Supramolecule constructed by quercetin and ergothioneine as well as preparation method and application of supramolecule

Abstract

The invention belongs to the technical field of supramolecules, and discloses a supramolecule constructed by quercetin and ergothioneine as well as a preparation method and application of the supramolecule. The preparation method of the supramolecule constructed by quercetin and ergothioneine comprises the following steps: respectively dissolving quercetin and ergothioneine in a solvent to obtain a quercetin solution and an ergothioneine solution; the preparation method comprises the following steps: adding an ergothioneine solution into a quercetin solution in an inert atmosphere, mixing, carrying out an ionizing salt formation reaction under a heating condition, concentrating, distilling, crystallizing, filtering, washing and drying to obtain the supramolecule constructed by quercetin and ergothioneine. According to the invention, quercetin is taken as an anion precursor, ergothioneine is taken as a cation precursor, the supramolecular quercetin-ergothioneine is synthesized through ionizing salt formation reaction, the supramolecular quercetin-ergothioneine has excellent biocompatibility, solubility, skin permeability and high bioavailability, a host molecular skeleton of a quercetin monomer is completely reserved, and the supramolecular quercetin-ergothioneine can be used for preparing the supramolecular quercetin-ergothioneine. Meanwhile, the quercetin and ergothioneine compound has biological activity and effects of quercetin and ergothioneine.

Description

Technical Field

[0001] The invention relates to the technical field of supramolecular materials, and in particular to a preparation method and application of supramolecular materials constructed by quercetin and ergothioneine. Background Art

[0002] In recent years, supramolecular technology has begun to be applied in the fields of cosmetics and medicine. Compared with traditional physical mixing or chemical contracting methods, the most prominent advantage of supramolecular technology is that the molecules are connected by forces, which makes them have double or more effects. As a new synthesis technology, supramolecular technology has the advantages of penetrating the stratum corneum, increasing solubility, improving stability and bioavailability, and avoiding the first-pass effect in the liver. It is an excellent way in the fields of cosmetics and medicine.

[0003] In supramolecular technology, the interaction between supramolecular molecules can not only effectively penetrate the stratum corneum, but also improve its bioavailability as an active ingredient. Compared with traditional drug delivery strategies (oral, injection), transdermal drug delivery has many advantages, such as avoiding first-pass liver clearance, improving patient dependence on drugs and reducing side effects. However, due to the insolubility or slight solubility of macromolecules or poorly soluble compounds in water or most organic solvents, transdermal delivery is difficult. It is well known that macromolecular drugs have never entered the clinical trial stage due to their difficulty in delivery and low bioavailability.

[0004] Therefore, the prior art still needs to be improved and developed. Summary of the invention

[0005] In view of the above-mentioned deficiencies in the prior art, the object of the present invention is to provide a supramolecule constructed of quercetin and ergothioneine and a preparation method and application thereof, aiming to solve the problems of difficulty in transdermal delivery and low bioavailability of macromolecular or poorly soluble compounds.

[0006] The technical solution of the present invention is as follows:

[0007] The first aspect of the present invention provides a method for preparing a supramolecule constructed of quercetin and ergothioneine, comprising the steps of:

[0008] dissolving quercetin in a first solvent to obtain a quercetin solution;

[0009] Dissolving thioneine in a second solvent to obtain an thioneine solution;

[0010] Under an inert atmosphere, the thioneine solution is added to the quercetin solution and mixed, and an ionized salt-forming reaction is carried out under heating conditions;

[0011] The solution after the reaction is concentrated, distilled and crystallized, and then filtered, washed and dried to obtain a supramolecular quercetin and ergothioneine, which can be recorded as supramolecular quercetin-ergothioneine.

[0012] Optionally, the molar ratio of the quercetin to the ergothioneine is 1:1-1:10.

[0013] Optionally, the first solvent is one or more of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution; the second solvent is one of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution.

[0014] Optionally, the concentration of the quercetin solution is 1-10 mol / L; the concentration of the ergothioneine is 1-10 mol / L.

[0015] Optionally, the inert atmosphere is nitrogen or argon.

[0016] Optionally, the temperature of the ionization salt-forming reaction is 30-100° C., and the time of the ionization salt-forming reaction is 4-24 h.

[0017] Optionally, the step of concentrating the solution after the reaction and performing reduced pressure distillation and crystallization comprises: rotary evaporating the solution after the reaction of quercetin and thioneine under vacuum, and then performing reduced pressure distillation and crystallization when the volume of the solution is concentrated to 1 / 10-1 / 5 of the original volume.

[0018] Optionally, the drying method is vacuum drying, the drying temperature is 30-100° C., and the drying time is 24-48 hours.

[0019] The second aspect of the present invention provides a supramolecule constructed by quercetin and ergothioneine, which is prepared by the preparation method of the supramolecule constructed by quercetin and ergothioneine.

[0020] The supermolecule constructed by quercetin and ergothioneine is assembled by combining quercetin molecules and ergothioneine molecules through non-covalent bonding.

[0021] The third aspect of the present invention provides an application of the supramolecule constructed by quercetin and ergothioneine in the preparation of cosmetics or drug carriers.

[0022] Beneficial effect: the present invention uses quercetin as an anion precursor and ergothioneine as a cationic precursor, and synthesizes the supramolecular structure of quercetin and ergothioneine by concentrated crystallization after ionization salt-forming reaction, i.e. supramolecular quercetin-ergothioneine. The supramolecular quercetin-ergothioneine has a supramolecular structure combined by non-covalent bond action, so that it significantly improves the biocompatibility, solubility, skin permeability and bioavailability of quercetin and ergothioneine. The supramolecular quercetin-ergothioneine not only completely retains the main molecular skeleton of quercetin monomer, but also has the biological activity and efficacy of quercetin and ergothioneine at the same time, and enhances its application effect, and has good potential application value in the field of transdermal administration and drug delivery. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 The present invention is a schematic flow diagram of a method for preparing a supramolecular (supramolecular quercetin-ergothioneine) constructed by quercetin and ergothioneine.

[0024] Figure 2 This is the H NMR spectrum of the supramolecular quercetin-ergothioneine (molar ratio 1:2) prepared in Example 1 of the present invention.

[0025] Figure 3 The two-dimensional H-NMR spectrum of the supramolecular quercetin-ergothioneine (molar ratio 1:2) prepared in Example 1 of the present invention.

[0026] Figure 4 This is the H-NMR spectrum of the supramolecular quercetin-ergothioneine (molar ratio 1:1) prepared in Example 2 of the present invention.

[0027] Figure 5 The two-dimensional H-NMR spectrum of the supramolecular quercetin-ergothioneine (molar ratio 1:1) prepared in Example 2 of the present invention.

[0028] Figure 6 This is a diagram of the transdermal efficiency of the supramolecular quercetin-ergothioneine prepared in Example 1 of the present invention.

[0029] Figure 7 This is a cell viability diagram of the supramolecular quercetin-ergothioneine prepared in Example 1 of the present invention.

[0030] Figure 8 This is the solubility diagram of the supramolecular quercetin-ergothioneine prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0031] The present invention provides a kind of supramolecule constructed by quercetin and ergothioneine and its preparation method and application, in order to make the purpose, technical scheme and effect of the present invention clearer and clearer, the present invention is further described in detail below.It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0032] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used in the specification of the present invention herein are only for the purpose of describing specific embodiments and are not intended to limit the present invention.

[0033] The embodiment of the present invention provides a method for preparing a supramolecule constructed by quercetin and ergothioneine, comprising the steps of:

[0034] S1, dissolving quercetin in a first solvent to obtain a quercetin solution;

[0035] S2, dissolving thioneine in the second solvent to obtain thioneine solution;

[0036] S3, under inert atmosphere, the thioneine solution is added to the quercetin solution and mixed, and ionization salt-forming reaction is carried out under heating condition;

[0037] S4, the solution after the reaction is concentrated, distilled and crystallized, and then filtered, washed and dried to obtain a supramolecular quercetin and thioneine, which is recorded as supramolecular quercetin-thioneine.

[0038] The quercetin, also known as quercetin, is a flavonol compound widely distributed in the plant kingdom and has multiple biological activities. Quercetin has antioxidant function, such as preventing lipid oxidation, reducing glutathione consumption, improving antioxidant enzyme activity, inhibiting lipid peroxidation to protect liver cells from alcoholic oxidative damage, and the protective effect is a dose-time effect relationship; anti-inflammatory effect, can reduce the body's immune response to allergens, inhibit IgE-mediated allergic mediators from mast cells and IgG-mediated histamine release; antibacterial effect, has a broad spectrum of antibacterial properties, and the antibacterial effect on Gram-negative bacteria is stronger than Gram-positive bacteria; skin color enhancement and skin protection, using it in cosmetics, can play a unique role in protecting the skin, can keep the skin moisturized, smooth, delicate, tender, elastic, and has anti-wrinkle, anti-wrinkle, beauty care and recovery of skin physiological functions. The ergothioneine is a natural antioxidant that can protect cells in the human body and is an important active substance in the body. Ergothioneine can also remove free radicals, detoxify, maintain the biosynthesis of DNA, the normal growth of cells and multiple physiological functions such as cellular immunity. Described ergothioneine is a natural antioxidant, which also has good anti-oxidation and repair protection effects on skin. However, the solubility and transdermal effect of single quercetin are poor, thereby limiting its application effect and application range.

[0039] The present invention uses quercetin as an anion precursor and ergothioneine as a cationic precursor, synthesizes supramolecular quercetin-ergothioneine through ionization salt-forming reaction, and realizes separation and purification of products through concentration-recrystallization after the reaction is completed, so as to obtain a pure natural supramolecular with excellent antioxidant, soothing, repairing, anti-wrinkle and other properties. The supramolecular quercetin-ergothioneine has good biocompatibility, good solubility, high bioavailability and skin permeability; and the supramolecular quercetin-ergothioneine completely retains the main molecular skeleton of the quercetin monomer, while retaining the biological activity and efficacy of two substances of quercetin and ergothioneine, and effectively improves the solubility and transdermal degree of quercetin and ergothioneine, and greatly improves the biological activity, enhances its application effect, and has potential application value in transdermal administration and transportation.

[0040] In some embodiments, the molar ratio of the quercetin to the ergothioneine is 1:1-1:10. Preferably, the molar ratio of the quercetin to the ergothioneine is 1:2.

[0041] In some embodiments, the first solvent is one or more of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution; the second solvent is one of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution.

[0042] In some embodiments, the concentration of the quercetin solution is 1-10 mol / L; the concentration of the ergothioneine is 1-10 mol / L. In a preferred embodiment, the concentration of the quercetin solution is 1 mol / L; the concentration of the ergothioneine is 2 mol / L.

[0043] In some embodiments, the inert atmosphere is nitrogen or argon.

[0044] In some embodiments, the temperature of the ionized salt-forming reaction is 30-100 DEG C, and the time of the ionized salt-forming reaction is 4-24h. Quercetin is used as an anion precursor and ergothioneine is used as a cationic precursor, and supramolecular quercetin-ergothioneine is synthesized by ionized salt-forming reaction. The supramolecular quercetin-ergothioneine has the biological activity and efficacy of both quercetin and ergothioneine, and greatly improves the solubility and bioavailability of quercetin and ergothioneine, and enhances its application effect.

[0045] At the reaction temperature, the reaction rate is relatively fast and the occurrence of side reactions can be effectively suppressed; and at the reaction time, the complete reaction between quercetin and ergothioneine can be promoted.

[0046] In some embodiments, the step of concentrating the solution after the reaction and performing reduced pressure distillation and crystallization comprises: rotary evaporating the solution after the reaction of quercetin and ergothioneine under vacuum conditions, and when the volume of the solution is concentrated to 1 / 10-1 / 5 of the original volume, performing reduced pressure distillation and crystallization.

[0047] In some embodiments, the drying method is vacuum drying, the drying temperature is 30-100° C., and the drying time is 24-48 hours. The drying is performed to completely remove the remaining water or solvents such as ethanol in the product.

[0048] The embodiment of the present invention also provides a supramolecule constructed by quercetin and ergothioneine, which is prepared by the preparation method of the supramolecule constructed by quercetin and ergothioneine.

[0049] The supermolecule constructed by quercetin and ergothioneine is assembled by combining quercetin molecules and ergothioneine molecules through non-covalent bonding.

[0050] The supramolecular quercetin-ergothioneine of the present invention has the following effects: firstly, compared with single quercetin, the supramolecular quercetin-ergothioneine has better biocompatibility, and significantly improves its transdermal absorption, thereby enhancing the application effect; and secondly, the supramolecular quercetin-ergothioneine completely retains the main molecular skeleton of the quercetin monomer and the characteristics of ergothioneine, thereby having the biological activities and effects of the two substances at the same time.

[0051] The embodiment of the present invention provides an application of the supramolecule constructed by quercetin and ergothioneine in preparing cosmetics or drug carriers.

[0052] The supramolecular quercetin-ergothioneine can be used as a raw material for the formulation of medicines and cosmetics, and can also be used as a raw material for carriers of drugs such as transdermal delivery.

[0053] The following describes it in detail through specific embodiments.

[0054] Example 1

[0055] The schematic flow diagram of the preparation method of the supramolecule constructed by quercetin and ergothioneine is as follows Figure 1 The specific preparation steps are as follows:

[0056] Under a nitrogen atmosphere, 0.01 mol of quercetin was dissolved in 10 mL of water and ethanol to obtain a quercetin solution in a reactor, 0.02 mol of ergothioneine was dissolved in 10 mL of water to obtain an ergothioneine solution, and the ergothioneine solution was dropwise added to the reactor of the quercetin solution, heated to 60 ° C, and ionized salt was reacted for 12 hours;

[0057] After the reaction is completed, the solution is concentrated to 1 / 10 of the reaction solution under vacuum conditions, and the product is crystallized by distillation under reduced pressure, and the supermolecular quercetin-ergothioneine crystalline product is obtained by filtering and washing, and the supermolecular quercetin-ergothioneine with a purity exceeding 99% is obtained by drying in a vacuum drying oven for 48 hours, and the yield is 97.35%, thereby obtaining the supermolecular quercetin-ergothioneine.

[0058] At room temperature, quercetin and ergothioneine were mixed in a molar ratio of 1:1, and a magnetic stirrer was used to fully mix the mixed solution. The system was observed to be completely dissolved to a clear state, thereby obtaining a supramolecular quercetin-ergothioneine solution.

[0059] like Figure 2 As shown, the nuclear magnetic proton spectrum data of supramolecular quercetin-ergothioneine obtained in this embodiment are: 1 HNMR (400MHz, DMSO) δ12.93 (s, 1H), 12.48 (d, 1H), 12.00 (s, 1H), 11.77 (s, 1H), 10.89 (s, 1H), 9.66 (s, 1H), 9.36 (d, 1H), 7.71 (t, 1H), 7.54 (dd, 1H), 6.85 (dd, 1H), 6.51 (m, 1H), 6.44 (m, 1H), 6.20 (d, 1H), 1.16 (d, 3H), 1.01 (m, 2H); NMR carbon spectrum data are: 13C NMR (400MHz, DMSO) δ176.29, 167.16, 164.39, 161.16, 160.74, 156.59, 148.18, 147.25, 145.52, 136.18, 12 4.10,122.29,120.42,116.06,115.79,115.51,113.57,103.44,98.65,93.81,75.20,56.04,51.70,22.84.

[0060] The melting point of the supramolecular quercetin-ergothioneine prepared in this embodiment is 219°C.

[0061] like Figure 3 As shown, it can be seen from the two-dimensional NMR spectrum that quercetin and ergothioneine can form supramolecules through intermolecular forces.

[0062] Example 2

[0063] The preparation method of supramolecular quercetin-ergothioneine (molar ratio 1:1) of the present embodiment comprises the following steps: under a nitrogen atmosphere, dissolving 0.01 mol of quercetin in a reactor with 10 mL of water and ethanol, dissolving 0.01 mol of ergothioneine with 10 mL of water, and adding dropwise to the reactor in which quercetin is dissolved, heating to 60 ° C, and ionizing and salting reacting for 12 hours;

[0064] After the reaction is completed, the solution is concentrated to 1 / 10 of the reaction solution under vacuum conditions, and the product is crystallized by distillation under reduced pressure, and the supermolecular quercetin-ergothioneine crystalline product is obtained by filtering and washing, and the supermolecular quercetin-ergothioneine with a purity exceeding 99% is obtained by drying in a vacuum drying oven for 48 hours, and the yield is 97.35%, thereby obtaining the supermolecular quercetin-ergothioneine.

[0065] At room temperature, quercetin and ergothioneine were mixed in a molar ratio of 1:1, and a mixed solution was obtained by fully mixing with a magnetic stirrer. The system was observed to be completely dissolved to a clear state to obtain a quercetin-ergothioneine solution, and then the solvent was removed by a rotary evaporator to obtain supramolecular quercetin-ergothioneine.

[0066] like Figure 4 As shown, the nuclear magnetic proton spectrum data of supramolecular quercetin-ergothioneine obtained in this embodiment are: 1HNMR (400MHz, DMSO) δ12.14 (s, 1H), 11.96 (d, 1H), 11.79 (s, 1H), 8.21 (s, 1H), 6.97 (s, 1H), 6.64 (s, 1H), 6.58 (d, 1H), 3.87 (t, 1H), 3.24 (dd, 1H), 3.04 (dd, 1H), 2.68 (m, 1H), 1.29 (m, 1H), 1.14 (d, 1H), 0.85 (d, 3H); NMR carbon spectrum data are: 13 C NMR (400MHz, DMSO) δ176.16, 166.34, 163.63, 160.66, 159.75, 156.77, 148.19, 144.88, 135 .02,124.78,122.48,120.41,115.52,113.48,102.64,96.88,93.23,76.37,51.63,23.55.

[0067] The melting point of the supramolecular quercetin-ergothioneine prepared in this embodiment is 207°C.

[0068] like Figure 5 As shown, from two-dimensional nuclear magnetic spectrum, it can be seen that supramolecular quercetin-ergothioneine can be formed by intermolecular force between quercetin and thioneine (mol ratio is 1: 1), but the force between quercetin and thioneine in Example 2 is much weaker than that in supermolecular quercetin-ergothioneine (mol ratio is 1: 2) in Example 1. Therefore, it is better to prepare the quercetin used in supramolecular quercetin-ergothioneine with a mol ratio of 1: 2.

[0069] The Franz diffusion cell was used to detect the content after incubation for different time periods using a pig skin model to evaluate the penetration effect of the supramolecular quercetin-ergothioneine prepared in Example 1. Figure 6 As shown, it can be seen from the transdermal efficiency diagram that as the transdermal time is prolonged, the transdermal efficiency of supramolecular quercetin-ergothioneine also gradually increases until a balance state is reached. The transdermal efficiency of supramolecular quercetin-ergothioneine is 18.03 times that of single quercetin and 7.53 times that of ergothioneine. This is because the structure of supramolecular quercetin-ergothioneine contains the force of covalent bonds, which can increase the combination of supramolecular and skin, thereby promoting its penetration.

[0070] After the supramolecular quercetin-ergothioneine prepared in Example 1 was co-incubated with keratinocytes in a cell culture incubator for 24 hours, the effect of supramolecular quercetin-ergothioneine on keratinocyte viability was detected by an ELISA instrument. Figure 7 As shown in the cell viability graph, it can be seen that the survival rate of keratinocytes treated with supramolecular quercetin-ergothioneine is over 90%.

[0071] like Figure 8 Shown, from the solubility diagram, it can be seen that the solubility of the supramolecular quercetin-ergothioneine prepared in Example 1 is improved by about 49.28 times in aqueous solution than single quercetin, and is improved by about 3.96 times in ethanol solution. This is because the covalent bond in supramolecular quercetin-ergothioneine can promote it to dissolve in solution, thereby improving its solubility.

[0072] In summary, the present invention uses ergothioneine as a cationic precursor and quercetin as an anionic precursor, and realizes the separation and purification of the product by concentrated crystallization after ionized salt-forming reaction, and obtains the supramolecular constructed by quercetin and ergothioneine, i.e. supramolecular quercetin-ergothioneine. The solubility of supramolecular quercetin-ergothioneine is improved by about 49 times than that of single quercetin in aqueous solution, and supramolecular quercetin-ergothioneine has the antioxidant, anti-inflammatory, soothing and repairing effects and biological activity of quercetin and ergothioneine at the same time.

[0073] It should be understood that the application of the present invention is not limited to the above examples. For ordinary technicians in this field, improvements or changes can be made based on the above description. All these improvements and changes should fall within the scope of protection of the claims attached to the present invention.

Claims

1. A method for preparing a supramolecule constructed of quercetin and ergothioneine, characterized in that, Includes steps: dissolving quercetin in a first solvent to obtain a quercetin solution; Dissolving thioneine in a second solvent to obtain an thioneine solution; Under an inert atmosphere, the thioneine solution is added to the quercetin solution and mixed, and an ionized salt-forming reaction is carried out under heating conditions; The solution after the reaction is concentrated, distilled and crystallized, and then filtered, washed and dried to obtain the supramolecule constructed by quercetin and ergothioneine.

2. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct, is characterized in that, The molar ratio of the quercetin to the ergothioneine is 1:1-1:

10.

3. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct, is characterized in that, The first solvent is one or more of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution; the second solvent is one of ethanol, water, methanol, isopropanol, acetone, propylene glycol, butylene glycol and pentanediol solution.

4. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct, is characterized in that, The concentration of the quercetin solution is 1-10 mol / L; the concentration of the ergothioneine is 1-10 mol / L.

5. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct, is characterized in that, The inert atmosphere is nitrogen or argon.

6. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct is characterized in that, The temperature of the ionization salt-forming reaction is 30-100° C., and the time of the ionization salt-forming reaction is 4-24 hours.

7. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct, is characterized in that, The step of concentrating the solution after the reaction and performing reduced pressure distillation and crystallization comprises: performing rotary evaporation on the solution after the reaction of quercetin and ergothioneine under vacuum conditions, and performing reduced pressure distillation and crystallization when the volume of the solution is concentrated to 1 / 10-1 / 5 of the original volume.

8. the preparation method of the supramolecule that quercetin according to claim 1 and thioneine construct is characterized in that, The drying method is vacuum drying, the drying temperature is 30-100° C., and the drying time is 24-48 hours.

9. A supramolecule constructed of quercetin and ergothioneine, characterized in that, Prepared by the preparation method of the supramolecule constructed by quercetin and ergothioneine according to any one of claims 1 to 8; And / or, the supramolecule constructed by quercetin and ergothioneine is assembled by combining quercetin molecules and ergothioneine molecules through non-covalent bonding.

10. Use of the supramolecule constructed by quercetin and ergothioneine according to claim 9 in preparing cosmetics or drug carriers.

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