CN113082228A - 一种基于单核多模态成像用于诊断乳腺癌的造影剂 - Google Patents
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Abstract
本发明涉及一种基于单核多模态成像用于诊断乳腺癌的造影剂材料,其特征在于单核由Li+、Ce3+、Eu3+、Er3+离子掺杂MnO组成,粒径为10‑20nm,经靶向uMUC‑1抗原的多肽Glu‑Pro‑Pro‑Thr(EPPT)修饰,该造影剂具有良好的生物相容性、主动靶向乳腺癌细胞、pH敏感磁共振成像、荧光成像等特性,其材料结构组成是(MnO,0~1% Li+,0~1% Ce3+,0~1% Eu3+,0~1% Er3+)‑EPPT。该材料在乳腺癌诊断和治疗方面具有良好的应用前景。
Description
技术领域
本发明涉及一种基于单核多模态成像用于诊断乳腺癌的造影剂,属生物医学科学领域。
背景技术
全球每年乳腺癌新发病例高达200多万例,早期发现乳腺癌对于改善治疗结果和患者生存率至关重要。因此快速精准的乳腺癌的检测技术和方法尤为重要,当前乳腺癌的检查主要手段有乳腺钼靶照相、乳腺B超、B超引导乳腺穿刺活检、乳管内视镜检查、X线引导导丝定位活检、磁共振成像、荧光成像等。近年来发展起来的多模态成像技术,融合B超、磁共振成像、荧光成像等多种成像技术,取长补短,做到优势互补,而备受青睐和关注。多模态成像技术的关键是造影剂,造影剂的质量往往决定着肿瘤成像的精准度,目前国内外学者开发了大量基于纳米可控制备技术的造影剂,并取得巨大的进步。但是大多数基于多模态成像的造影剂是将有单个功能的多种成分组合到一个纳米平台,例如将有机荧光染料与具有磁共振成像能力的无机纳米材料结合形成的核壳结构,其具备荧光和磁共振多模态成像等能力。然而这种功能各异的组分复合而成的造影剂,往往由于成分复杂、重复性低、药代动力学不明确等原因,难以进行临床转化。
发明内容
针对现有技术的不足,本发明要解决的问题是基于单核多模态成像的思路,将单一组分材料(即所谓“单核”)经过纳米加工技术处理成为集成荧光和磁共振成像等多功能的平台,实现多模态成像,克服现有多模态成像造影剂的缺点。
本发明涉及一种基于单核多模态成像用于诊断乳腺癌的造影剂。其特征是单核由Li+、Ce3+、Eu3+、Er3+离子掺杂MnO组成,粒径为10-20nm,经靶向uMUC-1抗原的多肽Glu-Pro-Pro-Thr(EPPT)修饰,该造影剂具有良好的生物相容性、主动靶向乳腺癌细胞、pH敏感磁共振成像、荧光成像等特性。本发明所述材料结构组成是(MnO,0~1% Li+,0~1% Ce3+,0~1% Eu3 +,0~1% Er3+)-EPPT,修饰于掺杂MnO纳米颗粒表面的EPPT,可与乳腺癌细胞中过度表达的低糖基化黏蛋白1(uMUC-1)结合,而后纳米颗粒将被肿瘤细胞特异性内吞,肿瘤细胞内体pH值较正常组织细胞低,处于低pH值环境中MnO纳米颗粒会释放Mn2+启动MRI信号;离子掺杂赋予MnO良好的荧光成像能力。
本发明所述的一种基于单核多模态成像用于诊断乳腺癌的造影剂材料,由下述步骤实现:
(1)分别称取10 mmol的乙酰丙酮锰(II)、0~0.05 mmol乙酰丙酮锂、0~0.1mmol乙酰丙酮铒(III)、0~0.1mmol乙酰丙酮化铕(III)、0~0.1mmol乙酰丙酮铈(III)溶于30~60 mL的油胺和5~30mL的二苄基醚混合液中,持续搅拌30~60分钟,搅拌过程中持续通入稳定流量的氮气;
(2)将步骤1中的混合液体置入专用的反应釜中,快速升温至110℃,保温20~30分钟,然后,以15~25℃/min的升温速度加热至300℃,保温20~30分钟后快速冷却至室温,反应全程通入稳定流量的氮气并不停搅拌;
(3)取步骤2的反应液,加入同体积的无水乙醇沉淀,9000 r/min 离心10~15 min,去掉上清液,收集沉淀,重复洗涤3~5次,然后将沉淀产物分散到正己烷中,真空干燥至正己烷挥发完毕,得到粒径为10~20纳米的离子掺杂MnO纳米颗粒;
(4)取浓度为500μg/mL EPPT多肽与20 mg的掺杂MnO混合后孵育60~120分钟,9000r/min 离心洗去未结合多肽,最后将交联形成的靶向乳腺肿瘤细胞的纳米造影剂37℃真空干燥。
具体实施方式
下面结合实施例对本发明作进一步阐述,但本发明保护内容不仅限于所述实施例。
实施例1:
(1)分别称取10mmol的乙酰丙酮锰(II)、0.03 mmol乙酰丙酮锂、0.05 mmol乙酰丙酮铒(III)、0.05 mmol乙酰丙酮化铕(III)、0.05 mmol乙酰丙酮铈(III)溶于60mL的油胺和10mL的二苄基醚混合液中,持续搅拌60分钟,搅拌过程中持续通入稳定流量的氮气;
(2)将步骤1中的混合液体置入专用的反应釜中,快速升温至110℃,保温30分钟,然后,以25℃/min的升温速度加热至300℃,保温20分钟后快速冷却至室温,反应全程通入稳定流量的氮气并不停搅拌;
(3)取步骤2的反应液,加入同体积的无水乙醇沉淀,9000 r/min 离心15 min,去掉上清液,收集沉淀,重复洗涤3次,然后将沉淀产物分散到正己烷中,真空干燥至正己烷挥发完毕,得到粒径为10~20纳米的MnO掺杂纳米颗粒;
(4)取浓度为500μg/mL EPPT多肽与20 mg的掺杂MnO混合后孵育90分钟,9000 r/min 离心洗去未结合的多肽,最后将交联形成靶向乳腺肿瘤细胞的纳米造影剂37℃真空干燥。
实施例2:
(1)分别称取10 mmol的乙酰丙酮锰(II)、0.01 mmol乙酰丙酮锂、0.01 mmol乙酰丙酮铒(III)、0.01 mmol乙酰丙酮化铕(III)、0.01 mmol乙酰丙酮铈(III)溶于60mL的油胺和10mL的二苄基醚混合液中,持续搅拌30分钟,搅拌过程中持续通入稳定流量的氮气;
(2)将步骤1中的混合液体置入专用的反应釜中,快速升温至110℃,保温20分钟,然后,以25℃/min的升温速度加热至300℃,保温20分钟后快速冷却至室温,反应全程通入稳定流量的氮气并不停搅拌;
(3)取步骤2的反应液,加入同体积的无水乙醇沉淀,9000 r/min 离心10min,去掉上清液,收集沉淀,重复洗涤3次,然后将沉淀产物分散到正己烷中,真空干燥至正己烷挥发完毕,得到粒径为10~20纳米的MnO掺杂纳米颗粒;
(4)取浓度为500μg/mL EPPT多肽与20 mg的掺杂MnO混合后孵育60分钟,9000 r/min 离心洗去未结合的多肽,最后将交联形成靶向乳腺肿瘤细胞的纳米造影剂37℃真空干燥。
实施例3:
(1)分别称取10mmol的乙酰丙酮锰(II)、0.02 mmol乙酰丙酮锂、0.01 mmol乙酰丙酮铒(III)、0.1 mmol乙酰丙酮化铕(III)、0.01 mmol乙酰丙酮铈(III)溶于60mL的油胺和10mL的二苄基醚混合液中,持续搅拌60分钟,搅拌过程中持续通入稳定流量的氮气;
(2)将步骤1中的混合液体置入专用的反应釜中,快速升温至110℃,保温30分钟,然后,以25℃/min的升温速度加热至300℃,保温30分钟后快速冷却至室温,反应全程通入稳定流量的氮气并不停搅拌;
(3)取步骤2的反应液,加入同体积的无水乙醇沉淀,9000 r/min 离心15 min,去掉上清液,收集沉淀,重复洗涤3次,然后将沉淀产物分散到正己烷中,真空干燥至正己烷挥发完毕,得到粒径为10~20纳米的MnO掺杂纳米颗粒;
(4)取浓度为500μg/mL EPPT多肽与20 mg的掺杂MnO混合后孵育120分钟,9000 r/min 离心洗去未结合多肽,最后将交联形成靶向乳腺肿瘤细胞的纳米造影剂37℃真空干燥。
Claims (1)
1.一种一种基于单核多模态成像用于诊断乳腺癌的造影剂,其特征是:单核由Li+、Ce3 +、Eu3+、Er3+离子掺杂MnO组成,粒径为10-20nm,经靶向uMUC-1抗原的多肽Glu-Pro-Pro-Thr(EPPT)修饰,其中材料结构组成是(MnO,0~1% Li+,0~1% Ce3+,0~1% Eu3+,0~1% Er3+)-EPPT,该基于单核多模态成像用于诊断乳腺癌的造影剂材料材料由如下步骤实现制备:
(1)分别称取10 mmol的乙酰丙酮锰(II)、0~0.05 mmol乙酰丙酮锂、0~0.1mmol乙酰丙酮铒(III)、0~0.1mmol乙酰丙酮化铕(III)、0~0.1mmol乙酰丙酮铈(III)溶于30~60 mL的油胺和5~30mL的二苄基醚混合液中,持续搅拌30~60分钟,搅拌过程中持续通入稳定流量的氮气;
(2)将步骤1中的混合液体置入专用的反应釜中,快速升温至110℃,保温20~30分钟,然后,以15~25℃/min的升温速度加热至300℃,保温20~30分钟后快速冷却至室温,反应全程通入稳定流量的氮气并不停搅拌;
(3)取步骤2的反应液,加入同体积的无水乙醇沉淀,9000 r/min 离心10~15 min,去掉上清液,收集沉淀,重复洗涤3~5次,然后将沉淀产物分散到正己烷中,真空干燥至正己烷挥发完毕,得到粒径为10~20纳米的离子掺杂MnO纳米颗粒;
(4)取浓度为500μg/mL EPPT多肽与20 mg的掺杂MnO混合后孵育60~120分钟,9000 r/min 离心洗去未结合多肽,最后将交联形成的靶向乳腺肿瘤细胞的纳米造影剂37℃真空干燥。
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CN107550864A (zh) * | 2017-08-23 | 2018-01-09 | 上海市计划生育科学研究所 | Eppt多肽‑聚乙二醇‑磷脂复合膜材料、其制备方法及主动靶向脂质体递药系统和应用 |
CN109620972A (zh) * | 2019-01-24 | 2019-04-16 | 广州创赛生物医用材料有限公司 | 一种用于肺癌诊断的t1-t2双模态靶向成像造影剂及制备方法 |
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CN104043138A (zh) * | 2014-05-29 | 2014-09-17 | 北京大学 | 稀土基纳米颗粒磁共振造影剂及其制备方法 |
CN107550864A (zh) * | 2017-08-23 | 2018-01-09 | 上海市计划生育科学研究所 | Eppt多肽‑聚乙二醇‑磷脂复合膜材料、其制备方法及主动靶向脂质体递药系统和应用 |
CN109620972A (zh) * | 2019-01-24 | 2019-04-16 | 广州创赛生物医用材料有限公司 | 一种用于肺癌诊断的t1-t2双模态靶向成像造影剂及制备方法 |
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