利用高光谱成像、分光光度法和分光荧光测定法研究白蛋白溶解的疏水性近红外吸收二拉德铂(II)复合物的细胞吸收。

IF 1.8 4区 化学 Q3 CHEMISTRY, ANALYTICAL Analytical Sciences Pub Date : 2024-06-19 DOI:10.1007/s44211-024-00621-8
Ryota Sawamura, Atsuko Masuya-Suzuki, Nobuhiko Iki
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引用次数: 0

摘要

由于具有生物可渗透性和微创性,波长在 700-1100 纳米之间的近红外(NIR)光在癌症诊断和治疗中备受关注。我们的研究小组以前曾报告过,疏水性二元铂(II)复合物 PtL2 是一种很有希望用于癌症光热疗法的药剂(L = 3,5-二溴-1,2-二氨基苯并喹喔啉酸自由基)。由于 PtL2 不发荧光,因此无法用荧光显微镜观察到细胞间对 PtL2 的吸收。在本研究中,我们利用高光谱成像技术对图像进行了分光光度分析,从而弄清了被牛血清白蛋白(BSA)溶解的 PtL2 的吸收和细胞内行为。所获图像中的光谱变化表明,在较长的培养期(> 4 小时)内,PtL2 的内化随之而来的是复合物的结晶化。此外,根据 PtL2 与 BSA 结合时的荧光淬灭分析,可以估算出结合常数 Kb = 5.91 × 104 M-1;Kb 比白蛋白类常见药物的 Kb 小两个数量级。考虑到 PtL2 的 Kb 较小且在水中的溶解度较低,我们最终提出了 PtL2 在细胞中的内化路径和归宿:PtL2 在细胞膜附近从 BSA 中释放,然后通过膜渗透被细胞吸收,继而饱和,导致结晶。
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Study on cellular uptake of a hydrophobic near-infrared-absorbing diradical-platinum(II) complex solubilized by albumin using hyperspectral imaging, spectrophotometry, and spectrofluorimetry

Owing to its biopenetrability and minimal invasiveness, near-infrared (NIR) light in the region between 700–1100 nm has attracted attention in cancer diagnosis and therapy. Our group previously reported that the hydrophobic diradical-platinum(II) complex PtL2 is a promising agent for cancer photothermal therapy (L = 3,5-dibromo-1,2-diiminobenzosemiquinonate radical). Because PtL2 does not fluoresce, its intercellular uptake of PtL2 cannot be observed with a fluorescence microscope. In this study, we clarified the uptake and intracellular behavior of PtL2 solubilized by bovine serum albumin (BSA) using hyperspectral imaging enabling spectrophotometric analysis of the image. The spectral changes in the obtained images indicated that the internalization of PtL2 was followed by crystallization of the complex during the long incubation period (> 4 h). Additionally, the binding constant Kb = 5.91 × 104 M–1 could be estimated upon fluorescence quenching analysis of BSA upon binding of PtL2; Kb is two orders of magnitude smaller than that of albumin–common drugs. Considering the small Kb and low solubility of PtL2 in water, we ultimately proposed the internalization path and fate of PtL2 in the cell: release of PtL2 from BSA near cellular membranes and subsequent cellular uptake via membrane permeation followed by saturation, resulting in crystallization.

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来源期刊
Analytical Sciences
Analytical Sciences 化学-分析化学
CiteScore
2.90
自引率
18.80%
发文量
232
审稿时长
1 months
期刊介绍: Analytical Sciences is an international journal published monthly by The Japan Society for Analytical Chemistry. The journal publishes papers on all aspects of the theory and practice of analytical sciences, including fundamental and applied, inorganic and organic, wet chemical and instrumental methods. This publication is supported in part by the Grant-in-Aid for Publication of Scientific Research Result of the Japanese Ministry of Education, Culture, Sports, Science and Technology.
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