Diazeniumdiolates:: Pro- and antioxidant applications of the “NONOates”

doi:10.1016/S0891-5849(00)00251-3

Free Radical Biology and Medicine

Volume 28, Issue 10, 15 May 2000, Pages 1463-1469

https://doi.org/10.1016/S0891-5849(00)00251-3 Get rights and content

Abstract

Diazeniumdiolates are compounds containing the X-[N(O)NO]⁻ structural unit that as a class offer many advantages as tools for probing the roles of nitric oxide (NO) in biological redox processes. Available examples in which X is a secondary amine group spontaneously generate up to two molecules of NO per [N(O)NO]⁻ unit when dissolved in aqueous media; their half-lives range from 2 s (for X = L-prolyl) to 20 h [for X = (H₂NCH₂CH₂)₂N] at pH 7.4 and 37°C, and are in general relatively little influenced by medium effects or metabolism. When X = O⁻ (Angeli’s salt), first-order dissociation produces NO⁻ rather than NO, but the ion becomes an NO source on 1-electron oxidation; diazeniumdiolate-derived NO can also be used to generate reactive nitrogen/oxygen species with higher nitrogen oxidation states (+3 and +4) in the presence of selected oxidizing agents. The advantages of diazeniumdiolates in biomedical research are briefly illustrated with examples from the recent literature probing NO’s role in inhibiting oxidative drug metabolism, radical-induced lipid oxidation, the cytotoxicity of reactive oxygen species, and ischemia-induced vascular reoxygenation injury. Future work with this compound class should provide further insight into the mechanisms of NO’s involvement in pro- and antioxidant processes, and may well lead to important medicinal advances, including reversal of cerebral vasospasm and radiosensitization of hypoxic tumors.

Introduction

A diazeniumdiolate is any chemical species containing the [N(O)NO]⁻ functional group [1]. Compounds bearing this structural unit are seeing increasing use both as biomedical research tools and as lead compounds for potential clinical applications [2]. A primary basis for their utility is that many of them decompose spontaneously in aqueous media to release the critical bioregulatory species, nitric oxide (NO), as in Eqn. 1. The structures of some useful examples that are commercially available are shown in Fig. 1 , along with their 37°C/pH 7.4 half-lives and the identifying acronyms we recommend for each as a means of standardizing their nomenclature.¹ $X^{+} N ⧸ O^{−} NO^{−} pH 7.4 → X^{−} +2 NO$

Section snippets

Physicochemical characteristics of the diazeniumdiolates: advantages they offer the biomedical researcher

With many other classes of NO-generating compounds currently available [3], one might well ask what advantages diazeniumdiolates have as biochemical research reagents over the S-nitrosothiols (several examples of which are found in human physiological fluids) or such life-saving clinical agents as the nitrate esters and the metal nitrosyl compound sodium nitroprusside. While we would in no way want to disparage these and other important sources of bioactive NO, we believe the diazeniumdiolates

Illustrative research applications

We can illustrate what we believe to be the significant advantages of this compound class by citing some artful studies from the recent literature of NO’s role in biological oxidation-reduction phenomena.

Possible clinical applications

The examples cited above suggest that diazeniumdiolates designed to generate molecular NO at the right time, rate, and bodily location might indeed help to combat oxidative stress, for example, on reperfusing an ischemic vessel or vascular bed. In fact, two different groups have exploited the kinetic reliability of the diazeniumdiolates to inhibit reperfusion injury and shorten or block ischemic episodes by selectively dilating spastic vessels before or soon after they begin to constrict. Pluta

Conclusion

In our view, the diazeniumdiolates constitute the NO donors of choice for a variety of biomedical applications, especially those requiring known, controllable rates of NO release in the physiological milieu. Thus one can depend on DETA/NO (Fig. 1) to provide a nearly constant rate of NO generation in cell culture media, one that decreases by only about 50% during a 24 h day at 37°C in a pH 7.4 medium. At the other kinetic extreme, a very fast-acting diazeniumdiolate like PROLI/NO (Fig. 1) can

Acknowledgements

Supported in part by the National Cancer Institute under contract No. NO1- CO-56000.

References (33)

L.K. Keefer et al.
“NONOates” (1-substituted diazen-1-ium-1,2-diolates) as nitric oxide donorsConvenient nitric oxide dosage forms
Methods Enzymol.
(1996)
D.A. Wink et al.
Inhibition of cytochromes P450 by nitric oxide and a nitric oxide-releasing agent
Arch. Biochem. Biophys.
(1993)
J. Kanner et al.
Nitric oxide as an antioxidant
Arch. Biochem. Biophys.
(1991)
S.P.A. Goss et al.
The effect of nitric oxide release rates on the oxidation of human low density lipoprotein
J. Biol. Chem.
(1997)
D.A. Wink et al.
The effect of various nitric oxide-donor agents on hydrogen peroxide-mediated toxicitya direct correlation between nitric oxide formation and protection
Arch. Biochem. Biophys.
(1996)
D.A. Wink et al.
Nitric oxide protects against alkyl peroxide-mediated cytotoxicityfurther insights into the role nitric oxide plays in oxidative stress
Arch. Biochem. Biophys.
(1995)
G. Raspi et al.
Oxidative determination of α-hyponitrate with alkaline hexacyanoferrate(III)
Anal. Chim. Acta
(1974)
J.M. Fukuto et al.
Conversion of nitroxyl (HNO) to nitric oxide (NO) in biological systemsthe role of physiological oxidants and relevance to the biological activity of HNO
Biochem. Biophys. Res. Commun.
(1993)
A.M. Miles et al.
Modulation of superoxide-dependent oxidation and hydroxylation reactions by nitric oxide
J. Biol. Chem.
(1996)
T. Okamoto et al.
Activation of human neutrophil procollagenase by nitrogen dioxide and peroxynitritea novel mechanism for procollagenase activation involving nitric oxide
Arch. Biochem. Biophys.
(1997)

A.J. Gow et al.

Electrochemical detection of nitric oxide in biological systems

Microchem. J.

(1997)

L.K. Keefer

Nitric oxide-releasing compoundsfrom basic research to promising drugs

CHEMTECH

(1998)

Y.-C. Hou et al.

Current trends in the development of nitric oxide donors

Curr. Pharm. Des.

(1999)

E.A. Kowaluk et al.

Spontaneous liberation of nitric oxide cannot account for in vitro vascular relaxation by S-nitrosothiols

J. Pharmacol. Exp. Ther.

(1990)

McAninly, J.; Williams, D. L. H.; Askew, S. C.; Butler, A. R.; Russell, C. Metal ion catalysis in nitrosothiol (RSNO)...

J.S. Beckman

Oxidative damage and tyrosine nitration from peroxynitrite

Chem. Res. Toxicol.

(1996)

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Citation Excerpt :
To gain possible insight into the mechanism of NO regulation, we examined the effects of other NO donors on CYP2B6V5 down-regulation. Spermine NONOate, like DPTA and NOC18 is a pure NO donor [28,29], releasing NO radicals in a time dependent manner. The half-life of spermine NONOate is 37 min.
We showed previously that rat cytochrome P450 CYP2B1 undergoes NO-dependent proteasomal degradation in response to inflammatory stimuli, and that the related human enzyme CYP2B6 is also down-regulated by NO in primary human hepatocytes. To investigate the mechanism of CYP2B6 down-regulation, we made several cell lines (HeLa and HuH7 cells) in which native CYP2B6 or CYP2B6 with a C-terminal V5 tag (CYP2B6V5) are expressed from a lentiviral vector with a cytomegalovirus promoter. Native CYP2B6 protein was rapidly down-regulated in HeLa cells within 3 h of treatment with the NO donor (Z)−1-[2-(2-Aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, while its mRNA level was not down-regulated. Treatment of the cells with the NO donor (Z)−1-[N-(3-aminopropyl)-N-(3-ammoniopropyl)amino]diazen-1-ium-1,2-diolate also resulted in rapid down-regulation of CYP2B6 activity, measured as the formation of 7-hydroxy-4-trifluoromethylcoumarin, as well as 2B6 protein in the CYP2B6 HeLa cell line. CYP2B6V5 was also down-regulated by NO donors in HuH7 cells. Down-regulation was observed in the presence of cycloheximide, demonstrating that this occurs via a post-translational mechanism. We generated a HeLa cell line expressing both CYP2B6V5 and human nitric oxide synthase 2 (NOS2), the latter under positive control by tetracycline. The cellular NO produced by doxycycline treatment also effectively down-regulated CYP2B6 protein, which was blocked by the co-treatment with the NOS2 competitive inhibitor L-N^G-nitroarginine methyl ester (L-NAME). We next investigated the proteolytic enzymes responsible for NO-dependent CYP2B6 degradation. Neither calpain inhibitors (N-Acetyl-L-leucyl-L-leucyl-L-norleucinal, carbobenzoxy-valinyl-phenylalaninal), nor lysosomal protease inhibitors (3-methyladenine and chloroquine) inhibited the NO dependent CYP2B6V5 down-regulation. The proteasome inhibitors MG132 and bortezomib attenuated, but did not completely block the NO-induced down-regulation in the HuH7 cell line. However, when cells were co-treated with NO donor and proteasome inhibitors, high molecular mass species could be detected on native CYP2B6 as well as CYP2B6V5 Western blots. Further investigation demonstrated that CYP2B6 protein was polyubiquitinated and this was dramatically enhanced by co-treatment with NO donor and bortezomib. Taken together, our data demonstrate that CYP2B6 is down-regulated in an NO-dependent manner via ubiquitination and proteasomal degradation.
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A novel class of O²-(2,4-dinitrophenyl)-1-[N,N-bis(2-substituted ethyl)amino]diazen-1-ium-1,2-diolates 4–6 were designed, synthesized, and biologically evaluated. The most active compound 6 caused significant DNA damage by releasing N,N-bis(2-TsO ethyl)amine and two molecules of nitric oxide (NO) after activation by GST/GSH in cancer cells, being more cytotoxic against three cancer cell lines than a well-known diazeniumdiolate-based anticancer agent JS-K, suggesting that the strategy has potential to extend to other O²-derived diazeniumdiolates to improve anticancer activity.
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¹: We recommend that this class of compounds be referred to henceforth as diazeniumdiolates, rather than as “NONOates”, “Drago complexes”, “NOC” compounds, etc., because the “diazeniumdiolates” designation is derived from International Union of Pure and Applied Chemistry nomenclature rules and thus reflects the structure of the functional group in a universally recognizable way [1]. The acronyms for the individual compounds in this series (including those used in Fig. 1) are similarly derived from structural considerations [2]. Our aim in making these recommendations is to clear up what we perceive to be some unfortunate confusion in the literature that has arisen from the use of alternate nomenclature systems devoid of structural significance.

²: Anthony Fitzhugh received his bachelor’s degree in chemistry from Princeton in 1977 and his M.D. from George Washington University in 1982. His research at SAIC Frederick, where he is now a Senior Scientist, and previously at Biocipher BF, has historically focused on medicinal chemistry, with emphasis on antimicrobial and anticancer agents, particularly folate analogues. After Larry Keefer received his B.A. and Ph.D. degrees from Oberlin College (1961, chemistry) and the University of New Hampshire (1965, organic chemistry), respectively, he held research positions at the Chicago Medical School and the University of Nebraska College of Medicine before coming to the National Cancer Institute (NCI) in 1971. He and his colleagues in the NCI’s Chemistry Section, Laboratory of Comparative Carcinogenesis, are intensively studying the chemistry and pharmacology of the diazeniumdiolates as nitric oxide sources for biomedical applications.

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Forum: therapeutic applications of reactive oxygen and nitrogen species in human diseaseDiazeniumdiolates:: Pro- and antioxidant applications of the “NONOates”1

Abstract

Introduction

Section snippets

Physicochemical characteristics of the diazeniumdiolates: advantages they offer the biomedical researcher

Illustrative research applications

Possible clinical applications

Conclusion

Acknowledgements

Methods Enzymol.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Anal. Chim. Acta

Biochem. Biophys. Res. Commun.

J. Biol. Chem.

Arch. Biochem. Biophys.

Microchem. J.

Nitric oxide-releasing compoundsfrom basic research to promising drugs

CHEMTECH

Current trends in the development of nitric oxide donors

Curr. Pharm. Des.

Spontaneous liberation of nitric oxide cannot account for in vitro vascular relaxation by S-nitrosothiols

J. Pharmacol. Exp. Ther.

Oxidative damage and tyrosine nitration from peroxynitrite

Chem. Res. Toxicol.

Forum: therapeutic applications of reactive oxygen and nitrogen species in human disease
Diazeniumdiolates:: Pro- and antioxidant applications of the “NONOates”¹