CN116751826A - Preparation process of NMN anti-aging substance - Google Patents
Preparation process of NMN anti-aging substance Download PDFInfo
- Publication number
- CN116751826A CN116751826A CN202310789714.4A CN202310789714A CN116751826A CN 116751826 A CN116751826 A CN 116751826A CN 202310789714 A CN202310789714 A CN 202310789714A CN 116751826 A CN116751826 A CN 116751826A
- Authority
- CN
- China
- Prior art keywords
- nmn
- nucleoside
- solvent
- preparing
- aging
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000126 substance Substances 0.000 title claims abstract description 29
- 230000003712 anti-aging effect Effects 0.000 title claims abstract description 26
- 238000002360 preparation method Methods 0.000 title abstract description 17
- 239000002777 nucleoside Substances 0.000 claims abstract description 60
- 150000003833 nucleoside derivatives Chemical class 0.000 claims abstract description 60
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 claims abstract description 29
- 238000000034 method Methods 0.000 claims abstract description 27
- 238000010511 deprotection reaction Methods 0.000 claims abstract description 22
- 238000005859 coupling reaction Methods 0.000 claims abstract description 19
- 239000012535 impurity Substances 0.000 claims abstract description 16
- IHNHAHWGVLXCCI-FDYHWXHSSA-N [(2r,3r,4r,5s)-3,4,5-triacetyloxyoxolan-2-yl]methyl acetate Chemical compound CC(=O)OC[C@H]1O[C@@H](OC(C)=O)[C@H](OC(C)=O)[C@@H]1OC(C)=O IHNHAHWGVLXCCI-FDYHWXHSSA-N 0.000 claims abstract description 15
- 229960003966 nicotinamide Drugs 0.000 claims abstract description 15
- 235000005152 nicotinamide Nutrition 0.000 claims abstract description 15
- 239000011570 nicotinamide Substances 0.000 claims abstract description 15
- 102000009097 Phosphorylases Human genes 0.000 claims abstract description 8
- 108010073135 Phosphorylases Proteins 0.000 claims abstract description 8
- 125000006239 protecting group Chemical group 0.000 claims abstract description 7
- 239000000047 product Substances 0.000 claims description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 238000000746 purification Methods 0.000 claims description 24
- 239000002904 solvent Substances 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 17
- 238000002425 crystallisation Methods 0.000 claims description 16
- 230000008025 crystallization Effects 0.000 claims description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical group CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 12
- 239000013078 crystal Substances 0.000 claims description 12
- 239000007787 solid Substances 0.000 claims description 11
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 9
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 239000003795 chemical substances by application Substances 0.000 claims description 9
- 238000004587 chromatography analysis Methods 0.000 claims description 9
- 238000001035 drying Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical group CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 8
- 238000011097 chromatography purification Methods 0.000 claims description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- 239000000706 filtrate Substances 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 238000007867 post-reaction treatment Methods 0.000 claims description 6
- 150000002148 esters Chemical class 0.000 claims description 4
- 150000002576 ketones Chemical class 0.000 claims description 4
- 230000002378 acidificating effect Effects 0.000 claims description 3
- 230000032683 aging Effects 0.000 claims description 3
- 230000032677 cell aging Effects 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 238000001704 evaporation Methods 0.000 claims description 3
- 230000008020 evaporation Effects 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 239000012452 mother liquor Substances 0.000 claims description 3
- 238000006386 neutralization reaction Methods 0.000 claims description 3
- 238000000638 solvent extraction Methods 0.000 claims description 3
- 238000003786 synthesis reaction Methods 0.000 abstract description 9
- 238000006366 phosphorylation reaction Methods 0.000 abstract description 6
- 230000002255 enzymatic effect Effects 0.000 abstract description 5
- 230000026731 phosphorylation Effects 0.000 abstract description 4
- DAYLJWODMCOQEW-TURQNECASA-O NMN(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(O)=O)O2)O)=C1 DAYLJWODMCOQEW-TURQNECASA-O 0.000 description 71
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 12
- 230000008569 process Effects 0.000 description 11
- 238000005265 energy consumption Methods 0.000 description 8
- 239000000463 material Substances 0.000 description 8
- OQRXBXNATIHDQO-UHFFFAOYSA-N 6-chloropyridine-3,4-diamine Chemical compound NC1=CN=C(Cl)C=C1N OQRXBXNATIHDQO-UHFFFAOYSA-N 0.000 description 4
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 4
- 102100036286 Purine nucleoside phosphorylase Human genes 0.000 description 4
- 208000012839 conversion disease Diseases 0.000 description 4
- 230000006872 improvement Effects 0.000 description 4
- 108010009099 nucleoside phosphorylase Proteins 0.000 description 4
- 230000008878 coupling Effects 0.000 description 3
- 238000010168 coupling process Methods 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000007858 starting material Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- BAWFJGJZGIEFAR-NNYOXOHSSA-O NAD(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-O 0.000 description 1
- BAWFJGJZGIEFAR-DQQFMEOOSA-N [[(2r,3s,4r,5r)-5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)O[P@@](O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-DQQFMEOOSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/30—Nucleotides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/13—Nucleic acids or derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/048—Pyridine radicals
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- General Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of NMN anti-aging substance preparation, in particular to a preparation process of NMN anti-aging substance, which comprises the following steps: coupling reaction is carried out on tetraacetylribose and nicotinamide by adopting a chemical method to generate nucleoside; deprotection treatment is carried out on the generated nucleoside, and a protecting group is removed; purifying the deprotected nucleoside to obtain purified nucleoside; reacting the purified nucleoside with a phosphorylase to produce NMN; the tetraacetylribose content is 0.01-1 mol, the nicotinamide content is 0.01-1 mol, and the phosphorylase content is 0.01-1 unit/mol nucleoside. The preparation process combines the advantages of chemical synthesis and enzymatic phosphorylation, and solves the problems of high cost of an enzymolysis method and impurity generation of the chemical synthesis method, thereby ensuring the chiral consistency of the product, reducing the production cost and improving the purity of the product.
Description
Technical Field
The invention relates to the technical field of NMN anti-aging substance preparation, in particular to a preparation process of NMN anti-aging substance.
Background
Nicotinamide mononucleotide Nicotinamide Mononucleotide (NMN) is an important bioactive substance which mainly exists in various foods and is an important precursor of NAD+ (nicotinamide dinucleotide) in human bodies, and recent researches find that NMN has wide biological activity including anti-aging, liver function improvement, brain function improvement, muscle endurance improvement and the like, so that NMN has extremely wide application prospect in the fields of foods, health-care products, medicines and the like.
However, there are two main methods of producing NMN at present, one is chemical synthesis and one is enzymatic. Chemical synthesis, while capable of producing NMN on a larger scale, often contains chiral impurities that affect the biological activity of NMN, and also increase the safety risk of the product. The enzymolysis method can obtain NMN with high purity and chiral purity, but the enzyme has high price and high production cost, and is not suitable for mass production.
Therefore, developing a new method capable of ensuring NMN quality and reducing production cost is an important task of current NMN production, and the super enzymolysis method provided by the invention is based on the background and the requirements.
Disclosure of Invention
The present invention provides a process for preparing an NMN anti-ageing substance, wherein the bond is based on the above-mentioned objects.
A process for preparing an NMN anti-ageing substance, comprising the steps of:
step one: coupling reaction is carried out on tetraacetylribose and nicotinamide by adopting a chemical method to generate nucleoside;
step two: deprotection treatment is carried out on the generated nucleoside, and a protecting group is removed;
step three: purifying the deprotected nucleoside to obtain purified nucleoside;
step four: reacting the purified nucleoside with a phosphorylase to produce NMN;
the tetraacetylribose content is 0.01-1 mol, the nicotinamide content is 0.01-1 mol, and the phosphorylase content is 0.01-1 unit/mol nucleoside.
The preparation process combines the advantages of chemical synthesis and enzymatic phosphorylation, and overcomes the problems of high cost of enzymolysis and impurity generation of chemical synthesis, thereby ensuring the chiral consistency of the product, reducing the production cost and improving the purity of the product.
Further, the coupling reaction in the first step is as follows:
a. mixing tetraacetylribose and nicotinamide in a molar ratio of 1:1 in a solvent to obtain a mixture;
b. adjusting the pH value of the mixture to be in the range of 5-9 so as to facilitate the next coupling reaction;
c. carrying out coupling reaction at 25-100 ℃ to generate nucleoside;
d. and (3) carrying out post-treatment after the reaction is finished, wherein the post-treatment comprises filtrate, washing and drying to obtain a solid nucleoside product after the coupling reaction.
Further, the deprotection treatment in the second step comprises the following steps:
a. dissolving the obtained solid nucleoside product in a solvent;
b. adding an alkaline or acidic deprotection agent, wherein the deprotection agent comprises sodium hydroxide, potassium hydroxide, hydrochloric acid or sulfuric acid;
c. deprotection reaction is carried out at the temperature of 25-100 ℃ and the pH value of 5-9, and the protective group of nucleoside is removed;
d. and (3) carrying out post-reaction treatment, wherein the post-reaction treatment comprises filtrate, neutralization, washing and drying to obtain a deprotected nucleoside product.
Further, the specific purification steps in the third step are as follows:
a. dissolving the deprotected nucleoside in a solvent;
b. solvent extraction is performed through a separating funnel to separate and remove unwanted impurities;
c. distilling the extracted solution to remove the solvent;
d. and freeze-drying the solution obtained after the solvent removal to obtain a purified nucleoside product.
The above purification steps ensure the purity of the obtained nucleoside product, providing a good quality starting material for the subsequent phosphorylation reaction.
Further, the solvent includes an alcohol, a ketone or an ester, the alcohol is methanol, the ketone is acetone, and the ester is ethyl acetate.
Further, after NMN is generated in the fourth step, NMN is subjected to secondary purification treatment, wherein the secondary purification comprises chromatographic and crystallization methods, so that the purity of NMN is improved.
Further, the chromatographic purification steps are as follows:
a. dissolving the generated NMN with ethanol;
b. passing the dissolved NMN liquid through a chromatographic column to separate NMN from other possible impurities using different affinities;
c. collecting the liquid flowing out of the chromatographic column, and detecting to identify a NMN-containing part;
d. the NMN-containing fraction is further freed from the solution by evaporation, resulting in purified NMN.
By purification through chromatography, NMN can be effectively separated from impurities which may be present, ensuring the purity and quality of the final product.
Further, the crystallization and purification steps are as follows:
a. dissolving NMN obtained after chromatographic purification with ethanol;
b. slowly reducing the temperature of the solution to enable NMN to start crystallization;
c. centrifuging or filtering, and separating NMN crystals from the mother liquor;
d. washing the NMN crystals with a suitable ethanol solvent to remove impurities that may attach to the crystal surfaces;
e. and drying the washed NMN crystals to obtain NMN products with higher purity.
Purification by crystallization can further increase the purity of the NMN product and convert it to a solid form suitable for storage and use.
Furthermore, the prepared NMN anti-aging substance is used for delaying human aging and treating diseases caused by cell aging.
The invention has the beneficial effects that:
the invention combines the advantages of chemical synthesis and enzymatic phosphorylation, i.e. the problem of high cost of an enzymolysis method is solved, the problem of consistent chirality due to the synthesis method is solved, the production cost is reduced, the highest purity and quality of the product are achieved, no impurity exists, the invention has remarkable innovation and practicability, and a novel efficient NMN anti-aging substance preparation process with low cost and high purity can be provided for the pharmaceutical industry.
The preparation process has simple operation steps, is easy to implement, is suitable for industrial production, can effectively improve the yield of NMN in actual production, meets the requirements of the market on high-quality NMN, has high purity and good quality, is suitable for preparing anti-aging medicines, health-care products, food additives and the like, and has good application prospect.
Drawings
In order to more clearly illustrate the invention or the technical solutions of the prior art, the drawings which are used in the description of the embodiments or the prior art will be briefly described, it being obvious that the drawings in the description below are only of the invention and that other drawings can be obtained from them without inventive effort for a person skilled in the art.
FIG. 1 is a schematic illustration of a process flow according to an embodiment of the present invention;
FIG. 2 is a schematic diagram of a deprotection process according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a purification step according to an embodiment of the present invention;
FIG. 4 is a schematic representation of chromatographic purification according to an embodiment of the present invention;
FIG. 5 is a schematic diagram of a crystallization purification according to an embodiment of the present invention.
Description of the embodiments
The present invention will be further described in detail with reference to specific embodiments in order to make the objects, technical solutions and advantages of the present invention more apparent.
It is to be noted that unless otherwise defined, technical or scientific terms used herein should be taken in a general sense as understood by one of ordinary skill in the art to which the present invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "upper", "lower", "left", "right", etc. are used merely to indicate relative positional relationships, which may also be changed when the absolute position of the object to be described is changed.
Examples
As shown in fig. 1-5, a process for preparing an NMN anti-aging substance comprises the steps of:
step one: coupling reaction is carried out on tetraacetylribose and nicotinamide by adopting a chemical method to generate nucleoside;
step two: deprotection treatment is carried out on the generated nucleoside, and a protecting group is removed;
step three: purifying the deprotected nucleoside to obtain purified nucleoside;
step four: reacting the purified nucleoside with a phosphorylase to produce NMN;
the tetraacetylribose content was 0.5 mole, the nicotinamide content was 0.5 mole, and the phosphorylase content was 0.5 units/mole of nucleoside.
The preparation process combines the advantages of chemical synthesis and enzymatic phosphorylation, and overcomes the problems of high cost of enzymolysis and impurity generation of chemical synthesis, thereby ensuring the chiral consistency of the product, reducing the production cost and improving the purity of the product.
The coupling reaction in step one is as follows:
a. mixing tetraacetylribose and nicotinamide in a molar ratio of 1:1 in a solvent to obtain a mixture;
b. adjusting the pH value of the mixture to be within 5 so as to facilitate the next coupling reaction;
c. carrying out coupling reaction at 25 ℃ to generate nucleoside;
d. and (3) after the reaction is finished, carrying out post-treatment, wherein the post-treatment comprises filtrate, washing and drying to obtain a solid nucleoside product after the coupling reaction.
The deprotection treatment in the second step comprises the following steps:
a. dissolving the obtained solid nucleoside product in a solvent;
b. adding an alkaline or acidic deprotection agent, wherein the deprotection agent comprises sodium hydroxide;
c. deprotection reaction is carried out at 25 ℃ and pH value of 5 to remove the protective group of nucleoside;
d. and (3) carrying out post-reaction treatment, wherein the post-reaction treatment comprises filtrate, neutralization, washing and drying to obtain a deprotected nucleoside product.
The specific purification steps in step three are as follows:
a. dissolving the deprotected nucleoside in a solvent;
b. solvent extraction is performed through a separating funnel to separate and remove unwanted impurities;
c. distilling the extracted solution to remove the solvent;
d. and freeze-drying the solution obtained after the solvent removal to obtain a purified nucleoside product.
The above purification steps ensure the purity of the obtained nucleoside product, providing a good quality starting material for the subsequent phosphorylation reaction.
The solvent used above was methanol.
And step four, after NMN is generated, performing secondary purification treatment on the NMN, wherein the secondary purification comprises chromatographic and crystallization methods, so that the purity of the NMN is improved.
The chromatographic purification steps are as follows:
a. dissolving the generated NMN with ethanol;
b. passing the dissolved NMN liquid through a chromatographic column to separate NMN from other possible impurities using different affinities;
c. collecting the liquid flowing out of the chromatographic column, and detecting to identify a NMN-containing part;
d. the NMN-containing fraction is further freed from the solution by evaporation, resulting in purified NMN.
By purification through chromatography, NMN can be effectively separated from impurities which may be present, ensuring the purity and quality of the final product.
The crystallization and purification steps are as follows:
a. dissolving NMN obtained after chromatographic purification with ethanol;
b. slowly reducing the temperature of the solution to enable NMN to start crystallization;
c. centrifuging or filtering, and separating NMN crystals from the mother liquor;
d. washing the NMN crystals with a suitable ethanol solvent to remove impurities that may attach to the crystal surfaces;
e. and drying the washed NMN crystals to obtain NMN products with higher purity.
Purification by crystallization can further increase the purity of the NMN product and convert it to a solid form suitable for storage and use.
The prepared NMN anti-aging substance is used for delaying human aging and treating diseases caused by cell aging.
Example 1 yield data
| Project | Example 1 |
| Conversion of reaction (%) | 78 |
| Purity (%) | 98 |
| Total output (g) | 392 |
Examples
In this example, 0.8 mole of tetraacetylribose and 0.8 mole of nicotinamide are reacted in acetic acid to give the coupled product, which is then deprotected to give the deprotected nucleoside. The temperature in the coupling and deprotection treatment is 50 ℃, the pH value is 7, the deprotection agent adopts potassium hydroxide, the solvent adopts acetone, and then the obtained nucleoside is purified in ethanol to obtain the purified nucleoside. Finally, the purified nucleoside was reacted with 0.8 unit/mole of nucleoside phosphorylase to give NMN. The NMN obtained is purified by chromatography and crystallization, and the NMN product in solid form is obtained by freeze-drying.
Example 2 yield data
| Project | Example 2 |
| Conversion of reaction (%) | 82 |
| Purity (%) | 99 |
| Total output (g) | 410 |
Examples
In this example, 1 mole of tetraacetylribose and 1 mole of nicotinamide are reacted in acetic acid to give the coupled product, which is then deprotected to give the deprotected nucleoside. The temperature in the coupling and deprotection treatment is 75 ℃, the pH value is 8, the deprotection agent adopts hydrochloric acid, the solvent adopts ethyl acetate, and then the obtained nucleoside is purified in isopropanol to obtain the purified nucleoside. Finally, the purified nucleoside is reacted with 1 unit/mole of nucleoside phosphorylase to obtain NMN. The NMN obtained is purified by chromatography and crystallization, and the NMN product in solid form is obtained by freeze-drying.
Example 3 production of data
| Project | Example 3 |
| Conversion of reaction (%) | 80 |
| Purity (%) | 99.5 |
| Total output (g) | 400 |
Examples
In this example, 0.6 mole of tetraacetylribose and 0.6 mole of nicotinamide are reacted in acetic acid to give the coupled product, which is then deprotected to give the deprotected nucleoside. The temperature in the coupling and deprotection treatment is 100 ℃, the pH value is 9, the deprotection agent adopts sulfuric acid, the solvent adopts ethyl acetate, and then the obtained nucleoside is purified in acetone to obtain the purified nucleoside. Finally, the purified nucleoside was reacted with 0.6 unit/mole of nucleoside phosphorylase to give NMN. The NMN obtained is purified by chromatography and crystallization, and the NMN product in solid form is obtained by freeze-drying.
Example 4 production of data
| Project | Example 4 |
| Conversion of reaction (%) | 79 |
| Purity (%) | 98.5 |
| Total output (g) | 395 |
From the above examples and experimental data, it can be seen that the NMN anti-aging substance of example 3 has the highest purity (99.5%) and higher reaction conversion (80%) and yield of about 400 g. Therefore, embodiment 3 is the best embodiment.
Example 3 the reaction of 1 mole of tetraacetylribose with 1 mole of nicotinamide in acetic acid followed by purification of the resulting nucleoside in isopropanol and reaction with 1 unit/mole of nucleoside phosphorylase and finally purification by chromatography and crystallization. The process flow and steps of example 3 are not much different from other examples, but experimental results show that the conditions of example 3 may be more advantageous for efficient, high purity NMN production.
The following is a table of data comparing process time, energy consumption, and material costs for each example
| Items/embodiments | Example 1 | Example 2 | Example 3 | Example 4 |
| Total process time (hours) | 8 | 7.5 | 7 | 8 |
| Total energy consumption (kilowatt-hour) | 10 | 9.5 | 8.5 | 10 |
| Total material cost (Yuan) | 3000 | 2900 | 2800 | 3000 |
By comparison of the comparison tables, we can clearly see the comparison of process time, energy consumption and material cost for each example.
From the process time point of view, the process time of example 3 is the shortest, taking only 7 hours, indicating that the preparation process of example 3 is the most efficient.
From the energy consumption point of view, the total energy consumption of example 3 is the lowest, only 8.5 kwh, indicating that the preparation process of example 3 is the best energy saving.
From a material cost standpoint, example 3 has the lowest material cost, requiring only 2800 yuan, indicating that the manufacturing process of example 3 is most cost effective.
Therefore, example 3 shows superiority in terms of process time, energy consumption and material cost in terms of various performance comparisons, and can be regarded as the best example.
Compared with the traditional product:
purity of the product
The purity of the product of the traditional NMN preparation method is generally below 95%, while the purity of the product of the best embodiment (embodiment 3) of the invention can reach 99.5%, which obviously improves the quality of the product.
Production cost
The traditional method has the defects of high material and energy consumption and high cost due to the need of multiple purification steps. In the invention, the total material cost of the embodiment 3 is 2800 yuan, the total energy consumption is only 8.5 kilowatt-hours by optimizing the preparation process, and the production cost is greatly reduced compared with the traditional method.
Process time
Conventional methods require long reaction and purification times, typically over 10 hours. In the embodiment 3 of the invention, the whole preparation process only needs 7 hours, so that the production efficiency is greatly improved.
Conversion of reaction
The traditional method has low reaction conversion rate due to the fact that the reaction conditions are not easy to control. The reaction conversion rate of the embodiment 3 of the invention can reach 80 percent, which is obviously improved compared with the traditional method.
Therefore, compared with the traditional method, the preparation process of the NMN anti-aging substance has obvious advantages in the aspects of product purity, production cost, process time, reaction conversion rate and the like, and has higher practical value and wide application prospect.
The present invention is intended to embrace all such alternatives, modifications and variances which fall within the broad scope of the appended claims. Therefore, any omission, modification, equivalent replacement, improvement, etc. of the present invention should be included in the scope of the present invention.
Claims (9)
1. A process for preparing an NMN anti-ageing substance, comprising the steps of:
step one: coupling reaction is carried out on tetraacetylribose and nicotinamide by adopting a chemical method to generate nucleoside;
step two: deprotection treatment is carried out on the generated nucleoside, and a protecting group is removed;
step three: purifying the deprotected nucleoside to obtain purified nucleoside;
step four: reacting the purified nucleoside with a phosphorylase to produce NMN;
the tetraacetylribose content is 0.01-1 mol, the nicotinamide content is 0.01-1 mol, and the phosphorylase content is 0.01-1 unit/mol nucleoside.
2. The process for preparing an NMN anti-aging substance according to claim 1, wherein the coupling reaction in the step one is as follows:
a. mixing tetraacetylribose and nicotinamide in a molar ratio of 1:1 in a solvent to obtain a mixture;
b. adjusting the pH value of the mixture to be in the range of 5-9 so as to facilitate the next coupling reaction;
c. carrying out coupling reaction at 25-100 ℃ to generate nucleoside;
d. and (3) carrying out post-treatment after the reaction is finished, wherein the post-treatment comprises filtrate, washing and drying to obtain a solid nucleoside product after the coupling reaction.
3. The process for preparing an NMN anti-aging substance according to claim 1, wherein the step of deprotecting in step two is as follows:
a. dissolving the obtained solid nucleoside product in a solvent;
b. adding an alkaline or acidic deprotection agent, wherein the deprotection agent comprises sodium hydroxide, potassium hydroxide, hydrochloric acid or sulfuric acid;
c. deprotection reaction is carried out at the temperature of 25-100 ℃ and the pH value of 5-9, and the protective group of nucleoside is removed;
d. and (3) carrying out post-reaction treatment, wherein the post-reaction treatment comprises filtrate, neutralization, washing and drying to obtain a deprotected nucleoside product.
4. The process for preparing an NMN anti-aging substance according to claim 1, wherein the specific purification steps in step three are as follows:
a. dissolving the deprotected nucleoside in a solvent;
b. solvent extraction is performed through a separating funnel to separate and remove unwanted impurities;
c. distilling the extracted solution to remove the solvent;
d. and freeze-drying the solution obtained after the solvent removal to obtain a purified nucleoside product.
5. The process for preparing an NMN anti-aging substance according to any one of claims 2 to 4, wherein the solvent comprises an alcohol, a ketone or an ester, wherein the alcohol is methanol, the ketone is acetone, and the ester is ethyl acetate.
6. The process for preparing NMN anti-aging substance according to claim 1, wherein after NMN is generated in the fourth step, the NMN is subjected to a secondary purification treatment, and the secondary purification comprises chromatography and crystallization methods, so that the purity of NMN is improved.
7. The process for preparing an NMN anti-aging substance according to claim 6, wherein the chromatographic purification steps are as follows:
a. dissolving the generated NMN with ethanol;
b. passing the dissolved NMN liquid through a chromatographic column to separate NMN from other possible impurities using different affinities;
c. collecting the liquid flowing out of the chromatographic column, and detecting to identify a NMN-containing part;
d. the NMN-containing fraction is further freed from the solution by evaporation, resulting in purified NMN.
8. The process for preparing an NMN anti-aging substance according to claim 7, wherein the crystallization and purification steps are as follows:
a. dissolving NMN obtained after chromatographic purification with ethanol;
b. slowly reducing the temperature of the solution to enable NMN to start crystallization;
c. centrifuging or filtering, and separating NMN crystals from the mother liquor;
d. washing the NMN crystals with a suitable ethanol solvent to remove impurities that may attach to the crystal surfaces;
e. and drying the washed NMN crystals to obtain NMN products with higher purity.
9. The process for preparing an NMN anti-aging substance according to claim 1, wherein the prepared NMN anti-aging substance is used for delaying aging of human body and treating diseases caused by cell aging.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310789714.4A CN116751826A (en) | 2023-06-30 | 2023-06-30 | Preparation process of NMN anti-aging substance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310789714.4A CN116751826A (en) | 2023-06-30 | 2023-06-30 | Preparation process of NMN anti-aging substance |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116751826A true CN116751826A (en) | 2023-09-15 |
Family
ID=87947729
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310789714.4A Pending CN116751826A (en) | 2023-06-30 | 2023-06-30 | Preparation process of NMN anti-aging substance |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116751826A (en) |
-
2023
- 2023-06-30 CN CN202310789714.4A patent/CN116751826A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106256824B (en) | Preparation method of high-purity delafloxacin meglumine salt | |
| CN108467396B (en) | Ganciclovir preparation method | |
| CN104788461A (en) | Industrial production method applicable to citric acid tofacitinib | |
| CN102295658A (en) | Refining method of disodium phosphocreatine | |
| CN110791538A (en) | Production method suitable for synthesizing sitagliptin phosphate by enzyme method | |
| CN112979482A (en) | High-purity L-valine and preparation method and application thereof | |
| CN116751826A (en) | Preparation process of NMN anti-aging substance | |
| CN101870704B (en) | Method for purifying cefotetan acid crude products | |
| CN103601766A (en) | Fondaparinux sodium pentasaccharide intermediate and preparation method thereof | |
| CN113754626B (en) | Method for preparing fisetin by enzyme method | |
| CN113173953B (en) | Purification method of high-purity lobaplatin trihydrate for preparing antitumor drugs | |
| WO2011100670A1 (en) | Process for the preparation of scyllo-inositol | |
| CN112851543B (en) | Preparation method of methoxyamine hydrochloride and preparation method of N-methoxyacetamide | |
| EP2024377B1 (en) | Process for the preparation of n-acetyl-d-mannosamine monohydrate | |
| CN110606863B (en) | Preparation method of N-acetylneuraminic acid dihydrate | |
| CN102952165A (en) | Method for extracting L-arabinose from xylose mother liquid | |
| CN112479853A (en) | Preparation method of D-2-chloropropionyl chloride and D-2-chloropropionyl chloride | |
| CN110526950B (en) | Preparation method of alpha-five-O-acetyl mannose | |
| CN105622694A (en) | Exemestane synthesis technology | |
| CN111377840A (en) | Preparation method of R- (+) -dihydrolipoic acid | |
| CN114907228B (en) | Colchicine and magnolol compound, synthesis method thereof and application thereof in resisting new coronaviruses | |
| CN115838363B (en) | Purification method of mycophenolic acid | |
| CN112341508B (en) | Separation and purification method of ademetionine | |
| CN111454214B (en) | Synthetic method of 2-methoxy-1-pyrimidineethylamine hydrochloride | |
| CN113979886A (en) | Protected amino acid with amide side chain and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |