Abstract NTOC-077: ORALLY ACTIVE LOW MOLECULAR WEIGHT HEPARIN TAUROCHOLATE AND DEOXYCHOLATE CONJUGATE FOR THE INHIBITION OF THE PROGRESSION OF METASTATIC OVARIAN CANCER
Authors: Alam Farzana , Haque Mohammad Rezwanul , Al-Hilal Taslim A. , Ahsan Fakhrul , Byun Youngro 
Published: June 2017

PURPOSE: Ovarian cancer is one of the major aggressive cancer which is not manageable blocking a single pathway. Recently, chemically modified low molecular weight heparin (LMWH) derivatives have shown good anti-cancer activity targeting various biochemical parameters, such as, transforming growth factor beta 1 (TGFβ-1), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), heparin-like growth factor (HGF), CXCL-12, and CXCL-9. In this study, we have focused on TGFβ-1 and VEGF that are known to promote metastatic ovarian cancer progression and use an orally LMWH-taurocholate conjugated with tetrameric deoxycholic acid, namely LHTD4, to successfully inhibit the progression at different stages of ovarian metastasis cancer.

EXPERIMENTAL PROCEDURES: We performed the binding affinity of TGFβ-1 and VEGF with LHTD4. We carried out in vitro phosphorylation assays of the consecutive receptors of TGFβ-1 and VEGF (TGF-β-1R1 and VEGFR2, respectively). Effects of LHTD4 on in vitro SKOV3-LUC cell migration (induced by TGFβ-1 and VEGF), spheroid formation, and the epithelial to mesenchymal transition (EMT) were investigated. We prepared a formulation of LHTD4 and a synthetic bile acid-based absorption enhancer, DCK (L-lysine- deoxycholylethylamine), to enable the oral absorption. The in vivo anti-metastatic effect of LHTD4 was evaluated in an intraperitoneal adhesion model and an orthotopic SKOV3-LUC ovarian cancer model. We measured the luciferase activity of the mice every week to check the progression of the cancer.

RESULTS: LHTD4 inhibited the migration of SKOV3-LUC cells 10-fold greater than the control group. LHTD4 also blocked the VEGF induced SKOV3-LUC spheroid formation. LHTD4 effectively inhibited ligand-induced VEGFR2 and TGFβ-1R1 phosphorylated. The expressions of EMT marker proteins such as Vimentin and Snail were prevented by LTHD4 treatment in in vitro studies with TGFβ-1 treated SKOV3-LUC cells. In orthotopic model ovarian metastasis model, the treatment with LHTD4 (5 mg/kg daily, orally) significantly inhibited the growth and organ metastasis progression compared to the control: LHTD4-treated mice showed 20-times lower luciferase activity than that of control mice. Finally, the intraperitoneal adhesion of the SKOV3-LUC cells was also decreased robustly in mice which received LHTD4 daily.

CONCLUSION: LHTD4 is one of the promising candidate for the inhibition of the progression of epithelial ovarian cancer. We report that the oral formulation of LHTD4/DCK complex could be a good choice for the treatment of metastatic ovarian cancer as it can target multiple pathways of metastasis. LHTD4 primarily showed anti-metastatic effects via blocking of TGFβ-1 and VEGF signaling pathway at different stages of metastasis progress.

Citation Format: Farzana Alam, Mohammad Rezwanul Haque, Taslim A. Al-Hilal, Fakhrul Ahsan, Youngro Byun. ORALLY ACTIVE LOW MOLECULAR WEIGHT HEPARIN TAUROCHOLATE AND DEOXYCHOLATE CONJUGATE FOR THE INHIBITION OF THE PROGRESSION OF METASTATIC OVARIAN CANCER [abstract]. In: Proceedings of the 11th Biennial Ovarian Cancer Research Symposium; Sep 12-13, 2016; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(11 Suppl):Abstract nr NTOC-077.