Effect of Wf-536, a novel ROCK inhibitor, against metastasis of B16 melanoma

Cancer Chemother Pharmacol. 2003 Oct;52(4):319-24. doi: 10.1007/s00280-003-0641-9. Epub 2003 May 29.

Abstract

Purpose: Rho-associated coiled-coil-forming protein kinase (ROCK) is pivotally involved in invasion by tumor cells and their evolution to metastasis. We have developed a novel inhibitor of ROCK, Wf-536 [(+)-(R)-4-(1-aminoethyl)-N-(4-pyridyl) benzamide monohydrochloride]. In the present study, we investigated its effect on in vitro invasion and in vivo pulmonary metastasis of B16 melanoma.

Methods: The following were evaluated: the anti-invasive effect of Wf-536 against the motility of mouse B16BL6 melanoma cells through a culture insert layered with reconstituted basement membrane (Matrigel); the cytotoxic effect of Wf-536 in the same cell line; the antimetastatic effect of Wf-536, administered by osmotic pump, on spontaneous pulmonary metastasis following subcutaneous injection of B16BL6 melanoma in mice; and the inhibitory effect of orally administered Wf-536, alone or in combination with the antineoplastic drug paclitaxel, on pulmonary metastasis of intravenously injected B16F10 melanoma in mice.

Results: Wf-536 inhibited in vitro invasion by B16BL6 cells significantly and in a concentration-dependent manner and displayed an anti-invasive effect under conditions of both chemotaxis and chemokinesis. No cytotoxic effect was observed at any of the concentrations used. In vivo, Wf-536 administration suppressed tumor colony formation on the lung surface in a dose-dependent manner (0.3-3 mg/kg per day), with a metastasis inhibition rate of 95% at 3 mg/kg per day. In experimental metastasis of B16F10 melanoma, oral administration of Wf-536 significantly decreased tumor colony formation in the lung, with an inhibition rate of 41% at 3 mg/kg per day. The inhibition rate of paclitaxel (5 mg/kg per day) was 27%. The combination of Wf-536 and paclitaxel produced a synergistic effect on B16F10 metastasis and a 68% inhibition rate. Wf-536 administration at the doses used did not alter body weight, blood pressure or the health of treated animals as compared to vehicle-treated controls.

Conclusion: The results suggest that Wf-536 is a potentially valuable drug for preventing tumor metastasis both in monotherapy and in combination with an antineoplastic drug.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology*
  • Antineoplastic Agents, Phytogenic / therapeutic use
  • Benzamides / pharmacology*
  • Cell Line, Tumor
  • Cell Movement / drug effects
  • Cell Survival / drug effects
  • Enzyme Inhibitors / pharmacology*
  • Intracellular Signaling Peptides and Proteins
  • Lung Neoplasms / drug therapy*
  • Lung Neoplasms / secondary*
  • Melanoma, Experimental / drug therapy*
  • Melanoma, Experimental / pathology*
  • Mice
  • Neoplasm Invasiveness / prevention & control
  • Paclitaxel / therapeutic use
  • Protein-Serine-Threonine Kinases / antagonists & inhibitors*
  • Pyridines / pharmacology*
  • rho-Associated Kinases

Substances

  • Antineoplastic Agents
  • Antineoplastic Agents, Phytogenic
  • Benzamides
  • Enzyme Inhibitors
  • Intracellular Signaling Peptides and Proteins
  • Pyridines
  • Y 32885
  • Protein-Serine-Threonine Kinases
  • rho-Associated Kinases
  • Paclitaxel