Senotherapeutic compounds

Abstract

The invention provides a compound of formula (I): or a salt thereof, wherein R1 has any of the values described in the specification, as well as compositions comprising a compound of formula (I). The compounds are useful as senolytic agents.

Description

[0001] SENOTHERAPEUTIC COMPOUNDS

[0002] CROSS-REFERENCE TO RELATED APPLICATION

[0003] This application claims priority to United States Provisional Application Number 63 / 527,369 that was filed on July 18, 2023. The entire content of the applications referenced above is hereby incorporated by reference herein.

[0004] STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH

[0005] This invention was made with government support under AG056278, AG043376, and AG062413 awarded by the National Institutes of Health. The government has certain rights in the invention

[0006] BACKGROUND OF THE INVENTION

[0007] Aging is often accompanied by declines in health and physiological functions, resulting in an increase in the prevalence of numerous chronic diseases. One of the critical hallmarks or underlying mechanisms of aging is cellular senescence (Lopez-Otin, C., et al., Cell 153, 1194- 1217 (2013); Kennedy, B.K., et al. Cell 159, 709-713 (2014); and Lopez-Otin, et al., Cell 186, 243-278 (2023)). Senescence is a phenomenon in which cells become incapable of proliferating under various cellular stress conditions, leading to a state of stable cell cycle arrest (Hayflick, L. & Moorhead, P.S. Exp Cell Res 25, 585-621 (1961)). With age, senescent cells accumulate in many tissues and produce a range of deleterious signals that can negatively affect neighboring cells. These signals, known as senescence-associated secretory phenotype (SASP), consist of diverse pro-inflammatory cytokines, chemokines, and proteases (Coppe, J.P., et al., Annu Rev Pathol 5, 99-118 (2010); and Di Micco, R., et al., Genes Dev 34, 1565-1576 (2020)). The SASP released by senescent cells can result in tissue dysfunction and deterioration of tissue and even turn more healthy cells senescent, thereby further driving aging and promoting many age-related diseases and pathologies (Birch, J. & Gil, J. Genes Dev 34, 1565-1576 (2020); Gorgoulis, V., et al. Cell 179, 813-827 (2019); and Wiley, C.D. & Campisi, J. Nat Metab 3, 1290-1301 (2021)). The use of transgenic mouse models, where senescent cells can be genetically ablated, has established a key role of senescent cells in driving aging and age-related diseases (Baker, D.J., et al. Nature 479, 232-236 (2011); and Baker, D.J., et al. Nature 530, 184-189 (2016)). Pharmacologically eliminating senescent cell burden using a class of drugs known as senolytics has been shown to be an effective strategy to extend healthspan and treat age-related diseases (Robbins, P.D., et al. Annu Rev Pharmacol Toxicol 61, 779-803 (2021); Prasnikar, E., Borisek, J. & Perdih, Ageing Res Rev 66, 101251 (2020); and Kirkland, J.L. & Tchkonia, T. J Intern Med 288, 518-536 (2020)).

[0008] Senolytics have emerged as an effective therapeutic approach to eliminate senescent cells and improve aging phenotypes and co-morbidities. Despite their promising potential, only a limited number of senolytics have been discovered. Fisetin has been demonstrated to reduce senescence, suppress age-related pathology, and extend healthspan in aged mice (Yousefzadeh, M.J., et al. EBioMedicine 36, 18-28 (2018)). However, its moderate potency and poor bioavailability have hindered its further clinical applications.

[0009] Currently there is a need for compounds that specifically induce apoptosis in senescent cells, termed senolytics. In particular, there is a need for senolytic compounds that have improved potency, lower mutagenic risk, and / or improved bioavailability. Such compounds would be useful for reducing the senescent cell burden that increases with aging and diseases, extending healthspan or lifespan, and / or treating age-related diseases.

[0010] SUMMARY OF THE INVENTION

[0011] In one aspect senolytic compounds that have improved potency, lower mutagenic risk, and / or improved bioavailability are provided.

[0012] Accordingly, a compound of formula I: or a salt thereof, wherein:

[0013] R1is isopropyl, tert-butyl, 3 -methylpropyl, 1 -methylpropyl, (C3-Ce)cycloalkyl, or phenyl, wherein any (C3-Ce)cycloalkyl and phenyl is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro is provided. Aa pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient is also provided.

[0014] A method for reducing the expression of one or more senescence markers in a cell comprising, contacting the cell with a compound of formula (I) or a salt thereof is also provided. The method can be carried out in vitro or in an animal.

[0015] A method for reducing senescence in an animal, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is also provided.

[0016] A method for extending the healthspan or lifespan of an animal, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is also provided.

[0017] A method for treating age-related disease in an animal, comprising administering a compound of formula (I) or a pharmaceutically acceptable salt thereof to the animal is also provided.

[0018] A method for increasing senotherapeutic activity in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal is also provided.

[0019] A compound of formula (I) or a pharmaceutically acceptable salt thereof for use in medical therapy is also provided.

[0020] A compound of formula I or a pharmaceutically acceptable salt thereof for reducing senescence, extending healthspan, treating an age-related disease, or increasing senotherapeutic activity is also provided.

[0021] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing senescence, extending healthspan, treating an age-related disease, or increasing senotherapeutic activity is also provided.

[0022] A method for treating a disease or condition in an animal selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bone marrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, tissue regeneration, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0023] A method for improving response to chemotherapy in an animal, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0024] A method for improving response to radiation therapy in an animal, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0025] A method for improving recovery of a cancer survivor, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the cancer survivor is also provided.

[0026] A method for improving cognition in an animal, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0027] A method for reducing the negative effects of tobacco use (past or present) in an animal, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0028] A method for reducing one or more symptoms associated with long-haul COVID-19 in an animal, comprising administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0029] A method comprising contacting an organ (e.g., an organ from an older doner) with a compound of formula (I) a pharmaceutically acceptable salt thereof to improve the characteristics of the organ for transplantation is also provided.

[0030] Aa method for improving bone healing in an animal comprising, administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided. A method for repairing damaged muscle in an animal comprising, administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0031] A method for improving tissue homeostasis comprising, contacting tissue with a compound of formula (I) a pharmaceutically acceptable salt thereof is also provided.

[0032] A method for improving immune response to a vaccine in an animal comprising, administering a compound of formula (I) a pharmaceutically acceptable salt thereof to the animal is also provided.

[0033] A compound of formula I or a pharmaceutically acceptable salt thereof for treating a disease or condition selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bone marrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, tissue regeneration, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection is also provided.

[0034] A compound of formula I or a pharmaceutically acceptable salt thereof for improving response to chemotherapy is also provided.

[0035] A compound of formula I or a pharmaceutically acceptable salt thereof for improving response to radiation therapy is also provided.

[0036] A compound of formula I or a pharmaceutically acceptable salt thereof for improving recovery of a cancer survivor is also provided.

[0037] A compound of formula I or a pharmaceutically acceptable salt thereof for improving cognition is also provided.

[0038] A compound of formula I or a pharmaceutically acceptable salt thereof for reducing the negative effects of tobacco use (past or present) is also provided. A compound of formula I or a pharmaceutically acceptable salt thereof for reducing one or more symptoms associated with long-haul COVID-19 is also provided.

[0039] A compound of formula I or a pharmaceutically acceptable salt thereof for improving the characteristics of an organ for transplantation is also provided.

[0040] A compound of formula I or a pharmaceutically acceptable salt thereof for improving bone healing is also provided.

[0041] A compound of formula I or a pharmaceutically acceptable salt thereof for repairing damaged muscle is also provided.

[0042] A compound of formula I or a pharmaceutically acceptable salt thereof for improving tissue homeostasis is also provided.

[0043] A compound of formula I or a pharmaceutically acceptable salt thereof for improving immune response to a vaccine is also provided.

[0044] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for treating a disease or condition selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bone marrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, tissue regeneration, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection in an animal is also provided.

[0045] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving response to chemotherapy in an animal is also provided.

[0046] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving response to radiation therapy in an animal is also provided.

[0047] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving recovery of a cancer survivor is also provided. The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving cognition in an animal is also provided.

[0048] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing the negative effects of tobacco use (past or present) in an animal is also provided.

[0049] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing one or more symptoms associated with long-haul COVID-19 in an animal is also provided.

[0050] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving the characteristics of an organ for transplantation is also provided.

[0051] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving bone healing in an animal is also provided.

[0052] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for repairing damaged muscle in an animal is also provided.

[0053] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving tissue homeostasis in an animal is also provided.

[0054] The use of a compound of formula I or a pharmaceutically acceptable salt thereof to prepare a medicament for improving immune response to a vaccine in an animal is also provided.

[0055] Processes and intermediates disclosed herein that are useful for preparing a compound of formula (I) or a salt thereof are also provided.

[0056] BRIEF DESCRIPTION OF DRAWINGS

[0057] Figs, la-le. (a) At 27-months of age, wild-type C57BL / 6J mice were administered vehicle, fisetin, SR29384 (Example 1), or SR31133 (Example 3) for 5 consecutive days by oral gavage. Mice were sacrificed two days after the last dose, (b-e) Tissues were collected and analyzed for changes in senescence SASP markers by RT-qPCR.

[0058] Figs. 2a-2c. (a) At 10-weeks of age, Erccl'^ progeria mice were administered SR29384 (Example 1) or vehicle for 5 consecutive weeks by oral gavage. Mice were sacrificed two days after the last dose, (b) Health assessments were performed weekly to score diverse age symptoms including tremor, kyphosis, dystonia, ataxia, gait disorder, hindlimb paralysis and forelimb grip strength, (c) Tissues were collected and analyzed for changes in senescence SASP markers by RT-qPCR

[0059] DETAILED DESCRIPTION OF THE INVENTION

[0060] The term “cycloalkyl” refers to a saturated or partially unsaturated (non-aromatic) all carbon ring having 3 to 8 carbon atoms (i.e., (Cs-Csjcycloalkyl). In one embodiment, cycloalkyl is (C3-C6)cycloalkyl. In one embodiment, cycloalkyl is (C4-C6)cycloalkyl. The cycloalkyl ring may comprise one or more (e.g., one or two) double bonds.

[0061] As used herein, the term "protecting group" refers to a substituent that is commonly employed to block or protect a particular functional group on a compound. For example, an "amino-protecting group" is a substituent attached to an amino group that blocks or protects the amino functionality in the compound. Suitable amino-protecting groups include acetyl, trifluoroacetyl, t-butoxycarbonyl (BOC), benzyloxycarbonyl (CBZ) and 9- fluorenylmethylenoxycarbonyl (Fmoc). Similarly, a "hydroxy-protecting group" refers to a substituent of a hydroxy group that blocks or protects the hydroxy functionality. Suitable protecting groups include acetyl and silyl. A "carboxy-protecting group" refers to a substituent of the carboxy group that blocks or protects the carboxy functionality. Common carboxy- protecting groups include phenyl sulfonylethyl, cyanoethyl, 2-(trimethylsilyl)ethyl, 2- (trimethylsilyl)ethoxymethyl, 2-(p-toluenesulfonyl)ethyl, 2-(p-nitrophenylsulfenyl)ethyl, 2- (diphenylphosphino)-ethyl, nitroethyl and the like. For a general description of protecting groups and their use, see P.G.M. Wuts and T.W. Greene, Greene's Protective Groups in Organic Synthesis 4thedition, Wiley-Interscience, New York, 2006.

[0062] As used herein a wavy line “ -~w ” that intersects a bond in a chemical structure indicates the point of attachment of the bond that the wavy bond intersects in the chemical structure to the remainder of a molecule.

[0063] The terms “treat”, “treatment”, or “treating” to the extent it relates to a disease or condition includes inhibiting the disease or condition, eliminating the disease or condition, and / or relieving one or more symptoms of the disease or condition. The terms “treat”, “treatment”, or “treating” also refer to both therapeutic treatment and / or prophylactic treatment or preventative measures, wherein the object is to prevent or slow down (lessen) an undesired physiological change or disorder, such as, for example, the development or spread of cancer. For example, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease or disorder, stabilized (i.e., not worsening) state of disease or disorder, delay or slowing of disease progression, amelioration or palliation of the disease state or disorder, and remission (whether partial or total), whether detectable or undetectable. “Treat”, “treatment”, or “treating,” can also mean prolonging survival as compared to expected survival if not receiving treatment. Those in need of treatment include those already with the disease or disorder as well as those prone to have the disease or disorder or those in which the disease or disorder is to be prevented. In one embodiment “treat”, “treatment”, or “treating” does not include preventing or prevention,

[0064] The phrase "therapeutically effective amount" or “effective amount” includes but is not limited to an amount of a compound of the that (i) treats or prevents the particular disease, condition, or disorder, (ii) attenuates, ameliorates, or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition, or disorder described herein.

[0065] The term “animal” as used herein includes mammals.

[0066] The term “mammal” as used herein refers to humans, higher non-human primates, rodents, domestic, cows, horses, pigs, sheep, dogs and cats. In one embodiment, the mammal is a human. The term “patient” as used herein refers to any animal including mammals. In one embodiment, the patient is a mammalian patient. In one embodiment, the patient is a human patient.

[0067] The compounds disclosed herein can also exist as tautomeric isomers in certain cases. Although only one delocalized resonance structure may be depicted, all such forms are contemplated within the scope of the invention.

[0068] It is understood by one skilled in the art that this invention also includes any compound claimed that may be enriched at any or all atoms above naturally occurring isotopic ratios with one or more isotopes such as, but not limited to, deuterium (2H or D). As a non-limiting example, a -CH3 group may be substituted with -CD3.

[0069] The pharmaceutical compositions of the invention can comprise one or more excipients. When used in combination with the pharmaceutical compositions of the invention the term “excipients” refers generally to an additional ingredient that is combined with the compound of formula (I) or the pharmaceutically acceptable salt thereof to provide a corresponding composition. For example, when used in combination with the pharmaceutical compositions of the invention the term “excipients” includes, but is not limited to: carriers, binders, disintegrating agents, lubricants, sweetening agents, flavoring agents, coatings, preservatives, and dyes.

[0070] Specific values listed below for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for the radicals and substituents. It is to be understood that two or more values may be combined. It is also to be understood that the values listed herein below (or subsets thereof) can be excluded.

[0071] A specific value for R1is isopropyl, tert-butyl, or 3 -methylpropyl.

[0072] A specific value for R1is (C3-Ce)cycloalkyl that is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro.

[0073] A specific value for R1is phenyl that is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro.

[0074] A specific value for R1is isopropyl, tert-butyl, 3 -methylpropyl, cyclopentyl, cyclohexyl, 1 -cyclohexenyl, phenyl, 2-hydroxyphenyl, 3 -hydroxyphenyl, 3 -methoxyphenyl, 4- m ethoxyphenyl, or 3 -trifluoromethylphenyl.

[0075] Processes for preparing compounds of formula I are provided as further embodiments of the invention.

[0076] In cases where compounds are sufficiently basic or acidic, a salt of a compound of formula I can be useful as an intermediate for isolating or purifying a compound of formula I. Additionally, administration of a compound of formula I as a pharmaceutically acceptable acid or base salt may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, a- ketoglutarate, and a-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts.

[0077] Salts may be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example calcium) salts of carboxylic acids can also be made. The compounds of formula I can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human patient in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes.

[0078] Thus, the present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained.

[0079] The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as com starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices. The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.

[0080] The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glyceryl esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelatin.

[0081] Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions.

[0082] Useful dosages of the compounds of formula I can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949. The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations; such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye. The invention will now be illustrated by the following non-limiting Examples.

[0083] EXAMPLES

[0084] Example 1. 2-(3,4-Dihydroxyphenyl)-6-isopropyl-4H-chromen-4-one (SR29384) a. 4-Isopropylphenyl acetate

[0085] To the mixture of 4-isopropylphenol (5.0g, 36.91 mmol) in acetic anhydride (5.2 mL, 55.37 mmol) was added 2 drops of cone. H2SO4 at room temperature. The completion of the reaction was monitored by analytical HPLC. The reaction mixture was concentrated, redissolved with EtOAc, washed with saturated NaHCCh, brine and dried over Na2SO4 to obtain the title compound with no further purification.JH NMR (400 MHz, CDCI3) 5 7.22 (d, J=8.4 Hz, 2H), 6.99 (d, J=8.4 Hz, 2H), 2.99-2.87 (m, 1H), 2.29 (s, 3H), 1.24 (d, J=6.9 Hz, 6H) b. l-(2-Hydroxy-5-isopropylphenyl)ethan-l-one

[0086] 4-Isopropylphenyl acetate (10.17 g, 57.06 mmol) was added slowly to

[0087] Trifluoromethanesulfonic acid (20 mL) at ice bath. The completion of reaction was monitored by analytical HPLC (2h). The reaction was poured into ice and extracted with EtOAc, washed with waier, saturated NaHCOs, and brine, and dried over Na2.SO4. The solvent was removed to obtain the crude which was purified via silica gel flash chromatography to obtain the title compound.XH NMR (400 MHz, CDCh) 5 12.29 (s, 1H), 7.64 (d, J=8.2 Hz, 1H), 6.83 (d, J=1.6 Hz, 1H), 6.77 (dd, J=8.2 Hz, 1.6 HZ, 1H), 2.99-2.87 (m, 1H), 2.60 (s, 3H), 1.24 (d, J=6.9 Hz, 6H) c. (E)-3-(3,4-bis(Benzyloxy)phenyl)-l-(2-hydroxy-5-isopropylphenyl)prop-2-en-l-one

[0088] To a mixture of l-(2-hydroxy-5-isopropylphenyl)ethan-l-one (0.28 g, 1.57 mmol) and 3,4-bis(benzyloxy)benzaldehyde (0.5g, 1.57 mmol) in EtOH (10 mL) was added NaOH solution (1.6 mL, 5 M in water, 8.0 mmol) and the reaction was stirred at room temperature for 2 days. The mixture was poured into ice water, extracted with EtOAc, washed with water, saturated NaHCOs, and brine, and dried over Na2SO4. The solvent was removed to obtain the crude which was purified via silica gel flash chromatography to obtain the title compound.JH NMR (400 MHz, CDCh) 5 7.47-7.26 (m, 14H), 7.14 (d, J=2.0 Hz, 1H), 6.09 (dd, J=8.2 Hz, 2.0 HZ, 1H), 6.92 (d, J=8.4 Hz, 1H), 6.54 (d, J=16.0 Hz, 1H), 5.21 (s, 2H), 5.19 (s, 2H), 2.24-2.16 (m, 1H), 0.96 (d, J=6.9 Hz, 6H) d. 2-(3,4-bis(Benzyloxy)phenyl)-6-isopropyl-4H-chromen-4-one

[0089] To a mixture of (E)-3-(3,4-bis(benzyloxy)phenyl)-l-(2-hydroxy-5-isopropylphenyl)prop-2- en-l-one

[0090] 0.18g, 0.376 mmol) in DMSO (3 mL) was added iodine (0.01g, 0.04 mmol), the reaction was heated at 130C. The completion of reaction was monitored by anal. HPLC. The reaction was cooled to rt and water was added, then extracted with EtOAc, washed with water, sat’d NaHCOs, and brine, and dried over Na2SO4. The solvent was removed to obtain the crude which was purified via silica gel flash chromatography to obtain the title compound. 'H NMR (400 MHz, CDCh) 5 8.05 (d, J=2.2 Hz, 1H), 7.56 (dd, J=8.2 Hz, 2.0 Hz, 1H), 7.49-7.31 (m, 13H), 7.02 (d, J=8.4 Hz, 1H), 6.68 (s, 1H), 5.25 (s, 4H), 3.06-3.00 (m, 1H), 1.30 (d, J=6.9 Hz, 6H) e. 2-(3,4-Dihydroxyphenyl)-6-isopropyl-4H-chromen-4-one

[0091] The mixture of 2-(3,4-bis(benzyloxy)phenyl)-6-isopropyl-4H-chromen-4-one (0.05g) and Pd / C (0.01g, 10%Pd on carbon) in EtOAcZEtOH (1 : 1, ImL) was degassed and stirred under EE balloon overnight. The completion of reaction was monitored by analytical HPLC. The reaction mixture was filtered through filter paper and the filtrate was concentrated to obtain the crude which was purified by prep. HPLC to obtain the title compound.1H NMR (400 MHz, CDCh and CD3OD) 5 7.99 (d, J=2.2 Hz, 1H), 7.61 (dd, J=8.2 Hz, 2.0 Hz, 1H), 7.02 (d, J=8.0 Hz, 1H), 7.45-7.40 (m, 2H), 6.92 (d, J=8.9 Hz, 1H), 6.70 (s, 1H), 3.08-3.00 (m, 1H), 1.30 (d, J=6.9 Hz, 6H)

[0092] Example 2. 6-(tert-Butyl)-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one a. 4-(tert-Butyl)phenyl acetate

[0093] The title compound was prepared following the same general protocol as described for Step a, Example 1, using 4-(tert-butyl)phenol. 'H NMR (400 MHz, CDCh) 57.42 (dd, J=6.7 Hz, 2.0Hz, 2H), 7.04 (dd, J=6.7 Hz, 2.0Hz, 2H), 2.31 (s, 3H), 1.34 (s, 9H). b. l-(5-(tert-Butyl)-2-hydroxyphenyl)ethenone

[0094] The title compound was prepared following the same general protocol as described for Step b, Example 1, using 4-(tert-butyl)phenyl acetate. *HNMR (400 MHz, CDCh) 5 12.05 (s, 1H), 7.61 (dd, J=8.7Hz, 2.4 Hz, 1H), 6.86 (d, J=8.7 Hz, 1H), 2.57 (s, 3H), 1.25 (s, 9H). c. (E)-3-(3,4-bis(Benzyloxy)phenyl)-l-(5-(tert-butyl)-2-hydroxyphenyl)prop-2-enone

[0095] The title compound was prepared following the same general protocol as described for Step c, Example 1, using l-(5-(tert-butyl)-2-hydroxyphenyl)ethenone. *H NMR (400 MHz, CDCh) 5 12.35 (s, 1H), 7.96 (d, J=2.6Hz, 1H), 7.80 (d, J=8.8Hz, 1H), 7.63-7.59 (m, 2H), 7.49- 7.31 (m, 12H), 7.16 (d, 8.4Hz, 1H), 8.95 (d, 8.4Hz, 1H), 5.26 (s, 2H), 5.23 (s, 2H), 1.35 (s, 9H). d. 2-(3,4-bis(Benzyloxy)phenyl)-6-(tert-butyl)-4H-chromen-4-one

[0096] The title compound was prepared following the same general protocol as described for Step d, Example 1, using (E)-3-(3,4-bis(benzyloxy)phenyl)-l-(5-(tert-butyl)-2-hydroxyphenyl)prop-2- enone.XH NMR (400 MHz, CDCl3) 58.05 (d, J=2.2 Hz, 1H), 7.64 (dd, J=8.2 Hz, 2.0 Hz, 1H), 7.39- 7.28 (m, 13H), 7.26 (d, J=8.4 Hz, 1H), 6.95 (s, 1H), 5.16 (s, 4H), 1.30 (s, 9H). e. 6-( / c / 7-Butyl)-2-(3.4-dihydroxyphenyl)-4H-chromen-4-one

[0097] The title compound was prepared following the same general protocol as described for Step e, Example 1, using 2-(3,4-bis(benzyloxy)phenyl)-6-(tert-butyl)-4H-chromen-4-one. 'H NMR (400 MHz, DMSO-d6) 57.97 (d, J=2.4 Hz, 1H), 7.91 (dd, J=8.2 Hz, 2.4 Hz, 1H), 7.67 (m, 2H), 6.91 (d, J=8.9 Hz, 1H), 6.75 (s, 1H), 1.35 (s, 9H). Example 3. 6-Cyclopentyl-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one (SR31133) a. (E)-3-(3,4-bis(Benzyloxy)phenyl)-l-(5-bromo-2-hydroxyphenyl)prop-2-en-l-one

[0098] To a mixture of l-(5-bromo-2-hydroxyphenyl)ethan-l-one (7.0 g, 32.55 mmol) and 3,4- bis(benzyloxy)benzaldehyde (10.36 g, 32.55mmol) in MeOH (80 mL) was added freshly made NaOMe in MeOH (120mL, 1.4M, 168.0 mmol) and the reaction was stirred at 90°C in an oil bath for 16h. The mixture was cooled to room temperature and poured into ice water, and the resulting mixture was adjusted to pH3-~4 by addition of 5N HC1. Filter the mixture to obtain the solid as the desired product with no further purification. ESI-MS (m / z): 514.8, 516.7 [M+l]+. b. 2-(3,4-bis(Benzyloxy)phenyl)-6-bromo-4H-chromen-4-one The title compound was prepared following the same general protocol as described for

[0099] Step d, Example 1, using (E)-3-(3,4-bis(benzyloxy)phenyl)-l-(5-bromo-2-hydroxyphenyl)prop- 2-en-l-one. ESI-MS (m / z): 512.8, 514.7 [M+l]+. c. 2-(3,4-bis(Benzyloxy)phenyl)-6-(cyclopent-l-en-l-yl)-4H-chromen-4-one

[0100] A mixture of 2-(3,4-bis(benzyloxy)phenyl)-6-bromo-4H-chromen-4-one (0.15 g, 0.292 mmol), 2-(cyclopent-l-en-l-yl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane (0.085g, 0.438 mmol), Pd(PPh3)4 (0.017g, 0.015 mmol) and K2CO3 (0.061g, 0.441 mmol) in dioxane (1 mL) and water (0.2mL) was stirred at 140°C overnight under Nitrogen. The mixture was added water and EtOAc, the mixture was stirred and separated, the aqueous phase was extracted with EtOAc twice, the combined organic phases were washed with brine and dried with Na2SO4, filtered through silica gel to get an oil, which was purified by silica gel column to obtain the title compound. ESI-MS (m / z): 501.0 [M+l]+. d. 6-Cyclopentyl-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one

[0101] The title compound was prepared following the same general protocol as described for Step e, Example 1, using 2-(3,4-bis(benzyloxy)phenyl)-6-(cyclopent-l-en-l-yl)-4H-chromen-4- one. ESI-MS (m / z): 322.9 [M+l]+.

[0102] Example 4. 6-Cyclohexyl-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one

[0103] 6-Cyclohexyl-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one

[0104] The title compound was prepared following the same general protocol as described for Step e, Example 1, using: 6-(cy cl ohex-l-en-l-yl)-2-(3,4-dihydroxyphenyl)-4H-chromen -4-one. 'HNMR (400 MHz, DMSO-d6) 5 7.84 (d, J=2.1 Hz, 1H), 7.71-7.63 (m, 2H), 7.45 -7.43 (m, 2H), 6.91 (d, J= 8.20, 1H), 6.74 (s, 1H), 2.67-2.64 (m, 1H), 1.86-1.51 (m 5H), 1.49-1.24 (m, 5H). Example 5. 2-(3,4-bis(Benzyloxy)phenyl)-6-(cyclohex-l-en-l-yl)-4H-chromen-4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-(cyclohex-l-en-l-yl)-4H-chromen-4-one

[0105] The title compound was prepared following the same general protocol as described for Step c, Example 3, using 2-(l-cyclohexenyl)-4,4,5,5-tetramethyl-l,3,2-dioxaborolane. ’H NMR (400 MHz, CDCh) 5 8.08 (d, J=2.3 Hz, 1H), 7.37 (dd, J=5.2 Hz, 2.4 Hz, 2H), 7.33 -7.17 (m, 12H), 6.94 (d, J= 8.28, 1H), 6.58 (s, 1H), 6.17 (q, J=2.32 Hz, 1H), 5.16 (s, 4H), 2.40-2.37 (m, 2H), 2.17-2.13 (m, 2H), 1.75-1.72 (m, 2H), 1.69-1.62 (m, 2H). b. 6-(Cyclohex-l-en-l-yl)-2-(3,4-dihydroxyphenyl)-4H-chromen-4-one

[0106] To the mixture of 2-(3,4-bis(benzyloxy)phenyl)-6-(cyclohex-l-en-l-yl)-4H-chromen-4- one (0.11 g, 0.22 mmol) in DCM (3 mL) at room temperature was added BBn (0.87 mL, IM in DCM, 0.87 mmol) dropwise. The completion of the reaction was monitored by anal. HPLC. The mixture was slowly poured into cold saturated NaHCCh, extracted with DCM, washed with brine and dried over Na2SO4. The solvent was removed to obtain the crude which was purified by prep-HPLC to obtain the title compound.1H NMR (400 MHz, DMSO-d6) 5 9.88 (br, 1H), 9.47 (br, 1H), 7.95 (s, 1H), 6.94 (dd, J= 8.20 Hz, 2.10 Hz, 1H), 7.68 (d, J=8.20, 1H), 7.46 (m, 2H), 6.92 (d, J=8.2Hz, 2.1Hz, 1H), 6.75 (s, 1H), 6.33 (t, J=4Hz, 1H), 2.44-2.43 (m, 2H), 2.24- 2.22 (m, 2H), 1.80-1.74 (m, 2H), 1.66-1.60 (m, 2H). Example 6. Preparation of 2-(3,4-Dihydroxyphenyl)-6-phenyl-4H-chromen-4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-phenyl-4H-chromen-4-one

[0107] The title compound was prepared following the same general protocol as described for Step c, Example 3, using phenylboronic acid. ESI-MS (m / z): 511.0 [M+l]+. b. 2-(3,4-Dihydroxyphenyl)-6-phenyl-4H-chromen-4-one

[0108] To the mixture of 2-(3,4-bis(benzyloxy)phenyl)-6-phenyl-4H-chromen-4-one (0.14 g, 0.292 mmol) in DCM (3 mL) at room temperature was added BBn (0.7 mL, IM in DCM, 0.7 mmol) dropwise. The completion of the reaction was monitored by anal. HPLC. The mixture was slowly poured into cold saturated NaHCCh, extracted with DCM, washed with brine and dried over Na2SO4. The solvent was removed to obtain the crude which was purified by prep- HPLC to obtain the title compound. ESI-MS (m / z): 511.0 [M+l]+.

[0109] Example 7. Preparation of 2-(3,4-dihydroxyphenyl)-6-(2-hydroxyphenyl)-4H-chromen- 4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-(2-hydroxyphenyl)-4H-chromen-4-one

[0110] The title compound was prepared following the same general protocol as described for Step c, Example 3, using (2-hydroxyphenyl)boronic acid. ESI-MS (m / z): 527.0 [M+l]+. b. 2-(3,4-Dihydroxyphenyl)-6-(2-hydroxyphenyl)-4H-chromen-4-one

[0111] The title compound was prepared following the same general protocol as described for Step b, Example 6, using 2-(3,4-bis(benzyloxy)phenyl)-6-(2-hydroxyphenyl)-4H-chromen-4- one. ESI-MS (m / z): 346.9 [M+l]+.

[0112] Example 8. 2-(3,4-Dihydroxyphenyl)-6-(3-hydroxyphenyl)-4H-chromen-4-one

[0113] 2-(3,4-bis(Benzyloxy)phenyl)-6-(3-hydroxyphenyl)-4H-chromen-4-one

[0114] The title compound was prepared following the same general protocol as described for Step c, Example 3, using (3-hydroxyphenyl)boronic acid. ESI-MS (m / z): 527.0 [M+l]+. b. 2-(3,4-Dihydroxyphenyl)-6-(3-hydroxyphenyl)-4H-chromen-4-one

[0115] The title compound was prepared following the same general protocol as described for Step b, Example 6, using 2-(3,4-bis(benzyloxy)phenyl)-6-(3-hydroxyphenyl)-4H-chromen-4- one. ESI-MS (m / z): 346.9 [M+l]+.

[0116] Example 9. 2-(3,4-Dihydroxyphenyl)-6-(3-methoxyphenyl)-4H-chromen-4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-(3-methoxyphenyl)-4H-chromen-4-one

[0117] The title compound was prepared following the same general protocol as described for Step c, Example 3, using (3-methoxyphenyl)boronic acid. 'H NMR (400 MHz, CDCh) 5 8.40 (d, J=2.2

[0118] Hz, 1H), 7.70 (dd, J=8.6 Hz, 2.2 Hz, 1H), 7.53 -7.28 (m, 15H), 7.22-7.17 (m, 2H), 6.92 (dd, J=8.1

[0119] Hz, 2.3 Hz, 2H), 5.23 (s, 2H), 5.20 (s, 2H), 3.85 (s, 3H). b. 2-(3,4-Dihydroxyphenyl)-6-(4-methoxyphenyl)-4H-chromen-4-one

[0120] The title compound was prepared following the same general protocol as described for Step e, Example 1, using 2-(3,4-bis(benzyloxy)phenyl)-6-(3-methoxyphenyl)-4H-chromen-4- one. ‘HNMR (400 MHz, DMSO-d6) 5 9.47 (s, 1H), 8.99 (s, 1H), 8.29 (d, J=2.3, 1H), 8.20 (dd, J=8.7Hz, 2.4Hz, 1H), 7.89 (d, J=8.7Hz, 1H), 7.56-7.44 (m, 3H), 7.40-7.33 (m, 2H), 7.06 (dd, J=7.4Hz, 1.8Hz, 1H), 3.92 (s, 3H). Example 10. 2-(3,4-Dihydroxyphenyl)-6-(4-methoxyphenyl)-4H-chromen-4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-(4-methoxyphenyl)-4H-chromen-4-one

[0121] The title compound was prepared following the same general protocol as described for Step c, Example 3, using (4-methoxyphenyl)boronic acid.JH NMR (400 MHz, CDCh) 5 8.48 (d, J=2.3 Hz, 1H), 7.70 (dd, J=8.2 Hz, 2.1 Hz, 1H), 7.59 -7.28 (m, 17H), 7.07-7.01 (m, 1H), 6.73 (s, 1H), 5.32 (s, 2H), 5.29 (s, 2H), 3.89 (s, 3H). c. 2-(3,4-Dihydroxyphenyl)-6-(4-methoxyphenyl)-4H-chromen-4-one

[0122] The title compound was prepared following the same general protocol as described for Step e, Example 1, using 2-(3,4-bis(benzyloxy)phenyl)-6-(4-methoxyphenyl)-4H-chromen-4- one.XH NMR (400 MHz, CD3OD) 5 8.31 (s, 1H), 8.07 (d, J=8.1 Hz, 1H), 7.77-7.66 (m, 3H), 7.50 (d, J= 8.20 Hz, 2H), 7.08 (d, J=8.1, 2H), 6.95 (d, J=8.1Hz, 1H), 6.79 (s, 1H), 3.87 (s, 3H).

[0123] Example 11. 2-(3,4-Dihydroxyphenyl)-6-(3-(trifluoromethyl)phenyl)-4H-chromen-4-one a. 2-(3,4-bis(Benzyloxy)phenyl)-6-(3-(trifluoromethyl)phenyl)-4H-chromen-4-one

[0124] The title compound was prepared following the same general protocol as described for Step c, Example 3, using (3-(trifluoromethyl)phenyl)boronic acid. 'H NMR (400 MHz, CDCh) 58.50 (d, J=2.3 Hz, 1H), 7.55 (dd, J=8.2 Hz, 2.1 Hz, 2H), 7.48 (dd, J=8.1Hz, 4.2Hz, 1Hz), 7.37 -7.17 (m, 15H), 7.95 (d, J=8.2Hz, 1H), 6.62 (s, 1H), 5.16 (d, J=1.9Hz, 4H). b. 2-(3,4-Dihydroxyphenyl)-6-(3-(trifluoromethyl)phenyl)-4H-chromen-4-one

[0125] The title compound was prepared following the same general protocol as described for Step e, Example 1, using 2-(3,4-bis(benzyloxy)phenyl)-6-(3-(trifluoromethyl)phenyl)-4H- chromen-4-one. 'H NMR (400 MHz, CD3OD) 5 8.39 (s, 1H), 8.15 (d, J=8.2 Hz, 1H), 8.00 (s, 2H), 7.84-7.58 (m, 3H), 7.58-7.48 (m, 2H), 6.96 (d, J=8.2Hz, 1H), 6.80 (s, 1H).

[0126] Biological Examples 12-16

[0127] Cells and Mice

[0128] Primary Erccl ^ mouse embryonic fibroblasts (MEFs) were isolated on embryonic day 12.5-13.5. In brief, mouse embryos were isolated from yolk sac followed by the complete removal of viscera, lung and heart if presented. Embryos were then minced into fine chunks, fed with MEFs medium, cultivated under 3% oxygen to reduce stresses. Cells were split at 1 :3 when reaching confluence. MEFs were grown at a 1 :1 ratio of Dulbecco’s Modification of Eagles Medium (supplemented with 4.5 g / L glucose and L-glutamine) and Ham’s F10 medium, supplemented with 10% fetal bovine serum, penicillin, streptomycin and non-essential amino acid. To induce oxidative stress-mediated DNA damage, Erccl ^ MEFs were switched to 20% oxygen for 3 passages.

[0129] Human IMR90 lung fibroblasts were obtained from American Type Culture Collection (ATCC) and cultured in EMEM medium with 10% FBS and pen / strep antibiotics. To induce senescence, cells were treated with 20 pM etoposide for 24 h, followed by 5 days in normal culture media. Human umbilical vein endothelial cells (HUVECs) were obtained from ATCC and cultured using Endothelial Cell Growth Media plus supplement [without vascular endothelial growth factor (VEGF)] and 1% pen / strep antibiotics (R&D Systems). The cells were experimentally treated at late passage 13 to 15.

[0130] Erccl+ / ~ and Erccl+ / Amice from C57BL / 6J and FVB / n backgrounds were crossed to generate Erccl~ / Amice to prevent potential strain-specific pathology. For drug treatment, 10- week-old Erccl~ / Amice were dosed with 20 mg / kg of FA Example 1 or vehicle by oral gavage three times per week up until 15 weeks of age. FA Example 1 was formulated in 2:98 of DMSO: vegetable oil for in vivo administration. Animal protocols used in this study were approved by University of Minnesota Institutional Animal Care and Use Committees.

[0131] Aged wild-type C57BL / 6J:FVB / NJ mice were generated by crossing C57BL / 6J and FVB / n inbred mice purchased from Jackson Laboratory (Stock No:000664 and 001800). Mice were left to age for 2 years before being enrolled into the late life intervention study. Once the cohort was of over two years, mice over 120 weeks were treated with 20 mg / kg of FAs Examples 1 and 3 or vehicle by oral gavage for five consecutive days and sacrificed two days after the last dose for tissue analysis. Fisetin and FAs were formulated in 2:98 of DMSO: vegetable oil for in vivo administration.. Animal protocols used in this study were approved by University of Minnesota Institutional Animal Care and Use Committees.

[0132] Health Evaluation of Erccl~ / AMice

[0133] Health assessment of Erccl~ / Amice was conducted twice per week to evaluate age- related symptoms, including body weight, tremor, forelimb grip strength, kyphosis, hindlimb paralysis, gait disorder, dystonia and ataxia. Kyphosis, body condition and coat condition were used to reflect general health conditions. Ataxia, dystonia, gait disorder and tremor were used as indicators of aging-related neurodegeneration. Muscle wasting was studies by monitoring hindlimb paralysis and forelimb grip strength. All aging symptoms were scored based on a scale of 0, 0.5 and 1, with the exception of dystonia that has a scale from 0 to 5. The sum of aging scores of each group was used to determine the overall aging conditions, with zero means no symptom presented. Statistical Analysis

[0134] Data were statistically analyzed by Graphpad Prism software. Two-tailed Student’s / -test was performed to determine differences between two groups, and one-way ANOVA with Tukey’s test was used for three groups. A value of p <0.05 was considered as statistically significant, shown as *p <0.05, **p <0.01, ***p < 0.001 and ****p<0.0001.

[0135] Example 12. Senolytic Activity

[0136] Senescence was evaluated based on SA-P-gal activity using C12FDG staining assay. Specifically, senescent ErccT^ MEFs were passaged for 3 times at 20% O2 to induce senescence then seeded at 3000 cells per well in black wall, clear bottom 96 well plates at least 16 hours prior to treatment. Following the addition of drugs, the MEFs were incubated for 48 hours at 20% O2. After removing the medium, cells were incubated in 100 nM Bafilomycin Al in culture medium for 60 minutes to induce lysosomal alkalinization, followed by incubation with 20 pM fluorogenic substrate C12FDG (Setareh Biotech, USA) for 2 hours and counterstaining with 2 pg / ml Hoechst 33342 (ThermoFisher, USA) for 15 minutes. Subsequently, cells were washed with PBS and fixed in 2% paraformaldehyde for 15 minutes. Finally, cells were imaged with 6 fields per well using a high content fluorescent image acquisition and analysis platform Cytation 1 (BioTek, VT, USA).

[0137] The senolytic activity of representative compounds was evaluated in senescent Ercd~ ~ mouse embryonic fibroblasts (MEFs) induced by oxidative stress. The selectivity was accessed using selectivity index (SI) which compares the EC50 values in non-senescent proliferating MEFs over senescent Ercd~ ~ MEFs. Results are provided in the following table.

[0138] Example 13: Physicochemical and Molecular Properties

[0139] The physicochemical and molecular properties of the compounds of Example 1 and Example 3 are summarized in the following Table. The compounds of Example 1 and Example 3 exhibited 2.9 and 27.5 times greater activity than fisetin, respectively. Increased lipophilicity was also observed in both compounds, with cLogP values of 3.87 and 4.49 for the compound of Example 1 and the compound of Example 3, respectively, compared to fisetin (1.84).

[0140] Topological polar surface area (tPSA) is an important indicator of a compound's ability to penetrate cells, and larger tPSA values are associated with lower permeability of the bloodbrain barrier (BBB) (van de Waterbeemd, H., et al., J Drug Target 6, 151-165 (1998)). For drugs targeting the central nervous system (CNS), tPSA values below 90 angstroms squared are generally necessary for penetration of the BBB (Pajouhesh, H. & Lenz, G.R. NeuroRXl, 541- 553 (2005)). Compared to fisetin, the compound of Example 1 and the compound of Example 3 exhibit lower tPSA values, which are less than 90 angstroms squared, suggesting that they may have increased potential as CNS drugs. Furthermore, both compounds have better computational molecular properties than fisetin in terms of drug score, drug-likeness, and theoretical toxicity risks (including mutagenicity, tumorigenicity, irritancy, and reproductive effects) calculated using Osiris Property Explorer. Table. Pharmacological and physicochemical properties.

[0141] Example 14. Plasma Stability Plasma (mouse and human) stability was evaluated by incubating 1 pM test compound with 100 pl serum. Aliquots were removed at 0, 5, 10, 20, 40, and 60 minutes and added to acetonitrile (5X v:v) to stop the reaction and precipitate the protein. At the end of the assay, the samples were centrifuged through a Millipore Multiscreen Solvinter 0.45 micron low binding Polytetrafluoroethylene (PTFE) hydrophilic filter plate and analyzed by LC-MS / MS. Data was log transformed and regression analysis was used to calculate half-life.

[0142] Example 15. In vitro Confirmation of the Senolytic Activity

[0143] To confirm the senolytic activity of the compound of Example 1 and the compound of Example 3, the compounds were tested in other senescent cell models, including human umbilical vein endothelial cells (HUVEC) and IMR90 fibroblasts. Senescent HUVECs were generated by serially passaging the cells until they reached a state of near growth arrest, typically at late passages 13 to 15. Human IMR90 fibroblast cells were induced senescent by genotoxic stress (etoposide). (Fuhnuarm-Stroissnigg, H., J Fw E p, 2019, Jun 28;(148);

[0144] PMID: 31305507). The compounds of Example 1 and Example 3 demonstrated a reduction in cellular senescence in senescent HUVEC cells, with ECso values of 6 pM and 8.73 pM, respectively. Additionally, these compounds showed a decrease in senescence in IMR90 cells, with ECso values of 20.98 pM and 3.895 pM, respectively.

[0145] Example 16. Senotherapeutic Potential of the FAs in vivo

[0146] Acute treatment reduced tissue senescence. As a starting point to confirm the in vivo therapeutic potential of the compound of Example 1 and the compound of Example 3, a naturally aging model of very old WT mice (27 months) was used in which fisetin would not be expected to be significantly effective (Yousefzadeh, M.J., el al. EBioMedicine 36, 18-28 (2018)). Instead of the 100 mg / kg dose used in fisetin studies, the old wild-type C57BL / 6J mice were acutely treated with only 20 mg / kg of body weight of vehicle, fisetin, the compound of Example 1 and the compound of Example 3 for five consecutive days. Mice were then sacrificed two days after the last administration. Molecular analysis of different tissues found that fisetin indeed showed marginal effect on senescence in these tissues whereas the compound of Example 1 and the compound of Example 3, especially the compound of Example 1, can significantly decrease the expression of multiple senescence and SASP markers in different tissues including kidney, brain, lung and spleen (Figs la-le).

[0147] Chronic treatment extended healthspan. The most effective fisetin analog of Example 1 in the acute mouse study was further selected to test in Erccl' progeria mouse model to determine its effect on tissue senescence and healthspan. Erccl'^ progeria mice were treated with 20 mg / kg of the compound of Example 1 for five weeks via oral administration (Fig 2a). Health assessments were conducted weekly to score age-related symptoms, including tremor, kyphosis, dystonia, ataxia, gait disorder, hindlimb paralysis and forelimb grip strength. Results demonstrated that the compound of Example 1 can suppress the composite score of aging symptoms, particularly tremor (Fig 2b). Gene expression analysis in tissues also showed a significant reduction or a trend towards significance in senescence and SASP factors in multiple tissues, including kidney, liver, lung, and brain (Fig 2c). There results further confirmed the promising senotherapeutic activity of the optimized compound of Example 1. Example 17. The following illustrate representative pharmaceutical dosage forms, containing a compound of formula I ('Compound X'), for therapeutic or prophylactic use in humans.

[0148] (i) Tablet 1 mg / tablet

[0149] Compound X= 100.0

[0150] Lactose 77.5

[0151] Povidone 15.0

[0152] Croscarmellose sodium 12.0

[0153] Microcrystalline cellulose 92.5

[0154] Magnesium stearate 3,0

[0155] 300.0

[0156] (ii) Tablet 2 mg / tablet

[0157] Compound X= 20.0

[0158] Microcrystalline cellulose 410.0

[0159] Starch 50.0

[0160] Sodium starch glycolate 15.0

[0161] Magnesium stearate 5,0

[0162] 500.0

[0163] (iii) Capsule mg / capsule

[0164] Compound X= 10.0

[0165] Colloidal silicon dioxide 1.5

[0166] Lactose 465.5

[0167] Pregelatinized starch 120.0

[0168] Magnesium stearate 3,0

[0169] 600.0 (iv) Injection 1 (1 mg / ml) mg / ml

[0170] Compound X= (free acid form) 1.0

[0171] Dibasic sodium phosphate 12.0

[0172] Monobasic sodium phosphate 0.7

[0173] Sodium chloride 4.5

[0174] 1.0 N Sodium hydroxide solution

[0175] (pH adjustment to 7.0-7.5) q.s.

[0176] Water for injection q.s. ad 1 mL

[0177] (v) Injection 2 (10 mg / ml ) mg / ml

[0178] Compound X= (free acid form) 10.0

[0179] Monobasic sodium phosphate 0.3

[0180] Dibasic sodium phosphate 1.1

[0181] Polyethylene glycol 400 200.0

[0182] 1.0 N Sodium hydroxide solution (pH adjustment to 7.0-7.5) q.s.

[0183] Water for injection q.s. ad 1 mL

[0184] (vi) Aerosol mg / can

[0185] Compound X= 20.0

[0186] Oleic acid 10.0

[0187] Trichloromonofluoromethane 5,000.0

[0188] Dichlorodifluoromethane 10,000.0

[0189] Di chlorotetrafluoroethane 5,000.0

[0190] The above formulations may be obtained by conventional procedures well known in the pharmaceutical art. All publications, patents, and patent documents are incorporated by reference herein, as though individually incorporated by reference. The invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention.

Claims

CLAIMSWhat is claimed is:1 A compound of formula I:or a salt thereof, wherein:R1is isopropyl, tert-butyl, 3 -methylpropyl, 1 -methylpropyl, (C3-C8)cycloalkyl, or phenyl, wherein any (C3-C8)cycloalkyl and phenyl is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro.2 The compound or salt of claim 1, wherein R1is isopropyl, tert-butyl, or 3 -methylpropyl.3 The compound or salt of claim 1, wherein R1is (C3-Ce)cycloalkyl that is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro.4 The compound or salt of claim 1, wherein R1is phenyl that is optionally substituted with one or more groups independently selected from the group consisting of, hydroxy, methoxy and methyl, which methoxy and methyl are optionally substituted with one or more fluoro.5 The compound or salt of claim 1, wherein R1is isopropyl, tert-butyl, 3 -methylpropyl, cyclopentyl, cyclohexyl, 1 -cyclohexenyl, phenyl, 2-hydroxyphenyl, 3 -hydroxyphenyl, 3- methoxyphenyl, 4-methoxyphenyl, or 3 -trifluoromethylphenyl.

6. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

7. A method for reducing the expression of one or more senescence markers in a cell comprising, contacting the cell with a compound of formula (I) as described in any one of claims 1- 5 or a salt thereof.8 The method of claim 7, wherein the contacting occurs in vitro.9 The method of claim 7, wherein the contacting occurs in an animal.10 A method for reducing senescence in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.11 A method for extending the healthspan or lifespan of an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.12 A method for treating age-related disease in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.13 A method for increasing senotherapeutic activity in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.14 A method for treating a disease or condition in an animal selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bonemarrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

15. A method for improving response to chemotherapy in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

16. A method for improving response to radiation therapy in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

17. A method for improving recovery of a cancer survivor, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the cancer survivor.

18. A method for improving cognition in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

19. A method for reducing the negative effects of tobacco use (past or present) in an animal, comprising a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

20. A method for reducing one or more symptoms associated with long-haul COVID-19 in an animal, comprising administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

21. A method comprising contacting an organ with a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to improve the characteristics of the organ for transplantation.

22. A method for improving bone healing in an animal comprising, administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

23. A method for repairing damaged muscle in an animal comprising, administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

24. A method for improving tissue homeostasis comprising, contacting tissue with a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof.

25. A method for improving immune response to a vaccine in an animal comprising, administering a compound of formula (I) as described in any one of claims 1-5, or a pharmaceutically acceptable salt thereof to the animal.

26. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for use in medical therapy.

27. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for reducing senescence, extending healthspan, treating an age-related disease, or increasing senotherapeutic activity.

28. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing senescence, extending healthspan, treating an age-related disease, or increasing senotherapeutic activity in an animal.

29. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for treating a disease or condition selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bone marrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection.

30. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving response to chemotherapy.

31. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving response to radiation therapy.

32. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving recovery of a cancer survivor.

33. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving cognition.

34. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for reducing the negative effects of tobacco use.

35. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for reducing one or more symptoms associated with long-haul COVID-19.

36. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving the characteristics of an organ for transplantation.

37. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving bone healing.

38. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for repairing damaged muscle.

39. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving tissue homeostasis.

40. A compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof for improving immune response to a vaccine.

41. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for treating a disease or condition selected from the group consisting of diabetes, obesity, age-related lipodystrophy, cardiac dysfunction including atrial fibrillation, vascular hyporeactivity, vascular calcification, AV fistulae, frailty, sarcopenia, sequellae of bone marrow transplantation, sequellae of organ transplantation, myeloma, MGUS, Alzheimer’s disease, Parkinson’s disease, cancer survivors, ALS, anxiety, renal dysfunction, osteoporosis, osteoarthritis, rheumatoid arthritis, COPD, idiopathic pulmonary fibrosis, hyperoxic lung damage, hepatic steatosis, liver cirrhosis, primary biliary cirrhosis, liver, kidney and lung fibrosis, progerias, critical illness myopathy, pre-eclampsia, uterine fibrosis, ovarian involution, cataracts, macular degeneration, glaucoma, prostatic hypertrophy, skin disorders, scleroderma, stem cell activation, progenitor cell dysfunction, improving immune function including an immune response to vaccines, COVID-19, a viral infection, and a bacterial infection in an animal.

42. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving response to chemotherapy in an animal.

43. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving response to radiation therapy in an animal.

44. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving recovery of a cancer survivor.

45. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving cognition in an animal.

46. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing the negative effects of tobacco use (past or present) in an animal.

47. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for reducing one or more symptoms associated with long-haul COVID-19 in an animal.

48. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving the characteristics of an organ for transplantation.

49. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving bone healing in an animal.

50. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for repairing damaged muscle in an animal.

51. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving tissue homeostasis in an animal.

52. The use of a compound of formula I as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof to prepare a medicament for improving immune response to a vaccine in an animal.

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