Epiphyllum extract composition based on circadian rhythm gene regulation and preparation method thereof

Abstract

The invention relates to the technical field of combination of circadian rhythm regulation and skin aging resistance, in particular to a broad-leaved epiphyllum extract composition based on circadian rhythm gene regulation and a preparation method of the broad-leaved epiphyllum extract composition. The raw materials comprise 8-15% of a broad-leaved epiphyllum stem cell rhythm regulation extract, 1-3% of melatonin targeted delivery microspheres, 2-5% of an NAD + precursor compound and the like; the extract is subjected to supercritical extraction and inclusion with cyclodextrin, and the microspheres contain RGD peptide for targeted delivery; the composition also contains a photoresponse transdermal factor, a bionic sebum repair system and the like, and the anti-aging effect is improved by synergistically regulating and controlling the circadian rhythm gene through multiple components. According to the composition, through the cooperation of the broad-leaved epiphyllum extract and multiple components, the circadian rhythm gene is accurately regulated and controlled, the NAD < + > / SIRT1 pathway is activated, and collagen synthesis is enhanced; the innovative delivery system improves component absorption, repairs skin barriers, has the characteristics of high efficiency and mildness, and is suitable for anti-aging care of multiple skins.

Description

Technical Field

[0001] The present invention relates to the technical field of combining circadian rhythm regulation with skin anti-aging, and specifically to an epiphyllum extract composition based on circadian rhythm gene regulation and a preparation method thereof. Background Art

[0002] With the accelerated pace of modern life, circadian rhythm disorders have become a common health problem worldwide. They not only affect physiological metabolism, but also accelerate the aging process of the skin. As the largest organ in the human body, the aging manifestations of the skin are closely related to the circadian rhythm. Abnormal expression of core clock genes (such as BMAL1 and PER2) will lead to reduced synthesis of type I collagen and enhanced oxidative stress response, which in turn will cause wrinkles, sagging and other aging phenomena. Traditional anti-aging cosmetics focus more on antioxidant or moisturizing functions, lack of targeted design for circadian rhythm regulation, and it is difficult to fundamentally improve skin aging problems caused by rhythm disorders.

[0003] The molecular mechanism of circadian rhythm depends on the transcription-translation negative feedback loop, in which BMAL1 and CLOCK serve as key transcriptional activators, and their expression levels directly affect the regulation of downstream aging-related genes. Studies have shown that melatonin can improve circadian rhythm by regulating pineal gland secretion, but exogenous melatonin has poor stability and low transdermal efficiency, and it is difficult for a single ingredient to achieve multi-pathway synergistic anti-aging effects. In addition, there is cross-regulation between the NAD+ / SIRT1 signaling pathway and the circadian rhythm. A decrease in NAD+ levels will lead to a decrease in SIRT1 activity and accelerate cell aging, but the delivery efficiency and bioavailability of existing NAD+ precursors (such as nicotinamide) are insufficient, which limits their application effect.

[0004] As an important model organism, zebrafish has a high homology between its core circadian clock genes and humans, and its embryos are transparent and have a short developmental cycle, which facilitates the real-time observation of gene expression changes. Existing studies have confirmed that quercetin in Epiphyllum extract has antioxidant and anti-inflammatory effects, but its potential in regulating circadian rhythm genes has not yet been fully developed. In addition, the transdermal absorption efficiency of active ingredients in traditional cosmetic formulas is generally low, especially large molecules (such as polysaccharides and peptides) are difficult to penetrate the skin barrier, resulting in insufficient effective concentration. Therefore, the development of an anti-aging cosmetic composition and its preparation method that can accurately regulate circadian rhythm genes, enhance the synergistic effect of multiple ingredients, and have efficient transdermal ability has become the key to solving the bottleneck of existing technologies.

[0005] (1) Technical problems solved

[0006] In response to the deficiencies of the prior art, the present invention provides an Epiphyllum oxypetalum extract composition based on circadian rhythm gene regulation and a preparation method thereof.

[0007] (2) Technical solution

[0008] A composition of an epiphyllum extract based on circadian rhythm gene regulation; the raw materials include, by weight percentage:

[0009] Epiphyllum stem cell rhythm regulation extract: produced through supercritical CO2 extraction combined with β-cyclodextrin inclusion technology, containing quercetin, isorhamnetin and circadian rhythm regulating peptide precursors, which form a host-guest complex after β-cyclodextrin inclusion, with an inclusion rate of ≥85%;

[0010] Melatonin targeted delivery microspheres: Melatonin and RGD tripeptide are encapsulated by chitosan, with a particle size of 80-120 nm. The core-shell structure is formed by electrostatic adsorption, and the surface RGD peptide density is ≥5×10 9 molecules / μm2, RGD tripeptide is arginine-glycine-aspartic acid;

[0011] NAD + Precursor complex: Nicotinamide mononucleotide and pterostilbene combine through hydrogen bonds to form a stable complex with a molecular weight of 635.6Da, which can synergistically increase intracellular NAD + level;

[0012] Telomere-protecting polysaccharide: Fucoidan extracted from deep-sea red algae, with a sulfate content ≥ 25%, can specifically bind to the telomerase reverse transcriptase subunit;

[0013] Photoresponsive transdermal factor: A conjugate of ethyl 4-dimethylaminocinnamate and cholesterol sulfate, which can form transient pores under 365nm ultraviolet light, promoting the penetration of active ingredients; the photoresponsive transdermal factor coupling reaction formula is:

[0014]

[0015] Bionic sebum repair system: contains ceramide NP, cholesterol, and fatty acids in a molar ratio of 1:1:1, simulating the lipid structure of the skin barrier;

[0016] Antioxidant synergist: Resveratrol and ferulic acid are connected by an ester bond to form a bisphenol antioxidant molecule, which increases the efficiency of scavenging free radicals by 3 times;

[0017] Sustained-release preservative: a compound of caprylhydroxamic acid and glyceryl caprylate, which is slowly released through liposome encapsulation;

[0018] The balance is deionized water and a pH regulator, wherein the pH regulator is citric acid or triethanolamine, and the pH is adjusted to 5.5-6.5.

[0019] Preferably, the Epiphyllum stem cell rhythm regulation extract also contains gamma-aminobutyric acid, namely GABA, which can enhance the expression of the circadian rhythm gene BMAL1 by activating GABAA receptors.

[0020] Preferably, the melatonin targeted delivery microspheres are prepared by the following method: dissolving melatonin and RGD tripeptide in ethanol-water, adding chitosan solution, adding sodium tripolyphosphate solution dropwise under ultrasonic conditions, cross-linking to form microspheres, collecting by centrifugation and then freeze-drying.

[0021] Preferably, the NAD + In the precursor complex, the molar ratio of nicotinamide mononucleotide to pterostilbene is 2:1, and it is prepared into an amorphous powder by freeze-drying, and the particle size after reconstitution is ≤200nm.

[0022] Preferably, the added amount of the light-responsive transdermal factor is 0.5%.

[0023] Preferably, the purity of ceramide NP in the bionic sebum repair system is ≥98%, and the fatty acid is a mixture of oleic acid and linoleic acid in a ratio of 2:1, which can form a bionic lipid film with a thickness of 50-80 nm on the skin surface.

[0024] Preferably, the antioxidant activity of the antioxidant synergist is ≥90%, and the activity retention rate after storage at 45° C. for 3 months is ≥85%.

[0025] Preferably, the method for preparing the Epiphyllum cyrtonema extract composition based on circadian rhythm gene regulation according to any of the above items comprises the following steps:

[0026] S1: Active ingredient pretreatment: Epiphyllum stem cell rhythm regulation extract and β-cyclodextrin were mixed in a ratio of 1:5 and stirred at 50°C for 4 hours to form a nanoinclusion complex;

[0027] S2: Microsphere preparation: Melatonin targeted delivery microspheres were prepared according to the method of claim 3 and dispersed in deionized water to form a stable suspension;

[0028] S3: Oil phase preparation: Dissolve the biomimetic sebum repair system components and light-responsive transdermal factors in caprylic / capric triglyceride and stir evenly;

[0029] S4: Aqueous phase preparation: NAD + The precursor complex, telomere-protecting polysaccharide, and antioxidant synergist are dissolved in deionized water, and a slow-release preservative is added;

[0030] S5: Emulsification: Slowly add the oil phase to the water phase and emulsify in a homogenizer for 10 minutes to form an oil-in-water emulsion;

[0031] S6: Microsphere loading: Cool to 35-40°C, add melatonin targeted delivery microsphere suspension and Epiphyllum nanoinclusion complex, and stir for 30 minutes;

[0032] S7: pH adjustment and filling: Adjust the pH to 5.8-6.2 with citric acid, filter through a 0.45μm filter membrane, and aseptically fill in a light-proof container.

[0033] Preferably, 0.1-0.3% of γ-aminobutyric acid is added in step S6, and ultrasound-assisted dispersion is used for 15 minutes.

[0034] Preferably, the prepared composition needs to be irradiated with 365 nm ultraviolet light for 3-5 minutes before use to activate the light-responsive transdermal factor and form a transient permeation channel.

[0035] (3) Beneficial technical effects

[0036] Compared with the existing technology, the beneficial effects of the present invention are:

[0037] 1. Accurately regulate circadian rhythm genes to fundamentally delay skin aging

[0038] The Epiphyllum stem cell rhythm-regulating extract in the composition is rich in quercetin, isorhamnetin, and gamma-aminobutyric acid (GABA), which can reshape the rhythmic network through the following pathways: Activating core clock genes: Quercetin stimulates the BMAL1 promoter region, significantly increasing BMAL1 mRNA expression and restoring circadian rhythm amplitude; GABA activates GABAA receptors, enhancing heterodimer formation between BMAL1 and CLOCK protein, synergistically promoting the transcription of downstream anti-aging genes. Optimizing the rhythmic negative feedback loop: Isorhamnetin can significantly reduce the overexpression of the PER2 gene, alleviating oxidative stress-induced circadian rhythm disorders, while simultaneously upregulating CRY1a gene expression and enhancing rhythm stability.

[0039] 2. Multiple ingredients work synergistically to fully activate anti-aging pathways. The complex formed by NMN and pterostilbene significantly increases intracellular NAD+ levels, activates SIRT1 deacetylase activity, promotes type I collagen synthesis, significantly increases mRNA expression, and improves skin elasticity. Fucoidan specifically binds to telomerase, extending telomere length and delaying cellular aging. The biomimetic sebum repair system forms a biomimetic lipid film on the skin's surface, repairing barrier function, significantly reducing transepidermal water loss, and enhancing skin's moisturizing ability.

[0040] 3. An innovative delivery system improves the bioavailability of active ingredients. Chitosan-RGD microspheres significantly increase melatonin accumulation in the basal layer of the skin through electrostatic adsorption and RGD peptide-mediated cellular endocytosis, while also preventing photodegradation and significantly enhancing stability. The ethyl 4-dimethylaminocinnamate-cholesterol sulfate conjugate forms transient pores under 365nm UV light, significantly enhancing the transdermal efficiency of active ingredients such as quercetin and NMN, addressing the absorption challenge of large molecules.

[0041] 4. Safe and practical, the composition's pH is adjusted to 5.5-6.5, closely matching the skin's natural acid-base environment. The sustained-release preservative dosage is ≤0.5%, resulting in lower irritation than traditional preservative systems. Zebrafish embryo toxicity studies have demonstrated a high survival rate at a concentration of 40 mg / L, and human patch testing has confirmed its non-sensitizing properties, making it suitable for anti-aging treatments for all skin types. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 The present invention is a flow chart for preparing an epiphyllum extract composition based on circadian rhythm gene regulation;

[0043] Figure 2 1 is a bar graph of BMAL1 mRNA increase rate and PER2 mRNA decrease rate in the examples and comparative examples;

[0044] Figure 3 It is a line graph of the NAD+ content increase rate and the COLLAGENⅠ mRNA increase rate of the embodiment and the comparative example;

[0045] Figure 4 is a bar graph comparing the reduction rate of initial transepidermal water loss and transdermal efficiency of the examples and comparative examples;

[0046] Figure 5 This is the nuclear magnetic resonance hydrogen spectrum of the light-responsive transdermal factor proposed in the present invention. DETAILED DESCRIPTION

[0047] according to Figures 1 to 5 , the specific implementation methods of the present invention are as follows:

[0048] Example 1

[0049] Raw material formula (weight percentage):

[0050] Epiphyllum stem cell rhythm regulation extract: 12% (containing quercetin 14.2%, γ-aminobutyric acid 0.2%) Melatonin targeted delivery microspheres: 2% (chitosan embedded, RGD peptide density 6×10 9 molecules / μm2)

[0051] NAD + Precursor complex: 3.5% (nicotinamide mononucleotide: pterostilbene = 2:1)

[0052] Fucoidan: 1.2% (molecular weight 50-100kDa, sulfate content ≥25%)

[0053] Photoresponsive transdermal factor: 0.5% (4-dimethylaminocinnamic acid ethyl ester-cholesterol sulfate conjugate) Bionic sebum repair system: 13% (ceramide NP 3%, cholesterol 4%, fatty acid 6%)

[0054] Antioxidant Booster: 2% (Resveratrol Ferulate)

[0055] Slow-release preservative: 0.5% (caprylhydroxamic acid 0.3%, glyceryl caprylate 0.2%)

[0056] Balance: deionized water and pH adjuster (citric acid)

[0057] Preparation process:

[0058] (1) Active ingredient pretreatment:

[0059] Take 120g of Epiphyllum extract and 600g of β-cyclodextrin, add ethanol-water (1:4) mixed solvent, stir magnetically at 50°C for 4 hours, remove the solvent by rotary evaporation and freeze-dry to obtain a nano inclusion complex (inclusion rate 88%).

[0060] (2) Microsphere preparation:

[0061] Melatonin (20 g) and RGD tripeptide (10 g) were dissolved in 1000 mL of ethanol-water (1:1) solution, 2000 mL of 2% chitosan solution (pH 5.5) was added, and 500 mL of 1% sodium tripolyphosphate solution was added dropwise under 40 kHz ultrasonic conditions. After cross-linking for 30 minutes, the mixture was centrifuged (10,000 rpm, 15 min) and freeze-dried to obtain microspheres with a particle size of 95 nm.

[0062] (3) Oil phase preparation:

[0063] 30 g of ceramide NP, 40 g of cholesterol, 60 g of fatty acid and 5 g of light response factor were dissolved in 500 mL of caprylic / capric triglyceride and stirred at 75° C. until completely dissolved.

[0064] (4) Preparation of aqueous phase:

[0065] Dissolve 50 g of nicotinamide mononucleotide and 25 g of pterostilbene in 3000 mL of deionized water, add 12 g of fucoidan, 20 g of antioxidant synergist and preservative, and stir at 55°C until clear.

[0066] (5) Emulsification process:

[0067] The oil phase was slowly poured into the water phase, and homogenized and emulsified at 10,000 rpm for 10 minutes to form a milky white emulsion with a particle size of 200-300 nm.

[0068] (6) Microsphere loading:

[0069] The temperature was lowered to 38°C, 200 mL of microsphere suspension and Epiphyllum nanoinclusion complex were added, ultrasonic dispersion was performed at 30 kHz for 15 minutes, and then stirred for 30 minutes.

[0070] (7) Finished product preparation:

[0071] Adjust the pH to 5.8 with citric acid, filter through a 0.45 μm filter membrane, and aseptically fill in brown glass bottles.

[0072] Performance test results:

[0073] Circadian rhythm regulation: BMAL1 gene expression increased by 52% (qRT-PCR detection)

[0074] Improved cellular metabolism: NAD + Content increased by 68% (HPLC)

[0075] Barrier function: Transepidermal water loss reduced by 38% (measured by TEWL instrument)

[0076] Transdermal absorption: Quercetin transdermal efficiency reaches 55% (Franz diffusion cell method)

[0077] Safety: 0 allergic reactions in 30 patch tests

[0078] Example 2

[0079] Raw material formula (weight percentage):

[0080] Epiphyllum stem cell rhythm regulation extract: 8% (containing quercetin 12%, gamma-aminobutyric acid 0.1%)

[0081] Melatonin targeted delivery microspheres: 1% (chitosan embedded, RGD peptide density 3×10 9 molecules / μm2)

[0082] NAD + Precursor complex: 2% (nicotinamide mononucleotide: pterostilbene = 2:1)

[0083] Fucoidan: 0.5% (molecular weight 50-100kDa, sulfate content ≥25%)

[0084] Photoresponsive transdermal factor: 0.3% (4-dimethylaminocinnamic acid ethyl ester-cholesterol sulfate conjugate) Bionic sebum repair system: 10% (ceramide NP 2%, cholesterol 3%, fatty acid 5%)

[0085] Antioxidant Booster: 1% (Resveratrol Ferulate)

[0086] Slow-release preservative: 0.4% (caprylhydroxamic acid 0.24%, glyceryl caprylate 0.16%)

[0087] Balance: deionized water and pH adjuster (citric acid)

[0088] Preparation process:

[0089] (1) Active ingredient pretreatment:

[0090] Take 80 g of Epiphyllum extract and 400 g of β-cyclodextrin, add ethanol-water (1:4) mixed solvent, stir magnetically at 40°C for 5 hours, remove the solvent by rotary evaporation and freeze-dry to obtain a nano inclusion complex (inclusion rate 82%).

[0091] (2) Microsphere preparation:

[0092] Melatonin (10 g) and RGD tripeptide (5 g) were dissolved in 1000 mL of ethanol-water (1:1) solution, 2000 mL of 2% chitosan solution (pH 5.5) was added, and 500 mL of 1% sodium tripolyphosphate solution was added dropwise under 40 kHz ultrasonic conditions. After cross-linking for 30 minutes, the mixture was centrifuged (10,000 rpm, 15 min) and freeze-dried to obtain microspheres with a particle size of 110 nm.

[0093] (3) Oil phase preparation:

[0094] Dissolve 20 g of ceramide NP, 30 g of cholesterol, 50 g of fatty acid, and 3 g of light response factor in 500 mL of caprylic / capric triglyceride and stir at 70°C until completely dissolved.

[0095] (4) Preparation of aqueous phase:

[0096] Dissolve 20 g of nicotinamide mononucleotide and 10 g of pterostilbene in 3000 mL of deionized water, add 5 g of fucoidan, 10 g of antioxidant enhancer and preservative, and stir at 50°C until clear.

[0097] (5) Emulsification process:

[0098] The oil phase was slowly poured into the water phase, and homogenized and emulsified at 8000 rpm for 10 minutes to form a milky white emulsion with a particle size of 250-350 nm.

[0099] (6) Microsphere loading:

[0100] The temperature was lowered to 38°C, 200 mL of microsphere suspension and Epiphyllum nanoinclusion complex were added, ultrasonic dispersion was performed at 30 kHz for 15 minutes, and then stirred for 30 minutes.

[0101] (7) Finished product preparation:

[0102] Adjust the pH to 6.2 with citric acid, filter through a 0.45 μm filter membrane, and aseptically fill in brown glass bottles.

[0103] Performance test results:

[0104] Circadian rhythm regulation: BMAL1 gene expression increased by 40% (qRT-PCR detection)

[0105] Improved cellular metabolism: NAD+ Content increased by 50% (HPLC)

[0106] Barrier function: Transepidermal water loss reduced by 30% (measured by TEWL instrument)

[0107] Transdermal absorption: Quercetin transdermal efficiency reaches 42% (Franz diffusion cell method)

[0108] Safety: 0 allergic reactions in 30 patch tests

[0109] Example 3

[0110] Raw material formula (weight percentage):

[0111] Epiphyllum stem cell rhythm regulation extract: 15% (containing quercetin 16%, gamma-aminobutyric acid 0.3%)

[0112] Melatonin targeted delivery microspheres: 3% (chitosan embedded, RGD peptide density 9×10 9 molecules / μm2)

[0113] NAD + Precursor complex: 5% (nicotinamide mononucleotide: pterostilbene = 2:1)

[0114] Fucoidan: 2% (molecular weight 50-100kDa, sulfate content ≥25%)

[0115] Photoresponsive transdermal factor: 1% (4-dimethylaminocinnamic acid ethyl ester-cholesterol sulfate conjugate) Bionic sebum repair system: 18% (ceramide NP 4%, cholesterol 5%, fatty acid 8%)

[0116] Antioxidant Booster: 3% (Resveratrol Ferulate)

[0117] Slow-release preservative: 0.5% (caprylhydroxamic acid 0.3%, glyceryl caprylate 0.2%)

[0118] Balance: deionized water and pH adjuster (citric acid)

[0119] Preparation process:

[0120] (1) Active ingredient pretreatment:

[0121] Take 150g of Epiphyllum extract and 750g of β-cyclodextrin, add ethanol-water (1:4) mixed solvent, stir magnetically at 55°C for 3 hours, remove the solvent by rotary evaporation and freeze-dry to obtain a nano inclusion complex (inclusion rate 90%).

[0122] (2) Microsphere preparation:

[0123] Melatonin (30 g) and RGD tripeptide (15 g) were dissolved in 1000 mL of ethanol-water (1:1) solution, 2000 mL of 2% chitosan solution (pH 5.5) was added, and 500 mL of 1% sodium tripolyphosphate solution was added dropwise under 40 kHz ultrasonic conditions (power 250 W). After cross-linking for 30 minutes, the mixture was centrifuged (10,000 rpm, 15 min) and freeze-dried to obtain microspheres with a particle size of 85 nm.

[0124] (3) Oil phase preparation:

[0125] 40 g of ceramide NP, 50 g of cholesterol, 80 g of fatty acid and 10 g of light response factor were dissolved in 500 mL of caprylic / capric triglyceride and stirred at 75° C. until completely dissolved.

[0126] (4) Preparation of aqueous phase:

[0127] Dissolve 50 g of nicotinamide mononucleotide and 25 g of pterostilbene in 3000 mL of deionized water, add 20 g of fucoidan, 30 g of antioxidant synergist and preservative, and stir at 55°C until clear.

[0128] (5) Emulsification process:

[0129] The oil phase was slowly poured into the water phase, and homogenized and emulsified at 10,000 rpm for 10 minutes, followed by high-pressure homogenization (100 MPa, 2 times) to finally form a milky white emulsion with a particle size of 150-200 nm.

[0130] (6) Microsphere loading:

[0131] The temperature was lowered to 38°C, 200 mL of microsphere suspension and Epiphyllum nanoinclusion complex were added, ultrasonic dispersion was performed at 30 kHz for 15 minutes, and then stirred for 30 minutes.

[0132] (7) Finished product preparation:

[0133] Adjust the pH to 5.5 with citric acid, filter through a 0.45μm filter membrane, and aseptically fill in brown glass bottles. Performance test results:

[0134] Circadian rhythm regulation: BMAL1 gene expression increased by 60% (qRT-PCR detection)

[0135] Improved cellular metabolism: NAD + Content increased by 80% (HPLC)

[0136] Barrier function: Transepidermal water loss reduced by 45% (measured by TEWL instrument)

[0137] Transdermal absorption: Quercetin transdermal efficiency reaches 62% (Franz diffusion cell method)

[0138] Safety: 0 allergic reactions in 30 patch tests

[0139] Comparative Example

[0140] Raw material formula (weight percentage):

[0141] Epiphyllum japonica extract: 12% (containing quercetin 14.2%)

[0142] Unencapsulated melatonin: 0.5%

[0143] Niacinamide: 3.5%

[0144] Common red algae polysaccharide: 1.2%

[0145] Chemical penetration enhancer (propylene glycol): 2%

[0146] Traditional humectant (glycerin): 10%

[0147] Preservative (phenoxyethanol): 1%

[0148] Balance: deionized water and pH adjuster (citric acid)

[0149] Preparation process:

[0150] (1) Active ingredient pretreatment:

[0151] The epiphyllum extract was directly dissolved in the aqueous phase without inclusion treatment.

[0152] (2) Raw material mixing:

[0153] Melatonin and niacinamide were added directly to the aqueous phase without the use of microsphere or complex technology.

[0154] (3) Oil phase preparation:

[0155] Only glycerin dissolved in deionized water, no biomimetic sebum ingredients.

[0156] (4) Emulsification process:

[0157] The mixture was simply stirred without homogenization or sonication.

[0158] (5) Finished product preparation:

[0159] Adjust the pH to 5.8 with citric acid, filter through a 0.45 μm filter membrane and then fill.

[0160] Performance test results:

[0161] Circadian rhythm regulation: BMAL1 gene expression increased by 18% (qRT-PCR detection)

[0162] Improved cellular metabolism: NAD + Content increased by 22% (HPLC)

[0163] Barrier function: Transepidermal water loss reduced by 12% (measured by TEWL instrument)

[0164] Transdermal absorption: Quercetin transdermal efficiency is only 15% (Franz diffusion cell method)

[0165] Safety: 2 cases of mild erythema were found in 30 patch tests

[0166] The comparison of core gene expression and NAD+ levels between the examples and the comparative examples is shown in the following table:

[0167] Table 1

[0168] sample Example 1 Example 2 Example 3 Comparative Example BMAL1 mRNA elevation rate 52％ 40％ 60％ 18％ PER2 mRNA reduction rate 45％ 32％ 50％ 15％ NAD+ content increase rate 68％ 50％ 80％ 22％ COLLAGENⅠmRNA enhancement rate 55％ 42％ 68％ 18％

[0169] Conclusion: The embodiment significantly upregulated the expression of BMAL1 and COLLAGENⅠ genes, downregulated the PER2 gene, and increased the NAD+ level through the synergistic effect of Epiphyllum extract and targeted microspheres. The control group lacked rhythmic regulatory components, and the gene regulation efficiency was less than 40% of that of the embodiment.

[0170] The comparison of skin barrier repair and transdermal efficiency between the examples and the comparative examples is shown in the following table:

[0171] Table 2

[0172] sample Example 1 Example 2 Example 3 Comparative Example Transepidermal water loss (TEWL) reduction rate 38％ 30％ 45％ 12％ Transdermal efficiency (quercetin) 55％ 42％ 62％ 15％ Biomimetic lipid membrane thickness (nm) 65 58 78 No obvious membrane structure Irritation (patch positive rate) 0％ 0％ 0％ 10％

[0173] Conclusion: The biomimetic sebum repair system of the embodiment significantly reduces TEWL, and the light-responsive transdermal factor increases the absorption efficiency of the ingredients by 3-4 times without irritation. The comparative example relies on traditional penetration enhancers, has weak barrier repair ability and poses an irritation risk.

[0174] While embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions, and variations may be made to these embodiments without departing from the principles and spirit of the invention, and that the scope of the invention is defined by the appended claims and their equivalents.

Claims

1. A composition of Epiphyllum cyrtonema extract based on circadian rhythm gene regulation, characterized in that: By weight percentage, the raw materials include: Epiphyllum stem cell rhythm regulation extract: produced through supercritical CO2 extraction combined with β-cyclodextrin inclusion technology, containing quercetin, isorhamnetin and circadian rhythm regulating peptide precursors, which form a host-guest complex after β-cyclodextrin inclusion; Melatonin targeted delivery microspheres: Melatonin and RGD tripeptide are encapsulated by chitosan, forming a core-shell structure through electrostatic adsorption. The RGD tripeptide is arginine-glycine-aspartic acid. NAD + Precursor complex: Nicotinamide mononucleotide and pterostilbene are bound by hydrogen bonds, with a molecular weight of 635.6Da; Telomere-protecting polysaccharide: Fucoidan extracted from deep-sea red algae, with a sulfate content ≥ 25%, specifically binds to the telomerase reverse transcriptase subunit; Photoresponsive transdermal factor: A conjugate of ethyl 4-dimethylaminocinnamate and cholesterol sulfate, which can form transient holes under 365nm ultraviolet light; the photoresponsive transdermal factor coupling reaction formula is: Bionic sebum repair system: contains ceramide NP, cholesterol, and fatty acids in a molar ratio of 1:1:1; Antioxidant synergist: Resveratrol and ferulic acid are connected by ester bonds to form a bisphenol antioxidant molecule; Sustained-release preservative: a compound of caprylhydroxamic acid and glyceryl caprylate, which is slowly released through liposome encapsulation; The balance is deionized water and a pH regulator, and the pH regulator is citric acid or triethanolamine.

2. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The Epiphyllum stem cell rhythm regulation extract also contains gamma-aminobutyric acid (GABA), which can enhance the expression of the circadian rhythm gene BMAL1 by activating GABAA receptors.

3. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The melatonin targeted delivery microspheres are prepared by the following method: dissolving melatonin and RGD tripeptide in ethanol-water, adding chitosan solution, adding sodium tripolyphosphate solution dropwise under ultrasonic conditions, cross-linking to form microspheres, collecting by centrifugation and then freeze-drying.

4. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The NAD + In the precursor complex, the molar ratio of nicotinamide mononucleotide to pterostilbene is 2:1, and it is prepared into an amorphous powder by freeze-drying, and the particle size after reconstitution is ≤200nm.

5. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The added amount of the light-responsive transdermal factor is 0.5%.

6. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The purity of ceramide NP in the bionic sebum repair system is ≥98%, and the fatty acid is a mixture of oleic acid and linoleic acid in a ratio of 2:1, which can form a bionic lipid film with a thickness of 50-80 nm on the skin surface.

7. The epiphyllum extract composition based on circadian rhythm gene regulation according to claim 1, characterized in that The antioxidant activity of the antioxidant synergist is greater than or equal to 90%, and the activity retention rate after storage at 45° C. for 3 months is greater than or equal to 85%.

8. A method for preparing the Epiphyllum cyrtonema extract composition based on circadian rhythm gene regulation according to any one of claims 1 to 7, characterized in that: The following steps are involved: S1: Active ingredient pretreatment: Epiphyllum stem cell rhythm regulation extract and β-cyclodextrin were mixed in a ratio of 1:5 and stirred at 50°C for 4 hours to form a nanoinclusion complex; S2: Microsphere preparation: Prepare melatonin targeted delivery microspheres and disperse them in deionized water to form a stable suspension; S3: Oil phase preparation: Dissolve the biomimetic sebum repair system components and light-responsive transdermal factors in caprylic / capric triglyceride and stir evenly; S4: Aqueous phase preparation: NAD + The precursor complex, telomere-protecting polysaccharide, and antioxidant synergist are dissolved in deionized water, and a slow-release preservative is added; S5: Emulsification: Slowly add the oil phase to the water phase and emulsify in a homogenizer for 10 minutes to form an oil-in-water emulsion; S6: Microsphere loading: Cool to 35-40°C, add melatonin targeted delivery microsphere suspension and Epiphyllum nanoinclusion complex, and stir for 30 minutes; S7: pH adjustment and filling: Adjust the pH to 5.8-6.2 with citric acid, filter through a 0.45μm filter membrane, and aseptically fill in a light-proof container.

9. The method for preparing the epiphyllum extract composition based on circadian rhythm gene regulation according to claim 8, characterized in that: In step S6, 0.1-0.3% of gamma-aminobutyric acid was added and ultrasound-assisted dispersion was used for 15 minutes.

10. The method for preparing the Epiphyllum cyrtonema extract composition based on circadian rhythm gene regulation according to claim 8, characterized in that: The prepared composition needs to be irradiated with 365nm ultraviolet light for 3-5 minutes before use to activate the light-responsive transdermal factor and form a transient permeation channel.

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