A pharmaceutical composition comprising cuscuta and uses thereof

Abstract

The application discloses a kind of medicine compositions comprising semya, and the raw materials of the medicine compositions include, by weight parts: semya 20-70 parts, medlar 25-70 parts, Chinese yam 10-35 parts, tuckahoe 0-20 parts and lotus seed 1-20 parts.The medicine compositions of the application can be used for preventing and / or treating decreased ovarian reserve, premature ovarian insufficiency and / or premature ovarian failure.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more specifically to a pharmaceutical composition containing dodder seed and its uses. Background Technology

[0002] Ovarian reserve refers to the ability of follicles in the ovarian cortex to grow, develop, and form fertile, reproductive oocytes. Diminished ovarian reserve (DOR) is a process-related concept encompassing multiple stages of functional decline, including both physiological and pathological aspects. With age, ovarian function gradually declines in older women, and hormone levels smoothly transition towards menopause—this is physiological DOR. However, some women experience a decline in ovarian reserve before age 40, exhibiting a pathological state inconsistent with their age. This results in a reduced number and quality of mature oocytes produced by the ovaries, clinically manifesting as menstrual irregularities, amenorrhea, decreased fertility, and even infertility. Modern Traditional Chinese Medicine often categorizes this as "delayed menstruation," "scanty menstruation," "amenorrhea," or "infertility." The onset of DOR is showing a trend towards younger ages, and the incidence rate is increasing annually, currently ranging from 10% to 35% in the population. DOR signifies premature aging of female reproductive function; without timely intervention, it can easily develop into premature ovarian insufficiency (POI) or even premature ovarian failure (POF).

[0003] Currently, hormone replacement therapy is the standard Western medicine treatment for declining ovarian reserve. Clinically, progesterone and estrogen are commonly used. Commonly used drugs include progesterone capsules, estradiol valerate (such as Progynova), and Femoston. Studies have shown that Femoston is a natural estrogen extracted from plants, a compound preparation of estradiol and dydrogesterone, structurally identical to active estrogen in the human body. It is a hormonal drug for treating ovarian dysfunction, and can be administered orally or vaginally, suitable for perimenopausal syndrome caused by natural or postoperative menopause. Research by Liu Meiyun et al. shows that the therapeutic effect of Femoston is superior to artificial cycle therapy. Clinically, most patients who regularly use sequential estrogen-progesterone therapy experience regular menstruation after taking Femoston (1mg), although the menstrual flow is often light. Studies have also shown that dehydroepiandrosterone (DHEA) can promote ovarian rejuvenation and increase the sensitivity of follicles to FSH stimulation. Yan Caixia et al. treated DOR patients with DHEA, estradiol valerate, and dydrogesterone tablets and found that DHEA could lower serum FSH, increase AMH, and increase AFC count, thus improving ovarian reserve function, but also had some corresponding side effects. Other studies have indicated that biomimetic electrical stimulation can cause resonance in cell membranes and surrounding macromolecules, increasing energy and thus promoting cell metabolism, accelerating blood flow, and increasing blood volume. This method can improve the blood supply to the ovaries and surrounding tissues, thereby improving ovarian function.

[0004] There are no existing reports on the use of the pharmaceutical composition containing dodder seed claimed in this invention for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency and / or premature ovarian failure. Summary of the Invention

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

[0006] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: approximately 48 parts of Cuscuta chinensis, approximately 36 parts of Lycium barbarum, approximately 12 parts of Dioscorea opposita, and approximately 10.5 parts of Nelumbo nucifera.

[0007] Furthermore, by weight, the active pharmaceutical ingredients of the pharmaceutical composition include: 43.2–52.8 parts of Cuscuta chinensis, 32.4–39.6 parts of Lycium barbarum, 10.8–13.2 parts of Dioscorea opposita, and 9.45–11.55 parts of Nelumbo nucifera.

[0008] Furthermore, by weight, the active pharmaceutical ingredients of the pharmaceutical composition include: 45.6–50.4 parts of Cuscuta chinensis, 34.2–37.8 parts of Lycium barbarum, 11.4–12.6 parts of Dioscorea opposita, and 9.975–11.025 parts of Nelumbo nucifera.

[0009] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: approximately 54 parts of Cuscuta chinensis, approximately 48 parts of Lycium barbarum, approximately 21 parts of Dioscorea opposita, approximately 7.5 parts of Poria cocos, and approximately 4.5 parts of Nelumbo nucifera.

[0010] Furthermore, by weight, the active pharmaceutical ingredients of the pharmaceutical composition include: 48.6–59.4 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0011] Furthermore, by weight, the active pharmaceutical ingredients of the pharmaceutical composition include: 51.3–56.7 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0012] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: approximately 60 parts of Cuscuta chinensis, approximately 60 parts of Lycium barbarum, approximately 30 parts of Dioscorea opposita, approximately 15 parts of Poria cocos, and approximately 15 parts of Nelumbo nucifera.

[0013] Furthermore, by weight, the raw materials of the pharmaceutical composition include: 54-66 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0014] Furthermore, by weight, the raw materials of the pharmaceutical composition include: 57-63 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

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

[0016] Furthermore, the active ingredients of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, 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).

[0017] Furthermore, the active ingredients of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, 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).

[0018] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: approximately 27 parts of Cuscuta chinensis, approximately 30 parts of Lycium barbarum, approximately 17 parts of Dioscorea opposita, approximately 12 parts of Poria cocos, and approximately 14 parts of Nelumbo nucifera.

[0019] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: 24.3–29.7 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0020] Furthermore, by weight, the active ingredients of the pharmaceutical composition include: 25.65–28.35 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0021] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for the prevention and / or treatment of decreased ovarian reserve, 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 femoston.

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

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

[0025] Furthermore, the excipient is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, encapsulating agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers, and antioxidants.

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

[0027] Furthermore, the drug formulation is in the form of pills.

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

[0029] (1) Weigh out appropriate amounts of dodder seed, wolfberry, yam, poria cocos and lotus seed, mix them evenly to obtain a mixture, add water to decoct, filter, and obtain the first filtrate and dregs;

[0030] (2) Boil the dregs in water, filter, and obtain a second filtrate; and

[0031] (3) Combine the first filtrate and the second filtrate, concentrate them, and obtain the drug 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 the filter is still hot.

[0034] Furthermore, the boiling time with water 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 dregs is 8 to 12 times, for example, about 10 times.

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

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

[0038] Furthermore, in step (3), the concentration is a vacuum concentration.

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

[0040] Furthermore, the concentration of the pharmaceutical composition is approximately 0.5 to 2 g of crude drug per milliliter, for example, approximately 1 g of crude drug.

[0041] According to another aspect of the present invention, there is provided the use of the above-described pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency and / or premature ovarian failure.

[0042] Furthermore, this decline in ovarian reserve is termed pathological ovarian reserve decline.

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

[0044] Furthermore, the ovarian granulosa cells in this individual were KGN cells.

[0045] Furthermore, this decline in ovarian reserve is manifested by a decrease in ovarian estradiol levels.

[0046] The beneficial effects of this invention are:

[0047] This invention utilizes human ovarian granulosa cells (KGN cells), currently the most commonly used cell type for studying changes in ovarian function. Apoptosis or non-apoptotic programmed cell death in granulosa cells is a major cause of follicular atresia; severe follicular atresia leads to follicular developmental disorders and premature ovarian aging. CCK-8 assay results show that the composition can promote KGN cell proliferation, thus exhibiting ovarian function protection and preventing ovarian dysfunction (DOR). Furthermore, multivariate regression analysis revealed that the optimal ratio of the composition for promoting KGN cell proliferation was 27:30:17:12:14 (Cuscuta chinensis: Lycium barbarum: Dioscorea opposita: Poria cocos: Nelumbo nucifera). To verify whether the selected formulation represents the optimal ratio, a comparative experiment was conducted between the composition 11, which showed the strongest effect in promoting KGN cell proliferation, and the formulation selected using a uniform design. The results of CCK8 and ELISA showed that composition 11 was more effective than the composition selected by uniform design. Therefore, it was concluded that the optimal formulation ratio of the composition was Cuscuta chinensis: Lycium barbarum: Dioscorea opposita: Poria cocos: Nelumbo nucifera = 4:4:2:1:1. Attached Figure Description

[0048] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without exceeding the scope of protection claimed by the present invention.

[0049] Figure 1 This is a schematic diagram showing the proliferative effect of composition 1-11 on KGN cells. The comparison with the normal group is as follows: ** P<0.01, *** P<0.001.

[0050] Figure 2 This is a schematic diagram illustrating the proliferative effect of composition 1-11 (10 μg / mL) on KGN cells. The comparison with the normal group is shown below. *** P<0.001.

[0051] Figure 3 This is a schematic diagram showing the effect of composition 11 of the present invention on the proliferative activity of KGN cells, specifically the screened formulation. Compared with the normal group: * P<0.05, ** P<0.01, *** P<0.001.

[0052] Figure 4 This diagram illustrates the effect of composition 11 of the present invention and the screened formulations on E2 levels in KGN cells. Compared to the normal group: * P<0.05. Detailed Implementation

[0053] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

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

[0055] This invention is intended to cover all options, variations, and equivalents that may be included in the field of prior art as defined in the claims. Those skilled in the art will recognize many similar or equivalent methods and substances described herein that can be applied in the practice of this invention. This invention is by no means limited to the description of methods and substances.

[0056] The singular forms “a,” “an,” and “the” used in the specification and appended claims include plural indicators unless the context clearly specifies otherwise.

[0057] In this invention, the term "comprising" and "including" are synonymous. The terms "comprising," "including," "having," "containing," or any other variations thereof as used herein are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that includes the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such a composition, step, method, article, or apparatus.

[0058] As described in the background section, current hormone replacement therapy for decreased ovarian reserve has certain toxic side effects, and there are no existing reports on the use of the pharmaceutical composition containing dodder seed claimed in this invention for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency, and / or premature ovarian failure. To address the above problems, this invention provides a pharmaceutical composition containing dodder seed, wherein, by weight, the raw materials of the pharmaceutical composition include: 20-70 parts dodder seed, 25-70 parts wolfberry fruit, 10-35 parts yam, 0-20 parts poria cocos, and 1-20 parts lotus seed.

[0059] In this invention, when parts by weight, weight ratio, volume / mass ratio, mass, time, pressure, proportion, equivalent, concentration, or other values ​​or parameters are expressed as ranges, preferred ranges, or a series of upper and lower preferred values, this should be understood as specifically disclosing all ranges formed by any pairing of any upper or preferred value with any lower or preferred value, regardless of whether the range is disclosed individually. For example, when the range "20-70" is disclosed, the described range should be interpreted as including ranges "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 range of values ​​is described herein, unless otherwise stated, the range is intended to include its endpoints and all integers and fractions within that range, and the technical effects of the present invention can be achieved within the aforementioned range.

[0060] In a preferred embodiment, the active pharmaceutical ingredients of the pharmaceutical composition, by weight, include: about 48 parts of Cuscuta chinensis, about 36 parts of Lycium barbarum, about 12 parts of Dioscorea opposita, and about 10.5 parts of Nelumbo nucifera.

[0061] In this invention, "about" refers to a value within ±10% of a specific value, preferably within ±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 active pharmaceutical ingredients of the pharmaceutical composition, by weight, include: 43.2-52.8 parts of Cuscuta chinensis, 32.4-39.6 parts of Lycium barbarum, 10.8-13.2 parts of Dioscorea opposita, and 9.45-11.55 parts of Nelumbo nucifera.

[0063] In a preferred embodiment, the active pharmaceutical ingredients of the pharmaceutical composition, by weight, include: 45.6-50.4 parts of Cuscuta chinensis, 34.2-37.8 parts of Lycium barbarum, 11.4-12.6 parts of Dioscorea opposita, and 9.975-11.025 parts of Nelumbo nucifera.

[0064] In a preferred embodiment, the active pharmaceutical ingredients of the pharmaceutical composition, by weight, include: about 54 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0065] In this invention, "about" refers to a value within ±10% of a specific value, preferably within ±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. "Approximately 7.5" includes 21 ± 5%, or from 19.95 to 22.05; "approximately 7.5" includes 7.5 ± 10%, or from 6.75 to 8.25; "approximately 7.5" includes 7.5 ± 5%, or from 7.125 to 7.875; "approximately 4.5" includes 4.5 ± 10%, or from 4.05 to 4.95; "approximately 4.5" includes 4.5 ± 5%, or from 4.275 to 4.725.

[0066] In a preferred embodiment, the active ingredients of the pharmaceutical composition, by weight, include: 48.6-59.4 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0067] In a preferred embodiment, the active pharmaceutical ingredients of the pharmaceutical composition, by weight, include: 51.3-56.7 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0068] In a preferred embodiment, the active ingredients of the pharmaceutical composition, by weight, include: about 60 parts of Cuscuta chinensis, about 60 parts of Lycium barbarum, about 30 parts of Dioscorea opposita, about 15 parts of Poria cocos, and about 15 parts of Nelumbo nucifera.

[0069] In this 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 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 active ingredients of the pharmaceutical composition, by weight, include: 54-66 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0071] In a preferred embodiment, the raw materials of the pharmaceutical composition, by weight, include: 57-63 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

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

[0073] In this invention, "about" refers to a value within ±10% of a specific value, preferably within ±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; "about 1" includes ±5% of 1, or from 0.95 to 1.05.

[0074] In a preferred embodiment, the active pharmaceutical ingredients of the pharmaceutical composition include Cuscuta chinensis, Lycium barbarum, Dioscorea opposita, Poria cocos, and Nelumbo nucifera, wherein the weight ratio of Cuscuta chinensis, Lycium barbarum, Dioscorea opposita, Poria cocos, and Nelumbo nucifera 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 active pharmaceutical ingredients of the pharmaceutical composition include dodder seed, wolfberry fruit, yam, poria cocos and lotus seed, wherein the weight ratio of dodder seed, wolfberry fruit, 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).

[0076] In a preferred embodiment, the active ingredients of the pharmaceutical composition, by weight, include: about 27 parts of Cuscuta chinensis, about 30 parts of Lycium barbarum, about 17 parts of Dioscorea opposita, about 12 parts of Poria cocos, and about 14 parts of Nelumbo nucifera.

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

[0078] In a preferred embodiment, the active ingredients of the pharmaceutical composition, by weight, include: 24.3-29.7 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0079] In a preferred embodiment, the active ingredients of the pharmaceutical composition, by weight, include: 25.65-28.35 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

[0080] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for the prevention and / or treatment of decreased ovarian reserve, 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 femoston.

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

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

[0084] In a preferred embodiment, the pharmaceutical formulation 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.% relative to the weight of the pharmaceutical formulation.

[0085] In this invention, the term "pharmaceutically acceptable" means a substance, such as a carrier or diluent, that does not diminish the biological activity or properties of a compound and is relatively non-toxic, such that, if administered to an individual, it will not cause unwanted biological effects or interact with any of its constituent components in a harmful manner.

[0086] In this invention, the term "pharmaceuticalally acceptable excipient" refers to a carrier and / or excipient that is pharmacologically and / or physiologically compatible with the subject and the active ingredient (i.e., capable of eliciting the desired therapeutic effect without causing any undesirable local or systemic effects), which is 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 excipient is selected from one or more of the following: diluents, wetting agents, binders, disintegrants, encapsulating agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, light-blocking agents, and antioxidants.

[0088] Those skilled in the art will know how to select specific chemical substances within the aforementioned excipient categories. For example, the diluent may be selected from one or more of the following: powdered sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate, and calcium carbonate. The wetting agent may be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ethers, polysorbate esters such as polysorbate 80, cetyl alcohol, glyceryl fatty acid esters (such as triacetin, glyceryl monostearate, and the like), polyoxyethylene fatty acid esters, polyethylene glycol fatty acid esters, sodium lauryl sulfate, sorbitol fatty acid esters, sucrose fatty acid esters, polyoxyethylene ethers, benzalkonium chloride, polyoxyethylene castor oil, and sodium docusate. The binder may be selected from one or more of the following: polyvinylpyrrolidone, hydroxypropyl cellulose, polyethylene glycol, and methylcellulose. The disintegrant may be selected from one or more of the following: carboxymethyl cellulose, calcium carboxymethyl cellulose, and sodium carboxymethyl cellulose. 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: croscarmellose sodium, sodium starch glycolate, and pregelatinized corn starch. The plasticizer may be dibutyl sebacate and / or various citrate esters. The sustained-release agent may be selected from one or more of the following: sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methyl cellulose, microcrystalline cellulose, hydroxyethyl cellulose, gum arabic, gelatin, and shellac. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium metabisulfite, 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 silica.

[0089] In a preferred embodiment, the pharmaceutical preparation is a pill, powder, tablet, drop pill, capsule, film, lozenge, granule, injection, or oral liquid.

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

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

[0092] (1) Weigh out appropriate amounts of dodder seed, wolfberry, yam, poria cocos and lotus seed, mix them evenly to obtain a mixture, add water to decoct, filter, and obtain the first filtrate and dregs;

[0093] (2) Boil the dregs in water, filter, and obtain a second filtrate; and

[0094] (3) Combine the first filtrate and the second filtrate, concentrate them, and obtain the drug 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 performed while the material is still hot.

[0097] In a preferred embodiment, the boiling 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 dregs is 8 to 12 times, for example, about 10 times.

[0099] In a preferred embodiment, the filtration is performed while the material is still hot.

[0100] In a preferred embodiment, the boiling time is 0.5 to 2 hours, for example, about 1 hour.

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

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

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

[0104] According to another aspect of the present invention, there is provided the use of the above-described pharmaceutical composition or pharmaceutical preparation in the preparation of a medicament for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency and / or premature ovarian failure.

[0105] In this invention, the term "treatment" also includes "prevention," unless specifically stated otherwise. The terms "therapeutic" and "therapeutically" should be understood accordingly.

[0106] In this invention, the term "treatment" includes alleviating, suppressing, or improving symptoms or conditions of a disease; suppressing the development of complications; improving or preventing underlying metabolic syndromes; suppressing the development of a disease or symptom, such as controlling the progression of a disease or condition; reducing a disease or symptom; alleviating a disease or symptom; reducing complications arising from a disease or symptom; or preventing or treating signs arising from a disease or symptom. As used herein, a pharmaceutical composition or pharmaceutical preparation, upon administration, may improve a disease, symptom, or condition, particularly by improving its severity, delaying onset, slowing disease progression, or reducing the duration of the condition. Whether the administration is fixed or intermittent, continuous or intermittent, it may be attributable to or related to the administration.

[0107] In a preferred embodiment, the decline in ovarian reserve is a pathological decline in 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 decline in ovarian reserve is defined as a decrease in ovarian estradiol levels.

[0111] According to another aspect of the present invention, the above-described composition or pharmaceutical preparation is provided for the prevention and / or treatment of decreased ovarian reserve in a subject.

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

[0113] In this invention, the term "subject" refers to a mammal. A mammal can 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 advantageously be used as subjects representing a model of declining ovarian reserve. Preferably, the subject is a human.

[0114] The “effective amount” of the pharmaceutical composition or formulation used in this invention can achieve the desired therapeutic and / or preventative effects. The effective amount for this purpose will depend on factors such as the pharmaceutical composition, the route of administration, the stage and severity of the disease being treated, the individual’s weight and overall health, and the judgment of the prescribing physician. The dose can be administered once a week, every two days, or daily, or even several times a day. Dosage units can be administered over a short period (e.g., weeks to months) or a longer period (months to years).

[0115] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Experimental methods in the following embodiments, unless otherwise specified, are generally performed under conventional conditions or conditions recommended by the manufacturer.

[0116] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as are familiar to those skilled in the art. Furthermore, any methods and materials similar to or equivalent to those described herein may be applied to the methods of this invention. The preferred embodiments and materials described herein are for illustrative purposes only.

[0117] The features mentioned above in this invention, or the features mentioned in the embodiments, can be combined arbitrarily. All features disclosed in this patent specification can be used in any compositional form, and each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equivalent, or similar purpose. Therefore, unless otherwise specified, the disclosed features are merely general examples of equivalent or similar features.

[0118] Example

[0119] Example 1: Uniform Design of the Formulation Ratio of the Screening Composition

[0120] 1. Materials

[0121] 1.1 Drugs

[0122] Cuscuta chinensis (batch number: 220274), Lycium barbarum (batch number: 22101704), Dioscorea opposita (batch number: 21090701), Poria cocos (batch number: 23020401), and Nelumbo nucifera (batch number: 22031101) were all purchased from Beijing Tongrentang Suzhou Street Pharmaceutical Co., Ltd.

[0123] 1.2 Cells

[0124] Human ovarian granulosa cell line (KGN cells) was purchased from Wuhan Pronosei 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 formula consists of five Chinese medicinal herbs. Referring to the prescribed dosage ranges for these five herbs in the 2020 edition of the Chinese Pharmacopoeia, and combining this with clinical practice, the dosage ranges for each herb were determined. The five herbs were treated as five factors, and an 11-level experimental design was implemented. The experiment used a U0.05... 11 (11 5 The drugs were arranged in a uniform design table, and the specific proportions of each dosage ratio group are shown in Table 1.

[0130] Table 1U 11 (11 5 Uniform Design Table (Unit: g)

[0131]

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

[0133] Table 2. Drug composition of each group in the homogenization experiment (unit: g)

[0134]

[0135]

[0136] 2.2 Drug Preparation

[0137] The above 11 groups of medicinal materials are mixed evenly according to Table 2, decocted with 10 times the amount of water for 1 hour, filtered while hot, and the residue is decocted with 10 times the amount of water for 1 hour, filtered while hot, and the two filtrates are combined and concentrated under reduced pressure at 40℃ to an extract containing 1 gram of raw herb per milliliter.

[0138] 2.3 Cell Culture

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

[0140] 2.4 CCK-8 assay for cell viability

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

[0142] The specific steps are as follows:

[0143] (1) Take KGN cells in the logarithmic growth phase, prepare a single-cell suspension in DMEM / F12 medium containing 10% fetal bovine serum, and seed them into 96-well plates 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 was given the same volume of culture medium as the drug group. The normal control group was inoculated with cells and given the same volume of culture medium as the drug group. After the cells adhered, 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 combination 1-11 was added to each drug group. Each group was divided into 3 replicates. The final volume in each well was the same, which was 100 μL.

[0145] (3) After 48 hours of drug intervention, 10 μL of CCK-8 working solution was added to each well in the dark, and the cells were incubated for 2 hours in a 5% CO2, 37°C cell culture incubator; 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. The calculation formula is (OD value of drug well - OD value of blank well) / (OD value of control well - OD value of blank well).

[0147] 2.5 Statistical Analysis

[0148] All data were statistically analyzed using GraphPad Prism 9.5.0 software. Experimental results are presented as mean ± standard deviation. The results indicate that the analysis was performed using one-way ANOVA, with P < 0.05 indicating statistical significance. The uniform design experiment was analyzed using DPS quadratic polynomial stepwise regression analysis.

[0149] 3 Results

[0150] 3.1 Each composition group promoted the proliferation of KGN cells.

[0151] Compared with the normal group, composition 9 (100 μg / mL), compositions 10, and 11 (0.1, 1, 10, and 100 μg / mL) significantly promoted KGN cell proliferation (Table 3). Figure 1 Taking a 10 μg / mL composition as an example, composition 11 showed 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 stepwise regression analysis using DPS quadratic polynomials, combined with traditional Chinese medicine theory, the effect of a 10 μg / mL composition on KGN cell viability was ultimately determined (e.g., Figure 2 (As shown) The regression equation is: Y = -1147.895199 + 6.732529613 × X4 + 0.06974432564X2 × X2 - 2.0151764702X3 × X3 + 6.137745476X4 × X4 - 2.0307198676X5 × X5 + 1.3448226825X1 × X3 + 1.43 The equation is: 18536715X2×X3-0.7314223875X2×X5-5.193773299X3×X4+4.839879770X3×X5, where Y represents cell viability, X1 is Cuscuta chinensis, X2 is Lycium barbarum, X3 is Dioscorea opposita, X4 is Poria cocos, and X5 is Nelumbo nucifera. The correlation coefficient R = 0.8927, P < 0.0001, indicating the equation is statistically significant. The theoretical maximum value Y... max =3367.1987, at this time X1:X2:X3:X4:X5 =27:30:17:12:14.

[0158] Example 2

[0159] Effects of composition 11 (4:4:2:1:1) and the formulation selected by uniform design method (27:30:17:12:14) on the proliferation capacity and estradiol (E2) level of KGN cells.

[0160] 1. Materials

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

[0162] 2 Methods

[0163] 2.1 Drug Preparation

[0164] Mix 27g of Cuscuta chinensis, 30g of Lycium barbarum, 17g of Dioscorea opposita, 12g of Poria cocos, and 14g of Nelumbo nucifera seeds evenly, add 10 times the amount of water and decoct for 1 hour. Filter while hot. Add 10 times the amount of water to the dregs and decoct for another hour. Filter while hot. Combine the two filtrates and concentrate under reduced pressure at 40℃ to obtain an extract containing 1g of raw herbs per milliliter.

[0165] 2.2 Cell Culture

[0166] Same as Example 1.

[0167] 2.3 CCK-8 assay for cell viability

[0168] In this experiment, KGN cells were treated with different concentrations of composition 11 and formulations selected by uniform design method (0.1, 1, 10, 100 μg / mL) for 48 h to observe cell proliferation.

[0169] The specific steps are as follows:

[0170] (1) Take KGN cells in the logarithmic growth phase, prepare a single-cell suspension in DMEM / F12 medium containing 10% fetal bovine serum, and seed them into 96-well plates 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 received the same volume of culture medium as the drug groups. The normal control group was inoculated with cells and received the same volume of culture medium as the drug groups. After cell attachment, 10 μL of culture medium was added to the blank group and the normal control group. Each drug group received 10 μL of the corresponding 10-fold concentration of Composition 11 and the formulation selected using the uniform design method. Each group had three replicates. The final volume in each well was the same, 100 μL.

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

[0173] 2.4 ELISA method for detecting cellular E2 levels

[0174] In this experiment, KGN cells were treated with a 10 μg / mL formulation 11 and a uniform design method for 48 hours to detect cell E2 levels.

[0175] The specific steps are as follows:

[0176] (1) Take KGN cells in the logarithmic growth phase and prepare a single-cell suspension in DMEM / F12 medium without phenol red containing 10% fetal bovine serum. Seed the cells at a density of 5000 cells / well in a 96-well plate with a volume of 90 μL per well.

[0177] (2) After cell adhesion, 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 formulation screened by the uniform design method were added to each drug group. Three replicates were set for each group. The final volume in each well was the same, 100 μL; and

[0178] (3) 48 hours after 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 Example 1.

[0181] 3 Results

[0182] 3.1 The proliferative effect of composition 11 and the formulation screened by the uniform design method on KGN cells

[0183] Compared with the normal group, composition 11 (0.1, 1, 10, 100 μg / mL) and the formulations screened by the uniform design method (10, 100 μg / mL) significantly promoted KGN cell proliferation (Table 4). Figure 3 Composition 11 showed the best effect in promoting KGN cell proliferation.

[0184] Table 4. Effects of Composition 11 and Screened Formulations on KGN Cell Viability

[0185]

[0186] 3.2 Effects of Composition 11 and Formulations Screened Using the Uniform Design Method 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 selected by the uniform design method had no significant effect on the secretion of E2 in KGN cells.

[0188] 4. Conclusion

[0189] This invention utilizes human ovarian granulosa cells (KGN cells), currently the most commonly used cell type for studying changes in ovarian function. Apoptosis or non-apoptotic programmed cell death in granulosa cells is a major cause of follicular atresia; severe follicular atresia leads to follicular developmental disorders and premature ovarian aging. CCK-8 assay results show that the composition can promote KGN cell proliferation, thus exhibiting ovarian function protection and preventing ovarian dysfunction (DOR). Furthermore, multivariate regression analysis revealed that the optimal ratio of the composition for promoting KGN cell proliferation was 27:30:17:12:14 (Cuscuta chinensis: Lycium barbarum: Dioscorea opposita: Poria cocos: Nelumbo nucifera). To verify whether the selected formulation represents the optimal ratio, a comparative experiment was conducted between the composition 11, which showed the strongest effect in promoting KGN cell proliferation, and the formulation selected using a uniform design. The results of CCK8 and ELISA showed that composition 11 was more effective than the composition selected by uniform design. Therefore, it was concluded that the optimal formulation ratio of the composition was Cuscuta chinensis: Lycium barbarum: Dioscorea opposita: Poria cocos: Nelumbo nucifera = 4:4:2:1:1.

[0190] The embodiments of the present invention have been described in detail above. Specific examples have been used to illustrate the principles and implementation methods of the present invention. The descriptions of the embodiments above are only for the purpose of helping to understand the method and core ideas of the present invention. Furthermore, any changes or modifications made by those skilled in the art based on the ideas of the present invention, its specific implementation methods, and its application scope, are all within the scope of protection of the present invention. Therefore, the content of this specification should not be construed as a limitation of the present invention.

Claims

1. The use of a pharmaceutical composition comprising Cuscuta chinensis in the preparation of a medicament for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency, and / or premature ovarian failure, characterized in that, The active pharmaceutical ingredient of the pharmaceutical composition, by weight, comprises 48.6-59.4 parts of Cuscuta chinensis, 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 Nelumbo nucifera; or 54-66 parts of Cuscuta chinensis, 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 Nelumbo nucifera; or 24.3-29.7 parts of Cuscuta chinensis, 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 Nelumbo nucifera.

2. The use of a pharmaceutical composition comprising Cuscuta chinensis in the preparation of a medicament for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency, and / or premature ovarian failure, characterized in that, The active pharmaceutical ingredient of the pharmaceutical composition is composed of Cuscuta chinensis, Lycium barbarum, Dioscorea opposita, Poria cocos and Nelumbo nucifera, wherein the weight ratio of Cuscuta chinensis, Lycium barbarum, Dioscorea opposita, Poria cocos and Nelumbo nucifera is (3.6~4.4):(3.6~4.4):(1.8~2.2):(0.9~1.1):(0.9~1.1).

3. The use according to claim 1 or 2, characterized in that, The pharmaceutical composition is a traditional Chinese medicine composition.

4. The use of a pharmaceutical preparation containing Cuscuta chinensis in the preparation of a medicament for the prevention and / or treatment of decreased ovarian reserve, premature ovarian insufficiency, and / or premature ovarian failure, characterized in that, The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1 to 3 and a pharmaceutically acceptable excipient.

5. The use according to claim 4, characterized in that, The excipients are selected from one or more of the following: diluents, wetting agents, adhesives, disintegrants, encapsulating agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers, and antioxidants.

6. The use according to claim 4, characterized in that, The pharmaceutical preparations are pills, powders, tablets, drop pills, capsules, films, lozenges, granules, injections, or oral liquids.

7. The use according to claim 6, characterized in that, The pharmaceutical preparation is in the form of pills.

8. The use according to claim 1 or 2, characterized in that, The preparation method of the pharmaceutical composition includes the following steps: (1) Weigh out appropriate amounts of dodder seed, wolfberry fruit, yam, poria cocos and lotus seed, mix them evenly to obtain a mixture, add water to decoct, filter to obtain the first filtrate and dregs; (2) Boil the dregs in water, filter, and obtain a second filtrate; as well as (3) Combine the first filtrate and the second filtrate, concentrate them, and obtain the drug composition.

9. The use according to claim 8, characterized in that, In step (1), the volume / mass ratio of the water to the mixture is 8 to 12.

10. The use according to claim 9, characterized in that, The volume / mass ratio of the water to the mixture is 10.

11. The use according to claim 8, characterized in that, In step (1), the filtration is hot filtration.

12. The use according to claim 8, characterized in that, In step (1), the boiling time with water is 0.5 to 2 hours.

13. The use according to claim 12, characterized in that, The boiling time with water is 0.9~1.1 h.

14. The use according to claim 8, characterized in that, In step (2), the volume / mass ratio of the water to the dregs is 8 to 12 times.

15. The use according to claim 14, characterized in that, The volume / mass ratio of the water to the medicinal residue is 10.

16. The use according to claim 8, characterized in that, In step (2), the filtration is hot filtration.

17. The use according to claim 8, characterized in that, In step (2), the boiling time with water is 0.5 to 2 hours.

18. The use according to claim 17, characterized in that, The boiling time with water is 0.9~1.1 h.

19. The use according to claim 8, characterized in that, In step (3), the concentration is reduced pressure concentration.

20. The use according to claim 8, characterized in that, In step (3), the concentration temperature is 30℃~60℃.

21. The use according to claim 20, characterized in that, The concentration temperature is 40°C.

22. The use according to claim 8, characterized in that, The concentration of the pharmaceutical composition is 0.5-2g of crude drug per milliliter.

23. The use according to claim 22, characterized in that, The concentration of the pharmaceutical composition is 0.9-1.1 g of crude drug per milliliter.

24. The use according to claim 1 or 2, characterized in that, The decline in ovarian reserve function is referred to as pathological decline in ovarian reserve function.

25. The use according to claim 1 or 2, characterized in that, The prevention and / or treatment of decreased ovarian reserve function involves promoting the proliferation of human ovarian granulosa cells.

26. The use according to claim 25, characterized in that, The human ovarian granulosa cells mentioned are KGN cells.

27. The use according to claim 2, characterized in that, The decline in ovarian reserve function is defined as a decrease in ovarian estradiol levels.

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