, Volume 22, Issue 1, pp 53–58 | Cite as

Ribonucleases as Novel Chemotherapeutics

The Ranpirnase Example
  • J. Eugene Lee
  • Ronald T. Raines
Novel Therapeutic Strategies


Ranpirnase, a cytotoxic ribonuclease from the frog Rana pipiens, is the archetype of a novel class of cancer chemotherapeutic agents based on homologs and variants of bovine pancreatic ribonuclease (RNase A). Ranpirnase in combination with doxorubicin is in clinical trials for the treatment of unresectable malignant mesothelioma and other cancers. The putative mechanism for ranpirnase-mediated cytotoxicity involves binding to anionic components of the extracellular membrane, cytosolic internalization, and degradation of transfer RNA leading to apoptosis. The maintenance of ribonucleolytic activity in the presence of the cytosolic ribonuclease inhibitor protein is a key aspect of the cytotoxic activity of ranpirnase. The basis for its specific toxicity for cancer cells is not known. This review describes the development of ranpirnase as a cancer chemotherapeutic agent.


Malignant Mesothelioma Conformational Stability Rana Pipiens Cancer Chemotherapeutic Agent Northern Leopard Frog 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Work in the Raines laboratory on ribonucleases is supported by grant CA073808 (NIH). We are grateful to R.F. Turcotte and T.-Y. Chao for comments on the manuscript. The authors have no conflicts of interest that are directly relevant to the content of this review.


  1. 1.
    Tafech A, Bassett T, Sparanese D, et al. Destroying RNA as a therapeutic approach. Curr Med Chem 2006; 13: 863–81PubMedCrossRefGoogle Scholar
  2. 2.
    Sloud M. Ribozymes and siRNAs: from structure to preclinical applications. Handb Exp Pharmacol 2006; 173: 223–42CrossRefGoogle Scholar
  3. 3.
    Schubert S, Kurreck J. Oligonucleotide-based antiviral strategies. Handb Exp Pharmacol 2006; 173: 261–87PubMedCrossRefGoogle Scholar
  4. 4.
    Rayburn ER, Wang H, Zhang R. Antisense-based cancer therapeutics: are we there yet? Expert Opin Emerg Drugs 2006; 11: 337–52PubMedCrossRefGoogle Scholar
  5. 5.
    Roehr B. Forivirsen approved for CMV retinitis. J Int Assoc Physicians AIDS Care 1998; 4: 14–6PubMedGoogle Scholar
  6. 6.
    Bumcrot D, Manoharan M, Koteliansky V, et al. RNAi therapeutics: a potential new class of pharmaceutical drugs. Nat Chem Biol 2006; 2: 711–9PubMedCrossRefGoogle Scholar
  7. 7.
    Aagaard L, Rossi JJ. RNAi therapeutics: principles, prospects and challenges. Adv Drug Deliv Rev 2007; 59: 75–86PubMedCrossRefGoogle Scholar
  8. 8.
    Kim DH, Rossi JJ. Strategies for silencing human disease using RNA interference. Nat Rev Genet 2007; 8: 173–84PubMedCrossRefGoogle Scholar
  9. 9.
    Grimm D, Streetz KL, Jopling CL, et al. Fatality in mice due to oversaturation of cellular microRNA/short hairpin RNA pathways. Nature 2006; 441: 537–41PubMedCrossRefGoogle Scholar
  10. 10.
    Marsden PA. RNA interference as potential therapy: not so fast. New Engl J Med 2006; 355: 953–4PubMedCrossRefGoogle Scholar
  11. 11.
    Matoušek J. Ribonucleases and their antitumor activity. Comp Biochem Physiol 2001; 129C: 175–91Google Scholar
  12. 12.
    Leland PA, Raines RT. Cancer chemotherapy: ribonucleases to the rescue. Chem Biol 2001; 8: 405–13PubMedCrossRefGoogle Scholar
  13. 13.
    Makarov AA, Ilinskaya ON. Cytotoxic ribonucleases: molecular weapons and their targets. FEBS Lett 2003; 540: 15–20PubMedCrossRefGoogle Scholar
  14. 14.
    Benito A, Ribó M, Vilanova M. On the track of antitumor ribonucleases. Mol Biosyst 2005; 1: 294–302PubMedCrossRefGoogle Scholar
  15. 15.
    Arnold U, Ulbrich-Hofmann R. Natural and engineered ribonucleases as potential cancer therapeutics. Biotechnol Lett 2006; 28: 1615–22PubMedCrossRefGoogle Scholar
  16. 16.
    D’Alessio G, Riordan JF, editors. Ribonucleases: structures and functions. New York: cademic Press 1997Google Scholar
  17. 17.
    Raines RT. Ribonuclease A. Chem Rev 1998; 98: 1045–65PubMedCrossRefGoogle Scholar
  18. 18.
    D’Alessio G, Di Donate A, Mazzarella L, et al. Seminal ribonuclease: the importance of diversity. In: D’Alessio G, Riordan JF, editors. Ribonucleases: structures and functions. New York: Academic Press, 1997: 383–423Google Scholar
  19. 19.
    Matoušek J, Souček J, Slavik T, et al. Comprehensive comparison of the cytotoxic activities of Onconase and bovine seminal ribonuclease. Comp Biochem Physiol 2003; 136C: 343–56Google Scholar
  20. 20.
    Costanzi J, Sidransky D, Navon A, et al. Ribonucleases as a novel pro-apoptotic anticancer strategy: review of the preclinical and clinical data for ranpirnase. Cancer Invest 2005; 23: 643–50PubMedCrossRefGoogle Scholar
  21. 21.
    Pavlakis N, Vogelzang NJ. Ranpirnase, an antitumour ribonuclease: its potential role in malignant mesothelioma. Expert Opin Biol Ther 2006; 6: 391–9PubMedCrossRefGoogle Scholar
  22. 22.
    Saxena SK, Shogen K, Ardelt W. Onconase® and its therapeutic potential. Lab Med 2003; 34: 380–7CrossRefGoogle Scholar
  23. 23.
    Shogen K, Yoan WK. Antitumor activity in extracts of Leopard frog (Rana pipiens) embryos. 27th Annual Eastern Colleges Science Conference; 197 Apr 28; State College (PA).Google Scholar
  24. 24.
    Ardelt W, Mikulski SM, Shogen K. Amino acid sequence of an anti-tumor protein from Rana pipiens oocytes and early embryos. J Biol Chem 1991; 266: 245–51PubMedGoogle Scholar
  25. 25.
    Dyer KD, Rosenberg HF. The RNase A superfamily: generation of diversity and innate host defense. Mol Divers 2006; 10: 585–97PubMedCrossRefGoogle Scholar
  26. 26.
    Liao YD, Wang JJ. Yolk granules are the major compartment for bullfrog (Rana catesbeiana) oocyte-specific ribonuclease. Eur J Biochem 1994; 222: 215–20PubMedCrossRefGoogle Scholar
  27. 27.
    Chen S, Le SY, Newton DL, et al. A gender-specific mRNA encoding a cytotoxic ribonuclease contains a 3′ UTR of unusual length and structure. Nucleic Acids Res 2000; 28: 2375–82PubMedCrossRefGoogle Scholar
  28. 28.
    Darzynkiewicz Z, Carter SP, Mikulski SM, et al. Cytostatic and cytotoxic effect of Pannon (P-30 protein), a novel anticancer agent. Cell Tissue Kinet 1988; 21: 169–82PubMedGoogle Scholar
  29. 29.
    Lee I, Kalota A, Gewirtz AM, et al. Antitumor efficacy of the cytotoxic RNase, ranpirnase, on A549 human lung cancer xenografts of nude mice. Anticancer Res 2007; 27: 299–307PubMedGoogle Scholar
  30. 30.
    Mosimann SC, Ardelt W, James MNG. Refined 1.7 Å x-ray crystallographic structure of P-30 protein, an amphibian ribonuclease with anti-tumor activity. J Mol Biol 1994; 236: 1141–53PubMedCrossRefGoogle Scholar
  31. 31.
    Raines RT. Active site of ribonuclease A. In: Zenkova MA, editor. Artificial nucleases. Heidelberg: Springer, 2004: 19–32CrossRefGoogle Scholar
  32. 32.
    Welker E, Hathaway L, Xu G, et al. Oxidative folding and N-terminal cyclization of Onconase. Biochemistry 2007; 46: 5485–93PubMedCrossRefGoogle Scholar
  33. 33.
    Merritt EA, Murphy MEP. Raster3D Version 2.0, a program for photorealistic molecular graphics. Acta Crystallogr D Biol Crystallogr 1994; 50: 869–73PubMedCrossRefGoogle Scholar
  34. 34.
    Lee JE, Raines RT. Contribution of active-site residues to the function of Onconase, a ribonuclease with antitumoral activity. Biochemistry 2003; 42: 11443–50PubMedCrossRefGoogle Scholar
  35. 35.
    Gorbatyuk VY, Tsai CK, Chang CF, et al. Effect of N-terminal and Met23 mutations on the structure and dynamics of Onconase. J Biol Chem 2004; 279: 5772–80PubMedCrossRefGoogle Scholar
  36. 36.
    Merlino A, Mazzarella L, Carannante A, et al. The importance of dynamic effects on the enzyme activity: x-ray structure and molecular dynamics of Onconase mutants. J Biol Chem 2005; 280: 17953–60PubMedCrossRefGoogle Scholar
  37. 37.
    Koshland DE. Application of a theory of enzyme specificity to protein synthesis. Proc Natl Acad Sci USA 1958; 44: 98–104PubMedCrossRefGoogle Scholar
  38. 38.
    Lee JE, Bae E, Bingman CA, et al. Structural basis for catalysis by Onconase. J Mol Biol 2008; 375: 165–77PubMedCrossRefGoogle Scholar
  39. 39.
    Saxena SK, Sirdeshmukh R, Ardelt W, et al. Entry into cells and selective degradation of fRNAs by a cytotoxic member of the RNase A family. J Biol Chem 2002; 277: 15142–6PubMedCrossRefGoogle Scholar
  40. 40.
    Suhasini AN, Sirdeshmukh R. Transfer RNA cleavages by Onconase reveal unusual cleavage sites. J Biol Chem 2006; 281: 12201–9PubMedCrossRefGoogle Scholar
  41. 41.
    Notomista E, Catanzano F, Graziano G, et al. Onconase: an unusually stable protein. Biochemistry 2000; 39: 8711–8PubMedCrossRefGoogle Scholar
  42. 42.
    Leland PA, Staniszewski KE, Kim B-M, et al. A synapomorphic disulfide bond is critical for the conformational stability and cytotoxicity of an amphibian ribonuclease. FEBS Lett 2000; 477: 203–7PubMedCrossRefGoogle Scholar
  43. 43.
    Arnold U, Schulenburg C, Schmidt D, et al. Contribution of structural peculiarities of Onconase to its high stability and folding kinetics. Biochemistry 2006; 45: 3580–7PubMedCrossRefGoogle Scholar
  44. 44.
    Kim B-M, Kim H, Raines RT, et al. Glycosylation of Onconase increases it conformational stability and toxicity for cancer cells. Biochem Biophys Res Commun 2004; 315: 976–83PubMedCrossRefGoogle Scholar
  45. 45.
    Johnson RJ, Chao T-Y, Lavis LD, et al. Cytotoxic ribonucleases: the dichotomy of Coulombic forces. Biochemistry 2007 Sep 11; 46(36): 10308–16PubMedCrossRefGoogle Scholar
  46. 46.
    Wu Y, Mikulski SM, Ardelt W, et al. A cytotoxic ribonuclease: study of the mechanism of Onconase cytotoxicity. J Biol Chem 1993; 268: 10686–93PubMedGoogle Scholar
  47. 47.
    Haigis MC, Raines RT. Secretory ribonucleases are internalized by a dynamin-independent endocytic pathway. J Cell Sci 2003; 116: 313–24PubMedCrossRefGoogle Scholar
  48. 48.
    Rodriguez M, Torrent G, Bosch M, et al. Intracellular pathway of Onconase that enables its delivery to the cytosol. J Cell Sci 2007; 120: 1405–11PubMedCrossRefGoogle Scholar
  49. 49.
    Wu Y, Saxena SK, Ardelt W, et al. A study of the intracellular routing of cytotoxic ribonucleases. J Biol Chem 1995; 270: 17476–81PubMedCrossRefGoogle Scholar
  50. 50.
    Fuchs SM, Raines RT. Internalization of cationic peptides: the road less (or more?) traveled. Cell Mol Life Sci 2006; 63: 1819–22PubMedCrossRefGoogle Scholar
  51. 51.
    Haigis MC, Kurten EL, Raines RT. Ribonuclease inhibitor as an intracellular sentry. Nucleic Acids Res 2003; 31: 1024–32PubMedCrossRefGoogle Scholar
  52. 52.
    Dickson KA, Haigis MC, Raines RT. Ribonuclease inhibitor: structure and function. Prog Nucleic Acid Res Mol Biol 2005; 80: 349–74PubMedCrossRefGoogle Scholar
  53. 53.
    Kajava AV. Structural diversity of leucine-rich repeat proteins. J Mol Biol 1998; 277: 519–27PubMedCrossRefGoogle Scholar
  54. 54.
    Johnson RJ, McCoy JG, Bingman CA, et al. Inhibition of human pancreatic ribonuclease by the human ribonuclease inhibitor protein. J Mol Biol 2007; 367: 434–49CrossRefGoogle Scholar
  55. 55.
    Leland PA, Staniszewski KE, Kim B-M, et al. Endowing human pancreatic ribonuclease with toxicity for cancer cells. J Biol Chem 2001; 276: 43095–102PubMedCrossRefGoogle Scholar
  56. 56.
    Rutkoski TJ, Kurten EL, Mitchell JC, et al. Disruption of shape-complementarity markers to create cytotoxic variants of ribonuclease A. J Mol Biol 2005; 354: 41–54PubMedCrossRefGoogle Scholar
  57. 57.
    Deptala A, Halicka HD, Ardelt B, et al. Potentiation of tumor necrosis factor induced apoptosis by Onconase. Int J Oncol 1998; 13: 11–6PubMedGoogle Scholar
  58. 58.
    Iordanov MS, Ryabinina OP, Wong J, et al. Molecular determinants of apoptosis induced by the cytotoxic ribonuclease Onconase: evidence for cytotoxic mechanisms different from inhibition of protein synthesis. Cancer Res 2000; 60: 1983–94PubMedGoogle Scholar
  59. 59.
    Ardelt B, Ardelt W, Darzynkiewicz Z. Cytotoxic ribonucleases and RNA interference (RNAi). Cell Cycle 2003; 2: 22–4PubMedCrossRefGoogle Scholar
  60. 60.
    Iordanov MS, Wong J, Newton DL, et al. Differential requirement for the stress-activated protein kinase/c-Jun NH2-terminal kinase in RNA damage-induced apoptosis in primary and in immortalized fibroblasts. Mol Cell Biol Res Commun 2000; 4: 122–8PubMedCrossRefGoogle Scholar
  61. 61.
    Grabarek J, Ardelt B, Du L, et al. Activation of caspases and serine proteases during apoptosis induced by Onconase (ranpirnase). Exp Cell Res 2002; 278: 61–71PubMedCrossRefGoogle Scholar
  62. 62.
    Halicka HD, Ardelt B, Shogen K, et al. Mild hyperthermia predisposes tumor cells to undergo apoptosis upon treatment with Onconase. Int J Oncol 2007; 30: 841–7PubMedGoogle Scholar
  63. 63.
    James AM, Ambrose EJ, Lowick JH. Differences between the electrical charge carried by normal and homologous tumour cells. Nature 1956; 177: 576–7PubMedCrossRefGoogle Scholar
  64. 64.
    Slivinsky GG, Hymer WC, Bauer J, et al. Cellular electrophoretic mobility data: a first approach to a database. Electrophoresis 1997; 18: 1109–19PubMedCrossRefGoogle Scholar
  65. 65.
    Kojima K. Molecular aspects of the plasma membrane in tumor cells. Nagoya J Med Sci 1993; 56: 1–18PubMedGoogle Scholar
  66. 66.
    Fredman P. Glycosphingolipid tumor antigens. Adv Lipid Res 1993; 25: 213–34PubMedGoogle Scholar
  67. 67.
    Halicka HD, Murakami T, Papageorgio CN, et al. Induction of differentiation of leukaemic (HL-60) or prostate cancer (LNCaP, JCA-1) cells potentiates apoptosis triggered by Onconase. Cell Prolif 2000; 33: 407–17PubMedCrossRefGoogle Scholar
  68. 68.
    Rybak SM, Pearson JW, Fogler WE, et al. Enhancement of vincristine cytotoxicity in drug-resistant cells by simultaneous treatment with Onconase, an antitumor ribonuclease. J Natl Cancer Inst 1996; 88: 747–53PubMedCrossRefGoogle Scholar
  69. 69.
    Juan G, Ardelt B, Li X, et al. Gl arrest of U937 cells by Onconase is associated with suppression of cyclin D3 expression, induction of p16INK4A, p21WAF1/ CIP1 and p27KIP and decreased pRb phosphorylation. Leukemia 1998; 12: 1241–8PubMedCrossRefGoogle Scholar
  70. 70.
    Mikulski SM, Viera A, Ardelt W, et al. Tamoxifen and trifluoroperazine (Stelazine) potentiate cytostatic/cytotoxic effects of P-30 protein, a novel protein possessing anti-tumor activity. Cell Tissue Kinet 1990; 23: 237–46PubMedGoogle Scholar
  71. 71.
    Mikulski SM, Viera A, Darzynkiewicz Z, et al. Synergism between a novel amphibian oocyte ribonuclease and lovastatin in inducing cytostatic and cytotoxic effects in human lung and pancreatic carcinoma cell lines. Br J Cancer 1992; 66: 304–10PubMedCrossRefGoogle Scholar
  72. 72.
    Lee I, Lee YH, Mikulski SM, et al. Effect of Onconase +/−tamoxifen on ASPC-1 human pancreatic tumors in nude mice. Adv Exp Med Biol 2003; 530: 187–96PubMedCrossRefGoogle Scholar
  73. 73.
    Mikulski SM, Ardelt W, Shogen K, et al. Striking increase of survival of mice bearing M109 Madison carcinoma treated with a novel protein from amphibian embryos. J Natl Cancer Inst 1990; 82: 151–3PubMedCrossRefGoogle Scholar
  74. 74.
    Lee I, Lee YH, Mikulski SM, et al. Enhanced cellular radiation sensitivity of androgen-independent human prostate tumor cells by Onconase. Anticancer Res 2000; 20: 1037–40PubMedGoogle Scholar
  75. 75.
    Mikulski SM, Grossman A, Carter P, et al. Phase I human clinical trial of Onconase (P-30 protein) administered intravenously on a weekly schedule in cancer patients with solid tumors. Int J Oncol 1993; 3: 57–64PubMedGoogle Scholar
  76. 76.
    Mikulski SM, Chun H, Mittelman A, et al. Relationship between response rate and median survival in patients with advanced non-small cell lung cancer: comparison of Onconase with other cancer agents. Int J Oncol 1995; 6: 889–97PubMedGoogle Scholar
  77. 77.
    Puccio C, Mittelman A, Chun H, et al. A new anticancer Rnase (Onconase): clinical trial in patients (pts) with breast cancer (BC) [abstract no. 242]. American Society of Clinical Oncology Annual Meeting; 1996 May 18–21; Philadelphia (PA)Google Scholar
  78. 78.
    Vogelzang NJ, Aklilu M, Stadler WM, et al. A phase II trial of weekly intravenous ranpirnase (Onconase), a novel ribonuclease in patients with metastatic kidney cancer. Invest New Drugs 2001; 19: 255–60PubMedCrossRefGoogle Scholar
  79. 79.
    Mikulski SM, Costanzi JJ, Vogelzang NJ, et al. Phase II trial of a single weekly intravenous dose of ranpirnase in patients with unresectable malignant mesothelioma. J Clin Oncol 2002; 20: 274–81PubMedCrossRefGoogle Scholar
  80. 80.
    Newton DL, Hansen HJ, Mikulski SM, et al. Potent and specific antitumor effects of an anti-CD22-targeted cytotoxic ribonuclease: potential for the treatment of non-Hodgkin lymphoma. Blood 2001; 97: 528–35PubMedCrossRefGoogle Scholar

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© Adis Data Information BV 2008

Authors and Affiliations

  1. 1.Department of BiochemistryUniversity of Wisconsin-MadisonMadisonUSA
  2. 2.Division of BiologyCalifornia Institute of TechnologyPasadenaUSA
  3. 3.Department of ChemistryUniversity of Wisconsin-MadisonMadisoncUSA

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