Enzymic degradation of plasma arginine using arginine deiminase inhibits nitric oxide production and protects mice from the lethal effects of tumour necrosis factor α and endotoxin

Abstract

Septic shock is mediated in part by nitric oxide (NO) and tumour necrosis factor α (TNFα). NO is synthesized primarily from extracellular arginine. We tested the ability of an arginine-degrading enzyme to inhibit NO production in mice and to protect mice from the hypotension and lethality that occur after the administration of TNFα or endotoxin. Treatment of BALB/c mice with arginine deiminase (ADI) formulated with succinimidyl succinimide polyethylene glycol of Mr 20000 (ADI-SS PEG20000) eliminated all measurable plasma arginine (from normal levels of ∼155μM arginine to 2μM). In addition, ADI-SS PEG20000 also inhibited the production of NO, as quantified by plasma nitrate+nitrite. Treatment of mice with TNFα or endotoxin resulted in a dose-dependent increase in NO production and lethality. Pretreatment of mice with ADI-SS PEG20000 resulted in increased resistance to the lethal effects of TNFα and endotoxin. These observations are consistent with NO production resulting, to some extent, from the metabolism of extracellular arginine. The toxic effects of TNFα and endotoxin may be partially inhibited by enzymic degradation of plasma arginine by ADI-SS PEG20000. Interestingly, pretreatment with ADI-SS PEG20000 did not inhibit the anti-tumour activity of TNFα in vitro or in vivo. This treatment may allow greater amounts of TNFα, as well as other cytokines, to be administered while abrogating side effects such as hypotension and death.

Renal cell carcinoma does not express argininosuccinate synthetase and is highly sensitive to arginine deprivationviaarginine deiminase

“…These results suggest that ADI treatment did not directly suppress the expression of VEGF, but clearly reduced angiogenic activity in tumor tissue. ADI inhibited the production of nitric oxide (NO) by nitric oxide synthase (NOS) through the modulation of extracellular level of arginine,5, 32, 33 and NO plays a role in angiogenesis because it could modulate the activity of vascular endothelial growth factor, basic fibroblast growth factor and matrix metalloproteinases 34, 35. Arginine deprivation by ADI could also alter the levels of polyamines, proline and glutamate.…”

Purification, immobilization, and biochemical characterization ofl‐arginine deiminase from thermophilicAspergillus fumigatusKJ434941: Anticancer activityin vitro

“…ADI from Mycoplasma arginini has a powerful anticancer potency toward various tumors . The dependency of tumor cells on blood l ‐arginine for rapid proliferation was reported more than 50 years ago.…”

“…ADI was well characterized from Bacillus cereus , Pseudomonas sp, Leuconostoc oenos , Lactobacillus sp, Listeria monocytogenes , Clostridium perfringens , and Mycoplasma sp . ADI was absolutely absent in mammals . However, few studies on fungal ADI have been reported, despite its hypothesized affordable biochemical properties compared with bacterial ADI …”

“…Regarding prior clinical trials, ADI from Mycoplasma arginini displayed a promising anticancer activity toward various types of tumor cells . However, the main challenges of M. arginini ADI were the higher antigenicity, short half‐life time in vivo and remarkable lower thermal stability . Recently, PEGylation of M. arginini ADI could greatly reduce its antigenicity, increase its half‐life time, and improves its pharmacokinetic properties .…”

Recombinant arginine deiminase reduces inducible nitric oxide synthase iNOS‐mediated neurotoxicity in a coculture of neurons and microglia

“…Recombinant arginine deiminase (rADI) with the modification of polyethylene glycol was recently developed as an antitumor drug for patients with hepatocellular carcinoma and melanoma (Shen and Shen, 2006). In addition, rADI was shown to have beneficial effects on the maintenance of blood pressure in septic animals because of suppression of iNOS‐induced NO production (Thomas et al, 2002). However, the effects of rADI on NO‐induced neuronal diseases remain unknown.…”