CN101932240A - 抗糖化方法及组合物 - Google Patents
抗糖化方法及组合物 Download PDFInfo
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- CN101932240A CN101932240A CN2008801179172A CN200880117917A CN101932240A CN 101932240 A CN101932240 A CN 101932240A CN 2008801179172 A CN2008801179172 A CN 2008801179172A CN 200880117917 A CN200880117917 A CN 200880117917A CN 101932240 A CN101932240 A CN 101932240A
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Abstract
本发明包括提供抗糖化活性的组合物,该组合物包括矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物。所述组合物用于预防、治疗和抑制体内的糖化效应的方法中。本发明的方法包括抗糖化组合物在对包括糖尿病、动脉粥样硬化、关节炎、精神症状和视力障碍的与糖化相关的症状的治疗和预防中的用途。
Description
相关申请
本发明要求2007年10月10日提交的美国临时专利申请第60/998,316号和2008年7月11日提交的美国临时专利申请第61/079,826号的优先权,每个申请的全部内容在此并入。
技术领域
本发明涉及用于治疗或预防人类和动物的糖化事件及相关病变的组合物。更特别的是,本发明涉及具有抗糖化活性的组合物。
发明背景
糖化或非酶糖化包括蛋白质、脂质或核酸的氨基与糖的醛基或酮基反应以产生修饰的氨基并且最终生成高级糖化终产物(AGE)。糖化可为已知为Amadori反应、Schiff碱反应和Maillard反应的一系列体内慢反应的第一步,并且产生高级糖化终产物(AGE)。尽管糖化在体内进行缓慢,但是糖化产物可具有反应性或可具有长期持续的效应,并且AGE的出现与衰老和病理状态相关。在其中存在大量可用于反应的糖分子的生理状态下,例如那些诸如糖尿病的糖水平增高的症状,糖化效应可更为显著。一些AGE为良性的,但是其它的更具反应性,并且牵涉许多衰老相关的慢性疾病,所述疾病例如II型糖尿病、心血管疾病、阿耳茨海默病、癌症、外周神经病和诸如耳聋和失明的其它感觉缺失。糖化也在脂质和RNA/DNA修饰中起作用。
糖化的底物从体内缓慢清除,因为肾清除率仅为约30%。这一事实用于提供测定糖尿病中糖水平的方法。红细胞具有120天的寿命并且简单地可用于测量近期存在的糖化产物增加。糖化的血红蛋白水平(也称作HbAlc)被确定,并指示发生于人体的糖化水平。
目前,氨基胍通过与糖反应并阻断Amadori反应而用于减缓糖化。氨基胍可体外和体内还原葡萄糖衍生的AGE。亲核肼化合物氨基胍,目前被研究用作糖尿病中AGE并发症的治疗。抑制AGE形成或破坏已经形成的AGE的其他药物(例如,AGE阻断剂)也在积极研究中。例如,据报道AGE阻断剂用于导致收缩期高血压和严重心力衰竭改善的血管无弹性的逆转。
糖尿病是由缺乏胰岛素导致的代谢病症,并且通常通过血糖水平的增加而诊断。所述病症以胰岛素依赖性组织对葡萄糖摄入降低为特征。所述病症可通过胰岛素注射控制,但是长期的糖尿病并发症包括眼部病变(白内障形成和视网膜病)、肾病变(肾病)、神经元病变(神经病)和血管病变(血管病和动脉粥样硬化)。糖化在糖尿病相关的病变中起作用。
研究表明AGE可在动脉粥样硬化的发展中起作用。单核细胞具有AGE特异性受体(RAGE)并且被释放的细胞因子刺激而响应。血管壁的轻度损伤可暴露内皮下AGE,提高单核细胞的渗透并且引起动脉粥样硬化损伤的发展。脂蛋白循环也可以经受糖化,然后可以比非糖化脂蛋白更快的速率被上皮细胞吸收。
可抵抗AGE形成的长期效应以及预防或治疗AGE相关病变的组合物的摄入是有益的。在诸如饮料或增甜剂组合物的易于消耗的产品中提供所述化合物对于预防或治疗糖化相关症状是有利的。
概述
本发明包括用于糖化治疗和预防的方法和组合物。本发明的组合物包括用于抑制人类或动物的糖化反应,并预防和治疗糖化相关症状的矿物质提取物组合物。本发明的组合物包括在此描述的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物和罗汉果苷组合物。本发明的方法包括治疗或预防糖化相关症状的方法,包括提供有效量的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物以预防或减少发生于人类或动物的糖化事件。
矿物质提取物组合物、罗汉果苷/矿物质提取物组合物和/或罗汉果苷组合物,单独地或与其它组分组合地提供或施用于受治疗者(人类或动物)以形成抗糖化组合物。例如,本发明的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物可与食品或饮料结合以提供可被受治疗者消耗的抗糖化组合物。有效量的抗糖化组合物,例如,抑制诸如AGE的糖化产物的形成,并且预防或治疗体内与糖化事件相关的症状。所述糖化相关症状包括但不限于糖尿病、心血管疾病、阿耳茨海默病、癌症、外周神经病和诸如耳聋、肾功能不全和失明的其它感觉缺失。本发明的组合物包括与具有甜味或感觉具有甜味的化合物或组合物结合的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物以形成抗糖化增甜剂组合物。
附图说明
图1为显示三种矿物质提取物组合物在抗糖化测定中的结果的图。
图2A和2B为显示矿物质提取物组合物对线粒体代谢的效应的图。A:所有细胞群体中线粒体活性增加,B:基于每个细胞的线粒体活性增加。
图3为显示矿物质提取物组合物对人类细胞产生I型胶原的效应的图。
图4为显示体外矿物质提取物组合物对金属蛋白酶活性的图。
图5为显示本发明的组合物的胶原刺激活性的图。
图6A和6B为显示本发明的组合物的代谢刺激活性的图。
图7为显示本发明的组合物的糖化抑制活性的图。
详述
本发明包括治疗或预防与诸如糖化的蛋白质、脂质或核酸的糖化分子的存在相关的症状的并发症和病变的组合物和方法。糖化相关的症状包括在人体或动物体内由于糖化的蛋白质、脂质和核酸的存在引起的或与之相关的症状,并且包括但不限于炎性响应、动脉粥样硬化、阿耳茨海默病、糖尿病、癌症、心血管疾病和其它AGE-以及RAGE-相关的症状。AGE(高级糖化终产物)及其受体RAGE(高级糖化终产物的受体)牵涉如心血管疾病和糖尿病的多种症状。AGE引起多种长期细胞功能缺失和最终的疾病状态。实质上,糖-蛋白质复合物成为化学交联的,并且降低正常细胞的功能,同时促进自由基损伤。
本发明包括治疗或预防糖化相关症状的组合物和方法。本发明的组合物包括在此所述的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物和罗汉果苷组合物。本发明方法包括提供有效量的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物以预防或减少在受治疗者中发生的糖化事件,或者以治疗或预防人类或动物的糖化相关症状。包括矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的组合物在此被称为抗糖化组合物,但是该引用并非意在限制所述组合物的活性。
高级糖化终产物(AGE)已经涉及为影响健康、健康状态和寿命的关键因素,尤其是与炎症和其它疾病状态相关的关键因素。糖化为体内导致AGE形成的一系列复杂非酶反应的第一步,所述非酶反应已知为Amadori反应、Schiff碱反应和Maillard反应。某些反应性AGE或前体AGE的特征为蛋白质的共价交联的形成,这改变它们的结构和功能,并且被发现于细胞基质、基底膜、红细胞和血管壁组分中。AGE其它主要的特性涉及它们与多种细胞表面AGE-结合受体(RAGE)的相互作用,其导致活化和助氧化事件、促炎性事件。
大量证据表明AGE是几乎所有糖尿病并发症的重要的致病介质,所述糖尿病并发症通常被分为微血管病和大血管病。例如,在糖尿病患者的视网膜血管中发现AGE,并且AGE的水平与血清中AGE水平以及视网膜病的严重性相关。现在AGE涉及众多与年龄相关的退化,从冠状动脉疾病至皮肤老化。一些AGE为良性的,但是其它AGE比衍生它们的糖更具反应性,并且涉及许多与年龄相关的慢性疾病,诸如:糖尿病,其中β细胞被损伤;心血管疾病,其中内皮、纤维蛋白原和胶原被损伤;阿耳茨海默病,其中淀粉样蛋白是发展成AGE的反应的副产物;释放丙烯酰胺和其它副产物的癌症;外周神经病和其它感觉缺失,诸如由脱髓鞘作用产生的耳聋、肾功能不全和由于微血管损伤发生的失明。这一疾病的范围表明高级糖化终产物干扰整个身体的分子和细胞功能的作用的程度,以及与AGE相关的其它副作用的程度,所述副作用例如高度氧化的副产物,诸如过氧化氢的释放。
糖化底物从体内排出速度慢,并且肾清除率因子仅为约30%。结果,长寿命的细胞(如神经元)、长期作用的蛋白质(如眼部晶状体和胶原)和DNA可随着时间而累积严重的损伤。如肾小球中的代谢活性细胞、眼部的视网膜细胞和胰腺中的β细胞(产生胰岛素的)也具有受损的高度危险。血管上皮细胞被糖化直接损伤,这与动脉粥样硬化有关。动脉粥样硬化斑块易于在高血流区域累积,因为在这些位点,存在的糖分子和糖化终产物增加。糖化损伤产生血管壁中胶原的硬化,并且产生高血压。糖化也引起血管壁的削弱,这可引起微动脉瘤或大动脉瘤,其如果在脑部形成,可转而引起中风。
除了内源形成的AGE之外,AGE也可从外源来源导入体内。外源AGE可作为“饮食的”或“预形成的”AGE而被消耗。这些在当糖与蛋白质及脂肪一起烹调时形成。例如,烟草烟雾为公知的AGE的外源来源。吸烟时,烟草中的各种前-AGE的燃烧产生反应性的和有毒的AGE。血清AGE或LDL-联AGE在吸烟者体内显著升高。结果,据报道糖尿病吸烟者的动脉和眼部晶状体中显示较高的AGE沉积。
氨基胍,AGE形成的抑制剂,已经显示可在糖尿病动物中预防视网膜病。另外,AGE在糖尿病患者的外周神经中累积,并且抗-AGE剂的使用提高神经传导速率并且改善神经元血流异常。
动脉粥样硬化在糖尿病患者中显著被加速,并且伴随更高风险的心血管性死亡和脑血管性死亡。临床前研究和临床研究已经表明AGE在粥样硬化病变的形成和发展中起到重要作用。增加的AGE在糖尿病血管组织上的累积与上皮细胞、巨噬细胞和平滑肌细胞功能的变化相关。另外,AGE可以一种意在使LDL胆固醇易于被氧化并在管壁内沉积的方式修饰LDL胆固醇,导致划痕的形成,并且最后导致动脉粥样化。AGE-交联形成产生动脉硬化和大血管弹性的丧失。
AGE在正常体内以恒定但是缓慢的速率形成,起始于早期胚胎发育,并且随着时间累积。然而,他们的形成随着衰老或暴露于诸如葡萄糖、果糖和半乳糖的糖源的增加而显著加速。
令人惊奇的发现,在此所述的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物显示可与氨基胍相当或比其更好的抗糖化性质。矿物质涉及体内大量的复杂反应,大多数反应依赖于矿物质的化学特性。矿物质和电解质保持体液平衡和酸碱平衡,而且它们在许多酶催化反应中具有作为辅因子的化学活性。本发明的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物可包括干粉、液体制剂、食品组合物、化妆品组合物和通过注射施用的组合物。组合物可被添加于第二组合物中以形成结合的组合物,以将发现于矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物中的抗糖化活性提供给第二组合物。
本发明的组合物包括矿物质提取物组合物。生产所述组合物的方法教导于美国专利公布第2005/0118279号中,其全部内容在此并入。简要地,为了生产本发明的矿物质提取物组合物,收集来自合适的地点的包含描述于美国专利公开第2005/0118279号中的元素的土壤,并且使其进入其中描述的水溶性提取过程中以产生包含矿物质元素的液体矿物质元素组合物,该组合物可被干燥以产生干粉矿物质元素组合物。
液体矿物质提取物组合物和干粉矿物质提取物组合物包括描述于美国专利公开第2005/0118279号中的综合矿物质组合物中的矿物质元素。按在此描述的步骤生产的液体矿物质提取物组合物和干粉矿物质提取物组合物通常包含最少8种常量矿物质元素和最少60种微量矿物质元素。
也对液体矿物质提取物组合物和干燥矿物质提取物组合物进行物理学测试和分析。典型的液体矿物质提取物组合物的分类可以颜色分类并可为从黄色至琥珀褐色,并含有按重量计1%至10%之间的矿物质元素,或按重量计3-5%的矿物质元素。液体矿物质提取物组合物为酸性,具有2.5-4.5或2.5-3.5的pH范围。液体矿物质提取物组合物可被干燥以产生无水粉末。所述无水粉末的颜色范围可从淡米白色至褐色,或从黄色至金琥珀色,在诸如亲脂性液体(油和脂肪)的非极性溶剂中不溶,在醇中不溶,并且以按重量计1%至5%的浓度或按重量计3-5%的浓度在水和水醇(hydro-alcoholic)溶液中易溶,但不膨胀。干粉以过饱和溶液的形式部分溶解于极性溶剂中或能够部分悬浮于极性溶剂中。
以在此描述的步骤生产的液体矿物质提取物组合物和干燥矿物质提取物组合物可含有最少8种常量矿物质元素和最少60种微量矿物质元素。微量矿物质元素包括痕量和稀土族矿物质元素。
例如,干燥矿物质提取物组合物可含有常量矿物质元素钙、氯、镁、锰、磷、钾、硅和钠,浓度范围按重量百分比计从0.0001-20.00%,按重量百分比计从0.001%-10%、从0.1%至20%、从1%至20%、从1%至10%、从5%至10%、从10-20%、从10%至15%、从15%至20%、从1%至5%、从5%至15%;并且优选地含有至少六十种微量矿物质元素,浓度范围按重量百分比计从0.00001-3.0%,按重量百分比计从0.0001-1%、从0.001%至1%、从0.01%至3%、从0.1%至3.0%。微量矿物质元素包括铝、锑、砷、钡、铍、铋、硼、溴、镉、铈、铯、铬、钴、铜、镝、铒、铕、氟、钆、金、铪、钬、碘、铟、铱、铁、镧、铅、锂、镥、汞、钼,钕、镍、铌、钯、铂、镨、铼、铑、铷、钌、钐、钪、硒、银、锶、硫、钽、铽、碲、铊、钍、铥、锡、钛、钨、钒、镱、钇、锌和锆。
用于生产本发明的矿物质提取物组合物提取过程通常不将导入任何矿物质作为提取过程的一部分。因此,原材料,按提取方法处理的原始粘土或其它土壤,很可能包括与多种可探测矿物质一起自然富集的自然存在的硅酸铝和其它金属硅酸盐。如果矿物质元素在含水液体提取物中被鉴定和定量,通常,由于干燥过程或体积减小,该元素在干粉提取物中会被鉴定并定量成更高的浓度。
例如,使用美国专利申请系列号11/725,729(通过引用在此并入)所述的土壤和提取方法所生产的提取物组合物,使用鉴定和定量干燥形式和液体形式的矿物质提取物组合物的标准技术,通过进行化学分析的独立分析测试而检测。采用诸如滴定、电感耦合等离子体、质谱和原子吸收设备的科学上可接受的并且标准的设备,在Huntsville,Ala.的Teledyne Wah Chang实验室进行的测试结果产生含水矿物质提取物组合物和干燥矿物质提取物组合物的矿物质元素定量数据,列于表I中,所述干燥矿物质提取物组合物是当所述含水矿物质提取物组合物被干燥至粉末时产生。
表I:矿物质提取物组合物
列于上表I中的矿物质提取物组合物为从自然存在的土壤中产生,所述土壤的分析反映在下表II中。
表II:自然存在的土壤的分析
一旦得到所需要的自然存在的土壤或土壤组合物,土壤经受美国专利系列号11/725,729中所述的提取过程。在本发明的实施中,优选粘质土、粘质土混合物或粘质土与风化褐煤的混合物。优选该土壤组合物的一个原因在于该土壤可具有高含量的在本发明的实施中认定为重要的矿物质元素。如所述,优选根据本发明的矿物质提取物组合物包含至少八种常量矿物质元素和至少六十种微量矿物质元素。
确定粘质土是否可作为可接受的原始材料的第一步为确定砷、铅、汞和镉是否各自以可接受的低浓度存在。本发明的一个方面包括具有这些元素中的每一种的浓度的量比下表III中所示浓度低的组合物。
表III有毒元素的最高要求浓度
元素 | 要求的最高土壤浓度ppm或ppb |
砷 | 0.2ppm |
铅 | 0.17ppb |
汞 | 0.116ppm |
镉 | 1.12ppm |
表IV自然存在的土壤中所选的稀土元素的优选最低浓度
元素 | 优选的最低土壤浓度ppm |
铈 | 40 |
镨 | 2 |
钕 | 20 |
钐 | 3.5 |
铕 | 0.49 |
铽 | 0.62 |
镝 | 4 |
钬 | 1 |
铒 | 2 |
铥 | 0.65 |
镱 | 1.2 |
镥 | 0.45 |
列于表IV中的元素浓度可根据需要变化,但是应注意,需要所述元素至少具有表IV所示的每个元素的浓度。本发明的组合物的原始材料土壤可包含或不包含列于表IV的稀土元素之一或全部。例如,可在原始材料土壤中发现浓度为至少18ppm的镧和浓度为至少3.7ppm的钪。也可发现钷和钆的浓度。本发明的组合物的原始材料土壤可包含或不包含至少10种稀土元素,至少12种或更多的稀土元素以及可选择包括镧和钪。尽管不希望被局限于任何特定的理论,理论上,可以认为在土壤中和在衍生于原始材料土壤的矿物质提取物组合物中存在的稀土元素,当被身体摄入或透皮吸收时,用于提高矿物质提取物组合物的功效。
如美国专利申请系列号11/725,729所教导,提供或组合粘质土或粘土和土壤混合物以产生矿物质元素后,原始材料土壤就经受美国专利申请系列号11/725,729中所教导的提取过程以产生包含在本发明的抗糖化组合物中的矿物质提取物组合物。
通常,原始材料土壤的提取使用下列步骤。通常使用诸如逆向渗透的已知方法纯化的水在混合池中被加入到柠檬酸和原始材料土壤中。柠檬酸(或磷酸或其它可食用的酸)或其组合物的量可在所用水的重量的0.25%至7.5%的范围内,但是通常在1.0%至2.0%的范围内。水、柠檬酸和原始材料土壤形成泥浆,并且将其温和搅拌(例如,叶片以一至十RPM缓慢旋转)约一小时,尽管搅拌时间可根据需要变化。将池中的泥浆导向沉降池以使微粒向下沉出泥浆。泥浆在沉降池中保持任何需要的时间长度,在约一至十天的范围内。随着泥浆在沉降池中保持的时间长度的增加,可从池中抽出并输送至冷却池或浓缩器中的液体量增加,并且沉于池底的固体量增加。可使用添加剂以促进固体从泥浆中沉积。当泥浆停留在沉降池需要的一段时间后,将液体从池中抽至冷却池中,或直接至浓缩器中。池底的固体可被再处理、弃去或可以其他方式利用。
冷却池将来自沉降池中的液体冷却至40-70(5至21℃)的温度范围内。冷却的液体被输送到浓缩器。
浓缩器将水从冷却的液体中移除。这可使用诸如薄层复合逆向渗透系统或蒸发的已知方法完成。所产生的包含从原始泥浆中提取的矿物质的浓缩液体,按是否需要储存或进一步处理而注入冷却池或干燥器。冷却池将浓缩液体冷却至40至70(5至20℃)以防止酵母和霉菌生长。
由浓缩器产生的浓缩液体具有约为3的pH。浓缩液体通常包括按重量计从百分之三至百分之十二的矿物质元素,即如果矿物质元素从浓缩液体中分离,产生的干燥材料具有相当于浓缩液体重量的约3%至12%的重量。浓缩液体的pH通过改变添加至混合池的柠檬酸或其它可食用酸和/或碱性或酸性土壤的量来调节,并且在pH 2.0至pH 5.0的范围内,优选为pH 2.5至pH 3.5。浓缩液体(以及从其中产生的干燥粉末或其它材料)的pH优选为低于pH 4.5。可提供浓度至少为18%的矿物质提取物用于注射入干燥器。可利用任何所需的干燥系统,例如可喷入浓缩液体以产生粉末的塔。
本发明的抗糖化组合物可包括诸如表1中所示的矿物质提取物组合物,或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物,其与第二组分结合或混合。矿物质提取物组合物具有产生能量和胶原生长刺激的活性。矿物质提取物组合物的其它活性可在美国专利申请系列号10/725,729、11/472,536和11/638,311中发现,它们每个的全部内容在此并入。抗糖化组合物的第二组分可为液体组分或干燥组分,以形成液体组合物或干燥组合物。液体抗糖化组合物包括干燥粉末形式或液体形式的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物,和液体,所述液体包括但不限于水(可以是蒸馏水或碳酸水)和其它即用饮用或即用混合饮料,所述饮料包括但不限于咖啡、茶、能量饮料、果汁、牛奶和植物液体,诸如大豆制品、甘蔗制品、可可豆制品、蛋白质饮料、代餐饮料和诸如啤酒和红酒的含酒精制品。液体抗糖化组合物可包括与矿物质提取物组合物结合的药物组合物、营养组合物或食品补充组合物。例如,抗糖化组合物可包括与矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物结合的液体的药学上可接受的糖浆、赋形剂、填充剂或其它已知的药物制剂。作为进一步的实例,抗糖化组合物可包括在滴眼液的药物制剂中的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物以对例如糖尿病人的眼部提供抗糖化效应,以治疗视力障碍。
固体或干燥的抗糖化组合物包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物,以及固体或干燥材料,例如人类或动物摄入的食品。所述固体或干燥材料包括但不限于食物、食品、营养增甜剂和非营养增甜剂、药物组合物、营养组合物或食品补充组合物。例如,抗糖化组合物可以包括固体或干燥的药学上可接受的组合物、赋形剂、填充剂或其它已知的药物制剂,以与矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物结合制成诸如片剂、胶囊或粉末的剂量单位。本发明的组合物可作为食物的添加剂起作用,并且可与食品结合,所述食品包括其中可加入干燥或液体的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物以向食物提供抗糖化活性的食物。
作为特定的实例,抗糖化组合物可加入至人类或动物消耗的食品中。抗糖化增甜剂可包括单独的或与具有甜味或为公认的增甜剂的化合物或组合物相结合的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物。抗糖化组合物可用作增甜剂,并且可包括与增甜剂相结合的矿物质提取物组合物,所述增甜剂包括但不限于诸如蔗糖或枫糖浆的天然甜味化合物或组合物,或人工增甜剂。抗糖化矿物质提取物组合物可与包括罗汉果苷的增甜剂相结合,以形成抗糖化增甜剂罗汉果苷/矿物质提取物组合物。矿物质提取物组合物可被加入至增甜剂化合物或组合物中,并且以连续喷雾积聚至糖颗粒而用作例如包衣,或者加入诸如赤藓醇的可选的天然增甜剂以提供具有抗糖化性质的增甜剂。矿物质提取物组合物可作为液体加入至甜菊提取物以提供具有抗糖化性质的结合的甜菊/矿物质提取物组合物增甜剂。矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物可加入至已知的可提高AGE的增甜剂,诸如高果糖玉米糖浆,以提供抗糖化的矿物质提取物组合物、罗汉果苷/矿物质提取物组合物或罗汉果苷组合物/高果糖玉米糖浆组合物。
抗糖化活性或糖化抑制活性为可交换使用的术语,并且指化合物、提取物或组合物的能力,所述能力为干扰糖化反应以至于使生成修饰的氨基的蛋白质、脂类或核酸的氨基与糖的醛基或酮基的反应被改变或阻止,或者高级糖化终产物(AGE)的形成被体内或体外改变或阻止。
本发明的组合物包含自然增甜剂化合物,包括但并不局限于从罗汉果衍生而来的那些自然增甜剂化合物,其可单独或与诸如在此所教导的矿物质提取物组合物的组合物的其它组分相结合而用作抗糖化增甜剂。罗汉果为也公知为Momordica Grosvenori(罗汉果)或Siraitiagrosverorii(罗汉果)的植物,最初生长于中国并且现仍生长于中国以及气候允许的其它地方。通常,诸如公知为罗汉果苷的增甜剂化合物来源于植物罗汉果的果实,尽管该植物的任何部分都可被加工以产生增甜剂化合物。罗汉果果实为一种小的葫芦状果实,具有强烈甜味。所述水果包含称为罗汉果苷的自然存在的化合物,其被认为比蔗糖甜三百倍并且热量低。本文中,术语罗汉果苷用于指罗汉果果实中存在的一种或多种或者所有的罗汉果苷化合物,并且罗汉果苷也称为罗汉果提取物。在文献中,植物罗汉果也称为Lo Han Guo(罗汉果)或luohanguo(罗汉果),所有这些都意指同一种植物。
令人惊讶的是,发明者已经发现罗汉果苷具有抗糖化活性,并且具有能量提高和胶原生长刺激活性。本发明的方法包括将抗糖化活性、能量提高和胶原刺激的罗汉果苷组合物应用于甜味食品或饮料的组合物中。罗汉果苷组合物与诸如在此所教导的那些矿物质提取物组合物的组合在此称为罗汉果苷/矿物质提取物组合物。罗汉果苷组合物包括来自罗汉果果实的罗汉果苷提取物,并且制备罗汉果苷或罗汉果果实提取物的方法为公知的。
矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化活性的有效量可以依赖或可不依赖于矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物中所加入的第二组分的类型。抗糖化组合物可一天一次、一天两次、一天三次或一天更多次提供给或施用于受治疗者。抗糖化组合物可以固定方案或按所需的方案每天、每周或每月施用。通常,抗糖化组合物可含有从约0.0001g至约1000g矿物质提取物、或从约0.0001g至约100g、从约0.0001g至约10g、从约0.001g至约1000g、从约0.01g至约1000g、从约0.1g至约1000g、从约1.0g至约100g、从约1.0g至约10g、从约10g至约1000g,以及在其中的范围。例如,12oz增强的水饮料可含有0.1克矿物质提取物组合物,而含高果糖玉米糖浆的10oz碳酸饮料可含有10克矿物质提取物组合物,而且一加仑大小的枫糖浆可含有300克矿物质提取物组合物。
罗汉果苷/矿物质提取物组合物可含有约0.0001g至约1000g如上所述的矿物质提取物,该提取物以约0.0001g至约1000g的量加入到含有罗汉果苷的罗汉果苷组合物。
产生于原始材料土壤的选择和提取过程的矿物质提取物组合物包含在用于本发明的方法的抗糖化组合物中。本发明的方法包括抑制糖化终产物形成的方法、抑制AGE形成的方法、抑制蛋白质、脂质和/或核酸的糖化反应的方法、抑制与糖化反应相关的衰老效应的方法,以及治疗或预防糖化相关的症状的方法,或者防止糖化产物累积的方法,所述糖化相关的症状包括但不限于糖尿病(I型和II型)的并发症、类风湿性关节炎、阿耳茨海默氏病、尿毒症、神经毒性、动脉粥样硬化、炎性反应、心室肥大、血管病、心肌炎、肾炎、关节炎、肾小球性肾炎、微血管病和肾机能不全。抑制糖化或抑制AGE形成的方法、糖化相关症状的预防或治疗的方法包括施用或提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物于人类或动物,其中所述抗糖化组合物改变受治疗者的糖化效应、糖化活性、糖化速率或诸如AGE的糖化产物的量。例如,糖化活性的量通常可通过糖化血红蛋白的AIC测试来测量。AIC测量中的降低表明较低量的体内糖化活性。对糖化蛋白质的测量对于本领域技术人员是公知的,参见例如美国专利第5,506,144号。
本发明的方法包括制备和使用抗糖化组合物。该组合物可被健康的年轻或成年动物和人类,以及有风险发展为或患有糖尿病、动脉粥样硬化或相似的糖化相关症状的人类或动物所使用。食物、饮料和营养添加剂抗糖化组合物可用于向受治疗者提供抗糖化潜能,并且提供降低蛋白质和糖类的可能的交联以促进健康和健康状态的益处。例如,由于引入本发明的矿物质提取物组合物而具有抗糖化能力的食物或饮料系列因多种健康益处而被不同年龄的人使用。
治疗或预防糖化相关症状或抑制糖化水平的方法包括提供有效量的包括矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于肠炎。肠炎可影响免疫系统和神经系统,并且造成包括食物耐受不良、变态反应、炎症、认知困难、机能亢进和其它行为问题的许多症状。高级糖化终产物的受体(RAGE)为与炎性响应相关的细胞表面受体家族。目前认为,RAGE涉及当糖化的消耗促进产物与肠道和胃上皮衬细胞(cell lining)接触时肠道中的炎性响应。已经表明,RAGE在肠道上皮细胞中表达,主要集中于接近顶端细胞连接复合物的侧膜。尽管在通常情况下RAGE在上皮中表达低,但是在糖消耗时表达被上调。当食用糖时发生的糖化作用导致肠壁和胃壁的交联和炎症。
在预防或治疗糖化相关症状或抑制糖化产物和AGE的本发明的方法中,例如,包含诸如矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物的饮料或食物可被受治疗者(动物或人类)摄入以降低糖化对胃肠系统中衬细胞的影响。该抗糖化组合物可支持肠道抗糖化效应,提高肠道氧化化合物的去毒作用,提高肠道内营养吸收,帮助延迟肠炎进程,提高肠道作为AGE进入循环途径的看门者的作用,促进肠道健康和帮助降低炎症因子,以及帮助平静和缓解肠道糖化诱导的炎症。饮料和食物除了包含抗糖化组合物外,还可包含植物,诸如甘菊、蒲公英、紫锥花(echinecea)、大蓟(milk thistle)、龙胆、甘草、繁缕(chickenweed)、绣线菊、白毛茛、西班牙黑萝卜(Spanish black radish)和叶绿素。
预防或治疗糖化相关症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于认知功能。认知为覆盖脑功能的许多方面的通用术语,脑功能包括学习、记忆、思考和推理。这些过程在自然的衰老过程中或在变性疾病期间衰退。高级糖化终产物和自由基损伤可为衰老的自然部分,导致认知功能和运动能力的降低,并且可导致痴呆、阿耳茨海默病或帕金森病的加速形式。例如,包括抗糖化组合物饮料或食物可被受治疗者(动物或人类)摄入以降低糖化对神经系统的影响,降低炎性响应,并且防止循环系统的损伤。所述抗糖化组合物可用于避免认知退化,重建最优的认知功能,促进健康的认知功能,提高认知表现,促进健康的脑功能和帮助对抗氧化诱导的认知退化、加强认知功能防御,以及刺激相关的认知过程。饮料和食物除了包括抗糖化组合物外,还可包括以下的草药或植物化合物或提取物:银杏、人参、长春西汀(vipocetine)、绿茶、大豆异黄酮、维生素E、维生素C、维生素B6、维生素B12、磷脂(磷脂酰丝氨酸和磷脂酰胆碱)和胞磷胆碱(前体)和甘油磷酸胆碱、α-硫辛酸、乙酰左旋肉毒碱、辅酶Q10、肌酸、必需脂肪酸、DHA、EPA和白藜芦醇或葡萄籽提取物。
预防或治疗糖化相关的症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于视力。视力丧失、白内障和黄斑变性的主要的危险因子是眼部动脉、毛细血管和视网膜的硬化。已公认,糖尿病人比一般人群具有更高的视力障碍发病率,部分是因为高级糖化终产物在眼部积累。例如,包括抗糖化组合物的饮料或食物可被受治疗者(动物或人类)摄入,以降低糖化对眼部和相关结构的影响。该组合物用于提高视觉健康,防御衰老相关的视觉退化,重建眼部微循环,避免衰老相关的视力障碍,避免糖化引发的视力障碍,降低衰老相关的视力障碍,促进眼部卫生,提高晶状体透明度,并且保持健康视觉。饮料和食物除了包括抗糖化组合物外,还可包括草药,诸如越桔、银杏;植物营养素,诸如叶黄素、玉米黄素、番茄红素、生物类黄酮、混合胡萝卜素、硫辛酸、正乙酰半胱氨酸和槲皮素;氨基酸,诸如牛磺酸、谷胱甘肽、半胱氨酸;维生素,诸如维生素A、C、E、B-12、β-胡萝卜素;必需脂肪酸和其它抗氧化剂(也就是褪黑素)。
预防或治疗糖化相关的症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于关节炎。高级糖化终产物(AGE)被认为是如见于骨关节炎和类风湿性关节炎的炎症和最终的关节退化的促进因子。已经表明,AGE活化称为MMP的炎症调质以开始疼痛和受限的运动灵活性的级联效应。滑膜液被氧化并且经受糖化驱动的硬化。已鉴别,糖化标记在骨关节炎和类风湿性关节炎患者中以增高的量存在,表明糖化为这些症状的因素。本发明的抗糖化组合物在年幼者中的预防性使用可预防或延缓这些症状的发作,并且在任何年龄摄入可以降低骨关节炎和类风湿性关节炎的发病率和严重性。在任何年龄摄入,本发明的抗糖化组合物可以限制糖化的级联损害,并且因此降低促进伴随衰老的较长期的关节炎疾病的倾向。
例如,包含抗糖化组合物的饮料或食物或食品可被受治疗者(动物或人类)摄入以降低糖化对关节或滑膜液的影响。所述组合物可用于延缓衰老相关的关节炎性退化,保持关节和腱的灵活性,提高健康骨强度和关节弹性,促进骨结构完整性和灵活性,抵御糖化诱导的关节退化,提高关节和腱的健康滑膜环境,促进患关节炎时更积极的生活方式,促进针对关节炎的长期的预防性健康,以及降低的炎症、点肿胀和紧缩感的发病率。
饮料或食物,除了包括抗糖化组合物外,可还把包括草药和草药提取物,诸如姜、中国雷公藤、柳皮提取物、龙牙草,猫爪草、荨麻、乳香,S-腺苷甲硫氨酸(SAMe)、硫酸软骨素、葡萄糖胺、必需脂肪酸、以及诸如菠萝蛋白酶和槲皮素的酶。
预防或治疗糖化相关的症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于勃起机能障碍(ED)。已提出糖化通过形成蛋白质和DNA的加合物以及交联而在衰老相关的过程中起作用。这些交联可以通过清除勃起需要的一氧化氮而导致勃起机能障碍。另外,糖化导致动脉硬化,这抑制了保持勃起所必需的动脉灵活性。已证实,AGE通过影响海绵体的功能能力并通过干扰天然阴茎血管扩张剂、内皮和神经元一氧化氮(NO)的产生损害勃起功能。例如,包括抗糖化组合物的饮料或食物可被受治疗者(动物或人类)摄入以降低糖化对获得和维持勃起的能力的影响。所述组合物可用于提高健康的性功能,减缓衰老相关的ED循环退化,防止糖化诱导的ED,最大化性循环过程,提高健康的勃起性能,以及促进正常的勃起性能。饮料或食物除包括抗糖化组合物外,也可包括维生素C&E、生物类黄酮、必需脂肪酸、育亨树皮、淫羊藿、玛咖、沙巴棕和男宝(man bao)。
用于这些和其它糖化相关的症状的抗糖化组合物可包括即食谷物、果汁、糖果、口香糖、营养补充剂、风味水饮料、碳酸和非碳酸饮料、诸如啤酒和红酒的酒精饮料、婴儿食物以及许多其它食物和饮料。本发明的抗糖化组合物可用于动物饲料添加剂。
AGE被认为在降低细胞代谢速率和功能上起作用。本发明的抗糖化矿物质提取物组合物可在减缓由糖化终产物引起的代谢率降低中起作用。另外,本发明的抗糖化组合物也可通过提高线粒体功能而向体细胞提供能量。参见在此的实施例,其中在此教导的组合物在线粒体代谢中起作用。例如,包括抗糖化组合物的饮料或食物可被受治疗者(动物或人类)摄入以提高身体的能量代谢,所述抗糖化组合物包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物。所述组合物提供无化学刺激的代谢增强。
AGE被认为在降解胶原中起作用。本发明的抗糖化组合物可在减缓由于糖化终产物引起的胶原降解中起作用。预防或治疗糖化相关的症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于胶原生成和保持。胶原是组成人体总蛋白质量的25%的高度特征化的纤维蛋白家族。伴随衰老,胶原生成降低并且胶原降解增加。无论是否促进更年轻的皮肤,为了更健康的骨和腱,胶原刺激是食物和饮料成分所需的属性。I型胶原的刺激达到高于未处理(水)对照几乎90%的实施例显示于本文。基质金属蛋白酶MMP是消化和分解体内蛋白质的蛋白水解酶或蛋白酶。至今已发现超过30个不同类型的具有多种酶消化功能和炎症介导的MMP。对于皮肤,MMP酶的主要功能是保持皮肤基质,尤其是结构蛋白胶原再循环的稳定状态,并且一些皮肤基质为炎症途径的一部分。作为衰老的一部分,MMP倾向于过度表达其自身。另外,炎症、刺激和环境应激(自由基)提高皮肤的MMP水平以产生更多的胶原消化。在此教导的使用人类成纤维细胞的体外研究表明本发明的矿物质提取物组合物在浓度水平低至0.005%时有效地阻断胶原消化酶。在0.05%的浓度水平时,可见对MMP的100%的抑制作用。
AGE为自由基的氧化促进剂。本发明的抗糖化组合物可在抵抗由糖化终产物产生的自由基中起作用。本发明的方法包括治疗或抑制氧化,包括提供有效量的抗氧化剂组合物,该抗氧化剂组合物包含在此所述的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物与诸如食物或饮料的第二成分的组合。有许多环境和生活方式的因素可提高活细胞的氧化水平至不健康的水平,所有都加速衰老进程并且产生疾病的可能性。因为其对关键生物系统的损伤作用,氧化应激已经涉及超过100种疾病以及衰老。抗氧化剂紧密参与抑制细胞损伤——癌症、衰老和多种其它疾病的共同途径。耐力训练可提高氧气使用超过休息状态10至20倍。这大大提高自由基的产生,激发增加的对肌肉和其它组织的损伤的担心。饮料或食物,除包括抗氧化组合物之外,还可包括维生素A、C和E,以及植物多酚,所述氧化组合物包含本发明的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物。
尽管不希望局限于任何特定的理论,目前认为,皮肤衰老可被AGE和其对于HA和胶原的降解、自由基和炎症的作用而加速。如下面的实施例中所证实,公布于此的矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物可增加透明质酸(HA)的浓度。这也可辅助滑膜液环境。提高HA的能力用成年人类皮肤成纤维细胞以每孔10,000个细胞铺板于补充5%血清的高葡萄糖DMEM来评估,并且加入 矿物质提取物组合物。5天后,收集细胞培养物条件培养基,并且将每个试验条件下100ul的样品用于HA测试。HA测试使用来自Echelon(Salt Lake City,UT)的乙酰透明质酸酶联免疫吸附测试试剂盒(HA-ELISA,目录号K-1200)进行。观察到矿物质提取物组合物产生>20%的HA刺激。与抗糖化、胶原的刺激和保护、抗炎性性质和抗氧化性质联合,预防或治疗糖化相关的症状或抑制糖化产物和AGE的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物用于局部美容治疗或口服施用的美容产品,诸如抗皮肤衰老的饮料和食物。由于高级糖化终产物(AGE)已经被确认为炎症的促进剂,而炎症被确认为皮肤衰老的促进剂。
总得来说,本发明包括抗糖化组合物,以及制备和使用抗糖化组合物的方法,以及治疗和预防糖化相关症状的方法。治疗或防止糖化相关症状的方法,包括将有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物施用于人类或动物,其中所述量有效地减少至少一部分人类或动物的糖化事件。抗糖化组合物可包括矿物质提取物组合物,抗糖化组合物可包括罗汉果苷/矿物质提取物组合物,或者抗糖化组合物可包括罗汉果苷组合物。抗糖化组合物还可包括食物或饮料。糖化相关症状包括糖化终产物的形成,AGE形成,蛋白质、脂质和/或核酸的糖化反应,与糖化反应相关的衰老效应,以及糖尿病(I型和II型)的并发症、类风湿性关节炎、阿耳茨海默病、尿毒症、神经毒性、动脉粥样硬化、炎性反应、心室肥大、血管病、心肌炎、肾炎、关节炎、肾小球性肾炎、微血管病和肾机能不全,或糖化产物的积累。
抗糖化组合物可包括罗汉果苷和矿物质提取物组合物的组合,而且还可与食物或饮料结合。抗糖化组合物可包括罗汉果苷和矿物质提取物组合物的组合,而且还可与增甜剂组合物结合。
抑制糖化反应的方法包括提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物,并且抑制糖化反应。所述方法可包括包含矿物质提取物组合物的抗糖化组合物,包含罗汉果苷/矿物质提取物组合物的抗糖化组合物,或包含罗汉果苷组合物的抗糖化组合物。所述方法还可包括包含食物或饮料的抗糖化组合物。所述方法可包括向人类或动物体内提供抗糖化组合物或可体外提供。
在此所用定义
化学元素。由仅一种原子组成和由单个或组合组成的超过100种基本的金属和非金属物质中任何一种构成所有的物质,大多数这些在重量上轻于并且包括铀的物质发现于在自然界,而剩余物质通过引起原子核变化人工产生。
粘土。一种天然的或合成的胶质无光泽的泥土组合物,所述组合物包括大小小于约0.002毫米的铝和/或硅的微小片状分层颗粒,所述组合物在湿润时通常为可塑的,并且在自然产生时所述组合物包括分解的火成岩和/或变质岩。大多数粘土具有在约4.5至8.5的pH范围。天然或合成的粘土包括矿物质元素。除具有小于5微米的颗粒大小外,粘土可包括具有大于5微米大小的颗粒。
风化褐煤。一种软的,松散质地的具有低BTU值的煤。风化褐煤是腐殖酸盐,可含有按质量计多达70%的矿物质,可从褐煤中产生,可由于长期未被加热和施压至生成无烟煤、褐煤或烟煤所需的程度而自然产生,而且可包含堆肥作为组分。
矿物质。任何自然存在的化学元素或化合物。矿物质具有特征性的晶体结构和化学组成或组成范围。
矿物质元素。作为矿物质或在矿物质中的自然存在的化学元素。矿物质元素可通过合成或制造工艺生成,然而每种矿物质元素确实作为矿物质或在矿物质中自然存在。
稀土或稀土元素。具有原子序数58至71的一族金属元素中的任何一个,包括铈、镨、钕、钷、钐、铕、钆、铽、镝、钬、铒、铥、镱和镥。在自然中,稀土元素与非金属元素以磷酸盐、碳酸盐、氟化物、硅酸盐和钽酸盐的形式结合。
沙。一种由小但是容易区别的颗粒组成的松散材料,所述颗粒直径通常小于2毫米并且大于约0.02毫米,最常见为岩石破碎产生的石英。
泥浆。未固化的或松散的沉积材料,其组成的岩石颗粒为比沙粒细并且比粘土颗粒大,特别是由直径范围从约0.02毫米至0.002毫米的矿物质土壤颗粒组成的材料。
必须指出,如在本说明书以及附加权利要求中使用,除非上下文清楚地另外指出,否则单数形式“一(a)”、“一(an)”和“所述(the)”包括复数指示物。
在此包括的所有专利、专利申请和参考文献的全部内容通过引用在此具体并入。
当然,应理解,前述仅指本发明的优选的实施方式,并且其中可对本发明进行多种改良和改变而没有背离本公开中所列的本发明的精神和范围。
本发明进一步通过以下实施例阐释,所述实施例不应被理解为以任何方式对其范围施加限制。相反,可以清楚理解的是,本领域内专业人员可在阅读此说明后,借助其进行多种的其它实施方式、改良及其等同物,且不背离本发明的精神和/或所附权利要求的范围。
实施例
实施例1
评价三种矿物质提取物组合物以测试抗糖化效应。
每个反应混合物含有10mg/ml在PBS中的白蛋白(Sigma)和500mM在PBS中的葡萄糖(Sigma G8270)。阴性对照为10mg/ml白蛋白,不含葡萄糖。阴性对照用于每个实验点。阳性对照为10mg/ml白蛋白,含有500mM葡萄糖和1mM盐酸氨基胍(Sigma 396494)。
样品制备
在I型无菌水中稀释每个矿物质提取物组合物至10mg/ml,通过0.22微米过滤除菌,并且在37℃下在100ug/ml、10ug/ml、1ug/ml和0.1ug/ml的稀释液中,在叠氮化纳存在下,在含有或不含有葡萄糖时,与反应混合物一起孵化11天。
如所述(Argirova和Argirov,2003),使用微板荧光仪Cytofluor2350(Millipore)通过非色氨酸荧光(激发于360nm,发射于460nm)的增加检测蛋白质糖化。通过减去背景读数(不含葡萄糖的样品)而得到每一个实验点的糖化值。
结果与讨论
如图1所见,0为阴性对照,并且每个矿物质提取物组合物显示出在50ug/ml下抑制白蛋白糖化,且#1显示总体最佳抑制活性。氨基胍(AG)(1mM)具有良好的抑制活性,此为其已知活性。
实施例2
抗糖化活性
正常人类皮肤成纤维细胞(NHDF)在允许大量细胞外基质(ECM)合成的条件下培养。NHDF以充分形式在正常培养基中生长至汇聚。在培养基中填加维生素C以诱导基质合成/沉积。
用三种浓度的矿物质提取物处理细胞(或不处理阴性对照)或用阳性对照的参照化合物(氨基胍)处理细胞。在所有条件下重复三次进行。
在细胞中加入过量的葡萄糖,并且在37℃孵化细胞15天。最后,提取并纯化ECM蛋白质(主要是胶原),并且将每个样品的一部分以可重现的点上样于硝酸纤维素。
使用抗-AGE抗体显示AGE导入到胶原中,并且用过氧化酶结合物和化学发光反应(ECL)对其标记。使用化学发光成像仪进行相对信号鉴定。
结果列于如下表1和表2中。
表1
第一个膜 | |||||
处理 | 浓度 | 强度(AU) | 平均值 | Sem | %对照 |
对照 | - | 49908 | 53561 | 1843 | 100 |
54965 | |||||
55809 | |||||
氨基胍 | 1mg/ml | 24719 | 33454 | 4377 | 62 |
38328 | |||||
37316 | |||||
矿物质提取物组合物1 | 0.1μg/ml | 46270 | 42886 | 2991 | 80 |
45465 | |||||
36922 | |||||
25μg/ml | 48675 | 51128 | 6560 | 95 | |
63517 | |||||
41193 | |||||
250μg/ml | 38893 | 44343 | 2755 | 83 | |
47772 | |||||
46363 |
矿物质提取组合物2 | 0.1μg/ml | 48856 | 48340 | 1823 | 90 |
51209 | |||||
44956 | |||||
25μg/ml | 51233 | 44177 | 3658 | 82 | |
38977 | |||||
42322 | |||||
250μg/ml | 43883 | 40454 | 3144 | 76 | |
43304 | |||||
34176 | |||||
矿物质提取组合物3 | 0.1μg/ml | 40338 | 45520 | 3833 | 85 |
53004 | |||||
43219 | |||||
25μg/ml | 46324 | 45936 | 1593 | 86 | |
48480 | |||||
43003 | |||||
250μg/ml | 37197 | 41239 | 2569 | 77 | |
46008 | |||||
40513 | |||||
矿物质提取组合物4 | 0.1μg/ml | 34161 | 42089 | 4034 | 79 |
47349 | |||||
44758 | |||||
25μg/ml | 43224 | 50402 | 7095 | 94 |
64592 | |||||
43389 | |||||
250μg/ml | 44930 | 42543 | 4084 | 79 | |
48115 | |||||
34585 |
sem:平均值的标准误差
AU:任意单位
表2
膜2 | |||||
处理 | 浓度 | 强度(AU) | 平均值 | Sem | %对照 |
对照 | - | 53903 | 60294 | 4067 | 100 |
59133 | |||||
67846 | |||||
氨基胍 | 1mg/ml | 36460 | 38752 | 1414 | 64 |
38464 | |||||
41333 | |||||
膜2 | |||||
处理 | 浓度 | 强度(AU) | 平均值 | Sem | %对照 |
对照 | - | 53903 | 60294 | 4067 | 100 |
59133 | |||||
67846 | |||||
氨基胍 | 1mg/ml | 36460 | 38752 | 1414 | 64 |
38464 |
41333 | |||||
矿物质提取物组合物5 | 0.1μg/ml | 38940 | 45072 | 3066 | 75 |
48090 | |||||
48186 | |||||
25μg/ml | 52306 | 56723 | 2224 | 94 | |
58468 | |||||
59394 | |||||
250μg/ml | 50460 | 51318 | 439 | 85 | |
51585 | |||||
51908 | |||||
矿物质提取物组合物6 | 50μg/ml | 40393 | 46127 | 3053 | 77 |
47176 | |||||
50811 | |||||
200μg/ml | 36099 | 45666 | 5859 | 76 | |
44590 | |||||
56308 | |||||
500μg/ml | 44656 | 50828 | 3314 | 84 | |
51825 | |||||
56004 |
sem:平均值的标准误差
AU:任意单位
矿物质提取物组合物显示降低糖化终产物(AGE)的生成的倾向。
实施例3
本测定的目的为使用MTT测定测试在细胞培养群落中矿物质提取物组合物(6批次)对线粒体代谢的作用。MTT测定测量琥珀酸脱氢酶的活性,该酶为线粒体中呼吸电子传递链的关键酶(Berridge & Tan,1993)。
方法
在I型无菌水中稀释矿物质提取物组合物至10mg/ml,通过0.22微米过滤除菌,并且以500ug/ml、50ug/ml、5ug/ml和0.5ug/ml的稀释液,与每孔2,500个来自81岁女性捐献者(Cascade Biologies,批号061215-901)的人类皮肤成纤维细胞,在含有2.5%小牛血清的高葡萄糖DMEM培养基中孵化72h。
对于线粒体代谢的测量,在实验结束时,在细胞培养物中加入MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑,Sigma目录号M5655)并且孵化2h。然后弃去细胞培养基并且将细胞内MTT还原产物甲月替溶解于100%DMSO中。与细胞培养物中线粒体活性成比例的比色信号用BioRad微板分光光度计3550-UV在570nm下测量。
对于细胞数的测量,在实验结束时,使用磺酰罗丹明B染色细胞骨架蛋白,并且用BioRad微板分光光度计3550-UV在570nm下测量与细胞数成比例的比色信号。
通过确定两个相关的信号的比,标准化线粒体代谢的细胞数。
结果与讨论
参见图2A和B的结果。所有测试的矿物质提取物组合物批次显示相似的模式,且一个批次(#2)可刺激线粒体代谢高达基线上几乎100%。参见图2A。0为阴性对照,其中将水加入细胞中,且六个矿物质提取物组合物样品中每一个标记为1-6,6个样品中每一个浓度为500ug/ml、50ug/ml和5ug/ml和0.5ug/ml。C为肌酸,以500ug/ml加入细胞。
有趣地是,来自年纪较大的捐献者的皮肤成纤维细胞对肌酸(6.3%刺激)仅轻微敏感,并且对碱性成纤维细胞生长因子(bFGF,结果未显示)不敏感。此外,重要地是,矿物质提取物组合物在此测定中不刺激细胞增殖,仅刺激代谢。参见图2B。
实施例4
皮肤成纤维细胞分泌的胶原为皮肤细胞外基质的主要组分。在衰老的和光损伤的皮肤里,新的胶原池由于皮肤成纤维细胞的较差的量和质量而降低。本方案的目的是测试在人类皮肤成纤维细胞条件培养基中矿物质提取物组合物对I型胶原的作用。
方法
将正常人类皮肤成纤维细胞(第7代,批号7F1254,Cambrex,Walkersville,MD)在96孔板中(板#431)中接种至含有5%胎牛血清的DMEM培养基中,并且使其生长至后期分会合阶段。在I型无菌水中制备矿物质提取物组合物的水溶液,并且以1/20稀释加入细胞培养物中。水为未处理对照,并且抗坏血酸磷酸镁(MAP目录号A8960,Sigma,St.Louis,MO)为阳性对照。在实验开始5天后收获细胞培养条件培养物,并且根据标准ELISA程序(Dobak等人,1994,Zhao等人,2005),使用亲和纯化的抗体通过夹心ELISA,之后通过抗生蛋白链菌素-卵白素-HRP结合物和ABTS测定I型胶原。
在405nm下用BioRad微板分光光度计3550-UV测量与胶原含量成比例的比色信号。参见图3,矿物质提取物组合物在HDF-条件培养基中对I型胶原的作用。
结果与讨论
对胶原I的强刺激见于阳性对照(MAP),参见图3。矿物质提取物组合物显示对I型胶原极好的、钟形的刺激,高达超过非处理(水)对照的几乎90%,与阳性对照(MAP)相当。钟形的剂量反应曲线表明矿物质提取物组合物对I型胶原的刺激的最优浓度为在0.01和0.1mg/ml(0.1-1%)之间。
实施例5
金属蛋白酶(MP)(胶原蛋白酶)活性用来自Molecular Probes(Invitrogen,IL)的Enzcheck试剂盒使用猝灭的荧光明胶和种属金属蛋白酶梭菌(Clostridium)胶原蛋白酶IV测量。500ug/ml、50ug/ml和5ug/ml和0.5ug/ml的稀释液中的矿物质提取物组合物,在胶原蛋白酶底物-猝灭的荧光素联明胶存在下并且在蛋白水解酶胶原蛋白酶存在下孵化。100ug/ml的有效的MP抑制剂菲咯啉用作阳性对照。使用Millipore Cytofluor 2350显微荧光计在激发/发射波长为485/530nm下测量释放被消化的荧光明胶的动力学。
结果与讨论
如图4所示,矿物质提取物组合物具有金属蛋白酶抑制剂活性。该活性可在浓度低至50ug/ml(6%抑制)下被检测,并且在500ug/ml时为100%抑制。如所预期的,MP活性完全被菲咯啉所抑制。
多数金属蛋白酶抑制剂为无差别的二价阳离子螯合剂。矿物质提取物组合物可通过不同的作用机理起作用。值得研究地是,这种可能的新颖的作用机理是否可削弱对金属蛋白酶亚型的特异性,这是皮肤病学应用所关注的。
实施例6
使用标准荧光测定技术,在独立的体外实验中测定矿物质提取物组合物的抗氧化效应以评价氧自由基吸收能力(ORAC分数)。
已经报道了主要的抗氧化水果具有在15-30范围内的ORAC分数,这是极其显著的抗氧化能力。使用同样的程序测试,发现矿物质提取物组合物样品在相等的重量基础上,ORAC分数超过主要的抗氧化水果10-30倍之间。
水果的ORAC(μmole TE/g)
樱桃 15
草莓 24
覆盆子 28
黑莓 28
蓝莓 28
石榴 32
矿物质提取物组合物的氧自由基吸收能力
因为氧化应激对重要的生物系统的损伤作用,氧化应激已经涉及超过100种疾病以及衰老(Ames等人,1993)。本测试的目的是应用氧自由基吸收能力(ORAC)测定测量若干矿物质提取物组合物样品的抗氧化潜能。
方法
根据描述于Ou等人(2001)并少量修改(Sunny BioDiscovery程序#21)的方法进行ORAC测定。ORAC测定测量抗氧剂组分抑制荧光素二钠(FL)(Sigma-Aldrich,St Louis,Mo)荧光降低的能力,这种降低由过氧化自由基发生剂,2′,2′-偶氮双(2-脒基丙烷)二盐酸(AAPH)(Wako Chemicals,Richmond,Va)诱导。
矿物质提取物组合物样品在I型无菌水中稀释至10mg/ml,通过0.22微米过滤来除菌,并且10ug/ml加入反应混合物中。
浓度为1ug/ml、5ug/ml和10ug/ml的Trolox(在I型水中新鲜配制)被用于生成标准曲线,其通过绘制用出NIH ImageJ软件计算的曲线下面积(AUC)对表示为微摩尔/升的Trolox浓度的曲线。然后每个样品的AUC用于Trolox标准曲线。参见表3。
表3ORAC值
矿物质提取物组合物(样品号) | 抗氧化活性(umoles TE/g) |
1 | 1000 |
2 | 925 |
3 | 1400 |
4 | 925 |
实施例7抗糖化组合物
抗糖化组合物包括
新鲜橙汁98.80%(重量百分比)
山梨酸钾0.15%
维生素E(生育酚)0.05%
表Ⅰ中干燥的矿物质提取物组合物1.00%
过程:
1.使用合适的压榨装置,挤压橙汁
2.加入和混合于山梨酸钾和生育酚
3.加入和混合矿物质提取物组合物
4.在5-8摄氏度包装和冷却
一升加工的橙汁中的矿物质含有不低于1ppm的常量矿物质,所述常量矿物质由钙、氯、镁、锰、磷、钾、硅、钠的混合物组成以及不低于0.0001ppm的微量矿物质,所述微量矿物质由以下组成:铝、锑、砷、钡、铍、铋、硼、溴、镉、铈、铯、铬、钴、铜、镝、铒、铕、氟、钆、金、铪、钬、碘、铟、铱、铁、镧、铅、锂、镥、汞、钼,钕、镍、铌、钯、铂、镨、铼、铑、铷、钌、钐、钪、硒、银、锶、硫、钽、铽、碲、铊、钍、铥、锡、钛、钨、钒、镱、钇、锌、锆的混合物。
实施例8
饮料添加剂-粉末状浓缩矿物质包装
将7g表I中干燥的矿物质提取物组合物包装于箔包装中。将箔包装中的7g干燥矿物质提取物组合物混合至12盎斯包括水的任何饮料,递送不低于1ppm的常量矿物质以及不低于0.0001ppm的微量矿物质,所述常量矿物质由钙、氯、镁、锰、磷、钾、硅、钠的混合物组成,所述微量矿物质由以下组成:铝、锑、砷、钡、铍、铋、硼、溴、镉、铈、铯、铬、钴、铜、镝、铒、铕、氟、钆、金、铪、钬、碘、铟、铱、铁、镧、铅、锂、镥、汞、钼,钕、镍、铌、钯、铂、镨、铼、铑、铷、钌、钐、钪、硒、银、锶、硫、钽、铽、碲、铊、钍、铥、锡、钛、钨、钒、镱、钇、锌、锆的混合物。
实施例9
罗汉果苷/矿物质提取物组合物及其组分对1型胶原的作用
通过将约84%w/w%的3%液体矿物质提取物组合物与柠檬酸(0.01%w/w%)和防腐剂(至少0.01%w/w%)和来自酪蛋白水解产物的二肽及三肽(1.0%w/w%)结合制备罗汉果苷/矿物质提取物组合物。该混合物与可溶性食物纤维(1.50%w/w%)以及改良的食物淀粉(1.60%w/w%)结合。该混合物与山梨醇溶液,70%USP/FCC(9.0%w/w%)以及粉末状的罗汉果苷(罗汉果水果提取物)(MB North America)(3.0%w/w%)混合。该混合物与调味剂(0.5%w/w%)结合。在此称为SS II。
胶原是结缔组织(筋膜)、软骨、韧带、腱、骨和牙齿的主要组分,并且其还向血管提供结构支持。此测定的目的为确定罗汉果苷/矿物质提取物组合物及其单个组分对在人类皮肤成纤维细胞条件培养基中的I型胶原水平的作用。
方法
在所述测定中检测批号100-151106的矿物质提取物组合物(TL)的~3%溶液,和在此所教导的罗汉果苷/矿物质提取物组合物(SS II),罗汉果提取物中的罗汉果苷(Luo HG,批号MOGO 1-060502),赤藓醇(Eryt,批号07241BP952)和木糖醇(Xylit,批号H125T7G1)。所有测试材料保存于室温下。材料在1型水中稀释,通过0.22μ滤器过滤并且在TL和SS II最终浓度为1%、0.5%、0.1%、0.05%和0.01%(v∶v)以及Luo HG、Eryt和Xylit最终浓度为0.5%、0.1%、0.05%和0.01%(w∶v)下,在含5%小牛血清的高葡萄糖DMEM中的成年人类皮肤成纤维细胞(HDF,捐献者年龄为58岁,Lonza,批号7F39=019胸部)上测定。水为未处理对照并且抗坏血酸磷酸镁(MAP目录号A8960,Sigma,St.Louis,MO)为阳性对照。在实验开始72h后,收获细胞培养条件培养基,并且根据标准ELISA程序(Dobak等人,1994,Zhao等人,2005),通过夹心ELISA使用亲和纯化抗体,之后为链酶抗生物素-抗生物素-HRP结合物和ABTS测定细胞培养条件培养基的I型胶原。在405nm下用BioRad微板分光光度计3550-UV测量与胶原含量成比例的比色信号。
结果与讨论
如图5所示,TL和SS II在1%和0.5%下具有强的剂量依赖胶原刺激活性(TL高达54%以及SS II高达49%)。Luo HG在0.1%和0.05%下具有强力的剂量依赖刺激活性(高达77%)。相反,赤藓醇和木糖醇在所有被测浓度下不具有显著活性。0为水,未处理对照,使用50μg/mL MAP,Luo HG在图5中以L标记。
实施例10
罗汉果苷/矿物质提取物组合物、矿物质提取物组合物、三氯半乳蔗糖和赤藓醇对线粒体代谢的作用
该实施例的目的为使用MTT测定测试4个样品的作用:罗汉果苷/矿物质提取物组合物(在图6A中标记为1)、矿物质提取物组合物(在图6A中标记为2)、三氯半乳蔗糖浓缩液(在图6A中标记为3)和赤藓醇(在图6A中标记为4)对线粒体代谢I成纤维细胞群体的作用。MTT测定检测琥珀酸脱氢酶的活性,琥珀酸脱氢酶是线粒体(mitrochondria)呼吸电子传递链中的关键酶(Berridge & Tan,1993)。
方法
测试材料为如上实施例9中制备的罗汉果苷/矿物质提取物组合物,矿物质提取物组合物(~3%溶液)、三氯半乳蔗糖浓缩液(按重量计25%三氯半乳蔗糖)-批号F2304B311LB和赤藓醇(25%水溶液)-批号RFA 2104-35,并且在室温下保存。材料在1型无菌水中稀释,通过0.22μ滤器过滤并且在最终浓度为2.5%、0.5%和0.05%(v∶v)下,在含5%小牛血清的高葡萄糖DMEM中的人类皮肤成纤维细胞(Invitrogen)上检测72h。
为了在整个细胞群体中测量线粒体代谢,在实验结束时在细胞培养物中加入MTT(3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑,Sigma目录号M 5655)并且继续孵化另外的2小时。然后弃去细胞培养基,并且将细胞内MTT还原产物甲月替溶解于90%异丙醇中。细胞培养物中与线粒体活性成比例的比色信号用BioRad微板分光光度计3550-UV在570nm下测量。
结果与讨论
结果表明罗汉果苷/矿物质提取物组合物具有代谢刺激活性。矿物质提取物组合物具有剂量依赖活性,而在测试浓度下,三氯半乳蔗糖抑制细胞代谢并且赤藓醇不具活性(参见图6A)。通过重复每个实验材料的一个剂量来进一步证实并再现该第一个实验(重复四次进行)的结果(参见图6B)。碱性成纤维细胞生长因子(FGF在15nM下)用作阳性对照并且如所预见,显示出代谢刺激。0代表水,未处理的样品。
实施例11
罗汉果苷/矿物质提取物组合物及其组分对蛋白质糖化的作用
Maillard反应(非酶糖化)为产生诸如皮肤上细胞外基质蛋白质的大分子的胺基基团与还原糖上的羰基基团的交联的一系列反应。这些交联的大分子称为高级糖化终产物(AGE)。AGE的生成被自由基提高。AGE为诸如糖尿病和动脉粥样硬化的生理病理学标记,并且与衰老和光衰老过程相关联(van Boekel等人,1991)。此测定的目的为确定罗汉果苷/矿物质提取物组合物及其单个组分对白蛋白/葡萄糖模型系统中AGE形成的作用。
方法
矿物质提取物组合物(TL,-3%溶液)和罗汉果苷/矿物质提取物组合物(SS II)、罗汉果提取物(L,批号MOGO 01-060502)、赤藓醇(E,批号07J241BP952)和木糖醇(X,批号H125T7G1)保存于室温下。材料在1型无菌水中稀释,通过0.22μ滤器过滤并且在37℃下,TL和SS II最终浓度1%(v∶v)和L、E和X最终浓度为0.5%、0.1%、0.05%和0.01%(w∶v)下,在存在或不存在葡萄糖(Sigma G8270)下,在含有10mg/ml白蛋白和叠氮化纳的培养基中检测3周。抑制剂阳性对照为1mM盐酸氨基胍(AG)(Sigma 396494)。
如前所述(Argirova和Argirov,2003),使用微板荧光仪Cytofluor 2350(Millipore)通过非色氨酸荧光(激发于360nm,发射于460nm)的增加检测蛋白质糖化。通过减去背景读数(不含葡萄糖的样品)得到每一个实验点的糖化值并且表示为对照(水)的%。
结果与讨论
结果表明矿物质提取物组合物和罗汉果苷/矿物质提取物组合物在浓度为1%时具有中等的(20-25%)糖化抑制活性,而且在测试的所有其它浓度下无抑制活性。在此模型系统中,罗汉果提取物在所有测试的浓度下显示出剂量依赖性糖化抑制作用(在0.5%时可达几乎90%抑制)。在较低浓度下,木糖醇没有作用,并且赤藓醇具有中等的抑制作用。
氨基胍(AG)显示强的糖化抑制活性,表明本实验的技术成功。参见图7。
实施例12
本实验的目的是确定矿物质提取物组合物在0.5%(v∶v)时对成年人成纤维细胞的基因表达模式的作用
方法
人类成纤维细胞(批号7F3019,胸部,捐献者为58岁)从Lonza,Inc.获得,并且根据制造商的说明书培养。将0.5%(v∶v)的矿物质提取物组合物(批号100-151106)加入至在75cm2烧瓶中的在DMEM(高葡萄糖)/5%FCS中的汇合的确立细胞培养物,持续48小时。孵化完成时,细胞用胰蛋白酶处理,沉淀并在液氮中速冻。沉淀含有约4百万个细胞。
然后用Qiagen试剂盒对细胞进行RNA提取。提取的RNA质量通过电泳测定两次(提取之后和微阵列分析之前)。使用来自PhalanxBiotech(Palo Alto,CA)的人类OneArray平台将样品技术性复制地杂交。然后,进一步内部处理在超过30,000个探针上提取信息的Excel文件以排除具有高p值(p>0.1)和低倍变化(<T)的差异。所有描述为“未归类的”和“假定的”基因从最终分析中排除。使用ArrayStudio V2.5(Omicsoft)软件以确定受测试材料影响的功能类别。
结果与讨论
分离的RNA的数量和质量以及核酸与蛋白质的比极佳,表明RNA不仅是完整的而且不含蛋白质杂质。微阵列杂交是成功的。用矿物质提取物组合物处理显示基因表达模式的变化。矿物质提取物组合物调节2914个探针的表达,代表约9%的总探针。Array Studio分析显示矿物质提取物组合物显著地调节20个基因功能类别。
细胞代谢类别被最显著地调节。例如,观察到线粒体核糖体蛋白的前体和NADH脱氢酶的一致的上调,表明线粒体代谢通过涉及ATP生成的效应物的合成的上调而活化。提高的线粒体活性的进一步的指征是细胞色素C氧化酶的上调,细胞色素C氧化酶是线粒体呼吸链末端酶,该酶在调节能量的有氧生成上起关键作用。
另一个显著调节的类别是涉及鞘脂代谢过程的类别。可见超过5倍对MD-I蛋白质的表达。所述基因/蛋白质涉及多糖鞘糖脂的合成,多糖鞘糖脂是涉及细胞膜完整性的分子。有利于此途径的化合物可用于皮肤护理的应用。被上调的其它基因包括涉及解毒/抗氧化活性(对过氧化物氧还蛋白-5和超氧化物歧化酶-SOD-表达的刺激超过2倍)、细胞死亡抑制(通过半胱天冬酶途径的抑制)以及透明质酸和I型胶原合成的上调的那些基因。
实施例13
干燥增甜剂组合物
组合物为包含矿物质提取物组合物、罗汉果苷的罗汉果提取物和其它成分以形成增甜剂的增甜剂的实例。参见下表中的具体量。
成分 | C.T.F.A.名称 | W/W% |
A相 | --A相 | A相 |
1-矿物质提取物粉末 | 1-矿物质提取物粉末 | 6.00 |
2-无水柠檬酸,USP,FCC | 2-无水柠檬酸,USP,FCC | 0.20 |
3-苯甲酸钠,NF,FCC | 3-苯甲酸钠,NF,FCC | 0.20 |
4-山梨酸钾,NF,FCC | 4-山梨酸钾,NF,FCC | 0.20 |
5-Nutriose FM 06 | 5-糊精(可溶性食物纤维) | 3.50 |
6-N-Zorbit M | 6-木薯麦芽糖糊精 | 52.90 |
7-Sorbogem 712结晶山梨醇NF/FCC | 7-Sorbitot,NF/FCC | 10.50 |
8-罗汉果果实浓缩物 | 8-罗汉果(MomordicaGrosvenori Swingle) | 5.00 |
9-Aerosil 200Pharma | 9-硅 | 1.00 |
B相 | B相 | B相 |
9-N-Zorbit M | 9-木薯麦芽糖糊精 | 20.00 |
10-调味剂 | 10-调味剂 | 0.50 |
总数 | 100.00 |
另外的成分或代替物可从以下序列中选择。
1-食物淀粉改良剂、膨胀剂和/或功能剂:
·木薯麦芽糊精.
·小麦糊精
·糯玉米
·木薯淀粉
··成膜剂
·固化剂
·调味剂载体
·胶凝剂
·基于粮食的食物成分
·水解的谷物固体
·麦芽糖糊精,也参见膨胀剂;包封成分
·肉类膨胀剂
·油、植物、玉米
·改良的、预胶化的稀糊的淀粉、玉米、撒粉
·制片剂
·标准稳定胶质,如:琼脂、藻酸盐、阿拉伯树胶、角叉菜胶、羧甲基纤维素(CMC)、羟丙基甲基纤维素、微晶纤维素和甲基纤维素、刺槐豆胶、瓜尔胶(植物瓜尔豆(Cyamopsis tetragonoloba)的胚乳)、菊粉(可从菊苣根和耶路撒冷菊芋(Jerusalem artichokes)中提取的现有的食物纤维)、果胶(果胶从柑橘属水果皮中提取)、黄原胶、阿拉伯树胶。
营养制品成分:包括可在人类和/或动物的食物和健康之间建立联系的任何自然的和/或合成的类型。营养制品成分可包括从水果、草药、海产、香料、蔬菜和合成产生的化合物中取得的成分。这些种类包括但不限于下列成分:异黄酮、卵磷脂、油和脂肪(低级反式脂肪油)、植物甾醇、大豆蛋白、维生素、维生素E和混合生育酚。
调味剂:任何自然的和/或合成的调味剂可引入所述组合物中。可使用调味剂以促进组合物的使用。
Claims (20)
1.一种治疗或预防糖化相关症状的方法,包括
向人类或动物施用有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物,其中所述量有效地降低人类或动物中的至少部分的糖化事件。
2.如权利要求1所述的方法,其中所述组合物包括矿物质提取物组合物。
3.如权利要求1所述的方法,其中所述组合物包括罗汉果苷/矿物质提取物组合物。
4.如权利要求1所述的方法,其中所述组合物包括罗汉果苷组合物。
5.如权利要求1所述的方法,其中所述组合物还包括食物。
6.如权利要求1所述的方法,其中所述组合物还包括饮料。
7.如权利要求1所述的方法,其中所述糖化相关症状包括糖化终产物的形成、AGE形成,蛋白质、脂质和/或核酸的糖化反应、与糖化反应相关的衰老效应,以及糖尿病(I型和II型)的并发症、类风湿性关节炎、阿耳茨海默病、尿毒症、神经毒性、动脉粥样硬化、炎性反应、心室肥大、血管病、心肌炎、肾炎、关节炎、肾小球性肾炎、微血管病和肾机能不全,或糖化产物的累积。
8.一种抗糖化组合物,包括罗汉果苷和矿物质提取物组合物。
9.如权利要求8所述的组合物,还包括食物或饮料。
10.如权利要求8所述的组合物,还包括增甜剂组合物。
11.如权利要求8所述的抗糖化组合物,其中所述组合物包括罗汉果苷,并且用于能量产生。
12.如权利要求8所述的抗糖化组合物,其中所述组合物包括矿物质提取物组合物,其中所述组合物用于能量产生。
13.一种抑制糖化反应的方法,包括
a)提供有效量的包含矿物质提取物组合物或罗汉果苷/矿物质提取物组合物或罗汉果苷组合物的抗糖化组合物,且
b)抑制糖化反应。
14.如权利要求13所述的方法,其中所述组合物包括矿物质提取物组合物。
15.如权利要求13所述的方法,其中所述组合物包括罗汉果苷/矿物质提取物组合物。
16.如权利要求13所述的方法,其中所述组合物包括罗汉果苷组合物。
17.如权利要求13所述的方法,其中所述组合物还包括食物。
18.如权利要求13所述的方法,其中所述组合物还包括饮料。
19.如权利要求13所述的方法,其中所述组合物体内提供于人类或动物。
20.如权利要求13所述的方法,其中所述组合物被体外提供。
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PCT/US2008/079632 WO2009049246A1 (en) | 2007-10-10 | 2008-10-10 | Anti-glycation methods and compositions |
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EP (1) | EP2207420A4 (zh) |
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EP2207420A1 (en) | 2010-07-21 |
CA2702339A1 (en) | 2009-04-16 |
US20090226545A1 (en) | 2009-09-10 |
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US9107869B2 (en) | 2015-08-18 |
US9504713B2 (en) | 2016-11-29 |
EP2207420A4 (en) | 2011-01-12 |
WO2009049246A1 (en) | 2009-04-16 |
US10016450B2 (en) | 2018-07-10 |
US20150150909A1 (en) | 2015-06-04 |
US20170136050A1 (en) | 2017-05-18 |
US8927031B2 (en) | 2015-01-06 |
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