Role of Enzymatic Activity in Producing an Antiproliferative Effect of L-Asparaginases

EXPERIMENTAL STUDIES , , ,

VS Pokrovskii1, MV Komarova2, SS Aleksandrova3, MV Pokrovskaya3, MS Kalish’yan1, SSh Karshieva1, EM Treshchalina1

1 N.N. Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478

2 S.P. Korolev Samara State Aerospace University, 34 Moskovskoye sh., Samara, Russian Federation, 443086

3 V.N. Orekhovich Scientific Research Institute of Biomedical Chemistry, 10 bld. 8 Pogodinskaya str., Moscow, Russian Federation, 119121

For correspondence: Vadim Sergeevich Pokrovskii, PhD, 24 Kashirskoye sh., Moscow, Russian Federation, 115478; Tel.: +7(499)324-14-09; e-mail: vadimpokrovsky@gmail.com

For citation: Pokrovskii VS, Komarova MV, Aleksandrova SS, et al. Role of Enzymatic Activity in Producing an Antiproliferative Effect of L-Asparaginases. Clinical oncohematology. 2015;8(2):120–8 (In Russ).

ABSTRACT

Background & Aims. E. coli (EcA) and Erwinia chrysanthemi (ErA) L-asparaginases have been used in antitumor chemotherapy for acute lymphoblastic leukemias since 1970s. Reports of their effect in a combined therapy of NK/T-cell and skin T-cell lymphomas have been published lately. The aim of this paper is to evaluate the relation between antiproliferative and enzymatic activities of L-asparaginases of different origin.

Methods. We conducted a prospective study of in vitro/in vivo enzymatic and antiproliferative activity of several new L-asparaginases: Yersinia pseudotuberculosis (YpA), Rhodospirillum rubrum (RrA), Wolinella succinogenes (WsA), Erwinia carotovora (EwA) in comparison with Escherichia coli L-asparaginase (EcA). Km, kcat and Vmax were calculated for evaluation of kinetic parameters. Cell lines of human transplantable tumors were used to analyze the cytotoxic activity, and DBA2 female mice with the body weight of 18–24 g with intraperitoneally transplanted Fisher lymphadenosis L5178Y (3–12 passage) from the bank of the N.N. Blokhin Russian Cancer Research Center were used for assessment of the antitumor activity.

Results. The use of type II L-asparaginases (EcA, EwA, YpA and WsA) permitted to determine a clear tendency: IC50 rises simultaneously with Km (r = 0.66; p = 0.007). Using single-dose range of 2000–8000 IU/kg in mice the highest efficacy was shown for L-asparaginases with Km = 0.017 and 0.054 mМ in comparison with L-asparaginases with lower enzymatic activity, as confirmed by corresponding odds ratio values in the stratified Cox proportional regression model. The pair-wise comparison of Kaplan-Meier curves and Cox regression method showed that WsA displays the best Km/antiproliferative activity ratio both in vitro and in vivo. In the L5178 mice model, it has been shown that WsA decreases the risk of treatment failure by 4–6 times, as compared to L-asparaginases with Km = 0.017 mМ (EcA and YpA) using a single-dose range of 500–1000 IU/kg and 2000–8000 IU/kg, RR = 0.16 and 0.28, respectively.

Conclusion. The obtained results confirmed the predictive value of enzymatic activity for demonstration of statistically significant antiproliferative activity both in vitro and in vivo. However, a statistical analysis demonstrated that this prognosis is not absolute for some enzymes or dose ranges. For instance, there might not be such correlation for high effective single dosed > 12 000 IU/kg.

Keywords: L-asparaginase, antitumor activity, enzymatic activity, mechanism of action.

Received: November 8, 2014

Accepted: January 28, 2015

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