Pharmaceutical composition containing semen cuscutae and application thereof

Abstract

The invention discloses a pharmaceutical composition containing semen cuscutae, which is prepared from the following raw material medicines in parts by weight: 20 to 70 parts of semen cuscutae, 25 to 70 parts of fructus lycii, 10 to 35 parts of rhizoma dioscoreae, 0 to 20 parts of poria cocos and 1 to 20 parts of semen nelumbinis. The pharmaceutical composition can be used for preventing and / or treating ovarian reserve function decline, premature ovarian insufficiency and / or premature ovarian failure.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a pharmaceutical composition containing dodder seeds and a use thereof. Background Art

[0002] Ovarian reserve refers to the ability of follicles in the ovarian cortex to grow, develop, and form fertilizable and reproductive oocytes. Diminished ovarian reserve (DOR) is a process concept that covers multiple stages in the process of functional decline and includes both physiological and pathological aspects. With age, the ovarian function of older women gradually declines, and the hormone levels in the body smoothly transition to the menopausal state, which is physiological DOR. However, the ovarian reserve function of some women declines before the age of 40, presenting a pathological state that is inconsistent with age. The number and quality of mature oocytes produced by the ovaries decrease, and the clinical manifestations are irregular menstruation, amenorrhea, decreased fertility and even infertility. Modern Chinese medicine often classifies it as "late menstruation", "small menstruation", "amenorrhea" and "infertility". The incidence of DOR shows a trend of younger age, and the incidence rate is increasing year by year. At present, the incidence rate of DOR in the population is as high as 10-35%. DOR means that the reproductive function of women is aging prematurely. If not intervened in time, it is easy to develop into premature ovarian insufficiency (POI) or even premature ovarian failure (POF).

[0003] At present, hormone replacement therapy is used as a relatively standard Western medicine treatment model for the decline of ovarian reserve function. Progestogen and estrogen are mostly used clinically. Commonly used drugs are progesterone capsules, estradiol valerate (such as Progynova), and Fentaton. Studies have shown that Fentaton is a natural estrogen extracted from plants. It is a compound preparation of estradiol and dydrogesterone. It has the same structure as the active estrogen in the human body. It is a hormone drug for the treatment of ovarian dysfunction. It can be taken orally or vaginally. It is suitable for perimenopausal syndrome caused by natural or postoperative menopause. The research of Liu Meiyun and others showed that the therapeutic effect of Fentaton is better than artificial cycle treatment. Clinically, most patients regularly use estrogen-progesterone sequential therapy. After taking Fentaton (1 mg) orally, menstruation can come regularly, but the menstrual volume is small. Some studies have also shown that deoxyepiandrosterone (DHEA) can promote ovarian rejuvenation and increase the sensitivity of follicles to FSH stimulation. Yan Caixia et al. used DHEA and Progynova plus dydrogesterone tablets to treat DOR patients and found that DHEA can reduce serum FSH, increase AMH, increase AFC number, and improve ovarian reserve function, but there are also some corresponding side effects. Other studies have pointed out that bio-electric stimulation can cause resonance of cell biomembranes and surrounding macromolecules, increase energy, thereby promoting cell metabolism, accelerating blood flow, and increasing flow. This method can improve the blood supply to the ovaries and the surrounding areas of the ovaries, thereby achieving the purpose of improving ovarian function.

[0004] There is no related report in the prior art on the use of a pharmaceutical composition comprising dodder seeds as claimed in the present invention in preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure. Summary of the invention

[0005] Based on this, the present invention provides a pharmaceutical composition comprising dodder seeds. In parts by weight, the raw materials of the pharmaceutical composition include: 20-70 parts of dodder seeds, 25-70 parts of wolfberries, 10-35 parts of yam, 0-20 parts of poria cocos and 1-20 parts of lotus seeds.

[0006] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: about 48 parts of Cuscuta seeds, about 36 parts of Lycium barbarum, about 12 parts of Chinese yam and about 10.5 parts of lotus seeds.

[0007] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 43.2 to 52.8 parts of Cuscuta seeds, 32.4 to 39.6 parts of Lycium barbarum, 10.8 to 13.2 parts of Chinese yam and 9.45 to 11.55 parts of lotus seeds.

[0008] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 45.6-50.4 parts of Cuscuta seeds, 34.2-37.8 parts of Lycium barbarum, 11.4-12.6 parts of Chinese yam and 9.975-11.025 parts of lotus seeds.

[0009] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: about 54 parts of Cuscuta seeds, about 48 parts of Lycium barbarum, about 21 parts of Dioscorea opposita, about 7.5 parts of Poria cocos and about 4.5 parts of Lotus seeds.

[0010] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 48.6-59.4 parts of Cuscuta australis, 43.2-52.8 parts of Lycium barbarum, 18.9-23.1 parts of Dioscorea opposita, 6.75-8.25 parts of Poria cocos and 4.05-4.95 parts of Lotus seeds.

[0011] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 51.3 to 56.7 parts of Cuscuta seeds, 45.6 to 50.4 parts of Lycium barbarum, 19.95 to 22.05 parts of Dioscorea opposita, 7.125 to 7.875 parts of Poria cocos and 4.275 to 4.725 parts of Lotus seeds.

[0012] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: about 60 parts of Cuscuta seeds, about 60 parts of Lycium barbarum, about 30 parts of Dioscorea opposita, about 15 parts of Poria cocos and about 15 parts of Lotus seeds.

[0013] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 54-66 parts of Cuscuta seeds, 54-66 parts of Lycium barbarum, 27-33 parts of Dioscorea opposita, 13.5-16.5 parts of Poria cocos and 13.5-16.5 parts of Lotus seeds.

[0014] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 57-63 parts of Cuscuta seeds, 57-63 parts of Lycium barbarum, 28.5-31.5 parts of Dioscorea opposita, 14.25-15.75 parts of Poria cocos and 14.25-15.75 parts of Lotus seeds.

[0015] Furthermore, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is about 4: about 4: about 2: about 1: about 1.

[0016] Furthermore, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is (3.6-4.4): (3.6-4.4): (1.8-2.2): (0.9-1.1): (0.9-1.1).

[0017] Furthermore, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is (3.8-4.2): (3.8-4.2): (1.9-2.1): (0.95-1.05): (0.95-1.05).

[0018] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: about 27 parts of Cuscuta seeds, about 30 parts of Lycium barbarum, about 17 parts of Dioscorea opposita, about 12 parts of Poria cocos and about 14 parts of Lotus seeds.

[0019] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 24.3-29.7 parts of Cuscuta australis, 27-33 parts of Lycium barbarum, 15.3-18.7 parts of Dioscorea opposita, 10.8-13.2 parts of Poria cocos and 12.6-15.4 parts of Lotus seeds.

[0020] Furthermore, in parts by weight, the raw materials of the pharmaceutical composition include: 25.65-28.35 parts of Cuscuta australis, 28.5-31.5 parts of Lycium barbarum, 16.15-17.85 parts of Dioscorea opposita, 11.4-12.6 parts of Poria cocos and 13.3-14.7 parts of Lotus seeds.

[0021] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure.

[0022] Furthermore, the drug is selected from one or more of the following: progesterone capsules, estradiol valerate and fenmorphone.

[0023] Furthermore, the pharmaceutical composition is a traditional Chinese medicine composition.

[0024] According to another aspect of the present invention, there is provided a pharmaceutical preparation comprising the above pharmaceutical composition and pharmaceutically acceptable excipients.

[0025] Furthermore, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0026] Furthermore, the pharmaceutical preparation is in the form of pills, powders, tablets, drop pills, capsules, films, lozenges, granules, injections or oral solutions.

[0027] Furthermore, the pharmaceutical preparation is a pill.

[0028] According to another aspect of the present invention, there is provided a method for preparing the above-mentioned pharmaceutical composition, the method comprising the following steps:

[0029] (1) Weighing appropriate amounts of Cuscuta seeds, wolfberry fruits, Chinese yam, Poria cocos, and lotus seeds, mixing them evenly to obtain a mixture, adding water to boil, filtering, and obtaining a first filtrate and a residue;

[0030] (2) boiling the medicinal residue with water, filtering, and obtaining a second filtrate; and

[0031] (3) The first filtrate and the second filtrate are combined and concentrated to obtain the pharmaceutical composition.

[0032] Furthermore, in step (1), the volume / mass ratio of the water to the mixture is 8 to 12 times, for example, about 10 times.

[0033] Furthermore, the filtration is performed while hot.

[0034] Furthermore, the boiling time is 0.5 to 2 hours, for example, about 1 hour.

[0035] Furthermore, in step (2), the volume / mass ratio of the water to the medicinal residue is 8 to 12 times, for example, about 10 times.

[0036] Furthermore, the filtration is performed while hot.

[0037] Furthermore, the boiling time is 0.5 to 2 hours, for example, about 1 hour.

[0038] Furthermore, in step (3), the concentration is concentration under reduced pressure.

[0039] Furthermore, the concentration temperature is 30°C to 60°C, for example, about 40°C.

[0040] Furthermore, the concentration of the pharmaceutical composition is about 0.5-2 g of crude drug per ml, for example, about 1 g of crude drug.

[0041] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation in the preparation of a drug for preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure.

[0042] Furthermore, the decreased ovarian reserve function is pathological decreased ovarian reserve function.

[0043] Furthermore, the prevention and / or treatment of decreased ovarian reserve function is to promote the proliferation of human ovarian granulosa cells.

[0044] Furthermore, the human ovarian granulosa cells are KGN cells.

[0045] Furthermore, the decrease in ovarian reserve function is reflected in a decrease in ovarian estradiol levels.

[0046] Beneficial effects of the present invention:

[0047] The human ovarian granulosa cells (KGN cells) used in the present invention are currently the most commonly used cells for studying changes in ovarian function. Granulosa cells undergoing apoptosis or non-apoptotic programmed death are the main causes of follicular atresia. When follicular atresia intensifies, follicular development disorders will occur and the ovaries will age prematurely. The results of the CCK-8 experiment show that the composition can promote the proliferation of KGN cells, that is, it has an ovarian function protective effect and can prevent the occurrence of DOR. And through multiple regression analysis, it is concluded that when the composition has the strongest effect on promoting the proliferation of KGN cells, the prescription ratio of the composition is Cuscuta: Lycium barbarum: Chinese yam: Poria: Lotus seed = 27:30:17:12:14. In order to verify whether the screened prescription is the best prescription ratio of the composition, a comparative experiment was subsequently conducted on the composition 11 with the strongest effect on promoting the proliferation of KGN cells and the prescription screened by uniform design. The results of CCK8 and ELISA showed that the effect of composition 11 was more significant than that of the composition screened out by uniform design, so it was concluded that the optimal formula ratio of the composition was Cuscuta australis: Lycium barbarum: Chinese yam: Poria cocos: Lotus seed = 4:4:2:1:1. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0049] Figure 1 The results of the promoting effect of composition 1-11 on KGN cells are shown in FIG. ** P<0.01, *** P<0.001.

[0050] Figure 2 The results of the promoting effect of composition 1-11 on KGN cells at 10 μg / mL are shown in FIG. *** P<0.001.

[0051] Figure 3 The schematic diagram of the results of the promoting proliferation of KGN cells by the composition 11 of the present invention and the selected formula is shown in FIG. * P<0.05, ** P<0.01, *** P<0.001.

[0052] Figure 4 The effect of the selected composition 11 of the present invention on the E2 level of KGN cells is shown in the figure. Compared with the normal group: * P<0.05. DETAILED DESCRIPTION

[0053] The following will be combined with the drawings in 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 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 those skilled in the art without creative work are within the scope of protection of the present invention.

[0054] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0055] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0056] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0057] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0058] As described in the background technology section, the current hormone replacement therapy for decreased ovarian reserve function has certain toxic side effects and the prior art does not have any relevant reports on the prevention and / or treatment of decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure of a pharmaceutical composition comprising dodder seeds as claimed in the present invention. In order to solve the above problems, the present invention provides a pharmaceutical composition comprising dodder seeds, and the raw materials of the pharmaceutical composition include, by weight, 20 to 70 parts of dodder seeds, 25 to 70 parts of wolfberry fruits, 10 to 35 parts of yam, 0 to 20 parts of tuckahoe and 1 to 20 parts of lotus seeds.

[0059] In the present invention, when parts by weight, weight ratios, volume / mass ratios, mass, time, pressure, ratios, equivalents, concentrations, or other values ​​or parameters are expressed as ranges, preferred ranges, or a series of upper preferred values ​​and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is disclosed separately. For example, when the range "20-70" is disclosed, the described range should be interpreted as including the range "20-70", "20-65", "20-60", "20-55", "20-50", "20-45", "20-40", "20-35", "20-30", "20-25", "25-70", "25-65", "25-60", "25-55", "25-50", "25-45", "25-40", "25-35", "25-30", "30-70", etc. When a numerical range is described herein, unless otherwise specified, the range is intended to include its end values ​​and all integers and fractions within the range, and the technical effects of the present invention can be achieved within the above range.

[0060] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: about 48 parts of Cuscuta seeds, about 36 parts of Lycium barbarum, about 12 parts of Chinese yam and about 10.5 parts of lotus seeds.

[0061] In the present invention, "about" refers to a value within the range of ±10% of a specific value, preferably, a value within the range of ±5% of a specific value. For example, "about 48" includes ±10% of 48, or from 43.2 to 52.8, "about 48" includes ±5% of 48, or from 45.6 to 50.4; "about 36" includes ±10% of 36, or from 32.4 to 39.6, "about 36" includes ±5% of 36, or from 34.2 to 37.8; "about 12" includes ±10% of 12, or from 10.8 to 13.2, "about 12" includes ±5% of 12, or from 11.4 to 12.6; "about 10.5" includes ±10% of 10.5, or from 9.45 to 11.55, "about 10.5" includes ±5% of 10.5, or from 9.975 to 11.025.

[0062] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 43.2-52.8 parts of Cuscuta seeds, 32.4-39.6 parts of Lycium barbarum, 10.8-13.2 parts of Chinese yam and 9.45-11.55 parts of lotus seeds.

[0063] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 45.6-50.4 parts of Cuscuta seeds, 34.2-37.8 parts of Lycium barbarum, 11.4-12.6 parts of Chinese yam and 9.975-11.025 parts of lotus seeds.

[0064] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: about 54 parts of Cuscuta seeds, about 48 parts of Lycium barbarum, about 21 parts of Dioscorea opposita, about 7.5 parts of Poria cocos and about 4.5 parts of Lotus seeds.

[0065] In the present invention, "about" refers to a value within the range of ±10% of a specific value, preferably, a value within the range of ±5% of a specific value. For example, "about 54" includes ±10% of 54, or from 48.6 to 59.4, "about 54" includes ±5% of 54, or from 51.3 to 56.7; "about 48" includes ±10% of 48, or from 43.2 to 52.8, "about 48" includes ±5% of 48, or from 45.6 to 50.4; "about 21" includes ±10% of 21, or from 18.9 to 23.1, "about 21" includes ±10% of 21, or from 18.9 to 23.1. "About 7.5" includes ±10% of 7.5, or from 6.75 to 8.25, and "about 7.5" includes ±5% of 7.5, or from 7.125 to 7.875; "about 4.5" includes ±10% of 4.5, or from 4.05 to 4.95, and "about 4.5" includes ±5% of 4.5, or from 4.275 to 4.725.

[0066] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 48.6-59.4 parts of Cuscuta seeds, 43.2-52.8 parts of Lycium barbarum, 18.9-23.1 parts of Dioscorea opposita, 6.75-8.25 parts of Poria cocos and 4.05-4.95 parts of Lotus seeds.

[0067] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 51.3-56.7 parts of Cuscuta australis, 45.6-50.4 parts of Lycium barbarum, 19.95-22.05 parts of Dioscorea opposita, 7.125-7.875 parts of Poria cocos and 4.275-4.725 parts of Lotus seeds.

[0068] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: about 60 parts of Cuscuta seeds, about 60 parts of Lycium barbarum, about 30 parts of Dioscorea opposita, about 15 parts of Poria cocos and about 15 parts of Lotus seeds.

[0069] In the present invention, "about" refers to a value within the range of ±10% of a specific value, preferably, a value within the range of ±5% of a specific value. For example, "about 60" includes ±10% of 60, or from 54 to 66, "about 60" includes ±5% of 60, or from 57 to 63; "about 30" includes ±10% of 30, or from 27 to 33, "about 30" includes ±5% of 30, or from 28.5 to 31.5; "about 15" includes ±10% of 15, or from 13.5 to 16.5, "about 15" includes ±5% of 15, or from 14.25 to 15.75.

[0070] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 54-66 parts of Cuscuta seeds, 54-66 parts of Lycium barbarum, 27-33 parts of Dioscorea opposita, 13.5-16.5 parts of Poria cocos and 13.5-16.5 parts of Lotus seeds.

[0071] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 57-63 parts of Cuscuta seeds, 57-63 parts of Lycium barbarum, 28.5-31.5 parts of Dioscorea opposita, 14.25-15.75 parts of Poria cocos and 14.25-15.75 parts of Lotus seeds.

[0072] In a preferred embodiment, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is about 4: about 4: about 2: about 1: about 1.

[0073] In the present invention, "about" refers to a value within the range of ±10% of a specific value, preferably, a value within the range of ±5% of a specific value. For example, "about 4" includes ±10% of 4, or from 3.6 to 4.4, "about 4" includes ±5% of 4, or from 3.8 to 4.2; "about 2" includes ±10% of 2, or from 1.8 to 2.2, "about 2" includes ±5% of 2, or from 1.9 to 2.1; "about 1" includes ±10% of 1, or from 0.9 to 1.1, and "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0074] In a preferred embodiment, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is (3.6-4.4): (3.6-4.4): (1.8-2.2): (0.9-1.1): (0.9-1.1).

[0075] In a preferred embodiment, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds, wherein the weight ratio of dodder seed, wolfberry fruit, yam, poria cocos and lotus seeds is (3.8-4.2): (3.8-4.2): (1.9-2.1): (0.95-1.05): (0.95-1.05).

[0076] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: about 27 parts of Cuscuta seeds, about 30 parts of Lycium barbarum, about 17 parts of Dioscorea opposita, about 12 parts of Poria cocos and about 14 parts of Lotus seeds.

[0077] In the present invention, "about" refers to a value within a range of ±10% of a specific value, preferably, a value within a range of ±5% of a specific value. For example, “about 27” includes ±10% of 27, or from 24.3 to 29.7, and “about 27” includes ±5% of 27, or from 25.65 to 28.35; “about 30” includes ±10% of 30, or from 27 to 33, and “about 30” includes ±5% of 30, or from 28.5 to 31.5; “about 17” includes ±10% of 17, or from 15.3 to 18.7, and “about 17” includes ±5% of 17, or from 16.15 to 17.85; “about 12” includes ±10% of 12, or from 10.8 to 13.2, and “about 12” includes ±5% of 12, or from 11.4 to 12.6; “about 14” includes ±10% of 14, or from 12.6 to 15.4, and “about 14” includes ±5% of 14, or from 13.3 to 14.7.

[0078] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 24.3-29.7 parts of Cuscuta australis, 27-33 parts of Lycium barbarum, 15.3-18.7 parts of Dioscorea opposita, 10.8-13.2 parts of Poria cocos and 12.6-15.4 parts of Lotus seeds.

[0079] In a preferred embodiment, the raw materials of the pharmaceutical composition include, by weight: 25.65-28.35 parts of Cuscuta seeds, 28.5-31.5 parts of Lycium barbarum, 16.15-17.85 parts of Dioscorea opposita, 11.4-12.6 parts of Poria cocos and 13.3-14.7 parts of Lotus seeds.

[0080] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure.

[0081] In a preferred embodiment, the drug is selected from one or more of the following: progesterone capsules, estradiol valerate and fenmorphone.

[0082] In a preferred embodiment, the pharmaceutical composition is a traditional Chinese medicine composition.

[0083] According to another aspect of the present invention, there is provided a pharmaceutical preparation comprising the above pharmaceutical composition and pharmaceutically acceptable excipients.

[0084] In a preferred embodiment, the pharmaceutical preparation of the present invention contains at least one pharmaceutically acceptable excipient in an amount of 0.00001 to 50 wt.%, or 0.0001 to 10 wt.%, or 0.0001 to 5 wt.%, or 0.005 to 1 wt.%, or 0.1 to 20 wt.%, or 0.5 to 15 wt.%, or 1 to 5 wt.%, based on the weight of the pharmaceutical preparation.

[0085] In the present invention, the term "pharmaceutically acceptable" refers to a substance, such as a carrier or diluent, which does not abrogate the biological activity or properties of the compound and is relatively non-toxic, such as a substance that does not cause undesirable biological effects or interact in a deleterious manner with any of its components when administered to an individual.

[0086] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with subjects and active ingredients (i.e., capable of inducing the desired therapeutic effect without causing any undesirable local or systemic effects), which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995).

[0087] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0088] Those skilled in the art will know how to select specific chemical substances within the scope of the above-mentioned auxiliary material category.For example, the diluent can be selected from one or more of the following: icing sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate and calcium carbonate.The wetting agent can be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, cetyl alcohol, glycerol fatty acid ester (such as triacetin, glyceryl monostearate and the like), polyoxyethylene fatty acid ester, polyethylene glycol fatty acid ester, sodium lauryl sulfate, sorbitol fatty acid ester, sucrose fatty acid ester, polyoxyethylene ether, benzalkonium chloride, polyoxyethylene castor oil and docusate sodium.The adhesive can be selected from one or more of the following: polyvinyl pyrrolidone, hydroxypropyl cellulose, polyethylene glycol and methylcellulose. The disintegrant may be selected from one or more of the following: carboxymethylcellulose, carboxymethylcellulose calcium salt and sodium carboxymethylcellulose. The flavoring agent may be selected from one or more of the following: sorbitol, glucose, mannose, sucrose and lactose. The dispersant may be selected from one or more of the following: cross-linked sodium carboxymethylcellulose, sodium starch glycolate and pre-gelatinized corn starch. The plasticizer may be dibutyl sebacate and / or various citrates. The sustained-release agent may be selected from one or more of the following: sodium carboxymethylcellulose, low-substituted hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, hydroxyethylcellulose, gum arabic, gelatin and shellac. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium pyrosulfite, sodium sulfite and sodium thiosulfate. The lubricant may be selected from one or more of the following: calcium stearate, talc, magnesium stearate, stearic acid and colloidal silicon dioxide.

[0089] In a preferred embodiment, the pharmaceutical preparation is in the form of pills, powders, tablets, drop pills, capsules, films, lozenges, granules, injections or oral solutions.

[0090] In a preferred embodiment, the pharmaceutical preparation is a pill.

[0091] According to another aspect of the present invention, there is provided a method for preparing the above-mentioned pharmaceutical composition, the method comprising the following steps:

[0092] (1) Weighing appropriate amounts of Cuscuta seeds, wolfberry fruits, Chinese yam, Poria cocos, and lotus seeds, mixing them evenly to obtain a mixture, adding water to boil, filtering, and obtaining a first filtrate and a residue;

[0093] (2) boiling the medicinal residue with water, filtering, and obtaining a second filtrate; and

[0094] (3) combining the first filtrate and the second filtrate, and concentrating them to obtain the pharmaceutical composition.

[0095] In a preferred embodiment, in step (1), the volume / mass ratio of the water to the mixture is 8 to 12 times, for example, about 10 times.

[0096] In a preferred embodiment, the filtration is hot filtration.

[0097] In a preferred embodiment, the decocting time is 0.5 to 2 hours, for example, about 1 hour.

[0098] In a preferred embodiment, in step (2), the volume / mass ratio of the water to the medicinal residue is 8 to 12 times, for example, about 10 times.

[0099] In a preferred embodiment, the filtration is hot filtration.

[0100] In a preferred embodiment, the decocting time is 0.5 to 2 hours, such as about 1 hour.

[0101] In a preferred embodiment, in step (3), the concentration is concentration under reduced pressure.

[0102] In a preferred embodiment, the concentration temperature is 30°C to 60°C, such as about 40°C.

[0103] In a preferred embodiment, the concentration of the pharmaceutical composition is about 0.5-2 g of crude drug per ml, for example, about 1 g of crude drug.

[0104] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition or the above-mentioned pharmaceutical preparation in the preparation of a drug for preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure.

[0105] In the present invention, the term "treatment" also includes "prevention", unless there is a specific description to the contrary. The terms "treatment" and "therapeutically" should be understood accordingly.

[0106] In the present invention, the term "treatment" includes alleviating, inhibiting or improving the symptoms or conditions of a disease; inhibiting the occurrence of complications; improving or preventing potential metabolic syndrome; inhibiting the occurrence of a disease or symptom, such as controlling the development of a disease or condition; alleviating a disease or symptom; reducing a disease or symptom; alleviating complications caused by a disease or symptom, or preventing or treating signs caused by a disease or symptom. As used herein, a pharmaceutical composition or a pharmaceutical preparation, after administration, can improve a disease, symptom or condition, especially improve its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Whether fixed or temporary administration, continuous or intermittent administration, can be attributed to or related to the administration.

[0107] In a preferred embodiment, the decreased ovarian reserve is pathological decreased ovarian reserve.

[0108] In a preferred embodiment, the prevention and / or treatment of decreased ovarian reserve function is to promote the proliferation of human ovarian granulosa cells.

[0109] In a preferred embodiment, the human ovarian granulosa cells are KGN cells.

[0110] In a preferred embodiment, the decreased ovarian reserve function is a decrease in ovarian estradiol levels.

[0111] According to another aspect of the present invention, there is provided the above composition or the above pharmaceutical preparation for preventing and / or treating decreased ovarian reserve function in a subject.

[0112] According to another aspect of the present invention, there is provided a method for preventing and / or treating decreased ovarian reserve function in a subject, comprising administering an effective amount of the above composition or the above pharmaceutical preparation to the subject.

[0113] In the present invention, the term "subject" is a mammal. The mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse or a cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects representing the model of decreased ovarian reserve function. Preferably, the subject is a human.

[0114] The "effective amount" of the pharmaceutical composition or preparation used in the present invention can obtain the desired treatment and / or prevention effect. The amount effective for this purpose will depend on, for example, the pharmaceutical composition, the mode of administration, the stage and severity of the disease to be treated, the individual's body weight and overall health, and the judgment of the prescribing physician. The dosage can be given once a week, or once every two days or every day, or even several times a day. The dosage unit can be given in a short period of time (e.g., weeks to months) or longer time periods (months to years).

[0115] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0116] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0117] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0118] Example

[0119] Example 1 Uniform design screening of the formulation ratio of the composition

[0120] 1 Materials

[0121] 1.1 Drugs

[0122] Cuscuta australis (batch number: 220274), wolfberry (batch number: 22101704), yam (batch number: 21090701), poria (batch number: 23020401), and lotus seeds (batch number: 22031101) were purchased from Beijing Tong Ren Tang Suzhou Street Pharmaceutical Co., Ltd.

[0123] 1.2 Cells

[0124] Human ovarian granulosa cell line (KGN cells) was purchased from Wuhan Pronocell Life Science Technology Co., Ltd.

[0125] 1.3 Reagents

[0126] Fetal bovine serum, penicillin / streptomycin, DMEM / F12 medium, 0.25% trypsin, CCK-8

[0127] 2 Methods

[0128] 2.1 Drug combination and dosage ratio

[0129] This prescription is composed of 5 kinds of Chinese medicine. The dosage range of the 5 herbs in the 2020 edition of the Pharmacopoeia is referred to, and combined with clinical practice, the dosage range of each drug is determined. The 5 herbs are used as 5 factors and 11 levels are designed. The experiment adopts U 11 (11 5 ) uniform design table was used for arrangement, and the specific proportion relationship of the drugs in each dosage ratio group is shown in Table 1.

[0130] Table 1U 11 (11 5 )Uniform design table (unit: g)

[0131]

[0132] In order to eliminate the interference of dose-effect relationship on drug efficacy, the amount of raw drugs in each group was adjusted to the same dose (100 g) while keeping the drug ratio of each group unchanged. The adjusted drug amounts of each group are shown in Table 2.

[0133] Table 2 Composition of drugs in each group of the uniform composition test (unit: g)

[0134]

[0135]

[0136] 2.2 Drug preparation

[0137] The above 11 groups of medicinal materials were mixed evenly according to Table 2, added with 10 times the amount of water and boiled for 1 hour, filtered while hot, and the residues were added with 10 times the amount of water and boiled for 1 hour, filtered while hot, the two filtrates were combined, and concentrated under reduced pressure at 40°C to an extract containing 1 gram of crude drug per milliliter.

[0138] 2.3 Cell culture

[0139] KGN cells were cultured in a 37°C incubator containing 5% CO2 and saturated humidity in DMEM / F12 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin. The medium was changed every other day, and the cell growth status was observed under an inverted microscope. When the cells grew and merged into a dense monolayer (coverage rate reached 70%-80%), they were routinely digested with 0.25% trypsin, centrifuged at 1200 rpm for 3 minutes, and passaged once every 2-3 days at a ratio of 1:3 or 1:4.

[0140] 2.4CCK-8 detection of cell viability

[0141] In this experiment, different concentrations of composition 1-11 (0.1, 1, 10, 100 μg / mL) were used to intervene in KGN cells for 48 hours to observe the cell proliferation effect.

[0142] The specific steps are as follows:

[0143] (1) KGN cells in the logarithmic growth phase were taken and a single cell suspension was prepared using DMEM / F12 medium containing 10% fetal bovine serum. The cells were inoculated into a 96-well plate at a density of 5000 cells / well with a volume of 90 μL per well.

[0144] (2) The experiment was divided into a blank group, a normal control group, and different drug groups. The blank group was not inoculated with cells but only added with a culture medium of the same volume as the drug group. The normal control group was inoculated with cells and added with a culture medium of the same volume as the drug group. After the cells adhered to the wall, 10 μL of culture medium was added to the blank group and the normal control group, and 10 μL of the composition 1-11 corresponding to 10 times the concentration was added to each drug group. Three replicate wells were set up for each group. The final volume in each well was the same, all 100 μL;

[0145] (3) After 48 h of drug intervention, 10 μL of CCK-8 working solution was added to each well in the dark and incubated in a 5% CO2, 37°C cell culture incubator for 2 h; and

[0146] (4) Use an ELISA reader to measure the absorbance (OD value) of each well at a wavelength of 450 nm, and calculate the cell viability using the formula (OD value of the drug well - OD value of the blank well) / (OD value of the control well - OD value of the blank well).

[0147] 2.5 Statistical analysis

[0148] All data were analyzed using GraphPad Prism 9.5.0 statistical software. The data were analyzed by one-way analysis of variance (ANOVA), and P < 0.05 indicated that the difference was statistically significant. The uniform design experiment was analyzed by DPS quadratic polynomial stepwise regression.

[0149] 3 Results

[0150] 3.1 Each group of compositions has a promoting effect on KGN cell proliferation

[0151] Compared with the normal group, composition 9 (100 μg / mL), composition 10, and composition 11 (0.1, 1, 10, and 100 μg / mL) could significantly promote the proliferation of KGN cells (Table 3, Figure 1 ), taking the 10 μg / mL composition as an example, composition 11 has the best effect in promoting KGN cell proliferation ( Figure 2 ).

[0152] Table 3 Effects of compositions 1-11 on KGN cell viability

[0153]

[0154]

[0155] Note: Compared with the normal group: ** P<0.01, *** P<0.001.

[0156] 3.2 Analysis of uniform design results

[0157] After DPS quadratic polynomial stepwise regression analysis and combined with traditional Chinese medicine theory, the effect of 10 μg / mL composition on KGN cell viability (such as Figure 2 The regression equation is as shown in the figure: Y = -1147.895199 + 6.732529613 × X4 + 0.06974432564X2 × X2 - 2.0151764702X3 × X3 + 6.137745476X4 × X4 - 2.0307198676X5 × X5 + 1.3448226825X1 × X3 + 1.43 18536715X2×X3-0.7314223875X2×X5-5.193773299X3×X4+4.839879770X3×X5, Y is cell viability, X1 is Cuscuta, X2 is Lycium barbarum, X3 is Chinese yam, X4 is Poria, X5 is lotus seed, correlation coefficient R=0.8927, P<0.0001, the equation is significant. Theoretical maximum value Y max =3367.1987, at this time X1:X2:X3:X4:X5=27:30:17:12:14.

[0158] Embodiment 2

[0159] Effects of combination 11 (4:4:2:1:1) and the combination (27:30:17:12:14) screened by uniform design on KGN cell proliferation and estradiol (E2) level

[0160] 1 Materials

[0161] Human E2 enzyme-linked immunosorbent assay (ELISA) kit was purchased from Shanghai ELISA Biotechnology Co., Ltd. Other materials were the same as those in Example 1.

[0162] 2 Methods

[0163] 2.1 Drug preparation

[0164] Mix 27g of Cuscuta australis, 30g of Lycium barbarum, 17g of Chinese yam, 12g of Poria cocos and 14g of lotus seeds evenly, add 10 times the amount of water and boil for 1 hour, filter while hot, add 10 times the amount of water to the residue and boil for 1 hour, filter while hot, combine the two filtrates, and concentrate under reduced pressure at 40℃ to obtain an extract containing 1 gram of the crude drug per milliliter.

[0165] 2.2 Cell culture

[0166] Same as the first embodiment.

[0167] 2.3CCK-8 detection of cell viability

[0168] In this experiment, different concentrations of composition 11 and the formula screened out by uniform design method (0.1, 1, 10, 100 μg / mL) were used to intervene in KGN cells for 48 hours to observe the cell proliferation effect.

[0169] The specific steps are as follows:

[0170] (1) KGN cells in the logarithmic growth phase were taken and single-cell suspension was prepared using DMEM / F12 medium containing 10% fetal bovine serum. The cells were inoculated into a 96-well plate at a density of 5000 cells / well with a volume of 90 μL per well.

[0171] (2) The experiment was divided into a blank group, a normal control group and different drug groups. The blank group was not inoculated with cells but only added with the same volume of culture medium as the drug group. The normal control group was inoculated with cells and added with the same volume of culture medium as the drug group. After the cells adhered to the wall, 10 μL of culture medium was added to the blank group and the normal control group, and 10 μL of the corresponding 10-fold concentration of the composition 11 and the formula screened by the uniform design method was added to each drug group. Three replicate wells were set up for each group. The final volume in each well was the same, all 100 μL; and

[0172] (3) After 48 h of drug intervention, 10 μL of CCK-8 working solution was added to each well in the dark and incubated in a cell culture incubator with 5% CO2 and 37°C for 2 h. The absorbance (OD value) of each well was measured at a wavelength of 450 nm using an ELISA reader, and the cell viability was calculated using the formula (OD value of the drug well - OD value of the blank well) / (OD value of the control well - OD value of the blank well).

[0173] 2.4 ELISA to detect cellular E2 levels

[0174] In this experiment, 10 μg / mL composition 11 and the formula selected by uniform design method were used to intervene in KGN cells for 48 hours to detect the cellular E2 level.

[0175] The specific steps are as follows:

[0176] (1) KGN cells in the logarithmic growth phase were taken and single-cell suspension was prepared using phenol red-free DMEM / F12 medium containing 10% fetal bovine serum. The cells were seeded into a 96-well plate at a density of 5000 cells / well with a volume of 90 μL per well.

[0177] (2) After the cells adhered to the wall, 10 μL of culture medium was added to the normal control group, and 10 μL of the corresponding 10-fold concentration of composition 11 and the formula selected by the uniform design method was added to each drug group. Three replicate wells were set up for each group. The final volume in each well was the same, all 100 μL; and

[0178] (3) After 48 h of drug intervention, KGN cell supernatant was collected (centrifuged at 1000×g for 20 min) and tested according to the instructions of the human E2 ELISA kit.

[0179] 2.5 Statistical analysis

[0180] Same as the first embodiment.

[0181] 3 Results

[0182] 3.1 Proliferation-promoting effect of the combination 11 and the formula selected by uniform design on KGN cells

[0183] Compared with the normal group, composition 11 (0.1, 1, 10, 100 μg / mL) and the uniform design method screened formula (10, 100 μg / mL) can significantly promote the proliferation of KGN cells (Table 4, Figure 3 ), composition 11 has the best effect in promoting KGN cell proliferation.

[0184] Table 4 Effects of composition 11 and selected prescriptions on KGN cell viability

[0185]

[0186] 3.2 Effects of Composition 11 and the Formulas Screened by Uniform Design on Estradiol (E2) Levels in KGN Cells

[0187] like Figure 4 As shown, compared with the normal group, the E2 level of KGN cells treated with 10 μg / mL composition 11 was significantly increased (P<0.05), indicating that composition 11 can promote the secretion of E2 in KGN cells, while the composition 10 μg / mL screened by the uniform design method had no significant effect on the secretion of E2 in KGN cells.

[0188] 4 Conclusion

[0189] The human ovarian granulosa cells (KGN cells) used in the present invention are currently the most commonly used cells for studying changes in ovarian function. Granulosa cells undergoing apoptosis or non-apoptotic programmed death are the main causes of follicular atresia. When follicular atresia intensifies, follicular development disorders will occur and the ovaries will age prematurely. The results of the CCK-8 experiment show that the composition can promote the proliferation of KGN cells, that is, it has an ovarian function protective effect and can prevent the occurrence of DOR. And through multiple regression analysis, it is concluded that when the composition has the strongest effect on promoting the proliferation of KGN cells, the prescription ratio of the composition is Cuscuta: Lycium barbarum: Chinese yam: Poria: Lotus seed = 27:30:17:12:14. In order to verify whether the screened prescription is the best prescription ratio of the composition, a comparative experiment was subsequently conducted on the composition 11 with the strongest effect on promoting the proliferation of KGN cells and the prescription screened by uniform design. The results of CCK8 and ELISA showed that the effect of composition 11 was more significant than that of the composition screened out by uniform design, so it was concluded that the optimal formula ratio of the composition was Cuscuta australis: Lycium barbarum: Chinese yam: Poria cocos: Lotus seed = 4:4:2:1:1.

[0190] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A pharmaceutical composition comprising Cuscuta australis, characterized in that: The raw materials of the pharmaceutical composition include, by weight, 20 to 70 parts of dodder seeds, 25 to 70 parts of wolfberry fruits, 10 to 35 parts of yam, 0 to 20 parts of poria cocos and 1 to 20 parts of lotus seeds.

2. The pharmaceutical composition according to claim 1, characterized in that In parts by weight, the raw materials of the pharmaceutical composition include: about 48 parts of Cuscuta seeds, about 36 parts of Lycium barbarum, about 12 parts of Chinese yam and about 10.5 parts of lotus seeds; Preferably, the raw materials of the pharmaceutical composition include, by weight: 43.2-52.8 parts of Cuscuta seeds, 32.4-39.6 parts of Lycium barbarum, 10.8-13.2 parts of Chinese yam, and 9.45-11.55 parts of lotus seeds; Preferably, the raw materials of the pharmaceutical composition include, by weight: 45.6-50.4 parts of Cuscuta seeds, 34.2-37.8 parts of Lycium barbarum, 11.4-12.6 parts of Chinese yam, and 9.975-11.025 parts of lotus seeds; More preferably, the raw materials of the pharmaceutical composition include, by weight: about 54 parts of Cuscuta seeds, about 48 parts of Lycium barbarum, about 21 parts of Dioscorea opposita, about 7.5 parts of Poria cocos, and about 4.5 parts of Lotus seeds; More preferably, the raw materials of the pharmaceutical composition include, by weight: 48.6-59.4 parts of Cuscuta seeds, 43.2-52.8 parts of Lycium barbarum, 18.9-23.1 parts of Dioscorea opposita, 6.75-8.25 parts of Poria cocos, and 4.05-4.95 parts of Lotus seeds; More preferably, the raw materials of the pharmaceutical composition include, by weight: 51.3-56.7 parts of Cuscuta seeds, 45.6-50.4 parts of Lycium barbarum, 19.95-22.05 parts of Dioscorea opposita, 7.125-7.875 parts of Poria cocos, and 4.275-4.725 parts of Lotus seeds; Preferably, the raw materials of the pharmaceutical composition include, by weight: about 60 parts of Cuscuta seeds, about 60 parts of Lycium barbarum, about 30 parts of Dioscorea opposita, about 15 parts of Poria cocos and about 15 parts of Lotus seeds; Preferably, the raw materials of the pharmaceutical composition include, by weight: 54-66 parts of Cuscuta seeds, 54-66 parts of Lycium barbarum, 27-33 parts of Dioscorea opposita, 13.5-16.5 parts of Poria cocos and 13.5-16.5 parts of Lotus seeds; Preferably, the raw materials of the pharmaceutical composition include, by weight: 57-63 parts of Cuscuta seeds, 57-63 parts of Lycium barbarum, 28.5-31.5 parts of Dioscorea opposita, 14.25-15.75 parts of Poria cocos and 14.25-15.75 parts of Lotus seeds; Particularly preferably, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, Chinese yam, Poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, Chinese yam, Poria cocos and lotus seed is about 4: about 4: about 2: about 1: about 1; Particularly preferably, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, Chinese yam, poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, Chinese yam, poria cocos and lotus seed is (3.6-4.4): (3.6-4.4): (1.8-2.2): (0.9-1.1): (0.9-1.1); Particularly preferably, the raw materials of the pharmaceutical composition include dodder seed, wolfberry fruit, Chinese yam, poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, Chinese yam, poria cocos and lotus seed is (3.8-4.2): (3.8-4.2): (1.9-2.1): (0.95-1.05): (0.95-1.05); Particularly preferably, the raw materials of the pharmaceutical composition include, by weight: about 27 parts of Cuscuta seeds, about 30 parts of Lycium barbarum, about 17 parts of Dioscorea opposita, about 12 parts of Poria cocos and about 14 parts of Lotus seeds; Particularly preferably, the raw materials of the pharmaceutical composition include, by weight: 24.3-29.7 parts of Cuscuta seeds, 27-33 parts of Lycium barbarum, 15.3-18.7 parts of Dioscorea opposita, 10.8-13.2 parts of Poria cocos, and 12.6-15.4 parts of Lotus seeds; Particularly preferably, the raw materials of the pharmaceutical composition include, by weight: 25.65-28.35 parts of Cuscuta seeds, 28.5-31.5 parts of Lycium barbarum, 16.15-17.85 parts of Dioscorea opposita, 11.4-12.6 parts of Poria cocos and 13.3-14.7 parts of Lotus seeds.

3. The pharmaceutical composition according to claim 1 or 2, characterized in that The pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and / or treating decreased ovarian reserve, premature ovarian insufficiency and / or premature ovarian failure; Preferably, the drug is selected from one or more of the following: progesterone capsules, estradiol valerate and fenmorphone; More preferably, the pharmaceutical composition is a traditional Chinese medicine composition.

4. A pharmaceutical preparation comprising the pharmaceutical composition according to any one of claims 1 to 3 and a pharmaceutically acceptable excipient.

5. The pharmaceutical preparation according to claim 4, characterized in that The auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant; Preferably, the pharmaceutical preparation is a pill, powder, tablet, drop pill, capsule, film, lozenge, granule, injection or oral solution; More preferably, the pharmaceutical preparation is a pill.

6. A method for preparing the pharmaceutical composition according to claim 1 or 2, characterized in that: The method comprises the following steps: (1) Weighing appropriate amounts of Cuscuta seeds, wolfberry fruits, Chinese yam, Poria cocos, and lotus seeds, mixing them evenly to obtain a mixture, adding water to boil, filtering, and obtaining a first filtrate and a residue; (2) boiling the medicinal residue with water, filtering and obtaining a second filtrate; as well as (3) combining the first filtrate and the second filtrate, and concentrating them to obtain the pharmaceutical composition.

7. The method according to claim 6, characterized in that In step (1), the volume / mass ratio of the water to the mixture is 8 to 12 times, for example, about 10 times; Preferably, the filtration is hot filtration; More preferably, the decocting time is 0.5 to 2 hours, such as about 1 hour.

8. The method according to claim 6, characterized in that In step (2), the volume / mass ratio of the water to the medicinal residue is 8 to 12 times, for example, about 10 times; Preferably, the filtration is hot filtration; More preferably, the decocting time is 0.5 to 2 hours, such as about 1 hour.

9. The method according to claim 6, characterized in that In step (3), the concentration is concentration under reduced pressure; Preferably, the concentration temperature is 30°C to 60°C, for example about 40°C; More preferably, the concentration of the pharmaceutical composition is about 0.5 to 2 g of the crude drug per ml, for example, about 1 g of the crude drug.

10. Use of the pharmaceutical composition according to any one of claims 1 to 3 or the pharmaceutical preparation according to claim 4 or 5 in the preparation of a medicament for preventing and / or treating decreased ovarian reserve function, premature ovarian insufficiency and / or premature ovarian failure; Preferably, the decreased ovarian reserve function is pathological decreased ovarian reserve function; More preferably, the prevention and / or treatment of decreased ovarian reserve function is to promote the proliferation of human ovarian granulosa cells; Also preferably, the human ovarian granulosa cells are KGN cells; Particularly preferably, the decreased ovarian reserve function is a decrease in ovarian estradiol levels.

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