An eye care composition, method of making and use thereof

Abstract

The present application relates to the technical field of daily chemical industry, and in particular to an eye care composition, a preparation method and application thereof.The composition comprises acetyl hexapeptide-8, Bacillus fermentation product, hesperidin methyl chalcone, ergothioneine, ectoine and caffeine.The present application can simultaneously improve periorbital inflammation, microcirculation disorder, lymphatic stasis, glycation damage and barrier fragility, thereby improving problems such as dark circles, eye bags, periorbital fatigue and eyelid laxity.

Description

Technical Field

[0001] This invention relates to the field of daily chemical technology, and in particular to an eye care composition, its preparation method, and its application. Background Technology

[0002] The skin around the eyes is one of the thinnest areas on the face and is most susceptible to physiological changes and environmental influences. Compared to other parts of the face, the stratum corneum around the eyes is thinner, with fewer sebaceous and sweat glands, and weaker support from collagen and elastic fibers in the dermis. However, it also has a rich network of capillaries and lymphatic vessels. Therefore, the area around the eyes is more prone to problems such as dark circles, eye bags, sagging eyelids, fine lines, and a tired appearance.

[0003] Prolonged sleep deprivation, mental stress, exposure to blue light from electronic screens, and environmental stimuli can lead to persistent microcirculatory disturbances, chronic low-grade inflammation, and oxidative stress in the periocular area. Furthermore, increased local vascular permeability, weakened lymphatic drainage, and enhanced glycation promote the deposition of bilirubin, iron ions, and melanin, and accelerate the degradation of the extracellular matrix, resulting in signs of aging such as eye bags, dark circles, and sagging around the eyes.

[0004] Current eye care products typically use simple combinations of caffeine, peptides, hyaluronic acid, plant extracts, or vitamins, primarily targeting single areas such as moisturizing, immediate firming, or reducing fine lines. However, existing technologies still have the following problems:

[0005] (1) Poor stability among active ingredients

[0006] Eye care compositions typically contain peptides, flavonoids, fermentation products, saponins, and antioxidants. However, due to the significant differences in polarity, solubility, and interfacial properties among these different active ingredients, problems such as crystallization, turbidity, sedimentation, color changes, oxidative inactivation, and decreased system viscosity can easily occur.

[0007] For example, hesperidin methyl chalcone is a poorly soluble flavonoid that is easily precipitated in ordinary aqueous systems; caffeine is prone to recrystallization under low-temperature storage conditions; and fermentation products and saponin components may lead to a decrease in the stability of emulsion systems.

[0008] (2) High level of irritation around the eyes

[0009] Traditional plant extracts or saponin-based raw materials often contain highly irritating small molecule impurities and surfactants, which can easily cause eye irritation, redness, tearing, and barrier discomfort. In particular, while horse chestnut-derived active ingredients can improve microcirculation and reduce edema, ordinary alcohol extracts or saponin concentrates have poor long-term tolerance in the delicate area around the eyes.

[0010] (3) The system lacks adaptability to the periocular microenvironment.

[0011] Most existing ophthalmic compositions use common emulsion systems, which make it difficult to ensure the long-term stable coexistence of peptides, fermentation products, flavonoids and antioxidants, and also lack comprehensive consideration of low irritation around the eyes, osmotic pressure compatibility and sustained release performance.

[0012] (4) Difficult to improve multiple periorbital problems simultaneously

[0013] Existing technologies typically address a single problem, such as dark circles, eye bags, or fine lines, making it difficult to simultaneously address multiple factors, including periorbital inflammation, microcirculatory disorders, lymphatic stasis, glycation damage, and a fragile skin barrier.

[0014] Therefore, it is of great significance to develop an eye care composition and its preparation method that can simultaneously improve the stability of active ingredients, reduce periorbital irritation, and improve periorbital inflammation, microcirculation disorders, and fatigue. Summary of the Invention

[0015] This invention provides an eye care composition and its preparation method. This invention also provides a multiphase stable system suitable for the periocular region, which improves the stability and periocular tolerance of peptides, fermentation products, flavonoids, saponins, and antioxidants in the same composition by controlling the distribution state, addition order, and system microenvironment of different active ingredients.

[0016] The present invention adopts the following technical solution: The present invention provides an eye care composition comprising: acetyl hexapeptide-8, Bacillus fermentation product, hesperidin methyl chalcone, ergothioneine, ectoine, and caffeine.

[0017] As a preferred embodiment of the invention, the eye care composition further comprises horse chestnut ferment.

[0018] in:

[0019] Hesperidin methyl chalcone was dispersed in a layered lipid phase formed by hydrogenated lecithin, cholesterol, and ceramide; ectoine and trehalose coexisted in the hydration protection system; acetyl hexapeptide-8, Bacillus fermentation products, and ergothionein were added to the system under low temperature conditions; horse chestnut fermentation products were obtained from horse chestnut raw materials through complex enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, and ultrafiltration fractionation.

[0020] Furthermore, in this invention, hesperidin methyl chalcone is first heated and mixed with hydrogenated lecithin, cholesterol, and oil components to improve its dispersion stability in the system and reduce the risk of crystallization.

[0021] Furthermore, in this invention, caffeine is first dissolved in a trehalose-polyol pre-dissolving system to reduce the risk of recrystallization under low-temperature storage conditions.

[0022] Furthermore, in this invention, ectoine and trehalose jointly regulate the osmotic pressure of the system to improve the stability and periorbital tolerance of fermentation products and peptide active substances.

[0023] Furthermore, in this invention, the horse chestnut raw material is subjected to compound enzymatic hydrolysis and then co-fermented with lactic acid bacteria and brewer's yeast, and the content of irritating small molecule impurities is reduced by ultrafiltration grading, thereby improving the long-term tolerance of use around the eyes.

[0024] Furthermore, in this invention, the pH of the composition is controlled at 6.2 to 6.8, and the osmotic pressure is controlled at 280 to 320 mOsm / kg.

[0025] Furthermore, in this invention, the composition is used to improve eye bags, dark circles, eye fatigue, eyelid sagging, and eyelid aging induced by lack of sleep.

[0026] To solve the above-mentioned technical problems, the present invention provides a method for preparing an eye care composition, comprising the following steps:

[0027] (1) Add trehalose, ectoine, glycerol, butanediol and metal ion complexing agent to the aqueous phase and mix and dissolve them, then add to the thickening system for hydration;

[0028] (2) Hesperidin methyl chalcone was heated together with hydrogenated lecithin, cholesterol, ceramide and oil components to disperse hesperidin methyl chalcone in the layered lipid structure;

[0029] (3) Add the flavonoid lipid-encapsulated phase obtained in step (2) to step (1) to form a liquid crystal layered emulsion system;

[0030] (4) Caffeine is pre-dissolved in a trehalose-polyol pre-dissolved system and then added to the system obtained in step (3);

[0031] (5) When the system temperature drops to below 35-40℃, add acetyl hexapeptide-8, Bacillus fermentation product, horse chestnut fermentation product and ergothioneine;

[0032] (6) Adjust the pH of the system to 6.2-6.8 and control the osmotic pressure to 280-320 mOsm / kg to obtain the eye care composition;

[0033] The horse chestnut ferment is prepared from horse chestnut raw material through compound enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, and ultrafiltration fractionation.

[0034] Preferably, the method for preparing the horse chestnut ferment includes the following steps:

[0035] (1) Crush the horse chestnut raw material and add it to water;

[0036] (2) Add cellulase and pectinase for compound enzymatic hydrolysis;

[0037] (3) After enzyme inactivation, inoculate with Lactobacillus plantarum and Saccharomyces cerevisiae for synergistic fermentation;

[0038] (4) Add trehalose and ectoine during fermentation;

[0039] (5) After fermentation, perform ultrafiltration fractionation;

[0040] (6) Collect the permeate and mix it with part of the retentate to obtain horse chestnut ferment.

[0041] The present invention also provides an eye care composition prepared by the aforementioned preparation method.

[0042] An eye care product comprising the aforementioned eye care composition.

[0043] Preferably, the eye care product includes:

[0044] Acetyl hexapeptide-8; Bacillus fermentation product; Aesculus hippocastanum fermentation product; Hesperidin methyl chalcone; Ergothioneine; Ectocin; Caffeine.

[0045] Preferably, the eye care product comprises the following components by weight percentage:

[0046] Trehalose: 0.5–5.0%; Ectoin: 0.05–1.0%; Glycerin: 1.0–10.0%; Butylene glycol: 1.0–10.0%; Caffeine: 0.05–1.0%; Acetyl hexapeptide-8: 0.1–10.0%; Bacillus fermentation product: 0.1–10.0%; Horse chestnut fermentation product: 0.1–10.0%; Hesperidin methyl chalcone: 0.001–1.0%; Ergothioneine: 0.001–1.0%; Dipotassium glycyrrhizate: 0.01–1.0%; Acetylated sodium hyaluronate: 0.001–1.0%; Sodium hyaluronate: 0.001–1.0%; Hydrogenated lecithin: 0.1– 5.0%; Ceramide NP: 0.001–1.0%; Cholesterol: 0.001–1.0%; Phytosterol: 0.001–2.0%; Squalane: 0.1–10.0%; C13–15 Alkanes: 0.1–10.0%; Caprylic / Capric Triglycerides: 0.1–10.0%; Polyglycerol Fatty Acid Emulsifier: 0.05–5.0%; Hydroxyethyl Cellulose: 0.05–2.0%; Acrylic (Ester) / C10–30 Alkyl Acrylate Crosspolymer: 0.01–2.0%; Metal Ion Complexing Agent: 0.001–0.5%; Preservative System: 0.1–3.0%; Deionized Water: Balance.

[0047] Preferably, the pH of the composition is 6.2 to 6.8.

[0048] Preferably, the composition is used to improve one or more of the following: eye bags, dark circles, eyelid sagging, eye fatigue, and sleep deprivation-induced periorbital aging.

[0049] Preferably, the eye care composition is used in the preparation of eye care products for regulating periorbital inflammation, microcirculation, lymphatic integrity and barrier stability.

[0050] Preferably, the regulation includes one or more of the following: IL-33 / ST2 inflammatory pathway regulation, VEGF / VE-Cadherin permeability regulation, Emilin-1 / Integrin α9β1 lymphoid integrity regulation, AGEs / ALEs glycation regulation, and desmosome and tight junction enhancement.

[0051] Preferably, the eye care product is one or more of the following: eye cream, eye serum, eye mask, liquid crystal emulsion, gel, and microcapsule sustained-release formulation.

[0052] By adopting the above technical solution, the present invention has the following beneficial effects:

[0053] (1) Improve the stable coexistence of multiple active ingredients

[0054] This invention improves the stability of different types of active ingredients in the same ocular system by controlling the distribution and addition order of flavonoids, peptides, fermentation products and antioxidant active ingredients, and reduces the risk of crystallization, turbidity, oxidative inactivation and system instability.

[0055] (2) Improve the dispersion stability of flavonoid active substances

[0056] In this invention, hesperidin methyl chalcone, hydrogenated lecithin, cholesterol, and ceramide together form a layered lipid phase, thereby improving its dispersion stability and reducing the risk of crystallization and periorbital irritation.

[0057] (3) Reduce the irritant properties of active ingredients derived from horse chestnut

[0058] This invention employs compound enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, and ultrafiltration fractionation to process horse chestnut raw materials, thereby reducing the content of irritating small molecules and surface-active impurities in traditional horse chestnut extracts and improving long-term tolerance around the eyes.

[0059] (4) Improve the stability of peptides and fermentation products

[0060] In this invention, acetyl hexapeptide-8, Bacillus fermentation products, and ergothioneine are added to the system under low temperature conditions, thereby reducing the risk of thermal degradation and oxidative inactivation.

[0061] (5) Reduce the risk of caffeine crystallization

[0062] This invention uses a trehalose-polyol pre-dissolving system to improve the solubility stability of caffeine, thereby reducing the risk of caffeine recrystallization under low-temperature storage conditions.

[0063] (6) Improve long-term tolerance around the eyes

[0064] This invention utilizes ectoine, trehalose, and an osmotic pressure control system to make the composition more suitable for long-term use in the delicate periocular area, thereby reducing irritation and improving comfort.

[0065] (7) Improves multidimensional periorbital problems

[0066] This invention can simultaneously improve periorbital inflammation, microcirculation disorders, lymphatic retention, glycation damage, and barrier fragility, thereby improving problems such as dark circles, eye bags, periorbital fatigue, and eyelid sagging. Attached Figure Description

[0067] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0068] Preparation of Figures 1-3 The images show the changes in crow's feet before (0 days), after (14 days), and after (28 days) the use of the product in Example 1 by subjects A to C. Detailed Implementation

[0069] The present invention will be further explained below with reference to specific embodiments.

[0070] This invention relates to a cosmetic composition suitable for use in the periorbital area and its preparation method. The composition is not a simple mixture of multiple periorbital active ingredients, but rather addresses common problems in periorbital formulations such as poor solubility of flavonoids, low-temperature crystallization of caffeine, insufficient thermal stability of peptides and fermentation products, high irritation of horse chestnut-derived active ingredients, and incompatible osmotic pressure in periorbital systems. It provides a stable eye care system with multiphase distribution characteristics.

[0071] In this invention, the composition is preferably in the form of an eye cream, eye serum, eye gel, liquid crystal emulsion, or eye mask serum. The composition is suitable for improving dark circles, eye bags, eye fatigue, eyelid sagging, and fine lines around the eyes caused by insufficient sleep, stress, microcirculation disorders, weakened skin barrier, glycation, and oxidative stress.

[0072] I. Definition of Terms

[0073] For ease of understanding of this invention, unless otherwise stated, the following terms have the following meanings.

[0074] The "periocular area" refers to the skin around the eyelids, under the eyes, the outer corners of the eyes, the periorbital area, and adjacent skin. This area has a thinner stratum corneum, less sebum secretion, and a rich network of capillaries and lymphatic vessels, making it more sensitive to irritating ingredients, changes in osmotic pressure, pH changes, and instability of emulsion systems.

[0075] "Multiphase stable system" refers to a composition in which different active ingredients are not all in the same homogeneous aqueous phase, but are distributed in the hydration protection phase, lipid-encapsulated phase, fermentation active phase and low-temperature antioxidant phase according to the polarity, solubility, thermal stability and irritation of the active ingredients, thereby reducing the mutual interference between different active ingredients.

[0076] The "osmolar hydration protective phase" refers to a hydration environment formed by trehalose, ectoine, polyols, and water. This phase can improve the system's tolerance to electrolytes, fermentation products, and peptide actives through hydrogen bonding and water molecule organization, while reducing periorbital irritation.

[0077] "Laminated lipid phase" refers to lipid microdomains with a tendency to be arranged in layers, formed by hydrogenated lecithin, cholesterol, ceramide, phytosterols, and lipid components. This structure can improve the dispersion stability of poorly soluble flavonoids in the system and reduce the risk of crystallization or irritation caused by their direct exposure to the aqueous phase.

[0078] "Flavone lipid-encapsulated phase" refers to the structural state in which flavonoid active substances such as hesperidin methyl chalcone are dispersed or embedded in a layered lipid phase.

[0079] "Aesculus horseii ferment" refers to the fermentation product obtained from aesculus horseii raw material through enzymatic hydrolysis, co-fermentation with lactic acid bacteria and yeast, ultrafiltration fractionation, and hydration stabilization treatment. This ferment differs from ordinary aesculus horseii alcohol extract or saponin concentrate; it has a lower content of irritating small molecule impurities, making it more suitable for use around the eyes.

[0080] The "low-temperature antioxidant active phase" refers to the active phase added after the preparation system has been cooled to a lower temperature. It contains heat-sensitive or easily oxidized active substances such as acetyl hexapeptide-8, Bacillus fermentation products, and ergothioneine. The purpose of this phase is to reduce the damage to the structure of the active substances caused by high temperature and high shear.

[0081] "Tear-like microenvironment" refers to the pH and osmotic pressure of the composition being controlled within a range that is close to the tolerance of the eye, preferably with a pH of 6.2 to 6.8 and an osmotic pressure of 280 to 320 mOsm / kg, in order to reduce eye irritation, tearing and discomfort.

[0082] Current eye care products typically incorporate caffeine, peptides, plant extracts, hyaluronic acid, and antioxidants directly into conventional emulsification systems. While this approach allows for ingredient blending, it has significant limitations in eye formulations.

[0083] First, the solubility of different active ingredients varies considerably. Caffeine is a small, water-soluble molecule, but its solubility decreases under low-temperature storage conditions, easily forming small crystals. Hesperidin methyl chalcone is a flavonoid derivative with poor solubility in aqueous phase; if added directly to ordinary hydrogels or creams, it easily causes crystallization, turbidity, or precipitation. Acetyl hexapeptide-8 and Bacillus fermentation products are mostly located in the aqueous phase, but they are sensitive to high temperatures, metal ions, pH fluctuations, and microbial metabolic impurities, easily leading to a decrease in activity retention.

[0084] Secondly, while active ingredients derived from horse chestnut have the potential to improve periorbital edema and microcirculation, common horse chestnut extracts may contain a relatively high amount of saponins, tannins, and low-molecular-weight irritants. Saponins have certain interfacial activity; when directly added to periorbital emulsion systems, they may alter the emulsion membrane structure, leading to decreased system viscosity, increased foaming, or enhanced periorbital irritation.

[0085] Secondly, the skin around the eyes is more sensitive to changes in osmotic pressure and pH. Regular facial creams may not be suitable for the eye area, especially when they contain fermentation products, salts, saponins, or high concentrations of polyols. The conductivity, osmotic pressure, and free water state of the system may change, thus causing discomfort around the eyes.

[0086] The eye care composition of the present invention preferably comprises the following functional components:

[0087] 1. Trehalose, ectoine, glycerin, butylene glycol, and panthenol

[0088] Trehalose has a strong hydration effect and can form a stable hydration environment with water molecules and hydrophilic active substances through multi-point hydrogen bonds. Ectoin is a cyclic amino acid derivative that has the function of stabilizing the hydration layer around proteins and cell membranes. In this invention, ectoin is used not only as a soothing active substance but also as a hydration stabilizer to regulate the system's microenvironment.

[0089] Glycerin and butylene glycol, as polyols, can improve the system's moisturizing properties and assist in the pre-dissolution of caffeine. Panthenol can improve the comfort of the periorbital barrier and, together with trehalose and ectoine, reduce the irritation of the periorbital area caused by fermentation products and saponins.

[0090] In the preferred formulation, the amount of trehalose is 0.5-5.0%, the amount of ectoine is 0.05-1.0%, the amount of glycerol is 1.0-10.0%, the amount of butylene glycol is 1.0-10.0%, and the amount of panthenol is 0.1-3.0%.

[0091] 2. Peptides and fermentation active components

[0092] The peptides and fermentation-active components include acetyl hexapeptide-8 and Bacillus fermentation products.

[0093] Acetyl hexapeptide-8 is a hydrophilic polypeptide suitable for addition in a low-temperature aqueous phase. Prolonged exposure to high temperatures or strong shearing processes may lead to decreased structural stability. Therefore, this invention uses it as a post-addition active ingredient. Bacillus fermentation products contain fermentation metabolites, small peptides, polysaccharides, or amino acids, which can help improve periorbital fatigue and barrier function. However, these fermentation products typically contain electrolytes or low-molecular-weight metabolites, and direct addition to a conventional carbomer system may cause instability in the thickening system. Therefore, this invention first creates a trehalose-ectoine hydration protective environment before adding the fermentation products at low temperatures to reduce system mutation.

[0094] 3. Flavonoid microcirculation active components

[0095] The preferred flavonoid microcirculation active component is hesperidin methyl chalcone. This component has the potential to improve microcirculation around the eyes and reduce pigmented and vascular dark circles, but its water solubility is poor, and it is prone to crystallization in conventional eye creams. In this invention, hesperidin methyl chalcone is pre-dispersed in a layered lipid phase formed by hydrogenated lecithin, cholesterol, ceramide NP, phytosterols, and oil components. This treatment reduces direct contact between hesperidin methyl chalcone and the aqueous phase, lowers the risk of crystallization, and allows it to exist in a more stable lipid microdomain form.

[0096] 4. Caffeine stabilizing components

[0097] Caffeine can be used to improve puffiness around the eyes and a tired appearance, but it poses a risk of crystallization at low temperatures. This invention preferably involves dissolving caffeine in a trehalose-polyol pre-dissolving system before adding it to the main system. The polyol enhances the solubility and retention of caffeine in the aqueous phase, while trehalose reduces the tendency of caffeine molecules to rearrange and form crystals at low temperatures through its hydration structure. The preferred amount of caffeine is 0.05–1.0%, more preferably 0.10–0.50%.

[0098] 5. Antioxidant stabilizing components

[0099] The antioxidant stabilizing component preferably includes ergothioneine. Ergothioneine is sensitive to metal ions, oxidative environments, and high temperatures. To improve its activity retention, this invention adds ergothioneine together with acetyl hexapeptide-8 and Bacillus fermentation products at a low temperature. Furthermore, disodium EDTA, sodium phytate, or a combination thereof can be added to the system to chelate trace metal ions such as Fe and Cu, reducing metal ion-induced oxidative discoloration and activity loss.

[0100] 6. Horse chestnut ferment

[0101] In this invention, the active ingredient derived from horse chestnut is preferably not a common alcohol extract, but a horse chestnut ferment.

[0102] The problem with common horse chestnut extract is that its saponin components have strong interfacial activity and may contain tannins, free small-molecule phenolic acids, and other irritating impurities. For the eye area, these impurities may increase the risk of stinging and tearing.

[0103] This invention employs a combination of enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, and ultrafiltration fractionation to process horse chestnut raw materials. Enzymatic hydrolysis gently releases saponins, polysaccharides, and polyphenol precursors from horse chestnut; lactic acid bacteria fermentation reduces some irritating impurities and generates milder small-molecule metabolites; yeast fermentation provides polysaccharides, amino acids, and antioxidant metabolites; and ultrafiltration fractionation further reduces the content of free small-molecule irritants while retaining some of the polysaccharide-saponin-peptide complex components.

[0104] In the preferred embodiment, trehalose II and ectoine II (which are not the same as the trehalose and ectoine in the eye care composition) are added during the fermentation process, so that the horse chestnut-derived active ingredients are in a hydrated and protected environment during the fermentation stage, thereby reducing irritation from the raw material end and improving compatibility with the periorbital formulation system.

[0105] Horse chestnut ferment is prepared as follows:

[0106] The seeds of *Aesculus hippocastanum* are shelled, dried, and then pulverized to 40–100 mesh, preferably 60–80 mesh. Deionized water is added at a material-to-liquid ratio of 1:8–1:20, preferably 1:12. The pH is adjusted to 5.2–6.0, preferably 5.6, using a citric acid-sodium citrate buffer system. Cellulase and pectinase are added to the system for combined enzymatic hydrolysis. The preferred dosage of cellulase is 0.05–0.50%, and the preferred dosage of pectinase is 0.05–0.30%. The hydrolysis temperature is 40–50°C, and the hydrolysis time is 1–3 hours. Through enzymatic hydrolysis, the cell wall structure of *Aesculus hippocastanum* is partially disrupted, which facilitates the gentle release of active ingredients while avoiding the instantaneous accumulation of saponins and increased irritation caused by strong alcohol extraction.

[0107] After enzymatic hydrolysis, the system is heated to 85–95°C and held for 5–15 minutes to inactivate the enzymes. It is then cooled to 28–35°C and inoculated with *Lactobacillus plantarum* and *Saccharomyces cerevisiae*. The preferred inoculum size for *Lactobacillus plantarum* is 1.0 × 10⁻⁶. 6 ~1.0×10 8 The optimal inoculum size for *Saccharomyces cerevisiae* is 1.0 × 10⁻⁶ CFU / mL. 5 ~1.0×10 7 CFU / mL.

[0108] Trehalose II and ectoine II are added during fermentation. The preferred dosage of trehalose is 0.5–5.0%, and the preferred dosage of ectoine is 0.05–1.0%. The fermentation temperature is 28–35℃, and the fermentation time is 24–48 hours.

[0109] After fermentation, the pH of the system is adjusted to 5.8–6.5, and insoluble matter is removed by centrifugation. The supernatant is then subjected to ultrafiltration fractionation. The preferred molecular weight cutoff of the ultrafiltration membrane is 1000–5000 Da, more preferably 3000 Da. Ultrafiltration can remove some free low-molecular-weight irritating substances while retaining polysaccharide, peptide, and saponin complex components with good hydration stability.

[0110] In a preferred embodiment, the ultrafiltration permeate and a portion of the retained solution are mixed at a mass ratio of 5:5 to 9:1, preferably 7:3. This treatment avoids the complete removal of the high molecular weight protective components while reducing the proportion of irritating small molecules.

[0111] The preferred preparation process of the present invention includes the following steps.

[0112] 1. Preparation of hydrated protective phase

[0113] Add deionized water to the main pot, and add glycerin, butylene glycol, trehalose, ectoine, panthenol and metal ion complexing agent at 25-45°C, and stir until dissolved.

[0114] Subsequently, hydroxyethyl cellulose and acrylate / C10-30 alkanol acrylate crosspolymers are added, stirred, and allowed to stand for hydration. The purpose of this step is to first form a hydrated gel network with certain electrolyte tolerance. If fermentation products, horse chestnut ferment, or electrolyte components are added before the gel network is formed, the hydration of the thickener may be interfered with, leading to unstable viscosity or subsequent water separation.

[0115] 2. Preparation of flavonoid lipid phase

[0116] In a separate oil phase pot, add hydrogenated lecithin, cholesterol, ceramide NP, phytosterols, squalane, C13-15 alkanes, caprylic / capric triglycerides, and hesperidin methyl chalcone. Heat the system to 68–78°C, preferably 70–75°C, and maintain the temperature while stirring until the lipid components are fully melted and dispersed.

[0117] The technical purpose of this step is to ensure that hesperidin methyl chalcone is fully dispersed in a high-temperature lipid environment and enters the lamellar lipid microdomains during subsequent emulsification. If hesperidin methyl chalcone is added directly to a cooled aqueous phase or a conventional emulsion system, its molecules are difficult to disperse uniformly and are prone to crystallization after low-temperature storage.

[0118] 3. Emulsification

[0119] The flavonoid lipid phase was slowly added to the hydrated protective phase, and homogenized at 3000–6000 rpm for 2–8 minutes to obtain a liquid crystal layered emulsion system. The system was then cooled by low-speed stirring.

[0120] In this step, hydrogenated lecithin, cholesterol, and ceramide NP synergistically form a layered arrangement, mimicking the lipid structure of the skin barrier and providing stable dispersion sites for insoluble flavonoids. This structure also reduces the irritation caused by direct contact of the active ingredient with the skin around the eyes.

[0121] 4. Add caffeine pre-solution

[0122] Caffeine is pre-dissolved in a partial aqueous solution of butylene glycol, glycerol, and trehalose, and stirred at 45–55°C until clear. Once the main system has cooled to below 45°C, the pre-dissolved caffeine solution is added to the main system.

[0123] This step reduces the risk of caffeine forming crystals under cold storage conditions. If caffeine is added directly to a low-temperature emulsion system in powder form, undissolved particles may remain due to excessively high local concentrations, which can then induce further recrystallization.

[0124] 5. Addition of low-temperature active ingredients

[0125] When the system is cooled to below 35-40℃, preferably below 35℃, acetyl hexapeptide-8, Bacillus fermentation product, horse chestnut ferment, ergothioneine, sodium hyaluronate components and dipotassium glycyrrhizate are added in sequence.

[0126] The low-temperature addition sequence is an important process feature of this invention. Acetyl hexapeptide-8 can reduce thermal degradation; Bacillus fermentation products can reduce color changes or activity loss caused by high temperatures; ergothioneine can avoid prolonged heating and metal ion-induced oxidation; the subsequent addition of horse chestnut fermentation products can reduce the interference of its saponin components on the emulsion film formation process.

[0127] 6. pH and osmotic pressure adjustment

[0128] Finally, the pH of the system was adjusted to 6.2–6.8, preferably 6.4 ± 0.2, using arginine, aminomethylpropanol, citric acid, or a combination thereof. The osmotic pressure was measured and controlled at 280–320 mOsm / kg.

[0129] The reason for pH control is that the periorbital area is sensitive to acidic or alkaline systems. Too low a pH may increase irritation and affect the stability of some peptides and fermentation products; too high a pH may accelerate color changes in certain flavonoids and antioxidants. Osmotic pressure control reduces the osmotic pressure difference between the composition and the periorbital microenvironment, thereby improving comfort.

[0130] Trehalose and ectoine stabilize the aqueous active substances through hydration. They alter the free water state, forming a more stable hydrated layer and reducing the damage to the gel network caused by electrolytes, fermentation metabolites, and saponins. This hydrated environment also reduces conformational disturbances of peptides and fermentation products during storage. Hesperidin methyl chalcone exhibits strong hydrophobicity and a tendency to crystallize, easily precipitating when alone in the aqueous phase. This invention utilizes hydrogenated lecithin, cholesterol, ceramide, and lipids to form lipid microdomains, distributing hesperidin methyl chalcone within the lipid structure, thereby reducing its supersaturation in the aqueous phase. A pre-dissolved caffeine system reduces local supersaturation. Low-temperature caffeine crystallization is usually related to excessively high local concentrations and changes in the solvent environment. This invention uses polyols and trehalose for pre-dissolution, ensuring caffeine is uniformly dissolved before being added to the main system, thus reducing crystal nucleus formation. Combined enzymatic hydrolysis and fermentation alter the release state of horse chestnut-derived active substances, and ultrafiltration fractionation further reduces free, irritating small molecules. Trehalose and ectoine participate in hydration protection during the fermentation stage, making horse chestnut fermentation products more compatible with periorbital formulations. Peptides, fermentation products, and ergothioneine are not suitable for prolonged high-temperature processing. By adding them at a low temperature after emulsification, the risks of thermal degradation, oxidative inactivation, and color changes can be significantly reduced.

[0131] The eye care composition of the present invention is suitable for eye area care, especially for problems such as eye bags, dark circles, eye fatigue, eyelid sagging and fine lines caused by insufficient sleep, long-term staying up late, mental stress, microcirculation disorders, oxidative stress, enhanced glycation reaction and barrier fragility.

[0132] The composition of the present invention not only improves a single periocular problem, but also improves the long-term fatigue and aging state of the periocular region from multiple directions by comprehensively regulating local inflammation, microcirculation, lymphatic function, glycation status and barrier integrity.

[0133] In a preferred embodiment, the composition of the present invention can be used to improve sleep deprivation-induced eye fatigue; improve vascular and pigmented dark circles around the eyes; improve eye puffiness and eye bags; improve eyelid laxity; improve fine lines around the eyes; enhance the tolerance of the eye barrier; and improve dark circles and local microcirculation disorders caused by long-term sleep deprivation.

[0134] This invention posits that periorbital fatigue is not solely caused by insufficient hydration or skin laxity, but is related to multiple biological processes, including inflammation, vascular permeability, lymphatic retention, glycation, and decreased barrier function. Therefore, this invention improves the overall condition of the periorbital area by regulating multiple related pathways.

[0135] I. Regulation of the IL-33 / ST2 Inflammatory Pathway

[0136] In this invention, the "IL-33 / ST2 inflammatory pathway" refers to an inflammatory signaling pathway involving interleukin-33 (IL-33) and its receptor ST2. IL-33 belongs to the class of alarmin cytokines. Cells can release IL-33 when the skin is subjected to stress, sleep deprivation, oxidative stress, ultraviolet radiation stimulation, or inflammatory damage. After IL-33 binds to its receptor ST2, it can further induce the release of inflammatory mediators and promote increased vascular permeability, pigment production, and local inflammatory responses.

[0137] In the periorbital area, long-term sleep deprivation and chronic fatigue can easily lead to a sustained increase in IL-33, resulting in: enhanced periorbital inflammatory response; vasodilation; increased melanin production; abnormal microcirculation; local edema; and decreased barrier integrity.

[0138] In this invention, acetyl hexapeptide-8, Bacillus fermentation products, ergothioneine, and horse chestnut fermentation products can reduce local inflammatory stimulation and decrease IL-33-related inflammatory signals, thereby reducing chronic low-grade inflammation around the eyes.

[0139] Furthermore, the hydration environment formed by trehalose and ectoine can reduce the disturbance of the periocular barrier by active substances and external stimuli, thereby indirectly reducing inflammation induction. Therefore, this invention can improve sleep-deprivation-induced periocular fatigue, dark circles, and chronic irritation by regulating the IL-33 / ST2 inflammatory pathway.

[0140] II. Regulation of VEGF / VE-Cadherin permeability

[0141] In this invention, "VEGF / VE-Cadherin permeability regulation" refers to the regulation of vascular permeability related to vascular endothelial growth factor (VEGF) and vascular endothelial cadherin (VE-Cadherin).

[0142] VEGF is an important signaling molecule that promotes angiogenesis and increased vascular permeability. VEGF expression may increase when local inflammation, hypoxia, or sleep deprivation is present, leading to: capillary dilation, increased vascular permeability, increased tissue fluid exudation, local edema, eye bag formation, and deposition of heme degradation products.

[0143] VE-Cadherin is an important adhesion protein present between vascular endothelial cells, participating in maintaining vascular endothelial integrity and the stability of intercellular connections. When VE-Cadherin is stable, the intercellular connections between vascular endothelial cells are tighter, and tissue fluid leakage is reduced.

[0144] In this invention, hesperidin methyl chalcone, horse chestnut ferment, and caffeine can improve local microcirculation and reduce local vascular permeability. Horse chestnut-derived active ingredients can reduce local tissue fluid accumulation, while hesperidin methyl chalcone helps improve microvascular stability.

[0145] Furthermore, by reducing inflammatory stimulation and oxidative stress, the present invention can indirectly reduce the abnormal increase of VEGF, thereby reducing local edema and the formation of eye bags.

[0146] Therefore, this invention can improve periorbital puffiness, eye bags, and vascular dark circles by regulating VEGF / VE-Cadherin-related permeability.

[0147] III. Regulation of Lymphatic Integrity by Emilin-1 / Integrin α9β1

[0148] In this invention, "Emilin-1 / Integrin α9β1 lymphatic integrity regulation" refers to the regulation of lymphatic-related extracellular matrix proteins and the lymphatic endothelial adhesion system.

[0149] Emilin-1 (Elastin Microfibril Interface Located Protein-1) is an extracellular matrix glycoprotein associated with the stability of elastic fibers and lymphatic structures. It participates in maintaining the integrity of lymphatic vessels and the return of tissue fluid.

[0150] Integrin α9β1 belongs to the integrin family and is involved in lymphatic endothelial cell adhesion, lymphatic vessel stability, and lymphatic fluid transport.

[0151] The area around the eyes has a rich but relatively fragile lymphatic network. When there is insufficient sleep, inflammation, or microcirculatory disturbances, local lymphatic drainage weakens, and tissue fluid and inflammatory metabolites easily accumulate, resulting in eye bags, puffiness, local dark circles, and a chronically fatigued appearance.

[0152] In this invention, horse chestnut ferment, Bacillus ferment, and hesperidin methyl chalcone can improve local microcirculation and tissue fluid metabolism. Furthermore, the low-irritation hydration environment formed by trehalose and ectoine helps maintain the stability of the periocular barrier and reduces the interference of long-term local inflammation on lymphatic structures.

[0153] Therefore, the present invention can improve the state of tissue fluid retention in the periorbital area and enhance the local metabolic capacity under long-term fatigue in the periorbital area.

[0154] IV. Glycation Regulation of AGEs / ALEs

[0155] In this invention, "AGEs / ALEs glycation regulation" refers to the regulation of the formation process of advanced glycation end products (AGEs) and advanced lipid peroxidation end products (ALEs).

[0156] AGEs are end products formed from non-enzymatic reactions of carbohydrates with proteins, lipids, or nucleic acids. ALEs, on the other hand, are related to lipid oxidation.

[0157] Chronic sleep deprivation, stress, inflammation, and oxidative stress can accelerate the formation of AGEs and ALEs, leading to stiff collagen fibers, decreased elasticity, dull skin, ECM degradation, sagging around the eyes, and increased fine lines. The skin around the eyes is thinner and more susceptible to glycation damage. In this invention, ergothioneine, Bacillus fermentation products, acetyl hexapeptide-8, and trehalose can reduce oxidative stress and free radical damage, thereby reducing the formation of AGEs and ALEs.

[0158] Ergothioneine, as a sulfur-containing antioxidant, can reduce oxidation-induced lipid peroxidation. Trehalose can stabilize the hydration state around proteins, thereby reducing the risk of protein glycation. Small molecule metabolites and antioxidant components in Bacillus fermentation products can also help reduce glycation damage.

[0159] Therefore, the present invention can improve the appearance of dark circles, sagging and fatigue around the eyes caused by glycation and oxidative stress.

[0160] In this invention, "desmosome and tight junction strengthening" refers to improving the stability of cell connections and barrier integrity between keratinocytes. Desmosomes are intercellular junction structures that participate in maintaining the mechanical stability between keratinocytes. Desmosome-related proteins include JUP (junction plakoglobin), DSP (desmoplakin), PKP (plakophilin), and DSG (desmoglein). Tight junctions are involved in regulating intercellular barrier permeability, and related proteins include Claudin, Occludin, and ZO-1.

[0161] The area around the eyes has a thinner stratum corneum and a weaker barrier stability, making it more susceptible to irritating ingredients, inflammation, and environmental stress. When desmosomes and tight junctions are damaged, the area around the eyes is more prone to redness, irritation, dryness, moisture loss, and chronic sensitivity.

[0162] In this invention, ectoine, trehalose, ceramide, cholesterol, and Bacillus fermentation products can improve the stability of the periocular barrier. Ceramide and cholesterol can improve the integrity of the lipid barrier; trehalose and ectoine can stabilize the hydration environment of the stratum corneum; and fermentation products can improve the stability of the local microenvironment.

[0163] Furthermore, the layered lipid phase exhibits good biocompatibility with the lipid structure of the periocular skin, thereby improving long-term tolerance to use around the eyes. Therefore, this invention can enhance the stability of the periocular barrier, reduce irritation, and alleviate periocular fragility under prolonged fatigue.

[0164] In a preferred embodiment, the eye care product comprises, by weight percentage:

[0165] Trehalose 0.5–5.0%, Ectoin 0.05–1.0%, Glycerin 1.0–10.0%, Butylene Glycol 1.0–10.0%, Caffeine 0.05–1.0%, Acetyl Hexapeptide-8 Solution 0.1–10.0%, Bacillus Fermentation Product 0.1–10.0%, Horse Chestnut Ferment 0.1–10.0%, Hesperidin Methyl Chalcone 0.001–1.0%, Ergothioneine 0.001–1.0%, Dipotassium Glycyrrhizate 0.01–1.0%, Acetylated Sodium Hyaluronate 0.001–1.0%, Sodium Hyaluronate 0.001–1.0%, Hydrogenated Lecithin 0.1–5%. 0%, Ceramide NP 0.001-1.0%, Cholesterol 0.001-1.0%, Phytosterol 0.001-2.0%, Squalane 0.1-10.0%, C13-15 Alkanes 0.1-10.0%, Caprylic / Capric Triglycerides 0.1-10.0%, Polyglycerol Fatty Acid Emulsifier 0.05-5.0%, Hydroxyethyl Cellulose 0.05-2.0%, Acrylic (Ester) / C10-30 Alkyl Acrylate Crosspolymer 0.01-2.0%, Metal Ion Complexing Agent 0.001-0.5%, Preservative System 0.1-3.0%, Deionized Water to 100%.

[0166] Example 1

[0167] This embodiment provides an eye care product, which, by weight percentage, is composed of the following components:

[0168] Deionized water, balance; Glycerin 4.50%; Butylene glycol 3.50%; Trehalose I 2.50%; Ectoin I 0.30%; Panthenol 1.00%; Caffeine 0.30%; Acetyl hexapeptide-8 solution 5.00%; Bacillus fermentation product 3.00%; Horse chestnut fermentation protective complex 4.00%; Hesperidin methyl chalcone 0.05%; Ergothioneine 0.10%; Dipotassium glycyrrhizate 0.20%; Acetylated sodium hyaluronate 0.05%; Sodium hyaluronate 0.03%; Hydrogenated lecithin 1.50%; Ceramide NP 0.15%; Cholesterol 0.10%; Phytosterol 0.20%; Squalane 3.00%; C13-15 Alkanes 4.00%; Caprylic / Capric Triglyceride 3.00%; Polyglycerol-10 Stearate 0.60%; Hydroxyethyl Cellulose 0.25%; Acrylic (Ester) / C10-30 Alkyl Acrylate Crosspolymer 0.18%; Disodium EDTA 0.03%; Sodium Phytate 0.02%; 1,2-Hexanediol 0.50%; Ethylhexylglycerin 0.30%; Arginine as needed.

[0169] Trehalose I and ectoine I are functional components added later in the eye care composition. Trehalose II and ectoine II contained in the horse chestnut fermentation protective complex are derived from the horse chestnut fermentation process and are not included in the calculation with trehalose I and ectoine I.

[0170] Preparation of Horse Chestnut Fermentation Protective Complex

[0171] Take horse chestnut seeds, remove the shells, dry at 45℃ until the moisture content is below 8%, and pulverize to 60 mesh to obtain horse chestnut powder. Weigh 100 g of horse chestnut powder, add 1200 g of deionized water, and adjust the pH to 5.60 using a citric acid-sodium citrate buffer system.

[0172] Add 2.0 g of cellulase and 1.0 g of pectinase to the above system, and hydrolyze at 45°C for 1.5 hours with a stirring speed of 300 rpm. After hydrolysis, heat the system to 90°C and incubate for 10 minutes to inactivate the enzyme, then cool to 32°C.

[0173] Lactobacillus plantarum and Saccharomyces cerevisiae were inoculated into the cooled enzymatic hydrolysate, with an inoculum size of 1.0 × 10⁻⁶. 7 CFU / mL, Saccharomyces cerevisiae inoculum size 1.0 × 10⁻⁶ 6 CFU / mL. Trehalose II 20.0 g and ectoine II 3.0 g were added simultaneously, and fermentation was carried out at 32℃ for 36 hours. During fermentation, the pH was controlled at 5.4-5.8, and the stirring speed was 150 rpm.

[0174] After fermentation, the system was adjusted to pH 6.0 and centrifuged at 4000 rpm for 15 minutes. The supernatant was collected. The obtained supernatant was subjected to ultrafiltration fractionation using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. The permeate and a portion of the retentate were collected and mixed at a mass ratio of 7:3. Ergothioneine (0.05%) and dipotassium glycyrrhizate (0.10%) were added to the mixture, and the mixture was filtered through a 0.22 μm filter to obtain the horse chestnut fermentation protective complex.

[0175] Eye care products Appearance and centrifuge stability testing

[0176] Add deionized water to the mixing bowl and heat to 35°C. Add glycerol, butylene glycol, trehalose I, ectoine I, panthenol, disodium EDTA, and sodium phytate sequentially, stirring until completely dissolved. Then, slowly add hydroxyethyl cellulose and acrylate / C10-30 alkanol acrylate crosspolymer, stirring for 20 minutes and allowing to hydrate for 40 minutes to obtain the hydrated protective phase.

[0177] In a separate oil phase pot, add hydrogenated lecithin, ceramide NP, cholesterol, phytosterols, squalane, C13-15 alkanes, caprylic / capric triglycerides, polyglycerol-10 stearate, and hesperidin methyl chalcone. Heat the oil phase to 72°C and maintain this temperature with stirring for 20 minutes to ensure that hesperidin methyl chalcone is fully dispersed in the lipid phase.

[0178] The oil phase was slowly added to the hydrated protective phase, homogenized at 3000 rpm for 3 minutes, and then homogenized at 5000 rpm for 2 minutes to obtain a liquid crystal layered emulsion system. The system was then cooled by low-speed stirring.

[0179] Separately, take a portion of butylene glycol, a portion of glycerol, and a small amount of deionized water, add caffeine, and stir at 50°C until transparent to obtain a caffeine pre-solution. Once the emulsion system has cooled to below 45°C, add the caffeine pre-solution and stir at low speed for 10 minutes.

[0180] Once the system has cooled to below 35°C, acetyl hexapeptide-8 solution, Bacillus fermentation product, horse chestnut fermentation protective complex, ergothioneine, dipotassium glycyrrhizate, acetylated sodium hyaluronate, sodium hyaluronate, 1,2-hexanediol, and ethylhexylglycerin are added sequentially, and the mixture is stirred at low speed for 15 minutes. The pH is adjusted to 6.4±0.2 using arginine, and the osmotic pressure is monitored and adjusted to 280-320 mOsm / kg. The mixture is then degassed under vacuum, filtered through a 100-mesh filter, and filled into vials to obtain the eye care composition.

[0181] Example 2

[0182] This embodiment provides an eye care product. Compared to Embodiment 1, this embodiment reduces the amount of horse chestnut fermentation protective complex used.

[0183] The formula in this embodiment, expressed as a percentage by weight, is as follows:

[0184] Deionized water, balance; Glycerin 4.50%; Butylene glycol 3.50%; Trehalose I 2.50%; Ectoin I 0.30%; Panthenol 1.00%; Caffeine 0.30%; Acetyl hexapeptide-8 solution 5.00%; Bacillus fermentation product 3.00%; Horse chestnut fermentation protective complex 2.00%; Hesperidin methyl chalcone 0.05%; Ergothioneine 0.10%; Dipotassium glycyrrhizate 0.20%; Acetylated sodium hyaluronate 0.05%; Sodium hyaluronate 0.03%; Hydrogenated lecithin 1.50%; Ceramide NP 0.15%; Cholesterol 0.10%; Phytosterol 0.20%; Squalane 3.00%; C13-15 Alkanes 4.00%; Caprylic / Capric Triglyceride 3.00%; Polyglycerol-10 Stearate 0.60%; Hydroxyethyl Cellulose 0.25%; Acrylic (Ester) / C10-30 Alkyl Acrylate Crosspolymer 0.18%; Disodium EDTA 0.03%; Sodium Phytate 0.02%; 1,2-Hexanediol 0.50%; Ethylhexylglycerin 0.30%; Arginine as needed.

[0185] The preparation method of the horse chestnut fermentation protective complex in this embodiment is the same as that in Example 1. The preparation method of the eye care composition is also the same as that in Example 1, except that the amount of horse chestnut fermentation protective complex added is 2.00%, and deionized water is used to make up to 100%.

[0186] Example 3

[0187] This embodiment provides an eye care product. Compared with Example 1, this embodiment increases the amount of hesperidin methyl chalcone, ceramide NP, and some lipid structural components.

[0188] The formula in this embodiment, expressed as a percentage by weight, is as follows:

[0189] Deionized water, balance; Glycerin 4.50%; Butylene glycol 3.50%; Trehalose I 2.50%; Ectoin I 0.30%; Panthenol 1.00%; Caffeine 0.30%; Acetyl hexapeptide-8 solution 5.00%; Bacillus fermentation product 3.00%; Horse chestnut fermentation protective complex 4.00%; Hesperidin methyl chalcone 0.10%; Ergothioneine 0.10%; Dipotassium glycyrrhizate 0.20%; Acetylated sodium hyaluronate 0.05%; Sodium hyaluronate 0.03%; Hydrogenated lecithin 1.80%; Ceramide NP 0.30%; Cholesterol 0.15%; Phytosterol 0.25%; Squalane 3.00%; C13-15 Alkanes 4.00%; Caprylic / Capric Triglyceride 3.00%; Polyglycerol-10 Stearate 0.70%; Hydroxyethyl Cellulose 0.25%; Acrylic (Ester) / C10-30 Alkyl Acrylate Crosspolymer 0.18%; Disodium EDTA 0.03%; Sodium Phytate 0.02%; 1,2-Hexanediol 0.50%; Ethylhexylglycerin 0.30%; Arginine as needed.

[0190] The preparation method of the horse chestnut fermentation protective complex in this embodiment is the same as that in Example 1.

[0191] The preparation method of the eye care composition is basically the same as that in Example 1, except that 0.10% hesperidin methyl chalcone, 1.80% hydrogenated lecithin, 0.30% ceramide NP, 0.15% cholesterol, 0.25% phytosterols, and 0.70% polyglycerol-10 stearate are added to the oil phase. The oil phase is kept at 72°C and stirred for 25 minutes. The remaining steps are the same as in Example 1.

[0192] Comparative Example 1

[0193] This comparative example does not include the horse chestnut fermentation protection complex.

[0194] The formulation of this comparative example is basically the same as that of Example 1, except that the horse chestnut fermentation protection complex is not added, and the amount is made up to 100% with an equal amount of deionized water.

[0195] The preparation method of this comparative example is the same as that of Example 1, except that the horse chestnut fermentation protection complex is not added during the low-temperature activity addition stage.

[0196] Comparative Example 2

[0197] This comparative example uses ordinary horse chestnut water extract instead of horse chestnut fermentation protection complex to verify the technical effect of horse chestnut fermentation process compared with ordinary water extraction process.

[0198] The preparation method of common horse chestnut water extract is as follows:

[0199] Collect horse chestnut seeds, remove the shells, dry at 45℃ until the moisture content is below 8%, and pulverize to 60 mesh. Weigh 100 g of horse chestnut powder, add 1200 g of deionized water, adjust the pH to 5.6, extract at 45℃ for 1.5 hours, filter, and collect the filtrate. Centrifuge the obtained filtrate at 4000 rpm for 15 minutes, collect the supernatant, and filter through a 0.22 μm filter to obtain the common horse chestnut water extract.

[0200] The eye care composition formulation in this comparative example is basically the same as that in Example 1, except that 4.00% of the horse chestnut fermentation protective complex is replaced with 4.00% of ordinary horse chestnut water extract.

[0201] The preparation method of the eye care product in this comparative example is the same as that in Example 1, except that the common horse chestnut water extract is added during the low-temperature activity addition stage.

[0202] Comparative Example 3

[0203] This comparative example uses common horse chestnut extract instead of horse chestnut fermentation protection complex to verify the effects of the fermentation protection and ultrafiltration fractionation process of this invention on reducing periorbital irritation and system stability.

[0204] The preparation method of common horse chestnut alcohol extract is as follows:

[0205] Collect horse chestnut seeds, remove the shells, dry at 45℃ until the moisture content is below 8%, and pulverize to 60 mesh. Weigh 100 g of horse chestnut powder, add 1200 g of 50% ethanol aqueous solution, extract at 45℃ for 2 hours, and filter. Add another 800 g of 50% ethanol aqueous solution to the residue, extract at 45℃ for 1 hour, filter, and combine the filtrates. Concentrate the combined filtrate under reduced pressure below 45℃ until there is no obvious ethanol odor, add deionized water to make up to the original weight, centrifuge at 4000 rpm for 15 minutes, collect the supernatant, and filter through a 0.22 μm filter to obtain the common horse chestnut ethanol extract.

[0206] The formulation of this comparative example is basically the same as that of Example 1, except that 4.00% of the horse chestnut fermentation protective complex is replaced with 4.00% of ordinary horse chestnut alcohol extract.

[0207] The preparation method of the eye care product in this comparative example is the same as that in Example 1, except that the common horse chestnut extract is added at a low temperature.

[0208] Comparative Example 4

[0209] This comparative example does not include trehalose II during the horse chestnut fermentation process, and is used to verify the effect of trehalose II as a fermentation protectant on the stability and low irritation of horse chestnut ferment.

[0210] The preparation method of the horse chestnut ferment in this comparative example is basically the same as that in Example 1, except that trehalose II is not added during the co-fermentation stage, but 3.0 g of ectoine II is still added. The remaining enzymatic hydrolysis, enzyme inactivation, inoculation, fermentation, ultrafiltration fractionation, and compounding steps are the same as in Example 1. The obtained product is referred to as trehalose II-free horse chestnut ferment.

[0211] The eye care composition formulation of this comparative example is basically the same as that of Example 1, except that 4.00% of the horse chestnut fermentation protective complex is replaced with 4.00% of trehalose-free horse chestnut ferment.

[0212] The preparation method of the eye care product in this comparative example is the same as that in Example 1.

[0213] Comparative Example 5

[0214] This comparative example does not include ectoine II during horse chestnut fermentation to verify the effect of ectoine II as a fermentation protectant on the mildness of the ferment and the compatibility of the eye system.

[0215] The preparation method of the horse chestnut ferment in this comparative example is basically the same as that in Example 1, except that ectoine II is not added during the co-fermentation stage, while 20.0 g of trehalose II is still added. All other conditions are the same as in Example 1. The resulting product is designated as ectoine II-free horse chestnut ferment.

[0216] The formulation of this comparative example is basically the same as that of Example 1, except that 4.00% of the horse chestnut fermentation protective complex is replaced with 4.00% of the ecotoxin II-free horse chestnut ferment.

[0217] The preparation method of the eye care product in this comparative example is the same as that in Example 1.

[0218] Comparative Example 6

[0219] This comparative example omits the ultrafiltration fractionation step of horse chestnut fermentation.

[0220] The preparation method of the horse chestnut ferment in this comparative example is as follows:

[0221] The method described in Example 1 was followed to complete the fermentation of horse chestnut by crushing, compound enzymatic hydrolysis, enzyme inactivation, and co-fermentation with *Lactobacillus plantarum* and *Saccharomyces cerevisiae*, with 20.0 g of trehalose II and 3.0 g of ectoine II added during the fermentation process. After fermentation, the pH was adjusted to 6.0, and the mixture was centrifuged at 4000 rpm for 15 minutes. The supernatant was collected. The obtained supernatant was not subjected to 3000 Da ultrafiltration fractionation but was directly filtered through a 0.22 μm filter to obtain the unfiltered horse chestnut ferment.

[0222] The formulation of this comparative example is basically the same as that of Example 1, except that 4.00% of the horse chestnut fermentation protective complex is replaced with 4.00% of the unfiltered horse chestnut ferment.

[0223] The preparation method of the eye care product in this comparative example is the same as that in Example 1.

[0224] Comparative Example 7

[0225] In this comparative example, hesperidin methyl chalcone was added directly to the aqueous phase without pre-dispersion in the lipid phase.

[0226] The formulation of this comparative example is the same as that of Example 1.

[0227] The preparation methods of this comparative example differ as follows:

[0228] The preparation of the hydrated protective phase was the same as in Example 1. Hydrogenated lecithin, ceramide NP, cholesterol, phytosterols, squalane, C13-15 alkanes, caprylic / capric triglycerides, and polyglycerol-10 stearate were added to the oil phase, but hesperidin methyl chalcone was not added. The oil phase was emulsified with the aqueous phase after being stirred at 72°C for 20 minutes.

[0229] Hesperidin methyl chalcone, in powder form, was added directly to the aqueous phase after the emulsion system was cooled to below 45°C, and stirred at low speed for 20 minutes. The remaining steps for adding caffeine pre-solution, adding low-temperature activity, adjusting pH, and controlling osmotic pressure were the same as in Example 1.

[0230] Comparative Example 8

[0231] This comparative example uses a one-time mixing process.

[0232] The formulation of this comparative example is the same as that of Example 1.

[0233] The preparation method for this comparative example is as follows:

[0234] Add deionized water, glycerin, butylene glycol, trehalose I, ectoine I, panthenol, disodium EDTA, sodium phytate, hydroxyethyl cellulose, acrylate / C10-30 alkanol acrylate crosspolymer, caffeine, acetyl hexapeptide-8 solution, Bacillus fermentation product, horse chestnut fermentation protective complex, ergothioneine, dipotassium glycyrrhizate, acetylated sodium hyaluronate, sodium hyaluronate, 1,2-hexanediol, and ethylhexylglycerin to the mixing bowl and stir to combine.

[0235] Separately, hydrogenated lecithin, ceramide NP, cholesterol, phytosterols, squalane, C13-15 alkanes, caprylic / capric triglycerides, polyglycerol-10 stearate, and hesperidin methyl chalcone were mixed and heated to 72°C, then added to the mixing bowl and homogenized at 5000 rpm for 5 minutes. The mixture was then cooled, the pH was adjusted to 6.4±0.2 using arginine, and the osmotic pressure was adjusted to 280-320 mOsm / kg. After degassing and filtration, the mixture was filled into containers.

[0236] Comparative Example 9

[0237] This comparative example omits trehalose I and ectoine I from the eye care composition, but retains trehalose II and ectoine II derived from the fermentation process in the horse chestnut fermentation protective complex, to verify the role of the formulation-end hydration protection system of the composition.

[0238] The formulation of this comparative example is basically the same as that of Example 1, except that trehalose I 2.50% and ectoine I 0.30% are not added separately, and the amount is made up to 100% with an equal volume of deionized water. The horse chestnut fermentation protective complex is still prepared according to the method of Example 1, and it still contains trehalose II and ectoine II derived from the fermentation process.

[0239] The preparation method of this comparative example is basically the same as that of Example 1, except that trehalose I and ectoine I are not added to the hydrated protective phase.

[0240] Freeze-thaw stability testing

[0241] 1. Test Objective

[0242] The evaluation examples and comparative examples were assessed to determine whether stratification, precipitation, crystallization, flocculation, or emulsification of the system occurred when peptides, fermentation products, flavonoids, saponins, and antioxidants were present.

[0243] 2. Testing Methods

[0244] Take 50 g of each of the samples from Examples 1-3 and Comparative Examples 1-9 and place them in transparent centrifuge tubes. Observe the initial appearance, including color, odor, uniformity, presence of visible particles, oil-water separation, or flocculent matter.

[0245] Then, take 30 g of sample and place it in a 50 mL centrifuge tube. Centrifuge at 4000 rpm for 30 min at 25℃. After centrifugation, let it stand for 10 min and observe whether the sample shows oil-water separation, floating oil layer, bottom sedimentation, flocculation, coarse particles, or obvious crystallization.

[0246] 3. Judgment Criteria

[0247] If the sample shows no stratification, no precipitation, and no visible crystal points after centrifugation, it is recorded as "stable".

[0248] If the sample shows slight turbidity or a small amount of particles, but no obvious stratification, it is recorded as "basically stable".

[0249] If a sample shows obvious stratification, precipitation, or crystal points, it is marked as "unstable".

[0250] High temperature storage stability testing

[0251] 1. Test Objective

[0252] The study evaluated the stability of the emulsion structure, the risk of flavonoid crystallization, and the risk of caffeine recrystallization of the samples under alternating low and high temperature conditions.

[0253] 2. Testing Methods

[0254] Take 50 g of each of the samples from Examples 1-3 and Comparative Examples 1-9 and put them into sealed glass bottles.

[0255] The conditions for each freeze-thaw cycle are as follows: -5℃ for 12 h; -40℃ for 12 h. The above process constitutes one cycle, and a total of 5 cycles are performed.

[0256] After each cycle, observe the appearance of the sample. After the 5th cycle, restore the sample to 25°C, let it stand for 24 hours, and then observe its appearance, fluidity, whether it separates into layers, whether crystal points appear, whether flocculation occurs, and color changes.

[0257] 3. Detection Indicators

[0258] Record the following indicators: 1. Appearance uniformity; 2. Whether it separates into layers; 3. Whether flavonoid crystals appear; 4. Whether caffeine crystals appear; 5. Whether turbidity or precipitation occurs; 6. Viscosity change rate.

[0259] The viscosity change rate is calculated using the following formula:

[0260] Viscosity change rate (%) = |Vt - V0| / V0 * 100%

[0261] Where V0 is the viscosity before freeze-thaw and Vt is the viscosity after freeze-thaw.

[0262] 4. Judgment Criteria

[0263] If the viscosity change rate is ≤10% and there is no stratification or obvious crystal points, it is judged to have good freeze-thaw stability.

[0264] The viscosity change rate was 10%-20%, and only slight turbidity was observed, indicating that the freeze-thaw stability was average.

[0265] If the viscosity change rate is greater than 20%, or if obvious stratification, crystal points, or precipitation occurs, it is judged as having poor freeze-thaw stability.

[0266] Low temperature crystallization testing

[0267] 1. Test Objective

[0268] The compatibility of active ingredients, color stability, emulsification stability, and odor changes of the samples under accelerated conditions were evaluated.

[0269] 2. Testing Methods

[0270] Take 100 g of each of the samples from Examples 1-3 and Comparative Examples 1-9, put them into sealed glass bottles, and store them in a constant temperature incubator at 45°C for 30 days.

[0271] Samples were taken and observed on day 0, day 7, day 14, and day 30.

[0272] 3. Detection Indicators

[0273] The testing items include:

[0274] 1. Appearance; 2. Color; 3. Odor; 4. pH; 5. Viscosity; 6. Crystallization; 7. Layering; 8. Acetyl hexapeptide-8 retention rate; 9. Ergothioneine retention rate.

[0275] 4. pH test

[0276] Take 5.0 g of sample, add 45.0 g of deionized water, stir well to prepare a 10% aqueous dispersion. Measure the pH value using a calibrated pH meter. Each sample was tested in triplicate, and the average value was taken.

[0277] 5. Viscosity test

[0278] The viscosity of the sample was determined using a rotational viscometer. The test conditions were as follows: rotor: No. 4 rotor; rotation speed: 12 rpm; test temperature: 25℃; equilibration time: 30 s.

[0279] Each sample was tested in parallel three times, and the average value was taken.

[0280] Hesperidin methyl chalcone retention testing

[0281] 1. Test Objective

[0282] To evaluate the crystallization risk of caffeine and hesperidin methyl chalcone under low-temperature storage conditions.

[0283] 2. Testing Methods

[0284] Take 30 g of each sample from Examples 1-3 and Comparative Examples 1-9, place them in transparent glass bottles, and store them in a refrigerator at 4°C for 30 days. Observe the samples on day 0, day 7, day 14, and day 30.

[0285] Observation methods include:

[0286] 1. Observation with the naked eye; 2. Observation with a 10x magnifying glass; 3. Observation with a polarizing microscope.

[0287] The observation method using a polarizing microscope is as follows:

[0288] Take approximately 0.05 g of sample, place it on a glass slide, cover it with a coverslip, and gently press it into a thin layer. Observe the number and morphology of crystals under a polarizing microscope. Observe five fields of view for each sample randomly.

[0289] 3. Evaluation of Crystal Quantity

[0290] Evaluation based on the number of crystals visible in each field of view:

[0291] Level 0: No crystals observed; Level 1: 1-5 crystals / field of view; Level 2: 6-20 crystals / field of view; Level 3: >20 crystals / field of view; Level 4: Abundant crystals or platy crystals.

[0292] Caffeine retention and risk of crystallization testing

[0293] 1. Test Objective

[0294] To evaluate the stability of hesperidin methyl chalcone in different formulation structures, especially to compare the differences between Examples 1-3 and Comparative Example 7.

[0295] 2. Sample processing

[0296] Accurately weigh 1.00 g of sample, add 20 mL of methanol, and extract by sonication for 30 min. After cooling to room temperature, bring the volume to 25 mL with methanol. Take the supernatant and filter it through a 0.22 μm organic filter membrane as the test solution.

[0297] 3. HPLC testing conditions

[0298] Chromatographic column: C18 column, 250 mm × 4.6 mm, 5 μm; mobile phase: acetonitrile-0.1% phosphoric acid aqueous solution; gradient elution: acetonitrile 30%-70%, 20 min; flow rate: 1.0 mL / min; column temperature: 30℃; detection wavelength: 280 nm; injection volume: 10 μL.

[0299] 4. Calculation Formula

[0300] Retention rate of hesperidin methyl chalcone (%) = Ct / C0 * 100%

[0301] Wherein, C0 is the content of hesperidin methyl chalcone in the sample on day 0, and Ct is the content of hesperidin methyl chalcone in the sample after storage.

[0302] Aescin source active compatibility testing

[0303] 1. Test Objective

[0304] To evaluate the solubility stability of caffeine in samples and the risk of recrystallization after low-temperature storage.

[0305] 2. Sample processing

[0306] Accurately weigh 0.50 g of sample, add 25 mL of 50% methanol aqueous solution, extract by sonication for 20 min, cool and centrifuge, take the supernatant and filter it through a 0.22 μm filter membrane as the test solution.

[0307] 3. HPLC testing conditions

[0308] Chromatographic column: C18 column, 250 mm × 4.6 mm, 5 μm; mobile phase: methanol-water = 30:70; flow rate: 1.0 mL / min; detection wavelength: 273 nm; column temperature: 30℃; injection volume: 10 μL.

[0309] 4. Calculation Formula

[0310] Caffeine retention rate (%) = Ct / C0 * 100%

[0311] Where C0 is the caffeine content on day 0, and Ct is the caffeine content after storage.

[0312] In vitro eye irritation surrogate assessment

[0313] 1. Test Objective

[0314] To evaluate the compatibility of the horse chestnut fermentation protective complex with conventional water extract, alcohol extract, trehalose II-free fermentation product, ectoine II-free fermentation product, and non-ultrafiltration fermentation product in ocular systems.

[0315] 2. Testing Methods

[0316] Take 30 g of each of the samples from Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Comparative Example 5 and Comparative Example 6, store them at 45℃ for 30 days, and centrifuge them at 4000 rpm for 30 min simultaneously.

[0317] Observe and record:

[0318] 1. Appearance; 2. Whether it is cloudy; 3. Whether there is sediment; 4. Whether there is a change in odor; 5. Change in viscosity; 6. Whether there is an increase in foaming;

[0319] 7. Whether there are stratifications after centrifugation.

[0320] 3. Judgment Criteria

[0321] If the viscosity change rate of the sample is ≤10% after high-temperature storage and there is no stratification or obvious precipitation after centrifugation, it is considered to have good compatibility.

[0322] If the viscosity change rate is 10%-20%, or there is slight turbidity, it is considered to have average compatibility.

[0323] If the viscosity change rate is greater than 20%, or if there is obvious stratification, precipitation, or increased foaming, it is considered to have poor compatibility.

[0324] Human ocular use assessment

[0325] 1. Test Objective

[0326] To evaluate the potential irritation of the sample when used in the periorbital area.

[0327] 2. Red blood cell hemolysis test

[0328] Fresh rabbit or sheep red blood cells were washed three times with PBS buffer to prepare a 2% red blood cell suspension. The sample was diluted to a 1.0% concentration with PBS. 1 mL of the diluted sample was mixed with 1 mL of the 2% red blood cell suspension and incubated at 37°C for 30 min. After incubation, the sample was centrifuged at 3000 rpm for 5 min, and the supernatant was collected. The absorbance was measured at 540 nm. PBS was used as the negative control, and 0.1% sodium dodecyl sulfate solution was used as the positive control.

[0329] 3. Calculation Formula

[0330] Hemolysis rate (%) = (As - An) / (Ap - An) * 100%

[0331] Where As is the absorbance of the sample group, An is the absorbance of the negative control, and Ap is the absorbance of the positive control.

[0332] 4. Judgment Criteria

[0333] A hemolysis rate of ≤5% is considered low irritation.

[0334] The hemolysis rate is 5%-10%, which is considered a mild risk of irritation.

[0335] A hemolysis rate greater than 10% indicates a high level of irritation.

[0336] Moisture retention test

[0337] 1. Test Objective

[0338] The moisture retention and water holding capacity of the sample after the formation of a hydrated film were evaluated.

[0339] 2. Testing Methods

[0340] Take 0.50 g of sample, spread it evenly in a glass petri dish with a diameter of 5 cm, place it in an environment of 25℃ and 50% relative humidity for 30 min to equilibrate, weigh it, and record it as W0.

[0341] The sample was then placed in a dry environment at 25°C and 30% relative humidity, and weighed at 2 h, 4 h, and 8 h, respectively, and recorded as Wt.

[0342] 3. Calculation Formula

[0343] Moisture retention rate (%) = Wt / W0 * 100%

[0344] Where W0 is the initial mass and Wt is the mass at the corresponding time point.

[0345] Figures 1-3

[0346] 1. Test Objective

[0347] The evaluation focused on the sample's effectiveness in improving eye bags, dark circles, eye fatigue, fine lines around the eyes, and user comfort.

[0348] 2. Subjects

[0349] Thirty healthy participants, aged 35-60, with mild to moderate under-eye bags, dark circles, or signs of eye fatigue, will be recruited. Participants will not use other eye care products during the testing period.

[0350] 3. How to use

[0351] A randomized controlled trial was conducted, using either half-face or single-eye approaches. Each subject used the test sample around one eye and a blank matrix or control sample on the other. The sample was applied twice daily, morning and evening, at a dose of approximately 0.03 g each time, for 28 consecutive days.

[0352] 4. Test Time Points

[0353] Tests were conducted on day 0, day 7, day 14, and day 28.

[0354] 5. Eye bag volume test

[0355] A three-dimensional skin imaging device was used to acquire images of the area under the eyes, and the volume or degree of protrusion of the eye bags was analyzed.

[0356] The improvement rate of eye bags is calculated using the following formula:

[0357] Improvement rate of eye bags (%) = (B0 - Bt) / B0 * 100%

[0358] Where B0 is the volume of the eye bags on day 0, and Bt is the volume of the eye bags at the corresponding time point after use.

[0359] 6. Dark Circle Test

[0360] L in the subocular region was measured using a spectrophotometer. * a * b * The value is calculated, and ITA° is determined.

[0361] ITA°=tan -1(L * -50) / b * *180 / π

[0362] An increased ITA° indicates a reduction in dark circles around the eyes and a brighter complexion.

[0363] 7. Periocular fatigue score

[0364] The scores are given by trained evaluators using a 5-point scale:

[0365] 1 point: No obvious fatigue state; 2 points: Mild fatigue state; 3 points: Moderate fatigue state; 4 points: Obvious fatigue state; 5 points: Severe fatigue state.

[0366] The fatigue improvement rate is calculated using the following formula:

[0367] Fatigue improvement rate (%) = (S0 - St) / S0 * 100%

[0368] Where S0 is the rating on day 0, and St is the rating after use.

[0369] 8. Eye area fine lines test

[0370] The depth of fine lines at the corners of the eyes was measured using a rapid 3D skin imaging system or a silicone replication method.

[0371] The improvement rate of fine lines is calculated using the following formula:

[0372] Fine line improvement rate (%) = (D0 - Dt / D0 * 100%)

[0373] Where (D0) is the wrinkle depth on day 0, and Dt is the wrinkle depth after use.

[0374] 9. User comfort evaluation

[0375] On day 28, participants completed a questionnaire and evaluated the following items: 1. Stinging; 2. Eye irritation; 3. Redness; 4. Stickiness; 5. Refreshing sensation; 6. Soothing sensation around the eyes; 7. Reduction in puffiness around the eyes.

[0376] Each item is rated on a scale of 0 to 5, where 0 is considered "none" and 5 is considered "very obvious".

[0377] Test Results and Analysis

[0378] Table 1 Results of appearance, centrifugation, freeze-thaw and high-temperature stability

[0379]

[0380] As shown in Table 1, Examples 1-3 maintained good appearance stability under centrifugation, freeze-thaw cycles, high and low temperature conditions. Among them, Example 1 showed the best overall stability. Comparative Example 7, due to the absence of hesperidin methyl chalcone in the lipid phase, exhibited obvious crystal formation after low-temperature storage. Comparative Example 8, due to the use of a one-time mixing process, experienced strong heat treatment and shearing of caffeine, peptides, fermentation products, and ergothioneine, resulting in stratification, discoloration, and crystal points. Comparative Example 9 lacked the formulation-terminal trehalose I and ectoine I, leading to a decrease in the system's hydration protection capacity, manifested as viscosity changes and slight water separation.

[0381] Table 2 Retention rate of main active ingredients after storage at 45℃ for 30 days

[0382]

[0383] Table 3. Compatibility and irritation results of components derived from horse chestnut.

[0384]

[0385] As shown in Table 3, the horse chestnut fermentation protection complex in Example 1 exhibited the lowest periorbital irritation, indicating that the combined enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, trehalose II / ectoine II fermentation protection, and ultrafiltration fractionation can reduce irritating small molecules and interfacial active impurities in the horse chestnut-derived active ingredients. Comparative Examples 3 and 6 showed higher irritation, suggesting that ordinary ethanol extracts and unfiltered fermentation broths are not suitable as preferred periorbital systems.

[0386] Table 4 Results of in vitro water retention rate

[0387]

[0388] As shown in Table 4, Examples 1-3 exhibited good water-holding capacity. In Comparative Example 9, after removing trehalose I and ectoine I from the formulation, the water retention rate decreased significantly after 8 hours, indicating that trehalose I and ectoine I not only contribute to the stability of the active ingredient but also to the formation of the periorbital hydration film and water retention.

[0389] Table 5. Results of periorbital appearance improvement after 28 days of continuous use.

[0390]

[0391] Note: The stinging rating is based on a scale of 0 to 5, with lower values ​​indicating less stimulation.

[0392] As shown in Table 5, Example 1 demonstrated the best performance in terms of improvement rate of eye bags, improvement rate of fatigue state, and overall comfort; Example 3 was slightly better in terms of ITA° improvement and fine line improvement, indicating that enhancing the flavonoid-barrier lipid phase is beneficial for improving dark circles and barrier-related fine lines. In Comparative Example 1, the lack of the horse chestnut fermentation protective complex significantly reduced the improvement rate of eye bags. Comparative Example 8, using a one-time mixing process, showed the lowest human evaluation results, indicating that decreased stability of the active ingredient directly affects the improvement effect on the appearance of the periocular area.

[0393] The test results above show that Examples 1-3 of the present invention are superior to the comparative examples in terms of stability, retention rate of active ingredients, low irritation, inflammation regulation, microcirculation regulation, glycation inhibition, barrier strengthening, and improvement of the human periorbital area.

[0394] Example 1, as the preferred embodiment, showed the most balanced performance across all indicators, indicating a good synergistic effect among the horse chestnut fermentation protection complex, flavonoid lipid phase, caffeine pre-dissolving system, low-temperature partitioning process, and the formulation-end trehalose I / ectoine I hydration protection system.

[0395] Comparative Examples 2 and 3 show that ordinary Aesculus hippocastanum water or alcohol extracts cannot be equivalent to replacing the Aesculus hippocastanum fermentation protective complex. Comparative Examples 4 and 5 show that the addition of trehalose II and ectoine II during fermentation helps improve the mildness and system compatibility of the Aesculus hippocastanum fermentation product. Comparative Example 6 shows that ultrafiltration fractionation can reduce irritating small molecules and interfacial active impurities. Comparative Example 7 shows that hesperidin methyl chalcone needs to be pre-dispersed in the lipid phase to reduce the risk of crystallization. Comparative Example 8 shows that a one-time mixing process significantly reduces the retention rate of heat-sensitive active ingredients and affects stability. Comparative Example 9 shows that trehalose II and ectoine II in the Aesculus hippocastanum fermentation protective complex cannot completely replace trehalose I and ectoine I at the formulation end, the latter contributing independently to the osmotic pressure, hydration state, and active ingredient stability of the entire ocular composition.

[0396] Antera 3D Three-Dimensional Crow's Feet Evaluation Test

[0397] Experimental Objective

[0398] The effects of the eye care composition of Example 1 of the present invention on improving crow's feet, fine lines and skin texture around the eyes were evaluated.

[0399] Subjects

[0400] Thirty healthy female subjects, aged 35 to 60 years, were selected.

[0401] Inclusion criteria: (1) Visible crow's feet and fine lines at the corners of the eyes; (2) No serious inflammation or skin diseases around the eyes; (3) No medical aesthetic wrinkle removal treatment in the past month; (4) No use of other anti-wrinkle eye creams or eye serums during the trial period.

[0402] Test Sample Usage Instructions

[0403] Subjects used the eye care composition prepared in Example 1 once in the morning and once in the evening each time. Each time, about 0.10g of the sample was taken and evenly applied to the outer corner of both eyes and the lower eyelid area. The sample was gently massaged until fully absorbed and used continuously for 28 days.

[0404] Test environment

[0405] The test was conducted in a constant temperature and humidity laboratory: temperature: 22±2℃; relative humidity: 50±5%; subjects sat quietly for 30 minutes before the test and were prohibited from using any eye care products for 8 hours before the test.

[0406] Testing instruments

[0407] Delfin Antera3D® 3D Skin Analysis System.

[0408] This instrument uses multispectral imaging technology and a 3D reconstruction algorithm to obtain 3D morphological information of the skin surface and automatically identify wrinkle areas, calculating:

[0409] (1) Total area of ​​wrinkles (mm) 2 (2) Total volume of wrinkles (mm) 3 (3) Average wrinkle depth (μm) (4) Maximum wrinkle depth (μm) (5) Total wrinkle length (mm) (6) Skin roughness parameter Ra (7) Skin texture uniformity index

[0410] Test time point

[0411] T0: Before use; T14: 14 days after use; T28: 28 days after use.

[0412] Images of the corner of the eye region were acquired under the same lighting, shooting angle, and positioning conditions.

[0413] Result Calculation

[0414] The wrinkle improvement rate (%) is calculated using the following formula:

[0415] Improvement rate (%) = (A0 - At) / A0 * 100

[0416] Where: A0 is the parameter value before use; At is the parameter value at the corresponding time point.

[0417] Table 6 Results of Antera3D Three-Dimensional Crow's Feet Evaluation Experiment

[0418]

[0419] Among them, the appendix ​The images show Antera 3D images (from left to right: 0d, 14d, 28d) of changes in crow's feet before (0d), after (14d), and after (28d) use of the product in Example 1 for subjects A to C. The wrinkle area improvement rates for the three subjects were 57.3%, 76.8%, and 64.1%, respectively, consistent with the overall statistical results.

[0420] The Antera3D three-dimensional skin analysis system was used to continuously detect the eye area of ​​the subjects. The results showed that after 14 days of continuous use of the eye care composition obtained in Example 1 of this invention, the number and length of fine lines at the corners of the eyes were significantly reduced; after 28 days of continuous use, the wrinkle area, wrinkle volume, and total wrinkle length all decreased further.

[0421] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions 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. An eye care composition, characterized in that, The composition comprises acetyl hexapeptide-8, Bacillus fermentation product, hesperidin methyl chalcone, ergothioneine, ectoine, and caffeine.

2. The eye care composition according to claim 1, characterized in that, It also contains horse chestnut ferment.

3. The eye care composition according to claim 2, characterized in that, The preparation method of the horse chestnut ferment includes the following steps: (1) Crush the horse chestnut raw material and add it to water; (2) Add cellulase and pectinase for compound enzymatic hydrolysis; (3) After enzyme inactivation, inoculate with Lactobacillus plantarum and Saccharomyces cerevisiae for synergistic fermentation; (4) Add trehalose and ectoine during fermentation; (5) After fermentation, perform ultrafiltration fractionation; (6) Collect the permeate and mix it with part of the retentate to obtain horse chestnut ferment.

4. An eye care product, characterized in that, It comprises the eye care composition of claim 1.

5. The eye care product according to claim 4, characterized in that, The eye care composition comprises the following components by weight percentage: Trehalose: 0.5–5.0%; Ectoin: 0.05–1.0%; Glycerin: 1.0–10.0%; Butylene glycol: 1.0–10.0%; Caffeine: 0.05–1.0%; Acetyl hexapeptide-8: 0.1–10.0%; Bacillus fermentation product: 0.1–10.0%; Horse chestnut ferment: 0.1–10.0%; Hesperidin methyl chalcone: 0.001–1.0%; Ergothioneine: 0.001–1.0%; Dipotassium glycyrrhizate: 0.01–1.0%; Acetylated sodium hyaluronate: 0.001–1.0%; Sodium hyaluronate: 0.001–1.0%; Hydrogenated lecithin: 0.1–5.0%; Ceramide NP: 0.001–1.0%; Cholesterol: 0.001–1.0%; Phytosterols: 0.001–2.0%; Squalane: 0.1–10.0%; C13-15 alkanes: 0.1–10.0%; Caprylic / Capric triglycerides: 0.1–10.0%; Polyglycerol fatty acid ester emulsifier: 0.05–5.0%; Hydroxyethyl cellulose: 0.05–2.0%; Acrylic (ester) crosspolymer / C10-30 alkanol acrylate crosspolymer: 0.01–2.0%; Metal ion complexing agent: 0.001–0.5%; Preservative system: 0.1–3.0%; Deionized water: balance.

6. A method for preparing the eye care product according to claim 5, characterized in that, Includes the following steps: (1) Add trehalose, ectoine, glycerol, butanediol and metal ion complexing agent to the aqueous phase and mix and dissolve them, then add to the thickening system for hydration; (2) Hesperidin methyl chalcone, hydrogenated lecithin, cholesterol, ceramide and oil components are heated together; (3) Add the flavonoid lipid-encapsulated phase obtained in step (2) to step (1); (4) Caffeine is pre-dissolved in a trehalose-polyol pre-dissolved system and then added to the system obtained in step (3); (5) When the system temperature drops to below 35-40℃, add acetyl hexapeptide-8, Bacillus fermentation product, horse chestnut fermentation product and ergothioneine; (6) Adjust the pH of the system to 6.2-6.8 and control the osmotic pressure to 280-320 mOsm / kg to obtain the eye care composition; The horse chestnut ferment is prepared from horse chestnut raw material through compound enzymatic hydrolysis, lactic acid bacteria-yeast co-fermentation, and ultrafiltration fractionation.

7. The eye care composition according to any one of claims 1 to 3, characterized in that: The composition is used to improve one or more of the following: eye bags, dark circles, eyelid sagging, eye fatigue, and sleep deprivation-induced periorbital aging.

8. The use of the eye care composition according to any one of claims 1 to 3 in the preparation of an eye care product for regulating periorbital inflammation, microcirculation, lymphatic integrity and barrier stability.

9. The application according to claim 8, characterized in that: The regulation includes one or more of the following: IL-33 / ST2 inflammatory pathway regulation, VEGF / VE-Cadherin permeability regulation, Emilin-1 / Integrin α9β1 lymphoid integrity regulation, AGEs / ALEs glycation regulation, and desmosome and tight junction enhancement.

10. The application according to claim 8, characterized in that: The eye care product is one or more of the following: eye cream, eye serum, eye mask, liquid crystal emulsion, gel, and microcapsule sustained-release formulation.

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