Cell growth factor emulsifiable paste for regeneration and repair of endometrial injury

Abstract

The cell growth factor emulsifiable paste for regeneration and repair of endometrial injury is composed of an emulsifiable paste matrix, an emulsifier and a cell growth factor solution, the cell growth factor solution is uniformly dispersed in the emulsifiable paste matrix, and the volume ratio of the emulsifiable paste matrix to the emulsifier to the cell growth factor solution is 10: 1: 1-1: 1: 1. The emulsifiable paste substrate is composed of one or more of plant-derived vegetable oil or hydrogenated vegetable oil, saturated hydrocarbon, ester chemically synthesized or semi-synthesized by higher fatty acid and higher fatty alcohol and a mixture thereof; the emulsifier is one or a combination of more of soap, higher fatty alcohol, polyol ester, fatty alcohol sulfate or sodium and polyoxyethylene ether derivatives, and the cell growth factor solution is formed by dispersing cell growth factors in a polyol solution which is in a liquid state at 25 DEG C without water and oxygen. The cell growth factor emulsifiable paste for regeneration and repair of endometrial injury overcomes the problems of poor physical and chemical stability, easy inactivation and the like of the cell growth factor, and gives full play to the regeneration and repair effect of the cell growth factor on endometrial injury.

Description

Technical Field

[0001] The invention relates to a pharmaceutical preparation of cell growth factor, in particular to a cell growth factor cream for regenerating and repairing endometrial damage. Background Art

[0002] Endometrial damage is the most common gynecological disease, such as endometrial damage and endometritis. It can occur in people of all ages and has great harm, such as affecting the quality of life and causing female infertility. After suffering from endometrial damage, women will experience symptoms such as increased vaginal discharge and vulvar itching, which seriously affect their normal life. If a woman suffers from endometrial damage during pregnancy, it will have a great impact on the development of the fetus, especially during childbirth, and the disease may be transmitted to the fetus, affecting the health of the fetus.

[0003] Cell growth factors are cytokines that have the activity of stimulating cell growth. Common cell growth factors include: epidermal growth factor (EGF), fibroblast growth factor (FGF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF, divided into IGF-1 and IGF-2), nerve growth factor (NGF), platelet-derived growth factor (PDGF), vascular endothelial cell factor (VEGF), transforming growth factor (TGF, divided into TGF-α and TGF-β), T cell growth factor, etc.

[0004] Cell growth factors are biological macromolecule drugs, which have problems such as poor physical and chemical stability and easy inactivation. Therefore, in order to form endometrial damage regeneration and repair preparations containing cell growth factors, their physical and chemical stability must be ensured to ensure their biological activity.

[0005] Creams refer to uniform semisolid preparations formed by dissolving or dispersing drugs in an emulsion-type matrix, which are divided into O / W or W / O creams. Creams are suitable for subacute, chronic, non-exudative skin lesions and skin pruritus, and are not suitable for erosion, ulcers, blisters and pustular diseases. Compared with O / W creams, W / O creams have better stability, and the water is in the internal phase, which is not easy to deteriorate and evaporate. Therefore, various physical and chemical parameters are good, and they are commonly used for drug administration applications of skin and mucosa. Although creams have many advantages in the treatment of mucosal wounds, cell growth factors are biomacromolecule drugs with poor physical and chemical stability and easy inactivation. There are no reports on preparations carrying cell growth factors for the treatment of endometrial damage. Therefore, the preparation of safe, stable and efficient preparations carrying cell growth factors is a limiting bottleneck for the use of FGF in the treatment of endometrial damage. Summary of the invention

[0006] The purpose of the present invention is to overcome the shortcomings of the prior art - the lack of a safe and stable preparation of cell growth factors for the treatment of endometrial damage, to provide a cream that efficiently carries cell growth factors, to break through the restrictive bottleneck of cell growth factors in the treatment of endometrial damage, and at the same time meet the safety, effectiveness, convenience and economy requirements of clinical treatment.

[0007] The inventors have found that cell growth factors have poor physical and chemical stability and are easily oxidized and inactivated. To this end, the inventors have prepared a highly dispersible cream of stable cell growth factors after a large number of experiments, achieving the purpose of safety, effectiveness, convenience and economy, breaking through the restrictive bottleneck of cell growth factors in the treatment of endometrial damage, and giving full play to the therapeutic effect of cell growth factors on body diseases, especially the therapeutic effect of cell growth factors on endometrial damage.

[0008] The cell growth factor cream for regeneration and repair of endometrial damage of the present invention is composed of a cream base, an emulsifier and a cell growth factor solution, wherein the cell growth factor solution is uniformly dispersed in the cream base;

[0009] The volume ratio of the above-mentioned cream base, emulsifier and cell growth factor solution is 10:1:1 to 1:1:1;

[0010] The above-mentioned cream base is composed of one or more combinations of vegetable oil or hydrogenated vegetable oil of plant origin, saturated hydrocarbons, chemically synthesized or semi-synthetic esters of higher fatty acids and higher fatty alcohols, and mixtures thereof;

[0011] The above-mentioned emulsifier is one or a combination of soaps, higher fatty alcohols, polyol esters, fatty alcohol sulfates or sodiums, and polyoxyethylene ether derivatives;

[0012] The cell growth factor solution is formed by dispersing the cell growth factor in a polyol that is liquid at 25° C. without water or oxygen.

[0013] The above-mentioned cream is used to achieve the regeneration and repair effect of cell growth factors on endometrial damage;

[0014] The cell growth factor is one or a combination of epidermal growth factor, fibroblast growth factor, vascular endothelial growth factor, and transforming growth factor;

[0015] The volume ratio of the above-mentioned cream matrix to the cell growth factor solution is 4:1:1 to 2:1:1;

[0016] The volume ratio of the above-mentioned cream base, emulsifier and cell growth factor solution is 3:1:1;

[0017] The saturated hydrocarbon is one or a combination of vaseline, solid paraffin, and liquid paraffin;

[0018] The chemically synthesized or semi-synthesized esters of higher fatty acids and higher fatty alcohols and mixtures thereof are one or a combination of stearic acid glyceryl ester, lanolin, beeswax and spermaceti;

[0019] The synthetic / semi-synthetic oils are one or a combination of squalane, lanolin derivatives, and silicones;

[0020] The above-mentioned polyol which is liquid at 25°C is one or a combination of polyethylene glycol 200-800, glycerol, and propylene glycol;

[0021] A method for preparing the above-mentioned cell growth factor cream for regeneration and repair of endometrial damage comprises the following steps:

[0022] a. Take 100 ml of polyol that is liquid at 25°C and heat it to 100°C to remove oxygen, transfer it into a stoppered glass bottle, saturate the inner space of the glass bottle with nitrogen, cool it to 25°C, add 2 mg of cell growth factor lyophilized powder and mix well to form a cell growth factor solution;

[0023] b. Take the cream base and emulsifier according to the prescribed volume ratio, heat the cream base to melt first, then add the emulsifier to the cream base and mix well, cool to 25°C, add the cell growth factor solution prepared in step a, mix well to form a cell growth factor cream for regeneration and repair of endometrial damage, and store in a cool and sealed place.

[0024] One or a combination of antioxidants, antibacterial agents and pH regulators commonly used in pharmacy are added to the cell growth factor cream for regeneration and repair of endometrial damage.

[0025] The cell growth factor cream of the present invention has the following advantages: 1. The cell growth factor is made into a semisolid preparation, which not only enhances the stability of the cell growth factor but also facilitates application in local pharmaceutical treatment; 2. The cell growth factor is highly dispersed in a water-free cream matrix to avoid contact of the cell growth factor with external light, oxygen, etc., thereby ensuring its stability; 3. The cream is simple to prepare and easy to use, which is conducive to achieving the purpose of rapid and long-term regeneration and repair of endometrial damage. DETAILED DESCRIPTION

[0026] The specific embodiments of the present invention will be described in detail below. It should be noted that the technical features or combinations of technical features described in the following embodiments should not be considered isolated, and they can be combined with each other to achieve better technical effects.

[0027] Example 1 Preparation of cell growth factor preparation for regeneration and repair of endometrial damage

[0028] According to the component ratios in Table 1, the cream preparation of the experimental group was prepared using the following steps:

[0029] a. Take 100 ml of polyol that is liquid at 25°C and heat it to 100°C to remove oxygen, transfer it into a stoppered glass bottle, saturate the inner space of the glass bottle with nitrogen, cool it to 25°C, add 2 mg of cell growth factor lyophilized powder and mix well to form a cell growth factor solution;

[0030] b. Take the cream base and emulsifier according to the volume ratio designed in Table 1, first heat the cream base to melt, then add the emulsifier to the cream base and mix well, cool to 25°C, add the cell growth factor solution prepared in step a, mix well to form a cell growth factor cream for regeneration and repair of endometrial damage, and store in a cool and sealed place.

[0031] The control group preparation was prepared with reference to the experimental group preparation according to the component ratios in Table 1. Each experimental group was configured according to the components and ratios within the protection scope of the claims of this application, while each control group had a component replaced by another component or a component value beyond the protection scope of the claims of this application.

[0032] Table 1 Preparation of experimental and control groups

[0033]

[0034] Note: * represents that the component or value of the item is significantly different from that of the experimental group; / represents that the component is missing or the value is 0; Control groups 1 and 2 are O / W creams; IGF-2 is insulin-like growth factor 2; IL-2 is T cell growth factor, also known as interleukin-2; EGF is epidermal growth factor; FGF is fibroblast growth factor; VEGF is vascular endothelial growth factor; TGF is transforming growth factor.

[0035] Example 2 Effect of the cell growth factor preparation on regeneration and repair of endometrial damage

[0036] (1) Establishment of endometrial injury model animals

[0037] Eight-week-old female Sprague-Dawley rats (230-250 g) were used to establish a rat model of endometrial injury. Rats were housed in standard cages at a temperature of 22 ± 1 °C, a relative humidity of 50 ± 1%, a light-dark cycle of 12 / 12 h, and adequate food and drinking water were provided daily. The endometrial injury model was established by injecting anhydrous ethanol into the uterine cavity. In brief, rats were anesthetized by intraperitoneal injection of 2% sodium pentobarbital (0.3 mL / 100 g), placed in a supine position, the lower abdomen was prepared, disinfected with 75% alcohol and iodine, and a longitudinal incision was made along the midline of the abdomen to open the abdominal cavity and expose the uterus. Two vascular clamps were used to clamp the right uterus proximal to about 0.5 cm above the bifurcation and the distal third, and anhydrous ethanol was slowly injected into the uterine cavity for 5 minutes. Then the vascular clamps were loosened, the ethanol was gently squeezed to remove it, and the damaged area was rinsed with saline to further remove the residual ethanol. Samples were taken for observation. After confirming that the endometrial injury model was successfully established, the abdominal cavity was rinsed with 37 °C saline and the abdominal cavity was sutured. Within three days after surgery, rats were routinely injected with sodium penicillin 200,000U / kg into the thigh muscle every day to prevent wound infection.

[0038] (2) Evaluation of the therapeutic effects of each group of preparations on model animals

[0039] Using the rat endometrial injury model established above, each group of preparations was injected in situ into the damaged endometrium, and the repair effect of each group of creams on endometrial damage was compared and analyzed, and the animal fur color and diet were observed. On the 8th day after administration, the rats were euthanized, and the uterine tissues were collected for pathological sections. The repair effect of the rat endometrium was determined by HE staining, TUNEL cell apoptosis, and immunohistochemistry; transmission electron microscopy (TEM) was used to observe the electron microscopic specimens of the endometrium in each experimental group, and the tissue morphological changes of the endometrial damage site after the application of different groups of preparations were compared.

[0040] Based on the above results, the double-blind method was used to evaluate the regeneration and repair effect of each group of preparations on endometrial damage, and a total score (full score 100 points) was given, and the higher the score, the better the overall effect. The experimental results of each group of preparations are shown in Table 2.

[0041] Table 2 Regeneration and repair effects of the experimental and control group preparations on endometrial damage

[0042]

[0043]

[0044] From the experimental results in Table 2, it can be seen that the cream in the experimental group has a good regeneration and repair effect on endometrial damage, especially in experimental group 7, with good indicators, which verifies that the experimental group has a good regeneration and repair effect on endometrial damage in rats. Compared with the experimental group, the regeneration and repair effect of the control group preparation on endometrial damage is significantly poorer, especially in control groups 8 and 9, with poor evaluation indicators.

[0045] As can be seen from the above, each experimental group has a good regeneration and repair effect on rat endometrial damage, and can be used as a regeneration and repair preparation for endometrial damage. Therefore, the cell growth factor cream for regeneration and repair of endometrial damage of the present invention exerts the regeneration and repair effect of cell growth factors on endometrial damage, and has a good application prospect.

[0046] The above detailed description is a specific description of a feasible embodiment of the invention, and the embodiment is not intended to limit the patent scope of the present invention. Any equivalent implementation or change that does not deviate from the present invention should be included in the patent scope of the present invention. In addition, those skilled in the art can also make various modifications, additions and substitutions in other forms and details within the scope and spirit disclosed in the claims of the present invention. Of course, these various modifications, additions and substitutions made according to the spirit of the present invention should be included in the scope of protection claimed by the present invention.

Claims

1. A cell growth factor cream for regeneration and repair of endometrial damage, wherein the main characteristics are as follows: the cream is composed of a cream base, an emulsifier and a cell growth factor solution, and the cell growth factor solution is uniformly dispersed in the cream base; the volume ratio of the cream base, the emulsifier and the cell growth factor solution is 10:1:1 to 1:1:1; the cream base is composed of one or more combinations of plant oil or hydrogenated plant oil, saturated hydrocarbons, chemically synthesized or semi-synthetic esters of higher fatty acids and higher fatty alcohols and mixtures thereof; the emulsifier is one or more combinations of soaps, higher fatty alcohols, polyol esters, fatty alcohol sulfates or sodiums, and polyoxyethylene ether derivatives; the cell growth factor solution is formed by dispersing the cell growth factor in a polyol that is liquid at 25°C in the absence of water and oxygen; the cream is used to achieve the regeneration and repair effect of the cell growth factor on endometrial damage.

2. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The cell growth factor is one or a combination of epidermal growth factor, fibroblast growth factor, vascular endothelial growth factor and transforming growth factor.

3. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The volume ratio of the cream matrix to the cell growth factor solution is 4:1:1 to 2:1:

1.

4. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The volume ratio of the cream base, the emulsifier and the cell growth factor solution is 3:1:

1.

5. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The saturated hydrocarbon is one or a combination of vaseline, solid paraffin and liquid paraffin.

6. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The chemically synthesized or semi-synthesized esters of higher fatty acids and higher fatty alcohols and their mixtures are one or a combination of stearic acid glyceryl ester, lanolin, beeswax and spermaceti.

7. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The synthetic or semi-synthetic oil is one or a combination of squalane, lanolin derivatives and silicone.

8. The cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The polyol which is liquid at 25° C. is one or a combination of polyethylene glycol 200-800, glycerol and propylene glycol.

9. A method for preparing the cell growth factor cream for regeneration and repair of endometrial damage according to claim 1, characterized in that: The steps include: a. Take 100 ml of polyol that is liquid at 25°C and heat it to 100°C to remove oxygen, transfer it into a stoppered glass bottle, saturate the inner space of the glass bottle with nitrogen, cool it to 25°C, add 2 mg of cell growth factor lyophilized powder and mix well to form a cell growth factor solution; b. Take the cream base and emulsifier according to the prescribed volume ratio, heat the cream base to melt first, then add the emulsifier to the cream base and mix well, cool to 25°C, add the cell growth factor solution prepared in step a, mix well to form a cell growth factor cream for regeneration and repair of endometrial damage, and store in a cool and sealed place.

10. A cell growth factor cream for regeneration and repair of endometrial damage according to claim 9, characterized in that: One or a combination of antioxidants, antibacterial agents and pH regulators commonly used in pharmacy are added to the cell growth factor cream for regeneration and repair of endometrial damage.

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