New assays and approaches for discovery and design of Sirtuin modulators

Expert Opinion on Drug Discovery

Volumn 9, Issue 2, 2014, Pages 183-199

  Schutkowski, Mike ^a   Fischer, Frank ^b   Roessler, Claudia ^a   Steegborn, Clemens ^b  

^a MARTIN LUTHER UNIVERSITY HALLE WITTENBERG   (Germany)

^b UNIVERSITY OF BAYREUTH   (Germany)

Author keywords

Activator; Activity assay; Deacetylases; Deacylases; Drug development; Inhibitor; Mechanism; Modulators; Sirtuin; Substrate specific modulation

Indexed keywords

2,3 NAPHTHALENEDICARBOXALDEHYDE; 3 (3,5 DIBROMO 4 HYDROXYBENZYLIDENE) 1,3 DIHYDRO 5 IODO 2 INDOLONE; 7 AMINO 4 METHYLCOUMARIN; ADENOSINE DIPHOSPHATE RIBOSE; COUMARIN DERIVATIVE; DITHIOTHREITOL; EX 527; FATTY ACID; FLUORESCENT DYE; GLUTAMIC ACID; HISTONE; HISTONE DEACETYLASE INHIBITOR; HYDROLASE INHIBITOR; ISONICOTINAMIDE; NICOTINAMIDASE; NICOTINAMIDE; NICOTINAMIDE ADENINE DINUCLEOTIDE; NICOTINIC ACID; NUCLEOPHILE; POLYPHENOL DERIVATIVE; REDUCED NICOTINAMIDE ADENINE DINUCLEOTIDE; RESVERATROL; SALEMIDE; SIRTINOL; SIRTUIN; SIRTUIN 5; SIRTUIN MODULATING AGENT; SRT 1720; SURAMIN; UNCLASSIFIED DRUG;

ADENOSINE DIPHOSPHATE RIBOSYLATION; ANALYTIC METHOD; APOPTOSIS; BIOASSAY; CAPILLARY ELECTROPHORESIS; CATALYSIS; CHEMICAL ANALYSIS; COMPLEX FORMATION; CRYSTAL STRUCTURE; DEACYLATION; DRUG DESIGN; DRUG DETERMINATION; DRUG POTENCY; DRUG TARGETING; ENZYME ACTIVITY; ENZYME LINKED IMMUNOSORBENT ASSAY; ENZYME SUBSTRATE; FLUORESCENCE SPECTROSCOPY; HIGH PERFORMANCE LIQUID CHROMATOGRAPHY; HISTONE ACETYLATION; HUMAN; IC 50; IN VIVO STUDY; MASS SPECTROMETRY; MICROCHIP ELECTROPHORESIS; MICROFLUIDIC ANALYSIS; NONHUMAN; POLYACRYLAMIDE GEL ELECTROPHORESIS; PRIORITY JOURNAL; PROTEIN ANALYSIS; PROTEIN CONFORMATION; PROTEIN DEACYLATION; PROTEIN FUNCTION; PROTEOMICS; RADIOACTIVITY; REVIEW; STRUCTURE ACTIVITY RELATION; TANDEM MASS SPECTROMETRY; WESTERN BLOTTING;

ANIMALS; BIOLOGICAL ASSAY; DRUG DISCOVERY; HUMANS; SIRTUINS;

EID: 84893149368     PISSN: 17460441     EISSN: 1746045X     Source Type: Journal    
DOI: 10.1517/17460441.2014.875526     Document Type: Review

References (118)

1. Norvell A, McMahon SB. Cell biology. Rise of the rival. Science 2010;327:964-5
2. Rauh D, Fischer F, Gertz M, et al. An acetylome peptide microarray reveals specificities and deacetylation substrates for all human sirtuin isoforms. Nat Commun 2013;4:2327
3. Xu WS, Parmigiani RB, Marks PA. Histone deacetylase inhibitors: molecular mechanisms of action. Oncogene 2007;26:5541-52
4. Cen Y. Sirtuins inhibitors: the approach to affinity and selectivity. Biochim Biophys Acta 2010;1804:1635-44
5. Bell EL, Guarente L. The SirT3 divining rod points to oxidative stress. Mol Cell 2011;42:561-8
6. Guarente L, Picard F. Calorie restrictionthe SIR2 connection. Cell 2005;120:473-82
7. Haigis MC, Sinclair DA. Mammalian sirtuins: biological insights and disease relevance. Annu Rev Pathol 2010;5:253-95
8. Sanchez-Fidalgo S, Villegas I, Sanchez-Hidalgo M, de la Lastra CA. Sirtuin modulators: mechanisms and potential clinical implications. Curr Med Chem 2012;19:2414-41
9. Michan S, Sinclair D. Sirtuins in mammals: insights into their biological function. Biochem J 2007;404:1-13
10. Gertz M, Steegborn C. Function and regulation of the mitochondrial Sirtuin isoform Sirt5 in Mammalia. Biochim Biophys Acta 2010;1804:1658-65
11. Laurent G, German NJ, Saha AK, et al. SIRT4 coordinates the balance between lipid synthesis and catabolism by repressing malonyl CoA decarboxylase. Mol Cell 2013;50:686-98
12. North BJ, Marshall BL, Borra MT, et al. The human Sir2 ortholog, SIRT2, is an NAD+-dependent tubulin deacetylase. Mol Cell 2003;11:437-44
13. Donmez G, Outeiro TF. SIRT1 and SIRT2: emerging targets in neurodegeneration. EMBO Mol Med 2013;5:344-52
14. Blum CA, Ellis JL, Loh C, et al. SIRT1 modulation as a novel approach to the treatment of diseases of aging. J Med Chem 2011;54:417-32
15. Chen L. Medicinal chemistry of sirtuin inhibitors. Curr Med Chem 2011;18:1936-46
16. Sanders BD, Jackson B, Marmorstein R. Structural basis for sirtuin function: what we know and what we don't. Biochim Biophys Acta 2010;1804:1604-16
17. Jin L, Wei W, Jiang Y, et al. Crystal structures of human SIRT3 displaying substrate-induced conformational changes. J Biol Chem 2009;284:24394-405
18. Moniot S, Weyand M, Steegborn C. Structures, substrates, and regulators of Mammalian sirtuins - opportunities and challenges for drug development. Front Pharmacol 2012;3:16
19. Cen Y, Youn DY, Sauve AA. Advances in characterization of human sirtuin isoforms: chemistries, targets and therapeutic applications. Curr Med Chem 2011;18:1919-35
20. Lin H, Su X, He B. Protein lysine acylation and cysteine succination by intermediates of energy metabolism. ACS Chem Biol 2012;7:947-60
21. Du J, Zhou Y, Su X, et al. Sirt5 is a NAD-dependent protein lysine demalonylase and desuccinylase. Science 2011;334:806-9
22. Jiang H, Khan S, Wang Y, et al. SIRT6 regulates TNF-alpha secretion through hydrolysis of long-chain fatty acyl lysine. Nature 2013;496:110-13
23. Liszt G, Ford E, Kurtev M, Guarente L. Mouse Sir2 homolog SIRT6 is a nuclear ADP-ribosyltransferase. J Biol Chem 2005;280:21313-20
24. Du J, Jiang H, Lin H. Investigating the ADP-ribosyltransferase activity of sirtuins with NAD analogues and 32P-NAD. Biochemistry 2009;48:2878-90
25. Grozinger CM, Chao ED, Blackwell HE, et al. Identification of a class of small molecule inhibitors of the sirtuin family of NAD-dependent deacetylases by phenotypic screening. J Biol Chem 2001;276:38837-43
26. Lara E, Mai A, Calvanese V, et al. Salermide, a Sirtuin inhibitor with a strong cancer-specific proapoptotic effect. Oncogene 2009;28:781-91
27. Outeiro TF, Kontopoulos E, Altmann SM, et al. Sirtuin 2 inhibitors rescue alpha-synuclein-mediated toxicity in models of Parkinson's disease. Science 2007;317:516-19
28. Suenkel B, Fischer F, Steegborn C. Inhibition of the human deacylase Sirtuin 5 by the indole GW5074. Bioorg Med Chem Lett 2013;23:143-6
29. Schuetz A, Min J, Antoshenko T, et al. Structural basis of inhibition of the human NAD+-dependent deacetylase SIRT5 by suramin. Structure 2007;15:377-89
30. Trapp J, Meier R, Hongwiset D, et al. Structure-activity studies on suramin analogues as inhibitors of NAD+- dependent histone deacetylases (sirtuins). ChemMedChem 2007;2:1419-31
31. Napper AD, Hixon J, McDonagh T, et al. Discovery of indoles as potent and selective inhibitors of the deacetylase SIRT1. J Med Chem 2005;48:8045-54
32. Gertz M, Fischer F, Nguyen GT, et al. Ex-527 inhibits Sirtuins by exploiting their unique NAD+-dependent deacetylation mechanism. Proc Natl Acad Sci USA 2013;110:E2772-81
33. Howitz KT, Bitterman KJ, Cohen HY, et al. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature 2003;425:191-6
34. Hubbard BP, Gomes AP, Dai H, et al. Evidence for a common mechanism of SIRT1 regulation by allosteric activators. Science 2013;339:1216-19
35. Lakshminarasimhan M, Steegborn C. Emerging mitochondrial signaling mechanisms in physiology, aging processes, and as drug targets. Exp Gerontol 2011;46:174-7
36. Gertz M, Nguyen GT, Fischer F, et al. A molecular mechanism for direct sirtuin activation by resveratrol. PLoS One 2012;7:e49761
37. Sauve AA, Moir RD, Schramm VL, Willis IM. Chemical activation of Sir2- dependent silencing by relief of nicotinamide inhibition. Mol Cell 2005;17:595-601
38. Milne JC, Lambert PD, Schenk S, et al. Small molecule activators of SIRT1 as therapeutics for the treatment of type 2 diabetes. Nature 2007;450:712-16
39. Lakshminarasimhan M, Rauth D, Schutkowski M, Steegborn C. Sirt1 activation by resveratrol is substrate sequence-selective. Aging (Albany NY) 2013;5:151-4
40. Milne JC, Denu JM. The Sirtuin family: therapeutic targets to treat diseases of aging. Curr Opin Chem Biol 2008;12:11-17
41. Gershey EL, Vidali G, Allfrey VG. Chemical studies of histone acetylation. The occurrence of epsilon-N-acetyllysine in the f2a1 histone. J Biol Chem 1968;243:5018-22
42. Sterner R, Vidali G, Allfrey VG. Studies of acetylation and deacetylation in high mobility group proteins. Identification of the sites of acetylation in HMG-1. J Biol Chem 1979;254:11577-83
43. Kim SC, Sprung R, Chen Y, et al. Substrate and functional diversity of lysine acetylation revealed by a proteomics survey. Mol Cell 2006;23:607-18
44. Choudhary C, Kumar C, Gnad F, et al. Lysine acetylation targets protein complexes and co-regulates major cellular functions. Science 2009;325:834-40
45. Zhang J, Sprung R, Pei J, et al. Lysine acetylation is a highly abundant and evolutionarily conserved modification in Escherichia coli. Mol Cell Proteomics 2009;8:215-25
46. Henriksen P, Wagner SA, Weinert BT, et al. Proteome-wide analysis of lysine acetylation suggests its broad regulatory scope in saccharomyces cerevisiae. Mol Cell Proteomics 2012;11:1510-22
47. Hebert A, Dittenhafer-Reed K, Yu W, et al. Calorie restriction and SIRT3 trigger global reprogramming of the mitochondrial protein acetylome. Mol Cell 2013;49:186-99
48. Rardin MJ, Newman JC, Held JM, et al. Label-free quantitative proteomics of the lysine acetylome in mitochondria identifies substrates of SIRT3 in metabolic pathways. Proc Natl Acad Sci USA 2013;110:6601-6
49. Chen Y, Zhao W, Yang JS, et al. Quantitative acetylome analysis reveals the roles of SIRT1 in regulating diverse substrates and cellular pathways. Mol Cell Proteomics 2012;11:1048-62
50. Fritz KS, Galligan JJ, Hirschey MD, et al. Mitochondrial acetylome analysis in a mouse model of alcohol-induced liver injury utilizing SIRT3 knockout mice. J Proteome Res 2012;11:1633-43
51. Morselli E, Marino G, Bennetzen MV, et al. Spermidine and resveratrol induce autophagy by distinct pathways converging on the acetylproteome. J Cell Biol 2011;192:615-29
52. Borra MT, Denu JM. Quantitative assays for characterization of the Sir2 family of NAD(+)-dependent deacetylases. Methods Enzymol 2004;376:171-87
53. Fan Y, Scriba GK. Electrophoretically mediated microanalysis assay for sirtuin enzymes. Electrophoresis 2010;31:3874-80
54. Ohla S, Beyreiss R, Scriba GK, et al. An integrated on-chip sirtuin assay. Electrophoresis 2010;31:3263-7
55. Blackwell L, Norris J, Suto CM, Janzen WP. The use of diversity profiling to characterize chemical modulators of the histone deacetylases. Life Sci 2008;82:1050-8
56. Liu Y, Gerber R, Wu J, et al. High-throughput assays for sirtuin enzymes: a microfluidic mobility shift assay and a bioluminescence assay. Anal Biochem 2008;378:53-9
57. Khan AN, Lewis PN. Unstructured conformations are a substrate requirement for the Sir2 family of NADdependent protein deacetylases. J Biol Chem 2005;280:36073-8
58. Marcotte PA, Richardson PL, Guo J, et al. Fluorescence assay of SIRT protein deacetylases using an acetylated peptide substrate and a secondary trypsin reaction. Anal Biochem 2004;332:90-9
59. Tanner KG, Landry J, Sternglanz R, Denu JM. Silent information regulator 2 family of NAD- dependent histone/ protein deacetylases generates a unique product, 1-O-acetyl-ADP-ribose. Proc Natl Acad Sci USA 2000;97:14178-82
60. Jackson MD, Denu JM. Structural identification of 2'- and 3'-O-acetyl- ADP-ribose as novel metabolites derived from the Sir2 family of beta-NAD+- dependent histone/protein deacetylases. J Biol Chem 2002;277:18535-44
61. Khan AN, Lewis PN. Use of substrate analogs and mutagenesis to study substrate binding and catalysis in the Sir2 family of NAD-dependent protein deacetylases. J Biol Chem 2006;281:11702-11
62. McDonagh T, Hixon J, DiStefano PS, et al. Microplate filtration assay for nicotinamide release from NAD using a boronic acid resin. Methods 2005;36:346-50
63. Hoffmann K, Heltweg B, Jung M. Improvement and validation of the fluorescence-based histone deacetylase assay using an internal standard. Arch Pharm (Weinheim) 2001;334:248-52
64. Bedalov A, Gatbonton T, Irvine WP, et al. Identification of a small molecule inhibitor of Sir2p. Proc Natl Acad Sci USA 2001;98:15113-18
65. Luo J, Nikolaev AY, Imai S, et al. Negative control of p53 by Sir2alpha promotes cell survival under stress. Cell 2001;107:137-48
66. Feldman JL, Baeza J, Denu JM. Activation of the protein deacetylase SIRT6 by long-chain fatty acids and widespread deacylation by mammalian sirtuins. J Biol Chem 2013;288(43):31350-6
67. Borra MT, Langer MR, Slama JT, Denu JM. Substrate specificity and kinetic mechanism of the Sir2 family of NAD+-dependent histone/protein deacetylases. Biochemistry 2004;43:9877-87
68. Hoffmann K, Brosch G, Loidl P, Jung M. A non-isotopic assay for histone deacetylase activity. Nucleic Acids Res 1999;27:2057-8
69. Heltweg B, Jung M. A homogeneous nonisotopic histone deacetylase activity assay. J Biomol Screen 2003;8:89-95
70. Feng Y, Wu J, Chen L, et al. A fluorometric assay of SIRT1 deacetylation activity through quantification of nicotinamide adenine dinucleotide. Anal Biochem 2009;395:205-10
71. Sugawara K, Oyama F. Fluorogenic reaction and specific microdetermination of ammonia. J Biochem 1981;89:771-4
72. Garske AL, Denu JM. SIRT1 top 40 hits: use of one-bead, one-compound acetyl-peptide libraries and quantum dots to probe deacetylase specificity. Biochemistry 2006;45:94-101
73. Bheda P, Swatkoski S, Fiedler KL, et al. Biotinylation of lysine method identifies acetylated histone H3 lysine 79 in Saccharomyces cerevisiae as a substrate for Sir2. Proc Natl Acad Sci USA 2012;109:E916-25
74. Park J, Chen Y, Tishkoff DX, et al. SIRT5-mediated lysine desuccinylation impacts diverse metabolic pathways. Mol Cell 2013;50:919-30
75. Nakagawa T, Lomb DJ, Haigis MC, Guarente L. SIRT5 deacetylates carbamoyl phosphate synthetase 1 and regulates the urea cycle. Cell 2009;137:560-70
76. Schlicker C, Gertz M, Papatheodorou P, et al. Substrates and regulation mechanisms for the human mitochondrial Sirtuins Sirt3 and Sirt5. J Mol Biol 2008;382:790-801
77. Zerweck J, Masch A, Schutkowski M. Peptide microarrays for profiling of modification state-specific antibodies. Methods Mol Biol 2009;524:169-80
78. Pacholec M, Bleasdale JE, Chrunyk B, et al. SRT1720, SRT2183, SRT1460, and resveratrol are not direct activators of SIRT1. J Biol Chem 2010;285:8340-51
79. Schlicker C, Boanca G, Lakshminarasimhan M, Steegborn C. Structure-based development of novel Sirtuin inhibitors. Aging (Albany NY) 2011;3:852-7
80. Robers MB, Loh C, Carlson CB, et al. Measurement of the cellular deacetylase activity of SIRT1 on p53 via LanthaScreen(R) technology. Mol Biosyst 2011;7:59-66
81. Dudek JM, Horton RA. TR-FRET biochemical assays for detecting posttranslational modifications of p53. J Biomol Screen 2010;15:569-75
82. Machleidt T, Robers MB, Hermanson SB, et al. TR-FRET cellular assays for interrogating posttranslational modifications of histone H3. J Biomol Screen 2011;16:1236-46
83. Fischer F, Gertz M, Suenkel B, et al. Sirt5 deacylation activities show differential sensitivities to nicotinamide inhibition. PLoS One 2012;7:e45098
84. Rye PT, Frick LE, Ozbal CC, Lamarr WA. Advances in label-free screening approaches for studying sirtuinmediated deacetylation. J Biomol Screen 2011;16:1217-26
85. Bharathi SS, Zhang Y, Mohsen AW, et al. SIRT3 regulates long-chain acyl- CoA dehydrogenase by deacetylating conserved lysines near the active site. J Biol Chem 2013;288(47):33837-47
86. Smith BC, Hallows WC, Denu JM. A continuous microplate assay for sirtuins and nicotinamide-producing enzymes. Anal Biochem 2009;394:101-9
87. Kaeberlein M, McDonagh T, Heltweg B, et al. Substrate-specific activation of sirtuins by resveratrol. J Biol Chem 2005;280:17038-45
88. Madsen AS, Olsen CA. Substrates for efficient fluorometric screening employing the NAD-dependent sirtuin 5 lysine deacylase (KDAC) enzyme. J Med Chem 2012;55:5582-90
89. Hu J, He B, Bhargava S, Lin H. A fluorogenic assay for screening Sirt6 modulators. Org Biomol Chem 2013;11:5213-16
90. Schultz BE, Misialek S, Wu J, et al. Kinetics and comparative reactivity of human class I and class IIb histone deacetylases. Biochemistry 2004;43:11083-91
91. Halley F, Reinshagen J, Ellinger B, et al. A bioluminogenic HDAC activity assay: validation and screening. J Biomol Screen 2011;16:1227-35
92. Pergolizzi G, Butt JN, Bowater RP, Wagner GK. A novel fluorescent probe for NAD-consuming enzymes. Chem Commun (Camb) 2011;47:12655-7
93. Baba R, Hori Y, Mizukami S, Kikuchi K. Development of a fluorogenic probe with a transesterification switch for detection of histone deacetylase activity. J Am Chem Soc 2012;134:14310-13
94. Sankaranarayanapillai M, Tong WP, Yuan Q, et al. Monitoring histone deacetylase inhibition in vivo: noninvasive magnetic resonance spectroscopy method. Mol Imaging 2008;7:92-100
95. Dose A, Liokatis S, Theillet FX, et al. NMR profiling of histone deacetylase and acetyl-transferase activities in real time. ACS Chem Biol 2011;6:419-24
96. Garrity J, Gardner JG, Hawse W, et al. N-lysine propionylation controls the activity of propionyl-CoA synthetase. J Biol Chem 2007;282:30239-45
97. Chen Y, Sprung R, Tang Y, et al. Lysine propionylation and butyrylation are novel post-translational modifications in histones. Mol Cell Proteomics 2007;6:812-19
98. Stevenson FT, Bursten SL, Fanton C, et al. The 31-kDa precursor of interleukin 1 alpha is myristoylated on specific lysines within the 16-kDa Nterminal propiece. Proc Natl Acad Sci USA 1993;90:7245-9
99. Yang B, Zwaans BM, Eckersdorff M, Lombard DB. The sirtuin SIRT6 deacetylates H3 K56Ac in vivo to promote genomic stability. Cell Cycle 2009;8:2662-3
100. Weinert BT, Iesmantavicius V, Wagner SA, et al. Acetyl-phosphate is a critical determinant of lysine acetylation in E. coli. Mol Cell 2013;51:265-72
101. Lammers M, Neumann H, Chin JW, James LC. Acetylation regulates cyclophilin A catalysis, immunosuppression and HIV isomerization. Nat Chem Biol 2010;6:331-7
102. Baur JA, Sinclair DA. Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov 2006;5:493-506
103. Disch JS, Evindar G, Chiu CH, et al. Discovery of thieno[3,2-d] pyrimidine-6- carboxamides as potent inhibitors of SIRT1, SIRT2, and SIRT3. J Med Chem 2013;56:3666-79
104. Hirsch BM, Zheng W. Sirtuin mechanism and inhibition: explored with N(epsilon)-acetyl-lysine analogs. Mol Biosyst 2011;7:16-28
105. Huang R, Holbert MA, Tarrant MK, et al. Site-specific introduction of an acetyl-lysine mimic into peptides and proteins by cysteine alkylation. J Am Chem Soc 2010;132:9986-7
106. Jamonnak N, Fatkins DG, Wei L, Zheng W. N(epsilon)-methanesulfonyllysine as a non-hydrolyzable functional surrogate for N(epsilon)-acetyl-lysine. Org Biomol Chem 2007;5:892-6
107. Asaba T, Suzuki T, Ueda R, et al. Inhibition of human sirtuins by in situ generation of an acetylated lysine-ADP-ribose conjugate. J Am Chem Soc 2009;131:6989-96
108. Smith BC, Denu JM. Mechanism-based inhibition of Sir2 deacetylases by thioacetyl-lysine peptide. Biochemistry 2007;46:14478-86
109. Fatkins DG, Monnot AD, Zheng W. Nepsilon-thioacetyl-lysine: a multi-facet functional probe for enzymatic protein lysine Nepsilon-deacetylation. Bioorg Med Chem Lett 2006;16:3651-6
110. Huhtiniemi T, Salo HS, Suuronen T, et al. Structure-based design of pseudopeptidic inhibitors for SIRT1 and SIRT2. J Med Chem 2011;54:6456-68
111. Jin L, Galonek H, Israelian K, et al. Biochemical characterization, localization, and tissue distribution of the longer form of mouse SIRT3. Protein Sci 2009;18:514-25
112. Nguyen GT, Schaefer S, Gertz M, et al. Structures of human Sirtuin 3 complexes with ADP-ribose and with carba-NAD+ and SRT1720: binding details and inhibition mechanism. Acta Crystallogr D Biol Crystallogr 2013;69:1423-32
113. Moniot S, Schutkowski M, Steegborn C. Crystal structure analysis of human Sirt2 and its ADP-ribose complex. J Struct Biol 2013;182:136-43
114. Borra MT, Smith BC, Denu JM. Mechanism of human SIRT1 activation by resveratrol. J Biol Chem 2005;280:17187-95
115. Lakshminarasimhan M, Curth U, Moniot S, et al. Molecular architecture of the human protein deacetylase Sirt1 and its regulation by AROS and resveratrol. Biosci Rep 2013;33:e00037
116. Nguyen GT, Gertz M, Steegborn C. Crystal structures of Sirt3 complexes with 4'-bromo-resveratrol reveal binding sites and inhibition mechanism. Chem Biol 2013;20:1375-85
117. Baur JA, Chen D, Chini EN, et al. Dietary restriction: standing up for Sirtuins. Science 2010;329:1012-13; author reply 13-4
118. Szczepankiewicz BG, Dai H, Koppetsch KJ, et al. Synthesis of carba- NAD and the structures of its ternary complexes with SIRT3 and SIRT5. J Org Chem 2012;77:7319-29