In-situ spray polyamino acid hydrogel as well as preparation method and application thereof

Abstract

The invention relates to in-situ spray polyamino acid hydrogel as well as a preparation method and application thereof, and belongs to the technical fields of biological medicines, daily chemicals and the like. The preparation method comprises the following steps: dissolving polyglutamic acid and polyaspartic acid to prepare a polyamino acid polymer aqueous solution, arginine, lysine, diaminopyrimidine oxide, pyrrolidinyl diaminopyrimidine oxide and natural plant essential oil; the plant extracellular vesicles are dissolved and dispersed in an aqueous solution to prepare a composite solution, and the carboxyl coupling agent is added to form the self-adaptive sprayable polyamino acid hydrogel. The preparation method is green and environment-friendly, no toxic or harmful solvent is generated, other chemical cross-linking agents are not introduced in the hydrogel cross-linking process, the used raw materials are safe and environment-friendly, and the biosafety is very high. The sprayable polyamino acid hydrogel formed after crosslinking causes excellent gelation performance and hair follicle affinity and can be widely applied to the fields of tissue engineering, regenerative medicine and daily cosmetics.

Description

Technical Field

[0001] The invention relates to an in-situ spray polyamino acid hydrogel and a preparation method and application thereof, belonging to the technical fields of biomedicine, daily chemicals and the like. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] As the pace of society and people's lives continues to accelerate, hair loss is becoming more common and is gradually showing a trend of younger people. Hair loss is related to many factors such as genetic factors, living habits, endocrine, mental state, certain diseases, etc. Hair loss not only affects the appearance of people with hair loss, but also easily causes people psychological pressure and seriously affects their quality of life.

[0004] Currently, commonly used treatments for hair loss include drug therapy, surgical transplantation, high-intensity laser therapy, and natural plant extracts as active ingredients in cosmetics. Traditional drug treatments are mainly based on minoxidil and finasteride, but the treatment process can cause side effects such as obesity, sexual dysfunction, and depression that cannot be ignored. Surgical transplantation is expensive, requires doctors to have skilled operating techniques, and the survival rate of hair follicles after transplantation is limited. Low-energy laser treatment takes a long time and may cause local skin symptoms such as dry skin, itching, redness, and tight scalp. Natural plant extracts have problems such as poor stability of plant-derived extracts, easy decomposition, complex ingredients, unclear effective substances and content, unclear mechanism of action, and long effective time. Therefore, it is of great significance to develop a safe and effective anti-hair loss raw material to solve the problem of hair loss. Summary of the invention

[0005] In order to address the deficiencies of the prior art, the purpose of the present invention is to provide an in-situ spray polyamino acid hydrogel and a preparation method and application thereof. The present invention utilizes the advantages of high water content, adaptability, and biocompatibility of the hydrogel to reshape the 3D cell dynamic microenvironment of damaged scalp tissue, improve the vitality of hair follicle cells, and prolong the growth period of hair follicles, thereby playing a role in preventing hair loss and regenerating hair.

[0006] In order to achieve the above object, the technical solution of the present invention is:

[0007] In a first aspect, a method for preparing an in-situ spray polyamino acid hydrogel comprises the following steps:

[0008] dissolving polyglutamic acid and polyaspartic acid to prepare a polyamino acid aqueous solution;

[0009] Arginine, lysine, diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oil, and plant extracellular vesicles are dissolved and dispersed uniformly to prepare a composite solution;

[0010] providing a carboxyl activator solution;

[0011] When in use, the polyamino acid aqueous solution, the composite solution and the carboxyl activator solution are sprayed and mixed.

[0012] The polyamino acid aqueous solution and composite solution prepared by the present invention can be cross-linked in situ to form an adaptive sprayable polyamino acid hydrogel after adding a carboxyl activator. The sprayable polyamino acid hydrogel formed after cross-linking has excellent gelation performance and hair follicle affinity. Since the polyamino acid hydrogel prepared by the present invention has a fast response speed and is easy to agglomerate in ordinary mixing, spray mixing is required to form a uniform hydrogel.

[0013] In some embodiments, the mass concentration of polyglutamic acid in the polyamino acid aqueous solution is 2-20wt%, and the mass ratio of polyglutamic acid to polyaspartic acid is (5-30):1.

[0014] In some embodiments, the solvent for dissolving polyglutamic acid and polyaspartic acid includes but is not limited to at least one of MES buffer, Tris buffer and ultrapure water. The pH value of MES buffer and Tris buffer is 4.0-8.0.

[0015] In some embodiments, arginine and lysine are dissolved to form an amino acid composite solution; after diaminopyrimidine oxide and pyrrolidinyl diaminopyrimidine oxide are dissolved, the amino acid composite solution is added to dissolve, and then natural plant essential oils and plant extracellular vesicles are added to mix evenly. Specifically, the amino acid composite solution is added in a non-one-time addition manner (e.g., continuous addition, addition in batches, dropwise addition, etc.). Specifically, the solvent for dissolving arginine and lysine is water. Specifically, the solvent for dissolving diaminopyrimidine oxide and pyrrolidinyl diaminopyrimidine oxide is an alcohol solvent. The alcohol solvent of the present invention is, for example, methanol, ethanol, isopropanol, ethylene glycol, glycerol (glycerol), etc., preferably a mixture of ethanol and glycerol.

[0016] In some embodiments, the mass concentration of arginine in the composite solution is 1-10wt%, the mass ratio of arginine to lysine is 1:(4-20), and the mass ratio of arginine to diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oil, and plant extracellular vesicles is 1:(0.01-0.1):(0.01-0.1):(0.05-0.5):(0.001-0.5).

[0017] In some embodiments, the natural plants in the natural plant essential oil include, but are not limited to, at least one of ginger root, codonopsis pilosula, mint, angelica sinensis, and Millettia reticulata.

[0018] In some embodiments, the plant in the plant extracellular vesicles includes but is not limited to at least one of ginseng, aloe vera, Polygonum multiflorum, and Platycladus orientalis leaves.

[0019] In the preparation method of the present invention, the polyamino acid aqueous solution, the composite solution and the carboxyl activator solution can be directly sprayed for mixing; the carboxyl activator solution can also be added to the polyamino acid aqueous solution to prepare a precursor solution A, and when used, the precursor solution A and the composite solution are sprayed for mixing; the carboxyl activator solution can also be added to the composite solution to prepare a precursor solution B, and when used, the polyamino acid aqueous solution and the precursor solution B are sprayed for mixing; the carboxyl activator solution can also be added to the polyamino acid aqueous solution and the composite solution respectively to prepare the precursor solution A and the precursor solution B, and when used, the precursor solution A and the precursor solution B are sprayed for mixing.

[0020] In some embodiments, the carboxyl activator solution is added to the polyamino acid aqueous solution and the composite solution to prepare precursor solution A and precursor solution B, and when used, the precursor solution A and the precursor solution B are sprayed and mixed. Specifically, the precursor solution A and the precursor solution B are sprayed through a double channel to form a hydrogel.

[0021] In a second aspect, an in-situ spray polyamino acid hydrogel is obtained by the above preparation method.

[0022] A third aspect provides an application of the above-mentioned in-situ spray polyamino acid hydrogel, wherein the application is used in the preparation of medicines, medical devices or daily chemicals.

[0023] Specifically, the medicine, medical device or daily chemical product has the functions of scalp care and / or hair follicle nourishment.

[0024] Specifically, the daily chemicals include toiletries, cosmetics, and the like.

[0025] The beneficial effects of the present invention are:

[0026] The present invention uses biodegradable and absorbable biomacromolecule polyamino acid materials polyglutamic acid and polyaspartic acid as molecular skeletons, and then under the action of a carboxyl activator, arginine and lysine are used as cross-linking agents, and through an amidation reaction between carboxyl and amino / guanidine groups, diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oils, and plant extracellular vesicles are encapsulated in the interior of a hydrogel 3D cross-linked network structure with viscoelastic behavior to achieve a strong sustained release performance. The three-dimensional network space structure with high water content constructed by the present invention can simulate the natural skin extracellular matrix from the perspective of structure and composition and matrix mechanics (viscoelasticity), reshape the cell physiological microenvironment of damaged scalp tissue, provide a suitable environment for hair follicle cells, promote cell growth and differentiation, interfere with fibroblasts to produce mature collagen and prevent mature collagen deposition, avoid hardening and contraction of surrounding hair follicles, accelerate blood circulation, and promote the proliferation of hair papilla cells; improve the ability of hair papilla cells to migrate, promote hair papilla cells to secrete type III collagen, repair damaged scalp, and have the effect of nourishing hair follicles. In addition, the plant essential oil in the hydrogel composition can act as a penetration enhancer to disturb the lipid structure between keratinocytes in the stratum corneum and the interaction with the intercellular domain of proteins to improve the transdermal absorption rate of the effective ingredient polyamino acid hydrogel in the composition. Plant essential oils also have the advantages of being natural, mild, non-toxic, non-irritating and having excellent cell compatibility, and can be widely used in regenerative medicine, tissue engineering and other fields. DETAILED DESCRIPTION

[0027] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0028] Example 1

[0029] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0030] (1) Dissolve 1 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES (2-morpholineethanesulfonic acid) buffer and stir for later use.

[0031] (2) Dissolve 0.05 g of arginine and 0.3 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, add them dropwise into the amino acid composite solution under continuous stirring to fully disperse and mix them, then add 0.001 g, 0.005 g, and 0.0005 g of Codonopsis pilosula essential oil (Shanghai Yuanye Biotechnology Co., Ltd., purity ≥98%), angelica essential oil (Shaanxi Longzhou Biotechnology Co., Ltd., purity ≥99%), and ginseng extracellular vesicles (for the preparation method of extracellular vesicles, refer to Advanced Science, 2024, 11(17): 2308235, the same below) respectively, and mix them evenly under continuous stirring.

[0032] (3) Adding a carboxyl activator (1 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0033] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0034] Example 2

[0035] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0036] (1) Dissolve 1.2 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0037] (2) Dissolve 0.02 g of arginine and 0.4 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, add them dropwise into the amino acid composite solution under continuous stirring to fully disperse and mix them, then add 0.005 g of Codonopsis pilosula essential oil and 0.0008 g of ginseng extracellular vesicles respectively, and mix them evenly under continuous stirring.

[0038] (3) Adding a carboxyl activator (1.2 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0039] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0040] Example 3

[0041] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0042] (1) Dissolve 1.2 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0043] (2) Dissolve 0.02 g of arginine and 0.1 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, add them dropwise into the amino acid composite solution under continuous stirring to fully disperse and mix them, then add 0.005 g of Codonopsis pilosula essential oil and 0.0008 g of Platycladus orientalis extracellular vesicles in turn and mix them evenly under continuous stirring.

[0044] (3) Adding a carboxyl activator (1.2 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0045] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0046] Example 4

[0047] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0048] (1) Dissolve 1.2 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0049] (2) Dissolve 0.02 g of arginine and 0.1 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, and then add them dropwise to the amino acid composite solution under continuous stirring to fully disperse them for use.

[0050] (3) Adding a carboxyl activator (1.2 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0051] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0052] Example 5

[0053] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0054] (1) Dissolve 1.2 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0055] (2) Dissolve 0.02 g of arginine and 0.1 g of lysine in 8 mL of deionized water to form an amino acid complex solution for later use.

[0056] (3) Adding a carboxyl activator (1.2 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0057] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0058] Example 6

[0059] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0060] (1) Dissolve 1 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0061] (2) Dissolve 0.05 g of arginine in 8 mL of deionized water to form an amino acid solution; then, respectively, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, and then add them dropwise into the amino acid composite solution under continuous stirring to fully disperse and mix them, and then add 0.001 g, 0.005 g, and 0.0005 g of Codonopsis pilosula essential oil, 0.005 g, and 0.0005 g of ginseng extracellular vesicles, respectively, in turn, and mix them evenly under continuous stirring.

[0062] (3) Adding a carboxyl activator (1 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0063] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0064] Example 7

[0065] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0066] (1) Dissolve 1 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0067] (2) Dissolve 0.3 g of lysine in 8 mL of deionized water to form an amino acid solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under continuous stirring, add them dropwise into the amino acid composite solution under continuous stirring to fully disperse and mix them, then add 0.001 g, 0.005 g, and 0.0005 g of Codonopsis pilosula essential oil, 0.005 g, and 0.0005 g of ginseng extracellular vesicles, respectively, in turn, and mix them evenly under continuous stirring.

[0068] (3) Adding a carboxyl activator (1 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0069] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0070] Example 8

[0071] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0072] (1) Dissolve 1 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid Tris (tris(hydroxymethyl)aminomethane) buffer and stir for later use.

[0073] (2) Dissolve 0.05 g of arginine and 0.3 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under constant stirring, add them dropwise into the amino acid composite solution under constant stirring to fully disperse and mix them, then add 0.001 g, 0.005 g, and 0.0005 g of Codonopsis pilosula essential oil, 0.005 g, and 0.0005 g of ginseng extracellular vesicles respectively, and mix them evenly under constant stirring.

[0074] (3) Adding a carboxyl activator (1 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0075] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0076] Example 9

[0077] An in-situ spray polyamino acid hydrogel, the preparation method of which comprises the following steps:

[0078] (1) Dissolve 1 g of polyglutamic acid powder in 10 mL of 1 wt% polyaspartic acid MES buffer and stir for later use.

[0079] (2) Dissolve 0.05 g of arginine and 0.3 g of lysine in 8 mL of deionized water to form an amino acid composite solution; then, dissolve 0.001 g of diaminopyrimidine oxide and 0.001 g of pyrrolidine diaminopyrimidine oxide in 2 mL of ethanol / glycerol solution (the volume ratio of ethanol to glycerol is 1:4) under constant stirring, add them dropwise into the amino acid composite solution under constant stirring to fully disperse and mix them, then add 0.001 g, 0.005 g, and 0.0005 g of Codonopsis pilosula essential oil, 0.005 g, and 0.0005 g of ginseng extracellular vesicles respectively, and mix them evenly under constant stirring.

[0080] (3) Adding a carboxyl activator (1 g each of EDC and NHS) to the composite solutions obtained in step (1) and step (2), respectively, stirring evenly to fully dissolve them, and obtaining an in-situ spray polyamino acid hydrogel precursor solution A and an in-situ spray polyamino acid hydrogel precursor solution B, respectively.

[0081] (4) The in-situ spray polyamino acid hydrogel precursor solution A and the in-situ spray polyamino acid hydrogel precursor solution B described in step (3) are placed in double-channel spray bottles respectively, and then the solution A and the solution B are squeezed out in equal volumes by pressing and fully mixed to obtain the in-situ spray polyamino acid hydrogel.

[0082] Example 10

[0083] The rheological characteristics (stress scanning, frequency scanning), antioxidant properties (DPPH removal, H2O2 removal), cell survival rate (using hair follicle papilla cells to evaluate hydrogel toxicity), hemolysis rate and other properties of the in situ sprayed polyamino acid hydrogel described in Examples 1-9 were tested, and the test results are shown in Table 1.

[0084] Table 1 Physicochemical properties and biological characteristics of in situ sprayable polyamino acid hydrogels

[0085]

[0086] The rheological characteristics of the in situ sprayed polyamino acid hydrogel prepared in Examples 1-9 can be shown by characterization of stress scanning and frequency scanning. Table 1 shows that the mechanical properties of the in situ sprayed polyamino acid hydrogel prepared in Examples 1-9 have good mechanical properties, and its storage modulus is about 10 times that of the loss modulus, and has a certain viscoelastic behavior, which can better simulate the mechanical strength of natural soft tissue, provide a more matching dynamic cell microenvironment for damaged hair follicle cells, accelerate hair follicle cell renewal, promote hair growth and improve hair shedding behavior.

[0087] Good antioxidant properties can improve the disordered inflammatory environment and oxidative stress behavior in the scalp, rebuild the physiological microenvironment of damaged hair follicle cells, maintain scalp health, promote hair growth, and improve hair loss. Table 1 shows that the in-situ spray polyamino acid hydrogel prepared in Examples 1-9 has good antioxidant properties.

[0088] The toxicity of the hydrogel was evaluated using hair follicle papilla cells. The results in Table 1 confirmed that the in situ spray polyamino acid hydrogel prepared in Examples 1-9 had good cell compatibility and could promote cell proliferation, help promote the metabolism of damaged hair follicle cells, accelerate hair follicle cell renewal, promote hair regeneration and reduce hair loss behavior.

[0089] Meanwhile, the hemolysis rate results in Table 1 indicate that the in-situ spray polyamino acid hydrogels prepared in Examples 1-9 have good blood safety.

[0090] In summary, the present invention uses multiple cross-linking technology to cross-link polyamino acid materials, and performs all-round and multi-level biomimetic from the perspective of the composition, function, structure and matrix mechanics (viscoelasticity) of the extracellular matrix of hair follicle cells to provide a stable physical and biological environment for natural hair follicle cells and promote cell proliferation. And by encapsulating diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oil, and plant extracellular vesicles inside the hydrogel 3D cross-linked network structure, the problems of poor hydrophilicity and low bioavailability of active ingredients are avoided, and it is expected to achieve long-term sustained release performance.

[0091] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for preparing an in-situ spray polyamino acid hydrogel, characterized in that: The steps include: dissolving polyglutamic acid and polyaspartic acid to prepare a polyamino acid aqueous solution; Arginine, lysine, diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oil, and plant extracellular vesicles are dissolved and dispersed uniformly to prepare a composite solution; providing a carboxyl activator solution; When in use, the polyamino acid aqueous solution, the composite solution and the carboxyl activator solution are sprayed and mixed.

2. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: The mass concentration of polyglutamic acid in the polyamino acid aqueous solution is 2-20wt%, and the mass ratio of polyglutamic acid to polyaspartic acid is (5-30):

1.

3. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: The solvent for dissolving polyglutamic acid and polyaspartic acid is at least one of MES buffer, Tris buffer and ultrapure water.

4. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: Dissolving arginine and lysine to form an amino acid composite solution; dissolving diaminopyrimidine oxide and pyrrolidine diaminopyrimidine oxide, adding the amino acid composite solution to dissolve, and then adding natural plant essential oil and plant extracellular vesicles to mix evenly; Preferably, the amino acid composite solution is added in a non-all-at-once manner; Preferably, the solvent for dissolving diaminopyrimidine oxide and pyrrolidine diaminopyrimidine oxide is an alcohol solvent, preferably a mixture of ethanol and glycerol.

5. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: The mass concentration of arginine in the composite solution is 1-10wt%, the mass ratio of arginine to lysine is 1:(4-20), and the mass ratio of arginine to diaminopyrimidine oxide, pyrrolidine diaminopyrimidine oxide, natural plant essential oil, and plant extracellular vesicles is 1:(0.01-0.1):(0.01-0.1):(0.05-0.5):(0.001-0.5).

6. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: The natural plant in the natural plant essential oil is at least one of ginger root, codonopsis pilosula, mint, angelica sinensis and millettia reticulata; Or, the plant in the plant extracellular vesicles is at least one of ginseng, aloe vera, Polygonum multiflorum, and Platycladus orientalis leaves.

7. The method for preparing the in-situ spray polyamino acid hydrogel according to claim 1, characterized in that: The carboxyl activator solution is added to the polyamino acid aqueous solution and the composite solution to prepare precursor solution A and precursor solution B. When used, the precursor solution A and the precursor solution B are sprayed and mixed; preferably, the precursor solution A and the precursor solution B are sprayed through a double channel to form a hydrogel.

8. An in-situ spray polyamino acid hydrogel, characterized in that: Obtained by the preparation method described in any one of claims 1 to 7.

9. An application of the in-situ spray polyamino acid hydrogel according to claim 8, characterized in that: The application is application in the preparation of medicines, medical devices or daily chemicals.

10. The use according to claim 9, characterized in that: Medicines, medical devices or daily chemicals have the function of scalp care and / or hair follicle nourishment; Or, the daily chemicals are toiletries or cosmetics.

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