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Novel EGFR-MAPK kinase and ABC transporter inhibitors for HepG2 resistant to Erlotinib

Trevor Ostlund¹, Kakan SutraDhar¹, Sara M Elgazwi²

¹Department of Chemistry & Biochemistry, South Dakota State University, Brookings, South Dakota, USA.

Drug Development Research

|December 9, 2022

Abstract:

Hepatocellular carcinoma (HCC) is the third-leading cause of cancer death in the world, with outlook for most patients having a 5-year survivability of less than 5%. In a previous study from our laboratory, novel estrone inspired analogs act as epidermal growth factor receptor (EGFR) inhibitors in HepG2 cells. This study focuses on the effect of these analogs on an HCC cell line resistance to Erlotinib. Lead compounds MMA132 and MMA102 showed 13 and 20 µM IC₅₀ values, respectively against HepG2-R resistant to Erlotinib. These compounds showed cell cycle arrest of the G2 phase up to 54%, and inhibited cell migration of HepG2-R cells up to 48 h. Western blot analysis revealed that MMA132 reduced total EGFR content after 48 h, while MMA102 inhibited MEK kinase by 84% after 48 h. Western blot analysis also revealed that multidrug resistance protein 2 (MRP2) is overexpressed in HepG2-R, suggesting that ABC transporters play a likely cause in drug resistance. MMA102 showed significant inhibition of both P-glycoprotein (83%) and ABCG2 (53%), two additional ABC transporters. Additionally, MMA102 and MMA132 were used in a combination therapy with MK571(MRP1/2 inhibitor) and produced IC₅₀ values of 18 and 10 µM, respectively, better than either MMA102/132 or MK571 alone. To validate our findings, we conducted molecular dynamic simulations with MMA102 and MMA132 in MEK, P-glycoprotein, MRP1, and MRP2. Results coincided with biological findings in which MMA102 orientation is favored in both MEK and P-glycoprotein pockets, whereas MMA132 likely binds with MRP2, as likely suggested by the combinatorial study.

Insights

Novel estrone-inspired analogs, MMA102 and MMA132, show promise in overcoming Erlotinib resistance in hepatocellular carcinoma (HCC) by inhibiting key growth pathways and drug transporters.

Area of Science:

Oncology
Molecular Biology
Pharmacology

Background:

Hepatocellular carcinoma (HCC) presents a significant global health challenge with poor prognosis.
Erlotinib resistance is a major obstacle in HCC treatment.
Novel therapeutic strategies are needed to overcome drug resistance in HCC.

Purpose of the Study:

To evaluate the efficacy of novel estrone-inspired analogs (MMA102, MMA132) against Erlotinib-resistant HCC cells.
To investigate the molecular mechanisms underlying the action of these analogs, including their effects on EGFR, MEK, and ABC transporters.
To explore the potential of combination therapy with these analogs and known inhibitors.

Main Methods:

In vitro studies using Erlotinib-resistant HCC cell lines (HepG2-R).
Cell viability assays (IC50 determination), cell cycle analysis, and cell migration assays.
Western blot analysis to assess protein expression (EGFR, MEK, MRP2, P-glycoprotein, ABCG2).
Molecular dynamic simulations to predict binding interactions with target proteins.

Main Results:

MMA102 and MMA132 demonstrated significant anti-proliferative effects against HepG2-R cells with IC50 values of 20 µM and 13 µM, respectively.
Compounds induced G2 cell cycle arrest and inhibited cell migration.
MMA132 reduced total EGFR, while MMA102 inhibited MEK kinase.
Overexpression of MRP2 was observed in HepG2-R cells; MMA102 inhibited P-glycoprotein and ABCG2.
Combination therapy with MMA102/MMA132 and MK571 showed enhanced efficacy.

Conclusions:

Estrone-inspired analogs MMA102 and MMA132 are effective against Erlotinib-resistant HCC.
These compounds target key signaling pathways (EGFR, MEK) and multidrug resistance proteins (ABC transporters).
Combination therapy holds potential for improved HCC treatment outcomes.

Read full analysis

Keywords:

ABC transporters hepatocellular carcinoma molecular dynamics

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