Crystalline solids of nicotinic acid mononucleotide and its esters and methods of making and using them

Abstract

The present disclosure relates to crystalline solids comprising a compound of formula (I) where R is n-propyl, and methods of making compounds of formula (I) where R is C1-C4 alkyl or C2-C4 alkenyl. The present disclosure also relates to crystalline solids comprising a compound of formula (II). The present disclosure further relates to methods of preparing the crystalline solids, and pharmaceutical preparations of the crystalline solids, and the use of such pharmaceutical preparations in the treatment of diseases and conditions. [Formula 1] TIFF2024520410000022.tif94129

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Application No. 63 / 193,905, filed May 27, 2021, which is incorporated by reference in its entirety for all purposes. [Background technology]

[0002] Nicotinamide adenine dinucleotide (NAD) and related compounds are known as essential coenzymes in cellular redox reactions in all living organisms. Several lines of evidence also indicate that NAD is involved in several important signaling pathways in mammalian cells, including poly(ADP-ribosyl)ation in DNA repair, mono-ADP-ribosylation in immune response and G protein-coupled signaling, as well as the synthesis of cyclic ADP-ribose and nicotinic acid adenine dinucleotide phosphate (NAADP) in intracellular calcium signaling. It has also been shown that NAD and its metabolites play an important role in transcriptional regulation. In particular, the discovery of Sir2 NAD-dependent deacetylase activity has drawn attention to this role of NAD. Despite advances in the biological understanding of NAD, there remains a need for improved compositions and methods of using such compositions for pharmacological intervention and / or manipulation of the NAD pathway in living cells and tissues.

[0003] Nicotinic acid mononucleotide (also known as nicotinic acid ribonucleotide) and certain nicotinic acid mononucleotide derivatives are believed to increase cellular NAD production (Sauve, U.S. Pat. No. 10,961,268(B2)). However, these compounds are difficult to synthesize on a pharma- ceutically relevant scale with sufficient purity. Given the therapeutic benefits associated with nicotinic acid mononucleotide and its derivatives, there is a need for improved compositions and methods for preparing such compositions. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] U.S. Patent No. 10,961,268(B2) Summary of the Invention

[0005] The present disclosure relates to compounds, crystalline solids, and compositions of compounds and / or crystalline solids for modulating nicotinamide adenine dinucleotide (NAD, also referred to as NAD+ in its oxidized form and NADH in its reduced form).

[0006] One aspect of the present disclosure relates to a crystalline solid comprising a compound of formula (I): [ka] In the formula, R is n-propyl (compound 1).

[0007] A further aspect of the present disclosure relates to a crystalline solid comprising a compound of formula (II). [ka]

[0008] In some embodiments, the disclosure relates to such compounds, crystalline solids, and compositions, as well as methods of making compounds and compositions of formula (I) where R is C1-C4 alkyl or C2-C4 alkenyl. In some embodiments, the disclosure relates to pharmaceutical compositions containing one or more NAD modulating compounds and / or crystalline solids as a first component in combination with one or more active pharmaceutical ingredients. In further embodiments, the disclosure relates to methods of using such compounds, crystalline solids, and / or compositions to promote increased intracellular levels of nicotinamide adenine dinucleotide (NAD) in cells and tissues to treat disease and / or improve cell and tissue viability. [Brief description of the drawings]

[0009] [Figure 1A]1 shows an experimentally obtained XRD pattern of compound 1 as a crystalline solid. [Figure 1B] The top pattern shows an overlay of the experimental diffractogram of compound 1 at room temperature and the bottom pattern shows the calculated diffractogram of compound 1 simulated at 100 K. The slight differences between the simulated and experimental diffractograms are due to lattice changes due to temperature and preferred orientation. [Figure 1C] FIG. 1 is a diagram of the crystal lattice unit cell of Compound 1. [Figure 2A] 1 shows an experimentally obtained XRD pattern of compound 2 as a crystalline solid. [Figure 2B] 1 shows a simulated XRD pattern of compound 2 as a crystalline solid. [Figure 2C] The top pattern shows an overlay of the experimental diffractogram of compound 2, and the bottom pattern shows the calculated diffractogram from the single crystal X-ray structure. The slight differences between the simulated and experimental diffractograms are due to lattice changes due to temperature and preferred orientation. [Diagram 3] FIG. 1 shows an experimentally obtained proton NMR spectrum of compound 1 in DMSO-d6. The X-axis shows the chemical shift (ppm). [Figure 4] FIG. 1 shows an experimentally obtained proton NMR spectrum of compound 2 in D2O. The X-axis shows the chemical shift (ppm). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] definition Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by one of ordinary skill in the art to which this disclosure pertains. As used herein, the following terms have the meanings ascribed to them below, unless otherwise specified.

[0011] In this disclosure, "comprises," "comprising," "containing," "having," and the like can have the meaning ascribed to them in U.S. Patent Law, and can mean "includes," "including," and similarly, "consisting essentially of" or "consists essentially" and the like can have the meaning ascribed to them in U.S. Patent Law, and the terms are open-ended, permitting the presence of more than what is recited so long as the basic or novel characteristics of what is recited are not altered by the presence of more than what is recited, but excluding prior art embodiments.

[0012] Ranges given herein are understood to be shorthand for all values ​​within that range, for example, a range of 1 to 50 is understood to include any number, combination of numbers, or subranges from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

[0013] As used herein, the phrase "a" or "an" entity refers to one or more of that entity, e.g., a compound refers to one or more compounds or at least one compound. Thus, the terms "a" (or "an"), "one or more," and "at least one" may be used interchangeably herein.

[0014] As used herein, unless otherwise clearly stated or apparent from the context, the term "about" is understood to be within normal tolerances in the art, e.g., within 2 standard deviations of the mean. About may be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from the context, all numerical values ​​provided herein are modified by the term about.

[0015] The term "alkyl" as used herein refers to a branched or unbranched saturated hydrocarbon group of 1 to about 20 carbon atoms, preferably 1 to about 10 carbon atoms. Examples of straight-chain and branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. Alkyl groups can be cyclic or non-cyclic. Alkyl groups can be branched or unbranched (i.e., straight-chain). A "lower alkyl" group is an alkyl group containing 1 to 6 (e.g., 1 to 4) carbon atoms.

[0016] The term "alkenyl," as used herein, refers to an aliphatic group containing at least one double bond.

[0017] As used herein, the term "optional" or "optionally" means that the subsequently described event or circumstance may occur, but need not occur, and that the description includes instances in which the event or circumstance occurs and instances in which it does not occur. For example, "optional bond" means that the bond may or may not be present, and that the description includes single bonds, double bonds, or triple bonds.

[0018] The term "purified" refers to the purity of a given compound, as described herein. For example, a given compound is "purified" when it is the major component of a composition, i.e., when it is at least about 50% w / w pure. Thus, "purified" includes at least about 50% w / w purity, at least about 60% w / w purity, at least about 70% purity, at least about 80% purity, at least about 85% purity, at least about 90% purity, at least about 92% purity, at least about 94% purity, at least about 96% purity, at least about 97% purity, at least about 98% purity, at least about 99% purity, at least about 99.5% purity, and at least about 99.9% purity, and "substantially pure" includes at least about 97% purity, at least about 98% purity, at least about 99% purity, at least about 99.5% purity, and at least about 99.9% purity.

[0019] The term "metabolite," as described herein, refers to a compound produced in vivo following administration to a subject.

[0020] The term "salt," as described herein, refers to a compound comprising a cation and an anion, which may be produced by protonation of a proton accepting moiety and / or deprotonation of a proton donating moiety. Note that protonation of a proton accepting moiety results in the formation of a cationic species whose charge is balanced by the presence of physiological anions, and deprotonation of a proton donating moiety results in the formation of an anionic species whose charge is balanced by the presence of physiological cations.

[0021] The phrase "pharmaceutically acceptable salt" means a salt that is pharma- ceutically acceptable. Examples of pharma-ceutically acceptable salts include (1) acid addition salts formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with organic acids, such as acetic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, laurylsulfuric acid, gluconic acid, glutamic acid, salicylic acid, muconic acid, and the like, or (2) base addition salts formed with a conjugate base of any of the above inorganic acids, where the conjugate base is Na + , K + , Mg 2+ , Ca 2+ , N.H. g R 4-g + and R is selected from the group consisting of C 1-3 and g is a number selected from 0, 1, 2, 3, or 4. It should be understood that all references to pharma-ceutically acceptable salts include the solvent addition forms (solvates) of the same acid addition salt or crystalline solids as defined herein.

[0022] The present disclosure also includes useful forms of the compounds of the present disclosure, such as metabolites, solvates, prodrugs, salts, particularly pharma- ceutically acceptable salts, and / or co-precipitates.

[0023] The compounds of the present disclosure can exist as solvates, for example, the compounds of the present disclosure form crystals that contain molecules of polar solvents, such as water, methanol or ethanol, as structural elements of the crystal lattice of the compound. The molecules of polar solvents may be present in stoichiometric or non-stoichiometric ratios with the molecules of the compound. In the case of stoichiometric solvates, for example, hemisolvates, (semi)solvates, monosolvates, sesquisolvates, disolvates, trisolvates, tetrasolvates, pentasolvates, etc. are possible. The present disclosure includes all such solvates.

[0024] Furthermore, the compounds of the present disclosure can exist in free form, e.g., as a free base, or as a free acid, or as a zwitterion, or in the form of a salt, which can be any salt, either an organic or inorganic addition salt, in particular any pharma- ceutically acceptable organic or inorganic addition salt customarily used in pharmacology or used, for example, to isolate or purify the compounds of the present disclosure.

[0025] The term "subject" to which administration is contemplated includes, but is not limited to, humans (i.e., male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or an adult subject (e.g., young adult, middle-aged adult, or elderly adult)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys), mammals, including commercially relevant mammals such as cows, pigs, horses, sheep, goats, cats, and / or dogs, and / or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and / or turkeys.

[0026] The terms "treatment," "treating," "alleviating," and "ameliorating" are used interchangeably herein. These terms refer to an approach to obtain a beneficial or desired result, including but not limited to therapeutic benefit and / or prophylactic benefit. Therapeutic benefit refers to the eradication or amelioration of the underlying disorder being treated. Therapeutic benefit is also achieved by eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder, such that an improvement is observed in the patient, even though the patient may still be affected by the underlying disorder. For prophylactic benefit, pharmaceutical compounds and / or compositions may be administered to patients at risk of developing a particular disease, or to patients who report one or more of the physiological symptoms of the disease, even if the disease has not been diagnosed.

[0027] As used herein, a therapeutic agent that "prevents" a disorder or condition refers to a compound and / or a crystalline solid thereof that, in a statistical sample, reduces the onset of the disorder or condition in a treated sample compared to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition compared to an untreated control sample.

[0028] The term "treating" includes prophylactic and / or therapeutic treatment. The term "prophylactic or therapeutic" treatment is art-recognized and includes administration of one or more of the disclosed compositions to a subject. When administered prior to clinical manifestation of an undesirable condition (e.g., a disease or other undesirable condition in a subject), the treatment is prophylactic (i.e., it protects the subject from the onset of the undesirable condition), whereas when administered after the manifestation of an undesirable condition, the treatment is therapeutic (i.e., it is intended to reduce, ameliorate, or stabilize an existing undesirable condition or its side effects).

[0029] The term "preparation" or "dosage form" is intended to include both solid and liquid formulations of the active compound and / or its crystalline solids, and one of skill in the art will appreciate that the active ingredient may be present in different formulations depending on the desired dosage and pharmacokinetic parameters.

[0030] As used herein, "excipient" refers to a compound that is used to prepare a pharmaceutical composition and is generally safe, non-toxic, biologically or otherwise undesirable, and includes excipients that are acceptable for veterinary and human pharmaceutical use.

[0031] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject without undue toxicity, irritation, allergic response, or other problem or complication, and commensurate with a reasonable benefit / risk ratio.

[0032] As used herein, the phrase "pharmaceutically acceptable carrier" means a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject.

[0033] As used herein, the phrase "co-administration" refers to any form of administration of two or more different therapeutic agents, such that the second agent is administered while the previously administered therapeutic agent is still effective in the body (e.g., the two agents are effective in the patient at the same time, which may include a synergistic effect of the two agents). For example, different therapeutic compounds may be administered either simultaneously or sequentially, either in the same formulation or in separate formulations. Thus, an individual receiving such treatment may benefit from the combined effect of the different therapeutic agents.

[0034] The recitation of a list of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of the listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0035] Whenever values ​​and ranges are provided herein, it should be understood that all values ​​and ranges encompassed by those values ​​and ranges are meant to be encompassed within the scope of the disclosure. Moreover, all values ​​that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the application.

[0036] Compounds and crystalline solids In one aspect, the present disclosure provides a crystalline solid comprising a compound of formula (I): [ka] wherein R is n-propyl. The compound of formula (I) where R is n-propyl is alternatively referred to herein as "Compound 1."

[0037] In some embodiments, the crystalline solids described herein are characterized by X-ray diffraction (XRD). In certain embodiments, the XRD is X-ray powder diffraction (XRPD). θ represents the diffraction angle and is measured in degrees. In some embodiments, the diffractometer used in XRD measures the diffraction angle as two times the diffraction angle θ. Thus, in certain embodiments, the diffraction patterns described herein refer to the X-ray intensity measured against the angle 2θ.

[0038] In some embodiments, the crystalline solid comprising the compound of formula (I) has 2θ values ​​of 16.1, 20.1, and 24.5. In some embodiments, the crystalline solid comprising the compound of formula (I) has 2θ values ​​of 16.1, 20.1, 24.5, 23.7, 18.8, 21.5, 17.7, 8.1, 9.9, 13.0, 26.3, and 30.4. In some embodiments, the crystalline solid comprising the compound of formula (I) has 2θ values ​​of 16.1, 20.1, 24.5, 23.7, 18.8, 21.5, 17.7, 8.1, 9.9, 13.0, 26.3, 30.4, 23.0, 26.6, 25.3, 25.5, and 19.7. In some embodiments, the crystalline solid comprising the compound of Formula (I) has an XRD pattern substantially as shown in Figure 1 (A or B). In some embodiments, the XRD pattern is of a methanol solvate of the compound of Formula (I). In some embodiments, the XRD pattern is of a non-solvate of the compound of Formula (I). In some embodiments, the XRD pattern corresponds to the three-dimensional shape corresponding to Figure 1C.

[0039] In certain embodiments, the compound of formula (I) is not solvated or hydrated in the crystalline solid (e.g., the crystal lattice does not contain solvent or water molecules). In certain embodiments, the crystalline solid comprising the compound of formula (I) contains non-hydrated water and / or non-solvating solvent. In certain embodiments, such non-hydrated water and / or non-solvating solvent is present in residual amounts, e.g., less than 10% by weight, or less than 5% by weight, or greater than 0% by weight but less than 1% by weight.

[0040] In certain embodiments, the compound of formula (I) is solvated by one or more solvents. In certain embodiments, the compound of formula (I) is solvated by alcohol to form an alcohol solvate, preferably methanol to form a methanol solvate. In certain embodiments, the crystalline methanol solvate of the compound of formula (I) contains about 1.0, about 1.1, or about 1.2 molecules of methanol per molecule of the compound of formula (I). In certain embodiments, the compound of formula (I) is solvated by ethanol. In certain embodiments, the compound of formula (I) is hydrated by water. In certain embodiments, the compound of formula (I) is solvated / hydrated by ethanol and water. In various embodiments, the crystalline solid is a solvate selected from a methanol solvate, an ethanol solvate, a 1-propanol solvate, a 2-propanol solvate, a C-4 alcohol solvate, a C-5 alcohol solvate, and a C-6 alcohol solvate, preferably a methanol solvate.

[0041] In various embodiments, the compound of formula (I) having the XRD pattern disclosed herein is prepared from amorphous material of greater than 90% purity, comprising dissolving the amorphous material in alcohol and precipitating the product over time, preferably at ambient temperature. In various embodiments, the compound of formula (I) is prepared from amorphous material by a method comprising dissolving the amorphous material in water or an aqueous solution, then diluting the resulting solution with a glycerol solvent and precipitating the compound of formula (I) over time. In various embodiments, the glycerol solvent is an alcohol, such as ethanol, methanol, propanol, or another alcohol of 8 carbons or less. In various embodiments, the glycerol solvent is ethanol. In various embodiments, the glycerol solvent is denatured ethanol.

[0042] In one aspect, the disclosure provides a crystalline solid comprising a compound of formula (II). [ka]

[0043] The compound of formula (II) is alternatively referred to herein as "Compound 2."

[0044] In some embodiments, the crystalline solid comprising the compound of formula (II) has 2θ values ​​of 21.5, 24.2, 26.7, and 19.6. In some embodiments, the crystalline solid comprising the compound of formula (II) has 2θ values ​​of 21.5, 24.2, 26.7, 19.6, 15.6, and 29.3. In some embodiments, the crystalline solid comprising the compound of formula (II) has 2θ values ​​of about 21.5, 24.2, 26.7, 19.6, 15.6, 29.3, 22.9, 23.1, 22.5, 13.3, 22.2, 30.0, 30.6, 13.1, 27.2, or 17.5. In some embodiments, the crystalline solid comprising the compound of formula (II) has an XRD pattern substantially as shown in FIG. 2. In some embodiments, the XRD pattern is that of a hydrate of the compound of formula (II).

[0045] In certain embodiments, the compound of formula (II) is not solvated or hydrated in the crystalline solid (e.g., the crystal lattice does not contain solvent or water molecules). In certain embodiments, the compound of formula (II) is solvated by one or more solvents. In certain embodiments, the crystalline solid comprising the compound of formula (II) contains non-hydrated water and / or non-solvating solvent. In certain embodiments, such non-hydrated water and / or non-solvating solvent is present in a residual amount, for example, less than 10% by weight, or less than 5% by weight, or greater than 0% by weight but less than 1% by weight.

[0046] In some embodiments, the compound of formula (II) is solvated with water to form a hydrate. In other embodiments, the compound of formula (II) is solvated with alcohol to form an alcohol solvate. In certain embodiments, the crystalline hydrate of the compound of formula (II) contains about 1.0, about 1.1, or about 1.2 molecules of water per molecule of the compound of formula (II). In certain embodiments, the compound of formula (II) is solvated with ethanol. In certain embodiments, the compound of formula (II) is hydrated with water. In certain embodiments, the compound of formula (II) is solvated / hydrated with ethanol and water. In various embodiments, the crystalline solid comprising the compound of formula (II) is a solvate selected from a methanol solvate, an ethanol solvate, a 1-propanol solvate, a 2-propanol solvate, a C-4 alcohol solvate, a C-5 alcohol solvate, and a C-6 alcohol solvate, preferably a methanol solvate.

[0047] In various embodiments, the compound of formula (II) having the XRD pattern disclosed herein is prepared from an amorphous material of greater than 90% purity by a method comprising dissolving the amorphous material in an alcohol and precipitating the compound of formula (II) over time, preferably at ambient temperature. In various embodiments, the compound of formula (II) is prepared from an amorphous material by a method comprising dissolving the amorphous material in water or an aqueous solution, then diluting the resulting solution with a philic solvent and precipitating the compound of formula (II) over time. In various embodiments, the philic solvent is an alcohol, such as ethanol, methanol, propanol, or another alcohol of 8 carbons or less. In various embodiments, the philic solvent is ethanol. In various embodiments, the philic solvent is denatured ethanol.

[0048] It will be apparent that the compounds of formula (I) and formula (II) may exist in various protonation states depending, inter alia, on the pH of their environment. In various pH environments, the compounds of formula (I) and formula (II) may exist as zwitterions, or inner salts, as indicated herein.

[0049] In various embodiments, the compounds of formula (I) and (II) are one or more salts, the salts being represented by H + , Li + , Na + , K + , Mg 2+ , and Ca 2+ and / or the salt is formed with a cation selected from acetate, trifluoromethanesulfonate (triflate), halide, trifluoroacetate, formate, H2PO4 - , HPO4 2- , O.H. - , HSO4 - , SO4 2- , NO3 - , HCO3 - , and CO3 2- and mixtures thereof. In various embodiments, the compound is a zwitterion.

[0050] The present disclosure includes the use of pharma- ceutically acceptable salts of the compounds of the present disclosure and / or their crystalline solids.In certain embodiments, the contemplated salts of the present disclosure include, but are not limited to, alkyl, dialkyl, trialkyl or tetraalkyl ammonium salts.In certain embodiments, the contemplated salts of the present disclosure include, but are not limited to, L-arginine, benthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine and zinc salts.

[0051] In certain embodiments, the compounds are salts having an anion selected from acetate, triflate, halide, trifluoroacetate, or formate. In other embodiments, when the disclosed compounds are in contact with a medium, e.g., an aqueous medium, the anion is, for example, OH - , H2PO4 - , HPO4 2- , HSO4 - , SO4 2- , NO3 - , HCO3 - , and CO3 2- You can choose from.

[0052] In some embodiments, the disclosed compounds are in the form of negatively charged phosphate salts, which can form salts with any suitable cation. The cation can change as the compound is isolated or transferred to a medium with a different anion species. For example, the disclosed compounds can be in the form of phosphate salts, which are pharma- ceutically acceptable salts as described herein. In certain embodiments, the cation is Li + , Na + , K + , Mg 2+ , and Ca 2+ You can choose from.

[0053] In some embodiments, the crystalline solids described herein are not part of a solution, suspension, mixture, slurry, reaction mixture, and the like.

[0054] In some embodiments, the average size of a single crystal of the crystalline solid comprising a compound of Formula (I) or Formula (II) is greater than about 1 micrometer, greater than about 5 micrometers, greater than about 10 micrometers, or greater than about 20 micrometers. In further embodiments, the average size of a single crystal of the crystalline solid comprising a compound of Formula (I) or Formula (II) is about 1 to about 100 micrometers, about 20 to about 100 micrometers, about 1 to about 500 micrometers, about 1 to about 250 micrometers, about 20 to about 250 micrometers, or about 20 to about 500 micrometers.

[0055] In some embodiments, the crystalline solids described herein have a lower solubility in water compared to the amorphous solids of the compounds of Formula (I) and Formula (II). In preferred embodiments, the solubility ratio of the crystalline solid to water is about 1:5 to about 1:75 by weight. In more preferred embodiments, the solubility ratio of the crystalline solid to water is about 1:10 to about 1:60 by weight. This lower solubility in water may impart desirable therapeutic properties.

[0056] In various embodiments, the crystalline solid is anhydrous. In various embodiments, the crystalline solid contains less than about 5% water, less than about 2% water, less than about 1% water, less than about 0.5% water, or less than about 0.1% water. In some embodiments, the percentages are by weight.

[0057] In preferred embodiments, the compound of formula (I) or its crystalline solid comprises less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. For example, in preferred embodiments, the compound of formula (I) or its crystalline solid comprises less than about 5% propyl nicotinate, less than about 2% propyl nicotinate, less than about 1% propyl nicotinate, less than about 0.5% propyl nicotinate, or less than about 0.1% propyl nicotinate. In some embodiments, the percentages are by weight.

[0058] In preferred embodiments, the compound of formula (II) or its crystalline solid comprises less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities.For example, in preferred embodiments, the compound of formula (II) or its crystalline solid comprises less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, less than about 0.1% nicotinic acid riboside, or less than about 0.01% nicotinic acid riboside.In some embodiments, percentages are by weight.

[0059] In certain preferred embodiments, the crystalline solid comprising the compound of formula (I) or formula (II) is pure or substantially pure. In certain preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably greater than about 98% pure, for example, greater than about 99% pure. In some embodiments, the percentages are by weight. In preferred embodiments, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of the compound of formula (I) or formula (II). In some embodiments, the percentages are by weight.

[0060] The crystalline solids described herein may have advantageous properties compared to amorphous forms of the compounds of Formula (I) and Formula (II). In some embodiments, the crystalline solids exhibit improved chemical and / or physical stability, for example at elevated temperatures. In certain embodiments, compositions comprising the crystalline solids exhibit improved chemical and / or physical stability. In some embodiments, the crystalline solids have improved storage stability. In certain embodiments, the crystalline solids exhibit better handling properties in manufacturing processes than the amorphous forms, which may result in compounds, crystalline solids, and compositions having higher purity, stability, and / or consistency. In some embodiments, the crystalline solids may be easier to process under typical pharmaceutical processing conditions. In certain embodiments, the improved handling properties include improved adhesion and flow properties. In some embodiments, the crystalline solids described herein are less hygroscopic compared to the amorphous forms. For example, when exposed to a humid environment (e.g., at least 50% humidity), the crystalline solids may occupy less water than the corresponding amorphous forms under the same conditions. In certain embodiments, the crystalline solids maintain structural integrity when exposed to humidity, and may be, for example, less susceptible to swelling or conversion to an unstable form. In some embodiments, the crystalline solids described herein have a lower solubility and / or dissolution rate compared to the amorphous form. In certain embodiments, it is desirable to slow the absorption of the crystalline solids to extend their effect as drugs. For example, in some embodiments, the crystalline solids described herein are effectively delivered to the intestine and do not dissolve significantly in the stomach. In some embodiments, the sustained release effect is achieved, for example, by using an aqueous suspension of the crystalline solid. In other embodiments, the delayed release is achieved by dissolving or suspending the solid material in an oil vehicle. In some embodiments, the crystalline solids described herein have a higher purity than the amorphous form and / or facilitate large-scale preparation of the pure material, for example, at lower cost or with less material or space-intensive purification methods.In some such embodiments, the crystalline solids and methods described herein facilitate large-scale purification, for example, purification of greater than about 1 gram, greater than about 10 grams, or greater than about 100 grams.

[0061] Method for preparing a crystalline solid Also provided herein is a method for preparing a crystalline solid of a compound of formula (I). In certain embodiments, the present disclosure relates to a method for preparing a crystalline solid of a compound of formula (I), comprising: a) dissolving a compound of formula (I) in a solvent to form a mixture; and b) crystallizing the compound of formula (I) from the mixture to form a crystalline solid.

[0062] In a preferred embodiment, the mixture comprising the compound of formula (I) is a solution. In other embodiments, the mixture is a slurry or suspension. In some embodiments, the solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, and other liquids with similar polarity and properties, and combinations thereof. In some embodiments, the solvent comprises acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMSO), methanol, ethanol, ethyl acetate, isopropyl acetate, methanol, methyl ethyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, propanol, butanol, water, or any combination thereof. In certain embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including branched and unbranched isomers thereof. In a preferred embodiment, the solvent is methanol. In some embodiments, the solvent comprises two or more of the solvents described herein. In some embodiments, the solvent is anhydrous.

[0063] In certain embodiments, the temperature of the solvent is above ambient temperature during the dissolving step. In such embodiments, the method includes heating the solvent. For example, the temperature of the solvent can be about 30 to about 50° C. or about 30 to about 40° C., for example about 35° C. In other embodiments, the temperature of the solvent is about ambient temperature during the dissolving step. In yet other embodiments, the temperature of the solvent is below ambient temperature during the dissolving step. In such embodiments, the method includes cooling the solvent. In some embodiments, the temperature of the solvent is about 20 to about 30° C., for example about 25° C. In a preferred embodiment, the compound of formula (I) is completely dissolved in the solvent before the crystallization step. By completely dissolved, it is meant that the compound results in a homogenous solution, not a slurry or suspension. In other embodiments, the compound of formula (I) is partially dissolved in the solvent before the crystallization step.

[0064] In certain embodiments, the method includes forming a supersaturated solution from a mixture (e.g., a solution) of a compound of Formula (I), where the supersaturated solution is supersaturated with respect to the compound of Formula (I). In some embodiments, the supersaturated solution has a supersaturation ratio of about 1 to about 4, e.g., about 2. In some such embodiments, the compound of Formula (I) is precipitated (e.g., crystallized) from the supersaturated solution. In some embodiments, the resulting precipitate (e.g., crystals) is a crystalline solid as described herein.

[0065] The supersaturated solution may be formed according to a variety of methods. In some embodiments, forming a supersaturated solution may include adding a phagocytic solvent to a mixture (e.g., a solution), reducing the temperature of the mixture (e.g., a solution), reducing the volume of the mixture (e.g., a solution), or any combination thereof. For example, a method may include adding a phagocytic solvent, followed by cooling the resulting mixture, followed by adding additional phagocytic solvent.

[0066] In certain embodiments, forming a supersaturated solution includes lowering the temperature of a mixture containing a compound of Formula (I). In some such embodiments, the temperature of the solution is lowered to about 0 to about 25° C., about 0 to about 10° C., or about −5 to about 5° C., for example, to about 0° C. In certain embodiments, cooling the solution can be passive (e.g., allowing the solution to stand at ambient temperature) or active (e.g., cooling the solution in an ice bath or freezer).

[0067] In some embodiments, forming a supersaturated solution includes adding a soluble solvent to a mixture containing the compound of formula (I). As used herein, "soluble solvent" means a liquid in which the compounds of formula (I) and (II) are insoluble, minimally soluble, or partially soluble. In effect, adding a soluble solvent to a solution in which the compounds of formula (I) and (II) are dissolved reduces the solubility of the compounds of formula (I) and (II) in the solvent, thereby stimulating precipitation.

[0068] In certain embodiments, the phasic solvent may be added slowly to prevent uncontrolled crystallization. In some embodiments, the phasic solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, and other liquids with similar polarity and properties that are miscible with the solvent, and combinations thereof. In some embodiments, the phasic solvent is an alkane solvent, such as hexane or pentane solvent, or an aromatic hydrocarbon solvent, such as benzene, toluene, or xylene. In certain embodiments, the phasic solvent is selected from ethyl acetate, isopropyl acetate, methyl tert-butyl ether, methyl isobutyl ketone, tetrahydrofuran, 1-propanol, 2-propanol, ethanol, denatured ethanol, and combinations thereof. In a preferred embodiment, the phasic solvent is TBME. In some embodiments, the phasic solvent comprises two or more of the solvents described herein. In some embodiments, the ratio of solvent to phasic solvent is about 1:1 to about 8:1 by volume, or about 4:1 to about 6:1 by volume, such as about 5:1 by volume.

[0069] In certain embodiments, the method further comprises evaporating the solvent from the mixture, hi some embodiments, the solvent may be removed under reduced pressure and / or by heating the solvent such that it evaporates.

[0070] In certain embodiments, crystallization comprises secondary nucleation. In some embodiments, crystallization comprises adding seed crystals to the solution, and the seed crystals comprise a compound of formula (I). In certain embodiments, the seed crystals are formed during pre-crystallization. In certain such embodiments, the pre-crystallization is carried out on a smaller scale than the crystallization to which the seed crystals are added.

[0071] In other embodiments, secondary nucleation may be caused by other changes to the mixture's environment. For example, crystallization may be promoted by environmental changes including, but not limited to, a crystallization wall, an agitation impeller, and ultrasonic treatment.

[0072] In a preferred embodiment, the method includes isolating the crystalline solid, for example by filtering the crystals, by decanting the fluid from the crystals, or by any other suitable separation technique.

[0073] In certain embodiments, the method includes washing the crystalline solid comprising the compound of formula (II), for example, washing the crystalline solid with a solvent or a mixture of one or more of the solvents and / or soluble solvents described herein. In certain embodiments, washing the crystalline solid includes washing with a liquid selected from an soluble solvent, a solvent, an alcohol, a ketone, a carboxylic acid, an ester, an ether, an alkane, water, an amine, other liquids of similar polarity and properties, and combinations thereof. In some embodiments, the liquid is selected from acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMSO), ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, an alcohol such as methanol, ethanol, propanol, or butanol, water, an alkane solvent such as pentane, hexane, or heptane, an aromatic hydrocarbon solvent such as benzene, toluene, or xylene, methyl tert-butyl ether, and combinations thereof. In certain embodiments, the solvent and / or soluble solvent is cooled before washing. In some embodiments, the method includes drying the crystalline solid, for example, under reduced pressure and / or by heating the crystalline solid and / or under a flow of drying gas, such as nitrogen, argon, or air.

[0074] In certain embodiments, the method of making the crystalline solid removes one or more impurities from the compound of formula (I). In some embodiments, the method does not include chromatography or lyophilization to purify the compound of formula (I). In certain such embodiments, the methods described herein are used to purify the compound of formula (I), for example, as a final purification step in the manufacture of the compound of formula (I).

[0075] The method described herein may provide the advantage of, among others, removing impurities from the compound of formula (I).In preferred embodiments, the crystalline solid comprises less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities.In some preferred embodiments, the crystalline solid comprises less than about 5% propyl nicotinate, less than about 2% propyl nicotinate, less than about 1% propyl nicotinate, less than about 0.5% propyl nicotinate, or less than about 0.1% propyl nicotinate.In some embodiments, the percentages are by weight.

[0076] In certain preferred embodiments, the crystalline solid comprising the compound of formula (II) is pure or substantially pure. In certain preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably, greater than about 98% pure. In some embodiments, the percentages are by weight.

[0077] In preferred embodiments, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of the compound of Formula (I). In some embodiments, the percentages are by weight.

[0078] Another aspect of the present disclosure provides a method for preparing a crystalline solid of a compound of formula (II).In certain embodiments, the present disclosure provides a method for preparing a crystalline solid of a compound of formula (II), comprising: a) dissolving a compound of formula (II) in a solvent to form a mixture; and b) crystallizing the compound of formula (II) from the mixture to form a crystalline solid.In certain embodiments, the method comprises reacting nicotinic acid riboside with a phosphorus-containing group (e.g., phosphorus oxychloride) to provide a compound of formula (II) prior to the dissolving and crystallizing steps.

[0079] In a preferred embodiment, the mixture comprising the compound of formula (II) is a solution. In other embodiments, the mixture is a slurry or suspension. In some embodiments, the solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, and other liquids with similar polarity and properties, and combinations thereof. In some embodiments, the solvent comprises acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMSO), methanol, ethanol, ethyl acetate, isopropyl acetate, methanol, methyl ethyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, propanol, butanol, water, or any combination thereof. In certain embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including branched and unbranched isomers thereof. In a preferred embodiment, the solvent is water. In certain embodiments, the solvent comprises an alcohol, such as 1-propanol. In some embodiments, the solvent comprises two or more of the solvents described herein. In some embodiments, the ratio of the compound of Formula (II) to the solvent is from about 1:2 to about 1:4 by weight, such as about 1:3 by weight.

[0080] In certain embodiments, the temperature of the solvent is above ambient temperature during the dissolving step. In such embodiments, the method includes heating the solvent. For example, the temperature of the solvent can be about 30 to about 50° C. or about 30 to about 40° C., for example about 35° C. In other embodiments, the temperature of the solvent is about ambient temperature during the dissolving step. In yet other embodiments, the temperature of the solvent is below ambient temperature during the dissolving step. In such embodiments, the method includes cooling the solvent. In some embodiments, the temperature of the solvent is about 20 to about 30° C., for example about 25° C. In a preferred embodiment, the compound of formula (II) is completely dissolved in the solvent before the crystallization step. By completely dissolved, it is meant that the compound occurs in a homogenous solution, not a slurry or suspension. In other embodiments, the compound of formula (II) is partially dissolved in the solvent before the crystallization step.

[0081] In certain embodiments, the method includes forming a supersaturated solution from a mixture (e.g., a solution) of a compound of Formula (II), where the supersaturated solution is supersaturated with respect to the compound of Formula (II). In some embodiments, the supersaturated solution has a supersaturation ratio of 1 to about 4, e.g., about 2. In some such embodiments, the compound of Formula (I) is precipitated (e.g., crystallized) from the supersaturated solution. In some embodiments, the resulting precipitate (e.g., crystals) is a crystalline solid as described herein.

[0082] The supersaturated solution may be formed according to a variety of methods. In some embodiments, forming a supersaturated solution may include adding a phagocytic solvent to a mixture (e.g., a solution), reducing the temperature of the mixture (e.g., a solution), reducing the volume of the mixture (e.g., a solution), or any combination thereof. For example, a method may include adding a phagocytic solvent, followed by cooling the resulting mixture, followed by adding additional phagocytic solvent.

[0083] In certain embodiments, forming a supersaturated solution includes lowering the temperature of a mixture containing the compound of formula (II). In some such embodiments, the temperature of the solution is lowered to about 0 to about 25° C., about 0 to about 10° C., or about −5 to about 5° C., for example, to about 0° C. In certain embodiments, cooling the solution can be passive (e.g., allowing the solution to stand at ambient temperature) or active (e.g., cooling the solution in an ice bath or freezer).

[0084] In some embodiments, forming a supersaturated solution includes adding a philic solvent to a mixture including the compound of formula (II). In certain embodiments, the philic solvent may be added slowly to prevent uncontrolled crystallization. In some embodiments, the philic solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, and other liquids with similar polarity and properties that are miscible with the solvent, and combinations thereof. In some embodiments, the philic solvent is an alkane solvent, such as hexane or pentane solvent, or an aromatic hydrocarbon solvent, such as benzene, toluene, or xylene. In certain embodiments, the philic solvent is selected from ethyl acetate, isopropyl acetate, methyl tert-butyl ether, methyl isobutyl ketone, tetrahydrofuran, and combinations thereof. In other embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including branched and unbranched isomers thereof. In a preferred embodiment, the philic solvent is 1-propanol. In some embodiments, the philic solvent includes two or more of the solvents described herein. In some embodiments, the ratio of solvent to soluble solvent is from about 1:0 to about 1:2 by volume, or from about 1:0 to about 6:7 by volume, such as about 6:7 by volume. In certain embodiments, the supersaturated solution is formed without the addition of a soluble solvent.

[0085] In certain embodiments, the method further comprises evaporating the solvent from the mixture, hi some embodiments, the solvent may be removed under reduced pressure and / or by heating the solvent such that it evaporates.

[0086] In some embodiments, the method further comprises adding an acid or base to adjust the pH of the mixture (e.g., the solution) to change the protonation state of the compound of formula (II). In certain embodiments, the base is an amine, such as triethylamine. In some embodiments, the method further comprises adding a base to the solution. In certain embodiments, the pH of the solution is adjusted to about 2 to about 4, e.g., about 3.

[0087] In certain embodiments, the crystallization comprises secondary nucleation. In some embodiments, the crystallization comprises adding seed crystals to the solution, the seed crystals comprising a compound of formula (II). In certain embodiments, the seed crystals are formed during pre-crystallization. In certain such embodiments, the pre-crystallization is carried out on a smaller scale than the crystallization to which the seed crystals are added.

[0088] In other embodiments, secondary nucleation may be caused by other changes to the mixture's environment. For example, crystallization may be promoted by environmental changes including, but not limited to, a crystallization wall, an agitation impeller, and ultrasonic treatment.

[0089] In a preferred embodiment, the method includes isolating the crystalline solid, for example by filtering the crystals, by decanting the fluid from the crystals, or by any other suitable separation technique.

[0090] In certain embodiments, the method includes washing the crystalline solid comprising the compound of formula (II), for example, washing the crystalline solid with a solvent or a mixture of one or more of the solvents and / or solvents described herein. In certain embodiments, washing the crystalline solid includes washing with a liquid selected from a solvent, a solvent, an alcohol, a ketone, a carboxylic acid, an ester, an ether, an alkane, water, an amine, other liquids of similar polarity and properties, and combinations thereof. In some embodiments, the liquid is selected from acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethylsulfoxide (DMSO), ethyl acetate, isopropyl acetate, methyl ethyl ketone, methyl isobutyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, an alcohol such as methanol, ethanol, propanol, or butanol, water, an alkane solvent such as pentane, hexane, or heptane, an aromatic hydrocarbon solvent such as benzene, toluene, or xylene, methyl tert-butyl ether, and combinations thereof. In a preferred embodiment, the method includes washing the crystalline solid with a 2:1 by volume mixture of 1-propanol and water, optionally followed by washing the crystalline solid with MTBE. In certain embodiments, the solvent and / or solvent is cooled prior to washing. In some embodiments, the method includes drying the crystalline solid, for example, under reduced pressure and / or by heating the crystalline solid.

[0091] In certain embodiments, the method of making the crystalline solid removes one or more impurities from the compound of formula (II). In some embodiments, the method does not include chromatography or lyophilization to purify the compound of formula (II). In certain such embodiments, the method described herein is used to purify the compound of formula (II), for example, as a final purification step in the manufacture of the compound of formula (II).

[0092] The method described herein may provide the advantage of, among others, removing impurities from the compound of formula (II).In preferred embodiments, the crystalline solid comprises less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities.In some preferred embodiments, the crystalline solid comprises less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, or less than about 0.1% nicotinic acid riboside.In some embodiments, the percentages are by weight.

[0093] In certain preferred embodiments, the crystalline solid comprising the compound of formula (II) is pure or substantially pure. In certain preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably, greater than about 98% pure. In some embodiments, the percentages are by weight.

[0094] In preferred embodiments, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of the compound of Formula (II). In some embodiments, the percentages are by weight.

[0095] synthesis In various embodiments, the disclosure provides a method of forming a compound of formula I, [ka] wherein R is C1-C6 alkyl or C2-C6 alkenyl, and the process comprises reacting a compound of formula (II) with [ka] This involves contacting with an alcohol R-OH in the presence of an acid. See, for example, Scheme 1. [ka]

[0096] In certain embodiments, R is C1-C6 alkyl. In some embodiments, R is C1-C4 alkyl or C2-C4 alkenyl. In certain embodiments, R is C3 alkyl. In certain embodiments, R is n-propyl.

[0097] In some embodiments, the acid is a strong acid. In some embodiments, the acid is an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. In other embodiments, the acid is an organic acid, such as glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, salicylic acid, muconic acid, etc. In a preferred embodiment, the acid is HCl.

[0098] In some embodiments, the method includes purifying the compound of formula (II). In some embodiments, the method includes removing ammonium salts, such as triethylammonium salts, from the compound of formula (II). In certain embodiments, the compound of formula (II) is provided as a crystalline solid. In some embodiments, the method includes crystallizing the compound of formula (II) according to the methods described herein.

[0099] In some embodiments, the method includes adding a solvent to the compound of formula (II) to form a mixture, such as a solution. In some embodiments, the solvent is a polar solvent. Polar solvents include polar groups that may be selected from, for example, hydroxyl, carbonyl, ether, ester, amine, amide, and carboxyl groups. In some embodiments, the solvent includes water. In a preferred embodiment, the alcohol R-OH is the reaction solvent. In a preferred embodiment, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including branched and unbranched isomers thereof. In a more preferred embodiment, the solvent is a propanol, such as 1-propanol or 2-propanol. In some embodiments, the temperature of the mixture including the compound of formula (II) is about -5 to about 10°C, about -5 to about 5°C, or about 0°C.

[0100] In some embodiments, the method includes mixing a compound of formula (II) with an alcohol. In certain embodiments, the mixing step is carried out for about 12 to about 72 hours, about 12 to about 48 hours, or about 12 to about 24 hours. In some embodiments, the mixture containing the compound of formula (II) is at about -5 to about 10°C, about -5 to about 5°C, or about 0°C throughout the mixing step.

[0101] In some embodiments, the method includes adding a base. In various embodiments, the base is added after about 12 to about 72 hours, about 12 to about 48 hours, or about 12 to about 24 hours. In some embodiments, the base is added until the pH of the reaction mixture is about 4 to about 5. In certain such embodiments, the agent is an amine base. In certain such embodiments, the base is a trialkylamine base. In a preferred embodiment, the base is triethylamine. In further embodiments, the method includes adding seed crystals of a compound of Formula (I) to the reaction mixture.

[0102] In various embodiments, the method includes purifying the resulting product (i.e., the compound of formula (I)). In some embodiments, purifying the product includes chromatography. In other embodiments, purifying the product does not include chromatography. In a preferred embodiment, purifying the product includes crystallizing the compound of formula (I) according to the methods described herein. In a preferred embodiment, the compound of formula (I) is provided as a crystalline solid as described herein.

[0103] Treatments, Diseases, Disorders, and Conditions Provided herein are methods of regulating NAD levels in a subject in need thereof, comprising administering a compound, crystalline solid, and / or composition described herein. Any compound, crystalline solid, or composition described herein can be used in the manufacture of a medicament for the treatment of any disease or condition disclosed herein.

[0104] Provided herein are methods of treating a disease or disorder associated with NAD biosynthesis comprising administering a compound, crystalline solid, and / or composition described herein.

[0105] Methods for using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof are provided herein. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be useful for a variety of therapeutic applications, including treating and / or alleviating a wide variety of diseases and disorders, including, for example, diseases or disorders associated with aging or stress, diabetes, obesity, neurodegenerative diseases, ataxia and related muscle disorders, acute organ failure, viral symptoms such as cytokine storm, cardiovascular disease, blood clotting disorders, inflammation, cancer, and / or flushing, and the like. The methods include administering the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof to a subject in need thereof. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be useful for increasing or maintaining NAD levels in certain tissues or cells while decreasing NAD levels in other tissues or cells. In various embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to selectively decrease NAD levels in some tissues or cells, but to a lesser extent in other tissues or cells.

[0106] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can also be used to treat diseases or disorders associated with inflammation.Exemplary inflammatory conditions include, for example, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, degenerative joint disease, spondyloarthropathies, gouty arthritis, systemic lupus erythematosus, juvenile arthritis, rheumatoid arthritis, osteoarthritis, osteoporosis, diabetes (e.g., insulin-dependent diabetes mellitus or juvenile diabetes mellitus), menstrual cramps, cystic fibrosis, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, mucous colitis, ulcerative colitis, gastritis, Examples of inflammatory conditions include esophagitis, pancreatitis, peritonitis, Alzheimer's disease, shock, ankylosing spondylitis, gastritis, conjunctivitis, pancreatitis (acute or chronic), multiple organ injury syndrome (e.g., secondary to sepsis or trauma), myocardial infarction, arteriosclerosis, stroke, reperfusion injury (e.g., due to cardiopulmonary bypass or renal dialysis), acute glomerulonephritis, vasculitis, thermal injury (i.e., sunburn), necrotizing enterocolitis, granulocyte transfusion associated syndrome, and Sjogren's syndrome. Exemplary inflammatory conditions of the skin include, for example, eczema, atopic dermatitis, contact dermatitis, urticaria, scleroderma, psoriasis, and skin diseases with an acute inflammatory component.

[0107] In other embodiments, the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof can be used to treat skin conditions.Exemplary skin conditions that can be treated according to the methods described herein include disorders or diseases associated with or caused by inflammation, sun damage, or natural aging.For example, the compositions find utility in treating contact dermatitis (including irritant contact dermatitis and allergic contact dermatitis), atopic dermatitis (also known as allergic eczema), actinic keratosis, keratinopathy (including eczema), bullous epidermolysis, exfoliative dermatitis, seborrheic dermatitis, erythema (including erythema multiforme and erythema nodosum), damage caused by the sun or other light sources, discoid lupus erythematosus, dermatomyositis, psoriasis, skin cancer, and the effects of natural aging. In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat wounds and / or burns to promote healing, including, for example, first, second, or third degree burns and / or thermal, chemical, or electrical burns.

[0108] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can also be administered to subjects suffering from acute diseases, such as injury to an organ or tissue, such as subjects suffering from stroke or myocardial infarction, subjects suffering from spinal cord injury, or subjects undergoing transplantation of a solid organ, such as a liver or kidney. In some embodiments, the compounds, crystalline solids, and pharmaceutical compositions thereof can be administered to subjects suffering from acute kidney injury (AKI), also known as acute renal failure (ARF). Subjects suffering from AKI or at risk of suffering from AKI can be screened for renal function, for example, by testing for abnormal levels of serum creatinine. Subjects may be treated prophylactically or in response to acute kidney injury, such as stage 1 AKI. Subjects undergoing solid organ transplantation may be treated prophylactically or post-transplantation as a form of organ preservation, or individual organs may be treated ex vivo prior to transplantation. Subjects undergoing surgery other than organ transplantation, such as biopsy, resection, or repair of traumatic injury, can be treated prophylactically or post-operatively.

[0109] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may also be used in subjects suffering from or likely to suffer from chronic damage or chronic disease of solid organs such as the kidney or liver. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to subjects suffering from chronic kidney disease, such as end-stage renal failure, or nephropathy, or diabetic nephropathy. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to subjects suffering from chronic liver disease, such as chronic infection, cirrhosis, or liver cancer, to repair or limit further damage to the liver. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to repair alcoholic liver or stabilize or repair damage from nonalcoholic steatohepatitis (NASH) or nonalcoholic fatty liver disease (NAFLD).

[0110] In certain embodiments, the compounds, crystalline solids, or pharmaceutical compositions disclosed herein may be used to treat or prevent diseases or conditions in a subject that are caused or aggravated by cellular senescence, for example, after the onset of senescence, in methods for reducing the rate of aging in a subject, for extending the lifespan of a subject, for treating or preventing diseases or conditions associated with lifespan, for treating or preventing diseases or conditions associated with the proliferative capacity of cells, and for treating or preventing diseases or conditions resulting from cell damage or death. In certain embodiments, the methods do not act by reducing the incidence of diseases that shorten the lifespan of a subject. In certain embodiments, the methods do not act by reducing the lethality caused by a disease, such as cancer.

[0111] In certain embodiments, the compounds, crystalline solids, or pharmaceutical compositions disclosed herein may be administered to a subject to generally extend the lifespan of its cells and protect them from stress and / or apoptosis. Treating a subject with the compounds or crystalline solids described herein may be similar to subjecting the subject to hormesis, i.e., mild stress that may be beneficial to the organism and extend its lifespan.

[0112] In other embodiments, provided herein are methods for treating cardiovascular disease by administering the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof to a subject in need thereof. Cardiovascular diseases that can be treated using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include cardiomyopathies or myocarditis, such as idiopathic cardiomyopathy, metabolic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy. Also treatable using the compositions and methods described herein are atherosclerotic disorders of large blood vessels, such as the aorta, coronary arteries, carotid arteries, cerebrovascular arteries, renal arteries, iliac arteries, femoral arteries, and popliteal arteries (macrovascular diseases). Other vascular diseases that can be treated include those associated with platelet aggregation, retinal arteries, glomerular arteries, neurovascular arteries, cardiac arteries, and associated capillary beds of the eye, kidney, heart, and central and peripheral nervous system. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may also be used to increase HDL levels in the plasma of an individual.

[0113] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be administered to subjects who have recently received or are about to receive a dose of radiation or toxins. In one embodiment, the dose of radiation or toxins is received as part of a work-related or medical procedure, such as working in a nuclear power plant, flying an airplane, administering radioactive dyes for X-rays, CAT scans, or medical imaging. In such an embodiment, the compounds or crystalline solids are administered as a preventative measure. In other embodiments, radiation or toxin exposure is received unintentionally, for example, as a result of an industrial accident, living in a place of natural radiation, an act of terrorism, or an act of war involving radioactive or toxic materials. In such cases, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof are preferably administered as soon as possible after exposure to inhibit apoptosis and the subsequent development of acute radiation syndrome.

[0114] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be useful for treating age-related disorders, such as cancer. Exemplary cancers that may be treated using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include hormone-dependent cancers, including brain and kidney cancer, breast cancer, prostate cancer, testicular cancer, and ovarian cancer, lymphoma, and leukemia. Other diseases that may be treated include autoimmune diseases in which autoimmune cells should be removed, such as systemic lupus erythematosus, scleroderma, and arthritis.

[0115] Viral infections, such as herpes, HIV, adenovirus, and HTLV-1 associated malignant and benign diseases, can also be treated by administration of the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof.

[0116] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to treat patients suffering from infectious diseases such as COVID-19 and other viral infections, including those experiencing symptoms such as cytokine release syndrome (cytokine storm). In some embodiments, the compounds, crystalline solids, and pharmaceutical compositions thereof mitigate or prevent cytokine storm without necessarily treating the underlying viral infection (e.g., COVID-19). Cytokine release syndrome is an acute systemic inflammatory syndrome that can result from a variety of causes. In particular, cytokine storm has been described in COVID-19 and other severe viral syndromes (SARS, MERS). A subset of patients exhibits significantly elevated cytokines, and severely ill patients can also exhibit much higher levels of IL6, CRP, ferritin, D-dimer, and other markers, as well as lymphopenia (reduced numbers of CD4+ and CD8+ T cells). For example, one report identified a subset of patients with a higher likelihood of death when D-dimer levels at admission were greater than 2.0ug / ml (12 / 67>=2.0 vs. 1 / 267<2.0, sensitivity 92.3%, specificity 83.3%) ("D-dimer levels on admission to predict in-hospital mortality in patients with Covid-19." Zhang L, Yan X, Fan Q, et al. J Thromb Haemost. 2020 Apr 19). NAD regulates the NLRP3 inflammasome release of IL-1β, which may regulate cytokine storm. NAD levels are known to decline with age, which may also contribute to worse outcomes in older COVID-19 patients. One aspect of the present disclosure provides a method of treating COVID-19 in a human patient, comprising administering to the patient a compound, crystalline solid, and pharmaceutical composition thereof of the present disclosure in the absence of administration of zinc sulfate, betaine, or mixtures thereof.

[0117] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to treat patients suffering from neurodegenerative diseases and traumatic or mechanical damage to the central nervous system (CNS) or peripheral nervous system (PNS). Examples of neurodegenerative diseases include, but are not limited to, ataxia, Alzheimer's disease (AD), non-Alzheimer's dementia, Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS, Lou Gehrig's disease), diffuse Lewy body disease, acanthocytic chorea, primary lateral sclerosis, multiple sclerosis (MS), eye disease (ophthalmic neuritis), spinal muscular atrophy, chemotherapy-induced neuropathy (e.g., from vincristine, paclitaxel, bortezomib), diabetes-induced neuropathy, and Friedreich's ataxia.

[0118] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat skeletal muscle disorders, muscle disorders, and conditions involving muscle loss, atrophy, and sarcopenia.

[0119] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to reduce appetite and / or increase satiety, thereby causing weight loss or avoidance of weight gain. A subject in need of such treatment can be a subject who is overweight, obese, or at high risk of becoming overweight or obese.

[0120] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat subjects who have or may develop cachexia. The method may further include monitoring the disease state of the subject. The method for promoting appetite and / or weight gain may include, for example, pre-identifying a subject in need of reduced fat or lipid metabolism, for example, by measuring the subject's weight and determining the subject's BMI. The method may also include monitoring the subject, for example, during and / or after administration of the disclosed compounds, crystalline solids, or pharmaceutical compositions thereof. Administration may include, for example, one or more doses delivered bolus or continuously. Monitoring may include evaluating hormones or metabolites. Exemplary hormones include leptin, adiponectin, resistin, and insulin. Exemplary metabolites include triglycerides, cholesterol, and fatty acids.

[0121] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to treat metabolic disorders such as insulin resistance, prediabetes, type II diabetes, and / or complications thereof. Administration of the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can increase insulin sensitivity and / or decrease insulin levels in a subject. A subject in need of such treatment can be a subject with insulin resistance or other precursors to type II diabetes, a subject with type II diabetes, or a subject likely to develop any of these conditions. For example, a subject can be a subject with insulin resistance, e.g., a subject with high circulating levels of insulin, and / or a subject with an associated condition, e.g., hyperlipidemia, dyslipidemia, hypercholesterolemia, impaired glucose tolerance, high blood glucose levels, other symptoms of syndrome X, hypertension, atherosclerosis, and lipodystrophy.

[0122] Provided herein is a process for regulating blood glucose concentration in a mammal. As used herein, regulating blood glucose concentration refers to any increase, decrease, and / or maintenance in or of blood glucose concentration compared to a previously determined level.

[0123] Therapeutic methods disclosed herein are also directed to methods of modulating the circadian clock, thereby regulating or affecting biological functions that are modulated (sometimes also referred to as influenced, associated, or mediated) by activity of the circadian clock. Typically, these biological functions exhibit patterns of activity and inactivity that are generally repeated approximately every 24 hours and oscillate between "active" and "inactive" states over the course of a 24-hour period.

[0124] Thus, the present disclosure provides a method of regulating the activity of the circadian clock by administering a compound, crystalline solid, or pharmaceutical composition disclosed herein to a mammal in need thereof. Generally, the regulation of the activity of the circadian clock is the result of regulating CLOCK:BMAL1, which is achieved according to the present method by regulating the activity of SIRT1. The activity of SIRT1 is generally regulated according to the present method by administering a compound, crystalline solid, or pharmaceutical composition disclosed herein, and in certain embodiments, by administering a compound or crystalline solid that affects the NAD pathway. The regulation of the circadian clock thereby allows for the regulation of activities mediated by the circadian clock.

[0125] According to the present disclosure, the activity of the circadian clock can be increased, decreased, or maintained by administration of the compounds, crystalline solids, or pharmaceutical compositions disclosed herein. Accordingly, the biological functions (sometimes referred to as biological activities) regulated by the activity of the circadian clock can also be increased, decreased, or maintained. In addition, these biological functions can also be time-shifted, i.e., activities that typically occur during a certain period, such as daytime or daytime hours (sometimes referred to as light cycle), or nighttime or nighttime hours (sometimes referred to as dark cycle), can instead be shifted so that they occur during the dark cycle or light cycle, respectively.

[0126] In various embodiments, disclosed herein are methods of differentially regulating nicotinamide adenine dinucleotide (NAD) levels in two or more tissues or cell types. Such methods can include administering a compound, crystalline solid, or composition disclosed herein, which administration induces a differential response in NAD levels in a first tissue or cell type compared to a second tissue or cell type. In various embodiments, the differential response in NAD levels is selected from at least a 10% difference in NAD levels, at least a 20% difference in NAD levels, at least a 30% difference in NAD levels, at least a 40% difference in NAD levels, at least a 50% difference in NAD levels, at least a 60% difference in NAD levels, at least a 70% difference in NAD levels, at least an 80% difference in NAD levels, at least a 90% difference in NAD levels, at least a 100% difference in NAD levels, at least a 200% difference in NAD levels, at least a 300% difference in NAD levels, at least a 400% difference in NAD levels, at least a 500% difference in NAD levels, at least a 600% difference in NAD levels, at least a 700% difference in NAD levels, at least an 800% difference in NAD levels, at least a 900% difference in NAD levels, and at least a 1000% difference in NAD levels. In various embodiments, the differential response in NAD levels is at least a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% increase in NAD levels in the first tissue or cell type compared to the untreated NAD level or NAD level before treatment. The increase in AD levels and a concomitant decrease in NAD levels of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% in the second tissue or cell type compared to untreated NAD levels or NAD levels before treatment.In various embodiments, the NAD level differential response is maintenance of NAD levels within 10% in said first tissue or cell type compared to untreated NAD levels, and a concomitant decrease in NAD levels of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% in a second tissue or cell type compared to untreated NAD levels. In various embodiments, the differential response in NAD levels is a decrease in NAD levels in a first tissue or cell type of at least 10% compared to untreated NAD levels, and a concomitant decrease in NAD levels in a second tissue or cell type compared to untreated NAD levels, where the decrease in the second tissue or cell type is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% less than the decrease in the first tissue or cell type. In various embodiments, the first tissue or cell type is a normal tissue or cell and the second tissue or cell type is neoplastic or cancerous.

[0127] The methods of treating cancer disclosed herein include treating an individual in need thereof. Exemplary cancers that may be treated using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include brain and kidney cancer, hormone-dependent cancers including breast cancer, prostate cancer, testicular cancer, and ovarian cancer, lymphoma, and leukemia. In various embodiments, the cancer may be a common type of cancer in men, such as lung cancer, prostate cancer, colorectal cancer, and gastric cancer. In various embodiments, the cancer may be a common type of cancer in women, such as breast cancer, colorectal cancer, lung cancer, and cervical cancer. In various embodiments, the cancer may be a skin cancer, such as melanoma, squamous cell carcinoma, or basal cell carcinoma. In various embodiments, the cancer may be a common type of cancer in children, such as acute lymphoblastic leukemia, brain cancer, or non-Hodgkin's lymphoma. In various embodiments, the methods exhibit selective cytostatic or cytotoxic effects, which are demonstrated by a decrease in the viability of the neoplastic or cancerous tissue or cells compared to untreated neoplastic or cancerous tissue or cells.

[0128] The method includes the situation where the first tissue or cell type is a normal tissue or cell, and the method is a treatment for promoting the health or increasing the biological activity of the first tissue or cell type in an individual in need thereof. In various embodiments, the treatment does not induce an increased risk of cancer diagnosis in the treated individual. Preferably, the treatment reduces the risk of cancer diagnosis in the individual receiving the treatment.

[0129] Various methods include treating or inhibiting cancer in an individual in need thereof, the methods comprising administering a compound, crystalline solid, or composition described herein. In various embodiments, disclosed herein are methods of increasing or maintaining healthy tissue or cells in an individual in need thereof without increasing the risk of neoplastic or cancerous tissue or cell growth, such methods comprising administering a compound, crystalline solid, or composition described herein.

[0130] In various embodiments, described herein are methods of increasing or maintaining healthy tissues or cells in an individual in need thereof while inhibiting the growth of neoplastic or cancerous tissues or cells, such methods comprising administering a compound, crystalline solid, or composition described herein. In various embodiments, disclosed methods include methods of increasing or maintaining nicotinamide adenine dinucleotide (NAD) levels in at least one healthy tissue or cell type, such methods comprising administering a compound, crystalline solid, or composition described herein to the healthy tissue or cell type. In various embodiments, described herein are methods of decreasing the viability of at least one cancerous tissue or cell type, such methods comprising administering a compound, crystalline solid, or composition described herein to the cancerous tissue or cell type.

[0131] In addition, the methods described herein include methods of modulating the level of NAD in at least one tissue or cell type in a mixture of tissues or cell types, such methods including targeted delivery of a compound, crystalline solid, or composition described herein to a desired tissue or cell type. In various embodiments, the targeted delivery is non-systemic.

[0132] Compositions and pharmaceutical compositions Compositions of the disclosed compounds and crystalline solids are also provided herein. In certain embodiments, the compositions include 1) a crystalline solid comprising a compound of formula (I) or formula (II) or a salt thereof, and 2) one or more pharma- ceutically acceptable excipients. In other embodiments, the compositions include 1) a compound of formula (I) or formula (II) or a salt thereof, and 2) one or more pharma-ceutically acceptable excipients.

[0133] In some embodiments, the composition is a solution. For example, in some embodiments, a crystalline solid comprising a compound of formula (I) or formula (II) is dissolved in a solvent or carrier to form a solution of the compound of formula (I) or formula (II). In preferred embodiments, the crystalline solid is of such a purity that the resulting solution is pure or substantially pure and / or free or substantially free of one or more impurities.

[0134] In certain preferred embodiments, the present disclosure provides a composition comprising a compound of formula (I) or formula (II), or a crystalline solid comprising a compound of formula (I) or formula (II), wherein the composition is pure or substantially pure. In certain preferred embodiments, the composition is greater than about 90% pure. More preferably, the composition is greater than about 95% pure, or even more preferably greater than about 98% pure, for example, greater than about 98% pure. In some embodiments, the percentages are by weight.

[0135] In a preferred embodiment, the composition comprises less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. For example, in a preferred embodiment, the composition comprises less than about 5% propyl nicotinate, less than about 2% propyl nicotinate, less than about 1% propyl nicotinate, less than about 0.5% propyl nicotinate, less than about 0.1% propyl nicotinate, or less than about 0.01% propyl nicotinate. In a preferred embodiment, the composition comprises less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, less than about 0.1% nicotinic acid riboside, or less than about 0.01% nicotinic acid riboside. In some embodiments, the percentages are by weight.

[0136] In some embodiments, the pharma- ceutically acceptable excipient is selected from antiadherents, binders, coatings, dyes, disintegrants, flavorings, glidants, lubricants, preservatives, adsorbents, sweeteners, syrups, elixirs, dispersants, diluents, fillers, granulating agents, coatings, waxes, suspending agents, wetting agents, thickening agents, and vehicles, and combinations thereof. In some embodiments, the excipient is a solid excipient.

[0137] In some embodiments, the pharma- ceutically acceptable excipient is present in an amount of at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, or at least about 60% by weight of the composition. In some embodiments, the pharma- ceutically acceptable excipient is present in an amount of at least about 20%, at least about 25%, at least about 30%, at least about 35%, or at least about 40%, preferably at least about 30% by weight of the composition. In other embodiments, the pharma- ceutically acceptable excipient is present in an amount of at least about 50% by weight of the composition. The pH of the formulation can range from about 3 to about 11, but is usually from about 7 to about 10.

[0138] In some embodiments, the composition is in a solid form selected from tablets, pills, capsules, troches, granules, powders, sachets, dry powder inhalation forms, chewables, pastilles, and lozenges. In certain embodiments, the composition is in the form of a tablet. In other embodiments, the composition is in the form of a hard or soft gelatin capsule.

[0139] The compounds and crystalline solids of the present disclosure are formulated with conventional carriers and excipients that can be selected according to normal practice. Tablets can contain excipients, glidants, fillers, binders, etc. All formulations will optionally contain excipients such as those listed in the Handbook of Pharmaceutical Excipients (1986). Suitable excipients are also listed in the inactive ingredients database of the US Food and Drug Administration. Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkylcellulose, hydroxyalkylmethylcellulose, stearic acid, etc.

[0140] While it is possible for active pharmaceutical ingredients to be administered alone, it may be preferable to present them as pharmaceutical formulations. Both veterinary and human formulations of the present disclosure contain at least one active ingredient as defined above, together with one or more acceptable carriers therefor, and optionally other therapeutic ingredients. Some examples of materials that can function as pharma-ceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository wax; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil. , corn oil and soybean oil, (10) glycols, such as propylene glycol, (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol, (12) esters, such as ethyl oleate and ethyl laurate, (13) agar, (14) buffers, such as magnesium hydroxide and aluminum hydroxide, (15) alginic acid, (16) pyrogen-free water, (17) saline isotonic acid, (18) Ringer's solution, (19) ethyl alcohol, (20) phosphate buffers, and (21) other non-toxic compatible substances used in pharmaceutical formulations.

[0141] The pharmaceutical compositions (preparations) can be administered to a subject by any of several routes of administration, including, for example, oral (e.g., aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue), absorption through oral mucosa (e.g., sublingual), anal, rectal or vaginal (e.g., pessaries, creams or foams), parenteral (including, for example, intramuscular, intravenous, subcutaneous, or intrathecal as a sterile solution or suspension), intranasal, intraperitoneal, subcutaneous, subcutaneous, transdermal (e.g., a patch applied to the skin), and topical (e.g., as a cream, ointment or spray applied to the skin, or eye drops). The compound or crystalline solid may also be formulated for inhalation. In certain embodiments, the compound crystalline solid may simply be dissolved or suspended in sterile water. Details of suitable routes of administration and compositions suitable therefor can be found, for example, in U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, and patents cited therein.

[0142] Formulations of the present disclosure suitable for oral administration can be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient as a powder or granules. The active ingredient can also be administered as a bolus, electuary, or paste.

[0143] Tablets are made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active agent or dispersant. Molded tablets can be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. Tablets can be optionally coated or scored, and are optionally formulated to provide slow or controlled release of the active ingredient therefrom.

[0144] A pharmaceutical formulation according to the present disclosure includes a compound or crystalline solid according to the present disclosure together with one or more pharma- ceutically acceptable carriers or excipients, and optionally other therapeutic agents. A pharmaceutical formulation containing an active ingredient may be in any form suitable for the intended method of administration. When intended for oral use, for example, tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs may be prepared. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents, including sweeteners, flavoring agents, coloring agents, and preservatives, to provide a suitable preparation. Tablets containing the active ingredient in a mixture with non-toxic pharma- ceutically acceptable excipients suitable for the manufacture of tablets are acceptable. These excipients may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium or sodium phosphate, granulating and disintegrating agents such as corn starch or alginic acid, binders such as starch, gelatin or acacia, and lubricants such as magnesium stearate, stearic acid or talc. The tablets may be uncoated or may be coated by known techniques, such as microencapsulation, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained activity over a longer period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used alone or with a wax.

[0145] Formulations for oral use may also be presented as hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent, such as calcium phosphate or kaolin, or as soft gelatin capsules, in which the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0146] The aqueous suspensions of the present disclosure contain the active substance in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia, as well as dispersing or wetting agents such as naturally occurring phosphatides (e.g., lecithin), condensation products of alkylene oxides with fatty acids (e.g., polyoxyethylene stearates), condensation products of ethylene oxide with long-chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more sweetening agents such as sucrose or saccharin. Liquid formulations may also include eye drops or other forms of delivery to the ocular surface or adjacent locations such as the tear duct. Liquid formulations may include intravenous formulations, excipients, and carriers such as saline or buffer solutions, as well as packaging or containers for such formulations, injection or infusion, and the like.

[0147] Dispersible powders and granules of the present disclosure suitable for preparing aqueous suspension by adding water provide the active ingredient in a mixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives.Suitable dispersing or wetting agents and suspending agents are exemplified by those disclosed above.Additional excipients, such as sweeteners, flavoring agents, and coloring agents, can also be present.

[0148] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the subject being treated and the particular method of administration. For example, a time-release formulation intended for oral administration to humans may contain from about 1 to about 1000 mg of active agent compounded with an appropriate and convenient amount of carrier material, which may vary from about 5% to about 95% (weight:weight) of the total composition. Pharmaceutical compositions can be prepared to provide easily measurable amounts for administration.

[0149] Dosage will vary depending on the condition, age and weight of the patient, the nature and severity of the disorder to be treated or prevented, the route of administration, and the form of the drug, but generally, for an adult human patient, a daily dosage of 0.01 to 3000 mg of the compound or crystalline solid is recommended, which may be administered in a single dose or in divided doses. Generally, the compositions of the present disclosure may be provided in an aqueous solution containing, among other substances, about 0.1 to 30% w / v of the compound or crystalline solid disclosed herein for parenteral administration. Typical dosage ranges are about 0.01 to about 50 mg / kg of body weight per day, given in one or 2 to 4 divided doses. In certain embodiments, the compounds and / or crystalline solids described herein are administered in an amount of about 1 to about 3000 mg per day, about 100 to about 1000 mg per day, or about 250 to about 750 mg per day. If desired, the effective daily dose of the active compound or crystalline solid can be administered as 1, 2, 3, 4, 5, 6 or more separate doses administered at appropriate intervals throughout the day, optionally in unit dosage form. In some embodiments, the compounds and / or crystalline solids described herein are administered 1, 2, 3, 4, 5, 6 or more times per day. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be the amount of compound and / or crystalline solid that produces a therapeutic effect.

[0150] Formulations suitable for pulmonary or intranasal administration have particle sizes ranging from, for example, about 0.1 to about 500 microns, such as about 0.5, about 1, about 30, or about 35 microns, which are administered by rapid inhalation through the nasal passages or by inhalation through the mouth to reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder administration may be prepared according to conventional methods and may be delivered with other therapeutic agents.

[0151] The formulations may be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and may be stored in a freeze-dried (lyophilized) condition requiring only the addition of a sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit dosage formulations are those containing a daily dose or daily sub-dose, or an appropriate fraction thereof, of the active ingredient, as herein above recited.

[0152] It will be understood that in addition to the ingredients particularly mentioned above, the formulations of the present disclosure may include other agents conventional in the art having regard to the type of formulation in question, e.g., those suitable for oral administration may include flavoring agents.

[0153] In some embodiments, the amount of compound and / or crystalline solid in the composition is from about 0.001% to 100% by weight.

[0154] In some embodiments, the compound and / or crystalline solid is the only active pharmaceutical ingredient in the composition. Alternatively, the compound and / or crystalline solid is formulated in a composition with one or more additional active pharmaceutical ingredients. When formulated as the only active pharmaceutical ingredient, the compound and / or crystalline solid can be administered individually or as part of a regimen with one or more separately formulated active pharmaceutical ingredients.

[0155] When co-administered in the same formulation or as part of a regimen with one or more separately formulated active pharmaceutical ingredients, the additional active pharmaceutical ingredient may be selected from compounds of the NAD pathway, such as nicotinic acid (NA), nicotinamide (Nam), nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), nicotinic acid riboside (NAR), nicotinamide adenine dinucleotide (NAD / NADH), nicotinamide adenine dinucleotide phosphate (NADP), and nicotinic acid adenine dinucleotide (NaAD). In some embodiments, the compounds of formula I and II are administered together. In some embodiments, the additional active pharmaceutical ingredient is an amorphous solid. In some embodiments, the additional active pharmaceutical ingredient is a crystalline solid. In some embodiments, the additional active pharmaceutical ingredient is amorphous NMN. In some embodiments, the additional active pharmaceutical ingredient is crystalline NMN.

[0156] The invention having generally been described above will be more readily understood by reference to the following examples, which are included solely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention. EXAMPLES

[0157] Example 1.1: Preparation of Compound 2 [ka] A 50 mL collection flask was charged with 1.00 g (3.92 mmol) of nicotinic acid riboside and purged with argon. To this was added 6 mL of trimethyl phosphate, stirring was started, and the reaction was cooled in an ice bath. Then, 0.73 mL (7.84 mmol) of phosphorus oxychloride was added. After 30 min, the reaction was complete as determined by LC / MS. The reaction was added dropwise to 10 mL of ice-cold water. After the addition was complete, the mixture was concentrated in vacuo to remove approximately 9 g of solvent. The concentrated mixture was then added dropwise to a stirred ice-cold solution of 3.2 mL (23.0 mmol) of triethylamine in 50 mL of 1-propanol, resulting in a mixture with a pH = 3 as determined by pH paper. The suspension was stirred for 1 h, after which the solid precipitate was filtered and rinsed with 1-propanol to give the first crop of crude compound 2. After filtration was complete, a solid formed in the supernatant. The filtrate was stirred for 72 h, then the suspension was filtered and the precipitate was rinsed with 1-propanol to give a second crop of crude compound 2. The second crop was dried under vacuum and weighed 1.00 g while still wet. 1 H and 31 P NMR analysis showed that the second crop was more pure than the first crop. The second crop was used as seed material for subsequent experiments. See Figure 4. 1 H-NMR(500MHz;D2O):δ 9.42(s,1H),9.25(d,J=6.3Hz,1H),9.00(d,J=8.0Hz,1H),8.22(dd,J=7.8,6.5Hz,1H),6.16(d,J=5.3Hz,1H),4.58-4.56 (m,1H),4.49(t,J=5.1Hz,1H),4.37(dd,J=5.0,2.8Hz,1H),4.26-4.22(m,1H),4.12-4.08(d,d,d1H,J=12.0,5.1,2.2Hz)

[0158] Example 1.2A: Crystallization of Compound 2 To 100 mg of amorphous Compound 2 (prepared separately) was added 300 microliters of water. To this was added 350 microliters of 1-propanol, resulting in a cloudy solution. A second crop of Compound 2 from the experiment above was used to seed the mixture. After a few hours, crystals formed in the vial. The mixture was not filtered and was left as a slurry.

[0159] Example 1.2B: Crystallization of Compound 2 A 2.00 g quantity of amorphous Compound 2 (prepared separately) was dissolved in 6 mL of water, then 7 mL of 1-propanol was added to give a cloudy solution. An additional 2 mL of water was added to give a clear solution. A second crop of Compound 2 from the above experiment used to seed the mixture, but no crystals formed. An additional 0.2 mL of n-propanol was added to give a cloudy mixture. This was allowed to stir for a day, and no crystals were obtained. The mixture was then seeded with a drop of the slurry from the second experiment to prepare crystals of Compound 2, resulting in rapid crystallization. This was allowed to stir at ambient temperature for 2 days. The solid was filtered and washed with 15 mL (2:1 v:v) 1-propanol:water, 15 mL of 1-propanol, then 2×15 mL of methyl tert-butyl ether. The sample was dried under high vacuum at ambient temperature for 1 hour to give 1.92 g (96% mass recovery) of a white solid. The water solubility of the isolated product was approximately 20 mg / mL (50:1 w:w water:compound 2), in contrast to the starting compound 2 which was freely soluble in water (3:1 water:compound 2).

[0160] Example 1.2C: Crystallization of Compound 2 A 500 mg quantity of amorphous Compound 2 was dissolved in 1.5 mL of water. The sample completely dissolved and crystals began to form. The crystals were allowed to grow without stirring. The water was decanted and the solid was then dried under high vacuum. Single crystal X-ray diffraction quality crystals of Compound 2 were prepared. These crystals were used to generate an XRPD signature of Compound 2 crystals (Figure 2) and also to obtain a single crystal X-ray structure of Compound 2 (Figure 2C).

[0161] Example 2.1: Preparation of Compound 1 [ka] Compound 2 was filtered from 1-propanol prior to use and loaded into a 500 mL collection flask and purged with argon. The solid was suspended in 1-propanol and cooled in an ice bath. HCl gas was bubbled into the reaction mixture. The solid dissolved as the gas was bubbled into the suspension. The reaction was removed from the ice bath and stirred at room temperature for 3 days. When no starting material remained by LC / MS, the reaction mixture was concentrated on a rotary evaporator until an oil was obtained. An aliquot was placed under high vacuum for 30 minutes. The aliquot did not bubble. A sample of this was taken for LC / MS. The oil was diluted with 10 mL of 1-propanol and cooled in an ice bath. A total of 1.5 mL of triethylamine was added to bring the pH to 4-5. A significant amount of precipitate formed upon the addition of triethylamine. 40 mL of 1-propanol was added to dissolve all of the precipitate. Seed crystals of compound 1 were added to the solution. Over time, the solution became cloudy and a precipitate was observed to form. The mixture was stirred overnight at room temperature. After 4 hours on high VAC, an aliquot sample was taken for LC / MS. The flask contents appear more crystalline and filterable compared to the "milky" suspension. The solid was filtered. The filter cake was washed twice with 10 mL each of 1-propanol, followed by two 10 mL MTBE washes. A sample of the 1-propanol filtrate was taken for LC / MS. The solid was transferred to a vial. Wet weight was approximately 1.2 g. Placed under high vacuum for 1 hour. Obtained 1.23 g (55% yield) of white solid. A sample was analyzed by LC / MS, 1 H and 31 P NMR was performed, see Figure 3.

[0162] Example 2.2A: Polymorphism Screening Amorphous Compound 1 was screened for polymorphism according to the conditions provided in Table 1, including a 4-day maturation cycle between 0° C. (1 hour) and −20° C. (7 hours) between Observation 1 and Observation 2, and the results are shown.

[0163] [Table 1]

[0164] [Table 2]

[0165] Example 2.2B: Salt Screening Amorphous Compound 1 was screened for polymorphism according to Table 2. Compound 1 (15 mg) was weighed into an HPLC vial and a magnetic stir bar was added. The compound was dissolved in approximately 15 volumes (200 μl) EtOH at 5° C. with stirring at 500 rpm. Once dissolved, a molar equivalent of counterion was added and stirred at 5° C. (45 μl of 1 M stock or 90 μl of 0.5 M stock). The sample was cooled to −20° C. at 1° C. / min, but left in solution. The solution was allowed to slowly evaporate through a needle in the vial cap at 5° C.

[0166] [Table 3]

[0167] Example 2.2C: Co-crystal screening Amorphous Compound 1 was screened for polymorphism according to Table 3. Compound 1 (25 mg) was weighed into an HPLC vial and two milling balls were added. To this was added one molar equivalent of the conformer (as a solid). The mixture was first milled at 500 rpm and on a Fritsch planetary mill for 2 hours, then the collected solid was analyzed by XRPD. No crystallized material was obtained, the resulting solid was wetted with a drop of THF (7.5 μl) and milled on a Fritsch planetary mill for 2 hours at 500 rpm. Observations were made after milling and XRPD was performed on the collected solid.

[0168] [Table 4]

[0169] Example 2.2D: Salt Forms of Compound 1 Compound 1 (150 mg) was dissolved in 10 volumes (1.5 mL) of absolute EtOH with stirring at room temperature. After 3 min, a precipitate started to form. The sample was stirred for another 15 min before being filtered and dried under positive pressure. The sample was then placed in a vacuum oven at room temperature for 30 min under vacuum. The sample was then left in a fume hood overnight in a vial capped with perforated aluminum foil before being characterized. XPD pattern was obtained according to Figure 1A. See also Figure 1B. Percentage yield = approx. 63% and purity 98.7% by HPLC. Upon storage at 40°C / 75% RH for 7 days, the material was a sticky solid with a purity of 96.4% by HPLC. The sample is a fine white powder that is stable at room temperature, and although a small amount of solvent is trapped, analysis indicates that the sample is an anhydrous, non-solvated solid. The sample is stable under humidity conditions up to 70% RH.

[0170] FIG. 1C is a diagram of the crystal lattice unit cell of Compound 1, having the following attributes:

[0171] [Table 5]

[0172] Data for compound 1 were obtained by single crystal X-ray diffraction and the structure was solved by direct methods and refined using least-squares refinement. Atom assignments and positions in the crystal structure were assigned based on the electron density observed in Fourier difference maps and converged to a model that fit the experimental data. Non-hydrogen atoms were anisotropically refined, giving anisotropic displacement parameters (thermal ellipsoids) that can be seen in the ORTEP image (Figure 1C).

[0173] Example 2.2E: Scale-up of the Crystalline Form of Compound 1 Amorphous Compound 1 (1.23 g) was weighed into a 20 mL vial and treated with 7 volumes (8.60 mL) of methanol to obtain a clear solution. The solution was stirred at 400 rpm at 35° C. on a “polar bear” apparatus. The solution was supersaturated by adding 0.5 volumes (615 μl) of TBME and then seeded using pre-crystallized material (approximately 60 mg) that was retained in the solution. The sample was then cooled to 25° C. at 0.1° C. / min before the addition of the solvent. The addition of 2.6 volumes (3.2 mL) of TBME was performed over 50 min (1 μl / sec) using a syringe pump. The suspension was then cooled to 5° C. at 0.1° C. / min. The sample was kept at 5° C. for 1 h before being isolated using a Buchner funnel under vacuum. The sample was dried under vacuum for 20 min and then further dried in a vacuum oven overnight. The XPD pattern was obtained according to FIG. 1A. See also FIG. 1B. Percentage yield = approx. 45% and purity 97.9% by HPLC.

[0174] Example 3: Stability study of compound 2 Amorphous and crystalline forms of Compound 2 were prepared and stored under stress conditions to compare the stability of each form, as shown in Table 4. At T=0, amorphous Compound 2 assayed at 97.3% AUC and crystalline Compound 2 assayed at 99%. Samples were sealed in bottles containing saturated salt solutions to generate the required relative humidity, i.e., ammonium nitrate at RH=60%, sodium chloride at RH=75%, and saturated potassium nitrate at RH=97%. For RH=0%, solid phosphorus pentoxide was used. HPLC data was analyzed using a Waters Atlantis T3 C column equipped with an in-line guard column. 18 Collected using an Agilent 1290 system equipped with a column (3um, 100x4.6mm). Mobile phase A: 200mM ammonium carbonate (pH 3.8), mobile phase B: 95:5 MeOH:mobile phase A. Pumping is 1mL / min. Gradient is 0% B for 5 minutes followed by a 20 minute gradient to 100% B with a final hold of 3 minutes. Relevant data is collected via a DAD at 254nm.

[0175] [Table 6]

[0176] The results in Table 4 indicate that the crystalline form of Compound 2 has improved stability compared to the amorphous form of Compound 2.

[0177] INCORPORATION BY REFERENCE AND EQUIVALENTS All publications and patents mentioned herein are herein incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

[0178] While specific embodiments of the invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those of skill in the art upon review of this specification and the claims that follow. The full scope of the invention should be determined by reference to the claims, their full scope of equivalents, and the specification, together with such variations.

Claims

Crystalline solid comprising Compound 1 having 2θ values of 16.1, 20.1, and 24.

5. 【Chemical 1】

2. The crystalline solid according to claim 1, wherein the solid is anhydrous.

3. The crystalline solid according to claim 1, wherein the crystalline solid contains residual non-solvated solvent or residual non-hydrated water.

4. The crystalline solid according to claim 1, wherein the crystalline solid is non-hygroscopic.

5. A composition comprising the crystalline solid according to claim 1, the composition comprising less than about 5% by weight of propyl nicotinate.

6. A composition comprising the crystalline solid according to claim 1, the composition comprising less than about 5% by weight of Compound 2. 【Chemical Formula 2】

7. A composition comprising the crystalline solid according to claim 1, the composition comprising at least about 90% by weight of Compound 1.

8. A method for preparing the crystalline solid according to any one of claims 1 to 4, comprising: a) dissolving a solid comprising Compound 1 in a solvent to form a solution; and b) crystallizing Compound 1 from the solution to form the crystalline solid, wherein the solvent is methanol or ethanol.

9. The method according to claim 8, wherein the solvent is methanol.

10. The method according to claim 8, wherein the solution is anhydrous.

11. The method according to claim 8, wherein the solvent during the dissolving step is heated to about 30 to about 40 °C.

12. The method according to claim 8, wherein crystallization comprises forming a supersaturated solution from the solution, the supersaturated solution being supersaturated with respect to Compound 1.

13. The method according to claim 12, wherein forming the supersaturated solution comprises adding an anti-solvent to the solution, reducing the temperature of the solution, reducing the volume of the solution, or any combination thereof.

14. The method according to claim 8, wherein crystallization comprises adding a seed crystal to the solution, the seed crystal comprising Compound 1.

15. Compound 1: [Chemical Formula 3] A method for purifying, the method comprising: a) dissolving a solid comprising Compound 1 in a solvent to form a solution; and b) crystallizing Compound 1 from the solution to form a solid having a higher purity than the solid dissolved in step (a), wherein the solid having the higher purity is a crystalline solid of Compound 1, and the solvent is methanol or ethanol. **Claim 16**: The method according to claim 15, wherein step (a) is carried out using the amorphous compound 1, and the solid of higher purity in step (b) has a purity of more than about 95%. **Claim 17**: A crystalline solid comprising a compound 2 having 2θ values of 21.5, 24.2, 26.7, and 19.

6. 【Chemical 3】 **Claim 18** **Claim 19**: The crystalline solid according to claim 17, wherein the compound 2 is a hydrate. **Claim 20** **Claim 21**: The crystalline solid according to claim 17, wherein the crystalline solid contains residual non-solvated solvent or residual non-hydrated water. **Claim 22** **Claim 23**: A composition comprising the crystalline solid according to claim 17 and containing less than about 5% by weight of nicotinic acid. **Claim 24** **Claim 25**: A composition comprising the crystalline solid according to claim 17 and containing less than about 1% of nicotinic acid riboside. **Claim 26** **Claim 27**: A composition comprising the crystalline solid according to claim 17 and containing at least about 90% of the compound 2. **Claim 28** **Claim 29**: A method for preparing the crystalline solid according to any one of claims 17 to 19, comprising: a) dissolving the compound 2 in a solvent to form a solution; b) crystallizing the compound 2 from the solution to form the crystalline solid, wherein the solvent contains water. **Claim 30**: The method according to claim 23, wherein the solvent further contains alcohol. **Claim 31** **Claim 32**: The method according to claim 30, wherein the alcohol is 1-propanol. **Claim 33** **Claim 34**: The method according to claim 23, wherein the temperature of the solvent during the dissolving step is ambient temperature. **Claim 35** **Claim 36**: The method according to claim 23, wherein crystallization includes forming a supersaturated solution from the solution, and the supersaturated solution is supersaturated with respect to the compound 2. **Claim 37** **Claim 38**: The method according to claim 36, wherein forming the supersaturated solution includes adding an antisolvent to the solution, reducing the temperature of the solution, reducing the volume of the solution, or any combination thereof. **Claim 39** **Claim 40**: The method according to claim 23, wherein crystallization includes adding a seed crystal to the solution, and the seed crystal contains the compound 2. **Claim 41** **Claim 42**: A method for purifying the compound 2, the method comprising: a) dissolving the compound 2 in a solvent to form a solution; **Claim 43** b) crystallizing the compound 2 from the solution to form a solid of higher purity than the compound dissolved in step (a), wherein the solvent contains water. 【Chemical 4】 **Claim 44** **Claim 45**: The method according to claim 42, wherein the solvent further contains alcohol. **Claim 46** **Claim 47**: The method according to claim 45, wherein the alcohol is 1-propanol. **Claim 48** The method according to claim 30, wherein step (a) is carried out using an amorphous solid and the higher purity solid in step (b) is of a purity of greater than about 95% with respect to said Compound 2.

32. A pharmaceutical composition comprising the crystalline solid according to claim 1 or 17 and one or more pharmaceutically acceptable excipients.

33. Use of the crystalline solid according to claim 1 or 17, or a composition according to any one of claims 5 to 7 and 20 to 22, in the manufacture of a medicament for increasing NAD levels in a subject.

34. Use of the crystalline solid according to claim 1 or 17, or a composition according to any one of claims 5 to 7 and 20 to 22, in the manufacture of a medicament for modulating NAD levels in a subject.

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