Tethered Dimers as NAD Synthetase Inhibitors with Antibacterial Activity

Velu, Sadanandan E.; Cristofoli, Walter A.; Garcı́a, Gabriel; Brouillette, Christie G.; Pierson, Milton C.; Luan, Chi Hao; DeLucas, Lawrence J.; Brouillette, Wayne J.

doi:10.1021/jm030003x

Cited by 24 publications

(42 citation statements)

References 21 publications

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“…The indole derivatives are substrate analogs with two aromatic groups linked by an aliphatic chain meant to mimic the pyridine nucleotide and adenine of nicotinic acid adenine dinucleotide, and have been reported previously to have IC 50 values of 9 M to Ͼ100 M against the Bacillus subtilis NadE enzyme (31). These compounds had also demonstrated MIC values ranging from 1.5 M to Ͼ50 M for several Gram-positive organisms including B. subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus enteritidis.…”

Section: Discussionmentioning

confidence: 99%

“…Synthesis of Inhibitors and Inhibition Assays-NAD synthetase inhibitors (Table 1) were synthesized as described previously, and analytical data consistent with the published data were obtained for the final purified products (31). For IC 50 determination, the NAD synthetase reaction was conducted as described (32) with a total volume of 50 l in a 96-well white flat bottom 1 ⁄ 2-area plate.…”

Section: Methodsmentioning

confidence: 99%

“…Interruption of NAD Synthesis Is Bactericidal for Growing Cells of M. tuberculosis-To explore further the consequences of sudden disruption of NAD levels, we synthesized several compounds that had been reported previously to be effective inhibitors of NAD synthetase (NadE) in several Gram-positive organisms (31). Two of these inhibitors showed modest aerobic growth inhibition at 19.5 and 17.9 M ( Table 1).…”

Section: Tuberculosis Pncb1 and Pncb2mentioning

confidence: 99%

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Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

Boshoff

Tahlan

et al. 2008

Journal of Biological Chemistry

146

168

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Despite the presence of genes that apparently encode NAD salvage-specific enzymes in its genome, it has been previously thought that Mycobacterium tuberculosis can only synthesize NAD de novo. Transcriptional analysis of the de novo synthesis and putative salvage pathway genes revealed an up-regulation of the salvage pathway genes in vivo and in vitro under conditions of hypoxia. [14 C]Nicotinamide incorporation assays in M. tuberculosis isolated directly from the lungs of infected mice or from infected macrophages revealed that incorporation of exogenous nicotinamide was very efficient in in vivo-adapted cells, in contrast to cells grown aerobically in vitro. Two putative nicotinic acid phosphoribosyltransferases, PncB1 (Rv1330c) and PncB2 (Rv0573c), were examined by a combination of in vitro enzymatic activity assays and allelic exchange studies. These studies revealed that both play a role in cofactor salvage. Mutants in the de novo pathway died upon removal of exogenous nicotinamide during active replication in vitro. Cell death is induced by both cofactor starvation and disruption of cellular redox homeostasis as electron transport is impaired by limiting NAD. Inhibitors of NAD synthetase, an essential enzyme common to both recycling and de novo synthesis pathways, displayed the same bactericidal effect as sudden NAD starvation of the de novo pathway mutant in both actively growing and nonreplicating M. tuberculosis. These studies demonstrate the plasticity of the organism in maintaining NAD levels and establish that the two enzymes of the universal pathway are attractive chemotherapeutic targets for active as well as latent tuberculosis.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Tuberculosis Pncb1 and Pncb2mentioning

confidence: 99%

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Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

Boshoff

Tahlan

et al. 2008

Journal of Biological Chemistry

146

168

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“…Finally, NAD ϩ synthetase activity is not required to recycle nicotinamide in humans, and its central role in recycling in microbes suggests that it may be an excellent target for antimicrobials. NAD ϩ synthetase inhibitors have proven antibiotic properties, killing Gram-positive bacteria (Velu et al, 2003).…”

Section: Prospects For Drugs Targeted To Inhibition Of Nad ؉ Biosynthmentioning

confidence: 99%

NAD⁺and Vitamin B₃: From Metabolism to Therapies

Sauve

2007

J Pharmacol Exp Ther

283

205

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The role of NAD ϩ metabolism in health and disease is of increased interest as the use of niacin (nicotinic acid) has emerged as a major therapy for treatment of hyperlipidemias and with the recognition that nicotinamide can protect tissues and NAD ϩ metabolism in a variety of disease states, including ischemia/reperfusion. In addition, a growing body of evidence supports the view that NAD ϩ metabolism regulates important biological effects, including lifespan. NAD ϩ exerts potent effects through the poly(ADP-ribose) polymerases, mono-ADPribosyltransferases, and the recently characterized sirtuin enzymes. These enzymes catalyze protein modifications, such as ADP-ribosylation and deacetylation, leading to changes in protein function. These enzymes regulate apoptosis, DNA repair, stress resistance, metabolism, and endocrine signaling, suggesting that these enzymes and/or NAD ϩ metabolism could be targeted for therapeutic benefit. This review considers current knowledge of NAD ϩ metabolism in humans and microbes, including new insights into mechanisms that regulate NAD ϩ biosynthetic pathways, current use of nicotinamide and nicotinic acid as pharmacological agents, and opportunities for drug design that are directed at modulation of NAD ϩ biosynthesis for treatment of human disorders and infections.Vitamin B 3 (nicotinamide and nicotinic acid) is essential to all living cells. Vitamin B 3 is biosynthetically converted to nicotinamide adenine dinucleotide (Fig. 1, NAD ϩ ), a versatile acceptor of hydride equivalents to form the reduced dinucleotide, NADH. The phosphorylated forms of the nicotinamide dinucleotides (NADP/NADPH) perform similar chemical functions within cells, although these are generally used in biosynthetic pathways and in cell protection mechanisms against reactive oxygen species. NAD(P)H provides reducing equivalents for cellular biochemistry and energy metabolism. Within eukaryotic cells, energy metabolism is largely mediated by electron transport chains found within the mitochondrion, and NADH plays a vital role in furnishing reducing equivalents to fuel oxidative phosphorylation. Thus, cellular energy metabolism is substantially mediated by vitamin B 3 -derived cofactors, and a large fraction of anabolic and catabolic pathways incorporates these cofactors within them.Nicotinamide dinucleotides also react as electrophiles to transfer the ADP-ribose (ADPR) moiety to a nucleophile. ADPR transfer to small nucleophiles forms ADPR (nucleophile/water), cyclic-ADPR (nucleophile/N1 adenine), and nicotinic acid adenine dinucleotide-phosphate (derived from NADP ϩ , nucleophile/nicotinic acid). These compounds have been shown to regulate processes, such as channel opening and calcium release (Pollak et al., 2007). Furthermore, ADPR transfer modifies protein nucleophilic side chains (Hassa et al., 2006). The ADPR-transfer enzymes fall into three distinct families, the mono-ADP-ribosyltransferases (Hassa et al., 2006), the poly(ADP-ribosyl) polymerases (PARPs) (Virag and Szabo, 2002;Hassa et al., 200...

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“…Inhibitors of B. subtilis NAD showing promising antibacterial activity have been published [37,38] and the M. tuberculosis enzyme was shown to be essential for the growth of M. Tuberculosis [12]. We consider NAD synthetase as an exploitable target for the identification of novel antitubercular agents [39][40][41].…”

Section: Introductionmentioning

confidence: 99%

Signalling Inhibitors Against Mycobacterium tuberculosis – Early Days of a New Therapeutic Concept in Tuberculosis

Hegymegi-Barakonyi¹,

Székely

Varga³

et al. 2008

CMC

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Tuberculosis causes nearly two million deaths per year world-wide. In addition multidrug-resistant mycobacterial strains rapidly emerge so novel therapeutic approaches are needed. Recently, several promising mycobacterial target molecules were identified, which are involved in bacterial or host cell signalling e.g. the serine/threonine protein kinases, PknB and PknG, NAD kinase and the NAD synthetase. Here we describe some early efforts in the development of novel signal transduction inhibitory anti-mycobacterial drugs using a multiple target approach, with special emphasis on the kinase inhibitory field. Initially, we are using the Nested Chemical Library (NCL) technology and pharmacophore modelling. A hit-finding library, consisting of approximately 19000 small molecules with a bias for prototypic kinase inhibitors from our NCL library and commercial sources was virtually screened against these validated target molecules. Protein structures for the virtual screening were taken from the published three dimensional crystal structures of the enzymes. The hits from the virtual screening were subsequently tested in enzymatic assay systems. Potent hits were then tested for biological activity in macrophages, infected with mycobacteria. The final goal of this exercise is not only to identify potent anti-mycobacterial substances, but also a common pharmacophore for the mycobacterial target PknG in combination with PknB, NAD kinase and/or NAD synthetase. This common pharmacophore still needs to be a unique pharmacophore for the mycobacterial target proteins over human off-targets. Such a pharmacophore might then drive the optimization of a completely new profile of an antibiotic agent with activity against latent mycobacteria and resistance mycobacterial strains.

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Tethered Dimers as NAD Synthetase Inhibitors with Antibacterial Activity

Cited by 24 publications

References 21 publications

Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

NAD⁺and Vitamin B₃: From Metabolism to Therapies

Signalling Inhibitors Against Mycobacterium tuberculosis – Early Days of a New Therapeutic Concept in Tuberculosis

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Tethered Dimers as NAD Synthetase Inhibitors with Antibacterial Activity

Cited by 24 publications

References 21 publications

Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

Biosynthesis and Recycling of Nicotinamide Cofactors in Mycobacterium tuberculosis

NAD+and Vitamin B3: From Metabolism to Therapies

Signalling Inhibitors Against Mycobacterium tuberculosis – Early Days of a New Therapeutic Concept in Tuberculosis

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Product

Resources

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NAD⁺and Vitamin B₃: From Metabolism to Therapies