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Investigational New Drugs

, Volume 32, Issue 5, pp 795–805 | Cite as

Pharmacology, immunogenicity, and efficacy of a novel pegylated recombinant Erwinia chrysanthemi-derived L-asparaginase

  • Wei-Wen Chien
  • Soraya Allas
  • Nicolas Rachinel
  • Pierre Sahakian
  • Michel Julien
  • Céline Le Beux
  • Claire-Emmanuelle Lacroix
  • Thierry Abribat
  • Gilles SallesEmail author
PRECLINICAL STUDIES

Summary

Bacterial L-asparaginase (ASNase), hydrolyzing L-asparagine (Asn), is an indispensable component used in the treatment of acute lymphoblastic leukemia (ALL) and certain lymphoma entities. Native Erwinia chrysanthemi-derived ASNase (n-crisantaspase) has been approved as a second-line drug for treating patients exhibiting allergy syndromes to native and pegylated Escherichia coli-derived ASNase (EC-ASNase). However, it still induces hypersensitivity in at least 17 % of treated patients. In the present study, we investigated the pharmacological activity, immunogenicity and anti-leukemic activity of a new pegylated recombinant crisantaspase (PEG-r-crisantaspase). The results demonstrate that when compared to n-crisantaspase in vivo, PEG-r-crisantaspase maintains a complete depletion of plasma Asn for up to 72 h with a 50-fold lower dose. In mice receiving PEG-r-crisantaspase, specific antibodies against the enzyme were undetectable, indicating a lower immunogenicity of the pegylated enzyme. In vitro, PEG-r-crisantaspase exhibits similar cytotoxic effects (EC50 < 5 × 10−4 U/mL for the most sensitive cell lines) to n-crisantaspase on various leukemia and lymphoma cells and was shown to be more efficient than EC-ASNase. Three repeated PEG-r-crisantaspase injections (2–20 U/Kg) prevented leukemia development in leukemia-bearing mice for 17 days and significantly prolonged animal survival to 7–12 days. Therefore, PEG-r-crisantaspase appears to be a promising drug candidate for ALL treatment and should be further explored in experimental and clinical trials.

Keywords

Acute lymphoblastic leukemia Lymphoma L-asparaginase Erwinia chrysanthemi pegylation 

Notes

Acknowledgments

Special thanks to Dr. Philippe Gaulard (INSERM U955, France), Dr. Paul Coppo (Hôpital Saint-Antoine, Paris, France), and Dr. Hiroshi Kimura (Department of Virology, Japan) for providing MEC04 and KHYG1 cells lines, respectively. Sincere thanks to Dr. Christine Saban for amino acid measurement, Patrick Manas for his help in animal experiments, Martine Ffrench and Aline Billoud for patient samples, Lucile Baseggio for the immunocytochemistry technique, and to the ALL patients who donated blood samples for our studies. The project received partial financial support with a refundable grant from Oséo (Maisons Alfort, France) and an FUI (Fonds Unique Interministériel) grant contributed by both the European Regional Development Fund (ERDF), and the Grand Lyon and Rhône Alpes regions.

Conflict of interest

SA PS, MJ, and TA are employees, and TA is a shareholder of Alizé Pharma II. The remaining authors declare no competing financial interests.

Supplementary material

10637_2014_102_MOESM1_ESM.doc (54 kb)
Supplemental Table 1 (DOC 54 kb)
10637_2014_102_MOESM2_ESM.doc (764 kb)
Supplemental Figure 1 (DOC 764 kb)

References

  1. 1.
    Pieters R, Hunger SP, Boos J, Rizzari C, Silverman L, Baruchel A, Goekbuget N, Schrappe M, Pui CH (2011) L-asparaginase treatment in acute lymphoblastic leukemia: a focus on Erwinia asparaginase. Cancer 117(2):238–249PubMedCentralCrossRefPubMedGoogle Scholar
  2. 2.
    Asselin BL (1999) The three asparaginases. Comparative pharmacology and optimal use in childhood leukemia. Adv Exp Med Biol 457:621–629CrossRefPubMedGoogle Scholar
  3. 3.
    Asselin BL (2012) The right dose for the right patient. Blood 119(7):1617–1618CrossRefPubMedGoogle Scholar
  4. 4.
    Avramis VI (2011) Asparaginases: a successful class of drugs against leukemias and lymphomas. J Pediatr Hematol Oncol 33(8):573–579CrossRefPubMedGoogle Scholar
  5. 5.
    Kidd JG (1953) Regression of transplanted lymphomas induced in vivo by means of normal guinea pig serum. I. Course of transplanted cancers of various kinds in mice and rats given guinea pig serum, horse serum, or rabbit serum. J Exp Med 98(6):565–582PubMedCentralCrossRefPubMedGoogle Scholar
  6. 6.
    Broome JD (1963) Evidence that the L-asparaginase of guinea pig serum is responsible for its antilymphoma effects. II. Lymphoma 6C3HED cells cultured in a medium devoid of L-asparagine lose their susceptibility to the effects of guinea pig serum in vivo. J Exp Med 118:121–148PubMedCentralCrossRefPubMedGoogle Scholar
  7. 7.
    Mashburn LT, Wriston JC Jr (1964) Tumor inhibitory effect of L-asparaginase from escherichia coli. Arch Biochem Biophys 105:450–452CrossRefPubMedGoogle Scholar
  8. 8.
    Jaffe N, Traggis D, Das L, Kim BS, Won H, Hann L, Moloney WC, Dohlwitz A (1972) Comparison of daily and twice-weekly schedule of L-asparaginase in childhood leukemia. Pediatrics 49(4):590–595PubMedGoogle Scholar
  9. 9.
    Tallal L, Tan C, Oettgen H, Wollner N, McCarthy M, Helson L, Burchenal J, Karnofsky D, Murphy ML (1970) E. coli L-asparaginase in the treatment of leukemia and solid tumors in 131 children. Cancer 25(2):306–320CrossRefPubMedGoogle Scholar
  10. 10.
    Ertel IJ, Nesbit ME, Hammond D, Weiner J, Sather H (1979) Effective dose of L-asparaginase for induction of remission in previously treated children with acute lymphocytic leukemia: a report from Childrens Cancer Study Group. Cancer Res 39(10):3893–3896PubMedGoogle Scholar
  11. 11.
    Haskell CM, Canellos GP (1969) l-asparaginase resistance in human leukemia–asparagine synthetase. Biochem Pharmacol 18(10):2578–2580CrossRefPubMedGoogle Scholar
  12. 12.
    Hutson RG, Kitoh T, Moraga Amador DA, Cosic S, Schuster SM, Kilberg MS (1997) Amino acid control of asparagine synthetase: relation to asparaginase resistance in human leukemia cells. Am J Physiol 272(5 Pt 1):C1691–C1699PubMedGoogle Scholar
  13. 13.
    Aslanian AM, Fletcher BS, Kilberg MS (2001) Asparagine synthetase expression alone is sufficient to induce l-asparaginase resistance in MOLT-4 human leukaemia cells. Biochem J 357(Pt 1):321–328PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Richards NG, Kilberg MS (2006) Asparagine synthetase chemotherapy. Annu Rev Biochem 75:629–654PubMedCentralCrossRefPubMedGoogle Scholar
  15. 15.
    Yong W, Zheng W, Zhang Y, Zhu J, Wei Y, Zhu D, Li J (2003) L-asparaginase-based regimen in the treatment of refractory midline nasal/nasal-type T/NK-cell lymphoma. Int J Hematol 78(2):163–167CrossRefPubMedGoogle Scholar
  16. 16.
    Yamaguchi M, Suzuki R, Kwong YL, Kim WS, Hasegawa Y, Izutsu K, Suzumiya J, Okamura T, Nakamura S, Kawa K, Oshimi K (2008) Phase I study of dexamethasone, methotrexate, ifosfamide, L-asparaginase, and etoposide (SMILE) chemotherapy for advanced-stage, relapsed or refractory extranodal natural killer (NK)/T-cell lymphoma and leukemia. Cancer Sci 99(5):1016–1020. doi: 10.1111/j.1349-7006.2008.00768.x CrossRefPubMedGoogle Scholar
  17. 17.
    Jaccard A, Petit B, Girault S, Suarez F, Gressin R, Zini JM, Coiteux V, Larroche C, Devidas A, Thieblemont C, Gaulard P, Marin B, Gachard N, Bordessoule D, Hermine O (2009) L-asparaginase-based treatment of 15 western patients with extranodal NK/T-cell lymphoma and leukemia and a review of the literature. Ann Oncol Off J Eur Soc Med Oncol ESMO 20(1):110–116. doi: 10.1093/annonc/mdn542 CrossRefGoogle Scholar
  18. 18.
    Molineux G (2003) Pegylation: engineering improved biopharmaceuticals for oncology. Pharmacotherapy 23(8 Pt 2):3S–8SCrossRefPubMedGoogle Scholar
  19. 19.
    Avramis VI, Sencer S, Periclou AP, Sather H, Bostrom BC, Cohen LJ, Ettinger AG, Ettinger LJ, Franklin J, Gaynon PS, Hilden JM, Lange B, Majlessipour F, Mathew P, Needle M, Neglia J, Reaman G, Holcenberg JS, Stork L (2002) A randomized comparison of native Escherichia coli asparaginase and polyethylene glycol conjugated asparaginase for treatment of children with newly diagnosed standard-risk acute lymphoblastic leukemia: a Children’s Cancer Group study. Blood 99(6):1986–1994CrossRefPubMedGoogle Scholar
  20. 20.
    Wang B, Relling MV, Storm MC, Woo MH, Ribeiro R, Pui CH, Hak LJ (2003) Evaluation of immunologic crossreaction of antiasparaginase antibodies in acute lymphoblastic leukemia (ALL) and lymphoma patients. Leukemia 17(8):1583–1588CrossRefPubMedGoogle Scholar
  21. 21.
    Shinnick SE, Browning ML, Koontz SE (2013) Managing hypersensitivity to asparaginase in pediatrics, adolescents, and young adults. J Pediatr Oncol Nurs Off J Assoc Pediatr Oncol Nurs 30(2):63–77. doi: 10.1177/1043454212471728 CrossRefGoogle Scholar
  22. 22.
    Panosyan EH, Seibel NL, Martin-Aragon S, Gaynon PS, Avramis IA, Sather H, Franklin J, Nachman J, Ettinger LJ, La M, Steinherz P, Cohen LJ, Siegel SE, Avramis VI (2004) Asparaginase antibody and asparaginase activity in children with higher-risk acute lymphoblastic leukemia: Children’s Cancer Group Study CCG-1961. J Pediatr Hematol Oncol 26(4):217–226CrossRefPubMedGoogle Scholar
  23. 23.
    Jaccard A, Gachard N, Marin B, Rogez S, Audrain M, Suarez F, Tilly H, Morschhauser F, Thieblemont C, Ysebaert L, Devidas A, Petit B, de Leval L, Gaulard P, Feuillard J, Bordessoule D, Hermine O (2011) Efficacy of L-asparaginase with methotrexate and dexamethasone (AspaMetDex regimen) in patients with refractory or relapsing extranodal NK/T-cell lymphoma, a phase 2 study. Blood 117(6):1834–1839. doi: 10.1182/blood-2010-09-307454 CrossRefPubMedGoogle Scholar
  24. 24.
    Rizzari C, Conter V, Stary J, Colombini A, Moericke A, Schrappe M (2013) Optimizing asparaginase therapy for acute lymphoblastic leukemia. Curr Opin Oncol 25(Suppl 1):S1–S9. doi: 10.1097/CCO.0b013e32835d7d85 CrossRefPubMedGoogle Scholar
  25. 25.
    van den Berg H (2011) Asparaginase revisited. Leuk Lymphoma 52(2):168–178. doi: 10.3109/10428194.2010.537796 CrossRefPubMedGoogle Scholar
  26. 26.
    Vrooman LM, Supko JG, Neuberg DS, Asselin BL, Athale UH, Clavell L, Kelly KM, Laverdiere C, Michon B, Schorin M, Cohen HJ, Sallan SE, Silverman LB (2010) Erwinia asparaginase after allergy to E. coli asparaginase in children with acute lymphoblastic leukemia. Pediatr Blood Cancer 54(2):199–205PubMedCentralPubMedGoogle Scholar
  27. 27.
    Roberts J, Holcenberg JS, Dolowy WC (1972) Isolation, crystallization, and properties of Achromobacteraceae glutaminase-asparaginase with antitumor activity. J Biol Chem 247(1):84–90PubMedGoogle Scholar
  28. 28.
    Wehner A, Harms E, Jennings MP, Beacham IR, Derst C, Bast P, Rohm KH (1992) Site-specific mutagenesis of Escherichia coli asparaginase II. None of the three histidine residues is required for catalysis. Eur J Biochem/FEBS 208(2):475–480CrossRefGoogle Scholar
  29. 29.
    Rajasekariah GH, Ryan JR, Hillier SR, Yi LP, Stiteler JM, Cui L, Smithyman AM, Martin SK (2001) Optimisation of an ELISA for the serodiagnosis of visceral leishmaniasis using in vitro derived promastigote antigens. J Immunol Methods 252(1–2):105–119CrossRefPubMedGoogle Scholar
  30. 30.
    Miller HK, Balis ME (1969) Glutaminase activity of L-asparagine amidohydrolase. Biochem Pharmacol 18(9):2225–2232CrossRefPubMedGoogle Scholar
  31. 31.
    Avramis VI, Panosyan EH (2005) Pharmacokinetic/pharmacodynamic relationships of asparaginase formulations: the past, the present and recommendations for the future. Clin Pharmacokinet 44(4):367–393CrossRefPubMedGoogle Scholar
  32. 32.
    Tardito S, Uggeri J, Bozzetti C, Bianchi MG, Rotoli BM, Franchi-Gazzola R, Gazzola GC, Gatti R, Bussolati O (2007) The inhibition of glutamine synthetase sensitizes human sarcoma cells to L-asparaginase. Cancer Chemother Pharmacol 60(5):751–758CrossRefPubMedGoogle Scholar
  33. 33.
    Fishburn CS (2007) The pharmacology of PEGylation: balancing PD with PK to generate novel therapeutics. J Pharm Sci 97(10):4167–4183CrossRefGoogle Scholar
  34. 34.
    Soares AL, Guimaraes GM, Polakiewicz B, de Moraes Pitombo RN, Abrahao-Neto J (2002) Effects of polyethylene glycol attachment on physicochemical and biological stability of E. coli L-asparaginase. Int J Pharm 237(1–2):163–170CrossRefPubMedGoogle Scholar
  35. 35.
    Fine BM, Kaspers GJ, Ho M, Loonen AH, Boxer LM (2005) A genome-wide view of the in vitro response to l-asparaginase in acute lymphoblastic leukemia. Cancer Res 65(1):291–299PubMedGoogle Scholar
  36. 36.
    Papageorgiou AC, Posypanova GA, Andersson CS, Sokolov NN, Krasotkina J (2008) Structural and functional insights into Erwinia carotovora L-asparaginase. Febs J 275(17):4306–4316CrossRefPubMedGoogle Scholar
  37. 37.
    Patel N, Krishnan S, Offman MN, Krol M, Moss CX, Leighton C, van Delft FW, Holland M, Liu J, Alexander S, Dempsey C, Ariffin H, Essink M, Eden TO, Watts C, Bates PA, Saha V (2009) A dyad of lymphoblastic lysosomal cysteine proteases degrades the antileukemic drug L-asparaginase. J Clin Invest 119(7):1964–1973. doi: 10.1172/JCI37977 PubMedCentralPubMedGoogle Scholar
  38. 38.
    Ollenschlager G, Roth E, Linkesch W, Jansen S, Simmel A, Modder B (1988) Asparaginase-induced derangements of glutamine metabolism: the pathogenetic basis for some drug-related side-effects. Eur J Clin Invest 18(5):512–516CrossRefPubMedGoogle Scholar
  39. 39.
    Durden DL, Salazar AM, Distasio JA (1983) Kinetic analysis of hepatotoxicity associated with antineoplastic asparaginases. Cancer Res 43(4):1602–1605PubMedGoogle Scholar
  40. 40.
    Durden DL, Distasio JA (1980) Comparison of the immunosuppressive effects of asparaginases from Escherichia coli and Vibrio succinogenes. Cancer Res 40(4):1125–1129PubMedGoogle Scholar
  41. 41.
    Storti E, Quaglino D (1970) Dysmetabolic and neurological complications in leukemia patients treated with L-asparaginase. Recent Results Cancer Res 33:344–349CrossRefPubMedGoogle Scholar
  42. 42.
    Distasio JA, Salazar AM, Nadji M, Durden DL (1982) Glutaminase-free asparaginase from vibrio succinogenes: an antilymphoma enzyme lacking hepatotoxicity. Int J Cancer 30(3):343–347CrossRefPubMedGoogle Scholar
  43. 43.
    Kumar S, Venkata Dasu V, Pakshirajan K (2011) Purification and characterization of glutaminase-free L-asparaginase from Pectobacterium carotovorum MTCC 1428. Bioresour Technol 102(2):2077–2082. doi: 10.1016/j.biortech.2010.07.114 CrossRefPubMedGoogle Scholar
  44. 44.
    Derst C, Henseling J, Rohm KH (2000) Engineering the substrate specificity of Escherichia coli asparaginase. II. Selective reduction of glutaminase activity by amino acid replacements at position 248. Protein Sci Publ Protein Soc 9(10):2009–2017. doi: 10.1110/ps.9.10.2009 CrossRefGoogle Scholar
  45. 45.
    Offman MN, Krol M, Patel N, Krishnan S, Liu J, Saha V, Bates PA (2011) Rational engineering of L-asparaginase reveals importance of dual activity for cancer cell toxicity. Blood 117(5):1614–1621. doi: 10.1182/blood-2010-07-298422 CrossRefPubMedGoogle Scholar
  46. 46.
    Riley V, Spackman D, Fitzmaurice MA, Roberts J, Holcenberg JS, Dolowy WC (1974) Therapeutic properties of a new glutaminase-asparaginase preparation and the influence of the lactate dehydrogenase-elevating virus. Cancer Res 34(2):429–438PubMedGoogle Scholar
  47. 47.
    Panosyan EH, Grigoryan RS, Avramis IA, Seibel NL, Gaynon PS, Siegel SE, Fingert HJ, Avramis VI (2004) Deamination of glutamine is a prerequisite for optimal asparagine deamination by asparaginases in vivo (CCG-1961). Anticancer Res 24(2C):1121–1125PubMedGoogle Scholar
  48. 48.
    Rotoli BM, Uggeri J, Dall’Asta V, Visigalli R, Barilli A, Gatti R, Orlandini G, Gazzola GC, Bussolati O (2005) Inhibition of glutamine synthetase triggers apoptosis in asparaginase-resistant cells. Cell Physiol Biochem 15(6):281–292CrossRefPubMedGoogle Scholar
  49. 49.
    Petronini PG, Urbani S, Alfieri R, Borghetti AF, Guidotti GG (1996) Cell susceptibility to apoptosis by glutamine deprivation and rescue: survival and apoptotic death in cultured lymphoma-leukemia cell lines. J Cell Physiol 169(1):175–185. doi: 10.1002/(SICI)1097-4652(199610)169:1<175::AID-JCP18>3.0.CO;2-C CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Wei-Wen Chien
    • 1
  • Soraya Allas
    • 2
  • Nicolas Rachinel
    • 1
  • Pierre Sahakian
    • 2
  • Michel Julien
    • 2
  • Céline Le Beux
    • 1
  • Claire-Emmanuelle Lacroix
    • 1
  • Thierry Abribat
    • 2
  • Gilles Salles
    • 1
    • 3
    Email author
  1. 1.Université Claude Bernard Lyon 1, UMR 5239, CNRS, ENS, HCLOullinsFrance
  2. 2.Alizé PharmaEcullyFrance
  3. 3.Hospices Civils de Lyon, Service d’HématologiePierre-BéniteFrance

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