Anti-aging NMN derivative as well as preparation process and application thereof

Abstract

The invention discloses an anti-aging NMN derivative. The anti-aging NMN derivative is a linoleic acid diester of NMN formed by esterification reaction of two alcoholic hydroxyl groups in an NMN molecule and linoleic acid. NMN and linoleic acid are esterified to obtain a new compound, the compound can exert antioxidant, anti-aging and other physiological activities on multiple mechanisms and has a synergistic effect, and in addition, due to introduction of non-polar linoleic acid, the skin permeation effect of NMN is improved, and the skin regeneration effect of NMN is improved. The water-soluble component nicotinamide mononucleotide is efficiently delivered into the skin to achieve the anti-aging effect. In addition, the invention also provides a preparation method of the NMN derivative and application of the NMN derivative in preparation of anti-aging drugs, anti-aging or anti-wrinkle cosmetics and anti-aging health-care products.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to an anti-aging NMN derivative and a preparation process and application thereof. Background Art

[0002] NMN (β-nicotinamide mononucleotide) is a naturally occurring bioactive nucleotide and a precursor of the longevity protein cofactor nicotinamide adenine dinucleotide (NAD+). NAD+ is involved in a variety of redox reactions in the human body and is a key molecule in cellular energy metabolism. Many aging-related diseases, such as cardiac ischemia-reperfusion, Alzheimer's disease, Parkinson's disease, type 2 diabetes, acute kidney injury, etc., are related to the reduction of NAD+ levels in the body. However, with age, the level of NAD+ in the body will gradually decrease, not only will cell function decline, but the skin will also age. Existing studies have shown that NMN can delay the aging of the human body, especially in skin tissue. NMN can slow down and promote keratinocyte metabolism to a certain extent, accelerate the decomposition of melanin, and make the skin whiter and more translucent.

[0003] However, NMN itself has a poor absorption rate in the skin surface, and it is difficult to obtain satisfactory results by direct application. In addition, NMN has poor stability and is easily decomposed or oxidized in water or air, resulting in reduced activity or failure. Most of the prior art uses a method of compounding NMN with other components to achieve the effect of improving the skin penetration efficiency and depth of NMN. However, this method generally has technical problems such as complex formula, high cost and limited effect. Summary of the invention

[0004] In order to solve the problems of poor absorption rate of NMN in the skin surface, poor stability, and easy decomposition or oxidation in the prior art, the present invention provides an anti-aging NMN derivative. NMN is esterified with linoleic acid to obtain a new compound, which can exert physiological activities such as antioxidant and anti-aging in multiple mechanisms and has a synergistic effect. In addition, due to the introduction of non-polar linoleic acid, the skin penetration effect of NMN is improved, and the water-soluble component nicotinamide mononucleotide is efficiently delivered into the skin to exert anti-aging effects. In addition, the present invention also provides a method for preparing the above-mentioned anti-aging NMN derivative and uses it to prepare anti-aging drugs, anti-aging or anti-wrinkle cosmetics, and anti-aging health products.

[0005] In order to achieve the above object, the present invention adopts the following technical solutions: In a first aspect of the present invention, an anti-aging NMN derivative is provided, wherein the anti-aging NMN derivative is a linoleic acid diester formed by esterification reaction between two alcoholic hydroxyl groups in the NMN molecule and linoleic acid respectively; Among them, the structural formulas of NMN and linoleic acid are as follows Figure 1 As shown; The structural formula of the linoleic acid diester of NMN is as follows Figure 2 shown.

[0006] Using the above technical solution: Linoleic acid (CAS: 60-33-3) is an ω-6 unsaturated fatty acid and one of the essential fatty acids for the human body. Linoleic acid has low polarity and strong lipophilicity. It can quickly penetrate the skin, help the skin firmly lock in moisture and nutrients, and prevent moisture loss on the skin surface causing dry skin. Studies have shown that linoleic acid can also effectively fight free radicals in the skin, reduce oxidative stress, and make the skin look younger and more radiant. In the human body, linoleic acid can be converted into prostaglandin E and leukotriene B, both of which can effectively inhibit inflammatory responses and help the body resist various inflammatory invasions.

[0007] In the present application, NMN and linoleic acid molecules are creatively linked by covalent bonds. On the one hand, they can exert their antioxidant, anti-aging and other physiological activities on the skin through multiple mechanisms, thereby having a synergistic effect. On the other hand, due to the introduction of the non-polar molecule linoleic acid, the skin penetration effect of NMN can be greatly improved, overcoming the defect of low skin penetration caused by the high water solubility of NMN, and achieving the efficient delivery of the water-soluble component nicotinamide mononucleotide into the skin to exert its anti-aging effect, thereby achieving the effects of improving skin aging, anti-oxidation and anti-aging.

[0008] The second aspect of the present invention provides a process for preparing the above-mentioned anti-aging NMN derivative, comprising the following steps: S1, linoleic acid reacts with N-hydroxysuccinimide to form an active ester; S2. Esterify the active ester generated in step S1 with the two alcoholic hydroxyl groups of NMN to obtain a linoleic acid diester of NMN.

[0009] Furthermore, the anti-aging NMN derivative and / or its salt is used to prepare a preparation with anti-aging effect.

[0010] Furthermore, the salt of the anti-aging NMN derivative includes one or a mixture of several of sodium salt, calcium salt, magnesium salt, potassium salt, iron salt, zinc salt or manganese salt.

[0011] Furthermore, the preparation includes medicines, cosmetics and / or health products.

[0012] Furthermore, the preparation includes at least one of an injection, a lyophilized powder injection, a capsule, a tablet, a granule, an ointment, a spray, and an oral preparation.

[0013] The third aspect of the present invention provides an anti-aging drug, the active ingredient of which includes the above-mentioned anti-aging NMN derivative and / or its salt.

[0014] In a fourth aspect of the present invention, an anti-aging or anti-wrinkle cosmetic is provided, wherein the anti-aging or anti-wrinkle cosmetic comprises the above-mentioned anti-aging NMN derivative and / or its salt.

[0015] The fifth aspect of the present invention provides an anti-aging health product, which includes the above-mentioned anti-aging NMN derivative and / or its salt.

[0016] Compared with the prior art, the present invention has the following beneficial effects: (1) The present invention creatively connects the two alcoholic hydroxyl groups in NMN with two linoleic acid molecules through ester bonds to obtain a new compound - linoleic acid diester of NMN, which can exert antioxidant, anti-aging and other physiological activities in multiple mechanisms and has a synergistic effect; (2) Due to the introduction of non-polar linoleic acid in the anti-aging NMN derivative, the skin penetration effect of NMN can be greatly improved, overcoming the defect of low skin penetration caused by the high water solubility of NMN, and achieving the efficient delivery of the water-soluble component nicotinamide mononucleotide into the skin to exert its anti-aging effect, thereby achieving the effects of improving skin aging, anti-oxidation and anti-aging. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

[0018] Figure 1 is the structural formula of NMN and linoleic acid in the present invention.

[0019] Figure 2 It is the structural formula of the linoleic acid diester of NMN in the present invention. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Example 1 The specific preparation process of the anti-aging NMN derivative (linoleic acid diester of NMN) is as follows: S1. Weigh 5.6 g (0.01 mol) of linoleic acid, dissolve it in 60 mL of THF (tetrahydrofuran), add 1.38 g (0.012 mol) of N-hydroxysuccinimide (HOSu), add 2.48 g (0.012 mol) of dicyclohexylcarbodiimide (DCC) in an ice-water bath, react at room temperature for 8 hours, after the reaction is completed, evaporate THF under reduced pressure, dissolve the reactant in 100 mL of dichloromethane, wash the dissolved solution with 100 mL of saturated sodium bicarbonate twice, continue to wash with 100 mL of saturated brine twice, and dry the organic phase with anhydrous sodium carbonate; S2. To the obtained dichloromethane solution, 2.24 g (0.002 mol) of 4-dimethylaminopyridine (DMAP) and 1.94 g (0.015 mol) of N,N-diisopropylethylamine (DIEA) were added in sequence, and then 3.34 g (0.01 mol) of NMN were added in batches. The reaction was allowed to react for 12 hours, and the reaction liquid was spin-dried. The reactant was purified by column chromatography (the eluent was a petroleum ether-ethyl acetate system) to obtain 4.60 g of a white solid, which was the linoleic acid diester of the target product NMN. The yield was calculated to be 53.6%.

[0022] Among them, the structural formulas of the reaction raw materials NMN and linoleic acid are as follows Figure 1 shown.

[0023] The target product obtained in step S2 was subjected to mass spectrometry and nuclear magnetic resonance analysis. The peak shown by mass spectrometry was 857.3 [M+ H]+, and the results of nuclear magnetic resonance analysis were as follows: 1H-NMR (400 MHz, DMSO) δ 11.87 (m, 1H), 9.52 (m, 1H), 9.28 (d, J = 6.2 Hz, 1H), 8.99 (m, 1H), 8.35 (m, 1H), 7.76 (m,2H), 6.21 (d, J = 5.5 Hz, 1H), 5.59-5.23(m, 8H), 4.63 (m, 1H), 4.59 (m, 1H),4.46 (m, 1H), 4.23 (m, 1H), 4.03 (m, 1H), 2.85 (m, 4H), 2.12 (t, J = 7.5 Hz,4H), 2.11-2.03 (m, 8H), 1.69-1.55 (m, 4H), 1.45-1.22 (m, 28H), 0.92-0.83 (m,6H).

[0024] The above mass spectrometry and nuclear magnetic resonance analysis results show that the linoleic acid diester of NMN has been successfully prepared, and its structural formula is as follows: Figure 2 shown.

[0025] Example 2 Antioxidant activity test The invasion of reactive oxygen is one of the main causes of skin aging. Reactive oxygen removes foreign matter by decomposing it in the process of removing foreign matter that enters the body. However, if too much reactive oxygen is produced due to various reasons, it will not only damage its own cell tissues, but also combine with unsaturated fatty acids in the body to form lipid peroxides. It can also combine with proteins and lipids in the skin to generate oxides, which leads to skin aging, and has a strong effect on the oxidation process of melanin, causing pigmentation. This example respectively detects the scavenging effects of the linoleic acid diester (NMNL) of the product NMN in Example 1, NMN and linoleic acid on DPPH free radicals (1,1-diphenyl-2-trinitrophenylhydrazine).

[0026] Test samples: NMNL, NMN and linoleic acid were prepared into 10% test solutions using ethanol-water (5:5), respectively.

[0027] The DPPH free radical scavenging experiment process is as follows: take 3mL of 0.2mmol / L DPPH-anhydrous ethanol solution in a cuvette, then add 1mL of the test sample, mix well, and keep it away from light at room temperature for 30min. Measure the absorbance at a wavelength of 517nm, and the absorbance value is recorded as A1. The absorbance value of only adding 3mL of anhydrous ethanol solution and 1mL of the test sample solution to the cuvette is recorded as A2, and the absorbance value of adding 3mL of DPPH-anhydrous ethanol solution and 1mL of anhydrous ethanol solution to the cuvette is recorded as A3.

[0028] Calculate the DPPH free radical scavenging rate according to the following formula: DPPH free radical scavenging rate (%) = [1-(A1-A2) / A3] × 100%, taking the average of three measurements.

[0029] The test results are shown in Table 1.

[0030] Table 1 DPPH free radical scavenging rate of NMNL, NMN and linoleic acid

[0031] It can be seen from the test results in Table 1 that the DPPH free radical scavenging rate of the linoleic acid diester of NMN (NMNL) in the present invention is significantly improved compared with both NMN and linoleic acid. NMNL can effectively scavenge free radicals and has antioxidant and anti-aging effects.

[0032] Example 3 Transdermal absorption rate test The transdermal absorption rate was determined using a Franz transdermal diffusion cell. Commercially available fragrant pig skin was fixed between the upper and lower chambers of the Franz transdermal diffusion cell, with the stratum corneum facing upward. NMNL and NMN were mixed with the cosmetic matrix and coated on a medical tape to prepare the test samples (drug loading of 200 μg / cm 2 ).

[0033] The test was conducted using the RT800 automatic sampling transdermal diffusion system. Physiological saline was placed in the receiving pool, and the receiving solution was just in contact with the dermis. Then 100 μL was taken at 3h, 6h, 9h and 12h, and the concentrations of NMNL and NMN in the receiving solution were detected by HPLC (high performance liquid chromatography), and the transdermal absorption amount was calculated. The transdermal absorption rate was calculated according to the following formula: Transdermal absorption rate (%) = (transdermal absorption amount / initial dose) × 100%, taking the average value of three measurements.

[0034] The test results are shown in Table 2.

[0035] Table 2 Transdermal absorption rate of NMNL and NMN

[0036] From the test results in Table 2, it can be seen that the unmodified NMN is difficult to pass through the skin due to its high polarity, and the permeability within 12 hours does not exceed 10%. The transdermal absorption rate of the modified compound NMNL is significantly improved, and the transdermal absorption rate is the highest at 12 hours (75.1%). Therefore, the linoleic acid diester of NMN in this application has a good skin permeability.

[0037] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An anti-aging NMN derivative, characterized in that: The anti-aging NMN derivative is a linoleic acid diester formed by esterification reaction between two alcoholic hydroxyl groups in the NMN molecule and linoleic acid respectively; The structural formula of the linoleic acid diester of NMN is as follows: 。 2. A process for preparing the anti-aging NMN derivative according to claim 1, characterized in that: The steps include: S1, linoleic acid reacts with N-hydroxysuccinimide to form an active ester; S2. Esterify the active ester generated in step S1 with the two alcoholic hydroxyl groups of NMN to obtain a linoleic acid diester of NMN.

3. A use of the anti-aging NMN derivative as claimed in claim 1, characterized in that: The anti-aging NMN derivative and / or its salt is used to prepare a preparation with anti-aging effect.

4. The use of the anti-aging NMN derivative according to claim 3, characterized in that: The salt of the anti-aging NMN derivative includes one or a mixture of several of sodium salt, calcium salt, magnesium salt, potassium salt, iron salt, zinc salt or manganese salt.

5. The use of the anti-aging NMN derivative according to claim 3, characterized in that: The preparations include medicines, cosmetics and / or health products.

6. The use of the anti-aging NMN derivative according to any one of claims 3 to 5, characterized in that: The preparation comprises at least one of an injection, a lyophilized powder injection, a capsule, a tablet, a granule, an ointment, a spray, and an oral preparation.

7. An anti-aging drug, characterized in that: The active ingredient of the anti-aging drug includes the anti-aging NMN derivative and / or its salt as described in claim 1.

8. An anti-aging or anti-wrinkle cosmetic, characterized in that: The anti-aging or anti-wrinkle cosmetic comprises the anti-aging NMN derivative and / or its salt as claimed in claim 1.

9. An anti-aging health product, characterized in that: The anti-aging health product includes the anti-aging NMN derivative and / or its salt as described in claim 1.

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