Application of gaifeng essence in preparation of medicine for treating premature ovarian insufficiency

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of kaifeng essence to preparation of a medicine for treating premature ovarian insufficiency. The kaifeng essence has high total effective rate for adjuvant therapy of premature ovarian insufficiency and obvious clinical curative effect, and is a potential effective therapeutic drug.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and particularly relates to the application of Chufengjing in preparing a medicine for treating premature ovarian insufficiency. Background Art

[0002] The Phoenix Essence has the effects of warming and nourishing kidney yang, replenishing qi and nourishing blood. It is used for back pain, limb weakness, dizziness, tinnitus, neurasthenia, insomnia, palpitations, memory loss, loss of appetite and irregular menstruation in women caused by insufficient kidney yang and deficiency of qi and blood. The applicant's previous research has confirmed that the Phoenix Essence has significant efficacy in treating thin endometrium, increasing the thickness of the patient's endometrium, improving the blood supply of the uterine artery, and improving kidney yang deficiency.

[0003] Premature ovarian insufficiency (POI) refers to the ovarian dysfunction in women before the age of 40, manifested by oligomenorrhea or even amenorrhea, decreased estrogen levels, increased gonadotropin levels, and perimenopausal-related symptoms such as soreness of the waist and knees, dizziness and tinnitus, frequent urination at night, hot flashes and night sweats, breast tenderness, insomnia, depression or irritability, etc.; it is mainly characterized by menstrual disorders (oligomenorrhea or amenorrhea), high gonadotropin and low estrogen, and may be complicated by osteoporosis and cardiovascular and cerebrovascular diseases, seriously affecting the physical and mental health of female patients.

[0004] The global incidence of POI is 1% to 3.7%, affecting more than 14 million women of childbearing age, and the incidence is increasing year by year. There are many causes of POI, which are highly heterogeneous, and about 50% of patients have no clear cause. The known potential pathogenic factors mainly include congenital ovarian developmental abnormalities, genetic defects, autoimmune diseases, iatrogenic factors, infectious factors, and exposure to environmental pollutants.

[0005] The most commonly used method of Western medicine to treat this disease is hormone replacement therapy, which can effectively improve the patient's related clinical symptoms caused by hormone fluctuations. However, after stopping the drug, it is difficult to maintain long-term efficacy and it is easy to cause side effects of hormone drugs, aggravating the patient's condition and psychological pressure. At the same time, in recent years, researchers have been exploring new treatment methods, such as perfusing platelet-rich plasma into the ovaries, ovarian tissue transplantation, constructing artificial ovaries, artificial gametes and mitochondrial replacement therapy, etc., which provide new ways to treat POI. However, this type of treatment is limited by high cost, difficulty in practical application and ethical norms.

[0006] It adopts TCM syndrome differentiation and treatment, uses traditional special treatment methods, has definite efficacy, little damage, and is trusted by more and more patients clinically. This application uses Chu Feng Jing, which has the effect of warming and tonifying kidney yang, as a research drug to explore its therapeutic effect on premature ovarian insufficiency caused by kidney deficiency and liver depression. The applicant also studied the therapeutic effect of Chu Feng Jing on premature ovarian failure in the early stage. However, premature ovarian failure as a cause of disease, symptom manifestation, diagnostic criteria and treatment methods are obviously different from ovarian dysfunction diseases of premature ovarian insufficiency. In terms of the treatment of the latter, it does not have a definite reference significance, and in-depth and systematic research is still needed for specific types of diseases. Summary of the invention

[0007] The present invention provides the use of Chu Feng Jing in the preparation of a drug for treating premature ovarian insufficiency. Clinical trials have confirmed that the total effective rate of Chu Feng Jing in treating premature ovarian insufficiency is 96.6%, with significant clinical efficacy, providing a new approach and potentially effective therapeutic drug for the existing treatment of premature ovarian insufficiency.

[0008] The present invention provides the following technical solutions:

[0009] Application of Chufengjing in the preparation of medicines for the treatment of premature ovarian insufficiency.

[0010] Furthermore, the drug for premature ovarian insufficiency is a combination of Fentaton and Chufengjing.

[0011] Furthermore, the above-mentioned drug for treating premature ovarian insufficiency is used in the following manner:

[0012] Furthermore, patients with amenorrhea for more than 4 months should take 1 tablet of Fentaton and 20ml of Chufengjing oral liquid orally every day from the day of consultation, and patients with oligomenorrhea should take 1 tablet of Fentaton and 20ml of Chufengjing oral liquid orally from the first day of menstruation, and every 28 days is a treatment cycle. In the first 14 days, take 1 tablet of brick red tablet (containing 2mg of estradiol) + 10ml of Chufengjing oral liquid orally twice a day, and in the next 14 days, take 1 tablet of yellow tablet (containing 2mg of estradiol and 10mg of dydrogesterone) + 10ml of Chufengjing oral liquid orally twice a day. After one treatment cycle, continue the next cycle of treatment from the 29th day, and take it orally for three consecutive cycles.

[0013] Furthermore, the Chu Feng Jing is a pure Chinese medicine oral preparation; the dosage form is oral liquid, powder, granule, pill, tablet or capsule; Chu Feng Jing is made of the following ingredients in parts by weight: 6g of Amomum villosum, 5g of Cinnamon bark, 4g of Moutan bark, 5g of Acorus calamus, 7g of Aquilaria wood, 8g of Astragalus, 6g of Licorice, 4g of Ginseng, 3g of Deer Antler, 9g of White Peony Root, 8g of Psoralea corylifolia, 9g of Rubus idaeus, 5g of Nelumbo Stamen, 8g of Lycium barbarum, 14g of Alisma Orientalis, 18g of Rehmannia root, 11g of Cistanche deserticola, 15g of Dioscorea opposita, 6g of Angelica sinensis, 5g of Achyranthes bidentata, 8g of Poria, 5g of Epimedium, 7g of Cynomorium songaricum, 12g of Dog Whip, 5g of Eucommia bark, 75g of Sheep Kidney, 12g of Sheep Whip, and 150g of Chicken Fetus.

[0014] The above-mentioned Chu Feng Jing adopts the following preparation steps: a total of 28 medicinal materials of the above-mentioned quality, including Amomum villosum, cinnamon bark, peony bark, calamus, agarwood, astragalus, licorice, ginseng, deer antler, white peony root, psoralea corylifolia, raspberry, lotus stamen, wolfberry, oriental rhizome, rehmannia root, cistanche deserticola, yam, angelica, Achyranthes bidentata, Poria, epimedium, Cynomorium songaricum, dog penis, Eucommia ulmoides, sheep kidney, sheep penis and chicken fetus, are processed and crushed, and then 70% ethanol is added for percolation, the extract is collected, concentrated, fixed to volume with water, filled and sterilized.

[0015] The beneficial effects of the present invention include but are not limited to:

[0016] The present invention's use of Chu Feng Jing to treat premature ovarian insufficiency due to kidney deficiency and liver depression is an expansion of the new therapeutic application of Chu Feng Jing, and also provides an effective solution to the deficiencies of existing treatment drugs and methods for premature ovarian insufficiency. The present invention has been observed through clinical trials, and the total effective rate of Chu Feng Jing for adjuvant treatment of patients with premature ovarian insufficiency is 96.6%, with significant clinical efficacy, indicating that Chu Feng Jing has a significant clinical efficacy in the treatment of premature ovarian insufficiency due to kidney deficiency and liver depression, and can improve TCM symptoms such as soreness of waist and knees, dizziness and tinnitus, and also has a good effect on sex hormone levels, the number of antral follicles and menstrual quality. DETAILED DESCRIPTION

[0017] In order to clearly illustrate the technical features of this solution, the present invention is described in detail below through specific implementation methods. The scope of the present invention is not limited to the following embodiments. Professionals in the field can understand that various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention.

[0018] Unless otherwise specified, the instruments, reagents, materials, etc. involved in the following embodiments are conventional instruments, reagents, materials, etc. already available in the prior art and can be obtained through regular commercial channels.

[0019] 1. Preclinical research

[0020] 1.1 Summary of pharmacological research experiments

[0021] Seven pharmacological experiments were conducted on Chu Feng Jing, and the results showed that: 1) Chu Feng Jing can improve the coordinated movement and fatigue resistance of mice; 2) Continuous oral administration of Chu Feng Jing for 7 days can prolong the mice's tolerance to hypoxia and reduce oxygen consumption; 3) It can significantly increase the diameter of the rat's ear microvessels and increase the blood flow rate; 4) It can significantly increase the blood flow in the rat's carotid artery; 5) It can improve the immune function: improve the phagocytic function of mouse macrophages; can partially resist the immunosuppressive effect caused by cyclophosphamide (CTX); can promote the production of serum hemolysin; and has a significant antagonistic effect on the reduction of plaque-forming cells (PFC) caused by CTX; 6) It can significantly inhibit ADP-induced platelet aggregation in yang-deficiency rats; and enhance the body's immune function.

[0022] 1.2 Summary of toxicology studies

[0023] The acute toxicity and long-term toxicity of Chu Feng Jing were studied. The experimental results showed that Chu Feng Jing is safe and has no obvious toxic side effects.

[0024] (1) Acute toxicity

[0025] The dosage of Chu Feng Jing is equivalent to 132 times the clinical dosage of human (50kg). After 24 hours and 7 days of administration, the mice showed no abnormality in activity, their fur was shiny, their stools were normal, and their weight increased slightly, indicating that Chu Feng Jing is relatively safe.

[0026] (2) Long-term toxicity

[0027] Chu Feng Jing is equivalent to 3, 2, and 1 times the clinical dosage of human (50 kg) by oral administration. After 60 consecutive days of administration, the weight of rats increased, and no abnormalities were found in the white blood cell count, alanine aminotransferase, and urea nitrogen tests. Conventional pathological microscopy also did not find any drug-related pathological changes, indicating that long-term use of Chu Feng Jing has no obvious toxic effects.

[0028] 2. Clinical Trials

[0029] 2.1 Purpose of the study: To evaluate the efficacy and safety of Chu Feng Jing in the treatment of premature ovarian insufficiency due to kidney deficiency and liver depression.

[0030] 2.2 Test basis

[0031] The trial plan was formulated based on the "Drug Registration Management Measures", "Quality Management Standards for Drug Clinical Trials", "Drug Adverse Reaction Reporting and Monitoring Management Measures", "Guidelines for Clinical Research of New Chinese Medicines" edited by Zheng Xiaoyu, "Obstetrics and Gynecology" (8th edition edited by Xie Xing and Gou Wenli) and "Chinese Gynecology" (9th edition edited by Ma Baozhang and Qi Cong, 2nd edition edited by Zhang Yuzhen), "Standards for Diagnosis and Efficacy of Chinese Medicine Diseases" (State Administration of Traditional Chinese Medicine), "Guidelines for the Management of Premature Ovarian Insufficiency" (prepared by ESHRE), "Chinese Expert Consensus on Clinical Diagnosis and Treatment of Premature Ovarian Insufficiency" (edited by Chen Zijiang), and relevant pharmacology and toxicology experiments and clinical research data of Chufengjing.

[0032] 2.3 Diagnostic criteria

[0033] 2.3.1 Western medicine diagnostic criteria

[0034] The POI diagnostic criteria were formulated based on the "Chinese Expert Consensus on Clinical Diagnosis and Treatment of Premature Ovarian Insufficiency" published in September 2017 and the national higher medical college textbook "Obstetrics and Gynecology (9th Edition)" and other related contents:

[0035] (1) Age <40;

[0036] (2) infrequent menstruation or amenorrhea for at least 4 months;

[0037] (3) serum basal FSH>25U / L for at least 2 times (interval>4 weeks);

[0038] 2.3.2 TCM diagnostic criteria

[0039] The diagnostic criteria for POI kidney deficiency and liver depression syndrome were formulated with reference to the national planning textbook of TCM Gynecology (10th edition) for general higher education during the 13th Five-Year Plan period and the TCM Disease Diagnosis and Efficacy Standards compiled by the State Administration of Traditional Chinese Medicine in 2012:

[0040] Main symptoms: Menstrual disorders, mainly manifested as late menstruation or even amenorrhea, shortened menstrual period, and reduced menstrual volume.

[0041] Secondary symptoms: ① Weakness of waist and knees ② Dizziness and tinnitus ③ Frequent urination at night ④ Hot flashes and night sweats ⑤ Breast pain ⑥ Insomnia and nightmares ⑦ Depression or irritability.

[0042] Tongue and pulse: red tongue, thin white fur, stringy and thin or deep and thin pulse.

[0043] The main symptoms are necessary, and the diagnosis can be made if two or more secondary symptoms are present and combined with the tongue and pulse.

[0044] 2.4 Selection of subjects

[0045] 2.4.1 Inclusion criteria

[0046] (1) Meet the diagnostic criteria of traditional Chinese and Western medicine for POI;

[0047] (2) Meet the diagnostic criteria of TCM for kidney deficiency and liver depression syndrome;

[0048] (3) Age 21 years ≤ Age < 40 years;

[0049] (4) Those who can strictly prevent pregnancy during treatment;

[0050] (5) The patient had not received other treatments to improve ovarian function within 3 months before treatment;

[0051] (6) Signed informed consent and voluntary participation.

[0052] 2.4.2 Exclusion criteria

[0053] (1) Those who do not meet the inclusion criteria;

[0054] (2) patients with congenital reproductive organ abnormalities or hysterectomy;

[0055] (3) Those with endocrine diseases that affect ovulation, such as polycystic ovary syndrome, hyperprolactinemia, hyperandrogenemia, and hypothyroidism;

[0056] (4) Patients with serious cardiovascular, liver, kidney, thrombotic and blood system diseases and mental illnesses;

[0057] (5) Those who are known or suspected to have other hormone-dependent tumors, untreated endometrial hyperplasia, or a history of breast cancer and are not suitable for taking hormone drugs;

[0058] (6) Those who are currently participating in other drug clinical trials that have not yet been completed or taking other drugs that may affect this study.

[0059] Patients can be excluded if they meet any of the above criteria.

[0060] 2.4.3 Case exclusion criteria

[0061] Enrolled cases who meet any of the following criteria should be excluded:

[0062] (1) Misdiagnosis; (2) No evaluation record after treatment; (3) The use of certain prohibited drugs makes it impossible to evaluate the effectiveness.

[0063] 2.4.4 Subject dropout and withdrawal criteria

[0064] All subjects who have completed the informed consent form and are screened as eligible for enrollment have the right to withdraw from the trial at any time. Regardless of when or why they withdraw, as long as they do not complete the full observation of the clinical trial, they are considered dropouts.

[0065] (1) Withdrawal by the Investigator

[0066] For the enrolled subjects, if one of the following situations occurs during the study, the researcher may decide to withdraw them from the trial.

[0067] ① The trial was terminated due to serious adverse reactions during the trial;

[0068] ② The subjects may develop other diseases during the trial that affect the efficacy and assessment of adverse events.

[0069] ③The researcher believes that the subject is not suitable to continue receiving the study drug.

[0070] (2) Subject’s decision to withdraw

[0071] ① The subject withdraws the informed consent;

[0072] ② Those whose treatment effect is poor and who are unwilling to continue the trial treatment or whose symptoms worsen and who are unable to complete the trial;

[0073] ③Intolerable adverse events.

[0074] (3)Other reasons.

[0075] 2.4.5 Handling of dropout cases

[0076] (1) When a subject drops out, the researcher should try to keep in touch with the subject by means of telephone, appointment, follow-up, etc., and ask the subject's reason, record the last time the subject took the medication, and complete all the assessment items that can be completed.

[0077] (2) For subjects who withdraw or interrupt the study, the final evaluation (end-of-treatment visit) should be completed as much as possible, and the evaluation process, results, and description of the reasons for the subject's withdrawal must be recorded in detail in the original data and case report form.

[0078] 2.4.6 Test termination criteria

[0079] (1) The ethics committee, sponsor or investigator believes that the investigational drug has significant safety risks;

[0080] (2) The sponsor may terminate the study for any scientific, medical or ethical reasons, but the rights, safety and health of the subjects must be fully considered;

[0081] (3) Other reasons that the sponsor or investigator determines are inappropriate to continue the trial.

[0082] 2.5 Treatment options

[0083] 2.5.1 Specifications of investigational drugs

[0084] Medicine: Chu Feng Jing; Specification: 10 vials per box, 10 ml each.

[0085] 2.5.2 Packaging of investigational drugs

[0086] Mark “For clinical trial use only” on the smallest unit (box) of commercial packaging.

[0087] 2.5.3 Drug distribution and storage

[0088] 2.5.3.1 Distribution and recovery of investigational drugs

[0089] After the subjects are selected, the trial drug manager (responsible for the storage, distribution, recovery, record and return or recovery of trial drugs) will distribute the drugs, calculate the number of boxes distributed at each visit based on the menstrual situation, recover the remaining drugs during the follow-up, and fill in the "Drug Distribution and Recovery Record Form" in time according to the subjects' medication situation. After all the trials are completed, the drug manager will be responsible for returning the remaining drugs to the sponsor in a centralized manner and filling in the "Clinical Trial Drug Distribution and Recovery Registration Form" for filing.

[0090] 2.5.3.2 Storage of experimental drugs

[0091] A special counter shall be set up to store the trial drugs, which shall be managed uniformly by the drug administrator, who shall fill in the "Clinical Trial Drug Storage Record Form".

[0092] 2.5.4 Medication method and course of treatment

[0093] Control group: oral Fentaton; Dosage: Patients with amenorrhea for more than 4 months should take 1 tablet orally every day from the day of consultation, and patients with oligomenorrhea should take 1 tablet orally every day from the first day of menstruation. Every 28 days is a treatment cycle. For the first 14 days, take 1 tablet of brick red tablet (containing 2mg of estradiol) orally every day, and for the next 14 days, take 1 tablet of yellow tablet (containing 2mg of estradiol and 10mg of dydrogesterone) orally every day. After one treatment cycle, continue the next cycle of treatment from the 29th day, and take it orally for three consecutive cycles. The treatment group combined the treatment of Chufengjing with the treatment plan of the control group. Usage: Oral. 10ml at a time, twice a day; take it for 3 menstrual cycles.

[0094] 2.5.5 Provisions for combined medication

[0095] During the trial, hormones and Chinese patent medicines related to this disease shall not be used.

[0096] 2.5.6 Determination of Subject Compliance

[0097] During clinical trials, the subjects' compliance mainly depends on taking medications as prescribed. The subjects should fully understand the importance of taking medications on time, take medications strictly as prescribed, and avoid adding other treatments on their own.

[0098] The criteria for determining subject compliance: Compliance with trial medication = (amount of medication dispensed - amount not taken) / amount that should be taken × 100%.

[0099] Patient compliance between 80% and 120% was considered good.

[0100] 2.6 Research Process

[0101] All indicators were observed in the first menstrual cycle after enrollment and the first menstrual cycle after stopping treatment; ① TCM syndrome score (see Appendix 9 for TCM syndrome score standards); ② Serum sex hormone evaluation indicators: electrochemiluminescence method was used to draw venous blood in the fasting state 3 days before and after treatment to measure serum FSH, LH, and E2; ③ Anti-Mullerian hormone (AMH) was detected simultaneously with serum sex hormones; ④ Double ovarian antral follicle technique (AFC) was measured by ultrasound on the third day of menstruation before and after treatment; ⑤ Menstrual status score.

[0102] Visit 1: Screening & Enrollment Period (Baseline)

[0103] (1) Obtain written informed consent;

[0104] (2) Obtain demographic information, basic characteristics of the subjects, medical history, past medical history, other current diseases, and treatment status;

[0105] (3) Conduct physical examinations and collect data on vital symptoms and signs;

[0106] (4) Fill in the TCM syndrome score sheet and menstrual status score sheet;

[0107] (5) Perform blood routine, urine routine, urine pregnancy, liver function (ALT, AST, TBIL, γ-GT, ALP), renal function (BUN, Cr), and electrocardiogram examinations;

[0108] (6) Sex hormone levels (FSH, LH, E2), anti-Mullerian hormone (AMH), and ultrasound examination;

[0109] (7) Review inclusion / exclusion criteria;

[0110] (8) Distribute drugs and fill in the “Drug Distribution and Recovery Record Form”.

[0111] Visit 2: Follow-up period (one menstrual cycle of treatment)

[0112] (1) Count the unused study drugs returned by the subjects and record them in the Drug Distribution and Recovery Record Form;

[0113] (2) Collect data on vital symptoms and signs;

[0114] (3) Fill in the TCM syndrome score sheet and menstrual status score sheet;

[0115] (4) record concomitant medications and adverse events;

[0116] (5) Distribute the next cycle’s drugs and fill in the “Drug Distribution and Recovery Record Form”.

[0117] Visit 3: Follow-up period (two menstrual cycles of treatment)

[0118] (1) Count the unused study drugs returned by the subjects and record them in the Drug Distribution and Recovery Record Form;

[0119] (2) Collect data on vital symptoms and signs;

[0120] (3) Fill in the TCM syndrome score sheet and menstrual status score sheet;

[0121] (4) record concomitant medications and adverse events;

[0122] (5) Distribute the next cycle’s drugs and fill in the “Drug Distribution and Recovery Record Form”.

[0123] Visit 4: Follow-up period (three menstrual cycles of treatment)

[0124] (1) Count the unused study drugs returned by the subjects and record them in the Drug Distribution and Recovery Record Form;

[0125] (2) Conduct physical examinations and collect data on vital symptoms and signs;

[0126] (3) Fill in the TCM syndrome score sheet and menstrual status score sheet;

[0127] (4) Perform blood routine, urine routine, urine pregnancy, liver function (ALT, AST, TBIL, γ-GT, ALP), renal function (BUN, Cr), and electrocardiogram examinations;

[0128] (5) Sex hormone levels (FSH, LH, E2), anti-Mullerian hormone (AMH), and ultrasound examination;

[0129] (6) Record concomitant medications and adverse events.

[0130] The clinical trial flow chart is shown in Table 1.

[0131] Table 1 Clinical trial flow chart

[0132]

[0133]

[0134] Remark:

[0135] 1. Blood routine: white blood cells, neutrophils, lymphocytes, red blood cells, hematocrit, hemoglobin, platelets. 2. Urinalysis: urine protein, urine occult blood, urine red blood cells, urine white blood cells, urine specific gravity.

[0136] 2.7 Efficacy evaluation

[0137] 2.7.1 Observation indicators

[0138] (1) Demographic data and basic characteristics of the subjects: age, ethnicity, marital status, menstruation and fertility status, etc.

[0139] (2) General clinical data: course of disease, medical history, comorbidities and medication, and allergy history.

[0140] (3) Efficacy indicators:

[0141] ① TCM syndrome scoring: Scoring is based on the “Standards for Diagnosis and Efficacy of TCM Diseases” compiled by the State Administration of Traditional Chinese Medicine in 2012.

[0142] ② Serum sex hormone levels: including FSH, LH, and E2;

[0143] ③ Anti-Mullerian hormone (AMH);

[0144] ④Ultrasound: number of antral follicles;

[0145] ⑤ Menstrual status score;

[0146] (4) Safety observation indicators: ① Adverse events were observed at any time; ② Vital signs (blood pressure, respiration, heart rate); ③ Abnormal changes in laboratory indicators before and after treatment: blood routine, urine routine, liver function (ALT, AST, TBIL, γ-GT, ALP), renal function (BUN, Cr), electrocardiogram.

[0147] 2.7.2 Determination of efficacy and safety

[0148] 2.7.2.1 Statistics of efficacy indicators

[0149] Corresponding statistical methods were used to perform statistical analysis on TCM syndrome scores, sex hormone levels (FSH, LH, E2), anti-Mullerian hormone (AMH), number of antral follicles, and menstrual status scores.

[0150] 2.7.2.2 TCM syndrome efficacy evaluation criteria

[0151] It is formulated with reference to the relevant evaluation standards in the "Guiding Principles for Clinical Research of New Chinese Medicines" and the "Standards for Diagnosis and Efficacy of Clinical Diseases in Traditional Chinese Medicine", and combined with clinical practice.

[0152] Cured: All clinical symptoms basically disappeared after treatment, n≥95%;

[0153] Markedly effective: All clinical symptoms are significantly relieved after treatment, 70%≤n<95%;

[0154] Effective: All clinical symptoms are relieved after treatment, 30%≤n<70%;

[0155] Ineffective: clinical symptoms were not improved or worsened after treatment, n<30%.

[0156] Calculation formula (nimodipine method): therapeutic index (n) = (score before treatment - score after treatment) / score before treatment × 100%.

[0157] 2.7.2.3 Comprehensive efficacy criteria

[0158] Cure: n≥95%, the patient's menstruation returns to normal after stopping the drug, clinical symptoms are significantly improved or completely disappear, serological indicators basically return to normal, and FSH <10IU / L;

[0159] Markedly effective: 70%≤n<95%. After stopping the drug, the patient's menstruation basically returns to normal, the clinical symptoms are relieved, the serological indicators are significantly improved, and 10IU / L≤FSH<25IU / L is met;

[0160] Effective: 30%≤n<70%. After stopping the drug, the patient's menstrual condition is slightly improved, but the menstrual period is shorter or the menstrual volume is less. The clinical symptoms are slightly relieved, the serological indicators are slightly improved, and FSH is ≥25IU / L, but it is more than 30% lower than before.

[0161] Ineffective: n<30%, the patient's menstrual condition did not improve or menstruation did not occur after stopping the drug, clinical symptoms did not improve or worsened, serological indicators basically did not improve, FSH value did not change significantly, or increased compared with the previous value.

[0162] Note: Total effective rate = (number of clinically cured cases + number of markedly effective cases + number of effective cases) / total number of cases × 100%

[0163] 2.7.2.4 Safety evaluation criteria

[0164] Level 1: safe, without any adverse reactions;

[0165] Level 2: relatively safe. If there are adverse reactions, no treatment is required and the medication can be continued;

[0166] Level 3: There are safety issues and moderate adverse reactions, but the drug can continue to be administered after treatment;

[0167] Grade 4: The trial was terminated due to serious adverse reactions.

[0168] 2.8 Observation of adverse events

[0169] 2.8.1 Relevant definitions

[0170] Adverse events: The term adverse events covers the appearance or worsening of any signs, symptoms, syndromes or diseases that occur in subjects during clinical study observation and that may affect the health of the subjects. The term also includes clinically relevant conditions found during laboratory or other diagnostic procedures. Adverse events may be: new diseases; worsening of symptoms or signs of the treatment state, or worsening of concomitant diseases; effects of control drugs; unrelated to participation in the trial; or a combination of one or more factors.

[0171] Serious adverse events are the following adverse events that occur at any dose of the trial drug or at any time during the observation period, including: leading to death; immediately threatening life; requiring hospitalization or prolonging hospitalization; causing disability; causing congenital malformations; having important medical significance (referring to those events that are not immediately life-threatening or cause death or require hospitalization, but may endanger the patient or require measures to prevent one of the consequences defined above); requiring medical treatment to prevent permanent injury or damage.

[0172] Adverse drug reaction: harmful and undesirable reactions that are causally related to the use of drugs and occur during the normal use of drugs at prescribed doses. In clinical trials of a new drug or a new use of a drug, when the therapeutic dose has not yet been determined, all harmful and undesirable reactions that are causally related to the use of the drug should also be considered adverse drug reactions.

[0173] 2.8.2 Adverse events

[0174] 2.8.2.1 Recording of adverse events

[0175] During the study, the Adverse Event Record Form should be filled out truthfully to record the time of occurrence, severity, duration, effective measures taken and outcomes of adverse events.

[0176] 2.8.2.2 Severity Determination

[0177] Mild: Usually transient and does not affect normal daily activities.

[0178] Moderate: Considerable discomfort that interferes with normal daily activities.

[0179] Severe: Inability to carry out normal daily activities.

[0180] 2.8.2.3 Determination of causal relationship with drugs

[0181] The researcher should truthfully record the time, severity, duration, measures taken and outcome of adverse events that occurred during the subject's medication, and determine the relationship between the adverse event and the trial drug. The association between adverse events and trial drugs should be evaluated according to the five-level classification method of "definitely related, very likely related, possibly related, suspicious, and unlikely related". The first three levels are judged to be related to the trial drug. When calculating the adverse reaction rate, the total of the three is used as the numerator, and the total number of subjects used to evaluate safety is used as the denominator. See Table 2 for details.

[0182] Table 2 Causality criteria for adverse reactions

[0183]

[0184] 2.8.2.4 Handling of adverse events

[0185] (1) Reporting method

[0186] Any adverse events, such as subjective discomfort of patients and abnormal laboratory tests, must be taken seriously, carefully analyzed, and immediate measures must be taken to protect the safety of the subjects.

[0187] (2) Processing Procedure

[0188] Record the details in the case report form and record its persistence, outcome, disappearance, etc. as appropriate.

[0189] (3) Reporting and handling of serious adverse events

[0190] Any serious adverse event occurring during the trial must be reported immediately to the principal investigator, ethics committee and sponsor by filling out the “Serious Adverse Event Report Form” to record the time of occurrence, severity, duration, measures taken and outcome of the serious adverse event.

[0191] Treatment measures: When a patient experiences a serious adverse event, he or she should withdraw from the clinical trial. The principal investigator of the research unit should take appropriate measures based on the drug and the symptoms that occur. The researcher should record the date, treatment status, and results in detail on the case report form and sign it.

[0192] (4) Follow-up of adverse events

[0193] Adverse events that are not resolved at the end of the trial or at the time of early withdrawal of the subject must be followed up until any of the following conditions are reached:

[0194] The event is resolved; the event is stable; the event returns to baseline levels (if baseline values ​​are available); the event can be attributed to drugs other than the investigational drug or factors unrelated to the study behavior, or when more information is unlikely to be obtained (the patient or health care provider refuses to provide more information, or there is evidence that the patient is lost to follow-up despite best efforts).

[0195] 2.9 Data Management

[0196] 2.9.1 Data recording and entry

[0197] The case report form is filled out by the researcher, and each selected case must complete the case report form. The dropout time, cause of dropout and treatment should be recorded in detail for the dropout case. The completed case report form is reviewed by the clinical monitor and the main person in charge of the research unit, and then handed over to the data manager for data entry and management. Data entry and management are the responsibility of the data manager. The data manager establishes the Epidata (v3.1) database based on the case report form for data entry. The database name must be standardized, easy to read, easy to find, and ensure its correctness, security and confidentiality. All data are entered in duplicate by two data entry personnel. After the data is entered and checked in duplicate, the data manager formulates the range check and logic check content of the data according to the range and relationship of the values ​​of each indicator in the case report form, and writes the corresponding computer program to verify the data. All questions generated during the data verification are promptly notified to the monitor in the form of a question form, and the researcher is required to answer.

[0198] After the data entry and verification are completed as required, the original case report form can be archived in the order of numbering, and the search directory can be filled in for reference. Electronic data files are classified and stored with multiple backups to prevent damage.

[0199] All file transfers are specially recorded and signed accordingly and are properly preserved in accordance with GCP regulations.

[0200] 2.9.2 Data Locking

[0201] After the data is reviewed and the database is confirmed to be correct, the principal investigator, sponsor, and statistical analyst will lock the data. The locked data file will not be modified. Problems found after the data is locked will be corrected in the statistical analysis program after confirmation.

[0202] 2.9.3 Data Processing

[0203] After all the research data are entered into the database and locked, and the statistical analysis is completed, the statistical analyst will write a statistical analysis report and hand it over to the principal investigator of the trial to write a research report.

[0204] 2.9.4 Data Encoding

[0205] The DM in the data center performs medical coding, including adverse events (AE). Adverse events will be coded according to the MedDRA21.0 dictionary. During the coding process, if any medical terminology is improper, inaccurate, or vague and cannot be coded, the DM can question the researcher. Before the database is locked, medical experts will review the medical coding.

[0206] 2.9.5 Data Quality Check

[0207] After the data cleaning is completed, non-project team members within the data management department will check the data quality, conduct 100% inspection on key data (important data determined by the research plan), select a certain range of non-key data for spot checks, and form a final data quality inspection report.

[0208] 2.9.6 Data Review

[0209] After the database is cleaned, the data manager writes a "Data Audit Report" for the data verification meeting. The audit report mainly records the number of cases enrolled, dropouts, cases excluded, deviations or violations of the protocol, compliance data, combined medication, adverse events and other data related to the evaluation indicators.

[0210] At the data review meeting, the analysis group divisions were discussed and determined based on the content of the review report.

[0211] 2.10 Statistical analysis

[0212] 2.10.1 Statistical analysis plan

[0213] SPSS 25.0 statistical software was used for data analysis. Measurement data were expressed as mean ± standard deviation (x ± s), independent sample t test was used for comparison between the two groups, and paired t test was used for comparison within the group; count data were expressed as rate, and intergroup comparison was performed using χ 2 P<0.05 was considered statistically significant.

[0214] 2.10.11 Statistical analysis content

[0215] (1) Baseline and demographic information

[0216] The demographic information, general characteristics of the subjects, general clinical data, vital signs, and baseline conditions of efficacy indicators were collected.

[0217] (2) Efficacy evaluation

[0218] ① TCM syndrome scores before and after treatment;

[0219] ② Sex hormone levels (FSH, LH, E2) before and after treatment;

[0220] ③ Anti-Mullerian hormone (AMH) before and after treatment;

[0221] ④ Number of antral follicles before and after treatment (pieces);

[0222] ⑤ Menstrual status score before and after treatment.

[0223] 2.10.4 Safety evaluation

[0224] Safety evaluation was analyzed on the SS data set. According to the requirements of adverse reaction relevance, the incidence of adverse events, adverse reactions, important adverse events, serious adverse events, and serious adverse reactions was calculated, and the list detailed the name, occurrence time, severity, relationship with the trial drug, whether the patient withdrew due to adverse events, etc.

[0225] Calculate the incidence of combined medication and list in detail the drug name, usage and dosage, start time, end time, etc. of the combined medication.

[0226] Calculate the changes in laboratory test, electrocardiogram and other examination results before and after treatment; list in detail the laboratory test, electrocardiogram and other results that were normal before treatment and abnormal after treatment, and abnormal before treatment and abnormal after treatment. Calculate the number of cases and conversion rate of laboratory test indicators "normal to abnormal" before and after the test. List in detail the abnormal cases and clinical explanations of laboratory indicators, electrocardiograms, and physical examinations.

[0227] 2.11 Clinical Research

[0228] The trial adopted a single-center (the clinical research work plan was completed by Hongjitang Traditional Chinese Medicine Hospital), open (open trial, no blinding) clinical research.

[0229] Clinical trial results: A total of 61 female subjects were included, including 31 in the control group and 30 in the treatment group; there was no statistically significant difference in the general data of the two groups (P>0.05), and they were comparable. The data results of each indicator are detailed in Table 3-Table 8.

[0230] Table 3 Comparison of total TCM syndrome scores between the control group and the three treatment groups

[0231]

[0232] Table 4 Comparison of sex hormone levels (FSH, LH, E2) between the control group and the treatment group

[0233]

[0234] Table 5 Comparison of anti-Mullerian hormone (AMH) between the control group and the treatment group

[0235]

[0236] Table 6 Comparison of the number of antral follicles before and after treatment

[0237]

[0238] Table 7 Comparison of total menstrual scores before and after treatment

[0239]

[0240] Table 8 Comparison of clinical efficacy [n (%)]

[0241]

[0242] The results of the present invention show that after treatment, the total score of sexual TCM syndromes in the two groups was compared, and both the control group and the treatment group were significantly improved, and the improvement effect of the treatment group was better, as shown in Table 3; by comparing the sex hormone levels, the FSH and LH of the two groups were reduced, and the E2 level was increased, and the LH and FSH levels of the treatment group were lower than those of the control group, and the E2 level was higher than that of the control group (P<0.05), as shown in Table 4 and Table 5. There was no significant difference in AMH between the two groups before treatment (P>0.05); the number of antral follicles in the two groups was significantly increased compared with the control group (P<0.05), and the treatment group had a better effect, as shown in Table 6; the menstrual quality of the two groups was significantly improved compared with before treatment, and the effect of the treatment group was greater than that of the control group (P<0.05), as shown in Table 7. The total effective rate of the control group was 74.2%, and the total effective rate of the treatment group was 96.6%, which was significantly higher than that of the control group, and the difference was statistically significant (P<0.05), as shown in Table 8. It shows that the clinical effect of Chufengjing in treating premature ovarian insufficiency with kidney deficiency and liver depression is obvious, and it can improve TCM symptoms such as soreness of waist and knees, dizziness and tinnitus, and it also has a good effect on sex hormone levels, the number of antral follicles and menstrual quality.

[0243] There were no adverse reactions during the trial.

[0244] Appendix 9: TCM Syndrome Scoring Standards

[0245]

[0246]

[0247] The above is only the embodiment of the present application, and the protection scope of the present application is not limited by these specific embodiments, but is determined by the claims of the present application. For those skilled in the art, the present application can have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the technical ideas and principles of the present application should be included in the protection scope of the present application.

Claims

1. Application of Chufengjing in the preparation of drugs for the treatment of premature ovarian insufficiency.

2. The use according to claim 1, characterized in that: The drug for premature ovarian insufficiency is a combination of Fentaton and Chufengjing.

3. The application according to claim 1, characterized in that: The Chu Feng Jing is a pure Chinese medicine oral preparation; the dosage form is oral liquid, powder, granule, pill, tablet or capsule; The described young phoenix essence is prepared from the following ingredients in parts by weight: 6g of Amomum villosum, 5g of Cinnamon bark, 4g of Moutan bark, 5g of Acorus calamus, 7g of Aquilaria sinensis, 8g of Astragalus, 6g of Licorice, 4g of Ginseng, 3g of Deer Antler, 9g of White Peony Root, 8g of Psoralea corylifolia, 9g of Rubus idaeus, 5g of Lotus Stamen, 8g of Lycium barbarum, 14g of Alisma orientalis, 18g of Rehmannia root, 11g of Cistanche deserticola, 15g of Dioscorea opposita, 6g of Angelica sinensis, 5g of Achyranthes bidentata, 8g of Poria, 5g of Epimedium, 7g of Cynomorium songaricum, 12g of Dog Whip, 5g of Eucommia bark, 75g of Sheep Kidney, 12g of Sheep Whip, and 150g of Chicken Fetus.

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