CN107802836B - 一种肿瘤靶向光热药剂及制备方法和应用 - Google Patents
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Abstract
本发明公开了一种肿瘤靶向光热药剂及制备方法和应用。本发明通过巯基改性带有靶向基团的PEG对碲化钼进行亲水性改性,合成出具有光热抗肿瘤作用的药剂。此类药剂具有肿瘤细胞靶向的特点,并且可以通过光热作用实现抑制肿瘤细胞的增殖的疗效。
Description
技术领域
本发明属于纳米材料技术领域,具体涉及一种肿瘤靶向光热药剂及制备方法和应用。
背景技术
光热疗法是一种新兴的基于由光激发材料产生远红外波的治疗方法。随着共轭有机基团、碳纳米管、金纳米棒等光热试剂的发展,光热疗法已经广泛的用于肿瘤成像和治疗。光热药剂的特异性和选择性在诊断和治疗的过程中起着关键的作用。纳米材料的几何结构对其光热效果有很大的影响。近年来,具有纳米级厚度的MoS2,MoSe2,WS2,WSe2等过渡性金属双硫属化合物引起了广泛的关注。由于其独特的物理和化学性质,过渡性金属双硫属化合物在电子元器件和光催化和能量储存方面具有很强的应用潜力。受到电化学应用的启发,越来越多的研究者倾向于研究那些具有优异的光学和电子性质的过渡性金属双硫属化合物在癌症成像和治疗方面的应用。例如,在最近的一项光热与化疗联合治疗的研究中刘[ Advanced materials 2014, 26, 3433.]与他的同事们制备了一种MoS2纳米薄片。
二碲化钼是一种灰色六角形粉末状固体。能在碱中分解,不溶于水,可溶于硝酸,在空气中较稳定。可以代替硅成为新型的半导体材料。目前未见将MoTe2用于癌症的多功能疗法中。
发明内容
本发明药解决的第一技术问题是提供一种肿瘤靶向光热药剂的制备方法。该方法第一次将二碲化钼用于癌症的多功能疗法,通过机械球磨法制备了MoTe2纳米薄片,并利用硫醇反应功能化,赋予其肿瘤靶向性与生物相容性。本发明提供了一种生物相容性优良的MoTe2纳米薄片展现出了较强的近红外吸收,低毒性和体内可降解的性质,这使得它们成为光热疗法的一个很有前景的试剂。
本发明要解决的第二个技术问题是提供上述方法制得的肿瘤靶向光热药剂的应用。
为解决上述第一个技术问题,发明一种肿瘤靶向光热药剂的制备方法,步骤如下:
a)将碲化钼晶体在高速球磨机中球磨24-36 h,球磨时旋转速度为200-800 rpm,然后使用声波降解法在冰浴中将球磨后的粉末均匀分散在去离子水中,
b)将分散在水中的碲化钼与巯基改性带有靶向基团的PEG按质量比1:1的比例混合搅拌反应48 h,之后在8000 rpm下离心分离,得到生物相容性优良的MoTe2纳米薄片。
所述的声波降解法为:0-10 oC条件下将球磨后的粉末在细胞粉碎仪的作用下剥离,粉碎时间为6-12 h,得到的纳米片层为1~10层的分子层组成。
优选地,步骤b)中,靶向基团为RGD、cRGD、HA、PAA。
优选地,步骤b)中,PEG的分子量可以是200-5000。
进一步地,本发明提供有上述方法制备的肿瘤靶向光热药剂。
为解决上述第二个技术问题,本发明肿瘤靶向光热药剂在制备抗肿瘤药物中的应用。
如无特殊说明,本发明中的各原料均可通过市售购买获得,本发明中所用的设备可采用所属领域中的常规设备或参照所属领域的现有技术进行。
与现有技术相比较,本发明具有如下有益效果:
本发明选用了高光热转化率的碲化钼纳米材料作为光热试剂,避免了重金属与高毒性芳香类化合物的使用,相对市场同类产品成本低廉,体内代谢快。本发明所述的新型肿瘤靶向光热药剂,对近红外光有强的吸收;对肿瘤细胞的生长具有良好的抑制作用,并且对肿瘤细胞具有更高的杀伤力;有高效的光热转化效率;可以诱导细胞凋亡。
附图说明
图1:碲化钼光热抗肿瘤药剂的TEM图。
图2:抗肿瘤药剂对紫外光的吸收。
图3:a. 抗肿瘤光热药剂对小鼠乳腺癌细胞的生长抑制实验。b. 抗肿瘤光热药剂受远红外辐照后对小鼠乳腺癌细胞的生长抑制实验。
图4:远红外辐照下的抗肿瘤光热药剂对水体系温度的温度分布图。
图5:远红外辐照抗肿瘤光热药剂对体系温度的影响随时间的变化以及水对体系温度的影响比较。
图6:在不同作用条件下癌细胞的凋亡分析。
具体实施方式
通过以下详细说明结合附图可以进一步理解本发明的特点和优点。所提供的实施例仅是对本发明方法的说明,而不以任何方式限制本发明揭示的其余内容。
【实施例1】肿瘤靶向光热药剂1的合成:
(1)将碲化钼晶体在高速球磨机中球磨48 h,转速为400 rmp。
(2)使用细胞粉碎仪将球磨后的粉末均匀分散在去离子水中,粉碎时间为12 h,温度为0~4 oC。
(3)将分散在水中的碲化钼与HS-PEG-cRGD (Mn = 25.7 kg/mol) 按质量比1:1的比例混合搅拌反应48 h,之后在8000 rpm下离心分离,得到生物相容性优良的MoTe2纳米薄片。
(4)用投射电子显微镜表征材料结构(图1),并将获得的产品在-18℃避光条件下保存。
【实施例2】肿瘤靶向光热药剂1的紫外可见吸收光谱
MoTe2纳米薄片水溶液的紫外可见吸收光谱是由UV-Vis分光光度计(ThermoScientific, Waltham, MA)测得。结果如图2所示,MoTe2纳米薄片紫外可见吸收光谱在800~950 nm处显示出强的近红外吸收,从而证明MoTe2纳米薄片是一种有光热转化潜能的化合物,最优吸收波长范围为800~950 nm。
【实施例3】肿瘤靶向光热药剂1作用于小鼠乳腺癌细胞
小鼠乳腺癌细胞以6000个细胞/孔的密度分别接种在2个96孔板中,用0.1毫升培养基培养24小时。然后,将用培养基配制成不同浓度的 MoTe2纳米薄片PBS溶液分别加入到各孔中。在避光条件下保持37摄氏度培养48小时。暗毒性组在避光条件下继续培养24小时。为了评估材料的光毒性,在培养6小时后用新鲜的培养基置换还没有被內吞的材料,然后使用NIR激光进行辐照。每个孔使用808 nm的近红外光,功率为1 W cm-1的激光照射1分钟 。再继续培养24个小时。然后,再在每个孔中加入20微升的 MTT(MTT溶解在5毫克/毫升的PBS缓冲液中)。共培养4小时,吸出培养基,加入150微升二甲亚砜。酶标仪测量每个孔中570纳米处的吸光值,计算细胞存活率,进而得到实施例1所得材料对小鼠乳腺癌细胞的光毒性与暗毒性。
结果如图3所示,光照后的材料对小鼠乳腺癌细胞具有极大的毒性,从而证明光热抗肿瘤化合物对肿瘤细胞具有选择性杀死的特点。
【实施例4】测试亲水的碲化钼纳米薄片的光热性能
为了测试亲水的碲化钼纳米薄片的光热性能,采用808 nm的远红外激光辐照亲水性纳米薄片与空白对照组。用红外热成像仪记录了激光辐照的过程。
结果如图4和图5所示,随着辐照时间的增长,碲化钼薄片组的温度明显提高,空白对照组则几乎没有温度变化。
【实施例5】肿瘤靶向光热药剂1诱导肿瘤细胞凋亡
将小鼠乳腺癌细胞以1×105个细胞/孔的密度接种在六孔板中培养,37℃条件下在2毫升培养基中培养。24小时后,加入溶解有材料的培养基与空白培养基到六孔板中。继续培养6小时后,用808 nm的激光辐照。与不进行任何处理、仅用激光辐照或仅加入材料进行对照。经过进一步培养18小时后,所有的细胞通过胰蛋白酶消化,并离心收集。使用PBS和膜联蛋白缓冲液清洗后,所有细胞都用FITC染色30分钟,通过流式细胞仪进行测试分析。
结果如图6所示,远红外辐照后的材料组呈现出明显的细胞凋亡,这很好的验证了材料具有优异的光热效应。
Claims (5)
1.一种肿瘤靶向光热药剂的制备方法,其特征在于,包括步骤如下:
a)MoTe2纳米薄片的制备:将碲化钼晶体在高速球磨机中球磨24-36 h,球磨时旋转速度为200-800 rpm,然后将球磨后的粉末在细胞粉碎仪的作用下剥离,得到的纳米片层为1~10层的分子层组成,均匀分散在去离子水中,粉碎时间为6-12 h,温度为0-10 oC;
b)肿瘤靶向光热药剂的制备:将分散在水中的MoTe2纳米薄片与巯基改性带有靶向基团的PEG按质量比1:1的比例混合搅拌反应48 h,之后在8000 rpm下离心分离,得到生物相容性优良的MoTe2纳米薄片。
2.根据权利要求1所述的制备方法,其特征在于,步骤b)中,靶向基团为RGD、cRGD、HA、PAA。
3.根据权利要求1所述的制备方法,其特征在于,步骤b)中,PEG的分子量是200-5000。
4.一种肿瘤靶向光热药剂,其特征在于,由权利要求1所述的方法制备获得。
5.权利要求4所述的肿瘤靶向光热药剂在制备抗肿瘤药物中的应用。
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