文献：Nanotechnology-mediated immunochemotherapy with Ingenol-3-Mebutate for Systematic Anti-tumor Effects

文献链接：

https://www.sciencedirect.com/science/article/abs/pii/S016836591930255X

作者：Mian Yua, Miaoqing Zhao, Rilei Yua, Shaochen Chua, Jiahao Xua, Ming Xiad,Cheng Wang

相关产品：Cy7 labeled I3A

原文摘要：

 Cancer-Immunotherapy was the most exciting topic. However, either insensitivity due to singleness of therapeutic target or immune evasion leads to the failure of the treatment. Ingenol-3-mebutate (I3A) can inhibit cancer through synergy between immunotherapy and chemotherapy, however, the speculation and accurate mechanism haven't been confirmed in vivo limited by its hydrophobicity and pH-instability, which also hindered its clinical translation. Herein we developed a polymeric micelle with ‘acidic core’ provided by single alcoholic hydroxyl (-CH(CH3)-OH) encapsulating I3A (I3A-PM), which successfully overcome the aforementioned problems and reduce the toxicity in vivo. To test the synergy, S180 tumor-bearing mice were subjected to I3A-PM through intravenous and intratumoral administration, we found I3A-PM presented significant antitumor effect,and promoted Th1 polarization by upregulating the level of Th1 cytokines (IL-12, IL-2, IFN-γ and TNF-α), and accelerated the expansion of CD4+ and CD8+ T cells, meanwhile, I3A-PM depleted regulatory T cells, Th2 cytokine IL-6 through inhibiting TGF-β signaling pathway. Furthermore, we appealed to virtual screening of tumor target, and found a new pathway of I3A as a TGF-β receptor type I inhibitor to improve immunostimulatory effects. These results demonstrated I3A-PM as a promising nanoagent for cancer immunotherapy strategy. The synergistic therapeutic effects are encouraged to further evaluate in different cancer model compared with commercial products to facilitate research finding (I3A-PM) entering the clinic.  

Cy7 labeled I3A：Cy7 是一种荧光染料。Cy7 属于花菁染料（Cyanine Dyes）家族，它的最大吸收波长在 750 - 770nm 左右，发射波长在 780 - 800nm 左右，在近红外光区有很好的荧光性能。这种近红外荧光特性使得它在生物成像等领域很有优势，因为生物组织在近红外波段的自发荧光相对较弱，能减少背景干扰，提高检测的灵敏度和分辨率。I3A指一种化合物， 3 - 吲哚甲醛（indole - 3 - carbaldehyde）。“labeled” 表示标记的意思，即 I3A 这个分子被 Cy7 染料进行了标记。基于Cy7 labeled I3A的性能，介绍如下：

图：作用机制

Cy7 labeled I3A标记I3A-PM：

 I3A - PM 的预处理：如 I3A - PM 表面有杂质或者需要激活某些官能团才能进行标记反应，需要进行预处理。例如，如果 I3A - PM 是纳米颗粒，可能需要用缓冲溶液洗涤几次，去除表面的稳定剂或其他杂质。

标记反应：如果是直接标记 I3A - PM，反应步骤与 I3A 的标记类似。将 I3A - PM 分散在缓冲溶液中，加入 Cy7 染料溶液，控制合适的摩尔比、反应温度和时间进行标记反应。如果是将标记后的 I3A 与 I3A - PM 结合，需要考虑两者的结合方式。例如，如果是通过静电相互作用结合，要确保反应体系的 pH 和离子强度有利于静电吸附。可以将标记后的 I3A 溶液缓慢加入到 I3A - PM 分散液中，在温和搅拌或涡旋混合的条件下反应。

反应后处理：反应结束后，离心收集标记后的产物。用缓冲溶液洗涤几次，去除未反应的 Cy7 染料和 I3A。

图：电镜图像

结论：

该文献成功制出I3A-PM，Cy7 labeled I3A成功标记了I3A-PM，方便后续实验的进行。实验使用成像系统实时监测 I3A - PM 在体内的动态分布，观察富集情况，判断靶向作用的情况。结果表明，I3A-PM可作为一种有前途的cancer免疫Treatment 策略的纳米化合物。