CN112494487A - 毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用 - Google Patents
毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用 Download PDFInfo
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Abstract
本发明公开了毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用，涉及医药技术领域。该毛钩藤碱对胰岛素抵抗、糖尿病及其并发症，具有一定的治疗作用，低浓度就能发挥药效，并且无细胞毒性，能够有效改善胰岛素抵抗、治疗II型糖尿病并且可以逆转肝脏脂肪变性，为临床治疗II型糖尿病提供了新的治疗方法和治疗药物。
Description
技术领域
本发明涉及医药技术领域，尤其是涉及毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用。
背景技术
糖尿病是一种以高血糖为主要特征的代谢性疾病，其中II型糖尿病占比高达90％以上，以胰岛素抵抗为中心因素，多伴有不同程度的脂肪肝、血脂异常以及高血压，动脉粥样硬化等心血管疾病并发症，严重威胁人类身心健康，给国家和个人造成沉重的经济负担，对公共卫生威胁极大。II型糖尿病多发于年长人士，并且有极大的低龄化趋势。
作为中医中药应用史中很常见的中药，钩藤已被临床经验以及现代研究证实具有抗高血压的作用，其提取物对心血管及中枢神经系统疾病具有显著疗效。其中的单体提取物毛钩藤碱也被证实具有降血压、抗心律失常，抗心肌缺血等作用与活性。
目前临床上常规使用的抗糖尿病药物主要有胰岛素、二甲双胍、磺酰脲类药物及近来上市的噻唑烷二酮类药物等，疗效虽良好，然而实践证明这些药物依然有着长期使用易产生肝毒性、体重增加、不能完全控制糖化血红蛋白水平和低血糖等问题。因此，从中药中提取更有效的天然化合物挖掘新的治疗药物成为了现在关注的热点，开发更加安全有效的抗糖尿病药物应用于临床治疗具有重要的意义。目前，尚未有关于毛钩藤碱应用于II型糖尿病的报道。
发明内容
本发明旨在至少解决现有技术中存在的问题之一。为此，本发明提供一种毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用，所述药物对糖尿病及相关疾病有良好治疗作用。
本发明还提出上述毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病和及其并发症药物相关药理途径中激活剂的应用。
根据本发明的第一方面实施方式的毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用。
根据本发明的一些实施方式，所述毛钩藤碱(Hirsutine)的结构式如下所示：
根据本发明的一些实施方式，所述毛钩藤碱分子量为368.47。
根据本发明的一些实施方式，所述毛钩藤碱由中药钩藤提取得到。
根据本发明的一些实施方式，所述糖尿病为II型糖尿病。
根据本发明的一些实施方式，所述并发症包括脂代谢紊乱疾病和/或肾病；优选地，所述脂代谢紊乱疾病为非酒精性脂肪肝。
根据本发明的一些实施方式，所述治疗胰岛素抵抗、糖尿病及其并发症药物为抑制胰岛素抵抗状态下的细胞的糖异生相关基因G6Pase、PEPCK或PGC-1α表达的药物。
根据本发明的一些实施方式，所述药物还包括药学上可接受的载体或辅料。
根据本发明的一些实施方式，所述药物的剂型为本领域常规的各种剂型，优选地为固体、半固体或液体的形式，可以为水溶液、非水溶液或混悬液，更优选地为片剂、胶囊剂、软胶囊剂、颗粒剂、丸剂、口服液、干混悬剂、滴丸剂、干浸膏剂、注射剂或输注剂。
根据本发明的一些实施方式，所述药物的给药方式可以为本领域常规的给药方式，包括但不限于注射给药或口服给药。所述注射给药可以为静脉注射、肌肉注射、腹腔注射、皮内注射或皮下注射等途径。
根据本发明第二方面实施方式的毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病和及其并发症药物相关药理途径中激活剂的应用。
根据本发明的一些实施方式，所述药理途径为PI3K/Akt/GSK3β通路。
根据本发明的一些实施方式，毛钩藤碱在制备PI3K/Akt/GSK3β信号通路激活剂中的应用。
根据本发明的一些实施方式，所述应用为激活Akt、PDK1和GSK3β蛋白表达。
根据本发明实施例的毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用，至少具有如下有益效果：本发明方案的毛钩藤碱能够用于制备改善胰岛素抵抗、糖尿病和脂肪肝药物或PI3K/Akt/GSK3β信号通路激活剂，低浓度就能发挥药效，并且无细胞毒性，能够有效改善胰岛素抵抗、治疗II型糖尿病并且可以逆转糖尿病合并的肝脏脂肪变性。
附图说明
图1为本发明实施例2中毛钩藤碱在不同时间高糖高胰岛素(中文简称高糖高胰)状态下诱导的胰岛素抵抗HepG2模型中葡萄糖的消耗量结果图，其中，&，&&分别是与con组比较p<0.05和p<0.01，*和**分别是与HGHI模型组比较p<0.05和p<0.01；
图2为本发明实施例2中在不同浓度毛钩藤碱提高胰岛素抵抗条件下HepG2细胞的葡萄糖摄取量结果图，其中，&是与con组比较p<0.05，*和**分别是与HGHI模型组比较p<0.05和p<0.01；
图3为本发明实施例2中的在不同浓度毛钩藤碱对降低胰岛素抵抗条件下HepG2细胞的肝糖输出的影响结果图，其中，&&是与con组比较p<0.01，*和**分别是与HGHI模型组比较p<0.05和p<0.01；
图4为本发明实施例2中的不同浓度毛钩藤碱增加胰岛素抵抗条件下HepG2细胞中糖原含量结果图，其中，&&是与con组比较p<0.01，*和**分别是与HGHI模型组比较p<0.05和p<0.01；
图5为本发明实施例2中的毛钩藤碱抑制与糖异生相关基因表达结果图，其中，图A为G6Pase基因表达图，图B为PEPCK基因表达图，&&代表是与con组比较p<0.01，*、**和***分别代表与HGHI模型组比较p<0.05，p<0.01和p<0.001；
图6为本发明实施例2中的毛钩藤碱抑制与糖异生相关基因表达结果图，其中，图A为PGC-1α基因表达图，图B为FoxO1基因表达图，&&代表是与con组比较p<0.01，*、**和***分别代表与HGHI模型组比较p<0.05，p<0.01和p<0.001；
图7为本发明实施例2中的毛钩藤碱对胰岛素抵抗状态下HepG2细胞中葡萄糖转运蛋白GLUT2的表达量的影响结果图；
图8为本发明实施例2中的毛钩藤碱对胰岛素抵抗状态下HepG2细胞的糖酵解能力的影响结果图，其中，图A为不同时间下细胞外酸化率图，图B为不同糖酵解程度下细胞外酸化率图，*代表与HGHI模型组比较p<0.05；
图9为本发明实施例3中的毛钩藤碱对胰岛素抵抗状态下HepG2细胞Akt，PDK1，GSK3β的磷酸化水平的影响结果图；
图10为本发明实施例3中的PI3K抑制剂抑制毛钩藤碱提高的胰岛素抵抗条件下对HepG2细胞的葡萄糖摄取量的影响结果图，其中，&代表与con组比较p<0.05，**和***分别代表与HGHI模型组比较p<0.01和p<0.001，##和###是与相应浓度的毛钩藤碱给药组比较p<0.01和p<0.001；
图11为本发明实施例3中的PI3K抑制剂抑制毛钩藤碱刺激胰岛素抵抗条件下HepG2细胞PI3K、Akt、GSK3β蛋白表达及磷酸化水平结果图；
图12为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠体重的影响结果图；
图13为本发明实施例4中的不同浓度毛钩藤碱对db/db糖尿病鼠的空腹血糖量的影响结果图，其中，**和***分别是与HGHI模型组比较p<0.01和p<0.001；
图14为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠的口服葡萄糖(1mg/kg葡萄糖灌胃浓度)耐受量的影响结果图，其中，图A为葡萄糖灌胃后不同时间下糖尿病鼠血糖值的变化图，图B为糖尿病鼠口服葡萄糖耐受量的变化结果图。###是与WT组比较p<0.001，**和***分别是与db/db组比较p<0.01和0.001；
图15为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠的胰岛素(1Ukg-1胰岛素腹腔注射浓度)耐受量的影响结果图，其中，图A为注射胰岛素后不同时间下血糖值的变化图，图B为db/db糖尿病鼠胰岛素耐受量的影响结果图，###是与WT组比较p<0.001，**和***分别是与db/db组比较p<0.01和0.001；
图16为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠肝脏的外观的影响结果图；
图17为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠的肝系数的影响结果图，###是与WT组比较p<0.001，*、**和***分别是与db/db组比较p<0.05，p<0.01和0.001；
图18为本发明实施例4中的毛钩藤碱对糖尿病鼠肝脏脂质沉积的影响结果图(上图比例尺：300μM，下图比例尺：100μM)；
图19为本发明实施例4的毛钩藤碱对糖尿病鼠肝脏脂肪肝病变程度，细胞肿胀程度以及炎症程度的影响结果图(上图比例尺：300μM，下图比例尺：100μM)；
图20为本发明实施例4中的毛钩藤碱对糖尿病鼠非酒精性脂肪肝活动度评分的影响结果图，其中，##是与con组比较p<0.01，*和**分别是与model组比较p<0.05和0.01；
图21为本发明实施例4中的毛钩藤碱对db/db糖尿病鼠胰岛细胞的影响结果图(上图比例尺：300μM，下图比例尺：50μM)；
图22为本发明实施例4中的毛钩藤碱对糖尿病鼠的肾脏的影响结果图(上图比例尺：300μM，下图比例尺：50μM)。
具体实施方式
为详细说明本发明的技术内容、所实现目的及效果，以下结合实施方式予以说明。实施例中所使用的试验方法如无特殊说明，均为常规方法；所使用的材料、试剂等，如无特殊说明，均可从商业途径得到的试剂和材料。以下实施例1-3中使用的毛钩藤碱，购买自成都瑞芬思生物科技有限公司，CAS号为7729-23-9。实施例4使用的毛钩藤碱，购买自上海诗丹德标准技术服务有限公司。
实施例1：高糖高胰岛素诱导HepG2细胞产生胰岛素抵抗
HepG2细胞是一种源于人的肝胚胎瘤细胞，保留了肝细胞的许多特性，表型与肝细胞极为相似，是研究胰岛素抵抗发病机制的理想细胞系。
以高糖高胰岛素刺激来诱导HepG2细胞36h，使该细胞表面胰岛素受体数目下降，形成胰岛素抵抗，即相比正常状态下的细胞，其葡萄糖消耗和摄取是明显受抑制的。且该胰岛素抵抗模型可稳定持续48h。
实施例2：毛钩藤碱对胰岛素抵抗型HepG2细胞内糖稳态的影响
1、实验步骤：将HepG2细胞分为6组，具体如下：(1)对照组，(2)高糖高胰岛素抵抗模型组，(3)高糖高胰岛素抵抗模型+0.01μM毛钩藤碱，(4)高糖高胰岛素抵抗模型+0.1μM毛钩藤碱，(5)高糖高胰岛素抵抗模型+1μM毛钩藤碱，(6)高糖高胰岛素抵抗模型+1mM二甲双胍。在给药后的3h，6h，12h，24h间分别收集培养基上清液，用购自南京建成的葡萄糖氧化酶试剂盒(MO10)检测培养基上清液葡萄糖含量来间接计算细胞消耗的葡萄糖量。以一种葡萄糖荧光类似物2-NBDG的荧光值来表示葡萄糖摄取量。收集HepG2细胞，使用超声波破碎细胞，再通过使用购自北京索莱宝科技有限公司的糖原含量检测试剂盒(BC0345)来检测在不同浓度毛钩藤碱条件下的肝糖原含量。造模给药后将培养液换成无糖的葡萄糖输出缓冲液(无酚红，含2mM丙酮酸钠，20mM乳酸钠)培养细胞3h，再用葡萄糖氧化酶试剂盒检测缓冲液中葡萄糖含量来检测肝细胞葡萄糖输出量以对应糖异生程度。再通过实时定量PCR(qPCR)来检测HepG2细胞内糖异生相关的基因，如G6Pase，PEPCK，PGC-1α的表达量，细胞免疫荧光法检测细胞内葡萄糖转运体GLUT2的蛋白表达，seahorse技术检测细胞糖酵解能力，采用胞外产酸率，代表细胞的糖酵解水平。
实时定量PCR：为验证HepG2细胞内糖异生相关的基因的差异表达，使用实时定量PCR来检测G6Pase，PEPCK，PGC-1α的表达量。用TRIzol试剂提取HepG2细胞中的总mRNA并检测其浓度。用Biotool cDNA试剂盒将其逆转录为cDNA。操作中使用的水均为焦碳酸二乙酯(DEPC)处理后使用，以免RNA酶污染。Q-PCR反应条件：使用SYBR Green qPCR Mix试剂盒，扩增循环条件为95℃预变性10min，再经过40个循环的95℃变性30s，60℃30s退火以及72℃30s延伸。根据NCBI公布的G6Pase，PEPCK，PGC-1α的相关基因序列，应用Premier 5.0软件设计引物。以β-actin为内参基因。引物设计如下表1所示：
表1
细胞免疫荧光法：将24mm×24mm盖玻片浸泡，洗洁精清洗，自来水冲洗，稀盐酸浸泡8h，流水冲洗，铬酸浸泡过夜，流水冲洗，蒸馏水、三蒸水冲洗三次，置于玻璃培养皿中高压灭菌，烘干备用；在6孔细胞培养板中滴加少量细胞培养基，用无菌操作，夹取盖玻片，放入培养板孔中，使其贴附到孔底；取生长状态良好的HepG2细胞，PBS冲洗一遍，胰蛋白酶消化收集细胞，细胞计数，接种至6孔细胞培养板孔中的盖玻片上，4×105cells/ml，2ml/well，置于37℃，5％CO2细胞培养箱，培养24h。待HepG2细胞培养至长满单层，建立胰岛素抵抗模型并给药结束后，吸弃培养基，预冷的PBS冲洗3次，吸尽残留液体，4％多聚甲醛室温固定细胞，30min；弃去后用PBS洗涤3次；滴加3％BSA室温封闭2h；弃封闭液，将GLUT2(sc-518022，工作浓度1：100)一抗稀释液滴加至细胞表面，室温孵育2h；再4℃过夜，次日取出，复温，PBS洗涤3次；滴加FITC标记小鼠抗山羊IgG二抗(工作浓度1∶100)，37℃孵育2h；此过程注意避光，防止荧光基团的淬灭，PBS洗涤3次；DAPI染核，避光，加少许DAPI染液，覆盖住样品即可，室温孵育5min，弃去染液，PBS溶液洗涤3次；90％缓冲甘油封片，加防荧光淬灭液封片，尽量避免气泡，显微镜镜检并拍照记录。
seahorse技术：将HepG2细胞以8000个/孔种入八孔seahorse XFp细胞培养皿(每孔80微升培养基)，造模给药。提前往测量探针板中加入200微升校正液，随后将探针板放入37℃无CO2孵箱中过夜。药物作用结束后，弃去培养基，用seahorse生物能量分析培养基洗八孔板中的细胞3次，每次使用180微升，切勿将细胞洗脱落。随后将八孔板放入37℃无CO2敷箱中孵育1h。再将事先配好的浓度分别为10μM、5μM、10μM的寡霉素、羰基氰-对-三氟甲氧基本腙(FCCP)、抗霉素A和鱼藤酮工作液加入到探针板相对应的A、B、C孔中，然后将放上探针板的八孔板放入seahorse生物能量分析仪中同时检测氧耗量(OCR)和胞外产酸率(ECAR)。
2、实验结果：本实施例的毛钩藤碱对胰岛素抵抗型HepG2细胞内糖稳态的影响结果如图1、2所示，从图中可以看出，与对照组HGHI组相比较，毛钩藤碱可显著增加葡萄糖消耗和摄取量。肝糖原合成和肝糖输出试验结果如图3、4所示，从图中可以看出，毛钩藤碱可增加胰岛素抵抗的HepG2细胞中的糖原含量以及抑制其糖异生。糖异生相关的基因在胰岛素抵抗状态下表达的定量PCR如图5、6所示，从图中可以看出，对照组相关基因表达量异常升高，而毛钩藤碱可抑制其升高，表示毛钩藤碱可通过抑制糖异生达到维持糖稳态的作用。GLUT2作为一种在肝细胞中广泛存在的葡萄糖转运体，在胰岛素抵抗状态下表达受抑制，如图7所示，从图中可以看出，毛钩藤碱可增加GLUT2的表达，提高葡萄糖转运量。HepG2细胞的糖酵解量结果图8所示，从图中可以看出，毛钩藤碱可提高胰岛素抵抗HepG2细胞的糖酵解能力，从而改善糖代谢。
实施例3：毛钩藤碱在制备胰岛素抵抗相关药理途径中的作用通路
1、实验步骤：Western Blot检测Akt，PDK1，GSK3β的蛋白表达及磷酸化水平，以Akt，PDK1，GSK3β的磷酸化水平反应该通路的活性，再检测PI3K抑制剂LY294002对毛钩藤碱提高的胰岛素抵抗下HepG2细胞的葡萄糖摄取量，再通过Western Blot检测PI3K抑制剂LY294002显著抑制毛钩藤碱刺激下胰岛素抵抗HepG2细胞PI3K/Akt/GSK3β通路的激活，来反向验证毛钩藤碱通过激活胰岛素相关PI3K/Akt/GSK3β信号通路来提高葡萄糖摄取量。
Western Blot：使用RIPA蛋白裂解液提取培养细胞的总蛋白，BCA检测蛋白浓度，调整浓度后加入蛋白上样缓冲液煮沸变性。样品经SDS-PAGE后转移至NC膜。使用5％牛奶-TBST封闭液封闭1h，一抗4℃过夜后，用TBST洗膜三次，每次5min，再室温孵育二抗1h，洗膜后进行成像检测。
2、实验结果：Western Blot结果如图9所示，从图中可以看出，胰岛素抵抗可导致胰岛素信号通路PI3K/Akt/GSK3β中的关键蛋白Akt磷酸化障碍，而毛钩藤碱可激活该通路下的Akt、PDK1、GSK3β蛋白，使其磷酸化。如图10和11所示，从图中可以看出，加了PI3K/Akt/GSK3β通路的PI3K抑制剂LY294002后，葡萄糖摄取和PI3K/Akt/GSK3β通路的磷酸化均被抑制，说明毛钩藤碱改善胰岛素抵抗与激活PI3K/Akt/GSK3β通路有关。
实施例4：毛钩藤碱对db/db糖尿病鼠空腹血糖、改善其葡萄糖耐受以及肝脂质沉积试验
1、实验步骤：选取6组小鼠，每组12只。分为野生型C57BL/6小鼠(con)，组、db/db模型糖尿病鼠组、db/db+5mg/kg毛钩藤碱组、db/db+10mg/kg毛钩藤碱组、db/db+20mg/kg毛钩藤碱组、db/db+200mg/kg二甲双胍组。从6周龄开始灌胃给药，连续六周。每周测一次体重以及空腹血糖，最后一周做胰岛素耐受和口服葡萄糖耐受试验。小鼠的肝脏、胰脏、肾脏被取出，记录重量，用4％多聚甲醛溶液固定，按照以下步骤进行脱水：70％乙醇4h；80％乙醇2h；90％乙醇1h；95％乙醇2次，每次2h；100％乙醇2次，每次1h；二甲苯3次，每次1h；硬蜡2次，每次2h；脱水后石蜡包埋，切片(4μm厚)即可。再做后续的H&E染色或油红O染色试验。
H&E染色：将石蜡切片置于烘片机上60-65℃15min，按以下步骤常规脱蜡染色：二甲苯3次，每次7min；100％乙醇2次，每次5min；95％乙醇2次，每次5min；80％乙醇5min，70％乙醇5min，流水2min；苏木素染色，将载有玻片的染色架置于蒸馏水I--蒸馏水II--沥干--苏木素2min--大量自来水洗--静置于自来水中反蓝30min左右--显微镜观察，再脱水：50％乙醇--70％乙醇--80％乙醇--95％乙醇(每道0.5～1min)；伊红染色2min，然后脱水和透明，即80％乙醇15s；95％乙醇2次，每次1min；100％乙醇2次，每次1min；二甲苯3次，每次3min；中性树胶封片，期间避免气泡产生。
油红O染色：将进行石蜡包埋切片后的肝脏组织，进行OCT包埋，经冷冻切片机切片(4μm厚)，置于-80℃冰箱储藏。取出-80℃下的冰冻切片，复温干燥10min，50％乙醇稍洗，浸入60％异丙醇溶解的油红O染液10min，60％乙醇分化10s，流水冲洗2min，苏木素复染2s，1％盐酸乙醇分化15s，流水冲洗5min，甘油明胶封片，避免气泡产生。
2、实验结果：如图12所示，从图中可以看出，毛钩藤碱对db/db糖尿病鼠的体重没有明显影响。在不明显影响体重的前提下，毛钩藤碱可改善db/db糖尿病鼠的胰岛素抵抗状态，如图13，14，15所示，从图中可以看出，毛钩藤碱显著降低了db/db糖尿病鼠的空腹血糖以及口服葡萄糖耐受、胰岛素耐受量。如图16，17所示，从图中可以看出，毛钩藤碱可逆转db/db糖尿病鼠因脂肪肝造成的肝脏肥大等问题，并可改善糖尿病状态下偏淡黄的肝脏颜色，使其接近正常肝的褐红色。肝脏H&E、油红O和NAS评分结果如图18，19，20所示，从图中可以看出，毛钩藤碱可显著减少db/db糖尿病鼠肝中的脂质沉积，减少大泡样和小泡样脂变，恢复肝脏细胞的正常排列和形状，减少炎症细胞的浸润，减少非酒精性脂肪肝的活动度，表示毛钩藤碱可靶向到肝脏，改善糖尿病合并脂肪肝。胰脏的H&E结果如图21所示，从图中可以看出，毛钩藤碱可将db/db糖尿病鼠不规则的胰岛细胞形态以及空泡样变性改善到接近正常水平。肾脏的结果如图22所示，从图中可以看出，糖尿病早期的肾小球肥大增生，肾小球系膜基膜增厚，而毛钩藤碱可改善这一病变，使肾小球形状及体积趋于正常，减少肾小球系膜基膜增厚，这也说明毛钩藤碱可一定程度上减轻糖尿病肾病症状。
综上所述，本发明提供的毛钩藤碱对胰岛素抵抗、糖尿病及其并发症具有一定的治疗作用，且低浓度就可以起到疗效，并且可以逆转肝脏脂肪变性，同时，通过上述实验可以得知，毛钩藤碱治疗胰岛素抵抗、糖尿病及其并发症的药理作用途径通过激活疾病状态下受抑制的PI3K/Akt/GSK3β通路，来提高葡萄糖摄取量。为临床治疗II型糖尿病提供了新的治疗方法和治疗药物。
以上所述仅为本发明的实施例，并非因此限制本发明的专利范围，凡是利用本发明说明书及附图内容所作的等同变换，或直接或间接运用在相关的技术领域，均同理包括在本发明的专利保护范围内。
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<110> 澳门科技大学
<120> 毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用
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Claims (10)
1.毛钩藤碱在制备用于改善胰岛素抵抗、糖尿病及其并发症药物中的应用。
2.根据权利要求1所述的应用，其特征在于，所述毛钩藤碱由钩藤提取得到。
3.根据权利要求1所述的应用，其特征在于，所述糖尿病为II型糖尿病。
4.根据权利要求1所述的应用，其特征在于，所述并发症包括脂代谢紊乱疾病和/或肾病；优选地，所述脂代谢紊乱疾病为非酒精性脂肪肝。
5.根据权利要求1所述的应用，其特征在于，所述治疗胰岛素抵抗、糖尿病及其并发症药物为抑制胰岛素抵抗状态下的细胞的糖异生相关基因G6Pase、PEPCK或PGC-1α表达的药物。
6.根据权利要求1至5任一项所述的应用，其特征在于，所述药物还包括药学上可接受的载体或辅料。
7.根据权利要求1至5任一项所述的应用，其特征在于，所述药物的剂型为固体、半固体或液体的形式；优选为片剂、胶囊剂、软胶囊剂、颗粒剂、丸剂、口服液、干混悬剂、滴丸剂、干浸膏剂、注射剂或输注剂。
8.根据权利要求1至5任一项所述的应用，其特征在于，所述药物的给药方式为注射给药或口服给药；优选地，所述注射给药为静脉注射、肌肉注射、腹腔注射、皮内注射或皮下注射。
9.毛钩藤碱在制备PI3K/Akt/GSK3β信号通路激活剂中的应用。
10.根据权利要求9所述的应用，其特征在于，所述激活剂用于激活Akt、PDK1和GSK3β蛋白表达。
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6346280B1 (en) * | 1997-05-15 | 2002-02-12 | University Of Washington | Composition and methods for inhibiting the formation of brain amyloid deposits |
-
2020
- 2020-11-19 CN CN202011306445.4A patent/CN112494487A/zh active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6346280B1 (en) * | 1997-05-15 | 2002-02-12 | University Of Washington | Composition and methods for inhibiting the formation of brain amyloid deposits |
Non-Patent Citations (3)
| Title |
|---|
| ZHI-JUN WANG: "《A Novel Rhynchophylline Analog, Y396, Inhibits Endothelial Dysfunction Induced by Oxidative Stress in Diabetes Through Epidermal Growth Factor Receptor》", 《ANTIOXIDANTS & REDOX SIGNALING》 * |
| ZHI-JUN WANG: "《A Novel Rhynchophylline Analog, Y396, Inhibits Endothelial Dysfunction Induced by Oxidative Stress in Diabetes Through Epidermal Growth Factor Receptor》", 《ANTIOXIDANTS & REDOX SIGNALING》, vol. 32, no. 11, 4 March 2020 (2020-03-04), pages 743 - 765 * |
| 岳欣欣: "《糖尿病诊治策略》", 30 November 2018, pages: 21 - 22 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2024056498A1 (en) * | 2022-09-12 | 2024-03-21 | Société des Produits Nestlé S.A. | Oxindole alkaloid derivatives as inhibitors of sglt2 |
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