Theaflavin-3-gallate, a natural antagonist for Hsp90: In-silico and in-vitro approach
Graphical abstract
Introduction
Breast cancer metastasis is a terminal event and the leading cause of mortality in cancer patients; however, the mechanism is poorly understood [1]. In spite of progress in diagnosis and therapeutic modalities using newer technologies metastasis continue to be the major cause of cancer-related death until today [2]. Endocrine therapy, surgery, and chemotherapy are the main treatment approaches for breast cancer, and breast cancer that is identified early on always has an improved prognosis after surgery [3]. Owing to a lack of ample evidence on the mechanism that triggers the metastatic process in breast cancer, partial achievement has been made in the prevention and inhibition of metastatic events using target-specific inhibitors. Heat Shock Proteins (HSP) are among key targets of interest to many researchers who are putting effort to develop specific antagonists due to their potential role in tumorigenesis and metastasis [4].
Mammalian HSPs have been classified mainly in four families according to their molecular weight “Hsp90, Hsp70, Hsp60, and small HSP, Hsp90, a family of an important chaperone, were first identified in a complex form with steroid hormone receptors and oncoprotein viral Src kinase and it has been shown that HSPs were significantly associated with cancers [5,6]. In detail, heat shock protein (HSP) 90 has been found to play critical roles in protein folding, intracellular disposition, and proteolytic turnover of key regulators of cell growth and survival. Furthermore, many studies revealed that Hsp90 can regulate the conformation, stability, and function of various oncogenic proteins to participate in the proliferation, apoptosis, and metastasis of tumor cells [7]. Moreover, breast cancer is a sign that Hsp90 inhibitors should be discovered for a diversity of reasons such as inhibition of Hsp90 has also been known to modulate angiogenesis of breast cancer xenografts [8]. Beside, recent study also suggested that in breast cancer patient mRNA level of HSP90 was significantly upregulated in primary as well as bone metastases [9]. Since Hsp90 was recognized as a potential cancer target, numerous inhibitors targeting cytoplasmic Hsp90 have been established as anticancer agents but, none have shown satisfactory effectiveness to be approved by the FDA [10,11]. Therefore, it is the need of the hour to identify a potent lead compound as an hsp90 antagonist and evaluate its efficiency at the molecular & cellular levels. Interlinking biological experiments with computational modeling and investigation may lead to a decrease in numeral in vitro experiments, enhanced validation evaluations, and improved sensitivity.
Thus, this study aims to screen potential lead compounds targeting the Hsp90 protein using molecular docking and molecular dynamics simulation approach. 95 natural compounds were screened as potential lead compounds against the Hsp90 protein through various parameters such as binding energy and pattern. The therapeutic potential of the possible lead compound was further analyzed using an MTT assay, apoptotic, migration, colony formation, and cell cycle assay. The potent lead compound against Hsp90 may come up as a novel anti-metastatic agent in the future for breast cancer.
Section snippets
Retrieval of protein structures and its preparations
The Hsp90 protein target in complex with 4-(1,3-dihydro-2h-isoindol-2-ylcarbonyl)-6-(1-methylethyl) benzene-1,3-diol (PDB entry: 2xab, chain A) was retrieved from Protein Data Bank (PDB) [12]. The retrieved structures were pre-processed using Protein Preparation Wizard of Schrödinger suite (academic license) (Schrödinger Suite 2014). Furthermore, restrained minimization was carried out to obtain a geometrically-stable protein conformation. A curated ligand library consisting of 95 natural
Molecular docking of Hsp90 target
We performed molecular docking of Hsp90 target using a curated collection of 95 natural molecules with proven anti-cancer activity. Extensive docking calculations were carried out using the XP module of Schrödinger suite (academic license) with thorough examination of optimal intermolecular contacts with the target. The binding site of 2,4-dihydroxy benzamide molecule (co-crystal ligand of PDB entry: 2xab) was defined as the docking site which encompassed all amino acids with 8 Å distance to
Discussion
The heat shock protein (Hsp90) belongs to a family of molecular chaperones, involved in regulating biological and functional cell processes such as cell growth, invasion, migration, and recurrence that are responsible for the progression of different malignancies [[31], [32]]. Numerous studies have emphasized the importance of Hsp90 in the last two decades; however, there was a great difference in these reports, as all these studies fail to take into consideration that Hsp90 is a group of
Author contribution
Conceptualization, Methodology, Validation, Writing - Original Draft, Software, Writing - Review & Editing, Data analysis, Software, Software, Visualization, Supervision, Investigation, Funding, Review & Editing.
Author statement
All persons who meet authorship criteria are listed as authors, and all authors certify that they have participated sufficiently in the work to take public responsibility for the content, including participation in the concept, design, analysis, writing, or revision of the manuscript. Furthermore, each author certifies that this material or similar material has not been and will not be submitted to or published in any other publication before its appearance in the chemico-biological interaction.
Declaration of competing interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgement
Kinjal Bhadresha is thankful to Indian Council of Medical Research (ICMR; New Delhi) for the financial assistance in the form of Senior Research fellowship (ortho/2018/NCD-I). The authors thank the Department of Botany, Bioinformatics and Climate Change Impacts Management, Gujarat University for providing the supercomputing facility support to carry out this study.
References (56)
Cancer metastases: challenges and opportunities
Acta Pharm. Sin. B
(2015)- et al.
Heat shock proteins in cancer: chaperones of tumorigenesis
Trends Biochem. Sci.
(2006) - et al.
A predictive biomarker panel for bone metastases: liquid biopsy approach
J. Bone Oncol.
(2021) - et al.
A simple technique for quantitation of low levels of DNA damage in individual cells
Exp. Cell Res.
(1988) - et al.
Design strategies to target crystallographic waters applied to the Hsp90 molecular chaperone
Bioorg. Med. Chem. Lett
(2011) - et al.
HSP90 and HSP70 proteins are essential for stabilization and activation of WASF3 metastasis-promoting protein
J. Biol. Chem.
(2012) - et al.
Theaflavins induced apoptosis of LNCAP cells is mediated through induction of p53, down-regulation of NF-kappa B and mitogen-activated protein kinases pathways
Life Sci.
(2007) - et al.
Involvement of cell surface HSP90 in cell migration reveals a novel role in the developing nervous system
J. Biol. Chem.
(2004) - et al.
EGCG inhibited bladder cancer SW780 cell proliferation and migration both in vitro and in vivo via down-regulation of NF-κB and MMP-9
J. Nutr. Biochem.
(2017) - et al.
Epithelial-mesenchymal transition in breast cancer progression and metastasis
Chin. J. Cancer
(2011)
Prognostic factors in metastatic breast cancer: a prospective single-centre cohort study in a Finnish University Hospital
BMJ Open
Heat Shock Proteins as Novel Therapeutic Targets in Cancer
In Vivo
High expression of heat shock protein 90 is associated with tumor aggressiveness and poor prognosis in patients with advanced gastric cancer
PLoS One
HSP90 promotes cell glycolysis, proliferation and inhibits apoptosis by regulating PKM2 abundance via Thr-328 phosphorylation in hepatocellular carcinoma
Mol. Cancer
NVP-AUY922: a novel heat shock protein 90 inhibitor active against xenograft tumor growth, angiogenesis, and metastasis
Cancer Res.
Recent Advances in the Discovery of Novel HSP90 Inhibitors: an Update from. Curr
Discovery of (2, 4-Dihydroxy-5-isopropylphenyl)-[5-(4-methylpiperazin-1-ylmethyl)-1,3 dihydroisoindol-2-yl] methanone (AT13387), a novel inhibitor of the molecular chaperone hsp90 by fragment based drug design
J. Med. Chem.
Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments
J. Comput. Aided Mol. Des.
PubChem: a public information system for analyzing bioactivities of small molecules
Nucleic Acids Res.
Integrated modeling program, applied chemical theory (IMPACT)
J. Comput. Chem.
Nosé–Hoover chains: the canonical ensemble via continuous dynamics
J. Chem. Phys.
Constant pressure molecular dynamics algorithms
J. Chem. Phys.
Constant pressure molecular dynamics algorithms
J. Chem. Phys.
A multiple time step molecular dynamics algorithm for macromolecules
J. Phys. Chem.
Identification of novel aldose reductase inhibitors from spices: a molecular docking and simulation study
PLoS One
CellExpress: a Comprehensive Microarray-Based Cancer Cell Line and Clinical Sample Gene Expression Analysis Online System, Database
Metamizole (dipyrone) – cytotoxic and antiproliferative effects on HeLa, HT-29 and MCF-7 cancer cell lines
Biotechnol. Biotechnol. Equip.
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