A preparation process of nicotinamide mononucleotide

Abstract

The present invention provides a preparation process of nicotinamide mononucleotide, the steps include: dissolving nicotinamide riboside in 2-methyltetrahydrofuran, adding phosphorus pentoxide for phosphorylation reaction; after completion, heat preservation and stirring, then gradient temperature increase, stirring to obtain a solution of the product; slowly adding sodium hydroxide aqueous solution to the solution of the product to the system, stirring and dissolving, standing and stratifying; decompressing and concentrating the water layer obtained by stratification to half of the total volume, collecting the water obtained by decompression and concentration, cooling and stirring to crystallize, filtering and removing phosphate to obtain a nicotinamide mononucleotide suspension solution; adding ethanol for refining, crystallizing and cooling, and filtering to obtain a nicotinamide mononucleotide crude product; finally, the crude product is refined again in a methanol aqueous solution, crystallized and cooled, filtered, and decompressed and dried to obtain a nicotinamide mononucleotide product. A preparation process of nicotinamide mononucleotide proposed by the present invention has a high yield, and the product has high purity and few impurities, and can also reduce the residual heavy metal arsenic in the product.

Description

Technical Field

[0001] The present invention relates to the technical field of organic synthesis, and in particular to a preparation process of nicotinamide mononucleotide. Background Art

[0002] Nicotinamide mononucleotide (NMN), as a naturally occurring bioactive nucleotide, has a unique chemical structure consisting of a phosphate group, a ribose molecule, and a nicotinamide base connected by specific chemical bonds. This structure gives NMN unique biological activity and a wide range of physiological functions. As the human body ages, the level of NAD+ (nicotinamide adenine dinucleotide) in cells gradually decreases. As an important coenzyme for key biological processes such as intracellular energy metabolism and DNA repair, the decrease in NAD+ levels is closely related to the occurrence of a variety of aging-related diseases. Therefore, people have begun to pay attention to and hope to increase the level of NAD+ in the body by supplementing NMN, thereby delaying the aging process and preventing related diseases.

[0003] In recent years, with the continuous deepening of research on NMN, scientists have gradually revealed its significant role in delaying aging and preventing neurodegenerative diseases. These findings have not only attracted widespread attention in the academic community, but also stimulated consumers' strong interest in NMN products. The research enthusiasm for NMN continues to rise at home and abroad, and more and more scientific research institutions and companies have invested in the research and development and production of NMN, which has promoted the rapid development of the NMN market.

[0004] As the safety and effectiveness of NMN have been more comprehensively recognized and affirmed, its application fields and scope of application are also expanding. At present, NMN has been widely used in many fields such as health products, cosmetics, and biomedicine, providing new solutions for human health and beauty. However, despite the growing market demand for NMN, its preparation process still faces many challenges. How to prepare high-purity NMN efficiently and environmentally friendly has become a hot and difficult issue in current research. Based on this, the present invention provides a preparation process of nicotinamide mononucleotide. Summary of the invention

[0005] The purpose of the present invention is to provide a preparation process of nicotinamide mononucleotide, which can effectively reduce the residual heavy metal arsenic in the product.

[0006] The present invention provides a process for preparing nicotinamide mononucleotide, the steps comprising: (1) Dissolve nicotinamide riboside in 2-methyltetrahydrofuran, and add phosphorus pentoxide in batches for phosphorylation reaction; after the addition is completed, keep warm and stir, then gradually increase the temperature to 10°C-15°C, and stir for 5-7 hours to obtain a solution of the product; (2) Slowly add 10% sodium hydroxide aqueous solution into the solution of this product, stir to dissolve, and let stand to separate into layers; (3) Concentrating the aqueous layer obtained by stratification under reduced pressure to half of the total volume, collecting the water obtained by reduced pressure concentration, cooling and stirring to crystallize, filtering to remove phosphate, and obtaining a nicotinamide mononucleotide suspension solution; (4) adding ethanol for refining, crystallizing, cooling, and filtering to obtain a crude nicotinamide mononucleotide; (5) Finally, the crude product is purified again in a methanol aqueous solution, crystallized, cooled, filtered, and dried under reduced pressure to obtain the nicotinamide mononucleotide product.

[0007] Furthermore, in step (1), the weight ratio of nicotinamide riboside, 2-methyltetrahydrofuran and phosphorus pentoxide is (8-12): (180-220): (35-45).

[0008] Furthermore, the temperature of the heat-insulating stirring in step (1) is 0°C-10°C.

[0009] Furthermore, the heating rate of the gradient heating is 1°C every 5-10 minutes.

[0010] Furthermore, in step (3), the temperature after cooling is -5°C, and the stirring and crystallization time is 10-14h.

[0011] Furthermore, in the step (4), the crystallization cooling is performed by stirring at 10°C-15°C for 28-32 minutes, then cooling to -5°C and keeping warm for 55-65 minutes.

[0012] Furthermore, the filtration in step (4) is vacuum filtration, and is performed until there are no droplets left.

[0013] Furthermore, in the step (5), the weight ratio of methanol to water in the methanol aqueous solution is (14-16): (1-3).

[0014] Furthermore, in the step (5), the crystallization cooling is performed by stirring at 10°C-15°C for 28-32 minutes, then cooling to -5°C and keeping warm for 55-65 minutes; and the filtration is performed by vacuum filtration until there is no droplet at all.

[0015] Furthermore, in step (5), the temperature of the reduced pressure drying is 25°C-30°C and the vacuum degree is -0.09 MPa.

[0016] The beneficial effects of the present invention are: The method of the present invention uses NR (nicotinamide riboside) as a starting material, performs a phosphorylation reaction with phosphorus pentoxide to obtain an NMN solution, neutralizes excess phosphorus pentoxide with a 10% sodium hydroxide aqueous solution, removes phosphate by filtration, and removes the organic layer by layer to obtain an NMN aqueous solution; the NMN aqueous solution is concentrated under reduced pressure to half the total volume, ethanol is added to the system for stirring and crystallization, the temperature is lowered, and the crude NMN product is filtered to obtain a crude NMN product, which is then refined with a methanol / water mixed solvent, cooled, filtered, and dried under reduced pressure to a constant weight to obtain an NMN product. The method of the present invention can better control the residual heavy metal arsenic in the product.

[0017] The present invention adopts phosphorus pentoxide to replace traditional phosphorus oxychloride and trimethyl phosphate for phosphorylation reaction. This improvement significantly reduces the residual amount of heavy metal arsenic in the final product, thereby improving the safety of the product and ensuring the improvement of product quality.

[0018] The present invention effectively removes most of the phosphates by implementing the filtration step, thereby reducing the residual phosphorus-containing compounds in the water environment. This measure significantly improves the environmental performance of the product, reduces the potential impact on the environment, and makes the product more in line with environmental protection standards.

[0019] The present invention abandons the use of phosphorus oxychloride, and reduces potential safety hazards in the actual operation process. By avoiding the use of this highly dangerous chemical, not only the safety of the operator is improved, but also the potential risk of environmental pollution is reduced, making the entire production process safer and more reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 It is a preparation process flow chart of the present invention; Figure 2 is a liquid phase purity spectrum of nicotinamide mononucleotide prepared in Example 1; Figure 3 is the liquid phase purity spectrum of nicotinamide mononucleotide prepared in Example 2; Figure 4 This is the liquid phase purity spectrum of nicotinamide mononucleotide prepared in Example 3. DETAILED DESCRIPTION

[0022] The technical scheme of the present invention is described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without creative work are within the scope of protection of the present invention. It should be noted that the raw materials can be obtained commercially.

[0023] The advantage of this preparation method is that it can better control the residual heavy metal arsenic in the product.

[0024] like Figure 1 As shown, a process for preparing nicotinamide mononucleotide of the present invention comprises the following steps: (1) Dissolve 8-12g of nicotinamide riboside in 180-220g of 2-methyltetrahydrofuran, control the system temperature at 0℃-10℃, and add 35-45g of phosphorus pentoxide to the solution in batches; after the addition, keep the temperature at 0℃-10℃ and stir for 55-65min; then increase the temperature gradually, increase the temperature by 1℃ every 5min-10min to 10℃-15℃, and stir evenly for 5-7h to obtain a solution of this product; (2) Slowly add 10% sodium hydroxide aqueous solution to the solution of this product, stir and dissolve it, let it stand for 30 minutes, separate the layers, and obtain the lower aqueous layer (i.e., the material layer), and discard the upper organic phase; (3) The water layer (material layer) obtained by stratification is concentrated to half of the total volume, and the vacuum concentration is completed. The water obtained by the vacuum concentration is collected, and the system continues to stir to reduce the internal temperature to -5°C, and stir to crystallize for 10-14 hours, and filter to remove the phosphate to obtain a nicotinamide mononucleotide suspension solution; (4) Add 14-16 g of ethanol to the nicotinamide mononucleotide suspension solution, stir at 10°C-15°C for 28-32 min, then cool to -5°C and keep warm for 55-65 min, and vacuum filter until there are no droplets to obtain a crude wet product of nicotinamide mononucleotide; (5) The crude wet product of nicotinamide mononucleotide is then added to a mixture of methanol and water in a weight ratio of (14-16): (1-3), stirred at 10°C-15°C for 28-32 minutes, then cooled to -5°C and kept warm for 55-65 minutes, and vacuum filtered until there are no droplets, thereby obtaining a wet product of nicotinamide mononucleotide; the wet product of nicotinamide mononucleotide is dried under reduced pressure at 25°C-30°C and a vacuum degree of -0.09 MPa until it reaches a constant weight on an infrared dryer, which is considered to be the end of drying, and a white dry product is finally obtained.

[0025] The specific reactions are as follows:

[0026] An appropriate amount of 10% sodium hydroxide aqueous solution is used to neutralize the excess phosphorus pentoxide in the system to generate sodium phosphate.

[0027] The present invention will be further explained below with examples: Example 1

[0028] 10g of nicotinamide riboside was dissolved in 200g of 2-methyltetrahydrofuran, the system temperature was controlled at 5°C, and 40g of phosphorus pentoxide was added to the solution in batches; after the addition, the mixture was kept at 5°C and stirred for 1h; then the temperature was increased gradually, increasing by 1°C to 12°C every 7min, and stirred evenly for 6h to obtain the solution of this product; a 10% sodium hydroxide aqueous solution was slowly added to the solution of this product, stirred and dissolved, and allowed to stand for 30min, and the layers were separated to obtain the lower water layer (i.e., the material layer), and the upper organic phase was discarded; the water layer (material layer) obtained by separation was concentrated to half of the total volume, and the reduced pressure concentration was completed, and the water obtained by the reduced pressure concentration was collected, and the system continued to stir to reduce the internal temperature to -5°C, stirred and crystallized for 12h, and filtered to remove the phosphate to obtain NMN Suspension solution; add 15g ethanol to the NMN suspension solution, stir at 12℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain NMN crude wet product; then add the NMN crude wet product to a methanol / water mixture (a mixture of 15g methanol and 2g purified water), stir at 12℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain NMN wet product; the NMN wet product is dried under reduced pressure at 27℃ and a vacuum degree of -0.09MPa to infrared dry weight until constant weight is reached, and the drying is considered to be completed. Finally, 6.2g of off-white dry product is obtained with a purity of 99.907% and a heavy metal arsenic residue of 1.1ppm.

[0029] The liquid phase purity spectrum of the product prepared in this example is as follows Figure 2 The corresponding data is shown in the following table: Example 2

[0030] 10g of nicotinamide riboside was dissolved in 200g of 2-methyltetrahydrofuran, the system temperature was controlled at 5°C, and 50g of phosphorus pentoxide was added to the solution in batches; after the addition, the mixture was kept at 5°C and stirred for 1h; then the temperature was increased gradually, increasing by 1°C to 12°C every 7min, and stirred evenly for 6h to obtain the solution of this product; a 10% sodium hydroxide aqueous solution was slowly added to the solution of this product, stirred and dissolved, and allowed to stand for 30min, and the layers were separated to obtain the lower water layer (i.e., the material layer), and the upper organic phase was discarded; the water layer (material layer) obtained by separation was concentrated to half of the total volume, and the reduced pressure concentration was completed, and the water obtained by the reduced pressure concentration was collected, and the system continued to stir to reduce the internal temperature to -5°C, stirred and crystallized for 12h, and filtered to remove the phosphate to obtain NMN Suspension solution; add 15g ethanol to the NMN suspension solution, stir at 12℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain NMN crude wet product; then add the NMN crude wet product to a methanol / water mixture (a mixture of 15g methanol and 2g purified water), stir at 12℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain NMN wet product; the NMN wet product is dried under reduced pressure at 27℃ and a vacuum degree of -0.09MPa to infrared dry weight until constant weight is reached, and the drying is considered to be complete, obtaining 6.2g of off-white dry product with a purity of 99.892% and 1.1ppm of heavy metal arsenic residue.

[0031] The liquid phase purity spectrum of the product prepared in this example is as follows Figure 3 The corresponding data is shown in the following table: Example 3

[0032] Dissolve 10g of nicotinamide riboside in 200g of 2-methyltetrahydrofuran, control the system temperature at 10°C, and add 60g of phosphorus pentoxide to the solution in batches; after the addition, keep the temperature at 10°C and stir for 1h; then increase the temperature gradually, increase the temperature by 1°C to 15°C every 5min, and stir evenly for 6h to obtain the solution of this product; slowly add a 10% sodium hydroxide aqueous solution to the solution of this product into the system and stir to dissolve, let it stand for 30min, separate the layers, obtain the lower water layer (i.e., the material layer), and discard the upper organic phase; concentrate the water layer (material layer) obtained by separation to half of the total volume, and end the vacuum concentration, collect the water obtained by the vacuum concentration, continue to stir the system to reduce the internal temperature to -5°C, stir and crystallize for 12h, filter to remove the phosphate, and obtain NMN Suspension solution; add 15g of ethanol to the NMN suspension solution, stir at 10℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain a crude NMN wet product; then add the crude NMN wet product to a methanol / water mixture (a mixture of 15g methanol and 2g purified water), stir at 10℃ for 30min, then cool to -5℃, keep warm for 1h, and vacuum filter until there are no droplets to obtain a wet NMN product; the wet NMN product is dried under reduced pressure at 25℃ and a vacuum degree of -0.09MPa and dried by infrared drying to constant weight, which is considered to be the end of drying, and 6.5g of off-white to light yellow dry product is obtained with a purity of 99.777% and a heavy metal arsenic residue of 1.5ppm.

[0033] The liquid phase purity spectrum of the product prepared in this example is as follows Figure 4 The corresponding data is shown in the following table:

[0034] The applicant declares that the present invention illustrates the product and detailed preparation method of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned product and detailed preparation method, that is, it does not mean that the present invention must rely on the above-mentioned product and detailed preparation method to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of various raw materials of the product of the present invention and addition of auxiliary materials, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0035] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited to the specific details in the above embodiments, and the technical solution of the present invention can be subjected to a variety of simple modifications within the technical concept of the present invention. These simple modifications all belong to the protection scope of the present invention.

[0036] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

[0037] In addition, various embodiments of the present invention may be arbitrarily combined, and as long as they do not violate the concept of the present invention, they should also be regarded as the contents disclosed by the present invention.

Claims

1. A process for preparing nicotinamide mononucleotide, characterized in that the steps include: (1) dissolving nicotinamide riboside in 2-methyltetrahydrofuran, and adding phosphorus pentoxide in batches for phosphorylation reaction; After the addition is completed, the mixture is stirred at a constant temperature, and then the temperature is gradually increased to 10°C-15°C, and stirred for 5-7 hours to obtain a solution of the product; (2) Slowly add 10% sodium hydroxide aqueous solution into the solution of this product, stir to dissolve, and let stand to separate into layers; (3) Concentrating the aqueous layer obtained by stratification under reduced pressure to half of the total volume, collecting the water obtained by reduced pressure concentration, cooling and stirring to crystallize, filtering to remove phosphate, and obtaining a nicotinamide mononucleotide suspension solution; (4) adding ethanol for refining, crystallizing, cooling, and filtering to obtain a crude nicotinamide mononucleotide; (5) Finally, the crude product is purified again in a methanol aqueous solution, crystallized, cooled, filtered, and dried under reduced pressure to obtain the nicotinamide mononucleotide product.

2. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: In the step (1), the weight ratio of nicotinamide riboside, 2-methyltetrahydrofuran and phosphorus pentoxide is (8-12): (180-220): (35-45).

3. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: The temperature of the heat preservation and stirring in step (1) is 0°C-10°C.

4. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: The heating rate of the gradient heating is 1°C every 5-10 minutes.

5. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: The temperature after cooling in step (3) is -5°C, and the stirring crystallization time is 10-14h.

6. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: In the step (4), the crystallization is cooled by stirring at 10°C-15°C for 28-32 minutes, then cooling to -5°C and keeping warm for 55-65 minutes.

7. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: The filtration in step (4) is vacuum filtration, and the filtration is continued until there are no droplets left.

8. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: In the step (5), the weight ratio of methanol to water in the methanol aqueous solution is (14-16): (1-3).

9. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: In the step (5), the crystallization cooling is performed by stirring at 10°C-15°C for 28-32 minutes, then cooling to -5°C and keeping warm for 55-65 minutes; the filtration is performed by vacuum filtration until there is no droplet at all.

10. The process for preparing nicotinamide mononucleotide according to claim 1, characterized in that: The temperature of the reduced pressure drying in step (5) is 25°C-30°C and the vacuum degree is -0.09MPa.

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