Skip to main content

Advertisement

SpringerLink
Log in

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

  • Original Article
  • Published:
Cancer Chemotherapy and Pharmacology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2A, B.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.

Similar content being viewed by others

References

  1. Belotti D, Vergani V, Drudis T, Borsotti P, Pitelli MR, Viale G, Giavazzi R, Taraboletti G (1996) The microtubule-affecting drug paclitaxel has antiangiogenic activity. Clin Cancer Res 2:1843

    CAS  PubMed  Google Scholar 

  2. Blagosklonny MV, Chuman Y, Bergan RC, Fojo T (1999) Mitogen-activated protein kinase pathway is dispensable for microtubule-active drug-induced Raf-1/Bcl-2 phosphorylation and apoptosis in leukemia cells. Leukemia 13:1028

    Article  CAS  PubMed  Google Scholar 

  3. Chirivi RG, Garofalo A, Crimmin MJ, Bawden LJ, Stoppacciaro A, Brown PD, Giavazzi R (1994) Inhibition of the metastatic spread and growth of B16-BL6 murine melanoma by a synthetic matrix metalloproteinase inhibitor. Int J Cancer 58:460

    Google Scholar 

  4. Ferrante K, Winograd B, Canetta R (1999) Promising new developments in cancer chemotherapy. Cancer Chemother Pharmacol [Suppl] 43:S61

    Google Scholar 

  5. Fidler IJ (1975) Biological behavior of malignant melanoma cells correlated to their survival in vivo. Cancer Res 35:218

    CAS  PubMed  Google Scholar 

  6. Fujii A, Otsuki M, Kataoka H, Chiba K, Fujita F, Koike M, Fujita M, Sakamoto Y (2001) Wf-536, a novel and potent inhibitor of Rho-associated protein kinase (ROCK): inhibitory effects on lymph node and pulmonary metastasis of mouse B16-F10 melanoma. Proc Am Assoc Cancer Res 42:924

    Google Scholar 

  7. Haldar S, Basu A, Croce CM (1997) Bcl-2 is the guardian of microtubule integrity. Cancer Res 57:229

    CAS  PubMed  Google Scholar 

  8. Hart IR (1979) The selection and characterization of an invasive variant of the BL6 melanoma. Am J Pathol 97:587

    CAS  PubMed  Google Scholar 

  9. Hayashi K, Amano Y, Katayama K, Nakajima M, Egi Y, Uehata M, Arii S, Imamura M, Goto N (2001) Wf-536, a novel and potent inhibitor of Rho-associated protein kinase (ROCK): inhibition of in vitro tumor cell invasion and spontaneous evolution of mouse tumors into metastasis. Proc Am Assoc Cancer Res 42:924

    Google Scholar 

  10. Imamura F, Mukai M, Ayaki M, Akedo H (2000) Y-27632, an inhibitor of Rho-associated protein kinase, suppresses tumor cell invasion via regulation of focal adhesion and focal adhesion kinase. Jpn J Cancer Res 91:811

    CAS  PubMed  Google Scholar 

  11. Inoue K, Slaton JW, Perrotte P, Davis DW, Bruns CJ, Hicklin DJ, McConkey DJ, Sweeney P, Radinsky R, Dinney CP (2000) Paclitaxel enhances the effects of the anti-epidermal growth factor receptor monoclonal antibody ImClone C225 in mice with metastatic human bladder transitional cell carcinoma. Clin Cancer Res 6:4874

    CAS  PubMed  Google Scholar 

  12. Itoh K, Yoshioka K, Akedo H, Uehata M, Ishizaki T, Narumiya S (1999) An essential part for Rho-associated kinase in the transcellular invasion of tumor cells. Nat Med 5:221

    CAS  PubMed  Google Scholar 

  13. Kahan BD (1995) Concentration-controlled immunosuppressive regimens using cyclosporine with sirolimus or brequinar in human renal transplantation. Transplant Proc 27:33

    Google Scholar 

  14. Kalechman Y, Longo DL, Catane R, Shani A, Albeck M, Sredni B (2000) Synergistic anti-tumoral effect of paclitaxel (Taxol)+AS101 in a murine model of B16 melanoma: association with ras-dependent signal-transduction pathways. Int J Cancer 86:281

    Google Scholar 

  15. Ling V, Chambers AF, Harris JF, Hill RP (1985) Quantitative genetic analysis of tumor progression. Cancer Metastasis Rev 4:173

    CAS  PubMed  Google Scholar 

  16. Locker J, Goldblatt PJ, Leighton J (1970) Ultrastructural features of invasion in chick embryo liver metastasis of Yoshida ascites hepatoma. Cancer Res 30:1632

    CAS  PubMed  Google Scholar 

  17. Miller KD, Sweeney CJ, Sledge GW Jr (2001) Redefining the target: chemotherapeutics as antiangiogenics. J Clin Oncol 19:1195

    CAS  PubMed  Google Scholar 

  18. Mohler JL, Partin AW, Coffey DS (1987) Prediction of metastatic potential by a new grading system of cell motility: validation in the Dunning R-3327 prostatic adenocarcinoma model. J Urol 138:168

    CAS  PubMed  Google Scholar 

  19. Nakajima M, Katayama K, Tamechika I, Hayashi K, Amano Y, Uehata M, Goto N (2001) Wf-536, a novel and potent inhibitor of Rho-associated protein kinase (ROCK): inhibition of in vitro processes leading to tumor cell invasion and vascularization. Proc Am Assoc Cancer Res 42:924

    Google Scholar 

  20. Neri A, Goggin B, Kolis S, Brekken J, Khelemskaya N, Gabriel L, Robinson SR, Webber S, Wood AW, Appelt K, Shalinsky DR (1998) Pharmacokinetics and efficacy of a novel matrix metalloproteinase inhibitor, AG3340, in single agent and combination therapy against B16-F10 melanoma tumors developing in lung after IV-tail implantation in C57BL/6 mice. Proc Am Assoc Cancer Res 39:2060

    Google Scholar 

  21. Partin AW, Mohler JL, Coffey DS (1992) Cell motility as an index of metastatic ability in prostate cancers: results with an animal model and with human cancer cells. Cancer Treat Res 59:121

    CAS  PubMed  Google Scholar 

  22. Rowinski EK (1994) Update on the antitumor activity of paclitaxel in clinical trials. Ann Pharmacother 28: S18

    PubMed  Google Scholar 

  23. Schiff PB, Horwitz SB (1980) Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A 77:1561

    CAS  PubMed  Google Scholar 

  24. Shalinsky DR, Brekken J, Zou H, Mcdermott CD, Forsyth P, Edwards D, Margosiak S, Bender S, Truitt G, Wood A, Varki NM, Appelt K (1999) Broad antitumor and antiangiogenic activities of AG3340, a potent and selective MMP inhibitor undergoing advanced oncology clinical trials. In: Greenwald RA, Zucker S, Golub LM (ed) Inhibition of matrix metalloproteinases: therapeutic applications. Ann N Y Acad Sci 878:236

    CAS  PubMed  Google Scholar 

  25. Somlyo AV, Bradshaw D, Ramos S, Murphy C, Myers CE, Somlyo AP (2000) Rho-kinase inhibitor retards migration and in vivo dissemination of human prostate cancer cells. Biochem Biophys Res Commun 269:652

    CAS  PubMed  Google Scholar 

  26. Watanabe K, Ito S, Kikuchi K, Ichikawa N, Ando Y, Meigata K, Nomura Y, Degata T, Beck Y, Tomikawa S, Nagao T, Uchida H (1996) Synergistic effect of tacrolimus and mizoribine on in vitro and in vivo experiments assessed by a combination index. Jap J Transplant 31:23

    Google Scholar 

  27. Wyckoff JB, Jones JG, Condeelis JS, Segall JE (2000) A critical step in metastasis: in vivo analysis of intravasation at the primary tumor. Cancer Res 60:2504

    CAS  PubMed  Google Scholar 

  28. Zagozdzon R, Golab J, Mucha K, Foroncewicz B, Jakobisiak M (1999) Potentiation of antitumor effects of IL-12 in combination with paclitaxel in murine melanoma model in vivo. Int J Mol Med 4:645

    Google Scholar 

Download references

Acknowledgements

The authors would like to thank Dr Y. Matsumoto and Mrs. A. Ohno for their helpful comments during the preparation of the manuscript.

Author information

Authors and Affiliations

  1. Pharmaceuticals Research Unit, Mitsubishi Pharma Corporation, 1000, Kamoshida-cho, Aoba-ku, 227-0033, Yokohama, Japan

    Masahide Nakajima, Kazutaka Hayashi, Yasuhiro Egi, Ken-ichi Katayama, Yusaku Amano, Masayoshi Uehata, Makio Ohtsuki, Akihiro Fujii, Koh-ichi Oshita, Hirotoshi Kataoka, Kenji Chiba, Nobuharu Goto & Takao Kondo

Authors
  1. Masahide Nakajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kazutaka Hayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yasuhiro Egi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ken-ichi Katayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yusaku Amano
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masayoshi Uehata
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Makio Ohtsuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Akihiro Fujii
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Koh-ichi Oshita
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Hirotoshi Kataoka
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Kenji Chiba
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Nobuharu Goto
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Takao Kondo
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Masahide Nakajima.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nakajima, M., Hayashi, K., Egi, Y. et al. Effect of Wf-536, a novel ROCK inhibitor, against metastasis of B16 melanoma. Cancer Chemother Pharmacol 52, 319–324 (2003). https://doi.org/10.1007/s00280-003-0641-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00280-003-0641-9

Keywords

Search

Navigation