A traditional Chinese medicine composition for treating premature ovarian failure, a preparation method and application thereof

Abstract

The application belongs to the technical field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating early ovarian dysfunction and a preparation method and application thereof. The traditional Chinese medicine composition is composed of the following raw materials in parts by weight: Radix Rehmanniae Preparata 2-12 parts, Herba Rhizomatis 2-8 parts, Herba Epimedii 1-6 parts, Semen Euryale 2-12 parts, Angelica sinensis 1-6 parts, Cyperus rotundus 1-6 parts, Radix Paeoniae Alba 2-6 parts, Eucommia ulmoides 2-6 parts, Phellodendron amurense 1-6 parts and Rhizoma Anemarrhenae 2-6 parts. The traditional Chinese medicine composition is rigorous in prescription, has the functions of nourishing kidney and warming yang, soothing liver and relieving depression, and clearing and eliminating deficiency heat, the raw material components are cheap and easy to obtain, the whole prescription has the functions of supplementing yin and yang, treating liver and kidney simultaneously, and applying supplementation and purgation simultaneously, can effectively regulate the endocrine level of the patient, reduce the serum follicle stimulating hormone (FSH) level, and increase the basic antral follicle count.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a traditional Chinese medicine composition for treating early-onset ovarian insufficiency, its preparation method, and its application. Background Technology

[0002] Premature ovarian insufficiency (POI) is a clinical syndrome characterized by decreased ovarian function in women before the age of 40. Clinical manifestations include menstrual abnormalities (amenorrhea, oligomenorrhea, or polymenorrhea), elevated gonadotropin levels, and fluctuating estrogen levels, often accompanied by infertility and hypoestrosis-related symptoms (such as hot flashes, sweating, sleep disturbances, and mood swings). POI exhibits high heterogeneity, progresses rapidly, and is often irreversible. Without timely intervention, it can further develop into premature ovarian failure (POF). Furthermore, POI patients have a significantly increased risk of long-term osteoporosis, cardiovascular disease, cognitive impairment, and metabolic disorders, severely impacting women's reproductive health and overall quality of life.

[0003] The etiology of poikiloma (POI) is complex and diverse, involving genetic factors (including chromosomal abnormalities and gene mutations), immune factors (such as autoimmune diseases and anti-thyroid autoantibodies), iatrogenic factors (such as radiation therapy, chemotherapy drugs, and ovarian surgery), and environmental factors. However, most POI patients have idiopathic POI, and its exact pathogenesis remains unclear. Studies have shown that the pathogenesis of POI may involve the interaction between genes and environmental factors. Recent whole-exome sequencing and genome-wide association studies have revealed the complex genetic structure of POI, including monogenic and oligogenic inheritance patterns, which poses significant challenges to genetic diagnosis.

[0004] In terms of Western medicine treatment, current clinical treatment options for POI (Potential Ovarian Insufficiency) remain limited both domestically and internationally, with no interventions capable of reversing ovarian function or curing POI. Hormone replacement therapy (HRT) is currently the main Western medical treatment for POI. It simulates the menstrual cycle through sequential estrogen and progesterone therapy, aiming to improve symptoms of low estrogen (such as hot flashes, night sweats, and vaginal dryness), reduce the risk of cardiovascular disease and osteoporosis, and continue hormone supplementation at least until normal menopause age. However, HRT cannot restore ovarian reserve and ovulation function, has a high relapse rate after discontinuation, and long-term use carries potential risks of endometrial cancer, breast cancer, and thrombosis. Some patients refuse or discontinue treatment due to safety concerns. Assisted reproductive technologies such as in-vitro fertilization (IVF) are difficult to use as a routine and effective method in POI patients due to severely insufficient ovarian reserve, high cycle cancellation rates, and extremely low live birth rates. While egg or embryo donation can solve fertility problems, it faces practical difficulties such as ethical controversies, donor scarcity, and high costs. Compared to Western medicine treatments, the advantage of Traditional Chinese Medicine (TCM) lies in its holistic regulation of the "Kidney-Tian Gui-Chong Ren-Uterus" axis. Through methods such as tonifying the kidney and replenishing essence, soothing the liver and nourishing blood, and promoting blood circulation and removing blood stasis, it can fundamentally improve ovarian function, promote follicle development, and enhance the body's immunity and antioxidant capacity. Moreover, it has fewer adverse reactions and higher patient acceptance. However, there is still a lack of commercially available TCM preparations with definite efficacy and simplified components.

[0005] Chinese invention patent application CN110755536A discloses a traditional Chinese medicine composition, the components of which include Rehmannia glutinosa, Epimedium brevicornu, Paeonia lactiflora, Lycium barbarum, Testudinis plastrum, Anemarrhena asphodeloides, Cuscuta chinensis, Morinda officinalis, Cistanche deserticola, Phellodendron chinense, Coptis chinensis, Poria cocos, Bupleurum chinense, Angelica sinensis, and Ligusticum chuanxiong, etc. It has the effects of improving ovarian reserve function, increasing the number of primordial follicles and primary follicles, and downregulating serum follicle-stimulating hormone, and can be used to treat diseases related to ovarian reserve deficiency caused by chronic stress.

[0006] Chinese invention patent application CN109528980A discloses a traditional Chinese medicine composition for treating diseases of decreased ovarian reserve. It is made from raw materials such as Rehmannia glutinosa, Polygonatum sibiricum, Taxillus chinensis, Dipsacus asper, Ligustrum lucidum, Lycium barbarum, Salvia miltiorrhiza, and Curcuma longa. It has the effects of nourishing the kidney and replenishing essence, nourishing blood and soothing the liver, and nourishing yin and raising yang. It is mainly used to treat kidney essence deficiency and liver stagnation and blood deficiency syndrome. It can be used to treat diseases such as decreased ovarian reserve, premature ovarian insufficiency, and premature ovarian failure.

[0007] Chinese invention patent application CN110179947A discloses a Yin-nourishing and liver-soothing drug, the effective components of which are composed of the following raw materials in parts by weight: 10-30 parts Rehmannia glutinosa, 10-20 parts Paeonia lactiflora, 5-15 parts Bupleurum chinense, 10-15 parts Curcuma longa, 5-15 parts Cyperus rotundus, 10-30 parts Dioscorea opposita, and 10-15 parts Paeonia lactiflora. It can effectively regulate the patient's basal endocrine level, increase the number of basal antral follicles, and improve ovarian blood flow, with significant therapeutic effects. It can effectively improve menstrual disorders, ovarian function, and symptoms of kidney deficiency and liver stagnation in patients with ovarian insufficiency.

[0008] However, the above-mentioned traditional Chinese medicine compositions still have the following shortcomings in treating POI: First, some compositions have a large number of components, such as CN110755536A which contains as many as 15 raw materials and CN117442659A which contains 14 raw materials, increasing the cost of preparation and production difficulty, which is not conducive to long-term use by patients and industrial promotion; Second, existing studies have mostly focused on improving serum hormone levels and baseline antral follicle count, and there is still insufficient evidence of efficacy in restoring natural ovulation function and improving pregnancy rate.

[0009] Therefore, there is an urgent need in this field for a traditional Chinese medicine composition that is rationally formulated, has definite curative effect, can comprehensively cover the complex pathogenesis of POI, and has high safety. Summary of the Invention

[0010] The purpose of this invention is to provide a traditional Chinese medicine composition for treating early-onset ovarian insufficiency, and related technologies, to solve the technical problems of existing compositions such as numerous components, unclear efficacy, and low safety, or combinations thereof.

[0011] Terminology Explanation: Unless otherwise defined, all technical terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this subject matter pertains. Unless otherwise stated, all patents, patent inventions, and disclosures cited throughout this document are incorporated herein by reference in their entirety. Where multiple definitions exist for terms herein, the definitions provided in this chapter shall prevail.

[0012] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0013] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0014] The terms “optional / arbitrary” or “optionally / arbitrarily” mean that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0015] The term "prepared Rehmannia root" used in this article refers to: raw Rehmannia root (a plant of the Scrophulariaceae family). Rehmannia sticky Processed product of fresh or dried tubers of Libosch. It has a sweet taste and slightly warm nature. It enters the liver and kidney meridians. Functions and indications: Nourishes blood and yin, replenishes essence and marrow. Used for blood deficiency and chlorosis, palpitations, irregular menstruation, metrorrhagia, liver and kidney yin deficiency, soreness and weakness of the lower back and knees, bone steaming fever, night sweats and seminal emission, internal heat and thirst, dizziness, tinnitus, and premature graying of hair.

[0016] Preparation of Rehmannia glutinosa: Take raw Rehmannia glutinosa and steam it according to the steaming method (Pharmacopoeia General Chapter 0213) until it turns black and moist. Take it out and sun-dry it until it is about 80% dry. Cut it into thick slices or pieces and dry it.

[0017] The term "Xianmao" as used in this article refers to the plant *Curculigo orchioides*, belonging to the Amaryllidaceae family. Orchid Warbler The dried rhizome of Gaertn. It has a pungent taste and is hot in nature; it is poisonous. It enters the kidney, liver, and spleen meridians. Its functions and indications include: tonifying kidney yang, strengthening tendons and bones, and dispelling cold and dampness. It is used for impotence, premature ejaculation, weakness of tendons and bones, cold pain in the lower back and knees, and diarrhea due to yang deficiency.

[0018] The term "Epimedium" used in this article refers to Epimedium, a plant belonging to the Berberidaceae family. Epimedium brevicornum Maxim., Epimedium sarcodactylis Epimedium sagittatum (Sieb. et Zucc.) Maxim., Epimedium brevicornu Epimedium pubescent Maxim. or Korean Epimedium Epimedium koreanum Dried leaves of Nakai. Taste: pungent, sweet; nature: warm. Meridians: liver, kidney. Functions and indications: tonifies kidney yang, strengthens tendons and bones, dispels wind and dampness. Used for kidney yang deficiency, impotence, seminal emission, weakness of tendons and bones, rheumatic pain, numbness, and contractures.

[0019] The term "dodder" used in this article refers to Cuscuta chinensis, a plant belonging to the Convolvulaceae family. Southern Dodder R. Br. or dodder Chinese DodderDried, mature seeds of Lam. Taste: pungent and sweet; nature: neutral. Meridians: liver, kidney, spleen. Functions and indications: Tonifies the liver and kidneys, strengthens essence and reduces urination, calms the fetus, improves eyesight, stops diarrhea; externally used to dispel wind and remove blemishes. Used for liver and kidney deficiency, soreness and weakness of the lower back and knees, impotence, seminal emission, enuresis, frequent urination, kidney deficiency with threatened abortion, threatened miscarriage, blurred vision and tinnitus, spleen and kidney deficiency with diarrhea; externally used to treat vitiligo.

[0020] The term "Angelica sinensis" as used in this article refers to Angelica sinensis, a plant belonging to the Apiaceae family. Angelica sinensis The dried root of *Oliv.* Diels. Sweet and pungent in taste, warm in nature. It enters the liver, heart, and spleen meridians. Functions and indications: Nourishes and invigorates blood, regulates menstruation and relieves pain, moistens the intestines and promotes bowel movement. Used for blood deficiency and chlorosis, dizziness and palpitations, irregular menstruation, amenorrhea and dysmenorrhea, abdominal pain due to deficiency and cold, rheumatic arthralgia, traumatic injuries, carbuncles and sores, and constipation due to intestinal dryness. Wine-processed *Angelica sinensis* invigorates blood and regulates menstruation. Used for amenorrhea and dysmenorrhea, rheumatic arthralgia, and traumatic injuries.

[0021] The term "Cyperus rotundus" used in this article refers to Cyperus rotundus, a plant belonging to the Cyperaceae family. Rounded bulrush The dried rhizome of L. Tastes pungent, slightly bitter, and slightly sweet; neutral in nature. Enters the liver, spleen, and triple burner meridians. Functions and indications: Soothes the liver and relieves stagnation, regulates qi and relieves chest tightness, regulates menstruation and relieves pain. Used for liver qi stagnation, chest and rib pain, hernia pain, breast pain, spleen and stomach qi stagnation, abdominal distension and pain, irregular menstruation, amenorrhea, and dysmenorrhea.

[0022] The term "white peony" as used in this article refers to the peony plant (Paeonia lactiflora), a member of the Ranunculaceae family. Paeonia lactiflora The dried root of Pall. It tastes bitter and sour, and is slightly cold in nature. It enters the liver and spleen meridians. Its functions and indications include nourishing blood and regulating menstruation, astringing yin and stopping sweating, softening the liver and relieving pain, and calming liver yang. It is used for blood deficiency and chlorosis, irregular menstruation, spontaneous sweating, night sweats, hypochondriac pain, abdominal pain, limb spasms, headache, and dizziness.

[0023] The term "Eucommia" as used in this article refers to Eucommia ulmoides, a plant belonging to the Eucommiaceae family. Eucommia elmoides Dried bark of Oliv. Sweet in taste and warm in nature. It enters the liver and kidney meridians. Functions and indications: Tonifies the liver and kidneys, strengthens tendons and bones, and calms the fetus. Used for liver and kidney deficiency, lower back and knee pain, weakness of tendons and bones, dizziness, threatened miscarriage, and fetal restlessness.

[0024] The term "Huangbai" used in this article refers to the yellow bark tree (Phellodendron amurense), a plant belonging to the Rutaceae family. Phellodendron in Chinese Dried bark of Schneid. Bitter in taste and cold in nature. It enters the kidney and bladder meridians. Functions and indications: Clears heat and dries dampness, drains fire and eliminates steaming heat, detoxifies and heals sores. Used for damp-heat diarrhea, jaundice with dark urine, leukorrhea and vulvar itching, painful urination due to heat, beriberi with weakness and paralysis, steaming bone fever, night sweats, seminal emission, sores and carbuncles, eczema and damp sores. Salt-processed Phellodendron bark nourishes yin and reduces fire. Used for yin deficiency with excessive fire, night sweats and steaming bone fever.

[0025] The term "Anemarrhena asphodeloides" used in this article refers to the plant Anemarrhena asphodeloides, belonging to the Liliaceae family. Anemarrhena asphodeloidesThe dried rhizome of Bge. It tastes bitter and sweet, and is cold in nature. It enters the lung, stomach, and kidney meridians. Functions and indications: Clears heat and drains fire, nourishes yin and moistens dryness. Used for exogenous febrile diseases, high fever and thirst, dry cough due to lung heat, steaming bone fever, internal heat and thirst, and constipation due to intestinal dryness.

[0026] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating early-onset ovarian insufficiency, the traditional Chinese medicine composition comprising the following raw materials in parts by weight: 2-12 parts of Rehmannia glutinosa, 2-8 parts of Curculigo orchioides, 1-6 parts of Epimedium brevicornu, 2-12 parts of Cuscuta chinensis, 1-6 parts of Angelica sinensis, 1-6 parts of Cyperus rotundus, 2-6 parts of Paeonia lactiflora, 2-6 parts of Eucommia ulmoides, 1-6 parts of Phellodendron chinense, and 2-6 parts of Anemarrhena asphodeloides.

[0027] Preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 6-10 parts of Rehmannia glutinosa, 3-5 parts of Curculigo orchioides, 3-5 parts of Epimedium brevicornu, 6-10 parts of Cuscuta chinensis, 3-5 parts of Angelica sinensis, 3-5 parts of Cyperus rotundus, 3-5 parts of Paeonia lactiflora, 3-5 parts of Eucommia ulmoides, 3-5 parts of Phellodendron chinense, and 3-5 parts of Anemarrhena asphodeloides.

[0028] More preferably, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 8 parts Rehmannia glutinosa, 4 parts Curculigo orchioides, 4 parts Epimedium brevicornu, 8 parts Cuscuta chinensis, 4 parts Angelica sinensis, 4 parts Cyperus rotundus, 4 parts Paeonia lactiflora, 4 parts Eucommia ulmoides, 4 parts Phellodendron chinense, and 4 parts Anemarrhena asphodeloides.

[0029] Secondly, the present invention provides a method for preparing the above-mentioned traditional Chinese medicine composition, comprising the following steps: Mix all the ingredients, add water and boil, filter, combine the filtrates, concentrate, dry, and pulverize to obtain the final product.

[0030] The conditions for adding water and decocting are as follows: the number of decoctions is 1-3, the amount of water added each time is 8-10 times the total mass of the medicinal materials, and the decoction time each time is 0.5-1.5 hours.

[0031] The concentration conditions are: temperature 60℃-75℃, vacuum degree -0.08 MPa to -0.10 MPa; the drying conditions are: temperature ≤70℃, vacuum degree -0.04 MPa to -0.10 MPa.

[0032] Thirdly, the present invention provides a pharmaceutical preparation comprising the above-described traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the above-described preparation method, and pharmaceutically acceptable excipients.

[0033] The dosage form of the pharmaceutical preparation is selected from granules, powders, decoctions, pills, tablets, capsules, mixtures, or syrups.

[0034] Wherein, the pharmaceutically acceptable excipients are selected from one or more of fillers, disintegrants, glidants, binders, flavoring agents, emulsifiers, suspending agents, sustained-release materials, propellants, dispersants, and coating materials.

[0035] Fourthly, the present invention provides the use of the above-mentioned traditional Chinese medicine composition or the traditional Chinese medicine composition or the pharmaceutical preparation prepared by the above-mentioned preparation method in the preparation of a drug for treating early-onset ovarian insufficiency.

[0036] In this formula, Rehmannia glutinosa nourishes Yin and blood, and regulates and tonifies the liver and kidneys; Curculigo orchioides warms the kidneys and assists Yang, tonifying the fire of the gate of life. These two herbs complement each other, working together to nourish the kidneys, warm Yang, and harmonize the Chong and Ren meridians, serving as the principal herbs. Epimedium strengthens Yang, assists fire, and fortifies tendons and bones, enhancing the warming effect of Curculigo orchioides on kidney Yang; Cuscuta chinensis tonifies Yin and Yang, assisting the principal herbs in harmonizing Yin and Yang within the kidneys; Angelica sinensis tonifies essence and blood, regulates menstruation, and relieves pain, assisting Rehmannia glutinosa in replenishing the blood sea and nourishing the uterus; Eucommia ulmoides tonifies the liver and kidneys, strengthens tendons and bones, assisting the principal herbs in protecting the kidneys. These four herbs work together as assistant herbs to enhance the effects of tonifying the kidneys, replenishing essence, and regulating the Chong and Ren meridians. Paeonia lactiflora nourishes blood, softens the liver, astringes Yin, and relieves pain, both moistening the liver and counteracting the detrimental effects of warming and drying herbs on Yin; Cyperus rotundus soothes the liver, relieves depression, regulates Qi, and regulates menstruation, both soothing liver Qi and preventing tonifying and nourishing herbs from obstructing Qi flow, ensuring the entire formula is tonifying without stagnation and promoting the smooth flow of Qi and blood. One is gentle, the other gentle, serving as adjuvant herbs. Phellodendron bark clears heat and dries dampness, reducing deficiency heat, specifically targeting the lower burner, effectively clearing deficiency fire in the kidneys; Anemarrhena rhizome clears heat and drains fire, nourishes yin and moistens dryness, clearing internal deficiency heat while also nourishing yin and generating fluids. These two herbs work synergistically, guiding the other herbs in the formula directly to the lower burner and kidneys, serving as guiding herbs. The entire formula tonifies both yin and yang, treats both the liver and kidneys, and combines tonification and purgation, replenishing kidney essence, harmonizing yin and yang, soothing liver qi, and clearing deficiency heat. This leads to abundant Tian Gui (menstrual blood), smooth flow of the Chong and Ren meridians, and proper storage and release in the uterus, restoring ovarian function, normalizing menstruation, and increasing the chances of conception. It regulates the function of the "kidney-Tian Gui-Chong and Ren-uterus" reproductive axis, improves ovarian reserve, promotes follicle development and ovulation, restores the menstrual cycle, and enhances fertility. It is suitable for early-onset ovarian insufficiency (POI) diagnosed in Traditional Chinese Medicine as kidney deficiency and liver stagnation, yin and yang deficiency, and concurrent deficiency heat.

[0037] Compared with the prior art, the present invention has the following beneficial effects: The traditional Chinese medicine composition of this invention is meticulously formulated and possesses the functions of nourishing the kidneys and warming yang, soothing the liver and relieving depression, and clearing away deficiency heat. Its raw materials are inexpensive and readily available. The entire formula tonifies both yin and yang, treats both the liver and kidneys, and combines tonification and purgation, effectively regulating the patient's endocrine levels, lowering FSH, and increasing the baseline antral follicle count. It is significantly effective for premature ovarian insufficiency due to kidney deficiency and liver stagnation, yin and yang deficiency, and concurrent deficiency heat. It can effectively improve menstrual disorders, restore ovarian function, and relieve clinical symptoms such as hot flashes, night sweats, lower back and knee weakness, and irritability. Furthermore, it has few side effects, high patient compliance with long-term use, and good safety, thus overcoming the shortcomings of existing hormone replacement therapy that cannot restore ovarian function and carries long-term safety risks. It has broad clinical application prospects and promotional value. Detailed Implementation

[0038] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0039] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0040] Example 1 A traditional Chinese medicine composition for treating early-onset ovarian insufficiency (POI) comprises the following ingredients by weight: 8 parts Rehmannia glutinosa, 4 parts Curculigo orchioides, 4 parts Epimedium brevicornu, 8 parts Cuscuta chinensis, 4 parts Angelica sinensis, 4 parts Cyperus rotundus, 4 parts Paeonia lactiflora, 4 parts Eucommia ulmoides, 4 parts Phellodendron chinense, and 4 parts Anemarrhena asphodeloides.

[0041] The preparation method is as follows: (1) Take all the raw materials and mix them together. Add water and decoct three times. For the first time, add 10 times the total weight of the medicinal materials and decoct for 1 hour. Filter (100 mesh). For the second time, add 8 times the total weight of the medicinal materials and decoct for 1 hour. Filter (100 mesh). For the third time, add 8 times the total weight of the medicinal materials and decoct for 1 hour. Filter (100 mesh). Combine the filtrates.

[0042] (2) The filtrate was concentrated at 65℃ and vacuum degree -0.09 MPa, dried (65℃, -0.06 MPa), pulverized, and passed through an 80-mesh sieve to obtain dry paste powder.

[0043] Example 2 A traditional Chinese medicine composition for treating early-onset ovarian insufficiency (POI) comprises the following ingredients by weight: 2 parts Rehmannia glutinosa, 8 parts Curculigo orchioides, 2 parts Epimedium brevicornu, 12 parts Cuscuta chinensis, 2 parts Angelica sinensis, 2 parts Cyperus rotundus, 2 parts Paeonia lactiflora, 2 parts Eucommia ulmoides, 2 parts Phellodendron chinense, and 2 parts Anemarrhena asphodeloides.

[0044] The preparation method is the same as in Example 1.

[0045] Example 3 A traditional Chinese medicine composition for treating early-onset ovarian insufficiency (POI) comprises the following ingredients by weight: 2 parts Rehmannia glutinosa, 2 parts Curculigo orchioides, 6 parts Epimedium brevicornu, 2 parts Cuscuta chinensis, 6 parts Angelica sinensis, 6 parts Cyperus rotundus, 2 parts Paeonia lactiflora, 2 parts Eucommia ulmoides, 6 parts Phellodendron chinense, and 2 parts Anemarrhena asphodeloides.

[0046] The preparation method is the same as in Example 1.

[0047] Example 4 A traditional Chinese medicine composition for treating early-onset ovarian insufficiency (POI) comprises the following ingredients by weight: 10 parts Rehmannia glutinosa, 2 parts Curculigo orchioides, 1 part Epimedium brevicornu, 2 parts Cuscuta chinensis, 1 part Angelica sinensis, 1 part Cyperus rotundus, 6 parts Paeonia lactiflora, 6 parts Eucommia ulmoides, 1 part Phellodendron chinense, and 6 parts Anemarrhena asphodeloides.

[0048] The preparation method is the same as in Example 1.

[0049] Comparative Example 1 The only difference between this comparative example and Example 1 is that Curculigo orchioides is replaced with Morinda officinalis.

[0050] Comparative Example 2 The only difference between this comparative example and Example 1 is that Cuscuta chinensis is replaced with Lycium barbarum.

[0051] Comparative Example 3 The only difference between this comparative example and Example 1 is that Anemarrhena asphodeloides is replaced with Eclipta prostrata.

[0052] Comparative Example 4 This comparative example uses the composition in the existing technology CN108324821A. By weight, the raw materials are: 3 parts of Ligustrum lucidum, 3 parts of Eclipta prostrata, 3 parts of Lycium barbarum, 3 parts of Cuscuta chinensis, 2 parts of Paeonia lactiflora, 2 parts of Rehmannia glutinosa, 2 parts of Ligusticum chuanxiong, 2 parts of Cyperus rotundus (processed with vinegar), and 1 part of Glycyrrhiza uralensis (processed with vinegar).

[0053] The preparation method is the same as in Example 1.

[0054] Effect test 1. Experimental Model Six female SD rats were randomly selected as the normal control group, and the rest were used for modeling. The model animals were injected intraperitoneally with vinylcyclohexene dioxide (VCD) at a dose of 160 mg / kg (injection site disinfected) to induce ovarian dysfunction. The injection was repeated for 15 days. Vaginal smears were performed starting on the 8th day of modeling, once a day from 9:00 to 10:00, for 8 consecutive days. Rats with disordered or prolonged estrous cycles and good physical condition were selected for the experiment.

[0055] VCD induction model dosage calculation: Vinylcyclohexene dioxide (VCD) was injected intraperitoneally at a dose of 160 mg / kg. The density of VCD was 1.094 g / mL, and the administration volume was 0.146 mL / kg. The VCD stock solution was diluted with corn oil at a ratio of 1:16 to prepare the modeling solution, which was then injected intraperitoneally at a dose of 2.482 mL / kg.

[0056] 2. Grouping and administration After modeling was completed, the successfully modeled rats were divided into 10 groups: model control group, estradiol valerate tablet group, Example 1 group, Example 2 group, Example 3 group, Example 4 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group, with 6 rats in each group. Each group was administered the corresponding dry extract powder prepared in the example, once daily by gavage at a dose of 4.2 g / kg / day (based on dry extract powder), for 5 consecutive weeks. The normal control group and the model control group were given an equal volume of pure water.

[0057] 3. Detection indicators and methods During the experiment, the general condition, estrous cycle, and weight changes of rats were observed. Serum follicle-stimulating hormone (FSH) was measured before administration and 5 weeks later. Five weeks after administration, the animals were euthanized under anesthesia, and the wet weight of the ovaries was measured to calculate the ovarian coefficient (ovarian coefficient (%) = ovarian weight (mg) / body weight (g) × 100%). Ovarian tissue pathological examination and follicle counting were performed, and the relative expression level of the ovarian apoptosis-related gene Bcl-2 was measured.

[0058] The relative expression level of the apoptosis-related gene Bcl-2 in ovarian tissue was detected by sacrificing rats, collecting ovarian tissue, rinsing the surface with sterile 0.9% sodium chloride solution to remove blood, drying the tissue, placing it in pre-cooled nuclease-free cryovials, flash-freezing in liquid nitrogen, and storing it at -80°C for later analysis. The content of the B-cell lymphoma-2 gene (Bcl-2) in ovarian tissue was detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). RT-qPCR was used to detect the expression level of B-cell lymphoma-2 in ovarian tissue. -ΔΔCt The relative expression levels of each gene group are calculated, and the specific calculation steps are as follows: ① Calculate ΔCt for all samples ΔCt(sample) = Ct(Bcl-2) - Ct(internal reference gene β-actin); ② Calculate ΔCt ΔΔCt = ΔCt (other groups) - ΔCt (model control group). The model control group is the baseline, so ΔΔCt = 0 for the model control group. Note: In this experiment, the "model control group" was used as the control, and all other groups were compared with the model control group.

[0059] ③ Calculate the change factor Relative expression level = 2 -ΔΔCt That is, you get it.

[0060] 4. Test Results (1) General state observation During the experiment, no abnormal signs were observed in the animals in the normal control group.

[0061] From the first week after administration of the modeling agent, the animals exhibited piloerection, arched back posture, abdominal distension, drooling (mild to moderate) and / or emaciation, dark red discharge around the eyes / nose, prone position, and reduced activity. These signs persisted during the administration period and gradually recovered from day 21 (the first week after discontinuation of the modeling agent).

[0062] (2) Follicle-stimulating hormone At the end of the modeling process (before drug administration), the follicle-stimulating hormone (FSH) levels in the modeling rats were significantly elevated (compared to the normal control group). P The value <0.01 indicates that the ovarian function impairment model was successfully established.

[0063] During week 5 of drug administration, compared with the model control group, all drug administration groups showed a decreasing trend in follicle-stimulating hormone (FSH). Among them, the decrease in groups 1 and 2 was statistically significant. P <0.05 or P The difference was <0.01, indicating that Groups 1 and 2 of Examples could significantly reverse the abnormal increase in serum follicle-stimulating hormone (FSH) in rats with ovarian dysfunction; Comparative Groups 1 and 2 also showed a decreasing trend, but the difference did not reach statistical significance.

[0064] The results are shown in Table 1 below.

[0065] Table 1. Results of serum follicle-stimulating hormone (FSH) changes.

[0066] Note: Compared with the normal control group, P <0.01; compared with the model control group, & P <0.05, && P <0.01; compared with Example 1 group, # P <0.05.

[0067] (3) Relative expression level of Bcl-2 gene in ovarian tissue At week 5 of drug administration, there was a significant difference between the model group and the drug administration group, indicating that the ovarian function impairment model was maintained effectively.

[0068] Compared with the model control group, the relative expression level of Bcl-2 gene in each drug-treated group showed an increasing trend. Among them, the increase in groups 1 and 3 was statistically significant. P <0.05 or P <0.01), indicating that Groups 1 and 3 of Examples can significantly restore the decreased Bcl-2 expression in the ovarian tissue of rats caused by ovarian dysfunction; Comparative Groups 3 and 4 also showed an increasing trend.

[0069] The results are shown in Table 2 below.

[0070] Table 2. Results of relative expression levels of the Bcl-2 gene in ovarian tissue.

[0071] Note: Compared with the model control group, & P <0.05, && P <0.01; compared with Example 1 group, # P <0.05.

[0072] (4) Ovarian weight and coefficient At week 5 of drug administration, there was a significant difference between the model group and the drug administration group, indicating that the ovarian function impairment model was maintained effectively.

[0073] Compared with the model control group, the ovarian coefficients of all treatment groups showed an increasing trend. Among them, the increases in Group 1 and Group 2 were statistically significant. P The value <0.01 indicates that groups 1 and 2 of Examples significantly increased the ovarian coefficient of rats caused by ovarian dysfunction; comparative groups 3 and 4 also showed an increasing trend.

[0074] The results are shown in Table 3 below.

[0075] Table 3 Results of rat ovarian coefficient test

[0076] Note: Compared with the model control group, && P<0.05; compared with Example 1 group, # P <0.05, ## P <0.01.

[0077] (5) Follicle count After 5 weeks of drug administration, the number of secondary follicles in the model control group was significantly lower than that in the normal control group. P <0.01). Compared with the model control group, the number of secondary follicles in each group of the test sample showed an increasing trend; among them, the increase in Example 1 group and Example 2 group was statistically significant; Comparative Example 3 group and Comparative Example 4 group were statistically significant compared with Example 1 group. See Table 4 below for details.

[0078] Table 4. Results of follicle count detection

[0079] Note: Compared with the model control group, & P<0.05, && P <0.01; compared with Example 1 group, # P <0.05.

[0080] (6) Pathological examination Gross pathological examination results: In the normal control group, the overall organ structure of rats was intact, the anatomical layers of the abdominal organs were clear, and no obvious abnormalities were found. Except for the normal control group, some animals in the other groups showed adhesion of the abdominal organs, mainly manifested as adhesion of the liver, diaphragm, stomach, spleen, intestines, and ovaries, and / or accompanied by red ascites, ranging from about 10-60 mL.

[0081] Histopathological examination results: Except for the normal control group, a few animals in the model control group, estradiol valerate tablet group, Examples 2-4 groups and Comparative Examples 1-4 groups had unilateral ovarian follicular cysts. Only Example 1 group did not have ovarian follicular cysts, indicating that the composition of Example 1 may have a certain protective effect against follicular cysts.

[0082] Follicles: In the normal control group, rats had an ample reserve and abundant number of primordial follicles in their ovaries, with complete developmental stages of primary and secondary follicles. Compared with the normal control group, the number of secondary follicles in the model control group was reduced.

[0083] The experimental results of the above groups show that the treatment regimens in Examples 1-4 have a certain therapeutic effect on rats with ovarian dysfunction, and that replacing the ingredients or using other regimens cannot achieve the original therapeutic effect.

[0084] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A traditional Chinese medicine composition for treating early-onset ovarian insufficiency, characterized in that, The traditional Chinese medicine composition consists of the following raw materials in parts by weight: 2-12 parts of Rehmannia glutinosa, 2-8 parts of Curculigo orchioides, 1-6 parts of Epimedium brevicornu, 2-12 parts of Cuscuta chinensis, 1-6 parts of Angelica sinensis, 1-6 parts of Cyperus rotundus, 2-6 parts of Paeonia lactiflora, 2-6 parts of Eucommia ulmoides, 1-6 parts of Phellodendron chinense, and 2-6 parts of Anemarrhena asphodeloides.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The traditional Chinese medicine composition consists of the following raw materials in parts by weight: 6-10 parts of Rehmannia glutinosa, 3-5 parts of Curculigo orchioides, 3-5 parts of Epimedium brevicornu, 6-10 parts of Cuscuta chinensis, 3-5 parts of Angelica sinensis, 3-5 parts of Cyperus rotundus, 3-5 parts of Paeonia lactiflora, 3-5 parts of Eucommia ulmoides, 3-5 parts of Phellodendron chinense, and 3-5 parts of Anemarrhena asphodeloides.

3. The traditional Chinese medicine composition according to claim 2, characterized in that, The traditional Chinese medicine composition consists of the following raw materials in parts by weight: 8 parts Rehmannia glutinosa, 4 parts Curculigo orchioides, 4 parts Epimedium brevicornu, 8 parts Cuscuta chinensis, 4 parts Angelica sinensis, 4 parts Cyperus rotundus, 4 parts Paeonia lactiflora, 4 parts Eucommia ulmoides, 4 parts Phellodendron chinense, and 4 parts Anemarrhena asphodeloides.

4. A method for preparing the traditional Chinese medicine composition according to any one of claims 1-3, characterized in that, Includes the following steps: Mix all the ingredients, add water and boil, filter, combine the filtrates, concentrate, dry, and pulverize to obtain the final product.

5. The preparation method according to claim 4, characterized in that, The conditions for adding water and decocting are as follows: the number of decoctions is 1-3, the amount of water added each time is 8-10 times the total mass of the medicinal materials, and the decoction time each time is 0.5-1.5 hours.

6. The preparation method according to claim 4, characterized in that, The concentration conditions are: temperature 60℃-75℃, vacuum degree -0.08 MPa to -0.10 MPa; the drying conditions are: temperature ≤70℃, vacuum degree -0.04 MPa to -0.10 MPa.

7. A pharmaceutical preparation, characterized in that, It includes the traditional Chinese medicine composition according to any one of claims 1-3 or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 4-6, as well as pharmaceutically acceptable excipients.

8. The pharmaceutical preparation according to claim 7, characterized in that, The dosage form of the pharmaceutical preparation is selected from granules, powders, decoctions, pills, tablets, capsules, mixtures, or syrups.

9. The pharmaceutical preparation according to claim 7, characterized in that, The pharmaceutically acceptable excipients are selected from one or more of the following: fillers, disintegrants, glidants, binders, flavoring agents, emulsifiers, suspending agents, sustained-release materials, propellants, dispersants, and coating materials.

10. The use of a traditional Chinese medicine composition according to any one of claims 1-3, or a traditional Chinese medicine composition prepared by any one of claims 4-6, or a pharmaceutical preparation according to any one of claims 7-9, in the preparation of a medicament for treating early-onset ovarian insufficiency.

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