Medicine for endometrial cell senescence and preparation process thereof

Abstract

The invention relates to a medicine for endometrial cell senescence and a preparation process thereof, and belongs to the technical field of traditional Chinese medicine preparations. The medicine comprises the following traditional Chinese medicine components: 10-30 parts of semen cuscutae, 5-15 parts of herba taxilli, 5-15 parts of radix dipsaci, 5-15 parts of radix angelicae sinensis, 5-15 parts of radix scutellariae, 5-15 parts of radix paeoniae alba, 5-15 parts of rhizoma chuanxiong and 1-9 parts of rhizoma atractylodis macrocephalae, and has a remarkable curative effect on endometrial cell senescence in the implantation window phase of recurrent abortion. Pharmacodynamic experiments prove that the suppository prepared from the traditional Chinese medicine disclosed by the invention can be used for remarkably improving endometrial cell senescence, especially endometrial cell senescence in an implantation window stage (WOI) of recurrent abortion (URSA). The dosage form of the medicine can be suppositories, granules, capsules and the like, pharmacodynamic experiments prove that the same technical effect can be achieved, and the preparation is simple in preparation technology and suitable for further popularization and application.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine preparations, and relates to a drug for endometrial cell senescence and its preparation process. Background Art

[0002] Recurrent spontaneous abortion (RSA) refers to the loss of pregnancy with a gestational age less than 20 weeks for two or more consecutive times, and the incidence rate is about 1% - 5%. Existing studies have shown that the causes and pathological mechanisms of RSA are extremely complex. In addition to known factors such as immune abnormalities, angiogenesis disorders, endocrine disorders, and chromosomal abnormalities, there are still about 50% of RSA patients whose causes are not clear, and they are defined by scholars as unexplained recurrent spontaneous abortion (URSA).

[0003] Cuscuta chinensis is a herbal medicine included in "Chinese Herbal Medicine". Its medicinal part is the seeds of plants such as Cuscuta chinensis, Cuscuta australis, and Cuscuta japonica, which are dicotyledonous plants of the Convolvulaceae family. Cuscuta chinensis contains glycosides, β-carotene, γ-carotene, vitamin A substances, etc. Cuscuta chinensis has a pungent, sweet, and flat taste, and can tonify the liver and kidneys, benefit the essence and marrow, nourish the muscles and strengthen the yin, strengthen the bones and tendons, boost qi and strength, and make people healthy and strong. It has good curative effects on various diseases such as early whitening of hair, soreness and weakness of the waist and knees, and loose teeth caused by deficiency of the lower energizer, and is a widely used drug in beauty prescriptions. "Compendium of Materia Medica Justice" states: "Cuscuta is a top-quality product for nourishing yin and dredging collaterals. Sweating can remove freckles on the face, and it also has the function of softening and moistening the skin."

[0004] Hyperin is a compound naturally present in many plants such as bitter almonds, bitter apricot kernels, and bitter apricots. It belongs to the class of cyanogenic glycosides and is composed of glucose, phenylalanine, and cyanide. This compound has been found in the seeds, kernels, and fruits of some plants, especially in higher amounts in almond plants. Summary of the Invention

[0005] 1. In view of the defects of the prior art, the main object of the present invention is to provide a traditional Chinese medicine composition mainly comprising 10 - 30 parts of Cuscuta chinensis, 5 - 15 parts of Taxillus chinensis, 5 - 15 parts of Dipsacus asperoides, 5 - 15 parts of Angelica sinensis, 5 - 15 parts of Scutellaria baicalensis, 5 - 15 parts of Paeonia lactiflora, 5 - 15 parts of Ligusticum chuanxiong, and 1 - 9 parts of Atractylodes macrocephala. It solves the problem of endometrial cell senescence during the implantation window of recurrent spontaneous abortion. The dosage form of the drug of the present invention can be suppository, granule, capsule, etc. Pharmacodynamic experiments have confirmed that the same technical effects can be achieved.

[0006] Specifically, the present invention adopts the following technical solutions to achieve the above object:

[0007] A drug for endometrial cell senescence, the drug is composed of the following raw materials in parts by weight: 10-30 parts of Cuscuta chinensis, 5-15 parts of Taxillus chinensis, 5-15 parts of Dipsacus asperoides, 5-15 parts of Angelica sinensis, 5-15 parts of Scutellaria baicalensis, 5-15 parts of Paeonia lactiflora, 5-15 parts of Ligusticum chuanxiong, 1-9 parts of Atractylodes macrocephala.

[0008] In a preferred embodiment of the present invention, the drug is composed of the following raw materials in parts by weight: 20 parts of Cuscuta chinensis, 10 parts of Taxillus chinensis, 10 parts of Dipsacus asperoides, 10 parts of Angelica sinensis, 10 parts of Scutellaria baicalensis, 10 parts of Paeonia lactiflora, 10 parts of Ligusticum chuanxiong, 5 parts of Atractylodes macrocephala.

[0009] The second object of the present invention provides a pharmaceutical preparation, the pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine and pharmaceutically acceptable excipients.

[0010] In a preferred implementation of the invention, the preparation is selected from granules. Further, the granules

[0011] The preparation of the agent includes the following steps:

[0012] a. Preparation of fine powder of traditional Chinese medicine

[0013] 1) Grind Cuscuta chinensis into fine powder under vacuum, and extract the volatile oil of Cuscuta chinensis; reserve for later use

[0014] 2) Crush Dipsacus asperoides, Atractylodes macrocephala, Ligusticum chuanxiong and Angelica sinensis, reflux and extract twice with ethanol above 65%, combine the filtrates, and reserve for later use;

[0016] 3) Decoct Taxillus chinensis, Scutellaria baicalensis and Paeonia lactiflora with water twice. The first decoction is for 40-60 min, and the filtrate and residue are reserved for later use; the second decoction is for 20-40 min, and combine the two decoction liquids;

[0017] 4) Combine the volatile oil in step 1), the filtrate in step 2) and the decoction liquid obtained in step 3), filter, concentrate to a clear paste with a relative density of 1.03-1.07, add ethanol to make the alcohol content above 70%, stand for 10-20 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.10-1.20 to obtain a thick paste. The vacuum degree is -0.08 MPa to -0.05 Mpa, the drying temperature is 80-90 °C, and it is pulverized through an 80-mesh sieve under vacuum

[0018] Obtain the fine powder of traditional Chinese medicine, pack it into bags, and obtain the granules;

[0019] For the above step (4), the preparation process of the capsule: Combine the fine powder of traditional Chinese medicine with pharmaceutically acceptable excipients, fill into capsules, and obtain the capsules.

[0020] For the above step (4), the preparation process of the tablets: Combine the fine powder of traditional Chinese medicine with pharmaceutically acceptable excipients, and press tablets to obtain.

[0021] For the above step (4), the pill preparation process: combine the fine powder of traditional Chinese medicine with pharmaceutically acceptable excipients, granulate to obtain pills.

[0022] For the above step (4), the decoction preparation process:

[0023] 1) Grind dodder seeds into fine powder under vacuum, extract the volatile oil of dodder seeds; reserve for later use.

[0024] 2) Crush dipsacus roots, atractylodes macrocephala, chuanxiong, and angelica sinensis, reflux and extract twice with ethanol above 65%, combine the filtrates, reserve for later use;

[0026] 3) Decoct taxillus sutchuenensis, scutellaria baicalensis, and white peony root with water twice. For the first decoction, boil for 40 - 60 minutes, reserve the filtrate and residue; for the second decoction, boil for 20 - 40 minutes, combine the two decoction liquids;

[0027] 4) Combine the volatile oil from step 1), the filtrate from step 2), and the decoction liquid obtained in step 3), filter to obtain the product.

[0028] The oral liquid preparation process: concentrate the filtrate of step (4), add appropriate excipients to obtain the oral liquid.

[0029] Any one of decoction, oral liquid, pill, or suppository.

[0030] A suppository for preventing and treating endometrial cell senescence, calculated by weight, the suppository is composed of the following raw materials in parts by weight: 10 - 30 parts of dodder seeds, 5 - 15 parts of taxillus sutchuenensis, 5 - 15 parts of dipsacus roots, 5 - 15 parts of angelica sinensis, 5 - 15 parts of scutellaria baicalensis, 5 - 15 parts of white peony root, 5 - 15 parts of chuanxiong, 1 - 9 parts of atractylodes macrocephala, 20 - 40 parts of chitosan, 10 - 30 parts of diethyl phthalate, 10 - 30 parts of colloidal silica, 30 - 50 parts of citric acid, 20 - 40 parts of sodium bicarbonate, 6 - 23 parts of butylated hydroxyanisole, and 130 - 270 parts of suppository base.

[0031] Further, in a preferred embodiment of the present invention, the dodder seeds are 20 parts, the taxillus sutchuenensis is 10 parts, the dipsacus roots are 10 parts, the angelica sinensis is 10 parts, the scutellaria baicalensis is 10 parts, the white peony root is 10 parts, the chuanxiong is 10 parts, the atractylodes macrocephala is 5 parts, the chitosan is 30 parts, the diethyl phthalate is 20 parts, the colloidal silica is 20 parts, the citric acid is 40 parts, the sodium bicarbonate is 30 parts, the butylated hydroxyanisole is 13 parts, and the suppository base is 200 parts.

[0032] Further, the suppository base contains 50 - 100 parts of carrageenan, 50 - 100 parts of glycerol, 20 - 40 parts of polyethylene glycol 4000, 10 - 30 parts of polyvinyl alcohol; preferably, the suppository base contains 75 parts of carrageenan, 75 parts of glycerol, 30 parts of polyethylene glycol 4000, and 20 parts of polyvinyl alcohol.

[0033] Further, in a preferred embodiment of the present invention, the carrageenan is selected from one of κ-type, ι-type, λ-type, γ-type, ν-type, ξ-type, and μ-type. Preferably, the carrageenan is selected from κ-type or ι-type.

[0034] Furthermore, the preparation method of the suppository is as follows:

[0035] a. Preparation of fine traditional Chinese medicine powder

[0036] 1) Cuscuta chinensis is ground into fine powder under vacuum, and the volatile oil of Cuscuta chinensis is extracted; reserved for later use

[0037] 2) Dipsacus asperoides, Atractylodes macrocephala, Ligusticum chuanxiong, and Angelica sinensis are pulverized, refluxed and extracted twice with ethanol above 65%, and the filtrates are combined and reserved for later use;

[0039] 3) Taxillus chinensis, Scutellaria baicalensis, and Paeonia lactiflora are decocted twice with water. The first decoction is for 40 - 60 min, and the filtrate and residue are reserved for later use; the second decoction is for 20 - 40 min, and the two decoction liquids are combined;

[0040] 4) The volatile oil in step 1), the filtrate in step 2), and the decoction liquid obtained in step 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.03 - 1.07, ethanol is added to make the alcohol content above 70%, allowed to stand for 10 - 20 hours, filtered, ethanol is recovered under reduced pressure and concentrated to a relative density of 1.10 - 1.20 to obtain a thick paste, the vacuum degree is -0.08 MPa to -0.05 Mpa, the drying temperature is 80 - 90 °C, and it is pulverized through an 80 - mesh sieve under vacuum to obtain fine traditional Chinese medicine powder; reserved for later use;

[0041] b. Preparation of suppository

[0042] 1) Chitosan is taken, added to water, stirred until dissolved, then colloidal silica and fine traditional Chinese medicine powder are added, diethyl phthalate is added, stirred, emulsified and crosslinked, the microspheres are centrifuged and separated, and dried to obtain drug - loaded microspheres;

[0043] 2) The drug - loaded microspheres obtained in step 2) are stirred and mixed with butylated hydroxyanisole and citric acid to obtain a mixture;

[0044] 3) Sodium bicarbonate is added to the suppository base, after mixing evenly, the mixture in step 3) is added, mixed evenly, poured into a suppository mold, cooled and solidified, and the suppository is taken out to obtain the product.

[0045] Further, in a preferred embodiment of the present invention, the preparation method of the suppository is as follows:

[0046] a. Preparation of fine traditional Chinese medicine powder

[0047] 1) Cuscuta chinensis is ground into fine powder under vacuum, and the volatile oil of Cuscuta chinensis is extracted; reserved for later use

[0048] 2) Dipsacus asperoides, Atractylodes macrocephala, Ligusticum wallichii and Angelica sinensis are pulverized, refluxed and extracted twice with 90% ethanol, and the filtrates are combined for standby.

[0049] 3) Taxillus chinensis, Scutellaria baicalensis and Paeonia lactiflora are decocted twice with water. The first decoction is for 50 min, and the filtrate and residue are reserved for standby; the second decoction is for 30 min, and the two decoction liquids are combined.

[0050] 4) The volatile oil in step 1), the filtrate in step 2) and the decoction liquid obtained in 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.05, ethanol is added to make the alcohol content 80%, allowed to stand for 15 hours, filtered, the ethanol is recovered under reduced pressure and concentrated to a relative density of 1.15 to obtain a thick paste. The vacuum degree is -0.08 MPa, the drying temperature is 85 °C, and it is pulverized through an 80-mesh sieve under vacuum to obtain fine traditional Chinese medicine powder; reserved for standby

[0051] b. Preparation of suppository

[0052] 1) Chitosan is taken, added to water, stirred until dissolved, then colloidal silica and fine traditional Chinese medicine powder are added, and diethyl phthalate is added, stirred, emulsified and crosslinked, the microspheres are centrifuged and separated, and dried to obtain drug-loaded microspheres.

[0053] 2) The drug-loaded microspheres obtained in step 2) are stirred and mixed with butylated hydroxyanisole and citric acid to obtain a mixture.

[0054] 3) Sodium bicarbonate is added to the suppository base, and after mixing evenly, the mixture in step 3) is added.

[0055] Mix evenly, pour into a suppository mold, cool and solidify, take out the suppository, and obtain it.

[0056] Furthermore, in the preferred embodiment of the present invention, the extraction method of Cuscuta chinensis volatile oil: Cuscuta chinensis is ground into fine powder under vacuum, placed in a flat-bottomed flask, wetted with an appropriate amount of distilled water, and steam-distilled for 2 - 6 h. The water-Cuscuta chinensis volatile oil distillate is collected, transferred to a separatory funnel, and extracted several times with petroleum ether at 30 - 60 °C. The extract is dried at low temperature with anhydrous sodium sulfate for 16 - 32 h and then filtered. The filtrate is rotary-evaporated to remove petroleum ether, and the volatile oil is collected for standby.

[0057] Further, in a preferred embodiment of the present invention, a vaginal suppository for preventing and treating endometrial cell senescence during the implantation window of recurrent miscarriage, the vaginal suppository is composed of the following raw materials in parts by weight: 20 parts of Cuscuta chinensis, 10 parts of Taxillus chinensis, 10 parts of Dipsacus asperoides, 10 parts of Angelica sinensis, 10 parts of Scutellaria baicalensis, 10 parts of Paeonia lactiflora, 10 parts of Ligusticum chuanxiong, 5 parts of Atractylodes macrocephala, 30 parts of chitosan, 20 parts of diethyl phthalate, 20 parts of colloidal silica, 40 parts of citric acid, 30 parts of sodium bicarbonate, 15 parts of butylated hydroxyanisole, 200 parts of suppository base; the suppository base contains 75 parts of kappa-carrageenan, 75 parts of glycerol, 30 parts of polyethylene glycol 4000, 20 parts of polyvinyl alcohol; the preparation method of the vaginal suppository is as follows:

[0058] a. Preparation of fine powder of traditional Chinese medicine

[0059] 1) Grind Cuscuta chinensis into fine powder under vacuum, place it in a 1000 ml flat-bottomed flask, add an appropriate amount of distilled water to moisten it for 4 h, perform steam distillation, collect the water-Cuscuta chinensis volatile oil distillate, transfer the distillate to a separatory funnel, extract it with petroleum ether at 45 °C in several times, add an appropriate amount of anhydrous sodium sulfate to the extract and dry it at low temperature for 24 h, then filter, rotate and evaporate the filtrate to remove petroleum ether, collect the volatile oil, and reserve it for later use;

[0060] 2) Crush Dipsacus asperoides, Atractylodes macrocephala, Ligusticum chuanxiong and Angelica sinensis, reflux and extract twice with 90% ethanol, combine the filtrates, and reserve them for later use;

[0062] 3) Decoct Taxillus chinensis, Scutellaria baicalensis and Paeonia lactiflora with water twice. Decoct for 50 min for the first time, reserve the filtrate and the residue; decoct for 30 min for the second time, and combine the two decoctions;

[0063] 4) Combine the volatile oil in step 1), the filtrate in step 2) and the decoction obtained in step 3), filter, concentrate to a clear paste with a relative density of 1.05, add ethanol to make the alcohol content 80%, let it stand for 15 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.15 to obtain a thick paste. Under a vacuum of -0.08 MPa and a drying temperature of 85 °C, crush it under vacuum and pass through an 80-mesh sieve to obtain fine powder of traditional Chinese medicine; reserve it for later use

[0064] b. Preparation of suppository

[0065] 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silica and fine powder of traditional Chinese medicine, add diethyl phthalate, stir, emulsify and crosslink, centrifuge to separate microspheres, and dry to obtain drug-loaded microspheres;

[0066] 2) Stir and mix the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture;

[0067] 3) Add sodium bicarbonate to the suppository base, mix evenly, and then add the mixture in step 3),

[0068] Mix evenly, pour into a suppository mold, cool and solidify, take out the suppositories, and you will get the product.

[0069] Furthermore, the senescence of endometrial cells in the present invention refers to the senescence of endometrial cells in the implantation window period of recurrent miscarriage.

[0070] The present invention has the following beneficial effects:

[0071] 1) The present invention provides a traditional Chinese medicine composition containing extracts of 10-30 parts of Cuscuta chinensis, 5-15 parts of Taxillus chinensis, 5-15 parts of Dipsacus asperoides, 5-15 parts of Angelica sinensis, 5-15 parts of Scutellaria baicalensis, 5-15 parts of Paeonia lactiflora, 5-15 parts of Ligusticum chuanxiong, and 1-9 parts of Atractylodes macrocephala, which has a significant curative effect on the senescence of endometrial cells in the implantation window period of recurrent miscarriage. Pharmacodynamic experiments confirm that the pharmaceutical dosage form of the present invention can be suppositories, granules, capsules and other dosage forms, and pharmacodynamic experiments confirm that the same technical effects can be achieved.

[0072] 2) Further, the suppository provided by the present invention can have a disintegration time limit within 30 minutes, which meets the requirements of the pharmacopoeia, and can stably release active ingredients, especially hyperoside, within 12 hours, and there is no sudden release phenomenon, and can be stably released in the vagina, and the treatment effect is better. Description of the Drawings

[0073] Figure 1 : The disintegration time limit of the related embodiments of the present invention. It can be seen that Examples 3-5 meet the requirements during the accelerated time.

[0074] Figure 2 : The content of hyperoside in the related embodiments of the present invention. It can be seen that under the premise of the same dosage, the comparative examples have varying degrees of degradation during storage.

[0075] Figure 3 : The release rate of hyperoside in the related embodiments of the present invention: Hyperoside in the vaginal suppositories obtained in Examples 1, 2, and 3 can be stably released within 12 hours.

[0076] Figure 4 : The pharmacodynamic experiment shows the fluorescence intensity levels of CCND2, LPARI, RBBP8, and GSN in the blank group, model group, comparative example 1 group, comparative example 2 group, and example 1 group. Specific Embodiments

[0077] The following further clarifies the present invention in combination with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. After reading the present invention, various equivalent modifications made by those skilled in the art fall within the scope protected by the claims of this application.

[0078] First, Cuscuta chinensis is the monarch drug of the suppository of the present invention, and its extraction method is explored.

[0079] Water extraction, Soxhlet extraction method and ordinary extraction method are carried out on its volatile oil, and the specific methods are as follows:

[0080] Method 1: 40 g of Cuscuta chinensis is ground into fine powder under vacuum and placed in a 1000 ml flat-bottomed flask. An appropriate amount of distilled water is added to moisten it, and steam distillation is carried out for 4 h. The water distillate of Cuscuta chinensis volatile oil is collected. The distillate is transferred to a separating funnel and extracted with petroleum ether at 45 °C in several times. The extract is added with anhydrous sodium sulfate and dried at low temperature for 24 h, then filtered. The filtrate is rotary evaporated to remove petroleum ether, and the volatile oil is collected for standby.

[0081] Method 2: Extract the volatile oil of Cuscuta chinensis by Soxhlet extraction method

[0082] 40 g of Cuscuta chinensis is ground into fine powder under vacuum, wrapped in a filter paper bag and put into a Soxhlet extractor. 200 mL of petroleum ether is added to the extraction bottle, and then it is fixed on an iron stand and placed in a constant temperature water bath at 55-60 °C. At the same time, the condensed water is refluxed into the extractor until the liquid in the extractor is colorless. The extract is taken out, added with anhydrous sodium sulfate and dried at low temperature for 24 h, then filtered. The filtrate is rotary evaporated to remove petroleum ether in the filtrate, and the volatile oil is collected and weighed.

[0083] Method 3: Extraction of Cuscuta chinensis

[0084] Take 40 g of Cuscuta chinensis powder crushed to 40 mesh and mix it evenly with 80% ethanol at a mass-to-volume ratio of 1:5-1:10. -1 , extract for 2 h, filter and collect the filtrate. Repeat the extraction 3 times, and combine the filtrates extracted 3 times. Concentrate to saturation, place at 4 °C for crystallization, collect the crystals and weigh them.

[0085] Hyperoside is used as the main index substance for detection in the above methods. The detection method refers to the Chinese Pharmacopoeia 2020 edition. At the same time, the extractant used in Method 1 is further explored, and the results are as follows:

[0086] Table 1 Contents of hyperoside extracted by different methods

[0087]

[0088] It can be seen from Table 1 that the content of hyperin obtained by Method 1 is the highest, and the use of petroleum ether as the extracting agent can extract the required flavonoid glycosides to the greatest extent. Therefore, the extraction process of the volatile oil of Cuscuta chinensis is as follows: 40 g of Cuscuta chinensis is ground into fine powder under vacuum, placed in a 1000 ml flat-bottomed flask, wetted with an appropriate amount of distilled water, and steam-distilled for 4 h. The water-distillate of the volatile oil of Cuscuta chinensis is collected, transferred to a separating funnel, and extracted with petroleum ether at 45 °C in several portions. The extract is added with anhydrous sodium sulfate and dried at low temperature for 24 h, then filtered. The filtrate is rotary-evaporated to remove petroleum ether, and the volatile oil is collected for standby. At the same time, the distillation time, drying time, and temperature of petroleum ether are controlled, and it is found that similar effects can be achieved within the ranges of Examples 1-3.

[0089] Example 1: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0090]

[0091] The preparation method of the vaginal suppository is as follows:

[0092] a. Preparation of Chinese medicine fine powder

[0093] 1) Cuscuta chinensis is ground into fine powder under vacuum, placed in a 1000 ml flat-bottomed flask, wetted with an appropriate amount of distilled water for 4 h, and steam-distilled. The water-distillate of the volatile oil of Cuscuta chinensis is collected, transferred to a separating funnel, and extracted with petroleum ether at 45 °C in several portions. The extract is added with an appropriate amount of anhydrous sodium sulfate and dried at low temperature for 24 h, then filtered. The filtrate is rotary-evaporated to remove petroleum ether, and the volatile oil is collected for standby;

[0094] 2) Dipsacus asperoides, Atractylodes macrocephala, Ligusticum wallichii, and Angelica sinensis are pulverized, reflux-extracted twice with 90% ethanol, and the filtrates are combined for standby;

[0095] 3) Taxillus chinensis, Scutellaria baicalensis, and Paeonia lactiflora are decocted twice with water. The first decoction is for 50 min, and the filtrate and residue are reserved; the second decoction is for 30 min, and the two decoctions are combined;

[0096] 4) The volatile oil obtained in step 1), the filtrate obtained in step 2), and the decoction obtained in 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.05, added with ethanol to make the alcohol content 80%, allowed to stand for 15 hours, filtered, the ethanol is recovered under reduced pressure and concentrated to a relative density of 1.15 to obtain a thick paste. The vacuum degree is -0.08 MPa, the drying temperature is 85 °C, and it is pulverized through an 80-mesh sieve under vacuum to obtain Chinese medicine fine powder; for standby

[0097] b. Suppository preparation

[0098] 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silicon dioxide and Chinese medicine powder, add diethyl phthalate, stir, emulsify and cross-link, centrifuge to separate microspheres, and dry to obtain drug-loaded microspheres;

[0099] 2) mixing the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture;

[0100] 3) adding sodium bicarbonate to the suppository base, mixing evenly, adding the mixture in step 3), mixing evenly, pouring into a suppository mold, cooling and solidifying, taking out the suppository, and obtaining the product.

[0101] Example 2: A vaginal suppository for preventing and treating endometrial cell senescence, comprising the following ingredients in parts by weight:

[0102]

[0103]

[0104] The preparation method of the vaginal suppository is:

[0105] a. Preparation of Chinese medicine powder

[0106] 1) Grind the dodder seeds into fine powder under vacuum, place it in a flat-bottom flask, add an appropriate amount of distilled water to moisten it, steam distill for 2 hours, collect the water-dodder seed volatile oil distillate, transfer the distillate to a separatory funnel, extract it with 30°C petroleum ether in batches, add anhydrous sodium sulfate to the extract and dry it at low temperature for 16 hours, then filter it, rotary evaporate the filtrate to remove the petroleum ether, collect the volatile oil and set aside.

[0107] 2) Grind Radix Dipsaci, Rhizoma Atractylodis Macrocephalae, Rhizoma Chuanxiong and Radix Angelicae Sinensis, add 90% ethanol and reflux extract twice, combine the filtrates and set aside;

[0108] 3) Add water to decoct the herb, Radix Scutellariae and Radix Paeoniae Alba twice. The first decoction is 40 minutes, and the filtrate and residue are reserved. The second decoction is 20 minutes, and the two decoctions are combined.

[0109] 4) combining the volatile oil from step 1), the filtrate from step 2) and the decoction obtained in step 3), filtering, concentrating to a clear paste with a relative density of 1.03, adding ethanol to make the alcohol content 70%, letting it stand for 10 hours, filtering, recovering ethanol under reduced pressure and concentrating to a relative density of 1.10 to obtain a thick paste, vacuum degree -0.05Mpa, drying temperature 80°C, crushing under vacuum to pass through an 80-mesh sieve to obtain a fine Chinese medicine powder; set aside;

[0110] b. Suppository preparation

[0111] 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silica and fine traditional Chinese medicine powder, add diethyl phthalate, stir, emulsify and crosslink, centrifuge to separate the microspheres, and dry to obtain the drug-loaded microspheres;

[0112] 2) Stir and mix the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture;

[0113] 3) Add sodium bicarbonate to the suppository base, mix evenly, then add the mixture in step 3),

[0114] Mix evenly, pour into a suppository mold, cool and solidify, take out the suppository to obtain.

[0115] Example 3: A vaginal suppository for preventing and treating endometrial cell senescence, calculated by weight, contains the following components:

[0116]

[0117]

[0118] The preparation method of the vaginal suppository is as follows:

[0119] a. Preparation of fine traditional Chinese medicine powder

[0120] 1) Grind dodder seeds into fine powder under vacuum, place in a flat-bottomed flask, add an appropriate amount of distilled water to moisten it, perform steam distillation for 6 h, collect the water-dodder volatile oil distillate, transfer the distillate to a separating funnel, extract it with petroleum ether at 60 °C in several times, add anhydrous sodium sulfate to the extract for low-temperature drying for 32 h and then filter, rotate and evaporate the filtrate to remove petroleum ether, collect the volatile oil for standby.

[0121] 2) Crush dipsacus root, atractylodes macrocephala, chuanxiong and angelica sinensis, reflux and extract twice with 90% ethanol, combine the filtrates for standby;

[0122] 3) Decoct taxillus chinensis, scutellaria baicalensis and white peony root twice with water, decoct for 60 min for the first time, keep the filtrate and residue for standby; decoct for 40 min for the second time, combine the two decoctions;

[0123] 4) Combine the volatile oil in step 1), the filtrate in step 2) and the decoction obtained in step 3), filter, concentrate to a clear paste with a relative density of 1.07, add ethanol to make the alcohol content 100%, stand for 20 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.20 to obtain a thick paste, vacuum degree -0.08 MPa

[0124] Drying temperature 90 °C, crush under vacuum and pass through an 80-mesh sieve to obtain fine traditional Chinese medicine powder; for standby;

[0125] b. Suppository preparation

[0126] 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silica and fine traditional Chinese medicine powder, add diethyl phthalate, stir, emulsify and crosslink, centrifuge to separate the microspheres, and dry to obtain the drug-loaded microspheres;

[0127] 2) Stir and mix the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture;

[0128] 3) Add sodium bicarbonate to the suppository base, mix evenly, then add the mixture in step 3),

[0129] mix evenly, pour into a suppository mold, cool and solidify, take out the suppository, and that's it.

[0130] Example 4: A vaginal suppository for preventing and treating senescence of endometrial cells, calculated by weight, comprising the following components:

[0131]

[0132]

[0133] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0134] Example 5: A vaginal suppository for preventing and treating senescence of endometrial cells, calculated by weight, comprising the following components:

[0135]

[0136]

[0137] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0138] Example 6: A granule for preventing and treating senescence of endometrial cells, calculated by weight, comprising the following components:

[0139]

[0140] The preparation method of the vaginal suppository is as follows:

[0141] a. Preparation of fine traditional Chinese medicine powder

[0142] 1) Grind dodder seeds into fine powder under vacuum, place in a flat-bottomed flask, add an appropriate amount of distilled water to moisten it, perform steam distillation for 6 h, collect the water-dodder seed volatile oil distillate, transfer the distillate to a separatory funnel, extract it with petroleum ether at 60 °C in several portions, add anhydrous sodium sulfate to the extract for low-temperature drying for 32 h and then filter, rotate and evaporate the filtrate to remove petroleum ether, and collect the volatile oil for standby.

[0143] 2) Crush dipsacus root, atractylodes macrocephala, chuanxiong, and angelica sinensis, reflux and extract twice with 90% ethanol, combine the filtrates, and reserve for use;

[0144] 3) Ramulus Taxilli, Radix Scutellariae, and Radix Paeoniae Alba are decocted twice with water. The first decoction lasts for 60 minutes, and the filtrate and residue are reserved; the second decoction lasts for 40 minutes, and the two decoction liquids are combined.

[0145] 4) The volatile oil obtained in step 1), the filtrate obtained in step 2), and the decoction liquid obtained in 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.07, ethanol is added to make the alcohol content 100%, allowed to stand for 20 hours, filtered, the ethanol is recovered under reduced pressure and concentrated to a relative density of 1.20 to obtain a thick paste, and the vacuum degree is -0.08 MPa.

[0146] The drying temperature is 90 °C, and it is pulverized through an 80-mesh sieve under vacuum to obtain fine traditional Chinese medicine powder.

[0147] b. Preparation of granules

[0148] Take the fine traditional Chinese medicine powder obtained in step 4), and then add dextrin or sucrose 5 times the total weight of the fine traditional Chinese medicine powder, and stir evenly to obtain.

[0149] Example 7: A vaginal suppository for preventing and treating endometrial cell senescence, calculated by weight, contains the following components:

[0150]

[0151] The preparation method of the vaginal suppository is as follows:

[0152] a. Preparation of fine traditional Chinese medicine powder

[0153] 1) Cuscuta chinensis is ground into fine powder under vacuum, placed in a flat-bottomed flask, wetted with an appropriate amount of distilled water, and steam-distilled for 6 hours. The water-Cuscuta chinensis volatile oil distillate is collected, transferred to a separatory funnel, and extracted with petroleum ether at 60 °C in several times. The extract is added with anhydrous sodium sulfate and dried at low temperature for 32 hours and then filtered. The filtrate is rotary-evaporated to remove petroleum ether, and the volatile oil is collected for standby.

[0154] 2) Dipsacus asperoides, Atractylodes macrocephala, Ligusticum wallichii, and Angelica sinensis are pulverized, reflux-extracted twice with 90% ethanol, and the filtrates are combined for standby;

[0155] 3) Ramulus Taxilli, Radix Scutellariae, and Radix Paeoniae Alba are decocted twice with water. The first decoction lasts for 60 minutes, and the filtrate and residue are reserved; the second decoction lasts for 40 minutes, and the two decoction liquids are combined.

[0156] 4) Combine the volatile oil from step 1), the filtrate from step 2), and the decoction obtained in 3), filter, concentrate to a clear extract with a relative density of 1.07, add ethanol to make the alcohol content 100%, stand for 20 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.20 to obtain a thick extract, dry at a vacuum degree of -0.08 MPa and a drying temperature of 90 °C, and pulverize through an 80-mesh sieve under vacuum to obtain fine traditional Chinese medicine powder; reserve for use;

[0157] b. Preparation of capsules

[0158] Take the fine traditional Chinese medicine powder from step 4), add 1 time of dextrin or sucrose based on the total weight of the fine traditional Chinese medicine powder, stir evenly, and fill into capsules to obtain the product.

[0159] Example 8: A tablet for preventing and treating senescence of endometrial cells, calculated by weight, contains the following components:

[0160]

[0161] The preparation method of the vaginal suppository is as follows:

[0162] a. Preparation of fine traditional Chinese medicine powder

[0163] 1) Grind dodder seeds into fine powder under vacuum, place them in a flat-bottomed flask, add an appropriate amount of distilled water to moisten them, perform steam distillation for 6 h, collect the water-dodder seed volatile oil distillate, transfer the distillate to a separatory funnel, extract it with petroleum ether at 60 °C in several portions, add anhydrous sodium sulfate to the extract for low-temperature drying for 32 h and then filter, and rotary evaporate the filtrate to remove petroleum ether, collect the volatile oil, and reserve for use.

[0164] 2) Pulverize dipsacus root, atractylodes macrocephala, chuanxiong, and angelica sinensis, reflux extract twice with 90% ethanol, combine the filtrates, and reserve for use;

[0165] 3) Decoct taxillus chinensis, scutellaria baicalensis, and white peony root twice with water, decoct for 60 min for the first time, reserve the filtrate and the residue; decoct for 40 min for the second time, and combine the two decoctions;

[0166] 4) Combine the volatile oil from step 1), the filtrate from step 2), and the decoction obtained in 3), filter, concentrate to a clear extract with a relative density of 1.07, add ethanol to make the alcohol content 100%, stand for 20 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.20

[0167] dry at a vacuum degree of -0.08 MPa and a drying temperature of 90 °C, and pulverize through an 80-mesh sieve under vacuum to obtain fine traditional Chinese medicine powder; reserve for use;

[0168] b. Preparation of tablets

[0169] Take the fine powder of traditional Chinese medicine from step 4), then add sorbitol in an amount equal to the weight of the fine powder of traditional Chinese medicine, and then add an appropriate amount of magnesium stearate for tabletting.

[0170] Comparative Example 1: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0171]

[0172]

[0173] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0174] Comparative Example 2: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0175]

[0176]

[0177] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0178] Comparative Example 3: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0179]

[0180] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0181] Comparative Example 4: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0182]

[0183] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0184] Comparative Example 5: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0185]

[0186]

[0187] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0188] Comparative Example 6: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0189]

[0190]

[0191] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0192] Comparative Example 7: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0193]

[0194]

[0195] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0196] Comparative Example 8: A vaginal suppository for preventing and treating senescence of endometrial cells, comprising the following components in parts by weight:

[0197]

[0198]

[0199] The preparation method of the vaginal suppository is the same as that described in Example 3.

[0200] I. Detection of Physical Properties

[0201] 1.1, Melting and Dissolution Time Limit

[0202] According to the melting and dissolution time limit inspection method in General Principles 0922 of the Fourth Part of Chinese Pharmacopoeia (2020 Edition). All 3 suppositories with fatty bases should be completely melted, softened or have no hard core when pressed within 30 minutes; all 3 suppositories with water-soluble bases should be completely dissolved within 60 minutes. If 1 suppository does not meet the requirements, another 3 suppositories should be taken for retest, and all should meet the requirements.

[0203] 1.2, Microbial Limit

[0204] According to the microbial limit inspection of non-sterile products in the Fourth Part of Chinese Pharmacopoeia (2020 Edition): inspected by the microbial counting method (General Principles 1105) and the control bacteria inspection method (General Principles 1106) and the microbial limit standards for non-sterile drugs (General Principles 1107), and should meet the requirements. That is, the total aerobic bacteria count of the vaginal suppository does not exceed 100 cfu / g, the total mold and yeast count does not exceed 10 cfu / g, and Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans shall not be detected.

[0205] 1.3, Hyperin Content in Cuscuta chinensis

[0206]

Content Determination

[0207] Chromatographic conditions and system suitability test: Use octadecylsilane chemically bonded silica gel as the filler; use acetonitrile - 0.1% phosphoric acid solution (17:83) as the mobile phase; the detection wavelength is 360 nm. The number of theoretical plates calculated based on the hyperoside peak should be not less than 5000.

[0208] Preparation of reference substance solution: Take an appropriate amount of hyperoside reference substance, accurately weigh it, and dissolve it in methanol to make a solution containing 48 μg per 1 ml, that is, obtain.

[0209] Preparation of test solution: Take 1 g of the fine powder of traditional Chinese medicine (sieved through No. 4 sieve) from the relevant examples, accurately weigh it, place it in a 50 ml volumetric flask, add 40 ml of 80% methanol, ultrasonically treat (power 500 W, frequency 40 kHz) for 1 hour, let it cool, add 80% methanol to the scale, shake well, filter, and take the subsequent filtrate, that is, obtain.

[0210] Determination method: Accurately pipette 10 μl each of the reference substance solution and the test solution, inject them into the liquid chromatograph, and determine, that is, obtain.

[0211] This product, calculated on the dried basis, contains hyperoside (C 21 H 20 O 12 ) not less than 0.10%.

[0212] 1.4. Accelerated test conditions

[0213] Place it in a thermostatic and humidity - controlled chamber at high temperature (40 °C) and high humidity (relative humidity 75 ± 5%) for an accelerated 6 - month test.

[0214] 1.5. Result analysis

[0215] 1.5.1. Disintegration time limit

[0216] Figure 1 The displayed disintegration time limit of the relevant implementation is shown. It can be seen that Examples 3 - 5 meet the requirements during the acceleration time and dissolve within 30 min. However, Comparative Example 5 and Comparative Example 8 cannot complete the dissolution within the specified time, which may be caused by the use of different aqueous matrix excipients and different amounts of matrix excipients.

[0217] 1.5.2. Microbiological limit

[0218] According to the fourth part of the Chinese Pharmacopoeia (2020 edition), for the microbiological limit inspection of non - sterile products, specifically as shown in Table 1:

[0219] Table 2. Microbiological limits of examples and comparative examples

[0220]

[0221]

[0222]

[0223] As can be seen from Table 2, the microbial tests of Examples 1-5 meet the requirements of the pharmacopoeia. Although Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans were not detected in Comparative Examples 3-8, during the accelerated experiment, the aerobic bacteria, molds, and yeasts in Comparative Examples 5, 6, and 8 exceeded the values specified in the pharmacopoeia and did not meet the standards for suppositories.

[0224] 1.5.3 Hyperin content and release rate in Cuscuta chinensis

[0225] Figure 2 It represents the hyperin content of relevant examples. It can be seen that on the premise of the same dosage, the hyperin in the comparative examples degraded to varying degrees during storage. The reason may be that different types of preservatives were selected, which affected the hyperin, or the dosage ratio changed irregularly, resulting in changes in the content.

[0226] Figure 3 It represents the hyperin release rate of relevant examples: the hyperin in the vaginal suppositories obtained in Examples 1, 2, and 3 can be released smoothly within 12 h, and there is basically no burst release phenomenon; Comparative Example 1 is a common suppository, and the hyperin release is only about 80%; Comparative Example 2 is a common suppository, and the hyperin is released completely within 4 h; Comparative Example 3 is a suppository, which can be continuously released within 12 h, but there is a burst release phenomenon of the drug at 6 h.

[0227] II. Pharmacodynamic experiments of the suppositories of the present invention

[0228] 2.1 Animals

[0229] Thirty SPF-grade healthy female CBA / J mice, nine male DBA / 2 mice, and three Balb / c mice, 8 weeks old, with a body weight of (20.2 ± 2.3) g, were all purchased from Zhejiang Chinese Medical University. The experimental animal certificate number is SYXK(Zhe)2021-0012. All mice were raised in the Experimental Animal Center of Zhejiang Chinese Medical University, with an environmental temperature of (24 ± 2)°C, a relative humidity of 45% ± 5%, and a light cycle of 12 h light / 12 h dark.

[0230] 2.2 Drugs and reagents

[0231] The suppositories prepared by the present invention, specifically the suppositories prepared in Example 3, Comparative Example 1, and Comparative Example 2. The model group was given blank matrix suppositories and the blank control group.

[0232] 2.3 Animal model establishment and grouping

[0233] CBA / J female mice were respectively caged and mated with Balb / c male mice and DBA / 2 male mice at a ratio of 2:1 to establish a normal pregnancy mouse model and a URSA mouse model (Reference: ZHU D, ZOU H, LIU J, et al. Inhibition of HMGB1 ameliorates the maternal-fetal interface destruction in unexplained recurrent spontaneous abortion by suppressing pyroptosis activation[J]. Front Immunol, 2021, 12: 782792). Every morning, the vagina of female mice was examined. If a vaginal plug was seen, the modeling was successful and counted as the first day of pregnancy. URSA mice were randomly divided into a model group, a blank group, an Example 3 group, a Comparative Example 1 group, and a Comparative Example 2 group. Male and female mice were administered drugs vaginally. Dosage: According to the formula "mouse dose per kilogram of body weight = general human body weight dose × (mouse dose conversion coefficient / human dose conversion coefficient)", the human body weight was 70 kg, and one pill was administered daily, each pill was about 2 g, and it was used at 8 pm. Each group of mice was administered drugs anally from the first day of pregnancy, once a day. On the 16th day of drug administration, the mice were sacrificed 12 hours after drug administration. The model group and the blank group were given blank matrix suppositories.

[0234] 2.4 Detection of the expression levels of genes related to URSA cell senescence by immunofluorescence

[0235] According to the article by Zhao Xiaoxuan et al. on the molecular mechanism of endometrial cell senescence during the implantation window of unexplained recurrent spontaneous abortion and the intervention effect of the kidney-tonifying and blood-activating method, the contents of CCND2, LPAR1, RBBP8, and GSN were detected.

[0236] 2.5 Statistical analysis

[0237] In all calculations, P < 0.05 indicates that the difference is statistically significant. All data were plotted in GraphPad Prism 9.0.

[0238] 2.6 Result analysis

[0239] Figure 4 The results of the expression showed that compared with the blank group, the fluorescence intensities of CCND2, LPARI, and RBBP8 in the model group were significantly decreased (P < 0.05), and the fluorescence intensity level of GSN was significantly increased. Compared with the Comparative Example 1 group and the Comparative Example 2 group, the expression levels of CCND2, PARI, and RBBP8 in the Example 1 group were significantly increased (P < 0.05), and the expression level of GSN was significantly decreased.

[0240] III. Pharmacodynamic experiments of other dosage forms of the present invention

[0241] The animal selection and modeling methods are carried out according to "2.1, 2.3" of the present invention, which will not be elaborated here.

[0242] 3.1 Administration and administration methods

[0243] 3.1.1 For the granule of Example 6, calculated by crude drug, the administration dosage: According to the formula "mouse dose per kilogram of body weight = general human body weight dose × (mouse dose conversion coefficient / human dose conversion coefficient)", the human body weight is 70 kg, 1 dose per day, taken in 2 times. Each group of mice was given intragastric administration starting from the 1st day of pregnancy, 2 times a day, and the administration dosage was 12.35 g / kg. On the 6th day of administration, the mice were sacrificed 12 h after administration.

[0244] Model group: Intragastrically administered the same dose of 0.9% normal saline.

[0245] Blank group: Intragastrically administered the same dose of 0.9% normal saline.

[0246] Positive control group: Aspirin 0.11 mL / 10 g.

[0247] 3.2 Detecting the expression levels of genes related to URSA cell senescence by immunofluorescence method

[0248] Detect the contents of CCND2, LPAR1, RBBP8, and GSN according to the article of Zhao Xiaoxuan et al. on the molecular mechanism of endometrial cell senescence during the implantation window of unexplained recurrent spontaneous abortion and the intervention effect of the method of tonifying the kidney and activating blood circulation. The specific operation is as follows:

[0249] The mice were perfused with 0.9% sodium chloride aqueous solution and 4% paraformaldehyde respectively until the limbs twitched and the tail curled up. Take the endometrial tissues of each group of mice and place them in 4% paraformaldehyde, and then dehydrate the tissues with 20% and 30% sucrose respectively. Prepare frozen sections with a thickness of 14 μm. Then, block the sections with an immunofluorescence blocking solution (5 g of bovine serum albumin, 0.5 mL of 30% Triton X-100, 0.05 g of sodium azide, 100 mL of 1×PBS) for 2 h, and drop the primary antibodies prepared with an antibody dilution solution (1 g of bovine serum albumin, 1 mL of 30% Triton X-100, 0.08 g of sodium azide, 100 mL of 1×PBS): rabbit anti-CCND2, GSN, LPAR1, and RBBP8 (all diluted at 1:200). After incubating overnight at 4°C in the dark, incubate with the fluorescent secondary antibody: Goat antirabbit IgG H&L (Alexa 488) (diluted at 1:200) for 2 h.

[0250] Wash the tissue 3 times with 1×PBS for 5 minutes each time, cover it with an appropriate amount of anti-quenching mounting medium containing DAPI, and seal the coverslip. Analyze using a fluorescence microscope.

[0251] 3.3 Statistical analysis

[0252] In all calculations, P < 0.05 indicates statistical significance. All data were plotted in GraphPad Prism 9.0.

[0253] 3.4 Result analysis

[0254] Figure 4 The results of the expression showed that compared with the blank group, the fluorescence intensities of CCND2, LPARI, and RBBP8 in the model group were significantly decreased (P < 0.05), and the fluorescence intensity level of GSN was significantly increased. Compared with Comparative Example 1 group and Comparative Example 2 group, the expression levels of CCND2, PARI, and RBBP8 in Example 1 group were significantly increased (P < 0.05), and the expression level of GSN was significantly decreased.

Claims

1. A drug for endometrial cell senescence, the drug is composed of the following raw materials in parts by weight: Cuscuta chinensis 10 - 30 parts, Taxillus sutchuenensis 5 - 15 parts, Dipsacus asperoides 5 - 15 parts, Angelica sinensis 5 - 15 parts, Scutellaria baicalensis 5 - 15 parts, Paeonia lactiflora 5 - 15 parts, Ligusticum chuanxiong 5 - 15 parts, Atractylodes macrocephala 1 - 9 parts.

2. The drug according to claim 1, characterized in that, The drug consists of the following raw materials in parts by weight: Cuscuta chinensis 20 parts, Taxillus sutchuenensis 10 parts, Dipsacus asperoides 10 parts, Angelica sinensis 10 parts, Scutellaria baicalensis 10 parts, Paeonia lactiflora 10 parts, Ligusticum chuanxiong 10 parts, Atractylodes macrocephala 5 parts.

3. A pharmaceutical preparation, characterized in that, The pharmaceutical preparation contains the drug described in any one of claims 1 - 2 and pharmaceutically acceptable excipients, and the preparation is selected from any one of granules, capsules, tablets, decoctions, oral liquids, pills or suppositories.

4. The preparation according to claim 3, characterized in that, In parts by weight, the suppository consists of the following raw materials in parts by weight: Cuscuta chinensis 10 - 30 parts, Taxillus sutchuenensis 5 - 15 parts, Dipsacus asperoides 5 - 15 parts, Angelica sinensis 5 - 15 parts, Scutellaria baicalensis 5 - 15 parts, Paeonia lactiflora 5 - 15 parts, Ligusticum chuanxiong 5 - 15 parts, Atractylodes macrocephala 1 - 9 parts, chitosan 20 - 40 parts, diethyl phthalate 10 - 30 parts, colloidal silica 10 - 30 parts, citric acid 30 - 50 parts, sodium bicarbonate 20 - 40 parts, butylated hydroxyanisole 6 - 23 parts and suppository base 130 - 270 parts. Further, Cuscuta chinensis 20 parts, Taxillus sutchuenensis 10 parts, Dipsacus asperoides 10 parts, Angelica sinensis 10 parts, Scutellaria baicalensis 10 parts, Paeonia lactiflora 10 parts, Ligusticum chuanxiong 10 parts, Atractylodes macrocephala 5 parts, chitosan 30 parts, diethyl phthalate 20 parts, colloidal silica 20 parts, citric acid 40 parts, sodium bicarbonate 30 parts, butylated hydroxyanisole 13 parts, suppository base 200 parts. The suppository base contains 50 - 100 parts of carrageenan, 50 - 100 parts of glycerol, 20 - 40 parts of polyethylene glycol 4000, 10 - 30 parts of polyvinyl alcohol; preferably, the suppository base contains 75 parts of carrageenan, 75 parts of glycerol, 30 parts of polyethylene glycol 4000, 20 parts of polyvinyl alcohol.

5. The preparation according to claim 5, characterized in that, The carrageenan is selected from one of κ - type, ι - type, λ - type, γ - type, ν - type, ξ - type and μ - type, and preferably, the carrageenan is selected from κ - type or ι - type.

6. The drug according to claim 1, characterized in that, The preparation method of the suppository is as follows: a. Preparation of fine traditional Chinese medicine powder 1) Grind Cuscuta chinensis into fine powder under vacuum, and extract the volatile oil of Cuscuta chinensis; reserve for later use. 2) Crush Dipsacus asperoides, Atractylodes macrocephala, Ligusticum chuanxiong and Angelica sinensis, and reflux - extract twice with ethanol with a concentration of more than 65%, Combine the filtrates and reserve for later use; for later use. 3) Decoct Taxillus sutchuenensis, Scutellaria baicalensis and Paeonia lactiflora with water twice. Decoct for 40 - 60 min for the first time, reserve the filtrate and residue; decoct for 20 - 40 min for the second time, and combine the two decoctions; 4) Combine the volatile oil in step 1), the filtrate in step 2) and the decoction obtained in step 3), filter, concentrate to a clear paste with a relative density of 1.03 - 1.07, add ethanol to make the alcohol content more than 70%, stand for 10 - 20 hours, filter, recover ethanol under reduced pressure and concentrate to a relative density of 1.10 - 1.20 to obtain a thick paste. Under a vacuum of - 0.08 MPa to - 0.05 Mpa and a drying temperature of 80 - 90 °C, crush under vacuum and pass through an 80 - mesh sieve to obtain fine traditional Chinese medicine powder; reserve for later use; b. Suppository preparation 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silicon dioxide and Chinese medicine powder, add diethyl phthalate, stir, emulsify and cross-link, centrifuge to separate microspheres, and dry to obtain drug-loaded microspheres; 2) mixing the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture; 3) adding sodium bicarbonate to the suppository base, mixing evenly, adding the mixture of step 3), mixing evenly, pouring into a suppository mold, cooling and solidifying, taking out the suppository, and obtaining; Further, preferably, the preparation method of the suppository is: a. Preparation of Chinese medicine powder 1) Grind the dodder seeds into fine powder under vacuum to extract the volatile oil of the dodder seeds; set aside 2) Grind Radix Dipsaci, Rhizoma Atractylodis Macrocephalae, Rhizoma Chuanxiong and Radix Angelicae Sinensis, add 90% ethanol and reflux extract twice, combine the filtrates and set aside; 3) Add water to decoct the herb, Radix Scutellariae and Radix Paeoniae Alba twice. The first decoction is 50 minutes, and the filtrate and residue are reserved. The second decoction is 30 minutes, and the two decoctions are combined. 4) The volatile oil from step 1), the filtrate from step 2) and the decoction obtained in step 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.05, ethanol is added to make the alcohol content 80%, left to stand for 15 hours, filtered, ethanol is recovered under reduced pressure and concentrated to a relative density of 1.15 to obtain a thick paste, vacuum degree -0.08MPa, drying temperature 85°C, crushed under vacuum and sieved through an 80-mesh sieve to obtain a fine Chinese medicine powder; set aside b. Suppository preparation 1) Take chitosan, add it to water, stir until dissolved, then add colloidal silicon dioxide and Chinese medicine powder, add diethyl phthalate, stir, emulsify and cross-link, centrifuge to separate microspheres, and dry to obtain drug-loaded microspheres; 2) mixing the drug-loaded microspheres obtained in step 2) with butylated hydroxyanisole and citric acid to obtain a mixture; 3) adding sodium bicarbonate to the suppository base, mixing evenly, adding the mixture in step 3), mixing evenly, pouring into a suppository mold, cooling and solidifying, taking out the suppository, and obtaining the product.

7. The medicament according to claim 5 or 6, characterized in that, Extraction method of Cuscuta volatile oil: Grind Cuscuta volatile oil into fine powder under vacuum, place in a flat-bottom flask, add appropriate amount of distilled water to moisten it, steam distill for 2-6 hours, collect the water-Cuscuta volatile oil distillate, transfer the distillate to a separatory funnel, extract with 30-60°C petroleum ether in batches, add anhydrous sodium sulfate to the extract and dry at low temperature for 16-32 hours, then filter, rotary evaporate the filtrate to remove the petroleum ether, collect the volatile oil and set aside.

8. A suppository for preventing and treating senescence of endometrial cells during the implantation window of recurrent spontaneous abortion, characterized in that, The suppository is composed of the following raw materials in parts by weight: 20 parts of Cuscuta, 10 parts of Morus alba, 10 parts of Dipsacus asper, 10 parts of Angelica sinensis, 10 parts of Scutellaria baicalensis, 10 parts of White Peony, 10 parts of Ligusticum chuanxiong, 5 parts of Atractylodes macrocephala, 30 parts of chitosan, 20 parts of diethyl phthalate, 20 parts of colloidal silicon dioxide, 40 parts of citric acid, 30 parts of sodium bicarbonate, 15 parts of butylated hydroxyanisole, and 200 parts of suppository base; the suppository base contains 75 parts of kappa-type carrageenan, 75 parts of glycerin, 30 parts of polyethylene glycol 4000, and 20 parts of polyvinyl alcohol; the preparation method of the suppository is as follows: a. Preparation of Chinese medicine powder 1) Cuscuta chinensis is ground into fine powder under vacuum and placed in a 1000 ml flat-bottomed flask. An appropriate amount of distilled water is added to moisten it for 4 h, and then steam distillation is carried out. The water-Cuscuta chinensis volatile oil distillate is collected. The distillate is transferred to a separating funnel and extracted with petroleum ether at 45°C in several portions. The extract is added with an appropriate amount of anhydrous sodium sulfate and dried at low temperature for 24 h, and then filtered. The filtrate is rotary evaporated to remove petroleum ether, and the volatile oil is collected. for standby; 2) Dipsacus asperoides, Atractylodes macrocephala, Ligusticum chuanxiong and Angelica sinensis are pulverized, refluxed and extracted twice with 90% ethanol, and the filtrates are combined. for standby; 3) Taxillus chinensis, Scutellaria baicalensis and Paeonia lactiflora are decocted twice with water. The first decoction is for 50 min, and the filtrate and residue are reserved; the second decoction is for 30 min, and the two decoctions are combined. 4) The volatile oil in step 1), the filtrate in step 2) and the decoction obtained in 3) are combined, filtered, concentrated to a clear paste with a relative density of 1.05, added with ethanol to make the alcohol content 80%, allowed to stand for 15 hours, filtered, the ethanol is recovered under reduced pressure and concentrated to a relative density of 1.15 to obtain a thick paste. The vacuum degree is -0.08 MPa, the drying temperature is 85°C, and it is pulverized through an 80-mesh sieve under vacuum to obtain fine traditional Chinese medicine powder; for standby b. Preparation of suppository 1) Chitosan is taken, added to water, stirred until dissolved, then colloidal silica and fine traditional Chinese medicine powder are added, and diethyl phthalate is added, stirred, emulsified and crosslinked, and the microspheres are centrifuged and separated and dried. Namely, the drug-loaded microspheres are obtained; 2) The drug-loaded microspheres obtained in step 2) are stirred and mixed with butylated hydroxyanisole and citric acid to obtain a mixture; 3) Sodium bicarbonate is added to the suppository base, and after mixing evenly, the mixture in step 3) is added, mixed evenly, poured into a suppository mold, cooled and solidified, and the suppository is taken out to obtain the product.

9. Use of the drug according to any one of claims 1-4 in the preparation of a drug for preventing senescence of endometrial cells.

10. The drug according to any one of claims 1-4, characterized in that The senescence of the endometrial cells is the senescence of endometrial cells in the implantation window period of recurrent spontaneous abortion.

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