Preparation process of minoxidil mixed solution

Abstract

The invention discloses a preparation process of minoxidil mixed liquor, and relates to the technical field of pharmaceutical preparations. When minoxidil mixed liquid is prepared, soybean lecithin and ginsenoside are mixed and dried, and a phospholipid-ginsenoside film is prepared; the preparation method comprises the following steps: coating minoxidil with a phospholipid-ginsenoside film, so as to prepare a minoxidil-ginsenoside carrier solution; the preparation method comprises the following steps: extracting the isatis herb with deionized water to prepare an isatis herb extract; the preparation method comprises the following steps: loading a Umbrella grass extract into human umbilical cord mesenchymal stem cells to prepare a stem cell-loaded Umbrella grass extract; and mixing the minoxidil-ginsenoside carrier solution, the stem cell loaded Umbra grass extract and deionized water, so as to prepare a minoxidil mixed solution. The minoxidil mixed solution prepared by the invention is used for treating androgen alopecia, has an excellent treatment effect, and greatly reduces the adverse reactions such as coarse hair, scalp itching, rash and desquamation caused by single use of minoxidil.

Description

Technical Field

[0001] The invention relates to the technical field of pharmaceutical preparations, in particular to a preparation process of a minoxidil mixed solution. Background Art

[0002] Hair loss seriously affects people's physical and mental health. There are many types of hair loss. According to the current clinical medical classification, it can generally be divided into senile alopecia, chemotherapy alopecia, androgenic alopecia, traction alopecia, alopecia areata, drug-induced alopecia and scarring alopecia. At present, due to mental stress or lifestyle reasons, androgenic alopecia has the highest incidence, followed by alopecia areata. Androgenic alopecia is a chronic progressive disease, and the incidence rate in men is higher than that in women in the natural environment. Androgenic alopecia is characterized by the gradual miniaturization of hair follicles and the thinning of hair shafts. It is a multifactorial disease caused by the interaction of multiple factors such as genes, androgens, age and environment.

[0003] Minoxidil is a drug approved by the U.S. Food and Drug Administration for the treatment of hair loss. It can increase blood flow from the skin to the scalp by activating potassium channels, providing hair follicles with good blood supply and promoting hair follicle growth. However, minoxidil can cause adverse reactions such as rough hair, itchy scalp, rash, and desquamation. In order to reduce the adverse reactions caused by patients using minoxidil and improve the treatment effect, it is necessary to make improvements based on existing technologies. Summary of the invention

[0004] The object of the present invention is to provide a process for preparing a minoxidil mixed solution to solve the problems existing in the prior art.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: A preparation process of a minoxidil mixed solution, comprising the steps of wrapping minoxidil with a phospholipid-ginsenoside membrane to obtain a minoxidil-ginsenoside transfersome solution; loading a Usma grass extract into human umbilical cord mesenchymal stem cells to obtain a stem cell-loaded Usma grass extract; and mixing the minoxidil-ginsenoside transfersome solution, the stem cell-loaded Usma grass extract, and deionized water to obtain a minoxidil mixed solution. The phospholipid-ginsenoside membrane is prepared by mixing and drying soybean lecithin and ginsenoside; The Usma grass extract is prepared by extracting Usma grass with deionized water.

[0006] As an optimization, the preparation process of the minoxidil mixed solution includes the following preparation steps: (1) Add minoxidil and Tween 80 in a mass ratio of 2:(0.12-0.13) to deionized water 8-10 times the mass of minoxidil, stir at 10-30°C and 300-500 r / min for 16-20 min, add phospholipid-ginsenoside membrane 4-5 times the mass of minoxidil, heat to 50-60°C, continue stirring for 1-2 h, cool to room temperature and let stand for 1-2 h to obtain a transfersome suspension; ultrasonically treat the transfersome suspension at 0-2°C for 12-14 min, filter through a 0.22 µm microporous filter membrane to obtain a minoxidil-ginsenoside transfersome solution; (2) Human umbilical cord mesenchymal stem cells, trypsin and PBS solution were mixed and ultrasonically dispersed for 2-3 minutes, placed in a centrifuge tube, and centrifuged at 900 r / min for 4-6 minutes. The resulting precipitate was the stem cell membrane; Usma grass extract, stem cell membrane and PBS solution were mixed in a mass ratio of 1:(3-4):(40-50), placed in a centrifuge tube, and centrifuged at 900 r / min for 7-8 minutes. The precipitate was the stem cell-loaded Usma grass extract; minoxidil-ginsenoside transfersome solution, stem cell-loaded Usma grass extract and deionized water were evenly mixed to prepare a minoxidil mixed solution.

[0007] As an optimization, the preparation method of the phospholipid-ginsenoside film in step (1) is as follows: soybean lecithin and ginsenoside are added in a mass ratio of 7:2 to a mixed solvent of 4 to 5 times the mass of soybean lecithin, ultrasonically dispersed at room temperature for 10 to 12 minutes, and dried at 20 to 30° C. for 5 to 6 hours under vacuum conditions to obtain a phospholipid-ginsenoside film.

[0008] As an optimization, the preparation method of the mixed solvent is: chloroform and methanol are evenly mixed in a mass ratio of 10:1 to prepare a mixed solvent.

[0009] As an optimization, the preparation method of the Usma grass extract in step (2) is: uniformly mix the Usma grass powder and deionized water in a mass ratio of 1: (40~50), stir at 100°C, 300~500r / min for 2~3h, filter with filter cloth, uniformly mix the filter residue and deionized water in a mass ratio of 1: (20~24), stir at 100°C, 300~500r / min for 1h, filter with filter cloth, mix the two filtrates, centrifuge at 4500r / min for 9~11min, and dry the supernatant at 50~60°C under vacuum conditions for 4~5h to obtain the Usma grass extract.

[0010] As an optimization, the Usma grass powder is prepared by crushing the Usma grass into Usma grass powder with a particle size of less than 3 mm using a wall breaking machine.

[0011] As an optimization, the mass ratio of the human umbilical cord mesenchymal stem cells, trypsin, and PBS solution in step (2) is 1:(0.2~0.3):(80~100).

[0012] As an optimization, the model of the PBS solution in step (2) is CD433577.

[0013] As an optimization, the mass ratio of the minoxidil-ginsenoside transfersome solution, stem cell-loaded Usma grass extract, and deionized water in step (2) is 1:(0.2~0.3):(3~4).

[0014] Compared with the prior art, the beneficial effects achieved by the present invention are: When preparing the minoxidil mixed solution, the present invention comprises the following steps: mixing and drying soybean lecithin and ginsenoside to prepare a phospholipid-ginsenoside membrane; wrapping minoxidil with the phospholipid-ginsenoside membrane to prepare a minoxidil-ginsenoside transfer body solution; extracting Usma grass with deionized water to prepare a Usma grass extract; loading the Usma grass extract into human umbilical cord mesenchymal stem cells to prepare a stem cell-loaded Usma grass extract; and mixing the minoxidil-ginsenoside transfer body solution, the stem cell-loaded Usma grass extract and deionized water to prepare the product.

[0015] First, soybean lecithin and ginsenosides are mixed and dried to prepare a phospholipid-ginsenoside membrane; minoxidil is wrapped with the phospholipid-ginsenoside membrane to prepare a minoxidil-ginsenoside transfersome solution; minoxidil is a drug approved by the US Food and Drug Administration for the treatment of hair loss, which can increase the blood flow from the skin to the scalp by activating potassium channels, provide good blood supply to the hair follicles and promote hair follicle growth. However, minoxidil can cause adverse reactions such as rough hair, itchy scalp, rash, and desquamation; ginsenosides have anti-inflammatory effects and a steroid structure similar to cholesterol, which has a strong direct interaction with phospholipids and can be tightly inserted into the phospholipid bilayer; encapsulating minoxidil in ginsenosides can improve the transdermal absorption rate of minoxidil, while reducing the irritation of minoxidil to the scalp and reducing the adverse reactions that occur during the use of minoxidil.

[0016] Secondly, the Usma grass was extracted with deionized water to obtain the Usma grass extract; the Usma grass extract was loaded into human umbilical cord mesenchymal stem cells to obtain stem cell-loaded Usma grass extract; minoxidil-ginsenoside transfer body solution, stem cell-loaded Usma grass extract, and deionized water were mixed to obtain a minoxidil mixture; the Usma grass extract contains an inhibitory component of 5α-reductase, which inhibits the conversion of testosterone into more active DHT and reduces the level of DHT in the blood and hair follicles, thereby alleviating DHT-induced hair follicle miniaturization. Finasteride approved by the U.S. Food and Drug Administration has a similar effect, but finasteride can cause sexual dysfunction and psychological disorders, including erectile dysfunction. Therefore, extracting the inhibitory components of 5α-reductase from natural plants can not only achieve the effect of treating androgenic alopecia, but also greatly reduce the adverse reactions during use; loading the extract of Usma grass with stem cells can not only improve the transdermal properties of the extract of Usma grass, but also secrete various cytokines and growth factors, such as insulin-like growth factor binding protein precursor, platelet-derived growth factor, basic fibroblast growth factor, keratinocyte growth factor, hepatocyte growth factor, vascular endothelial growth factor, etc., which can regulate the hair follicle cycle, promote angiogenesis, anti-apoptosis and promote hair regeneration. DETAILED DESCRIPTION

[0017] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. Embodiment 1:

[0018] A preparation process of a minoxidil mixed solution, the preparation process of the minoxidil mixed solution comprising the following preparation steps: (1) Chloroform and methanol were mixed in a mass ratio of 10:1 to prepare a mixed solvent; soybean lecithin and ginsenoside were added in a mass ratio of 7:2 to a mixed solvent of 4 times the mass of soybean lecithin, and ultrasonically dispersed at room temperature for 10 minutes, and dried at 20°C for 6 hours under vacuum conditions to obtain a phospholipid-ginsenoside film; minoxidil and Tween 80 were added in a mass ratio of 2:0.12 to deionized water of 8 times the mass of minoxidil, and stirred at 10°C and 300 r / min for 20 minutes, and phospholipid-ginsenoside film of 4 times the mass of minoxidil was added, the temperature was raised to 50°C, and the stirring was continued for 2 hours. After cooling to room temperature, the mixture was allowed to stand for 1 hour to obtain a transfersome suspension; the transfersome suspension was ultrasonically dispersed at 0°C for 12 minutes, and filtered through a 0.22µm microporous filter membrane to obtain a minoxidil-ginsenoside transfersome solution; (2) The Usma grass was crushed into Usma grass powder with a particle size of less than 3 mm by a wall breaking machine, the Usma grass powder was mixed with deionized water at a mass ratio of 1:40, stirred at 100°C and 300 r / min for 3 h, filtered with filter cloth, the filter residue and deionized water were mixed at a mass ratio of 1:20, stirred at 100°C and 300 r / min for 1 h, filtered with filter cloth, the two filtrates were mixed, centrifuged at 4500 r / min for 11 min, and the supernatant was dried at 50°C under vacuum conditions for 5 h to obtain the Usma grass extract; human umbilical cord mesenchymal stem cells, trypsin, and P The BS solution was mixed and ultrasonically dispersed at a mass ratio of 1:0.2:80 for 2 minutes, placed in a centrifuge tube, and centrifuged at 900 r / min for 4 minutes. The resulting precipitate was the stem cell membrane; the Usma grass extract, stem cell membrane, and PBS solution were mixed at a mass ratio of 1:3:40, placed in a centrifuge tube, and centrifuged at 900 r / min for 7 minutes. The precipitate was the stem cell-loaded Usma grass extract; the minoxidil-ginsenoside transfersome solution, the stem cell-loaded Usma grass extract, and deionized water were evenly mixed at a mass ratio of 1:0.2:3 to obtain a minoxidil mixed solution. Embodiment 2:

[0019] A preparation process of a minoxidil mixed solution, the preparation process of the minoxidil mixed solution comprising the following preparation steps: (1) Chloroform and methanol were mixed in a mass ratio of 10:1 to prepare a mixed solvent; soybean lecithin and ginsenoside were added in a mass ratio of 7:2 to a mixed solvent of 4.5 times the mass of soybean lecithin, and ultrasonically dispersed at room temperature for 11 minutes, and dried at 25°C for 5.5 hours under vacuum conditions to obtain a phospholipid-ginsenoside film; minoxidil and Tween 80 were added in a mass ratio of 2:0.125 to deionized water of 9 times the mass of minoxidil, stirred at 20°C and 400 r / min for 18 minutes, and phospholipid-ginsenoside film of 4.5 times the mass of minoxidil was added, the temperature was raised to 55°C, and the stirring was continued for 1.5 hours. After cooling to room temperature, the mixture was allowed to stand for 1.5 hours to obtain a transfersome suspension; the transfersome suspension was ultrasonically dispersed at 1°C for 13 minutes, and filtered through a 0.22µm microporous filter membrane to obtain a minoxidil-ginsenoside transfersome solution; (2) The Usma grass was crushed into Usma grass powder with a particle size of less than 3 mm by a wall breaking machine, the Usma grass powder was mixed with deionized water at a mass ratio of 1:45, stirred at 100°C and 400 r / min for 2.5 h, filtered with filter cloth, the filter residue and deionized water were mixed at a mass ratio of 1:22, stirred at 100°C and 400 r / min for 1 h, filtered with filter cloth, the two filtrates were mixed, centrifuged at 4500 r / min for 10 min, and the supernatant was dried at 55°C for 4.5 h under vacuum conditions to obtain the Usma grass extract; human umbilical cord mesenchymal stem cells, trypsin, and PBS solution were added to the mixture. The liquid was mixed in a mass ratio of 1:0.25:90 and ultrasonically dispersed for 2.5 minutes, placed in a centrifuge tube, and centrifuged at 900r / min for 5 minutes. The resulting precipitate was the stem cell membrane; the Usma grass extract, stem cell membrane, and PBS solution were mixed in a mass ratio of 1:3.5:45, placed in a centrifuge tube, and centrifuged at 900r / min for 7.5 minutes. The precipitate was the stem cell-loaded Usma grass extract; the minoxidil-ginsenoside transfersome solution, the stem cell-loaded Usma grass extract, and deionized water were evenly mixed in a mass ratio of 1:0.25:3.5 to obtain a minoxidil mixed solution. Embodiment 3:

[0020] A preparation process of a minoxidil mixed solution, the preparation process of the minoxidil mixed solution comprising the following preparation steps: (1) Chloroform and methanol were mixed in a mass ratio of 10:1 to prepare a mixed solvent; soybean lecithin and ginsenoside were added in a mass ratio of 7:2 to a mixed solvent of 5 times the mass of soybean lecithin, and ultrasonically dispersed at room temperature for 12 minutes, and dried at 30°C for 5 hours under vacuum conditions to obtain a phospholipid-ginsenoside membrane; minoxidil and Tween 80 were added in a mass ratio of 2:0.13 to deionized water of 10 times the mass of minoxidil, and stirred at 30°C and 500 r / min for 16 minutes, and phospholipid-ginsenoside membrane of 5 times the mass of minoxidil was added, the temperature was raised to 60°C, and the stirring was continued for 1 hour. After cooling to room temperature, the mixture was allowed to stand for 2 hours to obtain a transfersome suspension; the transfersome suspension was ultrasonically dispersed at 2°C for 14 minutes, and filtered through a 0.22µm microporous filter membrane to obtain a minoxidil-ginsenoside transfersome solution; (2) The Usma grass was crushed into Usma grass powder with a particle size of less than 3 mm by a wall breaking machine, the Usma grass powder was mixed with deionized water at a mass ratio of 1:50, stirred at 100°C and 500 r / min for 2 h, filtered with filter cloth, the filter residue and deionized water were mixed at a mass ratio of 1:24, stirred at 100°C and 500 r / min for 1 h, filtered with filter cloth, the two filtrates were mixed, centrifuged at 4500 r / min for 11 min, and the supernatant was dried at 60°C under vacuum conditions for 4 h to obtain the Usma grass extract; human umbilical cord mesenchymal stem cells, trypsin, and P The BS solution was mixed at a mass ratio of 1:0.3:100 and ultrasonically dispersed for 3 minutes, placed in a centrifuge tube, and centrifuged at 900 r / min for 6 minutes. The resulting precipitate was the stem cell membrane; the Usma grass extract, stem cell membrane, and PBS solution were mixed at a mass ratio of 1:4:50, placed in a centrifuge tube, and centrifuged at 900 r / min for 8 minutes. The precipitate was the stem cell-loaded Usma grass extract; the minoxidil-ginsenoside transfersome solution, the stem cell-loaded Usma grass extract, and deionized water were evenly mixed at a mass ratio of 1:0.3:4 to obtain a minoxidil mixed solution.

[0021] Comparative Example 1: The preparation process of the minoxidil mixed solution of Comparative Example 1 is different from that of Example 2 in that step (1) is not performed, and step (2) is modified as follows: the Usma grass is crushed into a Usma grass powder with a particle size of less than 3 mm by a wall breaking machine, the Usma grass powder and deionized water are mixed at a mass ratio of 1:45, stirred at 100°C and 400 r / min for 2.5 h, filtered with a filter cloth, the filter residue and deionized water are mixed at a mass ratio of 1:22, stirred at 100°C and 400 r / min for 1 h, filtered with a filter cloth, the two filtrates are mixed, centrifuged at 4500 r / min for 10 min, and the supernatant is dried at 55°C for 4.5 h under vacuum conditions to obtain Usma grass. Sima grass extract; human umbilical cord mesenchymal stem cells, trypsin, and PBS solution are mixed and ultrasonically dispersed at a mass ratio of 1:0.25:90 for 2.5 minutes, placed in a centrifuge tube, and centrifuged at a speed of 900r / min for 5 minutes. The resulting precipitate is a stem cell membrane; Usma grass extract, stem cell membrane, and PBS solution are mixed at a mass ratio of 1:3.5:45, placed in a centrifuge tube, and centrifuged at a speed of 900r / min for 7.5 minutes. The precipitate is a stem cell-carrying Usma grass extract; minoxidil, stem cell-carrying Usma grass extract, and deionized water are mixed at a mass ratio of 1:0.25:3.5 to obtain a minoxidil mixed solution. The remaining steps are the same as in Example 2.

[0022] Comparative Example 2: The preparation process of the minoxidil mixed solution of Comparative Example 2 is different from that of Example 2 in that step (2) is not performed, and step (1) is modified as follows: chloroform and methanol are mixed uniformly at a mass ratio of 10:1 to prepare a mixed solvent; soybean lecithin and ginsenoside are added at a mass ratio of 7:2 to a mixed solvent 4.5 times the mass of soybean lecithin, and ultrasonically dispersed at room temperature for 11 minutes, and dried at 25°C for 5.5 hours under vacuum conditions to prepare a phospholipid-ginsenoside film; minoxidil is added to the mixture; and the mixture is stirred for 1 hour. , Tween 80 was added to deionized water with a mass ratio of 2:0.125 at 9 times the mass of minoxidil, stirred at 20°C and 400r / min for 18min, phospholipid-ginsenoside membrane with a mass of 4.5 times the mass of minoxidil was added, the temperature was raised to 55°C, stirring was continued for 1.5h, cooled to room temperature and allowed to stand for 1.5h to obtain a transfersome suspension; the transfersome suspension was ultrasonicated at 1°C for 13min, filtered through a 0.22µm microporous filter membrane to obtain a minoxidil mixed solution. The remaining steps were the same as in Example 2.

[0023] Comparative Example 3: The preparation process of the minoxidil mixed solution of Comparative Example 3 is different from that of Example 2 only in step (2), and step (2) is modified as follows: the Usma grass is crushed into Usma grass powder with a particle size of less than 3 mm by a wall breaking machine, the Usma grass powder and deionized water are mixed at a mass ratio of 1:45, stirred at 100°C and 400 r / min for 2.5 hours, filtered with a filter cloth, the filter residue and deionized water are mixed at a mass ratio of 1:22, stirred at 100°C and 400 r / min for 1 hour, filtered with a filter cloth, the two filtrates are mixed, centrifuged at 4500 r / min for 10 minutes, and the supernatant is dried at 55°C for 4.5 hours under vacuum conditions to obtain a Usma grass extract; the minoxidil-ginsenoside transfer body solution, the Usma grass extract, and deionized water are mixed at a mass ratio of 1:0.25:3.5 to obtain a minoxidil mixed solution. The remaining steps are the same as those of Example 2.

[0024] Test Example 1 Testing of the efficacy of androgenic alopecia treatment Test method: Thirty-two male mice of similar weight were selected, and an area of ​​3cm×3cm on the back of the mice was marked as the test area. The androgen testosterone propionate was injected subcutaneously into the mice at a dose of 5mg / kg every day for 30 consecutive days, and obvious hair loss in the injected area was observed.

[0025] The 32 male mice were randomly divided into 8 groups, each with 4 mice. They were respectively Example 1-3 groups, Comparative Example 1-3 groups, positive control group, and negative control group; the minoxidil mixed solution prepared in Example 1-3 and Comparative Example 1-3 was applied to the mice in Example 1-3 groups and Comparative Example 1-3 groups, respectively, the commercially available minoxidil solution was applied to the positive control group, and deionized water was applied to the negative control group, 200 μl was applied to each mouse each time, the administration was once a day, and the administration was continued for 15 days, and the hair growth of the test area of ​​the mice was observed to see whether there were adverse reactions such as rash, desquamation, redness and swelling.

[0026] All mice were killed after 15 days, and the skin of the depilated area on the back of the mice was fixed with 4% paraformaldehyde for 24 hours, dehydrated, paraffin-embedded, sliced, stained with hematoxylin for 5 minutes, and rinsed with tap water; the slices were then dehydrated for 5 minutes, stained with eosin dye, dehydrated for 5 minutes, and the stained slices were observed under an ordinary microscope. The morphology of the hair follicles was observed, and the total number of hair follicles in each group was counted, and the average value of each group was taken. The results are shown in Table 1.

[0027] Table 1 Group Whether adverse reactions occur Total number of hair follicles Example 1 Group none 92 Example 2 Group none 93 Example 3 Group none 91 Comparative Example 1 Slight redness and swelling 76 Comparative Example 2 none 75 Comparative Example 3 none 82 Positive control group Obvious redness, swelling, and scaling 59 Negative control group none 34 From the comparison of the experimental data of Example 1 to 3 and Comparative Example 1 to 3, the positive control group and the negative control group in Table 1, it can be found that the minoxidil mixed solution prepared by the present invention has excellent performance in treating androgenic alopecia.

[0028] By comparison, no adverse reactions occurred in the groups of Examples 1 to 3, slight adverse reactions occurred in the group of Comparative Example 1, and obvious adverse reactions occurred in the positive control group, indicating that soybean lecithin and ginsenosides were mixed and dried to obtain a phospholipid-ginsenoside membrane; minoxidil was wrapped with a phospholipid-ginsenoside membrane to obtain a minoxidil-ginsenoside transfersome solution; minoxidil is a drug approved by the U.S. Food and Drug Administration for the treatment of hair loss, which can increase the blood flow from the skin to the scalp by activating potassium channels, provide good blood supply to the hair follicles and promote hair follicle growth. However, minoxidil can cause adverse reactions such as rough hair, itchy scalp, rash, and desquamation; ginsenosides have anti-inflammatory effects and a steroid structure similar to cholesterol, which has a strong direct interaction with phospholipids and can be tightly inserted into the phospholipid bilayer; encapsulating minoxidil in ginsenosides can improve the transdermal absorption of minoxidil, while reducing the stimulation of minoxidil to the scalp and reducing the adverse reactions that occur during the use of minoxidil.

[0029] By comparison, the total number of hair follicles in Example 1-3 groups is greater than the total number of hair follicles in Comparative Example 1-3 groups, the positive control group, and the negative control group, indicating that the soybean lecithin and ginsenoside are mixed and dried to prepare a phospholipid-ginsenoside membrane; minoxidil is wrapped with the phospholipid-ginsenoside membrane to prepare a minoxidil-ginsenoside transfersome solution; minoxidil is a drug approved by the U.S. Food and Drug Administration for the treatment of hair loss, which can increase the blood flow from the skin to the scalp by activating potassium channels, provide good blood supply to the hair follicles and promote hair follicle growth. However, minoxidil can cause adverse reactions such as rough hair, itchy scalp, rash, and desquamation; ginsenosides have anti-inflammatory effects and a steroid structure similar to cholesterol. They have a strong direct interaction with phospholipids and can be tightly inserted into the phospholipid bilayer; encapsulating minoxidil in ginsenosides can improve the transdermal absorption rate of minoxidil; extracting Usma grass with deionized water to obtain Usma grass extract; loading Usma grass extract into human umbilical cord mesenchymal stem cells to obtain stem cell-loaded Usma grass extract; mixing minoxidil-ginsenoside transfer body solution, stem cell-loaded Usma grass extract, and deionized water to obtain minoxidil mixture liquid; Usma grass extract contains 5α-reductase inhibitory components, which inhibit the conversion of testosterone into more active DHT and reduce the DHT levels in the blood and hair follicles, thereby alleviating DHT-induced hair follicle miniaturization; Loading Usma grass extract with stem cells can not only improve the transdermal properties of Usma grass extract, but also secrete various cytokines and growth factors, such as insulin-like growth factor binding protein precursor, platelet-derived growth factor, basic fibroblast growth factor, keratinocyte growth factor, hepatocyte growth factor, vascular endothelial growth factor, etc., which regulate the hair follicle cycle, promote angiogenesis, and resist apoptosis to promote hair regeneration.

[0030] The specific implementation methods described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above description is only a specific implementation method of the present invention and is not intended to limit the scope of protection of the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A process for preparing a minoxidil mixed solution, characterized in that: The preparation process of the minoxidil mixed solution comprises wrapping minoxidil with a phospholipid-ginsenoside membrane to obtain a minoxidil-ginsenoside transfersome solution; loading the Usma grass extract into human umbilical cord mesenchymal stem cells to obtain stem cell-loaded Usma grass extract; and mixing the minoxidil-ginsenoside transfersome solution, the stem cell-loaded Usma grass extract, and deionized water to obtain a minoxidil mixed solution. The phospholipid-ginsenoside membrane is prepared by mixing and drying soybean lecithin and ginsenoside; The Usma grass extract is prepared by extracting Usma grass with deionized water.

2. The process for preparing a minoxidil mixed solution according to claim 1, characterized in that: The preparation process of the minoxidil mixed solution comprises the following preparation steps: (1) Add minoxidil and Tween 80 in a mass ratio of 2:(0.12-0.13) to deionized water 8-10 times the mass of minoxidil, stir at 10-30°C and 300-500 r / min for 16-20 min, add phospholipid-ginsenoside membrane 4-5 times the mass of minoxidil, heat to 50-60°C, continue stirring for 1-2 h, cool to room temperature and let stand for 1-2 h to obtain a transfersome suspension; ultrasonically treat the transfersome suspension at 0-2°C for 12-14 min, filter through a 0.22 µm microporous filter membrane to obtain a minoxidil-ginsenoside transfersome solution; (2) Human umbilical cord mesenchymal stem cells, trypsin and PBS solution were mixed and ultrasonically dispersed for 2-3 minutes, placed in a centrifuge tube, and centrifuged at 900 r / min for 4-6 minutes. The resulting precipitate was the stem cell membrane; Usma grass extract, stem cell membrane and PBS solution were mixed in a mass ratio of 1:(3-4):(40-50), placed in a centrifuge tube, and centrifuged at 900 r / min for 7-8 minutes. The precipitate was the stem cell-loaded Usma grass extract; minoxidil-ginsenoside transfersome solution, stem cell-loaded Usma grass extract and deionized water were evenly mixed to prepare a minoxidil mixed solution.

3. The process for preparing a minoxidil mixed solution according to claim 2, characterized in that: The preparation method of the phospholipid-ginsenoside film in step (1) is as follows: soybean lecithin and ginsenoside are added in a mass ratio of 7:2 to a mixed solvent of 4 to 5 times the mass of soybean lecithin, ultrasonically dispersed at room temperature for 10 to 12 minutes, and dried at 20 to 30° C. for 5 to 6 hours under vacuum conditions to obtain the phospholipid-ginsenoside film.

4. The process for preparing a minoxidil mixed solution according to claim 3, characterized in that: The preparation method of the mixed solvent is: chloroform and methanol are evenly mixed in a mass ratio of 10:1 to prepare a mixed solvent.

5. The process for preparing a minoxidil mixed solution according to claim 2, characterized in that: The preparation method of the Usma grass extract in step (2) is as follows: uniformly mix Usma grass powder and deionized water in a mass ratio of 1: (40-50), stir at 100°C and 300-500 r / min for 2-3 hours, filter through filter cloth, uniformly mix the filter residue and deionized water in a mass ratio of 1: (20-24), stir at 100°C and 300-500 r / min for 1 hour, filter through filter cloth, mix the two filtrates, centrifuge at 4500 r / min for 9-11 minutes, and dry the supernatant at 50-60°C under vacuum conditions for 4-5 hours to obtain the Usma grass extract.

6. The process for preparing a minoxidil mixed solution according to claim 5, characterized in that: The Usma grass powder is prepared by crushing the Usma grass into Usma grass powder with a particle size less than 3 mm using a wall breaking machine.

7. The process for preparing a minoxidil mixed solution according to claim 2, characterized in that: The mass ratio of the human umbilical cord mesenchymal stem cells, trypsin and PBS solution in step (2) is 1:(0.2-0.3):(80-100).

8. The process for preparing a minoxidil mixed solution according to claim 2, characterized in that: The model of the PBS solution in step (2) is CD433577.

9. The process for preparing a minoxidil mixed solution according to claim 2, characterized in that: The mass ratio of the minoxidil-ginsenoside transfersome solution, stem cell-loaded Usma grass extract, and deionized water in step (2) is 1:(0.2-0.3):(3-4).