US 2009.0143376A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2009/0143376 A1 Milburn et al. (43) Pub. Date: Jun. 4, 2009

(54) FUSED HETEROCYCLIC COMPOUNDS AND A6II 3/4439 (2006.01) THEIR USE ASSRTUIN MODULATORS C07D 277/62 (2006.01) C07D 239/72 (2006.01) (75) Inventors: Michael Milburn, Cary, NC (US); C07D 209/04 (2006.01) Jill Milne, Brookline, MA (US); A6IP 2L/00 (2006.01) Jean Bemis, Arlington, MA (US); A6IP 7/00 (2006.01) Joseph J. Nunes, Andover, MA A6IP 25/00 (2006.01) (US); Roger Xie, Southborough, A6IP3/00 (2006.01) MA (US); Karl D. Nomington, A63/496 (2006.01) Acton, MA (US); Chi B. Vu, C07D 40/4 (2006.01) Arlington, MA (US) C07D 413/14 (2006.01) A63L/437 (2006.01) Correspondence Address: C07D 471/02 (2006.01) ROPES & GRAY LLP A6IP35/00 (2006.01) PATENT DOCKETING 39/41, ONE INTERNA- A 6LX 3/57 (2006.01) TIONAL PLACE A6II 3/428 (2006.01) BOSTON, MA 02110-2624 (US) C07D 23L/56 (2006.01) A63L/404 (2006.01) (73) Assignee: Sitris Pharmaceuticals, Inc., C07D 513/02 (2006.01) Cambridge, MA (US) A6II 3/429 (2006.01) (21) Appl. No.: 11/885,576 C07D 417/02 (2006.01) (52) U.S. Cl...... 514/233.2: 548/305.1: 514/394; (22) PCT Filed: Mar. 3, 2006 435/375; 435/377: 548/305.4; 546/273.4: 514/338: 548/180: 514/367: 548/362. 1: 514/.407; (86). PCT No.: PCT/US06/07744 548/151; 514/366; 54.4/135; 514/233.8:544/284; 514/266.2: 548/465; 514/.414: 546/121; 514/300; S371 (c)(1), 544/127: 544/362; 514/253.04 (2), (4) Date: Mar. 4, 2008 (57) ABSTRACT Related U.S. Application Data Provided herein are novel sirtuin-modulating compounds and (60) Provisional application No. 60/658.430, filed on Mar. methods of use thereof. The sirtuin-modulating compounds 3, 2005, provisional application No. 60/705.386, filed may be used for increasing the lifespan of a cell, and treating on Aug. 4, 2005. and/or preventing a wide variety of diseases and disorders including, for example, diseases or disorders related to aging Publication Classification or stress, diabetes, obesity, neurodegenerative diseases, che (51) Int. Cl. motherapeutic induced neuropathy, neuropathy associated A 6LX 3/5.377 (2006.01) with an ischemic event, polyglutamine diseases, ocular dis CO7D 235/04 (2006.01) eases and/or disorders, cardiovascular disease, blood clotting A6 IK 3/484 (2006.01) disorders, inflammation, cancer, and/or flushing. Also pro CI2N 5/00 (2006.01) vided are compositions comprising a sirtuin-modulating CO7D 40/02 (2006.01) compound in combination with another therapeutic agent. US 2009/0143376 A1 Jun. 4, 2009

FUSED HETEROCYCLIC COMPOUNDS AND located at the N-terminus. SIRT3 has NAD+-dependent pro THEIR USE ASSRTUIN MODULATORS tein deacetylase activity and is ubiquitously expressed, par ticularly in metabolically active tissues. Upon transfer to the RELATED APPLICATIONS mitochondria, SIRT3 is believed to be cleaved into a smaller, 0001. This application claims the benefit of priority to active form by a mitochondrial matrix processing peptidase U.S. Provisional Application Nos. 60/658.430, filed Mar. 3, (MPP) (B. Schwer et al. J. Cell Biol. 158: 647-657 (2002)). 2005, and 60/705.386, filed Aug. 4, 2005, which applications 0007 Caloric restriction has been known for over 70 years are hereby incorporated by reference in their entireties. to improve the health and extend the lifespan of mammals (Masoro, 2000). Yeast lifespan, like that of metazoans, is also BACKGROUND extended by interventions that resemble caloric restriction, such as low glucose. The discovery that both yeast and flies 0002. The Silent Information Regulator (SIR) family of genes represents a highly conserved group of genes present in lacking the SIR2 gene do not live longer when calorically the genomes of organisms ranging from archaebacteria to a restricted provides evidence that SIR2 genes mediate the variety of eukaryotes (Frye, 2000). The encoded SIR proteins beneficial health effects of this diet (Anderson et al., 2003: are involved in diverse processes from regulation of gene Helfand and Rogina, 2004). Moreover, mutations that reduce silencing to DNA repair. The proteins encoded by members of the activity of the yeast glucose-responsive cAMP (adenosine the SIR gene family show high sequence conservation in a 3'5'-monophosphate)-dependent (PKA) pathway extend life 250 amino acid core domain. A well-characterized gene in span in wild type cells but not in mutant sir2 Strains, demon this family is S. cerevisiae SIR2, which is involved in silenc strating that SIR2 is likely to be a key downstream component ing HM loci that contain information specifying yeast mating of the caloric restriction pathway (Lin et al., 2001). type, telomere position effects and cell aging (Guarente, 1999: Kaeberlein et al., 1999: Shore, 2000). The yeast Sir2 SUMMARY protein belongs to a family of histone deacetylases (reviewed in Guarente, 2000; Shore, 2000). The Sir2 homolog, CobB, in 0008 Provided herein are novel sirtuin-modulating com Salmonella typhimurium, functions as an NAD (nicotinamide pounds and methods of use thereof. adenine dinucleotide)-dependent ADP-ribosyl transferase 0009. In one aspect, the invention provides novel sirtuin (Tsang and Escalante-Semerena, 1998). modulating compounds of Formula (I): 0003. The Sir2 protein is a class III deacetylase which uses NAD as a cosubstrate (Imai et al., 2000; Moazed, 2001; Smith et al., 2000; Tanner et al., 2000; Tanny and Moazed, 2001). (I) Unlike other deacetylases, many of which are involved in gene silencing, Sir2 is insensitive to class I and II histone deacetylase inhibitors like trichostatin A (TSA) (Imai et al., 2000; Landry et al., 2000a: Smith et al., 2000). 0004 Deacetylation of acetyl-lysine by Sir2 is tightly coupled to NAD hydrolysis, producing nicotinamide and a novel acetyl-ADP ribose compound (Tanner et al., 2000; Landry et al., 2000b: Tanny and Moazed, 2001). The NAD dependent deacetylase activity of Sir2 is essential for its functions which can connect its biological role with cellular or a salt thereof, where, as Valence permits: metabolism in yeast (Guarente, 2000: Imai et al., 2000; Linet 0010 Ring A is optionally Substituted; al., 2000; Smith et al., 2000). Mammalian Sir2 homologs 0011 L is absent, substituted or unsubstituted phenylene, have NAD-dependent histone deacetylase activity (Imai et substituted or unsubstituted —O-phenylene, substituted or al., 2000; Smith et al., 2000). Most information about Sir2 unsubstituted thienylene, substituted or unsubstituted pyra mediated functions comes from the studies in yeast (Garten Zolylene, substituted or unsubstituted benzothiazolylene, berg, 2000: Gottschling, 2000). 0005 Biochemical studies have shown that Sir2 can NR, C(O) NRs , S—, CHR =CHR, or readily deacetylate the amino-terminal tails of histones H3 —CHR C(O)—: and H4, resulting in the formation of 1-O-acetyl-ADP-ribose 0012 L' is absent, substituted or unsubstituted phenylene, and nicotinamide. Strains with additional copies of SIR2 substituted or unsubstituted —O-phenylene, substituted or display increased rDNA silencing and a 30% longer lifespan. unsubstituted thienylene, substituted or unsubstituted pyra It has recently been shown that additional copies of the C. Zolylene, substituted or unsubstituted benzothiazolylene, elegans SIR2 homolog, Sir-2.1, and the D. melanogaster substituted or unsubstituted indenedionylene. —C(O)O—, dSir2 gene greatly extend life span in those organisms. This —C(O)NR —NRC(O)— —NR C(O) NRs , implies that the SIR2-dependent regulatory pathway for —S—, —CHR=CHR, or —CHRs C(O)—, provided aging arose early in evolution and has been well conserved. that at least one of L and L' is substituted or unsubstituted Today, Sir2 genes are believed to have evolved to enhance an phenylene, Substituted or unsubstituted—O-phenylene, Sub organism's health and stress resistance to increase its chance stituted or unsubstituted thienylene, substituted or unsubsti of Surviving adversity. tuted pyrazolylene, substituted or unsubstituted benzothiaz 0006 SIRT3 is a homolog of SIRT1 that is conserved in olylene, substituted or unsubstituted indenedionylene, prokaryotes and eukaryotes (P. Onyango et al., Proc. Natl. NR , C(O)O , C(O)NR , NRC(O) , Acad. Sci. USA99: 13653-13658 (2002)). The SIRT3 protein NR C(O) NRs , S—, CHR =CHR or is targeted to the mitochondrial cristae by a unique domain US 2009/0143376 A1 Jun. 4, 2009

0013 R is absent, —H, NRRs —NC(O)Rs —ORs, —S—(C-C straight or branched alkyl), —N(Rao)(Rs), naphthyl or a heterocyclic group, provided that L and R are —S(O), N(Ra)(Rs), heterocyclyl (C-C straight or branched alkyl)-heterocyclyl. —O—(C-C straight or not both absent unless X is N: branched alkyl)-heterocyclyl. —S (C-C straight or 0014 R is —H, unsubstituted alkyl, - NRRs, NRC branched alkyl)-heterocyclyl, or is optionally fused to a 5-6 (O)Rs —ORs, substituted or unsubstituted phenyl, naphthyl membered heterocyclyl or heteroaryl, wherein any heterocy or a heterocyclic group; clyl or heteroaryl is optionally substituted with —C-C, 0015 R is —H, NRRs, NC(O)Rs —ORs or a sub straight or branched alkyl. stituted or unsubstituted heterocyclic group, or R and Rs. 0027. In a further aspect, the invention provides novel taken together with the atoms to which they are attached, form sirtuin-modulating compounds of Formula (Ib): an optionally Substituted heterocyclic group, or R is absent (Ib) when Z is O or S: 0016 Ra and Rs are independently —H, a substituted or unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic hetero cyclic group: R11 0017 R. R., and R are independently selected from the V group consisting of halogen, —OR, —CN, —COR, NH —OCOR —OCOR —C(O)NRRs —OC(O)NRRs, R —C(O)R. —COR - SR —OSOH, -S(O), R = S(O) 21 N 3-k 2 OR —S(O)NRRs —NRRs —NRC(O)ORs, —NRC S. N (O)Rs and—NO; N N N R13, 0018 W is C or N: 0019 X is C or N: R10 0020 Y is C or N: wherein: 0021 Z is C, N, O or S, provided that at least two of W, X, 0028 Z is selected from O or S: Y and Z are C; and 0029 Rio is selected from —H, C(O) N(R)(Rs), —S(O)N(Rao)(Rs), or —CH2—N(Rao)(Rs); wherein 0022 n is 1 or 2. each of Rao and Rso is independently selected from —H. 0023. In a second aspect, the invention provides novel —C-C straight or branched alkyl, —(C-C straight or sirtuin-modulating compounds represented by Structural branched alkyl)-N(CH), —(C-C straight or branched Formula (Ia): alkyl)-heterocyclyl. —(C-C straight or branched alkyl)- alkylheterocyclyl, or wherein Rao and Rso taken together with the Natom to which they are bound form a 5-6 membered (Ia) heterocyclic ring that is optionally substituted with —(C-C, straight or branched alkyl), and wherein at least one of Rao or Rs is not H. 0030 R is selected from —C-C straight or branched alkylene or —C(O)—: 0031 each of R and R is independently selected from R11 —H or —(C-C straight or branched alkyl), or R and R. V are taken together to form a benzene ring that is Substituted NH with up to two substituents independently selected from —(C-C straight or branched alkyl). —CF or halo; and 21 NN N 0032 ring K is substituted with up to three substituents independently selected from halo. —CF, —O—(C-C, N1sN straight or branched alkyl). —S (C-C straight or branched alkyl). —N(R)(Rs). —S(O), N(R)(Rs), heterocyclyl, R10 (C-C straight or branched alkyl)-heterocyclyl. —O—(C- C straight or branched alkyl)-heterocyclyl. —S-(C-C, wherein: straight or branched alkyl)-heterocyclyl, or is optionally fused to a 5-6 membered heterocyclyl or heteroaryl, wherein 0024 Rio is selected from —H. —C(O)—N(R)(Rs), any heterocyclyl or heteroaryl is optionally substituted with —S(O)N(Rao)(Rs), or —CH2—N(Rao)(Rs); wherein —C-C straight or branched alkyl. each of Rao and Rso is independently selected from —H. 0033. In another aspect, the invention provides novel sir —C-C straight or branched alkyl, —(C-C straight or tuin-modulating compounds of Formula (II): branched alkyl)-N(CH), —(C-C straight or branched alkyl)-heterocyclyl. —(C-C straight or branched alkyl)- alkylheterocyclyl, or wherein Rao and Rso taken together with (II) the Natom to which they are bound form a 5-6 membered heterocyclic ring that is optionally substituted with —(C-C, straight or branched alkyl), and wherein at least one of Rao or Rso is not H; 0025 R, is selected from C-C straight or branched alkylene or —C(O)—, and 0026 each of ring K and ring E is independently substi or a salt thereof, where: tuted with up to three substituents independently selected 0034 Rings C, D and E are optionally substituted; and from halo. —CF, —O—(C-C straight or branched allyl), 0035) x is 0 or 1. US 2009/0143376 A1 Jun. 4, 2009

0036. In yet another aspect, the invention provides novel described further below, the methods comprise administering sirtuin-modulating compounds of Formula (VII): to a subject in need thereof a pharmaceutically effective amount of a sirtuin-modulating compound. 0048. In certain aspects, the sirtuin-modulating com (VII) R3 pounds may be administered alone or in combination with M other compounds, including other sirtuin-modulating com Z R2, pounds, or other therapeutic agents. y M s L DETAILED DESCRIPTION 1. Definitions or a salt thereof, where: 0049. As used herein, the following terms and phrases 0037 Ring F is optionally substituted; shall have the meanings set forth below. Unless defined oth 0038 L' is substituted or unsubstituted phenylene, substi erwise, all technical and scientific terms used herein have the tuted or unsubstituted thienylene. —C(O)O— —S same meaning as commonly understood to one of ordinary —CHR=CHR or —CHRs C(O)—: skill in the art. 0039 R is —H, unsubstituted alkyl, - NRRs —NRC 0050. The singular forms “a,” “an and “the include plu (O)Rs, —ORs or a heterocyclic group; ral reference unless the context clearly dictates otherwise. 0040 R is H. —NRRs, NC(O)Rs—ORs or a het 0051. The term "agent' is used hereinto denote a chemical erocyclic group, or R and R, taken together with the atoms compound, a mixture of chemical compounds, a biological to which they are attached, form an optionally substituted macromolecule (such as a nucleic acid, an antibody, a protein heterocyclic group, or R is absent when Z is O or S; or portion thereof, e.g., a peptide), or an extract made from 0041 Ra and Rs are independently —H, a substituted or biological materials such as bacteria, plants, fungi, or animal unsubstituted alkyl group, a Substituted or unsubstituted aryl (particularly mammalian) cells or tissues. The activity of Such group or a Substituted or unsubstituted non-aromatic hetero agents may render it Suitable as a “therapeutic agent' which is cyclic group: a biologically, physiologically, or pharmacologically active 0042. R. R., and Rs are independently selected from the Substance (or Substances) that acts locally or systemically in group consisting of halogen, —OR, —CN, —COR, a Subject. —OCOR —OCOR = C(O)NRRs —OC(O)NRRs, 0052. The term “bioavailable” when referring to a com —C(O)R. —COR —SR —OSOH, -S(O), R —S(O) pound is art-recognized and refers to a form of a compound OR - S(O)NRRs, NRRs, NRC(O)ORs, NRC that allows for it, or a portion of the amount of compound (O)Rs and—NO; administered, to be absorbed by, incorporated to, or otherwise 0043 Z is C, N, O or S; and physiologically available to a subject or patient to whom it is 0044 n is 1 or 2. administered. 0045. The invention also includes salts, prodrugs and 0053 “Biologically active portion of a sirtuin” refers to a metabolites of the compounds disclosed herein. portion of a sirtuin protein having a biological activity, Such 0046. Also provided are pharmaceutical compositions as the ability to deacetylate. Biologically active portions of a comprising one or more compounds of Formulas (I)-(X) or a sirtuin may comprise the core domain of a sirtuin. Biologi salt, prodrug, or metabolite thereof. cally active portions of SIRT1 having GenBank Accession 0047. In another aspect, the invention provides methods No. NP 036370 that encompass the NAD+ binding domain for using sirtuin-modulating compounds, or compostions and the Substrate binding domain, for example, may include comprising sirtuin-modulating compounds. In certain without limitation, amino acids 62-293 of GenBank Acces embodiments, sirtuin-modulating compounds that increase sion No. NP 036370, which are encoded by nucleotides 237 the level and/or activity of a sirtuin protein may be used for a to 932 of GenBank Accession No. NM 012238. Therefore, variety of therapeutic applications including, for example, this region is sometimes referred to as the core domain. increasing the lifespan of a cell, and treating and/or prevent 0054) Other biologically active portions of SIRT1, also ing a wide variety of diseases and disorders including, for Sometimes referred to as core domains, include about amino example, diseases or disorders related to aging or stress, acids 261 to 447 of GenBank Accession No. NP 036370, diabetes, obesity, neurodegenerative diseases, chemothera which are encoded by nucleotides 834 to 1394 of GenBank peutic induced neuropathy, neuropathy associated with an Accession No. NM 012238; aboutamino acids 242 to 493 of ischemic event, ocular diseases and/or disorders, cardiovas GenBank Accession No. N 036370, which are encoded by cular disease, blood clotting disorders, inflammation, and/or nucleotides 777 to 1532 of GenBank Accession No. flushing, etc. Sirtuin-modulating compounds that increase NM 012238; or about amino acids 254 to 495 of GenBank the level and/or activity of a sirtuin protein may also be used Accession No. NP 036370, which are encoded by nucle for treating a disease or disorder in a Subject that would otides 813 to 1538 of GenBank Accession No. NM 012238. benefit from increased mitochondrial activity, for enhancing 0055. The term “companion animals' refers to cats and muscle performance, for increasing muscle ATP levels, or for dogs. As used herein, the term “dog(s) denotes any member treating or preventing muscle tissue damage associated with of the species Canis familiaris, of which there are a large hypoxia or ischemia. In other embodiments, sirtuin-modulat number of different breeds. The term “cat(s) refers to a feline ing compounds that decrease the level and/or activity of a animal including domestic cats and other members of the sirtuin protein may be used for a variety of therapeutic appli family Felidae, genus Felis. cations including, for example, increasing cellular sensitivity 0056. The terms “comprise' and “comprising are used in to stress, increasing apoptosis, treatment of cancer, Stimula the inclusive, open sense, meaning that additional elements tion of appetite, and/or stimulation of weight gain, etc. As may be included. US 2009/0143376 A1 Jun. 4, 2009

0057 The term “conserved residue' refers to an amino contributed to by insulin resistance. Examples include: dia acid that is a member of a group of amino acids having certain betes, obesity, metabolic syndrome, insulin-resistance Syn common properties. The term "conservative amino acid Sub dromes, syndrome X, insulin resistance, high blood pressure, stitution” refers to the substitution (conceptually or other hypertension, high blood cholesterol, dyslipidemia, hyper wise) of an amino acid from one such group with a different lipidemia, dyslipidemia, atherosclerotic disease including amino acid from the same group. A functional way to define stroke, coronary artery disease or myocardial infarction, common properties between individual amino acids is to hyperglycemia, hyperinsulinemia and/or hyperproinsuline analyze the normalized frequencies of amino acid changes mia, impaired glucose tolerance, delayed insulin release, dia between corresponding proteins of homologous organisms betic complications, including coronary heart disease, angina (Schulz, G. E. and R. H. Schirmer. Principles of Protein pectoris, congestive heart failure, stroke, cognitive functions Structure, Springer-Verlag). According to such analyses, in dementia, retinopathy, peripheral neuropathy, nephropa groups of amino acids may be defined where amino acids thy, glomerulonephritis, glomerulosclerosis, nephrotic Syn within a group exchange preferentially with each other, and drome, hypertensive nephrosclerosis some types of cancer therefore resemble each other most in their impact on the (such as endometrial, breast prostate, and colon), complica overall protein structure (Schulz, G. E. and R. H. Schirmer, tions of pregnancy, poor female reproductive health (Such as Principles of Protein Structure, Springer-Verlag). One menstrual irregularities, infertility, irregular ovulation, poly example of a set of amino acid groups defined in this manner cystic ovarian syndrome (PCOS)), lipodystrophy, cholesterol include: (i) a charged group, consisting of Glu and Asp, LyS, related disorders, such as gallstones, cholescystitis and Arg and His, (ii) a positively-charged group, consisting of cholelithiasis, gout, obstructive sleep apnea and respiratory Lys, Arg and His, (iii) a negatively-charged group, consisting problems, osteoarthritis, and prevention and treatment of of Glu and Asp, (iv) an aromatic group, consisting of Phe, Tyr bone loss, e.g. osteoporosis. and Trp, (v) a nitrogen ring group, consisting of His and Trp, 0065. The term “livestock animals' refers to domesticated (vi) a large aliphatic nonpolar group, consisting of Val, Leu quadrupeds, which includes those being raised for meat and and Ile, (vii) a slightly-polar group, consisting of Met and various byproducts, e.g., a bovine animal including cattle and Cys, (viii) a Small-residue group, consisting of Ser, Thr, Asp, other members of the genus Bos, a porcine animal including ASn, Gly, Ala, Glu, Gln and Pro, (ix) an aliphatic group domestic Swine and other members of the genus Sus, an ovine consisting of Val, Leu, Ile, Met and Cys, and (X) a small animal including sheep and other members of the genus Ovis, hydroxyl group consisting of Ser and Thr. domestic goats and other members of the genus Capra. 0058 “Diabetes’ refers to high blood sugar or ketoacido domesticated quadrupeds being raised for specialized tasks sis, as well as chronic, general metabolic abnormalities aris Such as use as a beast of burden, e.g., an equine animal ing from a prolonged high blood Sugar status or a decrease in including domestic horses and other members of the family glucose tolerance. “Diabetes’ encompasses both the type I Equidae, genus Equus. and type II (Non Insulin Dependent Diabetes Mellitus or 0066. The term “mammal’ is known in the art, and exem NIDDM) forms of the disease. The risk factors for diabetes plary mammals include humans, primates, livestock animals include the following factors: waistline of more than 40 (including bovines, porcines, etc.), companion animals (e.g., inches for men or 35 inches for women, blood pressure of canines, felines, etc.) and rodents (e.g., mice and rats). 130/85 mmHg or higher, triglycerides above 150 mg/dl., fast 0067. The term “naturally occurring form' when referring ing blood glucose greater than 100 mg/dl or high-density to a compound means a compound that is in a form, e.g., a lipoprotein of less than 40 mg/dl in men or 50 mg/dl in composition, in which it can be found naturally. For example, WOC. since resveratrol can be found in red wine, it is present in red 0059. A “direct activator of a sirtuin is a molecule that wine in a form that is naturally occurring. A compound is not activates a sirtuin by binding to it. A “direct inhibitor of a in a form that is naturally occurring if, e.g., the compound has sirtuin is a molecule inhibits a sirtuin by binding to it. been purified and separated from at least some of the other 0060. The term “EDs is art-recognized. In certain molecules that are found with the compound in nature. A embodiments, EDso means the dose of a drug which produces “naturally occurring compound” refers to a compound that 50% of its maximum response or effect, or alternatively, the can be found in nature, i.e., a compound that has not been dose which produces a pre-determined response in 50% of designed by man. A naturally occurring compound may have test Subjects or preparations. The term “LDso is art-recog been made by man or by nature. nized. In certain embodiments, LDso means the dose of a drug 0068 A “naturally occurring compound” refers to a com which is lethal in 50% of test subjects. The term “therapeutic pound that can be found in nature, i.e., a compound that has index' is an art-recognized term which refers to the therapeu not been designed by man. A naturally occurring compound tic index of a drug, defined as LDs/EDs. may have been made by man or by nature. For example, 0061 The term “hyperinsulinemia' refers to a state in an resveratrol is a naturally-occurring compound. A “non-natu individual in which the level of insulin in the blood is higher rally occurring compound is a compound that is not known than normal. to exist in nature or that does not occur in nature. 0062. The term “including is used to mean “including but 0069 “Obese" individuals or individuals suffering from not limited to”. “Including and “including but not limited to obesity are generally individuals having a body mass index are used interchangeably. (BMI) of at least 25 or greater. Obesity may or may not be 0063. The term “insulin resistance” refers to a state in associated with insulin resistance. which a normal amount of insulin produces a Subnormal 0070 The terms “parenteral administration' and “admin biologic response relative to the biological response in a istered parenterally’ are art-recognized and refer to modes of Subject that does not have insulin resistance. administration other than enteral and topical administration, 0064. An “insulin resistance disorder,” as discussed usually by injection, and includes, without limitation, intra herein, refers to any disease or condition that is caused by or venous, intramuscular, intraarterial, intrathecal, intracapsu US 2009/0143376 A1 Jun. 4, 2009

lar, intraorbital, intracardiac, intradermal, intraperitoneal, boxymethyl cellulose, ethyl cellulose and cellulose acetate; transtracheal. Subcutaneous, Subcuticular, intra-articulare, (4) powdered tragacanth; (5) malt, (6) gelatin; (7) talc.; (8) Subcapsular, Subarachnoid, intraspinal, and intrasternal injec excipients, such as cocoa butter and Suppository waxes; (9) tion and infusion. oils, such as peanut oil, cottonseed oil, safflower oil, Sesame (0071. A “patient”, “subject”, “individual” or “host” refers oil, olive oil, corn oil and soybean oil: (10) glycols, such as to either a human or a nonhuman animal. propylene glycol; (11) polyols, such as glycerin, Sorbitol, 0072 The term “percent identical refers to sequence mannitol and polyethylene glycol; (12) esters, such as ethyl identity between two amino acid sequences or between two oleate and ethyl laurate; (13) agar, (14) buffering agents. Such nucleotide sequences. Identity can each be determined by as magnesium hydroxide and aluminum hydroxide; (15) alg comparing a position in each sequence which may be aligned inic acid; (16) pyrogen-free water; (17) isotonic saline; (18) for purposes of comparison. When an equivalent position in Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer the compared sequences is occupied by the same base or Solutions; and (21) other non-toxic compatible Substances amino acid, then the molecules are identical at that position; employed in pharmaceutical formulations. when the equivalent site occupied by the same or a similar amino acid residue (e.g., similar in Steric and/or electronic (0075. The terms “polynucleotide', and “nucleic acid” are nature), then the molecules can be referred to as homologous used interchangeably. They refer to a polymeric form of (similar) at that position. Expression as a percentage of nucleotides of any length, either deoxyribonucleotides or homology, similarity, or identity refers to a function of the ribonucleotides, or analogs thereof. Polynucleotides may number of identical or similar amino acids at positions shared have any three-dimensional structure, and may perform any by the compared sequences. Expression as a percentage of function, known or unknown. The following are non-limiting homology, similarity, or identity refers to a function of the examples of polynucleotides: coding or non-coding regions number of identical or similar amino acids at positions shared of a gene or gene fragment, loci (locus) defined from linkage by the compared sequences. Various alignment algorithms analysis, exons, introns, messenger RNA (mRNA), transfer and/or programs may be used, including FASTA, BLAST, or RNA, ribosomal RNA, ribozymes, cDNA, recombinant poly ENTREZ. FASTA and BLAST are available as a part of the nucleotides, branched polynucleotides, plasmids, vectors, GCG sequence analysis package (University of Wisconsin, isolated DNA of any sequence, isolated RNA of any Madison, Wis.), and can be used with, e.g., default settings. sequence, nucleic acid probes, and primers. A polynucleotide ENTREZ is available through the National Center for Bio may comprise modified nucleotides, such as methylated technology Information, National Library of Medicine, nucleotides and nucleotide analogs. If present, modifications National Institutes of Health, Bethesda, Md. In one embodi to the nucleotide structure may be imparted before or after ment, the percent identity of two sequences can be deter assembly of the polymer. The sequence of nucleotides may be mined by the GCG program with a gap weight of 1, e.g., each interrupted by non-nucleotide components. A polynucleotide amino acid gap is weighted as if it were a singleamino acid or may be further modified, such as by conjugation with a label nucleotide mismatch between the two sequences. ing component. The term “recombinant polynucleotide 0073. Other techniques for alignment are described in means a polynucleotide of genomic, cDNA, semisynthetic, or Methods in Enzymology, Vol. 266: Computer Methods for Macromolecular Sequence Analysis (1996), ed. Doolittle, synthetic origin which either does not occur in nature or is Academic Press, Inc., a division of Harcourt Brace & Co., San linked to another polynucleotide in a nonnatural arrange Diego, Calif., USA. Preferably, an alignment program that ment. permits gaps in the sequence is utilized to align the sequences. (0076. The term “prophylactic' or “therapeutic” treatment The Smith-Waterman is one type of algorithm that permits is art-recognized and refers to administration of a drug to a gaps in sequence alignments. See Meth. Mol. Biol. 70: 173 host. If it is administered prior to clinical manifestation of the 187 (1997). Also, the GAP program using the Needleman and unwanted condition (e.g., disease or other unwanted State of Wunsch alignment method can be utilized to align sequences. the host animal) then the treatment is prophylactic, i.e., it An alternative search strategy uses MPSRCH software, protects the host against developing the unwanted condition, which runs on a MASPAR computer. MPSRCH uses a Smith whereas if administered after manifestation of the unwanted Waterman algorithm to score sequences on a massively par condition, the treatment is therapeutic (i.e., it is intended to allel computer. This approach improves ability to pick up diminish, ameliorate or maintain the existing unwanted con distantly related matches, and is especially tolerant of Small dition or side effects therefrom). gaps and nucleotide sequence errors. Nucleic acid-encoded 0077. The term “protecting group' is art-recognized and amino acid sequences can be used to search both protein and refers to temporary Substituents that protect a potentially DNA databases. reactive functional group from undesired chemical transfor 0074 The term “pharmaceutically acceptable carrier is mations. Examples of Such protecting groups include esters art-recognized and refers to a pharmaceutically-acceptable of carboxylic acids, silyl ethers of alcohols, and acetals and material, composition or vehicle. Such as a liquid or Solid ketals of aldehydes and ketones, respectively. The field of filler, diluent, excipient, Solvent or encapsulating material, protecting group chemistry has been reviewed by Greene and involved in carrying or transporting any Subject composition Wuts in Protective Groups in Organic Synthesis (2" ed., or component thereof. Each carrier must be “acceptable' in Wiley: New York, 1991). the sense of being compatible with the Subject composition 0078. The term "pyrogen-free', with reference to a com and its components and not injurious to the patient. Some position, refers to a composition that does not contain a pyro examples of materials which may serve as pharmaceutically gen in an amount that would lead to an adverse effect (e.g., acceptable carriers include: (1) Sugars, such as lactose, glu irritation, fever, inflammation, diarrhea, respiratory distress, cose and Sucrose; (2) starches, such as corn starch and potato endotoxic shock, etc.) in a Subject to which the composition starch; (3) cellulose, and its derivatives, such as sodium car has been administered. For example, the term is meant to US 2009/0143376 A1 Jun. 4, 2009

encompass compositions that are free of, or substantially free hSIRT4, hSIRT5, hSIRT6 and hSIRT7 (Brachmann et al. of an endotoxin Such as, for example, a lipopolysaccharide (1995) Genes Dev. 9:2888 and Frye et al. (1999) BBRC (LPS). 260:273). Preferred sirtuins are those that share more simi 0079) “Replicative lifespan” of a cell refers to the number larities with SIRT1, i.e., hSIRT1, and/or Sir2 than with of daughter cells produced by an individual “mother cell.” SIRT2, such as those members having at least part of the "Chronological aging or "chronological lifespan on the N-terminal sequence present in SIRT1 and absent in SIRT2 other hand, refers to the length of time a population of non such as SIRT3 has. dividing cells remains viable when deprived of nutrients. I0084) “SIRT1 protein” refers to a member of the sir2 fam “Increasing the lifespan of a cell' or “extending the lifespan ily of sirtuin deacetylases. In one embodiment, a SIRT1 pro of a cell as applied to cells or organisms, refers to increasing tein includes yeast Sir2 (GenBankAccession No. P53685), C. the number of daughter cells produced by one cell; increasing elegans Sir-2.1 (GenBank Accession No. NP 501912), the ability of cells or organisms to cope with stresses and human SIRT1 (GenBank Accession No. NM 012238 or combat damage, e.g., to DNA, proteins; and/or increasing the NP 036370 (or AF083106)), and human SIRT2 (GenBank ability of cells or organisms to Survive and exist in a living Accession No. NM 012237, NM 030593, NP 036369, state for longer under a particular condition, e.g., stress (for NP 085096, or AF083107) proteins, and equivalents and example, heatshock, osmotic stress, high energy radiation, fragments thereof. In another embodiment, a SIRT1 protein chemically-induced stress, DNA damage, inadequate salt includes a polypeptide comprising a sequence consisting of level, inadequate nitrogen level, or inadequate nutrient level). or consisting essentially of the amino acid sequence set forth Lifespan can be increased by at least about 20%, 30%, 40%, in GenBank Accession Nos. NP 036370, NP 501912, 50%, 60% or between 20% and 70%, 30% and 60%, 40% and NP 085096, NP 036369, or P53685. SIRT1 proteins 60% or more using methods described herein. include polypeptides comprising all or a portion of the amino 0080 “Sirtuin-activating compound” refers to a com acid sequence set forth in GenBank Accession Nos. pound that increases the level of a sirtuin protein and/or NP 036370, NP 501912, NP 085096, NP 036369, or increases at least one activity of a sirtuin protein. In an exem P53685; the amino acid sequence set forth in GenBank plary embodiment, a sirtuin-activating compound may Accession Nos. NP 036370, NP 501912, NP 085096, increase at least one biological activity of a sirtuin protein by NP 036369, or P53685 with 1 to about 2, 3, 5, 7, 10, 15, 20, at least about 10%, 25%, 50%, 75%, 100%, or more. Exem 30, 50, 75 or more conservative amino acid substitutions; an plary biological activities of sirtuin proteins include deacety amino acid sequence that is at least 60%, 70%, 80%, 90%, lation, e.g., of histones and p53; extending lifespan; increas 95%, 96%, 97%, 98%, or 99% identical to GenBank Acces ing genomic stability; silencing transcription; and controlling sion Nos. NP 036370, NP 501912, NP 085096, the segregation of oxidized proteins between mother and NP 036369, or P53685, and functional fragments thereof. daughter cells. Polypeptides of the invention also include homologs (e.g., 0081. “Sirtuin-inhibiting compound” refers to a com orthologs and paralogs), variants, or fragments, of GenBank pound that decreases the level of a sirtuin protein and/or Accession Nos. NP 036370, NP 501912, NP 085096, decreases at least one activity of a sirtuin protein. In an exem NP 036369, or P53685. plary embodiment, a sirtuin-inhibiting compound may I0085 “SIRT3 protein” refers to a member of the sirtuin decrease at least one biological activity of a sirtuin protein by deacetylase protein family and/or to a homolog of a SIRT1 at least about 10%, 25%, 50%, 75%, 100%, or more. Exem protein. In one embodiment, a SIRT3 protein includes human plary biological activities of sirtuin proteins include deacety SIRT3 (GenBank Accession No. AAHO1042, NP 036371, lation, e.g., of histones and p53; extending lifespan; increas or NP 001017524) and mouse SIRT3 (GenBank Accession ing genomic stability; silencing transcription; and controlling No. NP 071878) proteins, and equivalents and fragments the segregation of oxidized proteins between mother and thereof. In another embodiment, a SIRT3 protein includes a daughter cells. polypeptide comprising a sequence consisting of, or consist 0082 "Sirtuin-modulating compound” refers to a com ing essentially of the amino acid sequence set forth in Gen pound of Formulas (I)-(X) as described herein. In exemplary Bank Accession Nos. AAHO1042, NP 036371, embodiments, a sirtuin-modulating compound may either up NP 001017524, or NP 071878. SIRT3 proteins include regulate (e.g., activate or stimulate), down regulate (e.g., polypeptides comprising all or a portion of the amino acid inhibit or Suppress) or otherwise change a functional property sequence set forth in GenBank Accession AAHO1042, or biological activity of a sirtuin protein. Sirtuin-modulating NP 036371, NP 001017524, or NP 071878; the amino compounds may act to modulate a sirtuin protein either acid sequence set forth in GenBank Accession Nos. directly or indirectly. In certain embodiments, a sirtuin AAHO1042, NP 036371, NP 001017524, or NP 071878 modulating compound may be a sirtuin-activating compound with 1 to about 2, 3, 5, 7, 10, 15, 20, 30, 50, 75 or more or a sirtuin-inhibiting compound. conservative amino acid Substitutions; an amino acid I0083 “Sirtuin protein” refers to a member of the sirtuin sequence that is at least 60%, 70%, 80%, 90%. 95%, 96%, deacetylase protein family, or preferably to the sir2 family, 97%, 98%, or 99% identical to GenBank Accession Nos. which include yeast Sir2 (GenBank Accession No. P53685), AAHO1042, NP 036371, NP 001017524, or NP 071878, C. elegans Sir-2.1 (GenBank Accession No. NP 501912), and functional fragments thereof. Polypeptides of the inven and human SIRT1 (GenBank Accession No. NM 012238 tion also include homologs (e.g., orthologs and paralogs), and NP 036370 (or AF083106)) and SIRT2 (GenBank variants, or fragments, of GenBank Accession Nos. Accession No. NM 012237, NM 030593, NP 036369, AAHO1042, NP 036371, NP 001017524, or NP 071878. NP 085096, and AF083107) proteins. Other family mem In one embodiment, a SIRT3 protein includes a fragment of bers include the four additional yeast Sir2-like genes termed SIRT3 protein that is produced by cleavage with a mitochon “HST genes' homologues of Sir two) HST1, HST2, HST3 drial matrix processing peptidase (MPP) and/or a mitochon and HST4, and the five other human homologues hSIRT3, drial intermediate peptidase (MIP). US 2009/0143376 A1 Jun. 4, 2009

I0086. The term “substantially homologous' when used in RNA. Also included are vectors that provide more than one of connection with amino acid sequences, refers to sequences the above functions. As used herein, "expression vectors' are which are substantially identical to or similar in sequence defined as polynucleotides which, when introduced into an with each other, giving rise to a homology of conformation appropriate host cell, can be transcribed and translated into a and thus to retention, to a useful degree, of one or more polypeptide(s). An "expression system' usually connotes a biological (including immunological) activities. The term is Suitable host cell comprised of an expression vector that can not intended to imply a common evolution of the sequences. function to yield a desired expression product. 0087. The term “synthetic' is art-recognized and refers to 0094. The term “vision impairment” refers to diminished production by in vitro chemical or enzymatic synthesis. vision, which is often only partially reversible or irreversible 0088. The terms “systemic administration.”“administered upon treatment (e.g., Surgery). Particularly severe vision systemically.” “peripheral administration' and “administered impairment is termed “blindness’ Or “vision loss’, which peripherally are art-recognized and refer to the administra refers to a complete loss of vision, vision worse than 20/200 tion of a Subject composition, therapeutic or other material that cannot be improved with corrective lenses, or a visual other than directly into the central nervous system, Such that field of less than 20 degrees diameter (10 degrees radius). it enters the patient's system and, thus, is Subject to metabo lism and other like processes. 2. Sirtuin Modulators 0089. The term “therapeutic agent' is art-recognized and refers to any chemical moiety that is a biologically, physi 0095. In one aspect, the invention provides novel sirtuin ologically, or pharmacologically active Substance that acts modulating compounds for treating and/or preventing a wide locally or systemically in a Subject. The term also means any variety of diseases and disorders including, for example, dis Substance intended for use in the diagnosis, cure, mitigation, eases or disorders related to aging or stress, diabetes, obesity, treatment or prevention of disease or in the enhancement of neurodegenerative diseases, ocular diseases and disorders, desirable physical or mental development and/or conditions cardiovascular disease, blood clotting disorders, inflamma in an animal or human. tion, cancer, and/or flushing, etc. Sirtuin-modulating com 0090 The term “therapeutic effect” is art-recognized and pounds that increase the level and/or activity of a sirtuin refers to a local or systemic effect in animals, particularly protein may also be used for treating a disease or disorder in mammals, and more particularly humans caused by a phar a subject that would benefit from increased mitochondrial macologically active Substance. The phrase “therapeutically activity, for enhancing muscle performance, for increasing effective amount’ means that amount of such a substance that muscle ATP levels, or for treating or preventing muscle tissue produces some desired local or systemic effect at areasonable damage associated with hypoxia or ischemia. Other com benefit/risk ratio applicable to any treatment. The therapeu pounds disclosed herein may be suitable for use in a pharma tically effective amount of such substance will vary depend ceutical composition and/or one or more methods disclosed ing upon the Subject and disease condition being treated, the herein. weight and age of the Subject, the severity of the disease 0096. In one embodiment, sirtuin-modulating compounds condition, the manner of administration and the like, which of the invention are represented by Structural Formula (I): can readily be determined by one of ordinary skill in the art. For example, certain compositions described herein may be administered in a Sufficient amount to produce a desired (I) effect at a reasonable benefit/risk ratio applicable to such treatment. 0091 “Transcriptional regulatory sequence' is a generic term used throughout the specification to refer to DNA sequences, such as initiation signals, enhancers, and promot ers, which induce or control transcription of protein coding sequences with which they are operable linked. In preferred embodiments, transcription of one of the recombinant genes is under the control of a promoter sequence (or other tran or a salt thereof, where, as Valence permits: Scriptional regulatory sequence) which controls the expres 0097 Ring A is optionally substituted; sion of the recombinant gene in a cell-type which expression 0.098 L is absent, substituted or unsubstituted phenylene, is intended. It will also be understood that the recombinant substituted or unsubstituted —O-phenylene, substituted or gene can be under the control of transcriptional regulatory unsubstituted thienylene, substituted or unsubstituted pyra sequences which are the same or which are different from Zolylene, substituted or unsubstituted benzothiazolylene, those sequences which control transcription of the naturally occurring forms of genes as described herein. NR C(O) NRs , S—, CHR =CHR or 0092 “Treating a condition or disease refers to curing as —CHR C(O)—: well as ameliorating at least one symptom of the condition or 0099 L' is absent, substituted or unsubstituted phenylene, disease. substituted or unsubstituted —O-phenylene, substituted or 0093. A “vector is a self-replicating nucleic acid mol unsubstituted thienylene, substituted or unsubstituted pyra ecule that transfers an inserted nucleic acid molecule into Zolylene, substituted or unsubstituted benzothiazolylene, and/or between host cells. The term includes vectors that substituted or unsubstituted indenedionylene. —C(O)O—, function primarily for insertion of a nucleic acid molecule —C(O)NR —NRC(O)— —NR C(O) NRs , into a cell, replication of vectors that function primarily for —S—, —CHR=CHR, or —CHRs C(O)—, provided the replication of nucleic acid, and expression vectors that that at least one of L and L' is substituted or unsubstituted function for transcription and/or translation of the DNA or phenylene, Substituted or unsubstituted—O-phenylene, Sub US 2009/0143376 A1 Jun. 4, 2009

stituted or unsubstituted thienylene, substituted or unsubsti heterocyclic ring that is optionally substituted with —(C-C, tuted pyrazolylene, substituted or unsubstituted benzothiaz straight or branched alkyl), and wherein at least one of Rao or olylene, substituted or unsubstituted indenedionylene, Rs is not H: 0112 R is selected from —C-C straight or branched —NR , —C(O)O , —C(O)NR , NRC(O)—, alkylene or —C(O)—, and NR C(O) NRs , S—, CHR =CHR, or 0113 each of ring K and ring E is independently substi —CHRs C(O)—: tuted with up to three substituents independently selected 0100 R is absent, —H, NRRs—NC(O)Rs, ORs, from halo. —CF, —O—(C-C straight or branched alkyl). naphthyl or a heterocyclic group, provided that L and R are —S—(C-C straight or branched alkyl), —N(Rao)(Rs), not both absent unless X is N: —S(O), N(Ra)(Rs), heterocyclyl (C-C straight or branched alkyl)-heterocyclyl. —O—(C-C straight or 0101 R is —H, unsubstituted alkyl, - NRRs, NRC branched alkyl)-heterocyclyl. —S-(C-C straight or (O)Rs —ORs, substituted or unsubstituted phenyl, naphthyl branched alkyl)-heterocyclyl, or is optionally fused to a 5-6 or a heterocyclic group; membered heterocyclyl or heteroaryl, wherein any heterocy 0102 R is —H, NRRs, NC(O)Rs —ORs or a sub clyl or heteroaryl is optionally substituted with —C-C, stituted or unsubstituted heterocyclic group, or R and Rs. straight or branched alkyl. taken together with the atoms to which they are attached, form 0114. In certain embodiments, one of Rao or Rso is H. an optionally Substituted heterocyclic group, or R is absent 0.115. In certain embodiments, ring K is substituted with when Z is O or S: up to 3 substituents independently selected from methyl, 0103 Ra and Rs are independently —H, a substituted or —O-methyl, - N(CH), or —CF, but is unsubstituted in unsubstituted alkyl group, a Substituted or unsubstituted aryl the positions ortho to the attachment to the rest of the mol group or a Substituted or unsubstituted non-aromatic hetero ecule. cyclic group: 0116. In certain embodiments, such as where Rao and/or Rs have the values indicated above and/or ring K has the 0104 R. R., and Rs are independently selected from the substitution pattern described above, ring E is substituted group consisting of halogen, —OR, —CN, —COR, with up to 2 substituents independently selected from methyl, —OCOR —OCOR —C(O)NRRs —OC(O)NRRs, —O-methyl, -S(O), N(CH), —O-methyl-morpholino, —C(O)R. —COR —SR —OSOH, -S(O), R —S(O) —O-ethyl-morpholino, fluoro. —CF, piperidyl, methylpip OR —S(O)NRRs —NRRs —NRC(O)ORs, —NRC eridyl, pyrrolidyl, or methylpyrrolidyl. (O)Rs and—NO; I0117. In certain embodiments, Ro is selected from —H, 0105 W is C or N: —CH2-piperazinyl. —CH2-methylpiperazinyl, -CH2-pyr rolidyl, —CH-piperidyl, -CH2-morpholino. —CH2—N 0106 X is C or N: (CH), —C(O)—NH-(CH2)-piperazinyl. —C(O)— 01.07 Y is C or N: NH-(CH)-methylpiperazinyl, —C(O) NH-(CH)- 0108 Z is C, N, O or S, provided that at least two of W, X, pyrrolidyl, —C(O) NH (CH)-morpholino. —C(O)— Y and Z are C; and NH-(CH)-piperidyl, or —C(O) NH-(CH)N(CH), 0109 n is 1 or 2. wherein n is 1 or 2. 0118. In particular embodiments, ring K is substituted 0110. In a second embodiment, sirtuin-modulating com with up to 3 substituents independently selected from methyl, pounds of the invention are represented by Structural For O-methyl, N(CH), CF, but is unsubstituted in the positions mula (Ia): ortho to the attachment to the rest of the molecule; ring E is substituted with up to 2 substituents independently selected from methyl, O-methyl, S(O), N(CH), —O-methyl (Ia) morpholino. —O-ethyl-morpholino, fluoro. —CF, meth ylpiperidyl, or pyrrolidyl; and Rio is selected from —H. —CH-piperazinyl. —C(O)—NH-(CH2)-piperazinyl. —C(O) NH (CH-)-methylpiperazinyl, —C(O)—NH (CH)-pyrrolidyl, or —C(O) NH-(CH), N(CH). R11 0119. In a further embodiment, sirtuin-modulating com V NH pounds of the invention are represented by Formula (Ib):

21 NN N (Ib) N1's N s

R11 V wherein: NH 0111 Ro is selected from —H. —C(O)—N(R)(Rs), —S(O)N(Rao)(Rs), or —CH2—N(R)(Rs); wherein each of Rao and Rso is independently selected from —H. —C-C straight or branched alkyl, —(C-C straight or 1) (r. branched alkyl)-N(CH), —(C-C straight or branched 13 alkyl)-heterocyclyl. —(C-C straight or branched alkyl)- alkylheterocyclyl, or wherein Rao and Rso taken together with the Natom to which they are bound form a 5-6 membered US 2009/0143376 A1 Jun. 4, 2009

wherein: 0.134 One group of compounds of the invention encom 0120 Z is selected from O or S: passed by Structural Formula (II) is represented by Structural 0121 Rio is selected from —H. —C(O)—N(R)(Rs), Formula (III): —S(O)N(Rao)(Rs), or —CH2—N(Rao)(Rs); wherein each of Rao and Rs is independently selected from —H. (III) —C-C straight or branched alkyl, —(C-C straight or O branched alkyl)-N(CH), —(C-C straight or branched alkyl)-heterocyclyl. —(C-C straight or branched alkyl)- alkylheterocyclyl, or wherein Rao and Rso taken together with *s-seN Y N1,N the Natom to which they are bound form a 5-6 membered D s heterocyclic ring that is optionally substituted with —(C-C, O) () straight or branched alkyl), and wherein at least one of Rao or Rso is not H. or a salt thereof, where: 0122 R is selected from —C-C straight or branched 0.135 Rings D and E are optionally substituted; and alkylene or —C(O)—: 0.136 R. is —H, a substituted or unsubstituted alkyl 0123 each of R and R is independently selected from group, a Substituted or unsubstituted aryl group or a Substi —H or —(C-C straight or branched alkyl), or R and R. tuted or unsubstituted non-aromatic heterocyclic group. are taken together to form a benzene ring that is Substituted 0.137 In certain embodiments, Ring E is substituted with with up to two substituents independently selected from an acylamino, heterocyclylcarbonylamino, lower alkyl or —(C-C straight or branched allyl), —CF or halo; and Substituted or unsubstituted alkoxy group. 0.124 ring K is substituted with up to three substituents 0.138. In certain embodiments, Ring D is substituted with independently selected from halo. —CF, —O—(C-C, an amino group. In particular embodiments, Ring E is Sub straight or branched alkyl). —S (C-C straight or branched stituted with an acylamino, heterocyclylcarbonylamino, alkyl), —N(Rao)(Rs). —S(O) N(Rao)(Rs), heterocyclyl, lower alkyl, or Substituted or unsubstituted alkoxy group, and (C-C straight or branched allyl)-heterocyclyl. —O—(C- Ring D is Substituted with an amino group. C. Straight or branched alkyl)-heterocyclyl. —S (C-C, 0.139. In certain embodiments, R is a substituted alkyl straight or branched alkyl)-heterocyclyl, or is optionally group. fused to a 5-6 membered heterocyclyl or heteroaryl, wherein 0140. A group of compounds of the invention encom any heterocyclyl or heteroaryl is optionally substituted with passed by Structural Formula (III) is represented by Struc —C-C straight or branched alkyl. tural Formula (IV): 0.125. In certain embodiments, Ro is —H. 0126. In certain embodiments, ring K is substituted with up to 3 substituents independently selected from methyl, (IV) O-methyl, N(CH), CF, and whereinring K is unsubstituted in the positions ortho to the attachment to the rest of the molecule. ---, 0127. In certain embodiments, such as when Rio is —H Cy-K) and/or ring K has the substitution pattern described above, C. each of R and R is independently selected from —H. methyl, -O-methyl, —S(O), N(CH), —O-methyl-mor pholino, —O-ethyl-morpholino, fluoro. —CF, piperidyl, or a salt thereof, where: methylpiperidyl, pyrrolidyl, or methylpyrrolidyl. 0141 Ring E is optionally substituted; and 0128. In a preferred embodiment, each of R and R is I0142 R and Rs are independently —H, a substituted or methyl. unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic hetero 0129. In another embodiment, sirtuin-modulating com cyclic group. pounds of the invention are represented by Formula (II): 0143. In certain embodiments, Ring E is substituted with an acylamino, heterocyclylcarbonylamino, lower alkyl or (II) Substituted or unsubstituted alkoxy group. 0144. In certain embodiments, R is a substituted alkyl group. In particular embodiments, Ring E is substituted with an acylamino, heterocyclylcarbonylamino, lower alkyl or Substituted or unsubstituted alkoxy group and R is a substi tuted alkyl group. (0145. In certain embodiments, R is a substituted or unsubstituted alkyl group. Such as an aralkyl or a cycloalkyl or a salt thereof, where: group (e.g., benzyl, cyclohexyl). In particular embodiments, 0130 Rings C, D and E are optionally substituted; and Rs is a substituted or unsubstituted alkyl group when Ring E 0131 x is 0 or 1. is substituted with an acylamino, heterocyclylcarbony 0132. In certain embodiments X is 0. lamino, lower alkyl or substituted or unsubstituted alkoxy 0133. In certain embodiments (e.g., where x is 0), Ring C group and/or R is a Substituted alkyl group. is Substituted with a group that is capable of providing a trans 0146. One group of compounds of the invention encom configuration (e.g., an amide group, an optionally 2-substi passed by Structural Formula (IV) is represented by Struc tuted 1-alkenyl group). tural Formula (IVa): US 2009/0143376 A1 Jun. 4, 2009 10

0154) A group of compounds of the invention encom passed by Structural Formula (V) are represented by Struc (IVa) Rs tural Formula (VI): V NH (VI) Rs 21 NN N V NH O N1SN s 21 N N NHX- R9, O NH V N1SN R4 R4 n N O or a salt thereof. H 0147 Another group of compounds of the invention encompassed by Structural Formula (IV) is represented by or a salt thereof, where: Structural Formula (IVb): 0155 R. Rs and R are independently —H, a substituted or unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic (IVb) Rs heterocyclic group. O V I0156. In certain embodiments, R is a substituted alkyl NH group. R4 n. 21 0157. In certain embodiments, Rs is a substituted or N N1 N s unsubstituted alkyl group. Such as an aralkyl or a cycloalkyl group (e.g., benzyl, cyclohexyl). In particular embodiments, N s Rs is a Substituted or unsubstituted alkyl group and R is a substituted alkyl group. or a salt thereof. 0158. In certain embodiments, R is a C alkyl group 0148 Yet another group of compounds of the invention (e.g., methyl, cyclopropyl), a substituted or unsubstituted aryl encompassed by Structural Formula (IV) is represented by group (e.g., Substituted or unsubstituted phenyl) or a Substi Structural Formula (V): tuted or unsubstituted non-aromatic heterocyclic group (e.g., furanyl, morpholino). In particular embodiments, R is a Ca alkyl group, a Substituted or unsubstituted aryl group or a (V) Substituted or unsubstituted non-aromatic heterocyclic group O RSN when Rs is a Substituted or unsubstituted allyl group and/or R is a Substituted alkyl group 's--a 0159. In yet another embodiment, sirtuin-modulating Cy-Kyr compounds of the invention are represented by Formula C. \ / (VII):

0149 or a salt thereof, where: (VII) 0150. R. R. and Rs are independently —H, a substituted R3 or unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic heterocyclic group. L 0151. In certain embodiments, R is a substituted alkyl C3N group. 0152. In certain embodiments, Rs is a substituted or or a salt thereof, where: unsubstituted alkyl group. Such as an aralkyl or a cycloalkyl group (e.g., benzyl, cyclohexyl). In particular embodiments, (0160 Ring F is optionally substituted; Rs is a Substituted or unsubstituted alkyl group and R is a 0.161 L' is substituted or unsubstituted phenylene, substi Substituted alkyl group. tuted or unsubstituted thienylene, substituted or unsubstituted 0153. In certain embodiments, R is a C alkyl group indonedionylene. —C(O)O— —NRC(O)— —S—, (e.g., methyl, cyclopropyl), a substituted or unsubstituted aryl —CHR=CHR, or —CHRs C(O) ; group (e.g., Substituted or unsubstituted phenyl) or a Substi 0162 R is —H, unsubstituted alkyl, - NRRs —NRC tuted or unsubstituted non-aromatic heterocyclic group (e.g., (O)Rs —ORs, substituted or unsubstituted phenyl or a het furanyl, morpholino). In particular embodiments, R is a Ca erocyclic group; allyl group, a Substituted or unsubstituted aryl group or a 0163 R is —H. —NRRs —NC(O)Rs —ORs or a het Substituted or unsubstituted non-aromatic heterocyclic group erocyclic group, or R and R, taken together with the atoms when Rs is a Substituted or unsubstituted alkyl group and/or to which they are attached, form an optionally substituted R is a Substituted alkyl group heterocyclic group, or R is absent when Z is O or S; US 2009/0143376 A1 Jun. 4, 2009

0164 Ra and Rs are independently —H, a substituted or 0179. In particular embodiments, L' is substituted or unsubstituted alkyl group, a Substituted or unsubstituted aryl unsubstituted —O-phenylene or —CHRs C(O)—. In such group or a Substituted or unsubstituted non-aromatic hetero embodiments, R is preferably a heterocyclic group. cyclic group: 0180. In particular embodiments, L' is a substituted or 0.165 R. R., and Rs are independently selected from the unsubstituted thienylene. In Such embodiments, R is group consisting of halogen, —OR, —CN, —COR, - NRC(O)Rs. —OCOR —OCOR —C(O)NRRs —OC(O)NRRs, 0181 Another group of compounds of the invention —C(O)R. —COR —SR —OSOH, -S(O), R —S(O) encompassed by Structural Formula (VII) is represented by OR - S(O)NRRs, NRRs, NRC(O)ORs, NRC Structural Formula (IX): (O)Rs and—NO; (0166 Z is C, N, O or S; and (IX) (0167 n is 1 or 2. R 0.168. In Structural Formula (VII), the dashed bond indi cated as ------represents a single or double bond. For the \ 3 R2, compounds represented by Structural Formula (VII), both X-1/ dashed bonds cannot be double bonds, but preferably one of N the dashed bonds is a double bond and the other is a single bond. 0169. One group of compounds encompassed by Struc or a salt thereof, where: tural Formula (VII) are represented by Structural Formula 0182 Ring H is optionally substituted; (VIII): 0183 L' is substituted or unsubstituted phenylene. —S or —CHR =CHR, ; 0184 R is NRRs, NRC(O)Rs or a heterocyclic (VIII) group; S R2, 0185. R is —H, or R and R, taken together with the / atoms to which they are attached, form an optionally Substi A tuted heterocyclic group; N 0186 R and Rs are independently —H, a substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl or a salt thereof, where: group or a Substituted or unsubstituted non-aromatic hetero 0170 Ring G is optionally substituted; cyclic group; 0171 L' is substituted or unsubstituted phenylene, substi 0187 RandR, are independently selected from the group tuted or unsubstituted —O-phenylene, substituted or unsub consisting of halogen, —OR, —CN, —COR —OCOR stituted thienylene, substituted or unsubstituted indonedio —OCOR —C(O)NRRs —OC(O)NRRs, —C(O)R. nylene, —NRC(O) , C(O)O , S , COR, SR —OSOH, -S(O), R —S(O), OR, —CHR=CHR, or —CHRs C(O) ; —S(O)NRRs —NRRs, NRC(O)ORs, NRC(O)Rs 0172 R is —H, unsubstituted alkyl, - NRRs —NRC and —NO, and (O)Rs —ORs, substituted or unsubstituted phenyl or a het 0188 n is 1 or 2. erocyclic group; 0189 In certain embodiments, L' is —S . In particular (0173 R and Rs are independently —H, a substituted or embodiments, L' is —S- and R and R, taken together with unsubstituted alkyl group, a Substituted or unsubstituted aryl the atoms to which they are attached, form an optionally group or a Substituted or unsubstituted non-aromatic hetero Substituted heterocyclic group. cyclic group: 0190. In certain embodiments, L' is a substituted or unsub 0.174 R. R., and Rs are independently selected from the stituted phenylene. In particular embodiments, L' is a Substi group consisting of halogen, —OR, —CN. —COR tuted or unsubstituted phenylene and R is NRC(O)Rs. —OCOR —OCOR —C(O)NRRs —OC(O)NRRs, 0191 In certain embodiments, L' is —CHR =CHR, , —C(O)R. —COR - SR —OSOH, -S(O), R = S(O) such as —CH2—CH2—, or—C(CN)—CH2—. In particular OR —S(O)NRRs —NRRs —NRC(O)ORs, —NRC embodiments, L' is —CHR=CHR, and R is NRR or (O)Rs and —NO; and a Substituted or unsubstituted aryl group. (0175 n is 1 or 2. 0.192 A further group of compounds of the invention 0176). In certain embodiments, L' is substituted or unsub encompassed by Structural Formula (VII) is represented by stituted —O-phenylene, substituted or unsubstituted thie Structural Formula (X): nylene or —CHRs C(O)—. 0177. In certain embodiments, R is NRC(O)Rs or a heterocyclic group, such as NRC(O)-substituted alkyl. In R (X) a particular embodiment, R is NRC(O)Rs or a heterocy - “. clic group and L' is substituted or unsubstituted —O-phe \ R22 nylene, substituted or unsubstituted thienylene or —CHRs – / C(O)-. / L 0178. In certain embodiments, Ring G is unsubstituted. In a particular embodiment, Ring G is unsubstituted when R is —NRC(O)Rs or a heterocyclic group and/or L is substituted R or unsubstituted—O-phenylene, substituted or unsubstituted thienylene or —CHRs C(O)—. US 2009/0143376 A1 Jun. 4, 2009

or a salt thereof, where: (e.g., glucuronate, glucose) derivatives. In other words, Sub 0193 Ring J is optionally substituted; stituent groups —OH also include —OSOM", where M is 0194 L is substituted or unsubstituted phenylene, substi a suitable cation (preferably H, NH, or an alkali metal ion tuted or unsubstituted —O-phenylene, substituted or unsub such as Na' or K) and Sugars such as stituted thienylene, substituted or unsubstituted pyrazolylene,

substituted or unsubstituted benzothiazolylene. —C(O)O , —C(O)NR —NRC(O)— —NR C(O) NRs , HOC —S , —CHR =CHR, or —CHR C(O)—; 0.195 L is substituted or unsubstituted phenylene, substi tuted or unsubstituted —O-phenylene, substituted or unsub HO and stituted thienylene, substituted or unsubstituted pyrazolylene, substituted or unsubstituted benzothiazolylene. —C(O)O , —C(O)NR , NRC(O)— —NR C(O) NRs , —S , —CHR =CHR, or —CHRs C(O)—; 0.196 R is —H. —NRRs —NC(O)Rs —ORs, naph thyl or a heterocyclic group; OH 0.197 R is —H, unsubstituted alkyl, - NRRs —NRC (O)Rs —ORs, naphthyl or a heterocyclic group: 0198 R is —H. —NRRs, NC(O)Rs —ORs or a sub stituted or unsubstituted heterocyclic group; 0199 Ra and Rs are independently —H, a substituted or Ho unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic hetero cyclic group: These groups are generally cleavable to —OH by hydrolysis 0200 R. R., and Rs are independently selected from the or by metabolic (e.g., enzymatic) cleavage. group consisting of halogen, —OR, —CN, —COR, 0211. In certain embodiments, the compounds of the —OCOR —OCOR —C(O)NRRs —OC(O)NRRs, invention exclude one or more of Compounds 1-90. —C(O)R. —COR - SR —OSOH, -S(O), R = S(O) 0212 Sirtuin-modulating compounds of the invention OR - S(O)NRRs, NRRs, NRC(O)ORs, NRC advantageously modulate the level and/or activity of a sirtuin (O)Rs and - NO; and protein, particularly the deacetylase activity of the sirtuin 0201 n is 1 or 2. protein. 0202 In certain embodiments, L is NRC(O)—. 0213 Separately or in addition to the above properties, 0203. In certain embodiments, R is a substituted or certain sirtuin-modulating compounds of the invention do not unsubstituted aryl group. In particular embodiments, R is a substantially have one or more of the following activities: substituted or unsubstituted aryl group and L is —NRC inhibition of PI3-kinase, inhibition of aldoreductase, inhibi (O)—. tion of tyrosine kinase, transactivation of EGFR tyrosine 0204. In certain embodiments, L' is —C(O)O—. In par kinase, coronary dilation, or spasmolytic activity, at concen trations of the compound that are effective for modulating the ticular embodiments, L' is —C(O)O— when R is a substi deacetylation activity of a sirtuin protein (e.g., Such as a tuted or unsubstituted aryl group and/or L is —NRC(O)—. SIRT1 and/or a SIRT3 protein). 0205. In certain embodiments, R is an unsubstituted alkyl 0214. An alkyl group is a straight chained, branched or group. In particular embodiments, R is an unsubstituted cyclic non-aromatic hydrocarbon which is completely satu alkyl group when L' is —C(O)O R is a substituted or rated. Typically, a straight chained or branched alkyl group unsubstituted aryl group and/or L is —NRC(O)—. has from 1 to about 20 carbon atoms, preferably from 1 to 0206. In certain embodiments, Ring G is substituted, such about 10, and a cyclic alkyl group has from 3 to about 10 as with one or more (e.g., two) alkoxy (e.g., methoxy, ethoxy) carbon atoms, preferably from 3 to about 8. Examples of groups, for example, in the positions ortho to the bridgehead straight chained and branched alkyl groups include methyl, carbon atoms. In particular embodiments, Ring G is Substi ethyl, n-propyl, iso-propyl. n-butyl, sec-butyl, tert-butyl, pen tuted when R is an unsubstituted alkyl group, L' is —C(O) tyl, hexyl, pentyl and octyl. A C-C straight chained or O—, R is a Substituted or unsubstituted aryl group and/or L branched alkyl group is also referred to as a “lower alkyl is NRC(O)—. group. 0207 Novel compounds of the invention can also be used 0215. An alkenyl group is a straight chained, branched or in the methods described herein. cyclic non-aromatic hydrocarbon which contains one or more 0208. The compounds and salts thereof described herein double bonds. Typically, the double bonds are not located at also include their corresponding hydrates (e.g., hemihydrate, the terminus of the alkenyl group, such that the double bond monohydrate, dihydrate, trihydrate, tetrahydrate) and sol is not adjacent to another functional group. 0216. An alkynyl group is a straight chained, branched or vates. Suitable solvents for preparation of solvates and cyclic non-aromatic hydrocarbon which contains one or more hydrates can generally be selected by a skilled artisan. triple bonds. Typically, the triple bonds are not located at the 0209. The compounds and salts thereof can be present in terminus of the alkynyl group, such that the triple bond is not amorphous or crystalline (including co-crystalline and poly adjacent to another functional group. morph) forms. 0217. A cyclic group includes carbocyclic and heterocy 0210 Sirtuin-modulating compounds of the invention clic rings. Such rings can be saturated or unsaturated, includ having hydroxyl Substituents, unless otherwise indicated, ing aromatic. Heterocyclic rings typically contain 1 to 4 het also include the related secondary metabolites, particularly eroatoms, although oxygen and Sulfur atoms cannot be Sulfate, phosphate, acyl (e.g., acetyl, fatty acid acyl) and Sugar adjacent to each other. US 2009/0143376 A1 Jun. 4, 2009

0218. Aromatic (aryl) groups include carbocyclic aro 0223) A hydrogen-bond donating group is a functional matic groups such as phenyl, naphthyl, and anthracyl, and group having a partially positively-charged hydrogen atom heteroaryl groups such as imidazolyl, thienyl, furanyl. (e.g., -OH, -NH2. —SH) or a group (e.g., an ester) that pyridyl, pyrimidyl, pyranyl, pyrazolyl pyrroyl, pyrazinyl, metabolizes into a group capable of donating a hydrogen thiazolyl, oxazolyl, and tetrazolyl. bond. 0219. Aromatic groups also include fused polycyclic aro 0224 Combinations of substituents and variables envi matic ring systems in which a carbocyclic aromatic ring or sioned by this invention are only those that result in the heteroaryl ring is fused to one or more other heteroaryl rings. formation of stable compounds. As used herein, the term Examples include benzothienyl, benzofuranyl, indolyl, “stable' refers to compounds that possess stability sufficient to allow manufacture and that maintain the integrity of the quinolinyl, benzothiazole, benzooxazole, benzimidazole, compound for a sufficient period of time to be useful for the quinolinyl, isoquinolinyl and isoindolyl. purposes detailed herein. 0220 Non-aromatic heterocyclic rings are non-aromatic 0225. Double bonds indicated in a structure as: carbocyclic rings which include one or more heteroatoms Such as nitrogen, oxygen or Sulfur in the ring. The ring can be five, six, seven or eight-membered. Examples include tet rry rahydrofuranyl, tetrahydrothiophenyl, morpholino, thiomor pholino, pyrrolidinyl, piperazinyl, piperidinyl, and thiazolidi nyl, along with the cyclic form of Sugars. are intended to include both the (E)- and (Z)-configuration. 0221. A ring fused to a second ring shares at least one Preferably, double bonds are in the (E)-configuration. common bond. 0226. A sugar is an aldehyde or ketone derivative of a 0222 Suitable substituents on an alkyl, alkenyl, alkynyl, straight-chain polyhydroxy alcohol, which contains at least aryl, non-aromatic heterocyclic or aryl group (carbocyclic three carbon atoms. A Sugar can exist as a linear molecule or, and heteroaryl) are those which do not substantially interfere preferably, as a cyclic molecule (e.g., in the pyranose or with the ability of the disclosed compounds to have one or furanose form). Preferably, a Sugar is a monosaccharide Such more of the properties disclosed herein. A substituent sub as glucose or glucuronic acid. In embodiments of the inven stantially interferes with the properties of a compound when tion where, for example, prolonged residence of a compound the magnitude of the property is reduced by more than about derivatized with a Sugar is desired, the Sugar is preferably a 50% in a compound with the substituent compared with a non-naturally occurring Sugar. For example, one or more compound without the substituent. Examples of suitable sub hydroxyl groups are substituted with another group, such as a stituents include —OH, halogen ( Br, —Cl. —I and —F), halogen (e.g., chlorine). The stereochemical configuration at OR', O COR, COR, C(O)R, CN, NO, one or more carbonatoms can also be altered, as compared to -COOH,-COOR, OCOR, C(O)NR'R'', OC(O) a naturally occurring Sugar. One example of a Suitable non NR'R', SOH, NH, NHR, N(RR), COOR, naturally occurring Sugar is Sucralose. CHO, CONH, CONHR, CONCRR), NH 0227. A fatty acid is a carboxylic acid having a long COR, NRCOR, NHCONH, NHCONRH, NH chained hydrocarbon moiety. Typically, a fatty acid has an CONGRR), NRCONH, NRCONRH, NRCON even number of carbon atoms ranging from 12 to 24, often (RR), —C(=NH) NH, C(-NH) NHR, from 14 to 20. Fatty acids can be saturated or unsaturated and C(-NH) N(RR), -C(=NR) NH, substituted or unsubstituted, but are typically unsubstituted. C(-NR). NHR, C(-NR) N(RR), NH C Fatty acids can be naturally or non-naturally occurring. In (—NH) NH, NH C(-NH) NHR, NH C embodiments of the invention where, for example, prolonged (—NH) N(RR), NH C(=NR) NH, -NH C residence time of a compound having a fatty acid moiety is (—NR). NHR, NH C(-NR) N(RR), desired, the fatty acid is preferably non-naturally occurring. NRH C(-NH) NH, NR C(-NH) NHR, The acyl group of a fatty acid consists of the hydrocarbon NR C(-NH) N(RR), NR C(-NR) NH, moiety and the carbonyl moiety of the carboxylic acid func NR C(-NR) NHR', NR C(—NR) N tionality, but excludes the –OH moiety associated with the (RR), NHNH, NHNHR, NHR'R', SONH, carboxylic acid functionality. —SONHR, SONR'R'', CH=CHR, 0228. Also included in the present invention are salts, CH=CRR, CR CRR, CR -CHR, particularly pharmaceutically acceptable salts, of the sirtuin CR-CRR, CCR, SH, SOR" (k is 0, 1 or 2), modulating compounds described herein. The compounds of —S(O), OR (k is 0, 1 or 2) and NH C(=NH) NH. the present invention that possess a Sufficiently acidic, a Suf R-R are each independently an aliphatic, substituted ali ficiently basic, or both functional groups, can react with any phatic, benzyl, substituted benzyl, aromatic or substituted of a number of inorganic bases, and inorganic and organic aromatic group, preferably an alkyl, benzylic oraryl group. In acids, to form a salt. Alternatively, compounds that are inher addition, NR'R'', taken together, can also form a substi ently charged. Such as those with a quaternary nitrogen, can tuted or unsubstituted non-aromatic heterocyclic group. A form a salt with an appropriate counterion (e.g., a halide Such non-aromatic heterocyclic group, benzylic group or aryl as bromide, chloride, or fluoride, particularly bromide). group can also have analiphatic or Substituted aliphatic group 0229. Acids commonly employed to form acid addition as a substituent. A substituted aliphatic group can also have a salts are inorganic acids such as hydrochloric acid, hydrobro non-aromatic heterocyclic ring, a Substituted a non-aromatic mic acid, hydroiodic acid, Sulfuric acid, phosphoric acid, and heterocyclic ring, benzyl, Substituted benzyl, aryl or Substi the like, and organic acids Such as p-toluenesulfonic acid, tuted aryl group as a Substituent. A substituted aliphatic, methanesulfonic acid, oxalic acid, p-bromophenyl-Sulfonic non-aromatic heterocyclic group, Substituted aryl, or Substi acid, carbonic acid, Succinic acid, citric acid, benzoic acid, tuted benzyl group can have more than one substituent. acetic acid, and the like. Examples of Such salts include the US 2009/0143376 A1 Jun. 4, 2009

sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, have an ECso for activating a human sirtuin, Such as SIRT1 monohydrogenphosphate, dihydrogenphosphate, metaphos and/or SIRT3, deacetylase activity that is at least 5 fold less phate, pyrophosphate, chloride, bromide, iodide, acetate, pro than the ECso for activating a yeast sirtuin, such as Sir2 (Such pionate, decanoate, caprylate, acrylate, formate, isobutyrate, as Candida, S. cerevisiae, etc.), and even more preferably at caproate, heptanoate, propiolate, oxalate, malonate. Succi least 10 fold, 100 fold or even 1000 fold less. In another nate, Suberate, sebacate, fumarate, maleate, butyne-1,4-dio embodiment, an inhibitor of a sirtuin protein from lower ate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methyl eukaryotes, particularly yeast or human pathogens, does not benzoate, dinitrobenzoate, hydroxybenzoate, have any substantial ability to inhibit a sirtuin protein from methoxybenzoate, phthalate, Sulfonate, Xylenesulfonate, humans at concentrations (e.g., in vivo) effective for inhibit phenylacetate, phenylpropionate, phenylbutyrate, citrate, ing the deacetylase activity of a sirtuin protein from a lower lactate, gamma-hydroxybutyrate, glycolate, tartrate, meth eukaryote. For example, a sirtuin-inhibiting compound may anesulfonate, propanesulfonate, naphthalene-1-sulfonate, be chosen to have an ICso for inhibiting a human sirtuin, Such naphthalene-2-sulfonate, mandelate, and the like. as SIRT1 and/or SIRT3, deacetylase activity that is at least 5 0230 Base addition salts include those derived from inor fold less than the ICso for inhibiting a yeast sirtuin, Such as ganic bases, such as ammonium or alkali or alkaline earth Sir2 (such as Candida, S. cerevisiae, etc.), and even more metal hydroxides, carbonates, bicarbonates, and the like. preferably at least 10 fold, 100 fold or even 1000 fold less. Such bases useful in preparing the salts of this invention thus 0235. In certain embodiments, a sirtuin-modulating com include Sodium hydroxide, potassium hydroxide, ammonium pound may have the ability to modulate one or more sirtuin hydroxide, potassium carbonate, and the like. protein homologs, such as, for example, one or more or 0231. In an exemplary embodiment, a sirtuin-modulating human SIRT1, SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, or compound may traverse the cytoplasmic membrane of a cell. SIRT7. In one embodiment, a sirtuin-modulating compound For example, a compound may have a cell-permeability of at has the ability to modulate both a SIRT1 and a SIRT3 protein. least about 20%, 50%, 75%, 80%, 90% or 95%. 0236. In other embodiments, a SIRT1 modulator does not 0232 Sirtuin-modulating compounds described herein have any substantial ability to modulate other sirtuin protein may also have one or more of the following characteristics: homologs, such as, for example, one or more of human the compound may be essentially non-toxic to a cell or Sub SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, or SIRT7, at concen ject; the sirtuin-modulating compound may be an organic trations (e.g., in vivo) effective for modulating the deacety molecule or a small molecule of 2000 amu or less, 1000 amu lase activity of human SIRT1, For example, a sirtuin-modu or less; a compound may have a half-life under normal atmo lating compound may be chosen to have an EDso for spheric conditions of at least about 30 days, 60 days, 120 modulating human SIRT1 deacetylase activity that is at least days, 6 months or 1 year, the compound may have a half-life 5 fold less than the EDs for modulating one or more of in solution of at least about 30 days, 60 days, 120 days, 6 human SIRT2, SIRT3, SIRT4, SIRT5, SIRT6, or SIRT7, and months or 1 year; a sirtuin-modulating compound may be even more preferably at least 10 fold, 100 fold or even 1000 more stable in solution than resveratrol by at least a factor of fold less. In one embodiment, a SIRT1 modulator does not about 50%, 2 fold, 5 fold, 10 fold, 30 fold, 50 fold or 100 fold; have any substantial ability to modulate a SIRT3 protein. a sirtuin-modulating compound may promote deacetylation 0237. In other embodiments, a SIRT3 modulator does not of the DNA repair factor Ku70; a sirtuin-modulating com have any substantial ability to modulate other sirtuin protein pound may promote deacetylation of RelA/p65; a compound homologs, such as, for example, one or more of human may increase general turnover rates and enhance the sensitiv SIRT1, SIRT2, SIRT4, SIRT5, SIRT6, or SIRT7, at concen ity of cells to TNF-induced apoptosis. trations (e.g., in vivo) effective for modulating the deacety 0233. In certain embodiments, a sirtuin-modulating com lase activity of human SIRT3. For example, a sirtuin-modu pound does not have any substantial ability to inhibit a histone lating compound may be chosen to have an EDso for deacetylase (HDACs) class I, a HDAC class II, or HDACs I modulating human SIRT3 deacetylase activity that is at least and II, at concentrations (e.g., in vivo) effective for modulat 5 fold less than the EDso for modulating one or more of ing the deacetylase activity of the sirtuin. For instance, in human SIRT1, SIRT2, SIRT4, SIRT5, SIRT6, or SIRT7, and preferred embodiments the sirtuin-modulating compound is a even more preferably at least 10 fold, 100 fold or even 1000 sirtuin-activating compound and is chosen to have an ECso for fold less. In one embodiment, a SIRT3 modulator does not activating sirtuin deacetylase activity that is at least 5 fold less have any substantial ability to modulate a SIRT1 protein. than the ECs for inhibition of an HDAC I and/or HDAC II, 0238. In certain embodiments, a sirtuin-modulating com and even more preferably at least 10 fold, 100 fold or even pound may have a binding affinity for a sirtuin protein of 1000 fold less. Methods for assaying HDAC I and/or HDAC about 10M, 10M, 10M, 10'’M or less. A sirtuin II activity are well known in the art and kits to perform such modulating compound may reduce (activator) or increase assays may be purchased commercially. See e.g., BioVision, (inhibitor) the apparent Km of a sirtuin protein for its sub Inc. (Mountain View, Calif.; world wide web at biovision. strate or NAD+ (or other cofactor) by a factor of at least about com) and Thomas Scientific (Swedesboro, N.J.; world wide 2, 3, 4, 5, 10, 20, 30, 50 or 100. In certain embodiments, Km web at tomas Sci.com). values are determined using the mass spectrometry assay 0234. In certain embodiments, a sirtuin-modulating com described herein. Preferred activator compounds reduce the pound does not have any Substantial ability to modulate sir Km of a sirtuin for its substrate or cofactor to a greater extent tuin homologs. In one embodiment, an activator of a human than caused by resveratrol at a similar concentration or reduce sirtuin protein may not have any Substantial ability to activate the Km of a sirtuin for its substrate or cofactor similar to that a sirtuin protein from lower eukaryotes, particularly yeast or caused by resveratrol at a lower concentration. A sirtuin human pathogens, at concentrations (e.g., in vivo) effective modulating compound may increase the Vmax of a sirtuin for activating the deacetylase activity of human sirtuin. For protein by a factor of at least about 2, 3, 4, 5, 10, 20, 30, 50 or example, a sirtuin-activating compound may be chosen to 100. A sirtuin-modulating compound may have an ED50 for US 2009/0143376 A1 Jun. 4, 2009 modulating the deacetylase activity of a SIRT1 and/or SIRT3 and/or activity of a sirtuin protein may be administered with protein of less than about 1 nM. less than about 10 nM, less one or more of the following compounds: resveratrol, butein, than about 100 nM, less than about 1 uM, less than about 10 fisetin, piceatannol, or quercetin. In an exemplary embodi uM, less than about 100 uM, or from about 1-10 nM, from ment, a sirtuin-modulating compound that increases the level about 10-100 nM, from about 0.1-1 uM, from about 1-10 uM and/or activity of a sirtuin protein may be administered in or from about 10-100 uM. A sirtuin-modulating compound combination with nicotinic acid. In another embodiment, a may modulate the deacetylase activity of a SIRT1 and/or sirtuin-modulating compound that decreases the level and/or SIRT3 protein by a factor of at least about 5, 10, 20, 30, 50, or activity of a sirtuin protein may be administered with one or 100, as measured in a cellular assay or in a cell based assay. A more of the following compounds: nicotinamide (NAM), sirtuin-activating compound may cause at least about 10%, suranim; NF023 (a G-protein antagonist); NF279 (a puriner 30%, 50%, 80%, 2 fold, 5 fold, 10 fold, 50 fold or 100 fold gic receptor antagonist); Trolox (6-hydroxy-2.5.7.8, tetram greater induction of the deacetylase activity of a sirtuin pro ethylchroman-2-carboxylic acid); (-)-epigallocatechin (hy tein relative to the same concentration of resveratrol. A sir droxy on sites 3.5.7.3',4',5'): (-)-epigallocatechin gallate tuin-modulating compound may have an ED50 for modulat (Hydroxy sites 5.7.3',4',5' and gallate ester on 3): cyanidin ing SIRT5 that is at least about 10 fold, 20 fold, 30 fold, 50 chloride (3.5.7.3',4'-pentahydroxyflavylium chloride); del fold greater than that for modulating SIRT1 and/or SIRT3. phinidin chloride (3.5.7.3',4',5'-hexahydroxyflavylium chlo ride); myricetin (cannabiscetin; 3.5.7.3',4',5'-hexahydroxy 3. Exemplary Uses flavone); 3.7.3',4',5'-pentahydroxyflavone; gossypetin (3.5.7. 0239. In certain aspects, the invention provides methods 8.3',4'-hexahydroxyflavone), sirtinol; and splitomicin (see for modulating the leveland/or activity of a sirtuin protein and e.g., Howitz et al. (2003) Nature 425:191: Grozinger et al. methods of use thereof. (2001).J. Biol. Chem. 276:38837; Dedalov et al. (2001) PNAS 0240. In certain embodiments, the invention provides 98: 15113; and Hirao et al. (2003).J. Biol. Chem 278:52773). methods for using sirtuin-modulating compounds wherein In yet another embodiment, one or more sirtuin-modulating the sirtuin-modulating compounds activate a sirtuin protein, compounds may be administered with one or more therapeu e.g., increase the level and/or activity of a sirtuin protein. tic agents for the treatment or prevention of various diseases, Sirtuin-modulating compounds that increase the level and/or including, for example, cancer, diabetes, neurodegenerative activity of a sirtuin protein may be useful for a variety of diseases, cardiovascular disease, blood clotting, inflamma therapeutic applications including, for example, increasing tion, flushing, obesity, ageing, stress, etc. In various embodi the lifespan of a cell, and treating and/or preventing a wide ments, combination therapies comprising a sirtuin-modulat variety of diseases and disorders including, for example, dis ing compound may refer to (1) pharmaceutical compositions eases or disorders related to aging or stress, diabetes, obesity, that comprise one or more sirtuin-modulating compounds in neurodegenerative diseases, cardiovascular disease, blood combination with one or more therapeutic agents (e.g., one or clotting disorders, inflammation, cancer, and/or flushing, etc. more therapeutic agents described herein); and (2) co-admin The methods comprise administering to a Subject in need istration of one or more sirtuin-modulating compounds with thereof a pharmaceutically effective amount of a sirtuin one or more therapeutic agents wherein the sirtuin-modulat modulating compound, e.g., a sirtuin-activating compound. ing compound and therapeutic agent have not been formu 0241. In other embodiments, the invention provides meth lated in the same compositions (but may be present within the ods for using sirtuin-modulating compounds wherein the sir same kit or package. Such as a blister pack or other multi tuin-modulating compounds decrease sirtuin activity, e.g., chamber package; connected, separately sealed containers decrease the level and/or activity of a sirtuin protein. Sirtuin (e.g., foil pouches) that can be separated by the user; or a kit modulating compounds that decrease the level and/or activity where the sirtuin-modulating compound(s) and other thera of a sirtuin protein may be useful for a variety of therapeutic peutic agent(s) are in separate vessels). When using separate application including, for example, increasing cellular sensi formulations, the sirtuin-modulating compound may be tivity to stress (including increasing radiosensitivity and/or administered at the same, intermittent, staggered, prior to, chemosensitivity), increasing the amount and/or rate of apo Subsequent to, or combinations thereof, with the administra ptosis, treatment of cancer (optionally in combination tion of another therapeutic agent. another chemotherapeutic agent), Stimulation of appetite, 0244. In certain embodiments, methods for reducing, pre and/or stimulation of weight gain, etc. The methods comprise venting or treating diseases or disorders using a sirtuin-modu administering to a subject in need thereof a pharmaceutically lating compound may also comprise increasing the protein effective amount of a sirtuin-modulating compound, e.g., a level of a sirtuin, such as human SIRT1 and/or SIRT3, or sirtuin-inhibiting compound. homologs thereof. Increasing protein levels can be achieved 0242. While Applicants do not wish to be bound by theory, by introducing into a cell one or more copies of a nucleic acid it is believed that activators and inhibitors of the instant inven that encodes a sirtuin. For example, the level of a sirtuin can tion may interact with a sirtuin at the same location within the be increased in a mammalian cell by introducing into the sirtuin protein (e.g., active site or site affecting the Kin or mammalian cell a nucleic acid encoding the sirtuin, e.g., Vmax of the active site). It is believed that this is the reason increasing the level of SIRT1 by introducing a nucleic acid why certain classes of sirtuin activators and inhibitors can encoding the amino acid sequence set forth in GenBank have Substantial structural similarity. Accession No. NP 036370 and/or increasing the level of 0243 In certain embodiments, the sirtuin-modulating SIRT3 by introducing a nucleic acid encoding the amino acid compounds described herein may be taken alone or in com sequence set forth in GenBank Accession No. AAHO1042. bination with other compounds. In one embodiment, a mix The nucleic acid may be under the control of a promoter that ture of two or more sirtuin-modulating compounds may be regulates the expression of the SIRT1 and/or SIRT3 nucleic administered to a subject in need thereof. In another embodi acid. Alternatively, the nucleic acid may be introduced into ment, a sirtuin-modulating compound that increases the level the cell at a location in the genome that is downstream of a US 2009/0143376 A1 Jun. 4, 2009

promoter. Methods for increasing the level of a protein using the Survival of a cell, delaying cellular senescence in a cell, these methods are well known in the art. mimicking the effects of calorie restriction, increasing the 0245. A nucleic acid that is introduced into a cell to resistance of a cell to stress, or preventing apoptosis of a cell, increase the protein level of a sirtuin may encode a protein by contacting the cell with a sirtuin-modulating compound of that is at least about 80%, 85%, 90%, 95%, 98%, or 99% the invention that increases the level and/or activity of a identical to the sequence of a sirtuin, e.g., SIRT1 (GenBank sirtuin protein. In an exemplary embodiment, the methods Accession No. NP 036370) and/or SIRT3 (GenBankAcces comprise contacting the cell with a sirtuin-activating com sion No. AAHO1042) protein. For example, the nucleic acid pound. encoding the protein may be at least about 80%, 85%, 90%, 0249. The methods described herein may be used to 95%, 98%, or 99% identical to a nucleic acid encoding a increase the amount of time that cells, particularly primary SIRT1 (e.g. GenBank Accession No. NM 012238) and/or cells (i.e., cells obtained from an organism, e.g., a human), SIRT3 (e.g., GenBank Accession No. BC001042) protein. may be kept alive in a cell culture. Embryonic stem (ES) cells The nucleic acid may also be a nucleic acid that hybridizes, and pluripotent cells, and cells differentiated therefrom, may preferably under Stringent hybridization conditions, to a also be treated with a sirtuin-modulating compound that nucleic acid encoding a wild-type sirtuin, e.g., SIRT1 (Gen increases the level and/or activity of a sirtuin protein to keep Bank Accession No. NM 012238) and/or SIRT3 (e.g., Gen the cells, or progeny thereof, in culture for longer periods of Bank Accession No. BC001042) protein. Stringent hybrid time. Such cells can also be used for transplantation into a ization conditions may include hybridization and a wash in Subject, e.g., after ex vivo modification. 0.2xSSC at 65° C. When using a nucleic acid that encodes a (0250. In one embodiment, cells that are intended to be protein that is different from a wild-type sirtuin protein, such preserved for long periods of time may be treated with a as a protein that is a fragment of a wild-type sirtuin, the sirtuin-modulating compound that increases the level and/or protein is preferably biologically active, e.g., is capable of activity of a sirtuin protein. The cells may be in Suspension deacetylation. It is only necessary to express in a cell a portion (e.g., blood cells, serum, biological growth media, etc.) or in of the sirtuin that is biologically active. For example, a protein tissues or organs. For example, blood collected from an indi that differs from wild-type SIRT1 having GenBank Acces vidual for purposes of transfusion may be treated with a sion No. NP 036370, preferably contains the core structure sirtuin-modulating compound that increases the level and/or thereof. The core structure sometimes refers to amino acids activity of a sirtuin protein to preserve the blood cells for 62-293 of GenBank Accession No. NP 036370, which are longer periods of time. Additionally, blood to be used for encoded by nucleotides 237 to 932 of GenBank Accession forensic purposes may also be preserved using a sirtuin No. NM 012238, which encompasses the NAD binding as modulating compound that increases the level and/or activity well as the substrate binding domains. The core domain of of a sirtuin protein. Other cells that may be treated to extend SIRT1 may also refer to about amino acids 261 to 447 of their lifespan or protect against apoptosis include cells for GenBank Accession No. NP 036370, which are encoded by consumption, e.g., cells from non-human mammals (such as nucleotides 834 to 1394 of GenBank Accession No. meat) or plant cells (such as Vegetables). NM 012238; to about amino acids 242 to 493 of GenBank 0251 Sirtuin-modulating compounds that increase the Accession No. NP 036370, which are encoded by nucle level and/or activity of a sirtuin protein may also be applied otides 777 to 1532 of GenBank Accession No. NM 012238; during developmental and growth phases in mammals, plants, or to aboutamino acids 254 to 495 of GenBank Accession No. insects or microorganisms, in order to, e.g., alter, retard or NP 036370, which are encoded by nucleotides 813 to 1538 accelerate the developmental and/or growth process. of GenBank Accession No. NM 012238. Whether a protein 0252. In another embodiment, sirtuin-modulating com retains a biological function, e.g., deacetylation capabilities, pounds that increase the level and/or activity of a sirtuin can be determined according to methods known in the art. protein may be used to treat cells useful for transplantation or 0246. In certain embodiments, methods for reducing, pre cell therapy, including, for example, Solid tissue grafts, organ venting or treating diseases or disorders using a sirtuin-modu transplants, cell Suspensions, stem cells, bone marrow cells, lating compound may also comprise decreasing the protein etc. The cells or tissue may be an autograft, an allograft, a level of a sirtuin, such as human SIRT1 and/or SIRT3, or Syngraft or a Xenograft. The cells or tissue may be treated with homologs thereof. Decreasing a sirtuin protein level can be the sirtuin-modulating compound prior to administration/im achieved according to methods known in the art. For example, plantation, concurrently with administration/implantation, an siRNA, an antisense nucleic acid, or a ribozyme targeted to and/or post administration/implantation into a Subject. The the sirtuin can be expressed in the cell. A dominant negative cells or tissue may be treated prior to removal of the cells from sirtuin mutant, e.g., a mutant that is not capable of deacety the donor individual, ex vivo after removal of the cells or lating, may also be used. For example, mutant H363Y of tissue from the donor individual, or post implantation into the SIRT1, described, e.g., in Luo et al. (2001) Cell 107:137 can recipient. For example, the donor or recipient individual may be used. Alternatively, agents that inhibit transcription can be be treated Systemically with a sirtuin-modulating compound used. or may have a subset of cells/tissue treated locally with a 0247 Methods for modulating sirtuin protein levels also sirtuin-modulating compound that increases the level and/or include methods for modulating the transcription of genes activity of a sirtuin protein. In certain embodiments, the cells encoding sirtuins, methods for stabilizing/destabilizing the or tissue (or donor/recipient individuals) may additionally be corresponding mRNAS, and other methods known in the art. treated with another therapeutic agent useful for prolonging graft Survival. Such as, for example, an immunosuppressive Aging/Stress agent, a cytokine, an angiogenic factor, etc. 0248. In one embodiment, the invention provides a 0253) In yet other embodiments, cells may be treated with method extending the lifespan of a cell, extending the prolif a sirtuin-modulating compound that increases the leveland/or erative capacity of a cell, slowing ageing of a cell, promoting activity of a sirtuin protein in Vivo, e.g., to increase their US 2009/0143376 A1 Jun. 4, 2009 lifespan or prevent apoptosis. For example, skin can be pro 0258 Sirtuin-modulating compounds that increase the tected from aging (e.g., developing wrinkles, loss of elastic level and/or activity of a sirtuin protein may be administered ity, etc.) by treating skin or epithelial cells with a sirtuin to a subject to prevent aging and aging-related consequences modulating compound that increases the level and/or activity or diseases, such as stroke, heart disease, heart failure, arthri of a sirtuin protein. In an exemplary embodiment, skin is tis, high blood pressure, and Alzheimer's disease. Other con contacted with a pharmaceutical or cosmetic composition ditions that can be treated include ocular disorders, e.g., asso comprising a sirtuin-modulating compound that increases the ciated with the aging of the eye. Such as cataracts, glaucoma, level and/or activity of a sirtuin protein. Exemplary skin and macular degeneration. Sirtuin-modulating compounds afflictions or skin conditions that may be treated in accor that increase the level and/or activity of a sirtuin protein can dance with the methods described herein include disorders or also be administered to Subjects for treatment of diseases, diseases associated with or caused by inflammation, Sundam e.g., chronic diseases, associated with cell death, in order to age or natural aging. For example, the compositions find protect the cells from cell death. Exemplary diseases include utility in the prevention or treatment of contact dermatitis those associated with neural cell death, neuronal dysfunction, (including irritant contact dermatitis and allergic contact der or muscular cell death or dysfunction, such as Parkinson's matitis), atopic dermatitis (also known as allergic eczema), disease, Alzheimer's disease, multiple Sclerosis, amniotropic actinic keratosis, keratinization disorders (including lateral Sclerosis, and muscular dystrophy: AIDS; fulminant eczema), epidermolysis bullosa diseases (including penfi hepatitis; diseases linked to degeneration of the brain, Such as gus), exfoliative dermatitis, seborrheic dermatitis, erythemas Creutzfeld-Jakob disease, retinitis pigmentosa and cerebellar (including erythema multiforme and erythema nodosum), degeneration; myelodysplasis Such as aplastic anemia; damage caused by the Sun or other light sources, discoid lupus ischemic diseases Such as myocardial infarction and stroke; erythematosus, dermatomyositis, psoriasis, skin cancer and hepatic diseases such as alcoholic hepatitis, hepatitis B and the effects of natural aging. In another embodiment, sirtuin hepatitis C; joint-diseases such as osteoarthritis; atheroscle modulating compounds that increase the level and/or activity rosis; alopecia; damage to the skin due to UV light; lichen of a sirtuin protein may be used for the treatment of wounds planus; atrophy of the skin; cataract; and graft rejections. Cell and/or burns to promote healing, including, for example, death can also be caused by Surgery, drug therapy, chemical first-, second- or third-degree burns and/or a thermal, chemi exposure or radiation exposure. cal or electrical burns. The formulations may be administered 0259 Sirtuin-modulating compounds that increase the topically, to the skin or mucosal tissue, as an ointment, lotion, level and/or activity of a sirtuin protein can also be adminis cream, microemulsion, gel, Solution or the like, as further tered to a subject Suffering from an acute disease, e.g., dam described herein, within the context of a dosing regimen age to an organ or tissue, e.g., a subject Suffering from stroke effective to bring about the desired result. or myocardial infarction or a Subject Suffering from a spinal 0254 Topical formulations comprising one or more sir cord injury. Sirtuin-modulating compounds that increase the tuin-modulating compounds that increase the level and/or level and/or activity of a sirtuin protein may also be used to activity of a sirtuin protein may also be used as preventive, repair an alcoholic’s liver. e.g., chemopreventive, compositions. When used in a chemo preventive method, Susceptible skin is treated prior to any Cardiovascular Disease visible condition in a particular individual. 0260. In another embodiment, the invention provides a 0255 Sirtuin-modulating compounds may be delivered method for treating and/or preventing a cardiovascular dis locally or systemically to a Subject. In one embodiment, a ease by administering to a subject in need thereof a sirtuin sirtuin-modulating compound is delivered locally to a tissue modulating compound that increases the level and/or activity or organ of a Subject by injection, topical formulation, etc. of a sirtuin protein. 0256 In another embodiment, a sirtuin-modulating com 0261 Cardiovascular diseases that can be treated or pre pound that increases the level and/or activity of a sirtuin vented using the sirtuin-modulating compounds that increase protein may be used for treating or preventing a disease or the level and/or activity of a sirtuin protein include cardiomy condition induced or exacerbated by cellular senescence in a opathy or myocarditis; such as idiopathic cardiomyopathy, Subject; methods for decreasing the rate of senescence of a metabolic cardiomyopathy, alcoholic cardiomyopathy, drug Subject, e.g., after onset of senescence; methods for extending induced cardiomyopathy, ischemic cardiomyopathy, and the lifespan of a Subject; methods for treating or preventing a hypertensive cardiomyopathy. Also treatable or preventable disease or condition relating to lifespan; methods for treating using compounds and methods described herein are athero or preventing a disease or condition relating to the prolifera matous disorders of the major blood vessels (macrovascular tive capacity of cells; and methods for treating or preventing disease) Such as the aorta, the coronary arteries, the carotid a disease or condition resulting from cell damage or death. In arteries, the cerebrovascular arteries, the renal arteries, the certain embodiments, the method does not act by decreasing iliac arteries, the femoral arteries, and the popliteal arteries. the rate of occurrence of diseases that shorten the lifespan of Other vascular diseases that can be treated or prevented a subject. In certain embodiments, a method does not act by include those related to platelet aggregation, the retinal arte reducing the lethality caused by a disease, such as cancer. rioles, the glomerular arterioles, the Vasa nervorum, cardiac 0257. In yet another embodiment, a sirtuin-modulating arterioles, and associated capillary beds of the eye, the kidney, compound that increases the level and/or activity of a sirtuin the heart, and the central and peripheral nervous systems. The protein may be administered to a Subject in order to generally sirtuin-modulating compounds that increase the level and/or increase the lifespan of its cells and to protect its cells against activity of a sirtuin protein may also be used for increasing stress and/or against apoptosis. It is believed that treating a HDL levels in plasma of an individual. subject with a compound described herein is similar to sub 0262 Yet other disorders that may be treated with sirtuin jecting the Subject to hormesis, i.e., mild stress that is benefi modulating compounds that increase the level and/or activity cial to organisms and may extend their lifespan. of a sirtuin protein include restenosis, e.g., following coro US 2009/0143376 A1 Jun. 4, 2009

nary intervention, and disorders relating to an abnormal level activator complex (APSAC, Eminase, Beecham Laborato of high density and low density cholesterol. ries), or animal salivary gland plasminogen activators, cal 0263. In one embodiment, a sirtuin-modulating com cium channel blocking agents such as Verapamil, nifedipine pound that increases the level and/or activity of a sirtuin or diltiazem, thromboxane receptor antagonists Such as protein may be administered as part of a combination thera ifetroban, prostacyclin mimetics, or phosphodiesterase peutic with another cardiovascular agent including, for inhibitors. Such combination products if formulated as a fixed example, an anti-arrhythmic agent, an antihypertensive dose employ the compounds of this invention within the dose agent, a calcium channel blocker, a cardioplegic Solution, a range described above and the other pharmaceutically active cardiotonic agent, a fibrinolytic agent, a Sclerosing solution, a agent within its approved dose range. vasoconstrictor agent, a vasodilator agent, a nitric oxide 0267 Yet other exemplary cardiovascular agents include, donor, a potassium channel blocker, a sodium channel for example, vasodilators, e.g., bencyclane, cinnarizine, citi blocker, statins, or a naturiuretic agent. coline, cyclandelate, cyclonicate, ebumamonine, phenox 0264. In one embodiment, a sirtuin-modulating com eZyl, flunarizine, ibudilast, ifenprodil, lomerizine, naphlole, pound that increases the level and/or activity of a sirtuin nikamate, nosergoline, nimodipine, papaverine, pentifylline, protein may be administered as part of a combination thera nofedoline, Vincamin, Vinpocetine, Vichizyl, pentoxifylline, peutic with an anti-arrhythmia agent. Anti-arrhythmia agents prostacyclin derivatives (such as prostaglandin E1 and pros are often organized into four main groups according to their taglandin I2), an endothelin receptor blocking drug (Such as mechanism of action: type I, Sodium channel blockade; type bosentan), diltiazem, nicorandil, and nitroglycerin. Examples II, beta-adrenergic blockade; type I, repolarization prolonga of the cerebral protecting drug include radical scavengers tion; and type IV, calcium channel blockade. Type I anti (such as edaravone, Vitamin E, and vitamin C), glutamate arrhythmic agents include lidocaine, moricizine, mexiletine, antagonists, AMPA antagonists, kainate antagonists, NMDA tocainide, procainamide, encainide, flecanide, tocainide, phe antagonists, GABA agonists, growth factors, opioid antago nyloin, propafenone, quinidine, disopyramide, and flecain nists, phosphatidylcholine precursors, serotonin agonists, ide. Type II anti-arrhythmic agents include propranolol and Na"/Ca" channel inhibitory drugs, and K" channel opening esmolol. Type III includes agents that act by prolonging the drugs. Examples of the brain metabolic stimulants include duration of the action potential. Such as amiodarone, artilide, amantadine, tiapride, and gamma.-aminobutyric acid. bretylium, clofilium, isobutilide, Sotalol, azimilide, dofetil Examples of the anticoagulant include heparins (such as hep ide, dronedarone, ersentilide, ibutilide, tedisamil, and tre arin Sodium, heparin potassium, dalteparin Sodium, dalte cetilide. Type IV anti-arrhythmic agents include Verapamil, parin calcium, heparin calcium, parnaparin Sodium, reviparin dilitaizem, digitalis, adenosine, nickel chloride, and magne Sodium, and danaparoid sodium), warfarin, enoxaparin, arga sium ions. troban, batroXobin, and Sodium citrate. Examples of the anti 0265. In another embodiment, a sirtuin-modulating com platelet drug include ticlopidine hydrochloride, dipy pound that increases the level and/or activity of a sirtuin ridamole, ciloStaZol, ethyl icosapentate, Sarpogrelate protein may be administered as part of a combination thera hydrochloride, dilazep hydrochloride, trapidil, a nonsteroidal peutic with another cardiovascular agent. Examples of car antiinflammatory agent (such as aspirin), beraprostSodium, diovascular agents include vasodilators, for example, iloprost, and indobufene. Examples of the thrombolytic drug hydralazine; angiotensin converting enzyme inhibitors, for include urokinase, tissue-type plasminogen activators (such example, captopril; anti-anginal agents, for example, isosor as alteplase, tisokinase, nateplase, pamiteplase, monteplase, bide nitrate, glyceryl trinitrate and pentaerythritol tetrani and rateplase), and nasaruplase. Examples of the antihyper trate; anti-arrhythmic agents, for example, quinidine, tensive drug include angiotensin converting enzyme inhibi procainaltide and lignocaine; cardioglycosides, for example, tors (such as captopril, alacepril, lisinopril, imidapril, digoxin and digitoxin; calcium antagonists, for example, quinapril, temocapril, delapril, benazepril, cilaZapril, tran Verapamil and nifedipine; diuretics, such as thiazides and dollapril, enalapril, ceronapril, fosinopril, imadapril, mobert related compounds, for example, bendrofluazide, chlorothi pril, perindopril, ramipril, spirapril, and randolapril), angio azide, chlorothalidone, hydrochlorothiazide and other diuret tensin II antagonists (such as losartan, candesartan, Valsartan, ics, for example, fursemide and triamterene, and sedatives, eprosartan, and irbesartan), calcium channel blocking drugs for example, nitrazepam, flurazepam and diazepam. (such as aranidipine, efonidipine, nicardipine, bamidipine, 0266 Other exemplary cardiovascular agents include, for benidipine, manidipine, cilnidipine, niSoldipine, nitren example, a cyclooxygenase inhibitor Such as aspirin or dipine, nifedipine, nilvadipine, felodipine, amlodipine, dilt indomethacin, a platelet aggregation inhibitor Such as clopi iazem, bepridil, clentiaZem, phendilin, galopamil, milbe dogrel, ticlopidene or aspirin, fibrinogen antagonists or a fradil, prenylamine, semotiadil, terodiline, Verapamil, diuretic such as chlorothiazide, hydrochlorothiazide, flume cilnidipine, elgodipine, isradipine, lacidipine, lercanidipine, thiazide, hydroflumethiazide, bendroflumethiazide, methyl nimodipine, cinnarizine, flunarizine, lidoflazine, lomerizine, chlorthiazide, trichloromethiazide, polythiazide or benzthi bencyclane, etafenone, and perhexyline), B-adrenaline recep azide as well as ethacrynic acid tricrynafen, chlorthalidone, tor blocking drugs (propranolol, pindolol, indenolol, car furosemide, musolimine, bumetanide, triamterene, amiloride teolol, bunitrolol, atenolol, acebutolol, metoprolol, timolol, and Spironolactone and salts of such compounds, angiotensin nipradillol, penbutolol, nadolol, tilisolol, carvedilol, biso converting enzyme inhibitors such as captopril. Zofenopril, prolol, betaxolol, celiprolol, bopindolol, bevantolol, labe fosinopril, enalapril, ceranopril, cilaZopril, delapril, pento talol, alprenolol, amoSulalol, arotinolol, befunolol, bucu pril, quinapril, ramipril, lisinopril, and salts of Such com molol, bufetolol, buferalol, buprandolol, butylidine, pounds, angiotensin II antagonists such as losartan, irbesartan butofilolol, carazolol, cetamolol, cloranolol, dilevalol, or Valsartan, thrombolytic agents such as tissue plasminogen epanolol, levobunolol, mepindolol, metipranolol, moprolol. activator (tPA), recombinant tRA, streptokinase, urokinase, nadoxolol, nevibolol, Oxprenolol, practol, pronetalol, Sotalol, prourokinase, and anisoylated plasminogen streptokinase Sufinalol, talindolol, tertalol, toliprolol, xybenolol, and US 2009/0143376 A1 Jun. 4, 2009

esmolol), C.-receptor blocking drugs (such as amoSulalol, a dose of radiation or toxin. In one embodiment, the dose of prazosin, teraZosin, doxazosin, bunaZosin, urapidil, phento radiation or toxin is received as part of a work-related or lamine, arotinolol, dapiprazole, fenspiride, indoramin, labe medical procedure, e.g., working in a nuclear power plant, talol, naftopidil, nicergoline, tamsulosin, tolazoline, trima flying an airplane, an X-ray, CAT scan, or the administration Zosin, and yohimbine), sympathetic nerve inhibitors (such as of a radioactive dye for medical imaging; in Such an embodi clonidine, guanfacine, guanabenz, methyldopa, and reser ment, the compound is administered as a prophylactic mea pine), hydralazine, todralazine, budralazine, and cadralazine. Sure. In another embodiment, the radiation or toxin exposure Examples of the antianginal drug include nitrate drugs (such is received unintentionally, e.g., as a result of an industrial as amyl nitrite, nitroglycerin, and isosorbide), B-adrenaline accident, habitation in a location of natural radiation, terrorist receptor blocking drugs (such as propranolol, pindolol, inde act, or act of war involving radioactive or toxic material. In nolol, carteolol, bunitrolol, atenolol, acebutolol, metoprolol. Such a case, the compound is preferably administered as soon timolol, nipradillol, penbutolol, nadolol, tilisolol, carvedilol. as possible after the exposure to inhibit apoptosis and the bisoprolol, betaxolol, celiprolol, bopindolol, bevantolol. Subsequent development of acute radiation syndrome. labetalol, alprenolol, amosulalol, arotinolol, befunolol, bucu 0269 Sirtuin-modulating compounds may also be used molol, bufetolol, buferalol, buprandolol, butylidine, buto for treating and/or preventing cancer. In certain embodi filolol, carazolol, cetamolol, cloranolol, dilevalol, epanolol. ments, sirtuin-modulating compounds that increase the level levobunolol, mepindolol, metipranolol, moprolol, nadoxolol. and/or activity of a sirtuin protein may be used for treating nevibolol, oXprenolol, practol, pronetalol, Sotalol, Sufinalol, and/or preventing cancer. Calorie restriction has been linked talindolol, tertalol, toliprolol, andxybenolol), calcium chan to a reduction in the incidence of age-related disorders includ nel blocking drugs (such as aranidipine, efonidipine, nicar ing cancer (see e.g., Bordone and Guarente, Nat. Rev. Mol. dipine, bamidipine, benidipine, manidipine, cilnidipine, Cell. Biol. (2005 epub): Guarente and Picard, Cell 120: 473 nisoldipine, nitrendipine, nifedipine, nilvadipine, felodipine, 82 (2005); Berrigan, et al., Carcinogenesis 23: 817-822 amlodipine, diltiazem, bepridil, clentiazem, phendiline, galo (2002); and Heilbronn and Ravussin, Am. J. Clin. Nutr. 78: pamil, mibefradil, prenylamine, Semotiadil, terodiline, Vera 361-369 (2003)). Additionally, the Sir2 protein from yeast pamil, cilnidipine, elgodipine, isradipine, lacidipine, lercani has been shown to be required for lifespan extension by dipine, nimodipine, cinnarizine, flunarizine, lidoflazine, glucose restriction (see e.g., Lin et al., Science 289: 2126 lomerizine, bencyclane, etafenone, and perhexyline) tri 2128 (2000); Anderson et al., Nature 423: 181-185 (2003)), a metazidine, dipyridamole, etafenone, dilaZep, trapidil, nic yeast model for calorie restriction. Accordingly, an increase orandil, enoxaparin, and aspirin. Examples of the diuretic in the level and/or activity of a sirtuin protein may be useful include thiazide diuretics (such as hydrochlorothiazide, for treating and/or preventing the incidence of age-related methyclothiazide, trichloromethiazide, benzylhydrochlo disorders, such as, for example, cancer. In other embodi rothiazide, and penflutizide), loop diuretics (such as furo ments, sirtuin-modulating compounds that decrease the level semide, etacrynic acid, bumetanide, piretanide, azosemide, and/or activity of a sirtuin protein may be used for treating or and torasemide), K" sparing diuretics (spironolactone, triam preventing cancer. For example, inhibitory compounds may terene, and potassiumcanrenoate), osmotic diuretics (such as be used to stimulate acetylation of substrates such as p53 and isosorbide, D-mannitol, and glycerin), nonthiazide diuretics thereby increase apoptosis, as well as to reduce the lifespan of (such as meticrane, tripamide, chlorthalidone, and mefru cells and organisms, render them more sensitive to stress, side), and acetazolamide. Examples of the cardiotonic and/or increase the radiosensitivity and/or chemosensitivity include digitalis formulations (such as digitoxin, digoxin, of a cell or organism. Thus, inhibitory compounds may be methyldigoxin, deslanoside, Vesnarinone, lanatoside C, and used, e.g., for treating cancer. Exemplary cancers that may be proscillaridin), Xanthine formulations (such as aminophyl treated using a sirtuin-modulating compound are those of the line, choline theophylline, diprophylline, and proxyphylline), brain and kidney; hormone-dependent cancers including catecholamine formulations (such as dopamine, dobutamine, breast, prostate, testicular, and ovarian cancers; lymphomas, and docarpamine), PDE III inhibitors (such as amrinone, and leukemias. In cancers associated with solid tumors, a olprinone, and milrinone), denopamine, ubidecarenone, modulating compound may be administered directly into the pimobendan, levosimendan, aminoethylsulfonic acid, tumor. Cancer of blood cells, e.g., leukemia, can be treated by Vesnarinone, carperitide, and colforsin daropate. Examples of administering a modulating compound into the blood stream the antiarrhythmic drug include ajmaline, pirmenol, or into the bone marrow. Benign cell growth can also be procainamide, cibenzoline, disopyramide, quinidine, aprind treated, e.g., warts. Other diseases that can be treated include ine, mexiletine, lidocaine, phenyloin, pilsicainide, pro autoimmune diseases, e.g., systemic lupus erythematosus, pafenone, flecainide, atenolol, acebutolol, Sotalol, propra Scleroderma, and arthritis, in which autoimmune cells should nolol, metoprolol, pindolol, amiodarone, nifekalant, be removed. Viral infections such as herpes, HIV, adenovirus, diltiazem, bepridil, and Verapamil. Examples of the antihy and HTLV-1 associated malignant and benign disorders can perlipidemic drug include atorvastatin, simvastatin, pravas also be treated by administration of sirtuin-modulating com tatin Sodium, fluvastatin Sodium, clinofibrate, clofibrate, sim pound. Alternatively, cells can be obtained from a Subject, fibrate, fenofibrate, bezafibrate, colestimide, and treated ex vivo to remove certain undesirable cells, e.g., can colestyramine. Examples of the immunosuppressant include cer cells, and administered back to the same or a different azathioprine, mizoribine, cyclosporine, tacrolimus, gusperi Subject. mus, and methotrexate. 0270 Chemotherapeutic agents that may be coadminis tered with modulating compounds described herein as having Cell Death/Cancer anti-cancer activity (e.g., compounds that induce apoptosis, 0268 Sirtuin-modulating compounds that increase the compounds that reduce lifespan or compounds that render level and/or activity of a sirtuin protein may be administered cells sensitive to stress) include: aminoglutethimide, amsa to subjects who have recently received or are likely to receive crine, anastroZole, asparaginase, bcg, bicalutamide, bleomy US 2009/0143376 A1 Jun. 4, 2009 20 cin, buserelin, buSulfan, campothecin, capecitabine, carbopl L-asparagine and deprives cells which do not have the capac atin, carmustine, chlorambucil, cisplatin, cladribine, ity to synthesize their own asparagine); antiplatelet agents; clodronate, colchicine, cyclophosphamide, cyproterone, cyt antiproliferative/antimitotic alkylating agents such as nitro arabine, dacarbazine, dactinomycin, daunorubicin, dienes gen mustards (mechlorethamine, cyclophosphamide and ana trol, diethylstilbestrol, docetaxel, doxorabicin, epirubicin, logs, melphalan, chlorambucil), ethylenimines and meth estradiol, estramustine, etoposide, exemestane, filgrastim, ylmelamines (hexamethylmelamine and thiotepa), alkyl fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, sulfonates-busulfan, nitrosoureas (carmustine (BCNU) and flutamide, gemcitabine, genistein, goserelin, hydroxyurea, analogs, streptozocin), traZenes-dacarbazinine (DTIC); anti idarubicin, ifosfamide, imatinib, interferon, irinotecan, proliferative/antimitotic antimetabolites such as folic acid ironotecan, letrozole, leucovorin, leuprolide, levamisole, analogs (methotrexate); platinum coordination complexes lomustine, mechlorethamine, medroxyprogesterone, mege (cisplatin, carboplatin), procarbazine, hydroxyurea, mito strol, melphalan, mercaptopurine, mesna, methotrexate, tane, aminoglutethimide; hormones, hormone analogs (estro mitomycin, mitotane, mitoxantrone, nilutamide, nocodazole, gen, tamoxifen, goserelin, bicalutamide, nilutamide) and aro octreotide, oxaliplatin, paclitaxel, pamidronate, pentostatin, matase inhibitors (letrozole, anastrozole); anticoagulants plicamycin, porfimer, procarbazine, raltitrexed, rituximab, (heparin, synthetic heparin salts and other inhibitors of streptozocin, Suramin, tamoxifen, temozolomide, teniposide, thrombin); fibrinolytic agents (such as tissue plasminogen testosterone, thioguanine, thiotepa, titanocene dichloride, activator, streptokinase and urokinase), aspirin, COX-2 topotecan, trastuzumab, tretinoin, vinblastine, Vincristine, inhibitors, dipyridamole, ticlopidine, clopidogrel, abcix vindesine, and vinorelbine. imab; antimigratory agents; antisecretory agents (breveldin); 0271 These chemotherapeutic agents may be categorized immunosuppressives (cyclosporine, tacrolimus (FK-506), by their mechanism of action into, for example, following Sirolimus (rapamycin), azathioprine, mycophenolate groups: anti-metabolites/anti-cancer agents, such as pyrimi mofetil); anti-angiogenic compounds (TNP-470, genistein) dine analogs (5-fluorouracil, floXuridine, capecitabine, gem and growth factor inhibitors (vascular endothelial growth fac citabine and cytarabine) and purine analogs, folate antago tor (VEGF) inhibitors, fibroblast growth factor (FGF) inhibi nists and related inhibitors (mercaptopurine, thioguanine, tors, epidermal growth factor (EGF) inhibitors); angiotensin pentostatin and 2-chlorodeoxyadenosine (cladribine)); anti receptor blocker, nitric oxide donors; anti-sense oligonucle proliferative/antimitotic agents including natural products otides; antibodies (trastuzumab); cell cycle inhibitors and Such as Vinca alkaloids (vinblastine, Vincristine, and vinorel differentiation inducers (tretinoin); mTOR inhibitors, topoi bine), microtubule disruptors such as taxane (paclitaxel, doc Somerase inhibitors (doxorubicin (adriamycin), amsacrine, etaxel), Vincristin, vinblastin, nocodazole, epothilones and camptothecin, daunorubicin, dactinomycin, eniposide, epiru navelbine, epidipodophyllotoxins (teniposide), DNA damag bicin, etoposide, idarubicin, irinotecan (CPT-11) and mitox ing agents (actinomycin, amsacrine, anthracyclines, bleomy antrone, topotecan, irinotecan), corticosteroids (cortisone, cin, buSulfan, camptothecin, carboplatin, chlorambucil, cis dexamethasone, hydrocortisone, methylpednisolone, pred platin, cyclophosphamide, cytoxan, dactinomycin, nisone, and prenisolone); growth factor signal transduction daunorubicin, docetaxel, doxorubicin, epirubicin, hexameth kinase inhibitors; mitochondrial dysfunction inducers and ylmelamineoxaliplatin, iphosphamide, melphalan, merchlo caspase activators; chromatin disruptors. retlamine, mitomycin, mitoxantrone, nitrosourea, paclitaxel, 0272. These chemotherapeutic agents may be used by plicamycin, procarbazine, teniposide, triethylenethiophos themselves with a sirtuin-modulating compound described phoramide and etoposide (VP16)); antibiotics such as dacti herein as inducing cell death or reducing lifespan or increas nomycin (actinomycin D), daunorubicin, doxorubicin (adria ing sensitivity to stress and/or in combination with other mycin), idarubicin, anthracyclines, mitoxantrone, chemotherapeutics agents. Many combinatorial therapies bleomycins, plicamycin (mithramycin) and mitomycin; have been developed, including but not limited to those listed enzymes (L-asparaginase which systemically metabolizes in Table 1.

TABLE 1. Exemplary combinatorial therapies for the treatment of cancer. Name Therapeutic agents ABV Doxorubicin, Bleomycin, Vinblastine ABVD Doxorubicin, Bleomycin, Vinblastine, Dacarbazine AC (Breast) Doxorubicin, Cyclophosphamide AC (Sarcoma) Doxorubicin, Cisplatin AC (Neuroblastoma) Cyclophosphamide, Doxorubicin ACE Cyclophosphamide, Doxorubicin, Etoposide ACe Cyclophosphamide, Doxorubicin AD Doxorubicin, Dacarbazine AP Doxorubicin, Cisplatin ARAC-DNR Cytarabine, Daunorubicin B-CAWe Bleomycin, Lomustine, Doxorubicin, Vinblastine BCVPP Carmustine, Cyclophosphamide, Vinblastine, Procarbazine, Prednisone BEACOPP Bleomycin, Etoposide, Doxorubicin, Cyclophosphamide, Vincristine, Procarbazine, Prednisone, Filgrastim BEP Bleomycin, Etoposide, Cisplatin BIP Bleomycin, Cisplatin, Ifosfamide, Mesna US 2009/0143376 A1 Jun. 4, 2009 21

TABLE 1-continued Exemplary combinatorial therapies for the treatment of cancer. Name Therapeutic agents BOMP Bleomycin, Vincristine, Cisplatin, Mitomycin CA Cytarabine, Asparaginase CABO Cisplatin, Methotrexate, Bleomycin, Vincristine CAF Cyclophosphamide, Doxorubicin, Fluorouracil CAL-G Cyclophosphamide, Daunorubicin, Vincristine, Prednisone, Asparaginase CAMP Cyclophosphamide, Doxorubicin, Methotrexate, Procarbazine CAP phosphamide, Doxorubicin, Cisplatin CaT platin, Paclitaxel CAV phosphamide, Doxorubicin, Vincristine CAVE ADD CA-VP16 phamide, Doxorubicin, Etoposide CC phamide, Carboplatin CDDPVP-16 Cisplatin, Etoposide CEF phamide, Epirubicin, Fluorouracil CEPP(B) phamide, Etoposide, Prednisone, with or without CEV Cyclophosphamide, Etoposide, Vincristine CF Cisplatin, Fluorouracil or Carboplatin Fluorouracil CHAP Cyclophosphamide or Cyclophosphamide, Altretamine, Doxorubicin, Cisplatin ChIVPP Chlorambucil, Vinblastine, Procarbazine, Prednisone CHOP Cyclophosphamide, Doxorubicin, Vincristine, Prednisone CHOP-BLEO Add Bleomycin to CHOP CISCA ophosphamide, Doxorubicin, Cisplatin CLD-BOMP Bleomycin, Cisplatin, Vincristine, Mitomycin CMF Methotrexate, Fluorouracil, Cyclophosphamide CMFP Cyclophosphamide, Methotrexate, Fluorouracil, Prednisone CMFVP Cyclophosphamide, Methotrexate, Fluorouracil, Vincristine, Prednisone CMV Cisplatin, Methotrexate, Vinblastine CNF Cyclophosphamide, Mitoxantrone, Fluorouracil CNOP Cyclophosphamide, Mitoxantrone, Vincristine, Prednisone COB Cisplatin, Vincristine, Bleomycin CODE Cisplatin, Vincristine, Doxorubicin, Etoposide COMLA Cyclophosphamide, Vincristine, Methotrexate, Leucovorin, Cytarabine COMP hosphamide, Vincristine, Methotrexate, Prednisone Cooper Regimen hosphamide, Methotrexate, Fluorouracil, Vincristine, Prednisone COP phosphamide, Vincristine, Prednisone COPE phosphamide, Vincristine, Cisplatin, Etoposide COPP Cyclophosphamide, Vincristine, Procarbazine, Prednisone CP(Chronic lymphocytic Chlorambucil, Prednisone leukemia) CP (Ovarian Cancer) Cyclophosphamide, Cisplatin CT Cisplatin, Paclitaxel CVD Cisplatin, Vinblastine, Dacarbazine CV Carboplatin, Etoposide, Ifosfamide, Mesna CVP Cyclophosphamide, Vincristine, Prednisome CVPP Lomustine, Procarbazine, Prednisone CYVADIC Cyclophosphamide, Vincristine, Doxorubicin, Dacarbazine DA Daunorubicin, Cytarabine DAT Daunorubicin, Cytarabine, Thioguanine DAV Daunorubicin, Cytarabine, Etoposide DCT Daunorubicin, Cytarabine, Thioguanine DHAP Cisplatin, Cytarabine, Dexamethasone DI Doxorubicin, Ifosfamide DTIC, Tamoxifen Dacarbazine, Tamoxifen DVP Daunorubicin, Vincristine, Prednisone EAP Etoposide, Doxorubicin, Cisplatin EC Etoposide, Carboplatin EFP Etoposie, Fluorouracil, Cisplatin ELF Etoposide, Leucovorin, Fluorouracil EMA 86 Mitoxantrone, Etoposide, Cytarabine EP Etoposide, Cisplatin EVA Etoposide, Vinblastine FAC Fluorouracil, Doxorubicin, Cyclophosphamide FAM Fluorouracil, Doxorubicin, Mitomycin FAMTX Methotrexate, Leucovorin, Doxorubicin FAP Fluorouracil, Doxorubicin, Cisplatin US 2009/0143376 A1 Jun. 4, 2009 22

TABLE 1-continued Exemplary combinatorial therapies for the treatment of cancer. Name Therapeutic agents Fluorouracil, Leucovorin FEC Fluorouracil, Cyclophosphamide, Epirubicin FED Fluorouracil, Etoposide, Cisplatin FL Flutamide, Leuprolide FZ Flutamide, Goserelin acetate implant HDMTX Methotrexate, Leucovorin Hexa-CAF Altretamine, Cyclophosphamide, Methotrexate, Fluorouracil ICE-T Ifosfamide, Carboplatin, Etoposide, Paclitaxel, Mesna IDMTXF6-MP Methotrexate, Mercaptopurine, Leucovorin IE Ifosfamide, Etoposie, Mesna IfoVP Ifosfamide, Etoposide, Mesna IPA Ifosfamide, Cisplatin, Doxorubicin M-2 Vincristine, Carmustine, Cyclophosphamide, Prednisone, Melphalan MAC-III Methotrexate, Leucovorin, Dactinomycin, Cyclophosphamide MACC Methotrexate, Doxorubicin, Cyclophosphamide, Lomustine MACOP-B Methotrexate, Leucovorin, Doxorubicin, Cyclophosphamide, Vincristine, Bleomycin, Prednisone MAID Mesna, Doxorubicin, Ifosfamide, Dacarbazine m-BACOD Bleomycin, Doxorubicin, Cyclophosphamide, Vincristine, Dexamethasone, Methotrexate, Leucovorin MBC Methotrexate, Bleomycin, Cisplatin MC Mitoxantrone, Cytarabine Methotrexate, Fluorouracil, Leucovorin MICE Ifosfamide, Carboplatin, Etoposide, Mesna MINE Mesna, Ifosfamide, Mitoxantrone, Etoposide mini-BEAM Carmustine, Etoposide, Cytarabine, Melphalan MOBP Bleomycin, Vincristine, Cisplatin, Mitomycin MOP Mechlorethamine, Vincristine, Procarbazine MOPP Mechlorethamine, Vincristine, Procarbazine, Prednisone MOPPABV Mechlorethamine, Vincristine, Procarbazine, Prednisone, Doxorubicin, Bleomycin, Vinblastine MP (multiple myeloma) Melphalan, Prednisone MP (prostate cancer) Mitoxantrone, Prednisone MTXF6-MO Methotrexate, Mercaptopurine MTX6-MPVP Methotrexate, Mercaptopurine, Vincristine, Prednisone MTX-CDDPAdr Methotrexate, Leucovorin, Cisplatin, Doxorubicin MV (breast cancer) Mitomycin, Vinblastine MV (acute myelocytic Mitoxantrone, Etoposide leukemia) M-WAC Methotrexate Vinblastine, Doxorubicin, Cisplatin MVP Mitomycin Vinblastine, Cisplatin MVPP Mechlorethamine, Vinblastine, Procarbazine, Prednisone NFL Mitoxantrone, Fluorouracil, Leucovorin NOVP Mitoxantrone, Vinblastine, Vincristine OPA Vincristine, Prednisone, Doxorubicin OPPA Add Procarbazine to OPA. PAC Cisplatin, Doxorubicin PAC-I Cisplatin, Doxorubicin, Cyclophosphamide PA-CI Cisplatin, Doxorubicin PC Paclitaxel, Carboplatin or Paclitaxel, Cisplatin PCV Lomustine, Procarbazine, Vincristine PE Paclitaxel, Estramustine PFL Cisplatin, Fluorouracil, Leucovorin POC Prednisone, Vincristine, Lomustine ProMACE Prednisone, Methotrexate, Leucovorin, Doxorubicin, Cyclophosphamide, Etoposide ProMACE cytaBOM Prednisone, Doxorubicin, Cyclophosphamide, Etoposide, Cytarabine, Bleomycin, Vincristine, Methotrexate, Leucovorin, Cotrimoxazole PROMACEMOPP Prednisone, Doxorubicin, Cyclophosphamide, Etoposide, Mechlorethamine, Vincristine, Procarbazine, Methotrexate, Leucovorin Cisplatin, Teniposide Prednisone, Vincristine, Asparaginase Cisplatin, Vinblastine, Bleomycin Prednisone, Vincristine, Daunorubicin, Asparaginase Streptozocin, Mitomycin, Fluorouracil Mechlorethamine, Doxorubicin, Vinblastine, Vincristine, Bleomycin, Etoposide, Prednisone US 2009/0143376 A1 Jun. 4, 2009

TABLE 1-continued Exemplary combinatorial therapies for the treatment of cancer. Name Therapeutic agents TCF Paclitaxel, Cisplatin, Fluorouracil TIP Paclitaxel, Ifosfamide, Mesna, Cisplatin TTT Methotrexate, Cytarabine, Hydrocortisone Topo/CTX Cyclophosphamide, Topotecan, MeSna VAB-6 Cyclophosphamide, Dactinomycin, Vinblastine, Cisplatin, Bleomycin WAC Vincristine, Dactinomycin, Cyclophosphamide WACAdr Vincristine, Cyclophosphamide, Doxorubicin, Dactinomycin, Vincristine WAD Vincristine, Doxorubicin, Dexamethasone WATH Vinblastine, Doxorubicin, Thiotepa, Flouxymesterone WBAP Vincristine, Carmustine, Doxorubicin, Prednisone VBCMP Vincristine, Carmustine, Melphalan, Cyclophosphamide, Prednisone VC Vinorelbine, Cisplatin VCAP Vincristine, Cyclophosphamide, Doxorubicin, Prednisone VD Vinorelbine, Doxorubicin WeIP Vinblastine, Cisplatin, Ifosfamide, Mesna VIP Etoposide, Cisplatin, Ifosfamide, Mesna VM Mitomycin, Vinblastine VMCP Vincristine, Melphalan, Cyclophosphamide, Prednisone VP Etoposide, Cisplatin WTAD Etoposide, Thioguanine, Daunorubicin, Cytarabine 5 - 2 Cytarabine, Daunorubicin, Mitoxantrone 7 - 3 Cytarabine with, Daunorubicin or Idarubicin or Mitoxantrone 8 in 1 Methylprednisolone, Vincristine, Lomustine, Procarbazine, Hydroxyurea, Cisplatin, Cytarabine, Dacarbazine

0273. In addition to conventional chemotherapeutics, the system (PNS). Neurodegenerative disease typically involves sirtuin-modulating compounds described hereinas capable of reductions in the mass and Volume of the human brain, which inducing cell death or reducing lifespan can also be used with may be due to the atrophy and/or death of brain cells, which antisense RNA, RNAi or other polynucleotides to inhibit the are far more profound than those in a healthy person that are expression of the cellular components that contribute to attributable to aging. Neurodegenerative diseases can evolve unwanted cellular proliferation that are targets of conven gradually, after a long period of normal brain function, due to tional chemotherapy. Such targets are, merely to illustrate, progressive degeneration (e.g., nerve cell dysfunction and growth factors, growth factor receptors, cell cycle regulatory death) of specific brain regions. Alternatively, neurodegen proteins, transcription factors, or signal transduction kinases. erative diseases can have a quick onset, Such as those associ 0274 Combination therapies comprising sirtuin-modulat ated with trauma or toxins. The actual onset of brain degen ing compounds and a conventional chemotherapeutic agent eration may precede clinical expression by many years. may be advantageous over combination therapies known in the art because the combination allows the conventional che Examples of neurodegenerative diseases include, but are not motherapeutic agent to exert greater effect at lower dosage. In limited to, Alzheimer's disease (AD), Parkinson's disease a preferred embodiment, the effective dose (EDs) for a che (PD), Huntington's disease (HD), amyotrophic lateral scle motherapeutic agent, or combination of conventional chemo rosis (ALS: Lou Gehrig's disease), diffuse Lewy body dis therapeutic agents, when used in combination with a sirtuin ease, chorea-acanthocytosis, primary lateral Sclerosis, ocular modulating compound is at least 2 fold less than the EDso for diseases (ocular neuritis), chemotherapy-induced neuropa the chemotherapeutic agent alone, and even more preferably thies (e.g., from Vincristine, paclitaxel, bortezomib), diabe at 5 fold, 10 fold or even 25 fold less. Conversely, the thera tes-induced neuropathies and Friedreich's ataxia. Sirtuin peutic index (TI) for Such chemotherapeutic agent or combi modulating compounds that increase the level and/or activity nation of Such chemotherapeutic agent when used in combi of a sirtuin protein can be used to treat these disorders and nation with a sirtuin-modulating compound described herein others as described below. can be at least 2 fold greater than the TI for conventional 0276 AD is a chronic, incurable, and unstoppable CNS chemotherapeutic regimen alone, and even more preferably disorder that occurs gradually, resulting in memory loss, at 5 fold, 10 fold or even 25 fold greater. unusual behavior, personality changes, and a decline in think ing abilities. These losses are related to the death of specific Neuronal Diseases/Disorders types of brain cells and the breakdown of connections and their Supporting network (e.g. glial cells) between them. AD 0275. In certain aspects, sirtuin-modulating compounds has been described as childhood development in reverse. In that increase the level and/or activity of a sirtuin protein can most people with AD, symptoms appear after the age 60. The be used to treat patients suffering from neurodegenerative earliest symptoms include loss of recent memory, faulty judg diseases, and traumatic or mechanical injury to the central ment, and changes in personality. Later in the disease, those nervous system (CNS), spinal cord or peripheral nervous with AD may forget how to do simple tasks like washing their US 2009/0143376 A1 Jun. 4, 2009 24 hands. Eventually people with AD lose all reasoning abilities begins in early infancy. A precipitous neurodegenerative and become dependent on other people for their everyday course then ensues, with affected infants exhibiting motor care. Finally, the disease becomes so debilitating that patients dysfunction, seizure, visual loss, and deafness. Death usually are bedridden and typically develop coexisting illnesses. occurs by 2-5 years of age. Neuronal loss through an apop 0277 PD is a chronic, incurable, and unstoppable CNS totic mechanism has been demonstrated (Huang et al., Hum. disorder that occurs gradually and results in uncontrolled Mol. Genet. 6: 1879-1885, 1997). body movements, rigidity, tremor, and dyskinesia. These 0282. It is well-known that apoptosis plays a role in AIDS motor system problems are related to the death of brain cells pathogenesis in the immune system. However, HIV-1 also in an area of the brain that produces dopamine, a chemical that induces neurological disease. Shi et al. (J. Clin. Invest. 98: helps control muscle activity. In most people with PD, symp 1979-1990, 1996) examined apoptosis induced by HIV-1 toms appear after age 50. The initial symptoms of PD are a infection of the CNS in an in vitro model and in brain tissue pronounced tremor affecting the extremities, notably in the from AIDS patients, and found that HIV-1 infection of pri hands or lips. Subsequent characteristic symptoms of PD are mary brain cultures induced apoptosis in neurons and astro stiffness or slowness of movement, a shuffling walk, Stooped cytes in vitro. Apoptosis of neurons and astrocytes was also posture, and impaired balance. There are wide ranging sec detected in brain tissue from 10/11 AIDS patients, including ondary symptoms such as memory loss, dementia, depres 5/5 patients with HIV-1 dementia and 4/5 nondemented Sion, emotional changes, Swallowing difficulties, abnormal patients. speech, sexual dysfunction, and bladder and bowel problems. These symptoms will begin to interfere with routine activi 0283. There are four main peripheral neuropathies associ ties, such as holding a fork or reading a newspaper. Finally, ated with HIV, namely sensory neuropathy, AIDP/CIPD, people with PD become so profoundly disabled that they are drug-induced neuropathy and CMV-related. bedridden. 0284. The most common type of neuropathy associated 0278 ALS (motor neuron disease) is a chronic, incurable, with AIDS is distal symmetrical polyneuropathy (DSPN). and unstoppable CNS disorder that attacks the motor neurons, This syndrome is a result of nerve degeneration and is char components of the CNS that connect the brain to the skeletal acterized by numbness and a sensation of pins and needles. muscles. In ALS, the motor neurons deteriorate and eventu DSPN causes few serious abnormalities and mostly results in ally die, and though a person's brain normally remains fully numbness or tingling of the feet and slowed reflexes at the functioning and alert, the command to move never reaches the ankles. It generally occurs with more severe immunosuppres muscles. Most people who get ALS are between 40 and 70 sion and is steadily progressive. Treatment with tricyclic anti years old. The first motor neurons that weaken are those depressants relieves symptoms but does not affect the under controlling the arms or legs. Those with ALS may have lying nerve damage. trouble walking, they may drop things, fall, slur their speech, 0285 Aless frequent, but more severe type of neuropathy and laugh or cry uncontrollably. Eventually the muscles in the is known as acute or chronic inflammatory demyelinating limbs begin to atrophy from disuse. This muscle weakness polyneuropathy (AIDP/CIDP). In AIDP/CIDP there is dam will become debilitating and a person will need a wheelchair age to the fatty membrane covering the nerve impulses. This or become unable to function out of bed. kind of neuropathy involves inflammation and resembles the 0279. The causes of these neurological diseases have muscle deterioration often identified with long-term use of remained largely unknown. They are conventionally defined AZT. It can be the first manifestation of HIV infection, where as distinct diseases, yet clearly show extraordinary similari the patient may not complain of pain, but fails to respond to ties in basic processes and commonly demonstrate overlap standard reflex tests. This kind of neuropathy may be associ ping symptoms far greater than would be expected by chance ated with seroconversion, in which case it can sometimes alone. Current disease definitions fail to properly deal with resolve spontaneously. It can serve as a sign of HIV infection the issue of overlap and a new classification of the neurode and indicate that it might be time to considerantiviral therapy. generative disorders has been called for. AIDP/CIDP may be auto-immune in origin. 0280 HD is another neurodegenerative disease resulting 0286 Drug-induced, or toxic, neuropathies can be very from genetically programmed degeneration of neurons in painful. Antiviral drugs commonly cause peripheral neuropa certain areas of the brain. This degeneration causes uncon thy, as do other drugs e.g. Vincristine, dilantin (an anti-seizure trolled movements, loss of intellectual faculties, and emo medication), high-dose vitamins, isoniazid, and folic acid tional disturbance. HD is a familial disease, passed from antagonists. Peripheral neuropathy is often used in clinical parent to child through a dominant mutation in the wild-type trials for antivirals as a dose-limiting side effect, which means gene. Some early symptoms of HD are mood Swings, depres that more drugs should not be administered. Additionally, the Sion, irritability or trouble driving, learning new things, use of Such drugs can exacerbate otherwise minor neuropa remembering a fact, or making a decision. As the disease thies. Usually, these drug-induced neuropathies are reversible progresses, concentration on intellectual tasks becomes with the discontinuation of the drug. increasingly difficult and the patient may have difficulty feed 0287 CMV causes several neurological syndromes in ing himself or herself and Swallowing. AIDS, including encephalitis, myelitis, and polyradiculopa 0281 Tay-Sachs disease and Sandhoff disease are gly thy. colipid storage diseases caused by the lack of lysosomal 0288 Neuronal loss is also a salient feature of prion dis B-hexosaminidase (Gravel et al., in The Metabolic Basis of eases, such as Creutzfeldt-Jakob disease in human, BSE in Inherited Disease, eds. Scriver et al., McGraw-Hill, New cattle (mad cow disease), Scrapie Disease in sheep and goats, York, pp. 2839-2879, 1995). In both disorders, GM2 ganglio and feline spongiform encephalopathy (FSE) in cats. Sirtuin side and related glycolipids.Substrates for hexosaminidase modulating compounds that increase the level and/or activity accumulate in the nervous system and trigger acute neurode of a sirtuin protein may be useful for treating or preventing generation. In the most severe forms, the onset of symptoms neuronal loss due to these prior diseases. US 2009/0143376 A1 Jun. 4, 2009

0289. In another embodiment, a sirtuin-modulating com mally along nerve fiber pathways in the brain and spinal cord. pound that increases the level and/or activity of a sirtuin This results in disruption of normal electrical conductivity protein may be used to treat or prevent any disease or disorder within the axons, fatigue and disturbances of vision, strength, involving axonopathy. Distal axonopathy is a type of periph coordination, balance, sensation, and bladder and bowel eral neuropathy that results from Some metabolic or toxic function. derangement of peripheral nervous system (PNS) neurons. It 0294 As such, MS is now a common and well-known is the most common response of nerves to metabolic or toxic neurological disorder that is characterized by episodic disturbances, and as Such may be caused by metabolic dis patches of inflammation and demyelination which can occur eases such as diabetes, renal failure, deficiency syndromes anywhere in the CNS. However, almost always without any Such as malnutrition and alcoholism, or the effects of toxins or involvement of the peripheral nerves associated therewith. drugs. The most common cause of distal axonopathy is dia Demyelination produces a situation analogous to that result betes, and the most common distal axonopathy is diabetic ing from cracks or tears in an insulator Surrounding an elec neuropathy. The most distal portions of axons are usually the trical cord. That is, when the insulating sheath is disrupted, first to degenerate, and axonal atrophy advances slowly the circuit is “short circuited' and the electrical apparatus towards the nerve's cell body. If the noxious stimulus is associated therewith will function intermittently or nor at all. removed, regeneration is possible, though prognosis Such loss of myelin surrounding nerve fibers results in short decreases depending on the duration and severity of the circuits in nerves traversing the brain and the spinal cord that stimulus. Those with distal axonopathies usually present with thereby result in symptoms of MS. It is further found that such symmetrical glove-stocking sensori-motor disturbances. demyelination occurs in patches, as opposed to along the Deep tendon reflexes and autonomic nervous system (ANS) entire CNS. In addition, such demyelination may be intermit functions are also lost or diminished in affected areas. tent. Therefore, such plaques are disseminated in both time 0290 Diabetic neuropathies are neuropathic disorders and space. that are associated with diabetes mellitus. These conditions 0295. It is believed that the pathogenesis involves a local usually result from diabetic microvascular injury involving disruption of the blood brain barrier which causes a localized Small blood vessels that Supply nerves (vasa nervorum). Rela immune and inflammatory response, with consequent dam tively common conditions which may be associated with age to myelin and hence to neurons. diabetic neuropathy include third nerve palsy, mononeuropa 0296 Clinically, MS exists in both sexes and can occurat thy; mononeuritis multiplex; diabetic amyotrophy; a painful any age. However, its most common presentation is in the polyneuropathy; autonomic neuropathy; and thoracoabdomi relatively young adult, often with a single focal lesion such as nal neuropathy. Clinical manifestations of diabetic neuropa a damage of the optic nerve, an area of anesthesia (loss of thy include, for example, sensorimotor polyneuropathy Such sensation), or paraesthesia (localize loss offeeling), or mus as numbness, sensory loss, dysesthesia and nighttime pain; cular weakness. In addition, Vertigo, double vision, localized autonomic neuropathy Such as delayed gastric emptying or pain, incontinence, and pain in the arms and legs may occur gastroparesis; and cranial neuropathy Such as oculomotor upon flexing of the neck, as well as a large variety of less (3rd) neuropathies or Mononeuropathies of the thoracic or common symptoms. lumbar spinal nerves. 0297. An initial attack of MS is often transient, and it may 0291 Peripheral neuropathy is the medical term for dam be weeks, months, or years before a further attack occurs. age to nerves of the peripheral nervous system, which may be Some individuals may enjoy a stable, relatively event free caused either by diseases of the nerve or from the side-effects condition for a great number of years, while other less fortu of systemic illness. Peripheral neuropathies vary in their pre nate ones may experience a continual downhill course ending sentation and origin, and may affect the nerve or the neuro in complete paralysis. There is, most commonly, a series of muscular junction. Major causes of peripheral neuropathy remission and relapses, in which each relapse leaves a patient include seizures, nutritional deficiencies, and HIV, though Somewhat worse than before. Relapses may be triggered by diabetes is the most likely cause. Mechanical pressure from stressful events, viral infections or toxins. Therein, elevated staying in one position for too long, a tumor, intraneural body temperature, i.e., a fever, will make the condition worse, hemorrhage, exposing the body to extreme conditions such as or as a reduction oftemperature by, for example, a cold bath, radiation, cold temperatures, or toxic Substances can also may make the condition better. cause peripheral neuropathy. 0298. In yet another embodiment, a sirtuin-modulating 0292. In an exemplary embodiment, a sirtuin-modulating compound that increases the level and/or activity of a sirtuin compound that increases the level and/or activity of a sirtuin protein may be used to treat trauma to the nerves, including, protein may be used to treat or prevent multiple Sclerosis trauma due to disease, injury (including Surgical interven (MS), including relapsing MS and monosymptomatic MS, tion), or environmental trauma (e.g., neurotoxins, alcohol and other demyelinating conditions, such as, for example, ism, etc.). chromic inflammatory demyelinating polyneuropathy 0299 Sirtuin-modulating compounds that increase the (CIDP), or symptoms associated therewith. level and/or activity of a sirtuin protein may also be useful to 0293 MS is a chronic, often disabling disease of the cen prevent, treat, and alleviate symptoms of various PNS disor tral nervous system. Various and converging lines of evidence ders, such as the ones described below. The PNS is composed point to the possibility that the disease is caused by a distur of the nerves that lead to or branch off from the spinal cord bance in the immune function, although the cause of this and CNS. The peripheral nerves handle a diverse array of disturbance has not been established. This disturbance per functions in the body, including sensory, motor, and auto mits cells of the immune system to “attack’ myelin, the fat nomic functions. When an individual has a peripheral neur containing insulating sheath that Surrounds the nerve axons opathy, nerves of the PNS have been damaged. Nerve damage located in the central nervous system (“CNS). When myelin can arise from a number of causes, such as disease, physical is damaged, electrical pulses cannot travel quickly or nor injury, poisoning, or malnutrition. These agents may affect US 2009/0143376 A1 Jun. 4, 2009 26 either afferent or efferent nerves. Depending on the cause of eral nerve dysfunction). Mononeuritis multiplex refers to a damage, the nerve cell axon, its protective myelin sheath, or condition characterized by multiple isolated nerve injuries. both may be injured or destroyed. Mononeuropathies may result from a wide variety of causes, 0300. The term “peripheral neuropathy encompasses a including ischemia; traumatic injury; compression; connec wide range of disorders in which the nerves outside of the tive tissue diseases; cumulative trauma disorders; and other brain and spinal cord peripheral nerves—have been dam conditions; Neuralgia (intense or aching pain that occurs aged. Peripheral neuropathy may also be referred to as along the course or distribution of a peripheral or cranial peripheral neuritis, or if many nerves are involved, the terms nerve); Peripheral Nervous System Neoplasms (neoplasms polyneuropathy or polyneuritis may be used. which arise from peripheral nerve tissue). This includes neu 0301 Peripheral neuropathy is a widespread disorder, and rofibromas; Schwannomas; granular cell tumors; and malig there are many underlying causes. Some of these causes are nant peripheral nerve sheath tumors (see DeVita Jr et al., common, such as diabetes, and others are extremely rare, Cancer: Principles and Practice of Oncology, 5th ed., pp 1750 Such as acrylamide poisoning and certain inherited disorders. 1); and Nerve Compression Syndromes (mechanical com The most common worldwide cause of peripheral neuropathy pression of nerves or nerve roots from internal or external is leprosy. Leprosy is caused by the bacterium Mycobacte causes. These may result in a conduction block to nerve rium leprae, which attacks the peripheral nerves of affected impulses, due to, for example, myelin sheath dysfunction, or people. axonal loss. The nerve and nerve sheath injuries may be 0302 Leprosy is extremely rare in the United States, caused by ischemia; inflammation; or a direct mechanical where diabetes is the most commonly known cause of periph effect; Neuritis (a general term indicating inflammation of a eral neuropathy. It has been estimated that more than 17 peripheral or cranial nerve). Clinical manifestation may million people in the United States and Europe have diabetes include pain; paresthesias; paresis; or hyperesthesia; Poly related polyneuropathy. Many neuropathies are idiopathic; no neuropathies (diseases of multiple peripheral nerves). The known cause can be found. The most common of the inherited various forms are categorized by the type of nerve affected peripheral neuropathies in the United States is Charcot (e.g., sensory, motor, or autonomic), by the distribution of Marie-Tooth disease, which affects approximately 125,000 nerve injury (e.g., distal VS. proximal), by nerve component persons. primarily affected (e.g., demyelinating vs. axonal), by etiol 0303 Another of the better known peripheral neuropa ogy, or by pattern of inheritance. thies is Guillain-Barrésyndrome, which arises from compli 0305. In another embodiment, a sirtuin activating com cations associated with viral illnesses, such as cytomegalovi pound may be used to treat or prevent chemotherapeutic rus, Epstein-Barr virus, and human immunodeficiency virus induced neuropathy. The sirtuin modulating compounds may (HIV), or bacterial infection, including Campylobacter jejuni be administered prior to administration of the chemothera and Lyme disease. The worldwide incidence rate is approxi peutic agent, concurrently with administration of the chemo mately 1.7 cases per 100,000 people annually. Other well therapeutic drug, and/or after initiation of administration of known causes of peripheral neuropathies include chronic the chemotherapeutic drug. If the sirtuin activating com alcoholism, infection of the varicella-zoster virus, botulism, pound is administered after the initiation of administration of and poliomyelitis. Peripheral neuropathy may develop as a the chemotherapeutic drug, it is desirable that the sirtuin primary symptom, or it may be due to another disease. For activating compound be administered prior to, or at the first example, peripheral neuropathy is only one symptom of dis signs, of chemotherapeutic induced neuropathy. eases such as amyloid neuropathy, certain cancers, or inher 0306 Chemotherapy drugs can, damage any part of the ited neurologic disorders. Such diseases may affect the PNS nervous system. Encephalopathy and myelopathy are fortu and the CNS, as well as other body tissues. nately very rare. Damage to peripheral nerves is much more 0304. Other PNS diseases treatable with sirtuin-modulat common and can be a side effect of treatment experienced by ing compounds that increase the level and/or activity of a people with cancers, such as lymphoma. Most of the neur sirtuin protein include: Brachial Plexus Neuropathies (dis opathy affects sensory rather than motor nerves. Thus, the eases of the cervical and first thoracic roots, nerve trunks, common symptoms are tingling, numbness or a loss of bal cords, and peripheral nerve components of the brachial ance. The longest nerves in the body seem to be most sensitive plexus. Clinical manifestations include regional pain, pares hence the fact that most patients will report numbness or pins thesia; muscle weakness, and decreased sensation in the and needles in their hands and feet. upper extremity. These disorders may be associated with 0307 The chemotherapy drugs which are most commonly trauma, including birth injuries; thoracic outlet syndrome; associated with neuropathy, are the Vinca alkaloids (anti neoplasms, neuritis, radiotherapy; and other conditions. See cancer drugs originally derived from a member of the peri Adams et al., Principles of Neurology, 6th ed., pp 1351-2); winkle the Vinca plant genus) and a platinum-containing Diabetic Neuropathies (peripheral, autonomic, and cranial drug called Cisplatin. The Vinca alkaloids include the drugs nerve disorders that are associated with diabetes mellitus). vinblastine, Vincristine and vindesine. Many combination These conditions usually result from diabetic microvascular chemotherapy treatments for lymphoma for example CHOP injury involving Small blood vessels that Supply nerves (vasa and CVP contain Vincristine, which is the drug known to nervorum). Relatively common conditions which may be cause this problem most frequently. Indeed, it is the risk of associated with diabetic neuropathy include third nerve neuropathy that limits the dose of Vincristine that can be palsy, mononeuropathy; mononeuritis multiplex; diabetic administered. amyotrophy; a painful polyneuropathy; autonomic neuropa 0308 Studies that have been performed have shown that thy; and thoracoabdominal neuropathy (see Adams et al., most patients will lose some reflexes in their legs as a result of Principles of Neurology, 6th ed, p 1325); mononeuropathies treatment with Vincristine and many will experience some (disease or trauma involving a single peripheral nerve in degree oftingling (paresthesia) in their fingers and toes. The isolation, or out of proportion to evidence of diffuse periph neuropathy does not usually manifest itself right at the start of US 2009/0143376 A1 Jun. 4, 2009 27 the treatment but generally comes on over a period of a few nerve cells. The major symptoms of these diseases are similar, weeks. It is not essential to stop the drug at the first onset of although not identical, and usually affect people in midlife. symptoms, but if the neuropathy progresses this may be nec Given the similarities in Symptoms, the polyglutamine dis essary. It is very important that patients should report Such eases are hypothesized to progress via common cellular mechanisms. In recent years, Scientists have made great symptoms to their doctors, as the nerve damage is largely strides in unraveling what the mechanisms are. reversible if the drug is discontinued. Most doctors will often 0311. Above a certain threshold, the greater the number of reduce the dose of Vincristine or switch to another form of glutamine repeats in a protein, the earlier the onset of disease Vinca alkaloid such as vinblastine or vindesine if the symp and the more severe the symptoms. This suggests that abnor toms are mild. Occasionally, the nerves Supplying the bowel mally long glutamine tracts render their host protein toxic to are affected causing abdominal pain and constipation. nerve cells. 0309. In another embodiment, a sirtuin activating com 0312 To test this hypothesis, scientists have generated pound may be used to treat or prevent a polyglutamine dis genetically engineered mice expressing proteins with long ease. Huntington's Disease (HD) and Spinocerebellar ataxia polyglutamine tracts. Regardless of whether the mice express type 1 (SCA1) are just two examples of a class of genetic full-length proteins or only those portions of the proteins diseases caused by dynamic mutations involving the expan containing the polyglutamine tracts, they develop symptoms sion of triplet sequence repeats. In reference to this common of polyglutamine diseases. This suggests that a long poly mechanism, these disorders are called trinucleotide repeat glutamine tract by itself is damaging to cells and does not diseases. At least 14 Such diseases are known to affect human have to be part of a functional protein to cause its damage. beings. Nine of them, including SCA1 and Huntington's dis 0313 For example, it is thought that the symptoms of ease, have CAG as the repeated sequence (see Table 2 below). SCA1 are not directly caused by the loss of normal ataxin-1 Since CAG codes for an amino acid called glutamine, these function but rather by the interaction between ataxin-1 and nine trinucleotide repeat disorders are collectively known as another protein called LANP. LANP is needed for nerve cells polyglutamine diseases. to communicate with one another and thus for their survival. 0310 Although the genes involved in different poly When the mutant ataxin-1 protein accumulates inside nerve glutamine diseases have little in common, the disorders they cells, it “traps' the LANP protein, interfering with its normal cause follow a strikingly similar course. Each disease is char function. After a while, the absence of LANP function acterized by a progressive degeneration of a distinct group of appears to cause nerve cells to malfunction.

TABLE 2 Summary of Polyglutamine Diseases. Normal Disease Gene Chromosomal Pattern of repeat repeat Disease l8le location inheritance Protein length length Spinobulbar AR Xq13-21 X-linked androgen 9-36 38-62 muscular recessive receptor atrophy (AR) (Kennedy disease) Huntington's HD 4p16.3 autosomal huntingtin 6-35 36-121 disease dominant Dentatorubral- DRPLA 12p13.31 autosomal atrophin-1 6-35 49-88 pallidoluysian dominant atrophy (Haw River syndrome) Spinocerebellar SCA1 6p23 autosomal ataxin-1 6-44 39-82 ataxia type 1 dominant Spinocerebellar SCA2 12q24.1 autosomal ataxin-2 15-31 36-63 ataxia type 2 dominant Spinocerebellar SCA3 14q32.1 autosomal ataxin-3 12-40 SS-84 ataxia type 3 dominant (Machado Joseph disease) Spinocerebellar SCA6 19p13 autosomal C1- 4-18 21-33 ataxia type 6 dominant voltage dependent calcium channel Subunit Spinocerebellar SCA7 3p12-13 autosomal ataxin-7 4-35 37-306 ataxia type 7 dominant Spinocerebellar SCA17 6q27 autosomal TATA 25-42 45-63 ataxia type 17 dominant binding protein US 2009/0143376 A1 Jun. 4, 2009 28

0314. Many transcription factors have also been found in botic stroke. Typically, thrombotic strokes result from the neuronal inclusions in different diseases. It is possible that blockage of a blood vessel by accumulated deposits. these transcription factors interact with the polyglutamine 0318. In another embodiment, the ischemic condition may containing proteins and then become trapped in the neuronal result from a disorder that occurs in a part of the subject's inclusions. This in turn might keep the transcription factors body outside of the central nervous system, but yet still causes from turning genes on and off as needed by the cell. Another a reduction in blood flow to the central nervous system. These observation is hypoacetylation of histones in affected cells. disorders may include, but are not limited to a peripheral This has led to the hypothesis that Class I/II Histone Deacety vascular disorder, a venous thrombosis, a pulmonary embo lase (HDAC I/II) inhibitors, which are known to increase lus, arrhythmia (e.g. atrial fibrillation), a myocardial infarc histone acetylation, may be a novel therapy for polyglutamine tion, a transient ischemic attack, unstable angina, or sickle diseases (U.S. patent application Ser. No. 10/476,627; cell anemia. Moreover, the central nervous system ischemic "Method of treating neurodegenerative, psychiatric, and condition may occur as result of the Subject undergoing a other disorders with deacetylase inhibitors'). Surgical procedure. By way of example, the Subject may be 0315. In yet another embodiment, the invention provides a undergoing heart Surgery, lung Surgery, spinal Surgery, brain method for treating or preventing neuropathy related to Surgery, vascular Surgery, abdominal Surgery, or organ trans ischemic injuries or diseases, such as, for example, coronary plantation Surgery. The organ transplantation Surgery may heart disease (including congestive heart failure and myocar include heart, lung, pancreas, kidney or liver transplantation dial infarctions), stroke, emphysema, hemorrhagic shock, Surgery. Moreover, the central nervous system ischemic con peripheral vascular disease (upper and lower extremities) and dition may occur as a result of a trauma or injury to a part of transplant related injuries. the subject's body outside the central nervous system. By way 0316. In certain embodiments, the invention provides a of example, the trauma or injury may cause a degree of method to treat a central nervous system cell to prevent dam bleeding that significantly reduces the total volume of blood age in response to a decrease in blood flow to the cell. Typi in the subject's body. Because of this reduced total volume, cally the severity of damage that may be prevented will the amount of blood flow to the central nervous system is depend in large part on the degree of reduction in blood flow concomitantly reduced. By way of further example, the to the cell and the duration of the reduction. By way of trauma or injury may also result in the formation of a vaso example, the normal amount of perfusion to brain gray matter occlusion that restricts blood flow to the central nervous sys in humans is about 60 to 70 mL/100 g of brain tissue/min. tem Death of central nervous system cells typically occurs when 0319. Of course it is contemplated that the sirtuin activat the flow of blood falls below approximately 8-10 mL/100g of ing compounds may be employed to treat the central nervous brain tissue/min, while at slightly higher levels (i.e. 20-35 system ischemic condition irrespective of the cause of the mL/100g of brain tissue/min) the tissue remains alive but not condition. In one embodiment, the ischemic condition results able to function. In one embodiment, apoptotic or necrotic from a vaso-occlusion. The vaso-occlusion may be any type cell death may be prevented. In still a further embodiment, of occlusion, but is typically a cerebral thrombosis or an ischemic-mediated damage. Such as cytoxic edema or central embolism. In a further embodiment, the ischemic condition nervous system tissue anoxemia, may be prevented. In each may result from a hemorrhage. The hemorrhage may be any embodiment, the central nervous system cell may be a spinal type of hemorrhage, but is generally a cerebral hemorrhage or cell or a brain cell. a Subararachnoid hemorrhage. In still another embodiment, 0317. Another aspect encompasses administrating a sir the ischemic condition may result from the narrowing of a tuin activating compound to a subject to treat a central ner vessel. Generally speaking, the vessel may narrow as a result Vous system ischemic condition. A number of central nervous of a vasoconstriction Such as occurs during vasospasms, or system ischemic conditions may be treated by the sirtuin due to arteriosclerosis. In yet another embodiment, the activating compounds described herein. In one embodiment, ischemic condition results from an injury to the brain or spinal the ischemic condition is a stroke that results in any type of cord. ischemic central nervous system damage. Such as apoptotic or 0320 In yet another aspect, a sirtuin activating compound necrotic cell death, cytoxic edema or central nervous system may be administered to reduce infarct size of the ischemic tissue anoxia. The stroke may impact any area of the brain or core following a central nervous system ischemic condition. be caused by any etiology commonly known to result in the Moreover, a sirtuin activating compound may also be benefi occurrence of a stroke. In one alternative of this embodiment, cially administered to reduce the size of the ischemic penum the stroke is a brain stem stroke. Generally speaking, brain bra or transitional Zone following a central nervous system stem strokes strike the brain stem, which control involuntary ischemic condition. life-support functions such as breathing, blood pressure, and 0321. In one embodiment, a combination drug regimen heartbeat. In another alternative of this embodiment, the may include drugs or compounds for the treatment or preven stroke is a cerebellar stroke. Typically, cerebellar strokes tion of neurodegenerative disorders or secondary conditions impact the cerebellum area of the brain, which controls bal associated with these conditions. Thus, a combination drug ance and coordination. In still another embodiment, the regimen may include one or more sirtuin activators and one or stroke is an embolic stroke. In general terms, embolic strokes more anti-neurodegeneration agents. For example, one or may impact any region of the brain and typically result from more sirtuin-activating compounds can be combined with an the blockage of an artery by a vaso-occlusion. In yet another effective amount of one or more of L-DOPA; a dopamine alternative, the stroke may be a hemorrhagic stroke. Like agonist; an adenosine A receptor antagonist; a COMT ischemic strokes, hemorrhagic stroke may impact any region inhibitor; a MAO inhibitor; an N NOS inhibitor; a sodium of the brain, and typically result from a ruptured blood vessel channel antagonist; a selective N-methyl D-aspartate characterized by a hemorrhage (bleeding) within or Surround (NMDA) receptor antagonist; an AMPA/kainate receptor ing the brain. In a further embodiment, the stroke is a throm antagonist; a calcium channel antagonist; a GABA-A recep

US 2009/0143376 A1 Jun. 4, 2009

(phenylmethyl)-4-piperidinyl-1-pro-panone; 1-(6- aminophenyl)-4-methyl-7,8-m-ethylenedioxy-5H-2,3-ben acetylamino-benzobthien-2-yl)-3-1-(phenylmethyl)-4- Zodiazepine hydrochloride; and 2,3-dihydroxy-6-nitro-7-sul piperid-inyl-1-propanone; 1-(5-acetylamino-benzobthien famoylbenzo-fcquinoxaline. 2-yl)-3-1-(phenylmethyl-)-4-piperidinyl-1-propanone; 0330 Exemplary sodium channel antagonists include 6-hydroxy-3-2-1-(phenylmethyl)-4-piperidin-yl)ethyl-1, ajmaline, procainamide, flecainide and riluzole. 2-benzisoxazole; 5-methyl-3-2-1-(phenylmethyl)-4-pip 0331 Exemplary matrix-metalloprotease inhibitors eridinyl-ethyl-1,2-benzisoxazole; 6-methoxy-3-2-1 (phe include 4-4-(4-fluorophenoxy)benzenesulfon-ylaminotet nylmethyl)-4-piperidinylet-hyl-1,2-benzisoxazole; rahydropyran-4-carboxylic acid hydroxyamide: 5-Methyl-5- 6-acetamide-3-2-1-(phenylmethyl)-4-piperidinyl-ethyl (4-(4-fluorophenoxy)-phenoxy)-pyrimidine-2,4,6-trione; 1.2-benzisoxazole, 6-amino-3-2-1-(phenylmethyl)-4-pip 5-n-Butyl-5-(4-(4-fluorophenoxy)-phenoxy)-pyrimidine-2, eridinylethyl-1-1,2-benzisoxazole; 6-(4-morpholinyl)-3- 4,6-trione and prinomistat. 2-1-(phenylmethyl)-4-piperidin-yl)ethyl-1,2- 0332 Poly(ADPribose) polymerase (PARP) is an abun benzisoxazole; 5,7-dihydro-3-2-1-(phenylmethyl)-4- dant nuclear enzyme which is activated by DNA strand single piperidi-nylethyl-6H-pyrrolo 4.5-f-1,2-benzisoxazol-6- breaks to synthesize poly (ADP ribose) from NAD. Under One, 3-2-1-(phenylmethyl)-4-piperidinylethyl-1,2- normal conditions, PARP is involved in base excision repair benzisothiazole; 3-2-1-(phenylmethyl)-4-piperidinyl caused by oxidative stress via the activation and recruitment ethenyl-1,2-benzisoxazole; 6-phenylamino-3-2-1- of DNA repair enzymes in the nucleus. Thus, PARP plays a (phenylmethyl)-4-piperidinylethyl-1-2,-benzisoxaz-ole; role in cell necrosis and DNA repair. PARP also participates 6-(2-thiazolyl)-3-2-1-(phenylmethyl)-4-piperidinylethyl in regulating cytokine expression that mediates inflamma 1,2-benzis-oxazole: 6-(2-oxazolyl)-3-2-1-(phenylmethyl)- tion. Under conditions where DNA damage is excessive (such 4-piperidinylethyl-1,2-benzisoxazole; 6-pyrrolidinyl-3-2- as by acute excessive exposure to a pathological insult), 1-(phenylmethyl)-4-piperidinylethyl-1-2-benzisoxazole; PARP is over-activated, resulting in cell-based energetic fail 5,7-dihydro-5,5-dimethyl-3-2-1-(phenylmethyl)-4-pip ure characterized by NAD depletion and leading to ATP con eridinylethyl-6H-pyrrolo4.5-f-1,2-benzisoxazole-6-one: Sumption, cellular necrosis, tissue injury, and organ damage/ 6,8-dihydro-3-2-1-(phenylmethyl)-4-piperidinylethyl failure. PARP is thought to contribute to neurodegeneration 7H-pyrrolo5,4-g-1,2-benzisoxazole-7-one: 3-2-1-(phe by depleting nicotinamide adenine dinucleotide (NAD+) nylmethyl)-4-piperidinylethyl-5,6-8-trihydro-7H-isox which then reduces adenosine triphosphate (ATP; Cosi and aZolo 4.5-g-quinolin-7-one: 1-benzyl-4-((5,6-dimethoxy-1- Marien, Ann. N.Y. Acad. Sci., 890:227, 1999) contributing to indanon)-2-yl)methylpiperidine, 1-benzyl-4-((5.6- cell death which can be prevented by PARP inhibitors. Exem dimethoxy-1-indanon)-2-ylidenyl)methylpiperidine, plory PARP inhibitors can be found in Southan and Szabo, 1-benzyl-4-((5-methoxy-1-indanon)-2-yl)methylp-iperi Current Medicinal Chemistry, 10:321, 2003. dine, 1-benzyl-4-((5,6-diethoxy-1-indanon)-2-yl)methylpip 0333 Exemplary inhibitors of p38 MAP kinase and c-jun eridine, 1-benzyl-4-((5.6-methylenedioxy-1-indanon)-2-yl) N-terminal kinases include pyridyl imidazoles, such as PD methylpiperidine, 1-(m-nitrobenzyl)-4-((5,6-dimethoxy-1- 169316, isomeric PD 169316, SB 203580, SB 202190, SB indanon)-2-yl)methylpiperidine, 1-cyclohexymethyl-4-((5. 220026, and RWJ 67657. Others are described in U.S. Pat. 6-dimethoxy-1-indanon)-2-yl)methylpiperidine, 1-(m- No. 6.288,089, and incorporated by reference herein. florobenzyl)-4-((5,6-dimethoxy-1-indanon)-2-yl) 0334. In an exemplary embodiment, a combination methylpiperidine, 1-benzyl-4-((5,6-dimethoxy-1-indanon)- therapy for treating or preventing MS comprises a therapeu 2-yl)propylpiperidine, and 1-benzyl-4-((5-isopropoxy-6- tically effective amount of one or more sirtuin-modulating methoxy-1-indanon)-2-yl)methylpiperidine. compounds that increase the level and/or activity of a sirtuin protein and one or more of AvoneXOR (interferon beta-1a). 0326 Exemplary calcium channel antagonists include dil Tysabri R (natalizumab), or Fumaderm R) (BG-12/Oral Fuma tiazem, omega-conotoxin GVIA, methoxyverapamil, amlo rate). dipine, felodipine, lacidipine, and mibefradil. 0335. In another embodiment, a combination therapy for 0327 Exemplary GABA-A receptor modulators include treating or preventing diabetic neuropathy or conditions asso clomethiazole; IDDB. gaboxadol (4,5,6,7-tetrahydroisox ciated therewith comprises atherapeutically effective amount aZoloS.4-cpyridin-3-ol); ganaxolone (3C-hydroxy-3B-me of one or more sirtuin-modulating compounds that increase thyl-5C.-pregnan-20-one); fengabine (2-(butylimino)-(2- the level and/or activity of a sirtuin protein and one or more of chlorophenyl)methyl-4-chlorophenol): 2-(4- tricyclic antidepressants (TCAS) (including, for example, methoxyphenyl)-2,5,6,7,8,9-hexahydro-pyrazolo 4.3-c. imipramine, amytriptyline, desipramine and nortriptyline), cinnolin-3-one; 7-cyclobutyl-6-(2-methyl-2H-1,2,4-triazol serotonin reuptake inhibitors (SSRIs) (including, for 3-ylmethoxy)-3-phenyl-1,2,4-triazolo 4,3-bipyridazine; example, fluoxetine, paroxetine, Sertralene, and citalopram) (3-fluoro-4-methylphenyl)-N-(4-1-(2-methylphenyl)me and antiepileptic drugs (AEDs) (including, for example, thyl-benzimidazol-2-yl)methyl)-N-pentylcarboxamide: gabapentin, carbamazepine, and topimirate). and 3-(aminomethyl)-5-methylhexanoic acid. 0336. In another embodiment, the invention provides a 0328 Exemplary potassium channel openers include dia method for treating or preventing a polyglutamine disease Zoxide, flupirtine, pinacidil, levcromakalim, rilmakalim, using a combination comprising at least one sirtuin activating chromakalim, PCO-400 and SKP-450 (2-2"(1", 3"-diox compound and at least one HDAC I/II inhibitor. Examples of olone)-2-methyl-4-(2-oxo-1'-pyrrolidinyl)-6-nitro-2H-1- HDAC I/II inhibitors include hydroxamic acids, cyclic pep benzopyra-n). tides, benzamides, short-chain fatty acids, and depudecin. 0329 Exemplary AMPA/kainate receptor antagonists 0337 Examples of hydroxamic acids and hydroxamic include 6-cyano-7-nitroquinoxalin-2,3-di-one (CNOX): acid derivatives, but are not limited to, trichostatin A (TSA), 6-nitro-7-Sulphamoylbenzof duinoxaline-2,3-dione suberoylanilide hydroxamic acid (SAHA), oxamflatin, (NBQX); 6,7-dinitroquinoxaline-2,3-dione (DNQX): 1-(4- suberic bishydroxamic acid (SBHA), m-carboxy-cinnamic US 2009/0143376 A1 Jun. 4, 2009 32 acid bishydroxamic acid (CBHA), valproic acid and pyroxa cific enzyme that inactivates procoagulant components. Cal mide. TSA was isolated as an antifungiantibiotic (Tsuji etal cium ions are involved in many of the component reactions. (1976) J. Antibiot (Tokyo) 29:1-6) and found to be a potent Blood coagulation follows either the intrinsic pathway, where inhibitor of mammalian HDAC (Yoshida et al. (1990).J. Biol. all of the protein components are present in blood, or the Chem. 265:17174-17179). The finding that TSA-resistant extrinsic pathway, where the cell-membrane protein tissue cell lines have an altered HDAC evidences that this enzyme is factor plays a critical role. Clot formation occurs when an important target for TSA. Other hydroxamic acid-based fibrinogen is cleaved by thrombin to form fibrin. Blood clots HDAC inhibitors, SAHA, SBHA, and CBHA are synthetic are composed of activated platelets and fibrin. compounds that are able to inhibit HDAC at micromolar 0341 Further, the formation of blood clots does not only concentration or lower in vitro or in vivo. Glicket al. (1999) limit bleeding in case of an injury (hemostasis), but may lead Cancer Res. 59:43924399. These hydroxamic acid-based to serious organ damage and death in the context of athero HDAC inhibitors all possess an essential structural feature: a Sclerotic diseases by occlusion of an important artery or vein. polar hydroxamic terminal linked through a hydrophobic Thrombosis is thus blood clot formation at the wrong time methylene spacer (e.g. 6 carbon at length) to anotherpolar site and place. It involves a cascade of complicated and regulated which is attached to a terminal hydrophobic moiety (e.g., biochemical reactions between circulating blood proteins benzene ring). Compounds developed having Such essential (coagulation factors), blood cells (in particular platelets), and features also fall within the scope of the hydroxamic acids elements of an injured vessel wall. that may be used as HDAC inhibitors. 0342. Accordingly, the present invention provides antico 0338 Cyclic peptides used as HDAC inhibitors are mainly agulation and antithrombotic treatments aiming at inhibiting cyclic tetrapeptides. Examples of cyclic peptides include, but the formation of blood clots in order to prevent or treat blood are not limited to, trapoxin A, apicidin and depsipeptide. coagulation disorders, such as myocardial infarction, stroke, Trapoxin A is a cyclic tetrapeptide that contains a 2-amino loss of a limb by peripheral artery disease or pulmonary 8-oxo-9,10-epoxy-decanoyl (AOE) moiety. Kijima et al. embolism. (1993).J. Biol. Chem. 268:22429-22435. Apicidin is a fungal 0343 As used interchangeably herein, "modulating or metabolite that exhibits potent, broad-spectrum antiprotozoal modulation of hemostasis' and “regulating or regulation of activity and inhibits HDAC activity at nanomolar concentra hemostasis includes the induction (e.g., stimulation or tions. Darkin-Rattray et al. (1996) Proc. Natl. Acad. Sci. increase) of hemostasis, as well as the inhibition (e.g., reduc USA. 93: 13143-13147. Depsipeptide is isolated from Chro tion or decrease) of hemostasis. mobacterium violaceum, and has been shown to inhibit 0344. In one aspect, the invention provides a method for HDAC activity at micromolar concentrations. reducing or inhibiting hemostasis in a Subject by administer 0339 Examples of benzamides include but are not limited ing a sirtuin-modulating compound that increases the level to MS-27-275. Saito et al. (1990) Proc. Natl. Acad. Sci. USA. and/or activity of a sirtuin protein. The compositions and 96:4592-4597. Examples of short-chain fatty acids include methods disclosed herein are useful for the treatment or pre but are not limited to butyrates (e.g., butyric acid, arginine vention of thrombotic disorders. As used herein, the term butyrate and phenylbutyrate (PB)). Newmark et al. (1994) “thrombotic disorder includes any disorder or condition Cancer Lett. 78:1-5; and Carducci et al. (1997) Anticancer characterized by excessive or unwanted coagulation or hemo Res. 17:3972-3973. In addition, depudecin which has been static activity, or a hypercoagulable state. Thrombotic disor shown to inhibit HDAC at micromolar concentrations (Kwon ders include diseases or disorders involving platelet adhesion et al. (1998) Proc. Natl. Acad. Sci. USA.95:3356-3361) also and thrombus formation, and may manifest as an increased falls within the scope of histone deacetylase inhibitor as propensity to form thromboses, e.g., an increased number of described herein. thromboses, thrombosis at an early age, a familial tendency towards thrombosis, and thrombosis at unusual sites. Blood Coagulation Disorders Examples of thrombotic disorders include, but are not limited 0340. In other aspects, sirtuin-modulating compounds that to, thromboembolism, deep vein thrombosis, pulmonary increase the level and/or activity of a sirtuin protein can be embolism, stroke, myocardial infarction, miscarriage, throm used to treat or prevent blood coagulation disorders (or hemo bophilia associated with anti-thrombin III deficiency, protein static disorders). As used interchangeably herein, the terms C deficiency, protein S deficiency, resistance to activated "hemostasis”, “blood coagulation, and “blood clotting refer protein C, dysfibrinogenemia, fibrinolytic disorders, to the control of bleeding, including the physiological prop homocystinuria, pregnancy, inflammatory disorders, myelo erties of vasoconstriction and coagulation. Blood coagulation proliferative disorders, arteriosclerosis, angina, e.g., unstable assists in maintaining the integrity of mammalian circulation angina, disseminated intravascular coagulation, thrombotic after injury, inflammation, disease, congenital defect, dys thrombocytopenic purpura, cancer metastasis, sickle cell dis function or other disruption. After initiation of clotting, blood ease, glomerular nephritis, and drug induced thrombocytope coagulation proceeds through the sequential activation of nia (including, for example, heparin induced thrombocytope certain plasma proenzymes to their enzyme forms (see, for nia). In addition, sirtuin-modulating compounds that increase example, Coleman, R. W. et al. (eds.) Hemostasis and Throm the level and/or activity of a sirtuin protein may be adminis bosis, Second Edition, (1987)). These plasma glycoproteins, tered to prevent thrombotic events or to prevent re-occlusion including Factor XII, Factor XI, Factor IX, Factor X, Factor during or after therapeutic clot lysis or procedures Such as VII, and prothrombin, are Zymogens of serine proteases. angioplasty or Surgery. Most of these blood clotting enzymes are effective on a physi 0345. In another embodiment, a combination drug regi ological scale only when assembled in complexes on mem men may include drugs or compounds for the treatment or brane surfaces with protein cofactors such as Factor VIII and prevention of blood coagulation disorders or secondary con Factor V. Other blood factors modulate and localize clot for ditions associated with these conditions. Thus, a combination mation, or dissolve blood clots. Activated protein C is a spe drug regimen may include one or more sirtuin-modulating US 2009/0143376 A1 Jun. 4, 2009 compounds that increase the level and/or activity of a sirtuin protein may be used for reducing appetite and/or increasing protein and one or more anti-coagulation or anti-thrombosis Satiety, thereby causing weight loss or avoidance of weight agents. For example, one or more sirtuin-modulating com gain. A subject in need of Such a treatment may be a subject pounds can be combined with an effective amount of one or who is overweight, obese or a subject likely to become over more of aspirin, heparin, and oral Warfarin that inhibits Vit weight or obese. The method may comprise administering K-dependent factors, low molecular weight heparins that daily or, every other day, or once a week, a dose, e.g., in the inhibit factors X and II, thrombin inhibitors, inhibitors of form of a pill, to a subject. The dose may be an “appetite platelet GP IIb IIIa receptors, inhibitors of tissue factor (TF), reducing dose.” inhibitors of human von Willebrand factor, inhibitors of one 0350. In other embodiments, a sirtuin-modulating com or more factors involved in hemostasis (in particular in the pound that decreases the level and/or activity of a sirtuin coagulation cascade). In addition, sirtuin-modulating com protein may be used to stimulate appetite and/or weight gain. pounds that increase the level and/or activity of a sirtuin A method may comprise administering to a subject, Such as a protein can be combined with thrombolytic agents, such as Subject in need thereof, a pharmaceutically effective amount t-PA, streptokinase, reptilase, TNK-t-PA, and staphyloki of a sirtuin-modulating agent that decreases the level and/or aSC. activity of a sirtuin protein, such as SIRT1 and/or SIRT3. A Subject in need of Such a treatment may be a subject who has Weight Control cachexia or may be likely to develop cachexia. A combination 0346. In another aspect, sirtuin-modulating compounds of agents may also be administered. A method may further that increase the level and/or activity of a sirtuin protein may comprise monitoring in the Subject the state of the disease or be used for treating or preventing weight gain or obesity in a of activation of Sirtuins, for example, in adipose tissue. Subject. For example, sirtuin-modulating compounds that 0351 Methods for stimulating fat accumulation in cells increase the level and/or activity of a sirtuin protein may be may be used in vitro, to establish cell models of weight gain, used, for example, to treat or prevent hereditary obesity, which may be used, e.g., for identifying other drugs that dietary obesity, hormone related obesity, obesity related to prevent weight gain. the administration of medication, to reduce the weight of a 0352 Also provided are methods for modulating adipo Subject, or to reduce or prevent weight gain in a Subject. A genesis or fat cell differentiation, whether in vitro or in vivo. Subject in need of Such a treatment may be a subject who is In particular, high circulating levels of insulin and/or insulin obese, likely to become obese, overweight, or likely to like growth factor (IGF) 1 will be prevented from recruiting become overweight. Subjects who are likely to become obese preadipocytes to differentiate into adipocytes. Such methods or overweight can be identified, for example, based on family may be used to modulate obesity. A method for stimulating history, genetics, diet, activity level, medication intake, or adipogenesis may comprise contacting a cell with a sirtuin various combinations thereof. modulating agent that decreases the level and/or activity of a 0347 In yet other embodiments, sirtuin-modulating com sirtuin protein. pounds that increase the level and/or activity of a sirtuin 0353. In another embodiment, the invention provides protein may be administered to Subjects suffering from a methods of decreasing fat or lipid metabolism in a subject by variety of other diseases and conditions that may be treated or administering a sirtuin-modulating compound that decreases prevented by promoting weight loss in the Subject. Such the level and/or activity of a sirtuin protein. The method diseases include, for example, high blood pressure, hyperten includes administering to a subject an amount of a sirtuin Sion, high blood cholesterol, dyslipidemia, type 2 diabetes, modulating compound, e.g., in an amount effective to insulin resistance, glucose intolerance, hyperinsulinemia, decrease mobilization of fat to the blood from WAT cells coronary heart disease, angina pectoris, congestive heart fail and/or to decrease fat burning by BAT cells. ure, stroke, gallstones, cholescystitis and cholelithiasis, gout, 0354 Methods for promoting appetite and/or weight gain osteoarthritis, obstructive sleep apnea and respiratory prob may include, for example, prior identifying a subject as being lems. Some types of cancer (such as endometrial, breast, in need of decreased fat or lipid metabolism, e.g., by weighing prostate, and colon), complications of pregnancy, poor the subject, determining the BMI of the subject, or evaluating female reproductive health (such as menstrual irregularities, fat content of the subject or sirtuin activity in cells of the infertility, irregular ovulation), bladder control problems Subject. The method may also include monitoring the Subject, (such as stress incontinence); uric acid nephrolithiasis; psy e.g., during and/or after administration of a sirtuin-modulat chological disorders (such as depression, eating disorders, ing compound. The administering can include one or more distorted body image, and low self esteem). Stunkard A. J. dosages, e.g., delivered in boluses or continuously. Monitor Wadden T.A. (Editors) Obesity: theory and therapy, Second ing can include evaluating a hormone or a metabolite. Exem Edition. New York: Raven Press, 1993. Finally, patients with plary hormones include leptin, adiponectin, resistin, and AIDS can develop lipodystrophy or insulin resistance in insulin. Exemplary metabolites include triglyercides, choles response to combination therapies for AIDS. terol, and fatty acids. 0348. In another embodiment, sirtuin-modulating com 0355. In one embodiment, a sirtuin-modulating com pounds that increase the level and/or activity of a sirtuin pound that decreases the level and/or activity of a sirtuin protein may be used for inhibiting adipogenesis or fat cell protein may be used to modulate (e.g., increase) the amount differentiation, whether in vitro or in vivo. In particular, high of subcutaneous fat in a tissue, e.g., in facial tissue or in other circulating levels of insulin and/or insulin like growth factor Surface-associated tissue of the neck, hand, leg, or lips. The (IGF) 1 will be prevented from recruiting preadipocytes to sirtuin-modulating compound may be used to increase the differentiate into adipocytes. Such methods may be used for rigidity, water retention, or Support properties of the tissue. treating or preventing obesity. For example, the sirtuin-modulating compound can be 0349. In other embodiments, sirtuin-modulating com applied topically, e.g., in association with another agent, e.g., pounds that increase the level and/or activity of a sirtuin for Surface-associated tissue treatment. The sirtuin-modulat US 2009/0143376 A1 Jun. 4, 2009 34 ing compound may also be injected Subcutaneously, e.g., as insulin-resistance, a pre-diabetic state, type II diabetes, within the region where an alteration in Subcutaneous fat is and/or complications thereof. Administration of a sirtuin desired. modulating compounds that increases the level and/or activ 0356. A method for modulating weight may further com ity of a sirtuin protein may increase insulin sensitivity and/or prise monitoring the weight of the subject and/or the level of decrease insulin levels in a subject. A Subject in need of Such modulation of sirtuins, for example, in adipose tissue. a treatment may be a Subject who has insulin resistance or 0357. In an exemplary embodiment, sirtuin-modulating other precursor symptom of type II diabetes, who has type II compounds that increase the level and/or activity of a sirtuin diabetes, or who is likely to develop any of these conditions. protein may be administered as a combination therapy for For example, the Subject may be a subject having insulin treating or preventing weight gain or obesity. For example, resistance, e.g., having high circulating levels of insulin and/ one or more sirtuin-modulating compounds that increase the or associated conditions, such as hyperlipidemia, dyslipogen level and/or activity of a sirtuin protein may be administered esis, hypercholesterolemia, impaired glucose tolerance, high in combination with one or more anti-obesity agents. Exem blood glucose Sugar level, other manifestations of syndrome plary anti-obesity agents include, for example, phenylpro X, hypertension, atherosclerosis and lipodystrophy. panolamine, ephedrine, pseudoephedrine, phentermine, a 0360. In an exemplary embodiment, sirtuin-modulating cholecystokinin-A agonist, a monoamine reuptake inhibitor compounds that increase the level and/or activity of a sirtuin (such as Sibutramine), a sympathomimetic agent, a seroton protein may be administered as a combination therapy for ergic agent (Such as dexfenfluramine or fenfluramine), a treating or preventing a metabolic disorder. For example, one dopamine agonist (such as bromocriptine), a melanocyte or more sirtuin-modulating compounds that increase the level stimulating hormone receptor agonist or mimetic, a melano and/or activity of a sirtuin protein may be administered in cyte-stimulating hormone analog, a cannabinoid receptor combination with one or more anti-diabetic agents. Exem antagonist, a melanin concentrating hormone antagonist, the plary anti-diabetic agents include, for example, an aldose OB protein (leptin), a leptin analog, a leptin receptoragonist, reductase inhibitor, a glycogen phosphorylase inhibitor, a agalaninantagonist or a GIlipase inhibitor or decreaser (Such Sorbitol dehydrogenase inhibitor, a protein tyrosine phos as orlistat). Other anorectic agents include bombesin ago phatase 1B inhibitor, a dipeptidyl protease inhibitor, insulin nists, dehydroepiandrosterone or analogs thereof glucocor (including orally bioavailable insulin preparations), an insu ticoid receptor agonists and antagonists, orexin receptor lin mimetic, metformin, acarbose, a peroxisome proliferator antagonists, urocortin binding protein antagonists, agonists activated receptor-Y (PPAR-Y) ligand such as troglitazone, of the glucagon-like peptide-1 receptor Such as Exendin and rosaglitazone, pioglitazone or GW-1929, a sulfonylurea, gli ciliary neurotrophic factors such as AXokine. pazide, glyburide, or chlorpropamide wherein the amounts of 0358. In another embodiment, sirtuin-modulating com the first and second compounds result in a therapeutic effect. pounds that increase the level and/or activity of a sirtuin Other anti-diabetic agents include a glucosidase inhibitor, a protein may be administered to reduce drug-induced weight glucagon-like peptide-1 (GLP-1), insulin, a PPAR C/y dual gain. For example, a sirtuin-modulating compound that agonist, a meglitimide and an OP2 inhibitor. In an exemplary increases the level and/or activity of a sirtuin protein may be embodiment, an anti-diabetic agent may be a dipeptidyl pep administered as a combination therapy with medications that tidase IV (DP-UV or DPP-IV) inhibitor, such as, for example may stimulate appetite or cause weight gain, in particular, LAF237 from Novartis (NVP DPP728; 1-2-(5-cyanopy weight gain due to factors other than water retention. ridin-2-yl)aminoethylaminoacetyl-2-cyano-(S)-pyrroli Examples of medications that may cause weight gain, include dine) or MK-04301 from Merck (see e.g., Hughes et al., for example, diabetes treatments, including, for example, Sul Biochemistry 38: 11597-603 (1999)). fonylureas (such as glipizide and glyburide), thiazolidinedi ones (such as pioglitaZone and rosiglitaZone), meglitinides, Inflammatory Diseases nateglinide, repaglinide, Sulphonylurea medicines, and insu 0361. In other aspects, sirtuin-modulating compounds that lin; anti-depressants, including, for example, tricyclic antide increase the level and/or activity of a sirtuin protein can be pressants (such as amitriptyline and imipramine), irreversible used to treat or prevent a disease or disorder associated with monoamine oxidase inhibitors (MAOIs), selective serotonin inflammation. Sirtuin-modulating compounds that increase reuptake inhibitors (SSRIs), bupropion, paroxetine, and mir the level and/or activity of a sirtuin protein may be adminis tazapine; Steroids. Such as, for example, prednisone; hormone tered prior to the onset of at, or after the initiation of inflam therapy; lithium carbonate; Valproic acid; carbamazepine; mation. When used prophylactically, the compounds are pref chlorpromazine; thiothixene; beta blockers (such as propra erably provided in advance of any inflammatory response or nolo); alpha blockers (such as clonidine, praZosin and tera symptom. Administration of the compounds may prevent or Zosin); and contraceptives including oral contraceptives attenuate inflammatory responses or symptoms. (birth control pills) or other contraceptives containing estro 0362 Exemplary inflammatory conditions include, for gen and/or progesterone (Depo-Provera, Norplant, Ortho), example, multiple Sclerosis, rheumatoid arthritis, psoriatic testosterone or Megestrol. In another exemplary embodi arthritis, degenerative joint disease, spondouloarthropathies, ment, sirtuin-modulating compounds that increase the level gouty arthritis, Systemic lupus erythematosus, juvenile arthri and/or activity of a sirtuin protein may be administered as part tis, rheumatoid arthritis, osteoarthritis, osteoporosis, diabetes of a Smoking cessation program to prevent weight gain or (e.g., insulin dependent diabetes mellitus or juvenile onset reduce weight already gained. diabetes), menstrual cramps, cystic fibrosis, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, Metabolic Disorders/Diabetes mucous colitis, ulcerative colitis, gastritis, esophagitis, pan 0359. In another aspect, sirtuin-modulating compounds creatitis, peritonitis, Alzheimer's disease, shock, ankylosing that increase the level and/or activity of a sirtuin protein may spondylitis, gastritis, conjunctivitis, pancreatis (acute or be used for treating or preventing a metabolic disorder. Such chronic), multiple organ injury syndrome (e.g., secondary to US 2009/0143376 A1 Jun. 4, 2009 septicemia or trauma), myocardial infarction, atherosclero late, benoxaprofen, benzpiperylon, benzydamine, benzyl sis, stroke, reperfusion injury (e.g., due to cardiopulmonary morphine, bermoprofen, betamethasone, betamethasone-17 bypass or kidney dialysis), acute glomerulonephritis, Vascu Valerate, bezitramide, C.-bisabolol, bromfenac, litis, thermal injury (i.e., Sunburn), necrotizing enterocolitis, p-bromoacetanilide, 5-bromosalicylic acid acetate, bromo granulocyte transfusion associated syndrome, and/or saligenin, bucetin, bucloxic acid, bucolome, budesonide, Sjogren's syndrome. Exemplary inflammatory conditions of bufexamac, bumadizon, buprenorphine, butacetin, butibufen, the skin include, for example, eczema, atopic dermatitis, con butorphanol, carbamazepine, carbiphene, carprofen, tact dermatitis, urticaria, Schleroderma, psoriasis, and derma carsalam, chlorobutanol, chloroprednisone, chlorthenoxazin, tosis with acute inflammatory components. choline salicylate, cinchophen, cinmetacin, ciramadol, clid 0363. In another embodiment, sirtuin-modulating com anac, clobetasol, clocortolone, clometacin, clonitaZene, pounds that increase the level and/or activity of a sirtuin clonixin, clopirac, cloprednol, clove, codeine, codeine protein may be used to treat or prevent allergies and respira methyl bromide, codeine phosphate, codeine Sulfate, corti tory conditions, including asthma, bronchitis, pulmonary Sone, cortivaZol, cropropamide, crotethamide, cyclazocine, fibrosis, allergic rhinitis, oxygen toxicity, emphysema, deflazacort, dehydrotestosterone, desomorphine, desonide, chronic bronchitis, acute respiratory distress syndrome, and desoximetaSone, dexamethasone, dexamethasone-21-isoni any chronic obstructive pulmonary disease (COPD). The cotinate, deXOXadrol, dextromoramide, dextropropoxyphene, compounds may be used to treat chronic hepatitis infection, deoxycorticosterone, dezocine, diampromide, diamorphone, including hepatitis B and hepatitis C. diclofenac, difenamizole, difenpiramide, diflorasone, diflu 0364. Additionally, sirtuin-modulating compounds that cortolone, diflunisal, difluprednate, dihydrocodeine, dihy increase the level and/or activity of a sirtuin protein may be drocodeinone enol acetate, dihydromorphine, dihydroxyalu used to treat autoimmune diseases and/or inflammation asso minum acetylsalicylate, dimenoxadol, dimepheptanol, ciated with autoimmune diseases such as organ-tissue dimethylthiambutene, dioxaphetyl butyrate, dipipanone, autoimmune diseases (e.g., Raynaud's syndrome), Sclero diprocetyl, dipyrone, ditazol, droxicam, emorfaZone, enfe derma, myasthenia gravis, transplant rejection, endotoxin namic acid, enoXolone, epirizole, eptazocine, etersalate, shock, sepsis, psoriasis, eczema, dermatitis, multiple Sclero ethenZamide, ethoheptazine, ethoxazene, ethylmethylthiam sis, autoimmune thyroiditis, uveitis, systemic lupus erythe butene, ethylmorphine, etodolac, etofenamate, etonitaZene, matosis, Addison's disease, autoimmune polyglandular dis eugenol, felbinac, fenbufen, fenclozic acid, fendosal, feno ease (also known as autoimmune polyglandular syndrome), profen, fentanyl, fentiazac, fepradinol, feprazone, floctafe and Grave's disease. nine, fluazacort, flucloronide, flufenamic acid, flumethasone, 0365. In certain embodiments, one or more sirtuin-modu flunisolide, flunixin, flunoxaprofen, fluocinolone acetonide, lating compounds that increase the level and/or activity of a fluocinonide, fluocinolone acetonide, fluocortin butyl, fluo sirtuin protein may be taken alone or in combination with cortolone, fluoresone, fluorometholone, fluperolone, flupir other compounds useful for treating or preventing inflamma tine, fluprednidene, fluprednisolone, fluproduaZone, fluran tion. Exemplary anti-inflammatory agents include, for drenolide, flurbiprofen, fluticasone, formocortal, fosfosal, example, steroids (e.g., cortisol, cortisone, fludrocortisone, gentisic acid, glafenine, glucametacin, glycol Salicylate, gua prednisone, 6.C.-methylprednisone, triamcinolone, iaZulene, halcinonide, halobetasol, halometaSone, halopred betamethasone or dexamethasone), nonsteroidal antiinflam none, heroin, hydrocodone, hydrocortamate, hydrocortisone, matory drugs (NSAIDS (e.g., aspirin, acetaminophen, tol hydrocortisone acetate, hydrocortisone Succinate, hydrocor metin, ibuprofen, mefenamic acid, piroxicam, nabumetone, tisone hemisuccinate, hydrocortisone 21-lysinate, hydrocor rofecoxib, celecoxib, etodolac or nimeSulide). In another tisone cypionate, hydromorphone, hydroxypethidine, embodiment, the other therapeutic agent is an antibiotic (e.g., ibufenac, ibuprofen, ibuproxam, imidazole salicylate, Vancomycin, penicillin, amoxicillin, amplicillin, cefotaxime, indomethacin, indoprofen, isofeZolac, isoflupredone, isoflu ceftriaxone, cefixime, rifampinmetronidazole, doxycycline predone acetate, isoladol, isomethadone, isonixin, isoxepac, or streptomycin). In another embodiment, the other therapeu isoxicam, ketobemidone, ketoprofen, ketorolac, p-lactophe tic agent is a PDE4 inhibitor (e.g., roflumilast or rolipram). In netide, lefetamine, levallorphan, levorphanol, levophenacyl another embodiment, the other therapeutic agent is an anti morphan, lofentanil, lonazolac, lornoxicam, loxoprofen, histamine (e.g., cyclizine, hydroxy Zine, promethazine or lysine acetylsalicylate, maZipredone, meclofenamic acid, diphenhydramine). In another embodiment, the other thera medrysone, mefenamic acid, meloxicam, meperidine, peutic agent is an anti-malarial (e.g., artemisinin, artemether, meprednisone, meptazinol, mesalamine, metazocine, metha artsunate, chloroquine phosphate, mefloquine hydrochloride, done, methotrimeprazine, methylprednisolone, methylpred doxycycline hyclate, proguanil hydrochloride, atovaquone or nisolone acetate, methylprednisolone sodium Succinate, halofantrine). In one embodiment, the other therapeutic agent methylprednisolone Suleptanate, metiazinic acid, metofoline, is drotrecogin alfa. metopon, mofebutaZone, mofeZolac, mometasone, mora 0366 Further examples of anti-inflammatory agents Zone, morphine, morphine hydrochloride, morphine Sulfate, include, for example, aceclofenac, acemetacin, e-acetami morpholine Salicylate, myrophine, nabumetone, nalbuphine, docaproic acid, acetaminophen, acetaminosalol, acetanilide, nalorphine, 1-naphthyl salicylate, naproxen, narceline, nefo acetylsalicylic acid, S-adenosylmethionine, alclofenac, pam, nicomorphine, nifenaZone, niflumic acid, nimeSulide, alclometasone, alfentanil, algestone, allylprodine, almino 5'-nitro-2'-propoxyacetanilide, norlevorphanol, normetha profen, aloxiprin, alphaprodine, aluminum bis(acetylsalicy done, normorphine, norpipanone, olsalazine, opium, late), amcinonide, amfenac, aminochlorthenoxazin, oxaceprol, oxametacine, oxaprozin, oxycodone, oxymor 3-amino-4-hydroxybutyric acid, 2-amino-4-picoline, amino phone, oxyphenbutaZone, papaveretum, paramethasone, propylon, aminopyrine, amiXetrine, ammonium salicylate, paranyline, parsalmide, pentazocine, perisoxal, phenacetin, ampiroXicam, amtolimetin guacil, anilleridine, antipyrine, phenadoxone, phenazocine, phenazopyridine hydrochloride, antrafenine, apaZone, beclomethasone, bendazac, benory phenocoll, phenoperidine, phenopyrazone, phenomorphan, US 2009/0143376 A1 Jun. 4, 2009 36 phenyl acetylsalicylate, phenylbutaZone, phenyl salicylate, modulating compound may be administered (1) at the same as phenyramidol, piketoprofen, piminodine, pipebuZone, pip administration of the flushing inducing agent, (2) intermit erylone, piraZolac, piritramide, piroXicam, pirprofen, prano tently with the flushing inducing agent, (3) staggered relative profen, prednicarbate, prednisolone, prednisone, prednival, to administration of the flushing inducing agent, (4) prior to prednylidene, proglumetacin, proheptazine, promedol, pro administration of the flushing inducing agent, (5) Subsequent pacetamol, properidine, propiram, propoxyphene, propy to administration of the flushing inducing agent, and (6) vari phenaZone, produaZone, protizinic acid, proxazole, ram ous combination thereof. Exemplary flushing inducing ifenaZone, remifentanil, rimazolium metilsulfate, agents include, for example, niacin, faloxifene, antidepres salacetamide, Salicin, Salicylamide, salicylamide o-acetic sants, anti-psychotics, chemotherapeutics, calcium channel acid, salicylic acid, salicylsulfuric acid, Salsalate, salverine, simetride, Sufentanil, Sulfasalazine, Sulindac, Superoxide dis blockers, and antibiotics. mutase, Suprofen, SuXibuZone, talniflumate, tenidap, tenoxi 0370. In one embodiment, sirtuin-modulating compounds cam, terofenamate, tetrandrine, thiazolinobutaZone, tiapro that increase the level and/or activity of a sirtuin protein may fenic acid, tiaramide, tilidine, tinoridine, tiXocortol, be used to reduce flushing side effects of a vasodilator or an tolfenamic acid, tolmetin, tramadol, triamcinolone, triamci antilipidemic agent (including anticholesteremic agents and nolone acetonide, tropesin, Viminol, Xenbucin, Ximoprofen, lipotropic agents). In an exemplary embodiment, a sirtuin Zaltoprofen and Zomepirac. modulating compound that increases the level and/or activity 0367. In an exemplary embodiment, a sirtuin-modulating of a sirtuin protein may be used to reduce flushing associated compound that increases the level and/or activity of a sirtuin with the administration of niacin. protein may be administered with a selective COX-2 inhibitor 0371 Nicotinic acid, 3-pyridinecarboxylic acid or niacin, for treating or preventing inflammation. Exemplary selective is an antilipidemic agent that is marketed under, for example, COX-2 inhibitors include, for example, deracoxib, pare the trade names Nicolar R, SloNiacin R, NicobidR and Time coxib, celecoxib, Valdecoxib, rofecoxib, etoricoxib, lumira Release Niacinr. Nicotinic acid has been used for many coxib, 2-(3,5-difluorophenyl)-3-4-(methylsulfonyl)phe years in the treatment of lipidemic disorders such as hyper nyl-2-cyclopenten-1-one, (S)-6,8-dichloro-2-(triflu lipidemia, hypercholesterolemia and atherosclerosis. This oromethyl)-2H-1-benzopyran-3-carboxylic acid, 2-(3,4- compound has long been known to exhibit the beneficial difluorophenyl)-4-(3-hydroxy-3-methyl-1-butoxy)-5-4- effects of reducing total cholesterol, low density lipoproteins (methylsulfonyl)phenyl)-3-(2H)-pyridazinone, 4-5-(4- or “LDL cholesterol.” triglycerides and apolipoproteina (Lp fluorophenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl) (a)) in the human body, while increasing desirable high den benzenesulfonamide, tert-butyl 1 benzyl-4-(4-oxopiperidin sity lipoproteins or “HDL cholesterol. 1-yl)sulfonylpiperidine-4-carboxylate, 4-5-(phenyl)-3- 0372 Typical doses range from about 1 gram to about 3 (trifluoromethyl)-1H-pyrazol-1-ylbenzenesulfonamide, grams daily. Nicotinic acid is normally administered two to salts and prodrugs thereof. four times per day after meals, depending upon the dosage form selected. Nicotinic acid is currently commercially avail Flushing able in two dosage forms. One dosage form is an immediate 0368. In another aspect, sirtuin-modulating compounds or rapid release tablet which should be administered three or that increase the level and/or activity of a sirtuin protein may four times per day. Immediate release (“IR”) nicotinic acid be used for reducing the incidence or severity of flushing formulations generally release nearly all of their nicotinic and/or hot flashes which are symptoms of a disorder. For acid within about 30 to 60 minutes following ingestion. The instance, the Subject method includes the use of sirtuin-modu other dosage form is a Sustained release form which is Suit lating compounds that increase the level and/or activity of a able for administration two to four times per day. In contrast sirtuin protein, alone or in combination with other agents, for to IR formulations, sustained release (“SR) nicotinic acid reducing incidence or severity of flushing and/or hot flashes formulations are designed to release significant quantities of in cancer patients. In other embodiments, the method pro drug for absorption into the blood stream over specific timed vides for the use of sirtuin-modulating compounds that intervals in order to maintain therapeutic levels of nicotinic increase the level and/or activity of a sirtuin protein to reduce acid over an extended period such as 12 or 24 hours after the incidence or severity of flushing and/or hot flashes in ingestion. menopausal and post-menopausal woman. 0373. As used herein, the term “nicotinic acid' is meant to 0369. In another aspect, sirtuin-modulating compounds encompass nicotinic acid or a compound other than nicotinic that increase the level and/or activity of a sirtuin protein may acid itself which the body metabolizes into nicotinic acid, be used as a therapy for reducing the incidence or severity of thus producing essentially the same effect as nicotinic acid. flushing and/or hot flashes which are side-effects of another Exemplary compounds that produce an effect similar to that drug therapy, e.g., drug-induced flushing. In certain embodi of nicotinic acid include, for example, nicotinyl alcohol tar ments, a method for treating and/or preventing drug-induced trate, d-glucitol hexanicotinate, aluminum nicotinate, nicer flushing comprises administering to a patient in need thereof itrol and d. 1-alpha-tocopheryl nicotinate. Each Such com a formulation comprising at least one flushing inducing com pound will be collectively referred to herein as “nicotinic pound and at least one sirtuin-modulating compound that acid.” increases the level and/or activity of a sirtuin protein. In other 0374. In another embodiment, the invention provides a embodiments, a method for treating drug induced flushing method for treating and/or preventing hyperlipidemia with comprises separately administering one or more compounds reduced flushing side effects. The method comprises the steps that induce flushing and one or more sirtuin-modulating com of administering to a subject in need thereofatherapeutically pounds, e.g., wherein the sirtuin-modulating compound and effective amount of nicotinic acid and a sirtuin-modulating flushing inducing agent have not been formulated in the same compound that increases the level and/or activity of a sirtuin compositions. When using separate formulations, the sirtuin protein in an amount Sufficient to reduce flushing. In an exem US 2009/0143376 A1 Jun. 4, 2009 37 plary embodiment, the nicotinic acid and/or sirtuin-modulat tor, such as selected from the group consisting of isocarbox ing compound may be administered nocturnally. azid, phenelZine, tranylcypromine, selegiline and moclobe 0375. In another representative embodiment, the method mide. involves the use of Sirtuin-modulating compounds that 0378. In still another representative embodiment, sirtuin increase the level and/or activity of a sirtuin protein to reduce modulating compounds that increase the level and/or activity flushing side effects of raloxifene. Raloxifene acts like estro of a sirtuin protein may be used to reduce flushing side effects gen in certain places in the body, but is not a hormone. It helps of chemotherapeutic agents, such as cyclophosphamide, prevent osteoporosis in women who have reached meno tamoxifen. pause. Osteoporosis causes bones to gradually grow thin, 0379. In another embodiment, sirtuin-modulating com fragile, and more likely to break. Evista slows down the loss pounds that increase the level and/or activity of a sirtuin of bone mass that occurs with menopause, lowering the risk of protein may be used to reduce flushing side effects of calcium spine fractures due to osteoporosis. A common side effect of channel blockers, such as amlodipine. raloxifene is hot flashes (Sweating and flushing). This can be 0380. In another embodiment, sirtuin-modulating com pounds that increase the level and/or activity of a sirtuin uncomfortable for women who already have hot flashes due protein may be used to reduce flushing side effects of antibi to menopause. otics. For example, sirtuin-modulating compounds that 0376. In another representative embodiment, the method increase the level and/or activity of a sirtuin protein can be involves the use of Sirtuin-modulating compounds that used in combination with levofloxacin. Levofloxacin is used increase the level and/or activity of a sirtuin protein to reduce to treat infections of the sinuses, skin, lungs, ears, airways, flushing side effects of antidepressants or anti-psychotic bones, and joints caused by Susceptible bacteria. Levofloxa agent. For instance, sirtuin-modulating compounds that cin also is frequently used to treaturinary infections, includ increase the level and/or activity of a sirtuin protein can be ing those resistant to other antibiotics, as well as prostatitis. used in conjunction (administered separately or together) Levofloxacin is effective in treating infectious diarrheas with a serotonin reuptake inhibitor, a 5HT2 receptor antago caused by E. coli, campylobacter jejuni, and Shigella bacte nist, an anticonvulsant, a norepinephrine reuptake inhibitor, ria. Levofloxacin also can be used to treat various obstetric an C.-adrenoreceptor antagonist, an NK-3 antagonist, an infections, including mastitis. NK-1 receptor antagonist, a PDE4 inhibitor, an Neuropeptide Y5 Receptor Antagonists, a D4 receptor antagonist, a 5HT1A Ocular Disorders receptor antagonist, a 5HT1D receptor antagonist, a CRF 0381. One aspect of the present invention is a method for antagonist, a monoamine oxidase inhibitor, or a sedative inhibiting, reducing or otherwise treating vision impairment hypnotic drug. by administering to a patient a therapeutic dosage of sirtuin 0377. In certain embodiments, sirtuin-modulating com modulator selected from a compound disclosed herein, or a pounds that increase the level and/or activity of a sirtuin pharmaceutically acceptable salt, prodrug or a metabolic protein may be used as part of a treatment with a serotonin derivative thereof. reuptake inhibitor (SRI) to reduce flushing. In certain pre 0382. In certain aspects of the invention, the vision impair ferred embodiments, the SRI is a selective serotonin reuptake ment is caused by damage to the optic nerve or central ner inhibitor (SSRI), such as a fluoxetinoid (fluoxetine, norflu Vous system. In particular embodiments, optic nerve damage oxetine) or a nefazodonoid (nefazodone, hydroxynefaz is caused by high intraocular pressure. Such as that created by odone, oxonefazodone). Other exemplary SSRI's include glaucoma. In other particular embodiments, optic nerve dam dulloxetine, Venlafaxine, milnacipran, citalopram, fluvoxam age is caused by Swelling of the nerve, which is often asso ine, paroxetine and Sertraline. The sirtuin-modulating com ciated with an infection or an immune (e.g., autoimmune) pound that increases the level and/or activity of a sirtuin response Such as in optic neuritis. protein can also be used as part of a treatment with sedative 0383 Glaucoma describes a group of disorders which are hypnotic drug, Such as selected from the group consisting of associated with a visual field defect, cupping of the optic disc, a benzodiazepine (Such as alprazolam, chlordiazepoxide, and optic nerve damage. These are commonly referred to as clonazepam, chloraZepate, clobazam, diazepam, halazepam, glaucomatous optic neuropathies. Most glaucomas are usu lorazepam, oxazepam and prazepam), Zolpidem, and barbi ally, but not always, associated with a rise in intraocular turates. In still other embodiments, a sirtuin-modulating com pressure. Exemplary forms of glaucoma include Glaucoma pound that increases the level and/or activity of a sirtuin and Penetrating Keratoplasty, Acute Angle Closure, Chronic protein may be used as part of a treatment with a 5-HT1A Angle Closure, Chronic Open Angle, Angle Recession, Aph receptor partial agonist, such as selected from the group con akic and Pseudophakic, Drug-Induced, Hyphema, Intraocu sisting of buspirone, flesinoxan, gepirone and ipsapirone. lar Tumors, Juvenile, Lens-Particle, Low Tension, Malignant, Sirtuin-modulating compounds that increase the level and/or Neovascular, Phacolytic, Phacomorphic, Pigmentary, Pla activity of a sirtuin protein can also used as part of a treatment teau Iris, Primary Congenital, Primary Open Angle, Pseu with a norepinephrine reuptake inhibitor, Such as selected doexfoliation, Secondary Congenital, Adult Suspect, Unilat from tertiary amine tricyclics and secondary amine tricyclics. eral, Uveitic, Ocular Hypertension, Ocular Hypotony, Exemplary tertiary amine tricyclic include amitriptyline, clo Posner-Schlossman Syndrome and Scleral Expansion Proce mipramine, doxepin, imipramine and trimipramine. Exem dure in Ocular Hypertension & Primary Open-angle Glau plary secondary amine tricyclic include amoxapine, COa. desipramine, maprotiline, nortriptyline and protriptyline. In 0384 Intraocular pressure can also be increased by vari certain embodiments, sirtuin-modulating compounds that ous Surgical procedures, such as phacoemulsification (i.e., increase the level and/or activity of a sirtuin protein may be cataract Surgery) and implantation of structures such as an used as part of a treatment with a monoamine oxidase inhibi artificial lens. In addition, spinal Surgeries in particular, or any US 2009/0143376 A1 Jun. 4, 2009

Surgery in which the patient is prone for an extended period of lis, gonorrhea), viral infections (e.g. Ocular Herpes Simplex time can lead to increased interocular pressure. Virus, Varicella Zoster Virus, Cytomegalovirus retinitis, 0385 Optic neuritis (ON) is inflammation of the optic Human Immunodeficiency Virus (HIV)) as well as progres nerve and causes acute loss of vision. It is highly associated sive outer retinal necrosis secondary to HIV or other HIV with multiple sclerosis (MS) as 15-25% of MS patients ini associated and other immunodeficiency-associated ocular tially present with ON, and 50-75% of ON patients are diag diseases. In addition, ocular diseases include fungal infec nosed with MS. ON is also associated with infection (e.g., tions (e.g. Candida choroiditis, histoplasmosis), protozoal viral infection, meningitis, syphilis), inflammation (e.g., infections (e.g. toxoplasmosis) and others such as ocular from a vaccine), infiltration and ischemia. toxocariasis and sarcoidosis. 0386. Another condition leading to optic nerve damage is One aspect of the invention is a method for inhibiting, reduc anterior ischemic optic neuropathy (AION). There are two ing or treating vision impairment in a subject undergoing types of AION. Arteritic AION is due to giant cell arteritis treatment with a chemotherapeutic drug (e.g., a neurotoxic (vasculitis) and leads to acute vision loss. Non-arteritic AION drug, a drug that raises intraocular pressure Such as a steroid), encompasses all cases of ischemic optic neuropathy other by administering to the Subject in need of Such treatment a than those due to giant cell arteritis. The pathophysiology of therapeutic dosage of a sirtuin modulator disclosed herein. AION is unclear although it appears to incorporate both 0392 Another aspect of the invention is a method for inflammatory and ischemic mechanisms. inhibiting, reducing or treating vision impairment in a subject 0387. Other damage to the optic nerve is typically associ undergoing Surgery, including ocular or other Surgeries per ated with demyleination, inflammation, ischemia, toxins, or formed in the prone position Such as spinal cord Surgery, by trauma to the optic nerve. Exemplary conditions where the administering to the Subject in need of such treatmentathera optic nerve is damaged include Demyelinating Optic Neur peutic dosage of a sirtuin modulator disclosed herein. Ocular opathy (Optic Neuritis, Retrobulbar Optic Neuritis), Optic Surgeries include cataract, iridotomy and lens replacements. Nerve Sheath Meningioma, Adult Optic Neuritis, Childhood Another aspect of the invention is the treatment, including Optic Neuritis, Anterior Ischemic Optic Neuropathy, Poste inhibition and prophylactic treatment, of age related ocular rior Ischemic Optic Neuropathy, Compressive Optic Neur diseases include cataracts, dry eye, retinal damage and the opathy, Papilledema, Pseudopapilledema and Toxic/Nutri like, by administering to the Subject in need of such treatment tional Optic Neuropathy. a therapeutic dosage of a sirtuin modulator disclosed herein. 0388 Other neurological conditions associated with 0393. The formation of cataracts is associated with several vision loss, albeit not directly associated with damage to the biochemical changes in the lens of the eye, such as decreased optic nerve, include Amblyopia, Bells Palsy, Chronic Pro levels of antioxidants ascorbic acid and glutathione, gressive External Ophthalmoplegia, Multiple Sclerosis, increased lipid, amino acid and protein oxidation, increased Pseudotumor Cerebri and Trigeminal Neuralgia. Sodium and calcium, loss of amino acids and decreased lens 0389. In certain aspects of the invention, the vision impair metabolism. The lens, which lacks blood vessels, is sus ment is caused by retinal damage. In particular embodiments, pended in extracellular fluids in the anterior part of the eye. retinal damage is caused by disturbances in blood flow to the Nutrients, such as ascorbic acid, glutathione, Vitamin E, sele eye (e.g., arteriosclerosis, vasculitis). In particular embodi nium, bioflavonoids and carotenoids are required to maintain ments, retinal damage is caused by disruption of the macula the transparency of the lens. Low levels of selenium results in (e.g., exudative or non-exudative macular degeneration). an increase of free radical-inducing hydrogen peroxide, 0390 Exemplary retinal diseases include Exudative Age which is neutralized by the selenium-dependent antioxidant Related Macular Degeneration, Nonexudative Age Related enzyme glutathione peroxidase. Lens-protective glutathione Macular Degeneration, Retinal Electronic Prosthesis and peroxidase is also dependent on the amino acids methionine, RPE Transplantation Age Related Macular Degeneration, cysteine, glycine and glutamic acid. Acute Multifocal Placoid Pigment Epitheliopathy, Acute 0394 Cataracts can also develop due to an inability to Retinal Necrosis, Best Disease, Branch Retinal Artery Occlu properly metabolize galactose found in dairy products that sion, Branch Retinal Vein Occlusion, Cancer Associated and contain lactose, a disaccharide composed of the monosaccha Related Autoimmune Retinopathies, Central Retinal Artery ride galactose and glucose. Cataracts can be prevented, Occlusion, Central Retinal Vein Occlusion, Central Serous delayed, slowed and possibly even reversed if detected early Chorioretinopathy, Eales Disease, Epimacular Membrane, and metabolically corrected. Lattice Degeneration, Macroaneurysm, Diabetic Macular 0395. Retinal damage is attributed, interalia, to free radi Edema, Irvine-Gass Macular Edema, Macular Hole, Subreti cal initiated reactions in glaucoma, diabetic retinopathy and nal Neovascular Membranes, Diffuse Unilateral Subacute age-related macular degeneration (AMD). The eye is a part of Neuroretinitis, Nonpseudophakic Cystoid Macular Edema, the central nervous system and has limited regenerative capa Presumed Ocular Histoplasmosis Syndrome. Exudative Reti bility. The retina is composed of numerous nerve cells which nal Detachment, Postoperative Retinal Detachment, Prolif contain the highest concentration of polyunsaturated fatty erative Retinal Detachment, Rhegmatogenous Retinal acids (PFA) and subject to oxidation. Free radicals are gen Detachment, Tractional Retinal Detachment, Retinitis Pig erated by UV light entering the eye and mitochondria in the mentosa, CMV Retinitis, Retinoblastoma, Retinopathy of rods and cones, which generate the energy necessary to trans Prematurity, Birdshot Retinopathy, Background Diabetic form light into visual impulses. Free radicals cause peroxi Retinopathy, Proliferative Diabetic Retinopathy, Hemoglo dation of the PFA by hydroxyl or superoxide radicals which in binopathies Retinopathy, Purtscher Retinopathy, Valsalva turn propagate additional free radicals. The free radicals Retinopathy, Juvenile Retinoschisis, Senile Retinoschisis, cause temporary or permanent damage to retinal tissue. Terson Syndrome and White Dot Syndromes. 0396 Glaucoma is usually viewed as a disorder that 0391. Other exemplary diseases include ocular bacterial causes an elevated intraocular pressure (IOP) that results in infections (e.g. conjunctivitis, keratitis, tuberculosis, syphi permanent damage to the retinal nerve fibers, but a sixth of all US 2009/0143376 A1 Jun. 4, 2009 39 glaucoma cases do not develop an elevated IOP. This disorder Such treatment a therapeutic dosage of a sirtuin modulator is now perceived as one of reduced vascular perfusion and an disclosed herein. Radiation or electromagnetic damage to the increase in neurotoxic factors. Recent studies have implicated eye can include that caused by CRT's or exposure to sunlight elevated levels of glutamate, nitric oxide and peroxynitirite in or UV. the eye as the causes of the death of retinal ganglion cells. Neuroprotective agents may be the future of glaucoma care. 0402. In one embodiment, a combination drug regimen For example, nitric oxide synthase inhibitors block the for may include drugs or compounds for the treatment or preven mation of peroxynitrite from nitric oxide and Superoxide. In a tion of ocular disorders or secondary conditions associated recent study, animals treated with aminoguanidine, a nitric with these conditions. Thus, a combination drug regimen may oxide synthase inhibitor, had a reduction in the loss of retinal include one or more sirtuin activators and one or more thera ganglion cells. It was concluded that nitric oxide in the eye peutic agents for the treatment of an ocular disorder. For caused cytotoxicity in many tissues and neurotoxicity in the example, one or more sirtuin-activating compounds can be central nervous system. combined with an effective amount of one or more of: an 0397 Diabetic retinopathy occurs when the underlying agent that reduces intraocular pressure, an agent for treating blood vessels develop microvascular abnormalities consist glaucoma, an agent for treating optic neuritis, an agent for ing primarily of microaneurysms and intraretinal hemor treating CMV Retinopathy, an agent for treating multiple rhages. Oxidative metabolites are directly involved with the Sclerosis, and/or an antibiotic, etc. pathogenesis of diabetic retinopathy and free radicals aug 0403. In one embodiment, a sirtuin modulator can be ment the generation of growth factors that lead to enhanced administered in conjunction with a therapy for reducing proliferative activity. Nitric oxide produced by endothelial intraocular pressure. One group of therapies involves block cells of the vessels may also cause Smooth muscle cells to ing aqueous production. For example, topical beta-adrenergic relax and result in vasodilation of segments of the vessel. antagonists (timolol and betaxolol) decrease aqueous produc Ischemia and hypoxia of the retina occur after thickening of tion. Topical timolol causes IOP to fall in 30 minutes with the arterial basement membrane, endothelial proliferation peak effects in 1-2 hours. A reasonable regimen is Timoptic and loss of pericytes. The inadequate oxygenation causes 0.5%, one drop every 30 minutes for 2 doses. The carbonic capillary obliteration or nonperfusion, arteriolar-Venular anhydrase inhibitor, acetazolamide, also decreases aqueous shunts, sluggish blood flow and an impaired ability of RBCs production and should be given in conjunction with topical to release oxygen. Lipid peroxidation of the retinal tissues beta-antagonists. An initial dose of 500 mg is administered also occurs as a result of free radical damage. followed by 250 mg every 6 hours. This medication may be 0398. The macula is responsible for our acute central given orally, intramuscularly, or intravenously. In addition, vision and composed of light-sensing cells (cones) while the alpha 2-agonists (e.g., Apraclonidine) act by decreasing underlying retinal pigment epithelium (RPE) and choroid aqueous production. Their effects are additive to topically nourish and help remove waste materials. The RPE nourishes administered beta-blockers. They have been approved for use the cones with the vitamin A substrate for the photosensitive in controlling an acute rise in pressure following anterior pigments and digests the cones shed outer tips. RPE is chamber laser procedures, but has been reported effective in exposed to high levels of UV radiation, and secretes factors treating acute closed-angle glaucoma. A reasonable regimen that inhibit angiogenesis. The choroid contains a dense vas is 1 drop every 30 minutes for 2 doses. cular network that provides nutrients and removes the waste 0404 A second group of therapies for reducing intraocular materials. pressure involve reducing vitreous Volume. Hyperosmotic 0399. In AMD, the shed cone tips become indigestible by agents can be used to treat an acute attack. These agents draw the RPE, where the cells Swell and die after collecting too water out of the globe by making the blood hyperosmolar. much undigested material. Collections of undigested waste Oral glycerol in a dose of 1 mL/g in a cold 50% solution material, called drusen, form under the RPE. Photoxic dam (mixed with lemon juice to make it more palatable) often is age also causes the accumulation of lipofuscin in RPE cells. used. Glycerol is converted to glucose in the liver, persons The intracellular lipofuscin and accumulation of drusen in with diabetes may need additional insulin if they become Bruch's membrane interferes with the transport of oxygen hyperglycemic after receiving glycerol. Oral isosorbide is a and nutrients to the retinal tissues, and ultimately leads to metabolically inert alcohol that also can be used as an osmotic RPE and photoreceptor dysfunction. In exudative AMD, agent for patients with acute angle-closure glaucoma. Usual blood vessels grow from the choriocapillaris through defects dose is 100 g taken po. (220 cc of a 45% solution). This inert in Bruch's membrane and may grow under the RPE, detach alcohol should not be confused with isosorbide dinitrate, a ing it from the choroid, and leaking fluid or bleeding. nitrate-based cardiac medication used forangina and for con 0400 Macular pigment, one of the protective factors that gestive heart failure. Intravenous mannitol in a dose of 1.0-1.5 prevent Sunlight from damaging the retina, is formed by the mg/kg also is effective and is well tolerated in patients with accumulation of nutritionally derived carotenoids, such as nausea and vomiting. These hyperosmotic agents should be lutein, the fatty yellow pigment that serves as a delivery used with caution in any patient with a history of congestive vehicle for other important nutrients and Zeaxanthin. Antioxi heart failure. dants such as vitamins C and E, beta-carotene and lutein, as 0405. A third group of therapies involve facilitating aque well as Zinc, Selenium and copper, are all found in the healthy ous outflow from the eye. Miotic agents pull the iris from the macula. In addition to providing nourishment, these antioxi iridocorneal angle and may help to relieve the obstruction of dants protect against free radical damage that initiates macu the trabecular meshwork by the peripheral iris. Pilocarpine lar degeneration. 2% (blue eyes)-4% (brown eyes) can be administered every 04.01. Another aspect of the invention is the prevention or 15 minutes for the first 1-2 hours. More frequent administra treatment of damage to the eye caused by stress, chemical tion or higher doses may precipitate a systemic cholinergic insult or radiation, by administering to the Subject in need of crisis. NSAIDS are sometimes used to reduce inflammation. US 2009/0143376 A1 Jun. 4, 2009 40

04.06 Exemplary therapeutic agents for reducing mitochondria while maintaining the overall numbers of mito intraocular pressure include ALPHAGANR P (Allergan) chondria (e.g., mitochondrial mass), increasing the numbers (brimonidine tartrate ophthalmic solution), AZOPTR (Al of mitochondria thereby increasing mitochondrial activity con) (brinzolamide ophthalmic suspension), BETAGANR) (e.g., by Stimulating mitochondrial biogenesis), or combina (Allergan) (levobunolol hydrochloride ophthalmic solution, tions thereof. In certain embodiments, diseases and disorders USP), BETIMOL(R) (Vistakon) (timolol ophthalmic solu that would benefit from increased mitochondrial activity tion), BETOPTICS(R) (Alcon) (betaxolol HCl), BRIMONI include diseases or disorders associated with mitochondrial DINE TARTRATE (Bausch & Lomb), CARTEOLOL dysfunction. HYDROCHLORIDE (Bausch & Lomb), COSOPTR 0413. In certain embodiments, methods for treating dis (Merck) (dorzolamide hydrochloride-timolol maleate oph eases or disorders that would benefit from increased mito thalmic solution), LUMIGANR) (Allergan) (bimatoprost chondrial activity may comprise identifying a Subject Suffer ophthalmic solution), OPTIPRANOLOL(R) (Bausch & ing from a mitochondrial dysfunction. Methods for Lomb) (metipranolol ophthalmic solution), TIMOLOL GFS diagnosing a mitochondrial dysfunction may involve molecu (Falcon) (timolol maleate ophthalmic gel forming solution), lar genetic, pathologic and/or biochemical analysis are sum TIMOPTICR (Merck) (timolol maleate ophthalmic solu marized in Cohen and Gold, Cleveland Clinic Journal of tion), TRAVATANR) (Alcon) (travoprost ophthalmic solu Medicine, 68: 625-642 (2001). One method for diagnosing a tion), TRUSOPTR (Merck) (dorzolamide hydrochloride mitochondrial dysfunction is the Thor-Byme-ier scale (see ophthalmic solution) and XALATANR) (Pharmacia & e.g., Cohen and Gold, supra; Collin S. et al., Eur Neurol. 36: Upjohn) (latanoprost ophthalmic solution). 260-267 (1996)). Other methods for determining mitochon 0407. In one embodiment, a sirtuin modulator can be drial number and function include, for example, enzymatic administered in conjunction with a therapy for treating and/or assays (e.g., a mitochondrial enzyme or an ATP biosynthesis preventing glaucoma. An example of a glaucoma drug is factor Such as an ETC enzyme or a Krebs cycle enzyme), DARANIDE(R) Tablets (Merck) (Dichlorphenamide). determination or mitochondrial mass, mitochondrial Volume, 0408. In one embodiment, a sirtuin modulator can be and/or mitochondrial number, quantification of mitochon administered in conjunction with a therapy for treating and/or drial DNA, monitoring intracellular calcium homeostasis preventing optic neuritis. Examples of drugs for optic neuritis and/or cellular responses to perturbations of this homeostasis, include DECADRONR Phosphate Injection (Merck) (Dex evaluation of response to an apoptogenic stimulus, determi amethasone Sodium Phosphate), DEPO-MEDROL(R) (Phar nation of free radical production. Such methods are known in macia & Upjohn)(methylprednisolone acetate), HYDRO the art and are described, for example, in U.S. Patent Publi CORTONER) Tablets (Merck) (Hydrocortisone), cation No. 2002/0049176 and references cited therein. ORAPREDR) (Biomarin) (prednisolone sodium phosphate 0414. Mitochondria are critical for the survival and proper oral solution) and PEDIAPREDR (Celltech) (prednisolone function of almost all types of eukaryotic cells. Mitochondria sodium phosphate, USP). in virtually any cell type can have congenital or acquired 04.09. In one embodiment, a sirtuin modulator can be defects that affect their function. Thus, the clinically signifi administered in conjunction with a therapy for treating and/or cant signs and symptoms of mitochondrial defects affecting preventing CMV Retinopathy. Treatments for CMV retinopa respiratory chain function are heterogeneous and variable thy include CYTOVENER) (ganciclovir capsules) and VAL depending on the distribution of defective mitochondria CYTER (Roche Laboratories) (valganciclovir hydrochloride among cells and the severity of their deficits, and upon physi tablets). ological demands upon the affected cells. Nondividing tis 0410. In one embodiment, a sirtuin modulator can be Sues with high energy requirements, e.g. nervous tissue, skel administered in conjunction with a therapy for treating and/or etal muscle and cardiac muscle are particularly Susceptible to preventing multiple Sclerosis. Examples of Such drugs mitochondrial respiratory chain dysfunction, but any organ include DANTRIUM(R) (Procter & Gamble Pharmaceuticals) system can be affected. (dantrolene sodium), NOVANTRONER) (Serono) (mitox 0415 Diseases and disorders associated with mitochon antrone), AVONEXR (Biogen Idec) (Interferon beta-1a). drial dysfunction include diseases and disorders in which BETASERONR) (Berlex) (Interferon beta-1b), COPAX deficits in mitochondrial respiratory chain activity contribute ONER) (Teva Neuroscience) (glatiramer acetate injection) to the development of pathophysiology of Such diseases or and REBIFR (Pfizer) (interferon beta-1a). disorders in a mammal. This includes 1) congenital genetic 0411. In addition, macrollide and/or mycophenolic acid, deficiencies in activity of one or more components of the which has multiple activities, can be co-administered with a mitochondrial respiratory chain; and 2) acquired deficiencies sirtuin modulator. Macrollide antibiotics include tacrolimus, in the activity of one or more components of the mitochon cyclosporine, Sirolimus, everolimus, ascomycin, erythromy drial respiratory chain, wherein such deficiencies are caused cin, azithromycin, clarithromycin, clindamycin, lincomycin, by a) oxidative damage during aging; b) elevated intracellular dirithromycin, josamycin, spiramycin, diacetyl-midecamy calcium; c) exposure of affected cells to nitric oxide; d) cin, tylosin, roxithromycin, ABT-773, tellithromycin, leuco hypoxia or ischemia; e) microtubule-associated deficits in mycins, and lincosamide. axonal transport of mitochondria, or f) expression of mito chondrial uncoupling proteins. Mitochondrial-Associated Diseases and Disorders 0416 Diseases or disorders that would benefit from 0412. In certain embodiments, the invention provides increased mitochondrial activity generally include for methods for treating diseases or disorders that would benefit example, diseases in which free radical mediated oxidative from increased mitochondrial activity. The methods involve injury leads to tissue degeneration, diseases in which cells administering to a Subject in need thereof a therapeutically inappropriately undergo apoptosis, and diseases in which effective amount of a sirtuin activating compound. Increased cells fail to undergo apoptosis. Exemplary diseases or disor mitochondrial activity refers to increasing activity of the ders that would benefit from increased mitochondrial activity US 2009/0143376 A1 Jun. 4, 2009

include, for example, AD (Alzheimer's Disease), ADPD thrombocytopenia and leukemia syndrome, MARIAHS syn (Alzheimer's Disease and Parkinsons's Disease). AMDF drome (Mitrochondrial ataxia, recurrent infections, aphasia, (Ataxia, Myoclonus and Deafness), auto-immune disease, hypouricemia/hypomyelination, seizures, and dicarboxylic cancer, CIPO (Chronic Intestinal Pseudoobstruction with aciduria), ND6 dystonia, Cyclic vomiting syndrome with myopathy and Opthalmoplegia), congenital muscular dystro declines during infection, 3-Hydroxyisobutryic aciduria with phy, CPEO (Chronic Progressive External Opthalmoplegia), lactic acidemia, Diabetes mellitus with lactic acidemia, Uri DEAF (Maternally inherited DEAFness or aminoglycoside dine responsive neurologic syndrome (URNS), Dilated car induced DEAFness), DEMCHO (Dementia and Chorea), dia diomyopathy, Splenic Lymphoma, and Renal Tubular Acido betes mellitis (Type I or Type II), DIDMOAD (Diabetes sis/Diabetes/Ataxis syndrome. Insipidus, Diabetes Mellitus, Optic Atrophy, Deafness), 0418. In other embodiments, the invention provides meth DMDF (Diabetes Mellitus and Deafness), dystonia, Exercise ods for treating a Subject Suffering from mitochondrial disor Intolerance, ESOC (Epilepsy, Strokes, Optic atrophy, and ders arising from, but not limited to, post-traumatic head Cognitive decline), FBSN (Familial Bilateral Striatal Necro injury and cerebral edema, stroke (invention methods useful sis), FICP (Fatal Infantile Cardiomyopathy Plus, a MELAS for preventing or preventing reperfusion injury), Lewy body associated cardiomyopathy), GER (Gastrointestinal Reflux), dementia, hepatorenal syndrome, acute liver failure, NASH HD (Huntington's Disease), KSS (Kearns Sayre Syndrome), (non-alcoholic steatohepatitis), Anti-metastasis/prodifferen “later-onset myopathy, LDYT (Leber's hereditary optic neu tiation therapy of cancer, idiopathic congestive heart failure, ropathy and DYsTonia), Leigh's Syndrome, LHON (Leber atrial fibrilation (non-valvular), Wolff-Parkinson-White Syn Hereditary Optic Neuropathy), LIMM (Lethal Infantile Mito drome, idiopathic heart block, prevention of reperfusion chondrial Myopathy), MDM (Myopathy and Diabetes Mel injury in acute myocardial infarctions, familial migraines, litus), MELAS (Mitochondrial Encephalomyopathy, Lactic irritable bowel syndrome, secondary prevention of non-Q Acidosis, and Stroke-like episodes), MEPR (Myoclonic Epi wave myocardial infarctions, Premenstrual syndrome, Pre lepsy and Psychomotor Regression), MERME (MERRF/ vention of renal failure in hepatorenal syndrome, anti-phos MELAS overlap disease), MERRF (Myoclonic Epilepsy and pholipid antibody syndrome, eclampsia/pre-eclampsia, Ragged Red Muscle Fibers), MHCM (Maternally Inherited oopause infertility, ischemic heart disease/angina, and Shy Hypertrophic CardioMyopathy), MICM (Maternally Inher Drager and unclassified dysautonomia syndromes. ited Cardiomyopathy), MILS (Maternally Inherited Leigh 0419. In still another embodiment, there are provided Syndrome), Mitochondrial Encephalocardiomyopathy, methods for the treatment of mitochondrial disorders associ Mitochondrial Encephalomyopathy, MM (Mitochondrial ated with pharmacological drug-related side effects. Types of Myopathy), MMC (Maternal Myopathy and Cardiomyopa pharmaceutical agents that are associated with mitochondrial thy), MNGIE (Myopathy and external ophthalmoplegia, disorders include reverse transcriptase inhibitors, protease Neuropathy, Gastro-Intestinal, Encephalopathy), Multisys inhibitors, inhibitors of DHOD, and the like. Examples of tem. Mitochondrial Disorder (myopathy, encephalopathy, reverse transcriptase inhibitors include, for example, AZidot blindness, hearing loss, peripheral neuropathy), NARP (Neu hymidine (AZT), Stavudine (D4T), Zalcitabine (ddC). rogenic muscle weakness, Ataxia, and Retinitis Pigmentosa; Didanosine (DDI), Fluoroiodoarauracil (FIAU), Lamivudine alternate phenotype at this locus is reported as Leigh Dis (3TC), Abacavir and the like. Examples of protease inhibitors ease), PD (Parkinson's Disease), Pearson's Syndrome, PEM include, for example, Ritonavir, Indinavir, Saquinavir, Nelfi (Progressive Encephalopathy), PEO (Progressive External navir and the like. Examples of inhibitors of dihydroorotate Opthalmoplegia), PME (Progressive Myoclonus Epilepsy), dehydrogenase (DHOD) include, for example, Leflunomide, PMPS (Pearson Marrow-Pancreas Syndrome), psoriasis, Brequinar, and the like. RTT (Rett Syndrome), schizophrenia, SIDS (Sudden Infant 0420 Reverse transcriptase inhibitors not only inhibit Death Syndrome), SNHL (Sensorineural Hearing Loss), Var reverse transcriptase but also polymerase gamma which is ied Familial Presentation (clinical manifestations range from required for mitochondrial function. Inhibition of polymerase spastic paraparesis to multisystem progressive disorder & gamma activity (e.g., with a reverse transcriptase inhibitor) fatal cardiomyopathy to truncal ataxia, dysarthria, severe therefore leads to mitochondrial dysfunction and/or a reduced hearing loss, mental regression, ptosis, ophthalmoparesis, mitochondrial mass which manifests itself in patients as distal cyclones, and diabetes mellitus), or Wolfram syndrome. hyperlactatemia. This type of condition may benefit from an 0417. Other diseases and disorders that would benefit increase in the number of mitochondria and/or an improve from increased mitochondrial activity include, for example, ment in mitochondrial function, e.g., by administration of a Friedreich's ataxia and other ataxias, amyotrophic lateral sirtuin activating compound. Sclerosis (ALS) and other motor neuron diseases, macular 0421 Common symptoms of mitochondrial diseases degeneration, epilepsy, Alpers syndrome, Multiple mito include cardiomyopathy, muscle weakness and atrophy, chondrial DNA deletion syndrome, MTDNA depletion syn developmental delays (involving motor, language, cognitive drome, Complex I deficiency, Complex II (SDH) deficiency, or executive function), ataxia, epilepsy, renal tubular acidosis, Complex III deficiency, Cytochrome c oxidase (COX, Com peripheral neuropathy, optic neuropathy, autonomic neuropa plex IV) deficiency, Complex V deficiency, Adenine Nucle thy, neurogenic bowel dysfunction, sensorineural deafness, otide Translocator (ANT) deficiency, Pyruvate dehydroge neurogenic bladder dysfunction, dilating cardiomyopathy, nase (PDH) deficiency, Ethylmalonic aciduria with lactic migraine, hepatic failure, lactic acidemia, and diabetes mel acidemia, 3-Methylglutaconic aciduria with lactic acidemia, litus. Refractory epilepsy with declines during infection, Asperger 0422. In certain embodiments, the invention provides syndrome with declines during infection, Autism with methods for treating a disease or disorder that would benefit declines during infection, Attention deficit hyperactivity dis from increased mitochondrial activity that involves adminis order (ADHD), Cerebral palsy with declines during infection, tering to a subject in need thereof one or more sirtuin activat Dyslexia with declines during infection, materially inherited ing compounds in combination with another therapeutic US 2009/0143376 A1 Jun. 4, 2009 42 agent such as, for example, an agent useful for treating mito tion, often resulting in atrophy of skeletal muscle and myo chondrial dysfunction (Such as antioxidants, vitamins, or res cardial dysfunction. In the case of Duchenne muscular dys piratory chain cofactors), an agent useful for reducing a trophy, mutations or deficits in a specific protein, dystrophin, symptom associated with a disease or disorder involving are implicated in its etiology. Mice with their dystrophin mitochondrial dysfunction (Such as, an anti-seizure agent, an genes inactivated display some characteristics of muscular agent useful for alleviating neuropathic pain, an agent for dystrophy, and have an approximately 50% deficit in mito treating cardiac dysfunction), a cardiovascular agent (as chondrial respiratory chain activity. A final common pathway described further below), a chemotherapeutic agent (as for neuromuscular degeneration in most cases is calcium described further below), or an anti-neurodegeneration agent mediated impairment of mitochondrial function. In certain (as described further below). In an exemplary embodiment, embodiments, sirtuin activating compounds may be used for the invention provides methods for treating a disease or dis reducing the rate of decline in muscular functional capacities order that would benefit from increased mitochondrial activ and for improving muscular functional status in patients with ity that involves administering to a subject in need thereofone muscular dystrophy. or more sirtuin activating compounds in combination with 0426 Multiple sclerosis (MS) is a neuromuscular disease one or more of the following: coenzyme Qo, L-carnitine, characterized by focal inflammatory and autoimmune degen thiamine, riboflavin, niacinamide, folate, vitamin E, sele eration of cerebral white matter. Periodic exacerbations or nium, lipoic acid, or prednisone. Compositions comprising attacks are significantly correlated with upper respiratory Such combinations are also provided herein. tract and other infections, both bacterial and viral, indicating 0423. In exemplary embodiments, the invention provides that mitochondrial dysfunction plays a role in MS. Depres methods for treating diseases or disorders that would benefit sion of neuronal mitochondrial respiratory chain activity from increased mitochondrial activity by administering to a caused by Nitric Oxide (produced by astrocytes and other Subject a therapeutically effective amount of a sirtuin activat cells involved in inflammation) is implicated as a molecular ing compound. Exemplary diseases or disorders include, for mechanism contributing to MS. In certain embodiments, sir example, neuromuscular disorders (e.g., Friedreich's Ataxia, tuin activating compounds may be used for treatment of muscular dystrophy, multiple Sclerosis, etc.), disorders of patients with multiple Sclerosis, both prophylactically and neuronal instability (e.g., seizure disorders, migraine, etc.), during episodes of disease exacerbation. developmental delay, neurodegenerative disorders (e.g., 0427 Epilepsy is often present in patients with mitochon Alzheimer's Disease, Parkinson's Disease, amyotrophic lat drial cytopathies, involving a range of seizure severity and eral sclerosis, etc.), ischemia, renal tubular acidosis, age frequency, e.g. absence, tonic, atonic, myoclonic, and status related neurodegeneration and cognitive decline, chemo epilepticus, occurring in isolated episodes or many times therapy fatigue, age-related or chemotherapy-induced daily. In certain embodiments, sirtuin activating compounds menopause or irregularities of menstrual cycling or ovula may be used for treating patients with seizures secondary to tion, mitochondrial myopathies, mitochondrial damage (e.g., mitochondrial dysfunction, including reducing frequency calcium accumulation, excitotoxicity, nitric oxide exposure, and severity of seizure activity. hypoxia, etc.), and mitochondrial deregulation. 0428 Metabolic studies on patients with recurrent 0424. A gene defect underlying Friedreich's Ataxia (FA), migraine headaches indicate that deficits in mitochondrial the most common hereditary ataxia, was recently identified activity are commonly associated with this disorder, mani and is designated “frataxin'. In FA, after a period of normal festing as impaired-oxidative phosphorylation and excess development, deficits in coordination develop which progress lactate production. Such deficits are not necessarily due to to paralysis and death, typically between the ages of 30 and genetic defects in mitochondrial DNA. Migraineurs are 40. The tissues affected most severely are the spinal cord, hypersensitive to nitric oxide, an endogenous inhibitor of peripheral nerves, myocardium, and pancreas. Patients typi Cytochrome c Oxidase. In addition, patients with mitochon cally lose motor control and are confined to wheelchairs, and drial cytopathies, e.g. MELAS, often have recurrent are commonly afflicted with heart failure and diabetes. The migraines. In certain embodiments, sirtuin activating com genetic basis for FA involves GAA trinucleotide repeats in an pounds may be used for treating patients with recurrent intron region of the gene encoding frataxin. The presence of migraine headaches, including headaches refractory to ergot these repeats results in reduced transcription and expression compounds or serotonin receptor antagonists. of the gene. Frataxin is involved in regulation of mitochon 0429 Delays in neurological or neuropsychological drial iron content. When cellular frataxin content is subnor development are often found in children with mitochondrial mal, excess iron accumulates in mitochondria, promoting diseases. Development and remodeling of neural connections oxidative damage and consequent mitochondrial degenera requires intensive biosynthetic activity, particularly involving tion and dysfunction. When intermediate numbers of GAA synthesis of neuronal membranes and myelin, both of which repeats are present in the frataxin gene intron, the severe require pyrimidine nucleotides as cofactors. Uridine nucle clinical phenotype of ataxia may not develop. However, these otides are involved inactivation and transfer of Sugars to gly intermediate-length trinucleotide extensions are found in 25 colipids and glycoproteins. Cytidine nucleotides are derived to 30% of patients with non-insulin dependent diabetes mel from uridine nucleotides, and are crucial for synthesis of litus, compared to about 5% of the nondiabetic population. In major membrane phospholipid constituents like phosphati certain embodiments, sirtuin activating compounds may be dylcholine, which receives its choline moiety from cytidine used for treating patients with disorders related to deficien diphosphocholine. In the case of mitochondrial dysfunction cies or defects in frataxin, including Friedreich's Ataxia, (due to either mitochondrial DNA defects or any of the myocardial dysfunction, diabetes mellitus and complications acquired or conditional deficits like exicitoxic or nitric oxide of diabetes like peripheral neuropathy. mediated mitochondrial dysfunction) or other conditions 0425 Muscular dystrophy refers to a family of diseases resulting in impaired pyrimidine synthesis, cell proliferation involving deterioration of neuromuscular structure and func and axonal extension is impaired at crucial stages in devel US 2009/0143376 A1 Jun. 4, 2009

opment of neuronal interconnections and circuits, resulting in low mitochondrial Complex I activity. In certain embodi delayed or arrested development of neuropsychological func ments, sirtuin activating compounds may be useful for treat tions like language, motor, Social, executive function, and ing ALS, for reversing or slowing the progression of clinical cognitive skills. In autism for example, magnetic resonance symptoms. spectroscopy measurements of cerebral phosphate com 0432 Oxygen deficiency results in both direct inhibition pounds indicates that there is global undersynthesis of mem of mitochondrial respiratory chain activity by depriving cells branes and membrane precursors indicated by reduced levels of a terminal electron acceptor for Cytochrome c reoxidation of uridine diphospho-Sugars, and cytidine nucleotide deriva at Complex IV, and indirectly, especially in the nervous sys tives involved in membrane synthesis. Disorders character tem, via secondary post-anoxic excitotoxicity and nitric oxide ized by developmental delay include Rett's Syndrome, per formation. In conditions like cerebral anoxia, angina or sickle vasive developmental delay (or PDD-NOS “pervasive cell anemia crises, tissues are relatively hypoxic. In Such developmental delay not otherwise specified to distinguish it cases, compounds that increase mitochondrial activity pro from specific Subcategories like autism), autism, Asperger's vide protection of affected tissues from deleterious effects of Syndrome, and Attention Deficit/Hyperactivity Disorder hypoxia, attenuate secondary delayed cell death, and accel (ADHD), which is becoming recognized as a delay or lag in erate recovery from hypoxic tissue stress and injury. In certain development of neural circuitry underlying executive func embodiments, sirtuin activating compounds may be useful tions. In certain embodiments, sirtuin activating compounds for preventing delayed cell death (apoptosis in regions like the may be useful for treating patients with neurodevelopmental hippocampus or cortex occurring about 2 to 5 days after an delays (e.g., involving motor, language, executive function, episode of cerebral ischemia) after ischemic or hypoxic insult and cognitive skills), or other delays or arrests of neurological to the brain. and neuropsychological development in the nervous system 0433 Acidosis due to renal dysfunction is often observed and somatic development in non-neural tissues like muscle in patients with mitochondrial disease, whether the underly and endocrine glands. ing respiratory chain dysfunction is congenital or induced by 0430. The two most significant severe neurodegenerative ischemia or cytotoxic agents like cisplatin. Renal tubular diseases associated with aging, Alzheimer's Disease (AD) acidosis often requires administration of exogenous sodium and Parkinson's Disease (PD), both involve mitochondrial bicarbonate to maintain blood and tissue pH. In certain dysfunction in their pathogenesis. Complex I deficiencies in embodiments, sirtuin activating compounds may be useful particular are frequently found not only in the nigrostriatal for treating renal tubular acidosis and other forms of renal neurons that degenerate in Parkinson's disease, but also in dysfunction caused by mitochondrial respiratory chain defi peripheral tissues and cells like muscle and platelets of Par cits. kinson's Disease patients. In Alzheimer's Disease, mitochon 0434. During normal aging, there is a progressive decline drial respiratory chain activity is often depressed, especially in mitochondrial respiratory chain function. Beginning about Complex IV (Cytochrome c Oxidase). Moreover, mitochon age 40, there is an exponential rise in accumulation of mito drial respiratory function altogether is depressed as a conse chondrial DNA defects in humans, and a concurrent decline quence of aging, further amplifying the deleterious sequelae in nuclear-regulated elements of mitochondrial respiratory of additional molecular lesions affecting respiratory chain activity. Many mitochondrial DNA lesions have a selection function. Other factors in addition to primary mitochondrial advantage during mitochondrial turnover, especially in post dysfunction underlie neurodegeneration in AD, PD, and mitotic cells. The proposed mechanism is that mitochondria related disorders. Excitotoxic stimulation and nitric oxide are with a defective respiratory chain produce less oxidative dam implicated in both diseases, factors which both exacerbate age to themselves than do mitochondria with intact functional mitochondrial respiratory chain deficits and whose deleteri respiratory chains (mitochondrial respiration is the primary ous actions are exaggerated on a background of respiratory source of free radicals in the body). Therefore, normally chain dysfunction. Huntington's Disease also involves mito functioning mitochondria accumulate oxidative damage to chondrial dysfunction in affected brain regions, with coop membrane lipids more rapidly than do defective mitochon erative interactions of excitotoxic stimulation and mitochon dria, and are therefore "tagged' for degradation by lysos drial dysfunction contributing to neuronal degeneration. In omes. Since mitochondria within cells have a half life of certain embodiments, sirtuin activating compounds may be about 10 days, a selection advantage can result in rapid useful for treating and attenuating progression of age-related replacement of functional mitochondria with those with neurodegenerative diseases including AD and PD. diminished respiratory activity, especially in slowly dividing 0431 One of the major genetic defects in patients with cells. The net result is that once a mutation in a gene for a Amyotrophic Lateral Sclerosis (ALS or Lou Gehrig's Dis mitochondrial protein that reduces oxidative damage to mito ease) is mutation or deficiency in Copper-Zinc Superoxide chondria occurs, such defective mitochondria will rapidly Dismutase (SOD 1), an antioxidant enzyme. Mitochondria populate the cell, diminishing or eliminating its respiratory both produce and are primary targets for reactive oxygen capabilities. The accumulation of Such cells results in aging species. Inefficient transfer of electrons to oxygen in mito or degenerative disease at the organismal level. This is con chondria is the most significant physiological source of free sistent with the progressive mosaic appearance of cells with radicals in mammalian systems. Deficiencies in antioxidants defective electron transport activity in muscle, with cells or antioxidant enzymes can result in or exacerbate mitochon almost devoid of Cytochromec Oxidase (COX) activity inter drial degeneration. Mice transgenic for mutated SOD1 spersed randomly amidst cells with normal activity, and a develop symptoms and pathology similar to those in human higher incidence of COX-negative cells in biopsies from ALS. The development of the disease in these animals has older Subjects. The organism, during aging, or in a variety of been shown to involve oxidative destruction of mitochondria mitochondrial diseases, is thus faced with a situation in which followed by functional decline of motor neurons and onset of irreplaceable postmitotic cells (e.g. neurons, skeletal and car clinical symptoms. Skeletal muscle from ALS patients has diac muscle) must be preserved and their function maintained US 2009/0143376 A1 Jun. 4, 2009 44 to a significant degree, in the face of an inexorable progressive found in tissues of Down's patients and in late-onset Alzhe decline in mitochondrial respiratory chain function. Neurons imer's Disease. Appropriate Support of mitochondrial func with dysfunctional mitochondria become progressively more tion or compensation for mitochondrial dysfunction therefore sensitive to insults like excitotoxic injury. Mitochondrial fail is useful for protecting againstage-related or chemotherapy ure contributes to most degenerative diseases (especially neu induced menopause or irregularities of menstrual cycling or rodegeneration) that accompany aging. Congenital mito ovulation. In certain embodiments, sirtuin activating com chondrial diseases often involve early-onset pounds may be useful for treating and preventing amenor neurodegeneration similar in fundamental mechanism to dis rhea, irregular ovulation, menopause, or secondary conse orders that occur during aging of people born with normal quences of menopause. mitochondria. In certain embodiments, sirtuin activating 0437. In certain embodiments, sirtuin modulating com compounds may be useful for treating or attenuating cogni pounds may be useful for treatment mitochondrial myopa tive decline and other degenerative consequences of aging. thies. Mitochondrial myopathies range from mild, slowly 0435 Mitochondrial DNA damage is more extensive and progressive weakness of the extraocular muscles to severe, persists longer than nuclear DNA damage in cells Subjected to fatal infantile myopathies and multisystem encephalomyopa oxidative stress or cancer chemotherapy agents like cisplatin thies. Some syndromes have been defined, with some overlap due to both greater Vulnerability and less efficient repair of between them. Established syndromes affecting muscle mitochondrial DNA. Although mitochondrial DNA may be include progressive external ophthalmoplegia, the Kearns more sensitive to damage than nuclear DNA, it is relatively Sayre syndrome (with ophthalmoplegia, pigmentary retin resistant, in Some situations, to mutagenesis by chemical opathy, cardiac conduction defects, cerebellar ataxia, and carcinogens. This is because mitochondria respond to some sensorineural deafness), the MELAS syndrome (mitochon types of mitochondrial DNA damage by destroying their drial encephalomyopathy, lactic acidosis, and stroke-like epi defective genomes rather than attempting to repair them. This sodes), the MERFF syndrome (myoclonic epilepsy and results in global mitochondrial dysfunction for a period after ragged red fibers), limb-girdle distribution weakness, and cytotoxic chemotherapy. Clinical use of chemotherapy agents infantile myopathy (benign or severe and fatal). Muscle like cisplatin, mitomycin, and cytoxan is often accompanied biopsy specimens stained with modified Gomori's trichrome by debilitating “chemotherapy fatigue’, prolonged periods of stain show ragged red fibers due to excessive accumulation of weakness and exercise intolerance which may persist even mitochondria. Biochemical defects in Substrate transport and after recovery from hematologic and gastrointestinal toxici utilization, the Krebs cycle, oxidative phosphorylation, or the ties of such agents. In certain embodiments, sirtuin activating respiratory chain are detectable. Numerous mitochondrial compounds may be useful for treatment and prevention of DNA point mutations and deletions have been described, side effects of cancer chemotherapy related to mitochondrial transmitted in a maternal, nonmendelian inheritance pattern. dysfunction. Mutations in nuclear-encoded mitochondrial enzymes occur. 0436. A crucial function of the ovary is to maintain integ 0438. In certain embodiments, sirtuin activating com rity of the mitochondrial genome in oocytes, since mitochon pounds may be useful for treating patients suffering from dria passed onto a fetus are all derived from those present in toxic damage to mitochondria, such as, toxic damage due to oocytes at the time of conception. Deletions in mitochondrial calcium accumulation, excitotoxicity, nitric oxide exposure, DNA become detectable around the age of menopause, and drug induced toxic damage, or hypoxia. are also associated with abnormal menstrual cycles. Since 0439 A fundamental mechanism of cell injury, especially cells cannot directly detect and respond to defects in mito in excitable tissues, involves excessive calcium entry into chondrial DNA, but can only detect secondary effects that cells, as a result of either leakage through the plasma mem affect the cytoplasm, like impaired respiration, redox status, brane or defects in intracellular calcium handling mecha or deficits in pyrimidine synthesis, such products of mito nisms. Mitochondria are major sites of calcium sequestration, chondrial function participate as a signal for oocyte selection and preferentially utilize energy from the respiratory chain and follicular atresia, ultimately triggering menopause when for taking up calcium rather than for ATP synthesis, which maintenance of mitochondrial genomic fidelity and func results in a downward spiral of mitochondrial failure, since tional activity can no longer be guaranteed. This is analogous calcium uptake into mitochondria results in diminished capa to apoptosis in cells with DNA damage, which undergo an bilities for energy transduction. active process of cellular Suicide when genomic fidelity can 0440 Excessive stimulation of neurons with excitatory no longer beachieved by repair processes. Women with mito amino acids is a common mechanism of cell death or injury in chondrial cytopathies affecting the gonads often undergo pre the central nervous system. Activation of glutamate receptors, mature menopause or display primary cycling abnormalities. especially of the subtype designated NMDA receptors, Cytotoxic cancer chemotherapy often induces premature results in mitochondrial dysfunction, in part through eleva menopause, with a consequent increased risk of osteoporosis. tion of intracellular calcium during excitotoxic stimulation. Chemotherapy-induced amenorrhea is generally due to pri Conversely, deficits in mitochondrial respiration and oxida mary ovarian failure. The incidence of chemotherapy-in tive phosphorylation sensitizes cells to excitotoxic stimuli, duced amenorrhea increases as a function of age in premeno resulting in cell death or injury during exposure to levels of pausal women receiving chemotherapy, pointing toward excitotoxic neurotransmitters or toxins that would be innocu mitochondrial involvement. Inhibitors of mitochondrial res ous to normal cells. piration or protein synthesis inhibit hormone-induced ovula 0441 Nitric oxide (about 1 micromolar) inhibits cyto tion, and furthermore inhibit production of ovarian steroid chrome oxidase (Complex IV) and thereby inhibits mito hormones in response to pituitary gonadotropins. Women chondrial respiration; moreover, prolonged exposure to nitric with Down's syndrome typically undergo menopause prema oxide (NO) irreversibly reduces Complex I activity. Physi turely, and also are subject to early onset of Alzheimer-like ological or pathophysiological concentrations of NOthereby dementia. Low activity of cytochrome oxidase is consistently inhibit pyrimidine biosynthesis. Nitric oxide is implicated in US 2009/0143376 A1 Jun. 4, 2009

a variety of neurodegenerative disorders including inflamma contraction. Athlete refers to an individual who participates in tory and autoimmune diseases of the central nervous system, sports at any leveland who seeks to achieve an improved level and is involved in mediation of excitotoxic and post-hypoxic of strength, speed and endurance in their performance. Such damage to neurons. as, for example, body builders, bicyclists, long distance run 0442. Oxygen is the terminal electron acceptor in the res ners, short distance runners, etc. An athlete may be hard piratory chain. Oxygen deficiency impairs electron transport training, that is, performs sports activities intensely more than chain activity, resulting in diminished pyrimidine synthesis as three days a week or for competition. An athlete may also be well as diminished ATP synthesis via oxidative phosphoryla a fitness enthusiast who seeks to improve general health and tion. Human cells proliferate and retain viability under virtu well-being, improve energy levels, who works out for about ally anaerobic conditions if provided with uridine and 1-2 hours about 3 times a week. Enhanced sports performance pynivate (or a similarly effective agent for oxidizing NADH in manifested by the ability to overcome muscle fatigue, to optimize glycolytic ATP production). ability to maintain activity for longer periods of time, and 0443. In certain embodiments, sirtuin activating com have a more effective workout. pounds may be useful for treating diseases or disorders asso 0448. In the arena of athlete muscle performance, it is ciated with mitochondrial deregulation. desirable to create conditions that permit competition or 0444 Transcription of mitochondrial DNA encoding res training at higher levels of resistance for a prolonged period piratory chain components requires nuclear factors. In neu of time. However, acute and intense anaerobic use of skeletal ronal axons, mitochondria must shuttle back and forth to the muscles often results in impaired athletic performance, with nucleus in order to maintain respiratory chain activity. If losses in force and work output, and increased onset of axonal transport is impaired by hypoxia or by drugs like taxol muscle fatigue, Soreness, and dysfunction. It is now recog which affect microtubule stability, mitochondria distant from nized that even a single exhaustive exercise session, or for that the nucleus undergo loss of cytochrome oxidase activity. matter any acute trauma to the body Such as muscle injury, Accordingly, treatment with a sirtuin activating compound resistance or exhaustive muscle exercise, or elective Surgery, may be useful for promoting nuclear-mitochondrial interac is characterized by perturbed metabolism that affects muscle tions. performance in both short and long term phases. Both muscle 0445 Mitochondria are the primary source of free radicals metabolic/enzymatic activity and gene expression are and reactive oxygen species, due to spillover from the mito affected. For example, disruption of skeletal muscle nitrogen chondrial respiratory chain, especially when defects in one or metabolism as well as depletion of sources of metabolic more respiratory chain components impairs orderly transfer energy occur during extensive muscle activity. Amino acids, of electrons from metabolic intermediates to molecular oxy including branched-chain amino acids, are released from gen. To reduce oxidative damage, cells can compensate by muscles followed by their deamination to elevate serum expressing mitochondrial uncoupling proteins (UCP), of ammonia and local oxidation as muscle fuel Sources, which which several have been identified. UCP-2 is transcribed in augments metabolic acidosis. In addition, there is a decline in response to oxidative-damage, inflammatory cytokines, or catalytic efficiency of muscle contraction events, as well as an excess lipid loads, e.g. fatty liver and steatohepatitis. UCPs alteration of enzymatic activities of nitrogen and energy reduce spillover of reactive oxygen species from mitochon metabolism. Further, protein catabolism is initiated where dria by discharging proton gradients across the mitochondrial rate of protein synthesis is decreased coupled with an increase inner membrane, in effect wasting energy produced by in the degradation of non-contractible protein. These meta metabolism and rendering cells Vulnerable to energy stress as bolic processes are also accompanied by free radical genera a trade-off for reduced oxidative injury. tion which further damages muscle cells. 0449 Recovery from fatigue during acute and extended Muscle Performance exercise requires reversal of metabolic and non-metabolic 0446. In other embodiments, the invention provides meth fatiguing factors. Known factors that participate in human ods for enhancing muscle performance by administering a muscle fatigue. Such as lactate, ammonia, hydrogen ion, etc., therapeutically effective amount of a sirtuin activating com provide an incomplete and unsatisfactory explanation of the pound. For example, sirtuin activating compounds may be fatigue/recovery process, and it is likely that additional useful for improving physical endurance (e.g., ability to per unknown agents participate (Baker et al., J. Appl. Physiol. form a physical task Such as exercise, physical labor, sports 74:2294-2300, 1993; Bazzarre et al., J. Am. Coll. Nutr. activities, etc.), inhibiting or retarding physical fatigues, 11:505-511, 1992: Dohm et al., Fed. Proc. 44:348-352, 1985; enhancing blood oxygen levels, enhancing energy in healthy Edwards In: Biochemistry of Exercise, Proceedings of the individuals, enhance working capacity and endurance, reduc Fifth International Symposium on the Biochemistry of Exer ing muscle fatigue, reducing stress, enhancing cardiac and cise (Kutrgen, Vogel, Poormans, eds.), 1983; MacDougallet cardiovascular function, improving sexual ability, increasing al., Acta Physiol. Scand. 146:403-404, 1992; Walser et al., muscle ATP levels, and/or reducing lactic acid in blood. In Kidney Int. 32:123-128, 1987). Several studies have also certain embodiments, the methods involve administering an analyzed the effects of nutritional supplements and herbal amount of a sirtuin activating compound that increase mito Supplements in enhancing muscle performance. chondrial activity, increase mitochondrial biogenesis, and/or 0450 Aside from muscle performance during endurance increase mitochondrial mass. exercise, free radicals and oxidative stress parameters are 0447 Sports performance refers to the ability of the ath affected in pathophysiological states. A Substantial body of lete's muscles to perform when participating in sports activi data now Suggests that oxidative stress contributes to muscle ties. Enhanced sports performance, strength, speed and wasting or atrophy in pathophysiological states (reviewed in endurance are measured by an increase in muscular contrac Clarkson, P. M. Antioxidants and physical performance. Crit. tion strength, increase in amplitude of muscle contraction, Rev. Food Sci. Nutr. 35: 31-41; 1995; Powers, S. K. Lennon, shortening of muscle reaction time between stimulation and S. L. Analysis of cellular responses to free radicals: Focus on US 2009/0143376 A1 Jun. 4, 2009 46 exercise and skeletal muscle. Proc. Nutr Soc. 58: 1025-1033; bovines, equines, porcines, canines, felines, non-human pri 1999). For example, with respect to muscular disorders where mate, mice, and rats. Cells that may be treated include eukary both muscle endurance and function are compensated, the otic cells, e.g., from a subject described above, or plant cells, role of nitric oxide (NO), has been implicated. In muscular yeast cells and prokaryotic cells, e.g., bacterial cells. For dystrophies, especially those due to defects in proteins that example, modulating compounds may be administered to make up the dystrophin-glycoprotein complex (DGC), the farm animals to improve their ability to withstand farming enzyme that synthesizes NO, nitric oxide synthase (NOS), conditions longer. has been associated. Recent studies of dystrophies related to 0454 Sirtuin-modulating compounds that increase the DGC defects suggest that one mechanism of cellular injury is level and/or activity of a sirtuin protein may also be used to functional ischemia related to alterations in cellular NOS and increase lifespan, stress resistance, and resistance to apopto disruption of a normal protective action of NO. This protec sis in plants. In one embodiment, a compound is applied to tive action is the prevention of localischemia during contrac plants, e.g., on a periodic basis, or to fungi. In another tion-induced increases in sympathetic vasoconstriction. embodiment, plants are genetically modified to produce a Rando (Microsc Res Tech 55(4):223-35, 2001), has shown compound. In another embodiment, plants and fruits are that oxidative injury precedes pathologic changes and that treated with a compound prior to picking and shipping to muscle cells with defects in the DGC have an increased increase resistance to damage during shipping. Plant seeds Susceptibility to oxidant challenges. Excessive lipid peroxi may also be contacted with compounds described herein, e.g., dation due to free radicals has also been shown to be a factor to preserve them. in myopathic diseases such as McArdle's disease (Russo et 0455. In other embodiments, sirtuin-modulating com al., Med. Hypotheses. 39(2): 147-51, 1992). Furthermore, pounds that increase the level and/or activity of a sirtuin mitochondrial dysfunction is a well-known correlate of age protein may be used for modulating lifespan in yeast cells. related muscle wasting (sarcopenia) and free radical damage Situations in which it may be desirable to extend the lifespan has been Suggested, though poorly investigated, as a contrib of yeast cells include any process in which yeast is used, e.g., uting factor (reviewed in Navarro, A.; Lopez-Cepero, J. M.: the making of beer, yogurt, and bakery items, e.g., bread. Use Sanchez del Pino, M. L. Front. Biosci. 6: D2644; 2001). Other of yeast having an extended lifespan can result in using less indications include acute sarcopenia, for example muscle yeast or in having the yeast be active for longer periods of atrophy and/or cachexia associated with burns, bed rest, limb time. Yeast or other mammalian cells used for recombinantly immobilization, or major thoracic, abdominal, and/or ortho producing proteins may also be treated as described herein. pedic Surgery. It is contemplated that the methods of the 0456 Sirtuin-modulating compounds that increase the present invention will also be effective in the treatment of level and/or activity of a sirtuin protein may also be used to muscle related pathological conditions. increase lifespan, stress resistance and resistance to apoptosis 0451. In certain embodiments, the invention provides in insects. In this embodiment, compounds would be applied novel dietary compositions comprising sirtuin modulators, a to useful insects, e.g., bees and other insects that are involved method for their preparation, and a method of using the com in pollination of plants. In a specific embodiment, a com positions for improvement of sports performance. Accord pound would be applied to bees involved in the production of ingly, provided are therapeutic compositions, foods and bev honey. Generally, the methods described herein may be erages that have actions of improving physical endurance applied to any organism, e.g., eukaryote, that may have com and/or inhibiting physical fatigues for those people involved mercial importance. For example, they can be applied to fish in broadly-defined exercises including sports requiring (aquaculture) and birds (e.g., chicken and fowl). endurance and labors requiring repeated muscle exertions. 0457. Higher doses of sirtuin-modulating compounds that Such dietary compositions may additional comprise electro increase the level and/or activity of a sirtuin protein may also lytes, caffeine, Vitamins, carbohydrates, etc. be used as a pesticide by interfering with the regulation of silenced genes and the regulation of apoptosis during devel Other Uses opment. In this embodiment, a compound may be applied to 0452 Sirtuin-modulating compounds that increase the plants using a method known in the art that ensures the com level and/or activity of a sirtuin protein may be used for pound is bio-available to insect larvae, and not to plants. treating or preventing viral infections (such as infections by 0458. At least in view of the link between reproduction influenza, herpes or papilloma virus) or as antifungal agents. and longevity (Longo and Finch, Science, 2002), sirtuin In certain embodiments, sirtuin-modulating compounds that modulating compounds that increase the level and/or activity increase the level and/or activity of a sirtuin protein may be of a sirtuin protein can be applied to affect the reproduction of administered as part of a combination drug therapy with organisms such as insects, animals and microorganisms. another therapeutic agent for the treatment of viral diseases, including, for example, acyclovir, ganciclovir and Zidovu 4. Assays dine. In another embodiment, sirtuin-modulating compounds 0459 Yet other methods contemplated herein include that increase the level and/or activity of a sirtuin protein may screening methods for identifying compounds or agents that be administered as part of a combination drug therapy with modulate sirtuins. An agent may be a nucleic acid. Such as an anotheranti-fungal agent including, for example, topical anti aptamer. Assays may be conducted in a cell based or cell free fungals such as ciclopiroX, clotrimazole, econazole, micona format. For example, an assay may comprise incubating (or Zole, nystatin, oxiconazole, terconazole, and tolnaftate, or contacting) a sirtuin with a test agent under conditions in systemic anti-fungal Such as fluconazole (Diflucan), itracona which a sirtuin can be modulated by an agent known to Zole (Sporanox), ketoconazole (NiZoral), and miconazole modulate the sirtuin, and monitoring or determining the level (Monistat I.V.). of modulation of the sirtuin in the presence of the test agent 0453 Subjects that may be treated as described herein relative to the absence of the test agent. The level of modula include eukaryotes, such as mammals, e.g., humans, Ovines, tion of a sirtuin can be determined by determining its ability US 2009/0143376 A1 Jun. 4, 2009 47 to deacetylate a substrate. Exemplary substrates are acety with the test compound and the substrate may be conducted lated peptides which can be obtained from BIOMOL (Ply for about 10 minutes to 5 hours, preferably for about 1-3 mouth Meeting, Pa.). Preferred substrates include peptides of hours. Since TSA inhibits all class I and class II HDACs, and p53. Such as those comprising an acetylated K382. A particu that certain substrates, e.g., Fluor de Lys, is a poor substrate larly preferred substrate is the Fluor de Lys-SIRT1 (BIO for SIRT2 and even less a substrate for SIRT3-7, such an MOL), i.e., the acetylated peptide Arg-His-Lys-Lys. Other assay may be used to identify modulators of SIRT1 in vivo. Substrates are peptides from human histones H3 and H4 or an acetylated amino acid (see FIG. 5). Substrates may be fluo 5. Pharmaceutical Compositions rogenic. The sirtuin may be SIRT1, Sir2, SIRT3, or a portion 0462. The sirtuin-modulating compounds described thereof. For example, recombinant SIRT1 can be obtained herein may be formulated in a conventional manner using one from BIOMOL. The reaction may be conducted for about 30 or more physiologically acceptable carriers or excipients. For minutes and stopped, e.g., with nicotinamide. The HDAC example, sirtuin-modulating compounds and their physi fluorescent activity assay/drug discovery kit (AK-500, BIO ologically acceptable salts and solvates may be formulated MOL Research Laboratories) may be used to determine the for administration by, for example, injection (e.g. SubQ, IM, level of acetylation. Similar assays are described in Bitterman IP), inhalation or insufflation (either through the mouth or the et al. (2002) J. Biol. Chem. 277:45099. The level of modula nose) or oral, buccal, Sublingual, transdermal, nasal, tion of the sirtuin in an assay may be compared to the level of parenteral or rectal administration. In one embodiment, a modulation of the sirtuin in the presence of one or more sirtuin-modulating compound may be administered locally, (separately or simultaneously) compounds described herein, at the site where the target cells are present, i.e., in a specific which may serve as positive or negative controls. Sirtuins for tissue, organ, or fluid (e.g., blood, cerebrospinal fluid, etc.). use in the assays may be full length sirtuin proteins orportions 0463 Sirtuin-modulating compounds can be formulated thereof. Since it has been shown herein that activating com for a variety of modes of administration, including systemic pounds appear to interact with the N-terminus of SIRT1, and topical or localized administration. Techniques and for proteins for use in the assays include N-terminal portions of mulations generally may be found in Remington's Pharma sirtuins, e.g., about amino acids 1-176 or 1-255 of SIRT1: ceutical Sciences, Meade Publishing Co., Easton, Pa. For about amino acids 1-174 or 1-252 of Sir2. parenteral administration, injection is preferred, including 0460. In one embodiment, a screening assay comprises (i) intramuscular, intravenous, intraperitoneal, and subcutane contacting a sirtuin with a test agent and an acetylated sub ous. For injection, the compounds can beformulated in liquid strate under conditions appropriate for the sirtuin to deacety Solutions, preferably in physiologically compatible buffers late the substrate in the absence of the test agent; and (ii) Such as Hank's solution or Ringer's solution. In addition, the determining the level of acetylation of the substrate, wherein compounds may be formulated in solid form and redissolved a lower level of acetylation of the substrate in the presence of or Suspended immediately prior to use. Lyophilized forms are the test agent relative to the absence of the test agent indicates also included. that the test agent stimulates deacetylation by the sirtuin, 0464 For oral administration, the pharmaceutical compo whereas a higher level of acetylation of the substrate in the sitions may take the form of for example, tablets, lozenges, or presence of the test agent relative to the absence of the test capsules prepared by conventional means with pharmaceuti agent indicates that the test agent inhibits deacetylation by the cally acceptable excipients such as binding agents (e.g., sirtuin. pregelatinised maize starch, polyvinylpyrrolidone or hydrox 0461 Methods for identifying an agent that modulates, ypropyl methylcellulose); fillers (e.g., lactose, microcrystal e.g., stimulates or inhibits, sirtuins in vivo may comprise (i) line cellulose or calcium hydrogen phosphate); lubricants contacting a cell with a test agent and a substrate that is (e.g., magnesium Stearate, talc or silica); disintegrants (e.g., capable of entering a cell in the presence of an inhibitor of potato starch or sodium starch glycolate); or wetting agents class I and class II HDACs under conditions appropriate for (e.g., sodium lauryl Sulphate). The tablets may be coated by the sirtuin to deacetylate the substrate in the absence of the methods well known in the art. Liquid preparations for oral test agent; and (ii) determining the level of acetylation of the administration may take the form of for example, solutions, substrate, wherein a lower level of acetylation of the substrate Syrups or suspensions, or they may be presented as a dry in the presence of the test agent relative to the absence of the product for constitution with water or other suitable vehicle test agent indicates that the test agent stimulates deacetylation before use. Such liquid preparations may be prepared by by the sirtuin, whereas a higher level of acetylation of the conventional means with pharmaceutically acceptable addi Substrate in the presence of the test agent relative to the tives such as suspending agents (e.g., sorbitol syrup, cellulose absence of the test agent indicates that the test agent inhibits derivatives or hydrogenated edible fats); emulsifying agents deacetylation by the sirtuin. A preferred substrate is an acety (e.g., lecithin or acacia); non-aqueous vehicles (e.g., ationd lated peptide, which is also preferably fluorogenic, as further oil, oily esters, ethyl alcohol or fractionated vegetable oils): described herein. The method may further comprise lysing and preservatives (e.g., methyl or propyl-p-hydroxyben the cells to determine the level of acetylation of the substrate. Zoates or sorbic acid). The preparations may also contain Substrates may be added to cells at a concentration ranging buffer salts, flavoring, coloring and sweetening agents as from about 1 uM to about 10 mM, preferably from about 10 appropriate. Preparations for oral administration may be suit uM to 1 mM, even more preferably from about 100 uM to 1 ably formulated to give controlled release of the active com mM, such as about 200 uM. A preferred substrate is an acety pound. lated lysine, e.g., e-acetyl lysine (Fluor de Lys, FdL) or Fluor 0465 For administration by inhalation (e.g., pulmonary de Lys-SIRT1. A preferred inhibitor of class I and class II delivery), sirtuin-modulating compounds may be conve HDACs is trichostatin A (TSA), which may be used at con niently delivered in the form of an aerosol spray presentation centrations ranging from about 0.01 to 100 uM, preferably from pressurized packs or a nebuliser, with the use of a from about 0.1 to 10 uM, such as 1 uM. Incubation of cells Suitable propellant, e.g., dichlorodifluoromethane, trichlorof US 2009/0143376 A1 Jun. 4, 2009 48 luoromethane, dichlorotetrafluoroethane, carbon dioxide or known as "nanocapsules'. In certain embodiments, nanopar other suitable gas. In the case of a pressurized aerosol the ticles are preferred having a diameter from about 50 nm to dosage unit may be determined by providing a valve to deliver about 500 nm, in particular from about 100 nm to about 200 a metered amount. Capsules and cartridges of e.g., gelatin, for . use in an inhaler or insufflator may be formulated containing 0472. Nanoparticles can be prepared by in situ polymer a powder mix of the compound and a Suitable powder base ization of dispersed monomers or by using preformed poly Such as lactose or starch. mers. Since polymers prepared in situ are often not biode 0466 Sirtuin-modulating compounds may be formulated gradable and/or contain toxicological serious byproducts, for parenteral administration by injection, e.g., by bolus nanoparticles from preformed polymers are preferred. Nano injection or continuous infusion. Formulations for injection particles from preformed polymers can be prepared by dif may be presented in unit dosage form, e.g., in ampoules or in ferent techniques, e.g., by emulsion evaporation, Solvent dis multi-dose containers, with an added preservative. The com placement, salting-out, mechanical grinding, positions may take such forms as Suspensions, solutions or microprecipitation, and by emulsification diffusion. emulsions in oily or aqueous vehicles, and may contain for 0473 With the methods described above, nanoparticles mulatory agents such as Suspending, stabilizing and/or dis can be formed with various types of polymers. For use in the persing agents. Alternatively, the active ingredient may be in method of the present invention, nanoparticles made from powder form for constitution with a suitable vehicle, e.g., biocompatible polymers are preferred. The term “biocompat sterile pyrogen-free water, before use. ible' refers to material that after introduction into a biological 0467 Sirtuin-modulating compounds may also be formu environment has no serious effects to the biological environ lated in rectal compositions such as Suppositories or retention ment. From biocompatible polymers those polymers are enemas, e.g., containing conventional Suppository bases Such especially preferred which are also biodegradable. The term as cocoa butter or other glycerides. “biodegradable' refers to material that after introduction into 0468. In addition to the formulations described previously, a biological environment is enzymatically or chemically sirtuin-modulating compounds may also be formulated as a degraded into Smaller molecules, which can be eliminated depot preparation. Such long acting formulations may be Subsequently. Examples are polyesters from hydroxycar administered by implantation (for example Subcutaneously or boxylic acids such as poly(lactic acid) (PLA), poly(glycolic intramuscularly) or by intramuscular injection. Thus, for acid) (PGA), polycaprolactone (PCL), copolymers of lactic example, sirtuin-modulating compounds may be formulated acid and glycolic acid (PLGA), copolymers of lactic acid and with suitable polymeric or hydrophobic materials (for caprolactone, polyepsilon caprolactone, polyhyroxybutyric example as an emulsion in an acceptable oil) or ion exchange acid and poly(ortho)esters, polyurethanes, polyanhydrides, resins, or as sparingly soluble derivatives, for example, as a polyacetals, polydihydropyrians, polycyanoacrylates, natural sparingly soluble salt. Controlled release formula also polymers such as alginate and other polysaccharides includ includes patches. ing dextran and cellulose, collagen and albumin. 0469. In certain embodiments, the compounds described 0474 Suitable surface modifiers can preferably be herein can be formulated for delivery to the central nervous selected from known organic and inorganic pharmaceutical system (CNS) (reviewed in Begley, Pharmacology & Thera excipients. Such excipients include various polymers, low peutics 104: 29-45 (2004)). Conventional approaches for molecular weight oligomers, natural products and Surfac drug delivery to the CNS include: neurosurgical strategies tants. Preferred surface modifiers include nonionic and ionic (e.g., intracerebral injection or intracerebroVentricular influ Surfactants. Representative examples of Surface modifiers sion); molecular manipulation of the agent (e.g., production include gelatin, casein, lecithin (phosphatides), gum acacia, of a chimeric fusion protein that comprises a transport peptide cholesterol, tragacanth, Stearic acid, benzalkonium chloride, that has an affinity for an endothelial cell surface molecule in calcium Stearate, glycerol monostearate, cetostearyl alcohol, combination with an agent that is itself incapable of crossing cetomacrogol emulsifying wax, Sorbitan esters, polyoxyeth the BBB) in an attempt to exploit one of the endogenous ylene alkyl ethers, e.g., macrogolethers such ascetomacrogol transport pathways of the BBB. pharmacological strategies 1000, polyoxyethylene castor oil derivatives, polyoxyethyl designed to increase the lipid solubility of an agent (e.g., ene Sorbitan fatty acid esters, e.g., the commercially available conjugation of water-soluble agents to lipid or cholesterol TweensTM, polyethylene glycols, polyoxyethylene stearates, carriers); and the transitory disruption of the integrity of the colloidal silicon dioxide, phosphates, Sodium dodecylsulfate, BBB by hyperosmotic disruption (resulting from the infusion carboxymethylcellulose calcium, carboxymethylcellulose of a mannitol solution into the carotid artery or the use of a sodium, methylcellulose, hydroxyethylcellulose, hydroxy biologically active agent Such as an angiotensin peptide). propylcellulose, hydroxypropylmethylcellulose phthalate, 0470. One possibility to achieve sustained release kinetics noncrystalline cellulose, magnesium aluminum silicate, tri is embedding or encapsulating the active compound into ethanolamine, polyvinyl alcohol, and polyvinylpyrrolidone nanoparticles. Nanoparticles can be administrated as powder, (PVP). Most of these surface modifiers are known pharma as a powder mixture with added excipients or as Suspensions. ceutical excipients and are described in detail in the Hand Colloidal suspensions of nanoparticles can easily be admin book of Pharmaceutical Excipients, published jointly by the istrated through a cannula with Small diameter. American Pharmaceutical Association and The Pharmaceu 0471 Nanoparticles are particles with a diameter from tical Society of Great Britain, the Pharmaceutical Press, 1986. about 5 nm to up to about 1000 nm. The term “nanoparticles' 0475. Further description on preparing nanoparticles can as it is used hereinafter refers to particles formed by a poly be found, for example, in U.S. Pat. No. 6,264,922, the con meric matrix in which the active compound is dispersed, also tents of which are incorporated herein by reference. known as "nanospheres', and also refers to nanoparticles 0476 Liposomes are a further drug delivery system which which are composed of a core containing the active com is easily injectable. Accordingly, in the method of invention pound which is surrounded by a polymeric membrane, also the active compounds can also be administered in the form of US 2009/0143376 A1 Jun. 4, 2009 49 a liposome delivery system. Liposomes are well-known by a sensitive endosomal membrane-disruptive synthetic poly person skilled in the art. Liposomes can be formed from a mers, such as PPAA or PEAA. Certain poly(alkylacrylic variety of phospholipids, such as cholesterol, Stearylamine of acids) have been shown to disrupt endosomal membranes phosphatidylcholines. Liposomes being usable for the while leaving the outer cell surface membrane intact (Stayton method of invention encompass all types of liposomes includ et al. (2000) J. Controll. Release 65:203-220; Murthy et al. ing, but not limited to, Small unilamellar vesicles, large unila (1999) J. Controll. Release 61:137-143: WO 99/34831), mellar vesicles and multilamellar vesicles. thereby increasing cellular bioavailability and functioning as 0477 Liposomes are used for a variety of therapeutic pur a targeting factor. However, PPAA reduces binding of serum poses, and in particular, for carrying therapeutic agents to complement to complexes in which it is incorporated, thus target cells. Advantageously, liposome-drug formulations functioning as a shielding moiety. offer the potential of improved drug-delivery properties, 0482 Another way to produce a formulation, particularly which include, for example, controlled drug release. An a solution, of a sirtuin modulator Such as resveratrol or a extended circulation time is often needed for liposomes to derivative thereof, is through the use of cyclodextrin. By reach a target region, cell or site. In particular, this is neces cyclodextrin is meant Cl-, 3-, or Y-cyclodextrin. Cyclodextrins sary where the target region, cell or site is not located near the are described in detail in Pitha et al., U.S. Pat. No. 4,727,064, site of administration. For example, when liposomes are which is incorporated herein by reference. Cyclodextrins are administered systemically, it is desirable to coat the lipo cyclic oligomers of glucose; these compounds form inclusion Somes with a hydrophilic agent, for example, a coating of complexes with any drug whose molecule can fit into the hydrophilic polymer chains such as polyethylene glycol lipophile-seeking cavities of the cyclodextrin molecule. (PEG) to extend the blood circulation lifetime of the lipo 0483 The cyclodextrin of the compositions according to Somes. Such Surface-modified liposomes are commonly the invention may be Cl-, 3-, or Y-cyclodextrin. C-cyclodextrin referred to as “long circulating or “sterically stabilized contains six glucopyranose units; B-cyclodextrin contains liposomes. seven glucopyranose units; and Y-cyclodextrin contains eight 0478. One surface modification to a liposome is the attach glucopyranose units. The molecule is believed to form a trun ment of PEG chains, typically having a molecular weight cated cone having a core opening of 4.7-5.3 angstroms, 6.0- from about 1000 daltons (Da) to about 5000 Da, and to about 6.5 angstroms, and 7.5-8.3 angstroms in C-, 3-, or Y-cyclo 5 mole percent (%) of the lipids making up the liposomes dextrin respectively. The composition according to the (see, for example, Stealth Liposomes, CRC Press, Lasic, D. invention may comprise a mixture of two or more of the Cl-, and Martin, F., eds. Boca Raton, Fla., (1995)), and the cited B-, or Y-cyclodextrins. Typically, however, the composition references therein. The pharmacokinetics exhibited by such according to the invention will comprise only one of the Cl-, liposomes are characterized by a dose-independent reduction B-, or Y-cyclodextrins. in uptake of liposomes by the liver and spleen via the mono 0484 Most preferred cyclodextrins in the compositions nuclear phagocyte system (MPS), and significantly pro according to the invention are amorphous cyclodextrin com longed blood circulation time, as compared to non-surface pounds. By amorphous cyclodextrin is meant non-crystalline modified liposomes, which tend to be rapidly removed from mixtures of cyclodextrins wherein the mixture is prepared the blood and accumulated in the liver and spleen. from Cl-, 3-, or Y-cyclodextrin. In general, the amorphous 0479. In certain embodiments, the complex is shielded to cyclodextrin is prepared by non-selective alkylation of the increase the circulatory half-life of the complex or shielded to desired cyclodextrin species. Suitable alkylation agents for increase the resistance of nucleic acid to degradation, for this purpose include but are not limited to propylene oxide, example degradation by nucleases. glycidol, iodoacetamide, chloroacetate, and 2-diethylamino 0480. As used herein, the term "shielding, and its cog ethlychloride. Reactions are carried out to yield mixtures nates such as "shielded, refers to the ability of "shielding containing a plurality of components thereby preventing crys moieties' to reduce the non-specific interaction of the com tallization of the cyclodextrin. Various alkylated cyclodex plexes described herein with serum complement or with other trins can be made and of course will vary, depending upon the species present in serum in vitro or in vivo. Shielding moieties starting species of cyclodextrin and the alkylating agent used. may decrease the complex interaction with orbinding to these Among the amorphous cyclodextrins suitable for composi species through one or more mechanisms, including, for tions according to the invention are hydroxypropyl, hydroxy example, non-specific steric or non-specific electronic inter ethyl, glucosyl, maltosyland maltotriosyl derivatives off-cy actions. Examples of Such interactions include non-specific clodextrin, carboxyamidomethyl-3-cyclodextrin, electrostatic interactions, charge interactions, Van der Waals carboxymethyl-3-cyclodextrin, hydroxypropyl-3-cyclodex interactions, steric-hindrance and the like. For a moiety to act trin and diethylamino-3-cyclodextrin. as a shielding moiety, the mechanism or mechanisms by 0485 One example of resveratrol dissolved in the pres which it may reduce interaction with, association with or ence of a cyclodextrin is provided in Marier et al., J. Phar binding to the serum complement or other species does not macol. Exp. Therap. 302:369-373 (2002), the contents of have to be identified. One can determine whethera moiety can which are incorporated herein by reference, where a 6 mg/mL act as a shielding moiety by determining whether or to what solution of resveratrol was prepared using 0.9% saline con extent a complex binds serum species. taining 20% hydroxylpropyl-f-cyclodextrin. 0481. It should be noted that "shielding moieties' can be 0486. As mentioned above, the compositions of matter of multifunctional. For example, a shielding moiety may also the invention comprise an aqueous preparation of preferably function as, for example, a targeting factor. A shielding moi Substituted amorphous cyclodextrin and one or more sirtuin ety may also be referred to as multifunctional with respect to modulators. The relative amounts of sirtuin modulators and the mechanism(s) by which it shields the complex. While not cyclodextrin will vary depending upon the relative amount of wishing to be limited by proposed mechanism or theory, each of the sirtuin modulators and the effect of the cyclodex examples of Such a multifunctional shielding moiety are pH trin on the compound. In general, the ratio of the weight of US 2009/0143376 A1 Jun. 4, 2009 50 compound of the sirtuin modulators to the weight of cyclo machinery. The granules can be combined with a variety of dextrin compound will be in a range between 1:1 and 1:100. carriers including low density, high moldability saccharides, A weight to weight ratio in a range of 1:5 to 1:50 and more low moldability saccharides, polyol combinations, and then preferably in a range of 1:10 to 1:20 of the compound selected directly compressed into a tablet that exhibits an improved from sirtuin modulators to cyclodextrin are believed to be the dissolution and disintegration profile. most effective for increased circulating availability of the sirtuin modulator. 0492. The tablets according to the present invention typi 0487. Importantly, if the aqueous solution comprising the cally have a hardness of about 2 to about 6 Strong-Cobb units sirtuin modulators and a cyclodextrin is to be administered (scu). Tablets within this hardness range disintegrate or dis parenterally, especially via the intravenous route, a cyclodex solve rapidly when chewed. Additionally, the tablets rapidly trin will be substantially free of pyrogenic contaminants. disentegrate in water. On average, a typical 1.1 to 1.5 gram Various forms of cyclodextrin, such as forms of amorphous tablet disintegrates in 1-3 minutes without stirring. This rapid cyclodextrin, may be purchased from a number of vendors disintegration facilitates delivery of the active material. including Sigma-Aldrich, Inc. (St. Louis, Mo., USA). A 0493. The granules used to make the tablets can be, for method for the production of hydroxypropyl-3-cyclodextrin example, mixtures of low density alkali earth metal salts or is disclosed in Pitha et al., U.S. Pat. No. 4,727,064 which is carbohydrates. For example, a mixture of alkali earth metal incorporated herein by reference. salts includes a combination of calcium carbonate and mag 0488. Additional description of the use of cyclodextrin for nesium hydroxide. Similarly, a fast melt tablet can be pre solubilizing compounds can be found in US 2005/0026849, pared according to the methods of the present invention that the contents of which are incorporated herein by reference. incorporates the use of A) spray dried extra light calcium 0489 Rapidly disintegrating or dissolving dosage forms carbonate/maltodextrin, B) magnesium hydroxide and C) a are useful for the rapid absorption, particularly buccal and eutectic polyol combination including Sorbitol Instant, Xyli Sublingual absorption, of pharmaceutically active agents. tol and mannitol. These materials have been combined to Fast melt dosage forms are beneficial to patients, such as aged produce a low density tablet that dissolves very readily and and pediatric patients, who have difficulty in Swallowing promotes the fast disintegration of the active ingredient. typical Solid dosage forms, such as caplets and tablets. Addi Additionally, the pre-compacted and spray dried granules can tionally, fast melt dosage forms circumvent drawbacks asso be combined in the same tablet. ciated with, for example, chewable dosage forms, wherein the length of time an active agent remains in a patient's mouth 0494 For fast melt tablet preparation, a sirtuin modulator plays an important role in determining the amount of taste useful in the present invention can be in a form such as Solid, masking and the extent to which a patient may object to throat particulate, granular, crystalline, oily or Solution. The sirtuin grittiness of the active agent. modulator for use in the present invention may be a spray 0490. To overcome such problems manufacturers have dried product or an adsorbate that has been pre-compacted to developed a number of fast melt solid dose oral formulations. a harder granular form that reduces the medicament taste. A These are available from manufacturers including Cima pharmaceutical active ingredient for use in the present inven Labs, FuisZ Technologies Ltd., Prographarm, R. P. Scherer, tion may be spray dried with a carrier that prevents the active Yamanouchi-Shaklee, and McNeil-PPC, Inc. All of these ingredient from being easily extracted from the tablet when manufacturers market different types of rapidly dissolving chewed. Solid oral dosage forms. See e.g., patents and publications by 0495. In addition to being directly added to the tablets of Cima Labs such as U.S. Pat. No. 5,607,697, U.S. Pat. No. the present invention, the medicament drug itself can be pro 5,503,846, U.S. Pat. No. 5,223,264, U.S. Pat. No. 5,401,513, cessed by the pre-compaction process to achieve an increased U.S. Pat. No. 5,219,574, and U.S. Pat. No. 5,178,878, WO density prior to being incorporated into the formulation. 98/46215, WO 98/14179; patents to Fuisz Technologies, now part of BioVail, such as U.S. Pat. No. 5,871,781, U.S. Pat. No. 0496 The pre-compaction process used in the present 5,869,098, U.S. Pat. No. 5,866,163, U.S. Pat. No. 5,851,553, invention can be used to deliver poorly soluble pharmaceuti U.S. Pat. No. 5,622,719, U.S. Pat. No. 5,567,439, and U.S. cal materials so as to improve the release of such pharmaceu Pat. No. 5,587,172; U.S. Pat. No. 5,464,632 to Prographarm: tical materials over traditional dosage forms. This could allow patents to R. P. Scherer such as U.S. Pat. No. 4,642,903, U.S. for the use of lower dosage levels to deliver equivalent bio Pat. No. 5,188,825, U.S. Pat. No. 5,631,023 and U.S. Pat. No. available levels of drug and thereby lower toxicity levels of 5,827.541; patents to Yamanouchi-Shaklee such as U.S. Pat. both currently marketed drug and new chemical entities. No. 5,576,014 and U.S. Pat. No. 5,446.464; patents to Janssen Poorly soluble pharmaceutical materials can be used in the such as U.S. Pat. No. 5,807,576, U.S. Pat. No. 5,635,210, U.S. form of nanoparticles, which are nanometer-sized particles. Pat. No. 5,595,761, U.S. Pat. No. 5,587,180 and U.S. Pat. No. 0497. In addition to the active ingredient and the granules 5,776,491; U.S. Pat. Nos. 5,639,475 and 5,709,886 to Eurand prepared from low density alkali earth metal salts and/or America, Inc.; U.S. Pat. Nos. 5,807,578 and 5,807,577 to water soluble carbohydrates, the fast melt tablets can be for L.A.B. Pharmaceutical Research; patents to Schering Corpo mulated using conventional carriers or excipients and well ration such as U.S. Pat. Nos. 5,112,616 and 5,073,374; U.S. established pharmaceutical techniques. Conventional carri Pat. No. 4,616,047 to Laboratoire L. LaFon; U.S. Pat. No. ers or excipients include, but are not limited to, diluents, 5,501,861 to Takeda Chemicals Inc., Ltd.; and U.S. Pat. No. binders, adhesives (i.e., cellulose derivatives and acrylic 6,316,029 to Elan. derivatives), lubricants (i.e., magnesium or calcium Stearate, 0491. In one example of fast melt tablet preparation, gran Vegetable oils, polyethylene glycols, talc, sodium lauryl Sul ules for fast melt tablets made by either the spray drying or phate, polyoxyethylene monostearate), disintegrants, colo pre-compacting processes are mixed with excipients and rants, flavorings, preservatives, Sweeteners and miscella compressed into tablets using conventional tablet making neous materials such as buffers and adsorbents. US 2009/0143376 A1 Jun. 4, 2009

0498. Additional description of the preparation of fast lation for use in conjunction with the present method contains melt tablets can be found, for example, in U.S. Pat. No. propylene glycol mixed with a hydrophilic petrolatum Such as 5,939,091, the contents of which are incorporated herein by that which may be obtained under the trademark Aquaphor(R) reference. from Beiersdorf, Inc. (Norwalk, Conn.). 0499 Pharmaceutical compositions (including cosmetic 0504 Sirtuin-modulating compounds may be incorpo preparations) may comprise from about 0.00001 to 100% rated into creams, which generally are viscous liquid or semi such as from 0.001 to 10% or from 0.1% to 5% by weight of solid emulsions, either oil-in-water or water-in-oil. Cream one or more sirtuin-modulating compounds described herein. bases are water-washable, and contain an oil phase, an emul 0500. In one embodiment, a sirtuin-modulating com sifier and an aqueous phase. The oil phase is generally com pound described herein, is incorporated into a topical formu prised of petrolatum and a fatty alcohol Such as cetyl or lation containing a topical carrier that is generally Suited to Stearyl alcohol; the aqueous phase usually, although not nec topical drug administration and comprising any Such material essarily, exceeds the oil phase in Volume, and generally con known in the art. The topical carrier may be selected so as to tains a humectant. The emulsifier in a cream formulation, as provide the composition in the desired form, e.g., as an oint explained in Remington's, Supra, is generally a nonionic, ment, lotion, cream, microemulsion, gel, oil, Solution, or the anionic, cationic or amphoteric Surfactant. like, and may be comprised of a material of either naturally 0505 Sirtuin-modulating compounds may be incorpo occurring or synthetic origin. It is preferable that the selected rated into microemulsions, which generally are thermody carrier not adversely affect the active agent or other compo namically stable, isotropically clear dispersions of two nents of the topical formulation. Examples of suitable topical immiscible liquids, such as oil and water, Stabilized by an carriers for use herein include water, alcohols and other non interfacial film of surfactant molecules (Encyclopedia of toxic organic solvents, glycerin, mineral oil, silicone, petro Pharmaceutical Technology (New York: Marcel Dekker, leum jelly, lanolin, fatty acids, vegetable oils, parabens, 1992), volume 9). For the preparation of microemulsions, waxes, and the like. Surfactant (emulsifier), co-surfactant (co-emulsifier), an oil 0501 Formulations may be colorless, odorless ointments, phase and a water phase are necessary. Suitable surfactants lotions, creams, microemulsions and gels. include any Surfactants that are useful in the preparation of 0502 Sirtuin-modulating compounds may be incorpo emulsions, e.g., emulsifiers that are typically used in the rated into ointments, which generally are semisolid prepara preparation of creams. The co-surfactant (or “co-emulsifer') tions which are typically based on petrolatum or other petro is generally selected from the group of polyglycerol deriva leum derivatives. The specific ointment base to be used, as tives, glycerol derivatives and fatty alcohols. Preferred emul will be appreciated by those skilled in the art, is one that will sifier/co-emulsifier combinations are generally although not provide for optimum drug delivery, and, preferably, will pro necessarily selected from the group consisting of glyceryl vide for other desired characteristics as well, e.g., emolliency monostearate and polyoxyethylene Stearate; polyethylene or the like. As with other carriers or vehicles, anointment base glycol and ethylene glycol palmitostearate; and caprilic and should be inert, stable, nonirritating and nonsensitizing. As capric triglycerides and oleoyl macrogolglycerides. The explained in Remington's (Supra) ointment bases may be water phase includes not only water but also, typically, buff grouped in four classes: oleaginous bases; emulsifiable bases; ers, glucose, propylene glycol, polyethylene glycols, prefer emulsion bases; and water-soluble bases. Oleaginous oint ably lower molecular weight polyethylene glycols (e.g., PEG ment bases include, for example, vegetable oils, fats obtained 300 and PEG 400), and/or glycerol, and the like, while the oil from animals, and semisolid hydrocarbons obtained from phase will generally comprise, for example, fatty acid esters, petroleum. Emulsifiable ointment bases, also known as absor modified vegetable oils, silicone oils, mixtures of mono- di bent ointment bases, contain little or no water and include, for and triglycerides, mono- and di-esters of PEG (e.g., oleoyl example, hydroxyStearin Sulfate, anhydrous lanolin and macrogol glycerides), etc. hydrophilic petrolatum. Emulsion ointment bases are either 0506 Sirtuin-modulating compounds may be incorpo water-in-oil (W/O) emulsions or oil-in-water (O/W) emul rated into gel formulations, which generally are semisolid sions, and include, for example, cetyl alcohol, glyceryl systems consisting of either Suspensions made up of Small monostearate, lanolin and Stearic acid. Exemplary water inorganic particles (two-phase systems) or large organic mol soluble ointment bases are prepared from polyethylene gly ecules distributed substantially uniformly throughout a car cols (PEGs) of varying molecular weight; again, reference rier liquid (single phase gels). Single phase gels can be made, may be had to Remington's, Supra, for further information. for example, by combining the active agent, a carrier liquid 0503 Sirtuin-modulating compounds may be incorpo and a suitable gelling agent Such as tragacanth (at 2 to 5%), rated into lotions, which generally are preparations to be sodium alginate (at 2-10%), gelatin (at 2-15%), methylcellu applied to the skin Surface without friction, and are typically lose (at 3-5%), sodium carboxymethylcellulose (at 2-5%), liquid or semiliquid preparations in which solid particles, carbomer (at 0.3-5%) or polyvinyl alcohol (at 10-20%) including the active agent, are present in a water or alcohol together and mixing until a characteristic semisolid product is base. Lotions are usually Suspensions of Solids, and may produced. Other Suitable gelling agents include methylhy comprise a liquid oily emulsion of the oil-in-water type. droxycellulose, polyoxyethylene-polyoxypropylene, Lotions are preferred formulations for treating large body hydroxyethylcellulose and gelatin. Although gels commonly areas, because of the ease of applying a more fluid composi employ aqueous carrier liquid, alcohols and oils can be used tion. It is generally necessary that the insoluble matter in a as the carrier liquid as well. lotion be finely divided. Lotions will typically contain sus 0507 Various additives, known to those skilled in the art, pending agents to produce better dispersions as well as com may be included in formulations, e.g., topical formulations. pounds useful for localizing and holding the active agent in Examples of additives include, but are not limited to, solubi contact with the skin, e.g., methylcellulose, sodium car lizers, skin permeation enhancers, opacifiers, preservatives boxymethylcellulose, or the like. An exemplary lotion formu (e.g., anti-oxidants), gelling agents, buffering agents, Surfac US 2009/0143376 A1 Jun. 4, 2009 52 tants (particularly nonionic and amphoteric Surfactants), more preferably in the range of approximately 0.5 wt.% to 15 emulsifiers, emollients, thickening agents, stabilizers, wt.% of the formulation, and most preferably in the range of humectants, colorants, fragrance, and the like. Inclusion of approximately 1.0 wt.% to 10 wt.% of the formulation. solubilizers and/or skin permeation enhancers is particularly 0513 Topical skin treatment compositions can be pack preferred, along with emulsifiers, emollients and preserva aged in a Suitable container to Suit its viscosity and intended tives. An optimum topical formulation comprises approxi use by the consumer. For example, a lotion or cream can be mately: 2 wt.% to 60 wt.%, preferably 2 wt.% to 50 wt.%, packaged in a bottle or a roll-ball applicator, or a propellant solubilizer and/or skin permeation enhancer; 2 wt.% to 50 wt. driven aerosol device or a container fitted with a pump Suit %, preferably 2 wt.% to 20 wt.%, emulsifiers; 2 wt.% to 20 able for finger operation. When the composition is a cream, it wt.% emollient; and 0.01 to 0.2 wt.% preservative, with the can simply be stored in a non-deformable bottle or Squeeze active agent and carrier (e.g., water) making of the remainder container, Such as a tube or a lidded jar. The composition may of the formulation. also be included in capsules such as those described in U.S. 0508 A skin permeation enhancer serves to facilitate pas Pat. No. 5,063,507. Accordingly, also provided are closed sage of therapeutic levels of active agent to pass through a containers containing a cosmetically acceptable composition reasonably sized area of unbroken skin. Suitable enhancers as herein defined. are well known in the art and include, for example: lower 0514. In an alternative embodiment, a pharmaceutical for alkanols such as methanol ethanol and 2-propanol; alkyl mulation is provided for oral or parenteral administration, in methyl sulfoxides such as dimethylsulfoxide (DMSO), which case the formulation may comprises a modulating decylmethylsulfoxide (CoMSO) and tetradecylmethyl sulf compound-containing microemulsion as described above, boxide:pyrrolidones such as 2-pyrrolidone, N-methyl-2-pyr but may contain alternative pharmaceutically acceptable car rolidone and N-(-hydroxyethyl)pyrrolidone; urea; N,N-di riers, vehicles, additives, etc. particularly Suited to oral or ethyl-m-toluamide; C-C alkanediols; miscellaneous parenteral drug administration. Alternatively, a modulating solvents such as dimethyl formamide (DMF), N,N-dimethy compound-containing microemulsion may be administered lacetamide (DMA) and tetrahydrofurfuryl alcohol; and the orally or parenterally substantially as described above, with 1-substituted azacycloheptan-2-ones, particularly 1-n-dode out modification. cylcyclazacycloheptan-2-one (laurocapram; available under 0515 Phospholipids complexes, e.g., resveratrol-phos the trademark AZone(R) from Whitby Research Incorporated, pholipid complexes, and their preparation are described in Richmond, Va.). U.S. Patent Application Publication No. 2004/116386. Meth 05.09 Examples of solubilizers include, but are not limited ods for stabilizing active components using polyol/polymer to, the following: hydrophilic ethers such as diethylene glycol microcapsules, and their preparation are described in monoethyl ether (ethoxydiglycol, available commercially as US20040108608. Processes for dissolving lipophilic com Transcutol R) and diethylene glycol monoethyl ether oleate pounds in aqueous Solution with amphiphilic block copoly (available commercially as SoftcutolR); polyethylene castor mers are described in WO 04/035O13. oil derivatives such as polyoxy 35 castor oil, polyoxy 40 0516 Conditions of the eye can be treated or prevented by, hydrogenated castor oil, etc.; polyethylene glycol, particu e.g., systemic, topical, intraocular injection of a sirtuin larly lower molecular weight polyethylene glycols such as modulating compound, or by insertion of a Sustained release PEG 300 and PEG 400, and polyethylene glycol derivatives device that releases a sirtuin-modulating compound. A sir such as PEG-8 caprylic/capric glycerides (available commer tuin-modulating compound that increases or decreases the cially as Labrasol(R); alkyl methylsulfoxides such as DMSO; level and/or activity of a sirtuin protein may be delivered in a pyrrolidones Such as 2-pyrrolidone and N-methyl-2-pyrroli pharmaceutically acceptable ophthalmic vehicle. Such that done; and DMA. Many solubilizers can also act as absorption the compound is maintained in contact with the ocular Surface enhancers. A single solubilizer may be incorporated into the for a sufficient time period to allow the compound to penetrate formulation, or a mixture of solubilizers may be incorporated the corneal and internal regions of the eye, as for example the therein. anterior chamber, posterior chamber, vitreous body, aqueous 0510 Suitable emulsifiers and co-emulsifiers include, humor, vitreous humor, cornea, iris/ciliary, lens, choroid/ without limitation, those emulsifiers and co-emulsifiers retina and Sclera. The pharmaceutically-acceptable oph described with respect to microemulsion formulations. Emol thalmic vehicle may, for example, be an ointment, vegetable lients include, for example, propylene glycol, glycerol, iso oil or an encapsulating material. Alternatively, the com propyl myristate, polypropylene glycol-2 (PPG-2) myristyl pounds of the invention may be injected directly into the ether propionate, and the like. vitreous and aqueous humour. In a further alternative, the 0511. Other active agents may also be included in formu compounds may be administered systemically, such as by lations, e.g., other anti-inflammatory agents, analgesics, anti intravenous infusion or injection, for treatment of the eye. microbial agents, antifungal agents, antibiotics, vitamins, 0517 Sirtuin-modulating compounds described herein antioxidants, and Sunblock agents commonly found in Sun may be stored in oxygen free environment according to meth screen formulations including, but not limited to, anthra ods in the art. For example, resveratrol or analog thereof can nilates, benzophenones (particularly benzophenone-3), cam be prepared in an airtight capsule for oral administration, Such phor derivatives, cinnamates (e.g., octyl methoxycinnamate), as Capsugel from Pfizer, Inc. dibenzoyl methanes (e.g., butyl methoxydibenzoyl methane), 0518 Cells, e.g., treated ex vivo with a sirtuin-modulating p-aminobenzoic acid (PABA) and derivatives thereof, and compound, can be administered according to methods for salicylates (e.g., octyl salicylate). administering a graft to a subject, which may be accompa 0512. In certain topical formulations, the active agent is nied, e.g., by administration of an immunosuppressant drug, present in an amount in the range of approximately 0.25 wt.% e.g., cyclosporin A. For general principles in medicinal for to 75 wt.% of the formulation, preferably in the range of mulation, the reader is referred to Cell Therapy: Stem Cell approximately 0.25 wt.% to 30 wt.% of the formulation, Transplantation, Gene Therapy, and Cellular Immuno US 2009/0143376 A1 Jun. 4, 2009 therapy, by G. Morstyn & W. Sheridan eds, Cambridge Uni Hames & S. J. Higgins eds. 1984); Transcription And Trans versity Press, 1996; and Hematopoietic StemCell Therapy, E. lation (B. D. Hames & S. J. Higgins eds. 1984); Culture Of D. Ball, J. Lister & P. Law, Churchill Livingstone, 2000. Animal Cells (R. I. Freshney, Alan R. Liss, Inc., 1987); 0519 Toxicity and therapeutic efficacy of sirtuin-modu Immobilized Cells And Enzymes (IRL Press, 1986); B. Per lating compounds can be determined by Standard pharmaceu bal, A Practical Guide To Molecular Cloning (1984); the tical procedures in cell cultures or experimental animals. The treatise, Methods. In Enzymology (Academic Press, Inc., LDso is the dose lethal to 50% of the population. The EDs is the dose therapeutically effective in 50% of the population. N.Y.); Gene Transfer Vectors For Mammalian Cells (J. H. The dose ratio between toxic and therapeutic effects (LDs/ Miller and M. P. Calos eds., 1987, Cold Spring Harbor Labo EDso) is the therapeutic index. Sirtuin-modulating com ratory); Methods In Enzymology, Vols. 154 and 155 (Wu et al. pounds that exhibit large therapeutic indexes are preferred. eds.), Immunochemical Methods In Cell And Molecular While sirtuin-modulating compounds that exhibit toxic side Biology (Mayer and Walker, eds., Academic Press, London, effects may be used, care should be taken to design a delivery 1987); Handbook Of Experimental Immunology, Volumes system that targets such compounds to the site of affected I-IV (D. M. Weir and C. C. Blackwell, eds., 1986); Manipu tissue in order to minimize potential damage to uninfected lating the Mouse Embryo, (Cold Spring Harbor Laboratory cells and, thereby, reduce side effects. Press, Cold Spring Harbor, N.Y., 1986). 0520. The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage EXEMPLIFICATION for use in humans. The dosage of Such compounds may lie within a range of circulating concentrations that include the 0524. The invention now being generally described, it will EDs with little or no toxicity. The dosage may vary within be more readily understood by reference to the following this range depending upon the dosage form employed and the examples which are included merely for purposes of illustra route of administration utilized. For any compound, the thera peutically effective dose can be estimated initially from cell tion of certain aspects and embodiments of the present inven culture assays. A dose may beformulated in animal models to tion, and are not intended to limit the invention in any way. achieve a circulating plasma concentration range that includes the ICs (i.e., the concentration of the test compound Example 1 that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to Identification of Sirtuin Modulators Using SIRT1 more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance 0525. A fluorescence polarization or mass spectrometry liquid chromatography. based assay was used to identify modulators of SIRT1 activ ity. The same assay may be used to identify modulators of any 6. Kits sirtuin protein. The fluorescence polarization assays utilizes 0521. Also provided herein are kits, e.g., kits for therapeu one of two different peptides based on a fragment of p53, a tic purposes or kits for modulating the lifespan of cells or known sirtuin deacetylation target. Compounds were tested modulating apoptosis. A kit may comprise one or more sir using a Substrate containing peptide 1 having 20 amino acid tuin-modulating compounds, e.g., in premeasured doses. A residues as follows: Ac-EE-K(biotin)-GQSTSSHSK(Ac) kit may optionally comprise devices for contacting cells with NleSTEG-K(MR121)-EE-NH. (SEQ ID NO: 1) wherein the compounds and instructions for use. Devices include K(biotin) is a biotinolated lysine residue, K(Ac) is an acety Syringes, stents and other devices for introducing a sirtuin lated lysine residue, Nle is norleucine and K(MR121) is a modulating compound into a subject (e.g., the blood vessel of lysine residue modified by an MR121 fluorophore. This pep a subject) or applying it to the skin of a Subject. tide is labeled with the fluorophore MR121 (excitation 635 0522. Another type of kit contemplated by the invention nm/emission 680 nm) at the C-termini and biotin at the N-ter are kits for identifying sirtuin-modulating compounds. Such mini. The sequence of the peptide substrates are based on p53 kits contain (1) a sirtuin or sirtuin-containing material and (2) with several modifications. In particular, all arginine and leu a sirtuin-modulating compound of the invention, which are in cine residues other than the acetylated lysine residues have separate vessels. Such kits can be used, for example, to per replaced with serine so that the peptides are not susceptible to form a competition-type assay to test other compounds (typi trypsin cleavage in the absence of deacetylation. In addition, cally provided by the user) for sirtuin-modulating activity. In the methionine residues naturally present in the sequences certain embodiments, these kits further comprise means for determining sirtuin activity (e.g., a peptide with an appropri have been replaced with the norleucine because the methion ate indicator, Such as those disclosed in the Exemplification). ine may be susceptible to oxidation during synthesis and 0523 The practice of the present methods will employ, purification. As an alternative Substrate in the assay, the fol unless otherwise indicated, conventional techniques of cell lowing peptide 2 has also been used: Ac-EE-K(biotin)- biology, cell culture, molecular biology, transgenic biology, GQSTSSHSK(Ac)NleSTEG-K(5TMR)-EE-NH2 (SEQ ID microbiology, recombinant DNA, and immunology, which NO: 2) wherein K(Ac) is an acetylated lysine residue and Nle are within the skill of the art. Such techniques are explained is a norleucine. The peptide is labeled with the fluorophore fully in the literature. See, for example, Molecular Cloning A 5TMR (excitation 540 nm/emission 580 nm) at the C-termi Laboratory Manual, 2" Ed., ed. by Sambrook, Fritsch and nus. The sequence of the peptide Substrate is also based on Maniatis (Cold Spring Harbor Laboratory Press: 1989); DNA p53 with several modifications. In addition, the methionine Cloning, Volumes I and II (D. N. Glover ed., 1985); Oligo residue naturally present in the sequence was replaced with nucleotide Synthesis (M. J. Gait ed., 1984); Mullis et al. U.S. the norleucine because the methionine may be susceptible to Pat. No. 4,683, 195; Nucleic Acid Hybridization (B. D. oxidation during synthesis and purification. US 2009/0143376 A1 Jun. 4, 2009 54

0526. The peptide substrates were exposed to a sirtuin (SEQ ID NO: 2) wherein K(Ac) is an acetylated lysine resi protein in the presence of NAD" to allow deacetylation of the due and Nle is a norleucine. The peptide is labeled with the Substrate and render it sensitive to cleavage by trypsin. fluorophore 5TMR (excitation 540 nm/emission 580 inn) at Trypsin was then added and the reaction was carried to the C-terminus. The sequence of the peptide substrate is completion (i.e., the deacetylated Substrate is cleaved) releas based on p53 with several modifications. In addition, the ing the MR121 or 5TMR fragment. Streptavidin is then added methionine residue naturally present in the sequence was to the reaction where it can bind both the uncleaved substrate replaced with the norleucine because the methionine may be (i.e., any remaining acetylated Substrate) and the non-fluo Susceptible to oxidation during synthesis and purification. rescent portion of the cleaved peptide substrate (i.e., the biotin 0532. The mass spectrometry assay is conducted as fol containing fragment). The fluorescence polarization signal lows: 0.5uM peptide substrate and 120 uM BNAD" is incu observed for the full length peptide substrates bound to bated with 10 nM SIRT1 for 25 minutes at 25°C. in a reaction streptavidin was higher than the fluorescence polarization buffer (50 mM Tris-acetate pH 8, 137 mM NaCl, 2.7 mM signal observed for the released MR121 or 5TMR C-terminal KC1, 1 mM MgCl, 5 mM DTT, 0.05% BSA). Test com fragment. In this way, the fluorescence polarization obtained pounds may be added to the reaction as described above. The is inversely proportional to the level of deacetylation (e.g., the SirT1 gene is cloned into a T7-promoter containing vector signal is inversely proportional to the activity of the sirtuin and transformed into BL21 (DE3). After the 25 minute incu protein). Results were read on a microplate fluorescence bation with SIRT1, 10uI of 10% formic acid is added to stop polarization reader (Molecular Devices Spectramax MD) the reaction. Reactions are sealed and frozen for later mass with appropriate excitation and emission filters. spec analysis. Determination of the mass of the Substrate 0527 The fluorescence polarization assays using peptide peptide allows for precise determination of the degree of 1 is conducted as follows: 0.5 M peptide substrate and 150 acetylation (i.e. starting material) as compared to deacety uM BNAD" is incubated with 0.1 ug/mL of SIRT1 for 60 lated peptide (product). minutes at 37° C. in a reaction buffer (25 mM Tris-acetate 0533. For the mass spectrometry based assay, a control for pH8, 137 mM Na—Ac, 2.7 mM K—Ac, 1 mM Mg Ac, inhibition of sirtuin activity is conducted by adding 1 uL of 0.05% Tween-20, 0.1% Pluronic F127, 10 mM CaCl, 5 mM 500 mM nicotinamide as a negative control at the start of the DTT, 0.025% BSA, 0.15 mM Nicotinamide). Test com reaction (e.g., permits determination of maximum sirtuin pounds were solubilized in DMSO and added to the reaction inhibition). A control for activation of sirtuin activity is con ducted using 10 nM of sirtuin protein, with 1 uL of DMSO in at 11 concentrations ranging from 0.7 LM to 100 uM. place of compound, to determine the amount of deacetylation 0528 Fluorescence polarization assays using peptide 2 is of the Substrate at a given timepoint within the linear range of conducted as follows: 0.5uM peptide substrate and 120 uM the assay. This timepoint is the same as that used for test |BNAD" were incubated with 3 nM SIRT1 for 20 minutes at compounds and, within the linear range, the endpoint repre 25°C. in a reaction buffer (25 mM Tris-acetate pH8, 137 mM sents a change in Velocity. Na—Ac, 2.7 mMK Ac, 1 mM Mg Ac, 0.05% Tween-20, 0534 For each of the above assays, SIRT1 protein was 0.1% Pluronic F127, 10 mM CaCl, 5 mM DTT, 0.025% expressed and purified as follows. The SirT1 gene was cloned BSA). Test compounds 19-56 were solubilized in DMSO and into a T7-promoter containing vector and transformed into added to the reaction at 10 concentrations ranging from 300 BL21 (DE3). The protein was expressed by induction with 1 uM to 0.15uM in three-fold dilutions. mM IPTG as an N-terminal His-tag fusion protein at 18°C. 0529. After the incubation with SIRT1, nicotinamide was overnight and harvested at 30,000xg. Cells were lysed with added to the reaction to a final concentration of 3 mM to stop lysozyme in lysis buffer (50 mM Tris-HCl, 2 mM Tris(2- the deacetylation reaction and 0.5 ug/mL of trypsin was carboxyethyl phosphine (TCEP), 10 uM ZnCl2, 200 mM added to cleave the deacetylated substrate. The reaction was NaCl) and further treated with sonication for 10 min for incubated for 30 minutes at 37° C. in the presence of 1 LM complete lysis. The protein was purified over a Ni-NTA col streptavidin. Fluorescent polarization was determined at umn (Amersham) and fractions containing pure protein were excitation (650 nm) and emission (680 nm) wavelengths. The pooled, concentrated and run overa sizing column (Sephadex level of activity of the sirtuin protein in the presence of the S20026/60 global). The peak containing soluble protein was various concentrations of test compound is then determined collected and run on an Ion-exchange column(MonoO). Gra and may be compared to the level of activity of the sirtuin dient elution (200 mM-500 mM. NaCl) yielded pure protein. protein in the absence of the test compound, and/or the level This protein was concentrated and dialyzed against dialysis of activity of the sirtuin proteins in the negative control (e.g., buffer (20 mM Tris-HCl, 2 mMTCEP) overnight. The protein level of inhibition) and positive control (e.g., level of activa was aliquoted and frozen at -80°C. until further use. tion) described below. 0535 Sirtuin modulating compounds that activated SIRT1 0530 For the Fluorescence Polarization assays, a control were identified using the assay described above and are for inhibition of sirtuin activity is conducted by adding 1 uL shown below in Table 3. Sirtuin modulating compounds that of 500 mM nicotinamide as a negative control at the start of inhibited SIRT1 were identified using the assay described the reaction (e.g., permits determination of maximum sirtuin above and are shown below in Table 4. The EDso values for inhibition). A control for activation of sirtuin activity was the activating compounds are represented by A (EDso-z50 conducted using 3 nM of sirtuin protein, with 1 uL of DMSO uM), B (EDso-51-100 uM), C (EDso-101-150 uM), and D in place of compound, to reach baseline deacetylation of the (EDso-c 150 uM). The EDs of resveratrol for activation of Substrate (e.g., to determine normalized sirtuin activity). SIRT1 is 16 M. Similarly, the ICs values for the inhibiting 0531. The mass spectrometry based assay utilizes a pep compounds are represented by A (ICso C50 uM), B tide having 20 amino acid residues as follows: Ac-EE-K (ICs51-100 uM), G (ICs-101-150 uM), and D (Cs >150 (biotin)-GQSTSSHS-K(Ac)-N1e-STEG-K(5TMR)-EE-NH, uM). US 2009/0143376 A1 Jun. 4, 2009 55

TABLE 3

COMPOUND NO STRUCTURE EDso

1

N NH

N H

NH N NH2

HN

3

H N e SN N f O HN N

N 4 S -C H O - S

NH2

5 is S NH O US 2009/0143376 A1 Jun. 4, 2009 56

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

6 c A. S

HN S

O

OH

O

7 C

8 D

9 A.

10 US 2009/0143376 A1 Jun. 4, 2009 57

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 11 1. B O) or C 12 C2,S 4. or B 13 rO C NŠs Nu

2. O O 15 NH2 B

N

16 B US 2009/0143376 A1 Jun. 4, 2009

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

17 O

NH

2 N

e NH

18 NH2

HN

FN

Ne NH

HN

19

N US 2009/0143376 A1 Jun. 4, 2009 59

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

21 F

O NH Bra l O

NH OH 22 \

O

23 ClN1 N

24 1.

25 CON N H US 2009/0143376 A1 Jun. 4, 2009

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 26 NH2 C

HN O

21 2N. s-N/ N H

27 NH2 B

O O HN NH N 7 2 f N S NH

28 C

HO O US 2009/0143376 A1 Jun. 4, 2009 61

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

29 D

HO O ? O N

/ ,N , N2O S- I

30 B

HO O HN O N

/ (N , S. >

31 D

HO O HN O N / 2 N S. US 2009/0143376 A1 Jun. 4, 2009

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

32 B

HO O O HN 2N N 2 ( f N S. NH

33 H O C N

S. NH2

34 H O C N US 2009/0143376 A1 Jun. 4, 2009 63

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

35 HN

36

2N. )- N / l N

37

O)-() - US 2009/0143376 A1 Jun. 4, 2009 64

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

38 D

39 O C HN 21Ne.N. / N-N/ O

NH O O OH CS /

40 NH C

NH O 21Ne.N. / N-N/ O US 2009/0143376 A1 Jun. 4, 2009 65

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

41 C

goN1SN H

O

NH NS

42 B O

H N 21 N N C N S. N

HN O

NH US 2009/0143376 A1 Jun. 4, 2009 66

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

43 A. O

H N 21 NN N C N1SN

HN O

s'sNH

2-

44 C O

H N 21 NN N / N1SN

HN O

s'sNH US 2009/0143376 A1 Jun. 4, 2009 67

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

45 B O

H N

21 NN N N/ N s \

HN O

SSNH

-2

46 Q B NH a NN N N1's N SSHN O ON US 2009/0143376 A1 Jun. 4, 2009 68

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

47 B

H N 21 NN ) O N N N1SN

HN O

NH

2-

48 C

H N 21 N N NN N s NS C s'sHN O US 2009/0143376 A1 Jun. 4, 2009 69

3-continued

COMPOUND NO STRUCTURE EDso

49

N1's21 NSCO H

50

51 US 2009/0143376 A1 Jun. 4, 2009 70

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

52 C

N1's N H

N" O

53 C

21 )- O) US 2009/0143376 A1 Jun. 4, 2009 71

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

S4 D

gooN1SN H

N

NSN H

55 D

R. 7No

N1SNCity ( )?H Ol r HN O N

N O 21 N1 N N N1SN J US 2009/0143376 A1 Jun. 4, 2009 72

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

57 D

O 21 NN N N N1SN

O s O H 58 O A. NH O 21 N N N)-N O N1's N

HN O

O 59 O C NH O O 21 NN N )-O\ N1SN

HN O US 2009/0143376 A1 Jun. 4, 2009 73

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

60

NH O 21 N N < N N S N

O

61 D

H N

O

62 O A. S No C O

63 N NA US 2009/0143376 A1 Jun. 4, 2009

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

64 N B

65

( ) \N H N O N H

HO O 8 or 91 / Y. O \

N1's N 92 / Y. US 2009/0143376 A1 Jun. 4, 2009 75

TABLE 3-continued

COMPOUND NO STRUCTURE EDso

93 NN

94 CF CF

95

96 US 2009/0143376 A1 Jun. 4, 2009 76

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 97 | Y O

H N F 21 NN N CF N1's N 98 N

H N F 21 N N N1SN

99 CF CF

100 US 2009/0143376 A1 Jun. 4, 2009 77

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 101 | Y O O N

H N 21 NY /N N1's N S 102 | Y

103 / N US 2009/0143376 A1 Jun. 4, 2009 78

3-continued

COMPOUND NO STRUCTURE EDso 104 | Y

s ( )

105 | Yo US 2009/0143376 A1 Jun. 4, 2009 79

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 106 | Yo

O NHS. O

107 / N O \

O

N1's N

108 / Y. US 2009/0143376 A1 Jun. 4, 2009 80

TABLE 3-continued

COMPOUND NO STRUCTURE EDso 109 N

H N O 21 N N N1SN 110 CF CF / N\ /O O 21 NN N O N s N

TABLE 4 -continued

COMPOUND NO STRUCTURE ICso

84 O A. H

O O 21 NN N N1SN O NH NHHN Example 2 O 21 NN N Identification of Sirtuin Modulators Using SIRT3 N1's N 0536 A fluorescence polarization assay was used to iden tify modulators of SIRT3 activity. The same assay may be used to identify modulators of any sirtuin protein. The assay Synthesis of Compounds 91 through 110 is generally utilizes a peptide Substrate based on a fragment of Histone described in Angew Chemie, Int. Ed. 37 (16), 2234-37, 1998 H4, a known sirtuin deacetylation target. The Substrate con and as shown below: tains a peptide having 14 amino acid residues as follows: Biotin-GASSHSK(Ac)VLK(MR121) (SEQ ID NO: 3) wherein K(Ac) is an acetylated lysine residue. The peptide is labeled with the fluorophore MR121 (excitation 635 nm/emission 680 nm) at the C-terminus and biotin at the N-terminus. NN He CHO 0537. The peptide substrate is exposed to a sirtuin protein in the presence of NAD" to allow deacetylation of the sub 21 NH2 HClO4, MeOH strate and render it sensitive to cleavage by trypsin. Trypsin is then added and the reaction is carried to completion (i.e., the deacetylated substrate is cleaved) releasing the MR121 frag US 2009/0143376 A1 Jun. 4, 2009

ment. Streptavidin is then added to the reaction where it can cleave the deacetylated substrate. The reaction is incubated bind both the uncleaved substrate (i.e., any remaining acety for 30 minutes at 37°C. in the presence of 1 mM streptavidin. lated substrate) and the non-fluorescent portion of the cleaved Fluorescent polarization is determined at excitation (650 nm) peptide Substrate (i.e., the biotin containing fragment). The and emissions (680 nm) wavelengths. The level of activity of the sirtuin protein in the presence of the various concentra fluorescence polarization signal observed for the full length tions of test compound are then determined and may be com peptide substrate bound to streptavidin is higher than the pared to the level of activity of the sirtuin protein in the fluorescence polarization signal observed for the released absence of the test compound, and/or the level of activity of MR121 C-terminal fragment. Therefore, the fluorescence the sirtuin proteins in the negative control (e.g., level of inhi polarization obtained is inversely proportional to the level of bition) and positive control (e.g., level of activation) deacetylation (e.g., the signal is inversely proportional to the described below. activity of the sirtuin protein). Results are read on a micro 0539 A control for inhibition of sirtuin activity is con plate fluorescence polarization reader (Molecular Devices ducted by adding 30 mM nicotinamide at the start of the Spectramax MD) with appropriate excitation and emission reaction (e.g., permits determination of maximum sirtuin filters. inhibition). A control for activation of sirtuin activity is con 0538. The fluorescence polarization assays may be con ducted using 0.5 g/mL of sirtuin protein to reach baseline ducted as follows: 0.5 M peptide substrate and 50 LM deacetylation of the Substrate (e.g., to determine normalized |BNAD" is incubated with 2 nM of SIRT3 for 60 minutes at sirtuin activity). 37°C. in a reaction buffer (25 mM Tris-acetate pH8, 137 mM 0540 Sirtuin modulating compounds that activated SIRT3 Na—Ac, 2.7 mM K—Ac, 1 mM Mg Ac, 0.1% Pluronic were identified using the assay described above and are F127, 10 mM CaCl, 1 mM TCEP 0.025% BSA). Test com shown below in Table 5. Sirtuin modulating compounds that pounds are solubilized in DMSO and are added to the reaction inhibited SIRT3 were identified using the assay described at 11 concentrations ranging from 0.7 LM to 100 uM. The above and are shown below in Table 6. The EDso values for SIRT3 protein used in the assays corresponded to amino acid the activating compounds are represented by A (EDso-z50 residues 102-399 of human SIRT3 with an N-terminal His uM), B (EDso-51-100 uM), C (EDso-101-150 uM), and D tag. The protein was overexpressed in E. coli and purified on (EDso-c 150 uM). The EDs of resveratrol for activation of a nickel chelate column using standard techniques. After the SIRT1 is 16 M. Similarly, the ICs values for the inhibiting 60 minute incubation with SIRT3, nicotinamide is added to compounds are represented by A (ICso C50 uM), B the reaction to a final concentration of 3 mM to stop the (IC=51-100 uM), C (IC=101-150 uM), and D deacetylation reaction and 0.5 g/mL of trypsin is added to (ICs >150 uM).

TABLE 5

COMPOUND NO STRUCTURE EDso

C is S NH O

O --- OH

NA

HN S

O

OH US 2009/0143376 A1 Jun. 4, 2009 82

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

14 N

N H es

29

HO

O HN O

21Ne.N. / N-N? O NH NEO O

30 NA

HO

O HN O

21Ne.N. / N-N/ \ 31 NA

HO

O HN O 21Ne.N. / N-N/ NH NEO US 2009/0143376 A1 Jun. 4, 2009

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

32 A.

HO

O HN O

21 2N N-N/ EN NH

37 O OH C

HN O

43 US 2009/0143376 A1 Jun. 4, 2009 84

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

45 D O

H N 21 NN N / N S s \

HN O

SSNH

-2

67 D

HO

O HN O

21Ne.N. / N-N/ O NH NEO O/

68 NA

HO

O HN O O 21Ne.N. N-N/ US 2009/0143376 A1 Jun. 4, 2009 85

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

69 B

HO

O HN O O 21 2N. N-N/ NH

70 D

HO

O HN O

21 2N N-N/ EN NH

71 O HO O B N Nt I O HN O US 2009/0143376 A1 Jun. 4, 2009

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

72 B

HO

O HN O O

21 2N N-N/ SN NH

73 B

HO

O HN O O

21 2N. / N-N/ O NH NEO / O

74 D

HO

O HN O O

21 2N. s-N/ EN US 2009/0143376 A1 Jun. 4, 2009 87

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

75 HO

76 HO

---HN O

21Ne.N. / N-N/ O NH NtEO O/

77 O OH NA

HN O

21Ne.N. US 2009/0143376 A1 Jun. 4, 2009

TABLE 5-continued

COMPOUND NO STRUCTURE EDso

78 O OH C

HN O

79 O OH B

HN O

21 2N. N-N/ F HN

HO O N N NH y

81 O B

HN 21Ne.N. N-N/ EN NH O OH ( ) US 2009/0143376 A1 Jun. 4, 2009 89

TABLE 5-continued

COMPOUND NO STRUCTURE EDso 82 --CC () NA 83 NH2 D Oy (S- C HN N

TABLE 6

COMPOUND NO STRUCTURE ICso

85 NA OU---

O

86 O B HN-( ) { N S 7 HN -CIOC X- NH) ()- O

87 C US 2009/0143376 A1 Jun. 4, 2009 90

TABLE 6-continued

COMPOUND NO STRUCTURE ICso 88 A.

r NH Q NH O 21 NN / N N N s \

89 Ca C N O l YH O 21 NN N1SNN NHX (y1 O N-O

90 NA

O NH

NH NH O 4N-V N1's N

Equivalents by reference in their entirety as if each individual publication 0541. The present invention provides among other things or patent was specifically and individually indicated to be sirtuin-activating compounds and methods of use thereof. incorporated by reference. In case of conflict, the present While specific embodiments of the subject invention have application, including any definitions herein, will control. been discussed, the above specification is illustrative and not 0543. Also incorporated by reference in their entirety are restrictive. Many variations of the invention will become any polynucleotide and polypeptide sequences which refer apparent to those skilled in the art upon review of this speci ence an accession number correlating to an entry in a public fication. The full scope of the invention should be determined database, such as those maintained by The Institute for by reference to the claims, along with their full scope of Genomic Research (TIGR) (www.tigr.org) and/or the National Center for Biotechnology Information (NCBI) equivalents, and the specification, along with Such variations. (www.ncbi.nlm.nih.gov). INCORPORATION BY REFERENCE 0544. Also incorporated by reference are the following: 0542 All publications and patents mentioned herein, PCT Publications WO 2005/002672; 2005/002555; and including those items listed below, are hereby incorporated 2004/O16726. US 2009/0143376 A1 Jun. 4, 2009 91

1. A compound represented by Structural Formula (I): 2. A compound represented by Structural Formula (I):

(I) (I)

or a salt thereof, wherein, as valence permits: Ring A is optionally substituted; or a salt thereof, wherein, as valence permits: L is absent, substituted or unsubstituted phenylene, substi Ring A is optionally substituted; tuted or unsubstituted —O-phenylene, substituted or L is absent, substituted or unsubstituted phenylene, substi unsubstituted thienylene, substituted or unsubstituted tuted or unsubstituted —O-phenylene, substituted or pyrazolylene, substituted or unsubstituted benzothiaz unsubstituted thienylene, substituted or unsubstituted olylene, NR , —C(O)C) , —C(O)NR , pyrazolylene, substituted or unsubstituted benzothiaz —NRC(O) , NR, C(O) NRs , S , olylene, -C(O)O , —C(O)NR , —NRC(O) , —CHR=CHR or -CHR C(O)–: NR, C(O) NRs —S , —CHR=CHR, or L' is absent, substituted or unsubstituted phenylene, sub —CHR C(O)–: stituted or unsubstituted —O-phenylene, substituted or L' is absent, substituted or unsubstituted phenylene, sub unsubstituted thienylene, substituted or unsubstituted stituted or unsubstituted—O-phenylene, substituted or pyrazolylene, substituted or unsubstituted benzothiaz unsubstituted thienylene, substituted or unsubstituted olylene, substituted or unsubstituted indenedionylene, pyrazolylene, substituted or unsubstituted benzothiaz —C(O)C , C(O)NR , —NRC(O)—, NR C(O) NRs , S- CHR=CHR or olylene, -C(O)O , —C(O)NR , —NRC(O) , —CHRs C(O) provided that at least one of Land L' NR, C(O) NRs , —S , —CHR=CHR, or is substituted or unsubstituted phenylene, substituted or - CHRs C(O) , provided that at least one of Land L' unsubstituted -O-phenylene, substituted or unsubsti is substituted or unsubstituted phenylene, substituted or tuted thienylene, substituted or unsubstituted pyra unsubstituted -O-phenylene, substituted or unsubsti Zolylene, substituted or unsubstituted benzothiaz tuted thienylene, substituted or unsubstituted pyra olylene, substituted or unsubstituted indenedionylene, Zolylene, substituted or unsubstituted benzothiaz —NR , —C(O)C) , —C(O)NR , NRC(O) , olylene. —C(O)C) , —C(O)NR, , – NRC(O) , NR, C(O) NRs ,—S , —CHR=CHR, or NR, C(O) NRs —S , —CHR=CHR, or —CHRs C(O)–: - CHRs C(O) : R is absent, -H, -NRRs —NC(O)Rs, ORs, naph R is absent, -H, -NRRs —NC(O)Rs, ORs, naph thyl or a heterocyclic group, provided that L and Rare thyl or a heterocyclic group, provided that L and R are not both absent unless X is N: not both absent unless X is N: R is -H, unsubstituted alkyl, - NRRs, NRC(O)Rs. R is -H, unsubstituted alkyl, - NRRs, NRC(O)Rs. —ORs, substituted or unsubstituted phenyl, naphthyl or —ORs, naphthyl or a heterocyclic group; a heterocyclic group: R is -H, -NRRs, NC(O)Rs –ORs or a substituted R is -H, -NRRs, NC(O)Rs —ORs or a substituted or unsubstituted heterocyclic group, or R and R, taken or unsubstituted heterocyclic group, or RandR, taken together with the atoms to which they are attached, form together with the atoms to which they are attached, form an optionally substituted heterocyclic group, or R is an optionally substituted heterocyclic group, or R is absent when Z is O or S: absent when Z is O or S: Ra and Rs are independently —H, a substituted or unsub Ra and Rs are independently —H, a substituted or unsub stituted alkyl group, a substituted or unsubstituted aryl stituted alkyl group, a substituted or unsubstituted aryl group or a substituted or unsubstituted non-aromatic group or a substituted or unsubstituted non-aromatic heterocyclic group; heterocyclic group; R. R., and Rs are independently selected from the group R. R-7 and Rs are independently selected from the group consisting of halogen. —OR, CN, CO.R. consisting of halogen, —OR. -CN. COR, —OCOR - OCOR —C(O)NRRs, OC(O) —OCOR —OCOR –C(O)NRRs, OC(O) NRRs. -C(O)R. —COR. —SR - OSOH, NRRs. -C(O)R COR. —SR –OSOH, —S(O),R, S(O), OR —S(O)NRRs, NRRs. —S(O), R - S(O), OR - S(O)NRRs, NRRs, - NRC(O)CRs, NRC(O)Rs and - NO; NRC(O)ORs, NRC(O)Rs and NO; W is C or N: W is C or N: X is C or N: X is C or N: Y is C or N: Y is C or N: Zis C, N, O or S. provided that at least two of W, X, Y and Z is C, N, O or S, provided that at least two of W, X, Y and Zare C; and Zare C; and n is 1 or 2. n is 1 or 2. US 2009/0143376 A1 Jun. 4, 2009 92

3. A compound represented by Structural Formula (Ia): wherein: Z is selected from O or S. Ro is selected from —H. —C(O)—N(Rao)(Rs). —S(O) (Ia) N(R)(Rs), or —CH2—N(Rao)(Rs); wherein each of Rao and Rso is independently selected from —H. —C-C straight or branched alkyl, —(C-C straight or branched alkyl)-N(CH), —(C-C straight or branched alkyl)-heterocyclyl. —(C-C straight or R11 branched alkyl)-alkylheterocyclyl, or wherein Rao and YNH Rso taken together with the Natom to which they are bound form a 5-6 membered heterocyclic ring that is 21 NN N optionally Substituted with —(C-C straight or branched alkyl), and wherein at least one of Rao or Rso is N1SN s not H. R is selected from —C-C straight or branched alkylene R10 or —C(O)—: each of R and R is independently selected from —H or —(C-C straight or branched alkyl), or RandR are wherein: taken together to form a benzene ring that is substituted Ro is selected from —H. —C(O)—N(Rao)(Rs). —S(O) with up to two substituents independently selected from N(R)(Rs), or —CH2—N(Ra)(Rs); wherein each —(C-C straight or branched alkyl). —CF orhalo; and of Rao and Rso is independently selected from —H. ring K is substituted with up to three substituents indepen —C-C straight or branched alkyl, —(C-C straight or dently selected from halo. —CF, —O—(C-C straight branched alkyl)-N(CH), —(C-C straight or or branched alkyl). —S-(C-C straight or branched branched alkyl)-heterocyclyl. —(C-C straight or alkyl), —N(Rao)(Rs). —S(O) N(Rao)(Rso), hetero branched alkyl)-alkylheterocyclyl, or wherein Rao and cyclyl, (C-C straight or branched alkyl)-heterocyclyl, Rso taken together with the Natom to which they are —O—(C-C straight or branched alkyl)-heterocyclyl, bound form a 5-6 membered heterocyclic ring that is —S—(C-C straight or branched alkyl)-heterocyclyl, optionally substituted with —(C-C straight or or is optionally fused to a 5-6 membered heterocyclyl or branched alkyl), and wherein at least one of Rao or Rso is heteroaryl, wherein any heterocyclyl or heteroaryl is not H; optionally substituted with—C-C straight or branched R is selected from —C-C straight or branched alkylene alkyl. or —C(O)—, and 10-13. (canceled) each of ring Kand ring E is independently substituted with 14. A compound represented by Structural Formula (II): up to three substituents independently selected from halo. —CF, —O—(C-C straight or branched alkyl). (II) —S—(C-C straight or branched alkyl). —N(R) (Rs), —S(O), N(Rao)(Rs), heterocyclyl, (C-C straight or branched alkyl)-heterocyclyl. —O—(C-C, straight or branched alkyl)-heterocyclyl. —S-(C-C straight or branched alkyl)-heterocyclyl, or is optionally fused to a 5-6 membered heterocyclyl or heteroaryl, wherein any heterocyclyl or heteroaryl is optionally sub stituted with —C-C straight or branched alkyl. or a salt thereof, wherein: 4-8. (canceled) Rings C, D and E are optionally substituted; and 9. A compound represented by Structural Formula (Ib): X is 0 or 1. 15. (canceled) 16. The compound of claim 14, wherein the compound is (Ib) represented by Structural Formula (III):

(III) O R11 w NH D s R12 Cc) () 21S s 3-kiN R13, or a salt thereof, wherein: Rings D and E are optionally substituted; and R10 R is —H, a Substituted or unsubstituted alkyl group, a substituted or unsubstituted aryl group or a substituted or unsubstituted non-aromatic heterocyclic group. US 2009/0143376 A1 Jun. 4, 2009

17. The compound of claim 16, wherein the compound is or a salt thereof, wherein: represented by Structural Formula (IV): Ring F is optionally substituted; L' is substituted or unsubstituted phenylene, substituted or unsubstituted thienylene. —C(O)O— —NRC(O) , (IV) —S , —CHR=CHR or —CHRs C(O)—: R is —H, unsubstituted alkyl, —NRRs —NRC(O)Rs. O R5N —ORs, substituted or unsubstituted phenyl or a hetero R4 --- NH cyclic group; R is —H. —NRRs —NC(O)Rs, —ORs or a heterocy clic group, or RandR, taken together with the atoms to Cy-K)C. which they are attached, form an optionally substituted heterocyclic group, or R is absent when Z is O or S; Ra and Rs are independently —H, a Substituted or unsub or a salt thereof, wherein: stituted alkyl group, a Substituted or unsubstituted aryl Ring E is optionally substituted; and group or a Substituted or unsubstituted non-aromatic Ra and Rs are independently —H, a substituted or unsub heterocyclic group; stituted alkyl group, a Substituted or unsubstituted aryl R. R., and Rs are independently selected from the group group or a Substituted or unsubstituted non-aromatic consisting of halogen, —OR, —CN, —COR, heterocyclic group. –OCOR - OCOR –C(O)NRRs, OC(O) 18-19. (canceled) NRRs —C(O)R. —COR —SR —OSOH, 20. The compound of claim 17, wherein the compound is —S(O),R, —S(O), OR —S(O)NRRs —NRRs, represented by Structural Formula (V): —NRC(O)ORs, NRC(O)Rs and - NO; Z is C, N, O or S; and n is 1 or 2. (V) 23. The compound of claim 22, wherein the compound is O represented by Structural Formula (VII): R4 NN1--> >< N-NH N (VII) nus/ \ / R3 A R 2: or a salt thereof, wherein: R. Rs and R are independently —H, a substituted or N unsubstituted alkyl group, a Substituted or unsubstituted aryl group or a substituted or unsubstituted non-aro or a salt thereof, wherein: matic heterocyclic group. Ring F is optionally substituted; 21. The compound of claim 20, wherein the compound is L' is substituted or unsubstituted phenylene, substituted or represented by Structural Formula (VI): unsubstituted thienylene. —C(O)O— —S , —CHR=CHR, or —CHRs C(O) ; (VI) R is —H, unsubstituted alkyl, —NRRs —NRC(O)Rs. Rs —ORs or a heterocyclic group; V R is —H. —NRRs —NC(O)Rs, —ORs or a heterocy NH O clic group, or RandR, taken together with the atoms to which they are attached, form an optionally substituted 21 N N NHX-R. heterocyclic group, or R is absent when Z is O or S; Ra and Rs are independently —H, a Substituted or unsub S s stituted alkyl group, a Substituted or unsubstituted aryl group or a Substituted or unsubstituted non-aromatic R4 heterocyclic group; YN O H R. R., and Rs are independently selected from the group consisting of halogen, —OR, —CN, —COR, –OCOR - OCOR –C(O)NRRs, OC(O) or a salt thereof, wherein: NRRs —C(O)R. —COR —SR —OSOH, R. Rs and Ro are independently —H, a Substituted or —S(O),R, —S(O), OR —S(O)NRRs —NRRs, unsubstituted alkyl group, a Substituted or unsubstituted —NRC(O)ORs, —NRC(O)Rs and —NO; aryl group or a substituted or unsubstituted non-aro Z is C, N, O or S; and matic heterocyclic group. n is 1 or 2. 22. A compound represented by Structural Formula (VII): 24. The compound of claim 22, wherein the compound is represented by Structural Formula (VIII): (VII) R3 M (VIII) Z R2, sy A L US 2009/0143376 A1 Jun. 4, 2009 94 or a salt thereof, wherein: or a salt thereof, wherein: Ring G is optionally substituted; Ring H is optionally substituted; L' is substituted or unsubstituted phenylene, substituted or L' is substituted or unsubstituted phenylene. —S or unsubstituted —O-phenylene, substituted or unsubsti —CHR=CHR, ; tuted thienylene, substituted or unsubstituted indenedio R is —NRRs —NRC(O)Rs or a heterocyclic group; nylene. —NRC(O)— —C(O)O— —S , R is —H, or R and R, taken together with the atoms to —CHR=CHR or —CHRs C(O)—: which they are attached, form an optionally substituted R is —H, unsubstituted alkyl, —NRRs —NRC(O)Rs. heterocyclic group; —ORs, substituted or unsubstituted phenyl or a hetero Ra and Rs are independently —H, a Substituted or unsub cyclic group; stituted alkyl group, a Substituted or unsubstituted aryl R and Rs are independently —H, a Substituted or unsub group or a Substituted or unsubstituted non-aromatic stituted alkyl group, a Substituted or unsubstituted aryl heterocyclic group; group or a Substituted or unsubstituted non-aromatic Re and R, are independently selected from the group con heterocyclic group; sisting of halogen, —OR —CN, —COR —OCOR, R. R., and Rs are independently selected from the group –OCOR –C(O)NRRs – OC(O)NRRs –C(O) consisting of halogen, —OR, —CN, —COR, R —COR —SR —OSOH, -S(O), R —S(O) –OCOR - OCOR –C(O)NRRs, OC(O) OR —S(O)NRRs, NRRs, NRC(O)CRs. NRRs —C(O)R. —COR —SR —OSOH, —NRC(O)Rs and —NO; and —S(O), R —S(O), OR —S(O)NRRs —NRRs, n is 1 or 2. —NRC(O)ORs, NRC(O)Rs and - NO; and 35-41. (canceled) n is 1 or 2. 42. The compound of claim 22, wherein the compound is 25. The compound of claim 22, wherein the compound is represented by Structural Formula (X): represented by Structural Formula (VIII): (X) (VIII) L - R3 S R2, I X-1/ N R2, N 2 -- or a salt thereof, wherein: R Ring G is optionally substituted; L' is substituted or unsubstituted phenylene, substituted or or a salt thereof, wherein: unsubstituted —O-phenylene, substituted or unsubsti Ring J is optionally Substituted; tuted thienylene, —C(O)O , —S , L is substituted or unsubstituted phenylene, substituted or —CHR=CHR, or —CHRs C(O) ; unsubstituted —O-phenylene, substituted or unsubsti R is —H, unsubstituted alkyl, - NRRs, NRC(O)Rs. tuted thienylene, substituted or unsubstituted pyra —ORs or a heterocyclic group; zolylene, substituted or unsubstituted benzothiaz R and Rs are independently —H, a Substituted or unsub olylene. —C(O)O , —C(O)NR —NRC(O)—, stituted alkyl group, a Substituted or unsubstituted aryl NR C(O) NRs —S , —CHR=CHR, or group or a Substituted or unsubstituted non-aromatic —CHR C(O)—: heterocyclic group; L' is substituted or unsubstituted phenylene, substituted or R. R., and Rs are independently selected from the group unsubstituted —O-phenylene, substituted or unsubsti consisting of halogen, —OR, —CN, —COR, tuted thienylene, substituted or unsubstituted pyra –OCOR - OCOR –C(O)NRRs –OC(O) zolylene, substituted or unsubstituted benzothiaz NRRs —C(O)R. —COR —SR —OSOH, olylene. —C(O)O , —C(O)NR —NRC(O)—, —S(O), R —S(O), OR —S(O)NRRs —NRRs, NR, C(O) NRs , —S , —CHR=CHR, or —NRC(O)ORs, —NRC(O)Rs and —NO; and —CHRs C(O)—: n is 1 or 2. R is —H. —NRRs —NC(O)Rs —ORs, naphthyl or a 26-33. (canceled) heterocyclic group; 34. The compound of claim 22, wherein the compound is R is —H, unsubstituted alkyl, —NRRs —NRC(O)Rs. represented by Structural Formula (IX): —ORs, naphthyl or a heterocyclic group; R is —H, NRRs, NC(O)Rs—ORs or a substituted or unsubstituted heterocyclic group; (IX) Ra and Rs are independently —H, a Substituted or unsub R3 stituted alkyl group, a Substituted or unsubstituted aryl \ R 22 group or a Substituted or unsubstituted non-aromatic heterocyclic group; 2 / R. R., and Rs are independently selected from the group consisting of halogen, —OR, —CN, —COR, –OCOR - OCOR –C(O)NRRs, OC(O) NRRs —C(O)R. —COR —SR —OSOH, US 2009/0143376 A1 Jun. 4, 2009 95

—S(O), R —S(O), OR —S(O)NRRs —NRRs, compound of any of claims 1-3, 9, 14, and 22, or a pharma —NRC(O)ORs, —NRC(O)Rs and —NO; and ceutically acceptable salt or prodrug thereof. n is 1 or 2. 70-74. (canceled) 43-48. (canceled) 75. A method for treating or preventing chemotherapeutic 49. A composition comprising a compound of any of induced neuropathy comprising administering to a Subject in claims 1-3, 9, 14, and 22, wherein the composition is pyro need thereofatherapeutically effective amount of at least one gen-free. compound of any of claims 1-3, 9, 14, and 22, or a pharma 50. A pharmaceutical composition comprising a pharma ceutically acceptable salt or prodrug thereof. ceutically acceptable carrier or diluent and a compound of 76. (canceled) any of claims 1-3, 9, 14, and 22. 77. A method for treating or preventing neuropathy asso 51. A packaged pharmaceutical comprising a compound of ciated with an ischemic event or disease comprising admin any of claims 1-3, 9, 14, and 22 and instructions for using the istering to a subject in need thereofatherapeutically effective compound to modulate a sirtuin. amount of at least one compound of any of claims 1-3, 9, 14, 52. A method for promoting survival of a eukaryotic cell and 22, or a pharmaceutically acceptable salt or prodrug comprising contacting the cell with at least one compound of thereof. any of claims 1-3, 9, 14, and 22, or a pharmaceutically accept 78. (canceled) able salt or prodrug thereof. 79. A method for treating or preventing a polyglutamine 53-57. (canceled) disease comprising administering to a Subject in need thereof 58. A method for treating or preventing a disease or disor a therapeutically effective amount of at least one compound der associated with cell death or aging in a Subject, compris of any of claims 1-3, 9, 14, and 22, or a pharmaceutically ing administering to a Subject in need thereof a therapeuti acceptable salt or prodrug thereof. cally effective amount of at least one compound of any of 80-81. (canceled) claims 1-3, 9, 14, and 22, or a pharmaceutically acceptable 82. A method for treating a disease or disorder in a subject salt or prodrug thereof. that would benefit from increased mitochondrial activity, 59. (canceled) comprising administering to a Subject in need thereofathera 60. A method for treating or preventing insulin resistance, peutically effective amount of at least one compound of any a metabolic syndrome, diabetes, or complications thereof, or of claims 1-3, 9, 14, and 22, or a pharmaceutically acceptable for increasing insulin sensitivity in a subject, comprising salt or prodrug thereof. administering to a subject in need thereof a therapeutically 83-88. (canceled) effective amount of at least one compound of any of claims 89. A method for enhancing motor performance or muscle 1-3, 9, 14, and 22, or a pharmaceutically acceptable salt or endurance, decreasing fatigue, or increasing recovery from prodrug thereof. fatigue, comprising administering to a subject in need thereof 61. A method for reducing the weight of a subject, or a therapeutically effective amount of at least one compound preventing weight gain in a Subject, comprising administer of any of claims 1-3, 9, 14, and 22, or a pharmaceutically ing to a subject in need thereof a therapeutically effective acceptable salt or prodrug thereof. amount of at least one compound of any of claims 1-3, 9, 14, 90-91. (canceled) and 22, or a pharmaceutically acceptable salt or prodrug 92. A method for treating or preventing a condition wherein thereof. motor performance or muscle endurance is reduced, compris 62. (canceled) ing administering to a subject in need thereof a therapeuti 63. A method for preventing the differentiation of a pre cally effective amount of at least one compound of any of adipocyte, comprising contacting the pre-adipocyte with at claims 1-3, 9, 14, and 22, or a pharmaceutically acceptable least one compound of any of claims 1-3, 9, 14, and 22, or a salt or prodrug thereof. pharmaceutically acceptable salt or prodrug thereof. 93. (canceled) 64. A method for prolonging the lifespan of a subject 94. A method for treating or preventing muscle tissue dam comprising administering to a Subject atherapeutically effec age associated with hypoxia or ischemia, comprising admin tive amount of at least one compound of any of claims 1-3, 9. istering to a subject in need thereofatherapeutically effective 14, and 22, or a pharmaceutically acceptable salt or prodrug amount of at least one compound of any of claims 1-3, 9, 14, thereof. and 22, or a pharmaceutically acceptable salt or prodrug 65. A method for treating or preventing a neurodegenera thereof. tive disorder in a Subject, comprising administering to a Sub 95. A method for increasing muscle ATP levels in a subject, ject in need thereof a therapeutically effective amount of at comprising administering to a Subject in need thereofathera least one compound of any of claims 1-3, 9, 14, and 22, or a peutically effective amount of at least one compound of any pharmaceutically acceptable salt or prodrug thereof. of claims 1-3, 9, 14, and 22, or a pharmaceutically acceptable 66. (canceled) salt or prodrug thereof. 67. A method for treating or preventing a blood coagulation 96-101. (canceled) disorderina Subject, comprising administering to a Subject in 102. A method for treating or preventing cancer in a Sub need thereofatherapeutically effective amount of at least one ject, comprising administering to a subject in need thereof a compound of any of claims 1-3, 9, 14, and 22, or a pharma therapeutically effective amount of at least one compound of ceutically acceptable salt or prodrug thereof. any of claims 1-3, 9, 14, and 22, or a pharmaceutically accept 68. (canceled) able salt or prodrug thereof. 69. A method for treating or preventing an ocular disease or 103. (canceled) disorder, comprising administering to a Subject in need 104. A method for stimulating weight gain in a Subject, thereof a therapeutically effective amount of at least one comprising administering to a Subject in need thereofathera US 2009/0143376 A1 Jun. 4, 2009 96 peutically effective amount of at least one compound of any at least one compound of any of claims 1-3, 9, 14, and 22, or of claims 1-3, 9, 14, and 22, or a pharmaceutically acceptable a pharmaceutically acceptable salt or prodrug thereof. salt or prodrug thereof. 106-111. (canceled) 105. A method for increasing the radiosensitivty or chemosensitivity of a cell comprising contacting the cell with ck