Novel skin repairing preparation

Abstract

Use of human serum albumin in the manufacture of a medicament for improving the skin condition of a subject. The skin repairing preparation comprises human serum albumin and an optional medicinal carrier. The young and injury-free recombinant human serum albumin is used for performing mesoderm injection on a subject, and an excellent skin improvement effect is achieved.

Description

A new skin repair preparation

[0001] Cross-reference to related applications

[0002] This application claims priority to Chinese patent application number CN202311205659.6 and filing date September 15, 2023, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of medical cosmetology, and more particularly to the application of human serum albumin in improving the skin condition of a subject. Background Art

[0004] The mesoderm, located between the epidermis and dermis, plays a key role in the skin's absorption of active ingredients from topical beauty products and its storage of collagen and elastin within the skin. Mesotherapy, first proposed by French physician Marc Piest in 1952, utilizes ultra-micro-penetration technology to deliver high-concentration skin nutrients directly through the epidermis into the subcutaneous tissue in a targeted, layered, and quantitative manner, thereby improving skin condition and combating aging. Currently, this method of delivering nutrients directly to the deeper layers of the skin through specialized injections has become a key anti-aging approach.

[0005] Depending on the different mechanisms of action of different nutrients on the skin, existing mesotherapy has different focuses, mainly including locking in moisture and moisturizing, hemostasis and coagulation, repair and anti-inflammation, and promoting tissue regeneration. Commonly used nutrients include hyaluronic acid, platelet-rich plasma, polydeoxynucleotides, glutathione, vitamin C, etc.

[0006] Human albumin is synthesized in the liver and transported to various tissues and organs via the circulatory system. Intravascular albumin accounts for 30%-40% of total body storage, while 60% of extravascular albumin is distributed in tissues such as the skin, muscle, and intestinal mucosa. Albumin is highly hydrophilic and can freely pass through capillary walls, dynamically exchanging between circulating blood and extracellular fluid, maintaining the balance of colloidal osmotic pressure between the two. Furthermore, albumin has the functions of scavenging free radicals, regulating cell apoptosis, maintaining acid-base balance, and acting as an anti-coagulator. It is a crucial component for maintaining hemodynamic stability and transporting oxygen.

[0007] Summary of the Invention

[0008] The present invention provides use of human serum albumin in the manufacture of a medicament for improving the skin condition of a subject.

[0009] The present invention also provides a skin repair preparation comprising human serum albumin and an optional pharmaceutically acceptable carrier.

[0010] The human serum albumin in the present invention can be human serum albumin prepared from human blood or recombinantly produced. A particularly preferred human serum albumin is young and undamaged human serum albumin.

[0011] In some embodiments, the drug may further contain sodium hyaluronate to improve delivery efficiency and usage experience.

[0012] In some embodiments, the drug may further comprise additional functional components, such as nicotinamide, α-lipoic acid, carnosine, resveratrol, and the like.

[0013] Preferably, the medicament is for mesodermal injection into a subject.

[0014] Those skilled in the art will appreciate that the drug provided by the present invention can be used for patients suffering from skin diseases, and can also be used for healthy subjects who need to improve their skin condition. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Figure 1. Before-and-after comparisons. Photographs of the front and right side of subject 001 before and after treatment show overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 25% to 33%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0016] Figure 2. Before-and-after comparisons. Photographs of the front and right side of subject 002 before and after treatment demonstrate overall skin condition and improvement in brown spots. Under Visia, the brown spot percentile improved from 27% to 92%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0017] Figure 3. Before-and-after comparisons. Photographs of the front and right side of subject 003 before and after treatment show overall skin condition and improvement in brown spots. Visio showed no significant improvement in the brown spot percentile score. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0018] Figure 4. Before-and-after comparisons. Photographs of the front and right side of subject 004 before and after treatment show overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 54% to 69%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0019] Figure 5. Before-and-after comparisons. Photographs of the front and right side of subject 005 before and after treatment demonstrate overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 16% to 19%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0020] Figure 6. Before-and-after comparisons. Photographs of the front and right side of subject 006 before and after treatment show overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 44% to 48%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0021] Figure 7. Before-and-after comparisons. Photographs of the front and right side of subject 007 before and after treatment demonstrate overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 29% to 30%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0022] Figure 8. Before-and-after comparisons. Photographs of the front and right side of subject 008 before and after treatment show overall skin condition and improvement in brown spots. Visio showed no significant improvement in the brown spot percentile score. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0023] Figure 9. Before-and-after comparisons. Photographs of the front and right side of subject 009 before and after treatment demonstrate overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 3% to 11%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0024] Figure 10. Before-and-after comparisons. Photographs of the front and right side of subject 010 before and after treatment show overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 22% to 33%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0025] Figure 11. Before-and-after comparisons. Photographs of the front and right side of subject 011 before and after treatment show overall skin condition and improvement in brown spots. Under Visia, the brown spot percentage improved from 39% to 50%. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0026] Figure 12. Before-and-after comparisons. Photographs of the front and right side of subject 012 before and after treatment show overall skin condition and improvement in brown spots. Vision analysis revealed no significant improvement in the brown spot percentile score. A. Front view, left side before treatment, right side after treatment. B. Right side view, left side before treatment, right side after treatment.

[0027] Figure 13: Viscosity curves of sodium hyaluronate with different molecular weights.

[0028] Figure 14: Viscosity curves of composite preparations of sodium hyaluronate with different molecular weights and recombinant albumin with different concentrations.

[0029] Figure 15: Affinity detection chart of recombinant human serum albumin and various ligands.

[0030] Figure 16: Before and after treatment comparison. Subject 013 was photographed on the right side and front of the face before and after treatment, observing the overall skin condition and improvement in pores and red areas. Under Visia, pores and red areas were effectively improved.

[0031] Figure 17: Before and after treatment comparison. Photographs of the right side of subject 029 before and after treatment show the overall skin condition and improvement in red areas. Under Visia, the red areas were significantly improved.

[0032] Figure 18: Before-and-after comparison. Subject 018's right side was photographed before and after treatment, showing the overall skin condition and improvement in redness, brown spots, and texture. Under Visia, redness, brown spots, and texture were significantly improved. DETAILED DESCRIPTION

[0033] The term "human serum albumin" (HSA) used herein has a well-known meaning in the art, with a CAS NO of 70024-90-7 and a molecular weight of 66 kD.

[0034] The term "human serum albumin" or "HSA" includes wild-type human serum albumin (see, for example, Gene name: ALB; NCBI ID: 213; UniProtKB-P02768), as well as functional mutants or modifications thereof. Those skilled in the art will appreciate that wild-type human serum albumin can be modified to a certain extent, such as by the addition, deletion, or substitution of one or more amino acid residues, without substantially affecting its biological function. Therefore, the term "human serum albumin" as used herein also encompasses such modified human serum albumin mutants or modifications.

[0035] Human serum albumin can be prepared by a variety of methods. Commonly available commercially available HSA is often derived from human blood, but it can also be produced through recombinant methods. HSA products prepared by different processes exhibit varying performance, including in terms of degree of oxidation, glycation, and biological activity.

[0036] In this document, "human serum albumin" may also be referred to as "human albumin", "human albumin", etc.

[0037] Young healthy human serum albumin

[0038] As used herein, the term "young, undamaged human serum albumin" or "young, undamaged HSA" refers to HSA in a fresh state without significant damage. "Damage" herein includes oxidation, glycation, carbonylation, and the like.

[0039] The skilled artisan will appreciate that the term "young, undamaged" HSA does not strictly require that the HSA be absolutely free of damage. It is very difficult to produce HSA without any damage. For the purposes of the present invention, HSA containing limited damage is still acceptable and can still be referred to as young, undamaged HSA.

[0040] "Young, intact" HSA typically exhibits at least one, for example, two, three, and preferably four, of the following four properties: (1) a higher proportion of free sulfhydryl groups at the Cys-34 residue; (2) lower levels of advanced glycation end products (AGEs); (3) lower levels of carbonylation; and (4) lower levels of homocysteine. Young, intact HSA typically has a higher proportion of free sulfhydryl groups and lower levels of AGEs, carbonyls, and homocysteine.

[0041] In some embodiments, "young, intact" HSA exhibits at least one of the following four properties, such as two, three, and preferably four: (1) the proportion of free sulfhydryl groups at Cys-34 residues as determined by the Ellman method is greater than 50%, such as 70%, in particular 80%, preferably 90%, and more preferably greater than 95%; (2) the level of advanced glycation end products (AGEs) as determined by ELISA (CLOUD-CLONE Co., CEB353Ge) is less than 60 μg / g protein, preferably 40 μg / g protein, and more preferably less than 30 μg / g protein; (3) the level of carbonyls as determined by the Protein Carbonyl Content Assay Kit is less than 1.7 nmol / mg protein, preferably less than 1.5 nmol / mg protein; (4) the level of homocysteine ​​as determined by ELISA (Jianglai, JL10022) is less than 5 nmol / g protein, preferably 3.5 nmol / g protein, and more preferably less than 2 nmol / g protein.

[0042] In some embodiments, the present invention provides a preparation of young, undamaged human serum albumin (HSA) that exhibits at least one, e.g., two, three, and preferably four, of the following four properties when compared to an endogenous HSA preparation prepared from the plasma of a young individual: (1) a higher proportion of free sulfhydryl groups at Cys-34 residues; (2) lower levels of advanced glycation end products (AGEs); (3) lower levels of carbonylation; and (4) lower levels of homocysteine.

[0043] The term "young individual" generally refers to a person under 30 years old, preferably under 18 years old, more preferably under 3 years old, or even under 1 year old.

[0044] The term "preparation" refers to a material prepared by a process. For example, an HSA preparation can be an HSA sample prepared from a specific individual or from multiple individuals, comprising a population of human serum albumin molecules. A "preparation" can be a packaged commercial product or a crude product prepared in the form of a solution or lyophilized powder.

[0045] Free sulfhydryl group: Wild-type HSA has only one free sulfhydryl group, located at residue Cys-34. Those skilled in the art will readily appreciate that functional HSA mutants may contain slightly different amino acid sequences than wild-type HSA, and thus the Cys-34 residue may be located at a slightly different position than position 34. This can be readily determined using methods such as homology alignment.

[0046] In some embodiments, the proportion of free sulfhydryl groups on Cys-34 residues is determined by the Ellman method. Preferably, the proportion of free sulfhydryl groups is greater than 70%, particularly 80%, preferably 90%, and more preferably greater than 95%.

[0047] AGE: Serum albumin is a plasma protein that is highly sensitive to glycation. This process, also known as the Maillard reaction, is a slow, non-enzymatic reaction that initially involves the connection of glucose or a derivative to the free amine groups of albumin to form a reversible Schiff base product, resulting in the formation of a stable fructosamine residue (ketoamine) after the Amadori rearrangement. The Amadori product can then cyclize to form a pyranose or furanose adduct. Further modifications of these early glycation products, such as rearrangement, oxidation, polymerization, and cleavage, produce irreversible conjugates called advanced glycation end products (AGEs). The mass concentration of AGEs was measured using an enzyme-linked immunosorbent assay (ELISA) kit according to the manufacturer's instructions (CLOUD-CLONE Co., CEB353Ge). The results were normalized by the HSA concentration of each reaction and expressed as micrograms of AGE per gram of protein.

[0048] In some embodiments, the AGE level is determined by ELISA (CLOUD-CLONE Co., CEB353Ge).Preferably, the AGE level is less than 60 μg / g protein, preferably less than 40 μg / g protein, and more preferably less than 30 μg / g protein.

[0049] Carbonyl: Carbonylation here refers to the formation of carbonyl groups on amino acid residues of HSA. The Protein Carbonyl Content Assay Kit (ab126287) is used to quantify carbonyl groups in HSA. This method is based on the reaction of DNPH with protein carbonyls, which forms DNP hydrazones and can be quantified using a microplate reader at an absorbance of 375 nm. According to the manual, the molar concentration of carbonyl groups was quantified using the Protein Carbonyl Content Assay Kit (Abcam, ab126287). The results were normalized by the HSA concentration of each reaction and expressed as nanomoles of carbonyl per milligram of protein.

[0050] In some embodiments, the level of carbonyl is determined by a Protein Carbonyl Content Assay Kit. Preferably, the level of carbonyl is less than 1.7 nmol / mg protein, preferably less than 1.5 nmol / mg protein.

[0051] Homocysteine ​​(HCY): Homocysteine ​​(HCY) is metabolized from the essential amino acid Met and is found to be involved in post-translational modification (PTM) of proteins. HCY can be linked to proteins via an isopeptide bond (N-Hcy-protein) with a lysine (Lys) residue or via a disulfide bond (S-Hcy-protein) with a Cys-34 residue. N-homocystineization is an important post-translational modification that affects the structure and function of proteins and causes protein damage. N-homocystineization can be measured by Hcy-thiolactone produced only by Hcy. The molar concentration of Hcy can be quantitatively determined by ELISA and HPLC. In our study, the Hcy molar concentration was measured by ELISA according to the manufacturer's instructions (Jianglai, JL10022). The results were normalized by the HSA concentration of each reaction and expressed as nmol Hcy per gram of protein.

[0052] In some embodiments, the level of homocysteine ​​is determined by ELISA (Jianglai, JL10022). Preferably, the level of homocysteine ​​is less than 5 nmol / g protein, preferably less than 3.5 nmol / g protein, more preferably less than 2 nmol / g protein.

[0053] Those skilled in the art will appreciate that the above four parameters can be measured by other generally accepted methods, and that the numerical results of the parameters measured by different methods may be different. Those skilled in the art can easily compare the results measured from different methods through limited experiments.

[0054] In a preferred embodiment, the proportion of free sulfhydryl groups on the Cys-34 residue is greater than 80%, the AGE level is less than 30 μg / g protein, the carbonyl level is less than 1.5 nmol / mg protein, and the homocysteine ​​level is less than 2 nmol / g protein.

[0055] Young, undamaged HSA preparations can be recombinantly produced or purified from human plasma. Preferably, the HSA preparation is recombinantly produced.

[0056] The rHSA used in this study was from Progen and was produced by recombinant means.

[0057] Skin repair preparations

[0058] Another aspect of the present invention provides a skin repair preparation comprising: human serum albumin, and optionally a pharmaceutically acceptable carrier.

[0059] As used herein, the term "pharmaceutically acceptable carrier" refers to a solid or liquid diluent, filler, antioxidant, stabilizer, or other substance that can be safely administered. Depending on the route of administration, various carriers well known in the art can be used, including, but not limited to, sugars, starch, cellulose and its derivatives, maltose, gelatin, talc, calcium sulfate, vegetable oils, synthetic oils, polyols, alginic acid, phosphate buffer, emulsifiers, isotonic saline, and / or pyrogen-free water.

[0060] The term "optional" means that it may or may not be present. For example, "optional pharmaceutically acceptable carrier" means that the preparation may or may not contain a pharmaceutically acceptable carrier, which can be determined by those skilled in the art based on the specific circumstances.

[0061] As used herein, the term "therapeutically effective amount" refers to an amount of an active substance sufficient to elicit a biological or medical response in a subject as desired by a clinician. A "therapeutically effective amount" can be determined by one skilled in the art based on factors such as the route of administration, the subject's weight, age, and condition. For example, a typical daily dose may range from 0.01 mg to 100 mg of active ingredient per kg body weight.

[0062] The drug provided by the present invention can be prepared into a clinically acceptable dosage form such as an injection. The drug of the present invention can be administered to a subject by any appropriate route, for example, by mesodermal injection or the like.

[0063] Medical and aesthetic applications of human serum albumin

[0064] Based on an understanding of the fundamental physicochemical properties and functions of human serum albumin, the present invention utilizes human serum albumin to promote skin regeneration and repair, as well as anti-aging. In some embodiments, mesodermal injections are used as a treatment to observe the effectiveness of albumin in improving the overall condition of the skin and alleviating skin concerns such as spots, wrinkles, fine lines, pores, brown spots, and red areas (inflammation).

[0065] Those skilled in the art will appreciate that the human serum albumin in the present invention may be human serum albumin obtained from human blood or recombinantly produced.

[0066] A particularly preferred human serum albumin is young and undamaged human serum albumin. The molecular structure of the young and undamaged recombinant human serum albumin prepared using genetic engineering technology is completely consistent with the naturally synthesized albumin of the human body. In addition to having the basic physiological functions of albumin, since the natural conformation and physicochemical properties of the protein are maintained to the greatest extent during the preparation process, it has strong reducing power, low glycation modification, low carbonylation and young and undamaged characteristics of homocysteine ​​modification (see PCT international patent application PCT / CN2021 / 112093), thus having stronger hydrophilicity, antioxidant, anti-glycation, anti-inflammatory and other functions. The above characteristics are all basic elements required for ideal skin anti-aging and repair products. Therefore, young and undamaged recombinant human serum albumin is expected to become a new star in mesotherapy in addition to traditional commonly used nutrients.

[0067] The young, undamaged recombinant human serum albumin used in the embodiments of the present invention is produced by Progi Corporation and is obtained by genetic engineering means using Pichia yeast fermentation and purification. The protein conformation is consistent with natural human serum albumin, with a purity exceeding 99.999999%. The proportion of Cys34 amino acid residues that provide reducing power in the reduced state exceeds 99%, and the overall protein post-modification levels such as glycation, carbonylation and homocysteine ​​are lower than those of commercially available blood-derived human serum albumin (see PCT / CN2021 / 112093).

[0068] In some embodiments of the present invention, 12 volunteer subjects completed three consecutive treatments of young, non-damaging recombinant human serum albumin mesoderm injections, and the treatments were evaluated using the VISIA system before and one month after each treatment. To minimize factors that could interfere with efficacy evaluation, subjects did not receive any other medical aesthetic treatments other than daily skincare from one month before treatment to one month after the last treatment. Furthermore, fixed equipment, a fixed environment, and relatively fixed operators were used to minimize potential interference and systematic errors during the injection, photography, and data evaluation processes.

[0069] In other embodiments of the present invention, after 4 months of follow-up observation in 30 volunteer subjects, it was proved that the use of recombinant albumin dermal mesotherapy injection for mesodermal injection can effectively improve the overall condition of the skin, especially pigmentation, pores, texture and sensitivity problems, and achieved statistically significant improvement.

[0070] Young, undamaged recombinant human serum albumin is safe and well tolerated for mesodermal injection, and the therapeutic effect analysis uses a combination of the GAIS method and the VISIA system score. The GAIS method is used by two professional doctors to score the overall skin improvement based on photographs and calculate the average value. The VISIA system score is a percentile quantitative value derived from the system's own data. It was found that the vast majority of subjects achieved varying degrees of improvement in overall skin condition, spots, wrinkles, texture, pores, brown spots, and red areas (inflammation). The VISIA system can also derive absolute value scores, which can be verified with the results of the GAIS method and VISIA percentile scores.

[0071] Since the viscosity of the young, non-damaged recombinant albumin solution is low, slight leakage of the syringe is prone to occur during mesodermal injection, as well as leakage caused by skin tissue extrusion when the pinhole is closed, which reduces the drug delivery efficiency. At the same time, the young, non-damaged recombinant albumin solution is very easy to be absorbed and metabolized through the blood-interstitial fluid substance exchange in the dermis, and the effect is maintained for about 1 week, and it is impossible to continue to stay in the dermis to play a role. Therefore, the present invention further explores the composite formula of young, non-damaged recombinant albumin and sodium hyaluronate, and adjusts the overall viscosity of the composite formulation by adjusting the ratio of high, medium and low molecular weight sodium hyaluronate, thereby optimizing the product delivery efficiency and bioavailability, so that the effect is maintained for more than 6 weeks, and the use experience of doctors and patients is also improved.

[0072] Since sodium hyaluronate in different molecular weight ranges has different viscosities, filling and supporting capacities, and hydrating and moisturizing properties, the present invention attempted to mix recombinant human serum albumin with high molecular weight (2050kD-2950kD), medium molecular weight (800kD-1370kD), and low molecular weight (330kD-363kD) sodium hyaluronate in different proportions. The viscosity and stability of the composite formulations at different compounding ratios were tested, and the results are shown in Table 5 and Figure 13.

[0073] The present invention also attempted to adjust the albumin concentration in the process of adjusting the composite formulation of albumin and sodium hyaluronate, and found that the albumin concentration had little effect on the overall viscosity and stability of the composite formulation. The specific data are shown in Table 6, Table 7 and Figure 14.

[0074] Taking advantage of the fact that albumin can bind to other biomacromolecules such as proteins and lipids, as well as various small molecule compounds, and can provide a slow and sustained release as a natural drug carrier, the present invention further attempted to add nicotinamide, α-lipoic acid, carnosine, resveratrol, and other components with skin repair and anti-aging functions to young, undamaged recombinant human serum albumin to form a composite formula. The affinity of the above components to young, undamaged human serum albumin was preliminarily observed, and the results are shown in Table 8 and Figure 15.

[0075] Example

[0076] Example 1: Mesodermal Therapy Trial Using Young Undamaged Recombinant Human Serum Albumin

[0077] In this study, 12 female subjects (aged 27-41) received three monthly injections of young, non-damaging human serum albumin mesoderm. To avoid interference with efficacy evaluation, all subjects did not receive any other cosmetic procedures from one month before the first treatment until one month after the last treatment.

[0078] The subjects collected data before the first treatment, one month after the first treatment, one month after the second treatment, and one month after the third treatment. The data before the first treatment was used as the initial baseline value for comparison and evaluation of the subsequent treatment effect. The treatment effect was evaluated by taking photos using a VISIA skin analyzer (produced by Canfield, USA, model 7.1.0). The image data of the front, left (45°) and right (45°) positions were collected under standard white light, 365nm ultraviolet light and cross-polarized light (Figures 1-12). Two professional doctors were asked to give GAIS scores for the overall improvement of the facial skin based on the image data, and then the mean was calculated (Table 1). In addition, the system's own data export and analysis functions were used for qualitative and quantitative evaluation (Tables 2-4).

[0079] The specific operation steps are as follows: the subject first cleans the face, takes a photo with the VISIA system after the skin is naturally dry, and then applies anesthetic ointment (Tsinghua Unisplendour) for 30 minutes. After removing the anesthetic ointment, a young and non-damaged recombinant human serum albumin (see PCT / CN2021 / 112093) mesodermal injection is performed. The albumin concentration is 200mg / mL. The injection instrument uses a DermaShine electronic syringe, Panace-DS-30 model, and the injection needle uses a DermaShine brand disposable sterile injection needle. The injection depth is 0.8mm-1.2mm, the single injection volume is 0.0167ml-0.0250ml, and a total of 5ml is injected. After the full face injection is completed, apply a medical moisturizing dressing to calm the skin for 20 minutes, and then follow the precautions after routine mesodermal treatment for care.

[0080] Example 2: Young, non-damaged recombinant human serum albumin mesodermal therapy can improve the facial skin condition of subjects

[0081] The fourth evaluation results collected in the experiment described in Example 1 (ie, the VISIA photography results one month after the third treatment) were directly compared with the initial baseline status collected before the first treatment, and the results shown in Figures 1 to 12 were obtained.

[0082] The overall skin condition of most subjects was improved to varying degrees. The improvements mainly included skin translucency, reduced acne marks and pigmentation, refined skin texture, smaller pores, improved acne inflammation, reduced redness, and reduced fine lines under the eyes.

[0083] Two professional physicians evaluated the overall improvement before and after treatment using the GAIS scoring method based on VISIA system data, and the average score was calculated. The scores are shown in Table 1, where 0 indicates no improvement, 1 indicates mild improvement, 2 indicates moderate improvement, and 3 indicates marked improvement. Of the 12 subjects, 2 (05 and 07) scored 0 for no improvement. All other 10 subjects achieved varying degrees of overall improvement, with 8 achieving mild, mild-to-moderate, or moderate improvement.

[0084] Table 1. GAIS overall improvement score before and after treatment

[0085] Example 3: Young, non-damaged recombinant human serum albumin mesotherapy can alleviate facial skin problems in subjects

[0086] The fourth evaluation results collected in the experiment described in Example 1 (i.e., the VISIA image results one month after the third treatment) were directly compared with the initial baseline state collected before the first treatment. The image information was quantified and exported using the VISIA system's built-in software. The quantitative results shown in Tables 2-4 were obtained for spots, wrinkles, texture, pores, brown spots, and red areas (inflammation).

[0087] The evaluation in this embodiment is expressed in percentile form, that is, the percentile of the status of each VISIA photo record within the system comparable range is given. The larger the value, the greater the probability of exceeding the comparable object, that is, the better the skin condition.

[0088] Table 2 shows the scores of various indicators before and after treatment derived from the analysis of VISIA images taken from the front side (front view), among which 41.7% of the subjects had improved spots, 41.7% of the subjects had improved wrinkles, 41.7% of the subjects had improved texture, 41.7% of the subjects had improved pores, 75% of the subjects had improved brown spots, and 25% of the subjects had improved red areas (inflammation).

[0089] Table 2. Percentile score table of frontal image information before and after treatment

[0090] Table 3 shows the scores of various indicators before and after treatment derived from the analysis of the VISIA images taken on the right. Among them, 50% of the subjects had improved spots, 41.7% of the subjects had improved wrinkles, 41.7% of the subjects had improved texture, 41.7% of the subjects had improved pores, 75% of the subjects had improved brown spots, and 25% of the subjects had improved red areas (inflammation).

[0091] Table 3. Percentile score table of right image information before and after treatment

[0092] Table 4 shows the scores of various indicators before and after treatment derived from the analysis of the VISIA images taken on the left. Among them, 50% of the subjects had improved spots, 33.3% of the subjects had improved wrinkles, 8.3% of the subjects had improved texture, 33.3% of the subjects had improved pores, 66.7% of the subjects had improved brown spots, and 50% of the subjects had improved red areas (inflammation).

[0093] Table 4. Percentile score table of left image information before and after treatment

[0094] Example 4: Exploration of Sodium Hyaluronate Concentration and Ratio

[0095] Young, undamaged recombinant human serum albumin has low viscosity, and some protein leaks out after mesoderm injection, impacting the product's effectiveness and user experience. Therefore, the inclusion of sodium hyaluronate in the product formula is considered. This increases product viscosity and reduces leaks while also leveraging hyaluronate's exceptional moisturizing and water-locking properties. By referring to the viscosity of existing products on the market, we screened sodium hyaluronate types and concentrations, testing the viscosities of low-, medium-, and high-molecular-weight sodium hyaluronate at different concentrations, and plotting viscosity curves.

[0096] The experimental procedure was as follows: Sodium hyaluronate solutions of varying concentrations and molecular weights were prepared, and the viscosity was measured using a viscosity tester. Viscosity curves were plotted using a #3 rotor at 30 rpm. Sodium hyaluronate was purchased from Shandong Zhongshan Biotechnology Co., Ltd., and the viscosity tester, model SNB-1, was purchased from Shanghai Hengping Instrument Factory.

[0097] Table 5: Viscosity analysis of sodium hyaluronate with different molecular weights and concentrations

[0098] Note: - indicates the viscosity exceeds the upper limit of instrument detection.

[0099] Example 5: Exploration of viscosity of composite preparations of recombinant human serum albumin and sodium hyaluronate at different concentrations

[0100] Combined with the viscosity curves of sodium hyaluronate with different molecular weights, different concentrations of sodium hyaluronate and recombinant human serum albumin were selected to prepare mixed solutions, and the viscosity of the mixed solutions was tested.

[0101] The experimental procedure was as follows: Sodium hyaluronate (high, medium, and low molecular weight) was selected and prepared at 0.5% and 1% solutions, respectively. These solutions were then mixed with recombinant human serum albumin to ensure final concentrations of 0, 25, 50, 75, and 100 mg / ml. Viscosity was measured using a viscometer, and viscosity curves were plotted. The test conditions were: rotor #3, 30 rpm. Sodium hyaluronate was purchased from Shandong Zhongshan Biotechnology Co., Ltd., and the viscosity measuring instrument, model SNB-1, was purchased from Shanghai Hengping Instrument Factory.

[0102] Table 6: Viscosity analysis of sodium hyaluronate and recombinant human serum albumin at different concentrations

[0103] Example 6: Determination of compound formulation formula

[0104] Based on the viscosity curve observed in Example 5, and taking full advantage of the respective characteristics of high, medium, and low molecular weight sodium hyaluronate, it is proposed to compound the above-mentioned different molecular weight sodium hyaluronates in different proportions and study the viscosity and stability of the compounded solutions.

[0105] The experimental procedure was as follows: Sodium hyaluronate (high, medium, and low molecular weight) was selected and prepared at concentrations of 0.5% and 1% according to Table 7. The solutions were then mixed with recombinant human serum albumin to ensure final concentrations of 50 and 100 mg / ml, respectively. Viscosity was measured using a viscometer, and viscosity curves were plotted. Testing conditions: rotor #3, 30 rpm. Sodium hyaluronate was purchased from Shandong Zhongshan Biotechnology Co., Ltd., and the viscosity measuring instrument, model SNB-1, was purchased from Shanghai Hengping Instrument Factory.

[0106] After comprehensive comparison of viscosity and stability research data, Scheme 1 was determined as the final formulation scheme, and a preliminary stability study was conducted. The accelerated test at 25°C has been carried out for more than 9 months, and the formulated preparation is still stable.

[0107] Table 7: Analysis of the formula of sodium hyaluronate and recombinant human serum albumin compound preparation

[0108] Example 7: Study on the affinity of functional small molecule compounds with recombinant albumin

[0109] Based on the formulation of Scheme 1 determined in Example 6, we further investigated the binding ability of functional small molecule compounds to recombinant albumin. These small molecule compounds included nicotinamide (NAD), L-carnosine, and α-lipoic acid. The study was conducted using optical surface plasmon resonance (SPR) using a Cytiva / Biacore 8K molecular interaction analyzer.

[0110] Experimental Procedure: A HAS-CM5 chip was prepared by coupling a protein sample to a Series S Sensor Chip CM5 chip. The small molecule ligand was accurately weighed, dissolved in an appropriate solvent, and diluted to a working concentration. The affinity between the receptor and the ligand was measured. The test data was automatically processed by BIAcore™ Insight Evaluation Software according to the single-cycle kinetics / steady state affinity model to calculate the association rate constant ka, the dissociation rate constant kd, and the dissociation constant KD (kd / ka).

[0111] The affinity of recombinant human albumin and ligands was tested respectively. The affinity constants KD were Naproxen: 260μM, Nicotinamide: 127μM, L-carnosine: 165μM and α-lipoic acid greater than 2000μM. The affinity results of positive control Naproxen and negative control Quinine were used as reference and the affinity constant KD range of protein and small molecule binding was (10 -3 -10 -6 ), it can be seen that nicotinamide and L-carnosine can interact specifically with recombinant human albumin with strong affinity; α-lipoic acid has no specific interaction with recombinant human albumin.

[0112] Table 8: Affinity analysis results of recombinant albumin and small molecules

[0113] Example 8: Evaluation of the efficacy and safety of albumin in improving skin quality

[0114] Clinical data

[0115] Volunteer selection criteria:

[0116] (1) The subjects were aged 18 years or older and in good health;

[0117] (2) Clinical manifestations include dull facial complexion, flushing and sensitivity, dry lines and fine lines;

[0118] (3) had not received other facial cosmetic treatments in the 3 months before the trial and during the entire observation period;

[0119] (4) Informed consent for the project and signing of the project informed consent form;

[0120] (5) Willing to comply with the trial requirements and complete the entire trial, and able to complete follow-up as required.

[0121] Volunteer exclusion criteria:

[0122] (1) Pregnant or breastfeeding women;

[0123] (2) Those who have taken oral or topical drugs prohibited by the test (glucocorticoids, immunosuppressants and anti-allergic drugs) within 14 days before the test, or have participated in similar tests or received facial treatment in the past three months;

[0124] (3) Skin diseases (e.g., psoriasis, eczema, skin cancer, etc.), or obvious erythema, wounds, abrasions, tattoos, etc. on or near the test area;

[0125] (4) those with coagulation disorders or those using prescription or over-the-counter blood-thinning drugs (anticoagulants);

[0126] (5) Severe complications (such as heart disease), autoimmune diseases, systemic blood diseases, exacerbation of chronic physical diseases, etc.;

[0127] (6) Immunosuppressive diseases (such as HIV positive) and the use of immunosuppressive drugs;

[0128] (7) History of diseases that are prone to irritation, such as recurrence of herpes in the treatment area;

[0129] (8) Those with severe scar tissue;

[0130] (9) Experts or professionals believe that there are other iatrogenic reasons that may affect the test results;

[0131] (10) Participating in other clinical trial researchers;

[0132] (11) Non-voluntary participants or those who are unable to complete the required content according to the test requirements.

[0133] Experimental materials and methods

[0134] Test materials

[0135] (1) Recombinant albumin dermal mesotherapy injection (manufacturer: Shenzhen Progi Pharmaceutical Technology Co., Ltd.);

[0136] (2) VISIA skin image analyzer (produced by Canfield, USA, and distributed by Guangzhou Hanma);

[0137] (3) Dermalab multifunctional skin tester;

[0138] (4) DermaShine electronic syringe.

[0139] Preparation before treatment

[0140] Before treatment, the patient was informed of the treatment method, postoperative reactions and precautions and signed a preoperative informed consent form.

[0141] After cleansing, five photos of the patient's face were taken in the same environment, using the same camera, at the same angle and brightness, from the front, at 45 degrees to the left and right sides, and at 90 degrees to the left and right sides.

[0142] The VISIA skin detector conducts a full-scale monitoring of the skin's spots, wrinkles, texture, pores, UV spots, brown spots, red areas and porphyrin from the left, frontal and right sides respectively, and then the multifunctional skin detector detects the skin's erythema and melanin index.

[0143] Treatment

[0144] Mesotherapy injections were performed using a recombinant albumin dermal mesotherapy sample at an albumin concentration of 100mg / mL. The injection instrument used was a DermaShine Panace-DS-30 electronic syringe with a DermaShine disposable sterile needle. The injection depth ranged from 0.8mm to 1.2mm, and the single injection volume ranged from 0.0167ml to 0.0250ml, with a total of 5ml injected throughout the face. After the injection, a medical moisturizing dressing was applied to calm the skin for 20 minutes. Subsequent care was followed according to standard post-mesotherapy precautions.

[0145] Once every four weeks, for a total of three times.

[0146] Treatment time points: week 0, week 4, week 8 (W0, W4, W8);

[0147] Testing time points: Week 0, Week 4, Week 8, Week 12 (W0, W4, W8, W12). Participants were revisited at W0 (before the first treatment), W4 (before the second treatment), W8 (before the third treatment), and W12 (4 weeks after treatment) and photos and skin tests were taken with the help of instruments.

[0148] All subjects will return to the research center for efficacy and safety assessments at 4, 8, and 12 weeks after the first treatment.

[0149] Clinical effect evaluation

[0150] At week 0 (W0) before treatment and week 12 (W12) after the first treatment, the investigators and the subjects evaluated:

[0151] (1) Dermlab facial erythema index and melanin index.

[0152] (2) Analysis of VISIA photo data.

[0153] (3) Assessment of facial improvement effect: -1 point = worsening, 0 point = no improvement, 1 point = partial improvement, 2 points = significant improvement, 3 points = very significant improvement.

[0154] (4) Two doctors who were not involved in the treatment compared the clinical data of the subjects to perform GPS scoring on facial fine lines, spots, and capillary dilation. Wrinkles: Mild wrinkles are fine lines that appear only under certain facial expressions, such as the shallow wrinkles around the eyes when smiling. These wrinkles are usually short and shallow, few in number, and more limited in distribution = 1-2 points; moderate wrinkles are shallow wrinkles that can be clearly seen even when not making an expression, with moderate length, and may be distributed on the forehead, corners of the eyes, corners of the mouth, etc. = 3-4 points; severe wrinkles are deep and long wrinkles, with a large number and widely distributed in various areas of the face. They may be accompanied by skin depression and sagging = 5-6 points. Pigmentation: Minor light pigmentation: A small number of lighter spots on the face, such as freckles or mild sunburn, are small in diameter, may range from a few to a dozen, and are relatively dispersed = 1-2 points; Moderate pigmentation: A moderate number of slightly darker, larger spots, which may be concentrated in specific areas, such as the cheeks or bridge of the nose = 3-4 points; Extensive dark pigmentation: A large number of darker spots, including melasma, are large in diameter, numerous, and widely distributed on the face = 5-6 points. Telangiectasia: Minor telangiectasia: A small number of subtle red streaks on the face, difficult to detect without close observation, with a relatively limited distribution = 1 point; Moderate telangiectasia: More pronounced red streaks, relatively numerous, distributed on areas such as the cheeks, making them easily noticeable = 2-3 points; Severe telangiectasia: A large number of pronounced telangiectasias on the face, with noticeable and widespread red streaks, which may affect the evenness of facial skin tone = 4 points.

[0155] Safety Assessment

[0156] During the follow-up visit, the subjects were evaluated for adverse reactions and asked whether the original symptoms of the target area worsened or new skin lesions appeared, such as itching, dryness, desquamation, rash, etc., and whether it interfered with daily life.

[0157] Adverse reactions are graded as follows: Severe - The adverse reaction interferes with daily activities, the subject experiences significant symptoms, is unbearable, and treatment discontinuation is necessary; Moderate - The adverse reaction interferes with daily life, the subject experiences significant symptoms but is tolerable, and treatment discontinuation is not necessary; Mild - The adverse reaction is tolerable and occasionally felt by the subject. If the responsible physician deems the subject unsuitable for continued trial or the subject requests discontinuation, the trial will be discontinued for that subject, and the reason and time of discontinuation will be recorded.

[0158] Statistical analysis

[0159] SPSS software was used for data processing. The measurement data were expressed as mean ± standard deviation (x ± s). The t test was used for statistical analysis. The difference was considered statistically significant when p < 0.05.

[0160] Study Results

[0161] Demographic data

[0162] This study included 30 female subjects aged 27-53 years. They presented with varying degrees of facial symptoms, including dull skin tone, dark spots, redness, sensitivity, dry lines, and fine lines.

[0163] Clinical effect evaluation

[0164] Skin tester evaluation

[0165] The subjects were measured for erythema and melanin index using a Dermlab skin detector. The left or right side was selected by random number table to measure the erythema index (EI), and the remaining side was measured for the melanin index (MI). Five points were measured, including 2 cm above the eyebrow, the outer side of the outer canthus, the highest point of the cheekbone, 2 cm next to the corner of the mouth, and the midpoint of the line connecting the corner of the mouth and the mandible. The five measured values ​​were averaged and a paired sample t-test was performed. The data all obeyed the normal distribution, and the measured data before and after treatment were significantly correlated. Melanin index (Melanin) is an indicator for evaluating skin pigmentation, which is used to evaluate the level of pigmentation on the skin. The higher the index, the more pigment on the skin. Erythema index (Erythema) is an indicator for evaluating the degree of skin redness, which is used to evaluate the level of redness (hemoglobin) on the skin. The higher the index, the higher the degree of redness of the skin. The statistical analysis results are as follows:

[0166] Melanin index paired sample statistics

[0167] P<0.05

[0168] Statistics of paired samples of erythema index

[0169] P<0.05

[0170] The analysis results showed that the melanin index (MI) and erythema index (EI) decreased after treatment compared with the mean values ​​before treatment, and the decrease was statistically significant.

[0171] VISIA Assessment

[0172] After the subjects took their VISIA photos, the photos were analyzed using VISIA software to generate scores for various indicators. The average of the scores for the same volunteer's frontal, left 45°, and right 45° images taken at the same time was used for a paired-sample t-test. The analyzed data conformed to a normal distribution, and the scores before and after treatment showed a significant correlation. The statistical analysis results are as follows:

[0173] Visia texture score paired sample statistics

[0174] P<0.05

[0175] Pore ​​score paired sample statistics

[0176] P<0.05

[0177] Statistics of brown spot paired samples

[0178] P<0.05

[0179] Red zone paired sample statistics

[0180] P<0.05

[0181] From the above statistics, it can be seen that the characteristic scores of texture, pores, brown spots, and red areas in the VISIA scores before and after treatment were all reduced, and the p values ​​were all less than 0.05, indicating that the improvements in the above observation indicators were statistically significant.

[0182] After analysis of pigmentation and wrinkles, although the overall scores were slightly reduced, the p-value was greater than 0.05, and the reduction result was not statistically significant.

[0183] GPS Rating

[0184] A professional physician not involved in the treatment performed GPS scoring on pigmentation, telangiectasia, and wrinkles before and after treatment. Paired sample t-tests were performed on the scores. The scores were found to be normally distributed, and there was a significant correlation between the pre- and post-treatment scores.

[0185] The statistical analysis results are as follows:

[0186] Pigment spot GPS score paired sample statistics

[0187] P<0.05

[0188] Paired sample statistics of telangiectasia GPS score

[0189] P<0.05

[0190] Statistics of paired samples of wrinkle GPS scores

[0191] P<0.05

[0192] Results showed that after treatment, the subjects experienced statistically significant improvements in pigmentation, telangiectasia, and wrinkle scores. Figures 16-18 show representative examples of significant improvements in pores, red areas, brown spots, and texture, respectively.

[0193] Subjective rating

[0194] The subjects scored the facial improvement effect according to the GAIS scale. The scoring results are as follows:

[0195] GAIS score

[0196] Among them, 7 cases were very significantly improved, 12 cases were significantly improved, and 10 cases were partially improved, with an improvement rate of 96.7%.

[0197] Safety Assessment

[0198] During the 4-month study, no serious adverse reactions were reported and the drug was well tolerated by the subjects.

[0199] in conclusion

[0200] This study evaluated the safety and efficacy of recombinant albumin dermal mesotherapy injection in improving problematic skin.

[0201] After four months of follow-up observation, Dermlab's skin erythema-melanin index (EMI) showed a statistically significant decrease after treatment compared to pre-treatment. VISIA photo analysis revealed statistically significant decreases in skin texture, pores, brown spots, and red areas. Professional physicians' objective evaluation of GPS scores revealed statistically significant reductions in pigmentation, telangiectasia, and wrinkle scores. Subjective feedback from participants also confirmed the effectiveness of the treatment, with an improvement rate of 96.7%.

[0202] During the clinical study, no serious adverse reactions were reported and the subjects tolerated the drug well.

[0203] In summary, the use of recombinant albumin dermal mesotherapy for mesodermal injection treatment can effectively improve skin texture, pigmentation and sensitivity problems.

[0204] The implementation methods of the present disclosure are not limited to those described in the above embodiments. Without departing from the spirit and scope of the present disclosure, ordinary technicians in this field can make various changes and improvements to the present disclosure in form and details, and these are all considered to fall within the scope of protection of the present disclosure.

Claims

1. Use of human serum albumin in the manufacture of a medicament for improving the skin condition of a subject.

2. The use according to claim 1, wherein the human serum albumin is human serum albumin prepared from human blood.

3. The use of claim 1, wherein the human serum albumin is young and undamaged human serum albumin.

4. The use of claim 3, wherein When compared to an endogenous human serum albumin preparation prepared from the plasma of young individuals, the young, undamaged human serum albumin exhibits at least one, e.g., two, three, and preferably four, of the following four properties: (1) a higher proportion of free sulfhydryl groups on the Cys-34 residue; (2) lower levels of advanced glycation end products (AGEs); (3) lower levels of carbonylation; and (4) lower levels of homocysteine.

5. The use of claim 3, wherein The young, undamaged human serum albumin exhibits at least one of the following four properties, for example, two, three, and preferably four: (1) the proportion of free thiol groups on the Cys-34 residue determined by the Ellman method is greater than 50%, for example, 70%, in particular 80%, preferably 90%, and more preferably greater than 95%; (2) the level of advanced glycation end products (AGE) determined by ELISA (CLOUD-CLONE Co., CEB353Ge) is lower than 60 μg / g protein, preferably lower than 40 μg / g protein, and more preferably lower than 30 μg / g protein; (3) the carbonyl level determined by the Protein Carbonyl Content Assay Kit is lower than 1.7 nmol / mg protein, preferably lower than 1.5 nmol / mg protein; (4) the level of homocysteine ​​determined by ELISA (Jianglai, JL10022) is lower than 5 nmol / g protein, preferably lower than 3.5 nmol / g protein, and more preferably lower than 2 nmol / g protein.

6. The use of claim 3, wherein the proportion of free sulfhydryl groups on the Cys-34 residue is greater than 80%, the AGE level is lower than 30 μg / g protein, the carbonyl level is lower than 1.5 nmol / mg protein, and the homocysteine ​​level is lower than 2 nmol / g protein.

7. The use according to any one of claims 1 to 6, wherein the drug further comprises sodium hyaluronate.

8. The use according to claim 7, wherein in the drug, the concentration of human serum albumin is 100 mg / ml, and the concentration of sodium hyaluronate is 1.00%.

9. The use according to any one of claims 1 to 8, wherein the drug further comprises another functional component.

10. The use according to any one of claims 1 to 9, wherein the functional component is selected from the group consisting of nicotinamide, α-lipoic acid, carnosine, and resveratrol.

11. The use according to any one of claims 1 to 10, wherein the drug is for mesodermal injection into a subject.

12. The use according to any one of claims 1 to 11, wherein the drug is in the form of an injection.

13. The use according to any one of claims 1 to 12, wherein the subject is a healthy subject in need of improving skin condition.

14. A skin repair preparation comprising: human serum albumin, and optionally a pharmaceutically acceptable carrier.

15. The skin repair preparation according to claim 14, wherein the human serum albumin is human serum albumin obtained from human blood.

16. The skin repair preparation of claim 14, wherein the human serum albumin is young, undamaged human serum albumin.

17. The skin repair preparation of claim 16, wherein When compared to an endogenous human serum albumin preparation prepared from the plasma of young individuals, the young, undamaged human serum albumin exhibits at least one, e.g., two, three, or preferably four, of the following four properties: (1) a higher proportion of free sulfhydryl groups at Cys-34 residues; (2) lower levels of advanced glycation end products (AGEs); (3) lower levels of carbonylation; and (4) lower levels of homocysteine.

18. The skin repair preparation of claim 16, wherein The young, undamaged human serum albumin exhibits at least one of the following four properties, for example, two, three, and preferably four: (1) the proportion of free thiol groups on the Cys-34 residue determined by the Ellman method is greater than 50%, for example, 70%, in particular 80%, preferably 90%, and more preferably greater than 95%; (2) the level of advanced glycation end products (AGE) determined by ELISA (CLOUD-CLONE Co., CEB353Ge) is lower than 60 μg / g protein, preferably lower than 40 μg / g protein, and more preferably lower than 30 μg / g protein; (3) the carbonyl level determined by the Protein Carbonyl Content Assay Kit is lower than 1.7 nmol / mg protein, preferably lower than 1.5 nmol / mg protein; (4) the level of homocysteine ​​determined by ELISA (Jianglai, JL10022) is lower than 5 nmol / g protein, preferably lower than 3.5 nmol / g protein, and more preferably lower than 2 nmol / g protein.

19. The skin repair preparation of claim 16, wherein the proportion of free sulfhydryl groups on the Cys-34 residue is greater than 80%, the AGE level is less than 30 μg / g protein, the carbonyl level is less than 1.5 nmol / mg protein, and the homocysteine ​​level is less than 2 nmol / g protein.

20. The skin repair formulation of any one of claims 14-19, wherein the formulation further comprises sodium hyaluronate.

21. The skin repair preparation according to any one of claims 14 to 20, wherein the concentration of human serum albumin in the preparation is 100 mg / ml and the concentration of sodium hyaluronate is 1.00%.

22. The skin repair formulation of any one of claims 14 to 21, wherein the formulation further comprises an additional functional component.

23. The skin repair formulation of any one of claims 14-22, wherein the functional component is selected from the group consisting of niacinamide, α-lipoic acid, carnosine, and resveratrol.

24. The skin repair formulation of any one of claims 14-23, wherein the formulation is for mesodermal injection into a subject.

25. The skin repair preparation according to any one of claims 14 to 24, wherein the preparation is in the form of an injectable dosage form.

26. The skin repair formulation of any one of claims 14-25, wherein the formulation is for use in healthy subjects in need of improved skin condition.

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