Involvement of p53 in the cytotoxic activity of the NAMPT inhibitor FK866 in myeloid leukemic cells

Thakur, Basant Kumar; Dittrich, Tino; Prakash, Chandra; Becker, Annette; Kuehnau, Wolfgang; Klusmann, Jan‐Henning; Reinhardt, Dirk; Welte, Karl

doi:10.1002/ijc.27726

Cited by 45 publications

(47 citation statements)

References 60 publications

(120 reference statements)

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“…Break down of ATP levels was associated with increased cell death. In contrast to another study [6], we demonstrated that FK866 reduced NAMPT activity, depleted NAD and ATP content and induced cell death in p53-deficient Hep3B cells suggesting that FK866-mediated cell death does not depend on functional p53. Our results are in line with a study performed in chronic lymphocytic leukemia cells [14].…”

Section: Discussioncontrasting

confidence: 82%

FK866-induced NAMPT inhibition activates AMPK and downregulates mTOR signaling in hepatocarcinoma cells

Schuster

Penke

Gorski

et al. 2015

Biochemical and Biophysical Research Communications

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Section: Discussioncontrasting

confidence: 82%

FK866-induced NAMPT inhibition activates AMPK and downregulates mTOR signaling in hepatocarcinoma cells

Schuster

Penke

Gorski

et al. 2015

Biochemical and Biophysical Research Communications

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“…While Nahimana et al reported an EC 50 value of 7.2 nM using the MTT assay, 18) Thakur et al found no inhibition at 100 nM using the trypan blue exclusion assay. 19) Here, we showed that FK866 inhibits K562 cell growth with high potency using the WST-8 assay, which is similar to the MTT assay. This discrepancy in findings may be due to differences in the mode of measurement.…”

Section: Discussionsupporting

confidence: 61%

“…We confirmed that FK866 did indeed inhibit K562 cell growth using a DNA content-based assay (data not shown). 19) We demonstrated that our hit compounds reduced NAD content in the cytoplasm to about 60% of baseline levels at 5 h after addition of hit compounds and FK866. Previous studies showed time-dependent decreases in NAD induced by FK866 in THP-1 and other cells, 12,17) and our data showed that caspase 3/7 activation in THP-1 occurred after 24 h (data not shown), suggesting that apoptosis was caused by NAD depletion.…”

Section: Discussionmentioning

confidence: 70%

Discovery of a Novel Nicotinamide Phosphoribosyl Transferase (NAMPT) Inhibitor <i>via</i> <i>in Silico</i> Screening

Takeuchi

Niimi

Masumoto

et al. 2014

Biological & Pharmaceutical Bulletin

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Nicotinamide phosphoribosyl transferase (NAMPT) is a key enzyme in the salvage pathway of mammalian nicotinamide adenine dinucleotide (NAD) biosynthesis, catalyzing the synthesis of nicotinamide mononucleotide from nicotinamide (Nam). The diverse functions of NAD suggest that NAMPT inhibitors are potential drug candidates as anticancer agents, immunomodulators, or other agents. However, difficulty in conducting high-throughput NAMPT assay with good sensitivity has hampered the discovery of novel anti-NAMPT drugs with improved profiles. We combined an in silico screening strategy with a radioisotope (RI)-based enzyme assay and rationally identified promising NAMPT inhibitors with novel structures. AS1604498 was the most potent inhibitor, with an IC 50 of 44 nM, and inhibited THP-1 and K562 cell line growth with the IC 50 of 198 nM and 673 nM, respectively. The mode of action was found to reduce intracellular NAD following apoptosis, suggesting that these compounds inhibit NAMPT in cell-based assay. This strategy can be used to discover new drug candidates with targets which are difficult to assess through high-throughput screening. Our hit compounds may be used as seed compounds for developing new therapeutics with NAMPT.Key words nicotinamide phosphoribosyl transferase; in silico screening; FK866; nicotinamide adenine dinucleotide Nicotinamide phosphoribosyl transferase (NAMPT) catalyzes a rate-limiting step in the salvage pathway of mammalian nicotinamide adenine dinucleotide (NAD) biosynthesis. Recently, intracellular NAD has received substantial attention due to the recent discovery that several enzymes, including poly(ADP-ribose) polymerase (PARP) and the Sirtuin-family proteins, use NAD as a substrate, [1][2][3] suggesting that intracellular NAD level may regulate cytokine production, 4) circadian rhythm, 5,6) metabolism, and aging 3,7) through these enzymes. FK866, an NAMPT inhibitor, has been shown to induce apoptosis of cancer cells 8) and immune cells, 9) suggesting the possibility of NAMPT inhibitors as a novel drug class. As such, FK866 and CHS-828, another NAMPT inhibitor, are currently being investigated in clinical trials for cancer treatment.10,11) However, these compounds were discovered by chance using cell-based assay, 12,13) and discovery of new NAMPT inhibitors with better structures has been hindered by the difficulty of conducting high-throughput screening (HTS) with good sensitivity. HTS presents difficulties because a radioisotope (RI)-labeled substrate is needed to retain good assay sensitivity, but a complicated procedure including separation of product from substrate prevents evaluation of large numbers of compounds.Here we report our finding of novel compounds with potent NAMPT inhibitory activity as determined using in silico screening and a 96-well-based sensitive RI-based assay.

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“…156 Cancer cells are more susceptible to NAMPT inhibition than normal cells. 157,158 Clinical studies have demonstrated that serum or blood levels of eNAMPT are increased in patients with cancer and a positive correlation exists between either tissue or eNAMPT levels and the stage of cancer progression. [159][160][161][162][163] However, the molecular pathways of eNAMPT signalling in carcinogenesis are far from clear.…”

Section: Nampt In Cancermentioning

confidence: 98%

Physiological and pathophysiological roles of NAMPT and NAD metabolism

et al. 2015

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Nicotinamide phosphoribosyltransferase (NAMPT) is a regulator of the intracellular nicotinamide adenine dinucleotide (NAD) pool. NAD is an essential coenzyme involved in cellular redox reactions and is a substrate for NAD-dependent enzymes. In various metabolic disorders and during ageing, levels of NAD are decreased. Through its NAD-biosynthetic activity, NAMPT influences the activity of NAD-dependent enzymes, thereby regulating cellular metabolism. In addition to its enzymatic function, extracellular NAMPT (eNAMPT) has cytokine-like activity. Abnormal levels of eNAMPT are associated with various metabolic disorders. NAMPT is able to modulate processes involved in the pathogenesis of obesity and related disorders such as nonalcoholic fatty liver disease (NAFLD) and type 2 diabetes mellitus (T2DM) by influencing the oxidative stress response, apoptosis, lipid and glucose metabolism, inflammation and insulin resistance. NAMPT also has a crucial role in cancer cell metabolism, is often overexpressed in tumour tissues and is an experimental target for antitumour therapies. In this Review, we discuss current understanding of the functions of NAMPT and highlight progress made in identifying the physiological role of NAMPT and its relevance in various human diseases and conditions, such as obesity, NAFLD, T2DM, cancer and ageing.

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Involvement of p53 in the cytotoxic activity of the NAMPT inhibitor FK866 in myeloid leukemic cells

Cited by 45 publications

References 60 publications

FK866-induced NAMPT inhibition activates AMPK and downregulates mTOR signaling in hepatocarcinoma cells

FK866-induced NAMPT inhibition activates AMPK and downregulates mTOR signaling in hepatocarcinoma cells

Discovery of a Novel Nicotinamide Phosphoribosyl Transferase (NAMPT) Inhibitor <i>via</i> <i>in Silico</i> Screening

Physiological and pathophysiological roles of NAMPT and NAD metabolism

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