CN107970233A - 磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用 - Google Patents
磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用 Download PDFInfo
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Abstract
本发明公开了磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用,提供了神经氨酸酶与心肌缺血损伤的关联性,证明通过抑制神经氨酸酶的活性可以缓解心肌缺血损伤,神经氨酸酶可以作为筛选预防、缓解和/或治疗心肌缺血损伤药物的靶标;磷酸奥司他韦作为神经氨酸酶抑制剂可以有效缓解心肌缺血引起的心肌损伤。
Description
本发明是2016年03月22日递交的申请号为“2016101662535”、发明名称为“神经氨酸酶及其抑制剂在心肌缺血及心肌梗死中的应用”的专利申请的分案申请。
技术领域
本发明属于生物医药领域,涉及已知化合物的新用途,具体涉及磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用。
背景技术
心血管疾病是威胁人类生命健康的重大疾病,随着社会进步和人民生活水平提高,其发病率逐年增长,2014年因心血管疾病死亡人数约占全球总死亡人数的30%,而心肌缺血性疾病又是心血管疾病中的焦点,并可发展为心律失常、心肌梗死,常可危及生命。因而如何切实有效地减轻因心肌缺血引起的损伤已成为当今医药界面临的一个热点问题。
心肌缺血损伤是由于心肌细胞供氧不足而造成的心肌细胞坏死或功能受损,目前临床常用药物多是通过改善细胞能量代谢、抑制炎症反应、保护血管、减轻心肌细胞钙超载等机制治疗心肌缺血,常见的包括硝酸脂类药物如硝酸甘油,硝酸异山梨酯;β-受体阻断剂如普萘洛尔;钙通道阻断剂如硝苯地平,维拉帕米以及抗血小板和抗血栓形成药如双嘧达莫等。
磷酸奥司他韦是一种公认有效的抗流感药物,目前未见其用于治疗心肌缺血损伤的报道。
发明内容
本发明的目的在于提供磷酸奥司他韦在制备治疗心肌缺血及心肌梗死的药物中的应用。
本发明的上述目的是通过下面的技术方案得以实现的:
磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用。
进一步地,所述心肌缺血为心肌梗死引发的心肌缺血。
本发明发现磷酸奥司他韦可以有效改善心肌缺血引起的心肌损伤,可以用于制备治疗心肌缺血损伤的药物。
附图说明
图1A为空白对照组大鼠心电图;
图1B为异丙肾上腺素造模后模型组大鼠心电图;
图2A为异丙肾上腺素造模后扎那米韦低剂量组大鼠心电图;
图2B为异丙肾上腺素造模后扎那米韦高剂量组大鼠心电图;
图3A为异丙肾上腺素造模后磷酸奥司他韦低剂量组大鼠心电图;
图3B为异丙肾上腺素造模后磷酸奥司他韦高剂量组大鼠心电图;
图4A为空白对照组大鼠心脏病理切片电镜图;
图4B为异丙肾上腺素造模后模型组大鼠心脏病理切片电镜图;
图5A为异丙肾上腺素造模后扎那米韦低剂量组大鼠心脏病理切片电镜图;
图5B为异丙肾上腺素造模后扎那米韦高剂量组大鼠心脏病理切片电镜图;
图6A为异丙肾上腺素造模后磷酸奥司他韦低剂量组大鼠心脏病理切片电镜图;
图6B为异丙肾上腺素造模后磷酸奥司他韦高剂量组大鼠心脏病理切片电镜图;
图7为异丙肾上腺素造模后各组心肌神经氨酸酶表达水平的影响。
具体实施方式
下面结合实施例进一步说明本发明的实质性内容,但并不以此限定本发明保护范围。尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。本发明中,未做详细描述或特别强调的试验材料或试验方法均为本领域常规试验材料或试验方法,本领域普通技术人员可以获得该试验材料或具备开展该试验的能力。
一、试验材料
1、仪器和试剂
生理记录仪:BL-420S生理机能实验系统(中国成都泰盟)、动物呼吸机HX-300S(中国成都泰盟)、MP120-1电子秤(上海第二天平仪器厂)、动物手术器械等。
盐酸异丙肾上腺素(ISO)购自上海阿拉丁生化科技股份有限公司,纯度>99%、0.9%生理盐水购自国药集团、扎那米韦(ZA)购自大连美仑生物技术有限公司,纯度>98%、磷酸奥司他韦(OS)购自大连美仑生物技术有限公司,纯度>98%、3%水合氯醛。
2、试验动物
SD雄性大鼠120只,购自上海西普尔-必凯公司。
二、试验方法
1、ISO造大鼠急性心肌缺血模型:选取SD大鼠,根据体重分为空白对照组、模型组、ZA高剂量组(0.5mg/kg i.v.)、ZA低剂量组(0.2mg/kg i.v.)、OS高剂量组(10mg/kg p.o.)、OS低剂量组(5mg/kg p.o.)六组,每组10只。给药组连续给药3天,第二天给药30min后皮下注射ISO,剂量为60mg/kg,第三天同样给药30min后皮下注射ISO,剂量为60mg/kg,第4天收集血液、心脏等组织样本。
2、左冠状动脉前降支结扎造大鼠急性心肌缺血模型:选取SD大鼠,根据体重分为空白对照组、模型组、ZA高剂量组(0.5mg/kg i.v.)、ZA低剂量组(0.2mg/kg i.v.)、OS高剂量组(10mg/kgp.o.)、OS低剂量组(5mg/kg p.o.)六组,每组10只。给药组预先24h和12h给药,而后将大鼠3%水合氯醛10ml/kg麻醉,仰卧固定于大鼠板上。于左侧4、5肋间切口撕开心包,暴露心脏,轻压胸廓挤出心脏,于肺动脉圆锥及左心房间找出冠状动脉左前降支,以0号线立即结扎冠状动脉左前降支根部(肺动脉圆锥与左心耳处),将心脏送回胸腔,并挤出胸腔内血液和气体,迅速关闭胸腔,缝合皮肤,开胸时间不超过30s。采用BL-420S生理机能实验系统对每只大鼠进行心电图的测定。造模后24h收集血液、心脏等组织样本:摘眼球取血,分离血清,-20℃保存待测;脱颈椎处死动物,迅速取出心脏,以冰生理盐水洗去残留血液,除去大血管、结缔组织,滤纸吸干,称全心重。取心尖部位至10%福尔马林溶液中固定,用于病理学检查,其余心脏减碎后,以10倍量Tris-HCl(pH7.4)缓冲液在冰浴下制成10%的心脏匀浆,于4℃,1000g离心10min,弃沉淀,取上清进行各项酶学指标的检测。
3、观察指标及方法
3.1记录心电图
采用标准Ⅱ导联记录各组大鼠心电图
3.2观察心脏病理切片
取心尖部位的心肌组织经10%甲醛溶液固定,常规取材,脱水,石蜡包埋,制片(4μm厚),HE染色,在光学显微镜下观察。
3.3心肌神经氨酸酶水平测定方法
使用ELISA法检测大鼠心肌神经氨酸酶水平,检测试剂盒为神经氨酸酶检测试剂盒,按照检测试剂盒的操作说明书进行测定。
3.4血清心肌损伤指标CK-MB和D-LDH检测方法
分别按照CK-MB检测试剂盒(罗氏)和D-LDH检测试剂盒(Invitrogen)操作说明书测定CK-MB和D-LDH。
三、试验结果
1、对心电图的影响
图1~图3所示的典型心电图记录结果表明:异丙肾上腺素造模后,模型组出现明显心肌损伤,神经氨酸酶抑制剂扎那米韦和磷酸奥司他韦能有效改善心肌损伤,并呈现剂量依赖性。
2、对心肌细胞形态的影响
图4~图6所示的典型心脏病理切片表明:异丙肾上腺素造模后,模型组出现明显心肌损伤,表现为心肌细胞形态不规则,细胞间裂隙明显,大量炎症细胞浸润,神经氨酸酶抑制剂扎那米韦和磷酸奥司他韦能有效改善心肌损伤,并呈现剂量依赖性。
3、对心肌神经氨酸酶表达水平的影响
从表1和图7可以看出,异丙肾上腺素造模后,ELISA法检测大鼠心肌细胞中神经氨酸酶表达,结果显示模型组心肌缺血后神经氨酸酶表达上调,神经氨酸酶抑制剂扎那米韦和磷酸奥司他韦能有效抑制心肌缺血引起的神经氨酸酶表达升高,并呈现剂量依赖性。
表1各组心肌神经氨酸酶表达水平
4、对心肌损伤指标在血清中表达水平的影响
CK-MB和D-LDH是心肌缺血损伤的血清标志物,异丙肾上腺素造急性心肌缺血模型后,血清CK-MB、LDH水平上升,给药组则显著性降低。结果见表2。
表2血清CK-MB、D-LDH表达水平
在大鼠冠状动脉左前降支结扎模型中,我们得到了与异丙肾上腺素造急性心肌缺血模型类似的结论。冠脉结扎模型组心肌神经氨酸酶活力升高,血清CK-MB、LDH水平上升,给药组则显著性降低。同时,扎那米韦与奥司他韦给药能显著改善大鼠心电图及心脏病理切片。
上述实施例的作用在于说明本发明的实质性内容,但并不以此限定本发明的保护范围。本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和保护范围。
Claims (2)
1.磷酸奥司他韦在制备治疗心肌缺血损伤的药物中的应用。
2.根据权利要求1所述的应用,其特征在于:所述心肌缺血为心肌梗死引发的心肌缺血。
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CN107890565B (zh) * | 2017-11-27 | 2020-02-07 | 中国药科大学 | 神经氨酸酶及抑制剂在制备治疗心肌纤维化和心室肥厚的药物中的应用 |
CN107715107B (zh) * | 2017-11-27 | 2020-03-06 | 中国药科大学 | 神经氨酸酶及抑制剂在制备治疗高血压的药物中的应用 |
CN109701022A (zh) * | 2019-02-22 | 2019-05-03 | 华中科技大学同济医学院附属同济医院 | 神经氨酸酶抑制剂在暴发性心肌炎药物中的应用 |
CN115590847A (zh) * | 2021-07-08 | 2023-01-13 | 中国医学科学院药物研究所(Cn) | 戈米辛d在制备治疗或预防心肌缺血性疾病药物中的应用 |
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