CN117417331A - 一种基质调控类药物-半花菁的偶联物及其制备方法与应用 - Google Patents
一种基质调控类药物-半花菁的偶联物及其制备方法与应用 Download PDFInfo
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Abstract
本发明公开了一种基质调控类药物‑半花菁的偶联物。所述偶联物的化学结构如式(Ⅰ)所示;其中,R1为基质调控类药物,R2为刺激响应的连接分子,R3为C1~C18的烷基或烷基磺酸。本发明还公开了该偶联物的制备方法以及在抗肿瘤药物中的应用。本发明提供的一种基质调控类药物‑半花菁的偶联物,具有可在肿瘤部位刺激响应性释放的特性,从而释放出基质调控类药物以及半花菁;一方面,基质调控类药物可直接实现对肿瘤相关成纤维细胞的功能调控,另一方面,半花菁可在激光照射下呈现温和光热的特性从而实现对肿瘤相关成纤维细胞的功能调控,从而解决肿瘤部位致密的细胞外基质造成临床药物在肿瘤部位富集少与难以深部穿透的难题。
Description
技术领域
本发明属于生物医用材料领域和医药技术领域,更具体地,涉及一种基质调控类药物-半花菁的偶联物及其制备方法与应用。
背景技术
实体瘤细胞致密的细胞外基质(extracellular matrix,ECM)会随着肿瘤的发生发展持续堆积,造成异常的肿瘤力学微环境,从而严重阻碍药物向肿瘤部位的递送及在肿瘤组织中深部传统。药物在肿瘤组织的递送和分布不足,不但影响抗肿瘤治疗效果,也是肿瘤耐药、复发和转移的重要原因。因此,通过调控肿瘤力学性能来促进药物递送,提高常规抗肿瘤治疗的效果越发成为肿瘤研究领域的研究重点。由此可见,肿瘤基质成分是形成肿瘤力学微环境的关键因素,而下调肿瘤基质的合成或分解肿瘤内现存的基质成分,是调控肿瘤力学性能的有效策略。
近年来,纳米药物介导的温和光热可以通过减少肿瘤相关成纤维细胞,从而改善肿瘤的细胞外基质,进一步降低肿瘤的固体应力,增强化疗药物的深部穿透和抗肿瘤疗效。同时不少小分子药物,如氯沙坦、曲尼斯特和吡非尼酮等可以通过下调肿瘤基质组分的合成或通过蛋白水解酶等直接分解肿瘤组织内的基质蛋白来调控肿瘤力学性能。
但目前还缺少一种能够实现温和光热联合基质调控药物的双重策略的药物,如果一旦实现,这对于改善肿瘤异常的力学微环境从而提高临床药物的抗肿瘤疗效具有重大的研究意义。
发明内容
针对现有技术的以上缺陷或改进需求,本发明提供了一种多功能基质调控类药物-半花菁偶联物的制备和应用,该偶联物单分子能自组装为纳米粒,系统给药后高浓度富集于肿瘤部位,通过肿瘤部位的内源性刺激响应性地释放出基质调控药物以及半花菁染料,其中基质调控类药物可直接改善肿瘤力学,而半花菁染料在激光照射下能够产生温和光热来改善肿瘤力学,依靠二者的联合作用来双重解决临床药物在肿瘤部位由于致密的细胞外基导致临床药物在肿瘤部位蓄积不够的问题。
为实现上述目的,按照本发明的一个方面,提供了一种基质调控类药物-半花菁的偶联物,所述偶联物的化学结构如式(Ⅰ)所示:
其中,R1为基质调控类药物,R2为刺激响应的连接分子,R3为C1~C18的烷基或烷基磺酸。
优选地,所述基质调控类药物为氯沙坦、曲尼斯特、吡非尼酮或卡泊三醇。
优选地,所述刺激响应的连接分子为ROS响应键、GSH响应键、酶响应键或pH响应键。
优选地,所述偶联物在分散液中以自组装形成的纳米颗粒的形式存在。
作为进一步优选地,所述分散液中偶联物的浓度为2~20mg/mL。
按照本发明的另一方面,提供了一种制备上述偶联物的方法,所述偶联物的合成路线如下:
作为进一步优选地,式(IV)化合物的合成方式如下:
作为更进一步优选地,式(Ⅲ)化合物的合成方式如下:
优选地,还包括利用纳米沉淀法获得式(Ⅰ)化合物的纳米颗粒分散液。
按照本发明的另一方面,还提供了上述的偶联物在抗肿瘤药物中的应用。
总体而言,通过本发明所构思的以上技术方案与现有技术相比,由于通过对合成路线的改进,获得了具有基质调控类药物和半花菁结构的偶联物,从而具有以下有益效果:
1.本发明提供的一种基质调控类药物-半花菁的偶联物,具有可在肿瘤部位刺激响应性释放的特性,从而释放出基质调控类药物以及半花菁;一方面,基质调控类药物可直接实现对肿瘤相关成纤维细胞的功能调控,另一方面,半花菁可在激光照射下呈现温和光热的特性从而实现对肿瘤相关成纤维细胞的功能调控;经验证该偶联物可实现针对肿瘤力学微环境的双重改善,从而解决肿瘤部位致密的细胞外基质造成临床药物在肿瘤部位富集少与难以深部穿透的难题;
2.本发明的基质调控类药物-半花菁的偶联物能够促进抗肿瘤药物在肿瘤部位的蓄积,从而提高抗肿瘤效果,其在调控肿瘤力学从而改善纳米药物在肿瘤部位递送和分布不足的难题方面有着巨大的临床潜力;
3.该基质调控类药物-半花菁偶联物,优选通过纳米沉淀法获得单分子自组装纳米药物递送系统,制备的纳米制剂稳定性良好,无需其他多余稳定剂进行稳定,制备工艺简单。
4.该偶联物在水中具有良好的稳定性、对肿瘤相关成纤维细胞具有更强的杀伤和功能调控以及肿瘤微环境刺激响应特性。
附图说明
图1为本发明实施例1的核磁共振氢谱;
图2为本发明实施例1的高分辨质谱;
图3为本发明实施例1制得的纳米颗粒在体外条件下ROS响应能力的检测的紫外吸收光谱对比(A)以及在体外的活性氧响应荧光恢复变化(B);
图4为本发明实施例1的透射电镜图(A)和粒子粒径图(B);
图5为本发明实施例1制得的纳米颗粒在不同环境中的Zeta电位测定结果;
图6a为本发明实施例1制得的纳米颗粒在超纯水中的粒径和聚合物分散性指数;
图6b为本发明实施例1制得的纳米颗粒在生理盐水中的粒径和聚合物分散性指数;
图6c本发明实施例1制得的纳米颗粒在磷酸缓冲盐溶液中的粒径和聚合物分散性指数;
图7为本发明实施例1制得的纳米颗粒的升温情况(A)以及在不同升温条件下的红外热成像仪的结果(B);
图8为本发明实施例1的纳米颗粒在24小时和48小时后,对肿瘤相关成纤维细胞CAFs、肿瘤相关成纤维细胞和正常细胞NIH 3T3的毒性影响;
图9为本发明实施例1在光照条件下的杀伤选择性效果;
图10为本发明实施例1与其它药物分别作用于患瘤小鼠的小鼠瘤体积-时间曲线(A)以及剥离后肿瘤照片(B);
图11为本发明实施例1的生物安全性研究结果,其中,(A)表示白细胞量,(B)表示红细胞量,(C)表示血小板量,(D)表示血红蛋白量;
图12为小鼠经本发明实施例1治疗后,小鼠的主要器官(心、肝、脾、肺、肾)的组织切片。
具体实施方式
为了使本发明的目的、技术方案及优点更加清楚明白,以下结合附图及实施例,对本发明进行进一步详细说明。应当理解,此处所描述的具体实施例仅仅用以解释本发明,并不用于限定本发明。此外,下面所描述的本发明各个实施方式中所涉及到的技术特征只要彼此之间未构成冲突就可以相互组合。
本发明提供了基质调控类药物-半花菁的偶联物,化学结构如式(Ⅰ)所示:
其中,R1为一类调控基质的药物如氯沙坦、曲尼斯特、吡非尼酮、卡泊三醇等及其(不影响其药物活性的)衍生物;
R2为一类刺激响应的连接分子,如ROS响应键、GSH响应键、酶响应键或pH响应键等;其中酶响应键里的酶指的是胞质酶、线粒体酶、溶酶体、核酸酶或蛋白酶。
R3选自C1~C18的烷基或烷基磺酸,优选为C1~C10的烷基或烷基磺酸,更佳地为C3~C6的烷基或烷基磺酸。
本发明还提供了一种上述基质调控类药物-半花菁的偶联物的制备方法,包括以下步骤:
步骤1:根据如下步骤,获得式(Ⅲ)化合物;
步骤2:根据如下步骤,获得式(IV)化合物;
步骤3:根据如下步骤,获得式(Ⅰ)化合物;
步骤4:利用纳米沉淀法,获得式(Ⅰ)化合物的纳米颗粒分散液;其具体为,将式(Ⅰ)化合物溶解在中性有机溶剂中,随后滴加至去离子水中进行搅拌,最后采用去离子水进行透析,得到自组装纳米粒分散液,即为所述基质调控类药物-半花菁的偶联物纳米制剂。
一些实施例中,所述中性性有机溶剂为二甲亚砜、乙醇、甲醇和四氢呋喃中的一种或多种。所述搅拌状态下的去离子水,其搅拌速度不低于400rpm,较佳为400~2000rpm。所述偶联物溶液中偶联物的浓度为2~20mg/mL,更佳为5~10mg/mL。所述透析袋截留分子量根据制备的目标偶联物的分子量确定,通常为1kDa~10kDa,共透析6~12h,中途换水两次。
该式(Ⅰ)化合物构成的的基质调控类药物-半花菁的偶联物纳米制剂能够增敏临床药物抗肿瘤疗效的作用,上述增敏临床药物包括但不限于为盐酸阿霉素脂质体注射液Doxil、奥沙利铂(Oxaliplatin)、紫杉醇(Paclitaxel)、多西他赛(Docetaxel)、吉西他滨(Gemcitabine)、卡培他滨(Capecitabine)、羟基喜树碱(Hydroxycam pothecin)、吡柔比星(Pirarubicin)、表柔比星(Epirubicin)等;上述肿瘤包括但不限于为乳腺癌、卵巢癌、胰腺癌、肝癌、结肠癌或黑色素瘤等。
本发明提供的一种基质调控类药物-半花菁偶联物,其作用主要是在水中具有良好的稳定性、对肿瘤相关成纤维细胞具有更强的杀伤和功能调控以及肿瘤微环境刺激响应特性。
以下为实施例:
实施例1
在本实施例中,R1为吡非尼酮(PFD),R2为具有ROS刺激响应的连接分子TK,R3为C4H8O3S-,
如图1所示,合成步骤包括:
S1.羟基化的吡非尼酮(PFD-OH)的制备
PFD-OH,即羟基修饰的基质调控类药物PFD的制备路线如上,其具体为:
S11.秤取吡非尼酮(185.5mg,1.0mmol)和N-溴丁二酰亚胺(NBS)(178.6mg,1.0mmol)混合在7.0mL四氯化碳(CCl4)中,然后添加偶氮二异丁腈(20.0mg,0.12mmol)。
S12.将上述反应液在90℃条件回流同时磁力搅拌2小时,并通过薄层层析法监测反应。将反应体系冰浴30分钟,促进固体析出。
S13.通过抽滤收集上述反应液体,并通过旋蒸干燥得到黄色粘稠液体产物溴代化的吡非尼酮(PFD-Br)。
S14.加入当量浓度4N的氢氧化钠10ml,40℃条件下搅拌4小时,通过薄层层析法监测反应,抽滤得到黄色液体,后处理分别用二氯甲烷、二氯甲烷+水萃取,饱和氯化钠洗涤,然后加入无水硫酸钠干燥,最后通过旋蒸浓缩后真空干燥,真空干燥的温度控制在20~50℃,得到PFD-OH。
S2.磺酸根取代的半花菁荧光药物(SO3 Cy)的制备
SO3 Cy的制备路线如上,其具体为:
将间苯二酚(88mg,0.8mmol)和K2CO3(110mg,0.8mmol)溶于1.5mL无水N,N-二甲基甲酰胺(DMF),室温下N2保护搅拌10min,然后将溶于2mL DMF中的2-[2-[2-氯-3-[2-[1,3-二氢-3,3-二甲基-1-(4-磺丁基)-2H-吲哚-2-亚基]亚乙基]-1-环己烯-1-基]乙烯基]-3,3-二甲基-1-(4-磺丁基)-3H-吲哚内盐钠盐IR783(300mg,0.4mmoL)加入上述反应体系,N2保护下,45℃条件下反应6h。利用油泵90℃条件旋蒸去除DMF,干燥后处理采用CH2Cl2萃取,饱和食盐水洗涤,无水硫酸钠干燥,浓缩,硅胶拌样,以CH2Cl2:MeOH=50:1、CH2Cl2:MeOH=20:1、CH2Cl2:MeOH=5:1依次为洗脱剂柱层析分离纯化样品,得到SO3 Cy。
S3.硫缩酮键TK-OH(即双羟基修饰的刺激响应的连接分子)的制备
TK-OH的制备路线如上,其具体为:
S31.将巯基乙酸(9.2g,100mmol)、丙酮(3.54g,60mmol)和10μL三氟乙酸(TFA)在室温下混合搅拌5h。然后将混合物放入冰浴中促进完全结晶。然后,将溶液离心后,用正己烷和水洗涤3次,得到产率为82%的白色产物。
S32.9000rpm,5min离心,将产物依次用石油醚、水、水、正己烷洗涤离心后,真空干燥得到白色产物TK-COOH。
S33.将化合物TK-COOH(1.5g,6.69mmol)溶解在无水四氢呋喃(THF)150mL中,冰浴搅拌,缓慢加入氢化铝锂(1.52g,40.1mmol),搅拌30min后,用N2吹扫,以防止积累氢气。
S34.去除冰浴后在室温条件下搅拌4h,然后在冰浴条件下依次缓慢滴加入冰水(1mL)、10%氢氧化钾水溶液、冰水,使反应中的氢化铝锂发生猝灭,搅拌5分钟后撤去冰浴,加无水硫酸镁(4.5g,37.4mmol)室温搅拌20分钟后抽滤,将滤液在45℃条件下旋蒸浓缩。
S35.用硅胶拌样,以石油醚:乙酸乙酯=1:1为洗脱剂柱层析分离纯化样品,得到TK-OH。
S4.将TK-OH与二(对硝基苯)碳酸酯(NPC)反应生成NPC-TK-OH
NPC-TK-OH的制备路线如上,其具体为:
TK-OH(300mg,1.5mmol)溶于5mL无水CH2Cl2中,加入300μL N,N-二异丙基乙胺(DIPEA),冰浴条件下加入溶于4mL无水CH2Cl2中的二(对硝基苯)碳酸酯(NPC),白烟消失后撤走冰浴,N2保护条件下室温搅拌4h,后处理采用CH2Cl2萃取,有机相用饱和盐水洗涤,无水硫酸钠干燥,浓缩,以石油醚:乙酸乙酯=3:1为洗脱剂柱层析分离纯化样品,得到NPC-TK-OH。
S5.将NPC-TK-OH与PFD-OH反应制备PFD-TK-OH
PFD-TK-OH的制备路线如上,其具体为:
步骤S1获得的PFD-OH(434mg,2.16mmol)溶于10mL无水CH2Cl2中,加4-二甲氨基吡啶DMAP(330mg,2.7mmol),室温搅拌条件下,加入溶于10mL无水CH2Cl2中的步骤S4获得的NPC-TK-OH(1g,2.7mmol),N2保护条件下室温搅拌过夜,后处理采用CH2Cl2萃取,有机相用饱和盐水洗涤,无水硫酸钠干燥,浓缩,以石油醚:乙酸乙酯=1:5为洗脱剂柱层析分离纯化样品,得到PFD-TK-OH。
S6.吡非尼酮硫缩酮半花菁(PFD-TK-Cy)的制备
S61.三光气溶于4mL无水CH2Cl2(145mg,0..5mmol)中,N2保护条件下冰浴搅拌,加入溶于10mL无水CH2Cl2中的PFD-TK-OH(100mg,0.24mmol)后加入50μL DIPEA,撤走冰浴,室温搅拌10分钟。通过薄层层析法监测反应结束后,旋蒸去除溶剂和未反应完的三光气。
S62.加入5mL无水CH2Cl2到上述体系中复溶,缓慢滴加溶于3mL无水CH2Cl2中的SO3Cy(50mg,0.1mmol),再加入20μLDIPEA。N2保护条件下室温搅拌5小时,通过薄层层析法监测反应。
S63.后处理采用CH2Cl2萃取,有机相用饱和盐水洗涤,无水硫酸钠干燥,浓缩,以CH2Cl2:MeOH=20:1、CH2Cl2:MeOH=10:1依次为洗脱剂柱层析分离纯化样品,得到最终产物PFD-TK-Cy,产物结构如下图所示。
图1和图2为实施例1制备得到的化合物的核磁共振氢谱和高分辨质谱。
S7.PFD-TK-Cy纳米颗粒的制备
PFD-TK-Cy纳米颗粒的自组装是自发进行的,采用纳米沉淀法制备。具体来说,取2mg PFD-TK-Cy溶于200μl DMSO,使得浓度为10mg/mL。滴加到2ml超纯水,使得DMSO:水(体积比)=1:10,吹打均匀。将上述混合体系转入到3.5K Da截留分子量的透析袋,透析6h,中途换水2次除去有机溶剂,得到的纳米粒子的分散液为蓝色澄清溶液。
验证例1 PFD-TK-Cy纳米颗粒的表征
图3展示了实施例1制得的PFD-TK-Cy纳米颗粒在体外条件下ROS响应能力的检测;双氧水作为ROS的一种,由于SO3 Cy骨架上酚羟基连接TK健后被修饰成酚酯键,分子内电荷转移(Intramolecular charge transfer,ICT)效应变弱,导致PFD-TK-Cy荧光强度远低于SO3 Cy,但是由于ROS介入后会导致PFD-TK-Cy的TK健断裂后释放出SO3 Cy,ICT效应增强,因此荧光也会逐渐恢复。其中,A为紫外吸收光谱对比,B为在体外的活性氧响应荧光恢复变化(B);结果显示,PFD-TK-Cy与双氧水共孵育后,随时间增加荧光也逐渐增加,说明ROS导致PFD-TK-Cy中TK键的断裂从而释放SO3 Cy,证明PFD-TK-Cy具有ROS响应的特性。
图4和图5展示了实施例1制得的PFD-TK-Cy纳米颗粒的透射电镜图(图4A)、粒子粒径图(图4B)以及纳米颗粒分别在超纯水、生理盐水、磷酸盐缓冲溶液中的Zeta电位测定结果(图5),结果表明,制得的PFD-TK-Cy纳米颗粒尺寸均一,平均直径为150nm左右,PFD-TK-Cy纳米颗粒表面电荷呈现弱中性。
图6展示了实施例1制得的PFD-TK-Cy纳米颗粒分别在超纯水(图6a)、生理盐水(图6b)、磷酸盐缓冲溶液(图6c)中的粒径和聚合物分散性指数的变化,结果表明,制得的PFD-TK-Cy纳米颗粒7天内粒径都维持在150nm左右,PDI也小于0.2,纳米粒具有良好的稳定性。
验证例2 PFD-TK-Cy纳米颗粒的升温情况
取不同浓度的PFD-TK-Cy纳米颗粒水溶液于1.5ml Ep管中,用激发波长为660nm的激光照射4分钟,通过红外热成像仪实时记录溶液温度;图7为纳米颗粒的升温情况(A)以及在不同升温条件下的红外热成像仪的结果(B);结果显示,单纯的水溶液没有升温,PFD-TK-Cy纳米颗粒浓度越高,升温效果越明显,浓度为50微克每毫升的纳米颗粒在4分钟温度可升至46摄氏度,满足温和光热的条件。
验证例3 PFD-TK-Cy纳米颗粒对细胞毒性的影响
实施例1制得的PFD-TK-Cy纳米颗粒对三种细胞毒性的影响,具体来说,将1万个4T1细胞接种于96孔板,在37℃,5%CO2恒温常氧培养箱中进行培养,细胞贴壁后吸去培养基,分别加入100μL含不同浓度的PFD-TK-Cy纳米颗粒的RPMI 1640培养液,孵育24h,采用氮唑溴盐(MTT)法,计算细胞存活率。图8展示的结果发现,PFD-TK-Cy纳米颗粒在24h和48h对肿瘤相关成纤维细胞CAFs的杀伤远高于肿瘤相关成纤维细胞和正常细胞NIH 3T3。
验证例4 PFD-TK-Cy纳米颗粒对肿瘤细胞的杀伤选择性
实施例1制得的PFD-TK-Cy纳米颗粒加光照条件下对肿瘤细胞和肿瘤相关成纤维细胞(CAFs)的杀伤,具体来说,将1万个4T1细胞接种于96孔板,在37℃,5%CO2恒温常氧培养箱中进行培养,细胞贴壁后吸去培养基,分别加入100μL含不同浓度的PFD-TK-Cy纳米颗粒PFD-TK-Cy纳米颗粒的RPMI 1640培养液,孵育8h,对各孔使用激发波长为660nm的激光照射5min,调节功率使得使得温度始终维持在42~43摄氏度之间。继续培养24h后,采用氮唑溴盐(MTT)法,计算细胞存活率。图9结果显示,光照条件下,PFD-TK-Cy纳米颗粒对CAFs的杀伤远高于4T1,说明PFD-TK-Cy纳米颗粒对CAFs有更高的杀伤选择性。
验证例5 PFD-TK-Cy纳米颗粒在小鼠乳腺癌皮下瘤的抗肿瘤药效考察
本验证例利用小鼠4T1乳腺癌皮下瘤模型考察了光照和不光照条件下PFD-TK-Cy纳米颗粒的抗肿瘤效应,并与上市药物Doxil比较。具体步骤如下:
在6周龄,16~18g的雌性BALB/c小鼠右侧皮下接种小鼠乳腺癌4T1细胞悬液,约1×106个细胞,建立小鼠乳腺癌4T1皮下瘤小鼠模型。当皮下瘤体积约100mm3时,将小鼠随机分为5组,每组6只,分别是生理盐水组、PFD-TK-Cy纳米颗粒+光照组、Doxil组、PFD-TK-Cy纳米颗粒+Doxil组、PFD-TK-Cy纳米颗粒+光照+Doxil组。治疗中PFD-TK-Cy纳米颗粒给药剂量为8mg/kg,Doxil给药剂量为2.5mg/kg。记第一天给药时间为第1天,再分别于第5、12天按上述剂量给药,给药1h后,光照组进行光照,光照参数为660nm,10min,保证小鼠肿瘤处温度维持在42℃~43℃之间,给药24h后微静脉注射Doxil。自第1天起,每隔一天测量小鼠体重及瘤体积,绘制瘤体积-时间曲线。在第15天处死小鼠,剥离皮下瘤拍照。图10内容(A)为小鼠瘤体积-时间曲线,内容(B)为剥离后肿瘤照片。结果发现,单纯的PFD-TK-Cy纳米颗粒加光照对肿瘤的抑制效果与单纯的生理盐水组没有差别,说明PFD-TK-Cy纳米颗粒产生的温和光热不会对肿瘤造成杀伤,只是改变了肿瘤的力学环境,Doxil组的肿瘤抑制率为54%,PFD-TK-Cy纳米颗粒+Doxil组肿瘤抑制率为69%,而PFD-TK-Cy纳米颗粒+光照组+Doxil组的肿瘤抑制率高达87%,说明无论是PFD-TK-Cy纳米颗粒还是PFD-TK-Cy纳米颗粒加光照,都可以通过改善肿瘤异常的力学微环境来促进上市药物Doxil在肿瘤部位的蓄积,从而提高抗肿瘤效果。该结果进一步证明了疗效优异的PFD-TK-Cy纳米颗粒在调控肿瘤力学从而改善纳米药物在肿瘤部位递送和分布不足的难题有着巨大的临床潜力。
验证例6 PFD-TK-Cy纳米颗粒的生物安全性
实施例1的PFD-TK-Cy纳米颗粒在小鼠乳腺癌皮下瘤的抗肿瘤药效结束后,对小鼠血液的血常规进行测试用以评价单药组及联合给药方案的安全性。图11为PFD-TK-Cy纳米颗粒的生物安全性研究结果,其中,(A)表示白细胞量,(B)表示红细胞量,(C)表示血小板量,(D)表示血红蛋白量;结果发现,PFD-TK-Cy纳米颗粒处理组的小鼠血常规分析在正常范围,PFD-TK-Cy纳米颗粒具有良好的生物安全性。
实施例1的PFD-TK-Cy纳米颗粒在小鼠乳腺癌皮下瘤的抗肿瘤药效结束后,对小鼠主要器官(心、肝、脾、肺、肾)的组织切片进行安全性评价,图12结果显示,注射实施例2制得的纳米制制剂并没有对小鼠的主要器官(心、肝、脾、肺、肾)造成明显损伤,这表明本实施例制备的的纳米颗粒分散液对正常组织没有造成明显的毒副作用。
按与实施例1相同的步骤也制备了实施例2-实施例18,其结构如表1所示:
表1实施例2-实施例18的整体结构
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其中,除实施例1已经涉及的基质调控类药物和刺激响应的连接分子外,其它基质调控类药物和刺激响应的连接分子的结构如下:
(即将原曲尼斯特中的羧基改性成羟基,不影响其药物活性)
经验证,实施例2-实施例18也具有和实施例1类似的性质。
本领域的技术人员容易理解,以上所述仅为本发明的较佳实施例而已,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、等同替换和改进等,均应包含在本发明的保护范围之内。
Claims (10)
1.一种基质调控类药物-半花菁的偶联物,其特征在于,,所述偶联物的化学结构如式(Ⅰ)所示:
其中,R1为基质调控类药物,R2为刺激响应的连接分子,R3为C1~C18的烷基或烷基磺酸。
2.如权利要求1所述的偶联物,其特征在于,所述基质调控类药物为氯沙坦、曲尼斯特、吡非尼酮或卡泊三醇。
3.如权利要求1所述的偶联物,其特征在于,所述刺激响应的连接分子为ROS响应键、GSH响应键、酶响应键或pH响应键。
4.如权利要求1所述的偶联物,其特征在于,所述偶联物在分散液中以自组装形成的纳米颗粒的形式存在。
5.如权利要求4所述的偶联物,其特征在于,所述分散液中偶联物的浓度为2~20mg/mL。
6.制备如权利要求1-5中任意一项所述偶联物的方法,,其特征在于,所述偶联物的合成路线如下:
7.如权利要求6所述的方法,其特征在于,式(IV)化合物的合成方式如下:
8.如权利要求7所述的方法,其特征在于,式(Ⅲ)化合物的合成方式如下:
9.权利要求1-5中任一项所述的偶联物应用于抗肿瘤药物。
10.如权利要求9所述的应用,其特征在于,所述肿瘤为乳腺癌、卵巢癌、胰腺癌、肝癌、结肠癌或黑色素瘤。
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