title: Reversible action of diaminothiazoles in cancer cells is implicated by the induction of a fast conformational change of tubulin and suppression of microtubule dynamics.
abstract: Diaminothiazoles are novel cytotoxic compounds that have shown efficacy toward different cancer cell lines. They show potent antimitotic and antiangiogenic activity upon binding to the colchicine-binding site of tubulin. However, the mechanism of action of diaminothiazoles at the molecular level is not known. Here, we show a reversible binding to tubulin with a fast conformational change that allows the lead diaminothiazole DAT1 [4-amino-5-benzoyl-2-(4-methoxy phenyl amino)thiazole] to cause a reversible mitotic block. DAT1 also suppresses microtubule dynamic instability at much lower concentration than its IC(50) value in cancer cells. Both growth and shortening events were reduced by DAT1 in a concentration-dependent way. Colchicine, the long-studied tubulin-binding drug, has previously failed in the treatment of cancer due to its toxicity, even though it generates a strong apoptotic response. The toxicity is attributable to its slow removal from the cell due to irreversible tubulin binding caused by a slow conformational change. DAT1 binds to tubulin at an optimal pH lower than colchicine. Tubulin conformational studies showed that the binding environments of DAT1 and colchicine are different. Molecular dynamic simulations showed a difference in the number of H-bonding interactions that accounts for the different pH optima. This study gives an insight of the action of compounds targeting tubulin's colchicine-binding site, as many such compounds have entered into clinical trials recently.
authors.affiliation: Corresponding Author: Suparna Sengupta, Division of Cancer Research, Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram 695014, India. ssengupta@rgcb.res.in.
authors.forename: Nisha E
authors.initials: NE
authors.lastname: Thomas
authors.forename: Reshma
authors.initials: R
authors.lastname: Thamkachy
authors.forename: Krishnankutty C
authors.initials: KC
authors.lastname: Sivakumar
authors.forename: Krishnakumar J
authors.initials: KJ
authors.lastname: Sreedevi
authors.forename: Xavier Lieben
authors.initials: XL
authors.lastname: Louis
authors.forename: Sannu A
authors.initials: SA
authors.lastname: Thomas
authors.forename: Rohith
authors.initials: R
authors.lastname: Kumar
authors.forename: Kallikat N
authors.initials: KN
authors.lastname: Rajasekharan
authors.forename: Lynne
authors.initials: L
authors.lastname: Cassimeris
authors.forename: Suparna
authors.initials: S
authors.lastname: Sengupta
mesh_terms: D049109:Cell Proliferation / Q000187:drug effects; D003078:Colchicine / Q000008:administration & dosage; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D045325:HCT116 Cells; D006801:Humans; D006860:Hydrogen Bonding; D008870:Microtubules / Q000187:drug effects*; D008938:Mitosis / Q000187:drug effects; D056004:Molecular Dynamics Simulation; D011487:Protein Conformation / Q000187:drug effects*; D013844:Thiazoles / Q000008:administration & dosage*; D014404:Tubulin / Q000737:chemistry / Q000187:drug effects*
chemical_list: C496402:4-amino-5-benzoyl-2-(4-methoxyphenylamino)thiazole; D013844:Thiazoles; D014404:Tubulin; D003078:Colchicine