Method for Producing Crystalline Beta Nicotinamide Riboside Triacetate Chloride

Abstract

The present disclosure relates to a process for producing crystalline beta-nicotinamide riboside triacetate chloride having improved physical characteristics. A substantially crystalline beta-nicotinamide riboside triacetate chloride or a salt or solvate thereof is described having a chemical purity of greater than about 90% (w / w) and containing less than about 5000 ppm ethanol.

Description

[Technical field]

[0001] Technical Field The present invention relates to a process for producing crystalline beta-nicotinamide riboside triacetate chloride having improved physical characteristics. [Background technology]

[0002] background Nicotinamide riboside (NR) is a valuable biologically active intermediate. This compound is involved in the synthesis of NAD + It has been shown to be involved in processing and metabolic pathways involving .

[0003] Dietary vitamin B3, which includes nicotinamide ("Nam" or "NM"), nicotinic acid ("NA"), and nicotinamide riboside ("NR"), acts as a coenzyme to regulate the synthesis of nicotinamide adenine dinucleotide ("NAD - " ), its phosphorylated parent (" NADP + " or "NAD(P) + ") and its respective reduced forms (NADH and NADPH, respectively). In cells, NAD(P) + Once converted to NAD(P)H, vitamin B3 metabolites control metabolism as they are used as cosubstrates in the modification processes of numerous intracellular proteins that control numerous essential signaling events (e.g., ribosylation and deacetylation of adenosine diphosphate) and as cofactors in over 400 oxidoreductase reactions. This is demonstrated by a wide range of metabolic endpoints, including deacylation of key regulatory metabolic enzymes, leading to restoration of mitochondrial activity and oxygen consumption. Critically, mitochondrial dysfunction and cellular damage have been correlated with depletion of the NAD(P)(H)-cofactor pool when it is present at suboptimal intracellular concentrations. Vitamin B3 deficiency reduces NAD(P)(H)-cofactor levels, which is a key factor in the regulation of mitochondrial function. +(P) depletion results in impaired cellular activity, and supplementation with NA, Nam, NR, and nicotinamide mononucleotide ("NMN") provides additional NAD(P) + The beneficial effects of bioavailability are primarily observed in cells and tissues with compromised metabolism and mitochondrial function.

[0004] Despite extensive optimization of solution-based methodologies for the preparation of nucleotides over the years, the synthesis of nicotinoyl riboside, monophosphorylation of its active hydroxyl group, and its subsequent conjugation remain challenging and problematic in terms of low yields, product stability, and isolation from polar solvents.Current methodologies also suffer from atom and energy inefficiencies, for example due to the use of large solvent excesses and the need for temperature-controlled reaction conditions.

[0005] Reported syntheses of nicotinamide riboside (NR) have become feasible on a larger scale, but still exhibit batch-to-batch quality variability and difficulty in maintaining good specifications, due to the use of corrosive and expensive reagents and lengthy deprotection steps.

[0006] Partially protected nucleosides and nucleotides have found a wide range of applications to achieve improved bioavailability of nucleoside and nucleotide parents. Such partial protection includes the introduction of hydrolyzable phosphoramidates or the modification of mixed anhydrides of phosphoric acid monoesters in the form of Protide and CycloSal derivatives, as well as the modification of hydroxyl groups with esters, carboxylates, and acetyl groups. While the previous types of protection are becoming more feasible on a larger scale, modifications at the phosphorus center remain difficult to achieve on a larger scale, especially for nucleoside entities that are highly sensitive to changes in pH and easily decompose by heat.

[0007] Reduced nicotinamide riboside ("NRH") is the cytosolic NAD +It has been consistently shown to be more effective at increasing NAD levels, surpassing nicotinamide riboside (NR) in this respect. While their physiological and potentially therapeutic roles have yet to be explored due to a lack of accessible material in sufficient quantities for extensive study, phosphorylated forms of NRH and reduced nicotinic acid riboside ("NARH") or their derivatives have been shown to be more effective at increasing NAD levels. + It is expected that boost capabilities will be similar.

[0008] Reported syntheses of reduced nicotinamide riboside (NRH) are becoming more widely available, but remain performed on a small scale using corrosive and expensive reagents and lengthy deprotection steps, and therefore still exhibit batch-to-batch quality variability and difficulty in maintaining good specifications. In the current discussion, reduced nicotinamide riboside (NRH) generally refers to the "reduced pyridine" nucleus, and more specifically, to 1,4-dihydropyridine compounds.

[0009] Synthetically, the preparation of 5'-nucleotides remains time-consuming, atom-inefficient, and costly due to the need for numerous protection and deprotection steps. In these preparation methods, the required chlorodialkylphosphate, tetraalkylpyrophosphate, chlorophosphite, or phosphoramidite reagents are also expensive starting materials due to their chemical functionalization and chemical instability, and thus the resulting associated synthetic difficulties. Phosphorylation reaction conditions are difficult to control and often use unapproved or toxic organic solvents, limiting the market for the compounds produced.

[0010] One known alternative approach to the protection / deprotection method is to use phosphorus oxychloride (P(O)Cl3) (i.e., Yoshikawa conditions). However, there are still drawbacks to this method, as follows: Without being bound by theory, in this method, a polar trialkyl phosphate solvent, such as P(O)(OMe)3, is used in large excess, which is believed to enhance the reaction rate while limiting the undesired reactivity of P(O)Cl3 as a chlorinating agent. Hence, the use of excess P(O)Cl3 / P(O)(OR)3 is believed to be a better combination for chemoselective 5'-O-phosphorylation of unprotected ribosides. However, the use of trialkyl phosphate solvents, such as P(O)(OMe)3, prevents implementation for the preparation of materials for final human use, as this solvent class is highly toxic (known carcinogen, approved non-GRAS) and difficult to remove from the polar end product. See M. Yoshikawa et al., Studies of Phosphorylation. III, Selective Phosphorylation of Unprotected Nucleosides, 42 BULL. CHEM. SOC. JAPAN 3505 (1969); Jaemoon Lee et al., A chemical synthesis of nicotinamide adenine dinucleotide (NAD+), CHEM. COMMUN. 729 (1999), each of which is incorporated herein by reference in its entirety.

[0011] Nicotinamide adenine dinucleotide (NAD + ) remains an expensive cofactor and its commercial availability is limited simply by its complex chemistry and the highly reactive pyrophosphate bond, which makes it difficult to make on a large scale.

[0012] Nicotinoyl riboside, such as nicotinamide riboside (NR) and nicotinic acid riboside ("NAR"), nicotinamide mononucleotide (NMN), and NAD+ are seen as useful bioavailable precursors of the NAD(P)(H) pool to combat and treat a wide range of non-communicable diseases, especially those associated with mitochondrial dysfunction and impaired cellular metabolism. Therefore, optimizing the large-scale synthesis of these vitamin B3 derivatives would be highly valuable to make these compounds more widely available to society, both as nutritional and pharmaceutical entities.

[0013] Reduced nicotinoyl ribosides, such as reduced nicotinamide riboside (NRH), reduced nicotinic acid riboside (NARH), reduced nicotinamide mononucleotide ("NMNH"), reduced nicotinic acid mononucleotide ("NaMNH"), and reduced nicotinamide adenine dinucleotide ("NADH"), are seen as useful bioavailable precursors of the NAD(P)(H) pool to combat and treat a wide range of non-communicable diseases, particularly those associated with mitochondrial dysfunction and impaired cellular metabolism. Therefore, optimizing the large-scale synthesis of these vitamin B3 derivatives is highly valuable in making these compounds more widely available to society, both as nutritional and pharmaceutical entities.

[0014] Crystalline forms of useful molecules may have advantageous properties compared to the respective amorphous forms of such molecules. For example, crystalline forms are often easier to handle and process, for example, when preparing compositions that include the crystalline form. Crystalline forms typically have greater storage stability and are more amenable to purification. The use of crystalline forms of pharma-ceutically useful compounds can also improve the performance characteristics of pharmaceutical products that include the compound. Obtaining crystalline forms can also help formulation scientists to expand the repertoire of materials available for formulation optimization, for example, by providing products with different properties, such as better processing or handling characteristics, improved dissolution profiles, or improved shelf life.

[0015] WO 2016 / 014927 A2, which is incorporated herein by reference in its entirety, describes crystalline forms of nicotinamide riboside, including nicotinamide riboside chloride Form I. Also disclosed are pharmaceutical compositions comprising crystalline form I of nicotinamide riboside chloride, and methods of making such pharmaceutical compositions.

[0016] WO 2016 / 144660A1, which is incorporated herein by reference in its entirety, describes crystalline forms of nicotinamide riboside, including nicotinamide riboside chloride Form II. Also disclosed are pharmaceutical compositions comprising crystalline nicotinamide riboside chloride Form II, and methods of producing such pharmaceutical compositions.

[0017] In view of the foregoing, there is a need for an atom-efficient process in terms of reagents and solvent equivalents that avoids the need for polar, non-GRAS ("Generally Recognized As Safe") solvents, is versatile with respect to limitations associated with solubility and reagent mixing, is time-efficient, and is energy-efficient, and provides an efficient, practical, and scalably feasible method for the preparation of nicotinoyl riboside, reduced nicotinoyl riboside, its modified derivatives, its phosphorylated analogues, and its adenylyl dinucleotide conjugates.

[0018] In view of the foregoing, there is a need for novel crystalline forms of nicotinoyl riboside, reduced nicotinoyl riboside, its modified derivatives, its phosphorylated analogues, and its adenylyl dinucleotide conjugates.

[0019] Nicotinic acid and nicotinamide (collectively niacin) are the building blocks of nicotinamide adenine dinucleotide (NAD + ) is a vitamin form of tryptophan. Eukaryotes convert tryptophan to NAD via the kynureline pathway. +(Krehl, et al. Science (1945) 101:489-490; Schutz and Feigelson, J. Biol. Chem. (1972) 247:5327-5332), and niacin supplementation prevents pellagra, which can occur in populations with a tryptophan-deficient diet. Thus, nicotinic acid is phosphoribosylated to nicotinic acid mononucleotide (NaMN), which is subsequently adenylylated to form nicotinic acid adenine dinucleotide (NaAD), which is then amidated to NAD + It is well established that endothelial cells are formed (Preiss and Handler (1958) 233:488-492; Ibid., 493-50).

[0020] Nicotinamide adenine dinucleotide (NAD + ") is an essential enzyme cofactor for the function of several enzymes involved in reduction-oxidation reactions and energy metabolism (Katrina L. Bogan & Charles Brenner, Nicotinic Acid, Nicotinamide, and Nicotinamide Riboside: A Molecular Evaluation of NAD+ Precursor Vitamins in Human Nutritions, 28 Annual Review of Nutrition 115 (2008)). NAD + NAD functions as an electron carrier in the cellular metabolism of amino acids, fatty acids, and carbohydrates (Bogan & Brenner 2008). +NAD acts as an activator and substrate for sirtuins, a family of protein deacetylases that have been implicated in metabolic function and longevity in lower organisms (Laurent Mouchiroud et al., The NAD+ / Sirtuin Pathway Modulates Longevity through Activation of Mitochondrial UPR and FOXO Signaling, 154 Cell 430 (2013)). + The coenzyme activity of β-glucose, which is subject to tight regulation of its biosynthesis and bioavailability, represents an important metabolic surveillance system that is clearly involved in the aging process.

[0021] In cells, NAD(P) + Once converted to , vitamin B3 is used as a cosubstrate in two intracellular modifications that control numerous essential signaling events (adenosine diphosphate ribosylation and deacetylation) and is a cofactor for over 400 reduction-oxidation enzymes, thus controlling metabolism. This is demonstrated by a wide range of metabolic endpoints, including deacetylation of key regulatory proteins, increased mitochondrial activity, and oxygen consumption. Critically, the NAD(P)(H)-cofactor family can promote mitochondrial dysfunction and cellular damage if present at suboptimal intracellular concentrations. Vitamin B3 deficiency increases NAD + This results in impaired cellular activity manifested through depletion and the need for additional NAD by supplementation with nicotinic acid ("NA"), nicotinamide ("Nam"), and nicotinamide riboside ("NR"). + The beneficial effects of bioavailability are primarily observed in cells and tissues with compromised metabolism and mitochondrial function.

[0022] Interestingly, supplementation with nicotinic acid ("NA") and nicotinamide ("Nam") is important in acute vitamin B3 deficiency, and at the cellular level, all three metabolites are NAD +Although biosynthetic, it does not demonstrate the same physiological outcomes compared to nicotinamide riboside ("NR") supplementation, highlighting the complexity of the pharmacokinetics and biodistribution of B3-vitamin components.

[0023] Intracellular NAD + The bulk of the circulating supply of NAD is thought to be regenerated through the efficient salvage of nicotinamide ("Nam"), while de novo NAD + is derived from tryptophan (Anthony Rongvaux et al., Reconstructing eukaryotic NAD metabolism, 25 BioEssays 683 (2003)). Crucially, these salvage and de novo pathways apparently catalyze the conversion of NAD via phosphoriboside pyrophosphate intermediates, depending on the functional form of vitamins B1, B2, and B6. + Nicotinamide riboside ("NR") produces NAD + is the only form of vitamin B3 that can be produced independently from vitamins B1, B2, and B6, and is the only form of NAD + The salvage pathway that uses nicotinamide riboside ("NR") for the production of is expressed in most eukaryotes.

[0024] The main NAD that feeds the salvage route + The precursors are nicotinamide ("Nam") and nicotinamide riboside ("NR") (Bogan & Brenner 2008). Research has shown that nicotinamide riboside ("NR") promotes the synthesis of NAD through the formation of nicotinamide mononucleotide ("NMN"). + Shortly after entry into the cell, nicotinamide riboside ("NR") is phosphorylated by NR kinase ("NRK") to generate NMN, which is subsequently converted to NAD by nicotinamide mononucleotide adenylyltransferase ("NMNAT"). + (Bogan & Brenner 2008). NMN is converted to NAD in mitochondria.+ Nicotinamide ("Nam") and nicotinamide riboside ("NR") are the only metabolites that can be converted to NAD + Two candidate NADs that can improve mitochondrial fuel oxidation by supplementing + The key difference is that nicotinamide riboside ("NR") is a precursor of NAD, bypassing the rate-limiting step in the salvage pathway, nicotinamide phosphoribosyltransferase ("NAMPT"). + Nicotinamide ("Nam") is a direct two-step pathway for the synthesis of NAD. + This is because nicotinamide riboside ("NR") is a highly effective NAD + This reinforces the fact that dietary NAD is a precursor. + Deficiency of the precursor and / or tryptophan causes pellagra, a disease characterized by dermatitis, diarrhea, and dementia (Bogan & Brenner 2008). In summary, NAD + NAD is required for normal mitochondrial function. Mitochondria are the powerhouses of the cell, so NAD + is required for energy production within cells.

[0025] NAD + NAD was first characterized as a coenzyme for oxidoreductase enzymes. + Although the conversion between NADH, NADP, and NADPH does not involve the loss of the entire coenzyme, + It has also been discovered that sirtuin enzymes, such as Sir2 in S. cerevisiae, and their homologs, act to regulate NAD +Sir2 deacetylates lysine residues at the expense of NAD equivalents, and this activity is required for Sir2 function as a transcriptional silencer (Imai, et al., Cold Spring Harb. Symp. Quant. Biol. (2000) 65:297-302). + NAD-dependent deacetylation is required not only for altering gene expression, but also for suppression of ribosomal DNA recombination and for extending life span in response to calorie restriction (Lin, et al., Science (2000) 289:2126-2128; Lin, et al., Nature (2002) 418:344-348). + is consumed by Sir2 to generate a mixture of 2'- and 3'O-acetylated ADP-ribose plus nicotinamide, and deacetylated polypeptides (Sauve, et al., Biochemistry (2001) 40:15456-15463). Additional enzymes, including poly(ADP-ribose) polymerase and cADP-ribose synthase, also mediate the uptake of NAD. + dependent and produce nicotinamide and ADP-ribosyl products (Ziegler, Eur. J. Biochem. (2000) 267:1550-1564; Burkle, Bioessays (2001) 23:795-806).

[0026] US Patent No. 9,975,915 (incorporated herein by reference in its entirety) describes crystalline forms of nicotinamide riboside, including NR methanolate form II of nicotinamide riboside chloride. Also disclosed are compositions comprising NR methanolate form II of nicotinamide riboside chloride, and methods for preparing NR methanolate form II of nicotinamide riboside chloride. Also disclosed are crystalline forms of nicotinic acid riboside (NAR), including nicotinic acid riboside (NAR) form I. Also disclosed are compositions comprising nicotinic acid riboside (NAR) form I, and methods for preparing nicotinic acid riboside (NAR) form I. Also disclosed are crystalline forms of nicotinamide riboside triacetate (1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-nicotinamide, "NR triacetate," or "NRTA" (also known as "NRT")), including Form I of nicotinamide riboside triacetate (NRTA) chloride ("NRTA-Cl"). Also disclosed are compositions comprising Form I of nicotinamide riboside triacetate (NRTA), and methods of preparing Form I of nicotinamide riboside triacetate (NRTA). Also disclosed are crystalline forms of nicotinic acid riboside triacetate (1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-nicotinic acid, "NAR triacetate," or "NARTA"), including Form I of nicotinamide riboside triacetate (NARTA). Also disclosed are compositions comprising nicotinic acid riboside triacetate (NARTA) Form I, and methods for preparing nicotinic acid riboside triacetate (NARTA) Form I. Also disclosed are crystalline forms of nicotinamide mononucleotide ("NMN"), including nicotinamide mononucleotide (NMN) Form III and nicotinamide mononucleotide (NMN) Form IV.Also disclosed are compositions comprising nicotinamide mononucleotide (NMN) form III and compositions comprising nicotinamide mononucleotide (NMN) form IV, as well as methods for preparing nicotinamide mononucleotide (NMN) form III and methods for preparing nicotinamide mononucleotide (NMN) form IV.

[0027] Nicotinamide riboside chloride is known to exist as two stable polymorphs, Form I and Form II. Known synthesis and purification procedures have been shown to produce mixtures of Form I and Form II with poor physical properties that present difficulties in downstream encapsulation processing.

[0028] In addition, nicotinamide riboside chloride triacetate chloride (NRTA-Cl) is known to exist as a stable polymorph, namely Form I. There are known challenges in the large-scale production of NRTA-Cl, including identifying a scalable crystallization process.

[0029] The present invention seeks to solve these problems and others. Summary of the Invention [Means for solving the problem]

[0030] The invention referred to provides process conditions that have been shown to improve particle size distribution, bulk density, and polymorph control for the production of nicotinamide riboside triacetate chloride.

[0031] In one embodiment, a substantially crystalline nicotinamide riboside triacetate compound, or a salt or solvate thereof, is described having a chemical purity of greater than about 90% (w / w) and containing less than about 5000 ppm ethanol. In a further embodiment, the substantially crystalline nicotinamide riboside triacetate compound is substantially the beta anomeric form of nicotinamide riboside triacetate chloride.

[0032] In another embodiment, a method for producing a nicotinamide riboside triacetate compound, or a salt or solvate thereof, is described, comprising: (a) adding a quantity of crude nicotinamide riboside triacetate to a volume of a first solvent to form a reaction mixture; (b) heating the reaction mixture to a temperature of about 20° C. to about 60° C.; (c) cooling the reaction mixture; (d) adding a second solvent; and (e) isolating the substantially crystalline compound nicotinamide riboside triacetate or a salt or solvate thereof as a crystalline powder. Optionally, the method may include a step (c1) of seeding the reaction mixture with the crystalline compound nicotinamide riboside triacetate or a salt or solvate thereof after step (c). [Brief description of the drawings]

[0033] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] FIG. 1 is a schematic diagram of the synthetic sequence used to produce crystalline beta-nicotinamide riboside triacetate chloride. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0034] Nicotinamide riboside ("NR") has the formula (I): [ka] It is a pyridinium compound having the formula:

[0035] NR in formula (I) may include salts or solvates. Salts include a counter ion ("X") selected from chloride, bromide, iodide, etc. -"). For example, a useful monosalt is the chloride salt of NR ("NR-Cl"). Additional salts include fluoride, formate, acetate, propionate, butyrate, glutamate, aspartate, ascorbate, benzoate, carbonate, citrate, carbamate, gluconate, lactate, methyl bromide, methyl sulfate, nitrate, phosphate, diphosphate, succinate, sulfate, tartrate, bitartrate, malate, bitamate, maleate, fumarate, stearate, palmitate, myristate, laurate, caprate, caprylate, caproate, oleate, linoleate, sulfonate, trifluoromethanesulfonate, trichloromethanesulfonate, tribromo ... Examples of suitable hydroxyl groups include, but are not limited to, sulfonates, trichloroacetates, tribromoacetates, trifluoroacetates, glycolates, glucuronates, pyruvates, anthranilates, 4-hydroxybenzoates, phenylacetates, mandelates, pamoates, methanesulfonates, ethanesulfonates, benzenesulfonates, pantothenates, 2-hydroxyethanesulfonates, p-toluenesulfonates, sulfanilates, cyclohexylaminesulfonates, alginates, beta-hydroxybutyrates, salicylates, galactarates, galacturonates, and the like. - In some cases, such as for NAR, NAMN, and NMN, when is absent, the counterion is an inner salt.

[0036] NR is hydrophilic but susceptible to hydrolysis. This presents a unique requirement for chemical stability necessitating the microencapsulation of water-soluble compounds. This is a reversal of the common technique of formulators to microencapsulate hydrophobic, lipophilic, or water-insoluble materials to provide better bioavailability.

[0037] In a further embodiment, a derivative of NR having formula (Ia) or a salt, solvate, or prodrug thereof is contemplated: [ka] In the formula, R 6 is hydrogen, -C(O)R', -C(O)OR', -C(O)NHR', substituted or unsubstituted (C1-C 24 ) alkyl, substituted or unsubstituted (C3-C8) cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, and substituted or unsubstituted heterocycle; R' is hydrogen, -(C1-C 24 ) alkyl, -(C3-C8) cycloalkyl, aryl, heteroaryl, heterocyclic, aryl (C1-C 24 ) alkyl, and heterocyclic (C1-C 24 ) alkyl; R 7 and R 8 are independently hydrogen, -C(O)R', -C(O)OR', -C(O)NHR', substituted or unsubstituted (C1-C 24 )alkyl, substituted or unsubstituted (C3-C8)cycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heterocycle, substituted or unsubstituted aryl(C1-C4)alkyl, and substituted or unsubstituted heterocycle(C1-C4)alkyl.

[0038] The present disclosure also relates to other NADs. + Precursors include, but are not limited to, one or more nicotinyl riboside compounds selected from nicotinic acid riboside (NAR, II), nicotinamide mononucleotide (NMN, III), nicotinic acid mononucleotide (NaMN, IV), reduced nicotinamide riboside (NRH, V), reduced nicotinic acid riboside (NARH, VI), NR triacetate (NRTA, VII, which is a type of Ia), NAR triacetate (NARTA, VIII), NRH triacetate (NRH-TA, IX), or NARH triacetate (NARH-TA, X), and salts, solvates, or mixtures thereof, or derivatives thereof.

[0039] Nicotinic acid riboside (NAR) has the formula (II): [ka] is a pyridinium nicotinyl compound having the formula In some cases, X - is absent and NAR is an inner salt (zwitterionic species).

[0040] Nicotinamide mononucleotide (NMN) has the formula (III): [ka] is a pyridinium nicotinyl compound having the formula In some cases, X - is absent and NMN can be an inner salt.

[0041] Nicotinic acid mononucleotide (NaMN) has the formula (IV): [ka] is a pyridinium nicotinyl compound having the formula In some cases, X - is absent and NaMN can be an inner salt.

[0042] The salts may include chloride, bromide, iodide, etc., or other counterions selected from the organic counterions shown in formula (I) ("X - "). For example, a useful monosalt is the chloride salt of NR ("NR-Cl"). Additional salts, including phosphates, can include, but are not limited to, one or more of sodium, potassium, lithium, magnesium, calcium, strontium, or barium. Reduced nicotinamide riboside ("NRH") has the formula (V): [ka] is a 1,4-dihydropyridyl reduced nicotinyl compound having the formula:

[0043] Reduced nicotinic acid riboside ("NARH") has the formula (VI): [ka] is a 1,4-dihydropyridyl reduced nicotinyl compound having the formula:

[0044] In certain species of compound (Ia), the free hydrogen of the hydroxyl group on the ribose moiety of nicotinamide riboside (NR, I) is replaced with an acetyl group (CH3-C(=O)-) to give the compound of formula (VII): [ka] and forming 1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-nicotinamide ("NR triacetate" or "NRTA") having the formula: X - is defined above.

[0045] The free hydrogen of the hydroxyl group on the ribose moiety of nicotinic acid riboside (NAR, II) is replaced with an acetyl group (CH3-C(=O)-) to give the compound of formula (VIII): [ka] and forming 1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-nicotinic acid ("NAR triacetate" or "NARTA") having the formula: In some cases, X - is absent and NARTA is an internal salt.

[0046] The free hydrogen of the hydroxyl group on the ribose moiety of reduced nicotinamide riboside (NRH, V) is replaced with an acetyl group (CH3-C(=O)-) to give the compound of formula (IX): [ka] In one embodiment, 1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-1,4-dihydronicotinamide ("NRH triacetate" or "NRH-TA") can be formed having the formula:

[0047] The free hydrogen of the hydroxyl group on the ribose moiety of reduced nicotinic acid riboside (NARH, VI) is replaced with an acetyl group (CH3-C(=O)-) to form the compound of formula (X): [ka] In one embodiment, 1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-1,4-dihydronicotinic acid ("NARH triacetate" or "NARH-TA") can be formed having the formula:

[0048] nicotinamide riboside (NR, I), nicotinic acid riboside (NAR, II), nicotinamide mononucleotide (NMN, III), nicotinic acid mononucleotide (NaMN, IV), reduced nicotinamide riboside (NRH, V), reduced nicotinic acid riboside (NARH, VI), nicotinamide riboside triacetate (NRTA, VII), nicotinic acid riboside triacetate (NARTA, VIII), reduced nicotinamide riboside triacetate (NRH-TA, IX), and reduced nicotinic acid riboside triacetate (NARH-TA, X), optionally with X as a counter ion. - is absent or X - If present, X -are fluoride ion, formate ion, acetate ion, propionate ion, butyrate ion, glutamate ion, aspartate ion, ascorbate ion, benzoate ion, carbonate ion, citrate ion, carbamate ion, gluconate ion, lactate ion, methyl bromide, methyl sulfate, nitrate ion, phosphate ion, diphosphate ion, succinate ion, sulfate ion, tartrate ion, hydrogen tartrate ion, malate ion, hydrogen malate ion, maleate ion, fumarate ion, citrate ion, stearate ion, palmitate ion, myristic acid ion, laurate ion, caprate ion, caprylate ion, caproate ion, oleate ion, linoleic acid ion, sulfonate ion, trifluoromethanesulfonic acid ions, trichloromethanesulfonate ions, tribromomethanesulfonate ions, trichloroacetate ions, tribromoacetate ions, trifluoroacetate ions, glycolate ions, glucuronate ions, pyruvate ions, anthranilate ions, 4-hydroxybenzoate ions, phenylacetate ions, mandelate ions, pamoate ions, methanesulfonate ions, ethanesulfonate ions, benzenesulfonate ions, pantothenate ions, 2-hydroxyethanesulfonate ions, p-toluenesulfonate ions, sulfanilate ions, cyclohexylaminesulfonate ions, alginate ions, beta-hydroxybutyrate ions, salicylate ions, galactarate ions, galacturonate ions, and the like; In some cases, X - is absent, in some cases the counterion is an internal salt; In some cases, X - is the anion of a substituted or unsubstituted carboxylic acid selected from monocarboxylic acids, dicarboxylic acids, or polycarboxylic acids; In some cases, X -is an anion of a substituted monocarboxylic acid, and also optionally an anion of a substituted propanoic acid (propanoate anion or propionate anion), or an anion of a substituted acetic acid (acetate anion), or an anion of a hydroxyl group propanoic acid, or an anion of 2-hydroxypropionic acid (which is lactic acid; the anion of lactic acid is the lactate anion), or a trihaloacetate selected from trichloroacetate, tribromoacetate, or trifluoroacetate; In some cases, X - is an anion of an unsubstituted monocarboxylic acid selected from formic acid, acetic acid, propionic acid, or butyric acid, or an anion of an unsubstituted monocarboxylic acid having a carbon chain length of C6 to C 24 and anions of long chain fatty acids including saturated, unsaturated, and polyunsaturated fatty acids (e.g., stearic acid, palmitic acid, myristic acid, lauric acid, capric acid, caprylic acid, caproic acid, oleic acid, linoleic acid, omega-6 fatty acids, omega-3 fatty acids) (the anions being formate, acetate, propionate, butyrate, and stearate, respectively); In some cases, X - is optionally selected from glutamic acid and aspartic acid (anions which are glutamate and aspartate, respectively), or alternatively is the anion of a substituted or unsubstituted amino acid, i.e., an amino-monocarboxylic acid or an amino-dicarboxylic acid, selected from alanine, beta-alanine, arginine, asparagine, cysteine, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, or tyrosine; In some cases, X - is the anion of ascorbic acid (ascorbate anion); In some cases, X - is a halide selected from fluoride, chloride, bromide, or iodide; In some cases, X -is the anion of a substituted or unsubstituted sulfonate, and optionally a trihalomethanesulfonate selected from trifluoromethanesulfonate, tribromomethanesulfonate, or trichloromethanesulfonate; In some cases, X - is the anion of a substituted or unsubstituted carbonate, and optionally also of hydrogen carbonate.

[0049] In yet another embodiment, the present disclosure relates to crystalline forms of nicotinic acid riboside (NAR), including but not limited to "Form II" or "Form I" of nicotinic acid riboside (1-(beta-D-ribofuranosyl)-nicotinic acid, NAR), and methods for their preparation, as disclosed in U.S. Pat. No. 11,214,589 and U.S. Pat. No. 9,975,915, respectively.

[0050] In yet another embodiment, the present disclosure relates to a crystalline form of nicotinamide riboside triacetate chloride (NRTA-Cl) Form I, and methods of its preparation, as disclosed in U.S. Pat. No. 9,975,915.

[0051] In yet another embodiment, the present disclosure relates to crystalline forms of nicotinic acid riboside triacetate ("NARTA"), including but not limited to "Form II" or "Form I" of nicotinic acid riboside triacetate (1-(2',3',5'-triacetyl-beta-D-ribofuranosyl)-nicotinic acid, "NAR triacetate", or NARTA), as disclosed in U.S. Pat. No. 11,214,589 and U.S. Pat. No. 10,689,411, respectively, and methods for their preparation.

[0052] Crystalline forms of useful molecules, also known as polymorphic crystal forms or "polymorphs", can have advantageous properties compared to the respective amorphous forms of such molecules. For example, crystalline forms are often easier to handle and process, for example, when preparing compositions containing the crystalline form. Crystalline forms typically have greater storage stability and are more amenable to purification. The use of crystalline forms of pharma- ceutical useful compounds can also improve the performance characteristics of pharmaceutical products containing the compound. Obtaining crystalline forms can also help formulation scientists to have a larger repertoire of materials available for formulation optimization, for example, by providing products with different properties, such as better processing or handling characteristics, improved dissolution profiles, or improved shelf life. Powder flow is important in formulation development for making tablets and capsules. Tabletting processes are based on powder volume and powder flow to maintain tablet weight uniformity. Therefore, designing the process and consistent control over powder flow properties is important to achieve optimal production. The development of a crystallization process that results in a form with novel physical and / or enhanced stability properties compared to inferior physical properties exhibited by other forms allows for formulation advances.

[0053] definition As used herein, the term "solvent" includes any of the following: water, water in which an ionic compound is dissolved, acetic acid, acetone, acetonitrile, benzene, 1-butanol, 2-butanol, t-butyl alcohol ("TBA", "t-BuOH"), 2-butanone, carbon tetrachloride, chlorobenzene, chloroform, cyclohexane, 1,2-dichloroethane ("DCE"), diethylene glycol, diethyl ether ("EtO"), diglyme (diethylene glycol dimethyl ether), 1,2-dimethoxyethane ("DME"), N,N-dimethylformamide ("DMF"), dimethylsulfoxide ("DMSO"), 1,4-dioxane, ethanol, ethyl acetate ("EtOAc"), ethylene glycol, glycerin, heptane, hexamethylphosphoramide ("HMPA") ... triamide ("HMPT"), hexane, methanol ("MeOH"), methyl t-butyl ether ("MTBE"), methylene chloride ("DCM", "CH2Cl2"), N-methyl-2-pyrrolidinone ("NMP"), nitromethane, pentane, petroleum ether, 1-propanol ("n-propanol", "n-PrOH"), 2-propanol ("isopropanol", "iPrOH"), pyridine, tetrahydrofuran ("THF"), toluene, triethylamine ("TEA", "Et3N"), o-xylene, m-xylene, and / or p-xylene. Solvent classes may include hydrocarbon, aromatic, aproptic, polar, alcohol, and mixtures thereof.

[0054] According to certain embodiments, the compounds or derivatives prepared according to embodiments of the methods of the present disclosure may include compounds or derivatives that are substantially free of solvents or other by-products, or that are typically free of certain solvents or by-products, or a salt, hydrate, solvate, or prodrug thereof, or a crystalline form thereof. In certain embodiments, "substantially free" means greater than about 80% by weight free of solvent or by-products, or greater than about 80% by weight free of a particular solvent or by-product, more preferably greater than about 90% by weight free of a solvent or by-product, or greater than about 90% by weight free of a particular solvent or by-product, even more preferably greater than about 95% by weight free of a solvent or by-product, or greater than about 95% by weight free of a particular solvent or by-product, even more preferably greater than 98% by weight free of a solvent or by-product, or greater than about 98% by weight free of a particular solvent or by-product, even more preferably greater than about 99% by weight free of a solvent or by-product, or greater than about 99% by weight free of a particular solvent or by-product, even more preferably greater than about 99.99% by weight free of a solvent or by-product, or greater than about 99.99% by weight free of a particular solvent or by-product, and most preferably quantitatively free of a solvent or by-product, or quantitatively free of a particular solvent or by-product.

[0055] For preparing pharmaceutical compositions from the crystal forms of nicotinamide riboside chloride or its hydrates, solvates, or prodrugs prepared according to the methods of the present disclosure, pharma- ceutically acceptable carriers can be solid or liquid.Solid form preparations include powders, tablets, pills, capsules, cachets, suppositories, and dispersible granules.Solid carriers can be one or more substances that can also function as diluents, flavoring agents, solubilizers, lubricants, suspending agents, binders, preservatives, tablet disintegrating agents, or encapsulating materials.

[0056] In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active component. In tablets, the active component is mixed with the carrier having the necessary binding capacity in suitable proportions and compacted to the desired shape and size.

[0057] The powders and tablets preferably contain from about 5 or 10 to about 70 percent of the active crystalline form of nicotinamide riboside (NR) or nicotinamide riboside triacetate (NRTA, VII) prepared according to the methods of the present disclosure, or a salt, hydrate, solvate, or prodrug thereof, such as the chloride salt (NRTA-Cl), or a mixture thereof. Suitable carriers include microcrystalline cellulose, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like, and other excipients may include magnesium stearate, stearic acid, talc, silicon dioxide, and the like. The dosage of the active form of nicotinamide riboside (NR) or nicotinamide riboside triacetate (NRTA, VII), or its salts, hydrates, solvates, or prodrugs, such as the chloride salt (NRTA-Cl), or mixtures thereof, may be, for example, about 10 mg to about 10,000 mg in the preparation. The term "preparation" is intended to include the formulation of the active compound with an encapsulating material as a carrier to provide a capsule in which the active ingredient, with or without a carrier, is surrounded by the carrier (and thus associated with the active ingredient). Tablets, powders, capsules, pills, sachets, and lozenges are included. Tablets, powders, capsules, pills, sachets, and lozenges can be used as solid forms suitable for oral administration.

[0058] Liquid preparations include solutions, suspensions, and emulsions, such as water or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as solutions in aqueous polyethylene glycol solutions. Thus, the crystal forms of nicotinamide riboside (NR) or nicotinamide riboside triacetate (NRTA, VII), or its salts, hydrates, solvates, or prodrugs, such as chloride salts (NRTA-Cl), or mixtures thereof, prepared according to the method of the present disclosure, can be formulated for parenteral administration (e.g., by injection, such as bolus injection or continuous infusion), and can be presented in unit doses, such as ampoules, pre-filled syringes, small volume injection solutions, or multi-dose containers with added preservatives. The compositions can take forms such as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulating agents, such as suspending agents, stabilizing agents, and / or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid from solution for reconstitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use, or by lyophilization.

[0059] The powders and tablets preferably contain about 1 to about 99.99 percent of the active crystalline form of nicotinamide riboside (NR, I) or nicotinamide riboside triacetate (NRTA, VII), or a salt, hydrate, solvate, or prodrug thereof, prepared according to the methods of the present disclosure. Suitable carriers include microcrystalline cellulose, sugar, lactose, pectin, dextrin, starch, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, low melting wax, cocoa butter, and the like, and other excipients may include magnesium stearate, stearic acid, talc, silicon dioxide, and the like. The dosage of the active form of nicotinamide riboside (NR, I) or nicotinamide riboside triacetate (NRTA, VII) may be, for example, about 10 mg to about 10,000 mg in the preparation. The term "preparation" is intended to include the formulation of the active compound with an encapsulating material as a carrier to provide a capsule in which the active ingredient, with or without a carrier, is surrounded by (and therefore associated with) the carrier. Tablets, powders, capsules, pills, sachets, and lozenges are included. Tablets, powders, capsules, pills, sachets, and lozenges can be used as solid forms suitable for oral administration. Liquid preparations include solutions, suspensions, and emulsions, for example, water or water-propylene glycol solutions. For example, parenteral injection liquid preparations can be formulated as a solution in aqueous polyethylene glycol solution. Thus, the crystalline forms of nicotinamide riboside (NR, I) or nicotinamide riboside triacetate (NRTA, VII), or salts, hydrates, solvates, or prodrugs thereof prepared according to the methods of the present disclosure, can be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion) and can be presented in unit doses, e.g., in ampoules, prefilled syringes, small volume injection solutions, or in multi-dose containers with added preservatives. The compositions can take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and can contain formulating agents, e.g., suspending, stabilizing, and / or dispersing agents.Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of sterile solid from a solution for reconstitution with a suitable vehicle, such as sterile pyrogen-free water, before use, or by lyophilization. Methods of administration may be via inhalation and topical routes. Aqueous solutions suitable for oral use can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavorings, stabilizers, and thickeners, if desired. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active ingredient in water with a viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0060] Compositions suitable for topical administration in the mouth include lozenges comprising the active ingredient in a flavored base, usually sucrose and gum arabic or gum tragacanth; pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin or sucrose and gum arabic; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

[0061] The solution or suspension is applied directly to the nasal cavity by conventional means, for example, by dropper, pipette or spray. The composition can be provided in single-dose or multi-dose form. In compositions intended for administration to the respiratory tract, including intranasal compositions, the compound or derivative usually has a small particle size, for example, on the order of 5 microns or less. Such a particle size can be obtained by means known in the art, for example, by micronization.

[0062] The pharmaceutical product is preferably in unit dosage form. In such form, the preparation is subdivided into unit doses containing appropriate amounts of active ingredients. The unit dosage form can be a packaged preparation, the package containing discrete amounts of the preparation, for example, tablets, capsules, and powders packaged in vials or ampoules. The unit dosage form can also be a capsule, tablet, cachet, or lozenge itself, or the appropriate number of any of these in packaged form.

[0063] Tablets, capsules, and lozenges for oral administration, and liquids for oral use are preferred compositions. Solutions or suspensions for application to the nasal cavity or to the respiratory tract are preferred compositions. Transdermal patches for topical administration to the epidermis are preferred compositions.

[0064] Aqueous solutions suitable for oral use can be prepared by dissolving the active ingredient in water and adding suitable colorants, flavors, stabilizers, and thickening agents, if desired. Aqueous suspensions suitable for oral use can be prepared by dispersing the finely divided active ingredient in water with a viscous material, such as natural or synthetic gums, resins, methylcellulose, sodium carboxymethylcellulose, or other well-known suspending agents.

[0065] The crystalline forms of beta nicotinamide riboside triacetate prepared by the methods of the present disclosure may take the form of a salt. The term "salt" includes addition salts of the free acid or free base of the crystalline forms of beta nicotinamide riboside triacetate prepared by the methods of the present disclosure. The term "pharmaceutically acceptable salt" refers to salts that have a toxicity profile within the range that makes them useful in pharmaceutical applications.

[0066] Further details regarding techniques for formulation can be found in the latest edition of Remington's Pharmaceutical Sciences (Mack Publishing Co., Easton, PA).

[0067] Additionally, embodiments of the present methods of treating and / or preventing signs, diseases, disorders, or conditions in a mammalian subject that are associated with or have an etiology involving vitamin B3 deficiency and / or that would benefit from increased mitochondrial activity address limitations of existing technologies for treating or preventing signs, diseases, disorders, or conditions that are associated with or have an etiology involving vitamin B3 deficiency and / or that would benefit from increased mitochondrial activity.

[0068] In certain embodiments, the present invention provides a method for treating and / or preventing signs, diseases, disorders, or symptoms associated with or associated with a pathology involving vitamin B3 deficiency.Exemplary signs, diseases, disorders, or symptoms associated with or associated with a pathology involving vitamin B3 deficiency that can be treated and / or prevented according to the described method include indigestion, fatigue, sore mouth, vomiting, poor circulation, burning mouth, red, swollen tongue, and depression.Severe vitamin B3 deficiency can cause premature aging symptoms characterized by a condition known as pellagra, cracked, scaly skin, dementia, and diarrhea.Other conditions characterized by premature or accelerated aging include Cockayne syndrome, Neil-Dingwall syndrome, progeria, etc.

[0069] In certain embodiments, the present invention provides a method for treating and / or preventing symptoms, diseases, disorders, or conditions that would benefit from increased mitochondrial activity.Increasing mitochondrial activity refers to increasing mitochondrial activity by increasing the number of mitochondria while simultaneously maintaining the overall number of mitochondria (e.g., mitochondrial mass), increasing mitochondrial activity (e.g., by stimulating mitochondrial biogenesis), or combinations thereof.In certain embodiments, symptoms, diseases, disorders, or conditions that would benefit from increased mitochondrial activity include symptoms, diseases, disorders, or conditions associated with mitochondrial dysfunction.

[0070] In certain embodiments, the method of treating and / or preventing the symptoms, diseases, disorders or conditions that would benefit from increased mitochondrial activity may include identifying subjects suffering from mitochondrial dysfunction. Methods of diagnosing mitochondrial dysfunction, which may involve molecular genetic analysis, pathological analysis, and / or biochemical analysis, are summarized in Bruce H. Cohen & Deborah R. Gold, Mitochondrial cytopathy in adults: what we know so far, 68 CLEVELAND CLINIC J. MED. 625 (2001). One method of diagnosing mitochondrial dysfunction is the Thor-Byrneier scale (see, for example, Cohen & Gold 2001; S. Collins et al., Respiratory Chain Encephalomyopathies: A Diagnostic Classification, 36 EUROPEAN NEUROLOGY 260 (1996)).

[0071] Mitochondria are important for the survival and proper function of almost all types of eukaryotic cells. Mitochondria in virtually any cell type may have congenital or acquired defects that affect their function. Thus, the clinically significant signs and symptoms of mitochondrial defects that affect respiratory chain function are heterogeneous and variable, depending on the distribution of defective mitochondria among cells and the severity of the defect, as well as on the physiological demands of the affected cells. Non-dividing tissues with high energy requirements, such as nervous tissue, skeletal muscle, and cardiac muscle, are particularly susceptible to mitochondrial respiratory chain dysfunction, but any organ system may be affected.

[0072] Signs, diseases, disorders, and conditions associated with mitochondrial dysfunction include those in which defects in mitochondrial respiratory chain activity contribute to the development of the pathophysiology of such signs, diseases, disorders, or conditions in a mammal, including 1) congenital genetic defects in the activity of one or more components of the mitochondrial respiratory chain, such defects being caused by a) oxidative impairment during aging; b) elevated intracellular calcium; c) exposure of affected cells to nitric oxide; d) hypoxia or ischemia; e) microtubule-associated defects in axonal transport of mitochondria; or f) expression of mitochondrial uncoupling proteins.

[0073] Indications, diseases, disorders, or conditions that would benefit from increased mitochondrial activity generally include, for example, diseases in which free radical-mediated oxidative damage causes tissue degeneration, diseases in which cells inappropriately undergo apoptosis, and diseases in which cells fail to undergo apoptosis. Exemplary indications, diseases, disorders, or conditions that would benefit from increased mitochondrial activity include, for example, AD (Alzheimer's disease), ADPD (Alzheimer's disease and Parkinson's disease), AMDF (ataxia, myoclonus, and hearing loss), autoimmune diseases, lupus, erythematosus, SLE (systemic lupus erythematosus), cataracts, cancer, CIPO (chronic intestinal pseudo-obstruction with myopathy and ophthalmoplegia), congenital muscular dystrophies, CPEO (chronic progressive external ophthalmoplegia), DEAF (maternal malformation, glaucoma, and chronic myocardial infarction), and glaucoma. Hereditary deafness or aminoglycoside-induced deafness), DEMCHO (dementia and chorea), diabetes mellitus (type I or type II), DID-MOAD (diabetes insipidus, diabetes mellitus, optic atrophy, deafness), DMDF (diabetes mellitus and deafness), ataxia, exercise intolerance, ESOC (epilepsy, stroke, optic atrophy, and cognitive decline), FBSN (familial bilateral striatal necrosis), FICP (fatal infantile cardiomyopathy plus, MELAS-associated cardiomyopathy), GER (gastroesophageal reflux disease), HD (Huntington's chorea) ), KSS (Kerns-Sayre syndrome), "late-onset" myopathy, LDYT (Leber's hereditary optic neuropathy and dystonia), Leigh's syndrome, LHON (Leber's hereditary optic neuropathy), LIMM (fatal infantile mitochondrial myopathy), MDM (muscle disease and diabetes mellitus), MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes), MEPR (myoclonic epilepsy and psychomotor regression), MERME (MERRF / MELAS overlap disorder), MERRF (myoclonus epilepsy and psychomotor regression), clonus epilepsy and ragged red fibers), MHCM (maternally inherited hypertrophic cardiomyopathy), MICM (maternally inherited cardiomyopathy), MILS (maternally inherited Leigh syndrome), mitochondrial encephalocardiopathy, mitochondrial encephalomyopathy, MM (mitochondrial myopathy), MMC (maternal myopathy and cardiomyopathy), MNGIE (myopathy and external ophthalmoplegia, neuropathy, gastrointestinal, encephalopathy), multisystem mitochondrial disorder (myopathy, encephalopathy, blindness, hearing loss, peripheral neuropathy), NARP (neurogenic weakness, ataxia, and retinitis pigmentosa;Alternative phenotypes at this locus include Leigh's disease, PD (Parkinson's disease), Pearson syndrome, PEM (progressive encephalopathy), PEO (progressive external ophthalmoplegia), PME (progressive myoclonic epilepsy), PMPS (Pearson medullary-pancreatic syndrome), psoriasis, RTT (Rett syndrome), schizophrenia, SIDS (sudden infant death syndrome), SNHL (sensory neurological hearing loss), various familial presentations (clinical manifestations range from spastic paraparesis to multisystem progressive disorder, from fatal cardiomyopathy to truncal ataxia, dysarthria, severe hearing loss, mental regression, ptosis, ophthalmoplegia, distal cycloplegia, and diabetes mellitus), or Wolfram syndrome;

[0074] Other indications, diseases, disorders, and conditions that would benefit from increased mitochondrial activity include, for example, Friedreich's ataxia and other ataxias, amyotrophic lateral sclerosis (ALS) and other motor neuron diseases, macular degeneration, epilepsy, Alpers syndrome, multiple mitochondrial DNA deletion syndrome, MtDNA depletion syndrome, complex I deficiency, complex II (SDH) deficiency, complex III deficiency, cytochrome c oxidase (COX, complex IV) deficiency, complex V deficiency, adenine nucleotide translocator (ANT) deficiency, pyruvate dehydrogenase (PDH) deficiency, ethylmalonic aciduria with lactic acidemia, refractory glaucoma with weakness during infection, and chronic kidney disease. epilepsy, Asperger's syndrome with weakness during infection, autism with weakness during infection, attention deficit hyperactivity disorder (ADHD), cerebral palsy with weakness during infection, dyslexia with weakness during infection, materially inherited thrombocytopenia and leukemia syndrome, MARIAHS syndrome (mitochondrial ataxia, recurrent infections, aphasia, hypouricemia / hypomyelination, seizures, and dicarboxylic aciduria), ND6 ataxia, cyclic vomiting syndrome with weakness during infection, 3-hydroxyisobutyric aciduria with lactic acidemia, diabetes mellitus with lactic acidemia, uridine responsive neurological syndrome (URNS), dilated cardiomyopathy, splenic lymphoma, or renal tubular acidosis / diabetes / ataxia syndrome.

[0075] In other embodiments, the present invention provides methods of treating a mammal (e.g., a human) suffering from mitochondrial disorders resulting from, but not limited to, post-traumatic head injury and cerebral edema, stroke (methods of the invention useful for treating or preventing reperfusion injury), Lewy body dementia, hepatorenal syndrome, acute liver failure, NASH (non-alcoholic steatohepatitis), anti-metastasis / prodifferentiation therapy of cancer, idiopathic congestive heart failure, atrial fibrillation (non-valvular), Wolff-Parkinson-White syndrome, idiopathic heart block, prevention of reperfusion injury in acute myocardial infarction, familial migraine, irritable bowel syndrome, secondary prevention of non-Q wave myocardial infarction, premenstrual syndrome, prevention of renal failure in hepatorenal syndrome, antiphospholipid syndrome, eclampsia / preeclampsia, oopause infertility, ischemic heart disease / angina, and Shy-Drager and unclassifiable dysautonomia syndromes.

[0076] In yet another embodiment, a method for treating mitochondrial disorders with pharmacological drug-related side effects is provided. Types of pharmacological agents associated with mitochondrial disorders include reverse transcriptase inhibitors, protease inhibitors, DHOD inhibitors, etc. Examples of reverse transcriptase inhibitors include, for example, azidothymidine (AZT), stavudine (D4T), zalcitabine (ddC), didanosine (DDI), fluoroiodoarauracil (FIAU), lamivudine (3TC), abacavir, etc. Examples of protease inhibitors include, for example, ritonavir, indinavir, saquinavir, nelfinavir, etc. Examples of dihydroorotate dehydrogenase (DHOD) inhibitors include, for example, leflunomide, brequinar, etc.

[0077] Reverse transcriptase inhibitors not only inhibit reverse transcriptase, but also polymerase gamma, which is required for mitochondrial function. Thus, inhibition of polymerase gamma activity (e.g., by reverse transcriptase inhibitors) causes mitochondrial dysfunction and / or reduced mitochondrial mass, which manifests itself in patients as hyperlactatemia. Conditions of this type may benefit from increased mitochondrial number and / or improved mitochondrial function.

[0078] Common symptoms of mitochondrial diseases include cardiomyopathy, muscle weakness and atrophy, developmental delays (involving motor, language, cognitive, or executive functions), ataxia, epilepsy, renal tubular acidosis, peripheral neuropathy, optic neuropathy, autonomic nervous system neuropathy, neurogenic bowel dysfunction, sensorineural hearing loss, neurogenic bladder dysfunction, dilated cardiomyopathy, migraine headaches, liver failure, lactic acidemia, and diabetes mellitus.

[0079] EMBODIMENTS OF THE PRESENT DISCLOSURE The cited invention provides a scalable crystallization process for producing crystalline beta-nicotinamide riboside triacetate chloride. The improved process features described herein produce crystalline material with low residual solvent content, large crystal particle size, narrow particle size distribution, and high yield suitable for use as a commercial dietary supplement.

[0080] The novel components of the present invention include: improved crystal size and particle size distribution relative to other crystallization processes; low residual solvent content relative to crystalline material produced via other crystallization processes; and improved yield relative to other crystallization processes. The process described herein above achieves the preparation of said crystalline beta nicotinamide riboside triacetate chloride.

[0081] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride may include the steps disclosed in U.S. Pat. No. 9,975,915, which is incorporated herein by reference in its entirety.

[0082] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride includes: a) adding an amount of crude nicotinamide riboside triacetate chloride to a vessel at a first temperature, optionally wherein the amount of crude nicotinamide riboside triacetate chloride is between about 65 g and about 80 g and the first temperature is between about 18° C. and about 23° C.; b) adding an amount of water and an amount of ethanol to form a reaction mixture, where in some cases the amount of water is about 75 g to about 90 g and the amount of ethanol is about 38 g to about 50 g; c) stirring and heating the reaction mixture to a second temperature, optionally wherein the second temperature is from about 48° C. to about 52° C.; d) cooling the vessel once at the second temperature to a third temperature, optionally wherein the third temperature is from about 28° C. to about 32° C.; e) slowly metering an amount of ethanol into the container at a first rate, optionally wherein the amount of ethanol is between about 850 g and about 950 g and the first rate is between about 8 mL / min and about 12 mL / min; f) maintaining the container at a third temperature for a first period of time, optionally the first period of time being from about 45 minutes to about 75 minutes; g) cooling the container to a fourth temperature, optionally the fourth temperature being from about −8° C. to about −12° C., and holding the fourth temperature for a second period of time, optionally the second period of time being from about 8 hours to about 16 hours; h) observing crystal formation at or before the fifth temperature, optionally wherein the fifth temperature is from about -5°C to about -9°C; i) obtaining nicotinamide riboside triacetate chloride as a white, crystalline powder; may include.

[0083] The crystalline forms of nicotinamide riboside triacetate chloride of the present disclosure can be isolated from the reaction mixture and purified by standard techniques, such as filtration, liquid-liquid extraction, solid phase extraction, distillation, recrystallization, or chromatography, including flash column chromatography, preparative TLC, HPTLC, HPLC, or rp-HPLC. A preferred method for the preparation of the crystalline forms of nicotinamide riboside triacetate chloride of the present disclosure includes crystallizing the compound or its salt, hydrate, solvate, or prodrug from a solvent, preferably to form a crystalline form of the compound or its derivative, or its salt, hydrate, solvate, or prodrug. After crystallization, the crystallization solvent is removed by a process other than evaporation, such as filtration or decantation, and the crystals are then preferably washed with a pure solvent (or a mixture of pure solvents). Preferred solvents for crystallization include water; alcohols, especially those containing up to four carbon atoms, such as methanol, ethanol, isopropanol, butan-1-ol, butan-2-ol, and 2-methyl-2-propanol; ethers, such as diethyl ether, diisopropyl ether, t-butyl methyl ether, 1,2-dimethoxyethane, tetrahydrofuran, and 1,4-dioxane; carboxylic acids, such as formic acid and acetic acid; hydrocarbon solvents, such as pentane, hexane, and toluene; and mixtures thereof, especially aqueous mixtures, such as aqueous methanol, ethanol, isopropanol, and acetone. Pure solvents, preferably at least analytical grade, more preferably pharmaceutical grade, are preferably used. In a preferred embodiment of the method of the present invention, the crystalline form is so isolated. As described above, the solvate of crystalline NRTA chloride may comprise one or more of the above solvents.

[0084] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride includes: a) adding an amount of crude nicotinamide riboside triacetate chloride to a vessel at a first temperature, optionally wherein the amount of crude nicotinamide riboside triacetate chloride is between about 65 g and about 80 g and the first temperature is between about 18° C. and about 23° C.; b) adding an amount of water and an amount of ethanol to form a reaction mixture, where in some cases the amount of water is about 75 g to about 90 g and the amount of ethanol is about 38 g to about 50 g; c) stirring and heating the reaction mixture to a second temperature, optionally wherein the second temperature is from about 28° C. to about 32° C.; d) slowly metering an amount of ethanol into the container at a first rate, optionally wherein the amount of ethanol is between about 850 g and about 950 g and the first rate is between about 8 mL / min and about 12 mL / min; e) cooling the vessel to a third temperature, optionally the third temperature being from about −8° C. to about −12° C., and holding the fourth temperature for a first period of time, optionally the first period of time being from about 8 hours to about 16 hours; f) observing crystal formation at or before the fourth temperature, optionally wherein the fourth temperature is from about -5°C to about -9°C; g) obtaining nicotinamide riboside triacetate chloride as a white, crystalline powder; may include.

[0085] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride includes: a) adding an amount of crude nicotinamide riboside triacetate chloride to a vessel at a first temperature, optionally wherein the amount of crude nicotinamide riboside triacetate chloride is from about 650 g to about 550 g and the first temperature is from about 18° C. to about 23° C.; b) adding an amount of water and an amount of ethanol to form a reaction mixture, where in some cases the amount of water is about 305 g to about 450 g and the amount of ethanol is about 150 g to about 250 g; c) stirring and heating the reaction mixture to a second temperature, optionally wherein the second temperature is from about 28° C. to about 32° C.; d) slowly metering an amount of ethanol into the container at a first rate, optionally wherein the amount of ethanol is between about 875 g and about 975 g and the first rate is between about 25 mL / min and about 35 mL / min; e) cooling the vessel to a third temperature, optionally wherein the third temperature is from about 8° C. to about 12° C.; f) seeding the crystals with an amount of nicotinamide riboside triacetate chloride, optionally wherein the amount of nicotinamide riboside triacetate chloride is from about 4 g to about 10 g; g) allowing the isotherm for a first period of time, optionally the first period of time being from about 30 minutes to about 90 minutes; h) slowly adding an amount of ethanol at a second rate, optionally wherein the amount of ethanol is between about 2100 g and about 2300 g and the second rate is between about 10 mL / min and about 20 mL / min; i) cooling the container to a fourth temperature in a second period of time, optionally wherein the fourth temperature is from about −5° C. to about 5° C. and the fourth temperature is from about 40 minutes to about 60 minutes; j) cooling the container to a fifth temperature in a third period of time, optionally wherein the fifth temperature is from about −15° C. to about 25° C. and the third period of time is from about 15 minutes to about 30 minutes; obtaining nicotinamide riboside triacetate chloride as a white, crystalline powder; may include.

[0086] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride includes: a) adding an amount of crude nicotinamide riboside triacetate chloride to a vessel at a first temperature, optionally wherein the amount of crude nicotinamide riboside triacetate chloride is between about 110 g and about 130 g and the first temperature is between about 18° C. and about 23° C.; b) adding an amount of water and an amount of ethanol to form a reaction mixture, where in some cases the amount of water is about 65 g to about 80 g and the amount of ethanol is about 38 g to about 58 g; c) stirring and heating the reaction mixture to a second temperature, optionally wherein the second temperature is from about 28° C. to about 32° C.; d) slowly metering an amount of ethanol into the container at a first rate, optionally wherein the amount of ethanol is between about 150 g and about 190 g and the first rate is between about 8 mL / min and about 12 mL / min; e) cooling the vessel to a third temperature, optionally wherein the third temperature is from about 8° C. to about 15° C.; f) seeding the crystals by adding an amount of nicotinamide riboside triacetate chloride, optionally wherein the amount of nicotinamide riboside triacetate chloride is about 1.0 g to 1.5 g; g) observing an isotherm for a first period of time, optionally the first period of time being from about 45 minutes to about 90 minutes; h) slowly adding an amount of ethanol at a second rate, optionally wherein the amount of ethanol is between about 400 g and about 475 g and the second rate is between about 2 mL / min and about 9 mL / min; g) cooling the container to a fourth temperature for a second period of time, optionally wherein the fourth temperature is from about −5° C. to about 5° C. and the second period of time is from about 225 minutes to about 275 minutes; i) cooling the container to a fifth temperature for a third period of time, optionally wherein the fifth temperature is from about −10° C. to about 0° C. and the third period of time is from about 40 minutes to about 60 minutes; j) cooling the container to a sixth temperature for a fourth period of time, optionally wherein the sixth temperature is from about −15° C. to about −5° C. and the fourth period of time is from about 10 minutes to about 30 minutes; k) cooling the vessel to a seventh temperature for a fifth period of time, optionally wherein the seventh temperature is from about −15° C. to about −5° C. and the fifth period of time is from about 8 hours to about 16 hours; l) obtaining nicotinamide riboside triacetate chloride as a white, crystalline powder; may include.

[0087] In one embodiment, a method for producing crystalline nicotinamide riboside triacetate chloride includes: a) adding an amount of crude nicotinamide riboside triacetate chloride to a vessel at a first temperature, optionally wherein the amount of crude nicotinamide riboside triacetate chloride is between about 110 g and about 130 g and the first temperature is between about 18° C. and about 23° C.; b) adding an amount of water and an amount of ethanol to form a reaction mixture, where in some cases the amount of water is about 65 g to about 80 g and the amount of ethanol is about 38 g to about 58 g; c) stirring and heating the reaction mixture to a second temperature, optionally wherein the second temperature is from about 28° C. to about 32° C.; d) slowly metering an amount of ethanol into the container at a first rate, optionally wherein the amount of ethanol is between about 150 g and about 190 g and the first rate is between about 8 mL / min and about 12 mL / min; e) cooling the vessel to a third temperature, optionally wherein the third temperature is from about 10° C. to about 20° C.; f) seeding the crystals by adding an amount of nicotinamide riboside triacetate chloride, optionally wherein the amount of nicotinamide riboside triacetate chloride is about 1.0 g to 1.5 g; g) observing an isotherm for a first period of time, optionally the first period of time being from about 45 minutes to about 90 minutes; h) slowly adding an amount of ethanol at a second rate, optionally wherein the amount of ethanol is between about 400 g and about 475 g and the second rate is between about 2 mL / min and about 9 mL / min; g) cooling the container to a fourth temperature for a second period of time, optionally wherein the fourth temperature is from about −5° C. to about 5° C. and the second period of time is from about 275 minutes to about 350 minutes; i) cooling the container to a fifth temperature for a third period of time, optionally wherein the fifth temperature is from about −10° C. to about 0° C. and the third period of time is from about 40 minutes to about 60 minutes; j) cooling the container to a sixth temperature for a fourth period of time, optionally wherein the sixth temperature is from about −15° C. to about −10° C. and the fourth period of time is from about 10 minutes to about 30 minutes; k) cooling the vessel to a seventh temperature for a fifth period of time, optionally wherein the seventh temperature is from about −15° C. to about −5° C. and the fifth period of time is from about 8 hours to about 16 hours; l) obtaining nicotinamide riboside triacetate chloride as a white, crystalline powder; may include.

[0088] In one embodiment, the crystalline nicotinamide riboside triacetate chloride may be characterized by a particle size distribution including 0.07% greater than 850 μm in size, 13.92% between 850 and 425 μm in size, 47.78% between 425 and 250 μm in size, 33.86% between 250 and 180 μm in size, 0.29% between 180 and 150 μm in size, 2.05% between 150 and 125 μm in size, 1.16% between 125 and 75 μm in size, and 0.55% between 75 and 0 μm in size.

[0089] In one embodiment, the crystalline nicotinamide riboside triacetate chloride may be characterized by a particle size distribution including 0.04% greater than 850 μm in size, 5.53% between 850 and 425 μm in size, 39.32% between 425 and 250 μm in size, 26.51% between 250 and 180 μm in size, 9.43% between 180 and 150 μm in size, 9.59% between 150 and 125 μm in size, 7.47% between 125 and 75 μm in size, and 1.61% between 75 and 0 μm in size.

[0090] In one embodiment, the crystalline nicotinamide riboside triacetate chloride may be characterized by a particle size distribution including 2.52% greater than 850 μm in size, 44.76% between 850 and 425 μm in size, 16.86% between 425 and 250 μm in size, 11.16% between 250 and 180 μm in size, 6.92% between 180 and 150 μm in size, 8.14% between 150 and 125 μm in size, 7.76% between 125 and 75 μm in size, and 1.69% between 75 and 0 μm in size.

[0091] In one embodiment, the crystalline nicotinamide riboside triacetate chloride may be characterized by a particle size distribution including 0.91% greater than 850 μm in size, 21.57% between 850 and 425 μm in size, 22.95% between 425 and 250 μm in size, 21.57% between 250 and 180 μm in size, 9.46% between 180 and 150 μm in size, 10.27% between 150 and 125 μm in size, 10.20% between 125 and 75 μm in size, and 2.86% between 75 and 0 μm in size.

[0092] In one embodiment, the crystalline nicotinamide riboside triacetate chloride may be characterized by a particle size distribution including 1.14% greater than 850 μm in size, 36.68% between 850 and 425 μm in size, 18.08% between 425 and 250 μm in size, 16.77% between 250 and 180 μm in size, 9.86% between 180 and 150 μm in size, 10.95% between 150 and 125 μm in size, 5.77% between 125 and 75 μm in size, and 0.17% between 75 and 0 μm in size.

[0093] The above-mentioned method can be further understood in conjunction with the following examples. In addition, the following non-limiting examples are provided to illustrate the present invention. The preparation procedures specifically described are applicable to other embodiments of the present invention. The preparation procedures described as general methods are stated as being typically effective to carry out the preparations shown. However, those skilled in the art will recognize that it may be necessary to vary the procedure for any given embodiment of the present invention, for example, to vary the order or steps and / or reagents used. The product may be purified by conventional techniques, but this will vary according to, for example, the physical properties of the crystalline form prepared according to the method of the present invention. EXAMPLES

[0094] Working Example The following non-limiting examples are provided to illustrate the present invention, however, one of ordinary skill in the art will recognize that it may be necessary to vary the procedures for any given embodiment of the invention, such as varying the order or steps of the methods.

[0095] Example 1: NRTA-Cl synthesis Preparation of Nicotinamide Riboside Triacetate Chloride Nicotinamide riboside triacetate chloride (NRTA-Cl) can be prepared as disclosed in U.S. Patent No. 9,975,915 and U.S. Patent No. 10,689,411, which are incorporated herein by reference in their entireties. In an alternative preparation, a 5 L jacketed reactor was charged with 1034 g (3.25 mol) of β-D-ribofuranose 1,2,3,5-tetraacetate and 1392 g of CH3CN. The mixture was stirred at 20°C until dissolved. After dissolution, the reactor was cooled to -10°C. At this point, 13 g (0.16 mol, 0.05 equiv.) of acetyl chloride was charged. The reactor was further cooled to -15°C. At this point, 146 g (4.06 mol, 1.25 eq) of anhydrous hydrogen chloride gas was sparged into the reaction mixture at 1.5 g / min while maintaining an internal temperature below -8°C. After charging, the reaction was allowed to sit at -15°C overnight. After the -15°C isotherm, 555 g (4.55 mol, 1.40 eq) of nicotinamide was charged into the reactor along with 757 g of CH3CN. The mixture was stirred at -15°C for 2 hours and then warmed to 20°C and held overnight. After the 20°C isotherm, the reaction was cooled to -5°C. At this point, 602.39 g (3.25 mol, 1.00 eq) of tributylamine was slowly charged into the reactor. The reactor was warmed to 23°C and stirred for 3 hours. The resulting crude material, nicotinamide-D-ribofuranose triacetate chloride, was washed with 2500 mL CH3CN and dried under vacuum at 40° C. 694 g of a light white, crystalline powder was obtained (51% yield).

[0096] Purity as determined by HPLC: 100.70% nicotinamide riboside triacetate chloride, 0.513% nicotinamide.

[0097] Residual solvents by GC-MS: Ethanol at undetectable ppm, Acetonitrile at 3392.553 ppm.

[0098] Example 2: NRTA-Cl Crystallization, Self-Seeding Preparation of Nicotinamide Riboside Triacetate Chloride Crystals Crude nicotinamide riboside triacetate chloride product (about 78 g), produced similarly to Example 1, was added to a 1 L jacketed reactor set at about 20° C. To the crude product was added about 83 g of water and about 42 g of ethanol. The resulting mixture was stirred and heated to about 50° C. to aid in dissolution. Once at about 50° C., the reactor was cooled to about 30° C. At this point, about 907 g of additional ethanol was slowly metered into the reactor at about 10 mL / min. After the ethanol addition, the reactor was held at about 30° C. for 1 hour. The reactor was then cooled to about −10° C. and held overnight, or for about 8 to about 12 hours. In this method, crystal formation was initially observed at about −7° C. About 54.5 g of dry white, crystalline powder was obtained (about 70% mass recovery, about 76% yield (adjusted for dry content and starting material purity)).

[0099] The crystalline nicotinamide riboside triacetate chloride contained approximately 99.6% purity as determined by HPLC. The nicotinamide riboside triacetate chloride contained approximately 0.222% nicotinamide.

[0100] Nicotinamide riboside triacetate chloride contained residual solvents as determined by GC-MS: approximately 3392.35 ppm ethanol, less than approximately 10 ppm undetectable acetonitrile.

[0101] Nicotinamide riboside triacetate chloride contained the following particle sizes as determined by sieve analysis: >850μm-0.07%, about 850-425μm-13.92%, about 425-250μm-47.78%, about 250-180μm-33.86%, about 180-150μm-0.29%, about 150-125μm-2.05%, about 125-75μm-1.16%, about 75-0μm-0.55%. Bulk density was measured using USA Method 786, Stage 6 Harmonization, Official August 1, 2015.

[0102] Example 3: NRTA-Cl Crystallization, Self-Seeding Preparation of Nicotinamide Riboside Triacetate Chloride Crystals Crude nicotinamide riboside triacetate chloride product (about 78 g), produced similarly to Example 1, was added to a 1 L jacketed reactor set at about 20° C. To the crude product, about 83 g of water and about 42 g of ethanol were added. The resulting mixture was stirred and heated to about 30° C. to aid in dissolution. Once in solution, about 907 g of additional ethanol was slowly metered into the reactor at about 10 mL / min. The reactor was then cooled to about −10° C. and held overnight or for about 8 to about 12 hours. In this method, crystal formation was first observed at about −7° C. About 61 g of white, crystalline powder dry matter (about 78% mass recovery, about 85% yield (adjusted for dry content and starting material purity)) was obtained.

[0103] The crystalline nicotinamide riboside triacetate chloride contained a purity of about 99.3% as determined by HPLC. The nicotinamide riboside triacetate chloride contained a BRL of <0.17% nicotinamide.

[0104] Nicotinamide riboside triacetate chloride contained residual solvents as determined by GC-MS: approximately 3726.08 ppm ethanol, no detectable acetonitrile.

[0105] Nicotinamide riboside triacetate chloride contained the following particle sizes as determined by sieve analysis: greater than 850 μm-0.04%, about 850-425 μm-5.53%, about 425-250 μm-39.32%, about 250-180 μm-26.51%, about 180-150 μm-9.43%, about 150-125 μm-9.59%, about 125-75 μm-7.47%, and about 75-0 μm-1.61%.

[0106] Example 4: NRTA-Cl crystallization, seeding Preparation of Nicotinamide Riboside Triacetate Chloride Crystals Crude nicotinamide riboside triacetate chloride product (about 577 g) produced similar to Example 1 was added to a 5 L jacketed reactor set at about 20° C. To the crude product was added about 373 g water and about 191 g ethanol. The resulting mixture was stirred and heated to about 30° C. to aid in dissolution. Once in solution, about 925 g additional ethanol was slowly metered into the reactor at about 30 mL / min. The reactor was then cooled to about 10° C. Once at about 10° C., about 6 g of nicotinamide riboside triacetate chloride seed crystals produced similar to Example 3 were added into the reactor and an isotherm was allowed to run for about 1 hour. Crystal formation was observed immediately after seed addition. After about a 1 hour isotherm, an additional about 2,229 g of ethanol was slowly metered into the reactor at about 15 mL / min. At the start of the second addition of ethanol, the reactor was first cooled to about 0° C. over about 200 minutes, then to about −5° C. over about 50 minutes, and finally to about −10° C. over about 20 minutes. The reactor was then cooled to about −10° C. and held overnight or for about 8 to about 12 hours. The seed material was removed to give about 469 g of dry matter of a white, crystalline powder (about 81% mass recovery, about 83% yield (adjusted for starting material dry content and purity)).

[0107] The crystalline nicotinamide riboside triacetate chloride contained approximately 100.7% purity as determined by HPLC. The nicotinamide riboside triacetate chloride contained approximately 0.513% nicotinamide.

[0108] Nicotinamide riboside triacetate chloride contained residual solvents as determined by GC-MS: approximately 538.799 ppm ethanol, no detectable acetonitrile.

[0109] Nicotinamide riboside triacetate chloride contained the following particle size distribution as determined by sieve analysis: greater than 850 μm-2.52%, about 850-425 μm-44.76%, about 425-250 μm-16.86%, about 250-180 μm-11.16%, about 180-150 μm-6.92%, about 150-125 μm-8.14%, about 125-75 μm-7.76%, and about 75-0 μm-1.69%.

[0110] Example 5: NRTA-Cl crystallization, seeding Preparation of Nicotinamide Riboside Triacetate Chloride Crystals Crude nicotinamide riboside triacetate chloride product (about 121 g) produced similar to Example 1 was added to a 1 L jacketed reactor set at about 20° C. To the crude product was added about 72 g water and about 48 g ethanol. The resulting mixture was stirred and heated to about 30° C. to aid in dissolution. Once in solution, about 168 g additional ethanol was slowly metered into the reactor at about 10 mL / min. The reactor was then cooled to about 12.5° C. Once at about 12.5° C., about 1.2 g of nicotinamide riboside triacetate chloride seed crystals produced similar to Example 3 were added into the reactor and an isotherm was allowed to run for about 1 hour. Crystal formation was observed immediately after seed addition. After about a 1 hour isotherm, an additional about 431 g of ethanol was slowly metered into the reactor at about 5 mL / min. At the start of the second addition of ethanol, the reactor was first cooled to about 0° C. over about 250 minutes, then to about −5° C. over about 50 minutes, and finally to about −10° C. over about 20 minutes. The reactor was then cooled to about −10° C. and held overnight or for about 8 to about 12 hours. The seed material was removed to give about 104.5 g of dry white, crystalline powder (about 86% mass recovery, about 89% yield (adjusted for starting material dry content and purity)).

[0111] The crystalline nicotinamide riboside triacetate chloride contained approximately 99.8% purity as determined by HPLC. The nicotinamide riboside triacetate chloride contained approximately 0.348% nicotinamide.

[0112] Nicotinamide riboside triacetate chloride contained residual solvents as determined by GC-MS: approximately 294.615 ppm ethanol, no detectable acetonitrile.

[0113] Nicotinamide riboside triacetate chloride contained the following particle sizes as determined by sieve analysis: greater than 850 μm-0.91%, about 850-425 μm-21.57%, about 425-250 μm-22.95%, about 250-180 μm-21.57%, about 180-150 μm-9.46%, about 150-125 μm-10.27%, about 125-75 μm-10.20%, and about 75-0 μm-2.86%.

[0114] Example 6: NRTA-Cl Crystallization, Seeding Preparation of Nicotinamide Riboside Triacetate Chloride Crystals Crude nicotinamide riboside triacetate chloride product (about 121 g) produced similar to Example 1 was added to a 1 L jacketed reactor set at about 20° C. To the crude product was added about 72 g water and about 48 g ethanol. The resulting mixture was stirred and heated to about 30° C. to aid in dissolution. Once in solution, about 168 g additional ethanol was slowly metered into the reactor at about 10 mL / min. The reactor was then cooled to about 15° C. Once at about 15° C., about 1.2 g of nicotinamide riboside triacetate chloride seed crystals produced similar to Example 3 were added into the reactor and an isotherm was allowed to run for about 1 hour. Crystal formation was observed immediately after seed addition. After the 1 hour isotherm, an additional about 431 g of ethanol was slowly metered into the reactor at about 5 mL / min. At the start of the second addition of ethanol, the reactor was first cooled to about 0° C. over about 300 minutes, then to about −5° C. over about 50 minutes, and finally to about −10° C. over about 20 minutes. The reactor was then cooled to about −10° C. and held overnight or for about 8 to about 12 hours. The seed material was removed to give about 103.9 g of dry matter of a white, crystalline powder (about 86% mass recovery, about 88% yield (adjusted for starting material dry content and purity)).

[0115] The crystalline nicotinamide riboside triacetate chloride contained approximately 99.2% purity as determined by HPLC. The nicotinamide riboside triacetate chloride contained approximately 0.351% nicotinamide.

[0116] Nicotinamide riboside triacetate chloride contained residual solvents as determined by GC-MS: approximately 339.802 ppm ethanol, no detectable acetonitrile.

[0117] Nicotinamide riboside triacetate chloride contained the following particle sizes as determined by sieve analysis: greater than 850 μm-1.14%, about 850-425 μm-36.68%, about 425-250 μm-18.08%, about 250-180 μm-16.77%, about 180-150 μm-9.86%, about 150-125 μm-10.95%, about 125-75 μm-5.77%, and about 75-0 μm-0.17%.

[0118] The use of the terms "a", "an", "the" and similar referents in the context of describing the claimed invention (especially in the context of the claims) should be construed to cover both the singular and the plural, unless otherwise specified herein or clearly contradicted by context. The description of ranges of values ​​herein is merely intended to serve as a shorthand for individually referring to each separate value falling within the range, unless otherwise specified herein, and each separate value is incorporated herein as if individually recited herein. The use of the term "about" is intended to describe values ​​that are greater than or less than the stated value within approximately ±10%; in other embodiments, values ​​may range to values ​​that are greater than or less than the stated value within approximately ±5%; in other embodiments, values ​​may range to values ​​that are greater than or less than the stated value within approximately ±2%; in other embodiments, values ​​may range to values ​​that are greater than or less than the stated value within approximately ±1%. The preceding ranges are intended to be made clear by the context, and no further limitations are included. All methods described herein may be performed in any suitable order unless otherwise specified herein or clearly contradicted by the context. The use of any and all examples or exemplary language provided herein (e.g., "such as") is intended merely to better clarify the invention, and does not pose a limitation on the scope of the invention unless specifically claimed. Language in the present specification should not be construed as indicating any non-claimed element essential to the practice of the invention.

[0119] In the foregoing specification, the invention has been described with respect to specific embodiments thereof, and numerous details have been set forth for purposes of illustration, but it will be apparent to those skilled in the art that the invention is susceptible to additional embodiments, and that the details described herein may be varied considerably without departing from the basic principles of the invention.

[0120] All references cited herein are incorporated by reference in their entirety. Since the invention may be embodied in other specific forms without departing from its spirit or essential attributes, reference should be made to the appended claims, rather than the foregoing specification, as indicating the scope of the invention.

Claims

1. 1. A substantially crystalline compound having the formula (VII), or a salt or solvate thereof: 【Chemistry 1】 In the formula, X － is a counter ion; A compound having a chemical purity of greater than about 90% (w / w) and containing less than about 5000 ppm ethanol.

2. 2. The compound of claim 1, substantially in the beta anomeric form.

3. X － are fluoride ion, chloride ion, bromide ion, iodide ion, formate ion, acetate ion, propionate ion, butyrate ion, glutamate ion, aspartate ion, ascorbate ion, benzoate ion, carbonate ion, citrate ion, carbamate ion, gluconate ion, lactate ion, nitrate ion, phosphate ion, diphosphate ion, sulfate ion, succinate ion, sulfonate ion, trifluoromethanesulfonate ion, trichloromethanesulfonate ion, tribromomethanesulfonate ion, trichloroacetate ion, tribromoacetate ion, trifluoroacetate ion, malate ion, hydrogen malate ion, tartrate ion, hydrogen tartrate ion, glycolate ion, glucuronate ion, maleate ion, fumarate ion, 2. The compound of claim 1, wherein the anion is selected from the group consisting of carboxylate, pyruvate, anthranilate, 4-hydroxybenzoate, phenylacetate, mandelate, pamoate, methanesulfonate, ethanesulfonate, benzenesulfonate, pantothenate, 2-hydroxyethanesulfonate, p-toluenesulfonate, sulfanilate, cyclohexylaminosulfonate, stearate, palmitate, myristate, laurate, caprate, caprylate, caproate, oleate, linoleate, alginate, beta-hydroxybutyrate, salicylate, galactarate, and galacturonate.

4. X － The compound of claim 2, wherein is a chloride salt having Form I.

5. 5. The compound of claim 4 containing less than about 1000 ppm ethanol.

6. A nutritional supplement comprising any one of claims 1 to 5, which comprises an excipient or carrier.

7. A pharmaceutical composition comprising any one of claims 1 to 5 and a pharma- ceutically acceptable carrier.

8. 10. A method for producing the substantially crystalline compound nicotinamide riboside triacetate or a salt or solvate thereof of claim 1, comprising: (a) adding an amount of crude nicotinamide riboside triacetate to a volume of a first solvent to form a reaction mixture; (b) heating the reaction mixture to a temperature of about 20° C. to about 60° C.; (c) cooling the reaction mixture; (d) adding a second solvent; (e) isolating the substantially crystalline compound nicotinamide riboside triacetate or a salt or solvate thereof as a crystalline powder. The method includes:

9. (b1) adding a third solvent immediately after step (b); The method of claim 8 , further comprising:

10. (c1) after step (c), seeding the reaction mixture with the crystalline compound nicotinamide riboside triacetate or a salt or solvate thereof; The method of claim 8 , further comprising:

11. 11. The method of claim 10, wherein the crystalline compound nicotinamide riboside triacetate is a chloride salt having Form I.

12. 9. The method of claim 8, wherein the solvate of nicotinamide riboside comprises a solvent selected from the group consisting of water, acetic acid, acetone, acetonitrile, 1-butanol, 2-butanol, t-butyl alcohol, cyclohexane, 1,2-dichloroethane, diethylene glycol, diethyl ether, diglyme (diethylene glycol dimethyl ether), 1,2-dimethoxyethane, N,N-dimethylformamide, dimethylsulfoxide, 1,4-dioxane, ethanol, ethyl acetate, ethylene glycol, methanol ("MeOH"), methyl t-butyl ether, N-methyl-2-pyrrolidinone, 1-propanol, 2-propanol, pyridine, and tetrahydrofuran.

13. The method of claim 8 , wherein the first solvent is selected from water, ethanol, or a mixture thereof.

14. 9. The method of claim 8, wherein the second solvent is selected from one or more solvents selected from the group consisting of methyl t-butyl ether, acetone, methanol, ethanol, acetonitrile, and water.

15. 12. The method of claim 11, wherein the crystalline nicotinamide riboside triacetate chloride Form I has a chemical purity of at least 99% as determined by HPLC.

16. A method for treating and / or preventing a condition associated with or having an etiology involving vitamin B3 deficiency in a subject mammal, comprising administering to said subject a therapeutically effective amount of intracellular NADPH, selected from the group consisting of indigestion, fatigue, sores in the mouth, vomiting, poor circulation, burning mouth, swollen red tongue, depression, pellagra, Cockayne's syndrome, Neil-Dingwall syndrome, and progeria. ＋ 2. A method for treating a condition that would benefit from increased levels, comprising orally delivering to a mammal in need of such treatment an effective amount of a compound having formula (VII) as described in claim 1.

17. 17. The method of claim 16, wherein the compound having formula (VII) of claim 1 is crystalline nicotinamide riboside triacetate chloride having Form I.

18. 1. A method for preparing an aqueous solution of crystalline nicotinamide riboside triacetate chloride having Form I, the method comprising providing a crystalline nicotinamide riboside triacetate chloride compound having Form I and contacting the compound with water.

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