title: Cathepsin D inhibitors as potential therapeutics for breast cancer treatment: Molecular docking and bioevaluation against triple-negative and triple-positive breast cancers.
abstract: The main aim of this study was to discover small molecule inhibitors against Cathepsin D (CatD) (EC.3.4.23.5), a clinically proven prognostic marker for breast cancer, and to explore the mechanisms by which CatD could be a useful therapeutic target for triple-positive and triple-negative breast cancers (TPBC & TNBC). The crystal structure of CatD at 2.5 Å resolution (PDB: 1LYB), which was complexed with Pepstatin A, was selected for computer-aided molecular modeling. The methods used in our study were pharmacophore modeling and molecular docking. Virtual screening was performed to identify small molecules from an in-house database and a large commercial chemical library. Cytotoxicity studies were performed on human normal cell line HEK293T and growth inhibition studies on breast adenocarcinoma cell lines, namely MCF-7, MDA-MB-231, SK-BR-3, and MDA-MB-468. Furthermore, RT-PCR analysis, in vitro enzyme assay, and cell cycle analysis ascertained the validity of the selected molecules. A set of 28 molecules was subjected to an in vitro fluorescence-based inhibitory activity assay, and among them six molecules exhibited >50 % inhibition at 25μM. These molecules also exhibited good growth inhibition against TPBC and TNBC cancer types. Among them, molecules 1 and 17 showed single-digit micromolar GI50 values against MCF-7 and MDA-MB-231 cell lines.
authors.affiliation: Computer-Aided Drug Design Laboratory, Department of Pharmacy, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, R.R. District, Hyderabad, 500078, Telangana, India.
authors.forename: Hasitha Shilpa
authors.initials: HS
authors.lastname: Anantaraju
authors.affiliation: Computer-Aided Drug Design Laboratory, Department of Pharmacy, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, R.R. District, Hyderabad, 500078, Telangana, India.
authors.forename: Madhu Babu
authors.initials: MB
authors.lastname: Battu
authors.affiliation: Incozen Therapeutics (P)Ltd, Shameerpet, R.R. District, Hyderabad, 500078, Telangana, India.
authors.forename: Srikant
authors.initials: S
authors.lastname: Viswanadha
authors.affiliation: Computer-Aided Drug Design Laboratory, Department of Pharmacy, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, R.R. District, Hyderabad, 500078, Telangana, India.|Yogee'S Bioinnovations Private Limited, Room No. 5, Technology Business Incubator, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, Hyderabad, 500078, Telangana, India.
authors.forename: Dharmarajan
authors.initials: D
authors.lastname: Sriram
authors.affiliation: Computer-Aided Drug Design Laboratory, Department of Pharmacy, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, R.R. District, Hyderabad, 500078, Telangana, India. pyogee@hyderabad.bits-pilani.ac.in.|Yogee'S Bioinnovations Private Limited, Room No. 5, Technology Business Incubator, Birla Institute of Technology & Science- Pilani, Hyderabad Campus, Shameerpet, Hyderabad, 500078, Telangana, India. pyogee@hyderabad.bits-pilani.ac.in.
authors.forename: Perumal
authors.initials: P
authors.lastname: Yogeeswari
mesh_terms: D002402:Cathepsin D / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004354:Drug Screening Assays, Antitumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D062105:Molecular Docking Simulation; D011480:Protease Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D011487:Protein Conformation; D064726:Triple Negative Breast Neoplasms / Q000473:pathology*; D059016:Tumor Microenvironment / Q000187:drug effects
chemical_list: D011480:Protease Inhibitors; D002402:Cathepsin D
keywords: ASINEX; Breast cancer; CatD; Cathepsin D; Docking; Pharmacophore model