title: Novel di-tertiary-butyl phenylhydrazones as dual cyclooxygenase-2/5-lipoxygenase inhibitors: synthesis, COX/LOX inhibition, molecular modeling, and insights into their cytotoxicities.
abstract: Although dual inhibition of Cyclooxygenase-2 (COX-2) and 5-Lipoxygenase (5-LOX) enzymes is highly effective than targeting COX or LOX alone, there are only a few reports of examining such compounds in case of colorectal cancers (CRC). In the present work we report that the novel di-tert-butyl phenol-based dual inhibitors DTPSAL, DTPBHZ, DTPINH, and DTPNHZ exhibit significant cytotoxicity against human CRC cell lines. Molecular docking studies revealed a good fit of these compounds in the COX-2 and 5-LOX protein cavities. The inhibitors show significant inhibition of COX-2 and 5-LOX activities and are effective against a panel of human colon cancer cell lines including HCA-7, HT-29, SW480 and intestinal Apc10.1 cells as well as the hyaluronan synthase-2 (Has2) enzyme over-expressing colon cancer cells, through inhibition of the Hyaluronan/CD44v6 cell survival pathway. Western blot analysis and qRT-PCR analyses indicated that the di-tert-butyl phenol-based dual inhibitors reduce the expression of COX-2, 5-LOX, and CD44v6 in human colon cancer HCA-7 cells, while the combination of CD44v6shRNA and DTPSAL has an additional inhibitory effect on CD44v6 mRNA expression. The synergistic inhibitory effect of Celecoxib and Licofelone on CD44v6 mRNA expression suggests that the present dual inhibitors down-regulate cyclooxygenase and lipoxygenase enzymes through CD44v6. The compounds also exhibited enhanced antiproliferative potency compared to standard dual COX/LOX inhibitor, viz. Licofelone. Importantly, the HA/CD44v6 antagonist CD44v6shRNA in combination with synthetic compounds had a sensitizing effect on the cancer cells which enhanced their antiproliferative potency, a finding which is crucial for the anti-proliferative potency of the novel synthetic di-tert-butyl phenol based dual COX-LOX inhibitors in colon cancer cells.
authors.affiliation: Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Shibnath
authors.initials: S
authors.lastname: Ghatak
authors.affiliation: ISTRA Department of Chemistry, Abeda Inamdar Senior College, University of Pune, Pune 411001, India; Department of Bioinformatics and Computer Science, Dr. D.Y. Patil Biotechnology & Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Tathawade, Pune 411033, India.
authors.forename: Alok
authors.initials: A
authors.lastname: Vyas
authors.affiliation: Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA. Electronic address: misra@musc.edu.
authors.forename: Suniti
authors.initials: S
authors.lastname: Misra
authors.affiliation: Hematology/Oncology Division, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: O'Brien
authors.affiliation: Department of Chemistry, Bharati Vidyapeeth, Pune 411007, India.
authors.forename: Ajit
authors.initials: A
authors.lastname: Zambre
authors.affiliation: Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Victor M
authors.initials: VM
authors.lastname: Fresco
authors.affiliation: Regenerative Medicine and Cell Biology, Medical University of South Carolina, Charleston, SC 29425, USA.
authors.forename: Roger R
authors.initials: RR
authors.lastname: Markwald
authors.affiliation: Department of Bioinformatics and Computer Science, Dr. D.Y. Patil Biotechnology & Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Tathawade, Pune 411033, India.
authors.forename: K Venkateshwara
authors.initials: KV
authors.lastname: Swamy
authors.affiliation: Department of Life & Physical Sciences, Lincoln University, 820 Chestnut St., Jefferson City, MO 65101, USA.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Afrasiabi
authors.affiliation: Department of Chemistry, Missouri S & T University (formerly University of Missouri-Rolla), Rolla, MO 65409, USA.
authors.forename: Amitava
authors.initials: A
authors.lastname: Choudhury
authors.affiliation: Department of Bioinformatics and Computer Science, Dr. D.Y. Patil Biotechnology & Bioinformatics Institute, Dr. D.Y. Patil Vidyapeeth, Tathawade, Pune 411033, India.
authors.forename: Madhukar
authors.initials: M
authors.lastname: Khetmalas
authors.affiliation: ISTRA Department of Chemistry, Abeda Inamdar Senior College, University of Pune, Pune 411001, India. Electronic address: sbpadhye@hotmail.com.
authors.forename: Subhash
authors.initials: S
authors.lastname: Padhye
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004354:Drug Screening Assays, Antitumor; D019073:HT29 Cells; D006801:Humans; D006835:Hydrazones / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D016859:Lipoxygenase Inhibitors / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D008958:Models, Molecular; D015394:Molecular Structure; D013329:Structure-Activity Relationship
chemical_list: D000970:Antineoplastic Agents; D006835:Hydrazones; D016859:Lipoxygenase Inhibitors; C012483:phenylhydrazone; D001094:Arachidonate 5-Lipoxygenase; D051546:Cyclooxygenase 2
keywords: Antitumor agents; CD44v6; Colon cancer; Cytotoxicity; Di-tert butyl phenols; Dual COX–LOX inhibitors; Hyaluronan; Molecular modeling