Preparations for improving female urogenital syndrome and their use

Abstract

This invention provides a formulation for improving female urogenital syndrome and its use. [Solution] A composition is provided for improving genitourinary syndrome in women, which contains an effective amount of growth factors. The effective amount of growth factors is preferably 25 to 120 ng of platelet-derived growth factor BB (PDGF-BB).

Description

Technical Field

[0001] The present invention relates to a preparation for improving female urogenital syndrome and its use, and particularly relates to a preparation containing an effective amount of platelet-derived growth factor (PDGF), and the use of the preparation for improving female urogenital syndrome.

Background Art

[0002] Generally, the defensive structure against external stimuli in the outermost layer of the skin is the stratum corneum, which usually consists of multiple layers of dead flattened cells densely overlapping. If this is too much or too thick, so-called "keratoderma" is formed, and the skin secretes sebum to form a sebum film with a waterproof function on the surface layer of the stratum corneum. However, the skin in the delicate zone (primary sexual characteristic site of women) of women is different from general skin. The mucosal-like skin of the vulva and urethral orifice has only a basal layer, a parabasal layer, and a stratum corneum, is very soft and elastic, but not as tough as general skin.

[0003] The stratum corneum of mucosal-like skin does not have a sebum film and is immersed in mucus, so waterproofing is impossible. In addition, the stratum corneum of mucosal-like skin has a thin and soft structure and lacks lipids, so moisture is easily flowing. Such a structure contributes to the passage of lactic acid bacteria, immune system cells and molecules, and forms a very special microecosystem.

[0004] Genitourinary syndrome, also known as reproductive urinary syndrome, is a condition that affects women. It is caused by factors such as pressure, irregular lifestyle, unbalanced diet, clothing habits, and menopause, leading to atrophy and dryness of epithelial cells in the urinary tract or genitals, breakdown of subcutaneous connective tissue, loss of collagen, and decreased hyaluronic acid production. This results in symptoms such as itching and pain due to dryness of the urinary tract or genitals, urinary incontinence, frequent urination, and discomfort during sexual intercourse. Current treatments include hormone replacement therapy and topical medications, but both have side effects. In addition, current treatments to improve genitourinary syndrome involve injections of hyaluronic acid or autologous fat, which are injected into multiple sites. For example, the O-Shot technique, advocated by Dr. Charles Lunels in the United States, involves injecting platelet-rich plasma (PRP) in a circular pattern into the soft tissue of the vagina. However, since the soft tissue in question may include at least the vaginal opening, anterior vaginal wall, posterior vaginal wall, G-spot, anus, and urethra, it can not only cause discomfort in a woman's delicate areas, but excessive stimulation may also trigger allergies, swelling, and inflammation in these areas, and excessive moisture may lead to fungal infections.

[0005] Therefore, the inventor aims to provide a formulation that improves female urogenital syndrome, thereby enabling the safe remission and improvement of symptoms of female urogenital syndrome. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] The present invention provides a formulation containing a growth factor (Platelet-derived growth factor, PDGF) for the improvement of female urogenital syndrome. [Means for solving the problem]

[0007] The composition of the present invention is used to improve female urogenital syndrome. The composition is a formulation containing a growth factor (platelet-derived growth factor, PDGF). The composition includes, but is not limited to, a dried powder formulation, a frozen formulation, or a re-dissolving formulation.

[0008] The present invention relates to the use of a composition for preparing a formulation to improve female urogenital syndrome, wherein the composition contains an effective amount of growth factors. The composition includes, but is not limited to, a dried powder formulation, a frozen formulation, or a re-dissolving formulation.

[0009] Preferably, the effective amount of the growth factor (Platelet-derived growth factor, PDGF) is 25 to 120 ng.

[0010] Preferably, the growth factor is platelet-derived growth factor (PDGF), specifically platelet-derived growth factor AA (PDGF-AA), platelet-derived growth factor AB (PDGF-AB), or platelet-derived growth factor BB (PDGF-BB). More preferably, the growth factor is platelet-derived growth factor BB (PDGF-BB).

[0011] The formulation of the present invention is a parenteral formulation. Preferably, the formulation of the present invention can be mixed with a solvent to make it injectable, or prepared as a topical formulation such as a cream, ointment, gel, or suppository.

[0012] Preferably, the formulation of the present invention is administered by infusion.

[0013] In this invention, the term "delicate zone" refers to the area of ​​a woman's primary sexual characteristics, and includes, but is not limited to, the external and internal genitalia. Based on an anatomical definition, the internal genitalia include the vagina, uterus, fallopian tubes, and ovaries, while the external genitalia include the mons pubis, labia majora, labia minora, clitoris, vaginal opening, vaginal vestibule, Bartholin's glands, hymen, and perineum, etc.

[0014] In a preferred embodiment of the present invention, the formulation is injected into two areas of the delicate zone. In a more preferred embodiment of the present invention, the formulation is injected into the anterior vaginal wall and the clitoris.

[0015] The genitourinary syndrome described in this invention is caused by pressure, an irregular lifestyle, an unbalanced diet, clothing habits, or menopause. Preferably, the genitourinary syndrome described in this invention is genital urinary syndrome of menopause.

[0016] In the present invention, the platelet-derived growth factor is obtained from mammalian platelets. Preferably, the platelet-derived growth factor is obtained from human or porcine platelets, and more preferably, the platelet-derived growth factor is obtained from human platelets.

[0017] In this text, the term "improve" means that when a patient uses the formulation of the present invention, the course of the disease or symptoms are relieved or eliminated.

[0018] The formulation of the present invention can be used in combination with a pharmaceutically acceptable solvent. Such pharmaceutically acceptable solvents include, for example, saline solution, water for injection, hyaluronic acid, lidocaine, or steroids, which are solvents capable of dissolving human or veterinary medicinal products. The amount of solvent added should, in principle, be such that an effective concentration of the growth factor is maintained.

[0019] The formulations of the present invention may contain pharmaceutically acceptable excipients, and it is necessary to use well-known excipients depending on the dosage form. [Brief explanation of the drawing]

[0020] [Figure 1] Figure 1 shows the content of platelet-derived growth factor BB (PDGF-BB) in the dried platelet powder prepared according to the present invention. [Figure 2] Figure 2 shows the test process of the present invention. [Figure 3] Figure 3 is a schematic diagram of the locations where collagen density (intensity) and mucosal layer thickness (skin thickness) were recorded in the present invention. [Figure 4] Figure 4 shows images and numerical values ​​of collagen density (intensity) and mucosal layer thickness (skin thickness) for five subjects before administration of the formulation of the present invention (hereinafter referred to as "formulation administration"), 4 weeks after the first formulation administration, and 4 weeks after the second formulation administration. [Figure 5] Figure 5 is a bar graph showing the skin thickness of the mucosal layer of five subjects before administration of the drug, 4 weeks after the first administration of the drug, and 4 weeks after the second administration of the drug. [Figure 6] Figure 6 is a bar graph showing the collagen density (intensity) of five subjects before administration of the formulation, 4 weeks after the first administration of the formulation, and 4 weeks after the second administration of the formulation. [Figure 7] Figure 7 shows images and numerical values ​​of collagen density (intensity) and mucosal layer thickness (skin thickness) for subject No. 4 before administration of the formulation, 4 weeks after the first administration of the formulation, 4 weeks after the second administration of the formulation, 8 weeks after the second administration of the formulation, and 12 weeks after the second administration of the formulation. [Figure 8] Figure 8 is a bar graph showing the skin thickness and collagen intensity of subject No. 4 before administration of the drug, 4 weeks after the first administration, 4 weeks after the second administration, 8 weeks after the second administration, and 12 weeks after the second administration.

Mode for Carrying Out the Invention

[0021] It should be understood that the detailed description of the embodiments is for explaining the preferred embodiments of the present invention and is not intended to limit the present invention to any of the embodiments. It should be noted that the present invention is intended to cover all alternative embodiments within the same spirit and scope of the present invention.

[0022] Experiment 1: Preparation of platelet dry powder

[0023] In the present invention, platelet dry powder can be prepared from porcine whole blood or human whole blood.

[0024] In an embodiment of the present invention, human whole blood is put into a centrifuge tube containing an anticoagulant, and the human whole blood is centrifuged using a centrifuge (1500 - 2000 × g, 10 - 15 minutes. This is referred to as primary centrifugation), so that the human whole blood is separated into three layers in order from top to bottom: a plasma layer, a buffy coat layer where white blood cells are present, and a red blood cell layer. These steps and the layer separation situation of blood cells after centrifugation are well known to those skilled in the art. And the centrifugation conditions of the primary centrifugation can be extended to 500 - 3000 × g, 5 - 20 minutes. The platelets after centrifugation were in the plasma layer and were distributed at a position close to the buffy coat layer. Therefore, by sucking the plasma layer and the buffy coat layer in a sterile operation method, a mixture of the plasma layer and the buffy coat layer was obtained. This is platelet-rich plasma (PRP).

[0025] Platelet-rich plasma (PRP) was centrifuged using a centrifuge (700-1000 × g, 5 minutes). The centrifugal conditions can be expanded to 100-1200 × g, 5-10 minutes. Furthermore, the centrifugation speed must be lower than the actual centrifugation speed used in the primary centrifugation step (500-3000 g, 5-20 minutes). This separated the PRP into two layers. Next, the upper and lower layers were separated using aseptic techniques, completely removing leukocytes (WBCs) from the lower layer. Additionally, the number of platelets in the upper layer was detected using a fully automated hematology analyzer (model: XP-300, manufacturer: Sysmex), and the platelet concentration of the upper layer was determined to be 1 × 10⁻⁶. 9 The solution was adjusted to / mL. Then, to avoid a decrease in the concentration of growth factors due to platelet damage, dysfunction, or decreased activity, reagents were added. The reagents may be, for example, calcium chloride, thrombin, or adenosine diphosphate. Subsequently, the solution was centrifuged at 2500-3000 × g for 10 minutes. Note that the centrifugation conditions can be expanded to 500-4000 × g for 10-15 minutes. Furthermore, the rotation speed must be higher than the rotation speed used in the actual centrifugation step (500-3000 g, 5-20 minutes). This concentrated the platelets at the bottom and completely removed the supernatant, thereby completely removing plasma and proteins in the plasma. Then, by adding an equal volume of water for injection to the supernatant, a solution for dispensing was obtained.

[0026] Dispensing solution was dispensed into sample vials. Each sample vial was then filled with 1 ml (mL) of dispensing solution, and then freeze-dried. The freeze-drying steps were as follows: (1) pre-cooling (-30 to -50°C for at least 5 hours), (2) primary drying (gradually raising the temperature from the temperature in step (1) to 10°C for 10 to 20 hours, with a vacuum of 100 mTorr or less), and (3) secondary drying (20°C for 3 to 5 hours, with a vacuum of 100 mTorr or less). The dried material after freeze-drying was platelet dry powder.

[0027] Platelet dry powder was mixed with 1 cc of sterile water for injection to obtain a platelet dry powder reconstitution solution. Next, the platelet-derived growth factor BB (PDGF-BB) content in the platelet dry powder reconstitution solution was analyzed using an enzyme-linked immunosorbent assay (ELISA).

[0028] The experimental results are shown in Table 1 and Figure 1. The PDGF-BB content in each vial of platelet-dried powder prepared according to the present invention was 10 ng / mL or higher.

[0029] [Table 1]

[0030] Experiment 2: Efficacy study for female genitourinary syndrome

[0031] In this invention, we recruited female volunteers suffering from urogenital syndrome. The volunteers were between 20 and 50 years old.

[0032] First, volunteers were given the Female Sexual Function Index (FSFI) questionnaire to complete. The questionnaire consisted of 19 items on themes related to symptoms of genitourinary syndrome, including dryness of the genital area and discomfort during sexual intercourse. Each theme had six options. The six options corresponded to 1, 2, 3, 4, and 5 points respectively, in descending order of degree or frequency of being asked about the theme, and a score of 0 was recorded if the behavior related to the theme was not performed.

[0033] The results of the FSFI questionnaire are largely based on subjective feelings. Furthermore, during the experiment, some subjects did not engage in sexual activity due to menstruation or other factors, so the FSFI questionnaire results were not desirable. Therefore, in this invention, further scientific detection was performed.

[0034] Based on the results of the questionnaire, suitable volunteers to participate in the study were selected and referred to as subjects. The study process was as shown in Figure 2. First, on day 0, each subject underwent detection using a collagen detection device (DermaLab® Series SkinLab USB) before being administered the formulation.

[0035] After filling the probe with pure water and sealing it with plastic wrap, it was connected to the main body of the collagen detection device. Next, conductive gel was applied to the probe and it was lightly pressed against the mucous membrane in the lower left corner of the vaginal opening (indicated by the arrow in Figure 3). Then, the capture button was pressed to take an ultrasound image of the mucosal layer, and the collagen density (intensity) and skin thickness (skin thickness) were recorded.

[0036] Platelet-free powder was redissolved in 2% lidocaine, and the total amount of growth factor PDGF-BB was calculated to be 25-120 ng. This was then inoculated into two sites of the subject: the anterior vaginal wall and the clitoris. The total inoculation volume was 1-7 mL. In a preferred embodiment, the total inoculation volume was 5 mL. Furthermore, the inoculation volume to the anterior vaginal wall was greater than or equal to the inoculation volume to the clitoris. The ratio of inoculation volumes to the anterior vaginal wall and the clitoris was 80-60% and 40-20%, respectively. In the most preferred embodiment, the inoculation volume to the anterior vaginal wall was 3-4 mL, and the inoculation volume to the clitoris was 1-2 mL.

[0037] Four weeks after administering the formulation to the subjects, the second drug was administered, and the completion of the index questionnaire and detection using the collagen detection device were repeated. In addition, to ensure the accuracy of the study, in accordance with the natural metabolism caused by the women's menstrual cycle, the completion of the index questionnaire and detection using the collagen detection device were repeated at weeks 4, 8, and 12 after the administration of the second drug.

[0038] Finally, the results of five subjects were randomly selected and analyzed.

[0039] The collagen density (intensity) and skin thickness (skin thickness) results for five subjects before administration of the formulation, 4 weeks after administration of the first drug, 4 weeks after administration of the second drug, 8 weeks after administration of the second drug, and 12 weeks after administration of the second drug are shown in Table 2, Figures 4, 5, and 6.

[0040] [Table 2]

[0041] As is evident from the results regarding skin thickness, with the exception of subject number 1, who did not show any significant improvement in skin thickness four weeks after administration of the first drug, all other subjects showed a remarkable improvement in skin thickness four weeks after administration of the first drug.

[0042] Due to individual differences, the thickness of the mucosal layer (Skin Thickness) of subjects 4 weeks after administration of the second drug showed different results compared to the thickness of the mucosal layer (Skin Thickness) 4 weeks after administration of the first drug, including a decrease in thickness (No. 1, No. 2), maintenance (No. 5), or improvement (No. 3, No. 4). However, compared to the condition of each subject before administration of the formulation, all subjects except for subject No. 1 showed a significant improvement in the thickness of the mucosal layer (Skin Thickness).

[0043] The subjects were followed up for 12 weeks after the administration of the second drug and compared to their condition before drug administration. In subject #1, although no significant improvement in mucosal layer thickness (skin thickness) was observed, the thickness remained the same as before drug administration, and no significant decrease in thickness was observed. In addition, all other subjects showed a significant improvement in mucosal layer thickness (skin thickness). Among them, subject #4 showed the greatest improvement in mucosal layer thickness (skin thickness).

[0044] As is evident from the collagen density (intensity) results, with the exception of subjects #2 and #3, who showed a slight decrease in collagen density (intensity) four weeks after administration of the first drug, all other subjects showed a significant improvement in collagen density (intensity) four weeks after administration of the first drug.

[0045] Due to individual differences, the collagen density (intensity) of subjects 4 weeks after administration of the second drug showed different results compared to the collagen density (intensity) 4 weeks after administration of the first drug, such as a decrease in density (Nos. 2, 4, and 5) or an improvement (Nos. 1 and 3). However, compared to the condition of each subject before administration of the formulation, with the exception of subject No. 2, all other subjects, including subject No. 3 who showed a slight decrease in collagen density (intensity) 4 weeks after administration of the first drug, showed a significant improvement in collagen density (intensity).

[0046] Follow-up was continued for 12 weeks after the administration of the second drug, and compared with the condition of each subject before administration of the drug. In subject number 2, collagen density (intensity) was clearly lower than the condition before administration of the drug, but skin thickness was significantly improved, so it was determined that the drug had an effect on improving urogenital syndrome. In addition, all other subjects showed a significant improvement in collagen density (intensity). Among them, subjects number 4 and 5 showed the greatest improvement in collagen density (intensity).

[0047] The treatment status and trends of subject number 4 are shown in Figures 7 and 8. As is clear from the figures, subject number 4 showed a tendency for collagen density (intensity) and mucosal layer thickness (skin thickness) to improve and then decrease as the time after administration of the drug increased during the treatment process. However, compared to before administration of the drug, the symptoms of genitourinary syndrome were clearly improved.

[0048] Although the present invention is described and illustrated in sufficient detail so that those skilled in the art can manufacture and implement it, they may also conceive of modifications and other uses. Such modifications are in line with the spirit of the invention and are defined by the claims.

Claims

1. A composition for improving female genitourinary syndrome, comprising an effective amount of growth factors.

2. The composition according to claim 1, wherein the effective amount of growth factor is 25 to 120 ng of platelet-derived growth factor-BB (PDGF-BB).

3. The platelet-derived growth factor-BB is obtained from mammalian platelets, according to claim 2.

4. The aforementioned composition, The first step involves centrifuging whole blood to obtain platelet-rich plasma. The platelet-rich plasma is subjected to a second-stage centrifugation to obtain two separate layers, upper and lower. The upper separation layer is subjected to a third-stage centrifugation to remove the supernatant and obtain platelet precipitate, and The steps include adding a solvent and mixing it with the platelet precipitate, and then performing freeze-drying to obtain the composition, The composition according to claim 1, which is prepared by a method comprising:

5. The composition according to claim 1, wherein the genitourinary syndrome includes genital urinary syndrome of menopause.

6. Use of a composition for preparing a formulation to improve female urogenital syndrome, wherein the composition contains an effective amount of growth factors.

7. The use according to claim 6, wherein the effective amount of growth factors is 25 to 120 ng of platelet-derived growth factor BB (PDGF-BB).

8. The use of the platelet-derived growth factor-BB according to claim 6, obtained from mammalian platelets.

9. The use of the preparation according to claim 6, wherein the preparation is injected by injection into any two sites of primary sexual characteristics.

10. The use of the preparation according to claim 9, wherein the preparation is injected only into the anterior vaginal wall and clitoris.

11. The use according to claim 10, wherein the total volume of the injection is 1 to 7 mL, and the volume injected into the anterior vaginal wall is greater than or equal to the volume injected into the clitoris.

12. The use according to claim 10, wherein the ratio of the volume injected into the anterior vaginal wall is 80-60%, and the ratio of the volume injected into the clitoris is 40-20%.

13. The use according to claim 6, wherein the genitourinary syndrome includes genital urinary syndrome of menopause.

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