EGCG supramolecule with antioxidant, liver protection and fat reduction effects, preparation method and composition thereof

Abstract

This invention discloses an EGCG supramolecular compound with antioxidant, liver-protective, and fat-reducing effects, its preparation method, and composition. The supramolecular compound is formed by non-covalent bonding between epigallocatechin gallate (EGCG) and ligand molecules containing trimethylamine and carboxyl groups; the supramolecular compound has a co-amorphous structure; the ligand molecules containing trimethylamine and carboxyl groups are betaine, ergothioneine, or L-carnitine. This invention can solve the problems of low stability and bioavailability of EGCG. It can significantly improve the solubility of EGCG and its ligands, including betaine, L-carnitine, and ergothioneine, reduce the hygroscopicity of EGCG and its ligands, including betaine, L-carnitine, and ergothioneine, and enhance the stability and bioavailability of EGCG and its ligands, including betaine, L-carnitine, and ergothioneine.

Description

Technical Field

[0001] This invention relates to the fields of medicine, food, and daily chemicals, and in particular to an EGCG supramolecular material with antioxidant, liver-protecting, and fat-reducing effects, its preparation method, and composition. Background Technology

[0002] Tea polyphenols possess various physiological activities, including antioxidant, anti-radiation, anti-aging, lipid-lowering, blood sugar-lowering, antibacterial, and enzyme-inhibiting effects. They are mainly classified into six categories: flavanones, anthocyanins, flavonols, leucosterols, and phenolic acids and condensed phenolic acids. Among these, flavanones (catechin compounds) are the most important, accounting for 60%–80% of the total tea polyphenols. Epigallocatechin gallate (EGCG) exhibits the strongest antioxidant properties, inhibiting hyaluronidase activity, reducing skin moisture loss, effectively scavenging free radicals, inhibiting photoaging, and promoting fat reduction. However, EGCG's unique chemical structure makes it prone to oxidation and discoloration, and its low bioavailability limits its applications. Therefore, a novel molecular raw material is urgently needed to address the low bioavailability of EGCG.

[0003] Supramolecular structures are molecular aggregates with specific structures and functions formed by two or more molecules (or ions) bonded together through non-covalent interactions (rather than covalent bonds). Their core is the weak intermolecular interaction rather than the strong intramolecular covalent bond. Examples include cyclodextrin inclusion systems, supramolecular gel drug delivery systems, and co-crystal / co-amorphous supramolecular systems. Co-amorphous formulations involve preparing two or more active ingredients into a homogeneous solid dispersion without a fixed crystalline structure. Co-amorphous formulations can improve stability, reduce activity loss, enhance bioavailability, and improve absorption efficiency.

[0004] Currently, there are few applications of preparing EGCG as supramolecular components. CN119185589A discloses a self-assembled supramolecular raw material of hydroxypropyl-β-cyclodextrin and epigallocatechin gallate betaine, its preparation method, and its application. The supramolecular component uses epigallocatechin gallate betaine cocrystals. However, there are currently no reports on the preparation of EGCG and ligands into co-amorphous forms. Summary of the Invention

[0005] To address the aforementioned problems in existing technologies, this invention proposes an EGCG supramolecular compound with antioxidant, liver-protective, and fat-reducing effects, its preparation method, and composition. This solves the problems of low stability and bioavailability of epigallocatechin gallate (EGCG). It can significantly improve the solubility of EGCG and its ligands, including betaine, L-carnitine, and ergothioneine, reduce the hygroscopicity of EGCG or its ligands, including betaine, L-carnitine, and ergothioneine, and enhance the stability and bioavailability of EGCG and its ligands, including betaine, L-carnitine, and ergothioneine.

[0006] Furthermore, EGCG, when combined with its ligands betaine and L-carnitine to form supramolecular structures, can synergistically enhance its fat-reducing effects. EGCG, when combined with its ligand ergothioneine to form supramolecular structures, can synergistically enhance its antioxidant and anti-aging effects.

[0007] The technical solution of the present invention is as follows: The first objective of this invention is to provide an EGCG supramolecular material with antioxidant, liver-protecting and fat-reducing effects, which is formed by non-covalent bonding of epigallocatechin gallate (EGCG) and ligands containing trimethylamine and carboxyl groups; the supramolecular material has a co-amorphous structure.

[0008] In one embodiment of the present invention, the ligand containing trimethylamine and carboxyl groups is betaine, ergothioneine, or L-carnitine.

[0009] In one embodiment of the present invention, the molar ratio of EGCG to a ligand containing trimethylamine and carboxyl groups is 1:1-2.

[0010] In one embodiment of the present invention, the molar ratio of EGCG to betaine is 1:2.

[0011] In one embodiment of the present invention, the molar ratio of EGCG to L-carnitine is 1:1.

[0012] In one embodiment of the present invention, the molar ratio of EGCG to ergothioneine is 1:1.

[0013] In one embodiment of the present invention, the melting point of EGCG is 219°C-220°C, the melting point of supramolecular epigallocatechin gallate-betaine is 196-205°C, the melting point of supramolecular epigallocatechin gallate-L-carnitine is 168-187°C, and the melting point of supramolecular epigallocatechin gallate-ergothioneine is 172-176°C.

[0014] The second objective of this invention is to provide a method for preparing the above-mentioned EGCG supramolecular material with antioxidant, liver-protective, and fat-reducing effects, comprising the following steps: EGCG was dissolved in water with a ligand containing trimethylamine and carboxyl groups, and then freeze-dried to obtain the EGCG supramolecular having antioxidant, liver-protective and fat-reducing effects.

[0015] In one embodiment of the present invention, the ratio of the total mass of EGCG and the ligand containing trimethylamine and carboxyl groups to water is 1:2-20 g / mL; the freeze-drying conditions are: pre-freezing at -80℃ for 12-20 h, then transferring to a freeze dryer and drying at -30℃ and 5-10 Pa for 24-30 h, and then drying at 20℃ and 5-8 Pa for 8 h.

[0016] In one embodiment of the present invention, a method for preparing EGCG supramolecular molecules with antioxidant, liver-protective, and fat-reducing effects includes the following steps: S1. Add EGCG, ligands containing trimethylamine and carboxyl groups, and water to a reaction vessel, and stir to fully mix the raw materials. The ratio of the total mass of EGCG and ligands containing trimethylamine and carboxyl groups to water is 1:2-20 g / mL, preferably 1:2-4 g / mL. The mixing conditions are: 15-25℃, 0.5-5h, preferably 1-3h. S2. Freeze-dry the solution obtained in S1. The freeze-drying conditions are as follows: pre-freeze at -80℃ for 12-20 h, then transfer to a freeze dryer and dry at -30℃ and 5-10 Pa for 24-30 h, and then dry at 20℃ and 5-8 Pa for 8 h.

[0017] The third objective of this invention is to provide a method for preparing the above-mentioned EGCG supramolecular material with antioxidant, liver-protective, and fat-reducing effects, comprising the following steps: EGCG is pulverized and mixed with ligands containing trimethylamine and carboxyl groups, heated to a molten state, then immersed in a cooling medium and cooled to complete solidification. After cooling to room temperature, it is pulverized and sieved to obtain the EGCG supramolecular with antioxidant, liver-protecting and fat-reducing effects.

[0018] In one embodiment of the present invention, the temperature is heated to 180-200°C; the cooling medium is dry ice or liquid nitrogen; and the sieve mesh size is 80-100 mesh.

[0019] In one embodiment of the present invention, EGCG is pulverized and mixed with a ligand containing trimethylamine and carboxyl groups, heated to a molten state, and then rapidly immersed in liquid nitrogen for quenching.

[0020] In one embodiment of the present invention, a method for preparing EGCG supramolecular molecules with antioxidant, liver-protective, and fat-reducing effects includes the following steps: S1, EGCG and a ligand containing trimethylamine and carboxyl groups are pulverized into a fine powder and mixed to form a mixed powder; the ligand is betaine, L-carnitine or ergothioneine; the molar ratio of EGCG to ligand is 1:1-2; S2, place the mixed powder into a crucible, and under nitrogen protection, slowly heat it to 180-200℃ and hold it for 10-20 minutes, stirring gently until it is completely melted and forms a uniform molten mixture; S3, the molten mixture is immersed in a cooling medium to cool the material until it is completely solidified; the cooling medium is dry ice or liquid nitrogen, preferably liquid nitrogen; S4. The solid obtained in S3 is left to return to room temperature, and then the solidified block is taken out, crushed and sieved. The solidified block is then passed through an 80-100 mesh sieve to obtain supramolecular powder.

[0021] The fourth objective of this invention is to provide an application of the above-mentioned EGCG supramolecular material with antioxidant, liver-protecting and fat-reducing effects for the preparation of pharmaceuticals, food or daily chemical products.

[0022] The fifth objective of this invention is to provide a composition comprising the above-mentioned EGCG supramolecular molecules with antioxidant, liver-protecting and fat-reducing effects.

[0023] In one embodiment of the present invention, the composition further contains pharmaceutically acceptable excipients.

[0024] In one embodiment of the invention, the composition further contains an excess of EGCG and / or an excess of the corresponding ligand (betaine, L-carnitine, ergothioneine), as well as other pharmaceutically acceptable excipients.

[0025] In other words, the molar ratio of EGCG to the corresponding ligand in the raw materials of the composition is not particularly limited, as long as the raw materials of the composition can be used to prepare the above-mentioned EGCG supramolecular.

[0026] For example, the molar ratio of EGCG to betaine in the composition can be 10:1 to 1:10, wherein a portion of the component exists in the form of EGCG supramolecular molecules, while the other portion exists in a free form. Preferably, all betaine is formed into supramolecular molecules to overcome the highly hygroscopic nature of betaine. In the preparation method of the composition, the molar ratio of betaine to EGCG is 10:1 to 1:10, more preferably 2:1.01 to 2:5.

[0027] For example, the molar ratio of EGCG to L-carnitine in the composition can be 10:1 to 1:10, wherein a portion of the component exists in the form of EGCG supramolecular molecules, while the other portion exists in a free form. Preferably, all L-carnitine is formed into supramolecular molecules to overcome the highly hygroscopic nature of L-carnitine. In the preparation method of the composition, the molar ratio of L-carnitine to EGCG is 10:1 to 1:10, more preferably 1:1.01 to 1:5.

[0028] For example, the molar ratio of EGCG to ergothioneine in the composition can be 10:1 to 1:10, with one component existing in the form of EGCG supramolecular molecules and the other component existing in free form. Preferably, all EGCG is formed into supramolecular molecules to overcome the drawbacks of EGCG, such as easy degradation and discoloration during long-term storage and low bioavailability. In the preparation method of the composition, the molar ratio of EGCG to ergothioneine is 10:1 to 1:10, more preferably 1:1.01 to 1:5.

[0029] The beneficial technical effects of this invention are as follows: This invention prepares EGCG with ligands containing specific functional groups into a supramolecular co-amorphous form to improve the stability, bioavailability and efficacy of EGCG.

[0030] The supramolecular structure is made by combining EGCG with ligands that have specific functional groups (possessing both trimethylamine and carboxyl groups) using supramolecular co-amorphous technology. This supramolecular structure can not only solve the problem of EGCG being prone to degradation and discoloration during long-term storage, but also improve solubility and enhance bioavailability. This supramolecular structure is easy to mass-produce.

[0031] The supramolecular structure formed by EGCG and ligands through weak interactions such as hydrogen bonding and van der Waals interactions in this invention fully solves the problem of EGCG's easy oxidation and discoloration compared to EGCG itself.

[0032] The supramolecular structure of this invention exhibits significantly improved solubility and synergistic effects, promoting penetration, significantly enhancing bioavailability, and improving absorption efficiency. It also significantly reduces the irritation of EGCG, avoiding the risk of damage to the skin and intestines, achieving gentle and efficient drug delivery, and laying the foundation for its widespread application in pharmaceuticals, food, and daily chemicals.

[0033] Compared to EGCG or ligands, the supramolecular structure of this invention significantly reduces hygroscopicity, effectively addresses the problems of ligands being prone to moisture absorption, clumping, and deliquescence, and improves the physical stability and ease of use of the product during storage and application, providing reliable support for subsequent industrial production and application.

[0034] The supramolecular epigallocatechin gallate-betaine and supramolecular epigallocatechin gallate-L-carnitine of the present invention have significantly improved fat reduction and weight loss capabilities compared to EGCG, betaine, and L-carnitine themselves.

[0035] The supramolecular epigallocatechin gallate-ergothioneine of this invention has significantly improved antioxidant and anti-aging capabilities compared to EGCG and ergothioneine itself. Attached Figure Description

[0036] Figure 1 The structural formula of supramolecular epigallocatechin gallate-betaine is given.

[0037] Figure 2 The structural formula is that of supramolecular epigallocatechin gallate-L-carnitine.

[0038] Figure 3 The structural formula is that of supramolecular epigallocatechin gallate-ergothionein.

[0039] Figure 4 X-ray diffraction (PXRD) patterns of EGCG, betaine, and supramolecular epigallocatechin gallate-betaine powder obtained in Example 1.

[0040] Figure 5 X-ray diffraction (PXRD) patterns of EGCG, L-carnitine, and the supramolecular epigallocatechin gallate-L-carnitine powder obtained in Example 2.

[0041] Figure 6 X-ray diffraction (PXRD) patterns of EGCG, ergothioneine, and the supramolecular epigallocatechin gallate-ergothioneine powder obtained in Example 3.

[0042] Figure 7 This is a photograph of the supramolecular epigallocatechin gallate-betaine obtained in Example 1.

[0043] Figure 8 This is a photograph of the supramolecular epigallocatechin gallate-L-carnitine obtained in Example 2.

[0044] Figure 9 This is a photograph of the supramolecular epigallocatechin gallate-ergothioneine obtained in Example 3.

[0045] Figure 10 The solubility results of EGCG, betaine, and supramolecular epigallocatechin gallate-betaine obtained in Example 1 are shown in the figure.

[0046] Figure 11 The figure shows the solubility results of EGCG, L-carnitine, and the supramolecular epigallocatechin gallate-L-carnitine obtained in Example 2.

[0047] Figure 12 The figure shows the solubility results of EGCG, ergothioneine, and the supramolecular epigallocatechin gallate-ergothioneine obtained in Example 3.

[0048] Figure 13 Figure showing the results of EGCG, betaine, L-carnitine, supramolecular epigallocatechin gallate-betaine prepared in Example 1, and supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2 inhibiting adipogenic differentiation of mouse embryonic fibroblasts.

[0049] Figure 14 Statistical graph showing the inhibition of adipogenic differentiation of mouse embryonic fibroblasts by EGCG, betaine, L-carnitine, supramolecular epigallocatechin gallate-betaine prepared in Example 1, and supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2.

[0050] Figure 15 Results of oral bioavailability of supramolecular epigallocatechin gallate-betaine.

[0051] Figure 16 Results of oral bioavailability of supramolecular epigallocatechin gallate-L-carnitine.

[0052] Figure 17 Results of oral bioavailability of supramolecular epigallocatechin gallate-ergothioneine.

[0053] Figure 18 The results show the changes in rat body weight.

[0054] Figure 19 The results show the visceral fat coefficient of rats.

[0055] Figure 20 The results show the triglyceride (TG) levels in rat blood lipids.

[0056] Figure 21 The results show the total cholesterol (TC) levels in rat blood lipids.

[0057] Figure 22 The results show the HDL-C index in rat blood lipids.

[0058] Figure 23 The results show the LDL-C level in rat blood lipids.

[0059] Figure 24 Statistical results on the anti-photoaging effects of EGCG, ergothioneine, and supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3.

[0060] Figure 25 Figure showing the anti-free radical aging results of EGCG, ergothioneine, and supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3.

[0061] Figure 26 Statistical graph of the anti-free radical aging results of EGCG, ergothioneine, and supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3.

[0062] Figure 27 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-betaine powder obtained in Example 4.

[0063] Figure 28 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-L-carnitine powder obtained in Example 5.

[0064] Figure 29 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-ergothioneine powder obtained in Example 6.

[0065] Figure 30 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-betaine composition powder obtained in Example 7.

[0066] Figure 31 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-L-carnitine composition powder obtained in Example 8.

[0067] Figure 32 The image shows the X-ray diffraction (PXRD) pattern of the supramolecular epigallocatechin gallate-ergothioneine composition powder obtained in Example 9. Detailed Implementation

[0068] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing specific embodiments only and is not intended to limit the present invention.

[0069] Reagents and Instruments X-ray powder diffractometer (D8 Discover with TXS, Bruker. AXS), differential scanning calorimeter (Mettler Toledo), Fourier transform infrared spectrometer (PerkinEelmer), freeze dryer (YAMATO), high performance liquid chromatograph (HPLC, Agilent), ultrasonic reactor (Ymnl-5LF, Nanjing Emmanuel Instruments), ergothioneine (Shenzhen Zhongke Xinyang Biotechnology Co., Ltd.), L-tyrosine (Maclean), L-proline (Maclean), L-carnitine (Maclean), betaine (Maclean), epigallocatechin gallate (Xihai Biotechnology), ultra-high performance liquid chromatography-tandem mass spectrometry.

[0070] Example 1 A method for preparing supramolecular epigallocatechin gallate-betaine includes the following steps: S1. Add 1 mol EGCG, 2 mol betaine, and 2.070 L of sterile water to the reactor, stir at 800 rpm, and react at 20°C for 2 h. S2. The solution obtained in S1 is pre-frozen at -80℃ for 20 h, and then transferred to a freeze dryer and dried at -30℃ and 10 Pa for 24 h to obtain supramolecular epigallocatechin gallate-betaine powder.

[0071] Figure 1 The structural formula of supramolecular epigallocatechin gallate-betaine; Figure 4 The image shows the X-ray diffraction patterns of EGCG, betaine, and the supramolecular epigallocatechin gallate-betaine powder obtained in this example. The pattern illustrates the relationship between intensity (I; count) and angle 2θ (°). EGCG and its ligand betaine both exhibit characteristic diffraction peaks at 2θ. The supramolecular epigallocatechin gallate-betaine powder shows only a large, diffuse diffraction ring, and the characteristic peaks of EGCG and its ligand disappear, indicating that the product has formed a supramolecular co-amorphous state. Figure 7 Supramolecular epigallocatechin gallate-betaine powder is a pink powder.

[0072] Example 2 A method for preparing supramolecular epigallocatechin gallate-L-carnitine includes the following steps: S1. Add 1 mol EGCG, 1 mol L-carnitine, and 1.238 L of sterile water to the reactor, stir at 1000 rpm, and react at 22℃ for 2.5 h. S2. The solution obtained in S1 is pre-frozen at -80℃ for 12 h, and then transferred to a freeze dryer and dried at -30℃ and 10 Pa for 24 h to obtain supramolecular epigallocatechin gallate-L-carnitine powder.

[0073] Figure 2 The structural formula of supramolecular epigallocatechin gallate-L-carnitine; Figure 5 The image shows the X-ray diffraction patterns of EGCG, L-carnitine, and the supramolecular epigallocatechin gallate-L-carnitine powder obtained in this example. The pattern illustrates the relationship between intensity (I; count) and angle 2θ (°). EGCG and its ligand L-carnitine both exhibit characteristic diffraction peaks at 2θ. Only one large diffraction ring was observed in the supramolecular epigallocatechin gallate-L-carnitine, which appears diffuse. The characteristic peaks of EGCG and its ligand disappear, indicating that the product has formed a supramolecular co-amorphous state. Figure 8 Supramolecular epigallocatechin gallate-L-carnitine powder is a pink powder.

[0074] Example 3 A method for preparing supramolecular epigallocatechin gallate-ergothioneine includes the following steps: S1. Add 1 mol EGCG, 1 mol ergothioneine, and 2.061 L of sterile water to the reactor, stir at 1300 rpm, and react at 22°C for 1.5 h. S2. The solution obtained in S1 is pre-frozen at -80℃ for 12 h, and then transferred to a freeze dryer and dried at -30℃ and 10 Pa for 30 h to obtain supramolecular epigallocatechin gallate-ergothioneine powder.

[0075] Figure 3 The structural formula of supramolecular epigallocatechin gallate-ergothioneine is given. Figure 6 The X-ray diffraction patterns of EGCG, ergothioneine, and the supramolecular epigallocatechin gallate-ergothioneine powder obtained in this example are shown. The pattern illustrates the relationship between intensity (I; count) and angle 2θ (°). EGCG and its ligand ergothioneine both exhibit characteristic diffraction peaks at the 2θ value. Only one large diffraction ring was observed in the supramolecular epigallocatechin gallate-ergothioneine, and the X-ray powder diffraction pattern was diffuse. The characteristic peaks of EGCG and its ligand disappeared, proving that the product formed a supramolecular co-amorphous state.

[0076] Example 4 Supramolecular epigallocatechin gallate-betaine was prepared by melt-quenching method. S1. Add 1 mol EGCG and 2 mol betaine to a grinder and grind them into fine powder and mix evenly.

[0077] S2. Place the mixed powder into a crucible, slowly heat it to 200°C under nitrogen protection and hold it at that temperature for 10-15 minutes, stirring gently until it is completely melted and forms a uniform molten mixture.

[0078] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0079] S4. Place the solid obtained in S3 at room temperature until it returns to normal temperature, then take out the solidified block, crush and sieve it, and pass it through an 80-mesh sieve to obtain supramolecular powder.

[0080] PXRD test results are as follows Figure 27 As shown, similar to Example 1, the spectrum is diffuse, proving that the product has formed a supramolecular co-amorphous state.

[0081] Example 5 Supramolecular epigallocatechin gallate-L-carnitine was prepared using a melt-quenching method. S1. Add 1 mol EGCG and 1 mol L-carnitine to a grinder and grind them into fine powder and mix them evenly.

[0082] S2. Place the mixed powder into a crucible, slowly heat it to 190°C under nitrogen protection and hold it at that temperature for 15-20 minutes, stirring gently until it is completely melted and forms a uniform molten mixture.

[0083] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0084] S4. Place the solid obtained in S3 at room temperature until it returns to normal temperature, then take out the solidified block, crush and sieve it, and pass it through an 80-mesh sieve to obtain supramolecular powder.

[0085] PXRD test results are as follows Figure 28 As shown, similar to Example 2, the spectrum is diffuse, proving that the product has formed a supramolecular co-amorphous state.

[0086] Example 6 Supramolecular epigallocatechin gallate-ergothioneine was prepared using a melt-quenching method. S1. Add 1 mol EGCG and 1 mol ergothioneine to a grinder and grind them into fine powder and mix them evenly.

[0087] S2. Place the mixed powder into a crucible, slowly heat it to 180°C under nitrogen protection and hold it for 10-15 minutes, stirring gently until it is completely melted and forms a uniform molten mixture.

[0088] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0089] S4. Place the solid obtained in S3 at room temperature until it returns to normal temperature, then take out the solidified block, crush and sieve it, and pass it through an 80-mesh sieve to obtain supramolecular powder.

[0090] PXRD test results are as follows Figure 29 As shown, similar to Example 3, the spectrum is diffuse, proving that the product has formed a supramolecular co-amorphous state.

[0091] Example 7 Supramolecular epigallocatechin gallate-betaine composition was prepared by melt-quenching method. S1. Add 5 mol EGCG and 1 mol betaine to a grinder and grind them into fine powder and mix evenly.

[0092] S2. Place the mixed powder into a crucible, slowly heat it to 200℃ under nitrogen protection and keep it at that temperature for 10-15 minutes, stirring gently until the betaine is completely melted and forms a uniform molten mixture.

[0093] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0094] S4. The solid obtained in S3 is left to stand at room temperature until it returns to normal temperature. The solidified block is then removed, crushed, and sieved through an 80-mesh sieve to obtain a supramolecular epigallocatechin gallate-betaine composition powder. Figure 30 As shown, PXRD analysis revealed that the composition exhibits a large diffraction ring similar to that of the supramolecular epigallocatechin gallate-betaine prepared in Example 1, as well as characteristic diffraction peaks of the EGCG monomer. This confirms the presence of supramolecular epigallocatechin gallate-betaine and EGCG monomer within the composition.

[0095] Example 8 A supramolecular epigallocatechin gallate-L-carnitine composition was prepared using a melt-quenching method. S1. Add 2 mol EGCG and 1 mol L-carnitine to a grinder and grind them into fine powder and mix evenly.

[0096] S2. Place the mixed powder into a crucible, slowly heat it to 190°C under nitrogen protection and keep it at that temperature for 10-15 minutes, stirring gently until the L-carnitine is completely melted and forms a uniform molten mixture.

[0097] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0098] S4. The solid obtained in S3 is left to return to room temperature, then the solidified block is removed, crushed, and sieved through an 80-mesh sieve to obtain a supramolecular epigallocatechin gallate-L-carnitine composition powder. Figure 31 As shown, PXRD analysis revealed that the composition exhibits large diffraction rings and characteristic diffraction peaks of the EGCG monomer, similar to those of the supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2. This demonstrates the presence of supramolecular epigallocatechin gallate-L-carnitine and EGCG monomers in the composition.

[0099] Example 9 A supramolecular epigallocatechin gallate-ergothioneine composition was prepared using a melt-quenching method. S1. Add 1 mol EGCG and 2 mol ergothioneine to a grinder and grind them into fine powder and mix them evenly.

[0100] S2. Place the mixed powder into a crucible, slowly heat it to 180°C under nitrogen protection and hold it for 15-20 minutes, stirring gently until EGCG is completely melted and forms a uniform molten mixture.

[0101] S3. Immerse the molten mixture obtained in S2 into liquid nitrogen and cool the material until it is completely solidified.

[0102] S4. The solid obtained in S3 is left to stand at room temperature until it returns to normal temperature. The solidified block is then removed, crushed, and sieved through an 80-mesh sieve to obtain a supramolecular epigallocatechin gallate-ergothioneine composition powder. Figure 32 As shown, PXRD analysis revealed that the composition exhibits a large diffraction ring and characteristic diffraction peaks of the ergothioneine monomer, similar to those of the supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3. This demonstrates the presence of supramolecular epigallocatechin gallate-ergothioneine and ergothioneine monomers within the composition.

[0103] Comparative Example 1 0.5 mol of L-proline and 0.5 mol of EGCG were added to 5 L of water, stirred for 4 h, and freeze-dried to obtain a solid powder. PXRD analysis showed that the X-ray diffraction peaks of the powder were those of L-proline or EGCG, with no new or decreased characteristic peaks, proving that the powder was free L-proline and EGCG, and supramolecular L-proline-epigallocatechin gallate was not obtained.

[0104] Comparative Example 2 0.5 mol of L-tyrosine and 0.5 mol of EGCG were added to 5 L of water, stirred for 4 h, and freeze-dried to obtain a solid powder. PXRD analysis showed that the X-ray diffraction peaks of the powder were those of L-tyrosine or EGCG, with no new or decreased characteristic peaks, proving that the powder was free L-tyrosine and free EGCG, and supramolecular L-tyrosine-epigallocatechin gallate was not obtained.

[0105] Comparative Example 3 1 mol of EGCG and 1 mol of L-carnitine were added to a grinder and pulverized into a fine powder, then mixed evenly. The powder mixture was placed in a crucible and slowly heated to 230°C under nitrogen protection, holding the temperature for 15-20 minutes while gently stirring until completely melted and forming a homogeneous molten mixture. The molten mixture was then immersed in liquid nitrogen and cooled until completely solidified. The resulting solid was allowed to return to room temperature, and the solidified block was then removed, pulverized, and sieved through an 80-mesh sieve to obtain powder. HPLC analysis showed that the EGCG content in the powder was only 4%, indicating that the excessively high melting temperature caused EGCG decomposition, and that this melting temperature was unsuitable for preparing high-purity supramolecular amorphous materials.

[0106] Comparative Example 4 1 mol of EGCG and 1 mol of ergothioneine were added to a pulverizer and pulverized into a fine powder, then mixed evenly. The mixed powder was placed in a crucible and slowly heated to 150°C under nitrogen protection, and held at that temperature for 20-30 minutes, stirring gently during the process. The resulting molten mixture was then immersed in liquid nitrogen and cooled until completely solidified. The resulting solid was allowed to return to room temperature, and the solidified block was then removed, pulverized, and sieved through an 80-mesh sieve to obtain powder. PXRD analysis showed that the X-ray diffraction pattern of the powder contained the characteristic diffraction peaks of EGCG and ergothioneine, with no newly added or disappeared characteristic peaks, proving that high-purity supramolecular amorphous material could not be prepared at this temperature.

[0107] Test example: (1) Solubility Take 0.01 mol of EGCG, betaine, L-carnitine, ergothioneine, supramolecular epigallocatechin gallate-betaine prepared in Example 1, supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2, and supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3, add 10 mL of sterile water, shake vigorously for 10 min, and observe whether the powder is completely dissolved.

[0108] like Figure 10-12 It was observed that 1M EGCG and ergothioneine were insoluble, while supramolecular epigallocatechin gallate-betaine, supramolecular epigallocatechin gallate-L-carnitine, and supramolecular epigallocatechin gallate-ergothioneine were all completely soluble at a concentration of 1M, proving that preparing EGCG into a supramolecular coamorphous form can enhance the solubility of EGCG.

[0109] (2) Stability EGCG, the supramolecular epigallocatechin gallate-betaine prepared in Example 1, the supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2, and the supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3 were placed under different conditions, and their contents were evaluated. The content of EGCG was determined by HPLC, and the results are shown in Table 1 below.

[0110] When storing materials in open, transparent glass bottles, the following conditions must be met: Condition 1: Temperature 40 ± 2℃, humidity 75 ± 5%, 30-180 days; Condition 2: Temperature 25 ± 2℃, humidity 65 ± 5%, 30-180 days; Table 1

[0111] As shown in Table 1, under condition 1, the content of free EGCG decreased to 43.86% within 180 days, while the supramolecular epigallocatechin gallate-betaine, epigallocatechin gallate-L-carnitine, and epigallocatechin gallate-ergothioneine prepared in Example 1 could all maintain a content of over 98.3%. Under condition 2, the content of free EGCG decreased to 69.46% within 180 days, while the supramolecular epigallocatechin gallate-betaine, epigallocatechin gallate-L-carnitine, and epigallocatechin gallate-ergothioneine prepared in Example 1 could all maintain a content of over 99%, demonstrating that preparing EGCG into supramolecular form can significantly improve its stability.

[0112] (3) Hygroscopicity Take a dry, stoppered glass weighing bottle (outer diameter 50 mm, height 15 mm) and place it in a suitable 25℃±1℃ constant temperature desiccator (with ammonium chloride or ammonium sulfate saturated solution at the bottom) or artificial climate chamber (temperature set at 25℃±1℃, relative humidity at 80%±2%) one day before the test, and accurately weigh it (m1).

[0113] Take appropriate amounts of EGCG, betaine, L-carnitine, ergothioneine, supramolecular epigallocatechin gallate-betaine prepared in Example 1, supramolecular epigallocatechin gallate-L-carnitine prepared in Example 2, and supramolecular epigallocatechin gallate-ergothioneine prepared in Example 3, and spread them evenly in the weighing bottle. The sample thickness is generally about 1 mm, and the weight (m2) is accurately measured.

[0114] Leave the weighing bottle open and place it, along with the cap, under the aforementioned constant temperature and humidity conditions for 24 hours.

[0115] Close the weighing bottle cap and accurately weigh it (m3).

[0116] Percentage of weight gain

[0117] Description of hygroscopic characteristics and definition of hygroscopic weight gain Deliquescence: The process of absorbing sufficient water to form a liquid.

[0118] Extremely hygroscopic: the weight gain due to moisture absorption is not less than 15%.

[0119] It has hygroscopic properties: the weight gain due to moisture absorption is less than 15% but not less than 2%.

[0120] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%.

[0121] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0122] Table 2

[0123] As shown in Table 2, EGCG is slightly hygroscopic, betaine is extremely hygroscopic, and the supramolecular epigallocatechin gallate-betaine is hygroscopic. L-carnitine is extremely hygroscopic, the supramolecular epigallocatechin gallate-L-carnitine is slightly hygroscopic, ergothioneine is hygroscopic or almost hygroscopic, and the supramolecular epigallocatechin gallate-ergothioneine is hygroscopic or almost hygroscopic. This demonstrates that preparing EGCG into supramolecular co-amorphous forms with its ligands betaine, L-carnitine, and ergothioneine respectively can significantly reduce hygroscopicity. In particular, supramolecular epigallocatechin gallate-betaine and supramolecular epigallocatechin gallate-L-carnitine reduce the hygroscopicity of their ligands to hygroscopicity or slight hygroscopicity, effectively changing the problems of ligands being prone to moisture absorption, clumping, and poor stability, and improving the physical stability and ease of use of the products during storage and application, providing reliable support for subsequent industrial production and application.

[0124] (4) Evaluation of the efficacy of inhibiting adipogenic differentiation of 3T3-L1 cells The supramolecular epigallocatechin gallate (EGCG), betaine (TMG), and L-carnitine (L-Carn group) prepared in Example 1, namely supramolecular epigallocatechin gallate-betaine (supramolecular EGCG-TMG), and supramolecular epigallocatechin gallate-L-carnitine (supramolecular EGCG-L-Carn) prepared in Example 2, were used to inhibit the differentiation of mouse embryonic fibroblasts (3T3-L1) cells to evaluate their ability to inhibit cell adipogenesis.

[0125] Mouse embryonic fibroblasts (3T3-L1) cells were divided into groups of 5 × 10⁻⁶. 4 Cells were seeded in 6-well plates and cultured in DMEM medium containing 10% fetal bovine serum for 48 hours until the cell density reached over 90%. The medium was then replaced with a cocktail medium containing the test substance (DMEM medium containing 0.5 mM IBMX, 1 μM MDEX, 10 μg / mL insulin, and 10% fetal bovine serum) to induce cell differentiation. The blank group was not induced and did not contain the test substance, the model group was induced and did not contain the test substance, and the other groups were induced and given the corresponding test substance at a concentration of 10 mg / L. After 48 hours of culture, the cocktail medium was replaced with maintenance medium containing the test substance (DMEM medium containing 10 μg / mL insulin and 10% fetal bovine serum). The maintenance medium was changed every two days for a total of 4 times over 8 days. After Oil Red O staining, the cells were photographed under a microscope, and the percentage of lipid droplet area was counted.

[0126] The lipid droplet distribution maps and statistical results for each group are as follows: Figure 13-14As shown, compared with the blank group, the percentage of lipid droplet area in the model group increased by 36.98%, and the difference was statistically significant. p <0.01 indicates successful modeling and valid experimental results. Compared with the model group, the percentage of lipid droplet area in the EGCG group decreased by 22.47% ( p <0.01), the TGM group decreased by 3.59% ( p >0.05%, the supramolecular EGCG-TGM group decreased by 35.40% ( p <0.01), the L-Carn group experienced a 27.29% reduction in body weight ( p <0.01%, the supramolecular EGCG-L-Carn group experienced a 38.21% reduction in body weight ( p <0.01). The percentage of lipid droplet area decreased to some extent in all treatment groups. Among them, the supramolecular EGT-EGCG group showed a more significant ability to inhibit adipogenic differentiation of 3T3-L1 cells compared with the EGCG group administered alone, the ligand group and the mixture group.

[0127] (5) Evaluation of oral bioavailability Twenty-five male SPF-grade SD rats, weighing 250–300 g, were used and divided into four groups: a control group, an epigallocatechin gallate group (EGCG group), a supramolecular epigallocatechin gallate-betaine group (supramolecular EGCG-TMG group), a supramolecular epigallocatechin gallate-L-carnitine group (supramolecular EGCG-L-Carn group), and a supramolecular epigallocatechin gallate-ergothioneine group (supramolecular EGCG-EGT group), with five rats in each group. The oral gavage dose was 100 mg / kg.

[0128] Before the experiment, rats were fasted for 12 hours. Blood was collected from the tail tip at 0, 10, 20, 40, 60, 120, 180, 240, and 360 minutes after gavage, with a blood volume of approximately 0.3-0.4 mL each time. The serum was stored at -80℃ for later use. 200 μL of the supernatant was extracted with ethyl acetate by vortexing, dried under nitrogen, dissolved completely in methanol, and filtered through a 0.22 μm filter. The EGCG content was analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry. Pharmacokinetic curves were plotted, and the time to maximum plasma EGCG concentration (T0) was determined. max ), maximum plasma EGCG concentration (C max ) and the area under the curve (AUC).

[0129] The result is as follows Figure 15-17As shown in Table 3, in the EGCG group, the plasma EGCG concentration reached its highest level (248.33±50.08 μg / L) 40 min after a single oral EGCG dose, and then rapidly decreased, with an AUC (0-360 min) of 27987±2084 μg / L·min. In contrast, the plasma EGCG concentrations of supramolecular epigallocatechin gallate-betaine reached their highest levels at 20 min (682.01±42.58 μg / L), supramolecular epigallocatechin gallate-L-carnitine at 40 min (749.33±6.35 μg / L), and supramolecular epigallocatechin gallate-ergothioneine at 20 min (749.33±6.35 μg / L), all significantly higher than those in the EGCG group, and their AUCs were also significantly lower. （0-360 min） The values ​​were 2.88 times, 2.92 times, and 2.90 times, respectively, indicating that preparing EGCG as a supramolecular co-amorphous material can improve the bioavailability of EGCG.

[0130] Table 3

[0131] (6) Evaluation of fat reduction efficacy The fat-reducing efficacy of the supramolecular epigallocatechin gallate-betaine group (supramolecular EGCG-TMG group) prepared in Example 1, the supramolecular epigallocatechin gallate-L-carnitine group (supramolecular EGCG-L-Carn group) prepared in Example 2, the epigallocatechin gallate group (EGCG group), the betaine group (TMG group), and the L-carnitine group (L-Carn group) was evaluated.

[0132] One hundred and fifteen male SPF-grade SD rats, weighing 145–155 g, were used. After 7 days of acclimatization feeding and a 12-hour fast, they were weighed, excluding rats with significant weight differences. Fifteen rats were used as the control group, fed a normal diet and purified water. The remaining rats were used as the model group, fed a high-fat diet and purified water. Eight weeks after modeling, they were weighed and blood was collected from their tails to detect lipid-related indicators (serum total cholesterol and triglycerides). The success of the model was evaluated, and rats with significant deviations in weight and lipid-related indicators between the control and model groups were excluded. Ten rats with similar weight and lipid-related indicators were selected for the control group, and 80 rats were selected for the model group. The model group was randomly divided into a model group, a supramolecular EGT-EGCG group, an EGCG group, an EGT group, and an EGCG+EGT mixture group, with ten rats in each group. The specific administration method is as follows: Administer the medication via gavage once daily for 6 weeks.

[0133] S1, Control Group: fed normal feed + purified water; S2, Model Group: Feeded with high-fat diet and purified water; S3, EGCG group: fed high-fat diet + purified water + 50mg / kg epigallocatechin gallate; S4 and TMG groups: fed with high-fat feed + purified water + 50 mg / kg betaine; S5, EGCG+TMG mixed group: fed high-fat diet + purified water + 10.17 mg / kg betaine + 39.83 mg / kg EGCG; S6, supramolecular EGCG-TMG group: fed with high-fat diet + purified water + 50mg / kg supramolecular epigallocatechin gallate-betaine; S7, L-Carn group: fed high-fat diet + purified water + 50mg / kg L-carnitine; S8, EGCG+L-Carn mixed group: fed high-fat diet + purified water + 13.00 mg / kg L-carnitine + 37.00 mg / kg EGCG; S9, supramolecular EGCG-L-Carn group: fed with high-fat diet + purified water + 50mg / kg supramolecular epigallocatechin gallate-L-carnitine; After the experiment, the rats were fasted for 12 hours, and then anesthetized by intraperitoneal injection of 2% sodium pentobarbital saline. Their body weight was measured, blood was collected from their eyeballs and serum was separated and stored at -80℃. The retroperitoneal fat, greater omentum fat and mesenteric fat were separated and weighed. The following indicators were detected.

[0134] Lipid profile detection in rats: Serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured. The ratios of retroperitoneal fat, greater omentum fat, and mesenteric fat to body weight were calculated.

[0135] The results are as follows: 1) Rat body weight Changes in body weight of rats in each group are as follows: Figure 18 As shown, at the end of the experiment, compared with the control group, the rats in the model group increased their body weight by 14.27%, and the difference was statistically significant. p <0.01 indicates successful model establishment and valid experimental results. Compared with the model group, the EGCG group experienced a 7.45% reduction in body weight ( p <0.05%, TGM group weight loss 3.49% ( p >0.05), the EGCG+TMG mixed group experienced a 5.37% reduction in body weight ( p <0.05%, the supramolecular EGCG-TGM group experienced a 10.61% reduction in body weight (p <0.01), the L-Carn group experienced a weight loss of 8.66% ( p <0.01), the EGCG+L-Carn combination group experienced a 10.95% reduction in body weight ( p <0.01, the supramolecular EGCG-L-Carn group experienced a weight loss of 11.95% ( p <0.01). The body weight of rats in all treatment groups decreased to some extent. Among them, the supramolecular EGCG-TGM group and the supramolecular EGCG-L-Carn group had a more significant ability to reduce the body weight of rats compared with the EGCG group administered alone, the ligand group and the EGCG ligand mixture group. In addition, it is speculated that the supramolecular epigallocatechin gallate has the best ability to reduce the body weight of rats due to its superior bioavailability.

[0136] 2) Blood lipid indicators in rats Blood lipid levels are often evaluated by measuring serum triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and the visceral fat coefficient. Figure 19-23 As shown, compared with the blank group, the model group rats had significantly increased levels of TG, TC, and LDL-C (as shown in the figure). p <0.01), HDL-C significantly decreased ( p <0.01 indicates that the model group rats have lipid metabolism disorders. Compared with the model group, except for the TGM group, the TG, TC, LDL-C, and HDL-C in all groups were significantly improved, indicating that the degree of lipid metabolism disorder in rats was improved to a certain extent. Among them, the supramolecular EGCG-TMG group and the supramolecular EGCG-L-Carn group showed the most significant improvement. The TG, TC, LDL-C, HDL-C, and visceral fat coefficient were not significantly different from those in the blank group. p >0.05).

[0137] (7) Evaluation of anti-photoaging efficacy Using epigallocatechin gallate (EGCG), the supramolecular epigallocatechin gallate-ergothioneine (supramolecular EGCG-EGT) and ergothioneine (EGT) prepared in Example 3 inhibited cell death caused by UVA irradiation, and their anti-photoaging ability was evaluated.

[0138] Press 1×10 4 Human skin fibroblast HFF-1 cells were seeded into 96-well plates at a seeding density of cells / well and incubated at 37°C with 5% CO2. Incubate overnight. When the cell seeding rate in the 96-well plate reaches 40%-60%, add the test substance. Add 100 μL of cell culture medium to each well of the blank group and model group, and add 100 μL of cell culture medium containing the corresponding concentration of the sample to each well of the sample group. The concentration of the test substance is 20 mg / L. The zeroing group has no cell seeding, only 100 μL of cell culture medium. After adding the test substance, place the 96-well plate in an incubator (37℃, 5% CO2) for 24 h. Remove the culture plate, wrap the blank group with aluminum foil, and place it in a cell phototoxicity assay instrument to irradiate with UVA to establish a photodamage model. After irradiation, discard the supernatant, add CCK8 working solution, and incubate at 37℃ in the dark for 2 h. After incubation, read the OD value at 450 nm and calculate the cell viability.

[0139] Cell survival rate results for each group are as follows: Figure 24 As shown, compared with the blank group, the cell survival rate in the model group decreased by 44.33%, and the difference was statistically significant. p <0.01 indicates successful modeling and valid experimental results. Compared to the model group, there was no significant difference in EGCG and EGT ( p >0.05%, presumably due to excessively low concentration. The survival rate of supramolecular EGCG-EGT cells increased by 27.67% compared to the model group ( p The result was <0.01, demonstrating that the supramolecular EGCG-EGT has a more significant ability to inhibit photoaging.

[0140] (8) Evaluation of anti-free radical aging efficacy Using epigallocatechin gallate (EGCG), the supramolecular epigallocatechin gallate-ergothioneine (supramolecular EGCG-EGT) and ergothioneine (EGT) prepared in Example 3 inhibited cellular aging induced by hydrogen peroxide, and their anti-aging capabilities were evaluated.

[0141] Press 2×10 5 Human skin fibroblast HFF-1 cells were seeded into 12-well plates at a seeding density of cells / well and incubated at 37°C with 5% CO2. Incubate overnight. When the cell deposition rate in the 12-well plate reaches 70%-80%, add hydrogen peroxide for induction. Add 1 mL of cell culture medium to each well of the control group, and 1 mL of cell culture medium containing hydrogen peroxide to each well of the model group and sample group. After induction for 4 h, add the corresponding test substance to the sample group at a concentration of 20 mg / L. After adding the test substance, place the 12-well plate in an incubator (37℃, 5% CO2) for 24 h. Remove the culture plate and perform β-galactosidase staining.

[0142] like Figure 25-26As shown, compared with the blank group, the positive cell rate in the model group increased by 60.67%, and the difference was statistically significant (p<0.01), proving that the modeling was successful and the experimental results were effective. Compared with the model group, EGCG, EGT, and supramolecular EGCG-EGT all showed significant differences (p<0.01), with positive cell rates decreasing by 37.11%, 47.67%, and 59.67%, respectively. There was no significant difference between supramolecular EGCG-EGT and the blank group. p >0.05), proving that supramolecular EGCG-EGT has a more significant anti-aging ability.

[0143] The embodiments provided above are not intended to limit the scope of the invention, nor are the described steps intended to limit the order of execution. Any obvious modifications made to the invention by those skilled in the art based on existing common knowledge also fall within the scope of protection defined by the claims.

Claims

1. An EGCG supramolecular material with antioxidant, liver-protective, and fat-reducing effects, characterized in that, It is formed by non-covalent bonding between epigallocatechin gallate and ligands containing trimethylamine and carboxyl groups; the supramolecular has a co-amorphous structure.

2. The supramolecular according to claim 1, characterized in that, The ligands containing trimethylamine and carboxyl groups are betaine, ergothioneine, or L-carnitine.

3. The supramolecular according to claim 1, characterized in that, The molar ratio of epigallocatechin gallate to ligands containing trimethylamine and carboxyl groups is 1:1-2.

4. A method for preparing an EGCG supramolecular compound with antioxidant, liver-protective, and fat-reducing effects as described in any one of claims 1-3, characterized in that, Includes the following steps: Epigallocatechin gallate and a ligand containing trimethylamine and carboxyl groups were dissolved in water and then freeze-dried to obtain the EGCG supramolecular with antioxidant, liver-protective and fat-reducing effects.

5. The preparation method according to claim 4, characterized in that, The ratio of the total mass of epigallocatechin gallate and ligands containing trimethylamine and carboxyl groups to water is 1:2-20 g / mL; the freeze-drying conditions are: pre-freezing at -80℃ for 12-20 h, then transferring to a freeze dryer and drying at -30℃ and 5-10 Pa for 24-30 h, followed by drying at 20℃ and 5-8 Pa for 8 h.

6. A method for preparing an EGCG supramolecular compound with antioxidant, liver-protective, and fat-reducing effects as described in any one of claims 1-3, characterized in that, Includes the following steps: Epigallocatechin gallate and a ligand containing trimethylamine and carboxyl groups were pulverized, mixed, and heated to a molten state. Then, the mixture was immersed in a cooling medium and cooled to a completely solidified state. After cooling to room temperature, it was pulverized and sieved to obtain the EGCG supramolecular with antioxidant, liver-protecting, and fat-reducing effects.

7. The preparation method according to claim 6, characterized in that, Heat to 180-200℃; cool with dry ice or liquid nitrogen; sieve with a mesh size of 80-100.

8. The application of the EGCG supramolecular material with antioxidant, liver-protecting, and fat-reducing effects as described in claim 1, characterized in that, Used in the preparation of pharmaceuticals, food, or daily chemical products.

9. A composition comprising the EGCG supramolecular as described in claim 1, which has antioxidant, liver-protective and fat-reducing effects.

10. The composition according to claim 9, characterized in that, The composition also contains free EGCG, free ligands containing trimethylamine and carboxyl groups, and / or pharmaceutically acceptable excipients.

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