阅读说明：本技术 抗衰老药物组合及其用途 (Anti-aging pharmaceutical composition and application thereof ) 是由 李丹 陈芳芳 刘华英 王炳荣 廖炜程 于 2021-08-30 设计创作，主要内容包括：本发明属于营养和药物领域,涉及一种抗衰老药物组合及其用途。具体地,本发明涉及药物组合,包括：NR或其可药用盐0.5-30重量份,NMN 0.5-30重量份,以及矿物质0.01-35重量份。本发明的药物组合具有良好的抗氧化或抗衰老效果,并且具有优异的稳定性。(The invention belongs to the field of nutrition and medicine, and relates to an anti-aging medicine composition and application thereof. In particular, the present invention relates to a pharmaceutical combination comprising: 0.5-30 parts of NR or a pharmaceutically acceptable salt thereof, 0.5-30 parts of NMN, and 0.01-35 parts of mineral. The pharmaceutical composition has good antioxidant or anti-aging effect and excellent stability.)

1. A pharmaceutical combination comprising:

NR or a pharmaceutically acceptable salt thereof 0.5 to 30 parts by weight,

0.5 to 30 parts by weight of NMN, and

0.01-35 parts of mineral substance.

2. The pharmaceutical combination according to claim 1, comprising:

NR or a pharmaceutically acceptable salt thereof 0.7 to 30 parts by weight,

0.8 to 30 parts by weight of NMN, and

0.01-30 parts of mineral; or

NR or a pharmaceutically acceptable salt thereof 0.7 to 30 parts by weight,

0.8 to 30 parts by weight of NMN, and

1-30 parts of mineral substance; or

NR or a pharmaceutically acceptable salt thereof 0.5 to 25 parts by weight,

0.5 to 25 parts by weight of NMN, and

8-32 parts of mineral substances; or

NR or a pharmaceutically acceptable salt thereof 0.7 to 24 parts by weight,

0.8 to 21 parts by weight of NMN, and

10-30 parts of mineral substances; or

9 to 24 parts by weight of NR or a pharmaceutically acceptable salt thereof,

NMN 9-21 parts by weight, and

10-20 parts of mineral substances;

or

9 parts by weight of NR or a pharmaceutically acceptable salt thereof,

NMN 9 parts by weight, and

10 parts of mineral matters.

3. The pharmaceutical combination according to any one of claims 1 to 2, wherein the pharmaceutically acceptable salt is NR chloride.

4. The pharmaceutical combination according to any one of claims 1 to 3, wherein,

the weight ratio of NR or a pharmaceutically acceptable salt thereof to NMN is (1: 30) to (30: 1), preferably (1: 3) to (3: 1); more preferably 1: 1.

5. the pharmaceutical combination according to any one of claims 1 to 4, wherein

The ratio of the sum of the weight of NR or a pharmaceutically acceptable salt thereof and the weight of NMN to the weight of the mineral is (0.5: 1) to (1000: 1), (0.5: 1) to (250: 1), or (0.5: 1) to (30: 1);

preferably (0.5: 1) to (3: 1) or (0.7: 1) to (2.5: 1);

more preferably (1: 1) to (2: 1);

particularly preferably 1.8: 1.

6. the pharmaceutical combination according to any one of claims 1 to 5, wherein,

NR or a pharmaceutically acceptable salt thereof and NMN are not in direct contact with minerals;

preferably, NR or a pharmaceutically acceptable salt thereof is contained in microcapsules simultaneously and/or separately with NMN;

preferably, the microcapsules are enteric microcapsules;

preferably, the microcapsules further contain an antioxidant; preferably, the antioxidant is a water-soluble antioxidant and/or an oil-soluble antioxidant;

preferably, the microcapsule consists of a core material and at least two layers of wall materials, wherein the core material comprises NR or a pharmaceutically acceptable salt thereof and NMN, the first layer of the wall materials comprises grease with a melting point higher than 38 ℃ and covers the outer surface of the core material, and the second layer of the wall materials comprises a flowing coating material and covers the outer surface of the first layer of the wall materials;

preferably, the microcapsules comprise, in weight percent of the microcapsules: 1-30% of NR or a pharmaceutically acceptable salt thereof, 1-30% of NMN, 40-80% of grease with the melting point higher than 38 ℃, 5-15% of surfactant and 5-25% of a free-flowing coating material;

preferably, the microcapsules are prepared by a process comprising the steps of:

(1) dissolving NR or a pharmaceutically acceptable salt thereof and NMN in water to prepare a water-phase material;

(2) uniformly mixing an emulsifier and grease with a melting point higher than 38 ℃ to prepare an oil phase material;

(3) mixing the water phase material obtained in the step (1) and the oil phase material obtained in the step (2), and emulsifying to obtain a water-in-oil type emulsion;

(4) dehydrating the water-in-oil emulsion obtained in the step (3) to obtain a dehydrated water-in-oil emulsion; and

(5) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation to prepare microcapsules; preferably, the enteric layer is continuously sprayed in the fluidized bed to obtain the enteric microcapsule.

7. The pharmaceutical combination according to any one of claims 1 to 6, wherein,

the mineral comprises one or more selected from calcium, magnesium, zinc, selenium and chromium;

preferably, the source of calcium is one or more selected from calcium citrate, calcium carbonate and calcium gluconate;

preferably, the source of magnesium is one or more selected from magnesium sulfate, magnesium hydroxide and magnesium oxide;

preferably, the source of zinc is one or more selected from zinc lactate, zinc sulfate and zinc gluconate;

preferably, the source of selenium is one or more selected from sodium selenite and selenium-enriched yeast;

preferably, the source of chromium is one or more selected from chromium chloride and chromium-enriched yeast;

preferably, the minerals further comprise one or more selected from manganese (e.g. manganese gluconate), sodium, potassium, iron and copper.

8. The pharmaceutical combination according to any one of claims 1 to 7, wherein the minerals comprise, by total weight of minerals:

1200 parts by weight of calcium 800-;

preferably, comprises 1100 parts by weight of calcium 900-;

more preferably, the feed comprises 1100 parts by weight of calcium gluconate 900-;

particularly preferably, it comprises:

1000 parts by weight of calcium citrate, 150 parts by weight of magnesium sulfate, 15 parts by weight of zinc lactate, 0.1 part by weight of sodium selenite and 1 part by weight of chromium chloride; or

1000 parts of calcium carbonate, 1500 parts of magnesium hydroxide, 150 parts of zinc sulfate, 0.1 part of selenium-enriched yeast and 0.1 part of chromium-enriched yeast; or

1000 parts of calcium gluconate, 150 parts of magnesium oxide, 15 parts of zinc gluconate, 0.1 part of selenium-enriched yeast, 0.1 part of chromium-enriched yeast and 2.5 parts of manganese gluconate.

9. The pharmaceutical combination according to any one of claims 1 to 8, which is a pharmaceutical composition;

optionally, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients;

preferably, the pharmaceutical composition is a capsule, a tablet or a granule.

10. The pharmaceutical combination according to any one of claims 1 to 8, which is a combination product comprising a first product and a second product in separate packages,

wherein the content of the first and second substances,

the first product comprises NR, or a pharmaceutically acceptable salt thereof, and NMN;

the second product comprises a mineral;

preferably, the first product and the second product further independently comprise one or more pharmaceutically acceptable excipients;

preferably, the first product and the second product are independently capsules, tablets or granules;

preferably, the combination product further comprises product instructions.

11. Use of a pharmaceutical combination according to any one of claims 1 to 10 for the manufacture of a medicament for anti-oxidant or anti-aging.

12. The pharmaceutical combination according to any one of claims 1 to 10 for use in anti-oxidation or anti-aging.

13. A method of anti-oxidation or anti-aging comprising the step of administering to a subject in need thereof an effective amount of the pharmaceutical combination according to any one of claims 1 to 10.

Technical Field

The invention belongs to the field of nutrition and medicine, and relates to an anti-aging medicine composition and application thereof.

Background

Aging is an activity accompanying the life process, and is continuously generated from fertilized eggs to death, and the characteristics of aging are obviously shown only by a certain stage. The physiological changes in the aging process of human body are mainly reflected in the loss of organism tissue cells and constitutional substances, the slowing of organism metabolic rate and the hypofunction of organism and organs. Senescence is inevitable, but delaying senescence is possible.

Mitochondria in human cells are the prime mover for maintaining young humans, and with age, mitochondria become less viable, and one of the most significant changes in body cells is a decrease in NAD + levels, linking the decrease in NAD + levels to aging-related diseases. NAD + is known as nicotinamide adenine dinucleotide, coenzyme I for short, is a coenzyme of dehydrogenase which is important in human body, such as alcohol dehydrogenase, used for oxidizing alcohol, plays an irreplaceable role in glycolysis, gluconeogenesis, tricarboxylic acid cycle and respiratory chain, an intermediate product can transfer the removed hydrogen to NAD to enable the NADH to become NADH, and the NADH can be used as a hydrogen carrier, synthesizes ATP in the respiratory chain through a chemical osmotic coupling mode, is a reducing agent and an electron donor, participates in different cellular processes, including oxidation and antioxidation systems, participates in mitochondrial oxidative metabolic processes, serves as a signal molecule for ensuring cell survival and regulating energy metabolism, cell repair and circadian rhythm, and has a regulating effect on physiological functions of various enzymes in cells in the form of the coenzyme. When the level of NAD + is decreased, it causes a series of processes in the cell to malfunction, resulting in an increase in the level of inflammation in the cell, mitochondrial dysfunction, and a series of NAD + -dependent enzyme dysfunction, metabolic abnormalities in the cell, and DNA damage, etc., which eventually causes the cell to go to senescence.

NAD + has an excessively high molecular weight, and is difficult to permeate cell membranes to enter the interior of cells, and cannot be taken into cells by oral administration, and depends mainly on cell synthesis in vivo. NAD + precursor species are: tryptophan, nicotinic acid, nicotinamide riboside, reduced nicotinamide riboside, nicotinamide mononucleotide, trigonelline, nicotinic acid mononucleotide, nicotinic acid riboside, and the like. It was found that niacin, nicotinamide and tryptophan all have limited intake levels, that the combination of niacin and GPR109A leads to severe flushing of the patient, and that excessive intake of tryptophan and nicotinamide also has side effects, for example, nicotinamide may cause inhibition of deacetylase. Among the numerous complementary NAD + intermediates, nicotinamide ribose and nicotinamide mononucleotide are the most effective and are converted to NAD + via a salvage synthesis pathway.

Nicotinamide Ribose (NR) is a precursor of an important coenzyme NAD +, plays an important role in the generation of human cell energy, participates in the synthesis of NAD + in cells, can obviously improve the cell activity, particularly the activity of aging cells, has obvious improvement effect on the aspects of metabolism, immunity, brain function, cardiovascular function and the like of human bodies, and can lead the cells in the bodies to be in a new active state by integrally improving the functions of the human bodies so as to achieve the aim of delaying aging.

Nicotinamide mononucleotide is a naturally occurring biologically active nucleotide, with 2 irregularly occurring forms, i.e., the α and β, β isomers are the active forms of nicotinamide mononucleotide. Beta-nicotinamide mononucleotide (hereinafter NMN), one of the key precursors of NAD +. Research shows that the consumption of NMN can effectively improve the content of NAD + in vivo and obviously inhibit metabolism caused by aging, so that the NMN becomes a 'non-senility medicine'. In mammals, NMN is produced by nicotinamide under catalysis of Nampt, followed by NMN producing NAD + under catalysis of nicotinamide mononucleotide adenyl transferase. Researches show that the NMN in the organism can be adjusted to have better treatment and repair effects on cardiovascular and cerebrovascular diseases, neurodegenerative diseases, aging degenerative diseases and the like.

The mineral is one of seven essential nutrients for human body. The essential minerals for human body include calcium, phosphorus, magnesium, potassium, sodium, sulfur and chlorine 7 kinds, the content of which accounts for more than 0.01% of human body or the dietary intake is more than 100mg/d, and the minerals are called macroelements; 8 kinds of iron, zinc, copper, cobalt, molybdenum, selenium, iodine and chromium are necessary microelements, and the microelements refer to mineral substances with the content of less than 0.01 percent of the human body or the dietary intake of less than 100 mg/d; 5 kinds of manganese, silicon, nickel, boron and vanadium are possible trace elements necessary for human body; some trace elements are potentially toxic, and may cause diseases or injuries to human bodies once being excessively ingested, but are possible essential trace elements to human bodies at low doses, and the trace elements mainly comprise: fluorine, lead, mercury, aluminum, arsenic, tin, lithium, cadmium, and the like. Mineral substances are the constituent elements of the human body enzyme system, the mineral substances necessary for the human body are the activity centers of a plurality of enzymes of the human body, more than 70 percent of thousands of enzymes found in the human body contain the mineral substances or are related to the mineral substances, the mineral substances can only be taken from the outside, the human body can not generate the mineral substances by itself, and the mineral substances have the functions of improving the oxidation resistance of the human body, killing free radicals and resisting aging.

In the prior art, no research and report on the simultaneous use of NR, NMN and minerals are found.

Disclosure of Invention

The present invention has been made in an intensive study and inventive work, and surprisingly found that a combination of NR, NMN and a mineral produces an unexpected synergistic effect in oxidation resistance and/or aging resistance, that is, the effect of taking the combination of the three is more excellent than the sum of the effects of the mineral alone or the (NR + NMN) alone. It has further been found in the present invention that NR or NMN, when formulated into conventional dosage forms, becomes abnormally unstable in the presence of minerals, i.e. minerals have been found to accelerate NR and NMN degradation. Since NR and NMN are quite expensive, this loss due to degradation is unacceptable. The following invention is thus provided:

one aspect of the present invention relates to a pharmaceutical combination comprising:

NR or a pharmaceutically acceptable salt thereof 0.5 to 30 parts by weight,

0.5 to 30 parts by weight of NMN, and

0.01-35 parts of mineral substance.

In some embodiments of the invention, the pharmaceutical composition comprises:

NR or a pharmaceutically acceptable salt thereof 0.7 to 30 parts by weight,

0.8 to 30 parts by weight of NMN, and

0.01-30 parts of mineral.

In some embodiments of the invention, the pharmaceutical composition comprises:

NR or a pharmaceutically acceptable salt thereof 0.7 to 30 parts by weight,

0.8 to 30 parts by weight of NMN, and

1-30 parts of mineral substance.

In some embodiments of the invention, the pharmaceutical composition comprises:

NR or a pharmaceutically acceptable salt thereof 0.5 to 25 parts by weight,

0.5 to 25 parts by weight of NMN, and

8-32 parts of mineral substance.

In some embodiments of the invention, the pharmaceutical composition comprises:

NR or a pharmaceutically acceptable salt thereof 0.7 to 25 parts by weight,

0.8 to 25 parts by weight of NMN, and

8-32 parts of mineral substance.

In some embodiments of the invention, the pharmaceutical composition comprises:

NR or a pharmaceutically acceptable salt thereof 0.7 to 24 parts by weight,

0.8 to 21 parts by weight of NMN, and

10-30 parts of mineral matters.

In some embodiments of the invention, the pharmaceutical composition comprises:

9 to 24 parts by weight of NR or a pharmaceutically acceptable salt thereof,

NMN 9-21 parts by weight, and

10-20 parts of mineral.

In some embodiments of the invention, the pharmaceutical composition comprises:

9 parts by weight of NR or a pharmaceutically acceptable salt thereof,

NMN 9 parts by weight, and

10 parts of mineral matters.

In some embodiments of the invention, the pharmaceutical combination is wherein the pharmaceutically acceptable salt is NR chloride.

In some embodiments of the invention, the pharmaceutical combination is 0.5, 0.7, 9, 24 or 25 parts by weight of NR or a pharmaceutically acceptable salt thereof.

In some embodiments of the invention, the pharmaceutical combination wherein the NMN is 0.8, 9, 21 or 25 parts by weight.

In some embodiments of the invention, the pharmaceutical composition is a composition comprising 8 parts by weight, 10 parts by weight, 20 parts by weight, 30 parts by weight, or 32 parts by weight of a mineral.

In some embodiments of the invention, the pharmaceutical combination, wherein,

the weight ratio of NR or a pharmaceutically acceptable salt thereof to NMN is (1: 30) to (30: 1), preferably (1: 10) to (10: 1), (1: 5) to (5: 1), (1: 4) to (4: 1), (1: 3) to (3: 1), (1: 2) to (2: 1), (1: 1.5) to (1.5: 1); more preferably 1: 1.

in some embodiments of the invention, the pharmaceutical combination, wherein,

the ratio of the sum of the weight of NR or a pharmaceutically acceptable salt thereof and the weight of NMN to the weight of the mineral is (0.5: 1) to (1000: 1), (0.5: 1) to (250: 1), or (0.5: 1) to (30: 1); preferably (0.5: 1) to (3: 1); more preferably (0.7: 1) to (2.5: 1), (1: 1) to (2: 1), 1.8: 1. 0.7: 1. 1.5: 1 or 1.25: 1.

in some embodiments of the invention, the pharmaceutical combination, wherein,

NR or a pharmaceutically acceptable salt thereof and NMN are not in direct contact with minerals;

preferably, NR or a pharmaceutically acceptable salt thereof is contained in microcapsules simultaneously and/or separately with NMN;

preferably, the microcapsules are enteric microcapsules;

preferably, the microcapsules further contain an antioxidant; preferably, the antioxidant is a water-soluble antioxidant and/or an oil-soluble antioxidant;

preferably, the microcapsule consists of a core material and at least two layers of wall materials, wherein the core material comprises NR or a pharmaceutically acceptable salt thereof and NMN, the first layer of the wall materials comprises grease with a melting point higher than 38 ℃ and covers the outer surface of the core material, and the second layer of the wall materials comprises a flowing coating material and covers the outer surface of the first layer of the wall materials;

preferably, the microcapsules are prepared by a process comprising the steps of:

(1) dissolving NR or a pharmaceutically acceptable salt thereof and NMN in water to prepare a water-phase material;

(2) uniformly mixing an emulsifier and grease with a melting point higher than 38 ℃ to prepare an oil phase material;

(3) mixing the water phase material obtained in the step (1) and the oil phase material obtained in the step (2), and emulsifying to obtain a water-in-oil type emulsion;

(4) dehydrating the water-in-oil emulsion obtained in the step (3) to obtain a dehydrated water-in-oil emulsion; and

(5) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation to prepare microcapsules; preferably, the enteric layer is continuously sprayed in the fluidized bed to obtain the enteric microcapsule.

In the step (2), the emulsifier and the grease may be mixed first, then melted and stirred uniformly, or the grease may be melted first and then mixed uniformly with the emulsifier.

The NR and NMN of the invention take the grease with the melting point higher than 38 ℃ and the flow-dispersing coating material as the embedding wall material of the microcapsule, have better air isolation effect compared with the traditional water-soluble colloid, can better resist high-temperature and high-humidity environment, grease environment and machining environment, and greatly improve the stability of the composition.

In one embodiment of the present invention, the pharmaceutical composition, wherein the microcapsule is an enteric microcapsule, and can be safely released in the intestinal tract through gastric juice, so as to avoid the destruction of NR and NMN by gastric acid, thereby improving the absorption and utilization rate.

In some embodiments of the invention, the pharmaceutical combination, wherein,

the mineral comprises one or more selected from calcium, magnesium, zinc, selenium and chromium;

preferably, the source of calcium is one or more selected from calcium oxide, calcium phosphate, calcium lactate, calcium citrate, calcium carbonate and calcium gluconate;

preferably, the source of magnesium is one or more selected from magnesium sulfate, magnesium hydroxide and magnesium oxide;

preferably, the source of zinc is one or more selected from the group consisting of zinc lactate, zinc chloride, zinc sulfate and zinc gluconate;

preferably, the source of selenium is one or more selected from sodium selenite and selenium-enriched yeast;

preferably, the source of chromium is one or more selected from chromium chloride, chromium pyridine, and chromium-enriched yeast;

preferably, the minerals further comprise one or more selected from manganese (e.g. manganese gluconate), sodium, potassium, iron and copper.

In some embodiments of the invention, the pharmaceutical composition comprises, in combination, at least one mineral selected from the group consisting of:

1200 parts by weight of calcium 800-;

preferably, comprises 1100 parts by weight of calcium 900-;

more preferably, the feed comprises 1100 parts by weight of calcium gluconate 900-;

particularly preferably, it comprises:

1000 parts by weight of calcium citrate, 150 parts by weight of magnesium sulfate, 15 parts by weight of zinc lactate, 0.1 part by weight of sodium selenite and 1 part by weight of chromium chloride; or

1000 parts of calcium carbonate, 1500 parts of magnesium hydroxide, 150 parts of zinc sulfate, 0.1 part of selenium-enriched yeast and 0.1 part of chromium-enriched yeast; or

1000 parts of calcium gluconate, 150 parts of magnesium oxide, 15 parts of zinc gluconate, 0.1 part of selenium-enriched yeast, 0.1 part of chromium-enriched yeast and 2.5 parts of manganese gluconate.

In some embodiments of the invention, the pharmaceutical combination is a pharmaceutical composition;

optionally, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients;

preferably, the pharmaceutical composition is a capsule, a tablet or a granule.

In some embodiments of the invention, the pharmaceutical combination, which is a combination product, comprises a first product and a second product in separate packages,

wherein the content of the first and second substances,

the first product comprises NR, or a pharmaceutically acceptable salt thereof, and NMN;

the second product comprises a mineral;

preferably, the first product and the second product further independently comprise one or more pharmaceutically acceptable excipients;

preferably, the first product and the second product are independently capsules, tablets or granules;

preferably, the combination product further comprises product instructions.

The pharmaceutical combination according to any one of the present invention for use in anti-oxidant or anti-aging.

Another aspect of the present invention relates to the use of a pharmaceutical combination according to any one of the present invention for the manufacture of a medicament for anti-oxidation or anti-aging.

Yet another aspect of the present invention relates to a method of anti-oxidation or anti-aging comprising the step of administering to a subject in need thereof an effective amount of a pharmaceutical combination according to any one of the present invention.

In the present invention, with respect to the weight or parts by weight of the mineral, calcium is calculated by the weight or parts by weight of the calcium salt, not by the weight of the calcium element, if not specifically stated. The weight or parts by weight of other minerals such as calcium, zinc, selenium, chromium, manganese, etc. are similarly understood.

In the present invention, for the sake of brevity, pharmaceutically acceptable salts include not only pharmaceutically acceptable salts but also nutritionally acceptable salts.

In the present invention, the mineral includes inorganic minerals and/or organic minerals, the inorganic minerals being inorganic salts, which are any pharmaceutically or nutritionally acceptable salts, such as carbonates, nitrates, halides, oxides, stearates, sulfates, phosphates, pyrophosphates, bicarbonates or dihydrogenphosphates; the organic mineral is the product of combining mineral and organic molecule, such as selenium-enriched yeast, chromium-enriched yeast, gluconate, citrate, lactate, etc.

In some embodiments of the present invention, the microcapsules comprise, in weight percent (based on the total weight of the microcapsules): 1-30% of NR or a pharmaceutically acceptable salt thereof, 1-30% of NMN, 40-80% of grease with the melting point higher than 38 ℃, 5-15% of surfactant and 5-25% of a free-flowing coating material.

In the present invention, the NR pharmaceutically or nutritionally acceptable salt is selected from at least one of chloride, fluoride, oxide, bromide, rosasite, formate, acetate, ascorbate, benzoate, carbonate, citrate, carbamate, formate, gluconate, lactate, methyl bromide, methyl sulfate, nitrate, phosphate, diphosphate, succinate, sulfate and difluoroacetate.

In the present invention, the fat or oil having a melting point of higher than 38 ℃ is at least one of wax, higher fatty acid, and hardened oil.

In the present invention, the wax may be animal wax, vegetable wax, mineral wax and synthetic wax, such as beeswax, insect wax, chinese wax, spermaceti wax, wool wax, palm wax, coggygria wax, coconut wax, food grade paraffin wax, ozokerite wax, microcrystalline wax, etc.

In the present invention, the higher fatty acid may be a saturated or unsaturated fatty acid having from C16 to C24, such as stearic acid, palmitic acid, pearlescent acid, arachidic acid, and behenic acid.

In the present invention, the hardened oil may be hydrogenated cottonseed oil, hydrogenated soybean oil, or the like.

In the invention, the surfactant is at least one selected from tween, span, sucrose fatty acid ester, lecithin, polyoxyethylene fatty acid ester and polyglycerol fatty acid ester.

In the invention, the free-flowing coating material is selected from at least one of silicon dioxide, calcium phosphate, calcium hydrophosphate, calcium carbonate and starch; the starch can be corn starch, cassava starch, potato starch, sweet potato starch, etc.

Further, the microcapsule can also comprise an antioxidant, wherein the antioxidant accounts for 0.5 to 2 percent of the weight of the microcapsule; the antioxidant can be a water-soluble antioxidant and/or an oil-soluble antioxidant, the water-soluble antioxidant is mainly distributed in the core material of the microcapsule, the oil-soluble antioxidant is mainly distributed in the wall material of the microcapsule, and the water-soluble antioxidant and the oil-soluble antioxidant can exist at the same time or only contain one. Further, the antioxidant is at least one selected from coenzyme Q10, sodium erythorbate, ascorbyl palmitate, tea polyphenol, tocopherol, lipoic acid, BHT, BHA, TBHQ and rosemary extract.

In the invention, the microcapsule also comprises an enteric layer, wherein the enteric layer accounts for 2 to 20 percent of the weight of the microcapsule; the main component of the enteric layer is polyacrylic resin, and the main component comprises at least one of polyacrylic resin I, polyacrylic resin II, polyacrylic resin III, polyacrylic resin IV, L30D-55, L100D-55, E100/EPO, E12.5, L100, L12.5, S100, S12.5 and FS 30D.

The enteric layer further comprises an anti-sticking agent and a plasticizer, the anti-sticking agent mainly comprises one or more of talcum powder, diatomite and silicon dioxide, and the plasticizer mainly comprises one or more of polyethylene glycol, triethyl citrate, triacetin, glycerol and propylene glycol.

In some embodiments of the invention, a method of making a microcapsule comprises:

(1) dissolving NR, NMN and water-soluble antioxidant (if any) in water to obtain a water-phase material;

(2) mixing surfactant, oil-soluble antioxidant (if any) and melted grease with melting point higher than 38 ℃ to obtain oil phase material;

(3) mixing the water phase material and the oil phase material, and emulsifying to obtain a water-in-oil type emulsion;

(4) vacuum dewatering the water-in-oil emulsion to obtain dewatered water-in-oil emulsion;

(5) spraying the dewatered water-in-oil emulsion into a fluidized bed for spray cooling granulation and sieving to obtain the product; wherein the fluidized bed is distributed with fluidized fluid coating material.

Further, in the above step (1), the amount of water is an amount which is just enough to dissolve NR and NMN.

Further, in the step (3), the emulsification method is at least one of a high-speed shearing method, a high-pressure homogenization method, a cavitation emulsification method, and a micro-jet method.

Further, in the step (4), the vacuum degree of the vacuum dewatering is-0.05 MPa to-0.1 MPa.

Further, in the steps (2) to (4), the temperature of the material is controlled to be higher than 38 ℃.

Further, in the step (5), the temperature of the material is controlled to be less than 38 ℃, preferably less than 30 ℃.

In one embodiment of the present invention, after the step (5) above, the enteric layer is sprayed by a fluidized bed coater, which controls the temperature of the material to be kept below 38 ℃.

In the present invention, the term "microcapsule" refers to a small particle containing an active ingredient or core material surrounded by a coating or shell.

In the present invention, the terms "first" (e.g., first product) and "second" (e.g., second product) are used for distinguishing between or for clarity of presentation and do not have a typical sequential meaning unless otherwise indicated.

Advantageous effects of the invention

The invention achieves one or more of the following technical effects:

(1) good antioxidant effect, such as obviously reducing the MDA content in the tissue and increasing the SOD content;

(2) good anti-aging or anti-aging effect;

(3) the combination of NR or a pharmaceutically acceptable salt thereof and NMN (NR or a pharmaceutically acceptable salt thereof + NMN) has a synergistic effect with minerals; the effect is better than that of only NR or the pharmaceutically acceptable salt thereof plus NMN or only mineral.

(4) Good stability;

(5) higher absorption rate.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

In the following examples, the NMN content was measured by hplc, wherein mobile phase a: 50mmol/L potassium dihydrogen phosphate, mobile phase B: acetonitrile; a chromatographic column: agilent Zorbax SB-AQ, 250X 4.6 mm; a detector: UV 254 nm; column temperature: 25 ℃; flow rate: 1.0 mL/min; sample introduction amount: 5 mu L of the solution; gradient elution conditions: time/min (0, 4, 5, 6, 10), mobile phase a/vt% (96, 80, 96), mobile phase B/vt% (4, 20, 4). The content of NR was detected by a high performance liquid chromatograph using a C18 column (250 mm. times.4.6 mm,5 μm) as a chromatographic column, a methanol-buffer (pH 3.5 containing 0.1% sodium heptanesulfonate, 20:80, v/v) as a mobile phase, a flow rate of 1mL/min, isocratic elution, a column temperature of 30 ℃ and a detection wavelength of 261 nm.

Example 1: preparation of microcapsules containing NR and NMN

(1) Dissolving 1Kg of NR chloride and 30Kg of NMN in 16L of water to obtain a water phase material;

(2) uniformly mixing 10Kg of tween-40, 5Kg of span-60 and 45Kg of molten beewax to obtain an oil phase material;

(3) adding the water phase material obtained in the step (1) into the oil phase material obtained in the step (2), and carrying out high-speed shearing emulsification for 30min at the shearing speed of 9000rpm at the temperature of 68 ℃ to obtain a water-in-oil type emulsion;

(4) gradually vacuumizing the water-in-oil emulsion obtained in the step (3) at the shearing speed of 5000rpm at 68 ℃ to start dehydration until the vacuum degree reaches-0.1 MPa and the moisture is basically and completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 9Kg of fluidized silicon dioxide is distributed in the fluidized bed, and in the operation process, the temperature of the material is controlled to be kept at 25 ℃, and sieving is carried out, so that 99.99Kg of microcapsules containing NR and NMN are obtained. By detection, the content of NR is 1.03%, and the content of NMN is 30.15%.

Example 2: preparation of microcapsules containing NR and NMN

(1) Dissolving 30Kg of NR chloride and 1Kg of NMN in 16L of water to obtain a water phase material;

(2) uniformly mixing 14Kg of triglycerol monostearate and 50Kg of molten palm wax to obtain an oil phase material;

(3) adding the water phase material obtained in the step (1) into the oil phase material obtained in the step (2), carrying out high-speed shearing emulsification at 86 ℃ and a shearing speed of 10000rpm for 15min, and carrying out high-pressure homogenization at 45MPa for 3 times to obtain a water-in-oil emulsion;

(4) gradually vacuumizing the water-in-oil emulsion obtained in the step (3) at 86 ℃ and a shearing speed of 6000rpm to start dehydration until the vacuum degree reaches-0.1 MPa and the moisture is basically and completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 5Kg of fluidized calcium phosphate is distributed in the fluidized bed, and in the operation process, the temperature of the material is controlled to be kept at 25 ℃, and sieving is carried out, so that 99.95Kg of microcapsules containing NR and NMN are obtained. By detection, the content of NR is 30.22%, and the content of NMN is 1.13%.

Example 3: preparation of enteric microcapsules containing NR and NMN

(1) Dissolving 10Kg of NR chloride and 10Kg of NMN in 10L of water to obtain a water phase material;

(2) 2Kg of lecithin, 8Kg of span-60 and 45Kg of molten stearic acid are mixed uniformly to obtain an oil phase material;

(3) simultaneously introducing the water phase material obtained in the step (1) and the oil phase material obtained in the step (2) into a multistage serial cavitation emulsifier for emulsification, wherein the temperature in the cavitation emulsifier is 72 ℃ and the pressure in the cavitation emulsifier is 300MPa to obtain a water-in-oil type emulsion;

(4) pumping the water-in-oil emulsion obtained in the step (3) into a wiped film evaporator, and carrying out continuous vacuum wiped film dehydration at 75 ℃ and under the pressure of-0.08 MPa until the water is basically completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 10Kg of fluidized corn starch is distributed in the fluidized bed, and in the operation process, the temperature of the material is controlled to be kept at 25 ℃ to obtain the microcapsule containing NR and NMN. Spraying 15kg of enteric layer in fluidized bed, wherein the formulation of the enteric layer is 40% (w/w) polyacrylic resin III, 45% (w/w) L30D-55, 5% (w/w) glyceryl triacetate and 10% (w/w) talcum powder, the solvent is 80% (w/w) ethanol water solution, and the material temperature is controlled to be 30 ℃ during the operation process. Sieving to obtain 99.90Kg enteric microcapsule containing NR and NMN. By detection, the content of NR is 10.06%, and the content of NMN is 10.24%.

Example 4: preparation of microcapsules containing NR and NMN

(1) Dissolving 15Kg of NR, 15Kg of NMN and 0.6Kg of sodium erythorbate in 16L of water to obtain a water phase material;

(2) uniformly mixing 5Kg of PEG-40 hydrogenated castor oil, 0.4Kg of tocopherol and 59Kg of molten palmitic acid to obtain an oil phase material;

(3) beating the water phase material obtained in the step (1) and the oil phase material obtained in the step (2) into a micro-jet emulsifier for continuous emulsification, wherein the emulsification temperature is 65 ℃, and obtaining a water-in-oil type emulsion;

(4) pumping the water-in-oil emulsion obtained in the step (3) into a wiped film evaporator, and carrying out continuous vacuum wiped film dehydration at 65 ℃ and under the condition of-0.1 MPa until the moisture is basically completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 5Kg of fluidized cassava starch is distributed in the fluidized bed, and in the operation process, controlling the temperature of the materials to be kept at 25 ℃, and sieving to obtain 99.70Kg of microcapsules containing NR and NMN. By detection, the content of NR is 14.93%, and the content of NMN is 15.01%.

Example 5: preparation of microcapsules containing NR and NMN

(1) Dissolving 20Kg of NR, 5Kg of NMN and 0.3Kg of tea polyphenol in 13L of water to obtain a water phase material;

(2) uniformly mixing 10Kg of sucrose ester SE-15, 0.2Kg of ascorbyl palmitate and 40Kg of molten hydrogenated cottonseed oil to obtain an oil phase material;

(3) adding the water phase material obtained in the step (1) into the oil phase material obtained in the step (2), and carrying out high-speed shearing emulsification at the shearing speed of 12000rpm at the temperature of 75 ℃ for 20min to obtain a water-in-oil type emulsion;

(4) gradually vacuumizing the water-in-oil emulsion obtained in the step (3) at the shearing speed of 6000rpm at 75 ℃ to start dehydration until the vacuum degree reaches-0.1 MPa and the moisture is basically and completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 24.5Kg of fluidized calcium hydrophosphate is distributed in the fluidized bed, and in the operation process, controlling the temperature of the materials to be kept at 25 ℃, and sieving to obtain 99.96Kg of microcapsules containing NR and NMN. By detection, the content of NR is 19.67%, and the content of NMN is 5.16%.

Example 6: preparation of microcapsules containing NR and NMN

(1) Dissolving 50Kg of NR and 50Kg of NMN in 50L of water to obtain a water phase material;

(2) uniformly mixing 50Kg of span-20, 20Kg of coenzyme Q10 and 780Kg of melted hydrogenated soybean oil to obtain an oil phase material;

(3) adding the water phase material obtained in the step (1) into the oil phase material obtained in the step (2), and carrying out high-speed shearing emulsification for 30min at the shearing speed of 9000rpm at the temperature of 75 ℃ to obtain a water-in-oil type emulsion;

(4) gradually vacuumizing the water-in-oil emulsion obtained in the step (3) at the temperature of 75 ℃ and the shearing speed of 5000rpm to start dehydration until the vacuum degree reaches-0.09 MPa and the moisture is basically and completely removed to obtain the dehydrated water-in-oil emulsion;

(5) and (3) spraying the water-in-oil emulsion obtained in the step (4) into a fluidized bed for spray cooling granulation, wherein 50Kg of fluidized silicon dioxide and corn starch (the mass ratio of the silicon dioxide to the corn starch is 1: 1) are distributed in the fluidized bed, and in the operation process, the temperature of the materials is kept at 25 ℃, and sieving is carried out, so that 998.36Kg of microcapsules containing NR and NMN are obtained. By detection, the content of NR is 4.93%, and the content of NMN is 5.06%.

Example 7-example 18: preparation of pharmaceutical compositions

The mineral substances, the microcapsules containing NR and NMN are accurately weighed according to the following table 1, and are fully and uniformly mixed to obtain the medicinal composition.

TABLE 1

Example 19: preparing vegetarian soft capsule

(1) Mixing 15 parts of hydroxypropyl methylcellulose, 10 parts of polyethylene glycol, 0.5 part of citric acid, 5 parts of ethanol and 25 parts of water to obtain a premixed solution;

(2) mixing the premixed solution with 5 parts of chitosan at 90 ℃ to obtain a glue solution;

(3) carrying out exhaust treatment on the glue solution under a vacuum condition;

(4) taking 150 parts of the pharmaceutical composition of example 7 as the content of the soft capsule;

(5) pressing the content and the exhausted glue solution into pills by using an automatic rotary capsule making machine, then blowing for shaping at 20 ℃ in a net machine, shaping after shaping for 4 hours, washing off an oil layer on the surface of the soft capsule in a pill washing machine by using ethanol, and blowing for 6 hours in the net machine to obtain the dried soft capsule;

(6) picking out the dried soft capsule, and picking out big pill, small pill, special-shaped pill, obvious screen printing pill, leaking pill, shriveled pill, thin-wall pill, bubble pill, etc. to obtain vegetarian soft capsule.

Example 20: preparation of tablets

The tablet formula comprises: as in table 2 below.

TABLE 2

The preparation method comprises the following steps: weighing the other components except the sodium stearyl fumarate according to the proportion of the formula, sieving, transferring into a mixing barrel, mixing at 15rpm for 30min, adding the sodium stearyl fumarate, mixing for 5min, and tabletting to obtain tablets with the hardness of 35N-50N.

Experimental example 1: stability test

1. Experimental sample

Sample 1: the pharmaceutical combination prepared in example 15;

sample 1-1: the microcapsules in example 15 were replaced with non-embedded NR and NMN starting materials, i.e. the composition of samples 1-1 was: 10Kg of mineral (the formula is the same as that in example 15), 9Kg of NR chloride and 9Kg of NMN, and the above substances are accurately weighed and fully and uniformly mixed to obtain the product.

Samples 1 to 2: the microcapsules of example 15 were replaced with those prepared by the following conventional technique:

dissolving 10Kg of NR chloride, 10Kg of NMN and 45Kg of sodium starch octenyl succinate in 55L of water to obtain a water phase material; 2Kg of lecithin and 8Kg of span-60 are mixed uniformly to obtain an oil phase material; and simultaneously introducing the obtained water phase material and the oil phase material into a multistage serial cavitation emulsifier for emulsification, wherein the temperature in the cavitation emulsifier is 72 ℃, and the pressure in the cavitation emulsifier is 300MPa to obtain emulsion, spraying the emulsion into a fluidized bed for adsorption granulation, distributing 10Kg of fluidized corn starch in the fluidized bed to obtain microcapsules with corn starch embedded on the surface, and carrying out fluidized drying at 60 ℃ to obtain the microcapsules containing NR and NMN. Continuously spraying 15kg of enteric layer in a fluidized bed, wherein the formulation of the enteric layer is 40% (w/w) of polyacrylic resin III, 45% (w/w) of L30D-55, 5% (w/w) of glyceryl triacetate and 10% (w/w) of talcum powder, the used solvent is 80% (w/w) of ethanol water solution, and in the operation process, the material temperature is controlled to be kept at 30 ℃, and sieving is carried out, thus obtaining the enteric coating.

That is, the compositions of samples 1-2 were: mineral 10Kg (same formula as in example 15), and 90Kg of microcapsules prepared by the above method. Accurately weighing the above materials, and mixing completely.

Samples 1 to 3: 9Kg of NR chloride and 9Kg of NMN, accurately weighing the above substances, and fully and uniformly mixing to obtain the product.

2. Experimental methods

Samples were individually filled into sealed brown vials and stability tested for long term (25 ℃, 60% RH) and accelerated conditions (40 ℃, 75% RH) according to guidelines for formulation stability experiments in chinese pharmacopoeia, 2015, respectively.

Retention rate is content at end of storage/initial content 100%.

3. Results of the experiment

As shown in table 3.

TABLE 3

From the experimental results of samples 1-1 and 1-3, it can be seen that minerals negatively affect the stability of NR and NMN, and that minerals accelerate the degradation of NR and NMN.

It can be seen from the experimental results of sample 1, sample 1-2 and sample 1-1 that the stability of NR and NMN can be greatly improved by microencapsulating NR and NMN and then mixing NR and NMN with mineral substances.

From the experimental results of samples 1 and 1-2, it can be seen that the microcapsule prepared by the present invention has better stability than the microcapsule prepared by the conventional method in the prior art, and the stability of the composition can be significantly improved compared with the conventional water-soluble colloid. The inventors speculate that the microcapsules prepared according to the invention can better insulate air and/or better withstand high temperature and high humidity environments.

Experimental example 2: anti-aging Effect test

Malondialdehyde (MDA) is a lipid peroxidation product, is closely related to the production of free radicals in the body and the lipid peroxidation degree of cells of body tissues, can reflect the oxidative stress damage state of body tissues, and is one of the important indexes for evaluating aging. Superoxide dismutase (SOD) is a key enzyme for eliminating oxygen free radicals in vivo, and the activity of the SOD can reflect the antioxidant capacity of tissues and is also an important index for evaluating the aging of organisms.

(1) Modeling, drug administration and experimental grouping

Molding: 140 Wistar rats with 120.36g +/-2.35 g weight and half male and female parts are randomly divided into 14 groups of 10 rats. Rats in each group except the blank control group were injected with 120mg/kg of 5% D-galactose subcutaneously in the neck every day to cause aging model.

Administration: continuous gavage administration was started for 30 days from day 13 of molding, and the administration volume of each group was 2 mL. Fasting is not forbidden for 24h after each administration, eyeground blood is taken, the eyeground blood is kept stand at room temperature until the blood is completely coagulated, and then centrifugation is carried out to separate serum for later use. Then, the rat is killed by a dislocation method, the liver and brain tissues are quickly separated, 10% tissue homogenate is prepared after the sodium chloride solution is washed, and the supernatant of the tissue homogenate is obtained by centrifugation and separation for later use. The SOD activity and MDA content of serum, liver and brain tissue are respectively measured by xanthine oxidase method and thiobarbituric acid spectrophotometry.

Grouping experiments:

a. blank control group: physiological saline;

b. model control group: physiological saline;

c. example 15 drug group: the composition of example 15 was mixed with physiological saline to make a 2.0g/mL suspension (i.e., NR at 0.18g/mL, NMN at 0.18g/mL, minerals at 0.2 g/mL);

d. comparative group 1: microcapsules the same as in example 15, but without the addition of minerals, i.e. microcapsules only and no minerals, were made up in a 1.8g/mL suspension with physiological saline (i.e. NR and NMN concentrations the same as in group c);

e. comparative group 2: the minerals were the same as in example 15, but without the addition of microcapsules, i.e. only minerals and no microcapsules, a solution of 0.2g/mL was prepared with physiological saline (i.e. the concentration of minerals was the same as in group c).

f. Example 11 drug group: the composition of example 11 was mixed with physiological saline to make a 2.0g/mL suspension (i.e., NR at 0.48g/mL, NMN at 0.016g/mL, and minerals at 0.4 g/mL);

g. comparative group 3: microcapsules the same as in example 11, but without the addition of minerals, i.e. microcapsules only and no minerals, were made up in a 1.6g/mL suspension with physiological saline (i.e. NR and NMN concentrations the same as in group f);

h. comparative group 4: the minerals were the same as in example 11, but without the addition of microcapsules, i.e. only minerals and no microcapsules, a solution of 0.4g/mL was prepared with physiological saline (i.e. the concentration of minerals was the same as in group f).

i. Example 12 drug group: the composition of example 12 was mixed with physiological saline to prepare a suspension having a concentration of 2.0g/mL (i.e., NR concentration of 0.014g/mL, NMN concentration of 0.42g/mL, mineral concentration of 0.6 g/mL);

j. comparative group 5: microcapsules the same as in example 12, but without the addition of minerals, i.e. microcapsules only and no minerals, were made up in a 1.4g/mL suspension with physiological saline (i.e. NR and NMN concentrations the same as in group i);

k. comparative group 6: the minerals were the same as in example 12, but without the addition of microcapsules, i.e. only minerals and no microcapsules, a solution of 0.6g/mL was prepared with physiological saline (i.e. the concentration of minerals was the same as in group i).

Example 7 drug group: the composition of example 7 was mixed with physiological saline to make a 2.0g/mL suspension (i.e., NR at 0.10g/mL, NMN at 0.10g/mL, minerals at 0.0002 g/mL);

example 8 drug group: the composition of example 8 was mixed with physiological saline to make a 2.0g/mL suspension (i.e., NR at 0.40g/mL, NMN at 0.10g/mL, minerals at 0.002 g/mL);

example 9 drug group: the composition of example 9 was mixed with physiological saline to make a 2.0g/mL suspension (i.e., NR at 0.30g/mL, NMN at 0.30g/mL, minerals at 0.02 g/mL);

(2) results of the experiment

As shown in tables 4 and 5.

TABLE 4

TABLE 5

The results show that the MDA content and the serum SOD activity of the drug group have no statistical difference with those of a blank control group, which indicates that the drug combination can effectively enhance the capability of an organism for resisting oxidation and removing free radicals, reduce the degree of tissue cell damage, and basically recover the index to the level without aging and molding.

The research also finds that the MDA content of the comparison group is higher than that of the medicine group (P <0.05), the SOD activity is lower than that of the medicine group (P <0.05), the anti-oxidation and anti-aging effects are not better than that of the medicine group, the combination mode of the invention is optimal, and the lack of one component can reduce the anti-oxidation and anti-aging effects.

As can be seen from table 4, the percentage of MDA reduction in serum, liver and brain was 40.5%, 43.2% and 33.5% for comparative group 1 using the microcapsules containing NR and NMN alone, respectively; comparative group 2, using minerals alone, reduced the percentage of MDA in serum, liver and brain by 5.4%, 8.9% and 10.1%, respectively. The data for the drug group of example 15 of the present invention, however, show that when the microcapsules containing NR and NMN were used in combination with minerals, the percentage reduction of MDA in serum, liver and brain was 56.6%, 61.4% and 66.8%, respectively, which were significantly higher than the sum of the simple additive effects of reducing MDA of comparative group 1 and comparative group 2, 45.9% (40.5% + 5.4%), 52.1% (43.2% + 8.9%) and 43.6% (33.5% + 10.1%). The results of the comparative tests of the drug group of example 11 of the present invention and comparative groups 3 and 4, and the drug group of example 12 of the present invention and comparative groups 5 and 6 were similar. It can be seen that the pharmaceutical combination of the present invention has a significant effect on reducing MDA and a synergistic effect with respect to (NR + NMN) alone or mineral alone in combination.

As can be seen from table 5, the percentage of increase in SOD in serum, liver and brain was 72.6%, 44.1% and 33.6% for comparative group 1 using the microcapsules containing NR and NMN alone, respectively; comparative group 2, using minerals alone, increased the percentages of SOD in serum, liver and brain by 8.8%, 5.6% and 6.2%, respectively. The data of the drug group of the present invention, however, show that when the microcapsules containing NR and NMN were used in combination with minerals, the percentages of increased SOD in serum, liver and brain were 96.5%, 65.3% and 48.0%, respectively, which were significantly higher than the sum of simple additions of SOD increasing effects of comparative group 1 and comparative group 2 of 81.4% (72.6% + 8.8%), 49.7% (44.1% + 5.6%) and 39.8% (33.6% + 6.2%). The results of the comparative tests of the drug group of example 11 of the present invention and comparative groups 3 and 4, and the drug group of example 12 of the present invention and comparative groups 5 and 6 were similar. It can be seen that the pharmaceutical composition of the present invention has significant effect in increasing SOD, and has synergistic effect compared to (NR + NMN) alone or mineral alone, or the combination of (NR + NMN) and mineral.

Although specific embodiments of the invention have been described in detail, those skilled in the art will appreciate. Various modifications and substitutions of those details may be made in light of the overall teachings of the disclosure, and such changes are intended to be within the scope of the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.