CN116283533A - 具有nlrp3抑制活性的鸡脚参酮a及其衍生物与应用 - Google Patents
具有nlrp3抑制活性的鸡脚参酮a及其衍生物与应用 Download PDFInfo
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Abstract
本发明公开了具有NLRP3抑制活性的鸡脚参酮A及其衍生物与应用。鸡脚参酮A及其衍生物为松香烷二萜类化合物,对于NLRP3信号通路具有明显的抑制作用,可以有效地抑制J774A.1巨噬细胞LDH的释放,能显著抑制尼日利亚菌素诱导的NLRP3炎症小体活化后Caspase‑1的加工成熟、以及IL‑1β释放,能显著减少J774A.1巨噬细胞焦亡的比例。鸡脚参酮A及其衍生物可用作NLRP3抑制剂,可以用于NLRP3炎症小体介导的各类疾病的药物或产品的制备。
Description
技术领域
本发明属于药物化学领域,具体涉及具有NLRP3抑制活性的鸡脚参酮A及其衍生物与应用。
背景技术
炎症小体是因外源性微生物入侵或机体对内源性损伤信号的响应而形成,是由NOD样受体蛋白、ASC(凋亡相关点状蛋白)和Caspase-1(半胱氨酸天冬氨酸酶-1)组成的细胞质超分子复合物。目前报道的炎症小体类型共有十余种,其中NLRP3炎症小体(Nod样受体3)研究最深入,主要分布于巨噬细胞、树突状细胞中,与感染性疾病、体内损伤相关,且与免疫系统疾病关联性最大。
NLRP3炎症小体的活化需要双信号的参与:第一信号是通过TLRs或TNF受体活化转录因子NF-κB,从而上调NLRP3的表达,为第二信号发挥作用提供物质条件;第二信号是由ATP、尿酸盐和一些代谢产物(如葡萄糖、β淀粉样蛋白、氧化型低密度脂蛋白)、二氧化硅等刺激产生,能够促进NLRP3炎症小体组装,并激活Caspase-1p20(或p10)、GSDMDNT,进而促进IL-1β、IL-18、HMGB1、LDH等蛋白的成熟并释放到胞外,同时导致细胞焦亡。细胞焦亡所释放的细胞因子或蛋白会进一步激活自身免疫反应和获得性免疫反应。NLRP3炎症小体是固有免疫和获得性免疫的重要媒介,是炎症相关疾病的理想药物靶标。类风湿性关节炎、痛风、克罗恩病、冷吡啉综合征、II型糖尿病等多种疾病的发生与NLRP3炎症小体的异常活化密切相关。在小分子化合物中,MCC950作为NLRP3炎症小体特异性抑制剂的研究最为突出,曾作为治疗类风湿性关节炎药物进入II期临床实验,但因其产生肝脏毒性而未能通过临床实验。因此,研发具有低毒性、高活性的天然药物小分子具有重要的意义。
唇形科鸡脚参属植物鸡脚参(Orthosiphonwulfenioides)具有祛风除湿、清肺润燥、益阴敛汗、镇痛化积、接骨生肌的功能,用于风湿痛、消化不良、食积、虚弱头晕、虚汗、咳嗽;并用于脉管炎及骨折的治疗。罗禹等学者研究发现鸡脚参根醇提物的急性毒性较小,抗炎、止咳作用明显,具体表现为对大鼠蛋清性足肿胀、小鼠耳廓肿胀有明显的抑制作用,并且可延长二氧化硫引起的小白鼠咳嗽的潜伏期(罗禹,丁立生,田军,吴锷凤.鸡脚参醇提物药理活性研究.天然产物研究与开发.2003,15,216-218)。虽然鸡脚参的根具有显著的抗炎活性,民间用其治疗炎症性疾病,但未见有抗炎活性的化学成分报道,更没有发现具有抗炎作用的新化合物。
发明内容
本发明的目的在于提供具有NLRP3抑制活性的鸡脚参酮A及其衍生物与应用,具体为提供了具有松香烷型二萜骨架结构的新颖化合物(鸡脚参酮A)及其衍生物,其能有效地抑制J774A.1巨噬细胞的LDH释放,能显著抑制尼日利亚菌素诱导的NLRP3炎症小体活化后Caspase-1的加工成熟、以及IL-1β释放,并能显著减少J774A.1巨噬细胞焦亡的比例。所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物作为NLRP3抑制剂的药物组合物,用于治疗炎症小体介导的各类疾病。本发明为NLRP3炎症小体抑剂的开发提供了新的选择。
本发明是通过如下的技术方案得以实现的:
本发明提供具有NLRP3抑制活性的鸡脚参酮A及其衍生物,所述鸡脚参酮A及其衍生物具有松香烷二萜结构,结构式如下所示:
进一步地,所述鸡脚参酮A衍生物为以鸡脚参酮A为前体的化合物,包括鸡脚参酮A的酯化物、醚化物、卤代物、氰基化物、硝基化物、氨基衍生物及其盐类中的一种以上。
进一步地,所述鸡脚参酮A及其衍生物是通过提取分离或人工合成获得。
本发明还提供所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用。
进一步地,所述药物组合物中包含鸡脚参酮A及其衍生物(鸡脚参酮A的酯基、醚基、卤素、氰基、硝基、氨基衍生物及其盐类)中的一种以上。
进一步地,所述药物组合物为预防或治疗NLRP3炎症小体介导的各类疾病的药物组合物。
本发明的具有NLRP3抑制活性的鸡脚参酮A及其衍生物可以作为NLRP3炎症小体抑制剂用于治疗类风湿性关节炎、痛风、克罗恩病、冷吡啉综合征、II型糖尿病的新型药物。
进一步地,所述药物组合物的剂型包括固体制剂、半固体制剂、液体制剂、气体制剂。
进一步地,所述固体制剂包括片剂、胶囊剂、丸剂、颗粒剂,散剂,膜剂;所述半固体制剂包括软膏剂、眼膏剂、凝胶剂、栓剂、滴丸;所述液体制剂包括注射剂、洗剂、滴眼剂;所述气体制剂包括喷雾剂、气雾剂、粉雾剂。
进一步地,注射剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上加入注射用溶媒,精滤,灌封灭菌后可制成注射剂。
进一步地,喷雾剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上溶于溶媒中制成每毫升含有药物成分1-100mg的澄明溶液,分装于喷雾剂装置中。
进一步地,软膏剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上作为药物活性成分,使用油脂性和水溶性基质作为制备药物软膏剂的辅料,按一定比例制成每克含有药物成分1-100mg的软膏剂。
进一步地,凝胶剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上作为药物活性成分,加入能形成凝胶的材料作为制备凝胶剂的辅料,按一定比例制成每克含有药物成分1-100mg的凝胶剂。
进一步地,片剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上配以各种药用辅料可制成片剂。使用鸡脚参酮A及其衍生物中的一种以上作为药物活性成分,使用几种赋形剂作为制备组合药物片剂的辅料成分,按照一定比例配比制成每片含有药物成分1-100mg的片剂样品。
进一步地,胶囊剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上配以各种药用辅料可制成胶囊剂。使用鸡脚参酮A及其衍生物中的一种以上作为药物活性成分、使用几种赋形剂作为制备组合药物胶囊剂的辅料成分,按照一定比例配比制成每粒胶囊中含有化合物成分1-100mg的胶囊制剂。
进一步地,颗粒剂的制备方法为将鸡脚参酮A及其衍生物中的一种以上作为药物活性成分,使用赋形剂作为制备组合药物颗粒剂的辅料成分,按一定比例加入赋形剂制成每克含有化合物成分1-100mg的颗粒剂。
与现有技术相比,本发明具有如下优点和有益效果:
1、本发明中的鸡脚参酮A及其衍生物为高度氧化的松香烷二萜新颖化合物。
2、本发明中的鸡脚参酮A具有显著的LDH抑制活性,其IC50值为0.23±0.12μM。
3、本发明中的鸡脚参酮A通过抑制Caspase-1和IL-1β的表达选择性地阻断由于NLRP3过度激活所引起的细胞焦亡。
4、本发明中的鸡脚参酮A及其衍生物可广泛应用在NLRP3介导的各类疾病的防治药物或产品的制备。
附图说明
图1为鸡脚参酮A的结构式。
图2为鸡脚参酮A的高分辨质谱图。
图3为鸡脚参酮A的核磁氢谱图。
图4为鸡脚参酮A的核磁碳谱图。
图5为鸡脚参酮A的核磁COSY谱图。
图6为鸡脚参酮A的核磁HSQC谱图。
图7为鸡脚参酮A的核磁HMBC谱图。
图8为鸡脚参酮A的核磁ROESY谱图。
图9为鸡脚参酮A的紫外图谱(UV)。
图10为鸡脚参酮A的圆二色谱图(CD)。
图11为鸡脚参酮A的红外(IR)谱图。
图12为鸡脚参酮A的单晶衍射图。
图13为鸡脚参酮A对LDH释放率筛选柱形图。
图14为鸡脚参酮A对Caspase-1蛋白表达的抑制作用图。
图15为鸡脚参酮A对IL-1β蛋白表达的抑制作用图。
图16为鸡脚参酮A抑制单核巨噬细胞J774A.1焦亡图。
图17为衍生物脚参酮A1-A7对单核巨噬细胞J774A.1中LDH释放的影响图。
具体实施方式
下面结合实施例对本发明作进一步描述。
实施例1具有NLRP3抑制活性的鸡脚参酮A的制备及结构鉴定
鸡脚参(20kg)粉碎后用95%乙醇溶液(体积百分比浓度)在60℃回流提取3次,每次100L,每次12小时,回收溶剂后得浸膏(2kg)。浸膏(2kg)用30℃的温水稀释后用乙酸乙酯萃取3次,得乙酸乙酯萃取部位(0.6kg)。乙酸乙酯萃取部位(0.6kg)经大孔树脂(DM1O1)柱色谱(甲醇-水:体积比30%-100%)分离得到四个组份(Fr.A-Fr.D)。Fr.C(70g)经硅胶柱色谱(石油醚-乙酸乙酯,体积比10:0,10:1,5:1,1:1,1:5,1:10,0:10)分离得到七个组份(Fr.C.1-Fr.C.7)。Fr.C.2(7g)经MCIgelCHP20P柱色谱分离(甲醇-水,体积比75%-100%)得六个组份(Fr.C.2.1-Fr.C.2.6)。然后Fr.C.2.1(2.8g)用硅胶柱色谱(石油醚/丙酮(20:1,v/v)洗脱的)进一步分离,得到Fr.C.2.1.1-Fr.C.2.1.9九个组分。Fr.C.2.2.1(25mg)采用半制备高效液相色谱(甲醇与水的体积比为65:35,v/v)纯化得到鸡脚参酮A(2.5mg,tR=12.3min,3.0mL/min)。通过NMR,HRESIMS,CD,UV,IR等波谱数据,确定鸡脚参酮A的结构如图1。
鸡脚参酮A为黄色针状结晶,由高分辨质谱HRESIMS(图2)m/z333.2057[M+H]+(calcdforC20H29O4,333.2060)可知分子量为332,再结合核磁氢谱(图3)和碳谱(图4)推导出分式为C20H28O4。此外通过二维核磁COSY(图5),HSQC(图6),HMBC(图7),ROESY(图8)谱图确定了鸡脚参酮的平面结构及其相对构型。鸡脚参酮A的1H和13C NMR数据:1H NMR(400MHz,acetone-d6):H2-1(δH2.09m,1.66m),H2-2(δH1.68m,1.43m),H2-3(δH1.39m,1.27m),H-5(δH1.87d,J=10.2Hz),H-6(δH4.42dd,J=10.2,2.8Hz),H-7(δH6.37d,J=2.8Hz),H-14(δH7.12s),H-15(δH2.86sept,J=6.9Hz),H3-16(δH1.06d,J=6.9Hz),H3-17(δH1.11d,J=6.9Hz),H3-18(δH1.19s),H3-19(δH1.12s),H3-20(δH1.15s);13C NMR(100MHz,acetone-d6):δC193.6(C-11),188.2(C-12),144.4(C-13),144.2(C-7),143.3(C-14),135.1(C-8),78.9(C-9),68.7(C-6),48.2(C-5),44.3(C-3),42.4(C-10),36.9(C-18),34.0(C-4),32.8(C-1),27.6(C-15),23.1(C-19),21.8(C-17),21.6(C-16),19.2(C-2),17.7(C-20)。
鸡脚参酮A的其它波普数据如下:UV(MeOH)λmax(logε):190(4.27),248(3.62),and304(3.42)nm(图9).CD(MeOH)λmax(Δε)208(-2.7),242(+1.5),312(-3.21),and344(+2.89)nm(图10)。IR(KBr)νmax;3533,3435,2981,2957,2923,1730,1671,1639,1456,1385,1230,1160,1113,1019cm-1(图11)。
最后,通过单晶X-ray衍射分析确定了鸡脚参酮A的绝对构型,如图12所示。
实施例2鸡脚参酮A的乳酸脱氢酶(LDH)抑制活性。
利用J774A.1巨噬细胞由脂多糖(LPS)和尼日利亚菌素(Nigericin)共诱导构建NLRP3炎症小体活化模型。将J774A.1巨噬细胞放于37℃、5mg/LCO2培养箱中培养,以5×105个细胞铺于6孔板中,培养细胞16h。观察细胞状态良好,吸走上清液,加入用Opti-MEM配制的200ng/mL的脂多糖(LPS)刺激3h,使第一信号产生。移去上清液,再加入Opti-MEM配制的10μM鸡脚参酮A处理30min,MCC950作为阳性药。随后,加入用Opti-MEM配制的10μM尼日利亚菌素(Nigericin)刺激1h,使第二信号产生。观察细胞,J774A.1巨噬细胞NLRP3炎症小体激活。
J774A.1巨噬细胞NLRP3炎症小体激活后,收集细胞培养液上清液于酶标板中。再加入LDH检测工作液,混匀,孵育15min。在490nm处测定吸光度,630nm处作为参考波长进行双波长测定。按照公式进行计算,即LDH释放率=(处理样品吸光度-经DMSO处理的对照细胞孔吸光度)/(细胞最大酶活性的吸光度-经DMSO处理的对照细胞孔吸光度)×100%。
所有实验至少重复3次。实验数据采用Excel和Graph Pad Prism7.0(Graph PadSoftware,San Diego,CA,USA)进行分析计算。采用单因素方差分析(One-wayANOVA)对各组间的差异进行分析,结果统计用数据平均值加减标准差means±S.D.表示。当P值<0.05为有统计学意义,具有显著性差异。
从图13可知鸡脚参酮A能抑制J774A.1巨噬细胞的LDH释放,且释放率小于30%。
复筛结果显示:鸡脚参酮A对LDH抑制率的IC50值为0.23±0.12μM,阳性对照MCC950的IC50值为0.03±0.02μM。
实施例3鸡脚参酮A显著抑制NLRP3炎症小体活化后Caspase-1的加工成熟及IL-1β释放。
上清蛋白提取:J774A.1巨噬细胞的NLRP3炎症小体激活后,收集细胞培养上清液于EP管中,加入去离子水配制的20g/100mL三氯醋酸(TCA)溶液(培养液上清与三氯醋酸溶液的体积比为1:1)提取上清蛋白,冰上静置30min。在12000g,4℃条件下离心15min,弃去上清液。加入与弃去上清液等体积冰浴的丙酮(0℃),在12000g,4℃条件下离心5min,弃去上清液。重复加入冰丙酮并离心倒走上清液的步骤两次,即得上清蛋白。按10倍于上清液比例加入2×Laemli Buffer,在沸水浴条件下加热10min。
细胞裂解液蛋白提取:吸走细胞培养液上清,向细胞中加入300μL含苯甲基磺酰氟(PMSF)的RIPA裂解液(RIPA:PMSF=1000:1)充分裂解15s,转移至EP管中。冰上裂解30min,12000g,4℃离心5分钟,收集上清液于EP管中。用超声细胞破碎仪充分破碎细胞,按照5倍于上清液的比例加入6×SDS-PAGE Loading Buffer,在沸水浴100℃条件下加热10min。
将上清蛋白提取样品和细胞裂解液蛋白提取样品分别进行聚丙烯酰胺凝胶(SDS-PAGE)电泳,使用湿转移系统将蛋白转移到硝酸纤维素(NC)膜上。随后,用50g/L的脱脂奶粉封闭1h,再用1×TBST洗膜三次,每次5min。上清蛋白在相应条带处孵育Caspase-1和IL-1β一抗,细胞裂解液蛋白在相应条带处孵育β-actin一抗,配合滚轴混合器12h。第二天,用1×TBST洗膜三次,在50g/L脱脂奶粉中加入相应二抗在摇床摇动1h,用1×TBST洗膜四次。最后,在膜上相应条带处均匀涂抹ECL化学发光剂,在凝胶成像仪中曝光。
所有实验至少重复3次。实验数据采用Excel和Graph Pad Prism 7.0(Graph PadSoftware,San Diego,CA,USA)进行分析计算。采用单因素方差分析(One-way ANOVA)对各组间的差异进行分析。当P值<0.05为有统计学意义,具有显著性差异。
从图14和图15可知,鸡脚参酮A在0.1-1μmol能剂量依赖性的减少Caspase-1成熟和IL-1β的释放。
实施例4鸡脚参酮A显著减少J774A.1巨噬细胞的焦亡。
J774A.1巨噬细胞的NLRP3炎症小体激活后,移去上清液,加入PBS配制的PI(3μg/mL)和Hochest3342(0.5μg/mL)染料,在37℃培养箱中孵育5min,用荧光倒置显微镜进行拍照。所有实验至少重复3次。实验数据采用Excel和GraphPadPrism7.0(GraphPadSoftware,SanDiego,CA,USA)进行分析计算。采用单因素方差分析(One-wayANOVA)对各组间的差异进行分析。当P值<0.05为有统计学意义,具有显著性差异。
从图16可知,分别用PI和Hoechst3342试剂对巨噬细胞J774A.1进行染色,PI使焦亡细胞染成红色,Hoechst3342使细胞核染呈蓝色。鸡脚参酮A在0.1-1μmol剂量下能减少PI染色的比例,因此说明该化合物能有效的抑制细胞焦亡引起的细胞膜打孔,从而减少细胞焦亡。
实施例5鸡脚参酮A衍生物的制备
1.酯化衍生物的制备
鸡脚参酮A(10mg)溶于3mLCH2Cl2溶剂中,于室温下加入乙酸酐(20μL)、三乙胺(30μL)、DMAP(4-二甲氨基吡啶,3.0mg)并室温(25℃)反应2h。向反应液中加入饱和碳酸氢钠水溶液5mL,用乙酸乙酯萃取,有机相用饱和NaCl水溶液洗涤,无水硫酸钠干燥,减压浓缩得黄色油状物。经硅胶柱层析,石油醚∶乙酸乙酯=10:1(体积比),得到鸡脚参酮A酯化衍生物,即鸡脚参酮A1(8.3mg)。
2.醚化衍生物的制备
鸡脚参酮A(10mg)溶于3mL甲醇溶剂中,加入对甲苯磺酸(3mg),于室温反应8h。向反应液中加水15mL,用乙酸乙酯萃取,有机相用饱和NaCl水溶液洗涤,无水硫酸钠干燥,减压浓缩得淡黄色粉末。经硅胶柱层析(石油醚:丙酮=体积比为10:1)得到鸡脚参酮A2(7.4mg)。
3.卤化衍生物的制备
鸡脚参酮A(10mg)溶解于乙酰氯(AcCl)(0.5mL)中,在0℃冰浴滴入甲醇(0.5mL)搅拌17小时,然后蒸干溶剂,得到鸡脚参酮A3(8.1mg)粉末。
4.氰化衍生物的制备
鸡脚参酮A3(10mg)溶解于二甲基亚砜(1.5mL)中,再加入NaCN(0.5mg),在冰水浴半固态搅拌1.5h过滤沉淀,获得鸡脚参酮A4(6.9mg)。
5.环氧化衍生物的制备
将鸡脚参酮A(10mg)溶解在二氯甲烷(DCM,2mL)中,再加入间氯过氧苯甲酸(m-CPBA,1.3mg),回流加热48小时,然后冷却至室温。用DCM(10mL)稀释滤液,然后用饱和Na2SO3水溶液(10mL)、饱和NaHCO3水溶液(10mL)和NaCl水溶液(10mL)洗涤。有机层在MgSO4上干燥。经硅胶柱层析,石油醚:乙酸乙酯=9:1(积比),得到鸡脚参酮A5(6.0mg)。
6.氨基化衍生物的制备
在微波反应管中加入环氧化物鸡脚参酮A5(10mg)和氢氧化铵水溶液(NH4OH水溶液,体积分数30%,0.4mL)。将微波反应管密封,85℃微波辐照30分钟,反应冷却至室温,旋转蒸发去除溶剂。在真空下干燥得到氨基醇鸡脚参酮A6(8.5mg)。
7.酰胺衍生物的制备
鸡脚参酮A6(10mg)溶于2mL乙醚(Et2O)溶剂中,于室温下加入水(1mL)和氢氧化钾(3mg),并(0℃)加入乙酰氯(0.1mL)反应15min。向反应液中加入饱和碳酸氢钠水溶液5mL,用乙酸乙酯萃取,有机相用饱和NaCl水溶液洗涤,无水硫酸钠干燥,减压浓缩得黄色粉末。重结晶得到鸡脚参酮A酰胺衍生物,即鸡脚参酮A7(6.8mg)。
实施例6鸡脚参酮A1-A7对单核巨噬细胞J774A.1中乳酸脱氢酶(LDH)的抑制活性。
将J774A.1巨噬细胞放于37℃、5mg/LCO2培养箱中培养,以5×105个细胞铺于6孔板中,培养细胞16h。观察细胞状态良好,吸走上清液,加入用Opti-MEM配制的200ng/mL的脂多糖(LPS)刺激3h,使第一信号产生。移去上清液,再加入Opti-MEM配制的10μM鸡脚参酮A1-A7处理30min,MCC950作为阳性药。随后,加入用Opti-MEM配制的10μMNigericin刺激1h,使第二信号产生。观察细胞,J774A.1巨噬细胞NLRP3炎症小体激活。
J774A.1巨噬细胞NLRP3炎症小体激活后,收集细胞培养液上清液于酶标板中。再加入LDH检测工作液,混匀,孵育15min。在490nm处测定吸光度,630nm处作为参考波长进行双波长测定。按照公式进行计算,即LDH释放率=(处理样品吸光度-经DMSO处理的对照细胞孔吸光度)/(细胞最大酶活性的吸光度-经DMSO处理的对照细胞孔吸光度)×100%。
所有实验至少重复3次。实验数据采用Excel和GraphPadPrism7.0(GraphPadSoftware,SanDiego,CA,USA)进行分析计算。采用单因素方差分析(One-wayANOVA)对各组间的差异进行分析,结果统计用数据平均值加减标准差means±S.D.表示。当P值<0.05为有统计学意义,具有显著性差异。
从图17可知鸡脚参酮A1-A7能抑制J774A.1巨噬细胞的LDH释放,且释放率小于30%,复筛结果如下表所示:
实施例7:注射剂的制备
用实施例1中鸡脚参酮A加入注射用溶媒,精滤,灌封灭菌后制成每毫升含有药物成分1-100mg的注射液。
实施例8:喷雾剂的制备
将实施例1中鸡脚参酮A溶于乙醇中制成每毫升含有药物成分1-100mg的澄明溶液,分装于喷雾剂装置中。
实施例9:软膏剂的制备
将实施例1中鸡脚参酮A作为药物活性成分,使用油脂性和水溶性基质作为制备药物软膏剂的辅料,按鸡脚参酮A与基质质量比为(1:5)-(1:10)的配比制成每克含有药物成分1-100mg的软膏剂。
实施例10:凝胶剂的制备
将实施例1中鸡脚参酮A作为药物活性成分,加入能形成凝胶的材料作为制备凝胶剂的辅料,按鸡脚参酮A与凝胶质量比为(1:5)-(1:10)的配比制成每克含有药物成分1-100mg的凝胶剂。
实施例11:胶囊剂的制备
使用实施例1中鸡脚参酮A和实施例5中鸡脚参酮A1(质量比1:1)作为药物活性成分,使用赋形剂作为制备组合药物胶囊剂的辅料成分,按药物活性成分与赋形剂质量比为(1:5)–(1:10)的比例加入赋形剂制成每粒胶囊中含有化合物成分1-100mg的胶囊制剂。
实施例12:片剂的制备
使用实施例5中鸡脚参酮A1-A7(分别按照质量比1:1:1:1:1:1:1)作为药物活性成分,使用赋形剂作为制备药物片剂的辅料成分,按药物活性成分(鸡脚参酮A1-A7)与赋形剂质量比为(1:5)-(1:10)的配比制成每片含有药物成分1-100mg的片剂样品,再进行压片。
实施例13:颗粒剂的制备
使用实施例1中鸡脚参酮A和实施例5中鸡脚参酮A1-A7(分别按照质量比1:1:1:1:1:1:1:1)作为药物活性成分,使用赋形剂作为制备组合药物颗粒剂的辅料成分,按药物活性成分(鸡脚参酮A1-A7)与赋形剂质量比为(1:5)–(1:10)的比例加入赋形剂制成每克含有化合物成分1-100mg的颗粒剂。
Claims (10)
2.根据权利要求1所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物,其特征在于,鸡脚参酮A衍生物为以鸡脚参酮A为前体的化合物,包括鸡脚参酮A的酯化物、醚化物、卤代物、氰基化物、硝基化物、氨基衍生物及其盐类中的一种以上。
3.根据权利要求1所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物,其特征在于,所述鸡脚参酮A及其衍生物是通过提取分离或人工合成获得。
4.权利要求1-3任一项所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用。
5.根据权利要求4所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,所述药物组合物包括鸡脚参酮A及其衍生物中的一种以上。
6.根据权利要求4所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,其特征在于,所述药物组合物为预防或治疗NLRP3炎症小体介导的疾病的药物组合物。
7.根据权利要求4所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,其特征在于,所述药物组合物为治疗类风湿性关节炎、痛风、克罗恩病、冷吡啉综合征、II型糖尿病的药物组合物。
8.根据权利要求4所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,其特征在于,所述药物组合物的剂型包括固体制剂、半固体制剂、液体制剂、气体制剂。
9.根据权利要求8所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,其特征在于,所述所述固体制剂包括片剂、胶囊剂、丸剂、颗粒剂,散剂,膜剂;所述半固体制剂包括软膏剂、眼膏剂、凝胶剂、栓剂、滴丸。
10.根据权利要求8所述具有NLRP3抑制活性的鸡脚参酮A及其衍生物在药物组合物中的应用,其特征在于,所述液体制剂包括注射剂、洗剂、滴眼剂;所述气体制剂包括喷雾剂、气雾剂、粉雾剂。
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