Nitric oxide (NO) is an endogenic product from plants, bacteria, and animal cells that has many important effects in those organisms. It is produced by nitric oxide synthase (NOS), which is found in main three isoforms, namely endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS). It has an important role in homeostasis in different physiological systems, such as micro- and macro-vascularization, inhibition of platelet aggregation, and neurotransmission regulation in the central nervous, gastrointestinal, respiratory, and genitourinary systems. However, its overproduction has been associated with diseases such as arthritis, asthma, cerebral ischemia, Parkinson’s disease, neurodegeneration, and seizures. For this reason, and due to better understanding of the molecular mechanisms by which NO provokes those diseases, the interest on the design of NOS inhibitors with therapeutic purposes has highly increased. Based on the foregoing considerations, the proposal of this chapter is to show an overview about the design strategies, mechanism of action at the molecular level, and the main advances toward the search for selective NOS inhibitors available in the literature.
- nitric oxide synthase isoforms
- structure-based drug design
- enzymatic inhibition
- heterocyclic compounds
Nitric oxide (NO) is a diatomic neutral molecule, produced by bacteria, plants, and animals. Having one unpaired electron, its effect in biological system is related to the stabilization of this electron. It acts as dissolved nonelectrolyte in the organisms, except for the lungs, where it is found in gaseous state [1–3].
NO has gain importance mainly in the 1990s, and from then on, it has been studied to obtain interesting pharmacological effects. A review by Serafim and collaborators describes the state of the art of this compound use in drug design .
The basal NO production has an important contribution to homeostasis in different physiological systems, such as micro- and macro-vascularization, inhibition of platelet aggregation, and neurotransmission regulation in central nervous, gastrointestinal, respiratory, and genitourinary systems. However, NO overproduction has been strongly associated with some diseases such as arthritis, asthma, cerebral ischemia, Parkinson’s disease, neurodegeneration, and seizures [5–9].
Nitric oxide synthase (NOS) is the enzyme responsible for NO biosynthesis, and there are three main kinds of NOS isoforms [endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS)]. They are tetramers, constituted of two NOS monomers associated with two calmodulin monomers (CaM), and contain relatively tightly bound cofactors, BH4, FAD, FMN, and iron protoporphyrin IX (heme group). Their chemical function is to catalyze the reaction of l-arginine, NADPH, and oxygen to synthesize free radical NO, l-citrulline, and NADP (Figure 1) .
The substrate l-arginine establishes H-bond networks inside the catalytic site of NOS isoforms with the heme group, mainly due to the guanidine group, which is crucial to bind tightly using a salt-bridge interaction with the conserved carboxylate of Glu597 in human nNOS, Glu377 for iNOS and Glu361 for eNOS. In addition, l-arginine establishes H-bonds with the amide carbonyl from Trp592, in nNOS; with Trp372, in iNOS; and with Trp356, in eNOS. Moreover, α-amino group of l-arginine interacts through H-bond with the heme propionate side chain, and the guanidine
Exacerbated induction of iNOS is associated with septic shock, inflammatory, and noninflammatory impairment processes in different tissues/organs, and, likewise, the nNOS is triggered in neurotoxicity, neurodegeneration process, and proliferation increase of some neoplastic cell lines. Depending on the clinical condition, decreasing NO levels is necessary, and excellent benefits might be achieved using NOS inhibitors. However, it is much important not to inhibit eNOS, because of its central role in smooth muscle relax, controlling vascular tone and blood pressure [12–14].
The first inhibitors designed (during the1980s and early 1990s) were based on l-arginine, the substrate of the enzyme, and this approach led to potent compounds but with poor selectivity level among the isoforms. In the late 1990s, the first crystal structure of iNOS and eNOS was unveiled, showing the high degree of similarity particularly in the active site of both isoforms. The nNOS crystal structure was reported in 2002, allowing the design of selective inhibitors [15, 16]. It is worth noting that changes in some amino acids of the isoforms lead to differences in electronic and steric effects on the binding site region, which can be interesting for designing selective inhibitors [11, 15]. The active collaboration between Richard Silverman and Thomas Poulos’ groups has significantly contributed to this field, and some of their papers are discussed in this chapter.
NOS isoforms were validated as target for new drugs soon after their X-ray crystallography was available. From then on the design of effective and selective inhibitors has been an important approach in modern drug discovery involving NO biochemical pathways related to many dysfunctions of the human organism [12, 17–19].
2. Experimental studies
2.1. Inducible nitric oxide synthase (iNOS) inhibitors
Garvey and collaborators (1994) were the first to report highly selective iNOS inhibitors. The compounds were isothiourea derivatives (Figure 3—
The selective iNOS inhibition by aminoguanidine (Figure 3—
In 2000, Hagmann and collaborators explored the structure-activity relationships of a series of substituted 2-aminopyridines. Compounds 4,6-dialkyl substituted (Figure 3—
On the other hand, the 1,2-dihydro-4-quinazolinamine compound
Structural-based approach using crystal structure and mutagenesis have identified specific induced-fit binding mode, which can generate some conformational changes toward a new specific cavity. Garcin and coworkers showed the
Other compounds, as acetamidine derivatives (Figure 5—
A protein called SPSB2 plays an important role in modulating the activity of iNOS through its proteasomal degradation in defense cells. Since this complex is blocked, the NO production from iNOS is prolonged, increasing the killing activity against pathogen microorganisms, making it an interesting anti-infective target . Some cyclic peptidomimetic compounds were designed using this strategy, and the most potent compound
High-throughput screening (HTS) strategy has been used to identify new iNOS inhibitors hits such as the compound
Phenylpyrroles, pyrazoles, urea kynurenamines, ethynylcyanodienones, and amidine derivatives (Figure 6—
Natural products have been a rich source of new bioactive molecules. Some examples in the NOS inhibition are a sesquiterpenoid, isolated from
2.2. Neuronal nitric oxide synthase (nNOS) inhibitors
In the beginning of the 1990s, efforts to design selective nNOS inhibitor compounds were addressed, using the substrate l-arginine as the prototype molecule. Series of analogs was synthesized to evaluate which molecular change could interfere in the ligand activity and selectivity over other isoforms. The first selective compound over nNOS was l-nitroarginine (Figure 7), producing hypertension in animals due to the lack of selectivity over eNOS. In addition, many peptide analogs were synthesized trying to obtain more promising compounds. After the X-ray crystal complex elucidation, structure-activity relationship findings of several scaffolds have been explored to identify the molecular basis of improving the selectivity toward neuronal isoform [15, 19].
The non-arginine-based compound 7-nitroindazole (Figure 7—
Entrena and collaborators, by using kynurenamine scaffold (Figure 7—
Aminopyridine is an attractive pharmacophoric group to bind in different regions of nNOS through H-bond. Using this moiety, compound
Other interesting structures such as 2-aminoquinolines are effective scaffold to be included in the structure of nNOS inhibitors. Crystallography studies showed that those compounds act as competitive arginine mimics. This scaffold makes important H-bonds with the active-site Glu residue, and the non-coordinating aryl rings are stabilized in a hydrophobic pocket in the extremity of the substrate access channel. Moreover, this structural class showed good pharmacokinetic properties (Figure 8—
Exploring the heme-coordinating potential of imidazole group, a series of 2,4-disubtituted pyrimidine compounds (Figure 8—
Studies using aminopyridine-based scaffold with pyridine linker (Figure 8—
Rational strategy for identifying new nNOS inhibitors using a combination of virtual screening approach based on 3D pharmacophore model and molecular docking was able to identify a hit compound structurally different from the available inhibitors (Figure 8—
2.2.1. Double-headed nNOS inhibitors
Double-headed compounds have been explored by researches with the aim of obtaining high affinity binding in nNOS. Attaching a double-headed aminopyridine moiety in a compound led to a very potent (
Other symmetric double-headed aminopyridine series without charge groups were designed to contain a tail on the central aromatic ring. The objective is to achieve an interaction in the electronegative region in the catalytic site, since only the neuronal isoform has Asp597 in this region. Derivative
Furthermore, double-headed inhibitors containing chiral linkers derived from natural amino acids were designed and synthesized. The best compound (Figure 10—
2-Amino-4-methylpyridine groups with a chiral linker derived from proline were designed as selective nNOS inhibitors. They showed to be interesting as they can interact in a unique orientation, what led to selectivity toward neuronal isoform. The aminopyridine groups interact with a Glu592 residue and the heme propionate in nNOS active site. In addition, the nitrogen from pyrrolidine linker is important to contribute to additional hydrogen bonds to the heme propionate, resulting in the most potent compound (
In addition, using chiral double-headed inhibitors, the α-amino-functionalized aminopyridine derivative
The non-chiral double-headed thiophene-2-carboximidamide compound (Figure 10—
2.3. Bacterial nitric oxide synthase (bNOS) inhibitors
Bacterial nitric oxide synthase (bNOS) is present in many Gram-positive microorganisms and has been described as part of their defense system against other species and the oxidative stress provoked by antibiotics through NO releasing. Therefore, bNOS inhibition can increase the antibiotic potential and be harmful to bacterial cell .
A screening showed that some known nNOS inhibitors can decrease significantly the percent survival of
Exploring the potential of bNOS as a drug target, high selectivity levels are necessary to its inhibitors. In this context, the design of compounds that target the active and pterin-binding site has been considered an important strategy (Figure 11—
In addition, with the goal to identify the differences among bNOS and other isoforms, crystallography studies were performed using different inhibitor chemotypes. Researchers observed that Tyr706 from nNOS is conserved in bNOS (Tyr 357) and both have the same rotameric behavior, which is very different, compared with eNOS. This molecular feature can be useful to design new selective bNOS over eNOS inhibitor. Since the pharmacokinetic properties are very different between bNOS and nNOS, selectivity over the latter is not a trouble. However, due to steric hindrance in the tail end of thiophenecarboximidamide analogs, this scaffold can bind differently to bNOS comparatively to nNOS .
3. Clinical studies
A nonselective compound l-NMMA (Figure 12), also known as tilarginine, was evaluated clinically in Translational Research Investigating Underlying Disparities in Acute Myocardial Infarction Patients’ Health Status (TRIUMPH) study in North America and Europe with planned enrollment of 658 patients at 130 centers. The period of study was between January 2005 and August 2006 (the study was terminated early). Using 1 mg/kg bolus and 5 h infusion did not decrease the mortality rates in patients with refractory cardiogenic shock complicating myocardial infarction despite an open infarct artery. Although the good results showed in phase II, it has failed in phase III [70, 71]. In another study l-NMMA resulted in no differences in mean arterial pressure (MAP) after 2 h compared with placebo group .
Evaluating another inhibitor, N(G)-nitro-l-arginine methyl ester (Figure 12—l-NAME), in the treatment of refractory cardiogenic shock, the death at 1 month was 27% in the l-NAME group versus 67% in the control group . Additional studies have been performed to further examination, concluding that TRIUMPH strongly indicated that nonselective NOS inhibitors are not clinically interesting .
Recent phase I study in advanced solid tumors with the iNOS inhibitor
Many scaffolds have been found to inhibit nitric oxide synthases. Some of them were presented in this chapter as promising for important therapeutic activity. It must be emphasized that the research about nitric oxide synthase inhibitors has expressively advanced thanks to the X-ray crystallographic studies of this enzyme. This helps the structure-based design approach toward the search for selective inhibitors of this enzyme and the comprehension of their mechanism of action. Notwithstanding, efforts have been made for imparting them with a drug-like profile.
The authors thank CAPES, for RAM Serafim scholarship, and CNPq, for EI Ferreira fellowship.
Kerwin JF, Lancaster JR, Feldman PL. Nitric oxide: a new paradigm for second messengers. Journal of Medicinal Chemistry. 1995; 38:4343–4362. DOI: 10.1021/jm00022a001
Al-sadoni HH, Ferro A. S-nitrosothiols: a class of nitric oxide-donor drugs. Clinical Science. 2000; 98:507–520. DOI: 10.1042/cs0980507
Daff S. NO synthase: structures and mechanisms. Nitric Oxide. 2010; 23:1–11. DOI: 10.1016/j.niox.2010.03.001
Serafim RAM, Primi MC, Trossini GHG, Ferreira EI. Nitric oxide: state of the art in drug design. Current Medicinal Chemistry. 2012; 19:386–405. DOI: 10.2174/092986712803414321
Hobbs AJ, Higgs A, Moncada S. Inhibition of nitric oxide synthase as a potential therapeutic target. Annual Review of Pharmacology and Toxicology. 1999; 39:191–220. DOI: 10.1146/annurev.pharmtox.39.1.191
Rehni AK, Singh TG, Kalra R, Singh N. Pharmacological inhibition of inducible nitric oxide synthase attenuates the development of seizure in mice. Nitric Oxide. 2009; 21:120–125. DOI: 10.1016/j.niox.2009.06.001
Kavya R, Saluja R, Singh S, Dikshit M. Nitric oxide synthase regulation and diversity: implications in Parkinson's disease. Nitric Oxide. 2006; 15:280–294. DOI: 10.1016/j.niox.2006.07.003
Zheng B, Zheng T, Wang L, Chen X, Shi C, Zhao S. Aminoguanidine inhibition of iNOS activity ameliorates cerebral vasospasm after subarachnoid hemorrhage in rabbits via restoration of dysfunctional endothelial cells. Journal of the Neurological Sciences. 2010; 295:97–103. DOI: 10.1016/j.jns.2010.04.012
Ricciardolo FLM, Nijkamp FP, Folkerts G. Nitric oxide synthase (NOS) as therapeutic target for asthma and chronic obstructive pulmonary disease. Current Drug Targets. 2006; 7:721–735. DOI: 10.2174/138945006777435290
Alderton WK, Cooper CE, Knowles RG. Nitric oxide synthases: structure, function and inhibition. Biochemical Journal. 2001; 357:593–615. DOI: 10.1042/bj3570593
Mukherjee P, Cinelli MA, Kang S, Silverman RB. Development of nitric oxide synthase inhibitors for neurodegeneration and neuropathic pain. Chemical Society Reviews. 2014; 43:6814–6838. DOI: 10.1039/c3cs60467e
Vallance P, Leiper J. Blocking NO synthesis: how, where and why? Nature Review Drug Discovery. 2002; 1:939–950. DOI: 10.1038/nrd960
Vallance, P. Nitric oxide: therapeutic opportunities. Fundamental and Clinical Pharmaco-logy. 2003; 17:1–10. DOI: 10.1046/j.1472-8206.2003.00124.x
Yang Z, Misner B, Ji H, Poulos TL, Silverman RB, Meyskens FL, Yang S. Targeting nitric oxide signaling with nNOS inhibitors as a novel strategy for the therapy and prevention of human melanoma. Antioxidants and Redox Signaling. 2013; 19:433–447. DOI: 10.1089/ars.2012.4563
Silverman RB. Design of selective neuronal nitric oxide synthase inhibitors for the prevention and treatment of neurodegenerative diseases. Accounts of Chemical Research. 2009; 42:439–451. DOI: 10.1021/ar800201v
Annedi SC. Cell-permeable inhibitors of neuronal nitric oxide synthase open new prospects for the treatment of neurological disorders. Journal of Medicinal Chemistry. 2015; 58:1064–1066. DOI: 10.1021/acs.jmedchem.5b00057
Paige J, Jaffrey SR. Pharmacologic manipulation of nitric oxide signaling: targeting NOS dimerization and protein-protein interactions. Current Topics in Medicinal Chemistry. 2007; 7:97–114. DOI: 10.2174/156802607779318253
Groves JT, Wang CCY. Nitric oxide synthase: models and mechanism. Current Opinion in Chemical Biology. 2000; 4:687–695. DOI: 10.1016/S1367-5931(00)00146-0
Poulos TL, Li H. Structural basis for isoform-selective inhibition in nitric oxide synthase. Accounts of Chemical Research. 2013; 46:390–398. DOI: 10.1021/ar300175n
Garvey EP, Oplinger JA, Tanoury GJ, Sherman PA, Fowler M, Marshall S, Harmon MF, Paith JE, Furfine ES. Potent and selective inhibition of human nitric oxide synthases. Inhibition by non-amino acid isothioureas. The Journal of Biological Chemistry. 1994; 269:26669–26676.
Garvey EP, Oplinger JA, Furfine ES, Kiff RJ, Laszlo F, Whittle BJ, Knowles RG. 1400W is a slow, tight binding, and highly selective inhibitor of inducible nitric oxide synthase in vitro and in vivo. The Journal of Biological Chemistry. 1997; 272:4959–4963.
Hoffman RA, Nussler NC, Gleixner SL, Zhang G, Ford HR, Langrehr JM, Demetris AJ, Simmons RL. Attenuation of lethal graft-versus-host disease by inhibition of nitric oxide synthase. Transplantation. 1997; 63:94–100.
Anaeigoudari A, Soukhtanloo M, Reisi P, Beheshti F, Hosseini M. Inducible nitric oxide inhibitor aminoguanidine, ameliorates deleterious effects of lipopolysaccharide on memory and long term potentiation in rat. Life Sciences. 2016; 158:22–30. DOI: 10.1016/j.lfs.2016.06.019
Farhad AR, Razavi S, Jahadi S, Saatchi M. Use of aminoguanidine, a selective inducible nitric oxide synthase inhibitor, to evaluate the role of nitric oxide in periapical inflammation. Journal of Oral Science. 2011; 53:225–230.
Soliman MM. Effects of aminoguanidine, a potent nitric oxide synthase inhibitor, on myocardial and organ structure in a rat model of hemorrhagic shock. Journal of Emergencies, Trauma and Shock. 2014;7: 190–195. DOI: 10.4103/0974–2700.136864
Hagmann WK, Caldwell CG, Chen PL, Durette PL, Esser CK, Lanza TJ, Kopka IE, Guthikonda R, Shah SK, Maccoss M, Chabin RM, Fletcher D, Grant SK, Green BG, Humes JL, Kelly TM, Luell S, Meurer R, Moore V, Pacholok SG, Pavia T, Williams HR, Wong KK. Substituted 2-aminopyridines as inhibitors of nitric oxide synthases. Bioorganic and Medicinal Chemistry. 2000; 10:1975–1978. DOI: 10.1016/S0960-894X(00)00389-9
Connolly S, Aberg A, Arvai A, Beaton HG, Cheshire DR, Cook AR, Cooper S, Cox D, Hamley P, Mallinder P, Millichip I, Nicholls DJ, Rosenfeld RJ, St-Gallay SA, Tainer J, Tinker AC, Wallace AV. 2-aminopyridines as highly selective inducible nitric oxide synthase inhibitors. Differential binding modes dependent on nitrogen substitution. Journal of Medicinal Chemistry. 2004; 47:3320–3323. DOI: 10.1021/jm031035n
Tinker AC, Beaton HG, Boughton-Smith N, Cook TR, Cooper SL, Fraser-Rae L, Hallam K, Hamley P, Mcinally T, Nicholls DJ, Pimm AD, Wallace AV. 1,2-Dihydro-4-quinazolinamines: potent, highly selective inhibitors of inducible nitric oxide synthase which show antiinflammatory activity in vivo. Journal of Medicinal Chemistry. 2003; 46:913–916. DOI: 10.1021/jm0255926
Garcin ED, Arvai AS, Rosenfeld RJ, Kroeger MD, Crane BR, Andersson G, Andrews G, Hamley PJ, Mallinder PR, Nicholls DJ, St-Gallay SA, Tinker AC, Gensmantel NP, Mete A, Cheshire DR, Connolly S, Stuehr DJ, Aberg A, Wallace AV, Tainer JA, Getzoff ED. Anchored plasticity opens doors for selective inhibitor design in nitric oxide synthase. Nature Chemical Biology. 2008; 4:700–707. DOI: 10.1038/nchembio.115
Sharma MC, Sharma S. Investigation on quantitative structure activity relationships of a series of inducible nitric oxide. Interdisciplinary Sciences. DOI: 10.1007/s12539-016-0176-5
Maccallini C, Montagnani M, Paciotti R, Ammazzalorso A, De Filippis B, Di Matteo M, Di Silvestre S, Fantacuzzi M, Giampietro L, Potenza MA, Re N, Pandolfi A, Amoroso R. Selective acetamidine-based nitric oxide synthase inhibitors: synthesis, docking, and biological studies. ACS Medicinal Chemistry Letters. 2015; 6:635–640. DOI: 10.1021/acsmedchemlett.5b00149
Yap BK, Harjani JR, Leung EW, Nicholson SE, Scanlon MJ, Chalmers DK, Thompson PE, Baell JB, Norton RS. Redox-stable cyclic peptide inhibitors of the SPSB2-iNOS interactions. FEBS Letters. 2016; 590:696–704. DOI: 10.1002/1873-3468.12115
Harjani JR, Yap BK, Leung EWW, Lucke A, Nicholson SE, Scanlon MJ, Chalmers DK, Thompson PE, Norton RS, Baell JB. Design, synthesis, and characterization of cyclic peptidomimetics of the inducible nitric oxide synthase binding epitope that disrupt the protein–protein interaction involving SPRY domain-containing suppressor of cytokine signaling box protein (SPSB) 2 and inducible nitric oxide synthase. Journal of Medicinal Chemistry. 2016; 59: 5799–5809. DOI: 10.1021/acs.jmedchem.6b00386
Bonnefous C, Payne JE, Roppe J, Zhuang H, Chen X, Symons KT, Nguyen PM, Sablad M, Rozenkrants N, Zhang Y, Wang L, Severance D, Walsh JP, Yazdani N, Shiau AK, Noble SA, Rix P, Rao TS, Hassig CA, Smith ND. Discovery of inducible nitric oxide synthase (iNOS) inhibitor development candidate KD7332, part 1: identification of a novel, potent, and selective series of quinolinone iNOS dimerization inhibitors that are orally active in rodent pain models. Journal of Medicinal Chemistry. 2009; 52:3047–3062. DOI: 10.1021/jm900173b
Payne JE, Bonnefous C, Symons KT, Nguyen PM, Sablad M, Rozenkrants N, Zhang Y, Wang L, Yazdani N, Shiau AK, Noble SA, Rix P, Rao TS, Hassig CA, Smith ND. Discovery of dual inducible/neuronal nitric oxide synthase (iNOS/nNOS) inhibitor development candidate 4-((2-cyclobutyl-1H-imidazo[4,5-b]pyrazin-1yl)methyl)-7,8-difluoroquinolin-2(1H)-one (KD7332) part 2: identification of a novel, potent, and selective series of benzimidazole-quinolinone iNOS/nNOS dimerization inhibitors that are orally active in pain models. Journal of Medicinal Chemistry. 2010; 53:7739–7755. DOI: 10.1021/jm100828n
Cara LCL, Camacho ME, Carrión MD, Tapias V, Gallo MA, Escames G, Acuña-Castroviejo D, Espinosa A, Entrena A. Phenylpyrrole derivatives as neuronal and inducible nitric oxide synthase (nNOS and iNOS) inhibitors. European Journal of Medicinal Chemistry. 2009; 44:2655–2666. DOI: 10.1016/j.ejmech.2008.11.013
Nieto CI, Cabildo MP, Cornago MP, Sanz D, Claramunt RM, Torralba MC, Torres MR, Elguero J, García JA, López A, Acuña-Castroviejo D. Fluorination effects on NOS inhibitory activity of pyrazoles related to curcumin. Molecules. 2015; 20:15643–15665. DOI: 10.3390/molecules200915643
Chayah M, Carrión MD, Gallo MA, Jiménez R, Duarte J, Camacho ME. Development of urea and thiourea kynurenamine derivatives: synthesis, molecular modeling, and biological evaluation as nitric oxide synthase inhibitors. ChemMedChem. 2015; 10:874–882. DOI: 10.1002/cmdc.201500007
Li W, Zheng S, Higgins M, Morra Jr RP, Mendis AT, Chien CW, Ojima I, Mierke DF, Dinkova-Kostova AT, Honda T. New monocyclic, bicyclic, and tricyclic ethynylcyanodienones as activators of the Keap1/Nrf2/ARE pathway and inhibitors of inducible nitric oxide synthase. Journal of Medicinal Chemistry. 2015; 58:4738–4748. DOI: 10.1021/acs.jmedchem.5b00393
Tang W, Li H, Poulos TL, Silverman RB. Mechanistic studies of inactivation of inducible nitric oxide synthase by amidines. Biochemistry. 2015; 54:2530–2538. DOI: 10.1021/acs.biochem.5b00135
Yin GP, Li LC, Zhang QZ, An YW, Zhu JJ, Wang ZM, Chou GX, Wang ZT. iNOS inhibitory activity of sesquiterpenoids and a monoterpenoid from the rhizomes of Curcuma wenyujin. Journal of Natural Products. 2014; 77:2161–2169. DOI: 10.1021/np400984c
Lee J, Kim H, Lee TG, Yang I, Won DH, Choi H, Nam SJ, Kang H. Anmindenols A and B, inducible nitric oxide synthase inhibitors from a marine-derived Streptomycessp. Journal of Natural Products. 2014; 77:1528–1531. DOI: 10.1021/np500285a
Brozíckova C, Mikulecka A, Otáhal J. Effect of 7-nitroindazole, a neuronal nitric oxide synthase inhibitor, on behavior and physiological parameters. Physiological Research. 2014; 63:637–648.
Banach M, Piskorska B, Czuczwar SJ, Borowicz KK, Nitric oxide, epileptic seizures, and action of antiepileptic drugs. CNS Neurological Disorders Drug Discovery. 2011; 10:808–819. DOI: 10.2174/187152711798072347.
Entrena A, Camacho E, Carrión D, López-Cara LC, Velasco G, León J, Escames G, Acuña-Castroviejo D, Tapias V, Gallo MA, Vivo A, Espinosa A. Kynurenamines as neural nitric oxide synthase inhibitors. Journal of Medicinal Chemistry. 2005; 48:8174–8181. DOI: 10.1021/jm050740o
Maccallini C, Patruno A, Lannutti F, Ammazzalorso A, Filippis BD, Fantacuzzi M, Franceschelli S, Giampietro L, Masella S, Felaco M, Re N, Amoroso R. N-substituted acetamidine and 2-methylimidazole derivates as selective inhibitors of neuronal nitric oxide synthase. Bioorganic and Medicinal Chemistry Letters. 2010; 20:6495–6499. DOI: 10.1016/j.bmcl.2010.09.059
Delker SL, Ji H, Li H, Jamal J, Fang J, Xue F, Silverman RB, Poulos TL. Unexpected binding modes of nitric oxide synthase inhibitors effective in the prevention of cerebral palsy phenotype in an animal model. Journal of the American Chemical Society. 2010; 132:5437–5442. DOI: 10.1021/ja910228a
Lawton GR, Ranaivo HR, Wing LK, Ji H, Xue F, Martesek P, Roman LJ, Watterson DM, Silverman RB. Analogues of 2-aminopyridine-based selective inhibitors of neuronal nitric oxide synthase with increased bioavailability. Bioorganic and Medicinal Chemistry. 2009; 17:2371–2380. DOI: 10.1016/j.bmc.2009.02.017
Silverman RB, Lawton GR, Ranaivo HR, Chico LK, Seo J, Watterson DM. Effect of potential amine prodrugs of selective neuronal nitric oxide synthase inhibitors on blood-brain barrier penetration. Bioorganic and Medicinal Chemistry. 2009; 17:7593–7605. DOI: 10.1016/j.bmc.2009.08.065
Xue F, Fang J, Lewis WW, Martásek P, Roman LJ, Silverman RB. Potent and selective neuronal nitric oxide synthase inhibitors with improved cellular permeability. Bioorganic and Medicinal Chemistry Letters. 2010; 20:554–557. DOI: 10.1016/j.bmcl.2009.11.086
Carrión D, Chayah M, Entrena A, López A, Gallo MA, Acuña-Castroviejo D, Camacho ME. Synthesis and biological evaluation of 4,5-dihydro-1H-pyrazole derivatives as potential nNOS/iNOS selective inhibitors. Part 2: influence of diverse substituents in both the phenyl moiety and the acyl group. Bioorganic and Medicinal Chemistry. 2013; 21:4132–4142. DOI: 10.1016/j.bmc.2013.05.016
Cinelli MA, Li H, Chreifi G, Martásek P, Roman LJ, Poulos TL, Silverman RB. Simplified 2‑aminoquinoline-based scaffold for potent and selective neuronal nitric oxide synthase inhibition. Journal of Medicinal Chemistry. 2014; 57:1513–1530. DOI: 10.1021/jm401838x
Cinelli MA, Li H, Pensa AV, Kang S, Roman LJ, Martásek P, Poulos TL, Silverman RB. Phenyl ether- and aniline-containing 2‑aminoquinolines as potent and selective inhibitors of neuronal nitric oxide synthase. Journal of Medicinal Chemistry. 2015; 58:8694–8712. DOI: 10.1021/acs.jmedchem.5b01330
Mukherjee P, Li H, Sevrioukova I, Chreifi G, Martásek P, Roman LJ, Poulos TL, Silverman RB. Novel 2,4-disubstituted pyrimidines as potent, selective, and cell-permeable inhibitors of neuronal nitric oxide synthase. Journal of Medicinal Chemistry. 2015; 58:1067-1088. DOI: 10.1021/jm501719e
Li H, Wang HY, Kang S, Silverman RB, Poulos TL. Electrostatic control of isoform selective inhibitor binding in nitric oxide synthase. Biochemistry. 2016; 55:3702–3707. DOI: 10.1021/acs.biochem.6b00261
Kang S, Li H, Tang W, Martasek P, Roman LJ, Poulos TL, Silverman RB. 2-aminopyridines with a truncated side chain to improve human neuronal nitric oxide synthase inhibitory potency and selectivity. Journal of Medicinal Chemistry. 2015; 58:5548–5560. DOI: 10.1021/acs.jmedchem.5b00573
Wang HY, Qin Y, Li H, Roman LJ, Martasek P, Poulos TL, Silverman RB. Potent and selective human neuronal nitric oxide synthase inhibition by optimization of the 2-aminopyridine-based scaffold with a pyridine linker. Journal of Medicinal Chemistry. 2016; 59:4913–4925. DOI: 10.1021/acs.jmedchem.6b00273
Xu G, Chen Y, Shen K, Wang X, Li F, He Y. The discovery of potentially selective human neuronal nitric oxide synthase (nNOS) inhibitors: a combination of pharmacophore modelling, CoMFA, virtual screening and molecular docking studies. International Journal of Molecular Sciences. 2014; 15:8553–8569. DOI: 10.3390/ijms15058553
Xue F, Fang J, Delker SL, Li H, Martasek P, Roman LJ, Poulos TL, Silverman RB. Symmetric double-headed aminopyridines, a novel strategy for potent and membrane-permeable inhibitors of neuronal nitric oxide synthase. Journal of Medicinal Chemistry. 2011; 54:2039–2048. DOI: 10.1021/jm101071n
Delker SL, Xue F, Li H, Jamal J, Silverman RB, Poulos TL. Role of zinc in isoform-selective inhibitor binding to neuronal nitric oxide synthase. Biochemistry. 2010; 49:10803–10810. DOI: 10.1021/bi1013479
Huang H, Li H, Martásek P, Roman LJ, Poulos TL, Silverman RB. Structure-guided design of selective inhibitors of neuronal nitric oxide synthase. Journal of Medicinal Chemistry. 2013; 56:3024–3032. DOI: 10.1021/jm4000984
Jing Q, Li H, Chreifi G, Roman LJ, Martásek P, Poulos TL, Silverman RB. Chiral linkers to improve selectivity of double-headed neuronal nitric oxide synthase inhibitors. Bioorganic and Medicinal Chemistry Letters. 2013; 23:5674–5679. DOI: 10.1016/j.bmcl.2013.08.034
Jing Q, Li H, Roman LJ, Martásek P, Poulos TL, Silverman RB. Accessible chiral linker to enhance potency and selectivity of neuronal nitric oxide synthase inhibitors. ACS Medicinal Chemistry Letters. 2014; 5:56–60. DOI: 10.1021/ml400381s
Kang S, Tang W, Li H, Chreifi G, Martásek P, Roman LJ, Poulos TL, Silverman RB. Nitric oxide synthase inhibitors that interact with both heme propionate and tetrahydrobiopterin show high isoform selectivity. Journal of Medicinal Chemistry. 2014; 57:4382–4396. DOI: 10.1021/jm5004182
Huang H, Li H, Yang S, Chreifi G, Martásek P, Roman LJ, Meyskens FL, Poulos TL, Silverman RB. Potent and selective double-headed thiophene-2-carboximidamide inhibitors of neuronal nitric oxide synthase for the treatment of melanoma. Journal of Medicinal Chemistry. 2014; 57;686–700. DOI: 10.1021/jm401252e
Gusarov I, Shatalin K, Starodubtseva M, Nudler E. Endogenous nitric oxide protects bacteria against a wide spectrum of antibiotics. Science. 2009; 325:1380–1384. DOI: 10.1126/science.1175439.
Holden JK, Li H, Jing Q, Kang S, Richo J, Silverman RB, Poulos TL. Structural and biological studies on bacterial nitric oxide synthase inhibitors. Proceedings of the National Academy of Sciences of the United States of America. 2013; 110:18127–12131. DOI: 10.1073/pnas.1314080110
Holden JK, Kang S, Hollingsworth SA, Li H, Lim N, Chen S, Huang H, Xue F, Tang W, Silverman RB, Poulos TL. Structure-based design of bacterial nitric oxide synthase inhibitors. Journal of Medicinal Chemistry. 2015; 58: 994–1004. DOI: 10.1021/jm501723p
Holden JK, Dejam D, Lewis MC, Huang H, Kang S, Jing Q, Xue F, Silverman RB, Poulos TL. Inhibitor bound crystal structures of bacterial nitric oxide synthase. Biochemistry. 2015; 54:4075–4082. DOI: 10.1021/acs.biochem.5b00431.
Alexander JH, Reynolds HR, Stebbins AL, Dzavik V, Harrington RA, Van der Werf F, Hechman JS.Effect of tilarginine acetate in patients with acute myocardial infarction and cardiogenic shock: the TRIUMPH randomized controlled trial. Journal of American Medical Association. 2007; 297:1657–1666.
Wong VW, Lerner E. Nitric oxide inhibition strategies. Future Sciences OA. 2015; 1:pii-FSO35. DOI: 10.4155/FSO.15.35
Dzavik V, Cotter G, Reynolds HR, Alexander JH, Ramanathan K, Stebbis AL, Hathaway D, Farkouh ME, Ohman EM, Baran DA, Prondzinsky R, Panza JA, Cantor WJ, Vered Z, Buller CE, Kleiman NS, Webb JG, Holmes DR, Parrillo JE, Hazen SL, Gross SS, Harrington RA, Hachman JS. Effect of nitric oxide synthase inhibition on haemodynamics and outcome of patients with persistent cardiogenic shock complicating acute myocardial infarction: a phase II dose-ranging study. European Heart Journal. 2007; 28:1109–1116. DOI: 10.1093/eurheartj/ehm075.
Cottera G, Kaluskia E, Miloa O, Blatta A, Salaha A, Hendlera A, Krakovera R, Golickb A, Vereda Z. LINCS: L-NAME (a NO synthase inhibitor) in the treatment of refractory cardiogenic shock: a prospective randomized study. European Heart Journal. 2003; 24:1287–1295. DOI: 10.1016/S0195-668X(03)00193-3.
Salem R, Mebazaa A. Nitric oxide inhibition rapidly increases blood pressure with no change in outcome in cardiogenic shock: the TRIUMPH trial. Critical Care. 2007; 11;1–2. DOI: 10.1186/cc5925.
Luke JJ, LoRusso P, Shapiro GI, Krivoshik A, Schuster R, Yamazaki T, Arai Y, Fakhoury A, Dmuchowski C, Infante JR. ASP9853, an inhibitor of inducible nitric oxide synthase dimerization, in combination with docetaxel: preclinical investigation and a Phase I study in advanced solid tumors. Cancer Chemotherapy Pharmacology. 2016; 77:549–558. DOI: 10.1007/s00280-016-2967-0.