Compositions, liposomes thereof and use in the manufacture of skin care products

Abstract

The present application relates to the cosmetic technical field, especially to a composition, a liposome thereof and application in preparing skin care products, wherein NAD, ergothioneine, boswelia, ectoine and glutathione are combined, the obtained composition can have good moisturizing, skin elasticity improving, redness removing and repairing effects, and has synergistic effects in moisturizing, skin elasticity improving and other aspects. The preparation of the liposome improves the stability and permeability of the composition, so that it can play a better effect when used as a skin care product.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and more particularly to compositions and their liposomes and their use in the preparation of skin care products. Background Technology

[0002] In today's society, people are paying increasing attention to their personal image and skin health, leading to more diverse and sophisticated skincare needs. Consumers are not only seeking basic moisturizing but also desiring significant results in areas such as anti-aging, whitening, and repairing the skin barrier. To meet these growing skincare demands, the market offers a wider variety of skincare products, from traditional lotions and creams to emerging products like serums, ampoules, and masks, with product forms constantly being innovated.

[0003] Despite the diversity of product forms, most skincare products fall short of expectations in actual use. Consumers report that after using certain anti-aging products for a period of time, the reduction in wrinkles and skin firmness are not significant; whitening products show slow whitening effects, failing to achieve the desired results; and products designed to improve the skin barrier have limited effectiveness in addressing skin sensitivity and dryness. The main reasons for this are twofold: firstly, the selection of inappropriate active ingredients leads to insufficient activity; secondly, the product formulation design is not optimal, limiting stability and penetration. Specifically:

[0004] Regarding physiological activity, while existing skincare products utilize a variety of active ingredients, they do not adequately consider the synergistic effects between these components. Different skincare ingredients have varying mechanisms of action and functional characteristics. Without a scientifically sound combination and synergistic design, the components cannot form a complementary system; instead, they may interfere with or cancel each other out, preventing the active ingredients from fully realizing their skincare efficacy.

[0005] Regarding stability and permeability, many active ingredients are prone to oxidation and degradation during product storage, leading to a decrease in the content of effective ingredients and affecting product efficacy. Meanwhile, the stratum corneum, as a natural barrier, hinders the absorption of most active ingredients, making it difficult for a large number of effective ingredients to penetrate deep into the skin to exert their effects. Simultaneously improving the stability and permeability of skincare products presents numerous contradictions. Measures to improve stability, such as changing the formulation system or adding stabilizers, may alter the product's physicochemical properties, reducing the permeability of active ingredients; conversely, methods to enhance permeability may compromise product stability and accelerate the degradation of active ingredients.

[0006] Therefore, how to significantly improve the skin penetration rate of active ingredients while ensuring product stability, thereby enhancing the overall efficacy of skincare products, has become an important issue that urgently needs to be addressed in the current skincare field, and requires further in-depth research and exploration. Summary of the Invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a composition and its liposomes and their use in the preparation of skin care products.

[0008] The composition provided by this invention consists of NAD+, glutathione, ergothioneine, bosine, and ectoine.

[0009] In this invention, the mass ratio of NAD+, glutathione, ergothioneine, bosine, and ectoine is (1.5–2.5):(0.8–1.2):(0.3–0.4):(0.8–1.2):(0.8–1.2).

[0010] In a specific embodiment, the mass ratio of NAD+, glutathione, ergothioneine, bosine, and ectoine is 2:1:0.5:1:1.

[0011] This invention combines NAD+, glutathione, ergothioneine, pro-xylene, and ectoine. NAD+ possesses multiple benefits, including anti-aging, energy provision, and cell repair promotion. Ergothioneine exhibits strong antioxidant properties and can increase NAD+ levels in cells by inhibiting NADPH and NADH oxidases. It also participates in GSH / GSSG redox coupling, inhibiting glutathione peroxidase, increasing GSH concentration, and strengthening the antioxidant network system. The antioxidant properties of ergothioneine and glutathione can be applied to lipids in liposomes to reduce putrefaction and oxidation. Ectoine provides excellent hydration, anti-allergy, and anti-inflammatory effects, and enhances cell repair capabilities. Pro-xylene promotes the synthesis of glycosaminoglycans and collagen, resulting in excellent anti-wrinkle and firming effects. This invention combines the aforementioned components to form a composition with superior effects in moisturizing, anti-aging, and repairing redness, particularly exhibiting a synergistic effect in reducing redness, improving skin elasticity, and moisturizing.

[0012] The present invention also provides a liposome prepared from excipients and the composition as described above.

[0013] Although the composition provided by this invention has good physiological activity, each component is difficult to penetrate the stratum corneum directly into the skin, and NAD+ is unstable, while glutathione and ergothioneine are easily oxidized, so their efficacy is easily affected during use. Encapsulating them with liposomes can effectively improve the penetration rate and stability.

[0014] In this invention, the excipients include phospholipids, cholesterol, surfactants, and solvents.

[0015] In some embodiments, the phospholipid includes, but is not limited to, natural phospholipids such as PC50, PC60, PC70, and PC80, and synthetic phospholipids such as hydrogenated lecithin, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylethanolamine, 1,2-distearate-sn-propanetriyl-3-phosphocholine, and PEG-phospholipids. Preferably, the phospholipid is phospholipid PC-60.

[0016] In some embodiments, the cholesterol may be replaced by other similar components, such as at least one of lanosterol, sitosterol, stigmasterol, and ergosterol, with cholesterol being preferred.

[0017] In some embodiments, the surfactant is at least one selected from Tween 80, Tween 20, Span 80, Span 20, PEG-7 hydrogenated castor oil, PEG-40 hydrogenated castor oil, PEG-20 almond glyceryl ester (arachidic acid), PEG-20 glyceryl triisostearate, PEG-6 caprylic / capric glyceryl ester, PEG-7 glyceryl cocoate, glyceryl isostearate, polyglycerol-3 polyricinoleate, polyglycerol-3 diisostearate, polyglycerol-2 isostearate, polyglycerol-2 triisostearate, polyglycerol-2 tetraisostearate, propylene glycol isostearate, and polyglycerol-4 laurate. Preferably, the surfactant is Tween-80.

[0018] In some embodiments, the solvent is ethanol and water.

[0019] In some embodiments, the excipients further include preservatives, including but not limited to at least one selected from phenoxyethanol, sorbic acid, caprylyl glycol, 1,2-hexanediol, and p-hydroxyacetophenone. Preferably, the preservative is phenoxyethanol.

[0020] In this embodiment of the invention, the liposomes comprise the following raw materials in parts by weight: 0.8-1.2 parts phospholipids, 0.2-0.3 parts cholesterol, 1.5-2.5 parts NAD+, 0.8-1.2 parts glutathione, 0.4-0.6 parts ergothioneine, 0.8-1.2 parts boswellicine, 0.8-1.2 parts ectoine, 0.1-0.2 parts Tween, 0.2-0.4 parts NaH2PO4, and 0.2 parts preservative;

[0021] In some specific embodiments, the liposomes comprise the following raw materials in parts by weight: 1 part phospholipid, 0.25 parts cholesterol, 2 parts NAD+, 1 part glutathione, 0.5 parts ergothioneine, 1 part bosine, 1 part ectoine, 0.15 parts Tween, 0.3 parts NaH2PO4, and 0.2 parts preservative.

[0022] In this embodiment of the invention, the raw materials for preparing the liposomes also include ethanol and water, and the volume ratio of ethanol to water is (10-20):50, preferably 15:50.

[0023] Furthermore, the present invention also provides a method for preparing liposomes as described above, comprising:

[0024] Phospholipids and cholesterol were dissolved in ethanol to obtain the oil phase;

[0025] The aqueous phase was prepared by dissolving NAD+, glutathione, ergothioneine, bosine, ectoine, Tween, and preservative in an aqueous solution of NaH2PO4.

[0026] After preheating the oil phase and the aqueous phase to 50-70°C, the aqueous phase is poured into the oil phase and stirred to obtain a mixed solution;

[0027] After removing ethanol from the mixed solution, it is homogenized 3 to 4 times under a pressure of 900 to 1000 bar to obtain a liposome solution.

[0028] In some embodiments, the mass-to-volume ratio of phospholipids to ethanol in the oil phase is 1 g:(10-20) mL, preferably 1 g:15 mL;

[0029] In some embodiments, the concentration of NaH2PO4 in the aqueous phase is (0.5–0.7) g / 100 mL, and the concentration of NAD+ is (3–5) g / 100 mL. Preferably, the concentration of NaH2PO4 is 0.6 g / 100 mL, and the concentration of NAD+ is 4 g / 100 mL.

[0030] Preferably, the oil phase and water phase of the present invention are mixed after being preheated to 60°C.

[0031] Furthermore, the present invention also provides the use of the composition as described above, the liposomes described above, or the liposomes prepared by the method described above in the preparation of skin care products.

[0032] In this invention, the skincare product is prepared by forming a complex liposome from NAD+, ergothioneine, pro-xylene, ectoine, and glutathione. Compared to non-liposome forms, it has better stability and permeability; compared to liposomes containing a single active ingredient, it has richer efficacy. Skincare products prepared from this complex can exert stronger anti-aging, wrinkle-reducing, whitening, and skin barrier-improving effects, achieving a closed loop of "defense-repair-regeneration" in skincare and meeting various skincare needs.

[0033] Furthermore, the present invention also provides a skin care product comprising a cosmetic matrix and the composition as described above, the liposomes described above, or the liposomes prepared by the method described above.

[0034] In this invention, the dosage form of the skincare product is at least one of the following: toner, lotion, essence, emulsion, face cream, serum, ampoule, eye cream, eye serum, face mask (sheet, cream, mud mask, gel mask), facial cleanser, facial cleanser, facial foam, scrub, exfoliating gel, lotion, face cream, sunscreen, makeup base, lotion, body lotion, hand cream, lip balm, lip mask, hand cream, neck cream, BB cream, CC cream, BB cream, essential oil, skin care oil, spray, freeze-dried powder, gel, jelly, mud paste, ointment, sleeping mask, sheet mask, eye mask, nose mask, and leave-on mask.

[0035] As a feasibility example, the skincare product described is a skincare serum. Its excipients include water, glycerin, carbomer, 1,2-hexanediol, pH adjuster, and preservatives.

[0036] Preferably, the liposomes in the skin care product or the liposomes prepared by the method have a mass fraction of 20%.

[0037] Preferably, the skin care product has a pH value of 4.5 to 5.5. More preferably, the skin care product has a pH value of 5.0.

[0038] Preferably, the pH adjuster is triethanolamine and the preservative is phenoxyethanol.

[0039] Furthermore, the present invention also provides a skin care method comprising applying the skin care product as described above to the skin surface.

[0040] In this invention, the application methods include, but are not limited to, smearing, patting, massaging, applying wet compresses, applying sheets (such as sheet masks), dotting (such as local acne treatment products), spraying (such as sprays), wiping (such as applying toner with a cotton pad), applying thickly (such as applying a thick layer of a mask or face cream), mixing (such as layering or blending different products), rolling (such as rollerball serums), pressing (such as gently pressing with the palm of the hand to promote absorption), and applying in circular motions (such as cleansing or face cream).

[0041] This invention combines NAD, ergothioneine, pro-xylene, ectoine, and glutathione to create a composition that exhibits excellent moisturizing, skin elasticity-improving, redness-reducing, and repairing effects, with synergistic effects in multiple aspects such as moisturizing and improving skin elasticity. Preparing it as a liposome improves the stability and permeability of the composition, thereby enhancing its effectiveness when used as a skincare product. Attached Figure Description

[0042] Figure 1 From left to right: Liposome solutions prepared in Examples 1-4;

[0043] Figure 2The liposome solutions prepared in Example 1 (left) and Comparative Example 1 (right) are shown after being placed at 40°C for 30 days. Detailed Implementation

[0044] This invention provides compositions and their liposomes, and their application in the preparation of skincare products. Those skilled in the art can refer to this document and appropriately modify the process parameters to achieve the desired results. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in this invention. The methods and applications of this invention have been described through preferred embodiments. Those skilled in the art will clearly be able to modify or appropriately alter and combine the methods and applications described herein without departing from the content, spirit, and scope of this invention to realize and apply the technology of this invention.

[0045] Unless otherwise defined in this invention, the scientific and technical terms associated with this invention shall have the meanings understood by one of ordinary skill in the art.

[0046] The terms “comprising,” “including,” and “having” are used interchangeably to indicate the inclusiveness of a scheme, meaning that the scheme may contain elements other than those listed. It should also be understood that the use of “comprising,” “including,” and “having” herein also provides for schemes “consisting of…”.

[0047] The term "and / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can mean: A exists alone, A and B exist simultaneously, or B exists alone. A and B can be singular or plural.

[0048] In this application, "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refer to any combination of these items, including any combination of single or multiple items.

[0049] In this invention, nicotinamide adenine dinucleotide (NAD+), also known as coenzyme I, is an important intracellular coenzyme involved in energy metabolism and various biochemical reactions. With age, the level of NAD+ in the body gradually declines, a phenomenon closely related to aging and various diseases. Supplementing with NAD+ can promote cell repair and regeneration, reduce wrinkles, delay aging, and brighten and whiten the skin.

[0050] In this invention, glutathione is a tripeptide composed of glutamic acid, cysteine, and glycine. It is the most abundant non-protein thiol compound in cells and exists in two forms in vivo: GSH (reduced form) and GSSG (oxidized form). GSH / GSSG redox coupling is an important antioxidant mechanism in the human body. GSH plays a major role in the body and can directly neutralize free radicals (ROS). Increasing GSH levels can effectively enhance antioxidant effects.

[0051] In this invention, ergothioneine (EGT) is a natural antioxidant existing in two isomers. Its powerful antioxidant capacity stems from the balance of thiol tautomerism. In vivo, EGT typically exists primarily in the thione state, ready to exert its antioxidant effect by capturing free radicals, and is subsequently reduced to the thiol state. EGT exhibits strong antioxidant capacity even at extremely low concentrations, capable of scavenging various reactive oxygen species, nitrogen species, and free radical products.

[0052] In this invention, Pro-Xylane is a glycoprotein mixture derived from xylose, possessing multiple functions including anti-wrinkle effects, whitening, and improving the skin barrier. The extracellular matrix of human cells in the dermis contains a large amount of glycosaminoglycans, which have strong water absorption and viscosity, helping to maintain the space of the extracellular matrix in the dermis. Based on biomimetic principles, Pro-Xylane promotes glycosaminoglycan synthesis, thereby achieving a dermal repair effect.

[0053] In this invention, ectoine is a natural amino acid derivative with amphiphilic and zwitterionic characteristics. It can accumulate in large quantities in cells to resist high osmotic pressure shocks. It can also act as a stabilizer for biological macromolecules and the whole cell, as well as a stress protectant. It can help the skin relieve various external pressures (dry environment, surfactant damage, ultraviolet radiation, etc.), enhance cellular immunity, help the skin resist the invasion of microorganisms and allergens, and play an anti-allergic and anti-inflammatory role. In addition, it can also increase cell repair ability and accelerate the recovery of skin health.

[0054] The numerical ranges and parameters involved in this invention have been presented as precisely as possible in the specific embodiments. However, any numerical value inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise explicitly stated, it should be understood that all numerical ranges or specific data used in this disclosure may have a reasonable deviation within a certain range, such as ±10%, ±5%, ±1%, or ±0.5%.

[0055] The embodiments and comparative examples of this invention describe some examples. These embodiments illustrate certain implementations of the invention. However, this does not mean that the effects of the invention can only be achieved in these examples. In fact, any concentration of each component between the two endpoints shown in the embodiments can achieve good effects such as anti-aging, wrinkle reduction, whitening, and improvement of the skin barrier.

[0056] The test materials used in this invention are all commercially available products. It should be understood that in the various embodiments of this application, the sequence numbers of the above processes do not imply a specific order of execution. Some or all steps can be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of this application. The invention is further illustrated below with reference to embodiments:

[0057] Example 1

[0058] The components and their contents are shown in Table 1:

[0059] Table 1

[0060]

[0061] (1) Weigh out the phospholipids and cholesterol according to the amounts in Table 1 and add them to a beaker. Add ethanol and stir at 60°C until completely dissolved to obtain the oil phase.

[0062] (2) Weigh the phosphate according to the amount in Table 1 and add it to the beaker. Dissolve it with pure water by stirring. Then add NAD+, glutathione, ergothioneine, bosine, ectoine, Tween 80 and phenoxyethanol and stir until completely dissolved to obtain the aqueous phase.

[0063] (3) Both the oil phase and the water phase are preheated at 60°C and stirred. The water phase is quickly added to the oil phase and stirred at 60°C for 1 hour to obtain a mixed solution.

[0064] (4) Transfer the mixed solution to a rotary evaporator, remove the ethanol at 50°C, and then transfer it to a homogenizer. Homogenize it 3 to 4 times at 900 to 1000 bar to obtain the liposome solution.

[0065] Example 2

[0066] (1) The dissolution of the oil phase and the aqueous phase is the same as in Example 1;

[0067] (2) The oil phase was transferred to a round-bottom flask and evaporated under reduced pressure at 50°C to remove ethanol, resulting in an oil film that was uniformly attached to the inner wall of the round-bottom flask.

[0068] (3) Preheat the aqueous phase to 45°C. Place the eggplant-shaped flask with the oil film in a 45°C water bath and slowly add the aqueous phase while stirring until the oil film on the wall is completely dissolved in the aqueous phase to form a homogeneous solution. Then transfer it to a homogenizer and homogenize it 3 to 4 times at a pressure of 900 to 1000 bar to obtain the liposome solution.

[0069] Example 3

[0070] (1) The dissolution of the oil phase and the aqueous phase is the same as in Example 1;

[0071] (2) Both the oil phase and the water phase are preheated at 60°C and stirred. The oil phase is quickly added to the water phase and stirred at 60°C for 1 hour to obtain a mixed solution.

[0072] (3) Transfer the mixed solution to a rotary evaporator, remove the ethanol at 50°C, and then transfer it to a homogenizer. Homogenize it 3 to 4 times at a pressure of 900 to 1000 bar to obtain a liposome solution.

[0073] Example 4

[0074] (1) The dissolution of the oil phase and the aqueous phase is the same as in Example 1;

[0075] (2) Both the oil phase and the water phase are preheated at 60°C and stirred. The water phase is quickly added to the oil phase and stirred at 60°C for 1 hour to obtain a mixed solution.

[0076] (3) Transfer the mixed solution to a round-bottom flask, rotate it under reduced pressure at 50°C to remove ethanol and water, forming a thin film on the inner wall of the round-bottom flask. Then add an aqueous solution containing phosphate and stir at 45°C for 1 hour to obtain a mixed solution. Transfer the mixed solution to a homogenizer and homogenize it 3-4 times at a pressure of 900-1000 bar to obtain a liposome solution.

[0077] Example 5

[0078] (1) The oil phase and aqueous phase were dissolved in the same way as in Example 1, and the aqueous solutions were adjusted to pH 4.0, 5.0, 6.0 and 7.0 respectively;

[0079] (2) Both the oil phase and the water phase are preheated at 60°C and stirred. The oil phase is quickly added to the water phase and stirred at 60°C for 1 hour to obtain a mixed solution.

[0080] (3) Transfer the mixed solution to a rotary evaporator, remove the ethanol at 50°C, and then transfer it to a homogenizer. Homogenize it 3 to 4 times at a pressure of 900 to 1000 bar to obtain a liposome solution.

[0081] Comparative Examples 1-9

[0082] The components and contents of Comparative Examples 1–9 are shown in Table 2:

[0083] Table 2

[0084]

[0085] Liposomes were prepared according to the method in Example 1, with the following specific steps:

[0086] (1) Weigh out the phospholipids and cholesterol according to the amounts in Table 2 and add them to a beaker. Add ethanol and stir at 60°C until completely dissolved to obtain the oil phase.

[0087] (2) Weigh the phosphate according to the dosage in Table 2 and add it to the beaker. Dissolve it with pure water by stirring. Then add NAD+, Tween 80, phenoxyethanol, glutathione or ergothioneine or bosine or ectoine, stir, and adjust the pH to 5.0 after complete dissolution to obtain the aqueous phase.

[0088] (3) Both the oil phase and the water phase are preheated at 60°C and stirred. The oil phase is quickly added to the water phase and stirred at 60°C for 1 hour to obtain a mixed solution.

[0089] (4) Transfer the mixed solution to a rotary evaporator, remove the ethanol at 50°C, and then transfer it to a homogenizer. Homogenize it 3 to 4 times at 900 to 1000 bar to obtain the liposome solution.

[0090] Comparative Example 10 Aqueous Solution

[0091] According to the dosage in Table 3, weigh the phosphate and add it to a beaker. Dissolve it with pure water by stirring. Then add NAD+, Tween 80, phenoxyethanol, glutathione, ergothioneine, bosine, and ectoine. Stir until completely dissolved and adjust the pH to 5.0 to obtain an effective aqueous solution (not a liposome solution).

[0092] Table 3. Composition ratios of Comparative Example 10

[0093]

[0094] Efficacy testing

[0095] 1. Encapsulation efficiency determination

[0096] The encapsulation efficiency of liposomes was determined using dialysis. 1 mL of the liposome suspension from Examples 1-4 was transferred to a dialysis bag (molecular weight cutoff 3.5 kD), suspended in 100 mL of phosphate buffer (pH 6.0, 0.05 M), and dialyzed for 12 h with slow stirring. The levels of NAD, glutathione, ergothioneine, bosine, and ectoine in the dialysis fluid were measured and recorded as the free active components of the liposomes. The encapsulation efficiency was calculated using the formula.

[0097] Encapsulation efficiency = (1 - in vitro free active ingredient / total active ingredient in liposome solution) × 100%

[0098] The encapsulation efficiency test results are shown in Table 4:

[0099] Table 4 Encapsulation efficiency of active ingredients prepared by different methods

[0100] Encapsulation efficiency (%) NAD+ Glutathione Ergothioneine Ikdoin Bosine Example 1 80 82 81 76 85 Example 2 50 54 51 46 52 Example 3 62 55 49 60 51 Example 4 79 80 75 69 72

[0101] The results show that the preparation method has a certain impact on the encapsulation efficiency, with Example 1 having the highest encapsulation efficiency, followed by Example 4.

[0102] 2. Liposome peroxide value detection

[0103] The peroxide value of phospholipids in liposome solutions was determined according to GB 5009.227-2023, using the indicator titration method.

[0104] In Examples 1-4, after being placed at 40°C for 60 days, no phospholipid oxidation was detected, and no off-odor was produced. In Comparative Examples 1-8, the phospholipids began to oxidize within 7 days, producing a sour odor. After 30 days, they were almost completely oxidized, reaching the oxidation termination period. Figure 2 This indicates that the combination of multiple active substances has a better antioxidant effect on liposomes, while the antioxidant performance of active substances used alone is poor.

[0105] Table 5. Detection of liposome peroxidation value

[0106]

[0107]

[0108] "-" indicates that the peroxide value could not be measured due to complete oxidation.

[0109] 3. Stability Comparison

[0110] In Example 5, liposomes were prepared under different pH conditions, and their stability was measured at 25°C. Liposomes prepared under aqueous phase conditions of pH 4.0–6.0 were more stable. pH affects both the stability of the active ingredient and the stability of the liposomes; pH 5.0 is recommended as the optimal condition.

[0111] Table 6. Detection of liposome peroxidation value

[0112] Overall retention rate (%) initial 7 days 14 days 30 days pH 4.0 100 102 105 98 pH 5.0 100 104 100 99 pH 6.0 100 98 96 98 pH 7.0 100 96 94 75

[0113] Considering both encapsulation efficiency and stability, the preparation method in Example 1 is superior. The methods of Example 1 and other comparative examples are used to verify the efficacy of the active ingredients.

[0114] Example 6

[0115] The essence was prepared according to Table 7 using liposomes from Examples 1 and Comparative Examples 6-10 as the active ingredient.

[0116] Table 7. Liposome solutions used in the preparation of serums

[0117]

[0118] The test products were formulated from Example 1 and Comparative Examples 6-10 through application examples.

[0119] A group of participants used the test product, and the product's moisturizing, firming, and soothing effects were comprehensively evaluated using instrumental testing. The participants were aged 28 to 55 years, with an average age of 46 years.

[0120] The experimental results were compared using statistical methods. The data in Tables 8-11 show that the essence with Example 1 as the active ingredient had the most significant effects in moisturizing, firming, and soothing. In Control Examples 6-8, the lack of active ingredients resulted in poor efficacy. In Control Example 9, although the content of active ingredients was consistent, its efficacy was lower than that of the liposome form when applied in an aqueous solution.

[0121] Table 8 Efficacy Evaluation Scheme

[0122]

[0123] Table 9 Results of Skin Stratum Corneum Moisture Content Test

[0124]

[0125] Table 10 Results of the Skin Elasticity Efficacy Test

[0126]

[0127] Table 11 Results of Skin Tightness Test

[0128]

[0129] Table 12 Results of Skin Redness Test

[0130]

[0131] According to the Bliss independent model, the sample of Example 1 was used as the combined sample, and Comparative Example 7 and Comparative Example 8 were two single drugs. The CI value was calculated based on the effect data relative to Example 9 (the test sample data minus the effect data of Example 9).

[0132] Calculations showed that the CI values ​​were all greater than 1 in terms of the improvement rates of skin stratum corneum moisture, skin elasticity, skin firmness, and skin redness, demonstrating a good synergistic effect.

[0133] Example 7

[0134] 1. Experimental objective:

[0135] The study investigated the subcutaneous permeation, intradermal retention, and epidermal residue of the complex liposomes in transdermal absorption experiments.

[0136] 2. Experimental Principle:

[0137] 1. GB / T 27818-2011 In vitro test methods for skin absorption of chemicals

[0138] 2.FDA In Vitro Permeation Test Studies for Topical Drug ProductsSubmitted inANDAs

[0139] 3. Experimental Design:

[0140] The excised skin was fixed on the Franz diffusion apparatus with the stratum corneum facing up and the dermis facing down in contact with the receiving medium in the receiving cell.

[0141] The sample was dropped onto the stratum corneum, ensuring close contact with the skin, with 3 replicates for each sample;

[0142] Set the water bath temperature to 32℃ and the magnetic stir bar speed to 300rpm to ensure that there are no air bubbles in the water bath jacket;

[0143] At 2 and 6 hours, 1 ml of sample was taken from the receiving cell using a sampling tube. After each sampling, the same volume of fresh receiving solution was added and air bubbles were removed.

[0144] After the 6-hour experiment, samples were collected from the skin surface and the inner skin, and analyzed by HPLC with the receiving solution at each time point.

[0145] 4. Experimental Materials

[0146] 4.1 Instrument Information:

[0147] Table 13. Information on instruments for the skin penetration test

[0148] name factory model Transdermal diffusion device Jingtuo Instruments TP-6 High Performance Liquid Chromatography Thermo Fisher VanquishCore Electronic balance Sartorius Secura225D-1CN pH meter Mettler FE28

[0149] 4.2 Sample Information:

[0150] Liposomes of Comparative Example 10 and Example 1.

[0151] 5. Experimental Procedure:

[0152] 5.1 Receiving medium: Weigh 8.0g NaCl, 0.2g KCl, 1.44g Na2HPO4, and 0.24g KH2PO4 and dissolve them in 800mL distilled water. Adjust the solution to 7.4 with HCl, and finally add distilled water to make up to 1L.

[0153] 5.2 Test solution: Accurately measure 0.5 mL each of Example 1 and Comparative Example 10, place them in 10 mL volumetric flasks, add receiving medium to dissolve and dilute to the mark, shake well, and the solution is ready;

[0154] 5.3 Reference solutions: Transfer the test solution and dilute it to prepare solutions containing approximately 1, 10, 100, 500, and 2000 μg per mL, respectively, as reference solutions for the standard curve;

[0155] 5.4 Take Bama miniature pig skin, cut it to a suitable size, and soak it in the receiving medium for about 10 minutes. Add an appropriate amount of receiving medium to the diffusion cup and place it in the diffusion cell at a constant temperature. Take the soaked pig skin, wipe it dry, place it on the diffusion cup and fix it, then add receiving medium until there are no air bubbles in the water bath jacket, ensuring that the receiving medium is in full contact with the lower surface of the skin. Transfer 300 μL of the test solution and add it dropwise into the supply chamber, setting the rotation speed to 300 rpm. Take 1 mL samples at 2 h and 6 h and replenish the solution accordingly.

[0156] 5.5 After the experiment, use a dropper to remove the residual solution from the supply chamber and clean the supply chamber multiple times with the receiving medium, taking 1 mL of the receiving medium each time, repeating 3 times. Transfer the residual solution and cleaning solution to a 10 mL volumetric flask, add the receiving medium to dilute to the mark, and shake well to obtain the residual solution on the skin.

[0157] 5.6 Remove the pigskin, cut it into pieces, place it in a 10mL volumetric flask, add an appropriate amount of receiving medium, sonicate for 30 minutes, then dilute to the mark with receiving medium and shake well to obtain the intradermal retention solution.

[0158] 6. Data Results:

[0159] In Comparative Example 10, no permeation was detected in the transdermal experiment. The active ingredient in Example 1 had a relatively high transdermal permeability, with intradermal retention 1.5 to 10 times that of Comparative Example 10. This indicates that liposomes can improve the transdermal permeability of the active ingredient.

[0160] Table 14. Data from the skin penetration test (6h)

[0161]

[0162] The above are merely preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A composition comprising NAD+, glutathione, ergothioneine, bosine and ectoine.

2. The composition according to claim 1, characterized in that, The mass ratio of NAD+, glutathione, ergothioneine, bosine, and ectoine is (1.5–2.5): (0.8–1.2): (0.3–0.4): (0.8–1.2): (0.8–1.2).

3. Liposomes, which are prepared from excipients and the composition according to claim 1 or 2; The excipients include phospholipids, cholesterol, surfactants, and solvents.

4. The liposomes according to claim 3, characterized in that, The surfactant is Tween, and the solvent is ethanol and water.

5. The liposomes according to claim 4, characterized in that, It comprises the following raw materials in parts by weight: 0.8-1.2 parts phospholipids, 0.2-0.3 parts cholesterol, 1.5-2.5 parts NAD+, 0.8-1.2 parts glutathione, 0.4-0.6 parts ergothioneine, 0.8-1.2 parts boswelliae, 0.8-1.2 parts ectoine, 0.1-0.2 parts Tween, 0.2-0.4 parts NaH2PO4, and 0.2 parts preservative; It also includes ethanol and water, wherein the volume ratio of ethanol to water is (10-20):

50.

6. A method for preparing liposomes according to any one of claims 3 to 5, comprising: Phospholipids and cholesterol were dissolved in ethanol to obtain the oil phase; The aqueous phase was prepared by dissolving NAD+, glutathione, ergothioneine, bosine, ectoine, Tween, and preservative in an aqueous solution of NaH2PO4. After preheating the oil phase and the aqueous phase to 50-70°C, the aqueous phase is poured into the oil phase and stirred to obtain a mixed solution; After removing ethanol from the mixed solution, it is homogenized 3 to 4 times under a pressure of 900 to 1000 bar to obtain a liposome solution.

7. The preparation method according to claim 6, characterized in that, In the oil phase, the mass-to-volume ratio of phospholipids to ethanol is 1 g: (10-20) mL; In the aqueous phase, the concentration of NaH2PO4 is (0.5-0.7) g / 100 mL, and the concentration of NAD+ is (3-5) g / 100 mL.

8. The use of the composition according to claim 1 or 2, the liposomes according to any one of claims 3 to 5, or the liposomes prepared by the method according to claim 6 or 7 in the preparation of skin care products.

9. A skin care product comprising a cosmetic base and the composition of claim 1 or 2, the liposomes of any one of claims 3 to 5, or the liposomes prepared by the method of claim 6 or 7.

10. A skin care method comprising applying the skin care product of claim 9 to the skin surface.

Images