Crystalline solids of nicotinic acid mononucleotides and esters thereof and methods of making and using same

Abstract

The present disclosure relates to crystalline solids of nicotinic acid mononucleotides and esters thereof and methods of making and using the same. The present disclosure further relates to crystalline solids comprising a compound of formula (I) wherein R is a n-propyl group, and to a process for making a compound of formula (I) wherein R is a C1-C4 alkyl group or a C2-C4 alkenyl group. The present disclosure also relates to crystalline solids comprising a compound of formula (II), (II) The disclosure further relates to processes for preparing crystalline solids, pharmaceutical formulations of crystalline solids, and the use of such pharmaceutical formulations in the treatment of diseases and conditions.

Description

[0001] This application is a divisional application of Chinese patent application No. 202280037999.X, entitled "Crystal solid of nicotinic acid mononucleotide and its ester and method of preparation and use thereof", which was filed on May 26, 2022, under PCT international application PCT / US2022 / 031124 and entered the Chinese national phase on November 27, 2023.

[0002] Cross-references to related applications This application claims priority to U.S. Provisional Application No. 63 / 193,905, filed May 27, 2021, which is incorporated herein by reference in its entirety for all purposes. Technical Field

[0003] This application relates to crystalline solids of nicotinic acid mononucleotide and its esters, as well as methods for their preparation and use. Background Technology

[0004] Nicotinamide adenine dinucleotide (NAD) and related compounds are known to be essential coenzymes in cellular redox reactions in all living organisms. Extensive evidence also indicates that NAD is involved in many important signaling pathways in mammalian cells, including polymerization (ADP-ribosylation) in DNA repair, mono-ADP-ribosylation in immune responses and G protein-coupled signaling, and the synthesis of cyclic ADP-ribose and nicotinic adenine dinucleotide phosphate (NAADP) in intracellular calcium signaling. NAD and its metabolites have also been shown to play important roles in transcriptional regulation. In particular, the discovery of Sir2 NAD-dependent deacetylase activity has drawn attention to this role of NAD. Despite advances in understanding the biology of NAD, there remains a need for improved compositions and methods for using such compositions for pharmacological intervention and / or manipulation of NAD pathways in living cells and tissues.

[0005] Nicotinic acid mononucleotide (also known as nicotinic acid ribonucleotide) and certain nicotinic acid mononucleotide derivatives are thought to increase cellular NAD production (Sauve, U.S. Patent 10,961,268 B2). However, these compounds are difficult to synthesize on a pharmaceutically appropriate scale with sufficient purity. Given the therapeutic benefits associated with nicotinic acid mononucleotide and its derivatives, there is a need for improved compositions and methods for preparing such compositions. Summary of the Invention

[0006] This disclosure relates to compounds, crystalline solids, and compositions of compounds and / or crystalline solids for regulating nicotinamide adenine dinucleotide (NAD, also known as its oxidized form NAD+ and its reduced form NADH).

[0007] One aspect of this disclosure relates to crystalline solids comprising compounds of formula (I), (I) Where R is n-propyl (compound 1).

[0008] A further aspect of this disclosure relates to crystalline solids comprising compounds of formula (II), (II).

[0009] A further aspect of this disclosure relates to a crystalline solid comprising compound 1: .

[0010] In some embodiments, the crystalline solid has 2θ values ​​of 16.1, 20.1, and 24.5.

[0011] In some embodiments, the solid is anhydrous.

[0012] In some embodiments, the solid is selected from methanol solvates, ethanol solvates, 1-propanol solvates, 2-propanol solvates, C-4 alcohol solvates, C-5 alcohol solvates, and C-6 alcohol solvates, preferably methanol solvates.

[0013] In some embodiments, the crystalline solid contains residual unsolvated solvent or residual nonhydrated water.

[0014] In some embodiments, the crystalline solid contains less than about 5% by weight of propyl nicotinic acid, preferably less than about 1% by weight of propyl nicotinic acid.

[0015] In some embodiments, the crystalline solid comprises less than about 5% by weight of compound 2: .

[0016] In some embodiments, the crystalline solid contains less than about 1% by weight of compound 2.

[0017] In some embodiments, the crystalline solid comprises at least about 90% by weight of compound 1.

[0018] In some embodiments, the crystalline solid comprises at least about 95% by weight of compound 1.

[0019] In some embodiments, the crystalline solid comprises at least about 99% by weight of compound 1.

[0020] In some embodiments, the crystalline solid is non-hygroscopic.

[0021] In some embodiments, the crystalline solid remains stable at a relative humidity of less than about 70%.

[0022] A further aspect of this disclosure relates to a pharmaceutical composition comprising the crystalline solid described in this application and one or more pharmaceutically acceptable excipients.

[0023] A further aspect of this disclosure relates to a method for preparing the crystalline solid described in this application, comprising: a) Dissolving compound 1 in a solvent to form a solution; and b) Crystallize compound 1 from solution to form a crystalline solid.

[0024] In some embodiments, the solvent is methanol.

[0025] In some embodiments, the solution is anhydrous.

[0026] In some embodiments, the temperature of the solvent during the dissolution step is about 30 to about 40°C.

[0027] In some embodiments, compound 1 is completely dissolved in a solvent prior to the crystallization step.

[0028] In some embodiments, crystallization includes forming a supersaturated solution from the solution, wherein the supersaturated solution is supersaturated relative to compound 1.

[0029] In some embodiments, forming a supersaturated solution includes adding an antisolvent to the solution, lowering the temperature of the solution, reducing the volume of the solution, or any combination thereof.

[0030] In some embodiments, forming a supersaturated solution involves lowering the temperature of the solution.

[0031] In some embodiments, forming a supersaturated solution includes lowering the temperature of the solution from about 0°C to about 25°C.

[0032] In some embodiments, forming a supersaturated solution includes adding an antisolvent to the solution.

[0033] In some embodiments, the antisolvent is selected from EtOAc, iPrOAc, TBME, MIBK, THF, 1-propanol, 2-propanol and EtOH, preferably denatured EtOH.

[0034] In some embodiments, the antisolvent is TBME.

[0035] In some embodiments, the solvent to antisolvent ratio is from about 1:1 to about 8:1 by volume.

[0036] In some embodiments, the solvent to antisolvent ratio is approximately 5:1 by volume.

[0037] In some embodiments, the supersaturated solution has a supersaturation ratio of about 1 to 4.

[0038] In some embodiments, the supersaturated solution has a supersaturation ratio of about 2.

[0039] In some embodiments, crystallization includes adding a seed crystal to the solution, wherein the seed crystal comprises compound 1.

[0040] In some embodiments, the method further includes separating the crystalline solid.

[0041] In some embodiments, the method further includes drying the crystalline solid under reduced pressure.

[0042] In some embodiments, the method does not include performing chromatographic analysis to purify compound 1.

[0043] In some embodiments, the method does not include lyophilization during the purification of compound 1.

[0044] In some embodiments, the weight of the crystalline solid is at least about 100 mg.

[0045] In some embodiments, the amount of crystalline solid is at least about 1 g.

[0046] In some embodiments, the crystalline solid contains less than about 1% propyl nicotinate.

[0047] In some embodiments, the crystalline solid comprises at least about 90% of compound 1.

[0048] In some embodiments, the crystalline solid comprises at least about 95% of compound 1.

[0049] In some embodiments, the crystalline solid comprises at least about 99% of compound 1.

[0050] In some embodiments, the crystalline solid is formed according to the method described above.

[0051] A further aspect of this disclosure relates to a crystalline solid comprising a compound of formula (II), (II).

[0052] In some embodiments, the crystalline solid has 2θ values ​​of 21.5, 24.2, 26.7, and 19.6.

[0053] In some embodiments, the compound of formula (II) is a hydrate.

[0054] In some embodiments, the crystalline solid exhibits greater than 90% stability after 14 days of storage at elevated temperatures, as measured by the percentage of area under the curve at 254 nm.

[0055] In some embodiments, the solid is selected from methanol solvates, ethanol solvates, 1-propanol solvates, 2-propanol solvates, C-4 alcohol solvates, C-5 alcohol solvates, and C-6 alcohol solvates.

[0056] In some embodiments, the crystalline solid contains residual unsolvated solvent or residual nonhydrated water.

[0057] In some embodiments, the crystalline solid contains less than about 5% by weight of nicotinic acid, preferably less than about 1% by weight of nicotinic acid.

[0058] In some embodiments, the crystalline solid contains less than about 1% nicotinic acid riboside.

[0059] In some embodiments, the crystalline solid comprises at least about 90% of the compound of formula (II), preferably at least about 95% of the compound of formula (II).

[0060] In some embodiments, the crystalline solid comprises at least about 99% of the compound of formula (II).

[0061] In some embodiments, the average size of a single crystal of the crystalline solid is 20 to 500 micrometers.

[0062] A further aspect of this disclosure relates to a pharmaceutical composition comprising the crystalline solid described in this application and one or more pharmaceutically acceptable excipients.

[0063] A further aspect of this disclosure relates to a method for preparing the crystalline solid described in this application, comprising: a) Dissolving the compound of formula (II) in a solvent to form a solution; and b) Crystallize the compound of formula (II) from solution to form a crystalline solid.

[0064] In some embodiments, the solvent includes water.

[0065] In some embodiments, the solvent includes an alcohol.

[0066] In some embodiments, the alcohol is 1-propanol.

[0067] In some embodiments, the temperature of the solvent during the dissolution step is ambient temperature.

[0068] In some embodiments, the compound of formula (II) is completely dissolved in a solvent prior to the crystallization step.

[0069] In some embodiments, crystallization includes forming a supersaturated solution from the solution, wherein the supersaturated solution is supersaturated relative to the compound of formula (II).

[0070] In some embodiments, forming a supersaturated solution includes adding an antisolvent to the solution, lowering the temperature of the solution, reducing the volume of the solution, or any combination thereof.

[0071] In some embodiments, forming a supersaturated solution involves lowering the temperature of the solution.

[0072] In some embodiments, forming a supersaturated solution includes adding an antisolvent to the solution.

[0073] In some embodiments, the antisolvent is an alcohol.

[0074] In some embodiments, the antisolvent is 1-propanol.

[0075] In some embodiments, the solvent to antisolvent ratio is from about 1:0 to about 1:2 by volume.

[0076] In some embodiments, the ratio of solvent to antisolvent is approximately 6:7.

[0077] In some embodiments, the supersaturated solution has a supersaturation ratio of about 1 to 4.

[0078] In some embodiments, the supersaturated solution has a supersaturation ratio of about 2.

[0079] In some embodiments, forming a supersaturated solution does not involve adding an antisolvent to the solution.

[0080] In some embodiments, the ratio of the compound of formula (II) to the solvent is about 1:3 by weight.

[0081] In some embodiments, crystallization includes adding a seed crystal to the solution, wherein the seed crystal comprises a compound of formula (II).

[0082] In some embodiments, the method further includes separating the crystalline solid.

[0083] In some embodiments, the method further includes drying the crystalline solid under reduced pressure.

[0084] In some embodiments, the method does not include performing chromatographic analysis to purify the compound of formula (II).

[0085] In some embodiments, the method does not include lyophilizing to purify the compound of formula (II).

[0086] In some embodiments, the weight of the crystalline solid is at least about 100 mg.

[0087] In some embodiments, the amount of crystalline solid is at least about 1 g.

[0088] In some embodiments, the crystalline solid contains less than about 1% nicotinic acid riboside.

[0089] In some embodiments, the crystalline solid comprises at least about 90% of the compound of formula (II).

[0090] In some embodiments, the crystalline solid comprises at least about 95% of the compound of formula (II).

[0091] In some embodiments, the crystalline solid comprises at least about 99% of the compound of formula (II).

[0092] In some embodiments, the crystalline solid is formed according to the method described above.

[0093] A further aspect of this disclosure relates to a compound of formula (I), (I) The method, Wherein R is a C1-C6 alkyl or C2-C6 alkenyl; the method includes: in the presence of an acid, causing the compound of formula (II) to... (II), Contact with alcohol R-OH.

[0094] In some embodiments, R is a C1-C6 alkyl group.

[0095] In some embodiments, R is a C3 alkyl group.

[0096] In some embodiments, R is n-propyl.

[0097] In some embodiments, the acid is HCl.

[0098] In some embodiments, the compound of formula (II) is provided as the crystalline solid described in this application.

[0099] In some embodiments, the method further includes preparing a crystalline solid comprising a compound of formula (II) according to any method described in this application.

[0100] In some embodiments, the compound of formula (I) is provided as the crystalline solid described in this application.

[0101] In some embodiments, the method further includes preparing a crystalline solid comprising a compound of formula (I) according to any method described in this application.

[0102] A further aspect of this disclosure relates to a method for increasing NAD levels in a subject, the method comprising administering the crystalline solid described in this application to the subject.

[0103] A further aspect of this disclosure relates to a method for treating or preventing a disease or ailment of a subject, the method comprising administering the crystalline solid described in this application to the subject.

[0104] In some embodiments, a disease or ailment is associated with NAD biosynthesis.

[0105] In some embodiments, the disease or ailment is a neurological or neurodegenerative disorder.

[0106] In some embodiments, the neurological or neurodegenerative disorders are selected from Alzheimer's disease (AD), dementia other than Alzheimer's disease, Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), diffuse Lewy body disease, chorea-acanthocytosis, primary lateral sclerosis, multiple sclerosis (MS), eye diseases, spinal muscular atrophy, chemotherapy-induced neuropathy, diabetes-induced neuropathy, and Friedrich's ataxia.

[0107] In some embodiments, the disease or illness is a symptom caused by infection with COVID-19.

[0108] In some embodiments, the disease or ailment is acute kidney injury (AKI) or chronic kidney disease, such as nephrotic syndrome.

[0109] In some embodiments, the disease or illness is caused by or associated with a cytokine storm.

[0110] In some embodiments, disease or ailment is inflammation.

[0111] In some embodiments, inflammation is selected from multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, degenerative joint disease, spondyloarthritis, gouty arthritis, systemic lupus erythematosus, juvenile arthritis, rheumatoid arthritis, osteoarthritis, osteoporosis, diabetes, menstrual pain, cystic fibrosis, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, mucinous colitis, ulcerative colitis, gastritis, esophagitis, pancreatitis, peritonitis, Alzheimer's disease, shock, ankylosing spondylitis, gastritis, conjunctivitis, pancreatitis, multiple organ injury syndrome, myocardial infarction, atherosclerosis, stroke, reperfusion injury, acute glomerulonephritis, vasculitis, thermal injury, necrotizing enterocolitis, granulocyte transfusion-related syndrome, and Sjögren's syndrome.

[0112] In some embodiments, the disease or ailment is cancer.

[0113] In some embodiments, the cancer is selected from brain cancer, kidney cancer, breast cancer, prostate cancer, testicular cancer, ovarian cancer, lung cancer, colorectal cancer, cervical cancer, skin cancer, stomach cancer, lymphoma, and leukemia.

[0114] In some embodiments, the disease or ailment is a muscle disorder or ailment, such as sarcopenia.

[0115] In some embodiments, the disease or ailment is a solid organ disease or ailment, preferably selected from liver diseases or ailments caused by alcohol-induced cirrhosis.

[0116] In some embodiments, a disease or ailment is treated by transplanting a solid organ, wherein the solid organ is treated with a crystalline solid prior to transplantation.

[0117] In some embodiments, the crystalline solid is administered in unit doses of about 1 to 3000 mg.

[0118] In some embodiments, the crystalline solid is administered in unit doses of about 100 to 1000 mg.

[0119] In some embodiments, the crystalline solid is applied in an amount of about 250 to 750 mg.

[0120] In some embodiments, the crystalline solid is applied twice daily.

[0121] In some embodiments, the subject is a human being.

[0122] A further aspect of this disclosure relates to a method for purifying a compound, the method comprising: a) Dissolving a compound selected from compound 1 or compound 2 in a solvent to form a solution; and b) Crystallize the compound from the solution to form a solid with a purity greater than that of the compound dissolved in step (a).

[0123] In some embodiments, the compound is compound 1.

[0124] In some embodiments, the compound is compound 2.

[0125] In some embodiments, step (a) is performed using an amorphous compound.

[0126] In some embodiments, the higher purity solid in step (b) is greater than about 95% pure.

[0127] In some embodiments, this disclosure relates to methods for preparing such compounds, crystalline solids, and compositions, as well as compounds and compositions of formula (I) wherein R is a C1-C4 alkyl or C2-C4 alkenyl. In some embodiments, this disclosure relates to pharmaceutical compositions comprising one or more NAD-regulating compounds and / or crystalline solids as a first component in combination with one or more active pharmaceutical ingredients. In further embodiments, this disclosure relates to methods for using such compounds, crystalline solids, and / or compositions to promote an increase in intracellular levels of nicotinamide adenine dinucleotide (NAD) in cells and tissues to treat diseases and / or improve cell and tissue survival. Attached Figure Description

[0128] Figure 1A The experimentally obtained XRD pattern of compound 1 as a crystalline solid is shown. Figure 1B The overlay diagram is shown, where the top pattern is the experimental diffraction pattern of compound 1 at room temperature; and the bottom pattern is the calculated diffraction pattern of compound 1 simulated at 100 K. The subtle differences between the simulated and experimental diffraction patterns can be attributed to variations in the lattice with temperature and preferred orientation. Figure 1C It is a representation of the crystal lattice cell of compound 1.

[0129] Figure 2A The experimentally obtained XRD pattern of compound 2 as a crystalline solid is shown. Figure 2B The simulated XRD pattern of compound 2 as a crystalline solid is shown. Figure 2C The overlay diagram is shown, where the top pattern is the experimental diffraction pattern of compound 2; and the bottom pattern is the diffraction pattern calculated from the single-crystal X-ray structure. The subtle differences between the simulated and experimental diffraction patterns can be attributed to variations in the lattice with temperature and preferred orientation.

[0130] Figure 3 The proton NMR spectrum of compound 1, obtained experimentally in DMSO-d6, is shown. The x-axis shows the chemical shift (ppm).

[0131] Figure 4 The proton NMR spectrum of compound 2, obtained experimentally in D2O, is shown. The x-axis shows the chemical shift (ppm). Detailed Implementation

[0132] definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Unless otherwise stated, the following terms as used herein have the meanings assigned to them as follows.

[0133] In this disclosure, terms such as “comprises,” “comprising,” “containing,” and “having” may have the meanings given to them under U.S. Patent Law and may mean “includes,” “including,” etc.; “consisting essentially of” or “consists essentially” also have the meanings given under U.S. Patent Law and the term is open-ended, allowing for more than those listed, provided that the essential or novel features of the listed are not altered by the presence of more than those listed, but excluding prior art embodiments.

[0134] The ranges provided in this document should be understood as shorthand notations of all values ​​within that range. For example, the range 1 to 50 should be understood as any number, combination of numbers, or subrange of numbers including 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

[0135] As used herein, the phrase “a” or “an” refers to one or more of the entities; for example, “compound” refers to one or more compounds or at least one compound. Thus, the terms “a” or “an”, “one or more”, and “at least one” are used interchangeably herein.

[0136] Unless specifically stated or obvious from the context, as used herein, the term “about” is understood to mean within the normal tolerance range in the field, such as within 2 standard deviations of the mean. “About” can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the specified value. Unless the context otherwise requires, all numerical values ​​provided herein are modified by the term “about”.

[0137] As used herein, the term "alkyl" refers to a branched or unbranched saturated hydrocarbon group having 1 to 20 carbon atoms, preferably 1 to 10 carbon atoms. Examples of straight-chain and branched alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, and octyl. Alkyl groups can be cyclic or acyclic. Alkyl groups can be branched or unbranched (i.e., linear). "Lower alkyl" refers to a group containing one to six ( For example Alkyl groups consisting of one to four carbon atoms.

[0138] As used in this article, the term "alkenyl" refers to an aliphatic group containing at least one double bond.

[0139] As used herein, the terms “optional” or “optionally” mean that an event or condition described below may but not necessarily occur, and the description includes examples in which the event or condition occurs as well as examples in which the event or condition does not occur. For example, “optional key” means that the key may or may not exist, and the description includes single, double, or triple keys.

[0140] As described herein, the term "purified" refers to the purity of a given compound. For example, a compound is "purified" when it is the major component of a composition, i.e., when it is at least about 50% w / w pure. Thus, "purified" includes at least about 50% w / w purity, at least about 60% w / w purity, at least about 70% purity, at least about 80% purity, at least about 85% purity, at least about 90% purity, at least about 92% purity, at least about 94% purity, at least about 96% purity, at least about 97% purity, at least about 98% purity, at least about 99% purity, at least about 99.5% purity, and at least about 99.9% purity, wherein "substantially pure" includes at least about 97% purity, at least about 98% purity, at least about 99% purity, at least about 99.5% purity, and at least about 99.9% purity.

[0141] As described in this article, the term "metabolite" refers to compounds that are produced in the body after administration to a subject.

[0142] The term "salt" as used herein refers to a compound comprising both cations and anions, which can be produced by protonation of the proton acceptor portion and / or deprotonation of the proton donor portion. It should be noted that protonation of the proton acceptor portion results in the formation of a cation, in which the charge is balanced by the presence of a physiological anion, while deprotonation of the proton donor portion results in the formation of an anion, in which the charge is balanced by the presence of a physiological cation.

[0143] The phrase “pharmaceutically acceptable salt” refers to a pharmaceutically acceptable salt. Examples of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts formed from inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or formed from organic acids, such as acetic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, dodecyl sulfate, gluconic acid, glutamic acid, salicylic acid, mucoconic acid, etc.; or (2) base addition salts formed from the conjugate base of any of the inorganic acids listed above, wherein the conjugate base contains a component selected from Na+. + K + Mg 2+ Ca 2+ NH g R 4-g + The cationic component, wherein R is C 1-3Alkyl group, and g is a number selected from 0, 1, 2, 3, or 4. It should be understood that all pharmaceutically acceptable salts mentioned include solvation forms (solvents) or crystalline solids of the same acid addition salts as defined herein.

[0144] This disclosure also includes useful forms of the compounds disclosed herein, such as metabolites, solvates, prodrugs, salts, especially pharmaceutically acceptable salts, and / or coprecipitates.

[0145] The compounds disclosed herein can exist as solvates, wherein the compounds form crystals containing molecules of a polar solvent such as water, methanol, or ethanol as structural elements of the compound's crystal lattice. The molecules of the polar solvent can exist with the molecules of the compound in stoichiometric or non-stoichiometric ratios. In the case of stoichiometric solvates, For example Half-, (half-), single-, double-half-, two-, three-, four-, five- wait The solvates are all possible. This disclosure includes all such solvates.

[0146] Furthermore, the compounds disclosed herein may exist in a free form. For example It may exist as a free base, or as a free acid, or as a zwitterion, or in the form of a salt. The salt may be any salt, organic or inorganic addition salt, particularly any pharmaceutically acceptable organic or inorganic addition salt that is commonly used in pharmaceutical manufacturing or for, for example, the isolation or purification of the compounds disclosed herein.

[0147] The term "subject" intended for use includes, but is not limited to, humans (i.e., men or women of any age group, such as pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young adults, middle-aged adults, or older adults)) and / or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and / or dogs; and / or birds, including commercially relevant birds such as chickens, ducks, geese, quails, and / or turkeys.

[0148] The terms “treatment,” “treating,” “relief,” and “improvement” are used interchangeably herein. These terms refer to a method of achieving a beneficial or desired outcome, including but not limited to therapeutic and / or preventative benefits. A therapeutic benefit means the eradication or relief of an underlying condition that is being treated. Furthermore, a therapeutic benefit is achieved by eradicating or relieving one or more physical symptoms associated with an underlying condition, resulting in an improvement observed in the patient, although the patient may still suffer from the underlying condition. For preventative benefits, pharmaceutical compounds and / or compositions may also be administered to patients at risk of developing a specific disease or to patients reporting one or more physical symptoms of a disease, even if the disease has not yet been diagnosed.

[0149] As used herein, “prevention” of a disease or condition refers to a compound and / or its crystalline solid that reduces the occurrence of a disease or condition in a treated sample relative to an untreated control sample, or that delays the onset of one or more symptoms of a disease or condition relative to an untreated control sample and reduces its severity.

[0150] The term "treatment" includes preventative and / or therapeutic treatment. The term "preventative or therapeutic" treatment is recognized in the art and includes administering one or more of the disclosed compositions to a subject. Treatment is preventative (i.e., it protects the subject from developing the unwanted condition) if administered before the clinical manifestation of an unwanted condition (e.g., a disease or other unwanted state of the subject), and therapeutic (i.e., it aims to reduce, improve, or stabilize the existing unwanted condition or its side effects) if administered after the manifestation of the unwanted condition.

[0151] The terms “formulation” or “dosage form” are intended to include solid and liquid formulations of an active compound and / or its crystalline solid, and those skilled in the art will understand that the active ingredient may be present in different formulations depending on the required dosage and pharmacokinetic parameters.

[0152] As used herein, the term "excipient" refers to a compound used in the preparation of a pharmaceutical composition and is generally safe, non-toxic, neither biologically nor otherwise required, and includes excipients acceptable for veterinary and human pharmaceutical use.

[0153] As used herein, the phrase “pharmaceutically acceptable” refers to compounds, materials, compositions, and / or dosage forms that, within reasonable medical judgment, are suitable for contact with a subject’s tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and that are commensurate with a reasonable benefit / risk ratio.

[0154] As used herein, the phrase “pharmaceutically acceptable carrier” means a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation and harmless to the subject.

[0155] As used herein, the phrase "combined administration" refers to any form of administration of two or more different therapeutic agents such that the second agent is administered while the previously administered agent is still effective in the body (e.g., both agents are effective simultaneously in the patient, which may include the synergistic effect of the two agents). For example, different therapeutic compounds may be administered simultaneously or sequentially in the same formulation or in separate formulations. Thus, an individual receiving such treatment may benefit from the combined effect of the different therapeutic agents.

[0156] The description of a list of elements in any definition of a variable herein includes defining that variable as any single element or combination (or sub-combination) of the listed elements. The description of embodiments herein includes embodiments as any single embodiment, or embodiments combined with any other embodiment or part thereof.

[0157] It should be understood that whatever values ​​and ranges are provided herein, all values ​​and ranges encompassed by these values ​​and ranges are included within the scope of this disclosure. Furthermore, all values ​​falling within these ranges, as well as the upper or lower limits of the value ranges, are also contemplated in this application.

[0158] Compounds and crystalline solids In one aspect, this disclosure provides crystalline solids comprising compounds of formula (I), (I) Where R is n-propyl. Compounds of formula (I) where R is n-propyl are alternatively referred to herein as "compound 1".

[0159] In some embodiments, the crystalline solids described herein are characterized by X-ray diffraction (XRD). In some embodiments, XRD is X-ray powder diffraction (XRPD). θ represents the diffraction angle, measured in degrees. In some embodiments, the diffractometer used in XRD measures a diffraction angle that is twice the diffraction angle θ. Therefore, in some embodiments, the diffraction pattern described herein refers to the X-ray intensity measured at an angle 2θ.

[0160] In some embodiments, the crystalline solid containing the compound of formula (I) has 2θ values ​​of 16.1, 20.1, and 24.5. In some embodiments, the crystalline solid containing the compound of formula (I) has 2θ values ​​of 16.1, 20.1, 24.5, 23.7, 18.8, 21.5, 17.7, 8.1, 9.9, 13.0, 26.3, and 30.4. In some embodiments, the crystalline solid containing the compound of formula (I) has 2θ values ​​of 16.1, 20.1, 24.5, 23.7, 18.8, 21.5, 17.7, 8.1, 9.9, 13.0, 26.3, 30.4, 23.0, 26.6, 25.3, 25.5, and 19.7. In some embodiments, the crystalline solid containing the compound of formula (I) has an XRD pattern substantially as shown in FIG1 (A or B). In some embodiments, the XRD pattern is a methanol solvate of the compound of formula (I). In some embodiments, the XRD pattern is a non-solvate of the compound of formula (I). In some embodiments, the XRD pattern corresponds to... Figure 1C The corresponding three-dimensional shape.

[0161] In some embodiments, the compound of formula (I) is not solvated or hydrated in the crystalline solid (e.g., the lattice does not contain molecules of solvent or water). In some embodiments, the crystalline solid containing the compound of formula (I) contains unhydrated water and / or unsolvated solvent. In some embodiments, such unhydrated water and / or unsolvated solvent are present in residual amounts (e.g., less than 10% by weight, or less than 5% by weight), or in amounts greater than zero but less than 1% by weight.

[0162] In some embodiments, the compound of formula (I) is solvated by one or more solvents. In some embodiments, the compound of formula (I) is solvated by an alcohol to form an alcohol solvate, preferably by methanol to form a methanol solvate. In some embodiments, the crystalline methanol solvate of the compound of formula (I) contains about 1.0, about 1.1, or about 1.2 molecules of methanol relative to one molecule of the compound of formula (I). In some embodiments, the compound of formula (I) is solvated by ethanol. In some embodiments, the compound of formula (I) is solvated by water. In some embodiments, the compound of formula (I) is solvated / hydrated by ethanol and water. In various embodiments, the crystalline solid is a solvate selected from methanol solvates, ethanol solvates, 1-propanol solvates, 2-propanol solvates, C-4 alcohol solvates, C-5 alcohol solvates, and C-6 alcohol solvates, preferably methanol solvates.

[0163] In various embodiments, compounds of formula (I) having the XRD patterns disclosed herein are prepared from amorphous materials with a purity greater than 90%, including the steps of dissolving the amorphous material in an alcohol and allowing the product to precipitate over time, preferably at ambient temperature. In various embodiments, compounds of formula (I) are prepared from amorphous materials by a method comprising the steps of dissolving the amorphous material in water or an aqueous solution, then diluting the resulting solution with an antisolvent, and allowing the compound of formula (I) to precipitate over time. In various embodiments, the antisolvent is an alcohol, such as ethanol, methanol, propanol, or another alcohol having eight or fewer carbon atoms. In various embodiments, the antisolvent is ethanol. In various embodiments, the antisolvent is denatured ethanol.

[0164] In one aspect, this disclosure provides crystalline solids comprising compounds of formula (II), (II).

[0165] The compound of formula (II) is referred to herein as “compound 2”.

[0166] In some embodiments, the crystalline solid containing the compound of formula (II) has 2θ values ​​of 21.5, 24.2, 26.7, and 19.6. In some embodiments, the crystalline solid containing the compound of formula (II) has 2θ values ​​of 21.5, 24.2, 26.7, 19.6, 15.6, and 29.3. In some embodiments, the crystalline solid containing the compound of formula (II) has 2θ values ​​of 21.5, 24.2, 26.7, 19.6, 15.6, 29.3, 22.9, 23.1, 22.5, 13.3, 22.2, 30.0, 30.6, 13.1, 27.2, and 17.5. In some embodiments, the crystalline solid containing the compound of formula (II) has an XRD pattern substantially as shown in FIG2. In some embodiments, the XRD pattern is a hydrate of the compound of formula (II).

[0167] In some embodiments, the compound of formula (II) is not solvated or hydrated in the crystalline solid (e.g., the lattice does not contain molecules of solvent or water). In some embodiments, the compound of formula (II) is solvated by one or more solvents. In some embodiments, the crystalline solid containing the compound of formula (II) contains non-hydrated water and / or non-solventizing solvent. In some embodiments, such non-hydrated water and / or non-solventizing solvent are present in residual amounts (e.g., less than 10% by weight, or less than 5% by weight), or in amounts greater than zero but less than 1% by weight.

[0168] In some embodiments, the compound of formula (II) is solvated with water to form a hydrate. In other embodiments, the compound of formula (II) is solvated with an alcohol to form an alcohol solvate. In some embodiments, the crystalline hydrate of the compound of formula (II) contains about 1.0, about 1.1, or about 1.2 molecules of methanol relative to one molecule of the compound of formula (II). In some embodiments, the compound of formula (II) is solvated with ethanol. In some embodiments, the compound of formula (II) is solvated / hydrated with both ethanol and water. In various embodiments, the crystalline solid comprising the compound of formula (II) is a solvate selected from methanol solvates, ethanol solvates, 1-propanol solvates, 2-propanol solvates, C-4 alcohol solvates, C-5 alcohol solvates, and C-6 alcohol solvates, preferably methanol solvates.

[0169] In various embodiments, compounds of formula (II) having the XRD patterns disclosed herein are prepared from amorphous materials with a purity greater than 90% by a method comprising the steps of: dissolving the amorphous material in an alcohol, and precipitating the compound of formula (II) over time, preferably at ambient temperature. In various embodiments, compounds of formula (II) are prepared from amorphous materials by a method comprising the steps of: dissolving the amorphous material in water or an aqueous solution, then diluting the resulting solution with an antisolvent, and precipitating the compound of formula (II) over time. In various embodiments, the antisolvent is an alcohol, such as ethanol, methanol, propanol, or another alcohol having eight or fewer carbon atoms. In various embodiments, the antisolvent is ethanol. In various embodiments, the antisolvent is denatured ethanol.

[0170] It is evident that the compounds of formula (I) and (II) can exist in a variety of protonated states, depending in particular on the pH of their environment. At various pH environments, the compounds of formula (I) and (II) exist as zwitterions or internal salts, as illustrated herein.

[0171] In various embodiments, the compounds of formulas (I) and (II) are one or more salts, wherein the salts are selected from H... + Li + Na + K + Mg 2+ and Ca 2+ The salt is formed from cations selected from acetate, trifluoromethansulfonate (triflate), halides, trifluoroacetate, formate, and H2PO4. - HPO4 2- OH - HSO4- SO4 2- NO3 - HCO3 - and CO3 2- Anions are formed from compounds of [specific compounds] and their mixtures. In various embodiments, the compounds are zwitterions.

[0172] This disclosure includes the use of pharmaceutically acceptable salts and / or crystalline solids of the compounds disclosed herein. In some embodiments, the intended salts of this disclosure include, but are not limited to, alkyl, dialkyl, trialkyl, or tetraalkylammonium salts. In some embodiments, the intended salts of the present invention include, but are not limited to, L-arginine, phenethylbenzylamine, benzathine penicillin, betaine, calcium hydroxide, choline, dimethylethanolamine, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucosamine, hepaticillin, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts.

[0173] In some embodiments, the compound is a salt having an anion selected from acetate, trifluoromethanesulfonate, halide, trifluoroacetate, or formate. In other embodiments, if the disclosed compound is in contact with a medium (e.g., an aqueous medium), the anion may be selected from, for example, OH. - H2PO4 - HPO4 2- HSO4 - SO4 2- NO3 - HCO3 - and CO3 2- .

[0174] In some embodiments, the disclosed compound is in the form of a negatively charged phosphate that can form a salt with any suitable cation. The cation can be changed when the compound is isolated or transferred to a medium with different anionic substances. For example, the disclosed compound can be in the form of a phosphate that is a pharmaceutically acceptable salt as described herein. In some embodiments, the cation may be selected from Li. + Na + K + Mg 2+ and Ca 2+ .

[0175] In some embodiments, the crystalline solids described herein are not part of a solution, suspension, mixture, slurry, reaction mixture, or the like.

[0176] In some embodiments, the average size of a single crystal of a crystalline solid comprising a compound of formula (I) or (II) is greater than about 1 micrometer, greater than about 5 micrometers, greater than about 10 micrometers, or greater than about 20 micrometers. In further embodiments, the average size of a single crystal of a crystalline solid comprising a compound of formula (I) or (II) is about 1 to about 100 micrometers, about 20 to about 100 micrometers, about 1 to about 500 micrometers, about 1 to about 250 micrometers, about 20 to about 250 micrometers, or about 20 to about 500 micrometers.

[0177] In some embodiments, the crystalline solids described herein have lower solubility in water compared to the amorphous solids of the compounds of formulas (I) and (II). In a preferred embodiment, the solubility ratio of the crystalline solid to water is from about 1:5 to about 1:75 by weight. In a more preferred embodiment, the solubility ratio of the crystalline solid to water is from about 1:10 to about 1:60 by weight. This lower solubility in water can impart desirable therapeutic properties.

[0178] In various embodiments, the crystalline solid is anhydrous. In various embodiments, the crystalline solid contains less than about 5% water, less than about 2% water, less than about 1% water, less than about 0.5% water, or less than about 0.1% water. In some embodiments, the percentages are by weight.

[0179] In preferred embodiments, the compound of formula (I) or its crystalline solid contains less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. For example, in preferred embodiments, the compound of formula (I) or its crystalline solid contains less than about 5% propyl nicotinate, less than about 2% propyl nicotinate, less than about 1% propyl nicotinate, less than about 0.5% propyl nicotinate, or less than about 0.1% propyl nicotinate. In some embodiments, the percentages are by weight.

[0180] In preferred embodiments, the compound of formula (II) or its crystalline solid contains less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. For example, in preferred embodiments, the compound of formula (II) or its crystalline solid contains less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, less than about 0.1% nicotinic acid riboside, or less than about 0.01% nicotinic acid riboside. In some embodiments, the percentages are by weight.

[0181] In some preferred embodiments, the crystalline solid comprising the compound of formula (I) or (II) is pure or substantially pure. In some preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably greater than about 98% pure, for example, greater than about 99% pure. In some embodiments, the percentage is by weight. In preferred embodiments, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of the compound of formula (I) or (II). In some embodiments, the percentage is by weight.

[0182] The crystalline solids described herein may have advantageous properties compared to the amorphous forms of the compounds of formulas (I) and (II). In some embodiments, such as at elevated temperatures, the crystalline solids exhibit improved chemical and / or physical stability. In some embodiments, compositions comprising the crystalline solids exhibit improved chemical and / or physical stability. In some embodiments, the crystalline solids have improved storage stability. In some embodiments, the crystalline solids exhibit better handling properties during manufacturing than the amorphous forms, which may result in higher purity, stability, and / or consistency of the compounds, crystalline solids, and compositions. In some embodiments, the crystalline solids may be easier to process under typical pharmaceutical processing conditions. In some embodiments, improved handling properties include improved viscosity and flow properties. In some embodiments, the crystalline solids described herein are less hygroscopic than the amorphous forms. For example, when exposed to a humid environment (e.g., at least 50% humidity), the crystalline solids may absorb less water than the corresponding amorphous forms under the same conditions. In some embodiments, the crystalline solids maintain structural integrity when exposed to humidity, for example, they may be less prone to swelling or transformation into less stable forms. In some embodiments, the crystalline solids described herein have lower solubility and / or dissolution rates compared to the amorphous forms. In some embodiments, to prolong the effect of the crystalline solid as a drug, it is desirable to slow the absorption of the crystalline solid. For example, in some embodiments, the crystalline solid described herein is efficiently delivered to the intestine and does not dissolve significantly in the stomach. In some embodiments, for example, a prolonged release effect is achieved by using an aqueous suspension of the crystalline solid. In other embodiments, delayed release is achieved by dissolving or suspending the solid material in an oil-based medium. In some embodiments, the crystalline solid described herein has higher purity than its amorphous form and / or is advantageous for the large-scale preparation of pure materials, for example, at a lower cost or using less material or space-intensive purification methods. In some such embodiments, the crystalline solid and methods described herein are advantageous for large-scale purification, for example, greater than about 1 gram, greater than about 10 grams, or greater than about 100 grams.

[0183] Methods for preparing crystalline solids This document also provides a method for preparing a crystalline solid of a compound of formula (I). In some embodiments, this disclosure relates to a method for preparing a crystalline solid of a compound of formula (I), comprising a) dissolving a compound of formula (I) in a solvent to form a mixture; and b) crystallizing a compound of formula (I) from the mixture to form a crystalline solid.

[0184] In a preferred embodiment, the mixture comprising the compound of formula (I) is a solution. In other embodiments, the mixture is a slurry or suspension. In some embodiments, the solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids with similar polarity and properties, and combinations thereof. In some embodiments, the solvent includes acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), methanol, ethanol, ethyl acetate, isopropyl acetate, methanol, methyl ethyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, propanol, butanol, water, or any combination thereof. In some embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including its branched and unbranched isomers. In a preferred embodiment, the solvent is methanol. In some embodiments, the solvent includes two or more of the solvents described herein. In some embodiments, the solution is anhydrous.

[0185] In some embodiments, the solvent temperature is higher than ambient temperature during the dissolution step. In such embodiments, the method includes heating the solvent. For example, the solvent temperature may be about 30 to about 50°C or about 30 to about 40°C, such as about 35°C. In other embodiments, the solvent temperature is about ambient temperature during the dissolution step. In still other embodiments, the solvent temperature is lower than ambient temperature during the dissolution step. In such embodiments, the method includes cooling the solvent. In some embodiments, the solvent temperature is about 20 to about 30°C, such as about 25°C. In a preferred embodiment, the compound of formula (I) is completely dissolved in the solvent prior to the crystallization step. Complete dissolution means that the compound is present in a homogeneous solution, rather than in a slurry or suspension. In other embodiments, the compound of formula (I) is partially dissolved in the solvent prior to the crystallization step.

[0186] In some embodiments, the method includes forming a supersaturated solution from a mixture (e.g., a solution) of compounds of formula (I), wherein the supersaturated solution is supersaturated relative to the compounds of formula (I). In some embodiments, the supersaturated solution has a supersaturation ratio of about 1 to about 4, for example, about 2. In some such embodiments, the compounds of formula (I) are precipitated (e.g., crystallized) from the supersaturated solution. In some embodiments, the resulting precipitate (e.g., crystals) is a crystalline solid as described herein.

[0187] Supersaturated solutions can be formed using a variety of methods. In some embodiments, forming a supersaturated solution may include adding an antisolvent to a mixture (e.g., a solution), lowering the temperature of the mixture (e.g., a solution), reducing the volume of the mixture (e.g., a solution), or any combination thereof. For example, the method may include adding an antisolvent, subsequently cooling the resulting mixture, and then adding additional antisolvent.

[0188] In some embodiments, forming a supersaturated solution involves lowering the temperature of the mixture containing the compound of formula (I). In some such embodiments, the temperature of the solution is lowered to about 0 to about 25°C, about 0 to about 10°C, or about -5 to about 5°C, for example, about 0°C. In some embodiments, cooling the solution can be passive (e.g., allowing the solution to stand at ambient temperature) or active (e.g., cooling the solution in an ice bath or refrigerator).

[0189] In some embodiments, forming a supersaturated solution includes adding an antisolvent to a mixture containing compounds of formula (I). As used herein, "antisolvent" means a liquid in which compounds of formulas (I) and (II) are insoluble, minimally soluble, or partially soluble. In practice, adding an antisolvent to a solution containing compounds of formulas (I) and (II) reduces the solubility of compounds of formulas (I) and (II) in the solvent, thereby promoting precipitation.

[0190] In some embodiments, the antisolvent may be added slowly to prevent uncontrolled crystallization. In some embodiments, the antisolvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids miscible with the solvent of similar polarity and properties, and combinations thereof. In some embodiments, the antisolvent is an alkane solvent, such as hexane or pentane, or an aromatic hydrocarbon solvent, such as benzene, toluene, or xylene. In some embodiments, the antisolvent is selected from ethyl acetate, isopropyl acetate, methyl tert-butyl ether, methyl isobutyl ketone, tetrahydrofuran, 1-propanol, 2-propanol, ethanol, denatured ethanol, and combinations thereof. In a preferred embodiment, the antisolvent is TBME. In some embodiments, the antisolvent comprises two or more of the solvents described herein. In some embodiments, the solvent to antisolvent ratio is from about 1:1 to about 8:1 by volume, or from about 4:1 to about 6:1 by volume, for example, about 5:1 by volume.

[0191] In some embodiments, the method further includes evaporating the solvent from the mixture. In some embodiments, the solvent may be removed under reduced pressure and / or by heating the solvent to evaporate it.

[0192] In some embodiments, crystallization includes inducing secondary nucleation. In some embodiments, crystallization includes adding a seed crystal to the solution, wherein the seed crystal comprises a compound of formula (I). In some embodiments, the seed crystal is formed during a previous crystallization. In some such embodiments, the previous crystallization is carried out on a smaller scale than the crystallization following the addition of the seed crystal.

[0193] In other embodiments, secondary nucleation can be caused by other changes in the mixture environment. For example, crystallization can be promoted by environmental changes, including but not limited to crystallizer walls, stirring impellers, and ultrasonic treatment.

[0194] In a preferred embodiment, the method includes separating the crystalline solid, for example by filtering the crystal, by decanting fluid from the crystal, or by any other suitable separation technique.

[0195] In some embodiments, the method includes washing a crystalline solid containing a compound of formula (II), for example, with a solvent described herein or a mixture of one or more of these solvents and / or antisolvents. In some embodiments, washing the crystalline solid includes washing with a liquid selected from antisolvents, solvents, alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids of similar polarity and properties, and combinations thereof. In some embodiments, the liquid is selected from acetonitrile; N,N-dimethylacetamide (DMA); dimethylformamide (DMF); dimethyl sulfoxide (DMSO); ethyl acetate; isopropyl acetate; methyl ethyl ketone; methyl isobutyl ketone; N-methyl-2-pyrrolidone (NMP); tetrahydrofuran; alcohols such as methanol, ethanol, propanol, or butanol; water; alkane solvents such as pentane, hexane, or heptane; aromatic hydrocarbon solvents such as benzene, toluene, or xylene; methyl tert-butyl ether; and combinations thereof. In some embodiments, the solvent and / or antisolvent are cooled prior to washing. In some embodiments, the method includes drying the crystalline solid, for example under reduced pressure and / or by heating the crystalline solid, and / or under a drying gas such as nitrogen, argon or air.

[0196] In some embodiments, the method for preparing crystalline solids removes one or more impurities from the compound of formula (I). In some embodiments, the method does not include chromatographic analysis or lyophilization to purify the compound of formula (I). In some such embodiments, the methods described herein are used to purify the compound of formula (I), for example, as a final purification step in the manufacture of the compound of formula (I).

[0197] The methods described herein can provide benefits such as removing impurities from compounds of formula (I). In preferred embodiments, the crystalline solid contains less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. In some preferred embodiments, the crystalline solid contains less than about 5% propyl nicotinate, less than about 2% propyl nicotinate, less than about 1% propyl nicotinate, less than about 0.5% propyl nicotinate, or less than about 0.1% propyl nicotinate. In some embodiments, the percentages are by weight.

[0198] In some preferred embodiments, the crystalline solid comprising the compound of formula (II) is pure or substantially pure. In some preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably greater than about 98% pure. In some embodiments, the percentages are by weight.

[0199] In a preferred embodiment, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of a compound of formula (I). In some embodiments, the percentages are by weight.

[0200] Another aspect of this disclosure provides a method for preparing a crystalline solid of a compound of formula (II). In some embodiments, this disclosure relates to a method for preparing a crystalline solid of a compound of formula (II), comprising a) dissolving a compound of formula (II) in a solvent to form a mixture; and b) crystallizing the compound of formula (II) from the mixture to form a crystalline solid. In some embodiments, the method includes reacting nicotinic riboside with a phosphorus-containing group such as phosphoryl chloride prior to the dissolution and crystallization steps to provide a compound of formula (II).

[0201] In a preferred embodiment, the mixture comprising the compound of formula (II) is a solution. In other embodiments, the mixture is a slurry or suspension. In some embodiments, the solvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids with similar polarity and properties, and combinations thereof. In some embodiments, the solvent includes acetonitrile, N,N-dimethylacetamide (DMA), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), methanol, ethanol, ethyl acetate, isopropyl acetate, methanol, methyl ethyl ketone, N-methyl-2-pyrrolidone (NMP), tetrahydrofuran, propanol, butanol, water, or any combination thereof. In some embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including its branched and unbranched isomers. In a preferred embodiment, the solvent is water. In some embodiments, the solvent includes an alcohol, such as 1-propanol. In some embodiments, the solvent includes two or more of the solvents described herein. In some embodiments, the ratio of the compound of formula (II) to the solvent is from about 1:2 to about 1:4 by weight, for example, about 1:3 by weight.

[0202] In some embodiments, the solvent temperature is above ambient temperature during the dissolution step. In such embodiments, the method includes heating the solvent. For example, the solvent temperature may be about 30 to about 50°C or about 30 to about 40°C, such as about 35°C. In other embodiments, the solvent temperature is about ambient temperature during the dissolution step. In still other embodiments, the solvent temperature is below ambient temperature during the dissolution step. In such embodiments, the method includes cooling the solvent. In some embodiments, the solvent temperature is about 20 to about 30°C, such as about 25°C. In a preferred embodiment, the compound of formula (II) is completely dissolved in the solvent prior to the crystallization step. Complete dissolution means that the compound is present in a homogeneous solution, rather than in a slurry or suspension. In other embodiments, the compound of formula (II) is partially dissolved in the solvent prior to the crystallization step.

[0203] In some embodiments, the method includes forming a supersaturated solution from a mixture (e.g., a solution) of compounds of formula (II), wherein the supersaturated solution is supersaturated relative to the compounds of formula (II). In some embodiments, the supersaturated solution has a supersaturation ratio of 1 to about 4, for example, about 2. In some such embodiments, the compounds of formula (I) are precipitated (e.g., crystallized) from the supersaturated solution. In some embodiments, the resulting precipitate (e.g., crystals) is a crystalline solid as described herein.

[0204] Supersaturated solutions can be formed using a variety of methods. In some embodiments, forming a supersaturated solution may include adding an antisolvent to a mixture (e.g., a solution), lowering the temperature of the mixture (e.g., a solution), reducing the volume of the mixture (e.g., a solution), or any combination thereof. For example, the method may include adding an antisolvent, subsequently cooling the resulting mixture, and then adding additional antisolvent.

[0205] In some embodiments, forming a supersaturated solution involves lowering the temperature of the mixture containing the compound of formula (II). In some such embodiments, the temperature of the solution is lowered to about 0 to about 25°C, about 0 to about 10°C, or about -5 to about 5°C, for example, about 0°C. In some embodiments, cooling the solution can be passive (e.g., allowing the solution to stand at ambient temperature) or active (e.g., cooling the solution in an ice bath or refrigerator).

[0206] In some embodiments, forming a supersaturated solution comprises adding an antisolvent to a mixture comprising a compound of formula (II). In some embodiments, the antisolvent may be added slowly to prevent uncontrolled crystallization. In some embodiments, the antisolvent is selected from alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids miscible with the solvent of similar polarity and properties, and combinations thereof. In some embodiments, the antisolvent is an alkane solvent, such as hexane or pentane, or an aromatic hydrocarbon solvent, such as benzene, toluene, or xylene. In some embodiments, the antisolvent is selected from ethyl acetate, isopropyl acetate, methyl tert-butyl ether, methyl isobutyl ketone, tetrahydrofuran, and combinations thereof. In other embodiments, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including its branched and unbranched isomers. In a preferred embodiment, the antisolvent is 1-propanol. In some embodiments, the antisolvent comprises two or more of the solvents described herein. In some embodiments, the solvent to antisolvent ratio is from about 1:0 to about 1:2 by volume, or from about 1:0 to about 6:7 by volume, for example, about 6:7 by volume. In some embodiments, a supersaturated solution is formed without the addition of an antisolvent.

[0207] In some embodiments, the method further includes evaporating the solvent from the mixture. In some embodiments, the solvent may be removed under reduced pressure and / or by heating the solvent to evaporate it.

[0208] In some embodiments, the method further includes adding an acid or base to adjust the pH of the mixture (e.g., a solution) to alter the protonated state of the compound of formula (II). In some embodiments, the base is an amine, such as triethylamine. In some embodiments, the method further includes adding a base to the solution. In some embodiments, the pH of the solution is adjusted to about 2 to about 4, for example, about 3.

[0209] In some embodiments, crystallization includes inducing secondary nucleation. In some embodiments, crystallization includes adding a seed crystal to the solution, wherein the seed crystal comprises a compound of formula (II). In some embodiments, the seed crystal is formed during a previous crystallization. In some such embodiments, the previous crystallization is carried out on a smaller scale than the crystallization following the addition of the seed crystal.

[0210] In other embodiments, secondary nucleation can be caused by other changes in the mixture environment. For example, crystallization can be promoted by environmental changes, including but not limited to crystallizer walls, stirring impellers, and ultrasonic treatment.

[0211] In a preferred embodiment, the method includes separating the crystalline solid, for example by filtering the crystal, by decanting fluid from the crystal, or by any other suitable separation technique.

[0212] In some embodiments, the method includes washing the crystalline solid containing the compound of formula (II), for example, washing the crystalline solid with one or more solvents described herein or mixtures of such solvents and / or antisolvents. In some embodiments, washing the crystalline solid includes washing with a liquid selected from antisolvents, solvents, alcohols, ketones, carboxylic acids, esters, ethers, alkanes, water, amines, other liquids with similar polarity and properties, and combinations thereof. In some embodiments, the liquid is selected from acetonitrile; N,N-dimethylacetamide (DMA); dimethylformamide (DMF); dimethyl sulfoxide (DMSO); ethyl acetate; isopropyl acetate; methyl ethyl ketone; methyl isobutyl ketone; N-methyl-2-pyrrolidone (NMP); tetrahydrofuran; alcohols such as methanol, ethanol, propanol, or butanol; water; alkane solvents such as pentane, hexane, or heptane; aromatic hydrocarbon solvents such as benzene, toluene, or xylene; methyl tert-butyl ether; and combinations thereof. In a preferred embodiment, the method includes washing the crystalline solid with a 2:1 volumetric mixture of 1-propanol and water, optionally followed by washing the crystalline solid with MTBE. In some embodiments, the solvent and / or antisolvent is cooled prior to washing. In some embodiments, the method includes drying the crystalline solid, for example under reduced pressure and / or by heating the crystalline solid.

[0213] In some embodiments, the method for preparing crystalline solids removes one or more impurities from compounds of formula (II). In some embodiments, the method does not include chromatographic analysis or lyophilization to purify compounds of formula (II). In some such embodiments, the methods described herein are used to purify compounds of formula (II), for example, as a final purification step in the manufacture of compounds of formula (II).

[0214] The methods described herein provide benefits such as removing impurities from compounds of formula (II). In preferred embodiments, the crystalline solid contains less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. In some embodiments, the crystalline solid contains less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, or less than about 0.1% nicotinic acid riboside. In some embodiments, percentages are by weight.

[0215] In some preferred embodiments, the crystalline solid comprising the compound of formula (II) is pure or substantially pure. In some preferred embodiments, the crystalline solid is greater than about 90% pure. More preferably, the crystalline solid is greater than about 95% pure, or even more preferably greater than about 98% pure. In some embodiments, the percentages are by weight.

[0216] In a preferred embodiment, the crystalline solid comprises at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, at least about 99.5%, or at least about 99.9% of a compound of formula (II). In some embodiments, the percentages are by weight.

[0217] synthesis In various embodiments, this disclosure provides methods for forming compounds of formula (I). (I) Wherein R is a C1-C6 alkyl or C2-C6 alkenyl; the method comprises: in the presence of an acid, causing the compound of formula (II) to... Contact with alcohol R-OH. See Scheme 1 for example. Option 1 In some embodiments, R is a C1-C6 alkyl group. In some embodiments, R is a C1-C4 alkyl group or a C2-C4 alkenyl group. In some embodiments, R is a C3 alkyl group. In some embodiments, R is n-propyl.

[0218] In some embodiments, the acid is a strong acid. In some embodiments, the acid is an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc. In other embodiments, the acid is an organic acid, such as glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, lauryl sulfate, gluconic acid, glutamic acid, salicylic acid, mucoconic acid, etc. In a preferred embodiment, the acid is HCl.

[0219] In some embodiments, the method includes purifying the compound of formula (II). In some embodiments, the method includes removing an ammonium salt, such as triethylammonium salt, from the compound of formula (II). In some embodiments, the compound of formula (II) is provided as a crystalline solid. In some embodiments, the method includes crystallizing the compound of formula (II) according to the method described herein.

[0220] In some embodiments, the method includes adding a solvent to a compound of formula (II) to form a mixture, such as a solution. In some embodiments, the solvent is a polar solvent. Polar solvents include polar groups, which may be selected from, for example, hydroxyl, carbonyl, ether, ester, amine, amide, and carboxyl groups. In some embodiments, the solvent comprises water. In a preferred embodiment, the alcohol R-OH is the reaction solvent. In a preferred embodiment, the solvent is a straight-chain or branched alcohol, such as methanol, ethanol, propanol, or butanol, including its branched and unbranched isomers. In a more preferred embodiment, the solvent is propanol, such as 1-propanol or 2-propanol. In some embodiments, the temperature of the mixture comprising the compound of formula (II) is about -5 to about 10°C, about -5 to about 5°C, or about 0°C.

[0221] In some embodiments, the method includes mixing a compound of formula (II) with an alcohol. In some embodiments, the mixing step is performed for about 12 to about 72 hours, about 12 to about 48 hours, or about 12 to about 24 hours. In some embodiments, throughout the mixing step, the mixture containing the compound of formula (II) is at about -5 to about 10°C, about -5 to about 5°C, or about 0°C.

[0222] In some embodiments, the method includes adding a base. In various embodiments, the base is added after about 12 to about 72 hours, about 12 to about 48 hours, or about 12 to about 24 hours. In some embodiments, the base is added until the pH of the reaction mixture is about 4 to about 5. In some embodiments, the base is an amine base. In some such embodiments, the base is a trialkylamine base. In a preferred embodiment, the base is triethylamine. In a further embodiment, the method includes adding seed crystals of a compound of formula (I) to the reaction mixture.

[0223] In various embodiments, the method includes purifying the resulting product (i.e., the compound of formula (I)). In some embodiments, the purified product includes chromatographic analysis. In other embodiments, the purified product does not include chromatographic analysis. In a preferred embodiment, the purified product comprises a crystalline compound of formula (I) according to the method described herein. In a preferred embodiment, the compound of formula (I) is provided as a crystalline solid as described herein.

[0224] Methods of treating diseases, ailments and symptoms This document provides methods for modulating NAD levels in subjects in need, including administration of the compounds, crystalline solids, and / or compositions described herein. Any compound, crystalline solid, or composition described herein may be used to manufacture a medicament for treating any of the diseases or conditions disclosed herein.

[0225] This article provides methods for treating diseases or disorders associated with NAD biosynthesis, including the administration of compounds, crystalline solids and / or compositions described herein.

[0226] This document provides methods for using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used for a variety of therapeutic applications, including, for example, treating and / or alleviating a variety of diseases and disorders, including, for example, diseases or disorders related to aging or stress, diabetes, obesity, neurodegenerative diseases, ataxia and related muscle disorders, acute organ failure, viral symptoms such as cytokine storms, cardiovascular diseases, coagulation disorders, inflammation, cancer, and / or flushing. The method includes administering the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof to a subject in need. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to increase or maintain NAD levels in certain tissues or cells while decreasing NAD levels in other tissues or cells. In various embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to selectively decrease NAD levels in some tissues or cells while decreasing NAD levels in other tissues or cells to a lesser extent.

[0227] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can also be used to treat diseases or conditions associated with inflammation. Exemplary inflammatory conditions include, for example, multiple sclerosis, rheumatoid arthritis, psoriatic arthritis, degenerative joint disease, spondyloarthritis, gouty arthritis, systemic lupus erythematosus, juvenile arthritis, rheumatoid arthritis, osteoarthritis, osteoporosis, and diabetes. For exampleInsulin-dependent diabetes mellitus or juvenile-onset diabetes mellitus, menstrual pain, cystic fibrosis, inflammatory bowel disease, irritable bowel syndrome, Crohn's disease, mucinous colitis, ulcerative colitis, gastritis, esophagitis, pancreatitis, peritonitis, Alzheimer's disease, shock, ankylosing spondylitis, gastritis, conjunctivitis, pancreatitis (acute or chronic), multiple organ injury syndrome (MIS) For example Secondary to sepsis or trauma), myocardial infarction, atherosclerosis, stroke, reperfusion injury ( For example Due to cardiopulmonary bypass or kidney dialysis), acute glomerulonephritis, vasculitis, heat injury (i.e., sunburn), necrotizing enterocolitis, granulocyte transfusion-related syndrome, and / or Sjögren's syndrome. Exemplary inflammatory skin conditions include, for example, eczema, atopic dermatitis, contact dermatitis, urticaria, scleroderma, psoriasis, and skin diseases with acute inflammatory components.

[0228] In other embodiments, the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof may be used to treat skin conditions. Exemplary skin conditions treatable according to the methods described herein include disorders or diseases associated with or caused by inflammation, sunburn, or natural aging. For example, the compositions have been found to treat contact dermatitis (including irritant contact dermatitis and allergic contact dermatitis), atopic dermatitis (also known as allergic eczema), actinic keratosis, keratinization disorders (including eczema), bullous epidermolysis, exfoliative dermatitis, seborrheic dermatitis, erythema (including erythema multiforme and erythema nodosum), damage caused by sunlight or other light sources, discoid lupus erythematosus, dermatomyositis, psoriasis, skin cancer, and the effects of natural aging. In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat wounds and / or burns to promote healing, including, for example, first-, second-, or third-degree burns and / or thermal, chemical, or electrical burns.

[0229] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can also be administered to subjects with acute illnesses (such as organ or tissue injury), such as subjects with stroke or myocardial infarction, or subjects with spinal cord injury, or subjects who have received solid organ transplants (such as liver or kidney). In some embodiments, the compounds, their crystalline solids, and pharmaceutical compositions can be administered to subjects with acute kidney injury (AKI), also known as acute kidney failure (ARF). Kidney function screening can be performed on subjects with AKI or at risk of AKI, for example by testing for abnormal levels of serum creatinine. Subjects can be given prophylactic treatment or treatment in response to acute kidney injury (such as stage 1 AKI). As a form of organ preservation, subjects receiving solid organ transplants can receive prophylactic or post-transplant treatment, or individual organs can be treated in vitro prior to transplantation. Subjects undergoing surgeries other than organ transplantation (such as biopsy or resection or wound repair) can receive prophylactic or post-operative treatment.

[0230] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may also be used in subjects who have or may have chronic damage or chronic disease of solid organs such as the kidneys or liver. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to subjects with chronic kidney disease, such as end-stage renal failure, or nephropathy, or diabetic nephropathy. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to subjects with chronic liver disease, such as chronic infection, cirrhosis, or liver cancer, to repair or limit further damage to the liver. In some embodiments, the crystalline solids and pharmaceutical compositions thereof may be administered to repair alcoholic liver disease, or to stabilize or repair damage resulting from nonalcoholic steatohepatitis (NASH) or nonalcoholic fatty liver disease (NAFLD).

[0231] In some embodiments, the compounds, crystalline solids, or pharmaceutical compositions disclosed herein may be used to treat or prevent diseases or conditions induced or exacerbated by cellular senescence in a subject; methods for reducing the rate of aging in a subject, e.g., after the onset of aging; methods for prolonging the lifespan of a subject; methods for treating or preventing lifespan-related diseases or conditions; methods for treating or preventing diseases or conditions related to cell proliferation capacity; and methods for treating or preventing diseases or conditions caused by cell damage or death. In some embodiments, the method does not work by reducing the incidence of diseases that shorten the lifespan of a subject. In some embodiments, the method does not work by reducing the mortality rate caused by diseases such as cancer.

[0232] In some embodiments, the compounds, crystalline solids, or pharmaceutical compositions disclosed herein may be administered to subjects to generally prolong the lifespan of their cells and protect them against stress and / or apoptosis. Treating subjects with the compounds or crystalline solids described herein can be analogous to subjecting subjects to the stimulant effect of a toxic substance, i.e., a mild stress that is beneficial to the organism and can prolong its lifespan.

[0233] In other embodiments, this document provides a method for treating cardiovascular diseases by administering the disclosed compounds, crystalline solids, and / or pharmaceutical compositions thereof to a subject in need. Cardiovascular diseases that can be treated using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include cardiomyopathy or myocarditis; such as idiopathic cardiomyopathy, metabolic cardiomyopathy, alcoholic cardiomyopathy, drug-induced cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy. The compositions and methods described herein can also be used to treat atherosclerotic disorders (large vessel diseases) of major blood vessels, such as the aorta, coronary arteries, carotid arteries, cerebral arteries, renal arteries, iliac arteries, femoral arteries, and popliteal arteries. Other vascular diseases that can be treated include those related to platelet aggregation, retinal arterioles, glomerular arterioles, nerve-feeding vessels, cardiac arterioles, and associated capillary beds of the eyes, kidneys, heart, and central and peripheral nervous systems. The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can also be used to increase HDL levels in an individual's plasma.

[0234] The disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be administered to subjects who have recently received or may receive a dose of radiation or toxins. In one embodiment, the dose of radiation or toxin is received as part of a work-related or medical procedure, such as working in a nuclear power plant, piloting an aircraft, X-ray, CAT scan, or administration of radioactive dyes for medical imaging; in this embodiment, the compound or crystalline solid is administered as a precautionary measure. In other embodiments, the radiation or toxin exposure is unintentional, for example, due to an industrial accident, residence in a location with natural radiation, acts of terrorism, or acts of war involving radioactive or toxic materials. In such cases, it is preferable to administer the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof as soon as possible after exposure to inhibit apoptosis and the subsequent development of acute radiation syndrome.

[0235] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof can be used to treat age-related conditions such as cancer. Exemplary cancers that can be treated with the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include those of the brain and kidneys; hormone-dependent cancers, including breast cancer, prostate cancer, testicular cancer, and ovarian cancer; lymphoma and leukemia. Other conditions that can be treated include autoimmune diseases such as systemic lupus erythematosus, scleroderma, and arthritis, in which autoimmune cells should be eliminated.

[0236] Viral infections, such as herpes, HIV, adenovirus, and HTLV-1-associated malignant and benign diseases, can also be treated by applying the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof.

[0237] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat patients with infectious diseases such as COVID-19 and other viral infections, including those experiencing symptoms such as cytokine release syndrome (cytokine storm). In some embodiments, the compounds, crystalline solids, and pharmaceutical compositions thereof alleviate or prevent cytokine storms without necessarily treating the underlying viral infection (e.g., COVID-19). Cytokine release syndrome is an acute systemic inflammatory syndrome that can be caused by a variety of factors. In particular, cytokine storms have been described in COVID-19 and other severe viral syndromes (SARS, MERS). A subset of patients exhibit significantly elevated cytokines, and severely ill patients also exhibit much higher levels of IL6, CRP, ferritin, D-dimer, and other markers, as well as lymphopenia (decreased CD4+ and CD8+ T cell counts). For example, in one report, a D-dimer level > 2.0 ug / ml at admission identified a subgroup of patients likely to die (12 / 67 >= 2.0 vs. 1 / 267 < 2.0, sensitivity 92.3%, specificity 83.3%) (“D-dimer levels on admission to predict in-hospital mortality in patients with Covid-19” Zhang L, Yan X, Fan Q et al.) Journal of Thrombosis and Hemostasis (J Thromb Haemost) (April 19, 2020). NAD regulates the release of IL-1β from the NLRP3 inflammasome, thereby regulating cytokine storms. It is known that NAD levels decline with age, which may also contribute to a worse prognosis in older COVID-19 patients. One aspect of this disclosure provides a method for treating COVID-19 in a human patient, comprising administering to the patient the disclosed compound, crystalline solid, or pharmaceutical composition thereof without the administration of zinc sulfate, betaine, or mixtures thereof.

[0238] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat patients with neurodegenerative diseases and traumatic or mechanical injuries to the central nervous system (CNS) or peripheral nervous system (PNS). Examples of neurodegenerative diseases include, but are not limited to, ataxia, Alzheimer's disease (AD), dementia other than Alzheimer's disease, Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), diffuse Lewy body disease, chorea-acanthocytosis, primary lateral sclerosis, multiple sclerosis (MS), eye diseases (ocular neuritis), spinal muscular atrophy, chemotherapy-induced neuropathy (e.g., from vincristine, paclitaxel, bortezomib), diabetes-induced neuropathy, and Friedrich's ataxia.

[0239] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat skeletal muscle disorders, muscle disorders, and conditions including muscle loss, atrophy, and sarcopenia.

[0240] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to reduce appetite and / or increase satiety, thereby resulting in weight loss or preventing weight gain. Subjects requiring this treatment may be overweight, obese, or likely to become overweight or obese.

[0241] In other embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat subjects with cachexia or subjects who may develop cachexia. One method may further include monitoring the subject's disease status. Methods for promoting appetite and / or weight gain may include, for example, pre-identifying subjects who require reduced fat or lipid metabolism, e.g., by weighing the subject and determining the subject's BMI. The method may also include monitoring the subject, for example, during and / or after administration of the disclosed compounds, crystalline solids, or pharmaceutical compositions thereof. Administration may include one or more doses, e.g., in pellet form or continuous delivery. Monitoring may include assessing hormones or metabolites. Exemplary hormones include leptin, adiponectin, resistin, and insulin. Exemplary metabolites include triglycerides, cholesterol, and fatty acids.

[0242] In some embodiments, the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may be used to treat metabolic disorders such as insulin resistance, prediabetes, type 2 diabetes, and / or its complications. Administration of the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof may increase insulin sensitivity and / or decrease insulin levels in a subject. Subjects requiring this treatment may be subjects with insulin resistance or other prodromal symptoms of type 2 diabetes, subjects with type 2 diabetes, or subjects who may develop any of these conditions. For example, a subject may be a subject with insulin resistance, such as a subject with high circulating insulin levels and / or associated conditions such as hyperlipidemia, lipogenic disorders, hypercholesterolemia, impaired glucose tolerance, hyperglycemia, other manifestations of syndrome X, hypertension, atherosclerosis, and lipodystrophy.

[0243] This article provides a method for regulating blood glucose concentration in mammals. As used herein, regulating blood glucose concentration refers to any increase, decrease, and / or maintenance of blood glucose concentration compared to a previously determined level.

[0244] The therapeutic methods disclosed in this article also involve methods for modulating the circadian rhythm clock, thereby modulating or influencing biological functions regulated (sometimes referred to as its influence, subordinate to, or mediated by) the activity of the circadian rhythm clock. Typically, these biological functions exhibit patterns of activity and inactivity that usually repeat approximately every 24 hours, oscillating between “active” and “inactive” states during the 24-hour period.

[0245] Therefore, the present invention provides a method for modulating the activity of the circadian rhythm clock by administering the disclosed compounds, crystalline solids, or pharmaceutical compositions to mammals in need. Typically, the regulation of circadian rhythm clock activity is a result of circadian clock:BMAL1 regulation, which is achieved according to the method of the present invention by modulating the activity of SIRT1. SIRT1 activity is typically modulated according to the method of the present invention by administering the disclosed compounds, crystalline solids, or pharmaceutical compositions, and in some embodiments by administering compounds or crystalline solids that affect the NAD pathway. Regulation of the circadian rhythm clock thus allows for the modulation of activities mediated by the circadian rhythm clock.

[0246] According to the present invention, the activity of the circadian rhythm clock can be increased, decreased, or maintained by applying the compounds, crystalline solids, or pharmaceutical compositions disclosed herein. Therefore, biological functions (sometimes referred to as biological activities) regulated by the activity of the circadian rhythm clock can also be increased, decreased, or maintained. Furthermore, these biological functions can also be time-shifted; that is, activities that typically occur during specific time periods, such as, for example, during daytime or diurnal (sometimes referred to as the light cycle) or during nighttime or twilight (sometimes referred to as the dark cycle), can be shifted so that the activity instead occurs during the dark cycle or light cycle, respectively.

[0247] In various embodiments, this document discloses methods for differentially modulating nicotinamide adenine dinucleotide (NAD) levels in two or more tissue or cell types. Such methods may include administering a compound, crystalline solid, or composition disclosed herein, wherein said administration induces a differential response in NAD levels in a first tissue or cell type compared to a second tissue or cell type. In various implementation schemes, the differential response to NAD levels is selected from at least 10% NAD level difference, at least 20% NAD level difference, at least 30% NAD level difference, at least 40% NAD level difference, at least 50% NAD level difference, at least 60% NAD level difference, at least 70% NAD level difference, at least 80% NAD level difference, at least 90% NAD level difference, at least 100% NAD level difference, at least 200% NAD level difference, at least 300% NAD level difference, at least 400% NAD level difference, at least 500% NAD level difference, at least 600% NAD level difference, at least 700% NAD level difference, at least 800% NAD level difference, at least 900% NAD level difference, and at least 1000% NAD level difference. In various embodiments, the differential response of NAD levels compared to untreated NAD levels or pre-treatment NAD levels is an increase of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% in NAD levels in a first tissue or cell type, and a simultaneous decrease of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% in NAD levels in a second tissue or cell type compared to untreated NAD levels or pre-treatment NAD levels. In various embodiments, the differential response to NAD levels is that, compared to untreated NAD levels, the NAD level in the first tissue or cell type is maintained within 10%, and the NAD level in the second tissue or cell type is simultaneously reduced by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% compared to untreated NAD levels.In various embodiments, the differential response to NAD levels is a reduction of at least 10% in NAD levels in a first tissue or cell type compared to untreated NAD levels, and a simultaneous reduction in NAD levels in a second tissue or cell type compared to untreated NAD levels, wherein the reduction in the second tissue or cell type is at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or 1000% greater than the reduction in the first tissue or cell type. In various embodiments, the first tissue or cell type is normal tissue or cells, and the second tissue or cell type is neoplastic or cancerous.

[0248] The cancer treatment methods disclosed herein include treatment for individuals in need. Exemplary cancers that can be treated using the disclosed compounds, crystalline solids, and pharmaceutical compositions thereof include those of the brain and kidneys; hormone-dependent cancers, including breast cancer, prostate cancer, testicular cancer, and ovarian cancer; lymphoma and leukemia. In various embodiments, the cancer may be a type of cancer common in men, such as lung cancer, prostate cancer, colorectal cancer, and stomach cancer. In various embodiments, the cancer may be a type of cancer common in women, such as breast cancer, colorectal cancer, lung cancer, and cervical cancer. In various embodiments, the cancer may be skin cancer, such as melanoma, squamous cell carcinoma, or basal cell carcinoma. In various embodiments, the cancer may be a type of cancer common in children, such as acute lymphoblastic leukemia, brain tumors, or non-Hodgkin's lymphoma. In various embodiments, the method exhibits selective cell inhibition or cytotoxicity, wherein this effect is demonstrated by a reduced viability of neoplastic or cancerous tissue or cells compared to untreated neoplastic or cancerous tissue or cells.

[0249] The method includes cases where the first tissue or cell type is normal tissue or cells, and the method is a treatment for promoting health or increasing the biological activity of the first tissue or cell type in an individual in need. In various embodiments, the treatment does not result in an increased risk of cancer diagnosis in the treated individual. Preferably, the treatment reduces the risk of cancer diagnosis in the treated individual.

[0250] Various methods include treating or inhibiting cancer in individuals in need, wherein such methods include administering the compounds, crystalline solids, or compositions described herein. In various embodiments, methods are disclosed herein for increasing or maintaining healthy tissue or cells in individuals in need without increasing the risk of neoplastic or cancerous tissue or cell growth, such methods include administering the compounds, crystalline solids, or compositions described herein.

[0251] In various embodiments, this document describes methods for increasing or maintaining healthy tissue or cells in an individual while simultaneously inhibiting the growth of neoplastic or cancerous tissue or cells, such methods comprising administering the compounds, crystalline solids, or compositions described herein. In various embodiments, the disclosed methods include methods for increasing or maintaining nicotinamide adenine dinucleotide (NAD) levels in at least one healthy tissue or cell type, such methods comprising administering the compounds, crystalline solids, or compositions described herein to the healthy tissue or cell type. In various embodiments, this document describes methods for reducing the viability of at least one cancerous tissue or cell type, such methods comprising administering the compounds, crystalline solids, or compositions described herein to the cancerous tissue or cell type.

[0252] Furthermore, the methods described herein include methods for regulating NAD levels in at least one tissue or cell type in a mixture of tissue or cell types, such methods comprising targeted delivery of the compounds, crystalline solids, or compositions described herein to a desired tissue or cell type. In various embodiments, the targeted delivery is non-systemic.

[0253] Compositions and pharmaceutical compositions This document also provides compositions of the disclosed compounds and crystalline solids. In some embodiments, the composition comprises 1) a crystalline solid comprising a compound of formula (I) or formula (II) or a salt thereof, and 2) one or more pharmaceutically acceptable excipients. In other embodiments, the composition comprises 1) a compound of formula (I) or formula (II) or a salt thereof, and 2) one or more pharmaceutically acceptable excipients.

[0254] In some embodiments, the composition is a solution. For example, in some embodiments, a crystalline solid comprising a compound of formula (I) or (II) is dissolved in a solvent or carrier to form a solution of a compound of formula (I) or (II). In a preferred embodiment, the purity of the crystalline solid is such that the resulting solution is pure or substantially pure and / or free from or substantially free from one or more impurities.

[0255] In some preferred embodiments, this disclosure provides compositions comprising a compound of formula (I) or formula (II) or a crystalline solid comprising a compound of formula (I) or formula (II), wherein the composition is pure or substantially pure. In some preferred embodiments, the composition is greater than about 90% pure. More preferably, the composition is greater than about 95% pure, or even more preferably greater than about 98% pure, for example, greater than about 98% pure. In some embodiments, the percentages are by weight.

[0256] In preferred embodiments, the composition contains less than about 5% impurities, less than about 2% impurities, less than about 1% impurities, or less than about 0.5% impurities. For example, in a preferred embodiment of a composition comprising a compound of formula (I), the composition comprises less than about 5% propyl nicotinic acid, less than about 2% propyl nicotinic acid, less than about 1% propyl nicotinic acid, less than about 0.5% propyl nicotinic acid, less than about 0.1% propyl nicotinic acid, or less than about 0.01% propyl nicotinic acid. In a preferred embodiment of a composition comprising a compound of formula (II), the composition comprises less than about 5% nicotinic acid riboside, less than about 2% nicotinic acid riboside, less than about 1% nicotinic acid riboside, less than about 0.5% nicotinic acid riboside, less than about 0.1% nicotinic acid riboside, or less than about 0.01% nicotinic acid riboside. In some embodiments, the percentages are by weight.

[0257] In some embodiments, pharmaceutically acceptable excipients are selected from anti-adhesives, adhesives, coatings, dyes, disintegrants, flavorings, flow aids, lubricants, preservatives, adsorbents, sweeteners, syrups, elixirs, dispersants, diluents, fillers, granulators, coating agents, waxes, suspending agents, wetting agents, thickeners, and mediators, and combinations thereof. In some embodiments, the excipients are solid excipients.

[0258] In some embodiments, the pharmaceutically acceptable excipient is present in an amount of at least about 5% by weight, at least about 10% by weight, at least about 15% by weight, at least about 20% by weight, at least about 25% by weight, at least about 30% by weight, at least about 35% by weight, at least about 40% by weight, at least about 45% by weight, at least about 50% by weight, at least about 55% by weight, or at least about 60% by weight of the composition. In some embodiments, the pharmaceutically acceptable excipient is present in an amount of at least about 20% by weight, at least about 25% by weight, at least about 30% by weight, at least about 35% by weight, or at least about 40% by weight of the composition, preferably at least about 30% by weight. In other embodiments, the pharmaceutically acceptable excipient is present in an amount of at least about 50% by weight of the composition. The pH of the formulation may be in the range of about 3 to about 11, but is typically about 7 to about 10.

[0259] In some embodiments, the composition is in a solid form selected from tablets, pills, capsules, pouches, lozenges, granules, powders, sachets, dry powder inhalation forms, chewable tablets, soft lozenges, and sugar lozenges. In some embodiments, the composition is in the form of tablets. In other embodiments, the composition is in the form of hard or soft capsules.

[0260] The compounds and crystalline solids disclosed herein are formulated with conventional carriers and excipients, which can be selected according to conventional practice. Tablets may contain excipients, gliding agents, fillers, binders, etc. All formulations will optionally contain excipients, such as in… Handbook of Pharmaceutical ExcipientsThose described in (1986). Suitable excipients are also listed in the FDA's Inactive Ingredient Database. Excipients include ascorbic acid and other antioxidants, chelating agents such as EDTA, carbohydrates such as dextran, hydroxyalkyl cellulose, hydroxyalkyl methyl cellulose, stearic acid, etc.

[0261] While the active pharmaceutical ingredient can be administered alone, it is preferred to be in the form of a pharmaceutical formulation. The veterinary and human formulations of the present invention comprise at least one active ingredient as defined above, as well as one or more acceptable carriers and optional other therapeutic ingredients. Some examples of materials that can be used as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose, and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, and corn oil. (10) Soybean oil; (11) Glycols, such as propylene glycol; (12) Polyols, such as glycerol, sorbitol, mannitol and polyethylene glycol; (13) Esters, such as ethyl oleate and ethyl laurate; (14) Agar; (15) Buffers, such as magnesium hydroxide and aluminum hydroxide; (16) Alginate; (17) Pyrogen-free water; (18) Isotonic saline; (19) Ringer's solution; (10) Ethanol; (21) Phosphate buffer solution; and (22) Other non-toxic and compatible substances used in pharmaceutical formulations.

[0262] The pharmaceutical composition (formulation) may be administered to a subject via any of a variety of routes of administration, including, for example, oral (e.g., infusions in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including dispersed capsules and gelatin capsules), granules, powders, pellets, or pastes applied to the tongue); absorption via the oral mucosa (e.g., sublingual); anal, rectal, or vaginal (e.g., as vaginal suppositories, creams, or foams); parenteral (including intramuscular, intravenous, subcutaneous, or intrathecal, e.g., as sterile solutions or suspensions); nasal; intraperitoneal; subcutaneous; percutaneous (e.g., as patches applied to the skin); and topical (e.g., as creams, ointments, or sprays applied to the skin, or as eye drops). The compound or crystalline solid may also be formulated for inhalation. In some embodiments, the crystalline solid of the compound may simply be dissolved or suspended in sterile water. Details of suitable routes of application and compositions applicable thereto can be found, for example, in U.S. Patent Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896 and the patents referenced therein.

[0263] Formulations of this disclosure suitable for oral administration may be in the form of discrete units, such as capsules, sachets, or tablets, each containing a predetermined amount of the active ingredient in powder or granule form. The active ingredient may also be administered as a pellet, syrup, or paste.

[0264] Tablets are made by compression or molding, optionally together with one or more excipients. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (such as powder or granules) optionally mixed with a binder, lubricant, inert diluent, preservative, surfactant, or dispersant in a suitable machine. Molded tablets can be manufactured by molding a mixture of powdered active ingredients moistened with an inert liquid diluent in a suitable machine. Tablets may optionally be coated or scored and optionally formulated to provide a slow or controlled release of the active ingredient from it.

[0265] Pharmaceutical formulations according to this disclosure comprise compounds or crystalline solids according to this disclosure, as well as one or more pharmaceutically acceptable carriers or excipients and optional other therapeutic agents. Pharmaceutical formulations containing an active ingredient can be in any form suitable for the intended method of administration. When intended for oral administration, for example, tablets, lozenges, sugar lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups, or elixirs can be prepared. Compositions intended for oral administration can be prepared according to any method known in the art for manufacturing pharmaceutical compositions, and such compositions may contain one or more pharmaceutical agents, including sweeteners, flavoring agents, coloring agents, and preservatives, to provide a palatable formulation. Tablets containing an active ingredient mixed with a non-toxic, pharmaceutically acceptable excipient suitable for manufacturing tablets are acceptable. These excipients can be, for example, inert diluents such as calcium carbonate or sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrants such as corn starch or alginate; binders such as starch, gelatin, or gum arabic; and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated, or they may be coated using known techniques, including microencapsulation, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a longer duration of action. For example, they may be used alone with a delaying material, such as glyceryl monostearate or glyceryl distearate, or together with a wax.

[0266] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent (such as calcium phosphate or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily medium (such as peanut oil, liquid paraffin, or olive oil).

[0267] The aqueous suspensions disclosed herein may contain active substances mixed with excipients suitable for preparing aqueous suspensions. Such excipients include suspending agents such as sodium carboxymethyl cellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and gum arabic; and dispersing or wetting agents such as natural phospholipids (e.g., lecithin), condensation products of alkyl esters and fatty acids (e.g., polyoxyethylene stearate), condensation products of ethylene oxide and long-chain fatty alcohols (e.g., heptadecanethoxyethanol), and condensation products of ethylene oxide and esters derived from fatty acids and hexadiol anhydrides (e.g., polyoxyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives (such as ethylparaben or n-propylparaben), one or more colorants, one or more flavoring agents, and one or more sweeteners (such as sucrose or saccharin). The liquid formulation may also include eye drops or other forms delivered to the surface of the eye or adjacent sites such as the lacrimal glands. Liquid formulations may include intravenous formulations, excipients and carriers, such as saline solutions or buffer solutions, and packaging or containers for such formulations for injection or infusion.

[0268] By adding water, the dispersible powders and granules suitable for preparing the aqueous suspensions of this disclosure provide an active ingredient that can be mixed with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are exemplified by those disclosed above. Other excipients, such as sweeteners, flavoring agents, and coloring agents, may also be present.

[0269] The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will vary depending on the subject being treated and the specific administration method. For example, a sustained-release formulation for oral administration in humans may contain about 1 to about 1000 mg of active material combined with a suitable and appropriate amount of carrier material, the amount of which may vary from about 5% to about 95% (weight:weight) of the total composition. Pharmaceutical compositions can be prepared to provide easily measurable dosage amounts.

[0270] Although the dosage will vary depending on the patient's symptoms, age and weight, the nature and severity of the condition to be treated or prevented, the route of administration, and the form of the drug, generally, for adult human patients, a daily dose of the compound or crystalline solid is recommended to be from 0.01 to 3000 mg, and this can be administered as a single dose or in divided doses. Generally, the compositions disclosed herein can be provided in an aqueous solution containing about 0.1-30% w / v of the compound or crystalline solid disclosed herein, as well as other substances, for parenteral administration. Typical dosage ranges are from about 0.01 to about 50 mg / kg body weight daily, administered as a single dose or in 2-4 divided doses. In some embodiments, the compound and / or crystalline solid described herein are administered in amounts from about 1 to about 3000 mg daily, from about 100 to about 1000 mg daily, or from about 250 to about 750 mg daily. If desired, the effective daily dose of the active compound or crystalline solid can be divided into one, two, three, four, five, six, or more sub-dose administrations at appropriate time intervals throughout the day, optionally administered in a unit dosage form. In some embodiments, the compounds and / or crystalline solids described herein are administered once, twice, three times, four times, five times, six times, or more daily. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will typically be the amount of the compound and / or crystalline solid that produces the therapeutic effect.

[0271] Formulations suitable for intrapulmonary or intranasal application have particle sizes, such as about 0.5, about 1, about 30, or about 35 micrometers, in the range of about 0.1 to about 500 micrometers, and are administered via rapid inhalation through the nasal passages or oral inhalation to reach the alveolar sacs. Suitable formulations include aqueous or oily solutions of the active ingredient. Formulations suitable for aerosol or dry powder application can be prepared according to conventional methods and can be delivered together with other therapeutic agents.

[0272] The formulation exists in single-dose or multi-dose containers, such as sealed ampoules and vials, and can be stored under lyophilized (freeze-dried) conditions where a sterile liquid carrier (e.g., water for injection) needs to be added immediately before use. Temporary injectable solutions and suspensions are prepared from the aforementioned types of sterile powders, granules, and tablets. Preferred single-dose formulations are those containing a daily dose or a sub-daily dose, or an appropriate fraction thereof of the active ingredient as described above.

[0273] It should be understood that, in addition to the ingredients specifically mentioned above, formulations of this disclosure may include other pharmaceutical agents conventional in the art related to the type of formulation discussed, such as flavoring agents suitable for oral administration.

[0274] In some embodiments, the amount of the compound and / or crystalline solid in the composition is from about 0.001% by weight to up to 100% by weight.

[0275] In some embodiments, the compound and / or crystalline solid is the sole active pharmaceutical ingredient in the composition. Alternatively, the compound and / or crystalline solid is formulated into a composition with one or more additional active pharmaceutical ingredients. When formulated as the sole active pharmaceutical ingredient, the compound and / or crystalline solid may be administered alone or as part of a regimen having one or more separately formulated active pharmaceutical ingredients.

[0276] When administered in combination in the same formulation or as part of a regimen having one or more separately formulated active pharmaceutical ingredients, the additional active pharmaceutical ingredient may be selected from compounds in the NAD pathway, such as nicotinic acid (NA), nicotinamide (Nam), nicotinamide mononucleotide (NMN), nicotinamide riboside (NR), nicotinic acid riboside (NAR), nicotinamide adenine dinucleotide (NAD / NADH), nicotinamide adenine dinucleotide phosphate (NADP), and nicotinamide adenine dinucleotide (NaAD). In some embodiments, compounds of formulas I and II are administered in combination. In some embodiments, the additional active pharmaceutical ingredient is an amorphous solid. In some embodiments, the additional active pharmaceutical ingredient is a crystalline solid. In some embodiments, the additional active pharmaceutical ingredient is amorphous NMN. In some embodiments, the additional active pharmaceutical ingredient is crystalline NMN.

[0277] The invention described herein in general will be more readily understood by referring to the following examples, which are included only for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[0278] Example Example 1.1: Preparation of Compound 2 1.00 g (3.92 mmol) of nicotinic acid riboside was packed into a 50 mL recovery flask and purged with argon. 6 mL of trimethyl phosphate was added, and the mixture was stirred and cooled in an ice bath. Next, 0.73 mL (7.84 mmol) of phosphoryl chloride was added. After 30 minutes, the reaction was confirmed to be complete by LC / MS. The reaction mixture was added dropwise to 10 mL of ice-cold water. After the addition was complete, the mixture was concentrated under vacuum to remove approximately 9 g of solvent. The concentrated mixture was then added dropwise to a stirred, ice-cold solution of 3.2 mL (23.0 mmol) triethylamine in 50 mL of 1-propanol, yielding a mixture with pH = 3 (determined by pH paper). The suspension was stirred for 1 h, and the solid precipitate was filtered and washed with 1-propanol to give the first batch of crude compound 2. After filtration, a solid formed in the supernatant. The filtrate was stirred for 72 h, and the suspension was filtered, and the precipitate was washed with 1-propanol to give the second batch of crude compound 2. The second batch was vacuum dried, weighing 1.00 g, but was still moist. 1 H and 31 P NMR analysis showed that the second batch was of better purity than the first. The second batch was used as seed material for subsequent experiments. See also Figure 4 .

[0279] 1 H-NMR (500 MHz; D2O): δ9.42 (s, 1H), 9.25 (d, J = 6.3 Hz, 1H), 9.00(d, J = 8.0 Hz, 1H), 8.22 (dd, J = 7.8, 6.5 Hz, 1H), 6.16 (d, J = 5.3 Hz,1H), 4.58-4.56 (m, 1H), 4.49 (t, J = 5.1 Hz, 1H), 4.37 (dd, J = 5.0, 2.8 Hz,1H), 4.26-4.22 (m, 1H), 4.12-4.08 (d,d,d 1H, J = 12.0, 5.1, 2.2 Hz) Example 1.2A: Crystallization of Compound 2 Add 300 μL of water to 100 mg of amorphous compound 2 (prepared separately). Add 350 μL of 1-propanol to obtain a turbid solution. Use a second batch of compound 2 from the above experiment to seed the mixture. After several hours, crystals formed in a vial. The mixture was not filtered and remained as a slurry.

[0280] Example 1.2B: Crystallization of Compound 2 2.00 g of amorphous compound 2 (prepared separately) was dissolved in 6 mL of water, and then 7 mL of 1-propanol was added to obtain a turbid solution. Another 2 mL of water was added to obtain a clear solution. The mixture was seeded using a second batch of compound 2 from the previous experiment, but no crystals formed. 0.2 mL of n-propanol was added to obtain a turbid mixture. This was stirred for one day, and no crystals were obtained. Then, the mixture was seeded with a drop of slurry from the second experiment to prepare compound 2 crystals, resulting in rapid crystallization. This was stirred at ambient temperature for two days. The solid was filtered and washed with 15 mL of (2:1 v:v) 1-propanol:water, 15 mL of 1-propanol, and then 2 x 15 mL of methyl tert-butyl ether. The sample was dried under high vacuum at ambient temperature for 1 hour to obtain 1.92 g (96% mass recovery) of white solid. The water solubility of the isolated product was approximately 20 mg / mL (50:1 w:w water:compound 2), compared to the fact that the starting compound 2 was freely soluble in water (3:1 water:compound 2).

[0281] Example 1.2C: Crystallization of Compound 2 500 mg of amorphous compound 2 was dissolved in 1.5 mL of water. The sample dissolved completely, and crystals began to form. The crystals were allowed to grow without stirring. The water was gently poured off, and the solid was dried under high vacuum. Single-crystal X-ray diffraction-quality crystals of compound 2 were prepared. These crystals were used to generate the XRPD signature of compound 2 crystals (Figure 2) and also to obtain the single-crystal X-ray structure of compound 2. Figure 2C ).

[0282] Example 2.1: Preparation of Compound 1 Compound 2 was filtered from 1-propanol before use and packed into a 500 mL recovery bottle, which was then purged with argon. The solid was suspended in 1-propanol and cooled in an ice bath. HCl gas was bubbled into the reaction mixture. The solid dissolved upon bubbling into the suspension. The reaction mixture was removed from the ice bath and stirred at room temperature for more than three days. When no starting material residue was confirmed by LC / MS, the reaction mixture was concentrated on a rotary evaporator until an oil was obtained. An aliquot was placed under high vacuum for 30 minutes. No foaming occurred in the aliquot. This sample was used for LC / MS. The oil was diluted with 10 mL of 1-propanol and cooled in an ice bath. A total of 1.5 mL of triethylamine was added to bring the pH to 4 to 5. The addition of triethylamine resulted in the formation of a large amount of precipitate. 40 mL of 1-propanol was added to dissolve all the precipitate. Seed crystals of Compound 1 were added to the solution. Over time, the solution became cloudy and precipitate formation was observed. The suspension was stirred overnight at room temperature. After 4 hours under high vacuum, aliquots of the sample were used for LC / MS. The contents of the flask appeared more crystalline and filterable compared to the “emulsion” suspension. The solid was filtered. The filter cake was washed twice with 10 mL of 1-propanol, followed by two washes with 10 mL of MTBE each time. A sample of the 1-propanol filtrate was used for LC / MS. The solid was transferred to a vial. The wet weight was approximately 1.2 g. The sample was placed under high vacuum for 1 hour. 1.23 g (55% yield) of white solid was obtained. The sample was used for LC / MS. 1 H and 31 P NMR. See also Figure 3 .

[0283] Example 2.2A: Polymorph Screening The polymorphism of amorphous compound 1 was screened according to the conditions provided in Table 1. The treatment was carried out between observation 1 and observation 2, including a 4-day aging cycle between 0℃ (1 hr) and -20℃ (7 hrs). The results are shown in the figure.

[0284] Table 1: x = suspension Example 2.2B: Salt Screening Polymorphs of amorphous compound 1 were screened according to Table 2. Compound 1 (15 mg) was weighed into an HPLC vial and a magnetic stir bar was added. The compound was dissolved in approximately 15 vol (200 µl) of EtOH at 5 °C with stirring at 500 rpm. After dissolution, a one-molar equivalent of the counterion was added, and the solution was stirred at 5 °C (45 µl of 1 M stock solution or 90 µl of 0.5 M stock solution). The sample was cooled to -20 °C at 1 °C / min, but still in solution. The solution was slowly evaporated at 5 °C through a needle in the vial cap.

[0285] Table 2: Note: P = solution, N / P = not executed Example 2.2C: Eutectic Screening Polymorphs of amorphous compound 1 were screened according to Table 3. Compound 1 (25 mg) was weighed into an HPLC vial and two grinding balls were added. A 1 mol equivalent of the co-formed product (as a solid) was added. The mixture was initially milled at 500 rpm for 2 hours on a Fritsch planetary mill, and the recovered solid was analyzed by XRPD. No crystals were obtained, and the resulting solid was moistened with a drop of THF (7.5 µl) and milled at 500 rpm for 2 hours on a Fritsch planetary mill. The milling was observed, and the recovered solid was subjected to XRPD.

[0286] Table 3: Note: P = solution, N / P = not executed Example 2.2D: Crystal form of compound 1 Compound 1 (150 mg) was dissolved in 10 vol (1.5 ml) of pure EtOH at room temperature with stirring. After 3 minutes, precipitate formation began. The sample was stirred for another 15 minutes, then filtered and dried under positive pressure. The sample was then placed in a vacuum oven under vacuum and kept at room temperature for 30 minutes. Then, prior to characterization, the sample was placed overnight in a fume hood in a vial capped with perforated aluminum foil. Figure 1A Obtain the XPD map. See also: Figure 1B Percentage yield = approximately 63% obtained by HPLC and purity 98.7%. After storage at 40°C / 75% RH for seven days, the material is a viscous solid with a purity of 96.4% obtained by HPLC. The sample is a fine white powder stable at room temperature; a small amount of solvent is retained, but analysis indicates that the sample is an anhydrous, non-solventized solid. The sample is stable at humidity conditions up to 70% RH.

[0287] Figure 1C This is the representation of the lattice cell of compound 1, which has the following properties: Data for compound 1 were obtained via single-crystal X-ray diffraction, and the structure was solved using a direct method, corrected using least-squares correction. The atomic distribution and positions in the crystal structure were specified based on the electron density observed in the Fourier difference plot, which converged to a model that fit the experimental data well. Non-hydrogen atoms were anisotropically corrected, giving anisotropic displacement parameters (thermal ellipsoids), which can be seen in the ORTEP images. Figure 1C ).

[0288] Example 2.2E: Magnification of the crystal form of compound 1 Amorphous compound 1 (1.23 g) was weighed and placed in a 20 ml vial, and treated with 7 vol (8.60 ml) of methanol to obtain a clear solution. The solution was stirred at 400 rpm at 35 °C on a 'Polar Bear' instrument. The solution was supersaturated by adding 0.5 vol (615 µl) of TBME, and then seeded with the previously crystallized material (approximately 60 mg) retained in solution. The sample was then cooled to 25 °C at 0.1 °C / min before adding the antisolvent. 2.6 vol (3.2 ml) of TBME was added over 50 minutes (1 µl / sec) using a syringe pump. The suspension was then cooled to 5 °C at 0.1 °C / min. The sample was held at 5 °C for one hour before separation under vacuum using a Buchner funnel. The sample was dried under vacuum for 20 mins, and then further dried overnight in a vacuum oven. Figure 1A Obtain the XPD map. See also: Figure 1B Percentage yield = approximately 45% obtained by HPLC and purity 97.9%.

[0289] Example 3: Stability study of compound 2 Amorphous and crystalline forms of compound 2 were prepared and stored under stress conditions to compare the stability of their respective forms, as shown in Table 4. At T=0, the AUC of amorphous compound 2 was 97.3%, and the AUC of crystalline compound 2 was 99%. Samples were sealed in wide-mouth flasks containing saturated salt solutions to generate the required relative humidity: ammonium nitrate for RH = 60%; sodium chloride for RH = 75%; and saturated potassium nitrate for RH = 97%. Solid phosphorus pentoxide was used for RH = 0%. Waters Atlantis T3 C was used. 18HPLC data were collected using an Agilent 1290 system with a 3 μm, 100 x 4.6 mm column and an in-line guard column. Mobile phase A: 200 mM ammonium carbonate (pH 3.8); Mobile phase B: 95:5 MeOH:Mobile phase A. The pump rate was 1 mL / min. The gradient was 0% B for 5 min, followed by a 20 min gradient to 100% B, and finally held for 3 min. Data were collected at 254 nm via DAD. Legend: AUC = Area under the curve at 254 nm; RH = Relative humidity; OS = Out of scale The results in Table 4 show that the crystalline form of compound 2 has improved stability compared to the amorphous form.

[0290] By incorporation and equivalent forms All publications and patents mentioned herein are hereby incorporated in their entirety by reference as if each individual publication or patent were specifically and individually indicated by reference. In the event of conflict, all definitions contained herein shall prevail.

[0291] While specific embodiments of the invention have been discussed, the foregoing description is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon reading this specification and the following claims. The full scope of the invention should be determined by reference to the full scope of the claims and their equivalents, the specification, and these variations.

Claims

1. A crystalline solid comprising compound 1: 。 2. The crystalline solid according to claim 1, having 2θ values ​​of 16.1, 20.1, and 24.

5.

3. The crystalline solid according to any one of claims 1 to 2, wherein the solid is anhydrous.

4. The crystalline solid according to any one of claims 1 to 3, wherein the solid is selected from methanol solvates, ethanol solvates, 1-propanol solvates, 2-propanol solvates, C-4 alcohol solvates, C-5 alcohol solvates and C-6 alcohol solvates, preferably the methanol solvates.

5. The crystalline solid according to any one of claims 1 to 4, wherein the crystalline solid contains residual non-solventized solvent or residual non-hydrated water.

6. The crystalline solid according to any one of claims 1 to 5, comprising less than about 5% by weight of propyl nicotinic acid, preferably less than about 1% by weight of propyl nicotinic acid.

7. The crystalline solid according to any one of claims 1 to 4, comprising less than about 5% by weight of compound 2: 。 8. The crystalline solid according to claim 7, comprising less than about 1% by weight of compound 2.

9. The crystalline solid according to any one of claims 1 to 8, comprising at least about 90% by weight of compound 1.

10. The crystalline solid according to claim 9, comprising at least about 95% by weight of compound 1.

Images