Recombinant collagen hair blackening composition based on biomimetic pro-opiomelanocortin peptides and methods of making same

Abstract

This invention relates to the field of oxidative hair dye technology, specifically to a recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides and its preparation method, comprising the following steps: S1: mixing 0.1-25 mg / ml dopamine, 0.5%-5.0% recombinant collagen, and 0.01-2.0 mg / ml biomimetic melanin-promoting peptides to achieve a synergistic effect between the recombinant collagen and the biomimetic melanin-promoting peptides; S2: mixing 30-60 mg / ml sodium periodate with a bio-oxidative enzyme catalyst; S3: dispersing a 15%-30% thermosensitive copolymer in an acetate-sodium acetate buffer solution and controlling the pH of the system to 5.5-7.0; S4: dispensing agent A and agent B separately with the matrix system. This invention highlights the "dual enzymatic synergistic" effect of biomimetic melanin-promoting peptides and polyphenol oxidase, effectively reducing the oxidative activation energy of dopamine.

Description

Technical Field

[0001] This invention relates to the field of hair washing, care and dyeing technology, specifically covering categories such as shampoo, conditioner, hair mask and hair serum, and more specifically to a recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides and its preparation method. Background Technology

[0002] Traditional oxidative hair dyes generally contain carcinogenic substances such as p-phenylenediamine (PPD) and p-aminophenol, and rely on strong alkali (ammonia) and strong oxidants (hydrogen peroxide) to open the hair cuticle, causing permanent damage to the scalp barrier and hair quality.

[0003] Existing technologies, such as patent CN109394558A, although proposing a scheme to use dopamine to mimic melanin, still have the following limitations: 1. The color development time is too long: It usually takes 30-60 minutes to achieve the ideal color depth.

[0004] 2. Lack of hair care function: It only achieves color coverage and cannot repair damaged hair.

[0005] 3. Limited colorfastness: It can withstand approximately 30 washes, and the color may deviate. Summary of the Invention

[0006] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a method for preparing a recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptide, comprising the following steps: S1: mixing 0.1-25 mg / ml of dopamine, 0.5%-5.0% of recombinant collagen and 0.01-2.0 mg / ml of biomimetic melanin-promoting peptide, so that the recombinant collagen and the biomimetic melanin-promoting peptide form a synergistic effect; S2: Mix 30-60 mg / ml of sodium periodate with the biological oxidation enzyme catalyst evenly; S3: Disperse the thermosensitive copolymer at a concentration of 15%-30% in an acetate-sodium acetate buffer solution and control the pH of the system to 5.5-7.0; S4. After combining Agent A and Agent B with the matrix system respectively, the mixture is packaged to obtain a progressive hair darkening composition that undergoes gelation triggered by human skin temperature and a dopamine oxidative polymerization reaction during use.

[0007] Furthermore, in step S1, the recombinant collagen has a triple helix structure; The recombinant collagen utilizes the triple helix structure to form hydrogen bonds with the hair cuticle, thereby filling gaps in the hair strands, enhancing shine, and providing immediate repair for the scalp barrier. The biomimetic melanin-promoting peptide is a specific sequence polypeptide that can significantly reduce the activation energy required for dopamine oxidation. The biomimetic melanin-promoting peptide is used to accelerate the catalytic system in shampooing scenarios, enabling dopamine to complete initial color development within 3-5 minutes and deep color development within 15-20 minutes.

[0008] Furthermore, in step S1, the synergistic effect of the recombinant collagen and the biomimetic melanin-promoting peptide is specifically manifested as follows: While the biomimetic melanin-promoting peptide accelerates the oxidation and polymerization of dopamine to generate melanin-like pigments, the recombinant collagen utilizes its triple helix structure to generate strong hydrogen bond cross-linking with the hair cuticle, clearly exerting the "bio-glue" effect to firmly anchor the generated melanin-like pigment particles to the hair shaft, thereby supporting the high color-fixing stability of the prepared progressive hair blackening composition, which can withstand more than 45 washes. Agent A does not contain p-phenylenediamine or ammonia; Furthermore, through the combination of recombinant collagen, biomimetic melanin-promoting peptides, and dopamine, the final composition forms a specific osmotic pressure difference, which increases the penetration rate deep into the hair follicle by more than 40% compared to ordinary hair care products.

[0009] Furthermore, in step S2, the bio-oxidative enzyme catalyst is polyphenol oxidase; In Agent B, sodium periodate and trace amounts of polyphenol oxidase together constitute an oxidation initiation system.

[0010] Furthermore, the bio-oxidative enzyme catalyst has a synergistic effect with the matrix system; Specifically, when the progressive hair-darkening composition comes into contact with the human scalp, the polyphenol oxidase uses the human skin temperature of about 37°C as a trigger condition, and works with the temperature-sensitive copolymer to cause the composition to undergo an instantaneous transformation from a liquid state to a gel state (i.e., in-situ gelation process), which is used to lock in the active ingredients and prevent the composition from dripping or running during use as a core technical means.

[0011] Further, in step S3, the thermosensitive copolymer includes a Prönnik thermosensitive matrix; The thermosensitive copolymer is one or a mixture of two of F127 and F68.

[0012] Furthermore, in step S3, the acetate-sodium acetate buffer solution controls the pH of the system within the range of 5.5-7.0; The specified pH range ensures that the matrix system is in a weakly acidic range of 5.5-7.0. This, combined with agent A, strictly controls the system to be free of p-phenylenediamine and ammonia, while providing a suitable physiological environment for the oxidative polymerization of dopamine and the repair function of the recombinant collagen.

[0013] Furthermore, in step S4, the packaging adopts a dual-chamber or multi-chamber isolation packaging form; After preparation, Agent A and Agent B are stored in a mutually isolated state within the packaging. When used, they are immediately mixed by squeezing or pumping to initiate the oxidative polymerization reaction of dopamine in the composition.

[0014] Furthermore, the progressive hair-darkening composition obtained after the combination molding is not limited to oxidative hair dyes, and its specific product forms include any one of shampoo, conditioner, hair mask and hair essence; when the progressive hair-darkening composition is used, the material forms a gel state under the trigger of human skin temperature to prevent dripping, and at the same time, the dopamine, under the accelerated catalysis of the biomimetic melanin-promoting peptide, completes the initial color development within 3-5 minutes and completes the deep coloring within 15-20 minutes.

[0015] The recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides is prepared by the method described in any one of claims 1 to 9.

[0016] Beneficial effects This invention highlights the "dual enzymatic synergistic" effect of biomimetic melanin-promoting peptides and polyphenol oxidase, effectively reducing the oxidative activation energy of dopamine. This is the key to achieving initial color development within 3-5 minutes and deep coloring within 15-20 minutes, and is also the core technological barrier that distinguishes this composition from ordinary dopamine products on the market. The introduction of recombinant collagen, with its triple helix structure, forms hydrogen bonds with hair strands, filling the damaged gaps in the hair while firmly anchoring the generated melanin to the hair shaft, increasing the hair's wash resistance to more than 45 times, and increasing the penetration rate of the core components into the hair follicle by more than 40% compared to conventional products. In addition, combined with the temperature-sensitive copolymer (F127 / F68) system, the material undergoes an "in-situ gelation" process from liquid to gel state when in contact with a human skin temperature of 37°C. This rheological phase change property serves as the core technology, solving the industry pain point of traditional shampoos and other hair care products easily "dripping" during use. It is strictly labeled as free of p-phenylenediamine (PPD) and ammonia, and the system pH value is in the weakly acidic range of 5.5-7.0. While ensuring color development efficiency and color fixation durability, it achieves safe and gentle repair of the scalp barrier, perfectly meeting the brand positioning and high standards of "medical-grade safety". Attached Figure Description

[0017] Figure 1 This is a flowchart illustrating the preparation method of the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to the present invention. Detailed Implementation

[0018] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0019] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but includes other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0020] The present invention will now be described in further detail with reference to the accompanying drawings: Example: like Figure 1 As shown, the preparation method of the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides includes the following steps: S1: Mix 0.1-25 mg / ml of dopamine, 0.5%-5.0% of recombinant collagen with 0.01-2.0 mg / ml of biomimetic melanin-promoting peptide to achieve a synergistic effect between the recombinant collagen and the biomimetic melanin-promoting peptide. S2: Mix 30-60 mg / ml of sodium periodate with the biological oxidation enzyme catalyst evenly; S3: Disperse the thermosensitive copolymer at a concentration of 15%-30% in an acetate-sodium acetate buffer solution and control the pH of the system to 5.5-7.0; S4. After combining Agent A and Agent B with the matrix system respectively, they are packaged to obtain a progressive hair darkening composition that undergoes gelation triggered by human skin temperature and a dopamine oxidative polymerization reaction during use.

[0021] Furthermore, the specific implementation process of step S1 is as follows: Agent A is formulated as the biomimetic melanin precursor and structural repair center of the entire progressive hair darkening composition. Its specific component ratio, microscopic mechanism of action, and formulation process play a decisive role in the color development efficiency and color fixation durability of the final product.

[0022] Specifically, the preparation of Agent A involves thoroughly mixing dopamine (0.1-25 mg / ml), recombinant collagen (0.5%-5.0% by mass), and biomimetic melanocyte-stimulating peptide (0.01-2.0 mg / ml) in purified water at room temperature to mild heating (e.g., 25℃-40℃) until a homogeneous, transparent or semi-transparent liquid system is formed. During this preparation process, strict control is maintained to prevent the addition of highly irritating and potentially carcinogenic chemicals such as p-phenylenediamine (PPD) and ammonia.

[0023] First, the biomimetic melanin-promoting peptide with a specific sequence introduced into the system acts as a highly efficient biocatalytic medium. Its main function is to significantly reduce the activation energy required for dopamine to undergo subsequent oxidative polymerization. Through the targeted catalysis of this melanin-promoting peptide, the technical bottleneck of traditional dopamine-based melanin-promoting technology, which typically requires 30-60 minutes for color development, is overcome. This allows the composition of the present invention to achieve initial color development within 3-5 minutes and rapid deep color development within 15-20 minutes in short-contact scenarios such as shampooing, with a color development efficiency 3 to 4 times higher than that of existing technologies.

[0024] Agent A innovatively incorporates recombinant collagen, a recombinant human-like collagen with a naturally stable triple helix structure. In the dynamic process of dopamine being accelerated by melanocyte-stimulating peptides to generate melanin-like molecules, the recombinant collagen exerts a unique "bio-glue" effect.

[0025] On the one hand, collagen uses its triple helix structure to form a dense hydrogen bond cross-linking network with the hair cuticle, which can not only target and fill the gaps in the hair caused by daily damage, significantly improving the shine of the hair (i.e., increasing the L value in colorimetry), but also play an immediate repair role on the damaged scalp barrier of consumers.

[0026] On the other hand, this hydrogen bond cross-linking network based on the triple helix structure acts like a high-strength "bioadhesive" or "bioglue," firmly anchoring and locking the newly generated melanin-like particles into the gaps in the hair cuticle, thereby greatly improving the color fixation stability. This significantly increases the number of washes the treated hair can withstand, from the traditional 30 times to more than 45 times, providing solid data support.

[0027] By scientifically proportioning dopamine, recombinant collagen, and biomimetic melanin-promoting peptides in step S1, a specific bio-osmotic pressure difference is created in solution for agent A. Experimental data shows that, driven by this osmotic pressure difference, the penetration rate of the core active ingredients in the composition of this invention into the depths of the hair follicle and the interior of the hair cuticle is increased by more than 40% compared to commercially available ordinary shampoos or regular hair care products, further solidifying the gradual hair-darkening effect from the surface to the core.

[0028] To enable those skilled in the art to more clearly implement step S1, three specific formulation examples of Agent A are provided below to support the parameter range in the claims: Example 1 of preparation of agent A (preferred optimal ratio): Add an appropriate amount of deionized water to the reaction vessel, and then accurately weigh and add the following components in sequence: dopamine 10.0 mg / ml, recombinant collagen 2.5% (w / v), and biomimetic melanin-stimulating peptide 0.5 mg / ml. Turn on the homogenizer and stir continuously at 300-500 rpm for 20 minutes at 30°C until the system is completely dissolved and homogeneous, obtaining the optimal performance of the core component solution of Agent A. The penetration rate and synergistic color-fixing effect reach their peak at this formulation.

[0029] Example 2 of Agent A preparation (low-concentration daily maintenance ratio): Take deionized water and add the following in sequence: dopamine 2.0 mg / ml, recombinant collagen 0.8% (w / v), and biomimetic melanin-promoting peptide 0.05 mg / ml. Stir and mix at room temperature (25°C) for 30 minutes. This formulation is extremely mild and suitable for preparing daily, gradual hair darkening shampoo and conditioner for frequent use.

[0030] Example 3 of Agent A preparation (high-concentration, rapid, deep-dyeing formulation): Take deionized water and add the following in sequence: dopamine 22.0 mg / ml, recombinant collagen 4.5% (w / v), and biomimetic melanin-promoting peptide 1.8 mg / ml. Mix at a constant temperature of 35°C with stirring for 15 minutes. In this example, the high concentration of dopamine combined with melanin-promoting peptide is specifically designed for scenarios requiring rapid, deep coloring and where hair quality is severely damaged. The high concentration of collagen provides ultimate "bio-glue" encapsulation and repair capabilities.

[0031] Furthermore, the specific implementation process of step S2 is as follows: After completing the preparation of the core component liquid, step S2 of the composition preparation method of the present invention involves constructing a matching oxidation initiation system (Agent B). This system constitutes the kinetic core of the entire composition in the practical application stage, and its formulation design not only determines the start-up rate of dopamine oxidative polymerization, but also directly affects the physical morphological changes of the product during use.

[0032] In the specific preparation process, the preparation of agent B requires the precise compounding of sodium periodate with trace amounts of polyphenol oxidase, with the concentration strictly controlled within the range of 30-60 mg / ml. Considering the natural sensitivity of polyphenol oxidase as a bioactive polymer to temperature and shear force, in practice, both are usually uniformly dispersed in a suitable solvent carrier using a low-shear stirring device under low temperature (preferably 4℃-15℃) or controlled room temperature conditions, thereby maximizing the preservation of the enzyme's catalytic activity.

[0033] The technological breakthrough of this oxidation initiation system lies in its complete departure from the path dependence of traditional single chemical oxidation. Sodium periodate in the system acts as the primary oxidant, providing the necessary basic oxidation potential for the subsequent activation of dopamine polymerization in agent A. However, simple high-concentration chemical oxidation often carries the risk of uncontrollable reactions or oxidative stress damage to the scalp. Therefore, this invention innovatively introduces polyphenol oxidase into agent B, endowing the system with unique environmental response characteristics.

[0034] The optimal catalytic temperature range of polyphenol oxidase highly overlaps with the normal scalp temperature (approximately 37°C). When the dispensed and stored agents A and B are mixed and applied to the hair and scalp by the consumer, the slight increase in ambient temperature can instantly trigger the catalytic peak of polyphenol oxidase. In this biochemical reaction process, enzymatic catalysis not only accelerates the conversion of dopamine into melanin precursors many times over, but the changes in the intermolecular interactions caused by the reaction also constitute a thermodynamic response triggering mechanism.

[0035] This microscopic change, driven by both temperature and enzymatic reactions, directly induces rapid hydrophobic association of the subsequently bound thermosensitive copolymer segments within the system. Macroscopically, this manifests as a rapid rheological phase transition within a very short time of contact with human skin, transforming the originally highly fluid liquid composition into a gel state with a certain degree of adhesion. This in-situ gelation process completely overcomes the industry-wide technical challenge of traditional water-based or emulsion-based shampoos and conditioners easily sagging during their contact with the skin. More importantly, the formed gel network precisely anchors highly active melanin-mimicking precursors and recombinant collagen to the hair shaft surface and cuticle crevices, preventing the loss of effective ingredients and ensuring the simultaneous completion of deep coloring and repair effects.

[0036] To further clarify the implementation details of Agent B, the following are three preparation examples with different set parameters: Example 1 of preparation of agent B (preferred scheme for balanced color development): In a temperature-controlled reactor at 10℃, add an appropriate amount of purified water, accurately weigh and add sodium periodate with a concentration of 45.0 mg / ml. After it is completely dissolved, slowly add polyphenol oxidase with a purity greater than 95% (addition amount: 0.05 mg / ml). Turn on the paddle stirrer and gently stir at a low speed of 100 rpm for 15 minutes. The oxidation potential and enzymatic activity at this ratio reach the optimal balance, providing the most stable 37℃ in-situ gelation experience when used in conjunction with Agent A, suitable for the general gradual blackening needs of most hair types.

[0037] Example 2 of preparation of agent B (mild formulation): Under controlled room temperature (approximately 20°C), sodium periodate at a concentration of 30.0 mg / ml and polyphenol oxidase at a concentration of 0.02 mg / ml were dispersed in a solvent and stirred continuously for 20 minutes. This method utilizes the lower limit of the oxidant concentration within the scope of the claims and is specifically designed for sensitive scalps and severely damaged hair. It relies on the highly efficient catalysis of polyphenol oxidase to compensate for the reduction in the concentration of chemical oxidants, achieving basic color development and fixation goals while ensuring safety and non-irritation.

[0038] Example 3 of Agent B Formulation (High-Efficiency Deep Dyeing Solution): A mixture containing 60.0 mg / ml sodium periodate and 0.1 mg / ml polyphenol oxidase was prepared under ice-water bath protection at 5°C. This high-concentration formulation is designed to address coarse, resistant hair or applications requiring rapid, deep coloring. Due to the increased enzyme concentration, the phase transition rate upon contact with the scalp is faster, resulting in a stronger gel-locking layer that prevents the high-concentration reactants from flowing down and staining the facial skin.

[0039] Furthermore, the specific implementation process of step S3 is as follows: As one of the core frameworks of the composition preparation method of this invention, step S3 aims to construct a matrix system that combines rheological intelligent response and high physiological compatibility. This matrix system not only provides a spatial framework for the dopamine oxidative polymerization and recombinant collagen penetration in agent A, but also serves as the material basis for realizing the "in-situ gelation" anti-sagging mechanism in step S2.

[0040] In the specific preparation process, the matrix system is formulated strictly according to the "cold dispersion" principle. During operation, an acetate-sodium acetate buffer solution with a pH of 5.5-7.0 needs to be prepared in advance and cooled to an ice-water bath environment of 0℃-5℃. Subsequently, under continuous and slow shear force, a thermosensitive copolymer with a mass-to-volume ratio (w / v) of 15%-30% is added in batches at a uniform rate to the aforementioned low-temperature buffer solution, and allowed to swell for 12 to 24 hours until the polymer chains are fully extended and a clear, transparent, highly fluid solution is formed.

[0041] Selecting a specific concentration of thermosensitive copolymer (preferably a Pluronic series amphiphilic triblock copolymer, such as Poloxamer 407 / F127 and Poloxamer 188 / F68, used alone or in combination) is key to achieving the thermodynamic phase transition of the composition of this invention. From a microscopic molecular dynamics perspective, the Pluronic copolymer is composed of hydrophilic polyethylene oxide (PEO) and hydrophobic polypropylene oxide (PPO) blocks. At room temperature or lower, both PEO and PPO blocks can form stable hydrogen bonds with water molecules, making the system exhibit a low-viscosity liquid state (Sol state). However, when the system comes into contact with the human scalp (approximately 37°C), and the ambient temperature exceeds its critical micelle temperature (CMT), the hydrophobic PPO blocks undergo intense dehydration, driving the polymer monomers to self-assemble into nanoscale micelles with PPO as the core and PEO as the shell. As the local micelle concentration surges, the micelles become tightly packed and entangled. The entropy-driven force of the system causes it to undergo a rheological abrupt change from a liquid state to a high-viscosity gel state in a very short time. This mechanism perfectly complements the temperature-responsive characteristics of polyphenol oxidase, constructing a spatial network physical barrier upon contact with hair, completely blocking the path of traditional hair dyes that drip or drip.

[0042] Meanwhile, the precise pH setting in step S3 (acetic acid-sodium acetate buffer system of 5.5-7.0) has multiple and irreplaceable technical benefits. Physiologically, this weakly acidic to neutral range closely matches the natural "weakly acidic sebum film" environment of the human scalp. Combined with the absence of p-phenylenediamine and ammonia in agent A, the chemical irritation to the scalp barrier is minimized.

[0043] At the level of chemical reaction kinetics, the oxidative polymerization rate of dopamine is extremely sensitive to the proton (H+) concentration of the system. This buffer solution provides excellent resistance to acid-base interference, enabling dopamine to generate melanin precursors at a stable and controllable rate under the synergistic catalysis of melanopeptide and polyphenol oxidase. Furthermore, the suitable pH environment effectively prevents thermal denaturation or acid-base depolymerization of the triple helix conformation of recombinant collagen, ensuring its maximum activity as a structural repair agent. To further illustrate the formulation parameters and rheological properties of the matrix system, three typical examples of matrix system preparation are listed below: Matrix system preparation example 1 (preferred monomeric thermosensitive formulation): A 0.1 mol / L acetate-sodium acetate buffer solution was prepared, and the pH of the system was precisely calibrated to 6.0 by adjusting the molar ratio of acetic acid to sodium acetate. This buffer solution was placed in a 4°C freezer, and 20% (w / v) of Pluronic F127 powder was slowly added, with gentle stirring to ensure uniform dispersion. The solution was then allowed to swell completely in a cold environment for 24 hours. This embodiment exhibits a phase transition temperature (Tgel) of approximately 32-34°C, excellent fluidity at room temperature, and easy mixing and dispensing with agents A and B. Upon contact with the scalp, it forms a gel rapidly with moderate gel strength, making it the optimal matrix framework for commercially available progressive hair darkening shampoos.

[0044] Example 2 of matrix system preparation (two-component compound anti-dehydration formulation): At 4°C, 18% (w / v) of Pranic F127 and 4% (w / v) of Pranic F68 were added to an acetate-sodium acetate buffer solution with a pH of 5.5. The introduction of F68 not only acted as a co-gelling agent to adjust the critical micelle concentration of the system, but also, due to its excellent surface activity, further enhanced the wetting and spreading ability of the composition on the hair dryer surface. This method resulted in a gel with higher porosity after the phase transition, which facilitated the continuous penetration of high-concentration recombinant collagen and melanin-mimicking molecules into the deeper layers of the hair cuticle.

[0045] Matrix system preparation example 3 (high-strength retention formulation): A slightly acidic buffer solution with a pH of 6.8 was prepared, and Pluronic F127 was dispersed and swollen to a concentration of 28% (w / v) in an ice-water bath. This formulation is close to the upper limit of the scope of protection of the claims, and its polymer solid content is extremely high. This matrix system is specifically designed for targeted application to areas of severe gray hair (such as the temples). It exhibits extremely high gel modulus (G') in response to body temperature, resulting in extremely strong adhesion and ensuring ultra-long-lasting high-concentration retention of the active ingredient in the localized area.

[0046] Furthermore, the specific implementation process of step S4 is as follows: After completing the formulation of Agent A, Agent B, and the matrix system, proceed to step S4. This step is not only an engineering step for achieving commercialization of the product, but also a key structural design to ensure that the highly active biomimetic blackening precursor and oxidation initiation system maintain thermodynamic stability during storage and achieve kinetic activation upon use.

[0047] In actual filling and assembly processes, the matrix system is typically premixed into agent A and agent B under homogeneous conditions in predetermined proportions (e.g., equal volume or equal mass ratio) to form phase A and phase B materials with non-Newtonian fluid characteristics. Subsequently, the two phases A and B are packaged in dual-chamber or multi-chamber isolation containers. The initial design purpose of this spatial isolation is to establish a physical barrier, completely cutting off the electron transfer path between dopamine monomers and sodium periodate / polyphenol oxidase during storage, preventing premature consumption of effective oxidation potential and ineffective self-polymerization of the active ingredient.

[0048] When the user operates the packaging container (e.g., by pressing the linkage pump head), the A and B phase materials are extruded synchronously at a set specific volume ratio (preferably 1:1) and instantaneously mixed in situ within the flow channel or between the palms. Under the physical shear force of extrusion and kneading, the material exhibits excellent shear thinning characteristics, enabling it to spread quickly and evenly on the surface of hair and scalp.

[0049] More importantly, once the mixed composition comes into contact with human skin, the cascade reaction of the entire system is immediately initiated. At the macroscopic rheological level, as described in step S3 above, the material undergoes a thermo-induced phase transition from a liquid to a gel state triggered by a skin temperature of approximately 37°C. The in-situ constructed three-dimensional gel network precisely locks the reactants to the target area. At the microscopic reaction kinetic level, the activity of polyphenol oxidase is instantly activated by body temperature, synergistically reducing the activation energy threshold of dopamine oxidation polymerization in two ways, in conjunction with the biomimetic melanin-promoting peptide in Agent A. This microenvironment, constructed through the synergistic effect of physical phase transition and bioenzyme, causes an exponential increase in the oxidation rate of dopamine: within 3-5 minutes after use, dopamine completes a rapid conversion to intermediate products such as dihydroxyindole, macroscopically manifested as the initial color development of hair; during the 15-20 minute residence period, the intermediate products further polymerize and cross-link to form a large molecular melanin-mimicking network, while recombinant collagen is anchored to the hair cuticle through hydrogen bonds and fills structural voids.

[0050] To demonstrate the above-mentioned molding process and its application in progressive hair darkening shampoo and conditioner products, the following are examples of three typical product forms: Product Portfolio Example 1 (Daily Gradual Darkening Shampoo): The A agent prepared in Example 2 was mixed with an appropriate amount of matrix from Example 1 to form phase A, and the B agent prepared in Example 2 was mixed with an equal amount of matrix from Example 1 to form phase B. The two phases were separately filled into standard dual-chamber pump bottles made of PETG material, with the pump core displacement ratio set at 1:1. The shampoo prepared in this example has a shelf life of over 36 months at room temperature. Consumers can obtain an equal amount of the mixture simply by pressing the pump head during daily shampooing, without the need for additional mixing. The mixture foams rapidly upon application and undergoes a gel phase transition, achieving initial coloring and hair stabilization within the typical 3-5 minute shampooing time, perfectly suited to the fast-paced modern lifestyle.

[0051] Product Combination Example 2 (Intensive Repair Hair Darkening Mask): Example 3 (Agent A) and Example 3 (Agent B) were combined with Example 3 (High-Strength Residual Matrix). To prevent the oxidative inactivation of high-concentration active enzymes and dopamine, the preferred packaging is a vacuum-sealed double-tube soft bag structure. This structure ensures zero contact between the product and external oxygen throughout the entire usage cycle. Due to the use of a high-concentration system and a high-strength gel matrix, this example, after being squeezed and applied to severely damaged or locally heavily gray hair areas, even after leaving it for 15-20 minutes for deep coloring, poses absolutely no risk of product dripping or running and staining the facial skin, ultimately providing professional salon-grade blackening and hair core reshaping effects.

[0052] Product Portfolio Example 3 (Microchannel Instant Mix Hair Serum): Example 1 using Agent A and Example 1 using Agent B are combined with Example 2, a two-component compound anti-dehydration matrix. The packaging design employs a specially designed dual-chamber flow-guiding comb container with a microchannel static mixer. The material undergoes high-intensity turbulent mixing within micron-level channels upon being extruded from the comb teeth, and is then directly applied to the hair roots and follicles. This example fully utilizes the system's excellent osmotic pressure difference, further maximizing its penetration deep into the hair follicles, making it particularly suitable for precise, gradual application to the roots of newly emerging gray hair.

[0053] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides, characterized in that, Includes the following steps: S1: Mix 0.1-25 mg / ml of dopamine, 0.5%-5.0% of recombinant collagen and 0.01-2.0 mg / ml of biomimetic melanin-promoting peptide to achieve a synergistic effect between the recombinant collagen and the biomimetic melanin-promoting peptide, and prepare Agent A. S2: Mix 30-60 mg / ml of sodium periodate with a biological oxidation enzyme catalyst to prepare agent B; S3: Disperse the thermosensitive copolymer at a concentration of 15%-30% in an acetate-sodium acetate buffer solution and control the pH of the system to 5.5-7.0 to construct the matrix system; S4. After combining Agent A and Agent B with the matrix system respectively, the mixture is packaged to obtain a progressive hair darkening composition that undergoes gelation triggered by human skin temperature and a dopamine oxidative polymerization reaction during use.

2. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 1, characterized in that, In step S1, the recombinant collagen has a triple helix structure; The recombinant collagen utilizes the triple helix structure to form hydrogen bonds with the hair cuticle, thereby filling gaps in the hair strands, enhancing shine, and providing immediate repair for the scalp barrier. The biomimetic melanin-promoting peptide is a specific sequence polypeptide that can significantly reduce the activation energy required for dopamine oxidation. The biomimetic melanin-promoting peptide is used to accelerate the catalytic system in shampooing scenarios, enabling dopamine to complete initial color development within 3-5 minutes and deep color development within 15-20 minutes.

3. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 2, characterized in that, In step S1, the synergistic effect of the recombinant collagen and the biomimetic melanin-promoting peptide is specifically manifested as follows: While the biomimetic melanin-promoting peptide accelerates the oxidation and polymerization of dopamine to generate melanin-like pigments, the recombinant collagen utilizes its triple helix structure to generate strong hydrogen bond cross-linking with the hair cuticle, clearly exerting the "bio-glue" effect to firmly anchor the generated melanin-like pigment particles to the hair shaft, thereby supporting the high color-fixing stability of the prepared progressive hair blackening composition, which can withstand more than 45 washes. Agent A does not contain p-phenylenediamine or ammonia; Furthermore, through the combination of recombinant collagen, biomimetic melanin-promoting peptides, and dopamine, the final composition forms a specific osmotic pressure difference, which increases the penetration rate deep into the hair follicle by more than 40% compared to ordinary hair care products.

4. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 3, characterized in that, In step S2, the bio-oxidation enzyme catalyst is polyphenol oxidase; In Agent B, sodium periodate and trace amounts of polyphenol oxidase together constitute an oxidation initiation system.

5. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 4, characterized in that, The bio-oxidative enzyme catalyst has a synergistic effect with the matrix system; Specifically, when the progressive hair-darkening composition comes into contact with the human scalp, the polyphenol oxidase uses the human skin temperature of about 37°C as a trigger condition, and works with the temperature-sensitive copolymer to cause the composition to undergo an instantaneous transformation from a liquid state to a gel state, which is used to lock in the active ingredients and prevent the composition from dripping or running during use.

6. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 5, characterized in that, In step S3, the thermosensitive copolymer includes a Prönnik thermosensitive matrix; The thermosensitive copolymer is one or a mixture of two of F127 and F68.

7. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 6, characterized in that, In step S3, the acetate-sodium acetate buffer solution controls the pH of the system within the range of 5.5-7.0; The specified pH range ensures that the matrix system is in a weakly acidic range of 5.5-7.

0. This, combined with agent A, strictly controls the system to be free of p-phenylenediamine (PPD) and ammonia, while providing a suitable physiological environment for the oxidative polymerization of dopamine and the repair function of the recombinant collagen.

8. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 7, characterized in that, In step S4, the packaging adopts a dual-chamber or multi-chamber isolation packaging method; After preparation, Agent A and Agent B are stored in a mutually isolated state within the packaging. When used, they are immediately mixed by squeezing or pumping to initiate the oxidative polymerization reaction of dopamine in the composition.

9. The method for preparing the recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides according to claim 8, characterized in that, The progressive hair-darkening composition obtained after the combination molding is not limited to oxidative hair dyes. Its specific product forms include any one of shampoo, conditioner, hair mask, and hair essence. When the progressive hair-darkening composition is used, the material forms a gel state under the trigger of human skin temperature to prevent dripping. At the same time, the dopamine, under the accelerated catalysis of the biomimetic melanin-promoting peptide, completes the initial color development within 3-5 minutes and completes the deep coloring within 15-20 minutes.

10. A recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides, characterized in that, The hair-darkening composition is prepared by the method described in any one of claims 1 to 9 for preparing a recombinant collagen hair-darkening composition based on biomimetic melanin-promoting peptides.

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