CN115252729A - 一种治疗糖尿病的中药组合物及其应用 - Google Patents
一种治疗糖尿病的中药组合物及其应用 Download PDFInfo
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Abstract
本发明提供了一种治疗糖尿病的中药组合物,按照重量份计,配方中的药物成分包括:积雪草15‑25份、黄芪8‑12份、白花蛇舌草8‑12份、绞股蓝8‑12份、薏苡仁8‑12份、制何首乌8‑12份、桑枝8‑12份、玄参8‑12份、灵芝8‑12份、盐菟丝子12‑18份、淫羊藿12‑18份、石榴皮8‑12份、炒鸡内金8‑12份、木香8‑12份;优选的还含有无尾果15‑25份、虎耳草8‑12份、鹅掌藤8‑12份三种特殊成分。本发明的配伍不同于现有配方,本发明的产品经过了药理药效试验验证,可充分发挥复方临床用药的协同优势,有效控糖并对糖尿病患者的并发症有良好的修复作用。
Description
技术领域
本发明涉及中药制剂领域,具体涉及一种治疗糖尿病的中药组合物。
背景技术
糖尿病是一种以血浆葡萄糖水平增高为特征的内分泌代谢紊乱疾病,其与癌症、心 血管疾病并称为世界性三大疾病。2021年世界糖尿病联盟报道,全球有5.37亿成年人(20~79岁)患有糖尿病,预计到2030年,糖尿病患病人数将上升至6.43亿;2045年, 将上升至7.84亿。我国约有1.14亿糖尿病患者,是世界上糖尿病患者最多的国家。我 国成人糖尿病患病率显著上升,已达到10.4%,且发病日趋年轻化,农村人群患病率增 长快速。
糖尿病主要分为I型和II型,I型糖尿病是一种导致胰岛β细胞破坏的自身免疫性疾病,占糖尿病患者的5%~10%,主要依赖胰岛素(INS)药物治疗,长效缓释INS制 剂是各大制药公司研发重点;90%为II型糖尿病(T2DM),由基因和环境共同导致, 胰岛素抵抗(IR)是其主要病理机制。
目前临床医学中通常认为,造成II型糖尿病重要的病理生理学变化为β细胞分泌功 能受损、肝糖产生过多和胰岛素抵抗等。一般临床医学中认为,在疾病的不同阶段,不同患者的这两种因素的重要性不同。大部分与糖代谢异常密切相关的病理状态,均伴有 胰岛素抵抗,留非酒精性脂肪肝、代谢综合征、IGT和IFG和多囊卵巢综合症。目前, 临床医学中大多数研究认为,部分II型糖尿病患者发生胰岛素抵抗前,胰岛β细胞需分 泌更多的胰岛素进行代偿,以维持患者体内血糖水平的稳定。如代偿性胰岛素分泌,不 能实现与胰岛素的抵抗抗衡,患者就会出现血糖升高症状,引发糖尿病。正常人体内胰 岛素可实现对人体肝脏内产生葡萄糖有效输出的抑制,同时,可加强外周组织葡萄糖的 摄取,可维持正常水平的血浆葡萄糖指数。而糖尿病患者具有较高的胰岛素抵抗,致使 肝脏内葡萄糖的输出产生抑制缺陷,刺激外周组织,对人体葡萄糖的摄取功能造成影响。
针对T2DM治疗,西药降糖作用效果明显,但长期使用易引起酮酸中毒、肥胖、 低血糖、心血管疾病等多种不良反应。糖尿病在中医中被称为“消渴症”,最早在《黄帝 内经》及《灵枢》中均有对“消渴症”的记载。中医理论中认为消渴症的成病机理:主 要为素体阴虚且五脏柔弱,或者由于患者饮食没有节制,过多食用油腻食品或甜食导致 患者肺胃燥热,或由于患者劳欲过度导致其肾阴亏虚,病症多以阴虚为本而燥热为标。 中医中药在糖尿病的治疗和预防方面有其独特优势,并显示出了良好的发展前景,现代 药理学显示,苦味中药具有多种降糖机制,且同时又能预防糖尿病的并发症,改善糖 尿病的症状,其作用机理可能与调节细胞葡萄糖自身平衡、保护胰岛β细胞、恢复胰 岛β细胞功能、抗氧化应激、改善胰岛素抵抗、改善机体糖代谢、脂代谢、抗氧化自 由基损伤等有关。机制互补,减毒增效的降糖复方制剂运用成为糖尿病药物的治疗趋势。
2000年FDA批准施贵宝公司研发的格列本脲和盐酸二甲双胍复合片是第1上市 的西药降糖复方制剂。近年来大型药物公司加强对降糖复方制剂研发,随着西药复方降 糖药物不断临床获批上市,显示降糖复方制剂对基因与环境共同影响的糖尿病长期治疗 更为有效。中药复方含有多种生物碱类、黄酮类、萜类和多糖类降糖活性成分,通过调 节糖脂代谢、炎症通路、氧化应激水平、氨基酸代谢,修复与再生胰岛β细胞等多途 径多环节增强Ins敏感性的作用特点,降糖机制协同互补,强强联合,减毒增效。中药 复方制剂已经成为国内糖尿病药物研发的重点。
现有技术中CN105616955A一种治疗糖尿病肾病的中药。该组合物含有以下原料:野菊花、蚶壳草(积雪草)、甘草、丹参、黄芪、桑枝、桑椹子(桑椹)、三七、山药、 野灵芝、黄连、五味子、石膏、生地(地黄)、玉竹、茯苓和地骨皮;将上述药物加水 煎煮,滤去药渣,即得汤剂;口服。该组合物可用于治疗糖尿病肾病。
授权公告号CN 103110815 B(申请号201310038151.1)的专利公开了一种治疗Ⅱ型 糖尿病的中药组合物的制备方法,取下述重量配比的原料药:黄芪10-30份、西洋参 9-30份、地黄12-30份、山药15-30份、玉竹6-12份、枸杞子5-15份、天冬10-20份、 玄参10-15份、女贞子6-15份、葛根7-15份、苦瓜粉20-50份、乌药3-9份、桑椹9-15份、 黄芪15-30份;制备步骤如下:①将黄芪、西洋参、地黄、山药、玉竹、枸杞子、天冬、 玄参、女贞子、葛根、乌药、桑椹和黄芪放入砂锅中用冷水浸泡20-40分钟;②将泡好 的药材用大火煎开,转小火煎25-35分钟,滤出药液;③滤出药液后,再次加入凉水煎 开,转小火煎20-40分钟,关火前一分钟加入苦瓜粉,滤出药液,与②中药液混合,均 分成两份。虽然在治疗II型糖尿病方面取得了很好的疗效。
现有技术的产品各有其优势,然而研发一种配伍不同、疗效优良的,对糖尿病患者的并发症有良好的修复作用的治疗糖尿病的中药组合物具有重要意义。
发明内容
本发明的目的是提供一种治疗II型糖尿病的中药组合物,配伍不同于现有技术,经 过了药理药效试验验证,可充分发挥复方临床用药优势,有效控糖并对糖尿病患者的并发症有明显修复作用。
本发明的技术方案是:
一种治疗糖尿病的中药组合物,包括灵芝和黄芪,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分包括;积雪草15-25份、黄芪8-12份、白花蛇舌草8-12份、绞股蓝8-12份、薏苡仁8-12份、制何首乌8-12份、桑枝8-12份、玄参8-12份、 灵芝8-12份、盐菟丝子12-18份、淫羊藿12-18份、石榴皮8-12份、炒鸡内金8-12份、 木香8-12份。
配方2:配方中的药物成分包括;积雪草15-25份、无尾果15-25份、黄芪8-12份、 白花蛇舌草8-12份、绞股蓝8-12份、薏苡仁8-12份、制何首乌8-12份、桑枝8-12份、 玄参8-12份、灵芝8-12份、盐菟丝子12-18份、淫羊藿12-18份、石榴皮8-12份、炒 鸡内金8-12份、木香8-12份、虎耳草8-12份、鹅掌藤8-12份。
优选的,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分为;积雪草20份、黄芪10份、白花蛇舌草10份、绞股 蓝10份、薏苡仁10份、制何首乌15份、桑枝10份、玄参10份、灵芝10份、盐菟丝 子15份、淫羊藿15份、石榴皮10份、炒鸡内金10份、木香10份。
配方2:配方中的药物成分为;积雪草20份、无尾果20份、黄芪10份、白花蛇舌 草10份、绞股蓝10份、薏苡仁10份、制何首乌15份、桑枝10份、玄参10份、灵芝 10份、盐菟丝子15份、淫羊藿15份、石榴皮10份、炒鸡内金10份、木香10份、虎 耳草10份、鹅掌藤10份。
优选的,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分为;积雪草15份、黄芪12份、白花蛇舌草12份、绞股 蓝8份、薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝10份、盐菟丝 子14份、淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份。
配方2:配方中的药物成分为;积雪草15份、无尾果22份、黄芪12份、白花蛇舌 草12份、绞股蓝8份、薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝 10份、盐菟丝子14份、淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份、虎耳 草8份、鹅掌藤12份。
本发明的配方中配方1为普通配方,可以实现本发明,配方2为优选配方,配方2 中有三味成分无尾果、虎耳草、鹅掌藤在本配方中协同作用明显,效果优于配方1。
进一步的,制备方法包括以下步骤:按权利要求1或2的配方,加水煎煮2-3次, 每次加10-12倍水煎煮2-3小时,合并水煎液过200-300目滤布,离心,离心液减压浓 缩至相对密度为1.20~1.30的稠膏,真空干燥,加入糊精,粉碎成细粉,混合均匀,制 粒,干燥,整粒。
优选的,所述减压浓缩的温度为60~70℃,真空度为-0.06~-0.08Mpa。
优选的,所述真空干燥为在温度为60~70℃,真空度为-0.06~-0.08Mpa的条件下干 燥至至水分<5%。
本发明中的各原料药材的功效和药理作用如下:
积雪草:苦、辛,寒。归肝、脾、肾经。清热利湿,解毒消肿。用于湿热黄疸、 中暑腹泻、石淋血淋、痈肿疮毒、跌扑损伤。现代药理研究表明,积雪草苷和积雪草酸 可通过增加SOD的活力,减少MAD的产生,进而减轻高糖诱导的氧化应激损伤, 还可以降低BUN、UAER、MAD、SCR的水平抑制糖原的分解。
无尾果:味苦、辛,性凉。入肝、肾经。平肝息风、活血调经、清热解毒、行气止 痛。主治高血压病、头痛发热、肝炎、子宫出血、月经不调。
黄芪:甘,微温。归肺、脾经。补气升阳、固表止汗、利水消肿、生津养血、行滞 通痹、托毒排脓、敛疮生肌。其活性成分黄芪多糖可通过调控AKT/VEGF信号转导通 路,减少炎症递质的释放,黄芪甲苷可通过减少脂质过氧化,激活磷脂酰肌醇-3-激酶 AKT信号通路,调节血糖。
绞股蓝:味苦,性寒,归肺、脾、肾经。益气安神,清热降压。现代药理研究表明, 绞股蓝含有皂苷、黄酮类、糖类、萜类等多种化学成分,其主要药效成分绞股蓝皂甙具 有广泛的药理作用,如降血脂、抗肿瘤、保护肝脏、预防衰老、提高免疫力等。
白花蛇舌草:甘、淡,凉。入胃、大肠、小肠经。清热解毒,利尿消肿,活血止 痛。白花蛇舌草的活性成分主要为山柰酚、槲皮素及其糖苷,具有抗肿瘤、抗氧化、抗 菌、免疫调节和保肝等作用。对于热盛伤阴所致的消渴之上消、中消能起到清热利湿的 作用。
绞股蓝:性凉,味苦、微甘。归肺、脾、肾经。益气健脾、化痰止咳、清热解毒。 现代药理研究表明,绞股蓝的活性成分绞股蓝皂苷、多糖、黄酮具有降血糖、调脂等作 用。
薏苡仁:甘、淡,凉。归脾、胃、肺经。利水渗湿、健脾止泻、除痹、排脓、解毒 散结。其活性成分薏苡仁蛋白依赖IKK/NF-κB通道控制炎症及改善II型糖尿病胰岛素 抵抗作用。
制何首乌:苦、甘、涩,微温。归肝、心、肾经。补肝肾、益精血、乌须发、强筋 骨、化浊降脂。何首乌的水提物、乙醇提取物、正丁醇提取物、均可使血糖、FINS、 HOMA-瓜水平较模型组显著下降(P<0.01),ISI水平显著上升(P<0.01);肝细胞膜 上InsR基因表达量上升。何首乌提取物改善II型糖尿病大鼠的胰岛素抵抗,可能与上 调InsR基因表达量有关。
桑枝:微苦,平。归肝经。祛风湿,利关节。用于风湿痹病,肩臂、关节酸痛麻木。 现代药理学表明,桑枝总生物碱具有作用靶点清晰、物质基础明确、质量可控和降血糖 作用显著等特点。明确了其对α葡萄糖苷酶具有极强的抑制活性,且靶点更为精准作用 于双糖酶。
玄参:甘、苦、咸,微寒。归肺、胃、肾经。清热凉血、滋阴降火、解毒散结。玄 参多糖能够活化T2DM大鼠肝脏中IRS-2/PI3K/Akt信号通路,同时上调PPAR-γ和 GLUT-4蛋白的表达水平来改善T2DM大鼠糖脂代谢。
灵芝:甘,平。归心、肺、肝、肾经。补气安神,止咳平喘。用于心神不宁、失眠 心悸、肺虚咳喘、虚劳短气、不思饮食。现代药理研究表明,灵芝多糖GL-PS能直接 促进β细胞分泌胰岛素,来改善大鼠高血糖的症状。
盐菟丝子:辛、甘,平。归肝、肾、脾经。补益肝肾、固精缩尿、安胎、明目、 止泻;外用消风祛斑。用于肝肾不足、腰膝酸软、阳痿遗精、遗尿尿频、肾虚胎漏、胎 动不安、目昏耳鸣、脾肾虚泻。现代药理研究表明,菟丝子多糖显著降低了糖尿病大鼠 胆固醇和甘油三酯含量,表明能改善糖尿病大鼠的脂代谢紊乱。
淫羊藿:辛、甘,温。归肝、肾经。补肾阳、强筋骨、祛风湿。用于肾阳虚衰、 阳痿遗精、筋骨痿软、风湿痹痛、麻木拘挛。其有效成分淫羊藿苷能增加脑血管血流量、 调节机体免疫功能、延缓衰老。淫羊藿及其主要成分淫羊藿昔灌胃治疗的大鼠与对照组 相比,学习记忆能力明显改善,其机制可能与淫羊藿苷提高海马组织中AchE及ChAT 的表达,并且淫羊藿苷可使海马组织中超氧化物歧化酶和备脱甘肤过气氧氢化物酶活性 升高,也可有助于改善认知功能。
石榴皮:酸、涩,温。归大肠经。涩肠止泻、止血、驱虫。用于久泻、久痢、便血、 脱肛、崩漏、带下、虫积腹痛。现代研究表明,石榴皮多酚提取物、安石榴甙、鞣花酸 在合适的浓度下对а-淀粉酶、а-葡萄糖苷酶以及脂肪酸合成酶具有较好的抑制作用,即 石榴皮多酚可通过抑制与肥胖相关酶的活性达到减脂目的。
炒鸡内金:甘,平。归脾、胃、小肠、膀胱经。健胃消食、涩精止遗、通淋化石。 用于食积不消、呕吐泻痢、小儿疳积、遗尿、遗精、石淋涩痛、胆胀胁痛。现代研究表 明,鸡内金多糖能有效降低糖尿病高脂血症大鼠血糖和血脂水平,并能改善其细胞免疫 功能。
木香:辛、苦,温。归脾、胃、大肠、三焦、胆经。行气止痛,健脾消食。用于胸 胁、骁腹胀痛、泻痢后重、食积不消、不思饮食。煨木香实肠止泻,用于泄泻腹痛。现 代药理研究表明,木香烃内酯能降低血糖、减轻氧化应激损伤、抑制慢性炎性反应,对 糖尿病大鼠肾脏损伤具有保护作用。
虎耳草:微苦辛,寒,肺、脾、大肠经,祛风、清热、凉血解毒。治风疹、湿疹、 中耳炎、丹毒、咳嗽吐血、肺痈、崩漏、痔疾、其主要化学成分包括黄酮类、萜类、酚 酸类、异香豆精类等化合物。具抗菌、抗炎、抗氧化、抗肿瘤等作用。
鹅掌藤:味辛,性温,舒筋活络,消肿止痛。现代药理研究表明,其活性成分为齐 墩果酸、β-谷甾醇、豆甾醇具有显著的抗炎镇痛效果。
本发明的方解如下:
本发明以创新的中医药理论为指导,以治疗“消渴、阴虚”为出发点,以“肾精亏虚、髓海不足、阴虚燥热”为指导,以《景岳全书》所云:“悉属阴虚,无论上、中、下,急宜 治肾为主,必使阴气暂充,精血渐复,则病必自愈”为基本治法,处方合理,以临床实 践为基础。方中积雪草清热利湿,解毒消肿;无尾果平肝息风,清热解毒;两者性寒, 均可入肝肾经,用为君药。盐菟丝子补肾添精益髓,阴阳并补;制何首乌补肝肾,益精 血;淫羊藿补肾阳,强筋骨;三者均可肝、肾经,共为臣药,以助君药补肝肾之功。黄 芪益气、升阳、固卫,与玄参滋阴而共奏益气养阴之效,二者相伍,且有降血糖、尿糖 的双效作用;绞股蓝、薏苡仁,益气健脾,补肺润燥,绞股蓝配薏苡仁降血糖,而且作 用持久;桑枝、白花蛇舌草,清热利湿,活血止痛,对热盛伤阴所致的消渴之上消、中 消能起到清热利湿的功效。灵芝补气安神,以滋黄芪补气之功;炒鸡内金健胃消食,涩 精止遗,石榴皮、木香涩肠止泻,行气止痛,健脾消食。脾胃健运,则清升浊降,津液 化生有源;虎耳草、鹅掌藤清热止痛消肿;诸药共为佐使。现代药理研究证明,方中无 尾果、积雪草、黄芪、菟丝子、淫羊藿、何首乌、绞股蓝、玄参等都有较好的降低血糖、 尿糖的作用,故用之临床,确有卓效。本方配伍着眼先天兼顾脾肺,共奏滋阴补肾、益 气养阴、清热活血、降糖克消之功。
与现有技术相比,本发明的优势是:
1、本发明的中药组合物通过实验验证:对糖尿病足模型大鼠的影响对比,本发明的中药组合物各剂量组能不同程度控制糖尿病足溃疡炎症递质的过度释放,升高生长因子水平,加速创面愈合,提示本发明的中药组合物对糖尿病足模型大鼠有一定的治疗作用。本发明的中药组合物低剂量组与二甲双胍效果相当,中、高剂量组效果明显优于二 甲双胍组。提示本发明的中药组合物对糖尿病足创伤的修复效果比二甲双胍更佳。
2、通过临床实验:特殊组与二甲双胍组疗效接近,普通组和特殊组与二甲双胍对比不良反应少很多,适宜长期服用。
3、本发明配方中,无尾果、虎耳草、鹅掌藤在本配方中协同作用明显。
具体实施方式
以下结合具体实施方式对本发明的详细结构作进一步描述。
实施例1配方和制备方法
配方(按重量份计算):积雪草20份、无尾果20份、黄芪10份、白花蛇舌草10 份、绞股蓝10份、薏苡仁10份、制何首乌15份、桑枝10份、玄参10份、灵芝10份、 盐菟丝子15份、淫羊藿15份、石榴皮10份、炒鸡内金10份、木香10份、虎耳草10 份、鹅掌藤10份。
制备方法:以上17味,加水煎煮二次,第一次加12倍水煎煮2小时,第二次加10 倍水煎煮2小时,合并煎液,滤过,离心,离心液减压浓缩至相对密度为1.20~1.30(60℃) 的稠膏,真空干燥,加入适量糊精,粉碎成细粉,混合均匀,制粒,干燥,整粒,总混, 制成1000g/包,即得。
通过三批中试样品的制备,并结合大生产的实际情况,本制剂的配制工艺如表1:
表1制备工艺参数
实施例2小鼠经口灌服本发明实施例1的中药组合物单次给药毒性试验
实施例2按照国家GLP规范要求实施,观察ICR小鼠经口灌服本发明实施例1的 中药组合物药液后出现的急性毒性反应和死亡情况,为其重复给药毒性试验提供参考资 料。
试验选用检疫合格的SPF级ICR小鼠40只,雌雄各半,体重18.7~21.5g,310mm×205mm×180mm笼里饲养,每笼5只。按照国际(GB14925-2010)SPF级实验动物环境 条件要求进行饲养,动物检疫和适应环境饲养3天。
本试验观察了ICR小鼠经口灌胃本发明的产品的急性毒性试验反应。选用ICR小鼠40只,雌雄各半,按性别体重随机分为2组,分别为空白对照组(纯水)、本发明的产 品组(236.5g生药/kg),每组20只。实验前禁食不禁水12小时以上,然后按40mL/kg 分别经口灌服纯水和本发明的产品,给药当日给药2次,在每次给药后0~4小时内密 切仔细观察记录各组动物的中毒表现和特点、毒性反应出现及恢复时间以及死亡情况 等,然后每天观察2次,上下午各观察一次,连续观察14天。分别于给药前(给药当 天)及给药后第4天、第7天、第10天和第14天对动物进行称重,记录动物体重变化 及死亡情况。
试验结果:对一般活动状况的影响、动物中毒症状及死亡情况;灌胃给药结束后0~ 4小时内,空白对照组和本发明的产品组小鼠自主活动、精神状态和饮食情况均未见明显异常,未见相关毒性反应及动物死亡。给药后连续观察14天,空白对照组和本发明 的产品组小鼠自主活动、精神状态和饮食情况均未见明显异常,未见相关毒性反应及动 物死亡。
对体重的影响:动物在给药前(给药当天)、给药后第4天、第7天、第10天和第 14天称重,本发明的产品组与空白对照组动物体重未见明显差异。说明ICR小鼠经口 灌服本发明的产品,对小鼠体重增长无明显影响。
试验结束时对ICR小鼠大体解剖肉眼观察结果:各器官表面和切面均未见明显异常 情况。
结论:在本试验条件下,小鼠经口灌服本发明的产品,给药体积为40mL/kg,给药当日给药2次,累积剂量为236.5g生药/kg,相当于成人临床拟用公斤体重给药量的221 倍(成人每日剂量为1.071g生药/kg),动物未见相关毒性反应及死亡。
具体试验过程和数据如下:
1.研究目的
本研究采用ICR小鼠单次灌胃给予本发明的产品药液,连续观察14天,从小鼠的单次给药毒性试验结果了解毒性反应的程度和涉及的器官,为其重复给药毒性试验的剂量设计、毒性反应指标的选择提供依据。
2.受试物(供试品)、对照品及其它溶媒、介质信息
2.1受试物(供试品)
名称:本发明的产品 批号:210601
受试物(供试品)编号:2021013 本公司代号:QXFYKL
含量:1g浸膏含生药2.443g 规格:/
溶解性:易溶于水 浓度:/
性状:棕褐色稠膏 剂型:颗粒剂
纯度:/ 有效期至:暂定2022年05月
稳定性:暂定12个月稳定 保存条件:密封,冷藏
功能主治:清热利湿,益气养阴。用于热盛伤阴所致的消渴之上消、中消,症见口渴喜饮、消谷善饥、口干口苦;用于湿热下注所致糖尿病足及糖尿病见上述症状者。
临床拟用剂量:75g生药/日
临床拟用途径:口服,一次9.5g,一日3次
临床拟用疗程:3个月
生产单位:湖南新汇制药股份有限公司
提供单位:谷医堂(湖南)健康科技有限公司
备注:本制剂的日服生药量75g,三批的干膏得率分别为19.02%、18.60%、19.71%, 平均干膏得率为19.11%,而干膏与糊精的比例为1:1,故日服药量约为 75*0.1911*2=28.65g。本制剂拟日服3次,则每次的服用量为9.5g,根据设备尺寸及服 用习惯,本制剂的包装规格拟定为每袋装9.5g,用法用量为:开水冲服。一次1袋,一 日3次;或遵医嘱。
2.2对照品
名称:纯水
3.试验方法
3.1委托单位提供的资料:
本发明的产品处方组成:实施例1的配方。本发明的产品临床拟用剂量为75g生药/日,临床拟用途径为口服。
3.2试验设计
3.2.1剂量设计
本试验采用最大给药量法,选用ICR小鼠,40只,雌雄各半,体重18.7~21.5g, 按体重性别随机分为2组,每组20只,分别为空白对照组和本发明的产品组,按40mL/kg 分别经口灌胃给予纯水和本发明的产品药液,实验前禁食不禁水12小时以上,给药当 日给药2次,每次给药间隔约6小时,详见表2。
表2试验剂量及分组设计
注:(1)每g本发明的产品浸膏含2.443g生药;(2)临床剂量倍数人体重按70kg计算。
3.2.2动物的分组与识别
根据公司规定对动物进行编号。选取检疫合格ICR小鼠40只,雌雄各半,随机分 为2组,空白对照组和本发明的产品组,各20只动物,每笼5只动物。同时在笼具上 挂上相应的笼卡,详见表3。采用鼠尾划线法标识动物。
表3动物的分组与识别
3.3受试物(供试品)及对照品
按照公司规定对受试物(供试品)进行领用、配制、标识、使用及剩余已配制受 试物(供试品)处理。
3.3.1受试物(供试品)的配制方法:称取本发明的产品浸膏原液30.5g(1.21g浸膏/mL) 进行给药(2次/天)。
3.3.2领取后受试物(供试品)的保存条件:室温、密闭。
3.3.3使用受试物(供试品)时注意事项:混匀受试物(供试品)药液,防止污染,避免意外损耗。
3.3.4使用后剩余受试物(供试品)的处理方法:剩余的受试物(供试品)当日返还受 试物(供试品)室(节假日可在节后第一个工作日返还),按照公司规定处理。
3.3.5安全防护:在给药时间需穿戴防护服、手套、口罩。
3.3.6受试物(供试品)及对照品的给药途径、方法及选择理由,详见表4。
表4受试物(供试品)及对照品的给药途径、方法及选择理由
3.4观测时间与内容
按照公司规定对动物进行观察。给药当天,尤其在每次给药后0~4小时内密切观察 并记录各组动物的中毒表现和特点、毒性反应出现及恢复时间以及死亡情况等,然后每天观察2次,上、下午各1次,连续观察14天。
观察指标包括:动物外观、行为活动、分泌物、排泄物、饮食情况、死亡情况(死 亡时间、濒死前反应)等。如出现临床症状,需增加观察次数,以便详细观察动物中毒 反应的症状,及其起始时间、严重程度、持续时间、是否可逆。观察期结束时,按照公 司规定对剩余的试验动物进行大体解剖,肉眼观察各主要脏器的表面及切面如肝脏、肾 脏、脾脏、心脏、脑、皮肤颜色、形状和大小等,大体解剖出现异常时,均应记录并对 异常器官进行组织病理学检查。
在给药前(给药当天)及给药后第4天、第7天、第10天、第14天,使用ME1002E 型电子天平(编号170)对动物体重进行称量。
4.结果与分析
4.1对一般活动状况的影响:在两次灌胃给药结束后0~4小时内,空白对照组和本发明 的产品组ICR小鼠自主活动、精神状态和饮食情况均未见明显异常。给药后继续观察14天,空白对照组和本发明的产品组ICR小鼠自主活动、精神状态和饮食情况均未见 明显异常。
4.2动物中毒症状及死亡情况:整个试验期间未见动物死亡。
4.3体重
动物在给药前(给药当天),给药后第4天、第7天、第10天和第14天称重,本 发明的产品组与空白对照组比较动物体重未见明显差异。说明ICR小鼠经口灌服本发明 的产品,对小鼠体重增长无明显影响。详见表5。
4.4大体解剖检查结果
试验结束后采用异氟烷麻醉处死所有的试验动物,并对动物进行大体解剖检查,肉 眼观察各脏器(肝脏、肾脏、脾脏、心脏、脑、皮肤等)的位置、大小、色泽、粘连等, 并检查脏器表面和切面的质地、有无积液及肿瘤等异常改变,检查结果显示各器官表面 和切面均未见明显异常情况。
5.结论与评价
在本试验条件下,小鼠经口灌服本发明的产品,给药体积为40mL/kg,给药当日给药2次,累积剂量为236.5g生药/kg,相当于成人临床拟用公斤体重给药量的221倍(成 人每日剂量为1.071g生药/kg),动物未见相关毒性反应及死亡。
实施例3大鼠经口灌服本发明的产品(实施例1的配方产品)6个月重复给药毒性试验(方 法同实施例2)
目的:实施例3按照国家GLP规范要求实施,观察SD大鼠经口灌服给予不同剂量 本发明的产品药液多次给药毒性试验,评价本发明的产品对SD大鼠多次给药后毒性反 应,为本发明的产品临床用药提供参考资料。
试验方法:选用合格SD大鼠120只,雌雄各半,体重175.6~222.1g,475×350×200mm3笼里饲养,每笼5只。按照国际(GB14925-2010)SPF级实验动物环境条件要求进行饲养,动物检疫和适应环境饲养5天。按性别体重随机分为4组,分别为空白对照组、本 发明的产品低、中、高剂量组(11.36、22.35、44.34g生药/kg),每组30只动物(雌雄 各半),按15mL/kg体积灌胃给药,连续给药6个月(26周)。给药中期末(第13周 末)、给药末期末(第26周末)和恢复期末(第30周末)按计划分别解剖40只大鼠, 雌雄各半。检查项目包括:一般临床观察、体重、摄食量测定、血液学、血液生化、凝 血及脏器系数测定和组织病理学检查。
试验结果:试验期间,所有动物均按计划实施安乐死,试验过程无动物死亡。
一般临床观察:观察期间,本发明的产品各剂量组与同期空白对照组比较,动物外观体征、行为活动、各腔道分泌物及动物的一般状况等未出现与药物毒性相关的异常反应。
体重:与同期空白对照组比较,本发明的产品各剂量组对雄性大鼠体重均未见明显 影响,长期灌胃高剂量的药物可能会对雌性动物体重产生影响。
摄食量:与同期空白对照组比较,本发明的产品各剂量组对雄性大鼠摄食量均未见 明显影响,长期灌胃高剂量的药物可能会对雌性动物平均摄食量产生影响。
血液学检查:与同期空白对照组比较,本发明的产品各剂量组血液学指标未见有毒 理学意义的改变,结果表明本发明的产品对大鼠的血液学指标无明显影响。
血液生化检查:与同期空白对照组比较,本发明的产品各剂量组血液生化指标未见 有毒理学意义的改变,结果表明本发明的产品对大鼠的血液生化指标无明显影响。
凝血检查:与同期空白对照组比较,本发明的产品各剂量组凝血指标未见有毒理学 意义的改变,结果表明本发明的产品对大鼠的凝血指标无明显影响。
脏器系数:与同期空白对照组比较,本发明的产品各剂量组脏器系数无明显异常,结果表明本发明的产品对大鼠的脏器系数指标无明显影响。
大体解剖:给药中期、给药末期、恢复期空白对照组及本发明的产品低、中、高剂量组解剖的动物脏器表面及切面颜色、大小、质地等肉眼观察均未见明显改变。
病理检查:给药中期、给药末期、恢复期解剖的本发明的产品高剂量组动物与同期空白对照组比较,未见明显差异,未发现有毒理学意义的病理改变。
结论:在本试验条件下,SD大鼠经口灌服本发明的产品6个月,未见明显与药物 有关的毒性反应剂量(NOAEL)为44.34g生药/kg(相当于70kg成人临床拟用剂量的 41倍,等效剂量的6.6倍)。
试验数据如下表6至表14.
表6试验剂量及分组设计
注:(1)每g本发明的产品浸膏含生药2.443g;(2)临床剂量倍数人体重按70kg计算。
表7受试物(供试品)及对照品的给药途径、方法及选择理由
表8试验观测指标、内容和时间
注:与空白对照组比较,*P<0.05。
注:与空白对照组比较,*P<0.05,**P<0.01。
注:与空白对照组比较,**P<0.01。
实施例4本发明的产品药效学
目的:通过观察本发明的产品对糖尿病足模型大鼠的影响、对II型糖尿病小鼠的影响以 及对正常小鼠糖耐量的影响、为本发明的产品临床应用提供药理依据和数据支持。实验一本发明的产品对糖尿病足模型大鼠的影响
1试验目的
本试验通过复制糖尿病足大鼠模型,考察本发明的产品对糖尿病足的治疗作用,为 本发明的产品的临床应用提供药理学依据。
2试验材料
2.1受试物(供试品);本发明的产品(210601,湖南新汇制药股份有限公司)溶媒对照品:纯水,阳性对照药:二甲双胍(ABY4031,中美上海施贵宝制药有限公司):
SPF级SD大鼠雄性70只,体重150-170g,(湖南斯莱克景达实验动物有限公司,合格证号:No.430727211102038918)。SPF大小鼠维持饲料(购自北京科澳协力饲料 有限公司,生产许可证号:SCXK(京)2019-0003,包装:真空包装,每袋10kg)。
本发明的产品处方组成:实施例1的配方
临床拟用剂量:75g生药/日
临床拟用途径:口服,一次9.5g,一日3次。
临床拟用疗程:3个月
生产单位:湖南新汇制药股份有限公司
提供单位:谷医堂(湖南)健康科技有限公司
3试验方法
3.1模型制作及给药
选择检疫合格SD大鼠70只,随机分10只为空白对照组,给予正常饲料、自由饮 水;其余大鼠以高糖高脂饲喂4周,饲喂4周后,禁食12h,以链尿佐菌素(STZ)35mg/kg 腹腔注射诱导II型糖尿病模型,STZ注射5天后,取尾静脉血测量随机血糖,若大鼠空 腹血糖值大于11mmol/L时则判定为模型成功。用图章在大鼠足背部印出矩形标记 (3mm×7mm),用剪刀剪除其全层皮肤,深至筋膜,制备成糖尿病足溃疡创面模型。将 模型大鼠随机分为模型对照组、本发明的产品低、中、高剂量组、糖脉康颗粒组,每组 10只。药物组按10mL/kg灌胃相应剂量的药物,模型对照组与空白对照组灌胃等体积 纯水,连续给药2周。给药后第7,14天,以滤纸覆盖创面,得出面积数据,计算创面 愈合率,末次给药后次日,用ELISA法检测大鼠血清样本中IL-1、PDGF-BB等指标水 平,并取糖尿病足大鼠和正常大鼠溃疡创面组织进行组织病理学检查。
3.2剂量设计
本发明的产品的成人临床拟用剂量暂定为75g生药/日,按体表面积换算成大鼠临床 等效剂量=成人药量×0.018×5=6.75g生药/kg。本研究将本发明的产品大鼠等效剂量的 1倍做为低剂量,等效剂量的2倍做为中剂量,等效剂量的4倍做为高剂量。二甲双胍剂量根据文献设定为0.2g/kg,详见表15。
表15本发明的产品剂量设计表
注:每g本发明的产品浸膏含生药2.443g。
3.3药物配制方法:
二甲双胍组:取盐酸二甲双胍3粒(0.5g),碾磨后加纯水配制至150mL(0.01g/mL)低剂量组:称取本发明的产品浸膏11.05g,加纯水配制至40mL(0.276g浸膏/mL) 中剂量组:称取本发明的产品浸膏22.10g,加纯水配制至40mL(0.553g浸膏/mL) 高剂量组:称取本发明的产品浸膏44.20g,加纯水配制至40mL(1.105g浸膏/mL) 配制体积根据每周大鼠体重变化进行调整。
3.4检测指标
一般观察:造模后及给药后密切观察并记录各组动物外观体征、行为活动、分泌物、 排泄物、饮食情况等。如发现动物死亡或濒死,应及时对动物进行解剖观察。每周对动物体重进行称量。
检测指标:大鼠创面愈合情况;给药后第7,14天,以滤纸覆盖创面,得出面积数据,计算创面愈合率。创面愈合率=(原始创面面积-残余创面面积)/原始创面面积 ×100%。
相关因子水平检测:末次给药后次日,用ELISA法检测大鼠血清样本中及IL-1、PDGF-BB等指标水平。
组织病理观察:取糖尿病足大鼠和正常大鼠创面组织,常规HE染色,观察大鼠溃疡创面病理情况。
4试验结果
4.1对大鼠创面愈合率的影响
与空白对照组比较,模型对照组大鼠第7天、第14天愈合率显著降低(P<0.01); 与模型对照组比较,本发明的产品低、中、高剂量组、二甲双胍组第7天愈合率显著升 高(P<0.05或P<0.01),本发明的产品低、中、高剂量组、二甲双胍组第14天愈合率 明显升高(P<0.05或P<0.01),本发明的产品中、高剂量组第7天和第14天愈合率显 著高于二甲双胍组(P<0.05)。结果见表16。试验结果提示本发明的产品有促进糖尿病 足大鼠溃疡创面愈合的作用,与二甲双胍组相比较,本发明的产品低剂量组愈合率与二 甲双胍组相当,本发明的产品中、高剂量组愈合率明显高于二甲双胍组。
表16本发明的产品对大鼠创面愈合率的影响
注:与空白对照组比较,*P<0.05,**P<0.01;与模型对照组比较,△P<0.05,△△P< 0.01,与二甲双胍组比较,#P<0.05.。
4.2大鼠血清中IL-1、PDGF-BB水平的影响
与空白对照组比较,模型对照组动物血清IL-1水平显著升高(P<0.01),PDGF-BB水平显著降低(P<0.01);与模型对照组比较,本发明的产品中、高剂量组、二甲双胍 组动物血清IL-1水平明显降低(P<0.05或P<0.01),PDGF-BB水平明显升高(P<0.05 或P<0.01),结果见表17。结果提示本发明的产品可控制糖尿病足溃疡炎症递质的过 度释放,升高生长因子水平,加速创面愈合。本发明的产品中、高剂量组与二甲双胍组 相比较动物血清IL-1水平显著降低(P<0.05或P<0.01),PDGF-BB水平明显升高(P<0.05 或P<0.01),详见表17。
表17本发明的产品对大鼠血清中IL-1、PDGF-BB水平的影响
注:与空白对照组比较,*P<0.05,**P<0.01;与模型对照组比较,△P<0.05,△△P< 0.01,与二甲双胍组比较,#P<0.05。
5.结论
试验结果显示:在本试验条件下,本发明的产品各剂量组能不同程度控制糖尿病足 溃疡炎症递质的过度释放,升高生长因子水平,加速创面愈合,提示本发明的产品对糖尿病足模型大鼠有一定的治疗作用。本发明的产品低剂量组与二甲双胍效果相当,中、 高剂量组效果明显优于二甲双胍组。揭示本发明的产品对糖尿病足创伤的修复效果比二 甲双胍更佳。
实施例5临床数据
1.资料与方法
1.1一般资料:选取2018年1月至2021年12月到中南大学湘雅二医院以及湖南谷医堂 连锁中医门诊有限公司长沙雨花区京豪大厦门诊部就诊的患者144例,随机分为二甲双胍组、普通治疗组和特殊治疗组,每组各48例。统计基础资料:二甲双胍组年龄42~ 69(52.1±7.3)岁;病程1~10(4.2±2.0)年;性别比例男/女为28/20。普通治疗组年龄 42~70(50.6±6.7)岁;病程1~10(3.8±2.0)年;性别比例男/女为30/18。特殊治疗组 年龄42~78(51.3±7.5)岁;病程1~10年(4.4±2.0)年;性别比例男/女为29/19。各 组基础资料经由临床统计学检验,没有统计学意义(P>0.05),有较高可比性。
1.2病例纳入及排除标准
(1)纳入标准:均符合WHO有关II型糖尿病的相关诊断标准;确诊6个月以上,意识清楚、有一定认知水平;2hPG≥11.0mmol/L;FBG≥7.0mmol/L;均采用降糖药口服治疗; 均知晓本次实验且自愿参与。
1.3排除标准:哺乳期、妊娠期妇女、各种药物过敏史者、言语交流障碍者、心理疾病 者、糖尿病并发症者、慢性消化系统疾病者、合并心、肝、肾、等严重功能障碍、肢体 运动功能障碍者、合并造血系统疾病者、精神疾病者。
1.4方法:二甲双胍组患者口服二甲双胍(国药准字H20050699,规格:0.5g)(由中美上 海施贵宝制药有限公司生产),0.5g,3次/d;本发明的产品普通组和特殊组口服本发明的产品(由湖南新汇制药股份有限公司生产,由谷医堂(湖南)健康科技有限公司提 供),9.5g/次,3次/d。3组患者均连续治疗3个月。
本发明的产品普通组:积雪草15份、黄芪12份、白花蛇舌草12份、绞股蓝8份、 薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝10份、盐菟丝子14份、 淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份;
本发明的产品特殊组:积雪草15份、无尾果22份、黄芪12份、白花蛇舌草12份、 绞股蓝8份、薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝10份、盐 菟丝子14份、淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份、虎耳草8份、 鹅掌藤12份。
2.制备方法:同实施例1。
3.观察指标及评价标准
3.1观察指标:比较三个组别HbAlc(糖化血红蛋白)、2hPG(餐后2h血糖)、FBG (空腹血糖)等水平变化、总有效率及不良反应率。
3.2总有效率评价标准
3.21显效:HbAlc、2hPG、FBG等分别为≤7.0%、≤8.2mmol/L、≤7.0mmol/L。
3.22有效:HbAlc、2hPG、FBG等分别为≤5.0%、≤10.0mmol/L、≤8.3mmol/L。
3.23无效:未达到上述指标。
统计学分析:采用统计学软件进行数据分析,计数资料以[例(%)]表示,行X2检验; 计量资料以(均数±标准差)表示,进行t检验。以P<0.05为差异有统计学意义。
4.结果
1、血糖水平观察对比:普通治疗组与特殊治疗组治疗后HbAlc、2hPG、FBG等水 平接近二甲双胍组,差异无统计学意义(P>0.05),见表18。
2、临床有效率对比
(1)二甲双胍组与普通治疗组有效率对比:普通治疗组总有效率少于二甲双胍组。差 异有统计学意义(P<0.05),见表19。
表19二甲双胍组与普通治疗组总有效率观察对比[例(%)]
(2)二甲双胍组与特殊治疗组总有效率对比:特殊治疗组总有效率与二甲双胍组相 当。差异无统计学意义(P>0.05),见表20。
表20二甲双胍组与特殊治疗组总有效率观察对比[例(%)]
(3)普通治疗组与特殊治疗组总有效率对比:普通治疗组与特殊治疗组有效率对比, 特殊治疗组有效率优于普通组,差异有统计学意义(P<0.05),见表21。
表21普通治疗组与特殊治疗组总有效率观察对比[例(%)]
3、不良反应观察对比:普通治疗组与特殊治疗组的胃肠道反应发生率、浮肿发生率、低血糖发生率均低于二甲双胍组。差异有统计学意义(P<0.05),见表22、表23。
表22二甲双胍组与普通治疗组不良反应率观察对比[例(%)]
表23二级双胍组与特殊治疗组不良反应率观察对比[例(%)]
5.结论
本发明的产品治疗糖尿病,能有效降低糖尿病患者血糖水平,有显著治疗效果且大 幅降低药物不良反应,具有较高安全性。
以上所述为本发明的具体实施方式,但本发明的保护范围不局限于此,任何熟悉技 术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其构思加以等 同替换或改变,都应涵盖在本发明权利要求的保护范围之内。
Claims (7)
1.一种治疗糖尿病的中药组合物,包括灵芝和黄芪,其特征是,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分包括;积雪草15-25份、黄芪8-12份、白花蛇舌草8-12份、绞股蓝8-12份、薏苡仁8-12份、制何首乌8-12份、桑枝8-12份、玄参8-12份、灵芝8-12份、盐菟丝子12-18份、淫羊藿12-18份、石榴皮8-12份、炒鸡内金8-12份、木香8-12份;配方2:配方中的药物成分包括;积雪草15-25份、无尾果15-25份、黄芪8-12份、白花蛇舌草8-12份、绞股蓝8-12份、薏苡仁8-12份、制何首乌8-12份、桑枝8-12份、玄参8-12份、灵芝8-12份、盐菟丝子12-18份、淫羊藿12-18份、石榴皮8-12份、炒鸡内金8-12份、木香8-12份、虎耳草8-12份、鹅掌藤8-12份。
2.根据权利要求1所述治疗糖尿病的中药组合物,其特征是,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分包括;积雪草20份、黄芪10份、白花蛇舌草10份、绞股蓝10份、薏苡仁10份、制何首乌15份、桑枝10份、玄参10份、灵芝10份、盐菟丝子15份、淫羊藿15份、石榴皮10份、炒鸡内金10份、木香10份;
配方2:配方中的药物成分包括;积雪草20份、无尾果20份、黄芪10份、白花蛇舌草10份、绞股蓝10份、薏苡仁10份、制何首乌15份、桑枝10份、玄参10份、灵芝10份、盐菟丝子15份、淫羊藿15份、石榴皮10份、炒鸡内金10份、木香10份、虎耳草10份、鹅掌藤10份。
3.根据权利要求1所述治疗糖尿病的中药组合物,其特征是,按照重量份计,选自以下两种配方之一:
配方1:配方中的药物成分包括;积雪草15份、黄芪12份、白花蛇舌草12份、绞股蓝8份、薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝10份、盐菟丝子14份、淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份;
配方2:配方中的药物成分包括;积雪草15份、无尾果22份、黄芪12份、白花蛇舌草12份、绞股蓝8份、薏苡仁12份、制何首乌10份、桑枝10份、玄参10份、灵芝10份、盐菟丝子14份、淫羊藿18份、石榴皮12份、炒鸡内金12份、木香8份、虎耳草8份、鹅掌藤12份。
4.根据权利要求1-3中任意一项所述治疗糖尿病的中药组合物,其特征是,制备方法包括以下步骤:按权利要求1或2的配方,加水煎煮2-3次,每次加10-12倍水煎煮2-3小时,合并水煎液过200-300目滤布,离心,离心液减压浓缩至相对密度为1.20~1.30的稠膏,真空干燥,加入糊精,粉碎成细粉,混合均匀,制粒,干燥,整粒。
5.根据权利要求4所述治疗糖尿病的中药组合物,其特征是,所述减压浓缩的温度为60~70℃,真空度为-0.06~-0.08Mpa。
6.根据权利要求4所述治疗糖尿病的中药组合物,其特征是,所述真空干燥为在温度为60~70℃,真空度为-0.06~-0.08Mpa的条件下干燥至至水分<5%。
7.权利要求1-6任意一项所述治疗糖尿病的中药组合物的应用,其特征是,所述中药组合物在制备治疗糖尿病的保健品或者药物中的应用。
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