Self-Assembling Derivative of Hydrocortisone as Glucocorticoid Receptor-Targeted Nanotherapeutics for Synergistic, Combination Therapy against Colorectal Tumor

Abstract

Hydrocortisone, a natural glucocorticoid secreted by adrenal and extra-adrenal tissues, locally governs the transcription of genes involved in inflammation, immune response, metabolism, and energy homeostasis via binding to its cognate glucocorticoid receptor (GR). In this study, we show that modified hydrocortisone (HC16), a cancer-selective cytotoxic molecule, showed synergism in combination with drugs like Doxorubicin and docetaxel, self-assembled into vesicles, entrapped docetaxel and complexed with anti-cancer plasmid DNA for enhanced killing of cancer cells. These vesicles exhibited GR-mediated nuclear localization, delivery of the p53 gene, and also inhibited cell viability selectively in RKO, HCT15, and CT26 colon cancer cells but not in normal cells like CHO and HEK293T. Apart from exerting its own anti-cancer activity, the self-assembled HC16 vesicles loaded with docetaxel sensitized the cancer cells to its drug cargo by downregulating the drug metabolizing CYP3A4 gene. This indirectly reduces the risk of nonspecific adverse effects in normal cells, as the viability of sensitized cancer cells could be significantly reduced even in low doses of cytotoxic docetaxel. The near infrared (NIR)-dye-associated self-assemblies accumulated in a colon tumor with higher orders of NIR intensity compared to those in a colon of healthy mice. Thereafter, the treatment of HC16− docetaxel−p53 vesicle/DNA complex led to significant tumor regression, which resulted in a cecum/body weight ratio in tumorbearing mice similar to that of healthy mice measured at 24 h postcompletion of treatment. There was an up to 2.5-fold enhancement in the overall survivability of colon-tumor-bearing mice treated with HC16−docetaxel−p53 vesicle/DNA complexes when compared against the pristine docetaxel-treated groups. Further, the HC16−docetaxel−p53 vesicle/DNA complex-treated group showed reduced nuclear accumulation of cell proliferation marker Ki67, reduced protein levels of prosurvival and mesenchymal proteins like Bcl-2, PARP, vimentin, and N-cadherin, and increased the levels of pro-apoptotic activated caspases as compared to the pristine docetaxel-treated groups. The therapeutic package described herein is expected to find future use as a rational, multifaceted, GR-targeted approach for inhibiting colon tumor progression.

Progesterone-Cationic Lipid Conjugate-Based Self-Aggregates for Cancer Cell-Selective Uptake through Macropinocytosis and the Antitumour Effect

“…While exploring the reason for the selective killing of cancer cells, a nanoaggregate-forming ability of these derivatives was noted. It was interesting because self-aggregating properties have been shown by us in cationic lipid conjugates of other small molecules and among natural steroids, hydrocortisone, before this study. − However, we are the first to report herein that another natural steroid hormone, progesterone, in its cationic lipid-conjugated derivatives, also exhibits similar self-aggregating properties. Presented herein are the details on the studies performed on the most potent derivative, i.e., PR10 nanoaggregates, and the mechanism involved in its cancer cell-selective uptake that led to its potent anticancer activity.…”

Targeting steroid hormone receptors for anti‐cancer therapy—A review on small molecules and nanotherapeutic approaches

“…A unique approach could thus be to access an alternate pathway by targeting GR, thus achieving efficient antitumor effect in CT26 colon tumor model using ESC8 and Dex-associated nanoformulation (Jinka et al, 2021). Same effect was also shown using hydrocortisone (a GR hormone)-conjugated cationic lipid based unique self-aggregated formulation (Sridharan et al, 2021). Tailored small molecules designed against custom mutant profiles if combined with such SHRtargeted therapeutic approaches might result in overall improved outcomes in terms of patient survival, even for cancers possessing gene mutations in future.…”

“…SAR study indicated HYC16, the aliphatic 16 carbon chain based cationic lipid-modified hydrocortisone, to be the most potent and selective in killing cancer cells and potently inhibiting angiogenesis (Rathore et al, 2016). As shown by other cationic lipid-modified GCs, for example, DX10, the same molecule HYC16 also exhibits potent drug sensitizing ability in cancer cells through reversal of EMT (Sridharan et al, 2021). Innovatively, HYC16 also exhibits a unique self-aggregating property, which is described in details later under Section 5.1.4, thus suggesting the advent of novel class of nanotherapeutics derived from steroid hormone.…”

“…The self-aggregate shows GR-targeted transfection in cancer cells, carries other hydrophobic drugs, can induce DX-alike drug sensitization through EMT-reversal and typical down regulation of drug metabolizing genes such as CYP3A4 and CYP1A1 selectively in cancer cells. Using anticancer p53 plasmid, and docetaxel as added anticancer drug, the holistic HYC16 formulation led to potent reduction in orthotopic colon tumor burden, and increased survivability of tumor-bearing mice, thereby once more proving the versatility of GR-targeted aggregates in the treatment of cancer (Sridharan et al, 2021). Hence, it is indeed worthwhile to check if the lipid modification strategy adopted to other SHRtargeted small molecules also produces novel SHR-targeted nanotherapeutics or not.…”

Development, optimization and validation of modified QuEChERS based UPLC-MS/MS for simultaneous determination of nine steroid hormones in milk powder and milk

“…10 HC is a synthetic and naturally occurring glucocorticoid, which has anti-inflammatory, immunosuppressive, anti-allergic, and anti-shock effects. 11 17α-Hydoxyprogesterone (OHP) is a derivative of progesterone transformed by 17α-hydroxylase. Overuse has caused the accumulation of steroid hormones in animals, which disturbs the human endocrine system via the food chain.…”