CN113769007A - 包含植物萃取物的组合物及其用途 - Google Patents
包含植物萃取物的组合物及其用途 Download PDFInfo
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- CN113769007A CN113769007A CN202111031378.4A CN202111031378A CN113769007A CN 113769007 A CN113769007 A CN 113769007A CN 202111031378 A CN202111031378 A CN 202111031378A CN 113769007 A CN113769007 A CN 113769007A
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Abstract
一种用于减少脂肪生成细胞内脂肪含量的组合物及其用途,该组合物包含柑橘萃取物与芦荟萃取物等植物萃取物。
Description
本申请为中国专利申请号201880021260.3(发明名称:包含植物萃取物的组合物及其用途;申请日:2018年04月03日)的分案
相关申请的交叉引用
本申请主张在2017年04月03日在美国提交的美国专利申请号No.62/480,860、以及在2017年05月08日在美国提交的美国专利申请号No.62/503,185的优先权,其全部内容通过引用包含于此。
技术领域
本发明涉及一种包含植物萃取物的组合物及其用途,特别涉及一种减少脂肪生成细胞内脂肪含量的组合物及其用途。
背景技术
高油脂与高糖分的饮食方式及运动量不足使多数现代人面临肥胖问题,并且因为肥胖有较高机率罹患代谢相关疾病,例如糖尿病、高血脂症、高血压、心血管疾病、脂肪肝等,最终令身体健康受到严重威胁。还有研究显示肥胖是重要的致癌因子。此外,肥胖者较容易出现心理问题与遭遇社交障碍。因此,近年来许多医学研究皆着重于寻求抑制肥胖的方法,试图以此方式减少前述代谢疾病的发生。
抑制肥胖的方法包括饮食控制、运动、改变生活型态、药物治疗、及手术。除了严重肥胖的患者需要手术,一般大众多采饮食控制与运动的方式减重,因为现代人出于工作繁忙不易改变生活方式,又出于崇尚自然疗法而不愿意使用非必要的药物。然而,饮食控制严格要求个人的饮食均衡与热量摄取,不适当的运动方式甚至会造成身体损伤。此外,这二种方法对减重的效果有限,因为它们并非直接针对脂肪细胞,特别是内脏中的脂肪组织。
有鉴于此,开发一种便于大众使用又能有效减少脂肪细胞内脂肪含量的组合物,以预防肥胖及降低前述多种代谢疾病发生的可能性,实有其必要。
发明内容
因此,本发明的一目的在提供一种包含植物萃取物的组合物,其包含选自由柑橘萃取物与芦荟萃取物、柑橘萃取物与大蒜萃取物、柑橘萃取物与枸杞萃取物所组成的组中的任一组合。
在本发明的一实施例中,上述包含柑橘萃取物的组合物包含下列任一组合:至少1mg/ml的柑橘萃取物、及至少1mg/ml的芦荟萃取物、大蒜萃取物、或枸杞萃取物。
本发明的另一目的在提供一种医药组合物,包含前述任一种组合物、及一药学上可接受的载体。
在本发明的一实施例中,前述医药组合物的剂型可为溶液、粉末、胶囊、或锭剂。
本发明的又一目的在提供前述任一种组合物在制备减少脂肪生成细胞内脂肪含量的医药组合物中的用途。
本发明的组合物通过混合特定植物萃取物或混合特定植物萃取物与源自植物的化合物,大幅减少脂肪生成细胞内的脂肪含量,因此具备降低体脂肪及预防肥胖相关疾病发生的潜力。换言之,对个体施予有效量的本发明组合物能减少该个体的脂肪量。故,本发明的组合物可用于制备减少脂肪生成细胞内脂肪含量的医药组合物。
附图说明
图1显示本发明一实施例中不同组合物对脂肪细胞的相对脂肪含量的影响。
图2显示本发明一实施例中不同组合物对脂肪细胞的相对脂肪含量的影响。
具体实施方式
以下将配合附图进一步说明本发明的实施方式,下述所列举的实施例用以阐明本发明的发明特点及应用,而非以限定本发明的范围,对于本领域技术人员来说,在不脱离本发明的精神和范围内,还可以做出若干改进和润饰,因此本发明的保护范围应当以本申请权利要求书的内容为准。
定义
本文中所使用数值为近似值,所有实验数据皆表示在20%的范围内,优选为在10%的范围内,最优选为在5%的范围内。
本文中所谓“脂肪生成细胞”指具有合成及储存中性脂肪(neutral fat)功能的任何哺乳动物细胞,包括已分化的成熟脂肪细胞及新生的脂肪细胞。
本文中所谓「药学上可接受的载体」指一种或多种固态或液态的介质,其对哺乳动物不具毒性且不影响一组合物中活性成分的生物活性。
本发明提供一种减少脂肪生成细胞内脂肪含量的组合物,其包含多种植物萃取物或植物萃取物与源自植物的化合物。该组合物通过混合红茶、绿茶、普洱茶、四季春茶、红酒、青咖啡豆、蓝莓、柑橘、菠菜、青花菜(Brassica oleracea var.italica)苗、红花苜蓿、芦荟、迷迭香、大蒜、胡椒、姜黄、枸杞、或人参的萃取物,或混合前述植物萃取物与苹果多酚、β胡萝卜素、或番茄红素而制得。以下实施例中揭露该组合物能大幅减少脂肪生成细胞内的脂肪含量。
材料与方法
材料
DMEM培养基(Dulbecco's modified Eagle’s medium)、胎牛血清(fetal bovineserum,FBS),青霉素/链霉素(penicillin/streptomycin)、及磷酸缓冲盐溶液(phosphatebuffered saline,PBS)购自Gibco买。油红O(oil-red O)购自Sigma。甲醛(formaldehyde)及异丙醇(isopropanol)购自Echo chemical。苹果多酚(apple polyphenols)购自佶旺有限公司。β胡萝卜素(beta carotene)购自巨研贸易有限公司。番茄红素(lycopene)购自湖南朗林生物资源股份有限公司。
油红O染色及定量分析
细胞中的中性脂肪含量是以油红O染色定量。染色前,使用PBS溶液清洗细胞2次,再以10%甲醛于室温固定细胞30分钟。该固定的细胞经PBS溶液清洗2次后,以60%异丙醇处理1分钟。其后,在室温下以油红O染色剂(1.8mg/ml油红O溶于60%异丙醇水溶液)对细胞染色1小时,再以60%异丙醇处理细胞5秒以移除多余染色剂。最终,将100%异丙醇添加至染色后细胞并震荡反应10分钟以溶解细胞内染色剂,使用ELISA检测仪(enzyme-linkedimmunosorbent assay reader(酶联免疫吸附检测仪);BioTek)测量该细胞混合物在510nm的吸光度。统计上显著差异是以Excel软件的学生t检定判定。
实施例1植物萃取物的制备
实施例1-1红茶萃取物
本实施例举例说明红茶(black tea)萃取物的制备方法。首先,将红茶茶叶(fermented leaves of Camellia sinensis)洗净及干燥,以粉碎机将茶叶粗碎。其次,以水为溶剂对红茶粗碎物进行萃取,该溶剂与红茶粗碎物以液固比5~20:1~5均匀混合,萃取温度为50℃~100℃,优选为75℃~95℃。萃取时间约为0.5~3小时。经该萃取步骤所得红茶萃取物冷却至室温后,经由400目(mesh)的滤网过滤以移除残余固体物。该过滤后的红茶萃取物可进一步在45℃~70℃进行减压浓缩而获得浓缩产物。
实施例1-2绿茶萃取物
本实施例举例说明绿茶(green tea)萃取物的制备方法。首先,将绿茶茶叶(unfermented leaves of Camellia sinensis)洗净及干燥,以粉碎机将茶叶粗碎。其次,以水为溶剂对绿茶粗碎物进行萃取,该溶剂与绿茶粗碎物以液固比5~20:1~5均匀混合,萃取温度为介于50℃~100℃,优选为75℃~95℃。萃取时间约为0.5~3小时。经该萃取步骤所得绿茶萃取物冷却至室温后,经由400目的滤网过滤以移除残余固体物。该过滤后的绿茶萃取物可进一步在45℃~70℃进行减压浓缩而获得浓缩产物。
实施例1-3普洱茶萃取物
普洱茶(Puerh tea)萃取物是对普洱茶茶叶(post-fermented leaves ofCamellia sinensis)进行萃取而获得,该萃取物可购自南京泽朗生物科技有限公司。
实施例1-4四季春茶萃取物
本实施例举例说明四季春茶(Four Seasons Spring tea)萃取物的制备方法。首先,将四季春茶茶叶(leaves of Four Seasons Spring tea plant)洗净及干燥,以粉碎机将茶叶粗碎。其次,以水为溶剂对四季春茶粗碎物进行萃取,该溶剂与四季春茶粗碎物以液固比5~20:1~5均匀混合,萃取温度为50℃~100℃,优选为75℃~95℃。萃取时间约为0.5~3小时。经该萃取步骤所得四季春茶萃取物冷却至室温后,经由400目的滤网过滤以移除残余固体物。该过滤后的四季春茶萃取物可进一步在45℃~70℃进行减压浓缩而获得浓缩产物。
实施例1-5红酒萃取物
红酒萃取物是对红葡萄酒进行萃取而获得,该萃取物可购自上海博优堂生物技术有限公司。
实施例1-6青咖啡豆萃取物
青咖啡豆萃取物是对未经烘焙的咖啡属植物(Coffea spp.)种子进行萃取而获得,该萃取物可购自ARJUNA NATURAL EXTRACTS有限公司(印度)。
实施例1-7蓝莓萃取物
蓝莓萃取物是对北美蓝莓(Vaccinium Cyanococcus)果实进行萃取而获得,该萃取物可购自元弘草本科研有限公司。
实施例1-8柑橘萃取物
柑橘萃取物是对柑橘(Citrus reticulata)果实进行萃取而获得,该萃取物可购自罗顿贸易有限公司。
实施例1-9菠菜萃取物
菠菜萃取物是对菠菜(Spinacia oleracea)进行萃取而获得,该萃取物可购自弘祥农产加工厂。
实施例1-10青花菜苗萃取物
青花菜苗萃取物是对青花菜(Brassica oleracea var.italica)幼苗进行萃取而获得,该萃取物可购自蝶理株式会社。
实施例1-11红花苜蓿萃取物
红花苜蓿萃取物是对红花苜蓿(Trifolium pretense)进行萃取而获得,该萃取物可购自茂世兴业有限公司。
实施例1-12芦荟萃取物
芦荟萃取物是对芦荟(Aloe vera)进行萃取而获得,该萃取物可购自AmbePhytoextracts私人有限公司(印度)。
实施例1-13迷迭香萃取物
迷迭香萃取物是对迷迭香(Rosmarinus officinalis)进行萃取而获得,该萃取物可购自佳景百草股份有限公司。
实施例1-14大蒜萃取物
大蒜萃取物是对大蒜(Allium sativum)鳞茎进行萃取而获得,该萃取物可购自长沙市惠瑞生物科技有限公司。
实施例1-15胡椒萃取物
胡椒萃取物是对胡椒(Piper nigrum)果实进行萃取而获得,该萃取物可购自茂世兴业有限公司。
实施例1-16姜黄萃取物
姜黄萃取物是对姜黄(Curcuma longa)根茎进行萃取而获得,该萃取物可购自ARJUNA NATURAL EXTRACTS有限公司(印度)。
实施例1-17枸杞萃取物
枸杞萃取物是对枸杞(Lycium chinense)果实进行萃取而获得,该萃取物可购自湖南华康生物科技股份有限公司。
实施例1-18人参萃取物
人参萃取物是对人参(Panax ginseng)根部进行萃取而获得,该萃取物可购自湖南华诚生物资源有限公司。
实施例2
含植物萃取物的组合物降低脂肪细胞内的脂肪含量
为检验本发明的组合物对脂肪胜成细胞内脂肪储存量的影响,利用油红O染色分析由小鼠间质细胞株OP9(ATCC CRL-2749)分化而得的脂肪细胞经植物萃取物或其组合处理后,细胞内脂肪含量的变化。简言之,将OP9细胞以8×104个细胞/孔接种于含有500μl前脂肪细胞扩充培养基(90%DMEM培养基,20%FBS,1%青霉素/链霉素)的24孔培养盘,在37℃下培养7天。细胞培养期间每隔3天更换培养基为脂肪细胞分化培养基(90%DMEM培养基,20%FBS,1%青霉素/链霉素)。7天后,以显微镜(ZEISS;放大倍率400x)观察细胞内油滴形成以确认其完全分化为脂肪细胞。其后,将表1所列植物萃取物或含植物萃取物的组合物添加至细胞,于37℃下培养7-10天,期间每隔3天更换脂肪细胞分化培养基。最终,移除培养基并以PBS溶液清洗细胞,以进行油红O染色及测定各组细胞的相对脂肪含量。该相对脂肪含量是相对于对照组细胞的脂肪含量的比值(以百分比表示),而对照组的脂肪细胞是以类似方式但以不含植物萃取物的脂肪细胞分化培养基处理。
表1
表1显示脂肪细胞在不同处理后的相对脂肪含量;图1及图2是对应表1所列数值的直方图。依据表1及图1,相对于对照组,红酒萃取物、四季春茶萃取物、柑橘萃取物、菠菜萃取物、青咖啡豆萃取物、蓝莓萃取物、人参萃取物、枸杞萃取物、大蒜萃取物、芦荟萃取物、胡椒萃取物、或β胡萝卜素的单独处理无法显著减少脂肪细胞内的脂肪含量,其中有些处理甚至会显著提高脂肪含量,例如蓝莓萃取物、柑橘萃取物、菠菜萃取物(参见比较组)。此外,红茶萃取物、绿茶萃取物、普洱茶萃取物、姜黄萃取物、或苹果多酚的单独处理仅能使相对脂肪含量略下降至约90%以上。
然而,蓝莓萃取物与红茶萃取物或绿茶萃取物的组合明显降低脂肪细胞的相对脂肪含量至分别为67.50%及70.9%(参见实验组)。同样地,红酒萃取物与红茶萃取物、绿茶萃取物、普洱茶萃取物、四季春茶萃取物、柑橘萃取物、菠菜萃取物、或青咖啡豆萃取物的组合也明显降低脂肪细胞的脂肪含量。此外,柑橘萃取物与姜黄萃取物、青花菜苗萃取物、人参萃取物、枸杞萃取物、大蒜萃取物、红花苜蓿萃取物、芦荟萃取物、迷迭香萃取物、胡椒萃取物、苹果多酚、β胡萝卜素、或番茄红素的组合亦造成明显较低的脂肪含量。再者,菠菜萃取物与姜黄萃取物、人参萃取物、枸杞萃取物、大蒜萃取物、红花苜蓿萃取物、迷迭香萃取物、苹果多酚、或β胡萝卜素的组合也明显降低脂肪细胞的脂肪含量。具有上列特定组合的组合物皆意外地较单一组分简单相加表现出更高的降脂能力。
综上所述,本发明的组合物通过混合特定植物萃取物或混合特定植物萃取物与源自植物的化合物,大幅减少脂肪生成细胞内的脂肪含量,因此具备降低体脂肪及预防肥胖相关疾病发生的潜力。故,本发明的组合物可连同药学上可接受的载体用于制备减少脂肪生成细胞内脂肪含量的医药组合物。该医药组合物的剂型,可为但不限于溶液、粉末、胶囊、或锭剂。
当然,本发明还可有其它多种实施例,在不背离本发明精神及其实质的情况下,熟悉本领域的技术人员可根据本发明作出各种相应的改变和变形,但这些相应的改变和变形都应属于本发明权利要求的保护范围。
Claims (5)
1.一种包含植物萃取物的组合物,其特征在于包含选自由柑橘萃取物与芦荟萃取物、柑橘萃取物与大蒜萃取物、柑橘萃取物与枸杞萃取物所组成的组中的任一组合。
2.如权利要求1所述的组合物,其特征在于包含下列任一组合:至少1mg/ml的柑橘萃取物、及至少1mg/ml的芦荟萃取物、大蒜萃取物、或枸杞萃取物。
3.一种医药组合物,其特征在于包含权利要求1或2所述的组合物、及药学上可接受的载体。
4.如权利要求3所述的医药组合物,其特征在于具有溶液、粉末、胶囊、或锭剂的剂型。
5.一种权利要求1至4中任一项所述的组合物在制备减少脂肪含量的组合物中的用途。
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