CN109820753B - Essence for promoting subcutaneous fat regeneration and preparation process thereof - Google Patents
Essence for promoting subcutaneous fat regeneration and preparation process thereof Download PDFInfo
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Abstract
The invention relates to the field of skin anti-aging nursing products, in particular to essence for promoting subcutaneous fat regeneration and a preparation process thereof, wherein the formula comprises the following components in parts by mass: 30.01-1.00% of SIS, 4315420.01-1.00% of SB, 0.05-1.5% of sodium hyaluronate, 0.5% of pyrrolidone carboxylate, 0.01-1% of olive oil, 1-5% of glycerol, 0.05-2.00% of Sepigel 3050.05-2.00% of deionized water and the balance of deionized water, wherein the sum of the components is 100%; the SIS3 is a novel specific Smad3 inhibitor, can inhibit phosphorylation of Smad3 to inhibit TGF-beta and activin signals, SB-431542 is a transforming growth factor beta Receptor (TGF-beta Receptor) inhibitor, SIS3 and SB431542 are both fat-soluble, have molecular weights less than 800, and easily penetrate through a skin stratum corneum to enter a basal layer and a dermis layer of the skin.
Description
Technical Field
The invention relates to the field of anti-aging skin care products, in particular to essence for promoting subcutaneous fat regeneration and a preparation process thereof.
Background
With the increasing standard of living, people pay more and more attention to health, especially to skin care. The skin tissue is the largest tissue organ of the human body, covers the surface of the human body and blocks the damage effect of all harmful substances in the nature on living organisms. However, with the increase of age, the functions of skin tissues and organs are gradually aged, and from the aesthetic point of view, the enthusiasm of people for life is seriously influenced; from the physiological point of view, the effective protection of internal organs of a human body is influenced, the skin aging is a comprehensive problem, generally, with the continuous increase of the age, the problems of subcutaneous fat atrophy, collagen fiber and elastic fiber degeneration and the like are continuously aggravated, the thickness and the elasticity of the skin are reduced, and finally, the skin gradually loosens and sags. The atrophy of subcutaneous fat is an important reason for the reduction of the thickness and elasticity of skin, and how to effectively increase the content of the subcutaneous fat is one of the main means for solving the problems of skin shriveling and sagging. The method is also the theoretical basis for filling the face with autologous fat for filling the facial depressions, removing wrinkles and solving the facial skin sagging in medical cosmetology at present. However, the autologous fat face filling belongs to an operation project, certain operation risk and postoperative infection risk exist, and meanwhile, due to the fact that the survival rate of transplanted autologous fat has certain problems, the problem that the face is not symmetrical after the autologous fat face filling is easily caused is solved. Therefore, the most ideal method is to find out the fundamental reason by deeply researching the physiological process of the subcutaneous lipoatrophy, and to re-induce the regeneration of fat cells by intervention of cytology level so as to solve the problem of the subcutaneous lipoatrophy.
The skin of the infant has a large amount of fat, so that the skin is bright and plump, but with the age, the subcutaneous fat slowly disappears, the subcutaneous fat is completely replaced by the fibrotic cells, and the skin is characterized by skin sag, sagging and the like by the old. Dermal fibroblasts (dermal fibroblasts) in the dermis layer of the skin have the capability of differentiating into connective tissue cells such as fat cells and fibroblasts, and the physiological condition of the body gradually changes with the age, so that the capability of the dermal fibroblasts (dermal fibroblasts) for differentiating into fat cells is weakened, and most of the dermal fibroblasts are differentiated into fibroblasts, which is the main reason of the gradual reduction of subcutaneous fat with the age. Recent studies have shown that the differentiation of dermal fibroblasts (dermal fibroblasts) is regulated by TGF beta signaling pathway, and as the body ages, TGF beta level increases, TGF beta related signaling pathway in dermal fibroblasts (dermal fibroblasts) is activated, inhibiting its ability to differentiate into adipocytes. Cytological level and animal model research show that the TGF beta cell signal pathway of dermal fibroblasts (dermal fibroblastits) is inhibited, and the differentiation of the dermal fibroblasts into adipocytes can be effectively promoted.
Disclosure of Invention
In order to solve the problems of safety risk, postoperative infection risk and postoperative asymmetry of the face in skin aging in the prior medical cosmetology by an operation mode, the invention provides essence for promoting subcutaneous fat regeneration, which has the technical scheme as follows: the essence for promoting the regeneration of subcutaneous fat is characterized by comprising the following components in parts by mass: 30.01-1.00% of SIS, 4315420.01-1.00% of SB, 0.05-1.5% of sodium hyaluronate, 0.5% of pyrrolidone carboxylate, 0.01-1% of olive oil, 1-5% of glycerol, 0.05-2.00% of Sepigel 3050.05-2.00% of deionized water and the balance of deionized water, wherein the sum of the components is 100%;
the preferable combination formula of the invention comprises the following components in percentage by mass: 30.1% of SIS, 4315420.1% of SB, 0.1% of sodium hyaluronate, 0.5% of pyrrolidone carboxylate, 1% of olive oil, 5% of glycerol, 3052.00% of Sepigel and the balance of deionized water, wherein the sum of the components is 100%;
the performance and the matching function effect of each component in the formula are as follows:
SIS 3: is a novel specific Smad3 inhibitor, inhibits TGF-beta and activin signals by inhibiting phosphorylation of Smad3, and does not affect MAPK/p38, ERK or PI3-kinase signal channels;
SB 431542: is a potent, ATP-competitive ALK5 Inhibitor (IC)50=94 nM) and acts on ALK4 and ALK7, but does not affect p38 kinase activity. In addition, SB431542 inhibits TGF-. beta.and activin-induced phosphorylation of Smad 2. SB431542 inhibits TGF-. beta.induced apoptosis and growth inhibitory effects, and effectively inhibits TGF-. beta.tumor promotion effects including cell migration, cell invasion and VEGF secretion. SB431542 specifically blocks Smad signaling, reducing gene expression associated with fibrosis and cancer;
sodium hyaluronate: the hyaluronic acid gel is an inherent component in a human body, is a glucuronic acid, has no species specificity, is widely present in cytoplasm and intercellular substance in tissue organs such as placenta, amniotic fluid, crystalline lens, articular cartilage, skin dermis and the like, plays a role in lubricating and nourishing cells and cell organs contained in the hyaluronic acid, and provides a microenvironment for cell metabolism at the same time;
pyrrolidone carboxylic acid salt: the amino acid derivative is a substance naturally existing in the skin, is soluble in water and ethanol but insoluble in oil, has stronger hygroscopicity, and can absorb moisture from the air, and the amino acid derivative is used as a humectant in cosmetics, and has stronger moisturizing capability than traditional humectants such as glycerol, propylene glycol and sorbitol;
olive oil: the health care wine is praised in the western world as 'liquid gold', 'vegetable oil queen', 'mediterranean nectar', because the health care wine has excellent natural health care efficacy, beauty treatment efficacy and moisture retention efficacy;
glycerol: the chemical name is glycerol, which has strong water absorption. Normally we use glycerin with 20% moisture. The skin can be moistened by applying the skin care cream on the skin in winter, so that the skin is not chapped;
sepigel 305: the emulsion thickening, emulsifying and stabilizing agent has the advantage of high swelling speed, can be instantly prepared into gel, and is a new material for producing cream at normal temperature and high temperature without neutralization. It can emulsify various oils regardless of their HLB values, and can disperse pigments and physical sunscreens. The emulsion can be compounded with other traditional emulsifiers or used independently for preparing various skin-care cream, gel and other products, and the cream prepared from the emulsion has bright appearance, excellent touch, softness and smoothness.
A preparation process of essence for promoting subcutaneous fat regeneration is characterized by comprising the following steps:
1. firstly, adding sodium hyaluronate, pyrrolidone carboxylate, Sepigel305, glycerol and deionized water into a reaction tank, starting stirring, heating to 75-80 ℃, and uniformly stirring and dispersing;
2. and (3) continuing stirring and cooling to below 37 ℃ in the first step, dissolving SIS3 and SB431542 in olive oil, adding into a reaction tank, homogenizing for 2-3 minutes, and completely stirring and dissolving to finish the process.
The invention has the beneficial effects that: the SIS3 is a novel specific Smad3 inhibitor, can inhibit phosphorylation of Smad3 to inhibit TGF-beta and activin signals, the SB-431542 is a transforming growth factor beta Receptor (TGF-beta Receptor) inhibitor, the SIS3 and the SB431542 are both fat-soluble and have molecular weights less than 800, and easily enter a basal layer and a dermis layer of skin through a skin cuticle layer.
Drawings
FIG. 1 is a chemical structural diagram of SIS3 in the present invention;
FIG. 2 is a chemical structural diagram of SB431542 in the present invention;
FIG. 3 is a graph comparing the skin fat thickness of experimental groups of mice in the section of the detailed description of the invention;
Detailed Description
Specific example 1: firstly, adding 0.05% of sodium hyaluronate, 0.5% of pyrrolidone carboxylate, 0.05% of Sepigel305, 1% of glycerol and 98.38% of deionized water into a reaction tank, starting stirring, heating to 75-80 ℃, and uniformly stirring and dispersing; and then, continuously stirring and cooling to the temperature lower than 37 ℃, dissolving 0.01% of SIS3 and 0.01% of SB431542 in 0.01% of olive oil, adding into a reaction tank, homogenizing for 2-3 minutes, and completely stirring and dissolving to finish.
Specific example 2: firstly, 1.5 percent of sodium hyaluronate, 0.5 percent of pyrrolidone carboxylate, 2.00 percent of Sepigel305, 5 percent of glycerol and 88 percent of deionized water are added into a reaction tank, stirred, heated to 75-80 ℃, and uniformly stirred and dispersed; and then, continuously stirring and cooling to the temperature lower than 37 ℃, dissolving 1% of SIS3 and 1% of SB431542 in 1% of olive oil, adding into the reaction tank, homogenizing for 2-3 minutes, stirring and dissolving completely, and finishing.
Specific example 3: firstly, 0.1% of sodium hyaluronate, 0.5% of pyrrolidone carboxylate, 2.00% of Sepigel305, 5% of glycerol and 92.2% of deionized water are added into a reaction tank, stirring is started, heating is carried out to 75-80 ℃, and stirring and dispersing are carried out uniformly; and then, continuously stirring and cooling to the temperature lower than 37 ℃, dissolving 0.1% of SIS3 and 0.1% of SB431542 in 1% of olive oil, adding into the reaction tank, homogenizing for 2-3 minutes, stirring and completely dissolving, and finishing.
The invention is further illustrated below with reference to the formulation of specific embodiments:
SIS3 and SB431542 are core effective components for promoting subcutaneous fat regeneration in the invention, respectively inhibit signals at different nodes of a TGF beta signal path, and are configured according to different concentrations according to experimental design, and the specific experimental group formula is as follows:
the performance test method and the effect of each experimental group are as follows:
1) C57/BL6 mice of 48 weeks old are taken, 100ul of 1% pentobarbital sodium is injected into the mice through micro-veins, the hairs of the mice are cut short after the mice are completely anesthetized, then 8% sodium sulfide water solution is dipped by cotton balls, a thin layer is coated on the trunk below the neck of the mice, the fallen hairs are washed away by warm water after 2-3 minutes, and the hairs are wiped by gauze;
2) the group after the leg hairs was divided into 7 groups of 5 mice each, and the mice in each group were individually numbered A, B, C, D, E. Group 0 mice were treated with the serum of experimental group 0, group 1 with the serum of experimental group 1, and so on for each experimental group. The specific treatment method comprises the following steps: weighing 0.2g of essence, uniformly smearing the essence on the naked parts of the mouse trunk, smearing the essence once every 24 hours for 7 days;
3) after 7 days, all mice were sacrificed by injecting 400ul of 1% sodium pentobarbital 350ul, skin tissue blocks of 0.5cm x 0.3cm were excised from the median abdominal region, skin tissue sections were prepared, and then HE-stained, and the thickness of the adipose layer of the tissue sections was observed under a microscope. The method comprises the following specific steps:
1. removing hair from skin tissue at the median part of abdomen of mouse, placing into PBS (pH 7.2) to thoroughly rinse residual blood
2. The rinsed tissue was fixed overnight in 4% paraformaldehyde (1:5 ratio soak)
3. Washing the fixed tissue twice with PBS for 30min each time
4. Tissue dehydration 30% ethanol → 50% ethanol → 75% ethanol (at 4 deg.C overnight) → 85% ethanol → 95% ethanol → 100% ethanol (each 30min)
5. Transparent xylene transparent for 3 × 20min (the specific time is set according to the transparency degree, the skin of the mouse is transparent for 10min,
when the tissue is transparent, if the tissue is found to be partially white and cloudy, the tissue may be dehydrated and not completely removed)
6. Waxing 4X 30min (wax is changed once every 30min, and four times to make paraffin soak in tissue for supporting function)
7. Embedding (attention is paid to tissue placement and transverse cutting, longitudinal cutting and label recording), and repairing the paraffin (embedded as soon as the paraffin is fully solidified
Hot wax block not frozen immediately)
8. Slicing (5-7um), spreading on clean glass slide (40-45 deg.C), and baking (about 62 deg.C, oven drying)
HE staining xylene I (10min) → xylene II (5min) → xylene ethanol =1:1(2min) → 100% ethanol → 95% ethanol (2min) → 85% ethanol (1min) → 70% ethanol (1min) → 50% ethanol (1min) → water (1min) → hematoxylin staining solution (1min) → running tap water (3min) → 50% ethanol (1min) → 70% ethanol (1min) → 85% ethanol (1min) → 95% ethanol (1min) → eosin staining solution (1min) → 95% ethanol (1min) → 100% ethanol (1min) → xylene I (2min) → xylene II (2min)
10. Placing a microscopic micrometer beside the tissue section, observing and photographing under a microscope, and measuring the thickness of the adipose tissues in the section by taking the microscopic micrometer as a reference;
4) data statistics
The mean value and variance of the fat thickness measured in each group are plotted and compared, and the result is shown in the attached figure 3 of the specification,
when the effective concentration of SIS3 and SB431542 reached 0.1% (group 7), the increase in fat in skin tissue was effectively promoted in C57/BL6 mice at 48 weeks of age, and the increased fat thickness was about 3 times greater than that in the control group (group 0) to which no TGF beta signaling pathway inhibitor was added.
Through the above studies, it was found that the mode of skin administration is suitable for the simultaneous application of two TGF beta signaling pathway inhibitors, SIS3 and SB431542, to the regeneration of skin fat, and the differentiation of dermal fibroblasts into adipocytes can be effectively promoted. The research result shows that SIS3 and SB431542 have potential value as external medicines or skin care products, and the skin condition is improved by applying the skin condition-improving essence to the regeneration of subcutaneous fat.
Claims (3)
1. The essence for promoting the regeneration of subcutaneous fat is characterized by comprising the following components in parts by mass: SIS3, molecular formula: c28H28ClN3O3The chemical structural formula is as follows:
0.01-1.00%, SB431542, molecular formula: c22H16N4O3The chemical structural formula is as follows:
0.01-1.00 percent of sodium hyaluronate, 0.05-1.5 percent of sodium hyaluronate, 0.5 percent of pyrrolidone carboxylate, 0.01-1 percent of olive oil, 1-5 percent of glycerol, 0.05-2.00 percent of Sepigel 305and the balance of deionized water, wherein the sum of the components is 100 percent.
2. The essence for promoting the regeneration of subcutaneous fat according to claim 1, wherein the essence comprises the following components in parts by mass: 30.1 percent of SIS, 4315420.1 percent of SB, 0.1 percent of sodium hyaluronate, 0.5 percent of pyrrolidone carboxylate, 1 percent of olive oil, 5 percent of glycerol, 3052.00 percent of Sepigel and the balance of deionized water, wherein the sum of the components is 100 percent.
3. A process for preparing the essence for promoting the regeneration of subcutaneous fat according to claim 1, comprising the steps of:
(1) Firstly, adding sodium hyaluronate, pyrrolidone carboxylate, Sepigel305, glycerol and deionized water into a reaction tank, starting stirring, heating to 75-80 ℃, and uniformly stirring and dispersing;
(2) And continuing to stir and cool to a temperature lower than 37 ℃ in the first step, then dissolving SIS3 and SB431542 in olive oil, adding into a reaction tank, homogenizing for 2-3 minutes, stirring and dissolving completely, and finishing.
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| CN108300686A (en) * | 2017-01-13 | 2018-07-20 | 云南美颜美龄生物科技有限公司 | For preventing, delaying or micromolecular compound/combination of reverse both, tissue, organ, body aging, product and application thereof |
| CN108451790A (en) * | 2018-05-30 | 2018-08-28 | 海默斯（重庆）医学生物技术有限公司 | A kind of moisturizing essence of the keratin containing hydrophily and preparation method thereof |
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| CN108451790A (en) * | 2018-05-30 | 2018-08-28 | 海默斯（重庆）医学生物技术有限公司 | A kind of moisturizing essence of the keratin containing hydrophily and preparation method thereof |
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