CN109432162A - 地八角提取物及其药物组合物和制备方法与应用 - Google Patents
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Abstract
本发明涉及地八角提取物在制备抗氧化、抗肿瘤和镇痛产品中的应用,属于中药、天然药物制药领域。以本发明提取物为活性成份的抗氧化、抗肿瘤和镇痛的药物。本发明拓展了地八角药材的适应症,提高了传统地八角药材的药用价值。
Description
技术领域:
本发明属于中药、天然药物制药领域,具体地,涉及地八角提取物的制备方法和以该提取物为活性成份的药物组合,及其在抗氧化、抗肿瘤和镇痛剂中的应用。
技术背景:
地八角(Astragalus bhotanensis Baker)又称为不丹黄芪,为豆科黄芪属多年生草本植物,产云南、西藏、贵州、四川、陕西、甘肃等地;生于海拔600–2800米间的山坡、山沟、河漫滩、田边、阴湿处及灌丛下。《中华本草》等中医药典籍记载该植物全草药用,有清热解毒、利尿止泻的功效;主治咽喉肿痛、咳嗽、麻疹、浮肿、泄泻、痢疾牙痛、口鼻出血等。近几年,对地八角的研究报道主要集中在地八角作为复方组分应用于炎症和鼻咽癌的治疗,以及饲料添加剂等方面(中国专利CN 105168909A、CN 105902995A、CN 105535664A、CN 104784479A、CN106109940A、CN 101204444A、CN 105560553A、CN 104547583A、CN 105362680A、CN104547587A、CN 103652370B)。目前,国内外未见地八角提取物的制备方法和以该提取物为活性成份的药物组合,及其在抗氧化、抗肿瘤和镇痛中应用的报道。
发明内容:
本发明旨在提供地八角提取物的制备方法和以其为活性成份的药物组合物,以及它们在制备抗氧化、抗肿瘤和镇痛剂中的应用。
本发明的上述目的是通过下面的技术方案得以实现的:
地八角提取物的制备方法,取地八角植物的根或全株阴干,粉碎到30目,用有机溶剂氯仿或丙酮或甲醇或乙醇或水在室温条件下浸提2-5次,每次12-48 h,合并提取液,提取液减压浓缩得浸膏。或将地八角植物的根或全株阴干,粉碎到30目,用有机溶剂氯仿或丙酮或甲醇或乙醇或水在80–120 oC条件下回流提取2-5次,每次2-8 h,合并提取液,提取液减压浓缩得浸膏。浸泡或热回流时,可辅以超声或微波提取,每次用溶剂量为浸泡药渣重量的5–25倍。
抗氧化剂,含有地八角提取物和常规辅剂。
抗肿瘤剂,含有地八角提取物和常规辅剂。
镇痛剂,含有地八角提取物和常规辅剂。
药物组合物,其中含有治疗有效量的地八角提取物和药学上可接受的载体。
地八角提取物在制备抗氧化剂中的应用。
地八角提取物在制备抗肿瘤剂中的应用。
地八角提取物在制备镇痛剂中的应用。
所述的地八角提取物在制备治疗胃癌、肝癌、白血病的药物中的应用。
本发明用于抗氧化、抗肿瘤、镇痛的药物组合物,其中含有地八角提取物和药学上可接受的载体。
本发明药物组合物中所述药学上可接受的载体是指药学领域常规的药物载体。本发明地八角提取物可以组合物的形式通过口服、鼻吸入、直肠或肠胃外给药的方式施用于需要这种治疗的患者。用于口服时,可将其制成常规的固体制剂如片剂、粉剂、粒剂、胶囊等,制成液体制剂如油悬浮剂、糖浆、酏剂等;用于肠外给药时,可将其制成注射用的溶液等。优选的形式是片剂、胶囊和注射剂。
本发明药物组合物的各种剂型可以按照药学领域的常规生产方法制备。例如使活性成份与一种或多种载体混合,然后将其制成所需的剂型。
本发明的药物组合物优选含有重量比为0.1%–99.5%的活性成份,最优选含有重量比为0.5%–95%的活性成份。
本发明地八角提取物的施用量可根据用药途径、患者的年龄、体重、所治疗的疾病的类型和严重程度等变化,其日剂量可以是0.1–100 mg/kg体重,优选1–50 mg/kg体重。可以一次或多次施用。
本发明的地八角提取物显示出较好的抗氧化、抗肿瘤活性和镇痛活性。
本发明对地八角提取物进行了抗氧化活性筛选,地八角提取物显示较好的抗氧化活性。在抗氧化活性应用中,地八角提取物是以如下的量施用于基材或一种群上,所述量的范围0.02-15 g,优选在0.05-5 g,任选地与载体和/或媒体相结合。
本发明对地八角提取物进行了细胞毒活性筛选,地八角提取物显示较好的抗肿瘤活性。在抗肿瘤活性应用中,地八角提取物是以如下的量施用于基材或一种群上,所述量的范围0.03-20 g,优选在0.05-10 g,任选地与载体和/或媒体相结合。
本发明对地八角提取物进行了镇痛活性筛选,地八角提取物显示较好的镇痛活性。在镇痛活性应用中,地八角提取物是以如下的量施用于基材或一种群上,所述量的范围0.03-30 g,优选在0.05-10 g,任选地与载体和/或媒体相结合。
具体实施方式:
下面用本发明的实施例来进一步说明本发明的实质性内容,可以使本专业人员更全面地理解本发明,但不以任何方式限制本发明。
实施例1:
本发明地八角提取物的制备:
将地八角植物的根阴干(0.5 kg),粉碎到30目,用95%甲醇在室温条件下浸提3次,每次2.5 L、24 h,提取液合并,减压浓缩提取液得浸膏(64 g)。
实施例2:
本发明提取物的抗氧化活性检测:
采用DPPH法测定本发明提取物的抗氧化活性。在96孔板中加入50 μL不同浓度的供试品溶液,随后向各孔中快速地加入150 μL浓度为100 μM的DPPH溶液,室温黑暗处放置30min后,在酶标仪517 nm波长处测定吸光度值;实验中同时设置空白对照(50 μL DMSO +150 μL DPPH)、溶剂对照(50 μL DMSO + 150μL甲醇)及阳性对照(50 μL Vc + 150 μLDPPH)。根据以下公式计算供试品对DPPH吸光度的百分抑制率,以IC50评价其抗氧化活性。
I % = [(A blank – A sample) /A blank] × 100%
A blank为未加样的DPPH的吸光度,A sample为样品与DPPH反应后的吸光度。
活性数据见表1。
表1 地八角提取物的抗氧化活性数据(χ± s, n = 4)
实施例3:
本发明提取物的细胞毒活性检测:
采用MTT法测定本发明提取物对人胃癌细胞株(SGC-7901)、肝癌细胞株(HepG2)和人红细胞白血病细胞株(K-562)的细胞毒性。实验设阴性对照组(水)、DMSO溶剂对照组、阳性对照组(顺铂)和不同浓度的供试样品,每个浓度设5个平行。收集对数生长期细胞,血球计数板计数,按每孔4500个癌细胞量接种于96孔平底细胞培养板中,置于5%CO2、湿度90%以上、37 ℃温箱中培养。24 h后取出加入一定量的待测样品,继续培养3 d后取出置于显微镜下观察每孔细胞形态,记录细胞形态变化情况,接着每孔加入5 mg/mL的MTT溶液(溶于平衡盐溶液PBS)50 μL,37 ℃反应4 h后,将细胞培养液吸出,每孔加入100 μL DMSO将Formazane充分溶解,将细胞培养板置于MK3酶标仪上,用570 nm波长测各孔的吸光度(A),按下列公式求生长抑制率。而后以样品浓度为横坐标,以抑制率为纵坐标,作图并求出抑制率为50%时样品的浓度(IC50),样品活性结果即以半数抑制浓度(IC50)表示。活性数据见表2。
表2 地八角提取物的细胞毒活性数据
实施例4:
本发明提取物的镇痛活性检测:
采用醋酸扭体实验测定本发明提取物的镇痛活性。昆明种小鼠雌雄各半,体重18–22g,实验前12小时禁食、不禁水,随机分为对照组、样品组、阳性药组,每组6只。以200 mg/kg的剂量灌胃给药,阴性对照组给予等量生理盐水,阳性药组给予阿司匹林,给药40 min后,每只小鼠腹腔注射0.6%冰醋酸溶液0.2 mL/10g,注射冰醋酸3–5 min后观察并记录15 min内小鼠的扭体次数,按下列公式计算受试物对小鼠醋酸扭体的抑制率,以此来评价受试物的镇痛作用。
疼痛抑制率(%)=(阴性对照组扭体次数-给药组扭体次数)/阴性对照组扭体次数×100%
活性数据见表3。
表3 地八角提取物的镇痛活性数据
Claims (12)
1.地八角提取物的制备方法,其特征在于,用有机溶剂氯仿或丙酮或甲醇或乙醇或水直接冷浸或者热回流提取地八角植物的根或全株,得到提取液,将提取液浓缩、干燥得到地八角提取物。
2.权利要求1所述提取物的制备方法,其特征在于将地八角植物的根或全株阴干,粉碎到30目,用有机溶剂氯仿或丙酮或甲醇或乙醇或水在室温条件下提取,提取液浓缩得浸膏。
3.权利要求1所述提取物的制备方法,其特征在于将地八角植物的根或全株阴干,粉碎到30目,用有机溶剂氯仿或丙酮或甲醇或乙醇或水热回流提取,提取液浓缩得浸膏。
4.如权利要求2和3所述的制备方法,其特征在于,浸泡或热回流时,可辅以超声或微波提取,每次用溶剂量为浸泡药渣重量的5–25倍。
5.药物组合物,其中含有治疗有效量的权利要求1所述提取物和药学上可接受的载体。
6.抗氧化剂,其中含有权利要求1所述提取物和常规辅剂。
7.抗肿瘤剂,其中含有权利要求1所述提取物和常规辅剂。
8.镇痛剂,其中含有权利要求1所述提取物和常规辅剂。
9.权利要求1所述提取物在制备抗氧化剂中的应用。
10.权利要求1所述提取物在制备抗肿瘤剂中的应用。
11.权利要求1所述提取物在制备镇痛剂中的应用。
12.权利要求1所述提取物在制备治疗胃癌、肝癌、白血病的药物中的应用。
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