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Arginine Deprivation Therapy Via Arginine Deiminase

Arginine Deprivation Therapy Via Arginine Deiminase

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Arginine Deprivation Therapy Via Arginine Deiminase

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  1. Arginine Deprivation Therapy Via Arginine Deiminase Mary Rose Stevens CHM 357, Smith College October 31, 2007

  2. Outline • General topic: cancer treatment • Malignant melanoma, hepatocellular carcinoma • Cell biology review • urea cycle • argininosuccinate synthetase (ASS) • Arginine deiminase (ADI) • discovery of anticancer effects • arginine auxotrophy • ADI structure and mechanism

  3. Outline • Eligibility for ADI treatment • ASS expression in cancer cell lines • Resistance to arginine deprivation • Drug design • ADI-PEG 20 • Phase I/II trials • Current status of ADI-PEG 20 • Companies, clinical trials • Conclusion

  4. Malignant melanoma and HCC • Kill most within 1 year of diagnosis • 16,000 deaths/yr in the U.S. • Melanoma is rapidly increasing • Australia • Urgent need for effective treatments • Amino acid deprivation

  5. The Urea Cycle

  6. Argininosuccinate Synthetase Assumed biological molecule (left) and assymetric molecule (right) from source, E. coli

  7. Mycoplasma • 2004: Ensor et al found that Mycoplasma kills cervical carcinoma and leukemia cell lines • Mycoplasma contains the arginine-degrading enzyme, arginine deiminase (ADI)

  8. Arginine Deiminase (ADI) • ADI (Psuedomonas aeruginosa) in complex with a reaction intermediate (left) • ADI (Mycoplasma arginini) in complex with arginine (right)

  9. ADI Enzymatic Action

  10. Arginine to Citrulline and Ammonia via ADI

  11. ASS Expression in Cancer Cell Lines • In 2007, Savaraj et al examined ASS expression levels in melanoma cell lines (A375, Sk-Mel-2, A2058, MEL-1220) • Also examined ASS DNA levels; lack of ASS expression is not from gene deletion

  12. Pegylated Arginine Deiminase (ADI-PEG20) • Problem: ADI has a short circulating half-life and high antigenicity • Solution: Polaris, Inc. developed a pegylated ADI (ADI-PEG 20), which has shown promise in clinical trials with melanoma patients

  13. Resistance to Treatment • ASS expression in 4 melanoma cell lines before and after ADI-PEG20 treatment In current

  14. Phase I/II: HCC • Auxotrophic for arginine • ADI-PEG 20 is well-tolerated • OBD: 160 U/m2 • IM injection, once weekly • Reasonable anti-tumor response rate • Prolonged survival

  15. Phase I/II: Stage IV Melanoma • Auxotrophic for arginine • ADI-PEG 20 is well-tolerated • OBD: 160 U/m2 • IM injection, once weekly • Reasonable anti-tumor response rate (25%) • Prolonged survival (mean and median)

  16. Inhibition of NO Production • Nitric oxide synthase (NOS) catalyzes nitric oxide (NO) production from extracellular arginine • ADI-PEG 20 inhibited NO production • No increased b.p. or h.r. • Additional mechanisms of action

  17. Current Status of ADI-PEG 20 • Phoenix Pharmacologics, Inc. • Polaris Pharmaceuticals, Inc. • Complete • Phase I, malignant melanoma (U.S.) • Phase I and II, malignant melanoma (Italy) • Phase I and II, HCC (U.S.) • Phase I and II, HCC (Italy) • In Progress • Phase II, malignant melanoma (U.S.) • Phase IIB / III, HCC (Italy)

  18. Current Status of ADI-PEG 20 • No human toxicities observed to date • Medical benefit observed in HCC • Primary market • Metastatic melanoma - US (14000), EU (15000), AUS (15000) • HCC - US (14000), EU (16000), Japan (125000)

  19. Conclusion • ADI-PEG 20 • For cancer cell lines auxotrophic for arginine • Effective in lowering plasma arginine • Well tolerated • Appears to have anti-tumor activity and prolong survival of patients with melanoma and HCC • Multiple mechanisms

  20. References • ADI-PEG 20. Polaris. Online: http://www.polarispharma.com/ADI-PEG%2020.htm. Accessed October 22, 2007. • ADI-PEG 20- Arginine deiminase conjugated with poly(ethylene) glycol. Phoenix Pharmacologics, Inc. Online: http://www.phoenixpharmaco.com/ADIPEG20.htm. Accessed October 22, 2007. • Ascierto, P. A.; Scala, S.; Castello, G.; Daponte, A.; Simeone, E.; Ottaiano, A.; Beneduce, G.; De Rosa, V.; Izzo, F.; Melucci, M. T.; Ensor, C. M.; Prestayko, A. W.; Holtsberg, F. W.; Bomalaski, J. S.; Clark, M. A.; Savaraj, N.; Feun, L. G.; Logan, T. F. Pegylated arginine deiminase treatment of patients with metastatic melanoma: results from phase I and II studies. Journal of Clinical Oncology. 2005, 23, 7660-7668. • Clark, M. A. European Patent EP1337630, 2003. • Dillon, B.; Prieto, V. G.; Curley, S. A.; Ensor, M. C.; Holtsberg, F. W.; Bomalaski, J. S.; Clark, M. A. Incidence and distribution of argininosuccinate synthetase deficiency in human cancers: a method for identifying cancers sensitive to arginine deprivation. Cancer.2004, 100, 826-833. • Durzan. Figure 1: The reactions of the Krebs-Henseleit (urea) cycle, and their relations to NOS activity. BMC Plant Biology. 2005, 5, S12. Online: http://www.biomedcentral.com/1471-2229/5/S1/S12/figure/F1. Accessed October 23, 2007. • Galkin, A.; Kulakova, L.; Sarikaya, E.; Lim, K.; Howard, A.; Herzberg, O. Structural insight into arginine degradation by arginine deiminase, an antibacterial and parasitic drug target. Journal of Biological Chemistry.2004, 279, 14001-14008. • Hardy Diagnostics. Microscopic image of Mycoplasma hominis and Ureaplasma urealyticum colonies growing on A8 Agar. Online: http://www.hardydiagnostics.com/catalog2/hugo/A8Agar.htm. Accessed October23, 2007. • Husson, A.; Brasse-Lagnel, C.; Fairand, A.; Renouf, S.; Lavoinne, A. Argininosuccinate synthetase from the urea cycle to the citrulline-NO cycle. Eur. J. Biochem.2003, 270, 1887-1899. • L-arginine iminohydrolase. Kyoto Encyclopedia of Genes and Genomes. Online: http://www.genome.ad.jp/dbget-bin/www_bget?reaction+R00552+R06138. Accessed October 23, 2007. • PBD ID: 1K92. Lemke, C.T.; Howell, P.L.; The 1.6 Å crystal structure of E. coli argininosuccinate synthetase suggests a conformational change during catalysis. Structure. 2001, 9, 1153-1164. • PBD ID: 1SIR. Das, K.; Buttler, G. H; Kwiatkowski, V.; Clark Jr., A. D.; Yadav, P.; Arnold, E. Crystal structures of arginine deiminase with covalent reaction intermediates: implications for catalytic mechanism. Structure. 2004,12, 657-667. • PBD ID: 2A9G. Galkin, A.; Lu, X.; Dunaway-Mariano, D.; Herzberg, O. Crystal structures representing the Michaelis complex and the thiouronium reaction intermediate of Pseudomonas aeruginosa arginine deiminase. J. Biol. Chem.2005, 280, 34080-34087. • Savaraj, N.; Wu, C.; Kuo, M. T.; You, M.; Wangpaichitr, M.; Robles, C.; Spector, S.; Feun, L. The relationship of arginine deprivation, argininosuccinate synthetase and cell death in melanoma. Drug Target Insights. 2007, 2, 119-128.