PH response type anti-decline microsphere composition based on regulation and control of circadian rhythm and application of pH response type anti-decline microsphere composition

Abstract

The present invention relates to a circadian rhythm regulation-based pH responsive anti-decline microsphere composition and application thereof, the circadian rhythm regulation-based pH responsive anti-decline microsphere composition comprises an inner sodium alginate microsphere and an outer chitosan coating layer, the inner sodium alginate microsphere encapsulates berberine and hyaluronic acid; the outer chitosan coating layer is used for coating nicotinamide, resveratrol and the inner sodium alginate microspheres. The pH response type anti-decline microsphere composition is applied to a skin care product. Compared with the prior art, the preparation method has the advantages that the circadian rhythm adaptive release of active ingredients is realized by constructing a pH response type double-layer microsphere structure of'sodium alginate on the inner layer and chitosan on the outer layer '.

Description

Technical Field

[0001] This invention belongs to the field of cosmetics and functional skin care products, specifically relating to a pH-responsive anti-aging microsphere composition based on regulating diurnal rhythms and its application. Background Technology

[0002] As the largest organ in the human body, the skin's physiological activities are regulated by an endogenous biological clock, forming a circadian rhythm system with a 24-hour cycle. This system is driven by a transcription-translation feedback loop (TTFL) composed of core clock genes CLOCK, BMAL1, PER1 / 2, and CRY1 / 2, which achieves a dynamic diurnal balance of skin function by regulating the expression of downstream target genes. The CLOCK / BMAL1 heterodimer activates the transcription of Period (Per) and Cryptochrome (Cry) genes, and the PER / CRY protein complex, after entering the nucleus, feedback inhibits CLOCK / BMAL1 activity, forming an autonomously oscillating molecular rhythm. This rhythm regulation not only affects the energy metabolism of skin cells but is also directly related to key anti-aging physiological processes such as collagen synthesis, barrier repair, and inflammatory responses. Studies have confirmed that during the day, the skin mainly focuses on collagen fiber assembly, enhancing its resistance to mechanical stress and ultraviolet radiation; at night, it shifts to procollagen synthesis and DNA damage repair, with increased cell proliferation rate and doubled repair efficiency, exhibiting the physiological characteristics of "daytime defense and nighttime repair." It is worth noting that the pH value of the skin surface exhibits a physiological fluctuation from 5.5 during the day to 4.5 at night. This endogenous circadian rhythm can further drive the diurnal variation of barrier permeability and enzyme activity, providing an important target for circadian regulation.

[0003] The metabolic-circulatory clock bidirectional regulatory network plays a key role in the skin aging process, among which NAD+... + The diurnal oscillation of skin rhythms regulates CLOCK / BMAL1 activity by activating SIRT1 deacetylase. As a cellular energy metabolism sensor, AMPK (AMP-activated protein kinase) plays a pivotal role in this network—it phosphorylates downstream target proteins to synergistically regulate three major anti-aging pathways: mitochondrial biosynthesis (PGC-1α pathway), autophagy (ULK1 phosphorylation), and inflammation suppression (NF-κB inactivation). When skin rhythms are disrupted (e.g., due to lack of sleep or blue light exposure), decreased AMPK activity leads to mitochondrial dysfunction, reduced autophagy clearance efficiency, and excessive release of inflammatory factors, ultimately causing disordered collagen fiber accumulation and damage to the skin barrier, manifesting as loss of skin elasticity and wrinkle formation.

[0004] Although existing research confirms the anti-aging potential of AMPK activators, current technology faces significant bottlenecks: traditional anti-aging ingredients generally lack circadian rhythm adaptability, and their mode of action is mostly static release, which cannot adjust the active output according to the skin's dynamic needs of "daytime protection-nighttime repair", resulting in a mismatch between efficacy and physiological needs.

[0005] The diurnal fluctuations in skin surface pH serve as an external manifestation of the endogenous biological clock, constituting a potential circadian rhythm regulation signal. pH changes not only directly affect lipid arrangement and enzyme activity in the stratum corneum, but also act as a spatiotemporally specific "biological switch"—a pH gradient-responsive delivery system preferentially releases antioxidants in the near-neutral daytime environment and enhances the penetration of repairing ingredients in the acidic nighttime environment. However, most products still employ a single release mode, failing to effectively utilize this signal to build an intelligent response system, resulting in a bottleneck in anti-aging efficacy.

[0006] Patent CN120081881A discloses a glutamate derivative that achieves circadian rhythm regulation and antioxidant effects by upregulating CLOCK / Per3 protein expression, inhibiting ROS, and activating the Nrf2 pathway. However, its preparation process is highly complex, requiring amidation, deprotection, and multi-step extraction and purification, with stringent reaction conditions, increasing the cost of large-scale production. Furthermore, it lacks a circadian rhythm-adaptive delivery system; although it can regulate circadian rhythm genes, it does not utilize the diurnal pH fluctuations of the skin to design a carrier, resulting in a static release of the active ingredient. This fails to meet the dynamic needs of "daytime anti-oxidation - nighttime repair," potentially leading to diminished efficacy.

[0007] Patent CN119185124A discloses a composition containing circadian rhythm regulators, ascorbate glucoside, and other ingredients, which achieves whitening through anti-oxidation, anti-glycation, anti-inflammation, and circadian rhythm regulation. However, it relies on traditional formulations (aqueous solutions, emulsions, etc.) and does not construct a delivery system that responds to the skin's physiological rhythm. It cannot dynamically adjust the release of ingredients according to daytime / nighttime needs (e.g., preferentially releasing antioxidant ingredients during the day and enhancing the penetration of repair ingredients at night), resulting in insufficient activity utilization. Summary of the Invention

[0008] The purpose of this invention is to provide a pH-responsive anti-aging microsphere composition based on the regulation of diurnal rhythm and its application. By constructing a pH-responsive bilayer microsphere structure of "inner layer sodium alginate-outer layer chitosan", the diurnal rhythm adaptation release of active ingredients is achieved.

[0009] The objective of this invention can be achieved through the following technical solution: a pH-responsive anti-aging microsphere composition based on regulating diurnal rhythm, comprising an inner layer of sodium alginate microspheres and an outer layer of chitosan coating, wherein the inner layer of sodium alginate microspheres encapsulates berberine and hyaluronic acid; the outer layer of chitosan coating encapsulates nicotinamide, resveratrol, and the inner layer of sodium alginate microspheres.

[0010] Furthermore, the mass ratio of sodium alginate, berberine, and hyaluronic acid is (1~3):(0.5~1.5):(0.5~1.5). The mass ratio of the inner layer sodium alginate microspheres, nicotinamide, and resveratrol is (5~15):(0.5-1.5):(1.5-2.5).

[0011] Furthermore, the inner layer sodium alginate microspheres are prepared by the following method: S11. Add sodium alginate to deionized water and stir until completely dissolved to form a clear solution; S12. Add berberine and hyaluronic acid to the solution obtained in step S11, stir and mix evenly to obtain an aqueous phase; S13. The aqueous phase obtained in step S12 is slowly dripped into the oil phase containing the emulsifier, and a uniform W / O emulsion is formed by high-speed shear emulsification. S14. Add calcium chloride solution dropwise to the emulsion obtained in step S13 and stir to promote cross-linking of sodium alginate molecules into spheres. S15. Centrifuge the product obtained in step S14, collect the precipitated microspheres, wash with deionized water and freeze-dry to obtain inner layer microsphere powder loaded with berberine and hyaluronic acid.

[0012] Further, in step S11, the mass-to-volume ratio of sodium alginate to deionized water is 1~3:100 (w / v), and the stirring conditions are 45~55℃ and 250~350rpm, until completely dissolved to form a clear solution.

[0013] In step S12, the mass concentrations of berberine and hyaluronic acid are 0.5~1.5% w / v and 0.5~1.5% w / v, respectively, and the stirring conditions are: temperature 45-55℃, speed 450~550 rpm, and time 20~40 min.

[0014] In step S13, the oil phase is liquid paraffin containing 3-6% v / v Span 80, the volume ratio of the water phase to the oil phase is 1:4-6, and the high-speed shear emulsification conditions are a rotation speed of 1200-1600 rpm, a temperature of 35-45℃, and a time of 30-50 min. In step S14, the mass-volume concentration of the calcium chloride solution is 8-12%, and the volume ratio of the calcium chloride solution to the emulsion is 1:4-6. After the addition, the stirring conditions are: temperature 20-30℃, speed 150-250rpm, and time 50-70min. In step S15, the centrifugation conditions are 2500~3500 rpm for 8~12 min; the freeze-drying conditions are -45~-55℃ for 22~26 h.

[0015] Furthermore, the outer chitosan coating layer is prepared by the following method: S21. Dissolve chitosan in an acidic aqueous solution, add nicotinamide and resveratrol, and stir until completely dissolved to obtain a chitosan solution. S22. Add the inner layer sodium alginate microspheres to the chitosan solution obtained in step S21 and disperse them by ultrasonication to form a uniform aqueous phase; S23. Add the aqueous phase obtained in step S22 to the oil phase containing the emulsifier, and stir to emulsify and form a W / O secondary emulsion; S24. Add sodium tripolyphosphate solution dropwise to the secondary emulsion obtained in step S23 and stir to promote cross-linking and solidification of chitosan molecules; S25. Centrifuge the product obtained in step S24, collect the composite microspheres, wash them with anhydrous ethanol and deionized water, and freeze-dry them to obtain pH-responsive sustained-release microsphere solid powder.

[0016] Further, in step S21, the acidic aqueous solution is an acetic acid aqueous solution with a volume concentration of 0.5-1.5%, the chitosan dissolved in the acetic acid aqueous solution has a mass-volume concentration of 0.5-1.5% w / v, and the mass concentrations of nicotinamide and resveratrol are 0.5-1.5% w / v and 1.5-2.5% w / v, respectively. The stirring conditions are a temperature of 20-30℃, a rotation speed of 350-450 rpm, and a time of 10-30 min. In step S22, the mass-to-volume ratio of the inner sodium alginate microspheres to the chitosan solution is 5-15% w / v, and the ultrasonic dispersion conditions are 250-350 W power and 5-15 min time, carried out at room temperature. In step S23, the oil phase is liquid paraffin containing 2-4% v / v Tween 80, the volume ratio of the water phase to the oil phase is 1:3-5, and the stirring emulsification conditions are temperature 25-35℃, speed 1000-1500 rpm, and time 25-35 min. In step S24, the mass-volume concentration of sodium tripolyphosphate solution is 0.5-1.5% w / v, and the volume ratio of sodium tripolyphosphate solution to chitosan solution is 1:4-6. After the addition, the stirring conditions are: temperature 20-30℃, speed 550-650 rpm, and time 40-50 min. In step S25, the centrifugation conditions are 3500~4500 rpm for 10~20 min; the freeze-drying conditions are -45~-55℃ for 22~26 h.

[0017] The present invention also provides an application of a pH-responsive anti-aging microsphere composition based on regulating circadian rhythms, wherein the pH-responsive anti-aging microsphere composition is used as an active ingredient in skin care products.

[0018] Furthermore, the skincare products mentioned include skincare water, serum, lotion, freeze-dried powder, or face cream.

[0019] Furthermore, the skincare product is a face cream, which comprises phase A, phase B, phase C, phase D, and phase E, each comprising the following components in weight percentage: Phase A includes: Glycerin 5-10%, Butanediol 3-10%, Trehalose 0.5-2%, Sodium hyaluronate 0.01-1%, Tocopheryl acetate 0.01-1%, Panthenol 0.1-5%, Hydroxypropyl methylcellulose 0.1-2%, Sodium acrylate / sodium acryloyldimethyl taurate copolymer 0.1-2%, PEG-100 stearate 0.1-2%, EDTA disodium 0.01-1%, Water balance; Phase B includes: Isopropyl isostearate 0.1-5%, Caprylic / capric triglycerides 0.1-5%, Glyceryl stearate 0.1-5%, Polydimethylsiloxane 0.1-4%, Cetearyl alcohol 0.1-5%, behenol 0.1-2%, Sorbitan oleate 0.1-2%; Phase C includes: Arginine 0.1-2%; Phase D includes: Active ingredient 1-5%; Phase E includes: (Daily use) Fragrance 0.1-2%; Phenoxyethanol 0.01-1%.

[0020] Furthermore, the face cream is prepared by the following method: (1) Aqueous phase preparation: Place water in a reactor, fully wet and mix the other components of phase A, add them to the water, stir thoroughly until completely dissolved, and heat to 75-80℃; (2) Preparation of oil phase: Mix the components of phase B and heat to 75-80℃, stirring until completely dissolved; (3) Aqueous phase-oil phase mixing: Slowly pour the oil phase prepared in step (2) into the aqueous phase in step (1), stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min; (4) Add each component in phase C, stir at 250-350 rpm for 3-5 min, then homogenize at 2500-3500 rpm for 3-10 min, and stir to cool to 40-50℃; (5) Add each component in phase D, stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min; (6) After cooling to 35-38℃, add phase E, stir at 250-350 rpm for 3-10 min, then homogenize at 2500-3500 rpm for 3-10 min to obtain face cream.

[0021] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention achieves diurnal rhythm-adaptive release of active ingredients by constructing a pH-responsive bilayer microsphere structure of "inner layer sodium alginate - outer layer chitosan". The inner sodium alginate microspheres encapsulate berberine and hyaluronic acid, while the outer chitosan coating layer loads nicotinamide and resveratrol. This microsphere carrier utilizes the diurnal pH fluctuations of the skin (neutral 5.5-7.0 during the day / acidic 4.5-5.0 at night) to achieve dynamic release regulation. Daytime protection phase: Under neutral conditions, the outer chitosan gradually dissolves, and the nicotinamide and resveratrol loaded on it are slowly released, activating the Nrf2 pathway to scavenge free radicals; at the same time, the inner sodium alginate microspheres swell moderately under neutral conditions, synergistically inhibiting NF-κB inflammatory signals, achieving free radical scavenging and UV damage protection.

[0022] Nighttime repair phase: The acidic ring accelerates the swelling of the chitosan layer, speeds up the release rate of nicotinamide and resveratrol, and exposes the inner sodium alginate microspheres, releasing berberine and hyaluronic acid to synergistically repair daytime damage and enhance the stratum corneum's water-locking ability.

[0023] (2) The rhythmic release mode of "daytime protection-nighttime repair" of this invention effectively avoids the activity decay of free components due to pH fluctuations, and solves the technical bottleneck that traditional anti-aging products cannot adapt to the physiological rhythm of the skin. Attached Figure Description

[0024] Figure 1 Facial redness map of subjects at different times before using the face cream described in Example 1. Detailed Implementation

[0025] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0026] This invention constructs a pH-responsive bilayer microsphere structure consisting of an inner layer of sodium alginate and an outer layer of chitosan, comprising resveratrol, berberine, hyaluronic acid, and niacinamide. The resveratrol, berberine, hyaluronic acid, and niacinamide are prepared into pH-responsive sustained-release microspheres. This allows for adjustment of active output based on the dynamic needs of the skin during "daytime protection and nighttime repair," thus adapting the efficacy to physiological requirements.

[0027] The outer chitosan coating layer is loaded with niacinamide and resveratrol. Niacinamide is the amide form of vitamin B3 and exhibits excellent stability in neutral to slightly acidic environments. Niacinamide can inhibit the expression and activation of PAR-2 ​​receptors (protease-activated receptor-2) on the surface of keratinocytes, reduce the transfer of melanosomes from melanocytes to keratinocytes, and activate the AMPK signaling pathway, promoting keratinocyte proliferation and shedding, shortening the epidermal renewal cycle, accelerating the removal of melanin-containing keratinocytes, and improving uneven skin tone and dullness. Furthermore, niacinamide can activate the Sirtuins protein family, upregulate the mRNA expression of type I and type III collagen and elastin, reduce the degradation of collagen by matrix metalloproteinases in the dermis, and improve skin elasticity. Resveratrol is a naturally occurring non-flavonoid polyphenol compound, primarily derived from the roots, fruits, or seeds of plants in the Vitaceae, Polygonaceae, and Leguminosae families. Its phenolic hydroxyl structure can directly provide hydrogen atoms to neutralize free radicals, inhibiting lipid peroxidation to reduce oxidative damage to cell membranes and DNA. Simultaneously, it enhances the cellular antioxidant defense system by upregulating the Nrf2 transcription factor, promoting the expression of endogenous antioxidant enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). In terms of anti-aging, resveratrol promotes DNA repair and prolongs cell lifespan by activating the SIRT1 pathway, while inhibiting the degradation of collagen by matrix metalloproteinases (MMP-1, MMP-3) and activating the AMPK energy metabolism pathway to improve mitochondrial function, reducing skin laxity and wrinkle formation. Furthermore, it can alleviate inflammatory responses such as erythema and edema caused by ultraviolet radiation, environmental pollution, or mechanical stimulation by inhibiting the NF-κB inflammatory signaling pathway and reducing the release of pro-inflammatory factors such as TNF-α and IL-6, thus reducing sensitivity and discomfort.

[0028] When the skin surface is neutral during the day (pH 5.5-7.0), the outer chitosan slowly dissolves, gradually releasing nicotinamide and resveratrol. Through the synergistic effect of Nrf2 pathway activation and NF-κB inflammation inhibition, it achieves antioxidant and daytime protection. The inner layer of sodium alginate microspheres encapsulates berberine and hyaluronic acid: Berberine is a natural isoquinoline alkaloid, mainly derived from the rhizomes or bark of plants in the Ranunculaceae family such as Coptis chinensis, Phellodendron chinense, and Berberis vulgaris. In terms of anti-inflammatory and soothing effects, it inhibits IκB kinase activity in the NF-κB signaling pathway, preventing p65 subunit nucleation to reduce the release of pro-inflammatory factors such as IL-1β and TNF-α, and downregulates the phosphorylation levels of p38 and JNK in the MAPK pathway, reducing the production of inflammatory mediators such as prostaglandin E2 and lowering the skin sensitivity threshold. Regarding its antioxidant defense mechanism, berberine can directly scavenge superoxide anions and hydroxyl radicals through its phenolic hydroxyl groups, inhibiting the production of lipid peroxidation product MDA, while simultaneously activating the Nrf2 / HO-1 pathway, promoting the expression of antioxidant molecules such as heme oxygenase-1 and glutathione, and enhancing the cell's resistance to exogenous oxidative stimuli. In terms of repairing the skin barrier and enhancing skin homeostasis, it can activate the AMPK / mTOR signaling pathway, accelerate the proliferation and differentiation of keratinocytes, increase the synthesis of filaggrin and natilbene to strengthen the physical barrier, and regulate the skin microecological balance by selectively inhibiting harmful bacteria and promoting the colonization of beneficial bacteria, thereby reducing the risk of inflammation recurrence.

[0029] Hyaluronic acid is a linear glycosaminoglycan composed of D-glucuronic acid and N-acetylglucosamine linked by alternating glycosidic bonds, primarily prepared through microbial fermentation. Hyaluronic acid can penetrate the granular and spinous layers, promoting hydration and swelling of keratinocytes through osmotic pressure regulation, improving dryness symptoms such as rough skin and flaking, and upregulating aquaporin 3 (AQP3) expression, enhancing the efficiency of intracellular and extracellular water transport. It binds to the CD44 receptor on the surface of keratinocytes, activating the PI3K / Akt signaling pathway to promote cell proliferation and migration. Simultaneously, it upregulates the expression of filaggrin and natilbene to increase the content of natural moisturizing factors, and stimulates ceramide synthase activity by activating the PPAR-γ pathway, improving the integrity of the stratum corneum lipid bilayer and effectively repairing the skin barrier. Furthermore, hyaluronic acid competitively binds to the TLR4 receptor, blocking NF-κB pathway activation, reducing the release of pro-inflammatory factors such as IL-1β and TNF-α, and inhibiting mast cell degranulation to lower the skin sensitivity threshold.

[0030] At night, when the skin becomes acidic (pH 4.5-5.0), the chitosan layer swells and accelerates the release of niacinamide and resveratrol, exposing the inner sodium alginate microspheres, which release berberine and hyaluronic acid. These, along with niacinamide and resveratrol, synergistically enhance the stratum corneum's water-locking capacity and barrier repair efficiency. This rhythmic release pattern of "daytime protection - nighttime repair" effectively avoids the activity decay of free components due to pH fluctuations, overcoming the technical bottleneck of traditional anti-aging products' inability to adapt to the skin's physiological rhythm.

[0031] Furthermore, the aforementioned active ingredients of this invention can be used in various skincare products, including but not limited to toners, serums, lotions, creams, and freeze-dried powders. The required base ingredients vary depending on the type of skincare product. The base ingredients can be selected from conventional base ingredients used in existing products, and the skincare product can be prepared using conventional methods. The resulting skincare product has a gentle anti-aging effect.

[0032] Taking face cream as an example, the pH-responsive anti-aging microsphere composition based on regulating circadian rhythm of this invention is formulated into a face cream together with the base ingredients. The specific formula is shown in Table 1 below: Table 1. Proportions of each component in the face cream formula.

[0033] Face cream is made using the following methods: Preparation of S1 inner layer microspheres: S11. Aqueous phase preparation: Dissolve sodium alginate in deionized water and stir continuously at 45~55℃ and 250~350rpm until completely dissolved. S12. Then add berberine and hyaluronic acid, and stir magnetically for 20-40 minutes at 45-55℃ and 450-550 rpm to form a homogeneous aqueous solution.

[0034] The amount of each raw material added was such that the concentrations of each raw material in the resulting aqueous solution were 1-3% w / v sodium alginate, 0.5-1.5% w / v berberine, and 0.5-1.5% w / v hyaluronic acid.

[0035] S13. Emulsification reaction: The prepared aqueous phase is slowly dripped into liquid paraffin containing Span 80 (the volume ratio of aqueous phase to oil phase is 1:4~6, preferably 1:5). Emulsification is carried out at 1200~1600 rpm, preferably 1500 rpm, 35~45℃ for 30~50 min, preferably 40 min, to form a stable W / O emulsion. The oil phase is liquid paraffin, the emulsifier is 3~6% v / v Span 80, and the crosslinking agent is 8~12% w / v calcium chloride solution. S14, Ionic Crosslinking: Add calcium chloride solution dropwise to the emulsion (volume ratio of calcium chloride solution to emulsion is 1:4~6), and stir continuously at 20~30℃ and 150~250 rpm for 50~70 min, preferably 1 hour, through Ca... 2+ The cross-linking with sodium alginate solidifies it, forming inner microspheres loaded with berberine and hyaluronic acid.

[0036] S15. Separation and washing: Centrifuge the solidified microspheres at 2500~3500 rpm, preferably 3000 rpm for 8~12 min, preferably 10 min. After collection, wash them three times each with petroleum ether, anhydrous ethanol and deionized water to remove residual oil phase and free calcium chloride. Finally, freeze-dry them under vacuum at -45~-55℃, preferably -50℃ for 22~26 h, preferably 24 h for later use.

[0037] The S2 outer layer is coated, and the specific steps are as follows: S21. Preparation of the outer aqueous phase: Chitosan is dissolved in an aqueous acetic acid solution (the volume concentration of the aqueous acetic acid solution is 0.5-1.5%) and stirred until clear and transparent. The mass volume concentration of chitosan dissolved in the aqueous acetic acid solution is 0.5-1.5% w / v. Then, nicotinamide and resveratrol ethanol solution are added, and the mixture is magnetically stirred at 20-30℃ and 350-450 rpm, preferably 400 rpm, for 10-30 min, preferably 20 min, to form a chitosan solution containing the active ingredient with a pH of 5.0-5.5. The mass concentrations of nicotinamide and resveratrol in this solution are 0.5-1.5% w / v and 1.5-2.5% w / v, respectively.

[0038] S22. Secondary emulsification: Disperse the inner layer sodium alginate microspheres in the chitosan solution obtained in step S21 above. The mass-to-volume ratio of the inner layer sodium alginate microspheres to the chitosan solution is 5-15% w / v. The solution is ultrasonically treated at 250-350W, preferably 300W, for 5-15 minutes, preferably 10 minutes, at room temperature to ensure uniform dispersion, which is used as the aqueous phase. S23. Add the aqueous phase to the liquid paraffin containing Tween 80 (i.e., the oil phase, which is liquid paraffin containing 2-4% v / v Tween 80, with a volume ratio of aqueous phase to oil phase of 1:3-5, preferably 1:4), and stir and emulsify for 25-35 min at 1000-1500 rpm, preferably 1200 rpm, and at a temperature of 25-35℃, preferably 30℃, to form a W / O secondary emulsion.

[0039] S24. Outer layer cross-linking and curing: Add sodium tripolyphosphate solution (TPP solution with a mass-volume concentration of 0.5-1.5% w / v and a volume ratio of 1:4 to 6, preferably 1:5) to the secondary emulsion. Stir at 20-30℃ and 550-650 rpm for 40-50 min, preferably 45 min. Through electrostatic interaction, chitosan and TPP are cross-linked to form an outer layer coating structure.

[0040] S25. Separation and purification: The composite microspheres obtained in step S23 are centrifuged at 3500~4500 rpm, preferably 4000 rpm for 10~20 min, preferably 15 min. After collection, they are washed twice with anhydrous ethanol and three times with deionized water to remove unencapsulated components and residual emulsifiers. Finally, they are freeze-dried at -45~-55℃, preferably -50℃, for 22~26 h, preferably 24 h, to obtain pH-responsive sustained-release microspheres.

[0041] S3 face cream is prepared using the following method: S31, Aqueous phase: Place water in a container equipped with a stirring device and stir. After fully wetting and mixing the other components of phase A, add them to the water and stir thoroughly until completely dissolved. Heat to 75-80℃. S32, oil phase; mix the components of phase B and heat to 75-80℃, stirring until completely dissolved; S33. Emulsification: Add the oil phase obtained in step S32 to the aqueous phase obtained in step S31, stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min. S34. Add each component in phase C, stir at 250-350 rpm for 3-5 min, then homogenize at 2500-3500 rpm for 3-10 min, and stir while cooling to 40-50℃. S35. Add each component in phase D, stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min. S36. After cooling to 35-38℃, add phase E, stir at 250-350 rpm for 3-10 minutes, then homogenize at 2500-3500 rpm for 3-10 minutes to obtain the desired face cream.

[0042] The following detailed description is provided through specific embodiments and comparative examples.

[0043] The raw material composition of Examples 1-3, Comparative Examples 1-7, and Blank Example 1 is shown in Table 2 below: Table 2. Raw material composition ratios of Examples 1-3, Comparative Examples 1-7, and Blank Example 1.

[0044] The preparation method of the above face cream is as follows: Preparation of S1 inner layer microspheres: S11. Aqueous phase preparation: Dissolve sodium alginate in deionized water and stir continuously at 50°C and 300 rpm until completely dissolved. S12. Berberine and hyaluronic acid were then added, and the mixture was magnetically stirred at 500 rpm for 30 minutes to form a homogeneous aqueous solution. The amounts of each raw material added were such that the concentrations of each raw material in the resulting aqueous solution were 2% w / v sodium alginate, 1% w / v berberine, and 1% w / v hyaluronic acid, respectively.

[0045] S13. Emulsification reaction: The prepared aqueous phase is slowly dripped into liquid paraffin containing Span 80 (the volume ratio of aqueous phase to oil phase is 1:5). Emulsification is carried out at 1500 rpm and 40℃ for 40 min to form a stable W / O emulsion. The oil phase is liquid paraffin, the emulsifier is 5% v / v Span 80, and the crosslinking agent is 10% w / v calcium chloride solution. S14, Ionic Crosslinking: Calcium chloride solution (volume ratio of calcium chloride solution to emulsion 1:5) is added dropwise to the emulsion, and the mixture is stirred continuously at 200 rpm for 60 min. The crosslinking is achieved through the Ca... 2+ The cross-linking with sodium alginate solidifies it, forming inner microspheres loaded with berberine and hyaluronic acid.

[0046] S15. Separation and washing: Centrifuge the solidified microspheres at 3000 rpm for 10 min, collect them and wash them three times each with petroleum ether, anhydrous ethanol and deionized water to remove residual oil phase and free calcium chloride. Finally, freeze dry them at -50℃ for 24 h for later use.

[0047] S2 outer layer coating The specific steps are as follows: S21. Preparation of the outer aqueous phase: Chitosan is dissolved in an aqueous acetic acid solution (the volume concentration of the aqueous acetic acid solution is 1%) and stirred until clear and transparent. The mass volume concentration of chitosan dissolved in the aqueous acetic acid solution is 1% w / v. Then, nicotinamide and resveratrol ethanol solution are added, and the mixture is magnetically stirred at 400 rpm for 20 min to form a chitosan solution with a pH of 5.0~5.5 containing the active ingredient. The mass concentrations of nicotinamide and resveratrol in this solution are 1% w / v and 2% w / v, respectively.

[0048] S22, Secondary emulsification: The inner layer sodium alginate microspheres are dispersed in the chitosan solution obtained in step S21 above. The mass-to-volume ratio of the inner layer sodium alginate microspheres to the chitosan solution is 5~15%w / v. The solution is ultrasonically treated at 300 W for 10 min to make it uniformly dispersed, and this solution is used as the aqueous phase. S23. Add the aqueous phase to the liquid paraffin containing Tween 80 (i.e., the oil phase, which is liquid paraffin containing 3% v / v Tween 80, with a volume ratio of 1:4 between the aqueous phase and the oil phase), and stir and emulsify at 1200 rpm and 30°C for 30 min to form a W / O secondary emulsion.

[0049] S24. Outer layer cross-linking and curing: Add sodium tripolyphosphate solution (TPP solution with a mass-volume concentration of 1% w / v and a volume ratio of 1:5 with chitosan solution) dropwise to the secondary emulsion, stir at 600 rpm for 45 min, and cross-link chitosan and TPP through electrostatic interaction to form an outer layer coating structure.

[0050] S25. Separation and purification: The composite microspheres obtained in step S23 were centrifuged at 4000 rpm for 15 min, collected, and washed twice with anhydrous ethanol and three times with deionized water to remove unencapsulated components and residual emulsifiers. Finally, they were freeze-dried at -50℃ for 24 h to obtain pH-responsive sustained-release microspheres.

[0051] S3 face cream is prepared using the following method: S31, Aqueous phase: Place water in a container equipped with a stirring device and stir. After fully wetting and mixing the other components of phase A, add them to the water and stir thoroughly until completely dissolved. Heat to 75-80℃. S32, oil phase; mix the components of phase B and heat to 75-80℃, stirring until completely dissolved; S33. Emulsification: Add the oil phase obtained in step S32 to the aqueous phase obtained in step S31, stir at 300 rpm for 4 min, and then homogenize at 3000 rpm for 6 min. S34. Add each component in phase C, stir at 300 rpm for 4 min, then homogenize at 3000 rpm for 6 min, and stir to cool to 45℃. S35. Add each component in phase D, stir at 300 rpm for 4 min, and then homogenize at 3000 rpm for 6 min. S36. After cooling to 35-38℃, add phase E, stir at 300rpm for 6 minutes, then homogenize at 3000rpm for 6 minutes to obtain the desired face cream.

[0052] The only difference between Example 2 and Example 1 is the addition of each active ingredient, and the only difference between Example 3 and Example 1 is the reduction of each active ingredient.

[0053] Comparative Example 1 is chitosan microspheres encapsulating niacinamide and resveratrol; that is, without step S1 above, without adding sodium alginate microspheres in step S22, and otherwise the same as step S2 to obtain chitosan microspheres encapsulating niacinamide and resveratrol, which are used as the active ingredient in phase D in face cream preparation S3. Comparative Example 2 is sodium alginate microspheres encapsulating berberine and hyaluronic acid; that is, without step S2 above, the sodium alginate microspheres obtained in step S1 are directly used as the active ingredient of phase D in the preparation of face cream S3. Comparative Examples 3-6 contain only one of the four free components: nicotinamide, resveratrol, berberine, and hyaluronic acid; that is, steps S1 and S2 are not included. When adding phase D components in step S3, one of the four components, namely nicotinamide, resveratrol, berberine, and hyaluronic acid, is added as phase D components in step S35. Comparative Example 7 contains four free components: nicotinamide, resveratrol, berberine, and hyaluronic acid. That is, it does not include steps S1 and S2. When adding phase D components in step S3, nicotinamide, resveratrol, berberine, and hyaluronic acid are added as phase D components in step S35.

[0054] Blank Example 1 is a face cream that does not contain any active ingredients, i.e., it does not contain phase D.

[0055] Performance testing: I. Cell experiments were conducted using the active ingredients from Examples 1-3, Comparative Examples 1-7, and Blank Example 1 as experimental samples, as follows: The proportions of cell experimental raw materials for each experimental example and control group are shown in Table 3 below: Table 3. Distribution ratio of raw materials for cell experiments

[0056] 1. ROS cleanup capability test (1) Experimental materials Reagents: DCFH-DA fluorescent probe (10 mM), H2O2 (100 μM, oxidative stress inducer), PBS buffer (pH 7.4), and sample solutions for each group (Examples 1-3 and Comparative Examples 1-7 in Table 3). Instrument: Fluorescence spectrophotometer (excitation wavelength 488 nm, emission wavelength 525 nm) (2) Experimental Procedure Sample pretreatment: Dilute each group of samples with PBS to a concentration of 0.1 mg / mL (Experimental Examples 1-3 and Comparative Examples 1-7) for later use; Probe loading: Add 100 μL of DCFH-DA probe (final concentration 10 μM) to a 96-well plate and incubate at 37°C in the dark for 30 min; Oxidative stress induction: Add H2O2 solution (final concentration 100 μM) and incubate for 2 h; Sample intervention: Add 100 μL of sample solutions of different concentrations and incubate at 37℃ for 1 h; Fluorescence detection: The fluorescence intensity of each group was measured using a fluorescence spectrophotometer, with 3 replicates per group.

[0057] Data Analysis H2O2 model group: The group with only H2O2 added and no sample added (blank control group), whose fluorescence intensity represents 100% ROS level; Experimental group: The group in which H2O2 and the sample were added; ROS scavenging rate was calculated as follows: the fluorescence intensity of the blank control group after treatment with the above experimental steps was used as the fluorescence intensity of the H2O2 model group; the other groups of samples were used as experimental groups. Scavenging rate (%) = [(fluorescence intensity of H2O2 model group − fluorescence intensity of experimental group) / (fluorescence intensity of H2O2 model group)] * 100%.

[0058] 2. DPPH free radical scavenging ability test (1) Experimental materials Reagents: DPPH ethanol solution (0.1 mM), anhydrous ethanol, Instrument: UV-Vis spectrophotometer (517 nm wavelength) Samples: The active ingredients shown in Table 4 below were used as samples. Table 4 Experimental ratio for DPPH free radical scavenging ability test

[0059] (2) Experimental Procedure Sample dilution: Prepare each group of samples with anhydrous ethanol to a concentration of 0.1 mg / mL (Examples 1-3 and Comparative Examples 1-7) for later use; Reaction system: Add 100 μL of DPPH solution to a 96-well plate, then add 100 μL of each group of sample solutions, mix well and let stand in the dark for 30 min. Blank control: Use anhydrous ethanol instead of the sample solution and place under the same conditions for 30 min; Absorbance measurement: The absorbance (A) of each group was measured at 517 nm.

[0060] (3) Data Analysis Calculate DPPH clearance rate: Clearance rate (%) = [(A) 空白 -A 样品 ) / A 空白 ]*100%.

[0061] Table 5. Results of ROS scavenging and DPPH free radical scavenging abilities

[0062] As shown in Table 5, the ROS / DPPH scavenging rates of Experiments 1-3 increased significantly with increasing concentration: the ROS scavenging rate of Experiment 2 reached 85.7% and the DPPH scavenging rate reached 82.4%, which were significantly higher than those of Experiments 1 and 3 (69.5% and 67.2%, respectively), indicating that the efficacy of the composition has a clear concentration dependence.

[0063] The scavenging rates of Experiment Examples 1-3 were significantly higher than those of the monolayer microsphere control group (ROS 58.6-62.1% and DPPH 56.3-59.8%), indicating that the bilayer structure of “inner layer sodium alginate-outer layer chitosan” can synergistically enhance antioxidant efficiency through pH-responsive release.

[0064] The ROS clearance rate (51.2%) and DPPH clearance rate (49.3%) of the mixed free component group (Example 7) were significantly lower than those of Example 3, confirming that microsphere encapsulation can reduce the loss of active ingredients and improve stability.

[0065] The clearance rates of single free components in ratios 3-6 were generally low, with ROS clearance rates ranging from 29.2% to 45.3% and DPPH clearance rates ranging from 27.5% to 42.8%, indicating that single components could not achieve synergistic effects.

[0066] In summary, the pH-responsive bilayer microsphere composition achieves a highly efficient synergistic effect of antioxidant components through a dynamic release mechanism, and its activity is significantly superior to that of monolayer microspheres and free component systems.

[0067] II. Human Efficacy Testing Performance characterization and effect testing results of the products in the examples and comparative examples Test samples: face creams from Examples 1-3, Comparative Examples 1-7, and Blank Example 1 as described in Table 2.

[0068] (2) Subjects: 110 healthy female subjects aged 30-60 years were recruited (10 subjects per group). The inclusion criteria included: healthy women aged 30-60 years, forehead wrinkles grade 3-6, nasolabial folds grade 1-3, facial stratum corneum moisture content ≤45 AU, and transepidermal water loss (TEWL) ≥20 g / (h·m 2 ( ); positive lactic acid sting test (total score ≥3 points). Each group used the same formula product. Written informed consent was obtained. Before enrollment, subjects were asked a series of questions regarding their medical history and health status according to the inclusion and exclusion criteria. Simultaneously, a conformity assessment and skin color test were performed on the test site, and the results were recorded. Environmental conditions: Visual assessment and instrument testing were conducted in an environment with a temperature of 21±1℃ and a relative humidity of 50±10% RH. Visual assessment was conducted under constant lighting conditions (fluorescent tubes or LED lights with a color temperature of 5500~6500K). Subjects were required to adapt to these environmental conditions for at least 30 minutes before assessment and testing could be performed.

[0069] Test method: Directions for use: Apply a sample (0.5 g / time) after cleansing in the morning and evening for 56 consecutive days.

[0070] Testing instruments: Skin wrinkles: C-cube (length, elevation max), Corneometer CM825 (stratum corneum moisture). Skin tone: VISIA-7 (a* value) Test time points: D0 (baseline), D14, D28, D56.

[0071] Table 6 Wrinkle Length Test Data

[0072] Table 7. Wrinkle Elevation Max Test Data

[0073] Table 8. Stratum Corneum Moisture Test Data

[0074] Table 9. Skin Redness a* Value Test Data

[0075] This invention evaluated the anti-aging effects of a pH-responsive anti-aging microsphere composition face cream compared to different carrier types, single-ingredient systems, and mixed free-ingredient systems through a 56-day human efficacy test. The differences between Examples 1-3 were solely in the adjustment of the concentration of active ingredients—Example 2 increased the content of each active ingredient, while Example 3 decreased the content of each active ingredient. Comparative Example 1 consisted of chitosan microspheres encapsulating niacinamide and resveratrol; Comparative Example 2 consisted of sodium alginate microspheres encapsulating berberine and hyaluronic acid; Comparative Examples 3-6 each contained only a single free ingredient: niacinamide, resveratrol, berberine, and hyaluronic acid; and Comparative Example 7 was a mixed system of four free ingredients. The results showed: Regarding wrinkle improvement, Example 2, with its high concentration of active ingredient, showed a wrinkle length change rate of -8.8% within 56 days, significantly better than Example 1 (-6.1%) with a standard concentration and Example 3 (-5.4%) with a low concentration, confirming that efficacy increases with increasing active ingredient concentration. In contrast, Comparative Example 7, with a mixture of four free components, showed a change rate of only -2.0%, Comparative Examples 1-2, with a single carrier encapsulating two components, all showed a change rate of -2.7%, and Comparative Examples 3-6, with a single free component, showed a change rate of only -1.3% to -2.0%, all significantly weaker than Examples 1-3. In terms of wrinkle depth (Elevation max), Example 2 showed a maximum height decrease of 33.2%, Example 1 -27.6%, and Example 3 -16.5%, while Comparative Example 7 only decreased by -10.0%, and Comparative Examples 1-2 decreased by -13.7% to -14.1%, further confirming that the pH-responsive bilayer structure of "inner layer sodium alginate - outer layer chitosan" is superior to single carrier or free component systems.

[0076] In terms of moisturizing and barrier repair, the stratum corneum moisture content of Example 2 increased by 23.8%, which was higher than the 21.9% of Example 1 and the 19.8% of Example 3. In comparison, the comparative example 6 with single free hyaluronic acid increased by only 12.7%, the comparative example 2 with encapsulated hyaluronic acid increased by 15.7%, and the comparative example 7 with mixed free components increased by 15.5%, all of which were lower than that of Example 1, confirming that pH-responsive microspheres can improve long-lasting water retention.

[0077] In terms of anti-inflammatory and soothing effects, the skin redness (a* value) of Example 2 decreased by 18.9%, which was better than -16.7% in Example 1 and -14.4% in Example 3. Comparative Example 7 decreased by -9.7%, Comparative Examples 1-2 decreased by -10.5% to -10.6%, and Comparative Examples 3-6 with single free components decreased by -5.1% to -8.7%, all of which were significantly weaker than Example 1, indicating that the composition has good anti-inflammatory and soothing effects.

[0078] like Figure 1 The image shows facial redness of the subjects before using the face cream described in Example 1, where a is before use (D0), b is after 28 days of use (D28), and c is after 56 days of use (D56). Figure 1 It can be seen that after using the face cream of this invention, with the extension of the usage time, the redness area on the subject's face was significantly reduced and the color was significantly lightened. This is consistent with the skin redness a* value test data of Example 1 in Table 8, indicating that the product of this invention can effectively improve skin erythema and redness and has good anti-inflammatory and soothing effects.

[0079] In summary, this invention achieves a synergistic effect of "daytime anti-oxidation and nighttime repair" by constructing a pH-responsive bilayer microsphere structure of "inner layer sodium alginate and outer layer chitosan". This bilayer microsphere structure can dynamically respond to the diurnal pH changes of the skin: during the day, when the skin surface is neutral (pH 5.5-7.0), the outer chitosan slowly dissolves, gradually releasing niacinamide and resveratrol. Through the synergistic effect of Nrf2 pathway activation and NF-κB inflammation inhibition, it achieves anti-oxidation and daytime protection; at night, when the skin becomes acidic (pH 4.5-5.0), the chitosan layer swells and accelerates the release of niacinamide and resveratrol, exposing the inner sodium alginate microspheres, which release berberine and hyaluronic acid, synergistically enhancing the stratum corneum's water-locking capacity and barrier repair efficiency together with niacinamide and resveratrol. This rhythmic release pattern of "daytime protection - nighttime repair" effectively avoids the activity decay of free ingredients due to pH fluctuations, and solves the technical bottleneck that traditional anti-aging products cannot adapt to the skin's physiological rhythm.

Claims

1. A pH-responsive anti-aging microsphere composition based on regulating circadian rhythms, characterized in that, It consists of an inner layer of sodium alginate microspheres and an outer layer of chitosan coating. The inner layer of sodium alginate microspheres encapsulates berberine and hyaluronic acid. The outer layer of chitosan coating encapsulates nicotinamide and resveratrol with the inner layer of sodium alginate microspheres.

2. The pH-responsive anti-aging microsphere composition based on regulating circadian rhythm according to claim 1, characterized in that, The mass ratio of sodium alginate, berberine, and hyaluronic acid is (1~3):(0.5~1.5):(0.5~1.5). The mass ratio of the inner layer sodium alginate microspheres, nicotinamide, and resveratrol is (5~15):(0.5-1.5):(1.5-2.5).

3. The pH-responsive anti-aging microsphere composition based on regulating circadian rhythm according to claim 1, characterized in that, The inner layer sodium alginate microspheres were prepared by the following method: S11. Add sodium alginate to deionized water and stir until completely dissolved to form a clear solution; S12. Add berberine and hyaluronic acid to the solution obtained in step S11, stir and mix evenly to obtain an aqueous phase; S13. The aqueous phase obtained in step S12 is slowly dripped into the oil phase containing the emulsifier, and a uniform W / O emulsion is formed by high-speed shear emulsification. S14. Add calcium chloride solution dropwise to the emulsion obtained in step S13 and stir to promote cross-linking of sodium alginate molecules into spheres. S15. Centrifuge the product obtained in step S14, collect the precipitated microspheres, wash with deionized water and freeze-dry to obtain inner layer microsphere powder loaded with berberine and hyaluronic acid.

4. The pH-responsive anti-aging microsphere composition based on regulating diurnal rhythm according to claim 3, characterized in that, In step S11, the mass-to-volume ratio of sodium alginate to deionized water is 1~3:100 (w / v), and the stirring conditions are 45~55℃ and 250~350rpm, until it is completely dissolved to form a clear solution; In step S12, the mass concentrations of berberine and hyaluronic acid are 0.5~1.5% w / v and 0.5~1.5% w / v, respectively, and the stirring conditions are: temperature 45~55℃, speed 450~550 rpm, and time 20~40 min. In step S13, the oil phase is liquid paraffin containing 3-6% v / v Span 80, the volume ratio of the water phase to the oil phase is 1:4-6, and the high-speed shear emulsification conditions are a rotation speed of 1200-1600 rpm, a temperature of 35-45℃, and a time of 30-50 min. In step S14, the mass-volume concentration of the calcium chloride solution is 8-12%, and the volume ratio of the calcium chloride solution to the emulsion is 1:4-6. After the addition, the stirring conditions are: temperature 20-30℃, speed 150-250rpm, and time 50-70min. In step S15, the centrifugation conditions are 2500~3500 rpm and the time is 8~12 min; the freeze-drying conditions are -45~-55℃ and the time is 22~26 h.

5. The pH-responsive anti-aging microsphere composition based on regulating diurnal rhythm according to claim 1, characterized in that, The outer chitosan coating layer is prepared by the following method: S21. Dissolve chitosan in an acidic aqueous solution, add nicotinamide and resveratrol, and stir until completely dissolved to obtain a chitosan solution. S22. Add the inner layer sodium alginate microspheres to the chitosan solution obtained in step S21 and disperse them by ultrasonication to form a uniform aqueous phase; S23. Add the aqueous phase obtained in step S22 to the oil phase containing the emulsifier, and stir to emulsify and form a W / O secondary emulsion; S24. Add sodium tripolyphosphate solution dropwise to the secondary emulsion obtained in step S23 and stir to promote cross-linking and solidification of chitosan molecules; S25. Centrifuge the product obtained in step S24, collect the composite microspheres, wash them with anhydrous ethanol and deionized water, and freeze-dry them to obtain pH-responsive sustained-release microsphere solid powder.

6. The pH-responsive anti-aging microsphere composition based on regulating circadian rhythm according to claim 5, characterized in that, In step S21, the acidic aqueous solution is an acetic acid aqueous solution with a volume concentration of 0.5-1.5%, the chitosan dissolved in the acetic acid aqueous solution has a mass-volume concentration of 0.5-1.5% w / v, and the mass concentrations of nicotinamide and resveratrol are 0.5-1.5% w / v and 1.5-2.5% w / v, respectively. The stirring conditions are a temperature of 20-30℃, a rotation speed of 350-450 rpm, and a time of 10-30 min. In step S22, the mass-to-volume ratio of the inner sodium alginate microspheres to the chitosan solution is 5-15% w / v, and the ultrasonic dispersion conditions are 250-350 W power and 5-15 min time, carried out at room temperature. In step S23, the oil phase is liquid paraffin containing 2-4% v / v Tween 80, the volume ratio of the water phase to the oil phase is 1:3-5, and the stirring emulsification conditions are temperature 25-35℃, speed 1000-1500 rpm, and time 25-35 min. In step S24, the mass-volume concentration of sodium tripolyphosphate solution is 0.5-1.5% w / v, and the volume ratio of sodium tripolyphosphate solution to chitosan solution is 1:4-6. After the addition, the stirring conditions are: temperature 20-30℃, speed 550-650 rpm, and time 40-50 min. In step S25, the centrifugation conditions are 3500~4500 rpm for 10~20 min; the freeze-drying conditions are -45~-55℃ for 22~26 h.

7. The application of the pH-responsive anti-aging microsphere composition based on regulating circadian rhythm as described in any one of claims 1-6, characterized in that, The pH-responsive anti-aging microsphere composition is used as an active ingredient in skincare products.

8. The application of the pH-responsive anti-aging microsphere composition based on regulating diurnal rhythm according to claim 7, characterized in that, The skincare products mentioned include toners, serums, lotions, freeze-dried powders, or creams.

9. The application of the pH-responsive anti-aging microsphere composition based on regulating circadian rhythm according to claim 8, characterized in that, The skincare product is a face cream, which comprises phases A, B, C, D, and E, each containing the following components in weight percentage: Phase A includes: Glycerin 5-10%, Butanediol 3-10%, Trehalose 0.5-2%, Sodium hyaluronate 0.01-1%, Tocopheryl acetate 0.01-1%, Panthenol 0.1-5%, Hydroxypropyl methylcellulose 0.1-2%, Sodium acrylate / sodium acryloyldimethyl taurate copolymer 0.1-2%, PEG-100 stearate 0.1-2%, EDTA disodium 0.01-1%, Water balance; Phase B includes: Isopropyl isostearate 0.1-5%, Caprylic / capric triglycerides 0.1-5%, Glyceryl stearate 0.1-5%, Polydimethylsiloxane 0.1-4%, Cetearyl alcohol 0.1-5%, behenol 0.1-2%, Sorbitan oleate 0.1-2%; Phase C includes: Arginine 0.1-2%; Phase D includes: Active ingredient 1-5%; Phase E includes: (Daily use) Fragrance 0.1-2%; Phenoxyethanol 0.01-1%.

10. The application of the pH-responsive anti-aging microsphere composition based on regulating circadian rhythm according to claim 9, characterized in that, The face cream is prepared by the following method: (1) Aqueous phase preparation: Place water in a reactor, fully wet and mix the other components of phase A, add them to the water, stir thoroughly until completely dissolved, and heat to 75-80℃; (2) Preparation of oil phase: Mix the components of phase B and heat to 75-80℃, stirring until completely dissolved; (3) Aqueous phase-oil phase mixing: Slowly pour the oil phase prepared in step (2) into the aqueous phase in step (1), stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min; (4) Add each component in phase C, stir at 250-350 rpm for 3-5 min, then homogenize at 2500-3500 rpm for 3-10 min, and stir to cool to 40-50℃; (5) Add each component in phase D, stir at 250-350 rpm for 3-5 min, and then homogenize at 2500-3500 rpm for 3-10 min; (6) After cooling to 35-38℃, add phase E, stir at 250-350 rpm for 3-10 min, then homogenize at 2500-3500 rpm for 3-10 min to obtain face cream.

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