The non specificity of specific nitric oxide synthase inhibitors

doi:10.1016/0006-291X(92)91266-S

Biochemical and Biophysical Research Communications

Volume 187, Issue 2, 16 September 1992, Pages 797-801

https://doi.org/10.1016/0006-291X(92)91266-S Get rights and content

Abstract

L-NAME (N_w-Nitro-L-arginine methylester) and L-NMMA (N^G-Monomethyl-L-arginine, monoacetate) are used widely as nitric oxide (NO) synthase inhibitors. Because of their functional groups (alcohols, amines and carboxylates), it appeared that they could interact with iron in a variety of systems. Using three in vitro models we observed these two compounds had inhibitory effects on cytochrome C reduction by ferrous iron, by ferrous iron accelerated by an unsaturated fatty acid or by epinephrine. This suggests that L-NAME and L-NMMA could have effects in iron containing systems found intracellularly apart from their inhibition of (NO) synthesis.

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Antitumour, Immunomodulatory activity and in silico studies of naphthopyranones targeting iNOS, a relevant target for the treatment of Helicobacter pylori infection
2018, Biomedicine and Pharmacotherapy
Citation Excerpt :
NO stimulates tumour growth and metastasis through increased vascularization; promotes the inactivation of the DNA repair proteins caused by the high affinity of NO for amino acids containing thiol residues [9]. Several substances are reported in the literature as being capable of inhibit its production [10–12], such as the non-selective inhibitor of nitric oxide synthase, Nω-Nitro-l-arginine methyl ester (L-NAME) [12], the selective inhibitors of the iNOS enzyme, N-(3 (1-yl) pyrimidin-4-yl]-carbamic acid tert-butyl ester (1400 W) and (N-[(1,3-benzodioxol-5-yl)methyl]-1-[2-(1H-imidazol-1-yl)pyrimidin-4-yl]-4-(methoxycarbonyl)-piperazine-2-acetamide) and natural products-inspired inhibitors such as the polyphenol butein, the biologically active component of Toxicodendron vernicifluum are all able to modulate NOS activity [13]. Because of the large number of secondary metabolites present in plants, their diversity and biological potential, its application in human health are diverse, becoming an important source for the drug development process [14,15].
The naphthopyranones paepalantine and 5-methoxy-3,4-dehydroxanthomegnin isolated from Paepalanthus sp, showed in previous studies antioxidant, anti-inflammatory, antitumour and antimicrobial potential, such as anti-Helicobacter pylori activity. H. pylori infection is one of the main causes of gastric cancer, causing an excessive inflammatory response through the neutrophils and macrophages infiltration, increasing the release of reactive species and thus inducing the production of pro-inflammatory mediators. In the present study, immunomodulatory activity of naphthopyranones in LPS-stimulated macrophages and cytotoxic action in gastric adenocarcinoma cell lines was evaluated. The potential of interaction of these substances in the iNOS binding site was investigated by molecular docking. Cytotoxic activity in gastric adenocarcinoma cells (AGS) was evaluated by the MTT assay. The results evidenced immunomodulatory potential by inhibiting the pro-inflammatory cytokines and nitric oxide produced by LPS-stimulated macrophages. Cytotoxic activity in AGS cell line was also reported. The results indicated that the studied naphthopyranones are viable alternatives in the treatment and prevention of H. pylori infection as well as the diseases related to this infection, especially gastric cancer.
Significance of nitric oxide synthases: Lessons from triple nitric oxide synthases null mice
2015, Journal of Pharmacological Sciences
Citation Excerpt :
The roles of NO derived from whole NOSs have been examined in pharmacological studies with non-selective NOSs inhibitors, such as Nω-nitro-L-arginine methyl ester (L-NAME) and NG-monomethyl-L-arginine (L-NMMA). However, the NOS inhibitors possess multiple non-specific actions, including antagonism of muscarinic acetylcholine receptors (13), generation of superoxide anions (14), inhibition of cytochrome c reduction (15), and inhibition of endothelium-independent relaxation induced by amiloride or cAMP (16). We also reported that vascular lesion formation caused by long-term treatment with L-NAME or L-NMMA is not mediated by the simple inhibition of eNOS in mice, and that activation of the tissue renin-angiotensin system and increased oxidative stress are involved in the long-term vascular effects of the L-arginine analogues in an NO-independent manner (17,18).
Nitric oxide (NO) is synthesized by three distinct NO synthases (neuronal, inducible, and endothelial NOSs), all of which are expressed in almost all tissues and organs in humans. The regulatory roles of NOSs in vivo have been investigated in pharmacological studies with non-selective NOS inhibitors. However, the specificity of the inhibitors continues to be an issue of debate, and the authentic significance of NOSs is still poorly understood. To address this issue, we generated mice in which all three NOS genes are completely disrupted. The triple NOSs null mice exhibited cardiovascular abnormalities, including hypertension, arteriosclerosis, myocardial infarction, cardiac hypertrophy, diastolic heart failure, and reduced EDHF responses, with a shorter survival. The triple NOSs null mice also displayed metabolic abnormalities, including metabolic syndrome and high-fat diet-induced severe dyslipidemia. Furthermore, the triple NOSs null mice showed renal abnormalities (nephrogenic diabetes insipidus and pathological renal remodeling), lung abnormalities (accelerated pulmonary fibrosis), and bone abnormalities (increased bone mineral density and bone turnover). These results provide evidence that NOSs play pivotal roles in the pathogenesis of a wide variety of disorders. This review summarizes the latest knowledge on the significance of NOSs in vivo, based on lessons learned from experiments with our triple mutant model.
Role of nitric oxide in optic nerve head blood flow regulation during an experimental increase in intraocular pressure in healthy humans
2013, Experimental Eye Research
Citation Excerpt :
In previous papers we have discussed this limitation in more detail (Petrig et al., 1999; Schmidl et al., 2013, 2012). In addition, some concerns have been raised dealing with the specificity of L-NMMA as an inhibitor of NO synthase (Iadecola et al., 1994; Peterson et al., 1992). We have, however, previously shown that the ocular hemodynamic effects of L-NMMA can be reversed by high-dose l-arginine (Schmetterer et al., 1997).
The present study set out to investigate whether nitric oxide, a potent vasodilator, is involved in the regulatory processes in optic nerve head blood flow during an experimental increase in intraocular pressure (IOP). The study was conducted in a randomized, double-masked, placebo-controlled, three way cross-over design. 12 healthy subjects were scheduled to receive either L-NMMA (an unspecific nitric oxide synthase inhibitor), phenylephrine (an α-adrenoceptor agonist) or placebo on three different study days. Optic nerve head blood flow was measured using laser Doppler flowmetry and IOP was increased stepwise with a suction cup. Mean arterial pressure (MAP) and IOP were measured non-invasively and ocular perfusion pressure (OPP) was calculated as OPP = 2/3 MAP-IOP. Administration of L-NMMA and phenylephrine significantly increased MAP and therefore OPP at rest (p < 0.01). L-NMMA significantly reduced baseline blood flow in the optic nerve head (p < 0.01). Application of the suction cup induced a significant increase in IOP and a decrease in OPP (p < 0.01). During the stepwise increase in IOP, some autoregulatory potential was observed until OPP decreased approximately −30% below baseline. None of the administered substances had an effect on this autoregulatory behavior (p = 0.49). The results of the present study confirm that the human optic nerve head shows some regulatory capacity during a decrease in OPP. Nitric oxide is involved in the regulation of basal vascular tone in the optic nerve head but does not seem to be involved in the regulatory mechanisms during an acute increase in IOP in young healthy subjects.
Vascular dilation by paeonol - A mechanism study
2010, Vascular Pharmacology
Citation Excerpt :
Buxton et al. has reported a blockage by L-NAME on muscarinic receptor which also existent in endothelial cell (Buxton et al., 1993). Inhibitory effect of L-NAME on cytochrome C reduction in vitro may also contribute to the decreased vasodilation by paeonol (Peterson et al., 1992). Furthermore, L-NAME may impair the urea cycle, in which L-arginine is a substrate for arginase (Brusilow and Horwich, 1989).
The goal of this study was to investigate the mechanism underlaying the vasodilatory effect of paeonol, a major active element from the root bark of Chinese herbs Paeonia suffruticosa Andr. and Cynanchum paniculatum (Bunge) Kitagawa. Paeonol relaxed isolated rat aorta rings by 95.6% while the 10⁻⁶ M forskolin-induced vasodilatation used as 100%. The EC₅₀ of vasodilatation by paeonol is 2.9 × 10⁻⁴ M. Although paeonol exerted endothelium-independent relaxation, L-NAME treatment inhibited paeonol-induced vasodilation of endothelium intact rings, while indomethacin did not. Both L-NAME and ODQ did not affect paeonol relaxation in the rings without endothelium. In addition, paeonol markedly elevated NO generation in cultured endothelial cells. Pre-treatment of propranolol, glibenclamide, TEA and BaCl₂ did not affect paeonol relaxation of endothelium removed rings. On the other hand, pre-treated of rings (without endothelium) with paeonol markedly blocked vasoconstriction induced by AngII, PGF_2α, 5-HT, dopamine, vasopressin, endothelin-1 and PE. The paeonol incubation also significantly attenuated KCl-induced contraction which mainly depended on Ca²⁺ influx. In Ca²⁺-free medium (containing 10⁻⁴ M of EGTA and 60 mM of KCl), paeonol suppressed the contraction curve of CaCl₂. In addition, paeonol also inhibited contraction by PE in Ca²⁺ free solution (containing 10⁻⁴ M of EGTA) which mainly relied on intracellular Ca²⁺ release. Whole-cell patch-clamp experiment showed that paeonol shifted the I–V curve and the peak value of calcium currents was significantly inhibited. In conclusion, our study suggested that voltage-dependent and receptor-operated Ca²⁺ channel, as well as intracellular Ca²⁺ release were all inhibited by paeonol. An intracellular Ca²⁺ regulatory mechanism may be responsible to potent vasodilatory effect of paeonol.
Spontaneous Myocardial Infarction and Nitric Oxide Synthase
2008, Trends in Cardiovascular Medicine
Citation Excerpt :
Direct activation of the renin-angiotensin system and increased oxidative stress, independent of endogenous NO inhibition, appear to be involved in the long-term vascular effects of those analogues (Suda et al. 2002, 2004). Consistent with our results, various nonspecific actions of the l-arginine analogues, including antagonism of muscarinic acetylcholine receptors (Buxton et al. 1993), generation of superoxide anions (Heim et al. 1991), inhibition of cytochrome c reduction (Peterson et al. 1992), and inhibition of endothelium-independent relaxation induced by amiloride or 3′-5′-cyclic adenosine monophosphate (Thomas and Ramwell 1991) have been reported. Male triply n/i/eNOS-/- mice showed markedly reduced survival as compared with male wild-type mice (Figure 1A).
Myocardial infarction (MI) is caused by coronary atherosclerosis and/or arteriosclerosis. Because endothelial nitric oxide synthase (eNOS) exerts powerful antiatherosclerotic/antiarteriosclerotic effects, it is speculated that blockade of eNOS activity might result in MI. However, neither genetic disruption of eNOS nor pharmacologic inhibition of eNOS activity induces MI in animals. On the other hand, intriguingly, genetic disruption of all three nitric oxide synthase (NOS) isoforms (neuronal NOS, inducible NOS, and eNOS) spontaneously caused MI accompanied by multiple cardiovascular risk factors of metabolic origin in mice. This is the first in vivo demonstration showing that the defective NOS system is involved in the pathogenesis of spontaneous MI. Based on the evidence, this review summarizes our current knowledge of spontaneous MI and NOS.
Skeletal Effects of Nitric Oxide: Novel Agent for Osteoporosis
2008, Principles of Bone Biology, Two-Volume Set
This chapter describes the skeletal effects of nitric oxide and its isoforms. The predominant nitric oxide synthases (NOS) isoform expressed by bone tissues is eNOS. iNOS is expressed in inflammatory and other pathological situations in osteoblasts, osteocytes, and in osteoclasts. eNOS is also expressed widely in vascular tissues within the bone, and consequent generation of lower levels of nitric oxide (NO) is likely to interact with bone cells. Synthesis of NO by these cells is stimulated by a variety of signals including hormones, cytokines, growth factors, fluid shear stress, and mechanical loading, resulting in a number of distinct physiological and pathophysiological responses. Nitric oxide donor compounds have been clinically used in humans for decades and are well tolerated; headaches are the only demonstrable adverse effects, which depend on the dose and the route of administration. Furthermore, NO has been shown to play a role in various physiological as well as pathological conditions. The most attractive novel indication for NO donor therapy is the prevention and treatment of osteoporosis in men and women. Recent research and developments have created the possibility of using NO donor compounds and designing new complex NO compounds capable of delivering NO directly into target tissues or to the bloodstream more efficaciously in a controlled or pulsatile manner. If a NO donor is proved to be effective in prevention of bone loss and, perhaps reduction of fracture risks, it may become a widely used, safe, and quite cost-effective therapeutic agent to combat osteoporosis.

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The non specificity of specific nitric oxide synthase inhibitors

Abstract

Biochem Biophys Res Comm

Eur J Pharm

FEBS Lett

J Biol Chem

FEBS Lett