Composition containing recombinant I-type, III-type and XVII-type humanized collagen as well as preparation method and application of composition

Abstract

The invention is applicable to the field of medicine, and provides a composition containing recombinant I-type, III-type and XVII-type humanized collagen as well as a preparation method and application of the composition, the composition comprises freeze-dried powder and solvent liquid, the freeze-dried powder comprises an A-phase part and a B-phase part, and the solvent liquid comprises an A-phase part and a B-phase part. The solvent liquid comprises a C-phase part and a D-phase part; the compound composition has high purity, good biocompatibility and biological activity, and can be used in the field of scalp care, including but not limited to promotion of hair growth, improvement of scalp health, reduction of alopecia and the like. Meanwhile, the invention further provides a preparation method of the compound composition and application of the compound composition in scalp care and hair growth promotion.

Description

Technical Field

[0001] The present invention belongs to the field of medicine, and in particular relates to a composition containing recombinant humanized type I, type III and type XVII collagen, and a preparation method and use thereof. Background Art

[0002] In modern society, hair loss and scalp health issues are on the rise due to stress, environmental pollution, and unhealthy lifestyles. A healthy scalp is fundamental to hair growth, and improving scalp condition can help prevent hair loss and promote new hair growth. In recent years, collagen has become a focus of research in dermatology and medicine due to its excellent biocompatibility and remarkable efficacy. Collagen excels in maintaining skin health, repairing tissue, and enhancing skin elasticity.

[0003] Type I, III, and XVII collagen are key factors for scalp and hair health, playing important roles in structural support, elasticity maintenance, and cell growth, respectively. Type XVII collagen, in particular, has garnered widespread attention due to its remarkable effects in dermatology and medicine. This collagen plays a crucial role in combating skin aging, promoting hair follicle stem cell regeneration, and preventing hair follicle aging.

[0004] Although type XVII collagen is rare in the human body and difficult to extract, researchers have successfully developed recombinant humanized type XVII collagen that can be produced in large quantities through genetic engineering and recombinant DNA techniques. Experiments and studies have shown that the synergistic effects of recombinant humanized types I, II, and XVII collagens demonstrate significant potential for scalp care and hair regeneration. These different types of recombinant collagen, through their unique functions, collectively promote scalp health and hair growth.

[0005] Many existing products are ineffective in promoting hair growth and improving scalp health. They often require prolonged use for results, and the effects vary widely from person to person. Some scalp care products contain potent ingredients that can cause scalp irritation, allergic reactions, or discomfort, impacting the user experience. Furthermore, the poor water solubility of traditional collagen and other active ingredients results in low effective utilization in products, hindering absorption. Summary of the Invention

[0006] The object of the present invention is to provide a composition containing recombinant humanized collagen types I, III and XVII, aiming to solve the technical problems existing in the prior art identified in the background art.

[0007] The present invention is achieved by providing a composition containing recombinant humanized collagen types I, III and XVII, the composition comprising:

[0008] Lyophilized powder and solvent liquid, wherein the lyophilized powder comprises: a phase A portion and a phase B portion; the solvent liquid comprises: a phase C portion and a phase D portion;

[0009] The phase A specifically comprises: 100 parts of water, 1-10 parts of mannitol, and 1-5 parts of trehalose;

[0010] The phase B specifically comprises: 0.1-2 parts of recombinant humanized type I collagen, 0.1-2 parts of recombinant humanized type III collagen, and 0.1-2 parts of recombinant humanized type XVII collagen;

[0011] The phase C specifically includes: 100 parts of water and 0.1-2 parts of sodium hyaluronate;

[0012] The phase D specifically includes: 0.1-1 part of sodium paraben and 0.1-1 part of sodium dihydrogen phosphate.

[0013] Another object of the present invention is to provide a method for preparing the composition containing recombinant humanized collagen types I, III and XVII as described above, the preparation method comprising the following steps:

[0014] Step 1: Add the water in phase A into the liquid preparation system, and then add mannitol and trehalose in sequence and heat and stir;

[0015] Step 2: Cool the stirred solution and add phase B into the liquid preparation system, stirring until uniform to obtain a solvent solution;

[0016] Step 3: The solvent solution is dispensed into vials through a filling and stoppering machine and half-stoppered;

[0017] Step 4: Transfer the vial to a vacuum freeze dryer for freeze-drying to obtain the freeze-dried powder.

[0018] As a further solution of the present invention, the process of the freeze-drying process is as follows:

[0019] Keep it at 0℃ for 30min, then reduce to -45℃ and keep it at -45℃ for 3h;

[0020] The temperature was raised to -6°C and maintained for 8 h, and the vacuum was maintained, and the temperature was raised to 10°C and maintained for 2 h;

[0021] Keep vacuuming and raise the temperature to 30℃ for 2h;

[0022] Keep vacuuming and raise the temperature to 35°C for 4 h;

[0023] Keep vacuuming to obtain the lyophilized powder.

[0024] As a further solution of the present invention, the heating and stirring in step 1 has a heating temperature of 80°C-90°C and a stirring speed of 400-800 r / min.

[0025] As a further solution of the present invention, the stirred solution is cooled in step 2, and the temperature of the cooled solution is 30°C-40°C.

[0026] Another object of the present invention is to provide a use of the composition containing recombinant humanized collagen types I, III and XVII as described above in scalp care and hair growth promotion.

[0027] The beneficial effects of the present invention are:

[0028] (1) Dosage form advantages: Freeze-drying is a very effective method for drying heat-sensitive products and substances that need to maintain biological activity (collagen). Freeze-dried powder is made by pre-freezing the water in the drug solution using a freeze dryer's vacuum freeze-drying method, and then sublimating the frozen water in the solution under a vacuum and sterile environment to obtain freeze-dried powder. Secondly, the freeze-dried product has a loose shape and basically no color change after drying. After adding a solvent, it can quickly dissolve and restore the physical and chemical properties and biological activity of the original aqueous solution.

[0029] (2) The product is manufactured and filled in a Class 10,000 environment and sterilized by filtering with a microporous membrane, making the product safer and less irritating. Fourth, the moisture content of the product after freeze-drying is very low, which improves the stability of the product and reduces the chance of contamination. This not only facilitates transportation but also extends the shelf life of the product.

[0030] (3) The advantages of recombinant humanized collagen are: 1. Biocompatibility: Recombinant humanized collagen has extremely high biocompatibility because it is designed based on the amino acid sequence of human collagen, which reduces the risk of the human body developing an immune response to it. 2. Low immunogenicity: Because the sequence of recombinant humanized collagen is highly similar to the body's own collagen, it triggers a lower immune response in the human body, which makes it safer in medical applications. 3. No pathogen risk: Compared with traditional animal-derived collagen, the production process of recombinant humanized collagen does not involve animal-derived materials, thereby reducing the risk of pathogen transmission. 3. Water solubility: Recombinant humanized collagen has good water solubility, which makes it more convenient to use in cosmetics and medical products, and also helps to improve its transdermal absorption rate in skin care products. 3. High biological activity: Through genetic engineering technology, recombinant humanized collagen can be optimized to enhance its biological activity, which means that it may be more effective in promoting cell adhesion, proliferation and differentiation.

[0031] (4) The synergistic effect of recombinant humanized collagen complexes has the effects of promoting hair growth, improving scalp health, and reducing hair loss. The synergistic effect of recombinant type XVII collagen, recombinant type I collagen, and recombinant type II collagen has shown great potential in the field of scalp care and hair regeneration. These different types of recombinant collagen promote scalp health and hair growth through their unique functions.

[0032] 1. Recombinant Type XVII Collagen: In scalp tissue, it helps maintain the integrity and elasticity of the scalp and plays a key role in promoting the activity of hair follicle stem cells and regulating the hair growth cycle. It improves the scalp environment by interacting with scalp cells, providing a healthier foundation for hair growth.

[0033] 2. Recombinant Type I Collagen: As the most abundant type of collagen, Type I collagen provides structural support and promotes tissue repair in the scalp, enhancing the strength and elasticity of the scalp, providing a solid foundation for the hair, and reducing hair loss caused by scalp relaxation.

[0034] 3. Recombinant type III collagen: The role of type III collagen in scalp tissue may be related to its role in maintaining tissue moisture and lubrication, which helps to maintain scalp moisture, reduce scalp dryness and irritation, and provide support for hair health. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 A process flow chart provided for an embodiment of the present invention. DETAILED DESCRIPTION

[0036] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0037] A composition containing recombinant humanized collagen types I, III, and XVII. The composition uses a recombinant humanized collagen composition prepared using DNA recombination technology as the primary active ingredient, along with trehalose, a component with moisturizing, antioxidant, and anti-aging properties. Mannitol is used as a lyoprotectant. The solvent comprises purified water, sodium hyaluronate, sodium methylparaben, and sodium dihydrogen phosphate monohydrate. The composition includes:

[0038] Lyophilized powder:

[0039]

[0040] Solvent liquid:

[0041]

[0042] like Figure 1 As shown, a method for preparing a composition containing recombinant humanized collagen types I, III and XVII comprises the following steps:

[0043] Step 1: Add the water in phase A into the liquid preparation system, and then add mannitol and trehalose in sequence, and heat and stir at a temperature of 80°C-90°C at a stirring speed of 400-800 r / min;

[0044] Step 2: Cool the stirred solution to 30-40°C, add phase B into the liquid preparation system, and stir until uniform to obtain a solvent solution;

[0045] Step 3: The solvent solution is dispensed into vials through a filling and stoppering machine and half-stoppered;

[0046] Step 4: Transfer the vial to a vacuum freeze dryer for freeze-drying to obtain the freeze-dried powder.

[0047] In an embodiment of the present invention, the freeze-drying process is as follows:

[0048] Keep it at 0℃ for 30min, then reduce to -45℃ and keep it at -45℃ for 3h;

[0049] The temperature was raised to -6°C and maintained for 8 h, and the vacuum was maintained, and the temperature was raised to 10°C and maintained for 2 h;

[0050] Keep vacuuming and raise the temperature to 30℃ for 2h;

[0051] Keep vacuuming and raise the temperature to 35°C for 4 h;

[0052] Keep vacuuming to obtain the lyophilized powder.

[0053] The recombinant humanized collagen types I, III, and XVII compositions described in this protocol are freeze-dried powders in the form of white, loose, porous blocks. To use, a solvent is simply added dropwise to the freeze-dried powder to reconstitute it into a liquid. This liquid can be applied directly to the scalp or via a microneedle roller. After use, it effectively prevents hair loss and promotes new hair growth.

[0054] Test evaluation:

[0055] 1. Safety assessment test:

[0056] (1) Experimental population:

[0057] 32 subjects aged 18-60 were selected

[0058] (2) Experimental methods:

[0059] Keep your scalp dry or use dry hair. Add the solvent to the freeze-dried powder and reconstitute it. Part the hair and apply an appropriate amount of the reconstituted solution to the scalp. Gently massage until absorbed. No need to rinse. The recommended dosage for topical use is 2-3 ml, and for all-head use is 4-5 ml. Use once daily for four weeks. The specific dosage can be adjusted according to individual needs. Subjects used the product for four consecutive weeks. Any adverse events experienced during product use were observed, questioned, examined, and recorded. The relationship between the adverse events and the product use was determined. The identification criteria are shown in Table 1, and the results are shown in Table 2.

[0060] Table 1: Skin reaction grading standards:

[0061] Skin reactions Grading No response 0 Faint erythema 1 Erythema, infiltration, papules 2 Erythema, infiltration, papules, blisters 3 Erythema, infiltration, bullae 4

[0062] Table 2: Statistics of adverse skin reactions:

[0063] Skin reactions Grading Week 2 Week 4 No response 0 32 cases 32 cases Faint erythema 1 0 cases 0 cases Erythema, infiltration, papules 2 0 cases 0 cases Erythema, infiltration, papules, blisters 3 0 cases 0 cases Erythema, infiltration, bullae 4 0 cases 0 cases

[0064] In summary, during the four-week trial period, a total of 32 subjects used the test product in accordance with regulations. According to the grading criteria for skin reactions in human trials, no adverse skin reactions were observed in any of the subjects throughout the entire testing process.

[0065] 2. Anti-hair loss efficacy test

[0066] This embodiment provides a lyophilized agent and a solvent, which are respectively calculated by total mass as follows:

[0067] Lyophilized agent embodiment:

[0068]

[0069]

[0070] Solvent Example:

[0071] Example Sodium hyaluronate (%) Sodium methylparaben (%) Sodium dihydrogen phosphate (%) water(%) Example 5 0.25 0.1 0.1 Replenishment

[0072] Lyophilized agent + solvent embodiment method:

[0073] Grouping Freeze-dried Solvent Group 1 Example 1 Example 5 Group 2 Example 2 Example 5 Group 3 Example 3 Example 5 Group 4 Example 4 Example 5

[0074] (1) Experimental population:

[0075] 40 subjects aged 25-50 were selected

[0076] (2) Experimental methods:

[0077] This study used the 60-comb method and recruited 80 volunteers aged 25 to 45. They were randomly assigned to four equal groups of 20 participants each. All volunteers had a hair loss count exceeding 20 strands before continuous use of the test sample, and this count remained above 20 strands after a two-week washout period. The volunteers were divided into four groups—Groups 1, 2, 3, and 4—and used the test sample once daily for three months. During the testing period, participants were asked not to use any cosmetics, medications, or devices with hair loss prevention or hair growth properties, and to refrain from hairdressing practices other than haircuts, perms, or dyeing. Hair loss was counted for each volunteer and the data were recorded. The same evaluation and testing was performed on all groups in the first and third months after product use. Detailed test results are shown in Table 3.

[0078] Table 3: Improvement of hair loss in subjects:

[0079]

[0080] It should be understood that, although the various steps in the flow chart of each embodiment of the present invention are shown in sequence according to the indication of the arrows, these steps are not necessarily performed in sequence according to the order indicated by the arrows. Unless otherwise specified herein, the execution of these steps is not strictly limited in order, and these steps can be performed in other orders. Moreover, at least a portion of the steps in each embodiment may include a plurality of sub-steps or a plurality of stages, and these sub-steps or stages are not necessarily performed at the same time, but can be performed at different times, and the execution order of these sub-steps or stages is not necessarily performed in sequence, but can be performed in turn or alternately with at least a portion of other steps or sub-steps or stages of other steps.

[0081] The technical features of the above-mentioned embodiments can be combined arbitrarily. In order to make the description concise, not all possible combinations of the technical features in the above-mentioned embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0082] The above-described embodiments merely illustrate several implementations of the present invention, and while their descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the present invention. It should be noted that a person skilled in the art would be able to make numerous variations and improvements without departing from the spirit of the present invention, all of which fall within the scope of protection of the present invention. Therefore, the scope of protection of the present invention shall be determined by the appended claims.

[0083] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A composition containing recombinant humanized collagen types I, III and XVII, characterized in that: The composition comprises: Lyophilized powder and solvent liquid, wherein the lyophilized powder comprises: a phase A portion and a phase B portion; the solvent liquid comprises: a phase C portion and a phase D portion; The phase A specifically comprises: 100 parts of water, 1-10 parts of mannitol, and 1-5 parts of trehalose; The phase B specifically comprises: 0.1-2 parts of recombinant humanized type I collagen, 0.1-2 parts of recombinant humanized type III collagen, and 0.1-2 parts of recombinant humanized type XVII collagen; The phase C specifically includes: 100 parts of water and 0.1-2 parts of sodium hyaluronate; The phase D specifically includes: 0.1-1 part of sodium paraben and 0.1-1 part of sodium dihydrogen phosphate.

2. A method for preparing the composition containing recombinant humanized collagen types I, III and XVII as claimed in claim 1, comprising the following steps: Step 1: Add the water in phase A into the liquid preparation system, and then add mannitol and trehalose in sequence and heat and stir; Step 2: Cool the stirred solution and add phase B into the liquid preparation system, stirring until uniform to obtain a solvent solution; Step 3: The solvent solution is dispensed into vials through a filling and stoppering machine and half-stoppered; Step 4: Transfer the vial to a vacuum freeze dryer for freeze-drying to obtain the freeze-dried powder.

3. The method according to claim 2, characterized in that The process of the freeze-drying process is as follows: Keep it at 0℃ for 30min, then reduce to -45℃ and keep it at -45℃ for 3h; The temperature was raised to -6°C and maintained for 8 h, and the vacuum was maintained, and the temperature was raised to 10°C and maintained for 2 h; Keep vacuuming and raise the temperature to 30℃ for 2h; Keep vacuuming and raise the temperature to 35°C for 4 h; Keep vacuuming to obtain the lyophilized powder.

4. The method according to claim 3, characterized in that The heating and stirring in step 1 has a heating temperature of 80°C-90°C and a stirring speed of 400-800 r / min.

5. The method according to claim 4, characterized in that The stirred solution is cooled in step 2, wherein the temperature of the cooled solution is 30° C.-40° C.

6. Use of the composition containing recombinant humanized collagen types I, III and XVII as claimed in claim 1 in scalp care and hair growth promotion.

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