Amino acid salts of nicotinic acid ribosides as anti-aging agents

Abstract

The present invention relates to amino acid salts of nicotinic acid riboside of Formula I and compositions thereof, which are useful in the treatment of conditions and diseases associated with NAD+deficiency: wherein M 1 , R 1 , R 2 and R 3 as described herein.

Description

[0001] Related applications

[0002] This application claims priority and benefit to U.S. Provisional Application No. 62 / 675,065, filed May 22, 2018, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This invention relates to inorganic salts and compositions thereof of nicotinic acid nucleoside, which can be used to treat age-related conditions and diseases. Background Technology

[0004] Aging is the result of complex interactions involving biological, physical, and biochemical processes that lead to cellular and organ dysfunction, manifesting as a variety of diseases and other outcomes. For example, female fertility is highly sensitive to the effects of aging. For instance, the U.S. Centers for Disease Control and Prevention (CDC) reports that among women around 35 years of age, the percentage of pregnancies and deliveries associated with assisted reproductive technology (ART) steadily declines from approximately 25% in ART cycles, resulting in a decrease in singleton live births to 14% by age 40 (CDC, American Academy of Reproductive Medicine, Assisted Reproductive Technology Association. 2011 Assisted Reproductive Technology National Summary Report. Atlanta (GA): US Dept of Health and Human Services; 2013). This trend increases significantly beyond age 40, with the CDC reporting a very low success rate for women over 44. In this group, the percentage of live births and singleton live births declines to approximately 1%. Age is generally considered the most significant factor influencing the chances of a live birth when a woman uses her own eggs (oocytes).

[0005] It should be understood that the deterioration of oocyte quality due to aging is a fundamental factor in declining fertility. For example, it has been reported that oocytes in older women are prone to chromosomal division abnormalities, exhibiting decreased mitochondrial quality, low ATP production, increased oxidative stress, and reduced antioxidant levels (Nelson SM, Telfer EE, Anderson RA. The aging ovary and uterus: new biological insights. Hum Reprod Update. 2013; 19:67-83.; Wilding M. Potential long-term risks associated with maternal aging (the role of the mitochondria). Fertil Steril. 2015; 103:1397-401; 3. Meldrum DR, Casper RF, Diez-Juan A, Simon C, Domar AD, Frydman R. Aging and the environment affect gamete and embryo potential: can we intervene? Fertil Steril. 2016; 105:548-59).

[0006] For all the reasons mentioned above, oocytes represent an excellent target tissue for evaluating therapeutic modalities that are expected to have an impact on the aging process, and in addition, offer the prospect of addressing age-related infertility.

[0007] One possible therapeutic modality for treating aging includes increasing NAD+. + The therapeutic level of NAD. + It is an important component of cellular processes essential for supporting a variety of metabolic functions. NAD + The classic role of NAD+ is as a coenzyme, catalyzing cellular redox reactions in many fundamental metabolic processes, such as glycolysis, fatty acid β-oxidation, or the tricarboxylic acid cycle, to be reduced to NADH. In addition to these roles, NAD+... + Also used as a source of NAD. + Substrates of these enzymes (such as poly-ADP-ribose polymerase (PARP), sirtuin, and CD38 / 157 extracellular enzymes) play a crucial role. These enzymes are known to consume NAD+. + Enzymes mediate many basic cellular processes.

[0008] There are five main synthetic NADs +Precursors and intermediates: tryptophan, nicotinamide, nicotinic acid (NA), nicotinamide riboside (NR), and nicotinamide mononucleotide (NMN). NAD + The amino acid tryptophan can be converted into nicotinic acid mononucleotide (NaMN) through multiple enzymatic steps for de novo synthesis. NaMN is then converted into nicotinic acid dinucleotide (NaAD) via NMN / NaMN adenylate transferase (NMNAT). + ), and then through NAD + synthase amidation to NAD + .

[0009] In mammals, NAD + The primary biosynthetic pathway for nicotinamide is the salvage pathway. Nicotinamide is converted to the crucial NAD+ intermediate NMN by the rate-limiting enzyme nicotinamide phosphoribosyltransferase (NAMPT) in this pathway. NMN is then converted to NAD+ by NAMPT. + NAMPT plays a crucial role in regulating cellular NAD+ levels. On the other hand, niacin is converted to NaMN via nicotinic acid phosphoribosyltransferase (NPT). NR needs to be converted to NMN via phosphorylated NR16 nicotinamide ribokinases NMRK1 and NMRK2 (also known as NRK1 and NRK2). Maintaining sufficient NAD+ levels is essential. + Biosynthesis is essential for cell survival and function. Deviations in NAD+... + Steady state not only significantly affects the NAD required for redox reactions + / NADH library, and NAD can affect key cellular functions + The activity of NAD+-dependent enzymes. Currently, NAD+ levels at the cellular, tissue / organ, and organismal levels... + It is widely accepted that NAD+ levels decline during aging. + The activity of this enzyme is affected by NAD. + The decline has a diminishing effect, leading to various age-related effects.

[0010] Nicotinamide adenine dinucleotide (NAD) is an enzyme cofactor essential for the function of several enzymes involved in reductive-oxidative reactions and energy metabolism. (Katrina L. Bogan and Charles Brenner, Nicotinic Acid, Nicotinamide and Nicotinamide Riboside: A Molecular Evaluation of NAD) + Precursor Vitamins in Nutritions, 28, Annual Review of Nutrition 115 (2008). NAD +It acts as an electron carrier in the energy metabolism of amino acids, fatty acids, and carbohydrates (Bogan & Brenner, 2008). NAD + It is crucial for redox reactions and as a substrate for PARP (polyadenylated diphosphate ribose polymerase) and sirtuin (SIRT1 to SIRT7) signaling, playing a vital role in DNA repair, energy metabolism, cell survival, and the regulation of circadian rhythms (Bronkowski, MS & Sinclair, D., Nat. Rev. Mole. Cell. Bio., 17, 679-690, 2016). Enhancing NAD+... + Concentrations of NAD+ delay aging in yeast, flies, and mice (Mouchiroud et al., Cell 154, 464-471, 2014). It has also recently been shown that NAD+... + Directly regulating protein-protein interactions, and modulating these interactions, can prevent cancer and radiation exposure and has a direct impact on aging (Li et al., Science 355, 1312-1317, 2017). Therefore, mounting evidence supports the idea that NAD+ can be used to... + Intervention with intermediates (such as NMN and NR) can restore available NAD. + To enhance the system and mitigate age-related physiological decline.

[0011] Although NAD + NAD can be synthesized de novo from the amino acid tryptophan, but this process does not occur in all tissues. Most cells rely on salvage pathways (as described above) to regenerate NAD from other intracellular intermediates primarily available through dietary sources. +(Christopher R. Martens, et al., Nat. Commun. 9, 1286, (2018) and Bogan, KL & Brenner, C., Annu. Rev. Nutr. 28, 115-130, (2008)). Other NAD precursors (such as nicotinic acid and nicotinamide) can also be administered to enhance NAD cell bioavailability. However, clinically relevant levels of niacin at therapeutic doses have been associated with undesirable hot flashes (MacKay, D., Hathcock, J. & Guarneri, E., Nutr. Rev. 70, 357-366 (2012)), and despite elevated NAD concentrations, nicotinamide does not reliably activate (and may even inhibit) sirtuin (Bitterman, KJ, et al., J. Biol. Chem. 277, 45099-45107 (2002); Guan, X., et al., PLoS One. 9, e107729 (2014); and Trammell, SA, et al., Nat. Commun. 7, 12948 (2016)). Therefore, administration of niacin or nicotinamide is unlikely to be widely used for maintaining health and function with age.

[0012] Compared with nicotinic acid and nicotinamide, in animal models, administration of NAD+... + Metabolites such as nicotinamide mononucleotide (NMN) or nicotinamide ribose (NR) appear to increase NAD. + It increases NAD levels and improves various physiological functions (Yoshino, J. et al., Cell Metab. 14, 528-536 (2011); Mills, KF et al., Cell Metab. 24, 795-806 (2016); and Frederick, DW et al., Cell Metab. 24, 269-282 (2016)). At least one of these metabolites has been reported to be well-tolerated in humans, leading to increased NAD levels and improved physiological functions, although further research is needed to confirm the findings of this exploratory study (Christopher R. Martens et al., Nat. Commun. 9, 1286, (2018)). Furthermore, recent studies have shown that a single dose of NR stimulates NAD levels in blood cells of healthy individuals in a dose-dependent manner. + Metabolism (Trammell, SA et al., Nat. Commun. 7, 12948 (2016)) demonstrates the limitations of such metabolites. However, many known NAD+ metabolites are unstable in a variety of physiological environments and therefore cannot be used as feasible drugs for patients who require such metabolites to increase NAD+ levels in their patients.

[0013] Considering NAD + NAD plays a central role in key cellular and physiological pathways; development of NAD can improve disease states and / or aging processes. + New stabilizers with improved properties at different levels are essential for improving human conditions. Summary of the Invention

[0014] This article provides amino acid salts of NaR that surprisingly increase cellular NAD. + level.

[0015] The first aspect of this application relates to salts of formula (I): and their enantiomers, stereoisomers, and tautomers.

[0016]

[0017] in

[0018] M 1 It is an amphoteric amino acid;

[0019] R 1 R 2 and R 3 Independently H, C1–C6 alkyl, C1–C6 haloalkyl, (C0–C3 alkylene)C(O)C1–C6 alkyl, C(O)R a -C(O)OR a -C(O)NR a R b Or -[CH2-CH2-O] k -R a ,

[0020] Or R 1 and R 2 Or R 2 and R 3 Together with the atoms to which it is attached, it forms a 5-membered heterocycle optionally substituted with one or more of the following substituents: C1–C6 alkyl, C2–C6 alkenyl, C2–C6 ynyl, (C0–C3 alkylene)C3–C8 cycloalkyl, (C0–C3 alkylene) heterocycloalkyl, (C0–C3 alkylene)C6–C 14 Aryl or (C0–C3 alkylene) heteroaryl;

[0021] R a and R b Each time it appears, it is independently H or C1–C6 alkyl, wherein the alkyl group is optionally substituted by one or more substituents selected from: (C0–C3 alkylene)C3–C8 cycloalkyl, (C0–C3 alkylene) heterocycloalkyl, (C0–C3 alkylene)C6–C14 aryl or (C0–C3 alkylene) heteroaryl; and

[0022] k is an integer from 1 to 8.

[0023] Another aspect of this disclosure relates to a pharmaceutical composition comprising a salt of Formula I or a pharmaceutically acceptable salt thereof associated with a pharmaceutically acceptable carrier.

[0024] Another aspect of this application relates to a method for treating or preventing age-related conditions, the method comprising administering to a subject in need an effective amount of a salt of formula (I) or an enantiomer, stereoisomer or tautomer thereof.

[0025] Another aspect of this application relates to a method for treating or preventing infertility, the method comprising administering to a subject in need an effective amount of a salt of formula (I) or an enantiomer, stereoisomer or tautomer thereof.

[0026] Another aspect of this application relates to a salt of formula (I) or its enantiomers, stereoisomers or tautomers, for use in treating age-related conditions.

[0027] Another aspect of this application relates to a salt of formula (I) or its enantiomer, stereoisomer or tautomer, for use in treating infertility.

[0028] Another aspect of this application relates to the use of a salt of formula (I) or its enantiomers, stereoisomers or tautomers in the manufacture of a medicament for treating age-related conditions.

[0029] Another aspect of this application relates to the use of a salt of formula (I) or its enantiomers, stereoisomers or tautomers in the manufacture of a medicament for treating infertility.

[0030] Another aspect of this disclosure relates to a method for improving oocyte quality and maturation, the method comprising administering a therapeutically effective amount of a salt of formula I to a subject in need of such treatment.

[0031] Another aspect of this disclosure relates to the use of a salt of formula (I) or its enantiomers, stereoisomers or tautomers in the manufacture of a medicament for treating age-related conditions.

[0032] In another aspect, the present invention includes ex vivo treatment of oocytes with a salt of formula (I) prior to implantation into a subject for the treatment of age-related infertility.

[0033] In another aspect, the present invention includes ex vivo treatment of the blastocyst with a salt of formula (I) prior to implantation into a subject for the treatment of age-related infertility.

[0034] In another aspect, the present invention includes ex vivo treatment of oocytes with a salt of formula (I) prior to implantation into a subject for the treatment of infertility.

[0035] In another aspect, the present invention includes ex vivo treatment of the blastocyst with a salt of formula (I) prior to implantation into a subject for the treatment of infertility.

[0036] On the other hand, a salt of formula (I) is provided as a component in a solution for in vitro cell treatment for the treatment of age-related conditions. In some embodiments, the age-related condition is age-related infertility. In other aspects, a salt of formula (I) is provided as a component in a solution for in vitro cell treatment for the treatment of infertility.

[0037] Another aspect of this disclosure relates to a method for preparing a salt of formula (I), the method comprising contacting a nicotinic acid mononucleotide derivative of formula II with an alkali metal hydroxide under suitable conditions for effectively producing a salt of formula I.

[0038] This disclosure also relates to a method for accelerating recovery from a disease or condition. The method includes administering to a subject in need an effective amount of a salt of formula (I) in combination with a prescribed treatment for the disease.

[0039] On the other hand, this disclosure relates to a cell culture medium for in vitro fertilization, comprising: one or more salts of formula (I) and a culture agent. Detailed Implementation

[0040] This application relates to salts and compositions capable of treating or preventing age-related conditions. The application is characterized by a method of treating, preventing, or alleviating age-related diseases or conditions by administering a therapeutically effective amount of a salt of formula (I) or its enantiomers, stereoisomers, or tautomers to a patient in need. The method of this application can be used to treat a variety of diseases and conditions by preventing or alleviating aging and cellular regeneration processes (including but not limited to infertility and cellular degeneration).

[0041] The salts of formula (I) are potent and effective at clinically achievable doses; stable in a variety of potential dosage forms; have acceptable solubility and pH values; are crystalline; have reduced hygroscopicity; and are easy to handle—all of which are consistent with the development, manufacture, and use of the drug. Furthermore, the salts disclosed herein provide enhanced biological activity to increase cellular NAD+. + The level increases stability and makes the pH more physiologically acceptable.

[0042] The first aspect of this disclosure relates to a salt of formula I.

[0043]

[0044] Where M1 R 1 R 2 and R 3 As described in this article.

[0045] The article “a / an” is used in this disclosure to refer to the grammatical object of an article that is one or more (i.e., at least one). For example, “a / an element” means one or more elements.

[0046] Unless otherwise indicated, the term “and / or” is used in this disclosure to mean “and” or “or”.

[0047] The term "optionally substituted" should be understood to mean that a given chemical moiety (e.g., alkyl) may (but is not required to) be bonded to other substituents (e.g., heteroatoms). For example, an optionally substituted alkyl group may be a fully saturated alkyl chain (i.e., a pure hydrocarbon). Alternatively, the same optionally substituted alkyl group may have substituents other than hydrogen. For example, it may be bonded to a halogen atom, a hydroxyl group, or any other substituent described herein at any point along the chain. Thus, the term "optionally substituted" means that a given chemical moiety has the potential to contain other functional groups, but does not necessarily have any other functional groups. Suitable substituents used in the optional substitution of the stated group include, but are not limited to, halogens, oxo groups, -OH, -CN, -COOH, -CH2CN, -O-(C1-C6)alkyl, (C1-C6)alkyl, C1-C6 alkoxy, (C1-C6)haloalkyl, C1-C6 haloalkoxy, -O-(C2-C6)alkenyl, -O-(C2-C6)ynyl, (C2-C6)alkenyl, (C2-C6)ynyl, -OH, and -OP(O)(OH). 2. -OC(O)(C1-C6)alkyl, -C(O)(C1-C6)alkyl, -OC(O)O(C1-C6)alkyl, -NH2, -NH((C1-C6)alkyl), -N((C1-C6)alkyl)2, -NHC(O)(C1-C6)alkyl, -C(O)NH(C1-C6)alkyl, -S(O)2(C1-C6)alkyl, -S(O)NH(C1-C6)alkyl, and S(O)N((C1-C6)alkyl)2. The substituent itself may be optionally substituted. As used herein, "optionally substituted" also means substituted or unsubstituted, as defined below.

[0048] As used herein, the term “substituted” means that a specified group or portion has one or more suitable substituents, wherein the substituents may be attached to the specified group or portion at one or more positions. For example, an aryl group substituted with a cycloalkyl group may mean that the cycloalkyl group is attached to an atom of the aryl group by a bond or by fusion with the aryl group and sharing two or more common atoms.

[0049] As used herein, the term “unsubstituted” means that the specified group does not contain substituents.

[0050] Unless otherwise specifically defined, the term "aryl" refers to a cyclic aromatic hydrocarbon group having one to three aromatic rings, including monocyclic or bicyclic groups such as phenyl, biphenyl, or naphthyl. In the case of two aromatic rings (bicyclic, etc.), the aromatic rings of the aryl group may be connected at a single point (e.g., biphenyl) or fused (e.g., naphthyl). The aryl group may optionally be substituted with one or more substituents, for example, one to five substituents, at any connection point. Exemplary substituents include, but are not limited to: -H, halogen, -O-(C1-C6)alkyl, (C1-C6)alkyl, -O-(C2-C6)alkenyl, -O-(C2-C6)ynyl, (C2-C6)alkenyl, (C2-C6)ynyl, -OH, -OP(O)(OH)2, -OC(O)(C1-C6)alkyl, -C(O)(C1-C6)alkyl, -OC(O)O(C1-C6)alkyl, NH2, NH((C1-C6)alkyl), N((C1-C6)alkyl)2, -S(O)2-(C1-C6)alkyl, -S(O)NH(C1-C6)alkyl, and S(O)N((C1-C6)alkyl)2. The substituent itself may be optionally substituted. Furthermore, when containing two fused rings, the aryl group as defined herein may have unsaturated or partially saturated rings fused to a fully saturated ring. Exemplary ring systems of these aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl, anthracene, phenalenyl, phenanthrene, indanyl, indenyl, tetrahydronaphthyl, tetrahydrobenzoannulenyl, etc.

[0051] Unless otherwise specifically defined, "heteroaryl" means a monovalent monocyclic or polycyclic aromatic group with 5 to 24 ring atoms, containing one or more cyclic heteroatoms selected from N, O, or S, with the remaining ring atoms being C. As defined herein, "heteroaryl" also means a bicyclic heteroaryl group, wherein the heteroatoms are selected from N, O, or S. The aromatic group may optionally be independently substituted by one or more substituents as described herein.Examples include, but are not limited to, furanyl, thiophene, pyrrolyl, pyridyl, pyrazolyl, pyrimidinyl, imidazoleyl, isoxazolyl, oxazolyl, oxadiazolyl, pyrazinyl, indoleyl, thiophene-2-yl, quinolinyl, benzopyranyl, isothiazolyl, thiazolyl, thiadiazole, indazole, benzimidazolyl, thieno[3,2-b]thiophene, triazolyl, triazinyl, imidazo[1,2-b]pyrazolyl, furan[2,3-c]pyridyl, imidazo[1,2-a]pyridyl, indazoleyl, pyrrolo[2,3-c]pyridyl, pyrrolo[3,2-c]pyridyl, pyrazolo[3,4-c]pyridyl, thieno[3,2-c]pyridyl, thieno[2, [3-c]pyridyl, thieno[2,3-b]pyridyl, benzothiazolyl, indole, indolinyl, indolinone, dihydrobenzothiophene, dihydrobenzofuran, benzofuran, benzodihydropyranyl, thiobenzodihydropyranyl, tetrahydroquinolinyl, dihydrobenzothiazine, dihydrobenzoxanyl, quinolinyl, isoquinolinyl, 1,6-naphthidyl, benzo[de]isoquinolinyl, pyrido[4,3-b][1,6]naphthidyl, thieno[2,3-b]pyrazinyl, quinazolinyl, tetrazo[1,5-a]pyridyl, [1,2,4]triazo[4,3-a]pyridyl, isoindole pyrrolo[2,3-b]pyridyl, pyrrolo[3,4-b]pyridyl, pyrrolo[3,2-b]pyridyl, imidazo[5,4-b]pyridyl, pyrrolo[1,2-a]pyrimidinyl, tetrahydropyrrolo[1,2-a]pyrimidinyl, dibenzo[b,d]thiophene, pyridin-2-one, furano[3,2-c]pyridyl, furano[2,3-c]pyridyl, 1H-pyrido[3,4-b][1,4]thiazinyl, benzoxazolyl, benzoisoxazolyl, furano[2,3-b]pyridyl, benzothiophene, 1,5-naphthidyl, furano[3,2-b]pyridine, [1,2,4]triazol[1,5-a] [Pyridyl, benzo[1,2,3]triazolyl, imidazo[1,2-a]pyrimidinyl, [1,2,4]triazol[4,3-b]pyridazinyl, benzo[c][1,2,5]thiadiazolyl, benzo[c][1,2,5]oxadiazole, 1,3-dihydro-2H-benzo[d]imidazol-2-one, 3,4-dihydro-2H-pyrazolo[1,5-b][1,2]oxazinyl, 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridyl, thiazo[5,4-d]thiazolyl, imidazo[2,1-b][1,3,4]thiadiazolyl, thieno[2,3-b]pyrroleyl, 3H-indolyl and their derivatives. Furthermore, when containing two fused rings, the aryl group as defined herein may have unsaturated or partially saturated rings fused with a fully saturated ring.Exemplary ring systems of these heteroaryl groups include indololinyl, indololinone, dihydrobenzothiophene, dihydrobenzofuran, benzodihydropyranyl, thiobenzodihydropyranyl, tetrahydroquinolinyl, dihydrobenzothiazine, 3,4-dihydro-1H-isoquinolinyl, 2,3-dihydrobenzofuran, indololinyl, indolyl, and dihydrobenzoxyl.

[0052] Halogens, or "halogenated groups," refer to fluorine, chlorine, bromine, or iodine.

[0053] Alkyl refers to a straight-chain or branched saturated hydrocarbon containing 1 to 12 carbon atoms. Examples of (C1-C6) alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, neopentyl, and isohexyl.

[0054] "Alkoxy" refers to a straight-chain or branched saturated hydrocarbon containing 1-12 carbon atoms, i.e., -O (alkyl). Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, tert-butoxy, or pentoxy.

[0055] "Alkenyl" refers to a straight-chain or branched unsaturated hydrocarbon containing 2 to 12 carbon atoms. An alkenyl group contains at least one double bond in its chain. The double bond in an alkenyl group may be unbonded or bonded to another unsaturated group. Examples of alkenyl groups include vinyl, propenyl, n-butenyl, isobutenyl, pentenyl, or hexenyl. Alkenyl groups can be unsubstituted or substituted. As defined herein, alkenyl groups can be straight-chain or branched.

[0056] "Alynyl" refers to a straight-chain or branched unsaturated hydrocarbon containing 2 to 12 carbon atoms. An alkynyl group contains at least one triple bond in its chain. Examples of alynyl groups include ethynyl, propynyl, n-butynyl, isobutynyl, pentyynyl, or hexynyl. Alynyl groups can be unsubstituted or substituted.

[0057] The term "alkylene" or "alkylenyl" refers to a divalent alkyl group. Any of the monovalent alkyl groups described above can become an alkylene group by abstracting a second hydrogen atom from the alkyl group. As defined herein, alkylene groups can also be C1-C6 alkylene groups. Alkylene groups can further be C1-C4 alkylene groups. Typical alkylene groups include, but are not limited to: -CH2-, -CH(CH3)-, -C(CH3)2-, -CH2CH2-, -CH2CH(CH3)-, -CH2C(CH3)2-, -CH2CH2CH2-, -CH2CH2CH2CH2-, etc.

[0058] "Cycloalkyl" refers to compounds containing 3-18 carbon atoms (e.g., C3-C4). 10Cycloalkyl is a monocyclic or polycyclic saturated carbocyclic ring (e.g., fused ring, bridging ring, or spirocyclic). Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, norbornyl, norbornyl, bicyclo[2.2.2]octyl, or bicyclo[2.2.2]octenyl.

[0059] "Heterocyclic" or "heterocyclic alkyl" refers to a monocyclic or polycyclic ring (e.g., fused ring, bridging ring, or spirocyclic) containing a carbon atom and a heteroatom derived from oxygen, nitrogen, or sulfur, and wherein there is no delocalized π electron (aromaticity) shared between the ring carbon atoms or heteroatoms. Heterocyclic alkyl groups can be 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12-membered rings. The heterocyclic alkyl ring structure can be substituted by one or more substituents. The substituents themselves can be optionally substituted. Examples of heterocyclic groups include, but are not limited to, oxacyclobutane, azetadinyl, tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, oxazolinyl, oxazolinyl, thiazolinyl, thiazolinyl, pyranyl, thiopyranyl, tetrahydropyranyl, dioxalinyl, piperidinyl, morpholinyl, thiomorpholinyl, thiomorpholinyl S-oxide, thiomorpholinyl S-dioxide, piperazine, and azetadinyl. basic, oxygen basalt, diazoxide The terms include tropanyl, oxazolidinone, and homotropanyl. According to this application, a 3- to 10-membered heterocyclic group refers to a saturated or partially saturated non-aromatic ring structure containing between 3 and 10 atoms, wherein at least one heteroatom selected from the group N, O, or S is present.

[0060] The term "hydroxyalkyl" means an alkyl group as defined above, wherein the alkyl group is substituted with one or more OH groups. Examples of hydroxyalkyl groups include HO-CH2-, HO-CH2-CH2-, and CH3-CH(OH)-.

[0061] As used herein, the term "haloalkyl" refers to an alkyl group as defined herein that has been substituted with one or more halogens. Examples of haloalkyl groups include, but are not limited to, trifluoromethyl, difluoromethyl, pentafluoroethyl, trichloromethyl, etc.

[0062] As used herein, the term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens as defined herein. Examples of alkyl halogens include, but are not limited to, trifluoromethoxy, difluoromethoxy, pentafluoroethoxy, trichloromethoxy, etc.

[0063] As used herein, the term "cyano" refers to a substituent having a carbon atom connected to a nitrogen atom via a triple bond (i.e., C≡N).

[0064] As used in this article, the term "amine" refers to primary amines (R-NH2, R≠H) and secondary amines (R-NH2, R≠H). a -NH,R b ≠H) and tertiary amines (R) a -N, R≠H). A substituted amine is an amine in which at least one hydrogen atom has been replaced by a substituent.

[0065] As used herein, the term "amino" means a substituent containing at least one nitrogen atom. Specifically, the term "amino" includes NH2, -NH (alkyl) or alkylamino, -N (alkyl)2 or dialkylamino, amide-, urea-, urea and sulfonamide substituents.

[0066] As used in this article, the term "oxo" refers to the "=O" group.

[0067] The term "isomer" refers to salts and / or compounds that have the same composition and molecular weight but different physical and / or chemical properties. Structural differences can be in their configuration (geometric isomers) or their ability to rotate the plane of polarization (stereoisomers). Regarding stereoisomers, the salt of formula (I) may have one or more asymmetric carbon atoms and may exist as racemates, racemic mixtures, and as individual enantiomers or diastereomers.

[0068] This disclosure also includes pharmaceutical compositions comprising an effective amount of the disclosed salt and a pharmaceutically acceptable carrier. Representative “pharmaceutically acceptable salts” include, for example, water-soluble and water-insoluble salts such as acetates, aniline (4,4-diaminostilbene-2,2-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, tartrate, borate, bromide, butyrate, calcium salt, calcium edetate, dextrorotatory camphor sulfonate, carbonate, chloride, citrate, clavulanate, dihydrochloride, edetate, ethanedisulfonate, propionate, dodecyl sulfate, ethanesulfonate, fumarate, fiunarate, gluconate, gluconate, glutamate, glycolyllarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, and salts. Salts, hydroxynaphthylcarboxylate, iodides, isothionate, lactate, lacturonate, laurate, magnesium salts, malate, maleate, mandelate, methanesulfonate, methyl bromide, methyl nitrate, methyl sulfate, mucilage, naphthalenesulfonate, nitrate, N-methylglucosamine ammonium salt, 3-hydroxy-2-naphthylcarboxylate, oleate, oxalate, palmitate, bis(hydroxynaphthyl)ate (1,1-methylene-bis-2-hydroxy-3-naphthylcarboxylate, einbonate), pantothenate, phosphate / hydrophosphate, picrates, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, basic acetate, succinate, sulfate, sulfosalicylate, suramate, tannins, tartrates, teoclate, toluenesulfonate, triethyl iodide, and valerate.

[0069] "Patient" or "subject" is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or a non-human primate, such as a monkey, chimpanzee, baboon, or rhesus monkey.

[0070] When used in combination with salt or pharmaceutical compositions, "effective amount" is the amount that is effective in treating or preventing the disease in the subject as described herein.

[0071] As used in this disclosure, the term "carrier" encompasses carriers, excipients, and diluents and refers to materials, compositions, or media that involve carrying or transporting a pharmaceutical agent from one organ or part of a subject's body to another organ or part of the body, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials.

[0072] The term "treatment" in relation to a subject refers to the improvement of at least one symptom of the subject's condition. Treatment includes curing, improving, or at least partially alleviating the condition.

[0073] Unless otherwise indicated, the term “symptom” is used in this disclosure to mean the terms disease, ailment or illness, and is used interchangeably with those terms.

[0074] As used in this disclosure, the terms “administer,” “administering,” or “administration” mean the direct administration of the disclosed salt or composition to a subject, or the administration of a prodrug derivative or analogue of the salt or composition to the subject, wherein the prodrug derivative or analogue can form an equivalent amount of active salt in the subject’s body.

[0075] The salt of this application

[0076] This application relates to salts or their enantiomers, stereoisomers or tautomers that can treat or prevent age-related conditions, and can be used to treat diseases and conditions associated with aging and cell regeneration.

[0077] In some embodiments of the present invention, R a Each time it appears, it is independently H or C1-C6 alkyl. In other embodiments, R a For H. In other implementations, R a It is a C1-C6 alkyl group. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more substituents selected from C1-C6 alkyl groups. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more C2-C6 alkenyl groups. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more C2-C6 ynyl groups. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more (C0-C3 alkylene) C3-C8 cycloalkyl groups. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more (C0-C3 alkylene) heterocyclic alkyl groups. In other embodiments, R a It is formed by one or more (C0-C3 alkylene)C6-C14 Aryl-substituted C1-C6 alkyl groups. In other embodiments, R a It is a C1-C6 alkyl group substituted with one or more (C0-C3 alkylene) heteroaryl groups. In other embodiments, R a It is methyl. In other embodiments, R a It is a methyl group substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl.

[0078] In another implementation, R 1 Independently H, C1-C6 alkyl, C1-C6 haloalkyl, (C0-C3 alkylene)C(O)C1-C6 alkyl, -C(O)OR a -C(O)NR a R b Or -[CH2-CH2-O] k -R a In another implementation, R 1 For H. In another implementation, R 1 It is a C1-C6 alkyl group. In another embodiment, R 1 It is a C1-C6 haloalkyl group. In another embodiment, R 1 It is a (C0-C3 alkylene)C(O)C1-C6 alkyl. In another embodiment, R 1 -C(O)OR a In another implementation, R 1 -[CH2-CH2-O] k -R a In another implementation, R 1 It is a C(O)C1-C6 alkyl group.

[0079] In one implementation, R 2 Independently H, C1-C6 alkyl, C1-C6 haloalkyl, (C0-C3 alkylene)C(O)C1-C6 alkyl, -C(O)OR a -C(O)NR a R b Or -[CH2-CH2-O] k -R a In another implementation, R 2 For H. In another implementation, R 2 It is a C1-C6 alkyl group. In another embodiment, R2 It is a C1-C6 haloalkyl group. In another embodiment, R 2 It is a (C0-C3 alkylene)C(O)C1-C6 alkyl. In another embodiment, R 2 -C(O)OR a In another implementation, R 2 -[CH2-CH2-O] k -R a In another implementation, R 1 It is a C(O)C1-C6 alkyl group.

[0080] In one implementation, R 3 Independently H, C1-C6 alkyl, C1-C6 haloalkyl, (C0-C3 alkylene)C(O)C1-C6 alkyl, -C(O)OR a -C(O)NR a R b Or -[CH2-CH2-O] k -R a In another implementation, R 3 For H. In another implementation, R 3 It is a C1-C6 alkyl group. In another embodiment, R 3 It is a C1-C6 haloalkyl group. In another embodiment, R 3 It is a (C0-C3 alkylene)C(O)C1-C6 alkyl. In another embodiment, R 3 -C(O)OR a In another implementation, R 3 -[CH2-CH2-O] k -R a In another implementation, R 3 It is a C(O)C1-C6 alkyl group.

[0081] In another embodiment of the salt of formula I, R 1 and R 2 Together with the atoms it connects to, it can form a 5-membered heterocycle. In another embodiment of the salt of Formula I, R 1 and R 2 Together with the atoms it connects to, it can form a 6-membered heterocycle. In another embodiment of the salt of Formula I, R 1 and R 2 Together with the atoms attached to it, it can form a 5-membered heterocycle substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C14 Aryl or (C0-C3 alkylene) heteroaryl. In another embodiment of the salt of formula I, R 1 and R 2 Together with the atoms attached to it, it can form a 6-membered heterocycle substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl.

[0082] In another embodiment of the salt of formula I, R 2 and R 3 Together with the atoms it connects to, it can form a 5-membered heterocycle. In another embodiment of the salt of Formula I, R 2 and R 3 Together with the atoms it connects to, it can form a 6-membered heterocycle. In another embodiment of the salt of Formula I, R 2 and R 3 Together with the atoms attached to it, it can form a 5-membered heterocycle substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl. In another embodiment of the salt of formula I, R 2 and R 3 Together with the atoms attached to it, it can form a 6-membered heterocycle substituted with one or more of the following substituents: C1-C6 alkyl, C2-C6 alkenyl, C2-C6 ynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl.

[0083] In another embodiment of the salt of formula I, M 1 It is an amphoteric amino acid. In another embodiment, M 1 For zwitterionic amino acids of formula II:

[0084]

[0085] In one embodiment of the salt of formula I, R 5 H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14Aryl or (C0-C3 alkylene) heteroaryl. In another embodiment, R 5 For H. In another implementation, R 5 It is a C1-C6 alkyl group. In another embodiment, R 5 It is a C2-C6 alkenyl group. In another embodiment, R 5 It is a C2-C6 acetylene group. In another embodiment, R 5 It is a (C0-C3 alkylene)C3–C8 cycloalkyl. In another embodiment, R 5 It is a (C0–C3 alkylene) heterocyclic alkyl group. In another embodiment, R 5 (C0–C3 alkylene)C6–C 14 Aryl. In another embodiment, R 5 It is a (C0–C3 alkylene) heteroaryl. In another embodiment, R 5 It is H, and is formed by one or more (C0–C3 alkylene) SR c Substituted C1–C4 alkyl groups.

[0086] In another implementation, R 5 It is a C1-C6 alkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NRc R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 5 It is a C2-C6 alkenyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 5 It is a C2-C6 alkynyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)ORc (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 5 It is a (C0-C3 alkylene)C3-C8 cycloalkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c)NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 5 It is a (C0-C3 alkylene) heterocyclic alkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0–C3 alkylene)NR c R d (C0–C3 alkylene)OR c (C0–C3 alkylene)OC(O)R c (C0–C3 alkylene)C(O)OR c (C0–C3 alkylene)SR c (C0–C3 alkylene)C(O)SR c (C0–C3 alkylene)SC(O)R c (C0–C3 alkylene)C(O)NR c R d (C0–C3 alkylene)NC(O)NR c R d (C0–C3 alkylene)C(NR) c )NR c R d (C0–C3 alkylene)NR c C(NR c )NR c R d (C0–C3 alkylene)P(O)O n R c R d (C0–C3 alkylene)S(O) m NR c R d (C0–C3 alkylene)S(O) m OR c or (C0–C3 alkylene)BO p R c R d In another implementation, R 5It is a (C0-C3 alkylene) C6-C substituent selected from one or more of the following substituents. 14 Aryl: Cyano, Halogen, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 5 It is a (C0-C3 alkylene) heteroaryl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0–C3 alkylene)NR c R d (C0–C3 alkylene)OR c (C0–C3 alkylene)OC(O)R c (C0–C3 alkylene)C(O)OR c (C0–C3 alkylene)SR c (C0–C3 alkylene)C(O)SR c (C0–C3 alkylene)SC(O)R c (C0–C3 alkylene)C(O)NRc R d (C0–C3 alkylene)NC(O)NR c R d (C0–C3 alkylene)C(NR) c )NR c R d (C0–C3 alkylene)NR c C(NR c )NR c R d (C0–C3 alkylene)P(O)O n R c R d (C0–C3 alkylene)S(O) m NR c R d (C0–C3 alkylene)S(O) m OR c or (C0–C3 alkylene)BO p R c R d .

[0087] In one embodiment of the salt of formula I, R 4 H, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl. In another embodiment, R 4 For H. In another implementation, R 4 It is a C1-C6 alkyl group. In another embodiment, R 4 It is a C2-C6 alkenyl group. In another embodiment, R 4 It is a C2-C6 acetylene group. In another embodiment, R 4 It is a (C0-C3 alkylene)C3–C8 cycloalkyl. In another embodiment, R 4 It is a (C0–C3 alkylene) heterocyclic alkyl group. In another embodiment, R 4 (C0–C3 alkylene)C6–C 14 Aryl. In another embodiment, R 4 It is a (C0–C3 alkylene) heteroaryl group.

[0088] In another implementation, R 4 It is a C1-C6 alkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d(C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 4 It is a C2-C6 alkenyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NRc R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 4 It is a C2-C6 alkynyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c Rd In another implementation, R 4 It is a (C0-C3 alkylene)C3-C8 cycloalkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c R d (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 4 It is a (C0-C3 alkylene) heterocyclic alkyl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0–C3 alkylene)NR c R d (C0–C3 alkylene)OR c (C0–C3 alkylene)OC(O)R c (C0–C3 alkylene)C(O)OR c (C0–C3 alkylene)SR c (C0–C3 alkylene)C(O)SR c (C0–C3 alkylene)SC(O)Rc (C0–C3 alkylene)C(O)NR c R d (C0–C3 alkylene)NC(O)NR c R d (C0–C3 alkylene)C(NR) c )NR c R d (C0–C3 alkylene)NR c C(NR c )NR c R d (C0–C3 alkylene)P(O)O n R c R d (C0–C3 alkylene)S(O) m NR c R d (C0–C3 alkylene)S(O) m OR c or (C0–C3 alkylene)BO p R c R d In another implementation, R 4 It is a (C0-C3 alkylene) C6-C substituent selected from one or more of the following substituents. 14 Aryl: Cyano, Halogen, SeH, (C0-C3 alkylene)NR c R d (C0-C3 alkylene)OR c (C0-C3 alkylene)OC(O)R c (C0-C3 alkylene)C(O)OR c (C0-C3 alkylene)SR c (C0-C3 alkylene)C(O)SR c (C0-C3 alkylene)SC(O)R c (C0-C3 alkylene)C(O)NR c R d (C0-C3 alkylene)NC(O)NR c R d (C0-C3 alkylene)C(NR) c )NR c R d (C0-C3 alkylene)NR c C(NR c )NR c R d (C0-C3 alkylene)P(O)O n R c Rd (C0-C3 alkylene)S(O) m NR c R d (C0-C3 alkylene)S(O) m OR c or (C0-C3 alkylene)BO p R c R d In another implementation, R 4 It is a (C0-C3 alkylene) heteroaryl group substituted with one or more of the following substituents: cyano, halogroup, SeH, (C0–C3 alkylene)NR c R d (C0–C3 alkylene)OR c (C0–C3 alkylene)OC(O)R c (C0–C3 alkylene)C(O)OR c (C0–C3 alkylene)SR c (C0–C3 alkylene)C(O)SR c (C0–C3 alkylene)SC(O)R c (C0–C3 alkylene)C(O)NR c R d (C0–C3 alkylene)NC(O)NR c R d (C0–C3 alkylene)C(NR) c )NR c R d (C0–C3 alkylene)NR c C(NR c )NR c R d (C0–C3 alkylene)P(O)O n R c R d (C0–C3 alkylene)S(O) m NR c R d (C0–C3 alkylene)S(O) m OR c or (C0–C3 alkylene)BO p R c R d .

[0089] In another implementation, R 6 It is H or C1-C6 alkyl. In another embodiment, R 6 For H. In another implementation, R 6It is a C1-C6 alkyl group. In another embodiment, R 6 It is a C1-C6 alkyl group substituted with one or more of the following substituents: cyano, halogroup, (C0-C3 alkylene)NR c R d OR (C0-C3 alkylene) c In another implementation, R 5 For H.

[0090] In yet another implementation scheme, R 5 and R 6 Together with the atoms it connects to, it can form a 5-membered ring. In another embodiment, R 5 and R 6 Together with the atoms attached to it, it can form a 5-membered ring substituted with one or more of the following substituents: cyano, halogroup, (C0-C3 alkylene)NR c R d OR (C0-C3 alkylene) c In another implementation, R 5 and R 6 Together with the atoms it connects to, it can form a 6-membered ring. In another embodiment, R 5 and R 6 Together with the atoms attached to it, it can form a 6-membered ring substituted with one or more of the following substituents: cyano, halogroup, (C0-C3 alkylene)NR c R d OR (C0-C3 alkylene) c .

[0091] In another implementation, R d Each time it appears, it is independently H or C1-C6 alkyl. In another embodiment, R d For H. In another implementation, R d C 1-6 Alkyl group. In another embodiment, R d It is a C1-C6 alkyl group substituted with one or more of the following substituents: (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl.

[0092] In another implementation, R c Each time it appears, it is independently H or C1-C6 alkyl. In another embodiment, R c For H. In another implementation, R c It is a C1-C6 alkyl group. In another embodiment, Rc It is a C1-C6 alkyl group substituted with one or more of the following substituents: (C0-C3 alkylene)C3-C8 cycloalkyl, (C0-C3 alkylene) heterocycloalkyl, (C0-C3 alkylene)C6-C 14 Aryl or (C0-C3 alkylene) heteroaryl.

[0093] In another embodiment, k is 1, 2, 3, 4, 5, 6, 7, or 8 each time it appears. In another embodiment, k is 1. In another embodiment, k is 2. In another embodiment, k is 3. In another embodiment, k is 4. In another embodiment, k is 5. In another embodiment, k is 6. In another embodiment, k is 7. In another embodiment, k is 8.

[0094] In one implementation, m is 0, 1, or 2. In another implementation, m is 0. In another implementation, m is 1. In yet another implementation, m is 2.

[0095] In one implementation, n is 0, 1, or 2. In another implementation, n is 0. In another implementation, n is 1. In yet another implementation, n is 2.

[0096] In one implementation, p is 0, 1, or 2. In another implementation, p is 0. In another implementation, p is 1. In yet another implementation, p is 2.

[0097] In some embodiments of the salt of formula I, the salt has the structure of formula Ia:

[0098]

[0099] In some embodiments of the salt of formula I, the salt has the structure of formula Ib:

[0100]

[0101] In some embodiments of the salt of formula I, the salt has the structure of formula Ic:

[0102]

[0103] In some embodiments of the salt of formula I, the salt has the structure of formula Id:

[0104]

[0105] In some embodiments of the salt of formula I, the salt has the structure of formula Ie:

[0106]

[0107] In some embodiments of the salt of formula I, the salt has the structure of formula If:

[0108]

[0109] In some embodiments of the salt of formula I, the salt has the structure of formula Ig:

[0110]

[0111] In some embodiments of the salt of formula I, the salt has the structure of formula Ih:

[0112]

[0113] In some embodiments of the salt of formula I, the salt has the structure of formula Ii:

[0114]

[0115] In some embodiments of the salt of formula I, the salt has the structure of formula Ij:

[0116]

[0117] In some embodiments of the salt of formula I, the salt has the structure of formula Ik:

[0118]

[0119] In some embodiments of the salt of formula I, the salt has the structure of formula I1:

[0120]

[0121] In some embodiments of the salt of formula I, the salt has the structure of formula Im:

[0122]

[0123] In some embodiments of the salt of formula I, the salt has the structure of formula In:

[0124]

[0125] In some embodiments of the salt of formula I, the salt has the structure of formula Io:

[0126]

[0127] In some embodiments of the salt of formula I, the salt has the structure of formula Ir:

[0128]

[0129] In another implementation, suitable salts include, but are not limited to:

[0130] (S)-2-ammonium-3-phenylpropionate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1) (I-001);

[0131] (2S,3S)-2-ammonium-3-methylpentanoate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onium-3-carboxylate (1:1) (I-002);

[0132] (S)-2-ammonium-3-(1H-indol-3-yl)propionate and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1) (I-003);

[0133] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onthium-3-carboxylate-(S)-6-amino-2-ammonium hexanoate (1:1)(I-004);

[0134] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (S)-5-amino-2-ammonium-5-oxovalerate (1:1) (I-005);

[0135] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onthium-3-carboxylate-(S)-2-ammonium-4-methylpentanoate (1:1)(I-006);

[0136] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onthium-3-carboxylate-(S)-2-ammonium-4-carboxybutyrate (1:1)(I-007);

[0137] (S)-2-Ammonium-3-methylbutyrate-1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onium-3-carboxylate (1:1) (I-008);

[0138] (S)-2-Ammonium-5-guanidinovalerate-1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onyl-3-carboxylate (1:1) (I-009); and

[0139] 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-on-3-carboxylate-(S)-2-ammonium-3-(1H-imidazol-4-yl)propionate (1:1)(I-010).

[0140] Methods for preparing salt

[0141] The salts of this application can be prepared by various methods, including standard chemical methods. Suitable synthetic routes are described in the schemes given below.

[0142] Salts of formula (I) can be prepared by methods known in the field of organic synthesis, which are illustrated in part by the following synthetic schemes. In the schemes described below, it should be fully understood that protecting groups of sensitive or reactive groups are used where necessary, based on general principles or chemistry. Protecting groups are manipulated according to standard methods of organic synthesis (TW Greene and PGM Uts, "Protective Groups in Organic Synthesis", 3rd ed., Wiley, New York 1999). These groups are removed at a convenient stage of salt synthesis using methods readily apparent to those skilled in the art. The selection of procedures, reaction conditions, and the order of their execution should be consistent with the preparation of salts of formula (I).

[0143] Those skilled in the art will recognize the presence of a stereocenter in the salt of formula (I). Therefore, this application includes two possible stereoisomers (unless described in the synthesis), and includes not only racemic salts, but also individual enantiomers and / or diastereomers. When a compound or salt is required as a single enantiomer or diastereomer, it can be obtained by stereospecific synthesis or by resolving the final product or any convenient intermediate. Resolving the final product, intermediate, or starting material can be achieved by any suitable method known in the art. See, for example, "Stereochemistry of Organic Compounds", E.L. Leel, S.H. Wilen, and L.N. Mander (Wiley-Interscience, 1994).

[0144] The salts and compounds described herein can be prepared from commercially available starting materials or synthesized using known organic, inorganic, and / or enzymatic methods.

[0145] The salts of this application can be prepared in a variety of ways well known to those skilled in the art of organic synthesis. For example, the salts of this application can be synthesized using the methods described below, as well as synthetic methods known in the field of synthetic organic chemistry, or variations thereof known to those skilled in the art. These methods include, but are not limited to, those described below. The salts of this application can be synthesized by following the steps outlined in General Scheme 1, which includes different sequences of assembling various intermediates. Starting materials are commercially available or prepared by methods known in reported literature or as shown.

[0146] Option 1

[0147]

[0148] The mixture of enantiomers, diastereomers, and cis / trans isomers produced by the above process can be separated into their individual components by chiral salt techniques, normal-phase, reverse-phase, or chiral column chromatography, depending on the nature of the separation.

[0149] It should be understood that, in the descriptions and formulas shown above, unless otherwise indicated, the group R in the scheme represents R as defined above. 3 Furthermore, for synthetic purposes, the salts of General Scheme 1 are only representative of the selected groups to illustrate the general synthetic method of the salts of Formula (I) as defined herein.

[0150] It should also be understood that the salts disclosed herein have a neutral charge, and the structure of Formula I represents only the genus, which may be balanced with counterions to give the salt a neutral charge if desired. Such counterions may include, but are not limited to, bromide, chloride, and trifluoromethanesulfonate. In one embodiment, the salts of the present invention can be generated in situ without separation from solution. In some embodiments, the salts disclosed herein may be discrete 1:1 or 1:2 salts. In some embodiments, the salts disclosed herein may also be present in other ratios, for example, 1:1.5, 1:5, or 1:10.

[0151] The method of using the disclosed salt

[0152] Another aspect of this disclosure relates to a method for treating or preventing diseases or conditions associated with aging, cellular degeneration, and / or cellular regeneration. Non-limiting examples of such diseases and conditions include infertility, age-related infertility, age-related loss of eye function, decreased bone density, obesity, and insulin insensitivity. In one embodiment, the salt of formula (I) can be used to treat age-related infertility. In another embodiment, the salt of formula (I) can be used to treat fertility.

[0153] Another aspect of this application relates to a method for treating or preventing diseases or conditions associated with aging, cellular degeneration, and / or cellular regeneration. In one embodiment, the salt disclosed herein can be used to treat infertility. In another embodiment...

[0154] The present invention also relates to the use of salts of Formula I and their enantiomers, stereoisomers and tautomers in the manufacture of medicaments for treating aging, cell regeneration, cell degeneration or infertility.

[0155] Another aspect of this disclosure relates to a method for improving the quality and maturation of oocytes or blastocysts. The method includes contacting the oocytes or blastocysts with an IVF medium containing a salt of formula (I) for an effective period of time.

[0156] In another aspect, this disclosure provides a culture medium containing a salt of formula (I). The salt of formula (I) exhibits surprisingly and unexpectedly prolonged stability in solution and is therefore suitable for use in culture media for the period required to enhance NAD+ production before implantation in patients with infertility or age-related infertility. In some embodiments, a culture medium containing a salt of formula (I) is provided. In some embodiments, the culture medium contains various reagents and factors necessary for the oocyte, oocyte, or blastocyst, depending on the stage of maturation and development at which the oocyte, oocyte, or blastocyst is located. For example, the culture medium may contain any of the agents or factors listed in Table 1 below that can be used in IVF culture media:

[0157] Table 1

[0158]

[0159] A cell culture medium for in vitro fertilization is also provided, comprising: one or more salts of formula (I) and a culture medium.

[0160] In one embodiment, the culture medium is an inorganic salt, an energy matrix, amino acids, a chelating agent, a pH indicator, an antibiotic, serum, vitamins, growth factors, or any combination thereof. In one embodiment, the inorganic salt is calcium chloride, magnesium chloride, magnesium sulfate, potassium chloride, sodium bicarbonate, sodium chloride, monosodium phosphate, disodium phosphate, or any combination thereof.

[0161] In one embodiment, the energy matrix is ​​glucose, pyruvate, lactate, pyruvate, or any combination thereof.

[0162] In one embodiment, the amino acid is an essential amino acid. In one embodiment, the essential amino acid is arginine, cysteine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, threonine, tryptophan, tyrosine, valine, or any combination thereof.

[0163] In one implementation, the amino acid is a non-essential amino acid.

[0164] In one embodiment, the non-essential amino acid is alanine, asparagine, aspartic acid, glutamic acid, proline, serine, or any combination thereof.

[0165] In one embodiment, the chelating agent is a cage-like chelate, acetylacetone, aminopolycarboxylic acid, ATMP, BAPTA, BDTH2, citric acid, cryptether, deferasirox, 2,3-dihydrobenzoic acid, 2,3-dimercapto-1-propanesulfonic acid, dimercaptosuccinic acid, DOTA, DTPMP, EDDHA, EDDS, EDTMP, etidronic acid, fura-2, gluconic acid, homocitric acid, iminodiacetic acid, Indo-1, nitrilotriacetic acid, DTPA, phosphonates, phytochelati, polyaspartic acid, sodium polyaspartate, trisodium citrate, transferrin, EDTA, EGTA, or any combination thereof.

[0166] In one embodiment, the pH indicator is phenol red, bromothymol blue, alizarin red, 9-aminoacridine, or any combination thereof.

[0167] In one embodiment, the antibiotic is actinomycin D, ampicillin, carbenicillin, cefotaxime, phosphatidylcholine, gentamicin, kanamycin, neomycin, penicillin, polymyxin B, streptomycin, or any combination thereof.

[0168] In one embodiment, serum is human serum albumin, bovine serum albumin, fetal bovine serum, synthetic serum, or any combination thereof.

[0169] In one embodiment, the vitamins are ascorbic acid, biotin, sodium menadione bisulfite, mitomycin C, pyridoxine dihydrochloride, retinyl acetate, (-)-riboflavin, (+)-L-ascorbic acid sodium, (+)-α-tocopherol, and vitamin B. 12 Thiamine hydrochloride, inositol, pyridoxal hydrochloride, nicotinamide, folic acid, D-calcium pantothenate, choline chloride, or any combination thereof.

[0170] In one embodiment, the growth factor is adrenaline medullaris, angiopoietin, bone morphogenetic protein, macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), epidermal growth factor, hepatocyte ligand (ephrin), erythropoietin, fibroblast growth factor, growth differentiation factor-9, hepatocyte growth factor, insulin, insulin-like growth factor, interleukin, keratinocyte growth factor, migration stimulating factor, macrophage stimulating protein, myostatin, neurotrophic factor, T-cell growth factor, thrombopoietin, transforming growth factor, tumor necrosis factor-α, vascular endothelial growth factor, or any combination thereof.

[0171] In one embodiment, the cell culture medium further comprises oocytes, fertilized eggs, blastocysts, or any combination thereof.

[0172] In addition, kits for IVF culture media are provided, which contain various agents and factors necessary for oocyte or blastocyst maturation, including one or more salts of formula (I). These agents and cofactors are soluble in solution for use in producing IVF culture media shortly before exposing oocytes or blastocysts to implantation in patients requiring treatment for infertility or age-related infertility.

[0173] The present invention also relates to the use of salts of Formula I and their enantiomers, stereoisomers, and tautomers in the manufacture of medicaments for treating aging, cell regeneration, cell degeneration, or infertility. In some embodiments, the infertility being treated is age-related infertility.

[0174] Another aspect of the invention is a pharmaceutical composition comprising a salt of formula I and a pharmaceutically acceptable carrier.

[0175] Another aspect of the invention is a pharmaceutical composition comprising a salt of formula I and a pharmaceutically acceptable carrier comprising a therapeutically effective amount of one or more other therapeutic agents.

[0176] In some embodiments, administration of a salt of formula (I) or a pharmaceutical composition comprising the salt of the present invention and a pharmaceutically acceptable carrier induces changes in cell cycle or cell viability.

[0177] In some embodiments, administration of a salt of formula (I) or a pharmaceutical composition comprising the salt of the present invention and a pharmaceutically acceptable carrier induces preventative changes in age-related symptoms or diseases.

[0178] The salt of the present invention disclosed herein can be applied in an effective amount to treat or prevent symptoms in a subject and / or prevent the development of age-related symptoms or diseases in the subject.

[0179] The disclosed salt can be administered via any mode of therapeutic application. These modes include systemic or local administration, such as oral, nasal, parenteral, transdermal, subcutaneous, vaginal, buccal, rectal, or topical administration.

[0180] Depending on the intended mode of administration, the disclosed compositions may be in solid, semi-solid, or liquid dosage forms, such as injections, tablets, suppositories, pills, time-release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, etc., sometimes in unit doses and consistent with conventional pharmaceutical practices. Similarly, they may be administered intravenously (push and infusion), intraperitoneally, subcutaneously, or intramuscularly, all in forms well known to those skilled in the art of pharmacy.

[0181] When used for the indicated effects, the effective dose range of the disclosed salt is from about 0.5 mg to about 5000 mg for the therapeutic effect required by the disclosed salt. Compositions for in vivo or in vitro use may contain about 0.5, 5, 20, 50, 75, 100, 150, 250, 500, 750, 1000, 1250, 2500, 3500, or 5000 mg, or amounts from one to another in the dosage list, of the disclosed salt. In one embodiment, the composition is in the form of a scoreable tablet.

[0182] Dosing regimens using the disclosed salts are selected based on a number of factors, including patient type, species, age, weight, sex, and medical condition; the severity of the condition to be treated; route of administration; patient's renal or hepatic function; and the specific disclosed salt used. Physicians or veterinarians with ordinary skill in the art can readily determine and prescribe the effective amount of the drug required to prevent, resist, or halt the progression of the condition.

[0183] The illustrative pharmaceutical compositions are tablets and gelatin capsules comprising the salts of the present invention and pharmaceutically acceptable carriers, such as a) diluents, such as purified water, triglyceride oils such as hydrogenated or partially hydrogenated vegetable oils or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oil (such as EPA or DHA) or esters or triglycerides thereof or mixtures thereof, omega-3 fatty acids or derivatives thereof, lactose, dextran, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin, glucose and / or glycine; b) lubricants, such as silica, talc, stearic acid, its magnesium or calcium salts, sodium oleate, sodium stearate, and so on. Magnesium fatty acid, sodium benzoate, sodium acetate, sodium chloride and / or polyethylene glycol; also used in tablets; c) binders, such as magnesium aluminum silicate, starch paste, gelatin, astragalus gum, methylcellulose, sodium carboxymethyl cellulose, magnesium carbonate, natural sugars (such as glucose or β-lactose), corn sweeteners, natural and synthetic gums such as gum arabic, astragalus gum or sodium alginate, waxes and / or polyvinylpyrrolidone, if needed; d) disintegrants, such as starch, agar, methylcellulose, bentonite, xanthan gum, alginate or its sodium salt, or effervescent mixtures; e) absorbents, colorants, flavorings and sweeteners; f) emulsifiers or dispersants, such as Tween 80. Labrasol, HPMC, DOSS, Caproyl 909, Labrafac, Labrafil, Peceol, Transcutol, Capmul MCM, Capmul PG-12, Captex 355, Gelucire, Vitamin E TGPS or other acceptable emulsifiers; and / or g) agents that enhance salt absorption, such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400, PEG200.

[0184] Most amino acids possess buffering properties similar to salts, and their multiple pKa values ​​result in a wider range of stable pH values ​​that are more compatible with biological fluids, making them more suitable for intravenous administration. Naturally occurring amino acids, NMN, and NaMN are endogenous substances. Therefore, such mixtures are unlikely to be toxic to mammals. Some products may have enhanced solubility and solid-form stability. In one embodiment, the salt may have enhanced water solubility. In one embodiment, the salt may have enhanced solid-form stability. In one embodiment, the salt may have enhanced chemical stability.

[0185] Example

[0186] This disclosure is further illustrated by the following embodiments and synthetic schemes, which should not be construed as limiting the scope or spirit of this disclosure to the specific procedures described herein. It should be understood that the embodiments are provided to illustrate certain implementations and are not intended to limit the scope of this disclosure. It should be further understood that various other implementations, modifications, and equivalents may be conceived by those skilled in the art without departing from the spirit of this disclosure and / or the scope of the appended claims.

[0187] The following salts disclosed herein are prepared using general synthetic methods, including but not limited to reagents such as valine, leucine, alanine, isoleucine, methionine, phenylalanine, tryptophan, and tyrosine. Suitable solvents such as methanol, ethanol, water, acetic acid, ethylene glycol, and isopropanol are also used.

[0188] The abbreviations used in the following embodiments and elsewhere herein are:

[0189] AcOH (acetic acid)

[0190] anh. (anh) without water

[0191] atm atmosphere

[0192] aq. water-based

[0193] br wide peak

[0194] Boc tert-butoxycarbonyl

[0195] Salt water saturated aqueous sodium chloride

[0196] n-BuLi n-Butyllithium

[0197] n-BuOH n-Butanol

[0198] Calc'd (calculated value)

[0199] CDCl3 (deuterated chloroform)

[0200] CDI carbonyl diimidazole

[0201] Chloroform-d deuterated chloroform

[0202] d Double peak

[0203] dd Two double peaks

[0204] dt Two triplet peaks

[0205] D2O (deuterated water, deuterium oxide)

[0206] DCE dichloroethane

[0207] DCM dichloromethane

[0208] DIAD (Diisopropyl Azodicarbonate)

[0209] DIPEA N,N-Diisopropylethylamine

[0210] DMAc N,N-dimethylacetamide

[0211] DMAP N,N-dimethylpyridine-4-amine

[0212] DME 1,2-dimethoxyethane

[0213] DMEDA N,N′-Dimethylethylenediamine

[0214] DMF N,N-dimethylformamide

[0215] DMSO (dimethyl sulfoxide)

[0216] DMSO-d6 Deuterated dimethyl sulfoxide

[0217] EDA (ethylenediamine)

[0218] EDC 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide

[0219] Et2O diethyl ether

[0220] EtOAc (ethyl acetate)

[0221] EtOH (ethanol)

[0222] ESI Electrospray Ionization

[0223] g gram

[0224] h hours

[0225] H hydrogen

[0226] 1 H NMR (Nuclear Magnetic Resonance) (Proton Nucleus)

[0227] HATU [bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0228] HBTU 3-[bis(dimethylamino)methylene]-3H-benzotriazole-1-oxide hexafluorophosphate

[0229] HOBt Hydroxybenzotriazole

[0230] HPLC (High-Performance Liquid Chromatography)

[0231] Hz Hertz

[0232] J coupling constant

[0233] KHCO3 (potassium bicarbonate)

[0234] KHMDS (Potassium Hexamethyldisilazane)

[0235] KOAc potassium acetate

[0236] LCMS (Liquid Chromatography-Mass Spectrometry)

[0237] LHMDS (Lithium Hexamethyldisilazane)

[0238] [#]M molar concentration

[0239] m multiplet

[0240] [M+H] + Molecular ion hydrogenation

[0241] [M-tBu+H] + Molecular ion tert-butyl hydrogenation

[0242] mCPBA (m-chloroperoxybenzoic acid)

[0243] Me2NH dimethylamine

[0244] Me4NBr Tetramethylammonium bromide

[0245] MeCN Acetonitrile

[0246] MeNH2 methylamine

[0247] MeOH (methanol)

[0248] Methanol-d4 deuterated methanol

[0249] 2-MeTHF 2-Methyltetrahydrofuran

[0250] mg

[0251] MHz

[0252] min minutes

[0253] mmol millimole

[0254] mL

[0255] MS mass spectrometry

[0256] MS ES mass spectrometry electrospray ionization

[0257] Ms2O methanesulfonic anhydride

[0258] MTBE (methyl tert-butyl ether)

[0259] MW microwave

[0260] m / z mass-to-charge ratio

[0261] μL

[0262] N2 nitrogen

[0263] NaHCO3 (Sodium bicarbonate)

[0264] NaMN (Nicotinic Acid Mononucleotide)

[0265] NIS N-iodosuccinimide

[0266] NMP (N-methyl-2-pyrrolidone)

[0267] NMR (Nuclear Magnetic Resonance)

[0268] PEPPSI-iPr [1,3-bis(2,6-diisopropylphenyl)imidazol-2-ylidene](3-chloropyridinyl)palladium(II) dichloride

[0269] PdCl2(Amphos) bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II)

[0270] Pd2(dba)3 tris(dibenzylacetone)dipalladium(0)

[0271] Pd(OAc)₂ Palladium(II) acetate

[0272] PdCl2(dppf) [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloro

[0273] PdCl2(MeCN)2 bis(acetonitrile)dichloropalladium(II)

[0274] PdCl2(PPh3)2 bis(triphenylphosphine palladium(II) dichloride

[0275] Pd(P(Cy)3)2Cl2 dichlorobis(tricyclohexylphosphine)palladium(II)

[0276] Pd(PPh3)4 tetrakis(triphenylphosphine)palladium(0)

[0277] Pd(t-Bu3P)2 bis(tri-tert-butylphosphine)palladium(0)

[0278] pH

[0279] PMB 4-Methoxybenzyl

[0280] PMBCl 4-Methoxybenzyl chloride

[0281] ppm (parts per million)

[0282] prep preparation

[0283] pyridine

[0284] q Quadruple Peak

[0285] qd Four double peaks

[0286] quant.

[0287] Quin. Five-peak

[0288] Quind five double peaks

[0289] RBF round-bottom flask

[0290] Rt retention time

[0291] rt room temperature

[0292] s Single peak

[0293] sat. saturated

[0294] sat.aq. saturated aqueous solution

[0295] SEMCl 2-(trimethylsilyl)ethoxymethyl chloride

[0296] t triple peak

[0297] t-BuLi tert-butyllithium

[0298] td Three double peaks

[0299] TMS (trimethylsilyl)

[0300] TMSCl trimethylsilyl chloride

[0301] tt Three triple peaks

[0302] T3P polyphosphonic anhydride

[0303] TBAB Tetrabutylammonium Bromide

[0304] TEA Triethylamine

[0305] TFA (trifluoroacetic acid)

[0306] TFAA (trifluoroacetic anhydride)

[0307] THF Tetrahydrofuran

[0308] TLC (Thin Layer Chromatography)

[0309] TPPO (triphenylphosphine oxide)

[0310] XantPhos 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthracene

[0311] XPhos 2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

[0312] Example 1. Synthesis of (S)-2-ammonium-3-phenylpropionate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1)

[0313]

[0314] Add NaR (0.100 g, 0.392 mmol, 1 equivalent) and 10 mL of distilled deionized water to 50 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-phenylalanine (0.0647 g, 392 mmol, 1.0 equivalent) to this solution in one step. After this addition, the pH is ~4.9-5.2, and all solids have dissolved. Then remove the flask and freeze the colorless solution using liquid nitrogen. Once the flask is frozen, connect it to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a colorless solid.

[0315] Yield: 160.6 mg (98%)

[0316] Melting point: 130℃-133℃ (corrected, degradation), degassed at 145℃

[0317] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.47(s,1H),9.18(d,1H),8.96(d,1H),8.21(dd,1H ),7.48-7.32(m,5H),6.25(d,1H),4.51(m,2H),4.38(t,1H),4.10-4.00(2x dd,2H),3.92(dd,1H),3.31(dd,1H),3.15(dd,1H)ppm

[0318] Example 2. Synthesis of (2S,3S)-2-ammonium-3-methylpentanoate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1)

[0319]

[0320] Add NaR (0.100 g, 0.392 mmol, 1 equivalent) and 20 mL of distilled deionized water to 50 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-isoleucine (0.080 g, 392 mmol, 1.0 equivalent) to this solution in a single addition. After this addition, the pH is ~4.6–4.9, and all solids have dissolved.

[0321] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a colorless solid.

[0322] Yield: 147.1 mg (97%)

[0323] Melting point: 144℃-148℃ (corrected for degradation)

[0324] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.48(s,1H),9.18(d,1H),8.98(d,1H),8.21(dd,1H),6.24(d,1H),4.48-4.55(m,2H),4.38(t,1H), 4.07(dd,1H),3.93(dd,1H),3.69(d,1H),2.00(m,1H),1.55-1.45(m,1H),1.35-1.24(m,1H),1.04(d,3H),0.95(t,3H)ppm

[0325] Example 3. Synthesis of (S)-2-ammonium-3-(1H-indol-3-yl)propionate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1)

[0326]

[0327] Add NaR (0.100 g, 0.392 mmol, 1 equivalent) and 30 mL of distilled deionized water to 50 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-tryptophan (0.080 g, 392 mmol, 1.0 equivalent) to this solution in a single addition. After this addition, the pH is ~4.9–5.2, and all solids have dissolved.

[0328] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a yellow solid.

[0329] Yield: 171.8 mg (96%)

[0330] Melting point: 117℃-126℃ (corrected, degradation), degassed at 133℃

[0331] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.42(s,1H),9.12(d,1H),8.93(d,1H),8.15(dd,1H),7.72(d,1H),7.53(d,1H),7.30(s+t,2H), 7.20(t,1H),6.28(d,1H),4.481(m,2H),4.38(t,1H),4.10-4.00(m,2H),3.92(dd,1H),3.51(dd,1H),3.32(dd,1H)ppm

[0332] Example 4. Synthesis of 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onthium-3-carboxylate-(S)-6-amino-2-ammonium hexanoate (1:1)

[0333]

[0334] Add NaR (0.200 g, 0.784 mmol, 1 equivalent) and 40 mL of distilled deionized water to 250 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-lysine (0.115 g, 0.744 mmol, 0.95 equivalent) to this solution in a single addition. After this addition, the pH is 9.63, and all solids have dissolved.

[0335] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a yellow to brown solid.

[0336] Production: Quantitative

[0337] Analyze the data. 1H-NMR(400MHz,D2O)δ=9.43(s,1H),9.13(d,1H),8.92(d,1H),8.17(dd,1H),6.21(d,1H),4.48(m,2H), 4.33(t,1H),4.02(dd,1H),3.88(dd,1H),3.46(t,1H),2.98(d,2H),1.8-1.6(m,4H),1.5-1.3(m,2H)ppm

[0338] Example 5.1 Synthesis of ((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onthium-3-carboxylate (S)-5-amino-2-ammonium-5-oxopentanoate (1:1)

[0339]

[0340] Add NaR (0.200 g, 0.784 mmol, 1 equivalent) and 15 mL of distilled deionized water to 100 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-glutamine (0.109 g, 0.744 mmol, 0.95 equivalent) to this solution in a single addition. After this addition, the pH is 4.95, and all solids have dissolved.

[0341] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a yellow to brown solid.

[0342] Production: Quantitative

[0343] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.45(s,1H),9.13(d,1H),8.92(d,1H),8.17(dd,1H),6.20(d,1H),4.50-4.43(m ,2H),4.33(t,1H),4.03(dd,1H),3.89(dd,1H),3.75(t,1H),2.50-2.37(ddd,2H),2.15-2.07(m,2H)ppm

[0344] Example 6.1 - ((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onthium-3-carboxylate-(S)-2-ammonium-4-methylpentanoate (1:1)

[0345]

[0346] Add NaR (0.200 g, 0.784 mmol, 1 equivalent) and 15 mL of distilled deionized water to 100 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-leucine (0.098 g, 0.744 mmol, 0.95 equivalent) to this solution in a single addition. After this addition, the pH is 5.61, and all solids have dissolved.

[0347] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product is colorless.

[0348] Production: Quantitative

[0349] Analyze the data. 1 H-NMR (400MHz, D2O) δ = 9.43 (s, 1H), 9.12 (d, 1H), 8.92 (d, 1H), 8.17 (dd, 1H), 6.2 1(d,1H),4.50-4.43(m,2H),4.34(t,1H),4.03(dd,1H),3.89(dd,1H),3.71(app t,1H),1.8-1.6(m,3H),0.98-0.9(2x d,6H)ppm

[0350] Example 7.1 Synthesis of ((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onthium-3-carboxylate-(S)-2-ammonium-4-carboxybutyrate (1:1)

[0351]

[0352] Add NaR (0.200 g, 0.784 mmol, 1 equivalent) and 15 mL of distilled deionized water to 100 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-glutamic acid (0.110 g, 0.744 mmol, 0.95 equivalent) to this solution in a single addition. After this addition, the pH is 3.52, and all solids have dissolved.

[0353] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product is colorless.

[0354] Production: Quantitative

[0355] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.45(app m,1H),9.15(app m,1H),8.94(appm,1H),8.18(app m,1H),6.21(app m,1H),4.50-4.43(m,2H),4.34(m,1H),4.05(m,1H),3.89(m,1H),3.78(app m,1H),2.55(m,2H),2.25-2.15(m,2H)ppm

[0356] Example 8. Synthesis of (S)-2-ammonium-3-methylbutyrate-1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1)

[0357]

[0358] Add NaR (0.200 g, 0.784 mmol, 1 equivalent) and 15 mL of distilled deionized water to 100 mL of 1N RBF equipped with a water condenser, and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-valine (0.087 g, 0.744 mmol, 0.95 equivalent) to this solution in a single addition. After this addition, the pH is 5.67, and all solids have dissolved.

[0359] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product is colorless.

[0360] Production: Quantitative

[0361] Analyze the data. 1 H-NMR (400MHz, D2O) δ = 9.43 (s, 1H), 9.13 (d, 1H), 8.92 (dt, 1H), 8.17 (dd, 1H), 6.21 (d, 1H), 4.50-4. 43(m,2H),4.34(t,1H),4.03(dd,1H),3.89(dd,1H),3.59(d,1H),2.30-2.20(m,1H),1.03,0.97(2x d,6H) ppm

[0362] Example 9. Synthesis of (S)-2-ammonium-5-guanidinovalerate-1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1)

[0363]

[0364] Add NaR (0.25 g, 0.979 mmol, 1 equivalent) and 25 mL of distilled deionized water to 100 mL of 3N RBF and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-arginine (0.152 g, 0.979 mmol, 1.0 equivalent) to this solution in a single addition.

[0365] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a yellow solid.

[0366] Production: Quantitative

[0367] Analyze the data. 1 H-NMR(400MHz,D2O)δ=9.42(s,1H),9.12(d,1H),8.93(d,1H),8.15(dd,1H),6.28(d,1H),4 .481(m,2H),4.38(t,1H),4.10-3.9(dq,2H),3.3(m,1H),3.18(dd,1H).1.5-1.8(m,4H)ppm

[0368] Example 10.1 Synthesis of ((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onthium-3-carboxylate-(S)-2-ammonium-3-(1H-imidazol-4-yl)propionate (1:1)

[0369]

[0370] Add NaR (0.25 g, 0.979 mmol, 1 equivalent) and 25 mL of distilled deionized water to 100 mL of 3N RBF and mix to form a solution, or a weak suspension. Cool this solution using an ice / water bath. Then add L-histidine (0.152 g, 0.979 mmol, 1.0 equivalent) to this solution in a single batch.

[0371] The flask was then removed and the colorless solution was frozen using liquid nitrogen. Once frozen, the flask was connected to a freeze dryer. This will slowly remove the water. Once dried, the product appears as a colorless solid.

[0372] Production: Quantitative.

[0373] Analyze the data. 1H-NMR(400MHz,D2O)δ=9.42(s,1H),9.12(d,1H),8.93(d,1H),8.15(dd,1H),7.72(d,1H),7.05( s,1H),6.28(d,1H),4.481(m,2H),4.38(t,1H),4.10-3.9(dt,2H),4.0(q,4H),3.18(dd,2H)ppm.

[0374] Example 11. NAD cell assay

[0375] NAD levels were determined using the NAD cycling method described by Zhu and Rand, PLoS One (2012), which is incorporated herein by reference. COV434 cells were maintained in 6-well plates and treated with the indicated compound at a concentration of 200 μM for 4 h. The culture medium was removed, the plates were washed in cold PBS, and the cells were scraped from the NAD extraction buffer containing 10 mM nicotinamide, 50 mM Tris HCl, and 0.1% Triton X-100. The cells were homogenized by sonication for 5 sec, and the sample was centrifuged at 7,000 g for 5 min at 4°C. Aliquots were taken for subsequent protein assays, and the samples were then passed through a 10 kDa amicon filter at 14,000 g for 30 min at 4°C to remove proteins from the samples. Each sample was measured in triplicate, with 25 μL of sample added to 100 μL of ADH cycling mixture (0.2 mg / mL alcohol dehydrogenase, 2% ethanol, 100 mM Tris, pH 8.5). The sample was circulated at room temperature for 10 minutes, followed by the addition of 50 μL of MTT / PMS solution (0.1 mM methyl phenazine sulfate, 0.8 mM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), 100 mM Tris-HCl, pH 8.5). The plate was then incubated for 15 minutes, and absorbance was measured at 570 nM. NAD concentration was deduced from the standard curve and normalized to the protein concentration determined by BCA protein assay.

[0376] The results of the above measurements are shown in Table 2 (below). Adjusted fold increases were obtained by direct comparison of the salts compared to their parent counterparts in molar amounts. In other words, based on the same amount (molar) of salt tested in cells, compounds of this disclosure derived from the NaR parent molecule will only have their NAD activity levels measured against NaR. For example, the fold increase of compound I-002 is based on NAD activity observed in direct comparison with the same molar amount in cells as the NaR parent. Similarly, the fold increase of I-003 is based on direct comparison with NaR.

[0377] Table 2

[0378]

[0379] Equivalent solution

[0380] Using only conventional experiments, those skilled in the art will recognize or be able to identify numerous equivalents of the particular embodiments specifically described herein. These equivalents are intended to be covered within the scope of the following claims.

Claims

1. A salt selected from the group consisting of: (2S,3S)-2-ammonium-3-methylpentanoate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridin-1-onium-3-carboxylate (1:1) (I-002); and (S)-2-ammonium-3-(1H-indol-3-yl)propionate compound and 1-((2R,3R,4S,5R)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)pyridine-1-onium-3-carboxylate (1:1) (I-003).

2. The salt according to claim 1, having the following structure:

3. The salt according to claim 1, having the following structure:

4. A pharmaceutical composition comprising a salt as described in any one of claims 1 to 3 and a pharmaceutically acceptable carrier.

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