Application of urofollitropin for injection combined with letrozole in the preparation of a kit for infertility

Abstract

The present invention relates to the technical field of infertility treatment, and particularly to the application of urofollitropin for injection combined with letrozole in the preparation of a kit for infertility. The present invention for the first time combines urofollitropin for injection and letrozole to prepare soluble microneedles. Specifically, urofollitropin for injection and letrozole are used as drug active ingredients, and the drug active ingredients are loaded in the form of PLGA-chitosan cross-linked microspheres to prepare soluble microneedles. Through verification and analysis, it is found that the soluble microneedles can effectively solve the problem of medication compliance existing in the prior art and can greatly improve the technical effect of traditional treatment methods.

Description

Technical Field

[0001] The present invention relates to the technical field of infertility treatment, and particularly relates to the application of urofollitropin for injection combined with letrozole in the preparation of a kit for infertility treatment. Background Art

[0002] The medical definition of infertility is that a couple has not taken any contraceptive measures for more than one year, has normal sexual life but has not achieved a successful pregnancy. It is mainly divided into primary infertility and secondary infertility. Primary infertility means never having been pregnant; secondary infertility means having been pregnant but then becoming infertile. According to this strict definition, infertility is a common problem that affects at least about 10% - 15% of reproductive-age couples. The causes of infertility are divided into male infertility and female infertility.

[0003] Among them, the causes of infertility in women account for about 45%. ① Affecting egg development: Disorders of the hypothalamic-pituitary-ovarian axis can affect egg maturation or interfere with egg ovulation; congenital ovarian hypoplasia, surgical removal of the ovaries, or changes in the external environment and mental and emotional impacts can all affect ovulation. ② Affecting the transportation of eggs, sperm, or fertilized eggs: After the eggs are discharged, they enter the fallopian tubes. After the sperm are ejaculated into the vagina, they also need to pass through the uterine cavity and fallopian tubes to meet the eggs, and the fertilized eggs also enter the uterine cavity through the fallopian tubes. Studies have found that there is a mycoplasma in the vaginal, cervical secretions, and endometrium of many infertile women with reproductive tract inflammation, which may be the cause of infertility. It is estimated that it is related to the mycoplasma attaching to the sperm tail and affecting sperm motility. Among infertile women with unknown causes, the detection rate of mycoplasma is as high as 85 - 95%, while the control group of fertile women is 23%. After treatment, the pregnancy rate can be significantly increased. ③ Affecting sperm ascent: Such as vaginal septum, vaginal and cervical inflammation, cervical os atresia, or endocrine disorders resulting in overly thick cervical mucus, etc.

[0004] Clinically, urofollitropin for injection is usually used to promote follicle maturation and estrogen secretion, which is suitable for anovulatory symptoms. However, since the treatment requires multiple strict dosages to be injected to avoid overstimulation of the ovaries by ovulation-stimulating drugs, repeated injections will cause pain and discomfort to the patient, and may also cause adverse reactions such as local redness, swelling, and induration due to allergies, bringing great pain and pressure to the patient physically and mentally. In addition, the prior art also discloses the effect of using urofollitropin for injection combined with letrozole to assist in the treatment of infertility patients by intrauterine insemination. However, this treatment method still fails to effectively overcome the physical and psychological pressures of the subjects during the treatment process, and at the same time, its treatment effect also needs to be further improved. Summary of the Invention

[0005] In view of the technical problems existing in the prior art, the present invention provides the use of urofollitropin for injection in combination with letrozole in the preparation of a kit for infertility. In the kit of the present invention, specifically, urofollitropin for injection and letrozole are used as active pharmaceutical ingredients, and the active pharmaceutical ingredients are loaded in the form of PLGA-chitosan cross-linked microspheres to prepare soluble microneedles. Through verification and analysis, the soluble microneedles can effectively solve the problem of medication compliance in the prior art and can greatly improve the technical effect of traditional treatment methods.

[0006] One of the purposes of the present invention is to provide a method for preparing soluble microneedles of urofollitropin for injection in combination with letrozole, which is characterized in that the method comprises the following steps:

[0007] 1) Dissolve 100 - 200 mg of poly (lactic-co-glycolic acid) copolymer in 1 - 2 mL of dichloromethane to obtain a mixed solution 1 for standby;

[0008] 2) Dissolve 1 - 2 g of chitosan in 0.1 M glacial acetic acid and adjust the pH of the solution to 7.0 with NaOH to obtain a mixed solution 2 for standby;

[0009] 3) Dissolve 2.5 - 7.5 mg of letrozole tablets in 2 - 6 mL of 75 IU urofollitropin for injection to obtain a mixed solution 3 for standby;

[0010] 4) Add the mixed solution 3 obtained in step 3) to the mixed solution 2, and after mixing evenly, obtain a mixed solution 4 for standby;

[0011] 5) Add the mixed solution 4 obtained in step 4) to the mixed solution 1 in step 1), and simultaneously add 500 - 1000 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-cross-linked nanomicroparticles for standby;

[0012] 6) Take 1 mL of the chitosan-cross-linked nanomicroparticle solution obtained in step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.), place the mold in a centrifuge, and centrifuge at 4000 - 5000 r / min at 4 °C for 5 - 10 min to make the solution uniformly fill the micropores of the mold. Then place the mold in a freeze dryer, and after drying, obtain soluble microneedles of urofollitropin for injection in combination with letrozole.

[0013] Preferably, the method for preparing soluble microneedles of urofollitropin for injection in combination with letrozole comprises the following steps:

[0014] 1) Dissolve 100 mg of poly (lactic-co-glycolic acid) copolymer (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 1 mL of dichloromethane to obtain a mixed solution 1 for standby;

[0015] 2) Dissolve 1 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby.

[0016] 3) Dissolve 2.5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 2 mL of 75 IU urofollitropin for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group) to obtain mixture 3 for standby.

[0017] 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2). After mixing evenly, obtain mixture 4 for standby.

[0018] 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 500 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby.

[0019] 6) Take 1 mL of the chitosan-crosslinked nanosphere solution in step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.). Place the mold in a centrifuge and centrifuge at 5000 r / min at 4 °C for 5 min to make the solution uniformly fill the micropores of the mold. Then place the molds in a freeze dryer. After drying, obtain soluble microneedles of urofollitropin for injection combined with letrozole.

[0020] Preferably, the preparation method of the soluble microneedles of urofollitropin for injection combined with letrozole includes the following steps:

[0021] 1) Dissolve 150 mg of poly (lactic-co-glycolic acid) (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 1.5 mL of dichloromethane to obtain mixture 1 for standby.

[0022] 2) Dissolve 1.5 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby.

[0023] 3) Dissolve 5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 4 mL of 75 IU urofollitropin for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group) to obtain mixture 3 for standby.

[0024] 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2). After mixing evenly, obtain mixture 4 for standby.

[0025] 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 800 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby.

[0026] 6) Take 1 mL of the chitosan-crosslinked nanospheres solution from step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.). Place the mold in a centrifuge and centrifuge at 5000 r / min at 4 °C for 5 min to uniformly fill the micropores of the mold with the solution. Then place the molds in a freeze dryer, and after drying, obtain the soluble microneedles of urofollitropin for injection combined with letrozole.

[0027] Preferably, the preparation method of the soluble microneedles of urofollitropin for injection combined with letrozole comprises the following steps:

[0028] The preparation method of the soluble microneedles of urofollitropin for injection combined with letrozole is characterized in that the method comprises the following steps:

[0029] 1) Dissolve 200 mg of poly (lactic-co-glycolic acid) (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 2 mL of dichloromethane to obtain mixture 1 for standby;

[0030] 2) Dissolve 2 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby;

[0031] 3) Dissolve 7.5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 6 mL of 75 IU urofollitropin for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group), to obtain mixture 3 for standby;

[0032] 4) Add the mixture 3 obtained in step 3) to the mixture 2, and after mixing evenly, obtain mixture 4 for standby;

[0033] 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 1000 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby;

[0034] 6) Take 1 mL of the chitosan-crosslinked nanospheres solution from step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.). Place the mold in a centrifuge and centrifuge at 5000 r / min at 4 °C for 5 min to uniformly fill the micropores of the mold with the solution. Then place the molds in a freeze dryer, and after drying, obtain the soluble microneedles of urofollitropin for injection combined with letrozole.

[0035] Furthermore, an aspect of the present invention is to provide a soluble microneedle of urofollitropin for injection combined with letrozole, which is characterized in that the soluble microneedle uses PLGA-chitosan crosslinked microspheres as the microneedle body and internally loads urofollitropin for injection and letrozole as active ingredients.

[0036] Preferably, the soluble microneedles are prepared by the above method.

[0037] More preferably, the length of the tip of the soluble microneedles of the present invention is between 100 μm and 1 mm.

[0038] More preferably, the tip of the microneedle can also be embedded with triptorelin acetate for injection, recombinant human follicle-stimulating hormone for injection, menotropins for injection, cetrorelix acetate for injection, etc.

[0039] An aspect of the present invention is to provide the use of a soluble microneedle of urofollitropin for injection combined with letrozole in the preparation of a kit for treating infertility, characterized in that the kit contains the soluble microneedle.

[0040] More preferably, the administration method of the soluble microneedle is to directly attach the microneedle to the lower abdomen once a night for 5 consecutive days.

[0041] The advantages of the present invention are as follows: The present invention combines urofollitropin for injection and letrozole for the first time to prepare soluble microneedles. Specifically, urofollitropin for injection and letrozole are used as drug active ingredients, and the drug active ingredients are loaded in the form of PLGA-chitosan cross-linked microspheres to prepare soluble microneedles. Through verification and analysis, the soluble microneedles can effectively solve the problem of drug compliance in the prior art and can greatly improve the technical effect of traditional treatment methods. Specific Embodiments

[0042] The following specific embodiments are used to further illustrate the present invention in detail, so that those skilled in the art can understand the present invention more clearly.

[0043] The following examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. Based on the specific embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative work belong to the protection scope of the present invention.

[0044] In the embodiments of the present invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art; in the embodiments of the present invention, unless specifically specified, the technical means used are all conventional means well known to those skilled in the art.

[0045] Example 1

[0046] A preparation method of a soluble microneedle of urofollitropin for injection combined with letrozole, characterized in that the method comprises the following steps:

[0047] 1) Dissolve 100 mg of poly (lactic-co-glycolic acid) copolymer (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 1 mL of dichloromethane to obtain mixture 1 for standby;

[0048] 2) Dissolve 1 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby;

[0049] 3) Dissolve 2.5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 2 mL of 75 IU urinary follicle-stimulating hormone for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group) to obtain mixture 3 for standby;

[0050] 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby;

[0051] 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 500 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby;

[0052] 6) Take 1 mL of the chitosan-crosslinked nanosphere solution in step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.), place the mold in a centrifuge, and centrifuge at 5000 r / min at 4 °C for 5 min to make the solution uniformly fill the micropores of the mold. Then place the molds in a freeze dryer, and after drying, obtain soluble microneedles of urinary follicle-stimulating hormone combined with letrozole for injection.

[0053] Example 2

[0054] A preparation method of nanospheres of urinary follicle-stimulating hormone combined with letrozole for injection, characterized in that the method comprises the following steps:

[0055] 1) Dissolve 150 mg of poly (lactic-co-glycolic acid) (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 1.5 mL of dichloromethane to obtain mixture 1 for standby;

[0056] 2) Dissolve 1.5 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby;

[0057] 3) Dissolve 5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 4 mL of 75 IU urinary follicle-stimulating hormone for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group) to obtain mixture 3 for standby;

[0058] 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby;

[0059] 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 800 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby;

[0060] 6) Take 1 mL of the chitosan-crosslinked nanospheres solution from step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.). Place the mold in a centrifuge and centrifuge at 5000 r / min at 4 °C for 5 min to uniformly fill the micropores of the mold with the solution. Then place the molds in a freeze dryer and obtain the soluble microneedles of follicle-stimulating hormone combined with letrozole for injection after drying.

[0061] Example 3

[0062] A preparation method of soluble microneedles of follicle-stimulating hormone combined with letrozole for injection, characterized in that the method comprises the following steps:

[0063] 1) Dissolve 200 mg of poly (lactic-co-glycolic acid) (purchased from Shanghai Yuanye Bio-Technology Co., Ltd.) in 2 mL of dichloromethane to obtain a mixed solution 1 for standby;

[0064] 2) Dissolve 2 g of chitosan in 0.1 M glacial acetic acid and adjust the pH of the solution to 7.0 with NaOH to obtain a mixed solution 2 for standby;

[0065] 3) Dissolve 7.5 mg of letrozole tablets (purchased from Jiangsu Hengrui Medicine Co., Ltd.) in 6 mL of 75 IU follicle-stimulating hormone for injection (purchased from Livzon Pharmaceutical Factory, Livzon Group) to obtain a mixed solution 3 for standby;

[0066] 4) Add the mixed solution 3 obtained in step 3) to the mixed solution 2, and after mixing evenly, obtain a mixed solution 4 for standby;

[0067] 5) Add the mixed solution 4 obtained in step 4) to the mixed solution 1 in step 1), and simultaneously add 1000 mg of tripolyphosphate for crosslinking reaction to obtain chitosan-crosslinked nanospheres for standby;

[0068] 6) Take 1 mL of the chitosan-crosslinked nanospheres solution from step 5) and add it to a microneedle mold (purchased from Suzhou Zhongxin Qiheng Scientific Instruments Co., Ltd.). Place the mold in a centrifuge and centrifuge at 5000 r / min at 4 °C for 5 min to uniformly fill the micropores of the mold with the solution. Then place the molds in a freeze dryer and obtain the soluble microneedles of follicle-stimulating hormone combined with letrozole for injection after drying.

[0069] Control group 1

[0070] The inventors prepared microneedles containing only the same dose of letrozole in the same batch as control 1, and the preparation method was the same as that in Example 1.

[0071] Control group 2

[0072] The inventors prepared only micro - needles containing the same dosage of follicle - stimulating hormone (FSH) in the same batch as Control 2, and the preparation method was the same as that in Example 1.

[0073] Control Group 3

[0074] The inventors used the conventional combination of letrozole and follicle - stimulating hormone as Control Group 3, and the specific administration procedure was as follows: Starting from the menstrual period of the subjects, letrozole 2.5 mg was orally administered once a day for 5 consecutive days, and then 75 IU of human menopausal gonadotropin was intramuscularly injected once a day for 5 consecutive days.

[0075] Verification Example 1

[0076] To accurately evaluate the efficacy of the soluble micro - needles described in the present invention, the inventors selected 60 infertile patients under 40 years old based on clinical examination reports as treatment subjects, grouped them into groups of 10, and respectively administered the soluble micro - needles of injectable follicle - stimulating hormone combined with letrozole prepared in Control Groups 1 - 3 and Example 1 of the present invention. Among them, the administration method of the micro - needles was to directly attach the micro - needles to the lower abdomen once every night for 5 consecutive days. Then, the endometrial thickness, the number of ovulations, and the release amount of sex hormones during the follicular phase in each treatment group were statistically analyzed and compared.

[0077] The results are shown in Table 1 below: Compared with Control Groups 1 - 2, the soluble micro - needles of the present invention can effectively increase the endometrial thickness, and the difference is extremely significant (p < 0.005). At the same time, under the treatment with the micro - needles of the present invention, the number of mature follicles in the subjects is also significantly improved, which can meet the normal fertilization requirements and provide a theoretical basis for the treatment of infertility.

[0078] Table 1 Analysis of endometrial thickness and number of mature follicles in each treatment group

[0079]

[0080]

[0081] In addition, in addition to the secretion of follicular sex hormones being crucial for promoting ovulation, stimulating the development and maturation of follicles is also very important. The inventors continued to monitor the release levels of sex hormones in the subjects in different treatment groups, and the results are shown in Table 2. In the micro - needle treatment groups of Examples 1 - 3, the contents of FSH, LH, and E2 in the patients' bodies were significantly improved, and the difference was statistically significant (P < 0.005).

[0082] Table 2 Analysis of sex hormone secretion in each treatment group

[0083]

[0084] The above results indicate that the soluble microneedles of menotropins for injection combined with letrozole prepared by the present invention have unique advantages in promoting follicle maturation, increasing sex hormone levels, and increasing endometrial thickness in subjects. Moreover, compared with the traditional treatment method of menotropins for injection combined with letrozole, the use of the soluble microneedle method of the present invention can not only meet the normal treatment needs, but also effectively reduce the drug administration burden of the subjects, effectively relieve the resistance of the subjects to the conventional oral and injection methods, and improve the drug administration compliance of the subjects.

[0085] The role of the above embodiments is to illustrate the substantial content of the present invention, but does not limit the protection scope of the present invention. Those of ordinary skill in the art should understand that the technical solutions of the present invention can be modified or equivalently replaced without departing from the essence and protection scope of the technical solutions of the present invention.

Claims

1. A preparation method of a soluble microneedle of urinary follicle-stimulating hormone for injection combined with letrozole, characterized in that, The method includes the following steps: 1) Dissolve 100 - 200 mg of poly (lactic - co - glycolic acid) in 1 - 2 mL of dichloromethane to obtain mixture 1 for standby; 2) Dissolve 1 - 2 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby; 3) Dissolve 2.5 - 7.5 mg of letrozole tablets in 2 - 6 mL of 75 IU urinary follicle - stimulating hormone for injection to obtain mixture 3 for standby; 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby; 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 500 - 1000 mg of tripolyphosphate for cross - linking reaction to obtain chitosan - crosslinked nanospheres for standby; 6) Take 1 mL of the chitosan - crosslinked nanosphere solution in step 5) and add it to a microneedle mold, and place the mold in a centrifuge. Centrifuge at 4000 - 5000 r / min at 4 °C for 5 - 10 min to make the solution evenly fill the micropores of the mold; And place the molds uniformly in a freeze - dryer, and after drying, obtain soluble microneedles of urinary follicle - stimulating hormone combined with letrozole for injection.

2. The preparation method of the soluble microneedle of urofollitropin for injection combined with letrozole according to claim 1, wherein The method includes the following steps: 1) Dissolve 100 mg of poly (lactic - co - glycolic acid) in 1 mL of dichloromethane to obtain mixture 1 for standby; 2) Dissolve 1 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby; 3) Dissolve 2.5 mg of letrozole tablets in 2 mL of 75 IU urinary follicle - stimulating hormone for injection to obtain mixture 3 for standby; 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby; 5) Add the mixture 4 obtained in step 4) to the mixture 1 in step 1), and simultaneously add 500 mg of tripolyphosphate for cross - linking reaction to obtain chitosan - crosslinked nanospheres for standby; 6) Take 1 mL of the chitosan - crosslinked nanosphere solution in step 5) and add it to a microneedle mold, and place the mold in a centrifuge. Centrifuge at 5000 r / min at 4 °C for 5 min to make the solution evenly fill the micropores of the mold; And place the molds uniformly in a freeze - dryer, and after drying, obtain soluble microneedles of urinary follicle - stimulating hormone combined with letrozole for injection.

3. The preparation method of the soluble microneedle of urofollitropin for injection combined with letrozole according to claim 1, wherein The method includes the following steps: 1) Dissolve 150 mg of poly (lactic - co - glycolic acid) in 1.5 mL of dichloromethane to obtain mixture 1 for standby; 2) Dissolve 1.5 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby; 3) Dissolve 5 mg of letrozole tablets in 4 mL of 75 IU urinary follicle - stimulating hormone for injection to obtain mixture 3 for standby; 4) Add the mixture 3 obtained in step 3) to the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby; 5) Add the mixture 4 obtained in step 4) into the mixture 1 in step 1), and simultaneously add 800 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby; 6) Take 1 mL of the chitosan-crosslinked nanosphere solution in step 5) and add it into a microneedle mold, and place the mold in a centrifuge. Centrifuge at 5000 r / min at 4 °C for 5 min to make the solution uniformly fill the micropores of the mold; And place the molds uniformly in a freeze dryer, and obtain soluble microneedles of follicle-stimulating hormone for injection combined with letrozole after drying.

4. The preparation method of the soluble microneedle of urinary follicle-stimulating hormone for injection combined with letrozole according to claim 1, wherein, The method includes the following steps: 1) Dissolve 200 mg of poly (lactic-co-glycolic acid) copolymer in 2 mL of dichloromethane to obtain mixture 1 for standby; 2) Dissolve 2 g of chitosan in 0.1 M glacial acetic acid, and adjust the pH of the solution to 7.0 with NaOH to obtain mixture 2 for standby; 3) Dissolve 7.5 mg of letrozole tablets in 6 mL of 75 IU follicle-stimulating hormone for injection to obtain mixture 3 for standby; 4) Add the mixture 3 obtained in step 3) into the mixture 2 in step 2), and after mixing evenly, obtain mixture 4 for standby; 5) Add the mixture 4 obtained in step 4) into the mixture 1 in step 1), and simultaneously add 1000 mg of tripolyphosphate for cross-linking reaction to obtain chitosan-crosslinked nanospheres for standby; 6) Take 1 mL of the chitosan-crosslinked nanosphere solution in step 5) and add it into a microneedle mold, and place the mold in a centrifuge. Centrifuge at 5000 r / min at 4 °C for 5 min to make the solution uniformly fill the micropores of the mold; And place the molds uniformly in a freeze dryer, and obtain soluble microneedles of follicle-stimulating hormone for injection combined with letrozole after drying.

5. A soluble microneedle of urinary follicle-stimulating hormone for injection combined with letrozole, characterized in that, The soluble microneedles use PLGA-chitosan crosslinked microspheres as the microneedle body, and internally load follicle-stimulating hormone for injection and letrozole as active ingredients; wherein, the soluble microneedles are prepared by the method described in claim 1 above.

6. The soluble microneedle according to claim 5, wherein The length of the tip of the soluble microneedle is between 100 μm and 1 mm.

7. The soluble microneedle according to claim 6, wherein The tip of the microneedle may also be embedded with leuprolide acetate for injection, recombinant human follicle-stimulating hormone for injection, menotropins for injection, cetrorelix acetate for injection selected from.

8. Use of a soluble microneedle of urinary follicle-stimulating hormone combined with letrozole for injection in the preparation of a kit for treating infertility, characterized in that, The kit contains the soluble microneedles described in any one of claims 5-7.

9. The use according to claim 8, characterized in that, The administration method of the soluble microneedles is to directly attach the microneedles to the lower abdomen once a night for 5 consecutive days.

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