CN112679443A - Preparation method and application of fructosazine - Google Patents

Abstract

The invention discloses a preparation method and application of fructosazine, wherein the preparation method of the fructosazine comprises the steps of synthesis of 1-dibenzylamino-1-deoxy-D-fructose, preparation of 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose, preparation of isopropylidene protected fructosazine and synthesis of the fructosazine. The preparation method of the fructosazine provided by the invention uses Pd/C to catalyze amino sugar to oxidize to form pyrazine ring for the first time. The reaction route has the advantages of economic and easily-obtained raw materials, mild reaction conditions, avoidance of high temperature and high pressure, single product, high purity and avoidance of a complex post-treatment process. Furthermore, the invention provides the application of the fructosazine in preparing the anti-aging cosmetics.

Description

Preparation method and application of fructosazine

Technical Field

The invention belongs to the field of chemical preparations, and particularly relates to a preparation method and application of fructosazine.

Background

Fructosazine (FZ) and Deoxyfructosazine (DOF) are a class of pyrazine compounds that are found, isolated and characterized in baked foods. The compounds are useful as flavoring agents in the food industry due to their uniquely attractive odor; to increase or modify the taste of tobacco smoke due to its low odor threshold. Both of them have antibacterial activity in terms of pharmacological and physiological activities, and FZ plays an important role as a heat-resistant antibacterial agent; both also have inhibitory activity against interleukins (1 β and 2), which may be helpful in preventing pathological cartilage degradation and other inflammatory diseases; also, recent studies have shown that FZ has α -glucosidase inhibitory activity, and that FZ and DOF are responsible for transport by hexose transporters in human intestinal endothelial cells.

Due to their potential utility values, researchers have developed several synthetic methods associated with maillard reactions to prepare such pyrazine compounds. Typically formed by reacting a sugar or amino sugar with ammonia or an ammonium salt in an aqueous solution at a neutral or mildly acidic pH. Fujii et al in hot sodium methoxide at 65 deg.C, 2-amino-2-deoxy-D-glucose (GlcNH2) can be condensed to give FZ, crystallized overnight, in 9.6% yield after resin treatment. Sumoto et al will GlcNH₂The hydrochloride was dissolved in 28% aqueous ammonia and stirred at room temperature for 3 weeks to give a mixture of FZ and DOF with a FZ yield of 47%. GlcNH by Rohovec et al₂The hydrochloride reacts under the catalysis of phenylboronic acid or boric acid ester, and the mixture of FZ and DOF is obtained after the reaction in aqueous solution for 3 hours at room temperature, and the yield of DOF can reach 96%. L. et al used a basic ionic liquid, 1-butyl-3-methylimidazolium hydroxide ([ BMIM)]OH) as solvent and catalyst, a mixture of the two was synthesized with a mixed yield of 49%. In addition, y.hrynets et al use metal cation catalysis to acidify amino sugarsSelf-condensation in aqueous solution to give a mixture of FZ and DOF. Houqilin et al used 1-ethyl-3-methylimidazolium acetate ionic liquid catalyst, aqueous hydrogen peroxide as oxidant, GlcNH₂Dehydrogenated to obtain fructosazine, and reacted at 20 deg.c for 10 days in 40% yield. Wanmeifeng et al GlcNH under 4-pyridineboronic acid catalysis₂DOF can be selectively obtained after 6 hours of reaction at 60 ℃, and the yield is 89%.

As described above, the conventional reaction has a problem that a long reaction time or a high reaction temperature is required, and FZ cannot be selectively produced under most conditions. The reaction product is greatly influenced by the reaction conditions, particularly the reaction solvent, the pH value of the reaction mixture and the reaction temperature; and the new catalysts are difficult to prepare or expensive and are not economically available. In addition, both FZ and DOF are water-soluble substances, and thus are difficult to isolate and purify. Therefore, there is a need to develop a new synthetic method to prepare fructosazine with high selectivity and high yield. The invention selectively prepares a single product FZ by four-step reaction from simple and easily obtained monosaccharide raw material D-glucose.

Aging refers to the deterioration of tissue physiology and function over time, which is a major risk factor for human diseases such as cancer, diabetes, cardiovascular diseases and neurodegenerative diseases^[19]. According to the invention, the characterization of the aging marker of the human embryo lung diploid fibroblast proves that the fructosazine has certain anti-aging activity, and a basis is provided for subsequent related evaluation.

Disclosure of Invention

The invention aims to provide a preparation method and application of fructosazine, so as to solve the problems in the prior art.

The technical scheme of the invention is as follows:

in a first aspect, the invention provides a preparation method of fructosazine, which comprises the following steps:

step 1, synthesis of 1-dibenzylamino-1-deoxy-D-fructose

Adding D-glucose into absolute ethyl alcohol, stirring at room temperature, sequentially adding dibenzylamine and glacial acetic acid, uniformly stirring, refluxing for 3 hours at 80 ℃, cooling to room temperature, performing suction filtration, washing a filter cake for three times by using the absolute ethyl alcohol, and performing suction filtration to obtain (3S, 4R, 5R) -2- ((dibenzylamino) methyl) tetrahydro-2H-pyran-2, 3,4, 5-tetraol, namely 1-dibenzylamino-1-deoxy-D-fructose.

Step 2, preparation of 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose

Adding the 1-dibenzylamino-1-deoxy-D-fructose obtained in the step 1) and trimethyl orthoformate into acetone, and adding concentrated hydrochloric acid at 0 ℃. After stirring at 0 ℃ for 2 hours, the reaction mixture was neutralized with ammonia to neutrality, acetone was removed by rotary evaporation, extracted with dichloromethane, and the organic phase was dried to give 1-dibenzylamino-1-deoxy-4, 5-oxo-isopropylidene-D-fructose as a colorless oil.

Step 3, preparation of isopropylidene protected fructosazine

Dissolving the 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose obtained in the step 2) in absolute ethyl alcohol by using N₂Replacing three times, adding 5-30 percent by mass of Pd/C (5-15 percent by mass of Pd content) and H of 1-dibenzylamino-1-deoxy-4, 5-oxygen-isopropylidene-D-fructose₂Displacement is carried out for three times; at 10-80 ℃ in H₂Stirring for 8-16 hours under the condition; thin layer chromatography monitoring disappearance of starting material and H removal₂Introducing air into the reaction environment, and continuously stirring for 8-16 hours; and (3) filtering to remove Pd/C, performing rotary evaporation to remove ethanol, washing the obtained crude product by dichloromethane, and filtering to obtain a light yellow solid, namely isopropylidene protected fructosazine, and pyrazine-2, 5-bis- ((1R, 2S, 3R) -2, 3-oxygen-isopropylidene-1, 2,3, 4-butanetetraol).

Step 4, synthesis of fructosazine

Taking the isopropylidene protected fructosazine obtained in the step 3), adding 80% acetic acid solution, wherein each 100mg of reactant corresponds to 10-100mL of solvent, stirring at 10-80 ℃ for 6-10 hours, removing the solvent by rotary evaporation to obtain white solid, carrying out suction filtration, washing with methanol, and drying to obtain (1R, 1'R, 2S, 2' S, 3R, 3'R) -1,1' - (2, 5-pyrazine) -bis-1, 2,3, 4-butanetetraol, namely the fructosazine.

In a second aspect, the present invention provides the use of a fructosazine in the preparation of a cosmetic for anti-aging.

In a third aspect, the present invention provides an anti-aging cosmetic formulation comprising a fructosazine and a cosmetically acceptable carrier.

In some specific embodiments, the anti-aging is anti-skin aging.

The anti-aging cosmetic formulation may be in any convenient form without particular limitation. In a preferred embodiment, the cosmetic formulation is in the form of a care or cosmetic in the form of a solution, lotion, cream, gel, ointment, foam, soap, eye shadow, lipstick, shower gel, shaving cream, etc. for topical application, which may be applied via application to the surface of the skin. In a preferred embodiment, the cosmetic formulation is in the form of an infusion solution for injection subcutaneously or intravenously, such as a suspension, an intravenous infusion solution, a dry powder injection, a suspension injection, a dry powder suspension injection and the like.

By "cosmetically acceptable carrier" is meant an inert substance used as a carrier for the fructosazine, which is not toxic, irritating, pyrogenic, antigenic and hemolytic to the site of application (e.g., skin), and which has no substantial pharmacological activity and does not interfere with the beneficial effects of the fructosazine. Generally, the cosmetically acceptable carrier is used in an amount of about 1% to about 99.9%, preferably about 50% to about 99%, based on the total weight of the cosmetic formulation. Suitable types of cosmetically acceptable carriers depend on the form of the cosmetic formulation.

In a fourth aspect, the present invention provides the use of a fructosazine in the manufacture of a medicament for anti-aging.

The invention has the advantages that:

in view of the fact that fructosazine finds various pharmacological activities at present, starting from common monosaccharide D-glucose, through four-step reaction of Amadori rearrangement, acetone isopropylidene protection and Pd/C catalytic oxidation ring closure, isopropylidene protection is removed, so that a single product fructosazine is successfully obtained, and the total yield is 16%. Wherein the key step is to use Pd/C to catalyze amino sugar to be oxidized to form pyrazine ring for the first time. The key intermediate of the tetrafosmin has been sufficiently researched in the structural analysis. The reaction route has the advantages of economic and easily-obtained raw materials, mild reaction conditions, avoidance of high temperature and high pressure, single product, high purity and avoidance of a complex post-treatment process. By measuring the aging marker beta-galactosidase, the fructosazine shows good anti-aging activity at the early stage or the middle stage of cell aging under the condition of 1 mu M, and provides a foundation for the subsequent research on the anti-aging activity of the compounds.

Drawings

Fig. 1 is a synthesis scheme of the preparation method of fructosazine provided in the examples of the present invention.

Fig. 2 is a schematic crystal structure diagram of a compound iv with an atom number in the preparation method of fructosazine provided in the embodiment of the present invention.

Fig. 3 is a schematic diagram showing the effect of different concentrations of FZ on 2BS cell viability using MTT method, where n is 3, x ± s; p <0.01 compared to control.

FIGS. 4A and 4B are photographs of SA- β -Gal staining of cells in which PD38 started to be cultured with drugs; wherein, FIG. 4A is a control group; FIG. 4B is the 1 μ M FZ group; cells were photographed under a 10 × 10 microscope.

Fig. 4C is a statistical image of SA- β -Gal staining of cells in PD38 primed culture, P < 0.01.

FIGS. 5A and 5B are photographs of SA- β -Gal staining of PD46 cells: FIG. 5A is a control group; FIG. 5B is the 1 μ M FZ group; cells were photographed under a 10 × 10 microscope.

Fig. 5C is a statistical representation of the SA- β -Gal staining of PD46 cells, P < 0.01.

Detailed Description

The invention is further illustrated by the following examples, but not by way of limitation, in connection with the accompanying drawings. The following provides specific materials and sources thereof used in embodiments of the present invention. However, it should be understood that these are exemplary only and not intended to limit the invention, and that materials of the same or similar type, quality, nature or function as the following reagents and instruments may be used in the practice of the invention. The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

Reagent and apparatus

Reagents and instruments referred to in the following examples include, but are not limited to:

d-glucose, analytical grade, komi european chemical reagent (tianjin); absolute ethyl alcohol, methanol, acetone, hydrochloric acid, glacial acetic acid, dichloromethane, analytically pure, chemical industry of Fuyu (Tianjin); dibenzylamine, 10% Pd/C, analytical grade, annaiji chemical reagent (shanghai); nitrogen and hydrogen, high purity, and large-scale light-coupled special gas plants.

Vaian DLG400 nmr instrument, Varian corporation (usa); LC-MS 1260 and 6130, Agilent, USA; n-1001 rotary evaporator, Shanghai EYELA instruments Inc.; waters e2695 liquid chromatograph, 2489 ultraviolet detector; GM-1.0A diaphragm vacuum pump, available from Jinteng laboratory facilities, Inc., Tianjin; BS210S precision electronic balance, beijing sidoris balance ltd; ZF-TA Portable ultraviolet analyzer, Consumer City prosperous instrument.

Thiazole blue (MTT), bio-technical institute of cloudy sky; age-related beta-galactosidase (SA-beta-Gal) staining kit, Biyuntian biotechnological research institute.

Example 1: synthesis of fructosazine

Fig. 1 is a synthesis scheme of the preparation method of fructosazine provided in the examples of the present invention, see fig. 1.

In this example, the synthesis of fructosazine comprises the following steps:

step 1, synthesis of 1-dibenzylamino-1-deoxy-D-fructose^[20]

Weighing 3.6g D-glucose (compound I) (D-glucose,20mmol), adding into a three-neck flask, adding 40mL of absolute ethyl alcohol, stirring at room temperature, sequentially adding 3.96g of dibenzylamine and 1mL of glacial acetic acid, stirring uniformly, refluxing at 80 ℃ for 3 hours, cooling to room temperature, carrying out vacuum filtration, washing a filter cake with 3 x 10mL of absolute ethyl alcohol, and carrying out vacuum filtration to obtain (3S, 4R, 5R) -2- ((di-n-ethyl acetate)Benzylamino) methyl) tetrahydro-2H-pyran-2, 3,4, 5-tetraol 5.9g, i.e. 1-dibenzylamino-1-deoxy-D-fructose (compound II), yield 82%.¹H NMR(400 MHz,DMSO-d6)δ7.30(m,10H,Ph-H),5.25(s,1H,OH),4.43(d,J＝4.7Hz,1H, H-4),4.38(s,1H,OH),4.15(t,J＝4.7Hz,1H,H-5),3.81(s,1H,OH),3.74(d,J＝13.8Hz,2H,H-14),3.63(d,J＝13.8Hz,2H,H-7),3.58(d,2H,H-1),3.43(d,1H, H-3),2.68(q,J＝13.3Hz,2H,H-6)。

Step 2, synthesis of 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose

Compound II (2.5g,6.96mmol) obtained in step 1) and trimethyl orthoformate (2.0mL,18.3 mmol) were added to acetone (80mL) and concentrated hydrochloric acid (0.6mL) was added at 0 ℃. After stirring at 0 ℃ for 2 hours, the reaction mixture was neutralized with aqueous ammonia, acetone was removed by rotary evaporation, 3X 50mL of dichloromethane was extracted, and the organic phase was dried to give 1.94g of 1-dibenzylamino-1-deoxy-4, 5-oxo-isopropylidene-D-fructose (compound III) as a colorless oil with a yield of 70%.¹H NMR(400MHz,DMSO-d6)δ7.33-7.25(m,10H, Ph-H),5.53(d,J＝1.1Hz,OH),4.78(d,J＝5.7Hz,1H,H-4),4.11(dd,J＝5.7,2.3 Hz,1H,H-5),4.03–3.93(m,2H,H-1),3.75(d,J＝14Hz,2H,H-10),3.75(1H, OH),3.60(d,J＝14Hz,2H,H-17),3.45(t,1H,H-3),2.69(d,J＝2.3Hz,2H,H-6,), 1.38(s,3H,H-8),1.26(s,3H,H-9)。

Step 3, preparation of isopropylidene protected fructosazine

In a 100mL round-bottom flask, the compound III (2.0g,5.01mmol) obtained in step 2) was weighed out and dissolved in absolute ethanol (30mL) using N₂Three times displacement, 10% Pd/C (0.2g), H₂The substitution was carried out three times. At room temperature, H₂Stirred under conditions for 12 hours. Thin Layer Chromatography (TLC) monitors disappearance of starting material and H is removed₂The reaction was vented to ambient and stirred at room temperature for an additional 12 hours. Vacuum filtering to remove Pd/C, rotary evaporating to remove ethanol, washing the obtained crude product with dichloromethane, and filtering to obtain 522mg of compound IV, light yellow solid, pyrazine-2, 5-bis- ((1R, 2S, 3R) -2, 3-oxo-isopropylidene-1, 2,3, 4-butanetetraol), namely isopropylidene protected fructosazine, with the yield of 56%.¹H NMR(400MHz,DMSO-d6)δ8.63(d,J＝4.4Hz,2H,H-3,6), 5.53(d,J＝13.4,6.0Hz,1’-OH),4.99(t,1’-H),4.81(m,7’,7”-OH),4.45(dd,J＝4.2, H-7’,7”),4.30–4.09(m,1H,H-7’,7”),3.91–3.60(m,4H,H-2’,2”,6’,6”),1.49– 1.37(m,6H,H-4’,4”),1.21(d,J＝25.5Hz,6H,H-4’,4”).¹³C NMR(100MHz, DMSO-d6)δ155.46(C2,5),141.76(C3,6),107.60(C4’,4”),78.85(C1’,1”),77.70 (C2’,2”),70.65(6’,6”),59.78(7’,7”),26.76(C4’,4”-C1),25.38(C4’,4”-C2). MS(API-ES):m/z[M+H]⁺401.1,m/z[M+Na]⁺423.1。

In the step, Pd/C is firstly subjected to catalytic hydrogenation reduction debenzylation in the presence of hydrogen to generate fructosamine; and then, removing hydrogen to enable the reaction to be carried out in the air, wherein Pd/C plays a role in catalytic oxidation, so that fructosamine generates dihydropyrazine through self-condensation at room temperature under the condition of ethanol, and then generates pyrazine ring through oxidation, thereby realizing the generation of key intermediate fork protection fructosazine under the mild condition.

Characterization of the Crystal Structure of Compound IV

The compound IV is subjected to single crystal culture (dichloromethane/diethyl ether) and subjected to X-ray diffraction to determine the structure (CCDC 2055966), and the single crystal diffraction result shows that the compound IV has a correct structure (figure 2, table 1). Therefore, the success of the amino sugar oxidative ring closure reaction under the catalysis of Pd/C which is a key reaction step can be judged by combining the mass spectrum data and nuclear magnetic information of the compound and the single crystal data of the compound.

TABLE 1 Crystal Structure data for Compound IV

Step 4, synthesis of fructosazine

Weighing the compound IV (100mg) obtained in the step 3) into a 100mL reaction bottle, adding 80% acetic acid solution (30mL), stirring for 8 hours at 30 ℃, rotary evaporating to remove the solvent to obtain a white solid, vacuum filtering, washing with 3 × 10mL of methanol, filtering, and drying to obtain 41mg of compound V, (1R, 1'R, 2S, 2' S, 3R, 3'R) -1,1' - (2, 5-pyrazine) -bis-1, 2,3, 4-butanetetraol, namely fructosazine, with the yield of 51%.¹H NMR (600MHz,DMSO-d6)δ8.62(s,2H,H-3,6),5.33(d,J＝6.3Hz,2H,1’,1”-OH),4.96 (d,J＝6.2Hz,2H,2’,2”-OH),4.66(d,J＝4.5Hz,2H,3’,3”-OH),4.43(d,J＝7.3Hz, 4’4”-OH),4.39(t,J＝5.2Hz,2H,H-1’,1”),3.67-3.60(m,2H,H-2’,2”),3.61-3.50 (m,4H,H-3’,3”,4’,4”),3.42(dt,J＝10.2,4.9Hz,H-4’,4”).MS(API-ES):m/z [M+H]⁺320.1,m/z[M+Na]⁺323.1。

Example 2: fructosazine anti-aging Activity evaluation

First, experiment method

1. Cell culture

The 2BS cell line isolated from human fetal lung fibroblasts was originally established by the chinese bioproduct institute and is widely used in cell senescence models. It is considered that the cells are young cells under PD (population doubling) 30 and the replicating senescent cells are PD55 or more. The experimental cells were purchased from China center for type culture Collection of Wuhan university, and grown in MEM supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin double antibody, and placed at 37 deg.C and 5% CO₂In an incubator. When the cells grew to 85%, the cultured cells were divided at a ratio of 1:2 or 1:4 for passaging. Cumulative cell doubling times (CPDs) were calculated as log₂(D/D₀) Wherein D and D₀Cell density at harvest and seeding, respectively.

2. MTT assay

2BS cells were arranged at 3X 10³The/well density was seeded in flat bottom 96-well plates. After 24 hours, the cells were placed in culture media containing different concentrations of the drug and cultured for 48 hours. mu.L of MTT at a concentration of 5mg/mL was added to each well. After incubation for 4 hours in the dark, the drug solution in the wells was aspirated, and 200. mu.L of DMSO was added to each well. Measuring OD value with microplate reader, detecting wavelength of 570nm, and detecting temperature of 37 deg.C.

3. Senescence-associated beta-galactosidase (SA-beta-gal) staining^[21]

The principle of the experimental method is based on the specific high-activity expression of SA-beta-Gal in senescent cells, and X-Gal in a staining agent is catalyzed by the SA-beta-Gal to generate a blue product which can be seen under a microscope, so that the senescent cells are stained, and the senescent cells, resting cells, immortal cells or tumor cells are not stained.

The method is operated according to the instruction of a 'cell senescence beta-galactosidase staining kit', and mainly comprises the following steps: 2BS cells were arranged at 2X 10⁴The drug is added into the six-hole plate after changing the liquid. The culture medium was aspirated, washed once with PBS, and then 1mL of beta-galactosidase staining fixative was added and fixed for 15 minutes at room temperature. The cell fixative was aspirated and the cells were washed 3 times with PBS for 3 minutes each. PBS was aspirated and 1mL of staining solution was added to each well. 37 ℃ incubator (CO free)₂) Incubate overnight and seal the plate with parafilm to prevent evaporation. Observed under a microscope and recorded by 100 times photography. The positive cells stained blue are the senescent cells.

The aging ratio (blue staining rate) was calculated according to the following formula: the aging ratio is the number of blue cells in the photograph/total number of cells in the photograph × 100.

Second, experimental results

1. Optimal concentration of FZ and its effect on cell growth and proliferation potential

FZ can up-regulate cell activity under 5 mu M, has maximum promotion effect under the concentration of 1 mu M, can up-regulate 25 percent, and has statistical significance (P < 0.05). There was a slight but insignificant inhibition of proliferation at 5-50 μ M. Therefore, 1. mu.M was selected as the optimum concentration for the long-term culture of cells.

2. Effect of FZ on SA-beta-gal senescence-associated markers

2BS cells of PD38 at 2X 10⁴Each well was inoculated into 6-well plates and cultured for 10 days continuously, MEM medium containing 1. mu.M FZ was added to each of the drug-changing groups, and MEM medium was added to the blank group for culture. The blue dyeing rate of the blank group is obviously higher, statistics show that the blue dyeing rate of the drug-adding group is 25%, and the blue dyeing rate of the blank group is 44% higher than that of the drug-adding group by 19%. Experimental results prove that FZ can effectively delay cell senescence in the cell senescence process.

The 2BS cells of PD32 were cultured in a divided manner up to 46 generations, and MEM medium containing 1. mu.M FZ was added to each of the drug-changing groups, and MEM medium was added to the blank group for culture. It can be seen that the blue staining rate of the blank group is significantly higher. Statistics show that the blue dyeing rate of the drug adding group is 8%, and the blank group is 23% higher than the drug adding group by 15%. The experimental results demonstrated that FZ can delay senescence early in cellular senescence (results see fig. 4A to 5C).

Meanwhile, the comparison of two groups of experimental results can prove that the FZ can effectively delay the senility in the early cell stage or the middle cell stage, and the anti-aging effect is more obvious when the medicine is added and cultured in the early aging stage.

The above description of exemplary embodiments has been presented only to illustrate the technical solution of the invention and is not intended to be exhaustive or to limit the invention to the precise form described. Obviously, many modifications and variations are possible in light of the above teaching to those skilled in the art. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to thereby enable others skilled in the art to understand, implement and utilize the invention in various exemplary embodiments and with various alternatives and modifications. It is intended that the scope of the invention be defined by the following claims and their equivalents.

Claims (10)

1. The preparation method of fructosazine is characterized by comprising the following steps:

step 1, synthesis of 1-dibenzylamino-1-deoxy-D-fructose

Adding D-glucose into absolute ethyl alcohol, stirring at room temperature, sequentially adding dibenzylamine and glacial acetic acid, uniformly stirring, refluxing for 3 hours at 80 ℃, cooling to room temperature, performing suction filtration, washing a filter cake for three times by using the absolute ethyl alcohol, and performing suction filtration to obtain (3S, 4R, 5R) -2- ((dibenzylamino) methyl) tetrahydro-2H-pyran-2, 3,4, 5-tetraol, namely 1-dibenzylamino-1-deoxy-D-fructose;

step 2, preparation of 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose

Adding the 1-dibenzylamino-1-deoxy-D-fructose obtained in the step 1) and trimethyl orthoformate into acetone, and adding concentrated hydrochloric acid at 0 ℃; stirring at 0 deg.C for 2 hr, neutralizing the reaction mixture with ammonia water to neutrality, rotary evaporating to remove acetone, extracting with dichloromethane, and drying organic phase to obtain colorless oily substance 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose;

step 3, preparation of isopropylidene protected fructosazine

Dissolving the 1-dibenzylamino-1-deoxy-4, 5-oxy-isopropylidene-D-fructose obtained in the step 2) in absolute ethyl alcohol by using N₂Replacing three times, adding 5-30 percent of Pd/C and H by mass percent of 1-dibenzylamino-1-deoxy-4, 5-oxygen-isopropylidene-D-fructose₂Displacement is carried out for three times; at 10-80 ℃ in H₂Stirring for 8-16 hours under the condition; thin layer chromatography monitoring disappearance of starting material and H removal₂Introducing air into the reaction environment, and continuously stirring for 8-16 hours; performing suction filtration to remove Pd/C, performing rotary evaporation to remove ethanol, washing the obtained crude product with dichloromethane, and filtering to obtain a light yellow solid, namely isopropylidene protected fructosazine, pyrazine-2, 5-bis- ((1R, 2S, 3R) -2, 3-oxygen-isopropylidene-1, 2,3, 4-butanetetraol);

step 4, synthesis of fructosazine

Taking the isopropylidene protected fructosazine obtained in the step 3), adding 80% acetic acid solution, wherein each 100mg of reactant corresponds to 10-100mL of solvent, stirring at 10-80 ℃ for 6-10 hours, removing the solvent by rotary evaporation to obtain white solid, performing suction filtration, washing with methanol, and drying to obtain (1R, 1'R, 2S, 2' S, 3R, 3'R) -1,1' - (2, 5-pyrazine) -bis-1, 2,3, 4-butanetetraol, namely the fructosazine.

2. The method of claim 1, wherein the Pd content in Pd/C is 5% -15%.

3. The process for producing fructosazine according to claim 1 or 2, wherein in step 3), 1-dibenzylamino-1-deoxy-4, 5-oxo-isopropylidene-D-fructose obtained in step 2) is dissolved in absolute ethanol using N₂Replacing three times, adding 10 percent Pd/C and H of 10 percent of 1-dibenzylamino-1-deoxy-4, 5-oxygen-isopropylidene-D-fructose in mass percent of 10 percent₂Displacement is carried out for three times; at room temperature, H₂Stirring for 12 hours under the condition; thin layer chromatography monitoring disappearance of starting material and H removal₂Introducing air into the reaction environment, and continuously stirring for 12 hours; and (3) filtering to remove Pd/C, performing rotary evaporation to remove ethanol, washing the obtained crude product by dichloromethane, and filtering to obtain light yellow solid pyrazine-2, 5-bis- ((1R, 2S, 3R) -2, 3-oxygen-isopropylidene-1, 2,3, 4-butanetetraol), namely isopropylidene protected fructosazine.

4. The process according to claim 1 or 2, wherein the isopropylidene protected fructosazine obtained in step 3) is taken, 80% acetic acid solution is added, 30mL of solvent is added per 100mg of reaction product, stirring is carried out at 30 ℃ for 8 hours, the solvent is removed by rotary evaporation to obtain white solid, methanol washing and drying are carried out to obtain (1R, 1'R, 2S, 2' S, 3R, 3'R) -1,1' - (2, 5-pyrazine) -bis-1, 2,3, 4-butanetetraol, i.e. fructosazine.

5. The method for preparing fructosazine according to claim 3, wherein the isopropylidene protected fructosazine obtained in step 3) is taken, 80% acetic acid solution is added, 30mL of solvent is added for each 100mg of reactant, stirring is carried out for 8 hours at 30 ℃, the solvent is removed by rotary evaporation to obtain white solid, vacuum filtration, methanol washing filtration and drying are carried out to obtain (1R, 1'R, 2S, 2' S, 3R, 3'R) -1,1' - (2, 5-pyrazine) -bis-1, 2,3, 4-butanetetraol, namely fructosazine.

6. Use of fructosazine for the preparation of anti-aging cosmetic.

7. The use according to claim 6, wherein the anti-ageing is anti-skin ageing.

8. Use according to claim 6 or 7, wherein the fructosazine is prepared by the process according to any one of claims 1 to 5.

9. An anti-aging cosmetic formulation comprising a fructosazine and a cosmetically acceptable carrier.

10. Use of fructosazine in the manufacture of a medicament for anti-aging.

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