HK1238251B - Crystalline form of nicotinamide riboside - Google Patents

Description

Crystalline form of nicotinamide riboside

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No. 62/028,702, filed July 24, 2014, the entire contents of which are incorporated herein by reference.

Technical Field

The present disclosure relates to crystalline forms of nicotinamide riboside, particularly crystalline forms of nicotinamide riboside chloride, as well as compositions containing and methods of using the crystalline forms.

background

Crystalline forms of useful molecules can have advantageous properties relative to amorphous forms of such molecules. For example, crystalline forms are generally easier to handle and process, for example, when preparing compositions containing the crystalline forms. Crystalline forms generally have greater storage stability and are easier to purify. The use of crystalline forms of pharmaceutically useful compounds can also improve the performance characteristics of pharmaceutical products containing the compounds. Obtaining crystalline forms is also used to expand the catalog of materials that formulation scientists can use to optimize formulations, for example, by providing products with different properties, for example, better processing or handling characteristics, improved dissolution profiles, or improved shelf life.

Nicotinamide riboside (CAS number: 1341-23-7) is a precursor to nicotinamide adenine dinucleotide (NAD) and represents a source of vitamin B3. Recent studies have shown that consuming larger amounts of nicotinamide riboside than naturally found in food can produce novel health benefits. For example, nicotinamide riboside has been associated with increasing tissue NAD concentrations, inducing insulin sensitivity, and enhancing sirtuin function. See Chi Y et al., Curr Opin Clin Nutr Metab Care. 2013 Nov;16(6):657-61. Its ability to increase NAD production suggests that nicotinamide riboside may also improve mitochondrial health, stimulate mitochondrial function, and induce the production of new mitochondria. Other studies of nicotinamide riboside in models of Alzheimer's disease have shown that the molecule is bioavailable to the brain and provides neuroprotection, possibly through stimulation of brain NAD synthesis. Additionally, a 2012 study observed that mice fed a high-fat diet supplemented with nicotinamide riboside gained 60% less weight than mice fed the same high-fat diet without nicotinamide riboside.

Nicotinamide riboside chloride (3-carbamoyl-1-[(2R,3R,4S5R)-3,4-dihydroxy-5-(hydroxymethyl)oxolan-2-yl]-pyridin-1-ium chloride; also known as 1-(β-D-ribofuranosyl)nicotinamide chloride), is a known salt form of nicotinamide riboside and has the structure depicted below:

Despite the useful properties of nicotinamide riboside and its chloride salt, for example, as pharmaceuticals or nutritional supplements, and the benefits of providing this molecule in an ordered form, improvements are often desired.

Overview

The present disclosure relates to crystalline forms of nicotinamide riboside, including Form I of nicotinamide riboside chloride according to Formula I

.

Also disclosed are pharmaceutical compositions containing crystalline Form I of nicotinamide riboside chloride, and methods of preparing such pharmaceutical compositions.

In other aspects, the present disclosure relates to a method comprising administering crystalline Form I of nicotinamide riboside chloride to a subject.

The present disclosure also provides a method for preparing crystalline Form I of nicotinamide riboside chloride. Also provided is crystalline Form I of nicotinamide riboside chloride prepared according to any disclosed method for preparing crystalline Form I.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG1 provides an X-ray powder diffraction pattern of crystalline nicotinamide riboside chloride.

Figure 2 shows the solid-state IR spectrum of crystalline nicotinamide riboside chloride.

FIG3 depicts scanning electron microscopy (SEM) images of two different morphologies of crystalline nicotinamide riboside chloride.

Figure 4 provides DSC thermograms of crystalline Form I of nicotinamide riboside chloride measured at various experimental heating rates.

Figure 5 provides a DSC thermogram of a sample of crystalline Form I of nicotinamide riboside chloride at a heating rate of 10 K/min.

FIG6 provides a DSC thermogram of an amorphous form of nicotinamide riboside.

FIG7A shows a TGA/SDTA thermogram of crystalline Form I of nicotinamide riboside chloride, and FIG7B shows a TGA/SDTA thermogram of an amorphous form of nicotinamide riboside.

FIG8A provides a DVS mass change graph of a sample of crystalline Form I of nicotinamide riboside chloride, and FIG8B provides a DVS isotherm graph of a sample of crystalline Form I of nicotinamide riboside chloride.

FIG9A provides a DVS mass change graph of a sample of the amorphous form of nicotinamide riboside chloride, and FIG9B provides a DVS isotherm graph of a sample of the amorphous form of nicotinamide riboside chloride.

Figure 10 provides a comparison of the absorbance curve of crystalline Form I of nicotinamide riboside chloride and the absorbance curve of an amorphous sample of nicotinamide riboside chloride.

Detailed Description of Illustrative Embodiments

The present invention may be more readily understood by reference to the following detailed description taken in conjunction with the accompanying drawings and examples, which form a part of this disclosure. It should be understood that the present invention is not limited to the specific products, methods, conditions or parameters described and/or shown herein, and that the terminology used herein is only for the purpose of describing specific embodiments by way of example and is not intended to limit the invention as claimed.

The entire disclosure of each patent, patent application, and publication cited or described herein is hereby incorporated by reference.

Unless otherwise indicated, the following terms and abbreviations used above and throughout the disclosure shall be understood to have the following meanings.

Unless the context clearly indicates otherwise, in this disclosure, the singular forms "a," "an," and "the" include plural referents, and references to specific values include at least that specific value. Thus, for example, reference to "a solvent" refers to one or more such solvents, as well as equivalents thereof known to those skilled in the art, and so forth. Furthermore, when it is indicated that an element "can be" X, Y, or Z, such usage is not intended to exclude other options for the element in all cases.

When an approximation is used to express a value, the use of "about" as used above can be understood to mean that the specific value forms another embodiment. As used herein, "about X" (where X is a value) preferably refers to ±10% of the recited value, inclusive. For example, the phrase "about 8" refers to values from 7.2 to 8.8, inclusive; as another example, the phrase "about 8%" refers to values from 7.2% to 8.8%, inclusive. Where present, all ranges are inclusive and can be combined. For example, when a range of "1 to 5" is recited, the recited range should be interpreted as including the ranges "1 to 4," "1 to 3," "1-2," "1-2 & 4-5," "1-3 & 5," and so on. In addition, when a list of alternatives is provided, such a list can also include embodiments that exclude any of the alternatives. For example, when describing a range of "1 to 5," such a description can support the case where any of 1, 2, 3, 4, or 5 is excluded; thus, a recited "1 to 5" can support "1 and 3-5, but not 2," or simply, "where 2 is not included."

As used herein, the terms "treatment" or "therapy" (and various word forms thereof) include prophylactic (e.g., preventative), therapeutic, or palliative treatment. Such prophylactic, therapeutic, or palliative treatment may be complete or partial. For example, complete elimination of an undesirable symptom or partial elimination of one or more undesirable symptoms represents "treatment" as contemplated herein.

As used above and throughout this disclosure, the term "effective amount" refers to an amount effective to achieve the desired result, at the necessary dosage and time, with respect to the treatment of the disorder, condition, or side effect. It should be understood that the effective amount of a component of the present invention will vary depending on the patient, not only depending on the specific compound, component, or composition selected, the route of administration, and the ability of the component to elicit the desired response in the individual, but also depending on factors such as the severity of the disease state or condition to be alleviated, the individual's hormone levels, age, sex, weight, the patient's condition and the severity of the condition being treated, concomitant medications or specific diet, as well as other factors considered by the specific patient and those skilled in the art. The appropriate dosage is ultimately determined by the attending physician. The dosage regimen can be adjusted to provide an improved therapeutic response. An effective amount is also an amount in which the beneficial effects of the treatment outweigh any toxic or adverse effects of the component. For example, a dose of a compound used in the method of the present invention is administered for a period of time such that platelet activation and adhesion activity are reduced compared to the activity levels before treatment begins.

"Pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that are, within the scope of sound medical judgment, suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problematic complications commensurate with a reasonable benefit/risk ratio.

Provided herein are crystalline forms of nicotinamide riboside chloride. While amorphous forms of nicotinamide riboside and its chloride salt are well known to those of ordinary skill in the art and have numerous uses, for example, derived from the ability of such molecules to enhance NAD production, the present disclosure relates to crystalline forms of these molecules. Crystalline forms of nicotinamide riboside possess advantageous properties, including chemical purity, fluidity, solubility, morphology or crystal habit, and stability (e.g., storage stability, dehydration stability, stability to polymorphic conversion, low hygroscopicity, and low levels of residual solvents).

The crystalline forms referred to herein are characterized by graphical data substantially as "described" in the accompanying drawings. Such data include, for example, powder X-ray diffraction patterns and solid-state IR spectra. A skilled artisan will appreciate that such graphical representations of the data may undergo minor variations, for example, in peak relative intensities and peak positions, due to factors such as variations in instrument response and variations in sample concentration and purity, as is well known to the skilled artisan. Nevertheless, a skilled artisan can readily compare the graphical data of the accompanying drawings herein with graphical data generated for an unknown crystalline form and determine whether the two sets of graphical data represent the same crystalline form or two different crystalline forms.

The present disclosure relates to crystalline forms of nicotinamide riboside, including Form I of nicotinamide riboside chloride according to Formula I

.

Crystalline Form I is characterized by a powder X-ray diffraction pattern having peaks at 5.1, 15.7, and 21.7 degrees 2θ ± 0.2 degrees 2θ. Crystalline Form I may also or alternatively be characterized by a powder X-ray diffraction pattern having peaks at 5.1, 15.7, 21.7, 23.5, and 26.4 degrees 2θ ± 0.2 degrees 2θ. Crystalline Form I may also or alternatively be characterized by a powder X-ray diffraction pattern having peaks at 5.1, 15.7, 18.6, 21.7, 23.5, 26.4, and 28.0 degrees 2θ ± 0.2 degrees 2θ.

In other embodiments, crystalline Form I is characterized by a powder X-ray diffraction pattern substantially as shown in Figure 1. Crystalline Form I may also or alternatively be characterized by a powder X-ray diffraction pattern having peaks substantially as provided in Table 1 ± 0.2 degrees 2Θ below.

Table 1

Crystalline Form I of nicotinamide riboside chloride may also or alternatively be characterized by a solid state IR spectrum having peaks at 671.7, 1035.6, and 1061.8 cm ^-1 ± 0.2 cm ^-1 . Crystalline Form I of nicotinamide riboside chloride may also or alternatively be characterized by a solid state IR spectrum having peaks at 671.7, 1035.6, 1061.8, 1398.9, and 1649.3 cm ^-1 ± 0.2 cm- ¹ . In certain embodiments, crystalline Form I of nicotinamide riboside chloride is characterized by a solid state IR spectrum substantially as shown in Figure 2. In other embodiments, crystalline Form I of nicotinamide riboside chloride is characterized by a solid state IR spectrum having peaks substantially as provided in Table 2 ± 0.2 cm ^-1 below.

Table 2

Another embodiment is directed to crystalline Form I of nicotinamide riboside chloride having a DSC thermogram substantially as shown in Figure 4.

In another embodiment, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 10 K/min, comprising an endotherm with an onset temperature of 119° C. ± 2° C. In certain instances, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 10 K/min, comprising an endotherm with an onset temperature of 118.8° C. ± 2° C.

In other embodiments, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 1 K/min comprising an endotherm with an onset temperature of 104°C ± 2°C, a peak temperature of 108°C ± 2°C, or both.

In other embodiments, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 2 K/min comprising an endotherm with an onset temperature of 109°C ± 2°C, a peak temperature of 113°C ± 2°C, or both.

In other embodiments, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 5 K/min comprising an endotherm with an onset temperature of 114°C ± 2°C, a peak temperature of 118°C ± 2°C, or both.

In another embodiment, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 10 K/min comprising an endotherm with a peak temperature of 123°C ± 2°C.

In another embodiment, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 10 K/min comprising an endotherm with an onset temperature of 119°C ± 2°C, a peak temperature of 123°C ± 2°C, or both.

In other embodiments, the crystalline Form I of nicotinamide riboside chloride is characterized by a DSC thermogram obtained using a heating rate of 20 K/min comprising an endotherm with an onset temperature of 122°C ± 2°C, a peak temperature of 128°C ± 2°C, or both.

It is known that the onset temperature and peak temperature and energy value of DSC can vary, for example, due to the purity and sample size of the sample, and due to instrument parameters, in particular the temperature scan rate. Therefore, the DSC data provided cannot be used as absolute values. For a differential scanning calorimeter, one skilled in the art can set the instrument parameters so that data comparable to the data provided herein can be collected according to standard methods, for example, as described in Höhne, G. W. H. et al., (1996), Differential Scanning Calorimetry, Springer, Berlin.

One embodiment of the present invention is directed to crystalline Form I of nicotinamide riboside chloride having a TGA/SDTA thermogram substantially as shown in Figure 7A.

The present disclosure also provides crystalline Form I of nicotinamide riboside chloride, characterized by a TGA/SDTA thermogram comprising an endotherm at 116°C ± 2°C and a mass loss of approximately 0.4%. The present disclosure also provides crystalline Form I of nicotinamide riboside chloride, characterized by a TGA/SDTA thermogram comprising an endotherm at 116.3°C ± 2°C and a mass loss of 0.36%.

Also disclosed is crystalline Form I of nicotinamide riboside chloride characterized by a DVS mass profile substantially as shown in Figure 8 A. In another embodiment, crystalline Form I of nicotinamide riboside chloride is characterized by a DVS isotherm profile substantially as shown in Figure 8B.

In another aspect, the present disclosure provides crystalline Form I of nicotinamide riboside chloride characterized by a water vapor sorption isotherm showing a water uptake of no more than about 0.5 wt % at a relative humidity of up to 60%. In another embodiment, crystalline Form I of nicotinamide riboside chloride is characterized by a water vapor sorption isotherm showing a water uptake of no more than about 0.5 wt %, preferably no more than about 1.0 wt %, at a relative humidity of up to 70%.

Crystalline Form I of the nicotinamide riboside chloride of the present invention can be provided in one of several different morphologies. For example, the crystalline material can exist in a morphology having a bulk density of about 0.25 to about 0.4 g/mL, or in a morphology having a bulk density of about 0.40 to about 0.65 g/mL. The present disclosure also contemplates mixtures of at least two morphologies in any ratio. FIG3A depicts a scanning electron microscope (SEM) image of crystalline nicotinamide riboside chloride of the present invention having a bulk density of about 0.25 to about 0.4 g/mL, and FIG3B depicts a scanning electron microscope (SEM) image of crystalline nicotinamide riboside chloride of the present invention having a bulk density of about 0.40 to about 0.65 g/mL. The inventors have discovered that the morphology of crystalline nicotinamide chloride having a bulk density of about 0.25 to about 0.4 g/mL is more stable to degradation by oxygen or water absorption. This morphology also appears to provide a slightly higher purity product. Due to the purity, stability, and color variations of other forms (forms having a bulk density of about 0.40 to about 0.65 g/mL), crystalline nicotinamide chloride having a bulk density of about 0.25 to about 0.4 g/mL is preferred, at least in some cases, because it appears to produce a higher quality, more consistent product.

In some embodiments, the crystalline Form I of nicotinamide riboside chloride is at least partially hydrated, and in other embodiments, the crystalline Form I of nicotinamide riboside chloride is anhydrous.

The present disclosure also relates to pharmaceutical compositions containing crystalline Form I of nicotinamide riboside chloride. The pharmaceutical compositions may comprise crystalline Form I of nicotinamide riboside chloride according to any of the above embodiments and a pharmaceutically acceptable excipient. The pharmaceutical compositions should comprise a therapeutically effective amount of crystalline Form I of nicotinamide riboside chloride.

As used herein, the phrase "therapeutically effective amount" refers to that amount of an active compound that will elicit the biological or medical response in a tissue, system, animal, individual, or human that is being sought by a researcher, veterinarian, medical doctor, or other clinician, and includes one or more of the following:

(1) preventing a disease or condition; for example, preventing a disease, condition, or disorder in an individual who may be susceptible to the disease, condition, or disorder but who does not yet experience or display signs or symptoms of the disease;

(2) inhibiting a disease or condition; e.g., inhibiting a disease, condition, or disorder in a subject experiencing or displaying signs or symptoms of the disease, condition, or disorder (i.e., including inhibiting further development of signs and/or symptoms); and

(3) Amelioration of a disease or condition; e.g., improvement of a disease, condition, or disorder in a subject experiencing or displaying signs or symptoms of the disease, condition, or disorder (i.e., including reversal of signs and/or symptoms).

Compositions of the present invention can be formulated for any type of administration. For example, compositions can be formulated for oral, topical, parenteral, enteral or inhalation administration. Described crystalline form I can be formulated for clean administration, or with conventional pharmaceutical carriers, diluents or excipient combinations that can be liquid or solid. Applicable solid carriers, diluents or excipients can especially play the effect of adhesive, disintegrant, filler, lubricant, glidant, compression aid, processing aid, pigment, sweetener, preservative, suspension/dispersant, tablet disintegrating agent, encapsulating material, film former or coating, flavoring or printing ink. Any material used in preparing any dosage unit form is preferably pharmaceutically pure, and is nontoxic substantially in the amount used. In addition, crystalline form I can be incorporated into sustained-release formulations and preparations. Administration in this regard particularly includes administration by the following routes: intravenous, intramuscular, subcutaneous, intraocular, intrasynovial, transepithelial (including transdermal), ocular, sublingual and oral, topical administration including ophthalmic, dermal, ocular, rectal and nasal inhalation by insufflation, aerosol and rectal systemic administration.

In powders, the carrier, diluent, or excipient can be a finely divided solid mixed with the finely divided active ingredient. In tablets, the active ingredient is mixed with a carrier, diluent, or excipient having the necessary compression properties in suitable proportions and compressed into the desired shape and size. For oral therapeutic administration, the active compound can be combined with a carrier, diluent, or excipient and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, and the like. In such therapeutically useful compositions, the amount of the active compound or compounds is preferably such that a suitable dosage is obtained.

Liquid carriers, diluents or excipients can be used to prepare solutions, suspensions, emulsions, syrups, elixirs, and the like. The active ingredient of the present invention can be dissolved or suspended in a pharmaceutically acceptable liquid (e.g., water, an organic solvent, a mixture of the two) or a pharmaceutically acceptable oil or fat. The liquid carrier, excipient or diluent can contain other suitable pharmaceutical additives, for example, solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavorings, suspending agents, thickeners, colorants, viscosity modifiers, stabilizers or osmotic pressure regulators.

Suitable solid carriers, diluents, and excipients can include, for example, calcium phosphate, silicon dioxide, magnesium stearate, talc, sugar, lactose, dextrin, starch, gelatin, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, polyvinyl pyrrolidine, a low melting wax, ion exchange resins, croscarmellose carbon, acacia, pregelatinized starch, crospovidone, HPMC, povidone, titanium dioxide, polycrystalline cellulose, aluminum methahydroxide, agar, tragacanth, or mixtures thereof.

For example, for oral, topical or parenteral administration, suitable examples of liquid carriers, diluents and excipients include water (especially containing the above-mentioned additives, such as cellulose derivatives, preferably sodium carboxymethylcellulose solution), alcohols (including monohydric and polyhydric alcohols, such as glycols) and their derivatives and oils (such as fractionated coconut oil and peanut oil) or mixtures thereof.

For parenteral administration, carrier, diluent or excipient can also be oily ester, for example, ethyl oleate and isopropyl myristate. Also contemplated are sterile liquid carriers, diluents or excipients, which are used in the sterile liquid form compositions of parenteral administration. Solutions of the active compounds of free alkali or pharmaceutically acceptable salt forms can be suitably mixed with surfactants (for example, hydroxypropyl cellulose) in water. Dispersants can also be prepared in glycerol, liquid polyethylene glycol and mixtures thereof and in oil. Under common storage and use conditions, in order to prevent the growth of microorganisms, these preparations can contain preservatives.

Pharmaceutical forms suitable for injection include, for example, sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. Where applicable, the form is preferably sterile and fluid to provide easy injectability. It is preferably stable under the conditions of preparation and storage and is preferably resistant to contaminating microorganisms (e.g., bacteria and fungi). The carrier, diluent, or excipient can be a solvent or dispersion medium containing, for example, water, ethanol, a polyol (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Suitable fluidity can be maintained, for example, by using a coating (e.g., lecithin), by maintaining the desired particle size (in the case of a dispersant), and by using a surfactant. Protection against microorganisms can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it is preferred to include an isotonic agent, such as sugar or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating a pharmaceutically appropriate amount of crystalline Form I of nicotinamide riboside chloride in a suitable solvent along with the various other ingredients listed above, as desired, followed by filtered sterilization. Generally, dispersions can be prepared by incorporating the sterilized active ingredient into a sterile vehicle containing a basic dispersion medium and, as required, the other ingredients listed above. In the case of sterile powders for the preparation of sterile injectable solutions, preferred preparation methods may include vacuum drying and freeze-drying techniques, which provide a powder of one or more active ingredients plus any other desired ingredients from a previously sterile-filtered solution thereof.

Thus, an effective amount of crystalline Form I of nicotinamide riboside chloride can be administered by any conventional technique recognized in the medical arts. For example, the amount administered can be from about 50 mg to about 50,000 mg per day. In some embodiments, the amount administered can be about 250 mg/kg/day. Thus, the amount administered can be about 50 mg/day, about 100 mg/day, about 200 mg/day, about 250 mg/day, about 300 mg/day, about 500 mg/day, about 700 mg/day, about 800 mg/day, about 1000 mg/day, about 2000 mg/day, about 4000 mg/day, about 5000 mg/day, about 10,000 mg/day, about 20,000 mg/day, about 30,000 mg/day, about 40,000 mg/day, or about 50,000 mg/day.

Also disclosed are methods for preparing such pharmaceutical compositions, comprising combining any previously disclosed embodiment of crystalline Form I of nicotinamide riboside chloride with a pharmaceutically acceptable excipient. Any acceptable method for combining an active agent with a pharmaceutically acceptable excipient may be used in accordance with the methods of the present invention, and suitable combination techniques will be readily apparent to one of ordinary skill in the art. In some embodiments, the combining step may be as simple as adding the desired amount of crystalline Form I of nicotinamide riboside chloride to an existing substance, such as a liquid beverage or powdered beverage mix. In other embodiments, the combining step comprises any technique commonly used to mix an active agent with an excipient to prepare a pharmaceutical dosage form (e.g., a solid, semisolid, liquid, or form suitable for inhalation), a cosmetic product (e.g., a powder, cream, lotion, or emollient), or a food product (e.g., a solid, semisolid, or liquid).

In other aspects, the present disclosure relates to a method comprising administering crystalline Form I of nicotinamide riboside chloride to a subject. Crystalline Form I of nicotinamide riboside chloride can be administered by any of the routes described above in connection with the pharmaceutical compositions of the present invention. For example, crystalline Form I of nicotinamide riboside chloride can be administered orally, topically, parenterally, enterally, or by inhalation. Due to the excellent stability of the crystalline Form I of nicotinamide riboside chloride disclosed herein, the active agent can be used or otherwise prepared for any known route of administration, and any known route of administration can be used according to the methods of the present invention. Crystalline Form I of nicotinamide riboside chloride can be administered in combination with a pharmaceutically acceptable excipient.

The subject or patient to whom the therapeutic compound is administered as an effective treatment regimen for a disease or disorder is preferably a human, but can be any animal, including laboratory animals in the context of a clinical trial or screening or activity experiment. Thus, one of ordinary skill in the art can readily appreciate that the methods, compounds, and compositions of the present invention are particularly suitable for administration to any animal, particularly mammals, including but not limited to humans, domestic animals, such as feline or canine subjects, farm animals, such as but not limited to cattle, horses, goats, sheep, and porcine subjects, wild animals (whether in the wild or in a zoo), research animals, such as mice, rats, rabbits, goats, sheep, pigs, dogs, cats, etc., avian species, such as chickens, turkeys, songbirds, etc., i.e., for veterinary medical use.

The present disclosure also provides a method for preparing crystalline Form I of nicotinamide riboside chloride. The method may include the steps of forming a solution containing nicotinamide riboside chloride and a polar solvent having hydrogen bonding, and cooling the combination. In some embodiments, the polar solvent having hydrogen bonding may be a polar alcohol. Exemplary polar alcohols include methanol, 1-butanol, 2-butanol, tert-butanol, diethylene glycol, ethanol, ethylene glycol, glycerol, 1-propanol, and 2-propanol. The polar solvent having hydrogen bonding may have high water solubility. For example, the polar solvent having hydrogen bonding may be acetone, acetonitrile, diethylene glycol dimethyl ether (DME), 1,2-dimethoxyethane (DME), dimethylformamide (DMF), dimethyl sulfoxide (DMSO), dioxane, hexamethylphosphoramide (HMPA), hexamethylphosphoric triamide (HMPT), N-methyl-2-pyrrolidone (NMP), or pyridine. The polar solvent having hydrogen bonding may bind to water. The solution may additionally comprise a source of water.

In some embodiments, forming the solution comprises combining crude nicotinamide riboside chloride with a polar solvent having hydrogen bonding. In other embodiments, the solution is formed by preparing nicotinamide riboside chloride in situ in the presence of a polar solvent having hydrogen bonding.

After the solution containing nicotinamide riboside chloride and the polar solvent having hydrogen bonding is formed, the mixture can be cooled at a temperature of about 15° C., about 10° C., about 0° C., about −10° C., about −15° C., about −20° C., or about −25° C. The mixture can be cooled for about 12 hours, about 18 hours, about 20 hours, about 24 hours, about 30 hours, about 36 hours, or about 40 hours.

After the cooling step, the method can further include adding an anti-solvent to the cooled composition (which now contains some crystalline product). As used herein, an "anti-solvent" is any material that helps the crystalline product to separate out from the solution. An exemplary anti-solvent is methyl tert-butyl ether (MTBE). After adding the anti-solvent to the cooled composition, the reaction mixture can be cooled for an additional period of time. The additional cooling time can be about 4 hours, about 6 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, or about 24 hours, and the cooling temperature can be about 15°C, about 10°C, about 0°C, about -10°C, about -15°C, about -20°C, or about -25°C.

After an additional cooling period, the solid produced in the previous step can be filtered and/or washed, for example, with an anti-solvent (eg, MTBE).

Also disclosed is crystalline Form I of nicotinamide riboside chloride prepared according to the above methods. Crystalline Form I of nicotinamide riboside chloride can be prepared according to any embodiment of the methods disclosed herein for forming crystalline forms.

The following examples further define the present invention. It should be understood that these examples, which represent preferred embodiments of the present invention, are given by way of illustration only and should not be construed as limiting the appended claims. From the above discussion and these examples, one skilled in the art will be able to ascertain the principal features of the present invention and, without departing from its spirit and scope, will be able to make various changes and modifications to the present invention to adapt it to various uses and conditions.

Example

Synthesis of crude nicotinamide riboside chloride

Many routes for synthesizing crude nicotinamide riboside and its chloride salt have been disclosed. Any known route or any other acceptable route may be used to prepare amorphous forms of the relevant compounds. Exemplary pathways for the synthesis of nicotinamide riboside and its chloride salt are disclosed in the following publications: Jarman et al., J. Chem. Soc. (1969), (2), 199-203 (chloride); Yang et al., J. Med. Chem. 2007, 50, 6458-6461; U.S. Publication No. 2007/0117765; Franchetti et al., Bioorg Med Chem Lett. 2004 Sep 20;14(18):4655-8; Saunders PP et al., Cancer Res. 1989 Dec 1;49(23):6593-9; Dowden J et al., Nucleosides Nucleotides Nucleic Acids. 2005;24(5-7):513-8; Schlenk, F., Archives of Biochemistry (1943), 3, 93-103; Freyne et al., Carbohydr. Res., 78:235-242 (1980); Tanimori et al., Bioorg. Med. Chem. Lett., 12:1135-1137 (2002); WO2010/017374; Davies LC, Nucleosides & Nucleotides 14(3-5), 311-312 1995; Kam BL et al., Carbohydrate Research, 77(1979)275-280; Viscontini M et al., VolumenXXXIX, Fasciculus VI(1956)-No. 195, 1620-1631. The entire disclosure of each of the references listed above is incorporated herein by reference.

Nicotinamide riboside can be initially synthesized using an anion other than ^Cl- (e.g., triflate or trifluoromethanesulfonate). After synthesizing this alternative form of nicotinamide riboside, it can be replaced with a chloride anion or another anion with higher affinity by means of ion exchange chromatography, thereby "displacing" the initial ion. Those of ordinary skill in the art can readily understand how to perform ion exchange chromatography.

Alternatively, amorphous nicotinamide riboside chloride can be obtained from commercial sources.

Preparation of crystalline nicotinamide riboside chloride

A solution containing methanol and nicotinamide riboside chloride is formed. After the solution is formed, it is cooled to -10°C and maintained at that temperature. After an additional 12-24 hours, the product begins to crystallize. The rate at which crystallization occurs can be increased by, for example, seeding the solution using known techniques. After this period, the mixture is determined to be a slurry, and 3 parts (the volume can vary, for example, 1-5 parts, depending on the amount of methanol) of methyl tert-butyl ether are slowly added over 6-12 hours. MTBE acts as an antisolvent to precipitate most of the product from the solution. The reaction mixture is then maintained at -10°C for an additional 12 hours. The solids are then filtered and rinsed with MTBE.

The aforementioned reaction/cooling times are based on a plant output of several hundred kilograms. When the reaction is carried out on a smaller scale, much of this time can be reduced without significantly affecting the morphology and appearance.

Preparation of amorphous nicotinamide riboside chloride

Experiments were performed to determine a suitable method for preparing substantially pure samples of amorphous nicotinamide riboside chloride for comparative studies with the compound's crystalline Form I. In Table 3 below, QSA1, QSA2, and QSA3 were performed to determine the solvent systems for preparing the amorphous samples.

Table 3

experiment solvent XRPD QSA1 water Amorphous QSA2 Dioxane/water (2:1) Amorphous QSA3 Ethanol/water (2:1) Oil QSA4 water Amorphous QSA5 Dioxane/water (2:1) Amorphous SASH Dioxane/water (2:1) Amorphous

According to QSA1-QSA3, the compound (nicotinamide riboside chloride) was dissolved in the selected solvent system (see Table 3). The vial was exposed to liquid nitrogen and the frozen solution was freeze-dried under vacuum. The ethanol/water solvent system resulted in an oil, and this sample was not used further. QSA4, QSA5, and SASH represent scaled-up freeze-drying experiments. The amorphous material of NR-Cl obtained according to QSA5 and SASH (dioxane/water solvent system) was slightly easier to handle (less viscous) than the amorphous material obtained with water (QSA4). Accordingly, all experiments involving amorphous material were performed using the sample obtained with QSA5. However, SEM images were obtained using the sample obtained under the conditions described for SASH.

instrument

X-ray powder diffraction. X-ray powder diffraction information for crystalline nicotinamide riboside chloride was obtained using a PANalytical X-PertPRO multi-purpose diffractometer (model #PY3040). No special sample preparation was required.

SEM. Scanning electron microscopy images were obtained using a Hitachi FE-SEM model #S-4500. No special sample preparation was required.

Infrared Spectra. Fourier transform infrared (FTIR) spectra were obtained using a Spectrum One™ FTIR instrument with universal attenuated total reflectance (Perkin-Elmer, Inc., Waltham, MA).

Differential Scanning Calorimetry (DSC)

DSC analysis of crystalline Form I of nicotinamide riboside chloride and amorphous nicotinamide riboside chloride was performed using a differential scanning calorimeter (Model DSC822e, Mettler-Toledo GmbH, Switzerland). The melting point of crystalline Form I of nicotinamide riboside chloride was measured using different heating rates, and the results are shown in Table 4 below:

Table 4

Heating rate Endothermic peak (℃) 1 K/min 108.01 2 K/min 112.53 5 K/min 118.43 10 K/min 123.25 20 K/min 127.82

Figure 4 provides DSC thermograms of crystalline Form I of nicotinamide riboside chloride measured at each experimental heating rate. Figure 5 provides a DSC thermogram of a sample of crystalline Form I of nicotinamide riboside chloride heated at a rate of 10 K/min. Figure 6 provides a DSC thermogram of an amorphous sample. As expected, DSC analysis of the amorphous form of nicotinamide riboside chloride did not yield a melting point.

Thermogravimetric mass spectrometry analysis

The mass loss due to loss of solvent or water in the crystalline Form I and amorphous form of nicotinamide riboside chloride was determined by TGA/SDTA. The sample weight was monitored during heating in a thermogravimetric analysis/simultaneous differential thermal analysis (TGA/SDTA) instrument model 851e (Mettler-Toledo GmbH, Switzerland) to obtain the respective mass versus temperature curves.

The TGA/SDTA851e was temperature-calibrated using indium and aluminum. The sample was weighed into a 100µl aluminum crucible and sealed. The seal was pin-holed, and the crucible was heated in a TGA from 25°C to 300°C at a heating rate of 10°C min ^-1 . Dry nitrogen was used for purging. The gases evolved from the TGA sample were analyzed by an Omnistar GSD 301 T2 mass spectrometer (Pfeiffer Vacuum GmbH, Germany). The former is a quadrupole mass spectrometer that analyzes masses in the 0-200 amu range.

TGA/SDTA thermograms of crystalline Form I of nicotinamide riboside chloride and an amorphous form of nicotinamide riboside chloride are shown in Figures 7A and 7B, respectively. In Figure 7A, the SDTA measurement of crystalline Form I shows an endotherm at 116.3°C, and the TGA measurement shows a mass loss of 0.36%. These results conclude that the compound is not a solvate and contains a small amount of residual solvent. In Figure 7B, the SDTA measurement of the amorphous sample shows an exotherm at 118.6°C, and the TGA measurement shows a mass loss of 1.59%. These results conclude that the compound is not a solvate and contains some residual solvent.

Hygroscopicity/Dynamic Vapor Sorption (DVS)

Hygroscopic isotherms were collected for the crystalline Form I of nicotinamide riboside chloride and the amorphous form of nicotinamide riboside chloride on a DVS-1 system from Surface Measurement Systems UK Ltd. (London, UK). Sample sizes ranged from 9.7 to 14.3 mg of solid material. The relative humidity was varied from 40% RH to 0% RH following the initial drying step. Subsequently, the relative humidity was raised to 95% (absorption), reduced to 0% RH (desorption), and raised again to 95% RH (absorption). A weight balance was set for each step, with a hold time of 1 hour (10% relative humidity step).

The sample size of each crystalline Form I sample was 12.6568 mg, and the amorphous sample was 9.6799 mg. Figure 8A provides a DVS mass change graph of a sample of crystalline Form I of nicotinamide riboside chloride, and Figure 8B provides a DVS isotherm graph of a sample of crystalline Form I of nicotinamide riboside chloride. Figure 9A provides a DVS mass change graph of a sample of the amorphous form of nicotinamide riboside chloride, and Figure 9B provides a DVS isotherm graph of a sample of the amorphous form of nicotinamide riboside chloride.

Figure 10 provides a comparison of the absorption curve of crystalline Form I of nicotinamide riboside chloride with that of an amorphous sample. This comparison shows that while both forms absorb water, there is a significant difference in the rate of absorption between 0% and 60% relative humidity: crystalline Form I is less prone to absorption at lower relative humidities than the amorphous material. Even at 70% relative humidity, the weight of the crystalline sample does not increase by more than approximately 1.0%. Overall, these characteristics of crystalline Form I facilitate handling of the material and represent an ability to remain stable over a wider range of operating conditions (relative to the amorphous form).

Claims (27)

1. Crystallization form I of nicotinamide nucleoside chloride according to formula I , The powder X-ray diffraction pattern is characterized by peaks at 5.1, 15.7, 21.7, 23.5 and 26.4 degrees 2θ ± 0.2 degrees 2θ. 2. The crystalline form I according to claim 1, characterized by a powder X-ray diffraction pattern having peaks at 5.1, 15.7, 18.6, 21.7, 23.5, 26.4 and 28.0 degrees 2θ ± 0.2 degrees 2θ. 3. The crystalline form I according to claim 1, characterized by a powder X-ray diffraction pattern substantially as shown in FIG1. 4. The crystalline form I according to claim 1, characterized by a powder X-ray diffraction pattern having peaks substantially as shown in Table 1 ± 0.2 degrees 2θ: Table 1 . 5. The crystalline form I according to claim 1, characterized by an IR spectrum with peaks at 671.7, 1035.6 and 1061.8 ^cm⁻¹ ± 0.2 ^cm⁻¹ . 6. The crystalline form I according to claim 1, characterized by an IR spectrum substantially as shown in FIG2. 7. The crystalline form I according to claim 1, characterized by having an IR spectrum with peaks substantially as shown in Table 2 ± 0.2 ^cm⁻¹ : Table 2 . 8. The crystalline form I according to claim 1, further characterized in that it is substantially as shown in FIG4, a DSC thermal analysis diagram. 9. The crystalline form I according to claim 1, further characterized in that the DSC thermal analysis chart obtained using a heating rate of 10 K/min includes an endothermic phenomenon with an initial temperature of 119℃±2℃ and an endothermic phenomenon with a peak temperature of 123℃±2℃, or includes both. 10. The crystalline form I according to claim 1, further characterized in that the DSC thermal analysis chart obtained using a heating rate of 1 K/min includes an endothermic phenomenon with an initial temperature of 104 °C ± 2 °C and a peak temperature of 108 °C ± 2 °C, or both. 11. The crystalline form I according to claim 1, further characterized in that the DSC thermal analysis chart obtained using a heating rate of 2 K/min includes an endothermic phenomenon with an initial temperature of 109℃±2℃ and a peak temperature of 113℃±2℃, or both. 12. The crystalline form I according to claim 1, further characterized in that the DSC thermal analysis chart obtained using a heating rate of 5 K/min includes an endothermic phenomenon with an initial temperature of 114℃±2℃ and a peak temperature of 118℃±2℃, or both. 13. The crystalline form I according to claim 1, further characterized in that the DSC thermal analysis chart obtained using a heating rate of 20 K/min includes an endothermic phenomenon with an initial temperature of 122℃±2℃ and a peak temperature of 128℃±2℃, or both. 14. The crystalline form I according to claim 1, further characterized in that it is substantially as shown in FIG7A, a TGA/SDTA thermal analysis diagram. 15. The crystalline form I according to claim 1, further characterized by a TGA/SDTA thermal analysis diagram, which includes endothermic phenomena at 116°C ± 2°C and a mass loss of 0.4%. 16. The crystalline form I according to claim 1, further characterized in that it is substantially as shown in the DVS mass variation diagram of FIG8A. 17. The crystalline form I according to claim 1, further characterized by a DVS isotherm diagram substantially as shown in FIG8B. 18. The crystalline form I according to claim 1, further characterized by a water vapor absorption isotherm that shows a water absorption of no more than 0.5 wt% at a relative humidity of up to 60%. 19. The crystalline form I according to claim 1, further characterized by a water vapor absorption isotherm that shows a water absorption of no more than 1.0 wt% at a relative humidity of up to 70%. 20. The crystalline form I according to claim 1, wherein the crystalline form I is anhydrous. 21. A pharmaceutical composition comprising crystalline form I according to any one of the preceding claims and a pharmaceutically acceptable excipient. 22. A method for preparing a pharmaceutical composition, the method comprising combining the crystalline form I according to any one of claims 1-20 with a pharmaceutically acceptable excipient. 23. A method for preparing crystalline form I of the nicotinamide nucleoside chloride according to claim 1. The method includes: It forms a solution containing nicotinamide nucleoside chloride and a polar solvent with hydrogen bonds; and Cool the assembly. The polar solvent with hydrogen bonds is methanol. 24. The method of claim 23, wherein the solution is formed by in-situ preparation of nicotinamide nucleoside chloride in the presence of the polar solvent having hydrogen bonds. 25. The method of claim 23, wherein the solution is formed by combining nicotinamide nucleoside chloride with the polar solvent having hydrogen bonds. 26. The method according to any one of claims 23-25, wherein the solution contains nicotinamide nucleoside chloride, a polar solvent having hydrogen bonds, and water. 27. The crystalline form I of nicotinamide nucleoside chloride prepared by the method according to any one of claims 23-26.