Ternary supramolecular and preparation method thereof, ternary supramolecular alcohol body and preparation method thereof and application thereof

Abstract

The present invention provides a ternary supramolecule and a preparation method thereof, a ternary supramolecule alcohol body and a preparation method and application thereof, and specifically relates to the technical field of biomedical materials. The ternary supramolecule includes xylitol, betaine and olive leaf extract; the xylitol and the betaine constitute a binary supramolecular solution, and the olive leaf extract is dissolved in the binary supramolecular solution. The ternary supramolecule provided by the present invention stabilizes the olive leaf extract in a binary supramolecular solution composed of xylitol and betaine in a supramolecular form, and the formed ternary supramolecule has a significant protective effect on oleuropein, preventing oleuropein from being degraded in water-based products. In the ternary supramolecule, oleuropein-betaine-xylitol form a ternary eutectic, and π-π stacking is formed inside, causing electron cloud rearrangement, reducing the exposure activity of the polyphenols that are easily attacked by oxygen, thereby protecting them from degradation.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedical materials, and in particular to a ternary supramolecular and a preparation method thereof, a ternary supramolecular alcohol body and a preparation method and application thereof. Background Art

[0002] Olive leaf extract is a 100% natural water-soluble active ingredient extracted from olive leaves. Olive (Olea europaea L.) is a plant of the genus Oleaceae, mostly produced in the Mediterranean coastal areas of southern Europe, and is a Mediterranean subtropical tree species. Modern pharmacological research has shown that compounds such as oleuropein, hydroxytyrosol, polyphenols, and flavonoids in olive leaves have multiple pharmacological activities such as anti-inflammatory and antibacterial, antioxidant, antiviral, anti-tumor, and hypoglycemic and blood lipid lowering, and are widely used in the field of topical preparations.

[0003] The highest content of active ingredients in olive leaf extract is oleuropein, which is a secoiridoid ether terpenoid compound with strong antioxidant properties of antibacterial and anti-inflammatory properties, but its stability is very poor. It is very easy to degrade and deteriorate under the interference of external environment such as light, heat, oxygen, pH, enzymes, etc., which is specifically manifested as darkening of color and obvious decrease in content, so it is very difficult to be applied to conventional topical preparations. In addition, the main active ingredients in olive leaf extract, such as oleuropein, are all water-soluble components, with low logP (oil-water partition coefficient), weak fat solubility, and it is difficult to penetrate the lipid stratum corneum barrier composed of ceramide, cholesterol ester, and fatty acids, and the penetration ability is weak, so the bioavailability is low.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] One of the objectives of the present invention is to provide a ternary supramolecule to solve at least one of the above technical problems in the prior art.

[0006] A second object of the present invention is to provide a method for preparing a ternary supramolecule.

[0007] The third object of the present invention is to provide a ternary supramolecular alcohol body.

[0008] A fourth object of the present invention is to provide a method for preparing a ternary supramolecular alcohol body.

[0009] A fifth object of the present invention is to provide an application of a ternary supramolecular alcohol body.

[0010] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0011] The first aspect of the present invention provides a ternary supramolecule comprising xylitol, betaine and olive leaf extract;

[0012] The xylitol and the betaine constitute a binary supramolecular solution, and the olive leaf extract is dissolved in the binary supramolecular solution.

[0013] Furthermore, in the binary supramolecular solution, the molar ratio of betaine to xylitol is 1:1 to 1:3.

[0014] Preferably, in the ternary supramolecule, the mass ratio of the binary supramolecule solution to the olive leaf extract is 1:1 to 2.2.

[0015] Preferably, the content of oleuropein in the olive leaf extract is greater than 80%.

[0016] Preferably, the content of oleuropein in the olive leaf extract is ≥85%.

[0017] Preferably, the content of oleuropein in the olive leaf extract is ≥90%.

[0018] The second aspect of the present invention provides a method for preparing the ternary supramolecule, comprising the following steps:

[0019] A. Add betaine and xylitol into a container, mix them evenly at 120°C to 150°C to obtain a clear liquid, and continue to cool to room temperature to obtain a binary supramolecular solution;

[0020] B. dissolving the olive leaf extract in the binary supramolecular solution to obtain the ternary supramolecular solution.

[0021] Furthermore, in step A, the mixing is performed under vacuum conditions.

[0022] Preferably, the vacuum degree of the vacuum condition is ≤-0.05MPa.

[0023] Preferably, in step A, the mixing time is 3 h to 10 h.

[0024] Preferably, in step B, the dissolving temperature is 30°C to 50°C.

[0025] The third aspect of the present invention provides a ternary supramolecular alcohol body comprising a ternary supramolecular, lecithin, glycerol, alcohol and deionized water.

[0026] The ternary supramolecule is the ternary supramolecule described in the first aspect;

[0027] Alternatively, the ternary supramolecule is prepared by the preparation method described in the second aspect.

[0028] Furthermore, in terms of mass percentage, the ternary supramolecular alcohol body includes 12.5% ​​to 26.5% of ternary supramolecular, 4.0% to 10.0% of lecithin, 30.0% to 40.0% of glycerol, 15.0% to 25.0% of alcohol, and the remainder is deionized water.

[0029] Preferably, the lecithin is soy lecithin.

[0030] Preferably, the soybean lecithin has a phosphatidylcholine content of ≥80%.

[0031] Preferably, the phosphatidylcholine content in the soybean lecithin is ≥ 90%.

[0032] Preferably, the alcohol is a small molecule alcohol.

[0033] Preferably, the carbon number of the small molecule alcohol is ≤5.

[0034] Preferably, the small molecule alcohol is 1,3-propylene glycol.

[0035] Furthermore, the particle size of the ternary supramolecular alcohol body is 20nm to 100nm;

[0036] Preferably, the particle size of the ternary supramolecular alcohol body is 20nm to 50nm.

[0037] The fourth aspect of the present invention provides a method for preparing the ternary supramolecular alcohol body, comprising the following steps:

[0038] a, first dissolving the lecithin in the alcohol to obtain a lecithin solution;

[0039] b. adding the ternary supramolecule into the lecithin solution to obtain the first colostrum;

[0040] c. uniformly mixing the glycerol and the deionized water to obtain an external water phase;

[0041] d. adding the first colostrum to the external aqueous phase and mixing them evenly to obtain the second colostrum;

[0042] e. Homogenize the second colostrum to obtain the ternary supramolecular ethosome.

[0043] Furthermore, in step a, the dissolving temperature is 50°C to 70°C.

[0044] Preferably, in step b, the temperature of the lecithin solution is 40°C to 60°C.

[0045] Preferably, in step c, the mixing temperature is 40°C to 60°C.

[0046] Preferably, in step d, the temperature of the external aqueous phase is 40°C to 60°C.

[0047] Preferably, in step e, the homogenization pressure is 10000psi to 20000psi.

[0048] Preferably, the number of cycles of the homogenization is 2 to 6 times.

[0049] Preferably, the homogenized discharge temperature is 8°C to 25°C.

[0050] The fifth aspect of the present invention provides the use of a ternary supramolecular ethosome in an external preparation, wherein the ternary supramolecular ethosome is the ternary supramolecular ethosome described in the third aspect;

[0051] or,

[0052] The ternary supramolecular alcohol body is prepared by the preparation method described in the fourth aspect.

[0053] Compared with the prior art, the present invention has at least the following beneficial effects:

[0054] The ternary supramolecule provided by the present invention stabilizes the olive leaf extract in a binary supramolecule solution composed of xylitol and betaine in a supramolecule form, and the formed ternary supramolecule has a significant protective effect on oleuropein, preventing the degradation of oleuropein in water-based products. In the ternary supramolecule, oleuropein-betaine-xylitol forms a ternary eutectic, and π-π stacking is formed inside, causing electron cloud rearrangement, reducing the exposure activity of the polyphenols that are easily attacked by oxygen, thereby protecting them from degradation. The ternary supramolecule has the characteristics of high skin absorption rate, good stability, and convenient application, and significantly improves the application effect of the olive leaf extract.

[0055] The preparation method of the ternary supramolecule provided by the present invention adopts a two-step method to first form a binary supramolecule solution with betaine and xylitol at high temperature, so that the ternary supramolecule can be prepared under low temperature conditions. The degradation of the thermally unstable component olive leaf extract due to severe heating is avoided, and in particular, oleuropein is prevented from being degraded into hydroxytyrosol, which will rapidly degrade, turn yellow and deteriorate, produce toxic and harmful degradation products, and affect product quality. The two-step operation method for preparing the ternary supramolecule greatly improves the stability of the product and avoids the damage of the unstable component to high temperature.

[0056] The ternary supramolecular alcohol body provided by the present invention has good stability and permeability, significantly improves the skin absorption rate of oleuropein, and improves its bioavailability. The ternary supramolecular oleuropein-betaine-xylitol ternary supramolecular hydrophilicity is stronger, and is wrapped in the inner water phase by the phospholipid bilayer, which further reduces the possibility that oleuropein is stimulated by the external environment and thus unstable. Furthermore, lecithin has good skin affinity and is more likely to form a phospholipid bilayer structure similar to the skin cell membrane, making it easier to carry oleuropein and penetrate into the deep layer of the skin, giving full play to its beneficial effects.

[0057] The preparation method of the ternary supramolecular alcohol body provided by the present invention is simple, and no toxic or harmful organic solvents are introduced during the process, and no chemical penetration enhancers such as azone that are not welcome in cosmetics are introduced, and no solubilizers or surfactants are introduced, thereby achieving safe and effective application of olive leaf extract.

[0058] The application of the ternary supramolecular alcohol body provided by the present invention provides an oleuropein product with better performance for external preparations, improves the stability and permeability of olive leaf extract, significantly improves the skin absorption rate and bioavailability of olive leaf extract, and reduces the dosage and cost of olive leaf extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0060] Figure 1 The particle size test results of the ternary supramolecular alcohol body of Example 9;

[0061] Figure 2 This is a comparison chart of the retention test obtained in Test Example 2;

[0062] Figure 3 The final appearance of the product of Example 9 was examined at different temperatures for 30 days;

[0063] Figure 4 The final appearance of the product of Comparative Example 1 was examined at different temperatures for 30 days;

[0064] Figure 5 The final appearance of the product of Comparative Example 2 was examined at different temperatures for 30 days;

[0065] Figure 6 is the infrared spectrum of the binary supramolecule obtained in Example 1;

[0066] Figure 7 is the infrared spectrum of the ternary supramolecule obtained in Example 1;

[0067] Figure 8 This is the mass spectrum of the ternary supramolecule obtained in Example 1. DETAILED DESCRIPTION

[0068] The embodiments of the present invention will be described in detail below in conjunction with the embodiments and examples, but those skilled in the art will understand that the following embodiments and examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0069] The first aspect of the present invention provides a ternary supramolecule comprising xylitol, betaine and olive leaf extract;

[0070] The xylitol and the betaine constitute a binary supramolecular solution, and the olive leaf extract is dissolved in the binary supramolecular solution.

[0071] The ternary supramolecule provided by the present invention stabilizes the olive leaf extract in a binary supramolecule solution composed of xylitol and betaine in a supramolecule form, and the formed ternary supramolecule has a significant protective effect on oleuropein, preventing the degradation of oleuropein in water-based products. In the ternary supramolecule, oleuropein-betaine-xylitol form a ternary eutectic, and a π-π stack is formed inside, causing electron cloud rearrangement, reducing the exposure activity of the part of the polyphenol that is easily attacked by oxygen, thereby protecting it from degradation.

[0072] Supramolecular molecules usually refer to two or more molecules that are combined together by intermolecular interactions to form complex, organized aggregates that maintain a certain integrity and have clear microstructures and macroscopic properties.

[0073] The main component of olive leaf extract is oleuropein, which is extremely unstable and rapidly degrades and turns yellow in aqueous solution, and its activity decreases, so it cannot be widely used in conventional external preparations such as lotions, emulsions, creams, essences, and facial masks. The present invention prepares the olive leaf extract into a supramolecular form, making it stable and non-discoloring, and more convenient to use.

[0074] Furthermore, in the binary supramolecular solution, the molar ratio of betaine to xylitol is 1:1 to 1:3.

[0075] Within the above molar ratio range, xylitol and betaine can form a binary supramolecular solution. If the molar ratio of betaine to xylitol is too high or too low, it is not conducive to the formation of a binary supramolecular liquid, and a binary supramolecular solution that is liquid at room temperature cannot be prepared, which will result in the inability to dissolve the olive leaf extract at room temperature or under slightly heated conditions, and the olive leaf extract will degrade and discolor when dissolved at high temperature.

[0076] Typically but not limiting, the molar ratio of betaine to xylitol may be, for example, 1:1, 1:2 or 1:3, or any value within the range of 1:1-3.

[0077] Preferably, in the ternary supramolecule, the mass ratio of the binary supramolecule solution to the olive leaf extract is 1:1 to 2.2.

[0078] The mass ratio of the binary supramolecular solution to the olive leaf extract is less than 1:2.2, and the amount of the olive leaf extract is too large, making it difficult to dissolve the liquid ternary supramolecular solution at room temperature, and thus the subsequent process operation cannot be performed. When the mass ratio of the binary supramolecular solution to the olive leaf extract is greater than 1:1, the binary supramolecular solution is too large, which will cause the ternary supramolecular to be unstable, mainly because the stability between the ternary supramolecular is through hydrogen bonding and π-π stacking, and oleuropein provides the main source of π-π stacking, so its content plays a very important role in the stability of the ternary supramolecular. At the same time, due to the effect of hydrogen bonding and π-π stacking, the electron cloud is rearranged, and the unstable groups in oleuropein are protected, thereby achieving the improvement of stability.

[0079] The olive leaf extract used in the present invention can be extracted by anhydrous alcoholysis, enzyme destruction, leaching, ultrasonic assisted extraction, microwave assisted extraction, supercritical carbon dioxide extraction or the like, or an optimized combination of multiple methods, and the extraction method is not particularly limited. The extract can be a solid powder or other states, and the state of the olive leaf extract is not particularly limited.

[0080] Preferably, the content of oleuropein in the olive leaf extract is greater than 80%.

[0081] Preferably, the content of oleuropein in the olive leaf extract is ≥85%.

[0082] Preferably, the content of oleuropein in the olive leaf extract is ≥90%.

[0083] The second aspect of the present invention provides a method for preparing the ternary supramolecule, comprising the following steps:

[0084] A. Add betaine and xylitol into a container, mix them evenly at 120°C to 150°C to obtain a clear liquid, and continue to cool to room temperature to obtain a binary supramolecular solution;

[0085] B. dissolving the olive leaf extract in the binary supramolecular solution to obtain the ternary supramolecular solution.

[0086] The preparation method of the ternary supramolecule provided by the present invention adopts a two-step method to first form a binary supramolecule solution with betaine and xylitol at high temperature, so that the ternary supramolecule can be prepared under low temperature conditions. The degradation of the thermally unstable component olive leaf extract due to severe heating is avoided, and in particular, oleuropein is prevented from being degraded into hydroxytyrosol, which will rapidly degrade, turn yellow and deteriorate, produce toxic and harmful degradation products, and affect product quality. The two-step operation method for preparing the ternary supramolecule greatly improves the stability of the product and avoids the damage of the unstable component to high temperature.

[0087] It should be noted that the binary supramolecular solution is a transparent viscous liquid at room temperature; the ternary supramolecular solution is a light yellow transparent viscous liquid at room temperature.

[0088] In step A, betaine and xylitol can form a deep eutectic solvent at 120°C to 150°C. When the temperature is lower than 120°C, it is difficult to form a binary supramolecule because the melting point of betaine is relatively high; when the temperature is higher than 150°C, it is not conducive to stability because betaine and xylitol are both organic substances.

[0089] Typically but not limiting, the mixing temperature may be, for example, 120°C, 125°C, 130°C, 135°C, 140°C, 145°C or 150°C, or any value within the range of 120°C to 150°C.

[0090] Furthermore, in step A, the mixing is carried out under vacuum conditions. Under vacuum conditions, the melting points of the two solids will decrease, and the viscosity will decrease, so the operation can be performed at a temperature below 150°C.

[0091] Preferably, the vacuum degree of the vacuum condition is ≤-0.05MPa.

[0092] Preferably, in step A, the mixing time is 3 h to 10 h.

[0093] In step A, if the mixing time is less than 3 hours, the binary supramolecular solution is easy to separate; if the mixing time is greater than 10 hours, it will lead to increased energy consumption, resulting in an increase in preparation cost and reduced production efficiency.

[0094] Typically but not limiting, the mixing time may be, for example, 3 h, 4 h, 5 h, 6 h, 7 h, 8 h, 9 h or 10 h, or any value within the range of 3 h to 10 h.

[0095] Preferably, in step B, the dissolving temperature is 30°C to 50°C.

[0096] In step B, if the dissolution temperature is less than 30°C, the viscosity of the system is too high and stirring is impossible; if the dissolution temperature is greater than 50°C, the active substance is at risk of degradation, affecting the safety and effectiveness of the product.

[0097] Typically but not limiting, the dissolution temperature may be, for example, 30°C, 35°C, 40°C, 45°C or 50°C, or any value within the range of 30°C to 50°C.

[0098] The third aspect of the present invention provides a ternary supramolecular alcohol body comprising a ternary supramolecular, lecithin, glycerol, alcohol and deionized water.

[0099] The ternary supramolecule is the ternary supramolecule described in the first aspect;

[0100] Alternatively, the ternary supramolecule is prepared by the preparation method described in the second aspect.

[0101] The ternary supramolecular alcohol body provided by the present invention has good stability and permeability, significantly improves the skin absorption rate of oleuropein, and improves its bioavailability. The ternary supramolecular oleuropein-betaine-xylitol ternary supramolecular hydrophilicity is stronger, and is wrapped in the inner water phase by the phospholipid bilayer, which further reduces the possibility that oleuropein is stimulated by the external environment and thus unstable. Furthermore, lecithin has good skin affinity and is more likely to form a phospholipid bilayer structure similar to the skin cell membrane, making it easier to carry oleuropein and penetrate into the deep layer of the skin, giving full play to its beneficial effects.

[0102] Ethosomes are phospholipid vesicles containing relatively high concentrations of alcohol, which are composed of phospholipids, alcohol and water. This carrier has high biocompatibility due to its special structure, can improve the bioavailability of olive leaf extract, and is an ideal olive leaf extract delivery carrier. The bilayer fluidity of ethosomes is high, easy to deform, can penetrate the skin barrier, significantly enhance the penetration ability of drugs to the skin, break through the barrier of the stratum corneum, promote olive leaf extract to penetrate the skin, increase the accumulation of olive leaf extract in the skin, and can further carry olive leaf extract into cells. In addition, ethosomes can also promote the transmission and penetration of olive leaf extract, further improve the transdermal absorption effect.

[0103] Furthermore, in terms of mass percentage, the ternary supramolecular alcohol body includes 12.5% ​​to 26.5% of ternary supramolecular, 4.0% to 10.0% of lecithin, 30.0% to 40.0% of glycerol, 15.0% to 25.0% of alcohol, and the remainder is deionized water.

[0104] Preferably, in the ternary supramolecular ethosome, the mass ratio of phospholipid to ternary supramolecular is 1:1-3.

[0105] Preferably, in the ternary supramolecular alcohol body, the mass ratio of lecithin to 1,3-propylene glycol is 1:2.5-4.

[0106] Typically but not limiting, in the ternary supramolecular alcohol body, the mass percentage of the ternary supramolecular can be, for example, 12.5%, 13%, 15%, 18%, 20%, 23% or 26.5%, or any value within the range of 12.5% ​​to 26.5%; the mass percentage of lecithin can be, for example, 4.0%, 5.0%, 6.0%, 7.0%, 8.0%, 9.0% or 10.0%, or any value within the range of 4.0% to 10.0%; The mass percentage of oil can be, for example, 30.0%, 32.0%, 34.0%, 36.0%, 38.0% or 40.0%, or any value within the range of 30.0% to 40.0%; the mass percentage of alcohol can be, for example, 15.0%, 17.0%, 19.0%, 21.0%, 23.0% or 25.0%, or any value within the range of 15.0% to 25.0%; the balance being deionized water means that deionized water is used to make up 100%.

[0107] Preferably, the lecithin is soy lecithin.

[0108] Soy lecithin is a high-purity non-hydrogenated lecithin with good skin affinity. It is easier to form a phospholipid bilayer structure similar to the skin cell membrane, making it easier for it to carry oleuropein and penetrate into the deep layers of the skin to fully exert its beneficial effects.

[0109] Preferably, the soybean lecithin has a phosphatidylcholine content of ≥80%.

[0110] Preferably, the phosphatidylcholine content in the soybean lecithin is ≥90%. High-purity soybean lecithin can easily prepare double-layer structure liposomes with soft deformability, which is beneficial to the absorption of olive leaf extract.

[0111] Preferably, the alcohol is a small molecule alcohol.

[0112] Preferably, the carbon number of the small molecule alcohol is ≤5.

[0113] Typically, but not limiting, the small molecule alcohol may be, for example, ethanol, propanol, 1,3-propylene glycol, butanol or pentanol.

[0114] Preferably, the small molecule alcohol is 1,3-propylene glycol.

[0115] Furthermore, the particle size of the ternary supramolecular alcohol body is 20nm to 100nm. Within this particle size range, the ternary supramolecular alcohol body has a smaller specific surface area, which significantly increases the contact area between the active substance and the skin and increases the penetration effect.

[0116] Preferably, the particle size of the ternary supramolecular alcohol body is 20nm to 50nm.

[0117] The fourth aspect of the present invention provides a method for preparing the ternary supramolecular alcohol body, comprising the following steps:

[0118] a, first dissolving the lecithin in the alcohol to obtain a lecithin solution;

[0119] b. adding the ternary supramolecule into the lecithin solution to obtain the first colostrum;

[0120] c. uniformly mixing the glycerol and the deionized water to obtain an external water phase;

[0121] d. adding the first colostrum to the external aqueous phase and mixing them evenly to obtain the second colostrum;

[0122] e. Homogenize the second colostrum to obtain the ternary supramolecular ethosome.

[0123] The preparation method of the ternary supramolecular alcohol body provided by the present invention is simple, and no toxic or harmful organic solvents are introduced during the process, and no chemical penetration enhancers such as azone that are not welcome in cosmetics are introduced, and no solubilizers or surfactants are introduced, thereby achieving safe and effective application of olive leaf extract.

[0124] Furthermore, in step a, the dissolving temperature is 50°C to 70°C.

[0125] Typically but not limiting, in step a, the dissolving temperature may be, for example, 50°C, 55°C, 60°C, 65°C or 70°C, or any value within the range of 50°C to 70°C.

[0126] Preferably, in step b, the temperature of the lecithin solution is 40°C to 60°C.

[0127] Typically but not limiting, in step b, the temperature of the lecithin solution can be, for example, 40°C, 45°C, 50°C, 55°C or 60°C, or any value within the range of 40°C to 60°C.

[0128] Preferably, in step c, the mixing temperature is 40°C to 60°C.

[0129] Typically but not limiting, in step c, the mixing temperature may be, for example, 40°C, 45°C, 50°C, 55°C or 60°C, or any value within the range of 40°C to 60°C.

[0130] Preferably, in step d, the temperature of the external aqueous phase is 40°C to 60°C.

[0131] Typically but not limiting, in step d, the temperature of the external aqueous phase can be, for example, 40°C, 45°C, 50°C, 55°C or 60°C, or any value within the range of 40°C to 60°C.

[0132] Preferably, in step e, the homogenization pressure is 10000psi to 20000psi.

[0133] Preferably, the number of cycles of the homogenization is 2 to 6 times.

[0134] Preferably, the homogenized discharge temperature is 8°C to 25°C.

[0135] The fifth aspect of the present invention provides the use of a ternary supramolecular ethosome in an external preparation, wherein the ternary supramolecular ethosome is the ternary supramolecular ethosome described in the third aspect;

[0136] or,

[0137] The ternary supramolecular alcohol body is prepared by the preparation method described in the fourth aspect.

[0138] The application of the ternary supramolecular alcohol body provided by the present invention provides an oleuropein product with better performance for external preparations, improves the stability and permeability of olive leaf extract, significantly improves the skin absorption rate and bioavailability of olive leaf extract, and reduces the dosage and cost of olive leaf extract.

[0139] Preferably, the external preparation comprises an external skin care product.

[0140] Preferably, the external skin care products include conventional cosmetic dosage forms such as lotion, emulsion, essence, facial mask, cream, etc.

[0141] It should be noted that the amount of the ternary supramolecular alcohol body added in external skin care products is 0.01wt.% to 20wt.%, and the preferred amount is 1wt.% to 10wt.%, which has better stability and use effect.

[0142] In the preparation of external skin care products, the addition temperature of the ternary supramolecular alcohol body is controlled below 40°C and is added during the cold preparation stage of the formula.

[0143] The present invention is further described below by specific examples and comparative examples, but it should be understood that these examples are only used for more detailed description and should not be understood as limiting the present invention in any form. The raw materials used in the examples and comparative examples of the present invention, if no specific conditions are specified, are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially.

[0144] Examples 1-8

[0145] These embodiments provide a ternary supramolecule, and the specific raw material ratios are shown in Table 1 below.

[0146] Table 1

[0147]

[0148] The preparation method of the ternary supramolecule of the above embodiment is as follows:

[0149] 1. Add betaine and xylitol to a clean container, stir continuously for 6 hours at 130°C under vacuum conditions (vacuum degree = -0.06Mpa) to obtain a clear liquid, and cool to room temperature to obtain a transparent viscous liquid, which is a binary supramolecular solution.

[0150] 2. At 45° C., dissolve olive leaf extract (oleuropein content ≥ 90%) in the binary supramolecular solution obtained in step 1, and cool to room temperature to obtain a light yellow transparent viscous liquid, which is a ternary supramolecular solution.

[0151] During the preparation process of Examples 3 and 4, the binary supramolecules obtained were in solid state and the preparation was unsuccessful; the ternary supramolecules prepared in Example 7 were unstable during storage and precipitated; the olive leaf extract in Example 8 could not be dissolved in the binary supramolecule solution and the preparation was unsuccessful.

[0152] Examples 9-12

[0153] These embodiments provide a ternary supramolecular alcohol body, including ternary supramolecular, 8.0g of lecithin, 20.0g of 1,3-propylene glycol, 35.0g of glycerol, and 100g of deionized water. The ternary supramolecular in Examples 9-12 correspond to those provided in Examples 1, 2, 5 and 6, respectively, and the amounts used are the total yields obtained in Examples 1, 2, 5 and 6, respectively. The specific raw material ratios are shown in Table 2 below.

[0154] Table 2

[0155]

[0156] The preparation method of ternary supramolecular alcohol body is as follows:

[0157] 1. Add lecithin (phosphatidylcholine content ≥ 90%) to 1,3-propylene glycol and dissolve it at 60°C to obtain a transparent liquid, which is the lecithin solution.

[0158] 2. At 50°C and under continuous stirring, the ternary supramolecules were slowly injected into the lecithin solution to obtain the first colostrum.

[0159] 3. Dissolve glycerol and pure water in a clean container under stirring at 50°C water bath to obtain an external aqueous phase. Then, inject the first colostrum into the external aqueous phase under continuous stirring at 50°C water bath, keep warm and stir to obtain a second colostrum.

[0160] 4. The second colostrum was homogenized using a homogenizer, the homogenization pressure was controlled to be 15000 psi, the number of cycles was 4 times, the discharge temperature was 15°C, and a yellow transparent ternary supramolecular alcohol body was obtained.

[0161] Examples 13-19

[0162] These embodiments provide a ternary supramolecular alcohol body, and the specific raw material ratios are shown in Table 3 below. The ternary supramolecular body is provided in Example 1, and the amount of water used is "balance", which means that the mass of water is used to make up the total amount of the formula 100g.

[0163] Table 3

[0164]

[0165] Comparative Example 1

[0166] This comparative example provides an olive leaf extract solution, which is obtained by dissolving 10.0 g of olive leaf extract (oleuropein content ≥ 90%) in 90.0 g of deionized water.

[0167] Comparative Example 2

[0168] This comparative example provides an olive leaf extract ethosome, and the preparation method is as follows:

[0169] 1. Take 10.0 g of olive leaf extract (oleuropein content ≥ 90%), dissolve it in 45° C. water (13 g), and cool to room temperature after complete dissolution to obtain a light yellow transparent liquid, which is an oleuropein aqueous solution.

[0170] 2. Dissolve 8.0 g of lecithin (phosphatidylcholine content ≥ 90%) in 20.0 g of 1,3-propylene glycol at 60°C to obtain a transparent liquid, which is the lecithin solution.

[0171] 3. At 50° C. and under continuous stirring, slowly inject the oleuropein aqueous solution into the lecithin solution obtained in step 3 to obtain the first colostrum.

[0172] 4. Dissolve 35.0g of glycerol in 14g of water in a clean container under stirring at 50°C water bath, dissolve evenly to obtain the external aqueous phase. Then, inject the first colostrum into the external aqueous phase under 50°C water bath and continuous stirring, keep warm and stir evenly to obtain the second colostrum.

[0173] 5. The second colostrum was homogenized using a homogenizer, the homogenization pressure was controlled to be 15000 psi, the number of cycles was 4 times, and the discharge temperature was 15° C. to obtain olive leaf extract alcohol body.

[0174] Test Example 1

[0175] The products of Examples 9-19 were tested as follows.

[0176] Particle size test: The product was dispersed with laboratory pure water to obtain a 1% pure water dispersion, and then the particle size and PDI were tested using a BeNano180zeta Pro particle size tester. The test conditions were: 25°C, equilibration time 120s, and test angle 173. The obtained data are shown in Table 4.

[0177] Stability test: The product was placed in a sample bottle and placed at room temperature, -20°C, 45°C, and 4°C for 30 days. The appearance of the product was observed visually at room temperature and without direct sunlight. The data obtained are shown in Table 4.

[0178] Retention volume test:

[0179] (1) Sample preparation: Take the products of the examples, add them to the essence base respectively, stir evenly, and obtain the samples to be tested, and ensure that the content of olive leaf extract in each group of samples to be tested is consistent, wherein the content of olive leaf extract is 0.5%. All samples to be tested are prepared using a unified essence gel base, and its formula is: 3% propylene glycol, 3% glycerin, 0.8% thickener SIMULGEL FL (hydroxyethyl acrylate & sodium dimethyl taurate copolymer & isohexadecane & polysorbate 60), and the rest is water.

[0180] (2) Testing process: According to the European OECD Chemical Testing Guidelines

[0181] The in vitro transdermal test guidelines stipulated by OECD / OCDE were used for transdermal verification. The specific operation method was as follows: during the experiment, the Strat-M artificial membrane was first placed in a phosphate buffer solution (pH 7.4) for 30 minutes. After hydration, the surface moisture of the artificial membrane was wiped dry and the artificial membrane was placed between the receiving pool and the supply pool of the Franz diffusion cell, with the smooth side facing the supply pool; the receiving pool was a phosphate buffer solution (pH 7.4); 0.1g of sample was added to the supply pool; the assembled Franz diffusion cell was placed in the transdermal diffusion instrument, and the experimental conditions were set as follows: temperature was 32°C, stirring speed was 600rpm, and penetration time was 12h. The receiving solution after penetration of each group of samples was taken, and the skin retention of oleuropein was quantitatively analyzed by high performance liquid chromatography. The specific data are shown in Table 4.

[0182] Table 4

[0183]

[0184] Note: PDI refers to the aggregation degree index.

[0185] Examples 1-4 show the effects of different ratios of betaine and xylitol in binary supramolecular on the performance and stability of ternary supramolecular ethosomes. Too high or too low ratios of betaine and xylitol are not conducive to the formation of binary supramolecular liquids, and binary supramolecular liquids at room temperature may not be prepared, which may result in the inability to dissolve the olive leaf extract at room temperature or under slightly heated conditions, and the olive leaf extract may degrade and discolor when dissolved at high temperature.

[0186] Examples 5-8 show the effects of different ratios of olive leaf extract and binary supramolecule on the performance and stability of ternary supramolecule alcohol body. When the olive leaf extract is too low, the ternary supramolecule is not stable enough, mainly because the stability between the ternary supramolecules is achieved through hydrogen bonding and π-π stacking. Oleuropein provides the main source of π-π stacking, so its content plays a very important role in the stability of the ternary supramolecule. At the same time, due to the effects of hydrogen bonding and π-π stacking, the electron cloud is rearranged, and the unstable groups in oleuropein are protected, thereby achieving improved stability. When the olive leaf extract content is too high, it is difficult to dissolve at room temperature to obtain a liquid ternary supramolecule, and thus subsequent process operations cannot be performed.

[0187] Examples 13-16 show the effects of different ratios of ternary supramolecular and phospholipid on the performance and stability of ternary supramolecular alcohol bodies. If the phospholipid content is too high, some tiny empty vesicles will be formed, resulting in a large polydispersity index (PDI) of the system, which will accelerate the Ostwald ripening of the nanosystem, thereby causing particle aggregation under high temperature conditions and ultimately causing instability; and if the phospholipid content is too low, the overall particle size of the system will be large, and large liposomes will easily leak, which will also cause the high stability of the system to decrease, and its permeability will also be further reduced.

[0188] Examples 17-19 show the effects of different ratios of phospholipids and 1,3-propylene glycol on the performance and stability of the ternary supramolecular alcohol body. If the 1,3-propylene glycol content is too low, the deformability of the alcohol body membrane will decrease, thereby reducing its permeability; if the 1,3-propylene glycol content is too high, excessive short-chain alcohols will be embedded in the phospholipid bilayer, resulting in an unstable bilayer structure, and ultimately increasing the instability of the system.

[0189] Figure 1 This is the particle size test result of the ternary supramolecular alcohol body of Example 9. It can be seen that the particle size of the ternary supramolecular alcohol body prepared in Example 9 is 38.5 nm and the PDI is 0.147, indicating that the product particle size is small and uniform, which is conducive to ensuring the stability and permeability of the product.

[0190] Test Example 2

[0191] The products of Example 9, Comparative Example 1 and Comparative Example 2 were tested according to the retention test in Test Example 1. The difference from Test Example 1 was that the contents of olive leaf extract were 0.1%, 0.2% and 0.5%, respectively. The remaining sample preparation and test procedures were the same as those in Test Example 1. The obtained data were plotted to obtain Figure 2 .

[0192] from Figure 2 It can be seen that the retention amount of Comparative Example 2 is improved to a certain extent compared with Comparative Example 1, but the improvement effect is not significant; while the retention amount of oleuropein in Example 9 is significantly improved compared with Comparative Examples 1 and 2. When the concentration of olive leaf extract is 0.1%, the skin retention amount of oleuropein in Example 9 is 4.45 times that of Comparative Example 1; when the concentration of olive leaf extract is 0.2%, the skin retention amount of oleuropein in Example 9 is 3.84 times that of Comparative Example 1; when the concentration of olive leaf extract is 0.5%, the skin retention amount of oleuropein in Example 9 is 3.07 times that of Comparative Example 1. It shows that the penetration effect of the ternary supramolecular alcohol body prepared using the ternary supramolecular technology is significantly better than that of the conventional liposomes prepared without using the technology, and the formation of ternary supramolecular is very important in this scheme.

[0193] Test Example 3

[0194] The products of Example 9, Comparative Example 1 and Comparative Example 2 were placed under four conditions of room temperature, -20°C, 45°C and 4°C for 30 days, and the changes in particle size, PDI, appearance and oleuropein content were detected. The obtained data are shown in Table 5.

[0195] Table 5

[0196]

[0197]

[0198] The appearance changes of Example 9, Comparative Example 1 and Comparative Example 2 correspond to the following: Figure 3 , Figure 4 and Figure 5 shown.

[0199] Combining from Table 5 Figure 3 , Figure 4 and Figure 5 It can be seen that the olive leaf extract of Comparative Example 1, which has not undergone any treatment, deteriorates rapidly during storage, and the content decreases significantly. The high temperature has a great impact on the decrease in its content. The color of the product significantly deepens after deterioration, affecting the product quality. After the encapsulation of the conventional liposomes in Comparative Example 2, its stability is improved to a certain extent, but the high temperature still shows obvious discoloration and content decrease, indicating that oleuropein is still significantly degraded, and the quality of the product cannot be guaranteed during the normal shelf life. The ternary supramolecular alcohol body prepared in Example 9 does not show significant color change under various conditions, wherein the content of oleuropein remains stable, and the high temperature particle size increases to a certain extent, but the PDI is still less than 0.3, indicating that the particle uniformity is still very good, which helps to ensure the quality stability of the product during the normal shelf life.

[0200] Characterization Example 1

[0201] The binary supramolecular solution obtained in Example 1 was subjected to infrared spectroscopy, and the obtained spectrum was as follows: Figure 6 As shown, from Figure 6 It can be seen that after the formation of the binary supramolecular system, the apparent melting point drops, and the solid is converted into a liquid. In the infrared spectrum, the broadening of the hydroxyl C-OH stretching vibration can be observed in the wave number range of 1000-1100, and a wide and large absorption band appears at 3000-3600, which proves the formation of hydrogen bonds between hydroxyl and carboxyl groups. At the same time, the absorption of CH, CH2 and CH3 red-shifts to 2882-2932, and is also accompanied by the broadening of the absorption band; and the carboxylic acid dimer characteristics in betaine disappear, all of which confirm the formation of binary supramolecular.

[0202] The ternary supramolecule obtained in Example 1 was subjected to infrared spectrum analysis, and the spectrum obtained was as follows: Figure 7As shown, from Figure 7 It can be seen that the absorption at 1695 in the figure disappears after dissolving in the binary supramolecular, which is similar to the disappearance of the carboxyl peak of betaine during the preparation of the binary supramolecular. At the same time, the sugar structure in oleuropein is similar to the multi-hydroxy structure of xylitol, and new absorption appears at 3290 during the fusion process, which means that oleuropein has been incorporated into the original hydrogen bond system to form a ternary supramolecular system. The ternary supramolecular system of oleuropein-betaine-xylitol is a non-covalent supramolecular system that does not involve the formation or breaking of covalent bonds. Therefore, it can ensure the stability of oleuropein without affecting its overall efficacy.

[0203] The ternary supramolecule obtained in Example 1 was subjected to mass spectrometry, and the obtained spectrum is as follows Figure 8 As shown, from Figure 8 It can be seen that the ternary supramolecular system of oleuropein-betaine-xylitol is formed through hydrogen bond interaction, and its mass spectrometry is separated after ionization, indicating that no new chemical bonds are generated and the active efficacy of oleuropein is fully retained.

[0204] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A ternary supramolecule, characterized in that: Includes betaine, xylitol, and olive leaf extract; The betaine and the xylitol constitute a binary supramolecular solution, and the olive leaf extract is dissolved in the binary supramolecular solution; In the binary supramolecular solution, the molar ratio of betaine to xylitol is 1:1 to 1:3; In the ternary supramolecule, the mass ratio of the binary supramolecule solution to the olive leaf extract is 1:1-2.2; The content of oleuropein in the olive leaf extract is greater than 80%; The preparation method of the ternary supramolecule comprises the following steps: A. Add betaine and xylitol into a container, mix them evenly at 120°C to 150°C to obtain a clear liquid, and continue to cool to room temperature to obtain a binary supramolecular solution; B. dissolving the olive leaf extract in the binary supramolecular solution to obtain the ternary supramolecular solution.

2. The ternary supramolecule according to claim 1, characterized in that The content of oleuropein in the olive leaf extract is ≥85%.

3. The ternary supramolecule according to claim 1, characterized in that The content of oleuropein in the olive leaf extract is ≥90%.

4. The ternary supramolecule according to claim 1, characterized in that In step A, the mixing is performed under vacuum conditions.

5. The ternary supramolecule according to claim 4, characterized in that: The vacuum degree of the vacuum condition is ≤-0.05MPa.

6. The ternary supramolecule according to claim 1, characterized in that: In step A, the mixing time is 3h~10h.

7. The ternary supramolecule according to claim 1, characterized in that In step B, the dissolving temperature is 30°C to 50°C.

8. A ternary supramolecular alcohol body, characterized in that: According to the percentage by mass, it includes 12.5% ​​to 26.5% of ternary supramolecules, 4.0% to 10.0% of lecithin, 30.0% to 40.0% of glycerol, 15.0% to 25.0% of alcohol, and the balance is deionized water; The ternary supramolecule is the ternary supramolecule according to any one of claims 1 to 7.

9. The ternary supramolecular alcohol body according to claim 8, characterized in that: The lecithin is soybean lecithin.

10. The ternary supramolecular alcohol body according to claim 9, characterized in that: The soybean lecithin has a phosphatidylcholine content of ≥80%.

11. The ternary supramolecular alcohol body according to claim 9, characterized in that: The soybean lecithin has a phosphatidylcholine content of ≥90%.

12. The ternary supramolecular alcohol body according to claim 8, characterized in that: The alcohol is a small molecule alcohol.

13. The ternary supramolecular alcohol body according to claim 12, characterized in that: The carbon number of the small molecule alcohol is ≤5.

14. The ternary supramolecular alcohol body according to claim 12, characterized in that: The small molecule alcohol is 1,3-propylene glycol.

15. The ternary supramolecular alcohol according to any one of claims 8 to 14, characterized in that: The particle size is 20nm~100nm.

16. The ternary supramolecular alcohol according to any one of claims 8 to 14, characterized in that: The particle size is 20nm~50nm.

17. A method for preparing the ternary supramolecular alcohol body according to any one of claims 8 to 16, characterized in that: The following steps are involved: a, first dissolving the lecithin in the alcohol to obtain a lecithin solution; b. adding the ternary supramolecule into the lecithin solution to obtain the first colostrum; c. uniformly mixing the glycerol and the deionized water to obtain an external water phase; d. adding the first colostrum to the external aqueous phase and mixing them evenly to obtain a second colostrum; e. Homogenize the second colostrum to obtain the ternary supramolecular ethosome.

18. The preparation method according to claim 17, characterized in that: In step a, the dissolving temperature is 50°C to 70°C.

19. The preparation method according to claim 17, characterized in that: In step b, the temperature of the lecithin solution is 40°C to 60°C.

20. The preparation method according to claim 17, characterized in that: In step c, the mixing temperature is 40°C to 60°C.

21. The preparation method according to claim 17, characterized in that: In step d, the temperature of the external aqueous phase is 40°C to 60°C.

22. The preparation method according to claim 17, characterized in that: In step e, the homogenization pressure is 10000psi~20000psi.

23. The preparation method according to claim 17, characterized in that: The number of cycles of the homogenization is 2 to 6 times.

24. The preparation method according to claim 17, characterized in that: The homogenized discharge temperature is 8°C to 25°C.

25. Application of a ternary supramolecular alcohol body in external skin care products, characterized in that: The ternary supramolecular alcohol body is the ternary supramolecular alcohol body according to any one of claims 8 to 16; or, The ternary supramolecular alcohol body is prepared by the preparation method according to any one of claims 17 to 24.

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