CN116554252A - 一种高结晶度别胆酸及其在制备预防和治疗胆汁淤积性肝病药物中的应用 - Google Patents
一种高结晶度别胆酸及其在制备预防和治疗胆汁淤积性肝病药物中的应用 Download PDFInfo
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Abstract
本发明属于医药领域,具体公开了一种高结晶度别胆酸及其在制备预防和治疗胆汁淤积性肝病药物中的应用,所述别胆酸在以2θ角表示的X‑射线粉末衍射图谱包括位于6.41±0.2°,6.74±0.2°,13.60±0.2°,14.84±0.2°,15.53±0.2°,17.13±0.2°,18.65±0.2°,22.51±0.2°处有特征峰。本发明发现别胆酸对3,5‑二乙氧基羰基‑1,4‑二氢‑2,4,6‑三甲基吡啶(DDC)诱导的小鼠胆汁淤积性肝纤维化具有明确的干预作用,缓解胆汁淤积造成的肝脏病理损伤、减轻肝纤维化程度、抑制胆管反应,可用于制备预防和治疗胆汁淤积性肝病的药物。
Description
技术领域
本发明属于医药领域,具体而言,涉及一种别胆酸的固体形态及其在治疗胆汁淤积性肝病中的应用。
背景技术
胆汁淤积是由于胆汁分泌过度及排泄障碍引起的一种病理生理过程,表现为肝脏以及体循环内胆酸、胆固醇及胆红素等胆汁成分的过度堆积,造成对肝细胞及机体的损伤。原发性胆汁性胆管炎(Primary biliary cholangitis,PBC)和原发性硬化性胆管炎(Primary Sclerosing Cholangitis,PSC)是成人最常见的慢性胆汁淤积性肝病,最终可能引起纤维化,肝硬化并最终导致肝功能衰竭。
目前为止,仅有熊去氧胆酸((Ursodeoxycholic Acid,UDCA)和奥贝胆酸(Obeticholic Acid,OCA)是美国食品药品监督管理局(FDA)批准的用于治疗胆汁淤积的药物。UDCA作为当前PBC的一线疗法,对每个阶段的患者都在一定程度上受益。然而,大约40%的PBC患者对UDCA的一线治疗反应不完全,并且存在疾病进展的风险(Lindor,Bowlus,Boyer,et al.Primary biliary cholangitis:2018practice guidance from theamerican association for the study of liver diseases.Hepatology,2019,69(1):394-419.)。OCA是一种FXR激动剂,2016年获得FDA批准上市作为PBC的二线治疗药物。OCA可以作为单药用于对UDCA不耐受的PBC患者,或者与UDCA联合使用,用于UDCA治疗一年后未能达到生化反应的PBC患者。但OCA会引起严重的瘙痒(Trauner,Nevens,Shiffman,etal.Long-term efficacy and safety of obeticholic acid for patients withprimary biliary cholangitis:3-year results of an international open-labelextension study.Lancet Gastroenterol Hepatol,2019,4(6):445-453.)。对于PSC,尽管已经有许多针对PSC的临床试验,但尚无有效的治疗方法可以延缓其进展或改善无移植生存期。因此,开发新的胆汁淤积性肝损伤治疗药物十分必要。
别胆酸(Allocholic acid,ACA),即3α,7α,12α-三羟基-5α-胆酸,是一种在脊椎动物中发现的胆汁酸,广泛分布于低等动物和一些哺乳动物中。CAS号为2464-18-8,化学结构如下所示:
别胆酸与传统的5β型存在着明显的结构特异性,两个六元环A和B采用稳定的椅式构象,但是近似存在于一个平面内。有研究表明,ACA可以作为阿尔茨海默症和不稳定型心绞痛的潜在生物标志物之一(Shao,Ouyang,Li,et al.Alteration of metabolic profileand potential biomarkers in the plasma of alzheimer's disease.Aging Dis,2020,11(6):1459-1470;Yu,Sun,Wang,et al.Biomarkers of unstable angina pectoris andyangxin decoction intervention:An exploratory metabonomics study of bloodplasma.Medicine,2017,96(21):e6998.)。还有研究称牛磺别胆酸(TACA)能刺激胆汁流量增加(Mendoza,Monte,Serrano,et al.Physiological characteristics of allo-cholicacid.J Lipid Res,2003,44(1):84-92.)。但是关于ACA在治疗胆汁淤积性肝病中的应用,目前还未见报道。
发明内容
本发明的目的之一是提供了一种晶型纯度高的别胆酸固体形态;
本发明的另一目的是提供了该晶型的别胆酸在预防和治疗胆汁淤积性肝病中的作用,确定了别胆酸对胆汁淤积性肝病的保护作用。
为实现上述目的,本发明的技术方案如下:
一种高结晶度别胆酸,在以2θ角表示的X-射线粉末衍射图谱包括位于6.41±0.2°,6.74±0.2°,13.60±0.2°,14.84±0.2°,15.53±0.2°,17.13±0.2°,18.65±0.2°,22.51±0.2°处有特征峰。
所述别胆酸是将别胆酸通过碱化溶于水,过滤,加酸缓慢结晶得到的。
所述别胆酸的制备方法包括如下步骤:
(1)将别胆酸粗粉加入到水中,然后加入氢氧化钠至基本澄清获得别胆酸粗品溶液;
(2)将别胆酸粗品溶液过滤后缓慢滴加酸性溶液至pH=2-5,得到别胆酸固体混合液;
(3)将别胆酸固体混合液搅拌2-12小时,析晶,过滤,得到高结晶度别胆酸。
优选地,所述酸性溶液为盐酸、硫酸、醋酸、柠檬酸等无机酸和低碳脂肪酸中的一种或两种以上。
高结晶度别胆酸在制备预防和治疗胆汁淤积性肝病药物中的应用。
所述胆汁淤积性肝病是原发性硬化性胆管炎。
所述胆汁淤积性肝病是原发性胆汁性胆管炎。
所述肝病为胆汁淤积引起的肝损伤和/或肝纤维化。
本发明优点在于:本发明通过3,5二乙氧基羰基1,4二氢2,4,6三甲基吡啶(DDC)诱导的小鼠胆汁淤积性肝纤维化模型,研究并证实了图1所示固体形态的别胆酸可以用于胆汁淤积性肝病的治疗。别胆酸能显著改善DDC诱导的胆汁淤积性肝纤维化小鼠的血清生化指标、缓解胆汁淤积造成的肝脏病理损伤、减轻肝纤维化程度、抑制胆管反应,可用于制备预防和治疗胆汁淤积性肝病的药物。
附图说明
图1:别胆酸固体的XRD图谱。
图2:别胆酸对DDC诱导的胆汁淤积小鼠血清生化指标的影响。(A)各组动物血清中天冬氨酸氨基转移酶(AST)的含量;(B)各组动物血清中丙氨酸氨基转移酶(ALT)的含量;(C)各组动物血清中碱性磷酸酶(ALP)的含量;(D)各组动物中血清中总胆汁酸(TBA)的含量。
图3:别胆酸对DDC诱导的胆汁淤积小鼠肝脏病理形态的影响。
图4:别胆酸对DDC诱导的胆汁淤积小鼠肝纤维化的影响。(A)天狼星红染色代表性图片;(B)半定量分析纤维化面积。
图5:肝纤维化的标志物α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白(COL1A1)的mRNA表达量分析。
图6:胆管上皮细胞增殖标志物细胞角蛋白19(CK-19)的mRNA表达量分析。
具体实施方式
为了更充分的理解本发明的技术内容,下面结合附图和具体实施例,进一步阐述本发明。这些实施例应理解为仅用于说明本发明,而不用于限制本发明的保护范围。在阅读了本发明记载的内容之后,本领域技术人员可以对本发明做各种改动或修改,这些等效变化和修改同样落入本发明权利要求所限定的范围。
实施例1
一种高结晶度别胆酸的制备:将5.1g别胆酸粗品加入到50mL水中,搅拌下缓慢滴加1N的NaOH溶液至近澄清,搅拌30分钟后过滤,除去少量的不溶物,得到别胆酸粗品溶液;然后滴加1N的盐酸溶液至pH为2.5,此时再滴加盐酸溶液也无沉淀析出,将此固液混合物搅拌4小时析晶,然后过滤,水洗,得到别胆酸固体3.8g,收率74.5%。
检测仪器及方法:
X-射线粉末衍射(XRD):仪器为Empyrean,采用铜靶波长为1.54nm的KαX-射线,在40kV和40mA的操作条件下进行。样品在室温下测试,将样品放在无反射板上,检测角度2θ范围为3-50°,步长为0.013°。
检测结果:
得到的别胆酸固体的X-射线粉末衍射图谱如图1所示,在下述2θ角有特征峰:6.41±0.2°,6.74±0.2°,13.60±0.2°,14.84±0.2°,15.53±0.2°,17.13±0.2°,18.65±0.2°,22.51±0.2°。
实施例2
ACA减轻DDC诱导的胆汁淤积性肝病小鼠的肝损伤
实验动物
54只6周龄雄性C57BL/6小鼠购自湖南斯莱克景达实验动物有限公司。所有实验动物均养在华南理工大学医学院实验动物中心SPF级屏障环境中,温度24-26℃,湿度40-70%,12h光/暗循环控制,所有实验动物均可自由获取食物和水。实验方案通过华南理工大学实验动物伦理委员会批准。
实验方案
54只雄性C57BL/6小鼠适应性饲养1周后将随机分为对照组9只和造模组45只,对照组(Control)给予常规小鼠饲料,造模组给予含0.025%DDC的饲料,共计9周。第2周起,将0.025%DDC的饲料造模小鼠随机分为模型组(Model),阳性药OCA组(10mg/kg),ACA低剂量组(3mg/kg),ACA中剂量组(10mg/kg),ACA高剂量组(50mg/kg),每组9只小鼠。从第2周起每天1次给予相应药物灌胃,连续给药8周,对照组和模型组给予以溶媒(5% HS15+5%丙二醇+90%水)灌胃,按照10ml/kg体积灌胃给药。给药8周后禁食过夜不禁水,随后眼眶采血,收集血液和肝脏等样品。血液样品室温静置分层后3500rpm,4℃离心15min,取上清存于于-80℃冰箱。摘取肝组织、观察其大体,并进行称重,其中,肝大叶浸泡于4%的多聚甲醛中固定,用于组织病理学检查,其余肝脏组织经液氮速冻后转移至-80℃保存备用。
统计学方法
所有数据均采用电子表格EXCEL进行处理,运用Graphpad Prism 8.0.1统计软件进行数据分析和作图,结果以“均值±标准误”(mean±SEM)的形式表示。多组间比较采用单因素方差分析(ANOVA)进行分析,符号说明:#是与正常组(Control)比较,#P<0.05,##P<0.01,###P<0.001,####P<0.0001;*是与模型组(Model)比较,*P<0.05,**P<0.01,***P<0.001,****P<0.0001。
表1ACA对DDC诱导的胆汁淤积小鼠体重,肝重及肝脏系数的影响
####P<0.0001,与Control组比较;*P<0.05,**P<0.01与Model组比较
实验结果
表1结果显示,Control组,Model组,OCA-10组,ACA-3组,ACA-10组和ACA-50组6组小鼠处死时体重均匀,无显著差异。用DDC造模9周后,Model组小鼠肝重显著高于Control组(p<0.0001),且肝脏系数也显著高于Control组(p<0.0001)。用10mg/kg和50mg/kg的ACA治疗后,能显著改善DDC造模引起的肝肿大(P<0.05),并明显降低肝脏系数。表明ACA对DDC诱导的胆汁淤积性肝损伤模型小鼠明显的治疗作用,可显著降低小鼠肝重和肝体比。
图2A和2B结果显示,肝损伤的重要指标AST和ALT在Model组小鼠中显著升高,说明用DDC造模9周后引起了严重的肝损伤。不同剂量的ACA和阳性药OCA均能显著降低血清ALT,50mg/kg的ACA显著降低血清AST((P<0.05)。血清ALP和TBA是胆汁淤积的重要指标。从图2C和2D中可以看出造模后血清ALP和TBA显著升高,说明造模引起了严重的胆汁淤积。10mg/kg和50mg/kg的ACA能显著降低血清ALP,50mg/kg的ACA能显著降低血清TBA(P<0.01)。从以上结果中看出50mg/kg的ACA对0.025% DDC造模引起的血清AST、ALT、ALP和TBA的升高均有一定的改善效果,说明ACA有治疗胆汁淤积的能力。
将小鼠肝脏固定、切片并进行苏木精和伊红(H&E)染色,光学显微镜下观察组织病变情况。如图3所示,Control组小鼠肝小叶结构正常,肝细胞形态正常,排列规则,无变性坏死现象,汇管区、肝小叶、中央静脉结构清晰;与Control组相比,DDC造模9周后,模型组小鼠肝脏汇管区可见明显的卟啉结晶沉积、炎性细胞浸润和不规则的新生胆管的出现。与Model组相比,OCA和ACA干预8周后胆汁淤积小鼠肝脏损伤现象得到了明显改善,其中以50mg/kg的ACA给药8周时改善更为明显,小鼠肝脏汇管区炎性细胞浸润面积减少,卟啉沉积改善、同时胆管增生现象亦得到了明显的抑制。上述结果表明,DDC可引起正常小鼠肝脏发生病理生理改变,其中以明显的卟啉结晶沉积、炎性细胞浸润和不规则的新生胆管的出现最为明显;ACA可以改善胆汁淤积小鼠肝脏病理生理情况。
实施例3
ACA减轻DDC诱导的胆汁淤积性肝病小鼠的肝纤维化
图4A是各组小鼠天狼星红染色代表性图片。与Control组相比,DDC诱导9周后模型组小鼠肝脏汇管区出现明显的胶原沉积;与Model组相比,ACA干预8周后一定程度上减轻了模型小鼠胶原沉积,且呈现剂量依赖性。阳性药OCA也显示出一定的抗肝纤维化效果。图4B是天狼猩红染色胶原阳性面积半定量结果,与Control组相比,DDC诱导9周后模型组小鼠肝脏胶原面积显著升高(p<0.0001);而与Model组相比,10mg/kg和50mg/kg的ACA及OCA均能显著减少小鼠肝组织胶原面积(p<0.0001)。
α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原蛋白(COL1A1)是体内纤维化的标志物。图5是α-SMA和COL1A1的qPCR结果,与Control相比,Model组COL1A1及α-SMA显著升高(分别为p<0.0001和p<0.05),ACA给药8周可明显抑制胆汁淤积小鼠肝组织COL1A1表达(p<0.01),对α-SMA也有降低趋势,但是无显著性。上述研究结果表明,ACA具有抑制胆汁淤积小鼠肝纤维化作用。
实施例4
ACA减轻DDC诱导的胆汁淤积性肝病小鼠的肝组织胆管反应
细胞角蛋白19(CK-19)是胆管上皮细胞的增殖标志物。图6是CK-19的qPCR结果,从图中可以看出与Control组相比,DDC诱导的胆汁淤积小鼠肝组织CK-19表达显著升高(P<0.001),与Model组相比,50mg/kg的ACA干预8周之后肝脏中CK-19表达明显降低(P<0.01),说明ACA能改善胆汁淤积小鼠肝脏汇管区胆管反应。
Claims (8)
1.一种高结晶度别胆酸,其特征在于,所述别胆酸在以2θ角表示的X-射线粉末衍射图谱包括位于6.41±0.2°,6.74±0.2°,13.60±0.2°,14.84±0.2°,15.53±0.2°,17.13±0.2°,18.65±0.2°,22.51±0.2°处有特征峰。
2.根据权利要求1所述的高结晶度别胆酸,其特征在于,所述别胆酸是将别胆酸通过碱化溶于水,过滤,加酸缓慢结晶得到的。
3.根据权利要求2所述的高结晶度别胆酸,其特征在于,所述别胆酸的制备方法包括如下步骤:
(1)将别胆酸粗粉加入到水中,然后加入氢氧化钠至基本澄清获得别胆酸粗品溶液;
(2)将别胆酸粗品溶液过滤后缓慢滴加酸性溶液至pH=2-5,得到别胆酸固体混合液;
(3)将别胆酸固体混合液搅拌2-12小时,析晶,过滤,得到高结晶度别胆酸。
4.根据权利要求3所述的高结晶度别胆酸,其特征在于,所述酸性溶液为盐酸、硫酸、醋酸、柠檬酸等无机酸和低碳脂肪酸中的一种或两种以上。
5.高结晶度别胆酸在制备预防和治疗胆汁淤积性肝病药物中的应用。
6.根据权利要求5所述的应用,其特征在于,所述胆汁淤积性肝病是原发性硬化性胆管炎。
7.根据权利要求5所述的应用,其特征在于,所述胆汁淤积性肝病是原发性胆汁性胆管炎。
8.根据权利要求5所述的应用,其特征在于,所述肝病为胆汁淤积引起的肝损伤和/或肝纤维化。
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Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106749473A (zh) * | 2017-01-13 | 2017-05-31 | 常德云港生物科技有限公司 | 一种从鸭胆汁中提取鹅去氧胆酸和别胆酸的方法 |
CN107973834A (zh) * | 2016-10-22 | 2018-05-01 | 合帕吉恩治疗公司 | 作为fxr/tgr5调节剂的胆酸衍生物 |
CN108348543A (zh) * | 2015-09-21 | 2018-07-31 | 英特塞普特医药品公司 | 促进肝再生的方法 |
CN108904537A (zh) * | 2018-08-02 | 2018-11-30 | 上海凯宝药业股份有限公司 | 一种家禽胆转化物在制备抗胆汁淤积性肝病药物中的应用 |
CN109810159A (zh) * | 2019-01-22 | 2019-05-28 | 常德云港生物科技有限公司 | 一种能从鸭胆汁中提高别胆酸收率的方法 |
CN109929005A (zh) * | 2017-12-19 | 2019-06-25 | 西安奥立泰医药科技有限公司 | 用于代谢性疾病治疗的化合物及其制备方法和应用 |
CN111116699A (zh) * | 2015-04-28 | 2020-05-08 | 江苏豪森药业集团有限公司 | 胆酸衍生物及其制备方法和医药用途 |
CN112220792A (zh) * | 2020-09-30 | 2021-01-15 | 浙江大学 | 石胆酸用于制备缓解肝纤维化的药物中的应用 |
CN112409435A (zh) * | 2019-08-23 | 2021-02-26 | 深圳云合医药科技合伙企业(有限合伙) | 胆汁酸衍生物及其组合物和应用 |
CN114144185A (zh) * | 2019-05-30 | 2022-03-04 | 英特塞普特医药品公司 | 用于治疗胆汁淤积性肝病的包含fxr激动剂和贝特类的药物组合物 |
-
2023
- 2023-04-07 CN CN202310365309.XA patent/CN116554252A/zh active Pending
Patent Citations (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111116699A (zh) * | 2015-04-28 | 2020-05-08 | 江苏豪森药业集团有限公司 | 胆酸衍生物及其制备方法和医药用途 |
CN108348543A (zh) * | 2015-09-21 | 2018-07-31 | 英特塞普特医药品公司 | 促进肝再生的方法 |
CN107973834A (zh) * | 2016-10-22 | 2018-05-01 | 合帕吉恩治疗公司 | 作为fxr/tgr5调节剂的胆酸衍生物 |
CN106749473A (zh) * | 2017-01-13 | 2017-05-31 | 常德云港生物科技有限公司 | 一种从鸭胆汁中提取鹅去氧胆酸和别胆酸的方法 |
CN109929005A (zh) * | 2017-12-19 | 2019-06-25 | 西安奥立泰医药科技有限公司 | 用于代谢性疾病治疗的化合物及其制备方法和应用 |
CN108904537A (zh) * | 2018-08-02 | 2018-11-30 | 上海凯宝药业股份有限公司 | 一种家禽胆转化物在制备抗胆汁淤积性肝病药物中的应用 |
CN109810159A (zh) * | 2019-01-22 | 2019-05-28 | 常德云港生物科技有限公司 | 一种能从鸭胆汁中提高别胆酸收率的方法 |
CN114144185A (zh) * | 2019-05-30 | 2022-03-04 | 英特塞普特医药品公司 | 用于治疗胆汁淤积性肝病的包含fxr激动剂和贝特类的药物组合物 |
CN112409435A (zh) * | 2019-08-23 | 2021-02-26 | 深圳云合医药科技合伙企业(有限合伙) | 胆汁酸衍生物及其组合物和应用 |
CN112220792A (zh) * | 2020-09-30 | 2021-01-15 | 浙江大学 | 石胆酸用于制备缓解肝纤维化的药物中的应用 |
Non-Patent Citations (2)
Title |
---|
XUE HAN ET AL.: "Allocholic acid protects against α-naphthylisothiocyanateinduced cholestasis in mice by ameliorating disordered bile acid homeostasis", 《J APPL TOXICOL.》, 15 November 2023 (2023-11-15), pages 1 - 13 * |
韩平等: "肝内胆汁淤积发病机制及药物治疗新进展", 《胃肠病学和肝病学杂志》, vol. 25, 31 May 2016 (2016-05-31), pages 584 - 588 * |
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