Crystalline composite, method for producing same, and composition

Abstract

The purpose of the present invention is to provide a novel crystal complex of nicotinamide mononucleotide, which is stable and highly safe. The present invention relates to a crystalline complex of a nicotinamide mononucleotide and a phenylalanine compound, the phenylalanine compound being L-phenylalanine and having peaks at diffraction angles (2 [theta]) of 19.7 + / -0.2 DEG, 26.4 + / -0.2 DEG, and 32.8 + / -0.2 DEG in a powder X-ray diffraction pattern measured using Cu-K [alpha] rays as an X-ray source. Alternatively, the phenylalanine-based compound is aspartame and has peaks at diffraction angles (2 [theta]) of 7.0 + / -0.2 DEG, 10.7 + / -0.2 DEG, 14.4 + / -0.2 DEG, and 29.1 + / -0.2 DEG in a powder X-ray diffraction pattern measured using a Cu-K [alpha] ray as an X-ray source.

Description

TECHNICAL FIELD

[0001] The present application relates to a novel crystalline complex of nicotinamide mononucleotide and a method for producing the same, and a composition containing the same. BACKGROUND

[0002] Nicotinamide mononucleotide (NMN) is one of biochemical substances that naturally exist in living organisms, and has been attracting attention in recent years as a substance that can inhibit aging and extend healthy life span. Patent Literature 1 discloses an amino acid salt of nicotinamide mononucleotide, and Patent Literature 2 discloses a co-crystal of nicotinamide mononucleotide and isoniazid.

[0003] PRIOR ART DOCUMENTS

[0004] PATENT LITERATURE

[0005] Patent Literature 1: Japanese Patent Application Laid-Open No. 2021-524501

[0006] Patent Literature 2: Japanese Patent Application Laid-Open No. 2023-001065 SUMMARY

[0007] PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] The amino acid salt of nicotinamide mononucleotide disclosed in Patent Literature 1 has high hygroscopicity due to its crystal structure, and therefore has a problem in handling when used in foods, health foods, supplements, pharmaceuticals, and the like. The co-crystal of nicotinamide mononucleotide and isoniazid disclosed in Patent Literature 2 uses isoniazid, which is not included in the standards for pharmaceutical additives and the Codex Alimentarius and has no food experience, and therefore the safety cannot be ensured when used in foods, health foods, supplements, pharmaceuticals, and the like.

[0009] The present application has been made in view of the above-described circumstances, and an object thereof is to provide a novel crystalline complex of nicotinamide mononucleotide, which is stable without deliquescence and has high safety because a component having a food experience is used.

[0010] MEANS FOR SOLVING THE PROBLEMS

[0011] In order to solve the above-described problems, the present inventors and the like have repeated intensive studies, and as a result, it has been found that nicotinamide mononucleotide can form a crystalline complex only with a part of compounds among a large number of compounds, and the part of compounds are safe L-phenylalanine and aspartame, which have a food experience, and they can form a crystalline complex with good safety and stability, and thus the present application has been completed.

[0012] That is, the gist of the present application is as described below.

[0013] [1] A crystalline complex of nicotinamide mononucleotide and a phenylalanine-based compound,

[0014] the phenylalanine-based compound is L-phenylalanine, and has at least one characteristic selected from the following (i) to (iii) in a diffraction pattern of powder X-ray diffraction measured using Cu-Kα rays as an X-ray source:

[0015] the phenylalanine-based compound is aspartame, and has peaks at diffraction angles (2θ) of 7.0±0.2°, 10.7±0.2°, 14.4±0.2°, and 29.1±0.2° in a diffraction pattern of powder X-ray diffraction measured using Cu-Kα rays as an X-ray source.

[0016] [2] The crystalline complex according to [1], wherein,

[0017] In the crystalline complex, the molar ratio of nicotinamide mononucleotide to the phenylalanine-based compound (nicotinamide mononucleotide:phenylalanine-based compound) is 1:0.7 to 1:1.3.

[0018] [3] The crystalline complex according to [1] or [2], wherein,

[0019] the phenylalanine-based compound is L-phenylalanine, and has at least one characteristic selected from the following (i) to (iii) in a diffraction pattern of powder X-ray diffraction measured using Cu-Kα rays as an X-ray source:

[0020] (i) the peak intensity ratio (peak intensity at 26.4±0.2° / peak intensity at 19.7±0.2°) at diffraction angles (2θ) of 19.7±0.2° and 26.4±0.2° is 0.35 or more and 0.90 or less;

[0021] (ii) the peak intensity ratio (peak intensity at 32.8±0.2° / peak intensity at 19.7±0.2°) at diffraction angles (2θ) of 19.7±0.2° and 32.8±0.2° is 0.10 or more and 0.60 or less;

[0022] (iii) the peak intensity ratio (peak intensity at 32.8±0.2° / peak intensity at 26.4±0.2°) at diffraction angles (2θ) of 26.4±0.2° and 32.8±0.2° is 0.30 or more and 0.80 or less.

[0023] [4] The crystalline complex according to [1] or [2], wherein,

[0024] The phenylalanine compound is aspartame, and in the powder X-ray diffraction pattern determined using Cu-Kα rays as the X-ray source, it has at least one characteristic selected from (a) to (f) below:

[0025] (a) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 7.0±0.2° (peak intensity at 7.0±0.2° / peak intensity at 10.7±0.2°) is greater than 0.30 and less than 0.70;

[0026] (b) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 10.7±0.2°) is greater than 0.05 and less than 0.50;

[0027] (c) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 10.7±0.2°) is greater than 0.05 and less than 0.50;

[0028] (d) The ratio of peak intensity at diffraction angles (2θ) of 7.0±0.2° to 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 7.0±0.2°) is greater than 0.30 and less than 0.80;

[0029] (e) The ratio of peak intensity at diffraction angles (2θ) of 7.0±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 7.0±0.2°) is greater than 0.30 and less than 0.70;

[0030] (f) The ratio of peak intensity at diffraction angles (2θ) of 14.4±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 14.4±0.2°) is greater than 0.50 and less than 1.05.

[0031] [5] A composition comprising any one of [1] to [4] crystalline complexes.

[0032] [6] A manufacturing method, which is a method for manufacturing the crystalline complex described in any one of [1] to [4], the method comprising:

[0033] Crystalline complexes are precipitated from a mixed solution containing dissolved nicotinamide mononucleotide and phenylalanine compounds.

[0034] [7] According to the manufacturing method described in [6], a crystalline complex of nicotinamide mononucleotide and phenylalanine compounds is precipitated from a mixed solvent of water and a water-mixed organic solvent.

[0035] [8] According to the manufacturing method described in [6] or [7], wherein,

[0036] The molar ratio of nicotinamide mononucleotide to phenylalanine compounds in the mixed solution (nicotinamide mononucleotide: phenylalanine compounds) is 1:0.7 to 1:1.3.

[0037] [9] According to the manufacturing method described in [7], wherein,

[0038] The water-mixed organic solvent comprises at least one selected from monohydric alcohols, ketones, and nitriles.

[0039]

[10] According to the manufacturing method described in [7] or [9], wherein,

[0040] The mixing ratio of water and water-mixed organic solvent in the mixed solvent is: relative to 1 unit volume of water, the water-mixed organic solvent is more than 0.5 units volume and less than 50 units volume.

[0041] The effects of the invention

[0042] According to the present invention, a novel crystalline complex of nicotinamide mononucleotide with safety and excellent stability, and a method thereof can be provided. Attached Figure Description

[0043] Figure 1 This is the powder X-ray diffraction pattern of the crystals obtained in Example 1.

[0044] Figure 2 This is the powder X-ray diffraction pattern of the crystals obtained in Example 2.

[0045] Figure 3 This is the powder X-ray diffraction pattern of the crystals obtained in Example 3.

[0046] Figure 4 This is the powder X-ray diffraction pattern of the crystals obtained in Example 4. Detailed Implementation

[0047] The crystalline complex of nicotinamide mononucleotide of the present invention is a complex with a safe phenylalanine compound with edible experience, and can be characterized using powder X-ray diffraction (PXRD). Specifically, it is a crystalline complex of nicotinamide mononucleotide and L-phenylalanine with peaks at diffraction angles (2θ) of 19.7±0.2°, 26.4±0.2°, and 32.8±0.2° in Cu-Kα-based powder X-ray diffraction; or a crystalline complex of nicotinamide mononucleotide and aspartame (N-(L-α-aspartic acid)-L-phenylalanine 1-methyl ester) with peaks at diffraction angles (2θ) of 7.0±0.2°, 10.7±0.2°, 14.4±0.2°, and 29.1±0.2° in Cu-Kα-based powder X-ray diffraction.

[0048] Because the crystalline complex of the present invention is safe and has excellent stability, it is easy to process and is advantageous for use in food, health food, supplements, pharmaceuticals, etc.

[0049] It should be noted that in powder X-ray diffraction, variations in the intensity of diffraction peaks can occur depending on the pretreatment method and sample setup. This is referred to as orientation or selective orientation. Due to this effect, the intensity of the characteristic peak, and even its intensity ratio with other peaks, may change, and the powder X-ray diffraction pattern may appear different. However, if the position of the characteristic peak, i.e., the position of the diffraction angle (2θ), is the same, it is essentially within the scope of this invention even if the peak intensity changes. That is, in determining the identity of crystallization based on powder X-ray diffraction, the position of the diffraction angle (2θ) and the similarity of the overall pattern are important. Furthermore, for the diffraction angle (2θ) in powder X-ray diffraction, an acceptable error range is within ±0.2°. In cases where the influence of amorphous substances or the like causes baseline disturbance in the powder X-ray diffraction pattern, it is preferable to correct the baseline to eliminate the influence of amorphous substances or the like.

[0050] Considering the above-mentioned effects, for example, as a crystalline complex of the above-mentioned nicotinamide mononucleotide and L-phenylalanine, it is preferable to have at least one characteristic selected from the following in Cu-Kα ray-based powder X-ray diffraction:

[0051] (i) The ratio of peak intensity at diffraction angles (2θ) of 19.7±0.2° and 26.4±0.2° (peak intensity at 26.4±0.2° / peak intensity at 19.7±0.2°) is 0.35 or more and 0.90 or less, preferably 0.40 or more and 0.80 or less, and more preferably 0.45 or more and 0.70 or less;

[0052] (ii) The peak intensity ratio at diffraction angles (2θ) of 19.7±0.2° and 32.8±0.2° (peak intensity at 32.8±0.2° / peak intensity at 19.7±0.2°) is 0.10 or more and 0.60 or less, preferably 0.15 or more and 0.50 or less, more preferably 0.20 or more and 0.45 or less; and

[0053] (iii) The peak intensity ratio at diffraction angles (2θ) of 26.4±0.2° and 32.8±0.2° (peak intensity at 32.8±0.2° / peak intensity at 26.4±0.2°) is 0.30 or more and 0.80 or less, preferably 0.35 or more and 0.70 or less, more preferably 0.40 or more and 0.65 or less. More preferably, it has two features selected from (i) to (iii) above, and even more preferably, it has three features selected from (i) to (iii) above.

[0054] Furthermore, considering the aforementioned effects, for example, as a crystalline complex of the aforementioned nicotinamide mononucleotide and aspartame, it is preferable to have at least one characteristic selected from the following in Cu-Kα ray-based powder X-ray diffraction:

[0055] (a) The peak intensity ratio at diffraction angles (2θ) of 10.7±0.2° and 7.0±0.2° (peak intensity at 7.0±0.2° / peak intensity at 10.7±0.2°) is 0.30 or more and 0.70 or less, preferably 0.35 or more and 0.65 or less, more preferably 0.40 or more and 0.60 or less;

[0056] (b) The peak intensity ratio at diffraction angles (2θ) of 10.7±0.2° and 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 10.7±0.2°) is 0.05 or more and 0.50 or less, preferably 0.10 or more and 0.40 or less, and more preferably 0.15 or more and 0.35 or less;

[0057] (c) The peak intensity ratio at diffraction angles (2θ) of 10.7±0.2° and 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 10.7±0.2°) is 0.05 or more and 0.50 or less, preferably 0.10 or more and 0.40 or less, and more preferably 0.15 or more and 0.35 or less;

[0058] (d) The peak intensity ratio at diffraction angles (2θ) of 7.0±0.2° and 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 7.0±0.2°) is 0.30 or more and 0.80 or less, preferably 0.35 or more and 0.70 or less, and more preferably 0.40 or more and 0.65 or less;

[0059] (e) The peak intensity ratio at diffraction angles (2θ) of 7.0±0.2° and 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 7.0±0.2°) is 0.30 or more and 0.70 or less, preferably 0.35 or more and 0.65 or less, more preferably 0.40 or more and 0.60 or less; and

[0060] (f) The peak intensity ratio at diffraction angles (2θ) of 14.4±0.2° and 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 14.4±0.2°) is 0.50 or more and 1.05 or less, preferably 0.60 or more and 1.00 or less, more preferably 0.70 or more and 0.95 or less. More preferably, it has two or more features selected from (a) to (f) above, even more preferably, it has four or more features selected from (a) to (f) above, and even more preferably, it has six features selected from (a) to (f) above.

[0061] In the crystalline complex, the molar ratio of nicotinamide mononucleotide to phenylalanine-based compound (L-phenylalanine or aspartame) (nicotinamide mononucleotide: phenylalanine-based compound) is preferably 1:0.7 to 1:1.3, more preferably 1:0.75 to 1:1.2, and even more preferably 1:0.8 to 1:1.1. The molar ratio of nicotinamide mononucleotide to phenylalanine-based compound in the crystalline complex can be calculated by using high-performance liquid chromatography (HPLC) and determining the content of nicotinamide mononucleotide in the crystalline complex.

[0062] Because the crystalline composite of the present invention has a unique crystalline structure, it is stable to moisture, and preferably also stable to heat. Therefore, if the crystalline composite of the present invention is used as a raw material, it does not deliquesce during the manufacturing process and storage, thus exhibiting good processability. Furthermore, the crystalline structure is not easily altered during the manufacturing process and storage, allowing for the manufacture of products (food, health food, supplements, pharmaceuticals, etc.) that maintain the same quality.

[0063] The crystalline complex of nicotinamide mononucleotide of the present invention can be manufactured by the following method, which includes, for example, a step of preparing a mixed solution in which nicotinamide mononucleotide and a phenylalanine compound (L-phenylalanine or aspartame) are dissolved; and a step of precipitating the crystalline complex from the mixed solution.

[0064] The solvent used in the above-described mixed solution is preferably selected from at least one of water and water-mixable organic solvents. It can be a single solvent or a mixed solvent, but is preferably a mixed solvent. When it is a single solvent, water is preferred. When it is a mixed solvent, a mixture of water and water-mixable organic solvents is preferred. Water-mixable organic solvents refer to organic solvents that are miscible with water.

[0065] Examples of water-mixable organic solvents include: monohydric alcohols such as methanol, ethanol, n-propanol, isopropanol, and n-butanol; ketones such as acetone, methyl ethyl ketone, and methyl isobutyl ketone; and tetrahydrofuran, 1,4-dimethyl ethyl ketone, etc. Ethers such as alkanes; nitriles such as acetonitrile and propionitrile; amides such as N,N-dimethylformamide; sulfoxides such as dimethyl sulfoxide; etc. Any one of these can be used alone, or two or more can be used in combination. As an aqueous mixed organic solvent, from the viewpoint of obtaining a crystalline complex with the desired crystalline structure with high efficiency, monohydric alcohols, ketones, and nitriles are preferred; more preferably, monohydric alcohols with 1 to 5 carbon atoms, ketones with 3 to 5 carbon atoms, and nitriles with 2 to 5 carbon atoms are preferred; even more preferably, monohydric alcohols with 1 to 4 carbon atoms, ketones with 3 to 4 carbon atoms, and nitriles with 2 to 4 carbon atoms are preferred; and acetonitrile is even more preferred. When using two or more aqueous mixed organic solvents, the nitrile (preferably acetonitrile) in the total amount of the aqueous mixed organic solvent is preferably 50% by mass or more, more preferably 70% by mass or more, even more preferably 80% by mass or more, and as an upper limit, for example, 99.9% by mass.

[0066] When the solvent used in the mixed solution is a mixture of water and a water-mixed organic solvent, from the viewpoint that a crystalline complex with the desired crystalline structure can be obtained with high efficiency, the water-mixed organic solvent is preferably 0.5 or more, more preferably 0.7 or more, and even more preferably 0.9 or more, with no particular upper limit, and can be, for example, 50 or less, relative to 1 unit volume of water.

[0067] The amount (volume) of the solvent (preferably a mixture of water and acetonitrile in a volume ratio of 1:1) used in the mixed solution is preferably 1 to 50 times (ml / mmol) relative to the molar amount of nicotinamide mononucleotide, more preferably 2 to 40 times (ml / mmol), and even more preferably 2.5 to 30 times (ml / mmol), from the viewpoint that a crystalline complex with the desired crystalline structure can be obtained with high efficiency.

[0068] From the viewpoint that a crystalline complex with the desired crystalline structure can be obtained with high efficiency, the molar ratio (nicotinamide mononucleotide: phenylalanine compound) of nicotinamide mononucleotide to phenylalanine compound in the mixed solution is preferably 1:0.7 to 1:1.3, more preferably 1:0.75 to 1:1.2, and even more preferably 1:0.8 to 1:1.1.

[0069] Mixing during the preparation of mixed solutions can be carried out using known or conventional apparatus and conditions.

[0070] The mixing during the preparation of the mixed solution is preferably carried out at room temperature or under heating. As for the mixing temperature, from the viewpoint that a crystalline complex with the desired crystalline structure can be obtained with high efficiency, it is preferably 10~80°C, more preferably 15~70°C, and even more preferably 20~60°C.

[0071] As a step in preparing a mixed solution, for example,

[0072] (A) An aqueous mixed organic solvent may be added after the nicotinamide mononucleotide and phenylalanine compounds have been dissolved in water;

[0073] (B) Nicotinamide mononucleotide and phenylalanine compounds can also be added to a mixture of water and water-mixed organic solvents to form a supersaturated state.

[0074] As a preparation step (A), specifically, nicotinamide mononucleotide and a phenylalanine compound are first dissolved in water at a volume of 1 to 50 times (ml / mmol), preferably 5 to 30 times (ml / mmol), relative to the molar amount of nicotinamide mononucleotide. Then, a water-mixed organic solvent is preferably stably added to the solution and mixed. Stable addition means preferably by adding the solvent dropwise over a period of at least 10 minutes. The addition time of the water-mixed organic solvent is preferably 10 to 200 minutes, more preferably 20 to 180 minutes, and even more preferably 30 to 150 minutes.

[0075] As a preparation step (B), specifically, it is preferable to first add an amount of nicotinamide mononucleotide exceeding its saturation solubility to a mixed solvent of water and a water-mixed organic solvent, remove the dissolved residue after dissolution, and then add an amount of phenylalanine compound exceeding its saturation solubility to the solution, remove the dissolved residue after dissolution, and obtain a mixed solution as a saturated solution. Furthermore, it is more preferable to further add a mixed solution of nicotinamide mononucleotide and phenylalanine compound to the above-mentioned saturated solution. When further adding nicotinamide mononucleotide and phenylalanine compound to the saturated solution, the amount (molar amount) of nicotinamide mononucleotide added to the saturated solution relative to the volume of water in the mixed solvent is preferably 0.01 to 5 times (mmol / ml), more preferably 0.03 to 2 times (mmol / ml), and even more preferably 0.05 to 1 times (mmol / ml). When nicotinamide mononucleotide and phenylalanine compounds are further added to the saturated solution, the molar ratio of the amount of nicotinamide mononucleotide and phenylalanine compounds added to the saturated solution (nicotinamide mononucleotide: phenylalanine compounds) is preferably 1:0.7 to 1:1.3, more preferably 1:0.75 to 1:1.2, and even more preferably 1:0.8 to 1:1.1.

[0076] Within the range that does not hinder the formation of crystalline complexes, auxiliary components such as antioxidants, ultraviolet absorbers, and stabilizers may be added to the mixed solution as needed.

[0077] In the process of precipitating a crystalline complex from a mixed solution, when the solvent of the mixed solution is a single solvent, crystallization is performed by cooling the mixed solution. When the solvent of the mixed solution is water and an aqueous mixed organic solvent, crystallization can occur without cooling because the aqueous mixed organic solvent acts as a poor solvent. However, from the viewpoint of obtaining a crystalline complex with the desired crystalline structure with high efficiency, cooling the mixed solution is preferred.

[0078] The cooling temperature of the mixed solution in the precipitation process of the crystalline complex is preferably above 0°C and below 20°C, more preferably above 2°C and below 15°C, and even more preferably above 3°C and below 12°C. It should be noted that if the cooling temperature is set to a temperature below 0°C, which would cause the mixed solution to freeze, the crystalline structure of the crystalline complex cannot be made into the desired structure, and therefore is not preferred.

[0079] The cooling time of the mixed solution in the precipitation process of the crystalline complex is preferably 0.5 to 96 hours, more preferably 1 to 72 hours, and even more preferably 3 to 60 hours.

[0080] In the cooling of the mixed solution during the precipitation process of the crystalline complex, the mixed solution can be stirred by known or conventional apparatus and conditions, and preferably allowed to stand.

[0081] The precipitated crystalline complex can be separated from the solvent by solid-liquid separation. Examples of solid-liquid separation methods include natural filtration, reduced pressure filtration, pressurized filtration, and centrifugal filtration. The crystalline complex obtained by solid-liquid separation can then be further processed by washing, recrystallization, and drying using known or conventional apparatus and conditions.

[0082] Based on nicotinamide mononucleotide, the yield of the crystalline complex obtained by the manufacturing method of the present invention is preferably 5 mol% or more, more preferably 10 mol% or more, and there is no particular upper limit, which can be 100 mol%.

[0083] The crystalline complex of the present invention can be used as a composition further comprising other ingredients, depending on the application. The compositions of the present invention possess safety and excellent stability due to the presence of the crystalline complex.

[0084] Other components include, for example, pharmaceutical ingredients and food ingredients, which can be contained in the composition in known combinations and proportions.

[0085] This application claims the benefit of priority based on Japanese Patent Application No. 2023-107092, filed on June 29, 2023. The entire contents of the description of Japanese Patent Application No. 2023-107092, filed on June 29, 2023, are incorporated herein by reference.

[0086] Example

[0087] The present invention will be described in more detail below with examples, but the present invention is not limited to the examples below. Of course, appropriate changes can be made within the scope of the above / below, and these are all included in the technical scope of the present invention.

[0088] The structural identification of crystalline complexes was performed by determining the X-ray diffraction pattern of the sample using powder X-ray diffraction. The conditions for performing powder X-ray diffraction are as follows.

[0089] Device: Rigaku Mini Flex II

[0090] Using X-rays: Cu-Kα rays

[0091] Strength: 30kV, 15mA

[0092] Angle: 2θ = 2~60°

[0093] Scanning speed: 2° / minute

[0094] Diverging slit (DS): 1.25°

[0095] Anti-scattering slit (SS): 1.25°

[0096] Receiving slit (RS): 0.3mm

[0097] The content of nicotinamide mononucleotide was determined by high performance liquid chromatography (HPLC). The HPLC conditions are shown below.

[0098] Chromatographic column: COSMOSIL 3PBr 3.0 × 150 mm (manufactured by Nacalai)

[0099] Column temperature: 40°C

[0100] Mobile phase: 20mM ammonium formate / MeOH = 95 / 5

[0101] Flow rate: 0.4 mL / min

[0102] Detector: UV 260nm

[0103] Sample injection volume: 5 μL (in water)

[0104] (Example 1)

[0105] 6.7 g (20 mmol) of nicotinamide mononucleotide and 3.4 g (20 mmol) of L-phenylalanine were dissolved in 200 mL of water at 55°C. After adding 200 mL of acetonitrile dropwise over 100 minutes, the solution was cooled to 4°C at a rate of 3°C / hr. The resulting slurry was dried and solidified at 30°C under reduced pressure using a vacuum pump, washed with acetonitrile, filtered, and dried to obtain 1.23 g of white crystals. Analysis of the crystals by powder X-ray diffraction revealed characteristic peaks not observed in the raw materials (nicotinamide mononucleotide and L-phenylalanine) at 2θ (±0.2°) = 19.68°, 26.35°, and 32.70°, confirming the formation of a crystalline complex with a different crystalline structure from the raw materials. Figure 1 The powder X-ray diffraction pattern of the crystal is shown. Furthermore, the obtained crystalline complex maintained its crystalline structure without deliquescence after being placed in an indoor environment for 24 hours.

[0106] (Example 2)

[0107] 6.7 g (20 mmol) of nicotinamide mononucleotide and 3.4 g (20 mmol) of L-phenylalanine were dissolved in 200 mL of water at 55°C. After adding 200 mL of acetonitrile dropwise over 113 minutes, the mixture was cooled to 4°C at a rate of 3°C / hr. The resulting slurry was concentrated at 30°C under reduced pressure using a vacuum pump, with 200 mL of acetonitrile added once during the concentration process, followed by further concentration and drying. The slurry was then washed with acetonitrile, filtered, and dried to obtain 2.25 g of white crystals. The crystals were analyzed by powder X-ray diffraction, and the results were similar to those in Example 1, showing characteristic peaks not observed in the raw materials (nicotinamide mononucleotide and L-phenylalanine) at 2θ (±0.2°) = 19.79°, 26.49°, and 32.83°, confirming the reproducible formation of a crystalline complex with a different crystalline structure from the raw materials. Figure 2 The powder X-ray diffraction pattern of the crystal is shown. Furthermore, the obtained crystalline complex maintained its crystalline structure without deliquescence after being placed in an indoor environment for 24 hours.

[0108] (Example 3)

[0109] 6.7 g (20 mmol) of nicotinamide mononucleotide and 5.89 g (20 mmol) of aspartame were dissolved in 200 mL of water at 55°C. After adding 200 mL of acetonitrile dropwise over 100 minutes, the solution was cooled to 4°C at a rate of 3°C / hr. The resulting slurry was concentrated under reduced pressure at 30°C, filtered, and dried to obtain 3.10 g of white crystals. The crystals were analyzed by powder X-ray diffraction, and unique peaks not observed in the raw materials nicotinamide mononucleotide and aspartame were observed at 2θ (±0.2°) = 7.05°, 10.70°, 14.44°, and 29.10°, confirming the formation of a crystalline complex with a different crystalline structure from the raw materials. Figure 3 The powder X-ray diffraction pattern of the crystal is shown. The obtained crystalline complex remained non-hygroscopic and maintained its crystalline structure after being placed in an indoor environment for 24 hours.

[0110] (Example 4)

[0111] 4.0 g of nicotinamide mononucleotide (NMN) was fully dissolved in 50 mL of a 1:1 mixture of acetonitrile and water at 25°C. After filtering out the residue, 1.0 g of L-phenylalanine was added and dissolved at 25°C. 1.67 g (5 mmol) of NMN and 0.83 g (5 mmol) of L-phenylalanine were added to the liquid obtained after further filtration (hereinafter referred to as the saturated solution). The solution was cooled to 6°C at a rate of 3°C / hr and then maintained at the same temperature for 2 days with stirring. The filtered slurry was dried at 30°C under vacuum to obtain 2.16 g of white crystals. The crystals were analyzed by powder X-ray diffraction (PXRD). Specific peaks not observed in the raw materials NMN and L-phenylalanine were observed at 2θ (±0.2°) = 19.77°, 26.25°, and 32.95°, confirming the formation of a crystalline complex with a different crystalline structure from the raw materials. Figure 4 The powder X-ray diffraction pattern of the crystal is shown. The amount of nicotinamide mononucleotide (NMN) in the crystal was determined by HPLC to be 55.4 mol%, indicating the formation of a crystalline complex with a NMN:phenylalanine ratio of 1:0.8. Furthermore, the obtained crystalline complex maintained its crystalline structure without deliquescence after being placed at room temperature for 24 hours.

[0112] ((Thermal stability test))

[0113] The crystalline complex obtained in Example 4 and the nicotinamide mononucleotide powder used as a control were heated at 150°C for 1 hour. The content of nicotinamide mononucleotide before and after heating was determined by HPLC, and the thermal stability was examined based on the residual amount of nicotinamide mononucleotide after heating relative to before heating. The results showed that the content of nicotinamide mononucleotide in the nicotinamide mononucleotide powder decreased to 81 mol% due to thermal decomposition, while the residual amount of nicotinamide mononucleotide in the crystalline complex obtained in Example 4 was 90 mol% after heating. The thermal stability was improved by becoming a crystalline complex.

[0114] (Comparative Example 1)

[0115] 1 g (3 mmol) of nicotinamide mononucleotide and 0.5 g (3 mmol) of L-phenylalanine were added to a container containing 80 mL of water. The container was cooled to freeze the contents, and then connected to a freeze dryer for freeze-drying to obtain a solid. The obtained solid was analyzed by powder X-ray diffraction. The X-ray diffraction pattern was different from that of the crystals obtained in Examples 1, 2, and 4, and no peaks characteristic of the crystalline complex of the present invention were observed. In addition, only extremely weak diffraction peaks were observed, indicating that the solid had low crystallinity. The obtained solid was hygroscopic, absorbing moisture immediately after being placed in an indoor environment, and becoming sticky over time.

Claims

1. A crystalline complex, which is a crystalline complex of nicotinamide mononucleotide and phenylalanine compounds, The phenylalanine compound is L-phenylalanine, and in the powder X-ray diffraction pattern measured using Cu-Kα rays as the X-ray source, it has peaks at diffraction angles (2θ) of 19.7±0.2°, 26.4±0.2°, and 32.8±0.2°; or The phenylalanine compound is aspartame, and in the powder X-ray diffraction pattern determined using Cu-Kα rays as the X-ray source, it has peaks at diffraction angles (2θ) of 7.0±0.2°, 10.7±0.2°, 14.4±0.2°, and 29.1±0.2°.

2. The crystalline composite according to claim 1, wherein, In the crystalline complex, the molar ratio of nicotinamide mononucleotide to phenylalanine compound (nicotinamide mononucleotide: phenylalanine compound) is 1:0.7 to 1:1.

3.

3. The crystalline composite according to claim 1, wherein, The phenylalanine compound is L-phenylalanine, and in the powder X-ray diffraction pattern measured using Cu-Kα rays as the X-ray source, it has at least one characteristic selected from (i) to (iii) below: (i) The ratio of peak intensity at diffraction angles (2θ) of 19.7±0.2° to 26.4±0.2° (peak intensity at 26.4±0.2° / peak intensity at 19.7±0.2°) is greater than 0.35 and less than 0.90; (ii) The ratio of peak intensity at diffraction angles (2θ) of 19.7±0.2° to 32.8±0.2° (peak intensity at 32.8±0.2° / peak intensity at 19.7±0.2°) is greater than 0.10 and less than 0.60; (iii) The ratio of peak intensity at diffraction angles (2θ) of 26.4±0.2° to 32.8±0.2° (peak intensity at 32.8±0.2° / peak intensity at 26.4±0.2°) is greater than 0.30 and less than 0.

80.

4. The crystalline composite according to claim 1, wherein, The phenylalanine compound is aspartame, and in the powder X-ray diffraction pattern determined using Cu-Kα rays as the X-ray source, it has at least one characteristic selected from (a) to (f) below: (a) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 7.0±0.2° (peak intensity at 7.0±0.2° / peak intensity at 10.7±0.2°) is greater than 0.30 and less than 0.70; (b) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 10.7±0.2°) is greater than 0.05 and less than 0.50; (c) The ratio of peak intensity at diffraction angles (2θ) of 10.7±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 10.7±0.2°) is greater than 0.05 and less than 0.50; (d) The ratio of peak intensity at diffraction angles (2θ) of 7.0±0.2° to 14.4±0.2° (peak intensity at 14.4±0.2° / peak intensity at 7.0±0.2°) is greater than 0.30 and less than 0.80; (e) The ratio of peak intensity at diffraction angles (2θ) of 7.0±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 7.0±0.2°) is greater than 0.30 and less than 0.70; (f) The ratio of peak intensity at diffraction angles (2θ) of 14.4±0.2° to 29.1±0.2° (peak intensity at 29.1±0.2° / peak intensity at 14.4±0.2°) is greater than 0.50 and less than 1.

05.

5. A composition comprising the crystalline complex of claim 1 or 2.

6. A manufacturing method for the crystalline composite of claim 1 or 2, the method comprising: Crystalline complexes are precipitated from a mixed solution containing dissolved nicotinamide mononucleotide and phenylalanine compounds.

7. The manufacturing method according to claim 6, wherein a crystalline complex of nicotinamide mononucleotide and phenylalanine compounds is precipitated from a mixed solvent of water and a water-mixed organic solvent.

8. The manufacturing method according to claim 6, wherein, The molar ratio of nicotinamide mononucleotide to phenylalanine compounds in the mixed solution (nicotinamide mononucleotide: phenylalanine compounds) is 1:0.7 to 1:1.

3.

9. The manufacturing method according to claim 7, wherein, The water-mixed organic solvent comprises at least one selected from monohydric alcohols, ketones, and nitriles.

10. The manufacturing method according to claim 7, wherein, The mixing ratio of water and water-mixed organic solvent in the mixed solvent is: relative to 1 unit volume of water, the water-mixed organic solvent is more than 0.5 units volume and less than 50 units volume.

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