CN116421594A - 一种nlrp3炎性小体抑制剂及其应用 - Google Patents
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Abstract
本发明提供了一种NLRP3炎性小体抑制剂，所述NLRP3炎性小体抑制剂为金属配合物二乙基二硫羰氨酸铜盐，并提供了NLRP3炎性小体抑制剂在制备治疗NLRP3炎性小体活化疾病药物中的应用，属于生物医药技术领域。本发明的NLRP3炎性小体抑制剂可直接抑制NLRP3炎性小体组装，达到抑制NLRP3炎性小体活化的效果，进而减少炎症因子IL‑1β的释放，最终可以起到抑制细胞焦亡的效果，明显减轻NLRP3炎性小体活化引起炎症风暴和器官损伤，为制备治疗NLRP3炎性小体活化疾病药物提供了全新的选择。
Description
技术领域
本发明涉及生物医药技术领域，具体涉及一种NLRP3炎性小体抑制剂及其应用。
背景技术
NLRP3炎性小体是由NLRP3、ASC和Caspase-1组装的一类蛋白复合体，且其活化需要NEK7参与，激活后的NLRP3炎性小体通过调节Caspase-1的活化促进IL-1β等因子的成熟和释放，继而介导细胞焦亡，诱发一系列炎症和免疫反应。众多研究表明：NLRP3炎性小体的异常活化与脓毒症、炎症性肠炎、非酒精性脂肪肝、中风、痛风、糖尿病、动脉粥样硬化、阿尔兹海默症、哮喘等发生发展密切相关，已对人类健康造成巨大威胁。以脓毒症为例，全球每年大约3100万的新发病例，约530万人因此失去生命，死亡率已超过前列腺癌、乳腺癌及艾滋病三种疾病的总和，是重症患者最为常见的死因。然而，迄今为止针对脓毒症的治疗手段仍十分有限，临床上主要依赖于抗生素的使用和器官功能支持，并且面临微生物耐药和医疗资源消耗大的困境。近年来发现NLRP3炎性小体诱导的细胞焦亡是脓毒症的重要病理表现，免疫细胞因短时间内发生大量焦亡被耗竭，引发强烈的细胞因子风暴和器官功能障碍，最终导致患者免疫失调继而死亡风险显著增加。因此，NLRP3炎性小体是脓毒症及上述多种疾病的重要治疗靶点。目前针对NLRP3炎性小体的抑制剂以小分子化合物为主，如MCC950、AZD9056、CE-224535和GSK1482169等，但大多数都因选择性差、毒副作用大、生物利用度低而无法实现临床转化。因此，开发高效低毒的NLRP3炎性小体特异性抑制剂迫在眉睫。
二乙基二硫羰氨酸铜盐是由二硫卡钠与铜离子反应而成的金属配合物。目前，二硫卡钠与铜离子反应形成的的金属配合物主要用于肿瘤的治疗药物的研究，尚无该金属配合物用于炎症因子抑制相关机制和效果的研究。
发明内容
为解决上述技术问题，本发明提出了一种NLRP3炎性小体抑制剂及其应用，其目的是提供一种为NLRP3炎性小体抑制提供新的治疗靶点和药物制备的选择，采用以下技术方案：一种NLRP3炎性小体抑制剂，所述NLRP3炎性小体抑制剂为二乙基二硫羰氨酸铜盐。
作为优选，所述二乙基二硫羰氨酸铜盐由二硫卡钠和铜离子反应形成。
作为优选，所述NLRP3炎性小体抑制剂可抑制NLRP3炎性小体的活化。
作为优选，所述NLRP3炎性小体抑制剂可抑制炎症因子IL-1β的释放。
作为优选，所述NLRP3炎性小体抑制剂可抑制细胞焦亡。
基于一个总的发明构思，所述的NLRP3炎性小体抑制剂在制备治疗NLRP3炎性小体活化疾病药物中的应用。
作为优选，所述NLRP3炎性小体活化疾病为脓毒症、炎症性肠炎、非酒精性脂肪肝、中风、痛风、糖尿病、动脉粥样硬化、阿尔兹海默症、哮喘中的一种。
作为优选，所述NLRP3炎性小体活化相关疾病药物治疗药物的剂型为注射剂、口服剂或外用剂。
本发明提供的发明方案的有益效果为：
1、本发明通过试验研究创造性的发现了一种全新的NLRP3炎性小体抑制剂：二乙基二硫羰氨酸铜盐，该炎性小体抑制剂可直接抑制NLRP3炎性小体组装，达到抑制NLRP3炎性小体活化的效果，进而减少炎症因子IL-1β的释放，最终可以起到抑制细胞焦亡的效果，明显减轻NLRP3炎性小体活化引起炎症风暴和器官损伤，为制备治疗NLRP3炎性小体活化疾病药物提供了全新的选择。
2、本发明提供的抑制剂可以抑制NLRP3炎性小体活化诱导的炎症因子释放及细胞焦亡，从而减少脓毒症小鼠肺泡塌陷、肺泡壁增厚以及炎症细胞浸润的病理损伤、降低小鼠血清NLRP3特异性炎症因子水平。该结果说明，该抑制剂二乙基二硫羰氨酸铜盐能够用于防治脓毒症等由炎性小体引起的相关疾病的治疗药物制备。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1是实验例1中抑制剂对由脂多糖+尼利亚菌素诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的细胞焦亡实验结果；
图2是实验例1抑制剂对由脂多糖+尼利亚菌素诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的炎症因子IL-1β释放实验结果；
图3是实验例1中抑制剂对由脂多糖+三磷酸腺苷诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的细胞焦亡实验结果；
图4是实验例1抑制剂对由脂多糖+三磷酸腺苷诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的炎症因子IL-1β释放实验结果；
图5是实验例1中抑制剂对由脂多糖+尿酸结晶诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的细胞焦亡实验结果；
图6是实验例1抑制剂对由脂多糖+尿酸结晶诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的炎症因子IL-1β释放实验结果；
图7是实验例1抑制剂对由脂多糖+尼利亚菌素诱导NLRP3炎性小体活化的小鼠原代腹腔巨噬细胞的NLRP3炎性小体组分的实验结果；
图8是实验例2抑制剂对脂多糖诱导的脓毒症小鼠血清中炎症因子IL-1β释放实验结果；
图9是实验例2抑制剂对由脂多糖诱导的脓毒症小鼠血清中炎症因子IL-6释放实验结果；
图10是实验例3抑制剂对脂多糖诱导的脓毒症小鼠肺部病理损伤的H&E染色结果。
具体实施方式
为使本发明要解决的技术问题、技术方案和优点更加清楚，下面将结合附图及具体实施例进行详细描述。
以下实施例用于说明本发明，但不用来限制本发明的范围。在不背离本发明精神和实质的情况下，对本发明方法、步骤或条件所作的修改或替换，均属于本发明的范围。
若未特别指明，实施例中所用的技术手段为本领域技术人员所熟知的常规手段；若未特别指明，实施例中所用试剂均为市售。
材料与试剂
二硫卡钠(Sigma)、氯化铜(麦克林)、脂多糖(Sigma)、LDH试剂盒(碧云天)、鼠IL-1βELISA试剂盒(eBioscience)、鼠IL-6ELISA试剂盒(eBioscience)。
实验细胞
小鼠原代腹腔巨噬细胞，由实验室人员提取。
实验动物
SPF级C57BL/6J雄性小鼠，采购于湖南斯莱克景达实验动物有限公司。
实施例1
金属配合物二乙基二硫羰氨酸铜盐的制备
用DMSO配制40mM二硫卡钠和20mM氯化铜溶液，将两溶液同体积混合可生成沉淀，即金属配合物二乙基二硫羰氨酸铜盐，经抽滤、洗涤、冷冻干燥可得NLRP3炎性小体抑制剂二乙基二硫羰氨酸铜盐粉末。
实验例1
抑制NLRP3炎性小体活化的作用
向小鼠腹腔注入2mL含3％脱水硫羟乙酸的培养基，3天后收集腹腔细胞。细胞悬液以1000rpm离心5分钟，用含有10％FBS的RPMI 1640重悬。细胞悬液置入培养板，37℃培养2h后，吸弃上清，用预热培养基洗1次，留下贴壁细胞，即得新鲜分离的小鼠原代腹腔巨噬细胞。
细胞铺板过夜后，设置对照组、脂多糖+尼日利亚菌素模型组、脂多糖+三磷酸腺苷模型组、脂多糖+尿酸结晶模型组、金属配合物0.5μM组、金属配合物1μM组、金属配合物2μM组。脂多糖+尼日利亚菌素模型组、脂多糖+三磷酸腺苷模型组、脂多糖+尿酸结晶模型组、金属配合物0.5μM组、金属配合物1μM组、金属配合物2μM组予以100ng/mL脂多糖预处理3h后，分别在金属配合物0.5μM组、金属配合物1μM组、金属配合物2μM组予以实施例1制备的相对应浓度的金属配合物二乙基二硫羰氨酸铜盐孵育1h，其余两组予以同体积溶剂。而后，脂多糖+尼日利亚菌素模型组、金属配合物0.5μM组、金属配合物1μM组、金属配合物2μM组予以10μM的尼日利亚菌素活化NLRP3炎症小体，1h后收集细胞上清液，用LDH试剂盒检测LDH的释放量反应细胞焦亡水平，ELISA法检测细胞炎症因子IL-1β的释放以及收集细胞，WesternBlotting检测NLRP3炎性小体组分表达。
结果如图1-图7所示，由图1和图2可知，脂多糖+尼日利亚菌素可以激活NLRP3炎性小体，导致细胞焦亡，细胞膜破裂，释放出大量LDH和炎症因子IL-1β，予以不同浓度的金属配合物二乙基二硫羰氨酸铜盐治疗后，LDH和IL-1β释放显著减少，并呈浓度依赖的趋势，图1说明金属配合物二乙基二硫羰氨酸铜盐可以明显抑制细胞焦亡，图2说明金属配合物二乙基二硫羰氨酸铜盐可以显著减少炎症因子IL-1β的释放；而同时在图3和图4，以及图5和图6中，我们发现脂多糖+三磷酸腺苷或者脂多糖+尿酸结晶同样可以激活NLRP3炎性小体，从而导致细胞焦亡，图3和图4说明金属配合物二乙基二硫羰氨酸铜盐可以明显抑制由脂多糖+三磷酸腺苷诱导的细胞焦亡和IL-1β的释放；图5和图6说明金属配合物二乙基二硫羰氨酸铜盐可以显著减少脂多糖+尿酸结晶诱导的细胞焦亡和IL-1β的释放；而从图7中的WesternBlotting图可以看到，金属配合物二乙基二硫羰氨酸铜盐可以显著减少由脂多糖+尼日利亚菌素诱导引起的细胞焦亡执行分子GSDMD和Caspase-1的剪切，以及炎性因子IL-1β的剪切，而对NLRP3组分的表达没有明显的影响。
实验例2
金属配合物在防治脓毒症中的效果
将C57BL/6J小鼠26只(8周龄、雄性)称体重、编号后随机分为4组：生理盐水组、脂多糖模型组、脂多糖+金属配合物1mg/kg治疗组、脂多糖+金属配合物2mg/kg治疗组。适应性喂养3天后，脂多糖模型组、脂多糖+金属配合物1mg/kg治疗组、脂多糖+金属配合物2mg/kg治疗组予以腹腔注射30mg/kg的脂多糖溶液，生理盐水组同时予以腹腔注射同体积的生理盐水作为对照，而脂多糖+金属配合物1mg/kg治疗组、脂多糖+金属配合物2mg/kg治疗组在造模前4h予以腹腔注射相对应浓度的金属配合物溶液治疗。造模后第12h对小鼠予以1％的戊巴比妥钠麻醉后，打开小鼠胸腔，利用1mL注射器从小鼠左心室抽取小鼠血液，通过ELISA法检测小鼠血清中炎症因子IL-1β、IL-6水平。
实验结果如图8、图9所示，腹腔注射30mg/kg脂多糖12h后，相比于生理盐水组，脂多糖模型组和脂多糖+金属配合物组小鼠血清中的炎症因子IL-1β、IL-6水平升高；脂多糖+金属配合物组和脂多糖模型组相比，图8表明IL-6水平无明显统计学差异，由图9可知NLRP3特异性炎症因子IL-1β的水平显著下降，证明金属配合物二乙基二硫羰氨酸铜盐可以在小鼠脓毒症模型中显著抑制炎症因子IL-1β的释放。
实验例3
金属配合物对脂多糖诱导的脓毒症小鼠肺部病理损伤的影响
将实验例2抽取完血液的小鼠取左肺，固定于4％多聚甲醛，右肺留存于-80℃冰箱中，对取出的小鼠左肺进行脱水-浸蜡-包埋-切片-脱蜡-H&E染色-脱水封片处理，最后在光学显微镜下观察肺部病理损伤情况。
实验结果如图10所示，从小鼠的肺部H&E图中可观察到：生理盐水组小鼠肺部未见明显病理变化，脂多糖模型组小鼠肺部出现较明显的肺泡塌陷、肺泡壁增厚以及炎症细胞浸润，脂多糖+金属配合物组小鼠肺泡结构紊乱及炎症细胞浸润程度较模型组明显减轻，由图10可知，金属配合物二乙基二硫羰氨酸铜盐对于缓解脓毒症引发的肺部损伤具有明显的缓解作用。
实验结论
实施例1制备的金属配合物二乙基二硫羰氨酸铜盐该炎性小体抑制剂可直接抑制NLRP3炎性小体组装，达到抑制NLRP3炎性小体活化的效果，进而减少炎症因子IL-1β的释放，最终可以起到抑制细胞焦亡的效果，明显减轻NLRP3炎性小体活化引起炎症风暴和器官损伤，为制备治疗NLRP3炎性小体活化疾病药物提供了全新的选择。
该抑制剂可以抑制NLRP3炎性小体活化诱导的炎症因子释放及细胞焦亡，从而减少脓毒症小鼠肺泡塌陷、肺泡壁增厚以及炎症细胞浸润的病理损伤、降低小鼠血清NLRP3特异性炎症因子水平。该结果说明，该抑制剂二乙基二硫羰氨酸铜盐能够用于防治脓毒症等由炎性小体引起的相关疾病的治疗药物制备。
显然，上述实施例仅仅是为清楚地说明所作的举例，而并非对实施方式的限定。对于所属领域的普通技术人员来说，在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
Claims (8)
1.一种NLRP3炎性小体抑制剂，其特征在于，所述NLRP3炎性小体抑制剂为二乙基二硫羰氨酸铜盐。
2.根据权利要求1所述的NLRP3炎性小体抑制剂，其特征在于，所述二乙基二硫羰氨酸铜盐由二硫卡钠和铜离子反应形成。
3.根据权利要求1所述的NLRP3炎性小体抑制剂，其特征在于，所述NLRP3炎性小体抑制剂可抑制NLRP3炎性小体的活化。
4.根据权利要求1所述的NLRP3炎性小体抑制剂，其特征在于，所述NLRP3炎性小体抑制剂可抑制炎症因子IL-1β的释放。
5.根据权利要求1所述的NLRP3炎性小体抑制剂，其特征在于，所述NLRP3炎性小体抑制剂可抑制细胞焦亡。
6.一种如权利要求1～5所述的NLRP3炎性小体抑制剂在制备治疗NLRP3炎性小体活化疾病药物中的应用。
7.根据权利要求6所述的应用，其特征在于，所述NLRP3炎性小体活化疾病为脓毒症、炎症性肠炎、非酒精性脂肪肝、中风、痛风、糖尿病、动脉粥样硬化、阿尔兹海默症、哮喘中的一种。
8.根据权利要求6所述的应用，其特征在于，所述NLRP3炎性小体活化相关疾病药物治疗药物的剂型为注射剂、口服剂或外用剂。
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5886220A (en) * | 1994-09-09 | 1999-03-23 | Idemitsu Petrochemical Co., Ltd. | Process for preventing polymerization of vinyl compound |
| WO2000000192A1 (en) * | 1998-06-26 | 2000-01-06 | University Of Washington | INHIBITION OF NF-λB MEDIATED TISSUE INJURY USING DITHIOCARBAMATE DERIVATIVES |
| US20110206756A1 (en) * | 2008-03-17 | 2011-08-25 | University Of Utah Research Foundation | Dithiocarbamate metal chelates and methods of making and using thereof |
| US20120270850A1 (en) * | 2006-11-20 | 2012-10-25 | University Of Medlbourne | Metal delivery agents and therapeutic uses of the same |
| US20180000755A1 (en) * | 2015-01-06 | 2018-01-04 | The University Of Tokyo | Agent for controlling cells constituting cancer microenvironment or inflammatory microenvironment |
| US20190117595A1 (en) * | 2016-04-13 | 2019-04-25 | Nuoma (Beijing) Technology Co., Ltd | Method for treating pleuroperitoneal membrane cancers by locally injecting disulfiram preparation |
| CN110575453A (zh) * | 2019-10-30 | 2019-12-17 | 中国科学技术大学 | Ns1619在抑制nlrp3炎症小体活化中的应用 |
| US11033516B1 (en) * | 2020-09-18 | 2021-06-15 | Spring Discovery, Inc. | Combination therapies with disulfiram |
-
2022
- 2022-01-04 CN CN202210001768.5A patent/CN116421594A/zh active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5886220A (en) * | 1994-09-09 | 1999-03-23 | Idemitsu Petrochemical Co., Ltd. | Process for preventing polymerization of vinyl compound |
| WO2000000192A1 (en) * | 1998-06-26 | 2000-01-06 | University Of Washington | INHIBITION OF NF-λB MEDIATED TISSUE INJURY USING DITHIOCARBAMATE DERIVATIVES |
| US20120270850A1 (en) * | 2006-11-20 | 2012-10-25 | University Of Medlbourne | Metal delivery agents and therapeutic uses of the same |
| US20110206756A1 (en) * | 2008-03-17 | 2011-08-25 | University Of Utah Research Foundation | Dithiocarbamate metal chelates and methods of making and using thereof |
| US20180000755A1 (en) * | 2015-01-06 | 2018-01-04 | The University Of Tokyo | Agent for controlling cells constituting cancer microenvironment or inflammatory microenvironment |
| US20190117595A1 (en) * | 2016-04-13 | 2019-04-25 | Nuoma (Beijing) Technology Co., Ltd | Method for treating pleuroperitoneal membrane cancers by locally injecting disulfiram preparation |
| CN110575453A (zh) * | 2019-10-30 | 2019-12-17 | 中国科学技术大学 | Ns1619在抑制nlrp3炎症小体活化中的应用 |
| US11033516B1 (en) * | 2020-09-18 | 2021-06-15 | Spring Discovery, Inc. | Combination therapies with disulfiram |
Non-Patent Citations (1)
| Title |
|---|
| D. H. BROWN ET AL.: "Antiinflammatory Effects of Some Copper Complexes", JOURNAL OF MEDICINAL CHEMISTRY, vol. 23, no. 7, 31 July 1980 (1980-07-31), pages 729 - 734, XP001009751, DOI: 10.1021/jm00181a006 * |
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