Synergistic inhibition of metastatic melanoma by carvacrol and chloroquine - an in vitro and in silico investigation of apoptosis and molecular targets

Sep 22, 2026

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Drug resistance is a major obstacle in treating metastatic melanoma, driving the need for new therapeutic strategies. This study evaluates the potential synergy between the plant-derived monoterpene carvacrol and the autophagy inhibitor chloroquine against metastatic melanoma. Cytotoxic and proapoptotic effects are tested in WM9 cells using WST-1 and Annexin V/7-AAD flow cytometry, quantified with a Q-value. In silico docking identifies shared high-affinity protein targets and proposes multi-target disruption of survival and signaling pathways.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages13

Reading Time33 min

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Abstract

Introduction

Drug resistance and need for new strategies

Carvacrol as a multi-target anticancer compound

Malignant melanoma epidemiology and clinical challenge

Medical Oncology (2026) 43:113 [https://doi.org/10.1007/s12032-025-03213-2](https://doi.org/10.1007/s12032-025-03213-2) ORIGINAL PAPER Synergistic inhibition of metastatic melanoma by carvacrol and chloroquine: an in vitro and in silico investigation of apoptosis and molecular targets Patrycja Kłos1 · Siarhei Dabravolski2 · Magdalena Perużyńska3 · Alicja Przekop1 · Paweł Kostka1 · Violetta Dziedziejko1 · Marek Droździk3 · Dariusz Chlubek1 · Krzysztof Safranow1 Received: 31 July 2025 / Accepted: 14 December 2025 / Published online: 13 January 2026 © The Author(s) 2026 Abstract The development of drug resistance remains a major obstacle in the treatment of metastatic melanoma, necessitating novel therapeutic strategies. Combining natural compounds with agents that disrupt cancer cell survival mechanisms, such as autophagy inhibitors, offers a promising approach. This study aimed to evaluate the potential synergy between the plantderived monoterpene carvacrol and the autophagy inhibitor chloroquine against metastatic melanoma. The cytotoxic and proapoptotic effects of carvacrol, chloroquine, and their combination were investigated in the WM9 metastatic melanoma cell line using WST-1 assays and Annexin V/7-AAD flow cytometry. The nature of the drug interaction was quantified using the Q-value. In silico molecular docking was performed to identify potential protein targets and elucidate the mechanisms underlying the observed effects. While carvacrol monotherapy exhibited weak cytotoxicity, its combination with non-toxic concentrations of chloroquine resulted in a potent and synergistic reduction in WM9 cell viability. This enhanced cytotoxicity was attributed to a significant, synergistic induction of apoptosis. In silico analysis predicted that both carvacrol and chloroquine bind with high affinity to common molecular targets, including Insulin-Like Growth Factor 1 Receptor and Sirtuin-2 . Chloroquine was also predicted to interact strongly with HSP90 . Our findings demonstrate that chloroquine potentiates the anticancer activity of carvacrol in metastatic melanoma cells. The synergy observed is likely mediated by a multi-targeted mechanism involving the simultaneous disruption of key cancer survival and signaling pathways. This study establishes the carvacrol-chloroquine combination as a novel and effective strategy that warrants further preclinical investigation for the treatment of resistant melanoma. Keywords Melanoma · Carvacrol · Chloroquine · Synergism · Apoptosis 􀀍 Patrycja Kłos [patrycja.klos@pum.edu.pl](patrycja.klos@pum.edu.pl) Siarhei Dabravolski [sergedobrowolski@gmail.com](sergedobrowolski@gmail.com)[ ](sergedobrowolski@gmail.com)Magdalena Perużyńska [magdalena.peruzynska@pum.edu.pl](magdalena.peruzynska@pum.edu.pl)[ ](magdalena.peruzynska@pum.edu.pl)Alicja Przekop[alaperskawiec@gmail.com](alaperskawiec@gmail.com) Paweł Kostka [pawel.kostka99@gmail.com](pawel.kostka99@gmail.com)[ ](pawel.kostka99@gmail.com)Violetta Dziedziejko [violetta.dziedziejko@pum.edu.pl](violetta.dziedziejko@pum.edu.pl)[ ](violetta.dziedziejko@pum.edu.pl)Marek Droździk [marek.drozdzik@pum.edu.pl](marek.drozdzik@pum.edu.pl) Dariusz Chlubek [dariusz.chlubek@pum.edu.pl](dariusz.chlubek@pum.edu.pl) Krzysztof Safranow [krzysztof.safranow@pum.edu.pl](krzysztof.safranow@pum.edu.pl) 1 Department of Biochemistry and Medical Chemistry, Pomeranian Medical University in Szczecin, Powstańców Wlkp. 72, Szczecin 70-111, Poland 2 Department of Biotechnology Engineering, Braude Academic College of Engineering, Snunit 51, P.O. Box 78, Karmiel 2161002, Israel 3 Department of Clinical and Experimental Pharmacology, Pomeranian Medical University in Szczecin, Powstańców Wlkp. 72, Szczecin 70-111, Poland Introduction Despite significant advances in synthetic drug development, plant-derived essential oils remain a focus of intense research as a rich source of potential therapeutic agents. These natural compounds exhibit a broa

FAQs

What is the main goal of combining carvacrol and chloroquine in this study?

To assess whether carvacrol and chloroquine act synergistically against metastatic melanoma by targeting mechanisms related to cancer cell survival, including autophagy.

How were cytotoxicity and apoptosis measured experimentally?

Cytotoxic effects were evaluated using WST-1 assays, while apoptosis-related changes were measured with Annexin V/7-AAD flow cytometry in the WM9 metastatic melanoma cell line.

What role does the in silico docking analysis play?

It predicts protein targets and binding affinities for carvacrol and chloroquine, helping explain mechanisms behind the observed synergy, including interactions with targets such as IGF1R, Sirtuin-2, and HSP90.

Sep 22, 2026

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