Preparation method and application of beta-nicotinamide mononucleotide

Abstract

The invention discloses a preparation method and application of beta-nicotinamide mononucleotide, and belongs to the technical field of nucleotide synthesis. Tetraacetyl ribose and ethyl nicotinate are used as raw materials, a catalyst is added for a condensation reaction, the catalyst comprises a component A and a component B, the component A is mercaptosilane modified mesoporous silica loaded trimethylsilyl trifluoromethanesulfonate, and the component B is nitrogen-doped carbon nanotube loaded stannous chloride. And after the condensation reaction, carrying out ammonolysis deacetylation reaction, phosphorylation reaction and post-treatment to obtain the beta-nicotinamide mononucleotide. According to the catalyst provided by the invention, the component A and the component B are two catalytic components loaded by different carriers, so that the catalyst is favorably filtered and recycled, the catalyst residue is reduced, double activation of a glycosyl donor and a nucleophilic reagent can be realized by adjusting the ratio of the component A to the component B and through a synergistic effect, the forward proceeding of a reaction is promoted, and the catalyst has high selectivity and is suitable for industrial production. The method is suitable for the application field of cosmetics.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of nucleotide synthesis, and particularly relates to a preparation method of beta-nicotinamide mononucleotide and application thereof. BACKGROUND

[0002] Nicotinamide mononucleotide (NMN) is a naturally occurring bioactive nucleotide, which is an inherent substance in the human body and is also rich in some fruits and vegetables. NMN has alpha and beta isomers, and the beta isomer is the active form of NMN. The intake of NMN can change the composition and activity of intestinal microorganisms, restore mitochondrial function, inhibit the activation of inflammatory pathways, and play the roles of improving the metabolism of the body, resisting oxidative stress and reducing inflammation, and is widely used in the field of medical anti-aging.

[0003] The synthesis method of NMN includes chemical synthesis method and biological synthesis method. The chemical synthesis method realizes the large-scale production of NMN at an earlier time, and mainly uses adenosine monophosphate, tetraacetyl ribose, nicotinamide (NAM) and the like as raw materials to complete through different synthesis steps. Compared with the chemical synthesis method, the biological synthesis method has the advantages of high stereoselectivity, mild reaction conditions, fewer by-products, higher product purity and the like. Because it does not contain organic solvent residues and chirality problems, and the prepared NMN has the same configuration as the body, it becomes the current green and environmentally friendly preparation method of NMN.

[0004] However, the biological synthesis method has high cost, and the source of raw materials is limited, which cannot realize large-scale industrial production. Optimizing and improving the chemical synthesis method to overcome the defect of low purity is an important research topic for the industrial development of the synthesis of beta-nicotinamide mononucleotide. SUMMARY

[0005] The application provides a preparation method of beta-nicotinamide mononucleotide and application thereof, which can solve the problem of low purity of beta-nicotinamide mononucleotide in the prior art.

[0006] The purpose of the application can be achieved by the following technical solutions.

[0007] A preparation method of beta-nicotinamide mononucleotide, comprising the following steps:

[0008] S1, tetraacetyl ribose is dissolved in a solvent, a catalyst and nicotinyl ethyl ester are added for condensation reaction, and after the catalyst is removed by filtration, nicotinyl ethyl ester triacetyl nucleoside is obtained by reduced pressure distillation;

[0009] S2, the nicotinyl ethyl ester triacetyl nucleoside is added into methanol, and deacetylation reaction is carried out by introducing ammonia, and nicotinamide nucleoside salt is obtained by reduced pressure distillation;

[0010] S3, nicotinamide riboside salt and phosphoric acid trimethyl ester mixed solution, dropwise add phosphorus oxychloride, carry out phosphorylation reaction, after treatment after reaction end obtain beta-nicotinamide mononucleotide;

[0011] The catalyst comprises an A component and a B component, the A component is mesoporous silica modified by mercapto silane loaded with trimethylsilyl triflate, and the B component is nitrogen-doped carbon nanotube loaded with stannous chloride.

[0012] In a conventional chemical synthesis method of beta-nicotinamide mononucleotide, tetraacetyl ribose and ethyl nicotinate are used as raw materials, and beta-nicotinamide mononucleotide is prepared through processes such as condensation, deacetylation, phosphorylation and ammonolysis; in the condensation stage, trimethylsilyl triflate (TMSOTf) is often used as a catalyst, but when TMSOTf is used alone as the catalyst, the condensation reaction is prone to cause racemization of the product, and a certain proportion of alpha isomers exists in the product, and the alpha isomers are difficult to separate subsequently, so that the purity of the product is low.

[0013] The A component in the catalyst provided by the application uses the high specific surface area porous structure of mesoporous silica to load trimethylsilyl triflate, and the mercapto (-SH) modified mesoporous silica mercapto (-SH) plays an anchoring role for TMSOTf, which is helpful to the dispersion of the catalyst in the reaction system; the mesoporous structure provides a large specific surface area and ordered pores, which not only can load TMSOTf, but also is conducive to the diffusion of reactants and the exposure of active sites. Unlike the traditional direct addition of trimethylsilyl triflate, the A component has better stability and is convenient for recycling.

[0014] The B component in the catalyst has stable structure, and the loading limits the excessive reaction of the active component, thereby reducing byproducts. The A component and the B component have a synergistic effect; the trimethylsilyl triflate of the A component is coordinated and activated with the carbonyl oxygen at the C1 position of tetraacetyl ribose, so that the C1 position forms a highly electron-deficient carbon cation intermediate, the generated C1 position carbon cation intermediate is more easily attacked by the nitrogen atom (nucleophilic site) in the pyridine ring of ethyl nicotinate, and based on the steric effect, the beta isomer is forced to be generated with high selectivity, and the pi-pi effect of the nitrogen-doped carbon nanotube is also conducive to the adsorption of ethyl nicotinate; the stannous ion in stannous chloride is in a relatively low oxidation state and has electron-donating ability; the ester group directly connected to the pyridine ring is a strong electron-withdrawing group; a weak coordination bond is formed by induction and gradually approaches; the stannous ion transfers electrons to the ester group, so that the electron cloud density of the nitrogen atom in the pyridine ring is increased, the nucleophilicity of the nitrogen atom is enhanced, the nucleophilic attack of the pyridine nitrogen on the C1 position carbon cation intermediate is promoted, and the product is stabilized. The “double activation” of the glycosyl donor and the nucleophile is realized, the reaction efficiency and the yield are greatly improved, the forward progress of the reaction is promoted, and the purity of the product is improved. In addition, after the coordination of SnCl2 and ethyl nicotinate in the reaction, the rigid structure of the loading carrier and Sn 2+The coordination conformation of the pyridine ring further limits the access angle of ethyl nicotinate, effectively promoting high β isomer selectivity.

[0015] Further, the solvent is one of dichloromethane, chloroform, trichloromethane and acetonitrile.

[0016] Further, the mass-volume ratio of the tetraacetyl ribose and the solvent is 100-150 g / L.

[0017] Further, the mass ratio of the A component and the B component in the catalyst is 1: (0.5-2);

[0018] The addition amount of the catalyst is 1.5-2 times the mass of ethyl nicotinate;

[0019] The molar ratio of ethyl nicotinate and tetraacetyl ribose is 1-3:1.

[0020] Further, the preparation steps of the A component in the catalyst are as follows:

[0021] Step one, disperse mesoporous silica in ethanol to form a dispersion with a concentration of 10-20 mg / mL;

[0022] Step two, add the dispersion to an ethanol aqueous solution with an ethanol volume fraction of 30-70%, obtain a mixed solution, add 3-mercaptopropyltrimethoxysilane, add ammonia water to adjust the pH to 11.5-12.5, react for 12-18 h, centrifuge, wash and dry to obtain mercaptosilane-modified mesoporous silica;

[0023] Step three, disperse the mercaptosilane-modified mesoporous silica in anhydrous dichloromethane under an inert atmosphere, add trimethylsilyl triflate dropwise under ice bath, remove the ice bath after the addition is completed, warm up to 30-40℃ and stir for 4-8 h, centrifuge, wash and dry to obtain the A component.

[0024] Mesoporous silica is a kind of nanomaterial with high specific surface area and ordered pore structure, 3-mercaptopropyltrimethoxysilane is hydrolyzed and condensed in alcohol aqueous solution to achieve the purpose of modifying silica, mesoporous silica is rich in mercapto groups, which enhances the loading firmness of trimethylsilyl triflate. Trimethylsilyl triflate has strong volatility, so the reaction conditions need to be strictly controlled during loading.

[0025] Further, the particle size of the mesoporous silica is 50-200 nm.

[0026] Further, the concentration of mesoporous silica in the mixed solution is 3-6 mg / mL.

[0027] Further, the 3-mercaptopropyltrimethoxysilane is added at a concentration of 15-25 mM.

[0028] Further, the mass-volume ratio of the mercapto-silane modified mesoporous silica and anhydrous dichloromethane is 10-20 mg / mL.

[0029] The mass ratio of the trimethylsilyl trifluoromethanesulfonate and the mercapto-silane modified mesoporous silica is (0.1-0.5):1.

[0030] Further, the preparation steps of the B component in the catalyst are as follows:

[0031] Step 1, dispersing the nitrogen-doped carbon nanotubes in anhydrous ethanol to form a suspension;

[0032] Step 2, adding anhydrous stannous chloride to the suspension under an inert atmosphere, stirring at 80-90℃ for 6-12h, centrifuging and washing, and then drying to obtain the B component.

[0033] The anhydrous stannous chloride is easy to hydrolyze and oxidize, and the anhydrous and oxygen-free conditions should be strictly controlled during the reaction.

[0034] Further, the concentration of the nitrogen-doped carbon nanotubes in the suspension is 10-30 mg / mL.

[0035] The mass ratio of the nitrogen-doped carbon nanotubes and the anhydrous stannous chloride is 1:(3-6).

[0036] Further, the mass-volume ratio of the ethyl nicotinate triacetyl nucleoside and methanol is 0.1-0.2 kg / L.

[0037] The molar ratio of the ammonia gas and the ethyl nicotinate triacetyl nucleoside is (2-4):1.

[0038] Further, the temperature of the deacetylation reaction is-5℃-0℃, and the reaction time is 12-24h.

[0039] Further, the mass-volume ratio of the nicotinamide nucleoside salt and trimethyl phosphate is 0.1-0.2 kg / L.

[0040] The molar ratio of the nicotinamide nucleoside salt and phosphorus oxychloride is 1:(1.5-2).

[0041] Further, the temperature of the phosphorylation reaction is-10℃--5℃, and the reaction time is 8-12h.

[0042] The application also provides the application of the β-nicotinamide mononucleotide prepared by the preparation method of the β-nicotinamide mononucleotide in cosmetic materials.

[0043] The application has the following beneficial effects:

[0044] (1) The present application uses tetraacetyl ribose and ethyl nicotinate as raw materials to prepare beta-nicotinamide mononucleotide, provides a supported catalyst to participate in the condensation reaction, utilizes the synergistic effect of component A (mercapto silane modified mesoporous silica supported trimethylsilylethyl trifluoromethanesulfonate) and component B (nitrogen-doped carbon nanotube supported stannous chloride) to realize the "double activation" of the glycosyl donor and the nucleophile, greatly improve the reaction efficiency and yield, promote the forward progress of the reaction, and improve the high selectivity of beta isomer and the purity of the product.

[0045] (2) The catalyst of the present application is supported on different carriers (mesoporous silica and nitrogen-doped carbon nanotubes), which on the one hand makes the active components (trimethylsilylethyl trifluoromethanesulfonate and stannous chloride) more stable, avoids side reactions caused by excessive action of free active components, and on the other hand facilitates the filtration and recycling of the catalyst after being supported, reduces catalyst residues, simplifies the subsequent purification steps, and improves the purity of the product.

[0046] (3) The present application can control the activity of the reactants by adjusting the ratio of component A and component B in the catalyst, reduce by-products, and improve the selectivity and efficiency of the reaction. DETAILED DESCRIPTION

[0047] The technical solutions in the embodiments of the present application will be described below in conjunction with the embodiments of the present application. Obviously, the described embodiments are only a part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of the present application.

[0048] Example 1

[0049] Preparation of component A in the catalyst:

[0050] Step one, disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion liquid with a concentration of 10 mg / mL.

[0051] Step two, add the dispersion liquid to the ethanol aqueous solution with a volume fraction of 50% ethanol to obtain a mixed liquid, the concentration of mesoporous silica in the mixed liquid is 5 mg / mL, 3-mercaptopropyltrimethoxysilane is added according to a concentration of 15 mM, ammonia water is added to adjust the pH to 12, the reaction is carried out for 18 h, and then the product is washed with ethanol and deionized water in sequence and vacuum dried to obtain mercaptosilane modified mesoporous silica.

[0052] Step three, the mercapto silane modified mesoporous silica is dispersed in anhydrous dichloromethane under nitrogen atmosphere at a concentration of 10 mg / mL, trimethylsilyl triflate is added dropwise under ice bath, the mass ratio of trimethylsilyl triflate to mercapto silane modified mesoporous silica is 0.1:1, after the dropwise addition is completed, the ice bath is removed, the temperature is increased to 40℃, and the reaction is stirred for 6 hours, the precipitate is obtained by centrifugation, washed with anhydrous ethanol, and dried to obtain component A.

[0053] Preparation of component B in the catalyst:

[0054] Step 1, the nitrogen-doped carbon nanotubes are dispersed in anhydrous ethanol to form a suspension with a concentration of 20 mg / mL.

[0055] Step 2, under nitrogen atmosphere, anhydrous stannous chloride is added to the suspension, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is 1:3, the reaction is stirred at 80℃ for 12 hours, and after centrifugation and washing, component B is obtained by drying.

[0056] Preparation of β-nicotinamide mononucleotide:

[0057] S1, tetraacetyl ribose is dissolved in dichloromethane at a concentration of 100 g / L, catalyst (the mass ratio of component A to component B is 1:0.5) and ethyl nicotinate are added, the amount of catalyst added is 1.5 times the mass of ethyl nicotinate, the molar ratio of ethyl nicotinate to tetraacetyl ribose is 1:1, the condensation reaction is carried out under constant temperature of 30℃, after 2 hours of reaction, the catalyst is removed by filtration, and the solvent is removed from the filtrate using a rotary evaporator under vacuum of-0.09 MPa to obtain ethyl nicotinate triacetyl nucleoside.

[0058] S2, the obtained ethyl nicotinate triacetyl nucleoside is added to methanol at a concentration of 0.1 kg / L, after stirring and dissolving, the temperature is lowered to-5℃, ammonia gas is introduced, the molar ratio of ammonia gas to ethyl nicotinate triacetyl nucleoside is 3:1, the deacetylation reaction is carried out by increasing the temperature to 0℃, the reaction is carried out for 12 hours, and the solvent is removed using a rotary evaporator under vacuum of-0.09 MPa to obtain nicotinamide nucleoside salt.

[0059] S3, nicotinamide nucleoside salt and trimethyl phosphate are mixed and dissolved, the concentration of nicotinamide nucleoside salt is 0.1 kg / L, the temperature is lowered to-10℃, and phosphorus oxychloride is added dropwise, the molar ratio of nicotinamide nucleoside salt to phosphorus oxychloride is 1:1.5, the phosphorylation reaction is carried out by stirring at constant temperature of-10℃, the reaction is carried out for 10 hours, after the reaction is completed, distilled water is added and stirred, ethyl acetate is further added and stirred, after the liquid is separated into oil phase and water phase, the oil phase is removed, the water phase is passed through a filter filled with D202 resin to remove ammonium salt, and vacuum drying is carried out to obtain β-nicotinamide mononucleotide.

[0060] The β-nicotinamide mononucleotide sample is collected for purity test, the purity is 98.1%, and the yield is 81.6%.

[0061] Example 2

[0062] The difference from Example 1 is that in Step two of preparing component A of the catalyst, 3-mercaptopropyltrimethoxysilane is added at a concentration of 15 mM, which is adjusted to be added at a concentration of 20 mM.

[0063] The steps for preparing component A of the catalyst are as follows, and other conditions and steps are the same as in Example 1.

[0064] Step one, disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion with a concentration of 10 mg / mL.

[0065] Step two, add the dispersion to an ethanol aqueous solution with a volume fraction of 50% ethanol to obtain a mixed solution, the concentration of mesoporous silica in the mixed solution is 5 mg / mL, 3-mercaptopropyltrimethoxysilane is added at a concentration of 20 mM, ammonia is added to adjust the pH to 12, and the reaction is carried out for 18 h. After centrifugation, wash with ethanol and deionized water in turn, and vacuum dry to obtain mercaptosilane-modified mesoporous silica.

[0066] Step three, disperse the mercaptosilane-modified mesoporous silica in anhydrous dichloromethane under a nitrogen atmosphere at a concentration of 10 mg / mL, and add trimethylsilyl triflate dropwise in an ice bath. The mass ratio of trimethylsilyl triflate to mercaptosilane-modified mesoporous silica is 0.1:1. After the addition is complete, remove the ice bath and warm to 40°C for stirring for 6 h. Centrifuge to obtain the precipitate, wash with anhydrous ethanol, and dry to obtain component A.

[0067] Collect the β-nicotinamide mononucleotide sample to test the purity, the purity is 98.5%, and the yield is 82.0%.

[0068] Example 3

[0069] The difference from Example 1 is that in Step two of preparing component A of the catalyst, 3-mercaptopropyltrimethoxysilane is added at a concentration of 15 mM, which is adjusted to be added at a concentration of 25 mM.

[0070] The steps for preparing component A of the catalyst are as follows, and other conditions and steps are the same as in Example 1.

[0071] Step one, disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion with a concentration of 10 mg / mL.

[0072] Step 2: Add the dispersion to an ethanol aqueous solution with a volume ratio of 50% to obtain a mixture. The concentration of mesoporous silica in the mixture is 5 mg / mL. Add 3-mercaptopropyltrimethoxysilane at a concentration of 25 mM and mix. Add ammonia to adjust the pH to 12. React for 18 h. After centrifugation, wash with ethanol and deionized water in sequence, and vacuum dry to obtain mercaptosilane-modified mesoporous silica.

[0073] Step 3: Disperse the mercaptosilane-modified mesoporous silica at a concentration of 10 mg / mL in anhydrous dichloromethane under a nitrogen atmosphere. Add trimethylsilyl trifluoromethanesulfonate dropwise under an ice bath. The mass ratio of trimethylsilyl trifluoromethanesulfonate to mercaptosilane-modified mesoporous silica is 0.1:1. After the addition is complete, remove the ice bath, heat to 40°C and stir for 6 hours. Centrifuge, take the precipitate, wash with anhydrous ethanol and dry to obtain component A.

[0074] The purity of β-nicotinamide mononucleotide samples was tested and found to be 98.4%, with a yield of 82.0%.

[0075] Example 4

[0076] The only difference from Example 2 is that in step three of preparing catalyst component A, the mass ratio of trimethylsilyl trifluoromethanesulfonate and mercaptosilane-modified mesoporous silica was adjusted from 0.1:1 to 0.3:1.

[0077] The steps for preparing component A in the catalyst are as follows, with other conditions and steps the same as in Example 1.

[0078] Step 1: Disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion with a concentration of 10 mg / mL.

[0079] Step 2: Add the dispersion to an ethanol aqueous solution with a volume ratio of 50% to obtain a mixture. The concentration of mesoporous silica in the mixture is 5 mg / mL. Add 3-mercaptopropyltrimethoxysilane at a concentration of 20 mM and mix. Add ammonia to adjust the pH to 12. React for 18 h. After centrifugation, wash with ethanol and deionized water in sequence, and vacuum dry to obtain mercaptosilane-modified mesoporous silica.

[0080] Step 3: Disperse the mercaptosilane-modified mesoporous silica at a concentration of 10 mg / mL in anhydrous dichloromethane under a nitrogen atmosphere. Add trimethylsilyl trifluoromethanesulfonate dropwise under an ice bath. The mass ratio of trimethylsilyl trifluoromethanesulfonate to mercaptosilane-modified mesoporous silica is 0.3:1. After the addition is complete, remove the ice bath, heat to 40°C and stir for 6 hours. Centrifuge, take the precipitate, wash with anhydrous ethanol and dry to obtain component A.

[0081] The purity of β-nicotinamide mononucleotide samples was tested and found to be 98.8%, with a yield of 82.4%.

[0082] Example 5

[0083] The difference from Example 2 is that in Step 3 of preparing Component A of the catalyst, the mass ratio of trimethylsilyl trifluoromethanesulfonate to the mercapto-silane modified mesoporous silica is adjusted from 0.1:1 to 0.5:1.

[0084] The steps for preparing Component A of the catalyst are as follows, and other conditions and steps are the same as in Example 1.

[0085] Step 1, disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion with a concentration of 10 mg / mL.

[0086] Step 2, add the dispersion to an ethanol aqueous solution with a volume fraction of 50% ethanol to obtain a mixture, the concentration of mesoporous silica in the mixture is 5 mg / mL, add 3-mercaptopropyltrimethoxysilane according to a concentration of 20 mM, add ammonia water to adjust the pH to 12, react for 18 h, centrifuge, then wash with ethanol and deionized water in sequence, and vacuum dry to obtain mercapto-silane modified mesoporous silica.

[0087] Step 3, disperse the mercapto-silane modified mesoporous silica in anhydrous dichloromethane under a nitrogen atmosphere at a concentration of 10 mg / mL, add trimethylsilyl trifluoromethanesulfonate dropwise in an ice bath, the mass ratio of trimethylsilyl trifluoromethanesulfonate to the mercapto-silane modified mesoporous silica is 0.5:1, remove the ice bath after the dropwise addition is completed, warm up to 40°C, and stir for 6 h, centrifuge to obtain the precipitate, wash with anhydrous ethanol, and dry to obtain Component A.

[0088] Collect the β-nicotinamide mononucleotide sample to test the purity, the purity is 98.7%, and the yield is 82.2%.

[0089] Example 6

[0090] The difference from Example 4 is that in Step 2 of preparing Component B of the catalyst, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is adjusted from 1:3 to 1:5.

[0091] The steps for preparing Component B of the catalyst are as follows, and other conditions and steps are the same as in Example 4.

[0092] Step 1, disperse nitrogen-doped carbon nanotubes in anhydrous ethanol to form a suspension with a concentration of 20 mg / mL.

[0093] Step 2, add anhydrous stannous chloride to the suspension under a nitrogen atmosphere, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is 1:5, stir at 80°C for 12 h, centrifuge and wash, and then dry to obtain Component B.

[0094] The purity of the collected β-nicotinamide mononucleotide sample was tested, and the purity was 98.9%, and the yield was 82.6%.

[0095] Example 7

[0096] The difference from Example 4 is only that in Step 2 of preparing Component B of the catalyst, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is adjusted from 1:3 to 1:6.

[0097] The steps for preparing Component B of the catalyst are as follows, and the other conditions and steps are the same as in Example 4.

[0098] Step 1, disperse nitrogen-doped carbon nanotubes in anhydrous ethanol to form a suspension with a concentration of 20 mg / mL.

[0099] Step 2, add anhydrous stannous chloride to the suspension under a nitrogen atmosphere, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is 1:6, stir at 80°C for 12h, centrifugal wash and dry to obtain Component B.

[0100] The purity of the collected β-nicotinamide mononucleotide sample was tested, and the purity was 98.9%, and the yield was 82.6%.

[0101] Example 8

[0102] The difference from Example 6 is only that the mass ratio of Component A and Component B of the catalyst for preparing β-nicotinamide mononucleotide is adjusted to 1:1.5.

[0103] Component A of the catalyst:

[0104] Step 1, disperse mesoporous silica with a particle size of 50-200 nm in ethanol to form a dispersion with a concentration of 10 mg / mL.

[0105] Step 2, add the dispersion to an ethanol aqueous solution with a volume fraction of 50% ethanol to obtain a mixture, the concentration of mesoporous silica in the mixture is 5 mg / mL, and 3-mercaptopropyl trimethoxysilane is added according to a concentration of 20 mM, ammonia water is added to adjust the pH to 12, and the reaction is carried out for 18h, then centrifugal wash with ethanol and deionized water in turn, and vacuum drying to obtain mercaptosilane-modified mesoporous silica.

[0106] Step 3, disperse the mercaptosilane-modified mesoporous silica in anhydrous dichloromethane under a nitrogen atmosphere at a concentration of 10 mg / mL, and drop 3-trimethylsilyl trifluoromethanesulfonate in an ice bath, the mass ratio of 3-trimethylsilyl trifluoromethanesulfonate to mercaptosilane-modified mesoporous silica is 0.3:1, after the drop is completed, remove the ice bath, and warm up to 40°C for stirring reaction for 6h, centrifugal take the precipitate, wash with anhydrous ethanol, and dry to obtain Component A.

[0107] Component B of the catalyst:

[0108] Step 1, nitrogen-doped carbon nanotubes were dispersed in anhydrous ethanol to form a suspension with a concentration of 20 mg / mL.

[0109] Step 2, under a nitrogen atmosphere, anhydrous stannous chloride was added to the suspension, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride was 1:5, and the reaction was stirred at 80°C for 12 h. After centrifugal washing and drying, component B was obtained.

[0110] Preparation of β-nicotinamide mononucleotide:

[0111] S1, tetraacetyl ribose was dissolved in dichloromethane with a concentration of 100 g / L, a catalyst (the mass ratio of component A to component B was 1:1.5) and ethyl nicotinate were added, the amount of catalyst added was 1.5 times the mass of ethyl nicotinate, the molar ratio of ethyl nicotinate to tetraacetyl ribose was 1:1, and the condensation reaction was carried out under constant temperature of 30°C with stirring, after 2 h of reaction, the catalyst was removed by filtration, and the filtrate was subjected to solvent removal using a rotary evaporator under vacuum of-0.09 MPa to obtain ethyl nicotinate triacetyl nucleoside.

[0112] S2, the obtained ethyl nicotinate triacetyl nucleoside was added to methanol with a concentration of 0.1 kg / L, after stirring and dissolving, the temperature was lowered to-5°C, ammonia gas was introduced, the molar ratio of ammonia gas to ethyl nicotinate triacetyl nucleoside was 3:1, the temperature was raised to 0°C for deacetylation reaction, the reaction was carried out for 12 h, and the solvent was removed using a rotary evaporator under vacuum of-0.09 MPa to obtain nicotinamide nucleoside salt.

[0113] S3, nicotinamide nucleoside salt and trimethyl phosphate were mixed and dissolved, the concentration of nicotinamide nucleoside salt was 0.1 kg / L, the temperature was lowered to-10°C, phosphorus oxychloride was added dropwise, the molar ratio of nicotinamide nucleoside salt to phosphorus oxychloride was 1:1.5, the phosphorylation reaction was carried out under constant temperature of-10°C with stirring, the reaction was carried out for 10 h, after the reaction was completed, distilled water was added and stirred, then ethyl acetate was added and stirred, after standing and separating into oil and water phases, the oil phase was removed, the water phase was passed through a filter filled with D202 resin to remove ammonium salt, and vacuum drying was carried out to obtain β-nicotinamide mononucleotide.

[0114] The β-nicotinamide mononucleotide sample was collected for purity test, the purity was 99.2%, and the yield was 83.0%.

[0115] Example 9

[0116] The difference from Example 6 is that the mass ratio of component A to component B in the catalyst for preparing β-nicotinamide mononucleotide is adjusted to 1:2.

[0117] Preparation of component A in the catalyst:

[0118] Step one, disperse the mesoporous silica with particle size of 50-200nm in ethanol to form a dispersion with concentration of 10mg / mL.

[0119] Step two, add the dispersion to the ethanol aqueous solution with volume ratio of ethanol of 50% to obtain a mixed solution, the concentration of mesoporous silica in the mixed solution is 5mg / mL, add 3-mercaptopropyl trimethoxysilane according to the concentration of 20mM, add ammonia water to adjust the pH to 12, react for 18h, centrifuge, then wash with ethanol and deionized water in turn, and vacuum dry to obtain mercapto silane modified mesoporous silica.

[0120] Step three, disperse the mercapto silane modified mesoporous silica in anhydrous dichloromethane under nitrogen atmosphere at a concentration of 10mg / mL, add trimethylsilyl triflate dropwise under ice bath, the mass ratio of trimethylsilyl triflate to mercapto silane modified mesoporous silica is 0.3:1, remove the ice bath after the dropwise addition is completed, warm up to 40℃ and stir for 6h, centrifuge to obtain the precipitate, wash with anhydrous ethanol and dry to obtain component A.

[0121] Preparation of component B in the preparation of the catalyst:

[0122] Step 1, disperse the nitrogen-doped carbon nanotubes in anhydrous ethanol to form a suspension with a concentration of 20mg / mL.

[0123] Step 2, add anhydrous stannous chloride to the suspension under nitrogen atmosphere, the mass ratio of nitrogen-doped carbon nanotubes to anhydrous stannous chloride is 1:5, stir at 80℃ for 12h, centrifuge and dry after washing to obtain component B.

[0124] Preparation of β-nicotinamide mononucleotide:

[0125] S1, dissolve tetraacetyl ribose in dichloromethane with a concentration of 100g / L, add catalyst (the mass ratio of component A to component B is 1:2) and ethyl nicotinate, the amount of catalyst added is 1.5 times the mass of ethyl nicotinate, the molar ratio of ethyl nicotinate to tetraacetyl ribose is 1:1, stir for condensation reaction at a constant temperature of 30℃, after 2h of reaction, remove the catalyst by filtration, and remove the solvent from the filtrate using a rotary evaporator at a vacuum degree of-0.09MPa to obtain ethyl nicotinate triacetyl nucleoside.

[0126] S2, add the obtained ethyl nicotinate triacetyl nucleoside to methanol with a concentration of 0.1kg / L, stir and dissolve, then cool to-5℃, pass in ammonia gas, the molar ratio of ammonia gas to ethyl nicotinate triacetyl nucleoside is 3:1, warm up to 0℃ for deacetylation reaction, react for 12h, remove the solvent using a rotary evaporator at a vacuum degree of-0.09MPa to obtain nicotinamide nucleoside salt.

[0127] S3, nicotinamide riboside salt and phosphoric acid trimethyl ester are mixed and dissolved, the concentration of nicotinamide riboside salt is 0.1 kg / L, the temperature is reduced to -10℃, and phosphorus oxychloride is added dropwise, the molar ratio of nicotinamide riboside salt to phosphorus oxychloride is 1:1.5, the temperature is kept at -10℃, and the phosphorylation reaction is carried out under stirring, the reaction is carried out for 10 hours, after the reaction is completed, distilled water is added and stirred, then ethyl acetate is added and stirred, after standing and layering, the oil phase layer is removed, the water phase layer is removed through a filter filled with D202 resin to remove ammonium salt, and vacuum drying is performed to obtain β-nicotinamide mononucleotide.

[0128] The purity of the collected β-nicotinamide mononucleotide sample is tested, the purity is 99.1%, and the yield is 82.8%.

[0129] Comparative Example 1

[0130] The difference from Example 1 is that only trimethylsilyl trifluoromethanesulfonate is used as a catalyst.

[0131] Preparation of β-nicotinamide mononucleotide:

[0132] S1, tetraacetyl ribose is dissolved in dichloromethane, the concentration is 100 g / L, nicotinic acid ethyl ester is added, trimethylsilyl trifluoromethanesulfonate is added dropwise at room temperature, the molar ratio of nicotinic acid ethyl ester to tetraacetyl ribose is 1:1, the amount of trimethylsilyl trifluoromethanesulfonate added is 1.5 times the mass of nicotinic acid ethyl ester, the condensation reaction is carried out under stirring at a constant temperature of 30℃, after 2 hours of reaction, the solvent is removed by a rotary evaporator under a vacuum degree of -0.09 MPa to obtain nicotinic acid ethyl ester triacetyl riboside.

[0133] S2, the obtained nicotinic acid ethyl ester triacetyl riboside is added to methanol, the concentration is 0.1 kg / L, after stirring and dissolving, the temperature is reduced to -5℃, ammonia gas is introduced, the molar ratio of ammonia gas to nicotinic acid ethyl ester triacetyl riboside is 3:1, the deacetylation reaction is carried out by increasing the temperature to 0℃, the reaction is carried out for 12 hours, the solvent is removed by a rotary evaporator under a vacuum degree of -0.09 MPa to obtain nicotinamide riboside salt.

[0134] S3, nicotinamide riboside salt and phosphoric acid trimethyl ester are mixed and dissolved, the concentration of nicotinamide riboside salt is 0.1 kg / L, the temperature is reduced to -10℃, and phosphorus oxychloride is added dropwise, the molar ratio of nicotinamide riboside salt to phosphorus oxychloride is 1:1.5, the temperature is kept at -10℃, and the phosphorylation reaction is carried out under stirring, the reaction is carried out for 10 hours, after the reaction is completed, distilled water is added and stirred, then ethyl acetate is added and stirred, after standing and layering, the oil phase layer is removed, the water phase layer is removed through a filter filled with D202 resin to remove ammonium salt, and vacuum drying is performed to obtain β-nicotinamide mononucleotide.

[0135] The purity of the collected β-nicotinamide mononucleotide sample is tested, the purity is 96.3%, and the yield is 71.0%.

[0136] Comparative Example 2

[0137] The difference from Example 1 is that this comparative example directly uses trimethylsilyl trifluoromethanesulfonate and stannous chloride as catalysts.

[0138] Preparation of β-nicotinamide mononucleotide:

[0139] S1, tetraacetyl ribose was dissolved in dichloromethane with a concentration of 100 g / L, and ethyl nicotinate was added. Stannous chloride anhydrous was dissolved in anhydrous ethanol with a concentration of 0.1 g / mL. Trimethylsilyl trifluoromethanesulfonate and anhydrous stannous chloride ethanol solution were added dropwise at room temperature. The molar ratio of ethyl nicotinate to tetraacetyl ribose was 1:1. The total mass of trimethylsilyl trifluoromethanesulfonate and anhydrous stannous chloride was 1.5 times the mass of ethyl nicotinate. The mass ratio of trimethylsilyl trifluoromethanesulfonate to anhydrous stannous chloride was 1:0.5. The condensation reaction was carried out at a constant temperature of 30°C with stirring. After 2 hours of reaction, the solvent was removed using a rotary evaporator at a vacuum degree of -0.09 MPa to obtain ethyl nicotinate triacetyl nucleoside.

[0140] S2, the obtained ethyl nicotinate triacetyl nucleoside was added to methanol with a concentration of 0.1 kg / L. After stirring and dissolving, the temperature was lowered to -5°C, and ammonia gas was introduced. The molar ratio of ammonia gas to ethyl nicotinate triacetyl nucleoside was 3:1. The deacetylation reaction was carried out by warming to 0°C. After 12 hours of reaction, the solvent was removed using a rotary evaporator at a vacuum degree of -0.09 MPa to obtain nicotinamide nucleoside salt.

[0141] S3, nicotinamide nucleoside salt and trimethyl phosphate were mixed and dissolved. The concentration of nicotinamide nucleoside salt was 0.1 kg / L. The temperature was lowered to -10°C, and phosphorus oxychloride was added dropwise. The molar ratio of nicotinamide nucleoside salt to phosphorus oxychloride was 1:1.5. The phosphorylation reaction was carried out at a constant temperature of -10°C with stirring. After 10 hours of reaction, distilled water was added and stirred, and then ethyl acetate was added and stirred. After the oil phase layer was removed by standing and layering, the water phase layer was removed through a filter filled with D202 resin to remove ammonium salt. Vacuum drying obtained β-nicotinamide mononucleotide.

[0142] The β-nicotinamide mononucleotide sample was collected for purity testing, with a purity of 96.6% and a yield of 75.3%.

[0143] Comparative Example 3

[0144] The difference from Example 1 is that this comparative example directly uses the A component in Example 1 as the catalyst.

[0145] Preparation of catalyst:

[0146] Step one, mesoporous silica with a particle size of 50-200 nm was dispersed in ethanol to form a dispersion liquid with a concentration of 10 mg / mL.

[0147] Step two, add the dispersion liquid to the ethanol aqueous solution with a volume ratio of 50% ethanol, to obtain a mixed solution, the concentration of mesoporous silicon dioxide in the mixed solution is 5 mg / mL, 3-mercaptopropyl trimethoxysilane is added according to a concentration of 15 mM, ammonia water is added to adjust the pH to 12, and the reaction is carried out for 18 h, then centrifugation is performed, and then the product is washed with ethanol and deionized water in sequence, and vacuum drying is performed to obtain mercapto silane modified mesoporous silicon dioxide.

[0148] Step three, the mercapto silane modified mesoporous silicon dioxide is dispersed in anhydrous dichloromethane under a nitrogen atmosphere at a concentration of 10 mg / mL, and trimethylsilyl triflate is added dropwise under ice bath, the mass ratio of trimethylsilyl triflate to mercapto silane modified mesoporous silicon dioxide is 0.1:1, after the dropwise addition is completed, the ice bath is removed, the temperature is increased to 40 DEG C, and the reaction is carried out for 6 h under stirring, then the precipitate is obtained by centrifugation, and then the precipitate is washed with anhydrous ethanol and dried to obtain the catalyst.

[0149] Preparation of beta-nicotinamide mononucleotide:

[0150] S1, tetraacetyl ribose is dissolved in dichloromethane, the concentration is 100 g / L, a catalyst and ethyl nicotinate are added, the amount of the catalyst added is 1.5 times the mass of the ethyl nicotinate, the molar ratio of the ethyl nicotinate to the tetraacetyl ribose is 1:1, the condensation reaction is carried out under stirring at a constant temperature of 30 DEG C, after the reaction is carried out for 2 h, the catalyst is removed by filtration, and then the solvent is removed from the filtrate by using a rotary evaporator under a vacuum degree of-0.09 MPa to obtain ethyl nicotinate triacetyl nucleoside.

[0151] S2, the obtained ethyl nicotinate triacetyl nucleoside is added to methanol, the concentration is 0.1 kg / L, after the stirring and dissolution, the temperature is decreased to-5 DEG C, ammonia gas is introduced, the molar ratio of the ammonia gas to the ethyl nicotinate triacetyl nucleoside is 3:1, the deacetylation reaction is carried out at a temperature of 0 DEG C, the reaction is carried out for 12 h, and then the solvent is removed by using a rotary evaporator under a vacuum degree of-0.09 MPa to obtain nicotinamide nucleoside salt.

[0152] S3, the nicotinamide nucleoside salt and trimethyl phosphate are mixed and dissolved, the concentration of the nicotinamide nucleoside salt is 0.1 kg / L, the temperature is decreased to-10 DEG C, phosphorus oxychloride is added dropwise, the molar ratio of the nicotinamide nucleoside salt to the phosphorus oxychloride is 1:1.5, the phosphorylation reaction is carried out under stirring at a constant temperature of-10 DEG C, the reaction is carried out for 10 h, after the reaction is completed, distilled water is added and stirred, then ethyl acetate is added and stirred, the oil phase layer is removed after the separation of the layers, the water phase layer is removed through a filter filled with D202 resin to remove the ammonium salt, and then vacuum drying is performed to obtain beta-nicotinamide mononucleotide.

[0153] The beta-nicotinamide mononucleotide sample is collected to test the purity, the purity is 97.1%, and the yield is 77.6%.

[0154] The above results show that the purity of the prepared beta-nicotinamide mononucleotide can reach more than 98%, and the yield can reach up to 83%, indicating that the catalyst provided by the application has better catalytic effect, the product purity is higher than that of the comparative example, and the by-products are less.

[0155] It should be noted that, in this document, relational terms such as first and second and the like can be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Moreover, the terms "comprises", "comprising", or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus.

[0156] Although embodiments of the present application have been shown and described, it is to be understood that various modifications, substitutions, alternatives, and variations can be made in the embodiments without departing from the spirit and scope of the present application as defined by the appended claims and their equivalents.

Claims

1. A method for preparing β-nicotinamide mononucleotide, characterized in that, Includes the following steps: S1, tetraacetylribose is dissolved in a solvent, a catalyst is added and ethyl nicotinic acid is added to carry out a condensation reaction, and the mixture is filtered and distilled under reduced pressure to obtain ethyl nicotinic acid triacetyl nucleoside. S2. Nicotinic acid ethyl ester triacetyl nucleoside was added to methanol, and ammonia gas was introduced to carry out a deacetylation reaction. The result was obtained by vacuum distillation to obtain nicotinamide nucleoside salt. S3, nicotinamide nucleoside salt and trimethyl phosphate were mixed and dissolved, and phosphoric acid oxychloride was added dropwise to carry out the phosphorylation reaction. After the reaction was completed, post-treatment was performed to obtain β-nicotinamide mononucleotide. The catalyst comprises component A and component B. Component A is trimethylsilyl trifluoromethanesulfonate supported on mesoporous silica modified with mercaptosilane, and component B is stannous chloride supported on nitrogen-doped carbon nanotubes.

2. The method for preparing β-nicotinamide mononucleotide according to claim 1, characterized in that, The solvent is one of dichloromethane, chloroform, trichloromethane, and acetonitrile; The mass-to-volume ratio of the tetraacetylribose to the solvent is 100–150 g / L.

3. The method for preparing β-nicotinamide mononucleotide according to claim 1, characterized in that, The mass ratio of component A to component B in the catalyst is 1:(0.5-2); The amount of catalyst added is 1.5 to 2 times the mass of ethyl nicotinic acid; The molar ratio of ethyl nicotinic acid to tetraacetyl ribose is (1-3):

1.

4. The method for preparing β-nicotinamide mononucleotide according to claim 1, characterized in that, The preparation steps of component A in the catalyst are as follows: Step 1: Disperse mesoporous silica in ethanol to form a dispersion with a concentration of 10-20 mg / mL; Step 2: Add the dispersion to an ethanol aqueous solution with a volume ratio of 30-70% to obtain a mixture. Add 3-mercaptopropyltrimethoxysilane and mix. Add ammonia to adjust the pH to 11.5-12.

5. React for 24 hours. After centrifugation, wash and dry to obtain mercaptosilane-modified mesoporous silica. Step 3: Disperse the mercaptosilane-modified mesoporous silica in anhydrous dichloromethane under an inert atmosphere, add trimethylsilyl trifluoromethanesulfonate dropwise under an ice bath, remove the ice bath after the addition is complete, heat to 30-40℃ and stir for 4-8 hours, centrifuge, wash and dry to obtain component A.

5. The method for preparing β-nicotinamide mononucleotide according to claim 4, characterized in that, In step two, the concentration of mesoporous silica in the mixture is 3–6 mg / mL; The 3-mercaptopropyltrimethoxysilane was added at a concentration of 15–25 mM.

6. The method for preparing β-nicotinamide mononucleotide according to claim 4, characterized in that, The mass-to-volume ratio of the mercaptosilane-modified mesoporous silica to anhydrous dichloromethane is 10–20 mg / mL. The mass ratio of trimethylsilyl trifluoromethanesulfonate to mercaptosilane-modified mesoporous silica is 0.1–0.5:

1.

7. The method for preparing β-nicotinamide mononucleotide according to claim 1, characterized in that, The preparation steps of component B in the catalyst are as follows: Step 1: Disperse nitrogen-doped carbon nanotubes in anhydrous ethanol to form a suspension; Step 2: Add anhydrous stannous chloride to the suspension under an inert atmosphere, stir and react at 80-90℃ for 6-12 hours, centrifuge, wash and dry to obtain component B.

8. The method for preparing β-nicotinamide mononucleotide according to claim 7, characterized in that, The concentration of nitrogen-doped carbon nanotubes in the suspension is 10–30 mg / mL; The mass ratio of the nitrogen-doped carbon nanotubes to anhydrous stannous chloride is 1:(3-6).

9. The method for preparing β-nicotinamide mononucleotide according to claim 1, characterized in that, The mass-to-volume ratio of ethyl nicotinic acid triacetyl nucleoside to methanol is 0.1–0.2 kg / L; The molar ratio of ammonia to ethyl nicotinic acid triacetyl nucleotide is (2-4):1; The mass-to-volume ratio of the nicotinamide nucleoside salt to trimethyl phosphate is 0.1–0.2 kg / L; The molar ratio of the nicotinamide nucleoside salt to phosphorus oxychloride is 1:(1.5-2).

10. The application of β-nicotinamide mononucleotide prepared by the method of any one of claims 1-9 in cosmetic materials.