CN115282140B - Aki治疗剂及dmxaa在制备该治疗剂中的应用 - Google Patents
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Abstract
本发明涉及2,5‑己酮可可碱在制备急性肾损伤治疗剂中的应用,还涉及包含2,5‑己酮可可碱的急性肾损伤治疗剂,以及包含该急性肾损伤治疗剂的用于治疗实体瘤的药剂组。顺铂是肿瘤治疗中十分常用的化疗药剂,但是顺铂施用导致的并发症急性肾损伤对患者健康损害严重。本发明证实2,5‑己酮可可碱可治疗和缓解因顺铂施用导致的急性肾损伤,为化疗中缓解该并发症提供了潜在新药。通过本发明可预期,可在化疗中将2,5‑己酮可可碱与顺铂联用,从而在杀伤肿瘤的过程中减轻甚至消除急性肾损伤。
Description
技术领域
本发明涉及肿瘤及其并发症的治疗领域,更特别得,涉及2,5-己酮可可碱在制备急性肾损伤治疗剂中的应用,以及含有2,5-己酮可可碱的急性肾损伤治疗剂。
背景技术
顺铂是临床常用的广谱抗癌药物,对肺癌、鼻咽癌、卵巢癌等多种实体肿瘤均具有抑制作用。顺铂的肾脏转运受近端小管转运蛋白调节,在肾近端小管上皮细胞中积累导致细胞损伤甚至死亡。所以,接受顺铂化疗的患者中,有很多会表现出急性肾损伤(AKI)的并发症。
2,5-己酮可可碱(DMXAA,又名伐地美生),结构如式I所示,被发现可以特异性地破坏肿瘤血管,因此作为血管损伤剂被用于癌症治疗中。从现有研究来看,DMXAA还可以激活固有免疫系统、刺激炎症因子TNF、IL-6等和趋化因子MIP-1α、IP-10等的生成,进一步促进肿瘤特异性的炎症反应,因而可作为抑制肿瘤的潜在药物。此外,科研人员还发现,DMXAA可以激活STING(TMEM173)蛋白,因此被用作STING的激动剂。有研究表明,STING蛋白具有抗肿瘤的作用,但是,其介导的炎症与凋亡同样导致包括AKI在内的多种脏器损伤。并且,已有研究显示,STING的抑制剂C-176和H-151均可明确缓解顺铂诱导的AKI。因此,从现有的理论上而言,DMXAA的施用很可能会加剧顺铂诱导的AKI。
发明内容
我们在研究DMXAA的过程中,意外地发现,DMXAA的施用不仅没有加剧顺铂诱导的AKI,甚至对AKI具有显著的缓解和治疗作用。基于上述发现,本发明提供了2,5-己酮可可碱在制备急性肾损伤治疗剂中的应用。
在一个具体实施方案中,所述急性肾损伤为顺铂诱导的急性肾损伤。
本发明还提供了包含2,5-己酮可可碱的急性肾损伤治疗剂。
在一个具体实施方案中,所述急性肾损伤为顺铂诱导的急性肾损伤。
在一个具体实施方案中,所述急性肾损伤治疗剂的溶剂为10%的吐温-80。
本发明还提供了一种用于治疗实体瘤的药剂组合,包括顺铂和急性肾损伤治疗剂,所述急性肾损伤治疗剂为2,5-己酮可可碱。
顺铂是肿瘤治疗中十分常用的化疗药剂,但是顺铂施用导致的并发症AKI对患者健康损害严重。本发明证实DMXAA可治疗和缓解因顺铂施用导致的AKI,为化疗中缓解该并发症提供了潜在新药。通过本发明可预期,可在化疗中将DMXAA与顺铂联用,从而在杀伤肿瘤的过程中减轻甚至消除AKI。
附图说明
图1为三个组的小鼠血清BUN与Scr含量统计图。
图2为三个组的小鼠肾组织H&E染色照片(20倍)。
图3为三个组的小鼠肾组织TUNEL染色照片。
图4为三个组的小鼠肾组织中的KIM1与NGAL的免疫印迹染色照片。
图5为三个组的小鼠血清中TNF-α含量统计图。
具体实施方式
以下结合附图对本发明的原理和特征进行描述,所举实例只用于解释本发明,并非用于限定本发明的范围。
1.小鼠模型构建
采用7-8周龄的野生型雄性小鼠C57BL/6,分成三组,加药组(XAA+Cis)先进行腹腔注射DMXAA(10mg/kg),对照组(Saline)和模型组(Cis)给予等体积的10%的吐温-80。1h后,模型组和加药组给予顺铂(25mg/kg)腹腔注射,对照组给予等体积的0.9%生理盐水。
顺铂注射后,每隔24h加药组给予一次腹腔注射DMXAA(10mg/kg),相应的,对照组与模型组给予等体积10%的吐温-80。顺铂注射72小时后收取所有小鼠,采集血清与肾组织,用于后续研究。
2.肾功能指标的检测
对采集的小鼠血清,采用生化法检测血尿素氮(BUN)与血肌酐(Scr)的含量。结果如图1所示,模型组的血清中BUN与Scr的水平明显升高,而加药组的两个指标显著低于模型组,但是高于对照组。这说明,DMXAA缓解了顺铂导致的肾功能生化指标的损伤。
3.肾组织染色
取小鼠肾组织,经多聚甲醛溶液固定后进行脱水、石蜡包埋、切片,然后进行H&E染色,结果如图2所示,正常对照组肾组织刷状缘清晰明显,小管结构完整;模型组中的肾小管损伤严重,刷状缘部分丢失、不清晰,小管扩张且出现管型。而加药组肾组织的病理变化明显得到改善,小管结构基本清晰完整,接近正常组织,管型消失。提示DMXAA可显著缓解顺铂导致的肾组织病理损伤。
取小鼠肾组织石蜡切片,应用TUNEL染色试剂盒进行组织染色,检测肾小管细胞凋亡情况。结果如图3所示,模型组中显示出大量细胞凋亡的情况,而加药组中只有个别凋亡细胞。
上述实验结果说明,DMXAA明显缓解了顺铂导致的肾组织病理损伤,以及肾小管上皮细胞凋亡。
4.肾小管损伤标志物的表达检测
提取小鼠肾组织蛋白,使用免疫印迹(Western blot)法检测肾组织中小管损伤标记物KIM1和NGAL的蛋白表达情况。结果如图4所示,KIM1和NGAL在模型组中大量表达,而在对照组和加药组中基本不表达。
使用TNF-α的ELISA检测试剂盒,检测小鼠血清中TNF-α的含量。结果如图5所示,加药组小鼠血清中TNF-α含量虽然高于对照组,但是明显低于模型组。
以上实验结果说明,从分子水平上也体现出了DMXAA对顺铂诱导的肾损伤的缓解。
综合上述实验结果说明,DMXAA的注射,无论在生化功能水平、组织结构水平还是在分子标志物水平,都显示出对顺铂诱导的肾损伤的缓解。因此,可以认为,DMXAA对顺铂诱导的肾损伤具有治疗作用。此外,由于上述实验显示DMXAA与顺铂联合施用可缓解和治疗顺铂诱导的急性肾损伤,这也意味着,这两者可联合施用以治疗相应的肿瘤,因此本发明同样应当涵盖这种联合施用的药剂组合。
以上所述仅为本发明的较佳实施例,并不用以限制本发明,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (1)
1.2,5-己酮可可碱在制备急性肾损伤治疗剂中的应用,所述急性肾损伤为顺铂诱导的急性肾损伤。
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