CN106362172B - 透明质酸修饰的介孔碳酸钙药物组合物的制备方法及应用 - Google Patents
透明质酸修饰的介孔碳酸钙药物组合物的制备方法及应用 Download PDFInfo
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Abstract
本发明涉及透明质酸修饰的介孔碳酸钙药物组合物的制备方法及应用,可有效解决同时具有靶向、控释、声疗联合超声成像的问题,实现癌症诊疗一体化,技术方案是:该组合物是在介孔碳酸钙与3‑氨丙基三乙氧基硅烷反应得到的氨基化的碳酸钙上修饰上透明质酸,再负载小分子声敏剂,得透明质酸修饰的介孔碳酸钙药物组合物;本发明制备工艺简单,方法稳定可靠,生产费用低,制备的透明质酸修饰的介孔碳酸钙及其药物组合物同时具备肿瘤靶向、体内外双刺激(酸性环境和高聚焦超声条件下)敏感释药、声动力学治疗联合超声成像实现诊疗一体化等多重功效,是肿瘤治疗药物上的创新,经济和社会效益巨大。
Description
技术领域
本发明涉及医药领域,特别是透明质酸修饰的介孔碳酸钙药物组合物的制备方法及应用。
背景技术
目前,超声成像技术以无创伤、无辐射、价廉、动态实时以及诊断参数多样和工程上灵活等优点,已被广泛应用于临床医学领域,成为疾病医学检查与影像诊断的有效手段。为了提高普通超声图像的分辨率及对比度,需要引入了一类能够显著增强医学超声检测信号的试剂——超声造影剂。介孔碳酸钙纳米粒(MCC)作为药物储库,其独特的介孔结构可以负载小分子药物,具有极高的载药能力。另外,在肿瘤弱酸性环境(pH 5.6)下,MCC可与大量的H+反应崩解并缓慢的产生二氧化碳气泡,当无外界刺激时,这些二氧化碳气体藏匿在介孔碳酸钙的孔洞中,而在外加超声作用下,介孔孔洞内的气泡膨胀(随着温度升高气体压强变大,气泡体积随之变大),使得MCC更易崩解,瞬间释放药物,进行超声成像。因此,介孔碳酸钙作为一种新型的超声造影剂,产生的二氧化碳气泡可大幅度增强血流回波强度,极大提高了超声图像中灌注组织的对比度,弥补了原有普通超声检测信号的不足。
近年来,声动力疗法(sonodynamic therapy,SDT)已发展成为继手术、放疗、化疗之后肿瘤治疗新技术。SDT 是指临床给予患者一定剂量声敏剂(sonosensitizer),使用高强度聚焦超声(High Intensity Focused Ultrasound,HIFU)辐照肿瘤部位,激发声敏剂产生活性氧,促使肿瘤细胞发生不可逆性损伤,从而达到肿瘤治疗的目的。但对于小分子声敏剂的应用目前仍有一些限制,如肿瘤靶向性低,水分散性差等问题。
发明内容
针对上述情况,为克服现有技术之缺陷,本发明之目的就是提供透明质酸修饰的介孔碳酸钙药物组合物的制备方法及应用,可有效解决同时具有靶向、控释、声疗联合超声成像的问题,实现癌症诊疗一体化。
本发明解决的技术方案是,该组合物是在介孔碳酸钙与3-氨丙基三乙氧基硅烷反应得到的氨基化的碳酸钙上修饰上透明质酸,再负载小分子声敏剂,得透明质酸修饰的介孔碳酸钙药物组合物;所述的透明质酸为分子量等于或低于400 kDa,并等于或高于40kDa的低分子量透明质酸;所述的介孔碳酸钙的粒径为300~400nm;所述小分子声敏剂为二氢卟吩、金属酞菁及血卟啉单甲醚中的一种。
所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,包括以下步骤:
(1)将0.6-0.7g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.6-0.7g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2-3g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液100-150ml 和可溶性淀粉溶液100-150ml混合,搅拌20-40min,再向该混合液中加入100-150ml Na2CO3溶液,剧烈搅拌10-20min,静置22-26h;12000rpm离心5-10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,得介孔碳酸钙纳米颗粒(MCC);
(3)将140-200mg介孔碳酸钙纳米颗粒超声分散于60-80ml无水乙醇中,再加入600-800μl 3-氨丙基三乙氧基硅烷,3-4ml水和3-4ml浓氨水,搅拌8-12h,12000rpm离心5-10min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物(MCC-NH2);
(4)将700-800mg透明质酸加入pH 7-9的磷酸盐缓冲液40-50ml中,搅拌溶解,加入1.8-2.0g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.0-1.2g活化剂羟基琥珀酰亚胺,室温搅拌25-35min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH7-9的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应2.5-3.5h,12000rpm离心5-10min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物(MCC-HA);
(6)将12-18mg小分子声敏剂溶解于4-6ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物4-6mg探超分散于4-6ml超纯水中,搅拌20-30h,12000rpm离心5-10min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物;所述小分子声敏剂为二氢卟吩、金属酞菁及血卟啉单甲醚中的一种。
所述的透明质酸修饰的介孔碳酸钙药物组合物在制备抗肿瘤药物中的应用。
所述的透明质酸修饰的介孔碳酸钙药物组合物在制备治疗肿瘤药物注射剂、口服剂或植入给药剂中的应用。
所述的透明质酸修饰的介孔碳酸钙药物组合物在制备肿瘤靶向、体内外双重敏感释药、声疗联合超声成像药物中的应用。
本发明制备工艺简单,方法稳定可靠,生产费用低,制备的透明质酸修饰的介孔碳酸钙及其药物组合物同时具备肿瘤靶向、体内外双刺激(酸性环境和高聚焦超声条件下)敏感释药、声动力学治疗联合超声成像实现诊疗一体化等多重功效,是肿瘤治疗药物上的创新,经济和社会效益巨大。
具体实施方案
下以下实施例对本发明的具体实施方式作详细说明。
实施例1
本发明所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,包括以下步骤:
(1)将0.66g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.63g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2.5g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液125ml 和可溶性淀粉溶液125ml混合,搅拌30min,再向该混合液中加入125ml Na2CO3溶液,剧烈搅拌15min,静置24h;12000rpm离心5-10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2遍,冷冻干燥,得介孔碳酸钙纳米颗粒MCC;
(3)将170mg介孔碳酸钙纳米颗粒超声分散于70ml无水乙醇中,再加入700μl 3-氨丙基三乙氧基硅烷,3.5ml水和3.5ml浓氨水,搅拌10h,12000rpm离心8min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物MCC-NH2;
(4)将750mg透明质酸加入pH 7-9的磷酸盐缓冲液45ml中,搅拌溶解,加入1.9g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.1g活化剂羟基琥珀酰亚胺,室温搅拌30min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH8的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应3h,12000rpm离心8min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物MCC-HA;
(6)将15mg小分子声敏剂二氢卟吩溶解于5ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物5mg探超分散于5ml超纯水中,搅拌24h,12000rpm离心8min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
实施例2
本发明所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,包括以下步骤:
(1)将0.6g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.6g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液100ml 和可溶性淀粉溶液100ml混合,搅拌20min,再向该混合液中加入100ml Na2CO3溶液,剧烈搅拌10min,静置22h;12000rpm离心5min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2遍,冷冻干燥,得介孔碳酸钙纳米颗粒MCC;
(3)将140mg介孔碳酸钙纳米颗粒超声分散于60ml无水乙醇中,再加入600μl 3-氨丙基三乙氧基硅烷,3ml水和3ml浓氨水,搅拌8h,12000rpm离心5min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物MCC-NH2;
(4)将700mg透明质酸加入pH 7的磷酸盐缓冲液40ml中,搅拌溶解,加入1.8g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.0g活化剂羟基琥珀酰亚胺,室温搅拌25min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH7的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应2.5h,12000rpm离心5min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物MCC-HA;
(6)将12mg小分子声敏剂金属酞菁溶解于4ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物4mg探超分散于4ml超纯水中,搅拌20h,12000rpm离心5min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
实施例3
本发明所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,包括以下步骤:
(1)将0.7g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.7g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉3g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液150ml 和可溶性淀粉溶液150ml混合,搅拌40min,再向该混合液中加入150ml Na2CO3溶液,剧烈搅拌20min,静置26h;12000rpm离心10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复3遍,冷冻干燥,得介孔碳酸钙纳米颗粒MCC;
(3)将200mg介孔碳酸钙纳米颗粒超声分散于80ml无水乙醇中,再加入800μl 3-氨丙基三乙氧基硅烷,4ml水和4ml浓氨水,搅拌12h,12000rpm离心10min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复3遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物MCC-NH2;
(4)将800mg透明质酸加入pH9的磷酸盐缓冲液50ml中,搅拌溶解,加入2.0g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.2g活化剂羟基琥珀酰亚胺,室温搅拌35min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH9的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应3.5h,12000rpm离心10min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物MCC-HA;
(6)将18mg小分子声敏剂血卟啉单甲醚溶解于6ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物6mg探超分散于6ml超纯水中,搅拌30h,12000rpm离心10min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复3遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
本发明选择将小分子声敏剂负载到介孔碳酸钙纳米粒内部,在碳酸钙表面修饰上具有良好生物相容性、肿瘤细胞靶向性的天然多糖——透明质酸(HA),构建了一种新型的纳米载体。该载体的合成工艺简单,并将碳酸钙的超声成像、强大的药物负载特性,透明质酸独特的肿瘤细胞靶向性、良好的生物相容性有机的整合于一体,加之可以物理负载小分子声敏剂,该系统在高聚焦超声条件下既可以成像,实时监测肿瘤组织,所负载的声敏剂又可以产生活性氧杀死肿瘤细胞,实现声动力学治疗。因此,研究一种可以将声动力学治疗与超声成像结合的药物转运系统在药物的控释与癌症治疗方面仍具有重要的意义。
经科学试验,本发明所制得的透明质酸修饰的中空介孔碳酸钙复合物可有效靶向至肿瘤部位,在肿瘤弱酸性环境(pH 5.6)下,并加以高聚焦超声(HIFU)辅助可瞬间崩解产生二氧化碳气泡,同时实现体内外双重刺激敏感释药。所制得的复合物同时具备靶向、控释、声疗联合化疗等多重功效,操作方便,方法稳定可靠,与传统化疗相比具有高效、可控的优势,并且,其声动力学治疗与超声成像技术的结合更体现癌症诊疗一体化的问题。有关资料如下:
一、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的制备和表征
1、透明质酸修饰的介孔碳酸钙纳米复合物中血卟啉单甲醚(HMME/MCC-HA)含量的测定
采用紫外分光光度法,于398nm波长处测定血卟啉单甲醚的含量。以公式(1)计算样品的载药量。载药量达到40%左右。
(1)
2、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的粒径和电位的测定
取适量负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物分散于水中,用Nano-ZS90 型激光纳米粒度分析仪测得其粒径和电位分别为335nm和15.2 ±2.1 mV。
二、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的药物释放实验
取4份HMME/MCC-HA纳米复合物,分设超声组与非超声组,分别分散于50ml pH7.4和 pH5.5含0.5%SDS的磷酸盐缓冲液分散介质中,放置于摇床中(37℃,100rpm),分别于2h,4h,6h,8h,10h,12h,24h取样1ml,之后再加入1ml的释放介质,超声组在每个时间点用功率1W/cm2 超声1min。用高效液相检测样品,释药24h后,pH5.5 和pH5.5+HIFU两组的累积释药量分别为82.4%和93.7%,pH7.4和pH7.4+HIFU两组的累积释药量分别为26.7%和34.2%,酸性环境下的累积释药量远高于中性环境。该结果说明MCC在酸性环境下(pH 5.5)更易崩解,同时在超声辅助下可加剧MCC的崩解,达到酸性环境和高聚焦超声条件下的双重刺激敏感释药的目的。
三、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的细胞增殖抑制实验
采用SRB法,选择对数生长期的MCF-7人乳腺癌细胞,调整细胞数为5×104/ml接种于96孔培养板,每孔100μ1(边缘孔用无菌PBS填充),细胞贴壁生长24h后加药,依次为空白组、HMME组、HMME+HIFU组、HMME/MCC-HA组及HMME/MCC-HA+HIFU组。HMME终浓度设为10μg/ml,并分设超声组(1W/cm2,1min)和非超声组。每组设6个复孔。加药孵育24 h后,每孔加入50μl 4℃预冷的50%三氯乙酸(TCA)固定细胞,固定10min后移入4℃冰箱固定1h,取出弃去固定液,用去离子水洗5遍,甩干,室温自然干燥。室温晾干后,每孔加入SRB染液50μ1,室温避光放置15~30min染色,弃染液,用1%的冰醋酸洗5遍,室温干燥。之后,用150μl非缓冲Tris碱液(10mM,pH=10.5)溶解与细胞蛋白结合的染料,摇床微振荡(37℃,100rpm,10min),于酶标仪515 nm波长处测每个小孔的OD值,计算肿瘤细胞生长抑制率(% )= (1-实验组OD值/对照组OD值)×100%,计算得到HMME组、HMME+HIFU组、HMME/MCC-HA组,HMME/MCC-HA+HIFU组各组的细胞生长抑制率分别为: 5.3%,45.4%,9.7%,82.6%。结果表明,HMME在未超声时无明显细胞毒效应,但在超声条件下产生活性氧可抑制肿瘤生长,有明显的声动力学效应;在无超声时制剂组与原料药组的细胞毒性差别不大,而在超声时HMME/MCC-HA对细胞杀伤效果最强,则是在靶头HA辅助下制剂可被细胞更多的摄取,并在HIFU下释放药物发挥声疗作用。
四、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的药效学研究
购买昆明小鼠(雌性,3~4周龄),在小鼠的右上肢背部皮下接种S-180腹水瘤细胞,7天后测量肿瘤体积,取36只肿瘤体积≥100 mm3 且肿瘤体积和体重相似的小鼠,将其随机分为6组,每组6只。具体分组如下:生理盐水组、HMME组、HMME+HIFU组、MCC-HA组、HMME/MCC-HA组,HMME/MCC-HA+HIFU组。超声组使用的功率为1W/cm2,给药3h后超声治疗肿瘤部位,一次超声时间为1min,6组小鼠的给药方式均采用尾静脉注射,每两天给药一次,共给药7 次。整个实验过程中保证小鼠每日正常饮食,每两天对小鼠进行称重,并使用数显游标卡尺测量荷瘤小鼠肉瘤的长径(A)与短径(B),按公式肿瘤体积V=A×B2/2计算肿瘤体积。记录的数据显示,HMME组、HMME+HIFU组、MCC-HA组、HMME/MCC-HA组,HMME/MCC-HA+HIFU组各组的抑瘤率分别为4.85%, 39.36%, 5.58%,9.21%, 88.67%。结果表明, HMME/MCC-HA+HIFU组的药效显著,说明HMME/MCC-HA肿瘤靶向给药系统在超声下的声动力学疗法可以显著增强肿瘤的治疗效果。
五、负载血卟啉单甲醚的透明质酸修饰的介孔碳酸钙纳米复合物的超声成像实验
在小鼠的右上肢背部皮下接种S-180腹水瘤细胞,7天后测量肿瘤体积,取9只肿瘤体积≥100 mm3 且肿瘤体积和体重相似的小鼠,将其随机分为3组,每组3只,具体分组如下:生理盐水组、MCC、MCC-HA组。对两组小鼠腹腔注射0.04ml 3%戊巴比妥钠进行麻醉,固定后对两组小鼠均采用静脉给药的方式,注射后30min、60min、90min、120min对小鼠肿瘤部位进行超声成像。结果显示MCC-HA组在给药60min后在肿瘤部位有明显的超声信号,MCC组的超声信号较弱,这是由于MCC-HA具有较强的肿瘤靶向性且可在肿瘤酸性部位及超声辅助下产生大量二氧化碳,超声信号明显增强。该结果表明MCC-HA作为新型超声造影剂具有良好的超声成像效果。
实验表明,本发明与现有技术相比,具有以下突出的有益技术效果:
(1)本发明提供的透明质酸修饰的介孔碳酸钙及其药物组合物,在肿瘤弱酸性环境(pH 5.6)下,并加以高聚焦超声(HIFU)辅助可瞬间崩解,实现体内外双重刺激敏感释药。
(2)本发明提供的透明质酸修饰的介孔碳酸钙及其药物组合物结构较为简单,具有优良的生物相容性、水分散性和稳定性,还能够实现肿瘤特异性靶向,并保留声敏剂的声动力学治疗活性;
(3)本发明提供的透明质酸修饰的介孔碳酸钙及其药物组合物,在肿瘤弱酸性环境及HIFU辅助下产生大量二氧化碳气泡,故可作为优良的超声造影剂,同时实现声疗及超声成像结合的诊疗一体化。
Claims (5)
1.一种透明质酸修饰的介孔碳酸钙药物组合物的制备方法,其特征在于,该组合物是在介孔碳酸钙与3-氨丙基三乙氧基硅烷反应得到的氨基化的碳酸钙上修饰上透明质酸,再负载小分子声敏剂,得透明质酸修饰的介孔碳酸钙药物组合物;所述的透明质酸为分子量等于或低于400 kDa,并等于或高于40kDa的低分子量透明质酸;所述的介孔碳酸钙的粒径为300~400nm;所述小分子声敏剂为二氢卟吩、金属酞菁及血卟啉单甲醚中的一种;具体制备方法包括以下步骤:
(1)将0.6-0.7g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.6-0.7g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2-3g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液100-150ml 和可溶性淀粉溶液100-150ml混合,搅拌20-40min,再向该混合液中加入100-150ml Na2CO3溶液,剧烈搅拌10-20min,静置22-26h;12000rpm离心5-10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,得介孔碳酸钙纳米颗粒;
(3)将140-200mg介孔碳酸钙纳米颗粒超声分散于60-80ml无水乙醇中,再加入600-800μl 3-氨丙基三乙氧基硅烷,3-4ml水和3-4ml浓氨水,搅拌8-12h,12000rpm离心5-10min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物;
(4)将700-800mg透明质酸加入pH 7-9的磷酸盐缓冲液40-50ml中,搅拌溶解,加入1.8-2.0g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.0-1.2g活化剂羟基琥珀酰亚胺,室温搅拌25-35min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH7-9的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应2.5-3.5h,12000rpm离心5-10min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物;
(6)将12-18mg小分子声敏剂溶解于4-6ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物4-6mg探超分散于4-6ml超纯水中,搅拌20-30h,12000rpm离心5-10min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2-3遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物;所述小分子声敏剂为二氢卟吩、金属酞菁及血卟啉单甲醚中的一种。
2.根据权利要求1所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,其特征在于,包括以下步骤:
(1)将0.66g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.63g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2.5g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液125ml 和可溶性淀粉溶液125ml混合,搅拌30min,再向该混合液中加入125ml Na2CO3溶液,剧烈搅拌15min,静置24h;12000rpm离心5-10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2遍,冷冻干燥,得介孔碳酸钙纳米颗粒;
(3)将170mg介孔碳酸钙纳米颗粒超声分散于70ml无水乙醇中,再加入700μl 3-氨丙基三乙氧基硅烷,3.5ml水和3.5ml浓氨水,搅拌10h,12000rpm离心8min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物;
(4)将750mg透明质酸加入pH 7-9的磷酸盐缓冲液45ml中,搅拌溶解,加入1.9g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.1g活化剂羟基琥珀酰亚胺,室温搅拌30min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH8的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应3h,12000rpm离心8min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物;
(6)将15mg小分子声敏剂二氢卟吩溶解于5ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物5mg探超分散于5ml超纯水中,搅拌24h,12000rpm离心8min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
3.根据权利要求1所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,其特征在于,包括以下步骤:
(1)将0.6g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.6g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉2g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液100ml 和可溶性淀粉溶液100ml混合,搅拌20min,再向该混合液中加入100ml Na2CO3溶液,剧烈搅拌10min,静置22h;12000rpm离心5min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复2遍,冷冻干燥,得介孔碳酸钙纳米颗粒;
(3)将140mg介孔碳酸钙纳米颗粒超声分散于60ml无水乙醇中,再加入600μl 3-氨丙基三乙氧基硅烷,3ml水和3ml浓氨水,搅拌8h,12000rpm离心5min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复2遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物;
(4)将700mg透明质酸加入pH 7的磷酸盐缓冲液40ml中,搅拌溶解,加入1.8g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.0g活化剂羟基琥珀酰亚胺,室温搅拌25min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH7的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应2.5h,12000rpm离心5min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物;
(6)将12mg小分子声敏剂金属酞菁溶解于4ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物4mg探超分散于4ml超纯水中,搅拌20h,12000rpm离心5min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复2遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
4.根据权利要求1所述的透明质酸修饰的介孔碳酸钙药物组合物的制备方法,其特征在于,包括以下步骤:
(1)将0.7g的CaCl2溶解于1000ml水中,配置成CaCl2溶液;将0.7g的Na2CO3溶解于1000ml水中,配置成Na2CO3溶液;将可溶性淀粉3g溶解于1000ml水中,配置成可溶性淀粉溶液;
(2)将步骤(1)配置的CaCl2溶液150ml 和可溶性淀粉溶液150ml混合,搅拌40min,再向该混合液中加入150ml Na2CO3溶液,剧烈搅拌20min,静置26h;12000rpm离心10min得沉淀,沉淀用超纯水复溶,再离心再复溶,如此重复3遍,冷冻干燥,得介孔碳酸钙纳米颗粒;
(3)将200mg介孔碳酸钙纳米颗粒超声分散于80ml无水乙醇中,再加入800μl 3-氨丙基三乙氧基硅烷,4ml水和4ml浓氨水,搅拌12h,12000rpm离心10min得沉淀,沉淀再用无水乙醇复溶,再离心再复溶,如此重复3遍,冷冻干燥,得氨基化介孔碳酸钙纳米复合物;
(4)将800mg透明质酸加入pH9的磷酸盐缓冲液50ml中,搅拌溶解,加入2.0g 1-乙基-(3-二甲基氨基丙基)碳二亚胺、1.2g活化剂羟基琥珀酰亚胺,室温搅拌35min,得活化的透明质酸;
(5)在冰浴下,将步骤(3)制备的氨基化介孔碳酸钙纳米复合物用pH9的磷酸盐缓冲液分散,成1mg/ml的分散液,将活化的透明质酸缓慢加入分散液中,升至室温,反应3.5h,12000rpm离心10min得沉淀,沉淀加水分散,在水中透析2d,冷冻干燥,得透明质酸修饰的介孔碳酸钙纳米复合物;
(6)将18mg小分子声敏剂血卟啉单甲醚溶解于6ml无水乙醇中,搅拌,将步骤(5)制备的透明质酸修饰的介孔碳酸钙纳米复合物6mg探超分散于6ml超纯水中,搅拌30h,12000rpm离心10min得沉淀,沉淀再用无水乙醇和水按照体积比1:1混合的混合液复溶,再离心再复溶,如此重复3遍,冷冻干燥,即得透明质酸修饰的介孔碳酸钙药物组合物。
5.权利要求1所述方法制备的透明质酸修饰的介孔碳酸钙药物组合物在制备肿瘤靶向、体内外双重敏感释药或声疗联合超声成像药物中的应用。
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CN114539684A (zh) * | 2022-02-23 | 2022-05-27 | 江苏华兰药用新材料股份有限公司 | 一种用于注射液药品的卤化丁基橡胶材料及其制备方法 |
CN114948904A (zh) * | 2022-07-28 | 2022-08-30 | 中国中医科学院中药研究所 | 透明质酸衍生物包被的磷酸钙纳米粒的制备方法与应用 |
-
2016
- 2016-08-26 CN CN201610732073.9A patent/CN106362172B/zh active Active
Non-Patent Citations (3)
Title |
---|
CO2 bubbling-based ‘Nanobomb’ System for Targetedly Suppressing Panc-1 Pancreatic Tumor via Low Intensity Ultrasound-activated Inertial Cavitation;Kun Zhang et al.;《Theranostics》;20150912;第5卷(第11期);第1291-1302页 |
Photosensitizer-loaded bubble-generating mineralized nanoparticles for ultrasound imaging and photodynamic therapy;Dong Jin Park et al.;《J. Mater. Chem. B》;20160108;第4卷;第1219-1227页 |
Preparation of hierarchical mesoporous CaCO3 by a facile binary solvent approach as anticancer drug carrier for etoposide;Haibao Peng et al.;《Nanoscale Research Letters》;20130715;第8卷(第321期);第1-11页 |
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