Novel derivatives of 1,3,4-oxadiazoles are potent mitostatic agents featuring strong microtubule depolymerizing activity in the sea urchin embryo and cell culture assays.

A series of novel 1,3,4-oxadiazole derivatives based on structural and electronic overlap with combretastatins have been designed and synthesized. Initially, we tested all new compounds in vivo using the phenotypic sea urchin embryo assay to yield a number of agents with anti-proliferative, anti-mitotic, and microtubule destabilizing activities. The experimental data led to identification of 1,3,4-oxadiazole derivatives with isothiazole (5-8) and phenyl (9-12) pharmacophores featuring activity profiles comparable to that of combretastatins, podophyllotoxin and nocodazole. Cytotoxic effects of the two lead molecules, namely 6 and 12, were further confirmed and evaluated by conventional assays with the A549 human cancer cell line including cell proliferation, cell cycle arrest at the G2/M phase, cellular microtubule distribution, and finally in vitro microtubule assembly with purified tubulin. The modeling results using 3D similarity (ROCS) and docking (FRED) correlated well with the observed activity of the molecules. Docking data suggested that the most potent molecules are likely to target the colchicine binding site.

[1]  J. Sloan,et al.  Phase I trial of dolastatin-10 (NSC 376128) in patients with advanced solid tumors. , 1999, Clinical cancer research : an official journal of the American Association for Cancer Research.

[2]  Alan J. Lacey.  Light microscopy in biology , 1989 .

[3]  R. Donehower,et al.  Neurotoxicity of Taxol. , 1993, Journal of the National Cancer Institute. Monographs.

[4]  Dennis Bray,et al.  Cell Movements: From Molecules to Motility , 1992 .

[5]  R. C.-Gaudreault,et al.  N-Phenyl-N'-(2-chloroethyl)urea analogues of combretastatin A-4: Is the N-phenyl-N'-(2-chloroethyl)urea pharmacophore mimicking the trimethoxy phenyl moiety? , 2007, Bioorganic & medicinal chemistry letters.

[6]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[7]  N. Lawrence,et al.  Tubulin as a Target for Anticancer Drugs: Agents which Interact with the Mitotic Spindle , 1998 .

[8]  T. Fojo,et al.  Paclitaxel-resistant Human Ovarian Cancer Cells Have Mutant β-Tubulins That Exhibit Impaired Paclitaxel-driven Polymerization* , 1997, The Journal of Biological Chemistry.

[9]  Y. Engelborghs,et al.  Molecular Recognition of Taxol by Microtubules , 2000, The Journal of Biological Chemistry.

[10]  S A Kuznetsov,et al.  Myosin Va bound to phagosomes binds to F-actin and delays microtubule-dependent motility. , 2001, Molecular biology of the cell.

[11]  A. Lacey,et al.  Light microscopy in biology : a practical approach , 1989 .

[12]  M. Jordan,et al.  Microtubules as a target for anticancer drugs , 2004, Nature Reviews Cancer.

[13]  A. Bershadsky,et al.  High molecular weight protein MAP 2 promoting microtubule assembly in vitro is associated with microtubules in cells. , 1980, Cell biology international reports.

[14]  Giovanni Sorba,et al.  Medicinal chemistry of combretastatin A4: present and future directions. , 2006, Journal of medicinal chemistry.

[15]  G. M. Wilson,et al.  Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells. , 1994, Cancer research.

[16]  K. Gewald,et al.  Synthese und Reaktionen von 4‐Aminoisothiazolen , 1979 .

[17]  M. Kavallaris,et al.  Taxol-resistant epithelial ovarian tumors are associated with altered expression of specific beta-tubulin isotypes. , 1997, The Journal of clinical investigation.

[18]  S Seeber,et al.  D-24851, a novel synthetic microtubule inhibitor, exerts curative antitumoral activity in vivo, shows efficacy toward multidrug-resistant tumor cells, and lacks neurotoxicity. , 2001, Cancer research.

[19]  S. Terakawa.  Video Microscopy , 1985, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[20]  Anurag Chaudhary,et al.  Combretastatin a-4 analogs as anticancer agents. , 2007, Mini reviews in medicinal chemistry.

[21]  D. Zaharevitz,et al.  A common pharmacophore for a diverse set of colchicine site inhibitors using a structure-based approach. , 2005, Journal of medicinal chemistry.

[22]  A. Kiselyov,et al.  Sea urchin embryo as a model organism for the rapid functional screening of tubulin modulators. , 2006, BioTechniques.

[23]  David G I Kingston.  Tubulin-interactive natural products as anticancer agents. , 2009, Journal of natural products.

[24]  N. Nam.  Combretastatin A-4 analogues as antimitotic antitumor agents. , 2003, Current medicinal chemistry.

[25]  M. Molodtsov,et al.  Research Article: In vivo Evaluation of Indolyl Glyoxamides in the Phenotypic Sea Urchin Embryo Assay , 2007, Chemical biology & drug design.

[26]  Patrick A. Curmi,et al.  Insight into tubulin regulation from a complex with colchicine and a stathmin-like domain , 2004, Nature.

[27]  E. Hamel,et al.  Interactions of tubulin with potent natural and synthetic analogs of the antimitotic agent combretastatin: a structure-activity study. , 1988, Molecular pharmacology.

[28]  Eric Deniau,et al.  First synthesis and pharmacological evaluation of benzoindolizidine and benzoquinolizidine analogues of α- and β-peltatin , 2000 .

[29]  T. Mitchison,et al.  Dynamics of microtubule assembly in vivo , 1994 .

[30]  E. Nogales,et al.  High-Resolution Model of the Microtubule , 1999, Cell.

[31]  A. Malikzay,et al.  Oxadiazole derivatives as a novel class of antimitotic agents: Synthesis, inhibition of tubulin polymerization, and activity in tumor cell lines. , 2006, Bioorganic & medicinal chemistry letters.

[32]  J. A. Grant,et al.  Gaussian docking functions. , 2003, Biopolymers.

[33]  A. Malikzay,et al.  Synthesis and structure-activity relationships of 1,2,4-triazoles as a novel class of potent tubulin polymerization inhibitors. , 2005, Bioorganic & medicinal chemistry letters.

[34]  M. Boyd,et al.  Antineoplastic agents. 291. Isolation and synthesis of combretastatins A-4, A-5, and A-6(1a) , 1995, Journal of medicinal chemistry.

[35]  D. Panda,et al.  Modulation of microtubule dynamics by drugs: a paradigm for the actions of cellular regulators. , 1999, Cell structure and function.

[36]  E. Nogales,et al.  Refined structure of alpha beta-tubulin at 3.5 A resolution. , 2001, Journal of molecular biology.

[37]  P. Hawkins,et al.  Comparison of shape-matching and docking as virtual screening tools. , 2007, Journal of medicinal chemistry.

[38]  K. Balakin,et al.  Recent progress in discovery and development of antimitotic agents. , 2007, Anti-cancer agents in medicinal chemistry.

[39]  R. Flavell,et al.  Double knockout of the MRP gene leads to increased drug sensitivity in vitro. , 1996, Cancer research.

[40]  E. Hamel.  Antimitotic natural products and their interactions with tubulin , 1996, Medicinal research reviews.

[41]  S. Giorgi-Renault,et al.  Drugs that inhibit tubulin polymerization: the particular case of podophyllotoxin and analogues. , 2012, Current medicinal chemistry. Anti-cancer agents.