origanum microphyllum
Origanum onites
Origanum scabrum
Origanum syriacum
Origanum vulgare
Plectranthus amboinicus
Thymus glandulosus
Lavandula multifida
Origanum minutiflorum
Satureja thymbra
头状百里香(Thymus Capitatus)中发现的活性成分
本发明考虑的其他植物是百里香属植物。
茎趋于狭窄或甚至呈丝状;在大多数物种中,叶子是常绿的,成对排列,椭圆形,全叶,小,长4-20毫米,通常具有芳香味。百里香花在密集的顶生头状花序中,花萼不均匀,上唇三裂,有黄色、白色或紫色。
该属的几个成员被种植为烹饪草本或观赏植物,当它们也被称为百里香时,以其最著名的品种,银斑百里香(Thymus vulgaris)或普通百里香命名。
约350种,包括:
Thymus adamovicii
Thymus altaicus
Thymus amurensis
Thymus boissieri
Thymus bracteosus
Thymus broussonetii
Thymus caespititius
Thymus camphoratus
Thymus capitatus
Thymus capitellatus
Thymus camphoratus
Thymus carnosus
Thymus cephalotus
Thymus cherlerioides
Thymus ciliatus
Thymus cilicicus
Thymus cimicinus
Thymus citriodorus(Thymus×citriodorus)同义词T.fragrantissimus,T.serpyllum citratus,T.serpyllum citriodorum.[7]–柑橘百里香
Thymus comosus
Thymus comptus
Thymus curtus
Thymus decussatus
Thymus disjunctus
Thymus doerfleri
Thymusglabrescens
Thymus herba-barona
Thymus hirsutus
Thymus hyemalis
Thymus inaequalis
Thymus integerThymus lanuginosus,同义词T.serpyllum–羊毛百里香Thymusleucospermus
Thymus leucotrichus
Thymus longicaulis
Thymus longiflorus
Thymus mandschuricus
Thymus marschallianus
Thymus mastichina
Thymus membranaceus
Thymus mongolicus
Thymus moroderi
Thymus nervulosus
Thymus nummularis
Thymus odoratissimus
Thymus pallasianus
Thymus pallidus
Thymus pannonicus
Thymus praecox–匍匐百里香
Thymus proximus
Thymus pseudolanuginosus,同义词T.serpyllum–羊毛百里香
Thymus pulegioides–柠檬百里香[8]
Thymus quinquecostatus
Thymus richardii
Thymus satureioides
Thymus serpyllum
Thymus sibthorpii
Thymus striatus
Thymus thracicus–薰衣草百里香
Thymus villosus
Thymus vulgaris–普通百里香
Thymus zygis
含有百里酚的植物清单包括但不限于:
Euphrasia rostkoviana
Lagoecia cuminoides
Monarda didyma
Monarda fistulosa
Mosla chinensis,Xiang Ru
Origanum compactum
Origanum dictamnus
Origanum onites
Origanum vulgare
Satureja thymbra
Thymus glandulosus
Thymus hyemalis
Thymus vulgaris
Thymus zygis
Trachyspermum ammi
银斑百里香中的活性成分:
根据本发明的一些实施例,百里香的精油(EO)上的活性成分包括但不限于:
百里香(Satujera Thymbra)的活性成分:
2009年4月在黎巴嫩随机收集了来自S.thymbra的风干地上部分。使用克莱文格(clevenger)式装置将植物材料进行蒸汽蒸馏3小时以生产精油,产量为0.84%(w/w)。油用无水硫酸镁干燥并储存在4℃。通过GC/MS分析S.thymbra油。鉴定出占油样98.8%的19种化合物。Satureja thymbra L.油的主要成分是γ-松油烯(34.06%)、香芹酚(23.07%)和百里酚(18.82%)。同样丰富的还有对伞花烃(7.58%)、石竹烯(3.96%)、α-松油烯(3.53%)和月桂烯(1.70%)。
本发明还考虑了夏香草(Satujera)属的植物。
夏香草(Satureja)是唇形科的芳香植物属,与迷迭香和百里香有关。它原产于北非、欧洲南部和东南部、中东和中亚。一些新世界物种以前包括在Satureja中,但它们都已移至其他属。有几个物种被种植为称为咸味的烹饪草药,它们已经在一些地方的野外扎根。
示例包括但不限于:
Satureja aintabensis P.H.Davis-土耳其
Satureja amani P.H.Davis-土耳其
Satureja atropatana Bunge-伊朗
Satureja avromanica Maroofi-伊朗
Satureja bachtiarica Bunge-伊朗
Satureja boissieri Hausskn.ex Boiss.-土耳其、伊朗
Satureja bzybica Woronow-高加索地区
Satureja×caroli-paui G.López-西班牙(S.innota×S.montana)
Satureja cilicica P.H.Davis-土耳其
Satureja coerulea Janka-保加利亚、罗马尼亚、土耳其
Satureja cuneifolia Ten-西班牙、意大利、希腊、阿尔巴尼亚、南斯拉夫、伊拉克
Satureja×delpozoi Sánchez-Gómez,J.F.Jiménez&R.Morales-西班牙(S.cuneifolia×S.intricata var.gracilis)
Satureja edmondii Briq.-伊朗
Satureja×exspectata G.López-西班牙(S.intricata var.gracilis×S.montana)
Satureja hellenica Halácsy-希腊
Satureja hortensis L.
Satureja icarica P.H.Davis-希腊群岛
Satureja innota(Pau)Font Quer-西班牙
Satureja intermedia C.A.Mey.-伊朗、高加索地区
Satureja intricata Lange-西班牙
Satureja isophylla Rech.f.-伊朗
Satureja kallarica Jamzad-伊朗
Satureja kermanshahensis Jamzad-伊朗
Satureja khuzistanica Jamzad-伊朗
Satureja kitaibelii Wierzb.ex Heuff.-保加利亚、罗马尼亚、南斯拉夫
Satureja laxiflora K.Koch-伊朗、伊拉克、土耳其、高加索地区
Satureja linearifolia(Brullo&Furnari)Greuter-利比亚昔兰尼加地区
Satureja macrantha C.A.Mey.-伊朗、伊拉克、土耳其、高加索地区
Satureja metastasiantha Rech.f.-伊拉克
Satureja montana L.–winter savory-南欧、土耳其、叙利亚
Satureja mutica Fisch.&C.A.Mey.-高加索地区、伊朗、土库曼斯坦
Satureja nabateorum Danin&Hedge-约旦
Satureja pallaryi J.Thiébaut-叙利亚
Satureja parnassica Heldr.&Sart.exBoiss.-希腊、土耳其
Satureja pilosa Velen.-意大利、希腊、保加利亚
Satureja rumelica”Velen.-保加利亚
Satureja sahendica Bornm.-伊朗
Satureja salzmannii(Kuntze)P.W.Ball-摩洛哥、西班牙
Satureja spicigera(K.Koch)Boiss.-土耳其、伊朗、高加索地区
Satureja spinosa L.-土耳其、希腊群岛,包括克里特岛
Satureja subspicata Bartl.ex Vis.-奥地利、南斯拉夫、阿尔巴尼亚、保加利亚、意大利
Satureja taurica Velen.-克里米亚
Satureja thymbra L.-利比亚,欧洲东南部,从撒丁岛到土耳其;塞浦路斯、黎巴嫩、巴勒斯坦
Satureja thymbrifolia Hedge&Feinbrun-以色列、沙特阿拉伯
Satureja wiedemanniana(Avé-Lall.)Velen.–土耳其
百里香(Thymbra spicata)的活性成分:
1RT-保留时间;2Rl-保留指数;3萘,1,2,3,4,4a,5,6,7-八氢-4a-甲基
本发明还考虑的是百里香(Thymbra)属植物。
Thymbra,俗称地中海百里香,是唇形科植物的一个属。按照目前的分类,该属有七个种和一个亚种。它原产于南欧、北非和中东的地中海地区。
示例包括但不限于:
Thymbra calostachya(Rech.f.)Rech.f.-克里特岛
Thymbra capitata(L.)Cav.-从摩洛哥+葡萄牙到土耳其+巴勒斯坦广泛分布
Thymbra sintenisii Bornm.&Azn.-伊拉克、土耳其
Thymbra spicata L.-希腊、土耳其、叙利亚、黎巴嫩、巴勒斯坦、以色列、伊拉克、伊朗
西西里漆树(漆树)的化学成分
使用HPLC-MS方法表征和鉴定漆树的化学成分,鉴定出西西里漆树中的191种化合物,并将它们分类为:
·78种可水解单宁(例如没食子单宁,例如五、六、七、八、九和十没食子酰葡萄糖苷)
·59种类黄酮化合物(例如,槲皮素、杨梅素3-鼠李糖苷和槲皮素3-葡萄糖苷)
·9种花青素(例如飞燕草素-3-葡萄糖苷(Delphidin-3-glucoside)、花青素3-(2"-没食子酰基)半乳糖苷(Cyanidin 3-(2"-galloyl)galactoside)、花青素-3-葡萄糖苷(Cyanidin-3-glucoside)、7-甲基-花青素-3-(2"没食子酰基)半乳糖苷(7-methyl-cyanidin-3-(2"galloyl)galactoside)、7-甲基-花青素-3-半乳糖苷(7-methyl-cyanidin-3-galactoside))
·2种异黄酮
·2种萜类
·1种二萜类
·38种其他未鉴定出的化合物。
根据具体实施例,所述漆树中的酚类化合物是与花青素一起构成其植物化学活性的化合物。发现漆树果实中最丰富的酚类化合物是没食子酸。
可水解单宁在漆树果实中的百分比最高,其次是类黄酮。这强调了在本发明中作为具体实施例考虑的植物部分果实的抗氧化潜力。在可水解单宁之后,其他未鉴定出的化合物占水果质量的近20%。随后有花青素、异黄酮、萜类和二萜类。漆树果实的化学性质是在成熟果实上进行的,发现其蛋白质含量为2.6%,脂肪含量为7.4%,纤维含量为14.6%,灰分(ash)为1.8%。此外,热量计算表明100克漆树果实含有147.8kcal。
可水解单宁在漆树果实中的百分比最高,其次是类黄酮。这强调了在本发明中果实的抗氧化潜力。在可水解单宁之后,其他未鉴定出的化合物占水果质量的近20%。随后有花青素、异黄酮、萜类和二萜类。漆树果实的化学性质是在成熟果实上进行的,发现其蛋白质含量为2.6%,脂肪含量为7.4%,纤维含量为14.6%,灰分(ash)为1.8%。此外,热量计算表明100克漆树果实含有147.8kcal。
其他活性成分或其任何组合物包括但不限于没食子酸甲酯、茄子黄酮(gathisflavone)、舒黄酮(sumaflavone)、欣菲克黄酮(hinfikflavone)、光儿茶酸、五没食子酰葡萄糖、扁柏黄酮、β-石竹烯、飞燕草素-3-葡萄糖苷、花青素3-(2"-没食子酰)半乳糖苷、花青素-3-葡萄糖苷、7-甲基-花青素-3-(2"没食子酰基)半乳糖苷、7-甲基-花青素-3-半乳糖苷、槲皮素-3-葡萄糖苷、山柰酚(kampferol)、杨梅素、紫铆花素、D-柠檬苦素。
根据一个具体实施例,所述活性成分或其组合物包括挥发性化合物,例如萜烯烃、单萜烃和倍半萜烃,特别是β-石竹烯和α-蒎烯、芫荽(Coririanaphthyl)醚、芫荽(Coriarioic)酸和芫荽(Coriariaacthracenyl)酯。
根据一个具体实施例,所述活性成分或其组合物包括脂肪酸,例如油酸、亚油酸、棕榈酸、β-石竹烯、松柏烯硬脂酸、肉豆蔻酸、α-亚麻酸。
根据一个具体实施例,所述活性成分或其组合物包括矿物质,例如钾、钙、镁、磷、铝、铁、钠、硼、锌、镉、硒。
根据一个具体实施例,所述活性成分或其组合物包括维生素,例如硫胺素B1、核黄素B2、吡哆醇B6、氰钴胺素B12、烟酰胺、生物素和抗坏血酸。
根据一个具体实施例,进行甲醇或乙醇提取,例如乙醇浓度为80%;提取时间为1小时;提取温度为40℃;粒径是1.0mm;溶剂与漆树的比例为15:1ml/g。其他提取过程包括但不限于Sakhr and Khatib Heliyon.2020Jan;6(1):e03207中描述的那些过程,其通过引用整体并入本发明。
根据另一实施例,所述植物部分是叶。
本发明还考虑了漆树属植物。
例子包括但不限于:
亚洲和南欧
Rhus chinensis Mill.–中国漆树
Rhus coriaria–坦纳的漆树
Rhus delavayi川芎
澳大利亚,太平洋
Rhus taitensis Guill.(澳大利亚东北部、马里西亚、密克罗尼西亚、法属波利尼西亚)
Rhus sandwicensis A.Gray–内洛(夏威夷)
北美
Rhus aromatica–芳香漆树
Rhus copallinum–有翼或闪亮的漆树
Rhus glabra–光滑的漆树
Rhus integrifolia–柠檬水漆树
Rhus kearneyi–科尔尼漆树
Rhus lanceolata–草原漆树
Rhus michauxii–米修的漆树
Rhus microphylla–沙漠漆树,小叶漆树
Rhus ovata–糖漆树
Rhus trilobata Nutt.–三裂漆树
Rhus typhina–鹿角漆树,伊普里西安,华盛顿
Rhus virens Lindh.ex A.Gray–常绿漆树
人参的化学组成
使用各种方法表征和鉴定人参的化合物,鉴定出人参中的大量化合物,并通常将它们分类为:
·皂苷甙(例如人参皂苷)
·植物甾醇(例如豆甾醇、β-甾醇)
·倍半萜(例如β-阿拉姆烯(β-alamene)和β-芹子烯)
·类黄酮(例如山奈酚)
·聚乙炔(例如人参酚,人参炔A)
·生物碱(例如原阿片碱,吉利宁(girinimbin))
·多糖
·酚类化合物(例如榄香素,蝙蝠葛碱(dauricin),麦芽酚)。
根据具体实施例,人参中的皂苷化合物和多糖化合物是构成其植物化学活性的化合物。发现人参根中最丰富的皂苷化合物是人参皂苷。来自于人参的多糖已被鉴定为NGP、WGP、1-KGP、4-KGP、WGPE和EGP,其中WGP和WGPE是最丰富的,这取决于用于提取的人参植物材料的种类。
大多数人参皂苷属于具有四个反式-环刚性甾体骨架的类固醇家族。它们也被称为人参皂苷,三萜皂苷或达玛烷衍生物。已经从人参植物中分离出超过200个皂苷。除了人参根之外,人参叶和茎、花蕾、果实、浆果和种子中鉴定出了皂苷。因为蒸煮或加热会改变人参产品的皂苷特性,所以在经处理的根、叶、花蕾和浆果中也鉴定出人参皂苷。
人参皂苷被分成几组。两个主要组是糖基连接到C-3和/或C-20的原人参二醇(PPD)-型皂苷和糖基连接到C-6和/或C-20的原人参三醇(PPT)组。其他组包括在C-20具有五元环氧环的奥克梯隆-型、具有非甾体结构的齐墩果烷-型和具有修饰的C-20侧链的达玛烷类型。随着技术被开发用于化学纯化和结构鉴定,新的人参皂苷继续被发现。
下表显示了从不同提取过程制备的人参提取物中回收的人参皂苷化合物:
人参皂苷
a缩写:Hex:正己烷;BuOH:丁醇;CH2Cl2:二氯甲烷;MeOH:甲醇;NH4OAc:乙酸铵;iPrOH:异丙醇;CHCl3:氯仿;EtOAc:乙酸乙酯。
b缩写:TLC:薄层色谱;ELSD:蒸发光散射检测;UV:紫外线。
c缩写:RP:反相;MPLC:中压液相色谱。
下表显示了从人参植物不同部分分离的123种达玛烷-型皂苷的化学式。它们按结构类型的顺序排列。
达马烷型皂苷人参皂苷(Dammarane–type saponin ginsenosides)
人参根(日本人参)的分析表明(每100克根)总脂肪0.17克(0.17%)、钠50毫克、总碳水化合物8.82克(8.82%)包括2.3克膳食纤维和3.85克糖和0.71克(0.71%)蛋白质含量。热量计算表明,100克人参根含有37 kcal。
根据一个具体实施例,所述活性成分或其组合物包括人参皂苷,例如糖基连接到C-3和/或C-20的原人参二醇(PPD)-型皂苷和糖基连接到C-6和/或C-20的原人参三醇(PPT)皂苷,在C-20具有五元环氧环的奥克梯隆-型皂苷、具有非甾体结构的齐墩果烷-型皂苷和达玛烷型皂苷。一些具体的人参皂苷包括但不限于三七皂苷、竹节参皂苷(yesanchinosides)、人参二酮、花人参皂苷(floralginsenosides)和人参皂苷Rg1、Rd、Re、Rb1、R1、Rg3、Rk1、Rf、Rg5、F4、Ro。
根据一个具体实施例,所述活性成分或其组合物包括挥发性化合物,例如萜烃、单萜烃和倍半萜烃,特别是β-阿拉姆烯(β-alamene)和β-硒(β-selenine)。
根据一个具体实施例,所述活性成分或其组合物包括植物甾醇,例如豆甾醇、β-甾醇。
根据一个具体实施例,所述活性成分或其组合物包括聚乙炔,例如人参酚、人参炔A。
根据一个具体实施例,所述活性成分或其组合物包括类黄酮,例如山奈酚。
根据一个具体实施例,所述活性成分或其组合物包括生物碱,例如富马碱、吉林霉素。
根据一个具体实施例,所述活性成分或其组合物包括多糖,例如WGP、KGP-1、KGP-4、WGPE、NGP、EGP。
根据一个具体的实施例,所述活性成分或其组合物包括酚类化合物,例如,榄香素、蝙蝠葛碱、麦芽酚。
根据一个具体实施例,所述活性成分或其组合物包括矿物质,例如钾、钙、镁、磷、铝、铁、钠、硼、锌、镉、硒。
根据一个具体实施例,所述活性成分或其组合物包括维生素,例如维生素D、维生素A和维生素C。
根据一个具体实施例,进行甲醇或乙醇提取,例如乙醇浓度为80%;提取时间为24小时;萃取温度为80-90℃;粒径为1.0mm;溶剂与人参的比例为20:1ml/g。其他提取程序包括但不限于Dong et al.2017Phytother Res Aug;19(8):684-688所述的那些程序,其全部内容通过引用并入本发明。
根据另一个实施例,所述植物部分是叶。
本发明还考虑了人参属植物。
实施例包括但不限于:
适合与本发明一起使用的高丽参栽培品种包括但不限于:Chunpoong、Yunpoong、Gopoong、Sunpoong、Gumpoong、Cheongsun、Sunhyang、Sunun、Sunone、K-1、G-1和Kowon。适合与本发明一起使用的人参栽培品种包括但不限于吉林黄果人参(Jilin HuangguoReshen)、吉参01(Jishen 01)、复兴01(Fuxing 01)、复兴02(Fuxing 02)、康美01(Kangmei01)、新开河01(Xinkaihe 01)、新开河02(Xinkaihe 02)、中农黄凤参(ZhongnongHuangfengshen)和中大宁夏参(Zhongda Linxiashen)。
乳香属(乳香、乳香)的化学成分
乳香,也称为乳香,是一种天然油胶树脂,从乳香树的树皮中渗出。乳香属大约有23种树木,主要生长在阿拉伯、非洲东海岸和印度。使用多种方法表征和鉴定乳香化合物,鉴定出乳香树种的树胶树脂中的大量化合物,并将其分类为:
醇溶性树脂(例如二萜、三萜)
高度芳香的精油(例如单萜和倍半萜)
水溶性胶
根据具体实施例,乳香包含65-85%的醇溶性树脂、约5-9%的高度芳香精油和剩余的水溶性树胶。
在印度,齿叶乳香的主要商业来源是Andhra Pradesh、Gujarat、Madhya Pradesh、Jharkhand和Chhattisgarh。在地区上,它也有不同的名称。齿叶乳香的植物来源和当地名称如下表1所示。Salai,是一种油性胶-树脂,是Boswellia属(家族:Burseraceae)的植物分泌物。它从树干上的切口中取出,然后存放在特制的竹篮中。半固态树胶树脂被允许在篮子中保留大约一个月,在此期间,它的液体在当地被称为“ras”,不断流出。残留物,半固体到固体部分,是一种树胶-树脂,它会慢慢硬化成带有芳香气味的无定形的泪状产品。然后,用木槌或切碎器将其破碎成小块,在此过程中,包括树皮碎片等所有杂质都被手动去除。然后根据其风味、颜色、形状和大小对树胶-树脂进行分级。市面上一般有超细、等级I、等级II和等级III四个等级。从树上获得的新鲜口香糖是热的,味道宜人,味道微苦。它曾是古埃及人、希腊人和罗马人的“乳香”,他们将其用作珍贵的熏香、熏蒸剂和多用途芳香剂。它通常用于制作香粉和香棒。
表1齿叶乳香的植物来源和地方名称
油胶-树脂含有30-60%的树脂,5-10%的精油,可溶于有机溶剂,其余由可溶于水的多糖(~65%阿拉伯糖、半乳糖、木糖)组成。由于精油的存在,树脂具有芬芳的香气,这说明了它们的商业重要性。
根据具体实施例,乳香的共同成分属于萜烯和倍半萜家族,或其萜类衍生物包括但不限于α-和β-蒎烯、α-柠檬烯、月桂烯、芳樟醇、α-荜澄茄油烯、γ-杜松烯、β-波旁烯和乳香中的α-水芹烯二聚体化合物,是构成其植物化学活性的化合物。还鉴定了几种含氧类异戊二烯衍生物,例如羰基衍生物(例如香芹酮、小茴香酮)和含醇萜烯和倍半萜烯衍生物(例如反式松香芹醇、顺式-马鞭草醇和西松烯醇),以及含酯化合物(例如,α-乙酸松油酯和乙酸冰片酯)。
不同的研究人员报告说,柠檬烯是乳香中最丰富的挥发物,而其他人则根据用于提取的乳香树脂的种类鉴定出乙酸辛醇、α-蒎烯和α-侧柏烯是最丰富的。
已从Boswellia ssp中分离出300多种精油。
下表显示了从不同的乳香属植物通过不同提取程序制备的乳香提取物中回收的精油:
虽然许多乳香属植物生产乳香,但商业乳香的主要来源是锯缘乳香(印度)、萨克拉乳香(阿曼)和卡特里乳香(索马里)。
下表显示了按百分比表示的来自不同乳香属植物的乳香的主要成分:
乳脂的一个示例性分析表明了以下成分:
·酸性树脂(6%),溶于酒精,分子式为C20H32O4
·胶(类似于阿拉伯胶)30-36%
·3-乙酰-β-乳香酸(Boswellia sacra)
·α-乳香酸(Boswellia sacra)
·因香酚乙酸酯,C21H34O3
·水芹烯
B.serrata树脂的另一项分析表明,Boswellia serrata的树脂部分含有单萜(α-侧柏烯);二萜类(大环二萜类化合物,例如因香酚、因香酚氧化物、异因香酚氧化物、二萜醇[4,8,12-三甲基-1-异丙基-3,7,11-环十四三烯-1-醇(serratol)]);三萜类(如α-和β-香树脂醇);五环三萜酸(乳香酸);四环三萜酸(甘遂-8,24-二烯-21-酸(tirucall-8,24-dien-21-oic acids))。下表给出了四种主要五环三萜酸(乳香酸)的结构以及四种五环三萜酸(乳香酸)的一些特征:
乳香胶成分含有多糖和聚合物成分。乳香中的蛋白聚糖主要包含主链上的D-半乳糖单元和侧链上的葡萄糖醛酸、糖醛酸、4-O-甲基-葡萄糖醛酸和阿拉伯糖。
根据一个具体实施例,所述活性成分或其组合物包括醇溶性酸性树脂、水溶性树胶、α-乳香酸、因香酚乙酸酯和水芹烯。
根据一个具体实施例,所述活性成分或其组合物包括挥发性化合物,例如α-侧柏烯、杜瓦三烯醇氧化乙酸(Duva-3,9,13-triene-1a-ol-5,8-oxide-1-acetate)、E-β-罗勒烯、醋酸辛醇、醋酸辛酯、柠檬烯、α-蒎烯、辛醇、反式-马鞭草烯醇和松油烯-4-醇。
根据一个具体实施例,所述活性成分或其组合包括矿物质,例如钾、钙、镁、磷、铝、铁、钠、硼、锌、镉、硒。
根据一个具体实施例,进行水或醇提取。
在一些实施例中,所述乳香是通过水提取制备的。本发明描述了示例性水提取:
用水提取制备乳香。起初,乳香被小心地磨成粉。将粉末(25g)与200ml去离子水混合并在室温下以800rpm搅拌过夜。将该混合物以1,500rpm离心10分钟并收集上清液。此后,将上清液再次以2,500rpm离心10分钟,并依次以10,000rpm离心20分钟,然后过滤。滤液可在-20℃储存,然后在-58℃和0.5托冷冻干燥24小时,得到4.02克水溶性提取物。下一步,将所得粉末溶解在100ml甲醇中并在室温下搅拌12小时,然后静置。收集沉淀相并在烘箱中干燥。再次将粉末溶解在去离子水中,反复离心并重新过滤。滤液可以储存,然后冷冻干燥。
在一些实施例中,乳香是通过醇提取制备的。本发明描述了示例性的醇提取:
用酒精提取制备乳香:在该方法中,将100克乳香粉与400毫升甲醇混合。然后将该混合物以650rpm搅拌24小时。所得混合物由两相组成,上层相为酒精且是黄色的,并含有可溶于酒精的物质。然后将该材料在烘箱中在50℃下干燥。底层相是白色沉淀,将其置于烘箱中直至干燥。所得粉末在水中充分溶解,将所得溶液以1,500rpm离心10分钟,收集上清液。此后,将上清液再次以2,500rpm离心10分钟,并依次以10,000rpm离心20分钟,然后过滤。滤液可储存于-20℃,然后冷冻干燥。
其他提取过程包括但不限于Mertens et al,et al.2009,Flavor andFragrance,24:279-30和Hamm et al,Phytochemistry 2005,66:1499-1514中所描述的那些过程,其通过引用整体并入本发明。
本发明还考虑乳香属和其他来自乳香属树的组合物。
实施例包括但不限于:
一些乳香属种类 |
B.socotrana |
B.elongata |
B.ameero |
B.carteri |
B.neglecta |
B.sacra |
B.thurifera |
B.frereana |
B.dioscorides |
B.rivae |
B.papyrifera |
B.serrata |
绞股蓝化学成分
绞股蓝是葫芦科多年生草本植物,叶5裂,果实形似葫芦,不可食用,生长于东北亚、东南亚等许多地区的森林、灌丛或山坡路边,包括中国、韩国、日本、泰国、越南和老挝。绞股蓝也生长在孟加拉国、不丹、印度、印度尼西亚、马来西亚、缅甸、尼泊尔、新几内亚和斯里兰卡。绞股蓝素有“长寿植物”的美誉。使用多种方法表征和鉴定绞股蓝化合物,鉴定出绞股蓝中的多种化合物,并将它们分类为:
·皂苷(例如绞股蓝皂苷)
·酚类化合物
·类黄酮(例如山柰酚、槲皮素、芦丁、欧宾苷、异鼠李素)
·多糖
·甾醇(例如麦角甾烷、胆甾烷、豆甾烷)
·微量元素(例如铜、铁、锌、锰、钴、镍、硒、钼和锶)
·类胡萝卜素
·挥发物(例如丙二酸、苄基-O-β-D-吡喃葡萄糖苷、叶黄素、伏米叶醇、棕榈酸、亚油酸)
根据具体实施例,所述绞股蓝中的皂苷类化合物和多糖类化合物是构成其植物化学活性的化合物。绞股蓝中含量最多的皂苷化合物是绞股蓝皂苷。
大多数绞股蓝皂苷属于三萜皂苷家族。它们也被称为绞股蓝皂苷和达玛烷衍生物。已从绞股蓝植物中分离出150多种皂苷。在绞股蓝的叶和茎、花蕾、果实、浆果和种子中已鉴定出皂苷。
下表显示了来自不同来源的5种不同绞股蓝样品的植物化学特性:
GP1-5代表来自不同来源的绞股蓝样品。数据以每克干植物为基础,以平均值(SD表示。不同字母代表显著差异(P<0.05)。nd代表不可检测。TPC、TSC和TFC代表分别通过光谱法测得的总酚含量、总皂苷含量和总黄酮含量。GAE、GE、RE和QE代表没食子酸当量、绞股蓝当量、芦丁当量和槲皮素当量。芦丁和槲皮素含量是通过HPLC得到黄酮含量。R+Q代表芦丁和槲皮素的总量。
乙醇提取:12g样品溶于250ml 100%乙醇中,在索氏装置中放置5小时。
50%丙酮萃取和75%乙醇萃取:2g样品在环境温度下溶于20ml溶剂中,并通过45微米过滤器过滤。
绞股蓝样品的含水量为3.79至7.57g/100g样品。膳食纤维含量范围为0.6g/g至0.24g/g样品。硒含量在1.7mg/kg到0.94mg/kg之间。
根据一个具体实施例,所述活性成分或其组合物包括绞股蓝皂苷。一些特定的人参皂苷包括但不限于CP-1-6。
根据一个具体实施例,所述活性成分或其组合物包括挥发性化合物,例如丙二酸、苄基-O-β-D-吡喃葡萄糖苷、叶黄素、伏米叶醇、棕榈酸、亚油酸。
根据一个具体实施例,所述活性成分或其组合包括植物甾醇,例如豆甾醇、麦角甾烷。
根据一个具体实施例,所述活性成分或其组合物包括类黄酮,例如山奈酚、槲皮素、芦丁。
根据一个具体实施例,所述活性成分或其组合物包括酚类化合物。
根据一个具体实施例,所述活性成分或其组合包括矿物质,例如钾、钙、镁、磷、铝、铁、钠、硼、锌、镉、硒。
根据一个具体实施例,所述活性成分或其组合包括维生素,例如维生素D、维生素A和维生素C。
根据一个具体实施例,进行甲醇或乙醇提取,例如乙醇浓度为100%或75%;在索氏装置中放置5小时,或50%丙酮提取和75%乙醇提取:在环境温度下将2g样品溶于20ml溶剂中,并通过45微米过滤器过滤。其他提取过程包括但不限于Yantao et al.2016 Chi Med11:43中描述的那些过程,其通过引用整体并入本发明。
根据另一个实施例,所述植物部分是叶。
本发明还考虑的是绞股蓝属植物。
牛至(Origanum Syriacum)
根据一个具体实施例,所述该物种的植物包括黄酮类、类单萜(monoterpenoid)和单萜烯(monoterpene)。已鉴定出超过60种不同的化合物,主要是香芹酚和百里酚,含量超过80%,而含量较少的化合物包括对伞花烃、γ-松油烯、石竹烯、桉油烯醇(spathulenol)、大根香叶烯-D、β-葑醇和δ-松油醇。
下表显示了通过分馏在牛至提取物中鉴定出的有机化合物的概况:
在分析的级分(fraction)中发现的有机化合物的概况。
牛至精油(Ooil)通过蒸汽夹带法获得,油级分通过分馏系统获得。第一个级分在82℃的温度下开始蒸馏,最后一个级分在140℃的温度下蒸馏,最后得到未蒸馏油(Unoil)。在该过程结束时,获得了五个级分,分别命名为级分1(F1)、级分2(F2)、级分3(F3)、级分4(F4)和未蒸馏油(Unoil)。
在HPLC上分析牛至提取物时,鉴定出多种酚类化合物
用HPLC方法测定O.vulgare ssp.vulgare提取物中的酚类化合物。
化合物 | <![CDATA[[M-H]<sup>-</sup>,m/z]]> | <![CDATA[保留时间(t<sub>R</sub>),min]]> | 紫外检测 | 质谱检测 | 浓度(mg/g) |
龙胆酸 | 153 | 3.69±0.04 | NO | YES | <0.02 |
氯原酸 | 353 | 6.43±0.05 | YES | YES | 2.10±0.14 |
对香豆酸 | 163 | 9.48±0.08 | NO | YES | <0.02 |
金丝桃苷 | 463 | 18.60±0.12 | YES | YES | 1.05±0.03 |
异槲皮苷 | 463 | 20.29±0.10 | YES | YES | 0.71±0.19 |
卢丁 | 609 | 20.76±0.15 | YES | YES | 0.64±0.15 |
迷迭香酸 | 360 | 21.80±0.10 | YES | YES | 12.83±2.19 |
槲皮苷 | 447 | 23.64±0.13 | YES | YES | 0.50±0.08 |
栎皮黄素 | 301 | 27.55±0.15 | NO | YES | <0.02 |
藤黄菌素 | 285 | 29.64±0.19 | YES | YES | 0.10±0.04 |
值是平均值±SD(n=3)。
O.vulgare ssp.vulgare提取物中的总多酚含量和抗氧化活性。
每个值是三个独立测量值的平均值±SD。TPC,总多酚含量;SO,超氧化物;GAE,没食子酸当量;RE,芦丁当量;CAE,咖啡酸当量;TE,抗氧化物当量。
本发明还考虑了牛至属植物。
牛至是唇形科多年生草本植物和亚灌木属,原产于欧洲、北非和亚洲温带大部分地区,在开阔或山区生境中发现。少数物种也在北美和其他地区的分散地点归化。
这些植物有强烈的芳香叶子和丰富的管状花,带有持久的彩色苞片。该属包括重要的烹饪草药组:马郁兰(Origanum majorana)和牛至(Origanum vulgare)。
例子包括但不限于:
Origanum acutidens(Hand.-Mazz.)Ietsw.-土耳其、伊拉克
Origanum×adanense Baser&H.Duman-土耳其(O.bargyli×O.laevigatum)
Origanum×adonidis Mouterde-黎巴嫩(O.libanoticum×O.syriacumsubsp.bevanii)
Origanum akhdarense Ietsw.&Boulos-利比亚东部昔兰尼加地区
Origanum amanum Post-土耳其哈塔伊地区
Origanum×barbarae Bornm.-黎巴嫩(O.ehrenbergii×O.syriacumsubsp.bevanii)
Origanum bargyli Mouterde-土耳其、叙利亚
Origanum bilgeri P.H.Davis-土耳其安塔利亚地区
Origanum boissieri Ietsw.-土耳其
Origanum calcaratum Juss.-希腊
Origanum compactum Benth.-西班牙、摩洛哥
Origanum cordifolium(Montbret&Aucher ex Benth.)Vogel-塞浦路斯
Origanum cyrenaicum Bég.&Vacc.-利比亚东部昔兰尼加地区
Origanum dayi Post-以色列
Origanum dictamnus L.–忽布奥勒冈、克里特牛至(Cretan dittany)、克里特岛的白藓(dittany of Crete)-克里特岛特有
Origanum×dolichosiphon P.H.Davis耳其塞汉地区(O.amanum×O.laevigatum)
Origanum ehrenbergii Boiss.-黎巴嫩
Origanum elongatum(Bonnet)Emb.&Maire-摩洛哥
Origanum floribundum Munby-阿尔及利亚
Origanum×haradjanii Rech.f-土耳其(O.laevigatum×O.syriacumsubsp.bevanii)
Origanum haussknechtii Boiss.-土耳其
Origanum husnucan-baseri H.Duman,Aytac&A.Duran-土耳其
Origanum hypericifolium O.Schwarz&P.H.Davis-土耳其
Origanum×intercedens Rech.f.-希腊、土耳其(O.onites×O.vulgaresubsp.hirtum)
Origanum×intermedium P.H.Davis-土耳其代尼兹利地区(O.onites×O.sipyleum)
Origanum isthmicum Danin-西奈半岛
Origanumjordanicum Danin&Kunne-约旦
Origanum laevigatum Boiss.-土耳其、叙利亚、塞浦路斯
Origanum leptocladum Boiss.-土耳其
Origanum libanoticum Boiss.-黎巴嫩
Origanum majorana L.–(甜)马郁兰-土耳其、塞浦路斯;在欧洲、北非、北美和南美的分散地点归化
Origanum×lirium Heldr.ex Halácsy-希腊(O.scabrum×O.vulgaresubsp.hirtum)
Origanum×majoricum Cambess.–耐寒甜马郁兰-西班牙,包括巴利阿里群岛(O.majorana×O.vulgare subsp.virens)
Origanum microphyllum(Benth.)Vogel-克里特岛
Origanum×minoanum P.H.Davis-克里特岛(O.microphyllum×O.vulgaresubsp.hirtum)
Origanum minutiflorum O.Schwarz&P.H.Davis-土耳其
Origanum munzurense Kit Tan&Sorger-土耳其
Origanum×nebrodense Tineo ex Lojac-西西里(O.majorana×O.vulgaresubsp.viridulum)
Origanum onites L.-希腊、土耳其、西西里岛
Origanum×pabotii Mouterde-叙利亚(O.bargyli×O.syriacumsubsp.bevanii)
Origanum pampaninii(Brullo&Furnari)Ietsw-利比亚东部昔兰尼加地区
Origanum petraeum Danin-约旦
Origanum punonense Danin-约旦
Origanum ramonense Danin-以色列
Origanum rotundifolium Boiss.-土耳其、高加索地区
Origanum saccatum P.H.Davis-土耳其
Origanum scabrum Boiss.&Heldr.in P.E.Boissier-希腊
Origanum sipyleum L.-土耳其、希腊群岛
Origanum solymicum P.H.Davis-土耳其安塔利亚地区
Origanum syriacum L.-土耳其、塞浦路斯、叙利亚、黎巴嫩、约旦、巴勒斯坦、以色列、西奈半岛、沙特阿拉伯
Origanum vetteri Briq.&Barbey-克里特岛
Origanum vogelii Greuter&Burdet-土耳其
Origanum vulgare L.-牛至-欧洲、北非、温带亚洲(伊朗、西伯利亚、中亚、中国等);在北美、新西兰、委内瑞拉的部分地区归化。
根据一个具体实施例,所述活性成分或其组合物包括牛至提取物的有机化合物成分。
根据一个具体的实施例,所述活性成分或其组合物选自α-侧柏烯、α-蒎烯、β-月桂烯、水芹烯、α-松油烯、邻伞花烃、柠檬烯、1,8-桉树脑、γ-松油烯、百里酚、香芹酚、反式石竹烯和α-蛇麻烯。
根据一个具体实施例,所述活性成分或其组合物包括单萜烃、氧化单萜和倍半萜烃。
根据一个具体实施例,所述活性成分或其组合物包括酚类化合物,例如龙胆酸、绿原酸、对香豆酸、金丝桃苷、异槲皮苷、芦丁、迷迭香酸、槲皮素、槲皮素和木犀草素。
根据一个具体实施例,所述活性成分或其组合包括矿物质,例如钾、钙、镁、磷、铝、铁、钠、硼、锌、镉、硒。
芝麻
芝麻含有木脂素、芝麻素、芝麻素、松脂醇和落叶松树脂醇。不溶性11S球蛋白和可溶性2S白蛋白,通常称为α-球蛋白和β-球蛋白,是两种主要的储存蛋白,占芝麻种子总蛋白的80-90%。氨基酸组成的比较表明,它们的疏水性明显低于已知的油质蛋白,因此不应是油质蛋白的聚集多聚体。对芝麻蛋白的免疫识别结果表明,这三种多肽是成熟种子细胞器活性组装过程中伴随着油质蛋白和三酰基甘油而聚集在油体中的独特蛋白质。已发现磷脂、油酸和亚油酸、叶绿素和芝麻素、芝麻酚和γ-生育酚。10种化合物[2-糠基硫醇、2-苯乙基硫醇、2-甲氧基苯酚、4-羟基2,5-二甲基-3[2H]-呋喃酮、2-戊基吡啶、2-乙基-3,5-二甲基吡嗪、乙酰吡嗪、[E,E]-2,4-癸二烯醛、2-乙酰基-1-吡咯啉和4-乙烯基-2-甲氧基苯酚]被量化。基于油中的高OAV,特别是2-乙酰基-1-吡咯啉[烘焙]、2-糠硫醇[咖啡样]、2-苯乙基硫醇[橡胶]和4-羟基-2,5-二甲基3[2H]-呋喃酮[类似焦糖的]被阐明是造成碎芝麻材料的整体烤的、硫磺的气味的重要因素。从芝麻籽中分离得到的新型芝麻素糖苷的结构确定为芝麻素2'-O-β-d-吡喃葡萄糖苷、芝麻素2'-O-β-d-吡喃葡萄糖[1→2]-O-β-d-吡喃葡萄糖苷和芝麻素2'-O-β-d-吡喃葡萄糖基[1>>2]-O-[β-d-吡喃葡萄糖基[1>>6]]-[β-吡喃葡萄糖苷。还有少量芝麻木脂素,例如-(7S,8'R,8R)-茴香内酯(acuminatolide)胡椒醇和松脂醇(如前所述)。
本发明还考虑芝麻属植物。
例子包括但不限于:
Sesamum abbreviatum Merxm.
Sesamum alatum Thonn.
Sesamum angolense Welw.
Sesamum biapiculatum De Wild.
Sesamum calycinum Welw.
Sesamum capense Burm.f.
Sesamum digitaloides Welw.ex Schinz
Sesamum gracile Endl.
Sesamum hopkinsii Suess.
Sesamum indicum L.
Sesamum lamiifolium Engl.
Sesamum latifolium J.B.Gillett
Sesamum lepidotum Schinz
Sesamum macranthum Oliv.
Sesamum marlothii Engl.
Sesamum mombazense De Wild.&T.Durand
Sesamumparviflorum Seidenst.
Sesamumpedalioides Welw.ex Hiern
Sesamum radiatum Schumach.&Thonn.
Sesamum rigidum Peyr.
Sesamum rostratum Hochst.
Sesamum sabulosum A.Chev.
Sesamum schinzianum Asch.
Sesamum somalense Chiov.
Sesamum thonneri De Wild.&T.Durand
Sesamum triphyllum Welw.ex Asch.
根据本发明的一些实施例,所述含有木脂素的植物包括多种植物性食物,包括种子(亚麻、南瓜、向日葵、芝麻)、全谷物(黑麦、燕麦、大麦)、麸皮(小麦、燕麦、黑麦)、豆类、水果(尤其是浆果)和蔬菜(西兰花和羽衣甘蓝是木脂素的丰富来源。其他蔬菜,如白甘蓝和红甘蓝、球芽甘蓝、花椰菜、胡萝卜、绿色和红色甜椒也是很好的来源)。
其他含有芝麻素的植物包括但不限于刺五加。
因此,考虑上述植物的任何组合物,包括2、3、4、5、6、7种植物。根据另一个实施例,提取物或级分的组合物包括2、3、4、5、6、7种不同植物。
示例包括但不限于黑种草Nigella sativa、银斑百里香Thymus vulgaris、牛至Origanum syriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamumindicum和乳香Rhus coriaria。
黑种草Nigella sativa、头状百里香Thymus capitatus、牛至Origanumsyriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamum indicum乳香Rhus coriaria。
黑种草Nigella sativa、头状百里香Thymus capitatus、银斑百里香Thymusvulgaris、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamum indicum和and乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Satujera thymbra,芝麻Sesamum indicum和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Thymbra spicata,芝麻Sesamum indicum和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Thymbra spicata,百里香Satujera thymbra,和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Thymbra spicata,百里香Satujera thymbra,芝麻Sesamum indicum。
黑种草Nigella sativa,牛至Origanum syriacum,百里香Thymbra spicata,百里香Satujera thymbra,芝麻Sesamum indicum和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,百里香Satujera thymbra,芝麻Sesamum indicum和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,芝麻Sesamum indicum和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Thymbra spicata,和乳香Rhus coriaria。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris,牛至Origanum syriacum,百里香Thymbra spicata,百里香Satujera thymbra。
黑种草Nigella sativa,头状百里香Thymus capitatus。
黑种草Nigella sativa,银斑百里香Thymus vulgaris。
黑种草Nigella sativa,牛至Origanum syriacum。
黑种草Nigella sativa,百里香Thymbra spicata。
黑种草Nigella sativa,百里香Satujera thymbra。
黑种草Nigella sativa,芝麻Sesamum indicum。
黑种草Nigella sativa,乳香Rhus coriaria。
本发明还考虑了没有黑种草的各种组合物。
根据另一个实施例,所述活性成分的组合物,例如百里醌、香芹酚、百里酚;百里醌,香芹酚;百里醌,百里酚;香芹酚、百里酚。
黑种草Nigella sativa,头状百里香Thymus capitatus,银斑百里香Thymusvulgaris。
黑种草Nigella sativa,银斑百里香Thymus vulgaris,牛至Origanum syriacum。
黑种草Nigella sativa,牛至Origanum syriacum,百里香Thymbra spicata。
黑种草Nigella sativa,百里香Thymbra spicata,百里香Satujera thymbra。
黑种草Nigella sativa,百里香Satujera thymbra,芝麻Sesamum indicum,乳香Rhus coriaria。
根据一些实施例,所述植物及其活性成分列于下表中。
本发明考虑其它实施例,其包含Nigella sativa、Thymus capitatus、Thymusvulgaris、Origanum syriacum、Thymbra spicata、Satujera thymbra、Sesamum indicum、Rhus coriaria、Panaxginseng和Gynostemmepentaphyllum植物的任何一种或其属中2、3、4、5、6、7和8种植物的组合物。
本发明的方法、疫苗、药物组合物、组合物或食品补充剂的其他实施例进一步包含大麻或大麻素。
根据本发明的一个方面,提供了一种食品补充剂、组合物或提取物,进一步包括“贝都因茶”,其中,“贝都因茶”包括玫瑰叶姜味草灌木(Rose Leaves Micromeriafruticose),鼠尾草(Salvia),柠檬草(cymbopgon)(柠檬醛)阿莱藤属(Aloysia),马鞭草(verbena officinalis),马郁兰(origanum majorana),薄荷。
根据本发明的一个方面,提供了一种食品补充剂、组合物或提取物,进一步包括“贝都因茶”,其中,“贝都因茶”包括百里香、鼠尾草、小豆蔻、肉桂、红茶、哈布克、马尔马亚。
植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物可用于实体瘤和软瘤以及增殖性疾病的治疗。
如本发明所用,术语“实体瘤和软瘤以及增殖性疾病”是指包含囊肿或液体的细胞/组织的异常生长。实体瘤和软瘤以及增殖性疾病可能是良性的(而非癌性的)或恶性的(癌性的)。不同类型的实体瘤和软瘤以及增殖性疾病以形成它们的细胞类型命名。实体瘤和软瘤以及增殖性疾病的例子有肉瘤、癌和淋巴瘤。“肉瘤”是发生于结缔组织或支撑组织(如骨骼或肌肉)的癌症。“癌”是由排列在身体组织中的腺细胞和上皮细胞引起的癌症。“淋巴瘤”是淋巴结、脾脏和胸腺等淋巴器官的癌症。由于这些细胞出现在身体的大多数组织中,淋巴瘤可能在各种各样的器官中发展。这里所考虑的典型实体瘤和软瘤以及增殖性疾病包括但不限于肉瘤和癌,例如纤维肉瘤、黏液肉瘤、脂肪肉瘤、软骨肉瘤、骨原性肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤、尤文氏瘤、平滑肌肉瘤、横纹肌肉瘤、结肠癌、胰腺癌、乳腺癌、卵巢癌、前列腺癌、鳞状细胞癌、基底细胞癌、腺癌、汗腺癌、皮脂腺癌、乳头状癌、乳头状腺癌、囊腺癌、髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎癌、维尔姆斯氏瘤、宫颈癌、睾丸癌、肺癌、小细胞肺癌、膀胱癌、上皮癌、多形性胶质母细胞瘤、星形细胞瘤、髓母细胞瘤、颅咽管瘤、室管膜瘤、松果体瘤、血管母细胞瘤、听神经瘤、少突胶质母细胞瘤、皮肤T细胞淋巴瘤(CTCL)、黑色素瘤、神经母细胞瘤和视网膜母细胞瘤。
实体瘤和软瘤以及增殖性疾病可在身体的肌肉、骨骼、淋巴系统、骨髓和器官中发展。例如间皮瘤、肉瘤、淋巴瘤、肉瘤以及乳腺癌、前列腺癌、肾癌、卵巢癌、胰腺癌、甲状腺癌和结肠癌。
此外,放射或化疗治疗血癌可导致继发性实体瘤和软瘤以及增殖性疾病。事实上,在癌症幸存者中,实体瘤是继治疗后第二常见的肿瘤类型。
实体瘤和软瘤以及增殖性疾病的分类方式在了解癌症的病理、确定最重要的治疗过程和评估患者的预后方面起着重要的作用。
基于病理学家在肿瘤细胞中发现的异常和肿瘤扩散的可能性的分级用来对实体瘤和软瘤以及增殖性疾病进行分类。与正常、健康细胞和组织的组织结构相似、增殖相对缓慢的肿瘤组织被称为“良好分化”。快速增殖的肿瘤细胞看起来不正常,缺乏正常的组织结构,被称为“未分化”或“低分化”。肿瘤一般分为四种等级:
·病理学家通常将细胞与正常细胞相似且增殖缓慢的肿瘤归类为1级肿瘤。
·2级肿瘤细胞结构异常较多,细胞分化中等,复制速度较1级肿瘤快。
·分类为3级或“高级别”的肿瘤细胞组织分化差,扩散速度快于1级和2级肿瘤。
·4级肿瘤完全缺乏细胞分化,看起来与健康细胞和低级别肿瘤截然不同。
虽然许多癌症使用这种分级系统进行分类,但值得注意的是,一些实体瘤和软瘤以及增殖性疾病类型使用其他分级系统进行定义。
例如,医生可以根据有丝分裂率、乳管中肿瘤活动的程度(小管形成)和肿瘤细胞中细胞核的大小和形状(称为核级别)对乳腺癌进行分类。这三个类别中的每一个都接收范围从1到3不的分数。1分表示肿瘤组织更接近健康细胞和组织。得分为“3”与具有最异常外观的细胞和组织相关。在为这三个类别分别赋值后,将这些值相加,得到一个从3到9的综合分数(composite score)。这些值落入三个不同的肿瘤类型中:
·低等级或分化良好的肿瘤的综合评分为3-5分。
·肿瘤被鉴定为中级或中度分化,评分从6到7。
·得到8或9分的肿瘤被认为是低分化的。
肿瘤学界使用格里森评分系统对前列腺活检样本的病理结果进行前列腺癌分级。病理学家将患病组织的外观与健康组织进行比较,并为该组织分配1到5的分数。在肿瘤中最常见的异常组织被称为主要模式,而次要模式是下一个最常见的组织模式。
将主要和次要模式的分数相加,形成格里森分数(Gleason score),其结果落入四个类别中:
·格里森X表示病理学家无法确定格里森评分。
·格里森2-6与分化良好的肿瘤组织相关。
·格里森7分用于定义中度分化肿瘤组织。
·格里森8-10分表示肿瘤组织分化不良或完全未分化。
在某些实例中,所述实体瘤和软瘤以及增殖性疾病是纤维肉瘤、黏液肉瘤、脂肪肉瘤、软骨肉瘤、骨原性肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤、尤文氏瘤、平滑肌肉瘤、横纹肌肉瘤、结肠癌、胰腺癌、乳腺癌、卵巢癌、前列腺癌、鳞状细胞癌、基底细胞癌、腺癌、汗腺癌、皮脂腺癌、乳头状癌、乳头状腺癌、囊腺癌、髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎癌、维尔姆斯氏瘤、宫颈癌、睾丸癌、肺癌、小细胞肺癌、膀胱癌、上皮癌、多形性胶质母细胞瘤、星形细胞瘤、髓母细胞瘤、颅咽管瘤、室管膜瘤、松果体瘤、血管母细胞瘤、听神经瘤、少突胶质细胞瘤、皮肤T细胞淋巴瘤(CTCL)、黑色素瘤、神经母细胞瘤和视网膜母细胞瘤。
在其他实施例中,所述实体瘤和软瘤以及增殖性疾病是脑癌、乳腺癌、三阴性乳腺癌、膀胱癌、骨癌、结直肠癌、肺癌、肾癌、肝癌、胃癌、前列腺癌、肉瘤、黑色素瘤、癌症或淋巴瘤。
在一些实施例中,所述实体瘤和软瘤以及增殖性疾病是前列腺癌、乳腺癌、结直肠癌、胰腺癌或淋巴瘤。
在一些实施例中,所述实体瘤和软瘤以及增殖性疾病是淋巴瘤。
根据本发明的一些实施例,所述增殖性疾病是纤维瘤。
根据本发明的一些实施例,所述增殖性疾病是子宫内膜异位症。
本发明的一种或多种植物衍生的组分可以与其他药物共同施用以增加治疗生物利用度、提高治疗功效和最小化副作用。本发明的一种或多种植物衍生的组分可以以线性或循环的形式给药,或以任何生理学上被认为适合作为递送治疗手段的构象给药。
联合治疗
在治疗、预防、改善、控制或减少实体瘤和软瘤以及增殖性疾病的生长和转移时,本发明的化合物和/或组分可以与下列物质一起使用:(1)癌症疫苗接种策略,(2)免疫检查点调节剂例如免疫检查点抑制剂的拮抗抗体(抗-PD1,抗-PD-L1,抗-CTLA4,抗-Tim3,抗-VISTA,抗-KIR)或抗免疫加速剂的激动抗体(抗-Lag3,抗-OX40,抗-ICOS,抗-4-1BB),(3)抗在转化细胞中通常上调的细胞表面蛋白的阻断或消耗抗体(CEACAM1,Syndecan-2,GRP78),(4)抗血管生成治疗(抗-VEGF,抗-VEGFR,VEGFR小分子抑制剂),(5)抗淋巴血管生成(抗VEGF、FDF2、PDGF及其各自受体的阻断抗体或抑制剂),(6)标准化疗治疗(例如吉西他滨,紫杉醇,FOLFORINOX),(7)照射治疗,(8)趋化因子拮抗剂(CCR1、CCR4、CCR6、CXCR4、CXCR2、CXCR7小分子抑制剂,阻断抗体,或消耗抗体),(9)抗癌症中常见体细胞突变的抑制剂,例如专门针对以下基因(BRAF、KRAS、NRAS、EGFR、CTNNB1、NOTCH1、PIK3CA、PTEN、APC、FLT3、IDH1、IDH2、KIT、TP53、JAK2)的抑制剂。
在一些实施例中,所述化疗治疗剂选自醋酸阿比特龙、阿法替尼、阿地白介素、阿仑单抗、阿利维A酸、阿曲他明、氨磷汀、氨基谷氨酰胺阿那格雷、阿那曲唑、三氧化二砷、天冬酰胺酶、阿扎胞苷、硫唑嘌呤、苯达莫司汀、贝伐单抗、贝沙罗汀、比卡鲁胺、博来霉素、硼替佐米、白消安、卡培他滨、卡铂、卡莫司汀、西妥昔单抗、苯丁酸氮芥、顺铂、克拉屈滨、克唑替尼、环磷酰胺、阿糖胞苷、达卡巴嗪、放线菌素、达沙替尼、柔红霉素、地尼白霉素、地西他滨、多西他赛、地塞米松、去氧氟尿苷、阿霉素、表柔比星、促红细胞生成素(EpoetinAlpha)、埃博霉素、厄洛替尼、雌莫司汀、恩替诺特(Etinostat)、依托泊苷、依维莫司、依西美坦、非格司亭、氟尿嘧啶、氟达拉滨、氟尿嘧啶、氟甲睾酮、氟他胺、叶酸连接生物碱、吉非替尼、吉西他滨、吉姆单抗奥佐米星、GM-CT-01、戈舍瑞林、六甲基三聚氰胺、羟基脲、伊布单抗、伊达比星、异环磷酰胺、伊马替尼、干扰素α、干扰素β、伊立替康、伊沙匹隆、拉帕替尼、亚叶酸、亮丙瑞林、来那度胺、来曲唑、洛莫司汀、甲氯乙胺、甲地孕酮、美法仑、巯嘌呤、甲氨蝶呤、丝裂霉素、米托蒽醌、奈拉滨、尼罗替尼、尼鲁米特、奥曲肽、奥法木单抗、奥普利维金、奥沙利铂、紫杉醇、帕尼单抗、培美曲塞、喷司他丁、多糖半乳凝素抑制剂、丙卡巴肼、雷洛昔芬、维甲酸、利妥昔单抗、罗米洛汀(Romiplostim)、沙格莫司汀(Sargramostim)、索拉非尼、链脲佐菌素、舒尼替尼、他莫昔芬、替西罗莫司、替莫唑胺、替尼泊苷、沙利度胺、硫鸟嘌呤、塞替派、硫鸟嘌呤、托泊替康、托瑞米芬、托西莫单抗、曲美替尼、曲妥珠单抗、维甲酸、缬柔比星、VEGF抑制剂和抗酒石酸酸性磷酸酶(trap)、长春碱、长春新碱、长春地辛、长春瑞滨、长春花内酯(Vintafolide)(EC145)、伏立诺他,其盐,以及其任何组合物。
在其他实施例中,所述治疗性抗体选自阿巴伏单抗(Abagovomab)、阿拉赛珠单抗-聚乙二醇(Alacizumab-pegol)、阿仑单抗(Alemtuzumab)、铟标喷替酸阿妥莫单抗(Altumomab pentetate)(Hybri-ceaker)、阿玛妥昔单抗(Amatuximab)、马安那莫单抗(Anatumomab mafenatox)、抗-PD-1抗体、阿泊珠单抗(Apolizumab)、阿西莫单抗(Arcitumomab)(CEA-Scan)、贝利单抗(Belimumab)、贝伐单抗(Bevacizumab)、比伐珠单抗美登素(Bivatuzumab mertansine)、博纳吐单抗(Blinatumomab)、本妥昔单抗(Brentuximab-vedotin)、坎妥珠单抗美登素(Cantuzumab mertansine)、坎妥珠单抗美登素(Cantuzumab ravtansine)、卡罗单抗喷地肽(Capromab pendetide)(Prostascint),卡妥索单抗(Catumaxomab)(Removab)、西妥昔单抗(Erbitux)、泊西他组单抗(Citatuzumabbogatox)、西妥木单抗(Cixutumab)、克利妥珠单抗-轮环藤宁四乙酸(Clivatuzumab-tetraxetan)(hPAM4-Cide)、可那木单抗(Conatumumab)、达罗托组单抗(Dalotuzumab)、地舒单抗(Denosumab)、曲齐妥单抗(Drozitumab)、依决洛单抗(Edrecolomab)(Panorex)、依那妥组单抗(Enavatuzumab)、吉妥珠单抗(Gemtuzumab)、替伊莫单抗(Ibritumomabtiuxetan)、伊匹单抗(Ipilimumab)(MDX-101)、奥法木单抗(Ofatumumab)、帕尼单抗(Panitumumab)、利妥昔单抗(Rituximab)、托西莫单抗(Tositumomab)、曲妥珠单抗(Trastuzumab)及其任何组合物。
在进一步的实施例中,所述化疗剂是放射性同位素、胸苷酸合酶抑制剂、铂化合物、长春花生物碱剂或其任何组合物。
在一些实施例中,本发明所述的化合物和/或组分可与抗炎或镇痛剂如阿片激动剂、脂氧合酶抑制剂如5-脂氧合酶抑制剂、环氧合酶抑制物如环氧合酶类-2抑制物、白介素抑制物如白介素-1抑制物、NMDA拮抗剂、一氧化氮抑制剂或一氧化氮合成抑制剂、非甾体抗炎剂或细胞因子抑制抗炎剂,例如与化合物如对乙酰氨基酚、阿司匹林、可待因、生物TNF螯合剂、芬太尼、布洛芬、吲哚美辛、酮咯酸、吗啡、萘普生、非那西丁、吡罗昔康、一种甾体镇痛剂、舒芬太尼、孙林达克、替尼达普等联合使用。
在一些实施例中,所述PD-1和/或PD-L1抑制剂选自度伐鲁单抗(durvalumab)、阿特珠单抗(atezolizumab)、派姆单抗(pembrolizumab)、纳武单抗(nivolumab)、AP-106、AP-105、MSB-2311、CBT-501、阿维单抗(avelumab)、AK-105、IO-102、IO-103、PDR-001、CX-072、SHR-1316、JTX-4014、GNS-1480、重组人源化抗-PD1单抗(上海君实生物医药科技股份有限公司)、REGN-2810、佩拉雷普(Pelarorep)、SHR-1210、PD1/PDL1抑制剂疫苗(THERAVECTYS)、BGB-A317、重组人源化抗-PD-1单克隆抗体(Bio-Thera Solutions)、靶向PD-1的前抗体(Probody)(CytomX)、XmAb-20717、FS-118、PSI-001、SN-PDL01、SN-PD07、PD-1修饰TIL(Sangamo Therapeutics)、PRS-332、FPT-155、杰诺(jienuo)单抗(Genor Biopharma)、TSR-042、REGN-1979、REGN-2810、瑞诺司他(resminostat)、FAZ-053、PD-2/CTLA-4双特异性抗体(MacroGenics)、MGA-012、MGD-013、M-7824、基于PD-1的双特异性抗体(北京韩美药品有限公司药)、AK-112、AK-106、AK-104、AK-103、BI-754091、ENUM-244C8、MCLA-145、MCLA-134、抗-PD1溶瘤单克隆抗体(Transgene SA)、AGEN-2034、IBI-308、WBP-3155、JNJ-63723283、MEDI-0680、SSI-361、CBT-502、抗-PD-1双特异性抗体、双靶向抗-PD-2/LAG-3单克隆抗体(TESARO)、双靶向抗-PD-1/TIM-3单克隆抗体(TESARO)、PF-06801591、LY-3300054、BCD-100、STI-1110、派姆单抗(pembrolizumab)生物仿制药、纳武单抗(nivolumab)生物仿制药、PD-L1-TGF-β治疗、KY-1003、STI-1014、GLS-010、AM-0001、GX-P2、KD-033、PD-L1/BCMA双特异性抗体(Immune Pharmaceuticals)、PD-1/Ox40靶向双特异性的抗体(ImmunePharmaceuticals)、BMS-936559、抗PD-I/VEGF-A DARPins(Molecular Partners)、mDX-400、ALN-PDL、PD-1抑制剂肽(Aurigene)、siRNA负载的树突状细胞疫苗(AlnylamPharmaceutical)、GB-226、PD-L1靶向基于CAR-TNK免疫疗法(PD-L1 targeting CAR-TNK-based immunotherapy)(NK Therapeutics/NantKwest)、INSIX-RA、INDUS-903、AMP-224、抗-CTLA-4/抗-PD-1双特异性人源化抗体(Akeso Biopharma)、B7-H1疫苗(肿瘤生物学国家重点实验室/第四军医大学)和GX Dl。
在一些实施例中,所述PD-1抑制剂是选自纳武单抗、派姆单抗(Pembrolizumab)和匹利珠单抗(Pidilizumab)的抗体。
在一些实施例中,所述免疫检查点抑制剂是CTLA-4抑制剂。本领域已知许多CTLA-4抑制剂。在一些实施例中,所述CTLA-3抑制剂是抗体。在一些实施例中,所述CTLA-4抑制剂抗体选自伊匹单抗、曲美木单抗(Tremelimumab)、AGEN1884和AGEN2041。在一些实施例中,所述CTLA4抑制剂抗体是伊匹单抗。在一些实施例中,所述CTLA-4抑制剂抗体是曲美木单抗。在一些实施例案中,所述CTLA-4抑制剂抗体是AGEN1884。在一些实施例中,所述CTLA4抑制剂抗体是AGEN2041。
术语“治疗”是指抑制、预防或阻止病理(疾病、病症或病状)的发展和/或引起病理的减少、缓解或消退。本领域技术人员将理解可以使用各种方法和测定来评估病理学的发展,并且类似地,可以使用各种方法和测定来评估病理学的减少、缓解或消退。
如本发明所用,术语“预防”是指防止疾病、病症或病状在可能处于疾病风险中但尚未被诊断为患有该疾病的受试者中发生。
如本发明所用,术语“受试者”包括患有该病的任何年龄或性别的哺乳动物,优选人类,男性或女性。优选地,该术语包括有患上该病的风险的个体(例如,65岁以上、暴露于香烟烟雾、致癌物、对实体瘤和软瘤以及增殖性疾病的家族易感性)。
本发明所述物质组合物包含组分(植物物种或其属衍生的组分,所述组分选自包含植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或它们的组合物的组,其中所述组分能够治疗实体瘤和软瘤以及增殖性疾病)可以施用给受试者,或者以药物组合物与合适的载体或赋形剂混合的形式施用给受试者。
如本文所用,“药物组合物”是指本发明所述的一种或多种活性成分与其他化学组分如生理上合适的载体和赋形剂的制剂。药物组合物的目的是促进将化合物施用于生物体。
在本发明中,术语“活性成分”是指包含负责生物效应的组分的物质组合物。
在下文中,短语“生理学上可接受的载体”和“药学上可接受的载体”可以互换使用,是指不会对生物体造成显着刺激并且不会消除所施用化合物的生物活性和性质的载体或稀释剂。所述短语包括佐剂。
本发明中的术语“赋形剂”是指添加到药物组合物中以进一步促进活性成分给药的惰性物质。所述赋形剂的示例但不限于碳酸钙、磷酸钙、各种糖和淀粉类型、纤维素衍生物、明胶、植物油和聚乙二醇。
可在“Remington’s Pharmaceutical Sciences”,Mack Publishing Co.,Easton,PA的最新版本中找到用于药物的配制和给药的技术,其通过引用并入本发明。
合适的给药途径可以例如包括口服、直肠、经粘膜,尤其是经鼻、肠或肠胃外给药,包括肌肉内、皮下和髓内注射以及鞘内、直接心室内、心内给药,例如进入右心室或左心室腔,进入冠状动脉,静脉内,腹腔内,鼻内,或肺内或眼内注射。
在本发明的各种实施例中,组合物作为适合口服给药的药物或膳食补充剂剂型提供。适合口服给药的剂型包括片剂、软胶囊、硬胶囊、丸剂、颗粒剂、散剂、乳剂、混悬剂、喷雾剂、糖浆剂和丹剂。在本发明的各种其他实施例中,组合物作为适合于肠胃外给药的药物剂型提供,例如用于作为滴剂或注射给药的液体制剂,或作为用于栓剂的固体或半固体剂型。
将药物输送到中枢神经系统(CNS)的常规方法包括:神经外科策略(例如,脑内注射或脑室内输注);制剂的分子操作(例如,产生嵌合融合蛋白,包含对内皮细胞表面分子具有亲和力的转运多肽与本身不能穿过血脑屏障的制剂组合)以尝试利用血脑屏障的内源性运输途径中的一种;旨在增加药剂脂溶性的药理学策略(例如,将水溶性药剂与脂质或胆固醇载体结合);以及通过高渗破坏(由将甘露醇溶液注入颈动脉或使用生物活性剂如血管紧张素多肽导致)而暂时破坏血脑屏障的完整性。然而,这些策略中的每一个都具有局限性,例如与侵入性外科手术相关的固有风险、由内源性转运系统中固有限制所施加的尺寸限制、与全身施用包含可能在中枢神经系统之外活跃的载体基序的嵌合分子相关的潜在不良生物学副作用,以及在血脑屏障被破坏的大脑区域内可能存在脑损伤的风险,这使其成为一种次优的递送方法。
可替代地,可以以局部而非全身方式施用药物组合物,例如,通过将药物组合物直接注射到患者的组织区域中。
本发明的一些实施例的药物组合物可以通过本领域公知的方法制造,例如,通过常规的混合、溶解、制粒、糖衣丸制造、研磨、乳化、包封、包埋或冻干方法。
根据本发明的一些实施例使用的药物组合物,因此可以使用生理学上可接受的载体包括赋形剂和助剂的一种或多种以常规方式配制,其有助于将活性成分加工成可药用的制剂。适当的配方取决于所选择的给药途径。
对于注射剂,药物组合物的活性成分可以配制在水溶液中,优选在生理相容的缓冲液中,例如汉克溶液、林格溶液或生理盐缓冲液。对于经粘膜给药,制剂中使用适合待渗透屏障的渗透剂。这种渗透剂在本领域中通常是已知的。
对于口服给药,通过将活性化合物与本领域熟知的药学上可接受的载体结合可便于配制药物组合物。此类载体使药物组合物能够配制成片剂、丸剂、糖衣丸、胶囊、液体、凝胶、糖浆、浆液、悬浮液等,以供患者口服摄入。可使用固体赋形剂制备口服药用制剂,任选研磨所得混合物,并加工颗粒混合物,如果需要,在添加合适的助剂后,以获得片剂或糖衣丸芯。合适的赋形剂尤其是填充剂,例如糖,包括乳糖、蔗糖、甘露糖醇或山梨糖醇;纤维素制剂,例如玉米淀粉、小麦淀粉、大米淀粉、马铃薯淀粉、明胶、黄蓍胶、甲基纤维素、羟丙基甲基纤维素、碳甲基纤维素钠;和/或生理上可接受的聚合物,例如聚乙烯吡咯烷酮(PVP)。如果需要,可以添加崩解剂,例如交联聚乙烯吡咯烷酮、琼脂或海藻酸或其盐例如海藻酸钠。
糖衣丸芯配有合适的涂层。为此目的,可使用浓缩糖溶液,可任选含有阿拉伯胶、滑石粉、聚乙烯吡咯烷酮、卡波姆凝胶、聚乙二醇、二氧化钛、漆溶液和合适的有机溶剂或溶剂混合物。可以将染料或颜料添加到片剂或糖衣丸包衣中用于鉴定或表征活性化合物剂量的不同组合物。
可口服使用的药物组合物包括由明胶制成的推入式胶囊以及由明胶和增塑剂如甘油或山梨糖醇制成的软、密封胶囊。推入式胶囊可包含与填充剂如乳糖、粘合剂如淀粉、润滑剂如滑石粉或硬脂酸镁以及任选的稳定剂混合的活性成分。在软胶囊中,活性成分可以溶解或悬浮在合适的液体中,例如脂肪油、液体石蜡或液体聚乙二醇。此外,可以添加稳定剂。用于口服给药的所有制剂的剂量应适合所选给药途径。
对于口腔给药,组合物可以采用以常规方式配制的片剂或锭剂的形式。
在具体实施例中,本发明的组分和/或组合物以适合通过吸入或鼻腔给药的形式提供。
对于通过鼻吸入给药,根据本发明的一些实施例使用的活性成分通过使用合适的推进剂例如二氯二氟甲烷、三氯氟甲烷、二氯四氟乙烷或二氧化碳以气溶胶喷雾剂的形式从加压包或喷雾器方便地递送。在加压气雾剂的情况下,可以通过提供阀门确定剂量单位以输送计量的量。用于分配器的例如明胶的胶囊和药筒可配制成包含化合物和合适的粉末基质例如乳糖或淀粉的粉末混合物。
本发明所述的药物组合物可配制用于肠胃外给药,例如通过弹丸式注射或持续输注。注射用制剂可以表现为单位剂量形式,例如,在安瓿或多剂量容器中,任选一种添加防腐剂。所述组合物可以是在油性或水性载体中的悬浮液、溶液或乳液,并且可以包含配方剂,例如悬浮剂、稳定剂和/或分散剂等。
用于肠胃外给药的药物组合物包括水溶性形式的活性制剂的水溶液。此外,活性成分的悬浮液可以制备成合适的油性或水基注射悬浮液。合适的亲脂性溶剂或载体包括脂肪油如芝麻油,或合成脂肪酸酯如油酸乙酯、甘油三酯或脂质体。水性注射悬浮液可能含有增加悬浮液粘度的物质,例如羧甲基纤维素钠、山梨糖醇或葡聚糖。任选地,悬浮液还可包含合适的稳定剂或增加活性成分溶解度的化学试剂以允许制备高度浓缩的溶液。
可替代地,活性成分可以是粉末形式,用于在使用前与合适的载体例如,无菌、无热原的水基溶液结合。
本发明的一些实施例的药物组合物也可以配制成直肠组合物例如栓剂或保留灌肠剂例如,常规栓剂基质如可可脂或其他甘油酯。
适用于本发明的一些实施例的上下文中的药物组合物包括含有有效量的以实现预期目的活性成分的组合物。更具体地,治疗有效量是指有效预防、减轻或改善病症(例如实体瘤和软瘤以及增殖性疾病)的症状或进展或延长接受治疗的受试者存活的活性成分(包含负责生物效应的组分的物质组合物)的量。
治疗有效量的确定完全在本领域技术人员的能力范围内,尤其是根据本发明提供的详细公开内容。
例如,可以采用评估实体瘤和软瘤以及增殖性疾病或相关症状严重程度的任何体内或体外方法。
对于本发明方法中使用的任何制剂,治疗有效量或剂量可以最初从体外和细胞培养测定中评估。例如,可以在动物模型中配制剂量以达到所需的浓度或效价。此类信息可用于更准确地确定人体的有用剂量。
本发明所述的活性成分的毒性和治疗功效可以通过体外、在细胞培养基或实验动物中的标准药学程序确定。从这些体外和细胞培养测定以及动物研究中获得的数据可用于制定用于人类的一系列剂量。剂量可以根据使用的剂型和使用的给药途径而变化。个体医生可以根据患者的状况选择确切的配方、给药途径和剂量。(参见例如,Fingl,et al.,1975,in"The Pharmacological Basis ofTherapeutics",Ch.1p.1)。
可以单独调整剂量和间隔以提供足够量的活性成分以诱导或抑制生物效应(最小有效浓度,MEC)。每种制剂的MEC会有所不同,但可以根据体外数据进行估算。达到MEC所需的剂量将取决于个体特征和给药途径。检测分析可用于确定血浆浓度。
根据待治疗病症的严重性和反应性,给药可以是单次或多次给药,治疗过程持续数天至数周或直至达到治愈或实现疾病状态的减轻。
当然,要施用的组合物的量将取决于被治疗的对象、疾病的严重程度、施用的方式、处方医师的判断等。
如果需要,本发明的一些实施例的组合物可以存在于包装或分配装置中,例如FDA批准的试剂盒,其可以包含一种或多种含有活性成分的单位剂型。例如,该包装可以包括金属或塑料箔,例如泡罩包装。包装或分配装置可以附有给药说明。包装或分配器也可以通过与容器相关联的通知以由管理药品的制造、使用或销售的政府机构规定的形式来适用,该通知反映了该机构对组合物形式或人用给药或兽用给药的批准。例如,此类通知可能是美国食品和药物管理局批准的处方药标签或批准的产品内附文件。如上文进一步详述的,还可以制备包含配制在相容的药物载体中的本发明制剂的组合物,将其放置在合适的容器中,并标记用于治疗的指定病症。
在另一个实施例中,本发明提供食物或饮料形式的营养或膳食组合物,其包含本发明所述的组分。这些食物或饮料包含本发明组合物的各种示例性实施方式。这些食品或饮料可以制成或提供为谷物、婴儿食品、健康食品或特定健康用途的食品,例如巧克力或营养棒等固体食品、奶油或果酱等半固体食品或凝胶;也可为饮料。此类食品或饮料项目的具体和非限制性实施例包括清凉饮料、乳酸菌饮料、滴剂、糖果、口香糖、巧克力、软糖、酸奶、冰淇淋、布丁、软红豆果冻、果冻、饼干等。
在本发明的其他实施例中,所述组合物的组分是本发明提及的植物产物和提取物的合成类似物。
本教导进一步设想用其他抗病毒药物或抗炎药物或抗凝血剂作为单独的治疗或在联合制剂中进行治疗。
不限于实体瘤和软瘤以及增殖性疾病,但为了举例,根据一个具体实施例,抗病毒药物选自瑞德西韦、干扰素、利巴韦林、阿德福韦、替诺福韦、阿昔洛韦、布立夫定、西多福韦、福米韦森、膦甲酸、更昔洛韦、喷昔洛韦、金刚烷胺、金刚乙胺和扎那米韦。
如本发明所用,术语“约”是指±10%。
术语“包括”、“包含”、“具有”及其共轭词意为“包括但不限于”。
术语“由……组成”是指“包括并限于”。
术语“基本上由…组成”是指组合物、方法或结构可以包括附加的成分、步骤和/或部分,但前提是附加的成分、步骤和/或部分不会实质性地改变所要求保护的组合物、方法或结构的基本和新颖特征。
如本发明所用,单数形式“a”、“an”和“the”包括复数引用,除非上下文另有明确规定。例如,术语“一种化合物”或“至少一种化合物”可以包括多种化合物,包括其混合物。
在本申请全文中,本发明的各种实施方式可以以范围格式呈现。应当理解,范围格式的描述仅仅是为了方便和简洁,不应理解为对本发明范围的非灵活性限制。因此,范围的描述应该被认为已经具体公开了所有可能的子范围以及该范围内的各个数值。例如,对诸如1到6的范围的描述应该被认为具有具体公开的子范围,例如从1到3、从1到4、从1到5、从2到4、从2到6、从3到6等,以及该范围内的单个数字,例如1、2、3、4、5和6。无论范围的宽度如何,这都适用。
无论何时在本发明中指出数值范围,其旨在包括在所指出的范围内的任何引用的数字(分数或整数)。短语“范围/范围在”第一指示数字和第二指示数字“之间”和“范围/范围从”第一指示数字“到”第二指示数字在本发明中可互换使用并且旨在包括第一和第二指示数字以及它们之间的所有小数和整数。
如本发明所用,术语“方法”是指用于完成给定任务的方式、手段、技术和程序,包括但不限于化学、药理学、生物、生化和医学领域的从业者已知的或易于从已知方式、手段、技术和程序开发的。
如本发明所用,术语“治疗”包括消除、实质上抑制、减缓或逆转病状的进展,实质上改善病状的临床或美学症状或实质上防止病状的临床或美学症状的出现。
应当理解,为了清楚起见,在单独实施方式的上下文中描述的本发明的某些特征也可以在单个实施方式中组合提供。相反,为了简洁起见,在单个实施方式的上下文中描述的本发明的各种特征也可以在本发明的任何其他描述的实施方式中单独或以任何合适的子组合或适当的提供。在各种实施方式的上下文中描述的某些特征不应被认为是那些实施方式的基本特征,除非实施方式在没有这些元素的情况下是无效的。
如上文所描述和如在以下权利要求部分中所要求保护的本发明的各种实施例和方面在以下实施例中找到了实验支持。
实施例
现在参考以下实施例,其与以上描述一起以非限制性方式说明本发明的一些实施方式。
通常,本发明使用的命名法和本发明使用的实验室程序包括分子、生化、微生物和重组DNA技术。这些技术在文献中得到了详尽的解释。参见,例如,"Molecular Cloning:Alaboratory Manual"Sambrook et al.,(1989);"Current Protocols in MolecularBiology"Volumes I-III Ausubel,R.M.,ed.(1994);Ausubel et al.,"CurrentProtocols in Molecular Biology",John Wiley and Sons,Baltimore,Maryland(1989);Perbal,"A Practical Guide to Molecular Cloning",John Wiley&Sons,New York(1988);Watson et al.,"Recombinant DNA",Scientific American Books,New York;Birren et al.(eds)"Genome Analysis:A Laboratory Manual Series",Vols.1-4,ColdSpring Harbor Laboratory Press,New York(1998);methodologies as set forth inU.S.Pat.Nos.4,666,828;4,683,202;4,801,531;5,192,659and 5,272,057;"CellBiology:A Laboratory Handbook",Volumes I-III Cellis,J.E.,ed.(1994);"CultureofAnimal Cells-A Manual of Basic Technique"by Freshney,Wiley-Liss,N.Y.(1994),Third Edition;"Current Protocols in Immunology"Volumes I-III Coligan J.E.,ed.(1994);Stites et al.(eds),"Basic and Clinical Immunology"(8th Edition),Appleton&Lange,Norwalk,CT(1994);Mishell and Shiigi(eds),"Selected Methods inCellular Immunology",W.H.Freeman and Co.,New York(1980);可用的免疫测定在专利和科学文献中被广泛描述,参见,例如,美国专利号3,791,932;3,839,153;3,850,752;3,850,578;3,853,987;3,867,517;3,879,262;3,901,654;3,935,074;3,984,533;3,996,345;4,034,074;4,098,876;4,879,219;5,011,771和5,281,521;"OligonucleotideSynthesis"Gait,M.J.,ed.(1984);“Nucleic Acid Hybridization"Hames,B.D.,andHiggins S.J.,eds.(1985);"Transcription and Translation"Hames,B.D.,and HigginsS.J.,eds.(1984);"Animal Cell Culture"Freshney,R.I.,ed.(1986);"ImmobilizedCells and Enzymes"IRL Press,(1986);"A Practical Guide to Molecular Cloning"Perbal,B.,(1984)and"Methods in Enzymology"Vol.1-317,Academic Press;"PCRProtocols:A Guide To Methods And Applications",Academic Press,San Diego,CA(1990);Marshak et al.,"Strategies for Protein Purification andCharacterization-A Laboratory Course Manual"CSHL Press(1996);所有这些都通过引用并入本发明,如同在本发明中完全阐述一样。在本发明全文中提供了其他一般性引用。其中的程序被认为是本领域公知的并且是为了读者的方便而提供的。其中包含的所有信息通过引用并入本发明。
实施例1
用于治疗实体瘤和软瘤以及增殖性疾病的试验
除了传统的体内动物模型外,还有许多基于细胞的体外系统用于评估实体瘤和软瘤以及增殖性疾病的生长和治疗反应性。为了模拟实体瘤和软瘤以及增殖性疾病,培养来自各种组织的肿瘤的原代细胞系和细胞系,然后将其暴露于治疗的组合物和/或组分。在存在或不存在添加的本发明的组合物和/或组分的情况下测定细胞反应,特别是增殖、衰老和代谢活性,以评估本发明的组合物或组分减少或以其他方式改变肿瘤表型的能力。细胞可以在2-D或3-D培养基中繁殖。
用于几乎所有器官和组织的实体瘤和软瘤以及增殖性疾病的体外建模的示例性细胞类型在商业上广泛可获得,例如用于肾上腺肿瘤的NCI-H295R细胞,用于膀胱肿瘤的HT-1376、J82、T24P细胞,用于脑肿瘤的DBTRG、LN-18、SF-295、SF-767和SNB-19细胞,用于宫颈肿瘤的Ca Ski、He La和KB细胞,用于结肠癌的COLO 205、DLD-1、HCT、LoVo和NCI-H508细胞,用于上皮肿瘤的HEKn细胞,用于食管肿瘤的OE33细胞,用于尤文氏肉瘤(Ewingssarcoma)的A4573细胞,用于成纤维细胞衍生肿瘤的NHDF和Hs 895T细胞,用于胃肿瘤的GIST-T1和NCI-N87细胞,用于头部和颈部肿瘤的CAL 27细胞,用于肝脏肿瘤的Hep、Hepa和BLN细胞,用于肺部肿瘤的Calu-6、NCI-H596、NCI-H125-Luc、HCC827、LL和LL/2细胞,用于淋巴瘤的YAC-1、DB、GRANTA-519、EBC-1、Daudi、Raji和RL细胞,用于乳腺肿瘤的HCC70、MCF-7、MDA-MB、SK-BR3和MX-1细胞,用于黑色素瘤的SK-MEL和OCM细胞,用于间皮瘤的AB1细胞,用于骨髓瘤的RPMI 8226和OP M-2细胞,用于神经母细胞瘤的SK-N-F1,用于卵巢癌的OVCAR细胞,用于胰腺癌的PANC-1和Capan细胞,用于前列腺癌的PC-3和VCaP细胞,用于肾癌的ACHN和Renca细胞,用于肉瘤的MG-63、A-673和SW 872细胞,用于甲状腺肿瘤的TT和MB-1细胞,用于外阴癌的SK-LMS细胞。
肿瘤细胞可以在体外进行评估,有些可以通过引入动物用于异种移植生长试验。在一个示例性实施例中,来自原代培养物或细胞系的肿瘤细胞在体外培养,或者注射到循环中、皮下注射或者直接注射到小鼠或大鼠的靶器官中,并且肿瘤的建立和它们的生长可以是通过直接测量或检测标记细胞进行监测。动物宿主可以是具有免疫能力或免疫缺陷(SCID,裸)。本发明的组合物和/或组分的抗肿瘤功效可以在肿瘤细胞生长的多个阶段—通过在体外细胞生长阶段(移植前)给药、在引入宿主动物阶段、以及在异种移植肿瘤被允许在宿主动物中达到一定大小后对已建立的异种移植肿瘤的影响进行评估和评价。
实体瘤和软瘤以及增殖性疾病生长的动物模型
实体瘤和软瘤以及增殖性疾病的动物模型包括诱导动物模型、转基因模型和自然发生的过度增殖性疾病和病症的动物模型。
除了上文提到的动物模型外,基因工程小鼠和大鼠癌症提供了强大的肿瘤体内模型,允许有机会评估肿瘤在其自然环境中的药物递送、治疗反应和生物标志物表达。适于评估本发明的组合物和组分的功效的基因工程动物模型包括但不限于:MMTV-PyMT小鼠乳腺肿瘤基因工程小鼠、用于鳞状皮肤肿瘤的K14-HPV16小鼠和KB1P乳腺癌小鼠模型。
尽管已经结合其特定实施方式描述本发明,但显然许多替代、修改和变化对于本领域技术人员来说将是显而易见的。相应地,旨在囊括落入所附权利要求的精神和广泛范围内的所有这些替代、修改和变化。
申请人的目的是,本说明书中提及的所有出版物、专利和专利申请都将通过引用整体并入本说明书中,就好像每个单独的出版物、专利或专利申请在引用时是具体和单独注明将通过引用整体并入本发明。此外,本申请中任何参考文献的引用或标识不应被解释为承认此类参考文献可用作本发明的现有技术。就使用章节标题而言,它们不应被解释为必然限制。此外,本申请的任何优先权文件在此通过引用整体并入本发明。
实施例2
鳞状细胞癌(SCC)和基底细胞肉瘤(BCC)的治疗
皮肤鳞状细胞癌(SCC)是第二种最常见的皮肤癌形式,其特征是鳞状细胞异常、加速生长。早期发现时,大多数SCCs是可以治愈的。鳞状细胞是位于皮肤表面附近的扁平细胞,随着新细胞的形成会不断脱落。SCCs可表现为鳞片状红色斑块、开放性溃疡、粗糙、增厚或疣状皮肤,或具有中央凹陷的凸起生长。有时,SCCs可能会结痂、发痒或流血。病变最常见于身体暴露在阳光下的区域。基底细胞癌(BCC)是最常见的皮肤癌形式,也是所有癌症中最常见的形式。BCCs源于基底细胞的异常、不受控制的生长,基底细胞是皮肤顶层的三种主要细胞类型之一。BCCs可能看起来像开放性溃疡、红色斑块、粉红色生长物、闪亮的肿块、疤痕或具有略微隆起、卷边和/或中央凹痕的生长物。有时,BCCs可能会渗出、结痂、发痒或流血。病变通常出现在身体暴露在阳光下的区域。由于生长缓慢,大多数BCCs是可治愈的,并且在早期发现和治疗时造成的损害最小。SCC和BCC最常发生在因暴露于太阳紫外线(UV)辐射造成DNA损伤时(参见skincancer.org)。
一名有BCC病史的患者在3次活检阳性后被诊断为位于左耳的SCC。还注意到疑似BCC受累,但没有淋巴结受累。在左耳的肿瘤旁边,面部的变色和色素沉着很明显。皮肤癌患者在当前治疗前被诊断为BCC和侵入性SSC。
在本发明的抗癌治疗后,在超声和CT检查中均发现所有组织均清晰且未涉及淋巴结。
用本发明组合物处理三个月后。描述面部的变色和色素沉着被根除。如图2和图3所示。
本发明所述的方法、疫苗、药物组合物、组合物或食品补充剂,其中,所述实体瘤和软瘤以及增殖性疾病选自包含肉瘤和癌的组,例如纤维肉瘤、粘液肉瘤、脂肪肉瘤、软骨肉瘤、成骨肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤、尤文氏瘤、平滑肌肉瘤、横纹肌肉瘤、结肠癌、胰腺癌、乳腺癌、卵巢癌、前列腺癌、鳞状细胞癌、基底细胞癌、腺癌、汗腺癌、皮脂腺癌、乳头状癌、乳头状腺癌、囊腺癌、髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎癌、威尔姆氏瘤、宫颈癌、睾丸癌、肺癌、小细胞肺癌、膀胱癌、上皮癌、多形性胶质母细胞瘤、星形细胞瘤、髓母细胞瘤、颅咽管瘤、室管膜瘤、松果体瘤、血管母细胞瘤、听神经瘤、少突胶质细胞瘤、皮肤T细胞淋巴瘤(CTCL)、皮肤B细胞淋巴瘤(CBCL)、黑色素瘤、神经母细胞瘤、视网膜母细胞瘤、霍奇金淋巴瘤、非霍奇金淋巴瘤、弥漫性大B细胞淋巴瘤、慢性淋巴性白血病、套细胞淋巴瘤、滤泡性淋巴瘤、脾边缘区淋巴瘤、淋巴结边缘区淋巴瘤、结外边缘区淋巴瘤、伯基特淋巴瘤(Burkitt's Lymphoma)、浆母细胞淋巴瘤、外周T细胞淋巴瘤非特指型(Peripheral TcellLymphomaNOS)、毛细胞白血病(HCL)、急性淋巴细胞白血病(ALL)、急性髓细胞白血病(AML)、急性早幼粒细胞白血病(APL)、慢性淋巴细胞白血病(CLL)、慢性髓细胞白血病(CML)、骨髓增生性肿瘤(MPN)和系统性肥大细胞增多症、乳头状甲状腺癌、非侵袭性滤泡性甲状腺肿瘤、滤泡性甲状腺癌、癌、甲状腺髓样癌、甲状腺未分化癌、甲状腺淋巴瘤、鳞状细胞甲状腺癌、甲状腺肉瘤、何氏细胞癌(Hürthle Cell Carcinoma)。
实施例3
用本发明组合物治疗患有结肠癌的妇女。CT和超声波检查显示在用本发明的组合物治疗13天后肿瘤减少并且在一些地方肿瘤消失。
本发明并不旨在限制于上述说明和描述的实施例,而是可以在由以下权利要求限定的本发明的范围和精神内进行修改和变化。
附录
不同地理来源的百里香精油的化学组分比较
A.拉尔1,E.阿拉克1,A.奥拉夫2
1药学研究所,塔尔图大学
努鲁斯街.1,50411塔尔图,爱沙尼亚
2化学研究所,塔林理工大学
埃希塔加特座5,19086塔林(Tallinn),爱沙尼亚
摘要
使用毛细管气相色谱分析方法确定了在爱沙尼亚和其他欧洲国家种植的百里香精油组分的变化。鉴定出59种组分,占总油产量的95%以上。普通百里香油中的主要组分是百里酚(0.9%-75.7%)、香芹酚(1.5%-83.5%)、对伞花烃(4.3%-34.4%)、γ-松油烯(0.9%-19.7%)、芳樟醇(0.4%-4.8%)、(E)-β-石竹烯(0.5%-9.3%)和松油烯-4-醇(tr.-3.8%)。所研究的油中酚类化合物(百里酚和香芹酚)的总和从19.4%到84.4%不等,其前体(对伞花烃和γ-松油烯)的总和从5.7%到38.5%不等。荷兰(65.5%)和爱沙尼亚(75.7%)的油中百里酚含量占主导地位,而希腊百里香油(83.5%)中香芹酚含量占主导地位。亚美尼亚百里香油中百里酚的含量仅为17.0%,但富含柠檬醛和香茅醇(32.5%)、冰片(4.3%)、香茅醛(4.0%)、1,8-桉树脑(4.0%)和甲基丁香酚和醋酸百里酚(7.5%)。在爱沙尼亚,普通百里香的百里酚、百里酚-香芹酚和百里酚-对伞花烃-γ-松油烯的化学型是可区分的。
关键词:银斑百里香L.、唇形科、普通百里香、精油、不同地理来源、百里酚、香芹酚、对伞花烃、γ-松油烯
百里香属内有许多种和亚种,其中大部分包括银斑百里香L.在内以百里酚和香芹酚为主要组分,而1,8-桉树脑、樟脑、柠檬醛、香芹酮、单萜醇以及乙酸酯和倍半萜醇的浓度则存在差异[1-14]。这些化学型,尤其是富含酚萜类(phenolic terpenoids)化合物,表现出很强的抗氧化活性[15,16]。在爱沙尼亚,已知的百里香属物种只有两种。普通百里香(Thymus vulgaris L.)种植以及野生百里香(Thymus serpyllum L.)生长在野外。对源自爱沙尼亚不同自然生长地的野生百里香精油组分的研究表明,至少存在三种化学型[17]。与其他国家的文献数据相反,百里酚和香芹酚不是爱沙尼亚野生百里香油的主要组分。
在目前的工作中,我们使用来自不同欧洲国家的商业普通百里香样品和在爱沙尼亚种植的样品确定了精油的成分。研究了生物活性成分含量的差异。来自爱沙尼亚的主要百里香油组分的浓度与其他欧洲国家的样品进行了比较。
材料和方法
植物材料(商业百里香)于2000年(法国)、2001年(匈牙利、荷兰)、2002年(俄罗斯、希腊、爱沙尼亚)和2003年(苏格兰、摩尔达维亚、亚美尼亚)从欧洲各国的零售药店获得。爱沙尼亚样品于2001、2002和2003年夏天从爱沙尼亚的不同生长地采集。凭证标本已存放在药学研究所(the Institute of Pharmacy),塔尔图大学(University ofTartu),爱沙尼亚。
毛细管气相色谱法
通过欧洲药典[18]中描述的蒸馏方法从普通百里香干草本植物中分离得到精油。使用带有FID的Chrom-5色谱仪在两根熔融石英毛细管柱(50m×0.20mm i.d.)上使用非极性聚二甲基硅氧烷(NB-30)和极性聚乙二醇20M(NB-20M)的固定相(Nordion,芬兰)分析精油。两种固定相的膜厚均为0.25μm。以氦气为载气,分流率为1:150,流速为20-25cm/秒。温度程序以2℃/分钟速度从50-250℃,进样器温度为250℃。3390A Hewlett-Packard积分器用于数据处理。
气相色谱-质谱联用仪
气相色谱-质谱联用仪(GC-MS)数据在Hewlett-Packard 5988A仪器上获得。MS条件如下:El模式70ev,离子源温度200℃。GC条件为以5℃/分钟速度从60-280℃,内部保持时间为2分钟。使用氦气作为载气,流速为20cm/秒。熔融石英毛细管柱AT-5,聚(5%-苯基-95%-二甲基硅氧烷),使用(25m×0.25mm i.d.,膜厚0.25μm)。进样器温度为280℃。
鉴定和定量评价
通过比较NB-30和NB-20M色谱柱上GC峰的保留指数(RI)与标准化合物的RI值、我们的RI数据库和文献[19-21]来鉴定化合物。所得结果通过GC-MS得到证实。油的定量组成根据NB-30色谱柱上的GC峰面积计算,没有FID响应因子校正,使用归一化方法。
结果与讨论
银斑百里香L.的精油组分在两根不同极性色谱柱的RI值、爱沙尼亚和其他欧洲国家的百里香油组分百分比见表1。
表1.
不同来源的银斑百里香L.精油的组成(composition),%。
产量最高的组分以粗体显示;tr-痕量(<0.05%),
*-暂时鉴定。
在所研究的样品中鉴定出59种组分,占总油的95%以上。油中的主要化合物组是含氧单萜类化合物(oxygenated monoterpenoids)(40.4%-86.8%),包含酚类(百里酚和香芹酚):19.4%-84.4%。单萜构成油的8.3%-42.1%,包含酚类前体(对伞花烃和γ-松油烯):5.7%-38.5%。倍半萜占百里香油的0.3%-17.6%。油中的主要倍半萜是(E)-β-石竹烯(0.5%-9.3%)、大根香叶烯D(0%-4.3%)、β-红没药烯(0%-2.6%)和蛇床-3,7(11)-二烯(0%-2.4%)。其他倍半萜在所有样品中占1%不到。从百里香油中鉴定的含氧倍半萜中,仅发现氧化石竹烯(0.1%-2.5%)含量超过1%。
对来自不同地理来源样品的百里香油组成的比较表明,大多数生物活性成分存在一定的差异。在希腊产油中,香芹酚含量为83.5%。在其他所研究的样品中,该值从2.2%到4.1%不等。在爱沙尼亚和荷兰的两个百里香样品中,油中的百里酚含量(分别为75.7%、67.5%和65.5%)高于其他样品(0.9-49.0%)。酚类前体对伞花烃和γ-松油烯的浓度总和,从5.7%到38.5%不等,这些值在亚美尼亚(5.7%)和希腊(7.8%)的油中最低。所研究的百里香油中的四种主要成分(百里酚、香芹酚、对伞花烃和γ-松油烯)的含量范围为67.7%至92.2%。唯一的例外是来自亚美尼亚的油,该值仅占25.1%。亚美尼亚百里香油中富含柠檬醛和香茅醇(32.5%)、甲基丁香酚和乙酸百里酚(7.5%)、冰片(4.3%)、香茅醛(4.0%)和1,8-桉树脑(4.0%)。
如表2所示,爱沙尼亚样品6和7中的百里酚化学型清晰可辨(百里酚含量分别为75.7%和67.5%)。样品4、8和10富含百里酚(22.5%-45.1%)和香芹酚(29.9%-34.6%),而样品1、2、3和5富含百里酚(41.7%-49.0%)和对伞花烃(14.6%-22.2%)。与所研究的其他油不同,样品9含有的百里酚、香芹酚和对伞花烃相对较少(共45.6%),但富含单萜(月桂烯-5.1%)和倍半萜(β-石竹烯-9.3%,大根香叶烯D-4.3%)。
就来自不同地理来源的普通百里香油中的生物活性化合物而言,这项工作的结果已经建立了明显的数量差异。因此,这些药用植物具有基本的抗菌和抗菌性质,它们的药理作用也可能不同。
荷兰的油和爱沙尼亚的两种油属于百里酚化学型,而法国、匈牙利、俄罗斯和苏格兰的油属于富含百里酚-对伞花烃的化学型。仅在爱沙尼亚,百里酚-香芹酚和百里酚-对伞花烃-γ-松油烯化学型是可区分的。来自希腊的油被发现是一种富含香芹酚的化学型。与其他油不同,来自亚美尼亚的油含有大量的柠檬醛和香茅醇。
表2.来自不同地理来源的百里香主要精油组分的浓度。
结论
来自不同地理来源的普通百里香精油的主要组分是百里酚、香芹酚、对伞花烃、γ-松油烯、芳樟醇、(E)-β-石竹烯和松油烯-4-醇。
在爱沙尼亚,普通百里香的百里酚、百里酚-香芹酚和百里酚-对伞花烃-γ-松油烯化学型是可区分的。
致谢
爱沙尼亚科学基金会(第4332号赠款)为此处报告的工作提供了财政支持。
kerla xolouea
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不同地理来源的百里香精油的化学组分比较
A.拉尔1,E.阿拉克1,A.奥拉夫2
1药学研究所,塔尔图大学
努鲁斯街.1,50411塔尔图,爱沙尼亚
2化学研究所,塔林理工大学
埃希塔加特座5,19086塔林(Tallinn),爱沙尼亚
摘要
使用毛细管气相色谱分析方法确定了在爱沙尼亚和其他欧洲国家种植的百里香精油组分的变化。鉴定出59种组分,占总油产量的95%以上。普通百里香油中的主要组分是百里酚(0.9%-75.7%)、香芹酚(1.5%-83.5%)、对伞花烃(4.3%-34.4%)、γ-松油烯(0.9%-19.7%)、芳樟醇(0.4%-4.8%)、(E)-β-石竹烯(0.5%-9.3%)和松油烯-4-醇(tr.-3.8%)。所研究的油中酚类化合物(百里酚和香芹酚)的总和从19.4%到84.4%不等,其前体(对伞花烃和γ-松油烯)的总和从5.7%到38.5%不等。荷兰(65.5%)和爱沙尼亚(75.7%)的油中百里酚含量占主导地位,而希腊百里香油(83.5%)中香芹酚含量占主导地位。亚美尼亚百里香油中百里酚的含量仅为17.0%,但富含柠檬醛和香茅醇(32.5%)、冰片(4.3%)、香茅醛(4.0%)、1,8-桉树脑(4.0%)和甲基丁香酚和醋酸百里酚(7.5%)。在爱沙尼亚,普通百里香的百里酚、百里酚-香芹酚和百里酚-对伞花烃-γ-松油烯的化学型是可区分的。
关键词:银斑百里香L.、唇形科、普通百里香、精油、不同地理来源、百里酚、香芹酚、对伞花烃、γ-松油烯
Claims (22)
1.一种在有需要的受试者中预防或治疗实体瘤和软瘤以及增殖性疾病的方法,其特征在于,所述方法包含向受试者施用有效量的植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物种选自一类包括黑种草Nigella sativa、头状百里香Thymus capitatus、银斑百里香Thymus vulgaris、牛至Origanum syriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamumindicum和漆树Rhus coriaria,绞股蓝Gynostemma petaphyllum、乳香和人参的组;所述方法用于预防或治疗受试者的实体瘤和软瘤和增殖性疾病。
2.一种针对实体瘤和软瘤以及增殖性疾病的疫苗,其特征在于,其包含有效量的的植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗或预防实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草Nigella sativa、头状百里香Thymuscapitatus、银斑百里香Thymus vulgaris、牛至Origanum syriacum、百里香Thymbraspicata、百里香Satujera thymbra、芝麻Sesamum indicum和漆树Rhus coriaria,绞股蓝Gynostemma petaphyllum、乳香和人参的组。
3.一种药物组合物,其特征在于,其包含有效量的植物种或其属衍生的组分,选择含有植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗或预防实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草Nigella sativa、头状百里香Thymus capitatus、银斑百里香Thymusvulgaris、牛至Origanum syriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamum indicum和漆树Rhus coriaria,绞股蓝Gynostemma petaphyllum、乳香和人参的组;所述药物组合物用于预防或治疗实体瘤和软瘤以及增殖性疾病。
4.一种物质组合物,其特征在于,其包含至少2种植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、其模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草Nigella sativa、头状百里香Thymus capitatus、银斑百里香Thymusvulgaris、牛至Origanum syriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamum indicum和漆树Rhus coriaria,绞股蓝Gynostemma petaphyllum、乳香和人参的组。
5.一种食品补充剂,其特征在于,其包含至少2种植物物种或其属衍生的组分,选自一类含有植物部分、其提取物、其级分、其活性成分、其合成类似物、模拟物或其组合物的组,其中,所述组分能够治疗实体瘤和软瘤以及增殖性疾病,并且其中所述植物物种选自一类包括黑种草Nigella sativa、头状百里香Thymus capitatus、银斑百里香Thymusvulgaris、牛至Origanum syriacum、百里香Thymbra spicata、百里香Satujera thymbra、芝麻Sesamum indicum和漆树Rhus coriaria,绞股蓝Gynostemma petaphyllum、乳香和人参的组。
6.如权利要求1-5之任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述实体瘤和软瘤以及增殖性疾病,选自一类含有肉瘤和癌的组,包括纤维肉瘤、粘液肉瘤、脂肪肉瘤、软骨肉瘤、成骨肉瘤、脊索瘤、血管肉瘤、内皮肉瘤、淋巴管肉瘤、淋巴管内皮肉瘤、滑膜瘤、间皮瘤、尤文氏瘤、平滑肌肉瘤、横纹肌肉瘤、结肠癌、胰腺癌、乳腺癌、卵巢癌、前列腺癌、鳞状细胞癌、基底细胞癌、腺癌、汗液腺癌、皮脂腺癌、乳头状癌、乳头状腺癌、囊腺癌、髓样癌、支气管癌、肾细胞癌、肝细胞癌、胆管癌、绒毛膜癌、精原细胞瘤、胚胎癌、威尔姆氏瘤、宫颈癌、睾丸癌、肺癌、小细胞肺癌、膀胱癌、上皮癌、多形性胶质母细胞瘤、星形细胞瘤、髓母细胞瘤、颅咽管瘤、室管膜瘤、松果体瘤、血管母细胞瘤、听神经瘤、少突胶质细胞瘤、皮肤T细胞淋巴瘤(CTCL)、皮肤B细胞淋巴瘤(CBCL)、黑色素瘤、神经母细胞瘤、视网膜母细胞瘤、霍奇金淋巴瘤、非霍奇金淋巴瘤、弥漫性大B细胞淋巴瘤、慢性淋巴性白血病、套细胞淋巴瘤、滤泡性淋巴瘤、脾边缘区淋巴瘤、淋巴结边缘区淋巴瘤、结外边缘区淋巴瘤、伯基特淋巴瘤、浆母细胞淋巴瘤、外周T细胞淋巴瘤NOS、毛细胞白血病(HCL)、急性淋巴细胞白血病(ALL)、急性髓细胞白血病(AML)、急性早幼粒细胞白血病(APL)、慢性淋巴细胞白血病(CLL)、慢性髓细胞白血病(CML)、骨髓增生性肿瘤(MPN)和系统性肥大细胞增多症、甲状腺乳头状癌、非侵袭性甲状腺滤泡性肿瘤、滤泡性甲状腺、癌症、甲状腺髓样癌、甲状腺未分化癌、甲状腺淋巴瘤、鳞状细胞甲状腺癌、甲状腺肉瘤、何氏细胞癌。
7.如权利要求1-5之任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述实体瘤、软瘤和增殖性疾病是脑癌、乳腺癌、三阴性乳腺癌、膀胱癌、骨癌、结直肠癌、肺癌、肾癌、肝癌、胃癌、前列腺癌、肉瘤、黑色素瘤、癌或淋巴瘤、纤维瘤fibroids、子宫内膜异位症。
8.如权利要求1-5之任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述实体瘤和软瘤以及增殖性疾病是淋巴瘤。
9.如权利要求1-3之任一项所述的方法、疫苗或药物组合物,其特征在于,所述组分包含至少2种组分。
10.如权利要求4-5、9中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含至少3种组分。
11.如权利要求4-5、9中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含至少4种组分。
12.如权利要求4-5、9中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含至少5种组分。
13.如权利要求4-5、9中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含5-10种组分。
14.如权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含百里醌或其类似物。
15.根据权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含百里酚或其类似物。
16.根据权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,其中,所述组分包含香芹酚或其类似物。
17.根据权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,所述组分包含菠萝蛋白酶或其类似物。
18.方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,其包含菠萝提取物,所述菠萝提取物包含菠萝蛋白酶或其类似物。
19.根据权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,进一步包含大麻或大麻素。
20.根据权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,进一步包含色氨酸。
21.如权利要求1-13中任一项所述的方法、疫苗、药物组合物、物质组合物或食品补充剂,其特征在于,进一步包括食品补充剂、物质组合物或提取物;
进一步包括“贝都因茶”,其中,所述“贝都因茶”包括玫瑰叶姜味草灌木Rose LeavesMicromeria fruticose、鼠尾草Salvia、柠檬草(cymbopgon)(柠檬醛)、阿莱藤属Aloysia、马鞭草verbena officinalis、马郁兰origanum majorana、薄荷。
22.如权利要求1-中任一项所述的方法、疫苗、药物组合物、物质组合物、提取物或食品补充剂,其特征在于,其进一步包括“贝都因茶”,其中,所述“贝都因茶”包括百里香、鼠尾草、小豆蔻、肉桂、红茶、哈布克Habuk、马尔马亚Marmaya。
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