Comprehensive Physicochemical Characterization, In Vitro Membrane Permeation, and In Vitro Human Skin Cell Culture of a Novel TOPK Inhibitor, HI-TOPK-032.
Comprehensive Physicochemical Characterization, In Vitro Membrane Permeation, and In Vitro Human Skin Cell Culture of a Novel TOPK Inhibitor, HI-TOPK-032.
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DOI：
10.3390/ijms242115515
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发表时间：
2023-10-24
影响因子：
5.6
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中科院分区：
文献类型：
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非黑色素瘤皮肤癌(NMSC)是最常见的皮肤癌，在美国每年约有540万人被诊断为皮肤癌。新近开发的T淋巴因子激活的杀伤细胞起源蛋白激酶(TOPK)抑制剂HI-TOPK-032可以有效地抑制结肠癌细胞生长，诱导结肠癌细胞和紫外线诱导的鳞状细胞癌(SCC)的凋亡。本研究旨在研究HI-TOPK-032的理化性质、渗透行为和细胞毒性，为开发适合的皮肤靶向给药制剂奠定基础。采用扫描电子显微镜(SEM)、能量色散X射线(EDX)、差示扫描量热仪(DSC)、热台显微镜(HSM)、X射线粉末衍射仪(XRPD)、卡尔-费雪(KF)库仑滴定、拉曼光谱、共聚焦拉曼显微镜(CRM)、衰减全反射傅里叶变换红外光谱(ATR-FTIR)、傅里叶变换红外光谱(FTIR)等手段对HI-TOPK-032进行了表征。用人皮肤角质形成细胞系(HaCaT)和原代正常人表皮角质形成细胞(NHEKs)进行2D细胞培养，评价HI-TOPK-032对体外细胞活力的剂量效应。在HaCAT人皮肤细胞系上测量了气液界面跨皮电阻(TEER)随剂量和时间的变化。用Strat-M®仿生膜和Franz细胞体外扩散系统测试了HI-TOPK-032的渗透行为。理化评价结果证实了药物的无定形性质和样品的均一性，具有所有特征化学峰。体外细胞存活率测定结果证实，HI-TOPK-032在10微米以下的细胞存活率为100%。在此基础上，建立了一种快速、特异、准确、有效的反相高效液相色谱定量测定HI-TOPK-032的方法。这是HI-TOPK-032的首次系统和全面的表征以及这些发现的报告。
Nonmelanoma skin cancers (NMSC) are the most common skin cancers, and about 5.4 million people are diagnosed each year in the United States. A newly developed T-lymphokine-activated killer cell-originated protein kinase (TOPK) inhibitor, HI-TOPK-032, is effective in suppressing colon cancer cell growth, inducing the apoptosis of colon cancer cells and ultraviolet (UV) light-induced squamous cell carcinoma (SCC). This study aimed to investigate the physicochemical properties, permeation behavior, and cytotoxicity potential of HI-TOPK-032 prior to the development of a suitable topical formulation for targeted skin drug delivery. Techniques such as scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) spectroscopy, differential scanning calorimetry (DSC), hot-stage microscopy (HSM), X-ray powder diffraction (XRPD), Karl Fisher (KF) coulometric titration, Raman spectrometry, confocal Raman microscopy (CRM), attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), and Fourier transform infrared microscopy were used to characterize HI-TOPK-032. The dose effect of HI-TOPK-032 on in vitro cell viability was evaluated using a 2D cell culture of the human skin keratinocyte cell line (HaCaT) and primary normal human epidermal keratinocytes (NHEKs). Transepithelial electrical resistance (TEER) at the air–liquid interface as a function of dose and time was measured on the HaCAT human skin cell line. The membrane permeation behavior of HI-TOPK-032 was tested using the Strat-M® synthetic biomimetic membrane with an in vitro Franz cell diffusion system. The physicochemical evaluation results confirmed the amorphous nature of the drug and the homogeneity of the sample with all characteristic chemical peaks. The in vitro cell viability assay results confirmed 100% cell viability up to 10 µM of HI-TOPK-032. Further, a rapid, specific, precise, and validated reverse phase-high performance liquid chromatography (RP-HPLC) method for the quantitative estimation of HI-TOPK-032 was developed. This is the first systematic and comprehensive characterization of HI-TOPK-032 and a report of these findings.
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影响因子：
4.6
作者：
Colombo I;Sangiovanni E;Maggio R;Mattozzi C;Zava S;Corbett Y;Fumagalli M;Carlino C;Corsetto PA;Scaccabarozzi D;Calvieri S;Gismondi A;Taramelli D;Dell'Agli M
通讯作者：
Dell'Agli M
影响因子：
11.2
作者：
Kim, Dong Joon;Li, Yan;Dong, Zigang
通讯作者：
Dong, Zigang
影响因子：
3.8
作者：
Côté, S;Simard, C;Lemieux, R
通讯作者：
Lemieux, R
影响因子：
8
作者：
Meenach SA;Vogt FG;Anderson KW;Hilt JZ;McGarry RC;Mansour HM
通讯作者：
Mansour HM
影响因子：
4.8
作者：
Abe, Y;Matsumoto, S;Ueda, N
通讯作者：
Ueda, N