Near-IR Photorelease Chemistry: Discovery and Applications

近红外光释放化学：发现和应用

• 批准号: 8938197
• 负责人: Martin Schnermann
• 金额: $ 84.4万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8938197
• 关键词: Address; Amines; Antibodies; Area; Biological; Biology; Cell Line; Cell Survival; Cellular biology; Chemicals; Chemistry; Clinical; Collaborations; Complex; Control Groups; Development; Drug Delivery Systems; Estrogen Receptors; Fluorescence; Gene Expression; Gene Expression Regulation; Genetic Recombination; Goals; Hydrolysis; Kinetics; Lead; Ligands; Light; Literature; Mass Spectrum Analysis; Mediating; Molecular Target; Operative Surgical Procedures; Penetration; Pharmaceutical Preparations; Phenols; Photobleaching; Photons; Process; Reaction; Reporter; Science; Staging; System; Techniques; Therapeutic; Tissues; Toxic effect; Ultraviolet Rays; base; cancer therapy; chemical reaction; cyanine; cytotoxic; fluorophore; irradiation; novel strategies; scaffold; small molecule; success

Photoremovable caging groups find extensive use in many fields ranging from cell biology to materials science. The general requirement of UV or blue light is a significant limitation due to associated toxicity and poor tissue penetration. By contrast, light between 650 and 900 nm, often referred to as the near-IR window, is cytocompatible and has significant tissue penetration (centimeters). Useful single photon reactions in the near-IR range would allow photorelease or "uncaging" approaches to be applied in complex biological settings. My lab is approaching this task by using chemical reactions involved in the photodecomposition or "photobleaching" of near-IR fluorophores. These oft-encountered light-initiated processes can occur with rapid kinetics, often undesirably so, and I believe represent an untapped opportunity for chemical biology. Our current efforts in this area are split in three aims. Aim 1: Development and mechanistic studies of near-IR photorelease chemistry. Over the past year, we have developed an uncaging reaction sequence initiated by 690 nm light using readily synthesized C4'-dialkylamine-substituted heptamethine cyanines. We have shown that a variety of phenol- and amine- containing small molecules are quickly uncaged upon irradiation with low energy light. Detailed mechanistic studies involving mass spectrometry, NMR, and absorbance techniques have shown that release occurs through regioselective C-C cleavage and then hydrolysis of the C4'-amine. While the photooxidative cleavage reaction had been previously described in cyanine photobleaching literature, these efforts are the first to use it for productive application. We are currently broadening the scope of the release process and examining aspects of the mechanism in detail using computational (collaboration with Dr. Joseph Ivanic) and experimental techniques. Aim 2: Application of uncaging reactions for targeted drug delivery. Light is used in a variety of cancer treatments ranging from established therapeutic techniques, such as PDT, to emerging areas such as fluorescence-guided surgery. Separately, the last decade has seen the clinical success of a number of antibody-drug conjugates. An enduring challenge is the development of precisely controlled linker strategies. To address this, we are applying our light-cleavable chemistry for targeted drug delivery (collaboration with Dr. Hisataka Kobayashi, Dr. Peter Choyke). This approach would merge the unique potency of small molecule drugs with the high spatial control afforded by light release and molecular targeting. The use of tissue penetrant, cytocompatible near-IR light is critical because existing uncaging chemistries using UV or blue light would not be suitable for this application. Towards this goal, we have shown that cell viability can be inhibited through light-dependent release of several cytotoxic small molecules. Aim 3: Near-IR light control of gene expression. Among many strategies to achieve precise regulation of gene expression, several studies over the past 15 years have used UV light-mediated uncaging of small molecules in combination with inducible gene expression systems. It is quite likely that near-IR uncaging will prove beneficial as these techniques progress in complex biological settings and organismal contexts. We have shown that our approach can be used to regulate gene expression through uncaging of an estrogen-receptor antagonist in a ligand-dependent CreERT/LoxP-reporter cell line (collaboration with Dr. Susan Mackem). These results set the stage for further applications that combine near-IR uncaging with widely available CreERT-mediated recombination approaches.

可光熟悉的笼子群在从细胞生物学到材料科学的许多领域中找到了广泛的使用。紫外线或蓝光的一般需求是由于毒性和组织渗透不良而受到的重大限制。相比之下，通常称为近红外窗口的650至900 nm之间的光是细胞相容的，具有明显的组织穿透力（厘米）。有用的近IR范围内的单个光子反应将允许在复杂的生物学环境中应用光线释放或“分离”方法。我的实验室正在通过使用涉及光电分解或近红外荧光团“光漂白”的化学反应来完成这项任务。这些经常遇到的光启动过程可能会以快速的动力学发生，通常是不必要的，我相信这是化学生物学的未开发的机会。我们目前在这一领域的努力分为三个目标。 AIM 1：近IR光释放化学的发展和机械研究。在过去的一年中，我们使用了容易合成的C4'Dialkylamine添加的己氨酸氰氨酸，开发了一个由690 nm光引发的分离反应序列。我们已经表明，在低能量光线下照射时，各种含有苯酚和胺 - 含有小分子很快就会液化。涉及质谱，NMR和吸光度技术的详细机械研究表明，释放是通过区域选择性的C-C裂解进行的，然后进行C4'-胺的水解。虽然先前在氰胺光漂白文献中描述了光氧化性裂解反应，但这些努力是第一个将其用于生产性应用的努力。目前，我们正在扩大发布过程的范围，并使用计算（与Joseph Ivanic博士的协作）和实验技术详细研究该机制的各个方面。 AIM 2：应用分解反应用于靶向药物。光用于各种癌症治疗，从已建立的治疗技术（例如PDT）到新兴区域，例如荧光引导手术。另外，在过去的十年中，许多抗体 - 药物结合物都取得了成功。持久的挑战是开发精确控制的接头策略。为了解决这个问题，我们正在将可转移的化学物质应用于靶向药物（与Hisataka Kobayashi博士，Peter Choyke博士的合作）。这种方法将将小分子药物的独特效力与光释放和分子靶向提供的高空间控制。使用组织渗透剂，细胞相容的近红外光非常关键，因为现有的使用UV或蓝光的化学效果不适合此应用。为了实现这一目标，我们已经表明，可以通过几个细胞毒性小分子的光依赖性释放来抑制细胞活力。 AIM 3：基因表达的近IR光控制。在实现基因表达的精确调节的许多策略中，过去15年中的几项研究都使用了紫外线介导的小分子与诱导基因表达系统结合使用。随着这些技术在复杂的生物环境和有机环境中的发展，近红外的未约会很可能会证明是有益的。我们已经表明，我们的方法可用于通过在配体依赖性的Creert/Loxp-Reporter细胞系中（与Susan Mackem博士的协作）中的雌激素受体拮抗剂进行脱离来调节基因表达。这些结果为进一步的应用奠定了阶段，这些应用结合了近红外未代与广泛可用的Creert介导的重组方法。