CN114736299A - 一种核素标记的曲妥珠单抗及其制备方法 - Google Patents
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Abstract
本发明公开了一种核素标记的曲妥珠单抗及其制备方法,为放射性核素131I标记的曲妥珠单抗,通过称取Iodogen于试管中并用二氯甲烷溶解,取10μL的Iodogen加到反应管底部,用氮气吹干,使反应管底部形成均匀的薄膜;向涂有一定量Iodogen的反应管中依次加入曲妥珠单抗10.0μg,Na131I溶液10μL,轻轻摇动,室温条件下反应8‑10min;反应完毕后,向反应管中加入250μL PB液,静置,得到反应液;将反应液用SephadexG‑50柱进行纯化,得到目标产物。本发明所述的一种核素标记的曲妥珠单抗及其制备方法,使用质子核素131I,既保证了能在局部产生足够的电离辐射生物学效应,达到抑制及破坏病变组织的目的,又避免对邻近正常组织造成不必要的辐射,实现靶向肿瘤药物的个体化、精准化治疗。
Description
技术领域
本发明涉及临床核医疗技术领域,特别涉及一种核素标记的曲妥珠单抗及其制备方法。
背景技术
随着核医学与分子生物学的深入发展与融合,医学影像技术迈向分子影像学时代,其中正电子发射计算机断层显像(Positron Emission Tomography,PET)进行功能学显像使得人们真正从分子水平开始认识并诊断疾病,尤其在肿瘤的诊疗方面凸显出其优势。通过示踪病变组织的受体变化、细胞信号转导的异常,从而克服现代诊断技术中的缺陷,为肿瘤的早期诊断、临床分期、疗效评估提供依据,并对预后进行评价,同时也可应用于肿瘤的靶向治疗。除了有高分辨率、高灵敏度和高采集速度的多模态成像设备外,还需要高特异性分子探针。分子探针的品质决定了分子影像的质量。敏感、准确、特异性的肿瘤靶向探针是影像学技术的关键。
目前分子核医学中代谢显像在临床中应用最广泛,其中最多见的为18F-FDG,但因其为非特异性显像剂,在HER2阳性肿瘤诊断方面存在很多不足,因此亟需研发特异性显像剂对HER2阳性肿瘤进行显像,以真正发挥PET显像的优势。当然无论肿瘤的靶向显像还是治疗,都依托于放射性药物的研制与合成。若肿瘤中某种受体高水平表达,则可利用受体与配体高特异性、高亲和性结合的特点,将配体进行放射性核素标记,从而作为示踪剂与受体高密度表达部位相结合从而对肿瘤部位显像。
以人表皮生长因子受体2(Human epithelial growth factor receptor 2,HER2)为靶点的人源化单克隆抗体曲妥珠单抗(商品名Trastuzumab)联合化学治疗对HER2过表达的肿瘤较单纯化疗明显延长总生存期(Overall survival,OS)。Trastuzumab单抗能够靶向结合胃癌在内恶性肿瘤细胞膜表面HER2受体,抑制下游相关信号通路激活,进而抑制肿瘤细胞生长及侵袭转移。同时,Trastuzumab结合HER2受体可以启动机体免疫系统抗体依赖的细胞介导的细胞毒性作用进行肿瘤细胞杀伤,给肿瘤细胞以致命一击。文献报道胃癌Her-2蛋白的过表达率为9%-38%。ToGA结果显示,HER2过表达晚期胃癌患者抗HER2靶向治疗客观有效率仅为47.3%。胃癌存在肿瘤异质性,这可能是导致检测HER2状态偏差的主要原因,也可能是IHC(免疫组化)与FISH(荧光原位杂交技术)结果、内镜活检标本和外科手术标本、转移淋巴结结果不一致的主要原因。故此,我们提出了一种核素标记的曲妥珠单抗及其制备方法。
发明内容
本发明的主要目的在于提供一种核素标记的曲妥珠单抗及其制备方法,可以有效解决背景技术中的问题。
为实现上述目的,本发明采取的技术方案为:
一种核素标记的曲妥珠单抗,其为放射性核素131I标记的曲妥珠单抗。
一种核素标记的曲妥珠单抗的制备方法,包括以下步骤:
S1:称取Iodogen于试管中并用二氯甲烷溶解,取10μL的Iodogen加到反应管底部,用氮气吹干,使反应管底部形成均匀的薄膜;
S2:向涂有一定量Iodogen的反应管中依次加入曲妥珠单抗10.0μg,Na131I溶液10μL,轻轻摇动,室温条件下反应8-10min;
S3:反应完毕后,向S2的反应管中加入250μL PB液,静置,得到反应液;
S4:将S3所得的反应液用SephadexG-50柱进行纯化,得到目标产物。
优选的,当标记率大于92%,用SephadexG-50柱纯化后,使目标产物的放射性化学纯度大于98%。
优选的,在所述S1中,所述二氯甲烷的浓度为1g/L。
优选的,在所述S2中,所述Na131I溶液的放射性活度为22.2MBq。
优选的,在所述S3中,所述PB液的浓度为0.05mol/L,静置时间为5-8min。
优选的,在所述S4中,所述SephadexG-50柱使用前,先用0.01MPBS溶液平衡柱体,然后再用0.01M的PBS液纯化出目标产物。
优选的,所述平衡柱体的PBS溶液的量浓度为0.01M/L,pH为7.4;所述纯化目标产物的PBS溶液的量浓度为0.03M/L,pH为7.5。
优选的,所述PBS溶液平衡柱体的具体方法为:每次加5mLPBS溶液,通过重力流速流干,重复3-5次。
与现有技术相比,本发明具有如下有益效果:
1、在本发明使用质子核素131I,131I可同时发射β和γ两种射线,β射线在组织中的射程仅为2-3mm,既保证了能在局部产生足够的电离辐射生物学效应,达到抑制及破坏病变组织的目的,又避免对邻近正常组织造成不必要的辐射,是目前较理想的RIT用放射性核素;
2、在本发明制备的131I—Herceptin在体外对高表达HER-2的人卵巢癌细胞SKOV3有明显的抑制作用,且131I-Herceptin能选择性的长时间浓聚于移植瘤部位。
3、本发明用131I标记Herceptin并作用于转移性胃癌和直肠癌,证实其具有良好的靶向作用,得到了很好的瘤体与正常组织间的吸收比率,实现靶向肿瘤药物的个体化、精准化治疗。
附图说明
图1为本发明一种核素标记的曲妥珠单抗的制备方法的流程图
具体实施方式
为使本发明实现的技术手段、创作特征、达成目的与功效易于明白了解,下面结合具体实施方式,进一步阐述本发明。
在本发明的描述中,需要说明的是,术语“上”、“下”、“内”、“外”“前端”、“后端”、“两端”、“一端”、“另一端”等指示的方位或位置关系为基于附图所示的方位或位置关系,仅是为了便于描述本发明和简化描述,而不是指示或暗示所指的装置或元件必须具有特定的方位、以特定的方位构造和操作,因此不能理解为对本发明的限制。此外,术语“第一”、“第二”仅用于描述目的,而不能理解为指示或暗示相对重要性。
在本发明的描述中,需要说明的是,除非另有明确的规定和限定,术语“安装”、“设置有”、“连接”等,应做广义理解,例如“连接”,可以是固定连接,也可以是可拆卸连接,或一体地连接;可以是机械连接,也可以是电连接;可以是直接相连,也可以通过中间媒介间接相连,可以是两个元件内部的连通。对于本领域的普通技术人员而言,可以具体情况理解上述术语在本发明中的具体含义。
实施例
一种核素标记的曲妥珠单抗,其为放射性核素131I标记的曲妥珠单抗。
一种核素标记的曲妥珠单抗的制备方法,包括以下步骤:
S1:称取Iodogen于试管中并用二氯甲烷溶解,取10μL的Iodogen加到反应管底部,用氮气吹干,使反应管底部形成均匀的薄膜,二氯甲烷的浓度为1g/L;
S2:向涂有一定量Iodogen的反应管中依次加入曲妥珠单抗10.0μg,Na131I溶液10μL,轻轻摇动,室温条件下反应8-10min,Na131I溶液的放射性活度为22.2MBq;
S3:反应完毕后,向S2的反应管中加入250μL PB液,静置,得到反应液,PB液的浓度为0.05mol/L,静置时间为5-8min;
S4:将S3所得的反应液用SephadexG-50柱进行纯化,得到目标产物,SephadexG-50柱使用前,先用0.01M的PBS溶液平衡柱体,然后再用0.01M的PBS液纯化出目标产物,平衡柱体的PBS溶液的量浓度为0.01M/L,pH为7.4;纯化目标产物的PBS溶液的量浓度为0.03M/L,pH为7.5;PBS溶液平衡柱体的具体方法为:每次加5mLPBS溶液,通过重力流速流干,重复3-5次。
当标记率大于92%,用SephadexG-50柱纯化后,使目标产物的放射性化学纯度大于98%
在本发明使用质子核素131I,131I可同时发射β和γ两种射线,β射线在组织中的射程仅为2-3mm,既保证了能在局部产生足够的电离辐射生物学效应,达到抑制及破坏病变组织的目的,又避免对邻近正常组织造成不必要的辐射,是目前较理想的RIT用放射性核素;131I—Herceptin在体外对高表达HER-2的人卵巢癌细胞SKOV3有明显的抑制作用,且131I-Herceptin能选择性的长时间浓聚于移植瘤部位;用131I标记Herceptin并作用于转移性胃癌和直肠癌,证实其具有良好的靶向作用,得到了很好的瘤体与正常组织间的吸收比率,实现靶向肿瘤药物的个体化、精准化治疗。
以上显示和描述了本发明的基本原理和主要特征和本发明的优点。本行业的技术人员应该了解,本发明不受上述实施例的限制,上述实施例和说明书中描述的只是说明本发明的原理,在不脱离本发明精神和范围的前提下,本发明还会有各种变化和改进,这些变化和改进都落入要求保护的本发明范围内。本发明要求保护范围由所附的权利要求书及其等效物界定。
Claims (9)
1.一种核素标记的曲妥珠单抗,其特征在于:其为放射性核素131I标记的曲妥珠单抗。
2.一种核素标记的曲妥珠单抗的制备方法,其特征在于:包括以下步骤:
S1:称取Iodogen于试管中并用二氯甲烷溶解,取10μL的Iodogen加到反应管底部,用氮气吹干,使反应管底部形成均匀的薄膜;
S2:向涂有一定量Iodogen的反应管中依次加入曲妥珠单抗10.0μg,Na131I溶液10μL,轻轻摇动,室温条件下反应8-10min;
S3:反应完毕后,向S2的反应管中加入250μL PB液,静置,得到反应液;
S4:将S3所得的反应液用SephadexG-50柱进行纯化,得到目标产物。
3.根据权利要求2所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:当标记率大于92%,用SephadexG-50柱纯化后,使目标产物的放射性化学纯度大于98%。
4.根据权利要求2所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:在所述S1中,所述二氯甲烷的浓度为1g/L。
5.根据权利要求2所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:在所述S2中,所述Na131I溶液的放射性活度为22.2MBq。
6.根据权利要求2所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:在所述S3中,所述PB液的浓度为0.05mol/L,静置时间为5-8min。
7.根据权利要求2所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:在所述S4中,所述SephadexG-50柱使用前,先用0.01M的PBS溶液平衡柱体,然后再用0.01M的PBS液纯化出目标产物。
8.根据权利要求7所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:所述平衡柱体的PBS溶液的量浓度为0.01M/L,pH为7.4;所述纯化目标产物的PBS溶液的量浓度为0.03M/L,pH为7.5。
9.根据权利要求8所述的一种核素标记的曲妥珠单抗的制备方法,其特征在于:所述PBS溶液平衡柱体的具体方法为:每次加5mLPBS溶液,通过重力流速流干,重复3-5次。
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