A nicotinamide mononucleotide-proline cocrystal and its composition

Abstract

This invention aims to solve the problems of poor flowability of existing nicotinamide mononucleotide (NMN) single material, leading to poor flowability, uneven mixing, and poor content uniformity in NMN formulation preparation. It provides a safe and highly flowable nicotinamide mononucleotide-proline co-crystal. This co-crystal exhibits diffraction peaks at 5.5±0.2° and 16.8±0.2° using Cu-Ka radiation and X-ray powder diffraction at 2θ angles. This co-crystal possesses good bulk density and flowability, allowing for direct tableting or capsule filling of the powder. The product weight variation meets pharmacopoeia requirements, and the process is simple, making it suitable for commercial production. Furthermore, proline, a natural amino acid and a component of collagen, is safer and more reliable as a co-crystal form than isoniazid (classified as a Group 3 carcinogen by the WHO), which is disclosed in existing technologies.

Description

Technical Field

[0001] This invention relates to the field of compound crystal technology, and in particular to nicotinamide mononucleotide-proline cocrystal and its composition. Background Technology

[0002] β-Nicotinamide mononucleotide (NMN) is a direct precursor of coenzyme I (NAD+). As an important intermediate in the NAD salvage pathway within biological cells, its level directly affects NAD concentration. Coenzyme I is a crucial coenzyme in human redox reactions, participating in thousands of physiological responses. Studies have shown that the level of coenzyme I in the human body declines with age. In vitro NMN supplementation is the ideal way to effectively increase the level of coenzyme I in the body, potentially preventing cardiovascular disease, delaying aging, and preventing Alzheimer's disease. However, due to the insufficient stability of NMN, products prepared directly from amorphous powder are prone to degradation and inactivation during production, storage, and transportation.

[0003] Patent CN108697722A discloses a crystalline form of β-nicotinamide mononucleotide (NMN), revealing two crystalline forms of NMN and finding that the crystalline form can improve product stability. However, when used in oral dosage forms, problems such as poor flowability, difficulty in handling, and uneven content still exist. How to improve the relevant performance indicators and stability of NMN dosage forms is a problem that needs to be solved.

[0004] Patent CN113292619B discloses a nicotinamide mononucleotide-isonicotinic acid cocrystal, which includes a cocrystal of NMN and isoniazid, which can improve the problem of poor flowability. However, isoniazid, as a drug for treating tuberculosis, is highly toxic and was classified as a Group 3 carcinogen (possibly carcinogenic) by the World Health Organization (WHO) in 2017. Its use in healthy individuals poses safety risks. If the only goal is to address the product's poor flowability, a safer alternative must be chosen.

[0005] Drug cocrystals are solid crystals formed by the bonding of the active pharmaceutical ingredient (API) and cocrystal forming compound (CCF) through hydrogen bonds or other non-covalent bonds. The 2020 edition of the Chinese Pharmacopoeia's guidelines on crystal forms include cocrystal drugs within the category of crystalline substances. Cocrystal drugs can improve the mechanical processing properties, solubility, permeability, bioavailability, and stability of the active ingredient, but do not affect its biological characteristics. Summary of the Invention

[0006] The purpose of this invention is to provide a nicotinamide mononucleotide-proline cocrystal to solve the problems of uneven NMN content in NMN preparations due to poor NMN flowability, and the safety hazards caused by the use of the toxic reagent isoniazid as a cocrystal forming agent in existing technologies.

[0007] The above-mentioned technical objective of the present invention is achieved through the following technical solution: a nicotinamide mononucleotide-proline cocrystal, which, when subjected to Cu-Ka radiation, exhibits diffraction peaks at 5.5±0.2° and 16.8±0.2° in X-ray powder diffraction at an angle of 2θ.

[0008] Drug cocrystals are solid crystals formed by the bonding of the active pharmaceutical ingredient (API) and cocrystal forming compound (CCF) through hydrogen bonds or other non-covalent bonds. The 2020 edition of the Chinese Pharmacopoeia's guidelines on crystal forms include cocrystal drugs within the category of crystalline substances. Cocrystal drugs can improve the mechanical processing properties, solubility, permeability, bioavailability, and stability of the active ingredient, but do not affect its biological characteristics. The NMN-proline cocrystal provided in this invention was detected using Cu-Kα radiation and X-ray powder diffraction. Diffraction peaks were observed at 2θ angles of 5.5±0.2° and 16.8±0.2°. Typical figures are shown in [Figure number missing]. Figure 2 , and NMN single-component crystal form ( Figure 3 X-ray powder diffraction comparison (consistent with the crystal-free form disclosed in patent CN108697722 B) revealed the formation of new characteristic peaks, which were significantly different from those disclosed in patent CN113292619B, indicating the attainment of a new crystal form. This invention, based on the structures of NMN and proline, forms intermolecular hydrogen bonds to prepare a cocrystal, greatly improving the flowability of NMN. This cocrystal can be used to prepare the final oral dosage form product. The molecular structure of the cocrystal formed by NMN and proline through intermolecular hydrogen bonds is as follows: Figure 3 As shown. Proline is a natural amino acid, and its metabolite, 4-hydroxyproline, is an important component of animal collagen. Therefore, as a co-crystal form, it is safer and more reliable than isoniazid (CN113292619B) disclosed in the existing patent, and is more suitable for long-term use by healthy individuals. Through extensive experimental research, the inventors developed the above-mentioned co-crystal morphology, which presents a new crystal form of NMN. This crystal has a higher bulk density than existing crystals, significantly improving the flowability of single NMN and facilitating formulation preparation. Furthermore, the use of proline as the co-crystal form is safer and more reliable than isoniazid used in existing technologies.

[0009] Preferably, Cu-Ka radiation is used, and X-ray powder diffraction in 2θ angles shows diffraction peaks at 5.5±0.2°, 16.8±0.2°, 20.0±0.2°, 22.5±0.2°, 25.0±0.2°, 28.2±0.2°, 34.1±0.2°, and 40.0±0.2°.

[0010] Preferably, the eutectic has the following characteristics: Figure 2 The X-ray powder diffraction pattern shown.

[0011] Preferably, the eutectic has the following characteristics: Figure 4 The differential scanning calorimetry (DSC) graph is shown.

[0012] A second object of the present invention is to provide a nicotinamide mononucleotide composition comprising the above-described nicotinamide mononucleotide-proline cocrystal.

[0013] A third object of the present invention is to provide a medicament in which the active ingredient comprises the aforementioned nicotinamide mononucleotide-proline cocrystal. This medicament may be a tablet or a capsule, wherein the active ingredient is the aforementioned NMN-proline cocrystal provided by the present invention or a combination thereof with other excipients.

[0014] In summary, the present invention has the following beneficial effects:

[0015] This invention selects the natural amino acid proline as the co-crystal form, which is safer and more reliable than isoniazid (classified as a Group 3 carcinogen by the WHO) disclosed in the prior art. The NMN and proline co-crystal has better fluidity and a significantly higher bulk density compared to the single crystal form disclosed in the prior art. This improves the problem of poor fluidity during the preparation of tablets or capsules, which leads to uneven mixing and large differences in content. By utilizing the co-crystal, the stability of NMN is also improved through the formation of intermolecular hydrogen bonds, and the possibility of NMN degradation is reduced. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the NMN-proline cocrystal structure;

[0017] Figure 2 This is the X-ray diffraction pattern of NMN eutectic powder;

[0018] Figure 3 This is the X-ray diffraction pattern of NMN single-component powder;

[0019] Figure 4 This is the DSC spectrum of the NMN eutectic. Detailed Implementation

[0020] The present invention will be further described in detail below with reference to the accompanying drawings.

[0021] This specific embodiment is merely an explanation of the present invention and is not intended to limit the invention. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they are within the scope of the claims of the present invention.

[0022] All materials and reagents used in the examples were commercially available. NMN is consistent with the anhydrous crystal form 1 disclosed in patent CN108137639B.

[0023] Example 1:

[0024] Preparation of NMN-proline cocrystal

[0025] Dissolve 90g NMN and 35g proline in 2.5L of water, maintain the temperature at 40-50℃, slowly add 2.5L of methanol, stir for 5-10min, cool down by 5-10℃ and stir to precipitate crystals, filter and dry to obtain the NMN-proline cocrystals provided by this invention.

[0026] The NMN-proline prepared above was subjected to powder X-ray diffraction using a Shimadzu XRD-7000 with Cu-Ka radiation at a wavelength of 1.54 Å, a divergence slit of 1°, an X-ray tube voltage of 45 kV, an X-ray tube current of 40 mA, and a scanning range of 3-50° (2θ). The powder sample was flattened and placed in a microsample tray before detection. The X-ray powder diffraction pattern of the nicotinamide mononucleotide-proline cocrystal provided by this invention is shown below. Figure 2 As shown, the peaks and intensities corresponding to the diffraction angle 2θ are shown in Table 1 below.

[0027] Table 1 X-ray diffraction data of NMN eutectic powder

[0028] Serial Number 2θ / ° Relative strength / % 1 5.5 100 2 16.7 36 3 20.0 8 4 22.5 16 5 25.0 5 6 28.2 10 7 34.1 7 8 40.0 13

[0029] Differential scanning calorimetry (DSC) was performed on the NMN-proline cocrystals prepared above.

[0030] DSC measurements were performed in a NETZSCH (DSC214) instrument using a sealed disk apparatus. Approximately 3 mg of sample was weighed into an aluminum disk, accurate to 0.1 mg. The instrument was purged with nitrogen at 50 ml / min, with a protective gas flow rate of 70 ml / min. The temperature was increased at a rate of 10 °C / min between room temperature and 220 °C. A graph was plotted with the absorption peak pointing downwards. The x-axis represents temperature (°C), and the y-axis represents the heat flux released per unit mass (Mw / mg). See the DSC graph below. Figure 4 .

[0031] Example 2

[0032] Bulk density determination

[0033] Take appropriate amounts of NMN single component (anhydrous crystals) and NMN-proline cocrystals obtained in Example 1 above, sieve them, weigh them accurately, and follow the general principles of the 2020 edition of the Chinese Pharmacopoeia. <0993> Bulk density and tap density were determined by the first method, the solid mass method. The bulk density was examined and the results are shown in Table 2 below. It can be seen that the bulk density of the eutectic is significantly higher than that of the single-component NMN, that is, the fluidity is improved. Considering the measurement error, the bulk density is consistent with that of the isoniazid eutectic disclosed in the embodiment of patent CN113292619B (0.68 g / ml).

[0034] Table 2 Results of Bulk Density Study

[0035] crystal Bulk density g / ml NMN single component (anhydrous crystals) 0.18 Example 1 Eutectic 0.65

[0036] Example 3

[0037] Content uniformity determination

[0038] Take NMN single component (anhydrous crystals) and NMN-proline cocrystal obtained in Example 1 above, respectively, pass them through a 200-mesh sieve, and fill them into capsules using #2 capsule shells according to the labeled amount of 0.2g, in accordance with the 2020 edition of the Chinese Pharmacopoeia General Rules. <0103> The fill weight difference was measured using the method described in Table 3 below. It can be seen that the fill weight difference of the eutectic obtained by the present invention is small, and when used to prepare tablets or capsules, there will be no problem of uneven NMN content or large quality difference.

[0039] Table 3 Results of the investigation on dosage variation

[0040] crystal Content variation / % in conclusion Filling volume variation limit NMN single component (anhydrous crystals) 30.2 Does not meet requirements ±10% Example 1 Eutectic 6.7 Meets requirements

Claims

1. A nicotinamide mononucleotide-proline co-crystal, characterized in that, X-ray powder diffraction, expressed in terms of 2 theta angle, using Cu-Ka radiation, has diffraction peaks at 5.5±0.2°, 16.8±0.2°, 20.0±0.2°, 22.5±0.2°, 25.0±0.2°, 28.2±0.2°, 34.1±0.2°, 40.0±0.2°.

2. The nicotinamide mononucleotide-proline co-crystal of claim 1, wherein, The co-crystal has an X-ray powder diffraction pattern as shown in Figure 2.

3. The nicotinamide mononucleotide-proline co-crystal of claim 1, wherein, The co-crystal has a differential scanning calorimetry profile as shown in Figure 4.

4. A nicotinamide mononucleotide composition, characterized in that, The composition comprises the nicotinamide mononucleotide-proline co-crystal according to any one of claims 1 to 3.

5. A medicament, characterized by comprising: The active ingredient of the medicament comprises the nicotinamide mononucleotide-proline co-crystal according to any one of claims 1 to 3.

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