Enzymatic oxidation of some non-phosphorylated derivatives of dihydronicotinamide

doi:10.1016/0006-291X(61)90272-8

Biochemical and Biophysical Research Communications

Volume 4, Issue 3, 10 March 1961, Pages 208-213

https://doi.org/10.1016/0006-291X(61)90272-8 Get rights and content

Abstract

This paper concerns the partial purification and properties of an enzyme that catalyzes the oxidation of reduced ribosyl nicotinamide (NRH) by vitamin K₃ (menadione) and certain other quinones. The enzyme is widely distributed in mammalian tissues. It is localized in the soluble portion of the cytoplasm. In the rat, the greatest activity of the enzyme was found in kidney, liver, heart and seminal vesicle. The purified enzyme also catalyzes the oxidation of certain N¹-alkyl-dihydronicotinamides, but it is completely inert toward phosphorylated derivatives of dihydronicotinamide such as the reduced forms of nicotinamide mononucleotide (NMNH), and of di- and triphosphopyridine nucleotides (DPNH and TPNH).

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The dawn of the 21st century will inevitably be confronted with numerous new medical challenges requiring the development of novel treatment agents and effective management strategies. In this article, we begin with an introduction that highlights and reviews the historical evidence leading to the classification of oxidoreductase quinone reductase 2 for the detoxification of quinones. The article then focuses on studies and approaches that result in advances and surprises regarding this enzyme. The subheadings that follow summarize novel, hitherto unreported observations pertaining to quinone reductase 2. We also discuss and provide perspectives on these findings.
Discovery of potent 4-aminoquinoline hydrazone inhibitors of NRH:quinoneoxidoreductase-2 (NQO2)
2019, European Journal of Medicinal Chemistry
Citation Excerpt :
NQO2 is a FAD-containing protein capable of oxidizing a variety of analogues of dihydronicotinamide. It is a structural analogue of NAD(P)H:quinone oxidoreductase 1 (NQO1, DT-diaphorase, EC.1.6.99.2) and while the two enzymes have similar properties there are major functional differences [10–12]. In particular, the two enzymes catalyse the reduction of various quinones into hydroquinones [12–15].
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Cytotoxicity and NQO2–inhibitory activity in vitro was evaluated using ovarian cancer SKOV-3 and TOV-112 cells (expressing high and low levels of NQO2, respectively). Generally, the hydrazones were more toxic than hydrazide analogues and further, toxicity is unrelated to cellular NQO2 activity. Pharmacological inhibition of NQO2 in cells was measured using the toxicity of CB1954 as a surrogate end-point. Both the hydrazone and hydrazide derivatives are functionally active as inhibitors of NQO2 in the cells, but at different inhibitory potency levels. In particular, 4-((2-(6-methoxy-2-methylquinolin-4-yl)hydrazono)methyl)phenol has the greatest potency of any compound yet evaluated (53 nM), which is 50-fold lower than its toxicity IC₅₀. This compound and some of its analogues could serve as useful pharmacological probes to determine the functional role of NQO2 in cancer development and response to therapy.
Evaluation of analogues of furan-amidines as inhibitors of NQO2
2018, Bioorganic and Medicinal Chemistry Letters
Inhibitors of the enzyme NQO2 (NRH: quinone oxidoreductase 2) are of potential use in cancer chemotherapy and malaria. We have previously reported that non-symmetrical furan amidines are potent inhibitors of NQO2 and here novel analogues are evaluated. The furan ring has been changed to other heterocycles (imidazole, N-methylimidazole, oxazole, thiophene) and the amidine group has been replaced with imidate, reversed amidine, N-arylamide and amidoxime to probe NQO2 activity, improve solubility and decrease basicity of the lead furan amidine. All compounds were fully characterised spectroscopically and the structure of the unexpected product N-hydroxy-4-(5-methyl-4-phenylfuran-2-yl)benzamidine was established by X-ray crystallography. The analogues were evaluated for inhibition of NQO2, which showed lower activity than the lead furan amidine. The observed structure-activity relationship for the furan-amidine series with NQO2 was rationalized by preliminary molecular docking and binding mode analysis. In addition, the oxazole-amidine analogue inhibited the growth of Plasmodium falciparum with an IC₅₀ value of 0.3 μM.
Non-symmetrical furan-amidines as novel leads for the treatment of cancer and malaria
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NRH:quinone oxidoreductase 2 enzyme (NQO2) is a potential therapeutic target in cancer and neurodegenerative diseases, with roles in either chemoprevention or chemotherapy. Here we report the design, synthesis and evaluation of non-symmetrical furan-amidines and their analogues as novel selective NQO2 inhibitors with reduced adverse off-target effects, such as binding to DNA. A pathway for the synthesis of the non-symmetrical furan-amidines was established from the corresponding 1,4-diketones. The synthesized non-symmetrical furan-amidines and their analogues showed potent NQO2 inhibition activity with nano-molar IC₅₀ values. The most active compounds were non-symmetrical furan-amidines with meta- and para-nitro substitution on the aromatic ring, with IC₅₀ values of 15 nM. In contrast to the symmetric furan-amidines, which showed potent intercalation in the minor grooves of DNA, the synthesized non-symmetrical furan-amidines showed no affinity towards DNA, as demonstrated by DNA melting temperature experiments. In addition, Plasmodium parasites, which possess their own quinone oxidoreductase PfNDH2, were inhibited by the non-symmetrical furan-amidines, the most active possessing a para-fluoro substituent (IC₅₀ 9.6 nM). The high NQO2 inhibition activity and nanomolar antimalarial effect of some of these analogues suggest the lead compounds are worthy of further development and optimization as potential drugs for novel anti-cancer and antimalarial strategies.
Characterization of cofactors, substrates and inhibitor binding to flavoenzyme quinone reductase 2 by automated supramolecular nano-electrospray ionization mass spectrometry
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Citation Excerpt :
QR1 and QR2 are closely related [2], since they share 59% (cDNA) and 44% (protein) similarity in their sequences, with the interesting difference that QR1 has a 43 amino acid longer C-terminus than QR2 [3]. Due to this particular fact, QR2 does not recognize NADH and NADPH as co-substrates but rather natural compounds possibly derived from those, such as N-ribosyl- or N-methyl-dihydronicotinamide [4,5]. Secondly, two papers are absolutely crucial in understanding the role of QR1 and QR2 in animals.
Quinone reductase 2 (QR2) is a cytosolic homodimeric enzyme implicated in the reduction of quinone in the presence of natural derivatives of NADH such as N-ribosyldihydronicotinamide. QR2 does not recognize NADH or NADPH as co-substrates, unlike quinone reductase 1 (QR1). This feature is not the only unusual one of this enzyme. Although it resembles quinone reductase 1, the well-described detoxifying enzyme, QR2 does not share many features with QR1. Particularly, it does not seem to have a similar detoxifying function in cells. Therefore, starting from basic knowledge on QR2 and adding up to previous works published on the enzyme, we wanted to rebuild its biochemical description because some of the recently described characteristics are surprising, and merit further explorations. For example, QR2 seems to be over-expressed in neurodegenerative diseases, and this over-expression seems to be linked to a worsening of the pathological conditions. Indeed, our specific inhibitors of QR2, tested in vivo, show outstanding properties impairing memory loss. These observations led us to further describe, at the molecular level, the relationship between QR2 and some of its inhibitors and co-substrates. In the present paper, we address this question using non-denaturing supramolecular nano-electrospray ionization mass spectrometry. This characterization helps understand the physical relationship between inhibitors such as resveratrol or melatonin and the enzyme.
In silico identification and biochemical evaluation of novel inhibitors of NRH:quinone oxidoreductase 2 (NQO2)
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Enzymatic oxidation of some non-phosphorylated derivatives of dihydronicotinamide

Abstract

Biochem. Biophys. Res. Comm

Biochem. Biophys. Res. Comm

J. Biol. Chem

J. Biol. Chem

J. Biol. Chem