CN101190265A - A kind of medicine for treating endometriosis and preparation method thereof - Google Patents

Abstract

The invention relates to a medicine for treating endometriosis, which is prepared from the following raw materials in parts by weight: rhubarb 2.80-3.20, turtle shell 2.80-3.20 and amber 1.30-1.70. The invention also discloses a method for producing the medicine for treating endometriosis. The drug removes blood stasis and unblocks the internal organs, and mainly treats endometriosis, with symptoms such as dysmenorrhea, pelvic pain, anal swelling, backache and constipation, dark tongue or ecchymosis, thin fur, thready or astringent pulse; Compared with similar clinical efficacy, but fewer adverse reactions, easy to be accepted by patients.

Description

A kind of medicine for treating endometriosis and preparation method thereof

technical field

The invention relates to a traditional Chinese medicine preparation, in particular to a medicine for treating endometriosis and a preparation method thereof.

Background technique

Endometriosis is a common gynecological disease, but also a refractory disease. Can cause severe dysmenorrhea, pelvic pain, infertility and menstrual disorders. The incidence of endometriosis in the population is about 6-8%. The incidence rate is 2-22% in asymptomatic women, 40-60% in women with dysmenorrhea, 20-30% in infertile women, and about 10- Endometriosis was diagnosed postoperatively in 25% of patients. Since the etiology of endometriosis has not been fully elucidated so far, although there are many treatment methods, the effect is not very satisfactory.

The uterus, also known as the woman's cell, was first recorded in the "Emperor's Internal Classic". Because the function is different from the zang-organs storing but not purging, and also different from the fu-organs purging but not storing, it has the characteristics of both storing and purging, and periodically storing and purging, so it is called the extraordinary and permanent fu-organs in "Suwen · Five Zang-Parts".

There is no record of the disease name of endometriosis in the medical records of the past dynasties of traditional Chinese medicine, but according to its pathogenesis and clinical manifestations, endometriosis belongs to the categories of "dysmenorrhea", "abdominal lumps" and "infertility" in traditional Chinese medicine, and the pathological essence is " blood stasis". "On the Origin of Various Diseases" has the following description of "blood lumps": "Blood lumps, the woman's menstrual flow is new, and she stops before the full number of days. She eats too much, and the five grains are full of energy, which overflows into other organs; Insufficient, unregulated breathing and self-work, unsteady blood flow, left and right between the stomach and intestines, lingering knots, cold and heat inside, meeting with moon water, forming a lump of blood. Lumbago, can not pitch, under the transverse bone Accumulated qi, firm as a stone, acute pain in the lower abdomen, back pain, deep to the waist and lower abdomen, if the wind is cold in the shade, the door is slamming, the moon water is intermittent, and the disease does not come at all. This disease makes people childless. .” The pathogenesis and clinical manifestations described here are similar to those of endometriosis. "Women's Encyclopedia of Good Prescriptions" pointed out: "Those with blood stasis in the abdomen of a woman, due to menstrual closure, or the remaining blood is not exhausted, or the stasis caused by wind and cold is not clear for a long time, it is a syndrome of harmony." Jingyue Quanshu · Women's Regulations · Symptoms and Masses" pointed out: "Blood stagnation is the only symptom that women have. Anger and anger injure the liver, the qi is reversed and the blood stays, or the accumulation of fatigue is weak, and the qi is weak and does not move. When the blood is always moving, the remaining blood is not clean, but once there is a reverse, it will stay and accumulate day by day and gradually become a disease.” . What is described here is quite similar to the modern pathogenesis of endometriosis.

Endometriosis is a hot topic in the field of gynecology at home and abroad. There are many different theories about the pathogenesis of endometriosis, but the dominant theory is still that endometrial cells grow back with menstrual blood. The endometrial cells that flow back into the peritoneal cavity with menstrual blood need to go through the "trilogy" of adhesion, invasion, and angiogenesis before they can finally develop and form endometriosis lesions. traits, and even genetic differences. However, the influence of hormones, various enzymes in the local environment, and various cytokines related to human immunity also play an important role. Therefore, at present, many studies focus on the aspects of immune level, angiogenesis and genetic inheritance. The progress of basic research will promote the progress of clinical treatment methods. Anti-adhesion, anti-invasion, and anti-angiogenic drugs may be gradually applied in clinical practice. Cytokine regulation, receptor intervention and gene therapy are also expected to become new treatment methods.

At present, the clinical treatment of endometriosis mostly adopts expectant therapy, surgical treatment and drug treatment. Expectant therapy is suitable for patients with mild symptoms. Surgical therapy can use traditional surgical methods and laparoscopic surgery to remove lesions, separate adhesions, restore pelvic anatomy, and relieve symptoms. Conservative surgery can preserve and improve reproductive function, but the postoperative recurrence rate is high. Radical surgery is suitable for patients with severe symptoms, ineffective medication and conservative surgery, and no fertility requirements, and is not suitable for young women. Drug treatment mostly adopts "pseudomenopausal therapy" and "drug-induced oophorectomy", and the representative drugs are danazol, gesttriene and leuprolide. However, drug treatment has a high recurrence rate and many adverse reactions, and the clinical efficacy is not satisfactory. In recent years, the use of traditional Chinese medicine to treat endometriosis has become more and more accepted by patients, and many doctors have also conducted in-depth research on this. Most of the doctors use the method of promoting blood circulation to remove blood stasis, but around the cause and pathogenesis of the disease, there are methods of soothing the liver and promoting blood circulation to remove blood stasis, clearing heat and promoting blood circulation to remove blood stasis, nourishing the kidney and promoting blood circulation to remove blood stasis, removing blood stasis and dredging the internal organs, warming the meridian, promoting blood circulation and removing blood stasis In addition, it also proposes a variety of methods and programs such as syndrome differentiation, external treatment (enema, external application of acupoints, acupuncture and massage), and combination of internal and external. Neiyi Tablet, which embodies the method of removing blood stasis and dredging the internal organs, has similar clinical curative effect on endometriosis compared with western medicine danazol, but has fewer adverse reactions and is easy to be accepted by patients.

To sum up, the traditional Chinese medicine for treating endometriosis currently on the market is still rare.

Contents of the invention

In view of the above problems, the purpose of the present invention is to provide a kind of Chinese medicine preparation and preparation method thereof that has better effect in treating endometriosis.

To achieve the above object, the present invention takes the following technical solutions:

A medicine for treating endometriosis provided by the present invention is characterized in that it is made of raw materials containing the following ratios by weight and number:

rhubarb 2.8～3.2 Turtle shell 2.8～3.2 Amber 1.3～1.7

The preferred raw material formula of the present invention is: be made up of the raw material that contains following ratio of parts by weight:

rhubarb 3 Turtle shell 3 Amber 1.5

The method for producing the medicine for treating endometriosis provided by the present invention comprises the following steps:

1) Take the raw materials of the following weight and number ratios:

rhubarb 2.8～3.2 Turtle shell 2.8～3.2 Amber 1.3～1.7

2) Soak rhubarb plus 85% ethanol in 8 times amount for 1 hour, reflux extraction three times, each time for 1 hour, filter, combine the filtrate for 3 times, recover ethanol and concentrate under reduced pressure to a relative density of 1.08-1.10 (25°C) ;

3) Turtle shells, oysters, Panax notoginseng, and Loulu add 10 times the amount of water to extract four times, each time for 8 hours, filter, combine the filtrates, and concentrate under reduced pressure to a relative density of 1.08-1.10 (25°C).

4) Combine the concentrates of steps 2) and 3), add amber, mix well, dry in vacuum below 75°C and pulverize into fine powder,

5) Add appropriate amount of auxiliary materials to the fine powder in step 4), mix well, make granules, press into 1000 tablets, coat with film, and obtain the medicine for treating endometriosis, and use this to treat endometrium in clinical trials The medicine for heterotopia is named as endometrium.

The medicine for treating endometriosis of the present invention is clinically named Neiyi Tablet, and the main function of the preparation is: removing blood stasis and dredging organs. Indications for endometriosis, symptoms include dysmenorrhea, pelvic pain, anal bulging, backache, constipation, dark tongue or ecchymosis, thin fur, thready or astringent pulse.

Among its medicinal prescriptions, this prescription uses raw rhubarb as the monarch drug, and rhubarb "passes away bitterly", "cleanses and clears the internal organs, pushes the old to bring about the new", "Compendium of Materia Medica" says that it has "lower stasis and blood closure..., broken symptoms and accumulation... ..., women's cold blood closed, lower abdomen pain, all old blood stagnation..., ventilating all qi, jumping blood vessels...." The merit of the minister's medicine, Biejia, "is the medicine for the liver meridian of Jueyin", and has the ability to "break the liver meridian and blood". The effect of "sickness in the abdomen and bad blood" is mainly used to treat "confined abdominal pain..., blood in the abdomen, low back pain..., women's meridians are blocked". Amber "eliminates blood stasis..., breaks knots...", "Zang Qi" says: "It is mixed with rhubarb and turtle shells... to produce bad blood, and blood in women's abdomen...". There is also turtle shell made with rice vinegar, which has a sour taste and enters the liver meridian, which has the functions of "removing symptoms and lumps..., breaking down qi..., and dispelling blood stasis". All the medicines are compatible, and the prescriptions are rigorous, and they have the effect of removing blood stasis and dredging the organs.

The medicine for treating endometriosis of the present invention can be made into various medicine forms such as tablet, powder, granule, oral liquid, injection, but the oral solid preparation is preferred. The above-mentioned medicines in various dosage forms can be prepared according to conventional methods in the field of pharmacy. When needed, one or more pharmaceutically acceptable adjuvants can also be added, and the adjuvants include conventional diluents, excipients, fillers, binders, wetting agents, absorption promoters, surfactants, etc. Agents, lubricants, stabilizers, etc., and flavoring agents, sweeteners, and pigments can also be added if necessary.

Through animal experiments, two endometriosis animal models of rats and rabbits were used to study the therapeutic effect and pharmacodynamic mechanism of Neiyi tablets. The results showed that: Neiyi tablets can reduce endometriosis It has obvious effect on the treatment of endometriosis; through the research on the pain model caused by glacial acetic acid, oxytocin and prostaglandin, it shows that Neiyi tablet has analgesic effect, and it is speculated that the analgesic mechanism of Neiyi tablet may be It is related to the reduction of prostaglandins, providing an experimental basis for its clinical relief of abdominal pain, dysmenorrhea and other symptoms in patients with endometriosis; using different inflammatory models to study, it shows that Neiyi Tablet has a significant anti-inflammatory effect, and Neiyi Tablet promotes local lesions. The study on the blood stasis model shows that Neiyi tablet has a significant effect of promoting blood circulation and removing blood stasis; in the test of the effect on hormone levels in normal rats, Neiyi tablet can reduce the level of estrogen and progesterone in normal rats. Experiments have shown that Neiyi tablets can treat endometriosis and alleviate its clinical symptoms through anti-inflammatory and analgesic effects, promoting blood circulation and removing blood stasis. Acute toxicity study results show that the maximum oral administration dose of Neiyi Tablets to mice is 63.6g/kg, which is 848 times the intended clinical dose for humans. In the 6-month long-term toxicity test of rats, compared with the blank control group, there was no significant difference in the general condition, weight gain, hematology and blood biochemical indicators, major organ coefficients, naked eye observation and histomorphological observation of the experimental animals compared with the blank control group Or abnormal, did not show obvious toxicity, indicating that Neiyi tablets are safe within the scope of clinical use. According to the observation of 114 cases of clinical trials, the total effective rate and total marked rate of Neiyi Tablets were 96.49% and 85.96%, respectively, which were equivalent to the curative effect of the Danazol group in the control group, but the adverse reactions were significantly less than those in the control group.

Due to the adoption of the above technical scheme, the present invention has the following advantages: 1. The medicinal formula of the present invention has obvious characteristics, and rhubarb is used to "walk down the bitterness", "cleanse the internal organs, push the old to bring about the new", and use the turtle shell to "break the symptoms and block the blood" The effect of amber, "removing blood stasis..., breaking knots..." is used to play the effect of removing blood stasis and dredging the internal organs, which further improves the curative effect; 3. Compared with the existing traditional Chinese medicine, the medicine of the present invention fully embodies the thought of "The internal classics" that "the internal organs are used for general use" and "the ones that are below should be exhausted". The whole prescription promotes blood circulation, removes blood stasis and clears blood stasis, and is specially used for the treatment of endometriosis. The clinical effect is remarkable without any adverse reactions.

Detailed ways

Embodiment 1, the preparation of the present invention's medicine for treating endometriosis

The medicine for treating endometriosis, the traditional Chinese medicine prescription is as follows: rhubarb 300g, turtle shell 300g, amber 150g.

The method for preparing the medicine for treating endometriosis comprises the following specific steps:

1) Weigh the medicinal materials according to the components and their weights in the above-mentioned traditional Chinese medicine prescription, and perform routine pretreatment;

2) Soak rhubarb plus 85% ethanol in 8 times amount for 1 hour, reflux extraction three times, each time for 1 hour, filter, combine the filtrate for 3 times, recover ethanol and concentrate under reduced pressure to a relative density of 1.08-1.10 (25°C) ;

3) Turtle shells were extracted four times with 10 times the amount of water, each time for 8 hours, filtered, combined filtrates, and concentrated under reduced pressure to a relative density of 1.08-1.10 (25°C).

4) Combine the concentrates of steps 2) and 3), add amber, mix well, dry in vacuum below 75°C and pulverize into fine powder,

5) Add appropriate amount of auxiliary materials to the fine powder in step 4), mix well, make granules, press into 1000 tablets, coat with film, and obtain the medicine for treating endometriosis, and use this to treat endometrium in clinical trials The medicine for heterotopia is named as endometrium.

The finished Neiyi tablet made in this example is a film-coated tablet, which is khaki to light brown after removing the film coating; it tastes salty and slightly bitter.

Test according to thin-layer chromatography (Appendix VI B of Chinese Pharmacopoeia in 2005), draw the solution of the test product and 1 μl of the contrast medicinal material solution, respectively spot on the same silica gel HSG thin-layer plate, and use benzene: methanol: glacial acetic acid (79 : 14: 7) as developer, develop, dry, and inspect under ultraviolet light (365nm). The chromatogram of the test product shows fluorescent spots of the same color at the position corresponding to the chromatogram of the control medicinal material.

Adopt according to thin-layer chromatography ("Chinese Pharmacopoeia" edition in 2005 an appendix VI B), the quality index of the finished product of endometriosis sheet that present embodiment is made is checked, draw need testing solution, each 1 μ l of contrast medicinal material solution, point respectively On the same silica gel HSG thin-layer plate, develop with benzene:methanol:glacial acetic acid (79:14:7) as developing solvent, dry in the air, and observe under ultraviolet light (365nm). The test product chromatogram shows fluorescent spots of the same color at the position corresponding to the contrast medicinal material chromatogram, and other indicators meet the relevant regulations under the tablet item ("Chinese Pharmacopoeia" 2005 edition one appendix I J)

Using high performance liquid chromatography (Appendix VI D of "Chinese Pharmacopoeia" 2005 edition), the main active ingredients of the finished products of Neiyi Tablets made in this embodiment are determined, and each tablet contains rhubarb and chrysophanol (C ₁₅ H ₁₀ O ₄ ), emodin (C ₁₅ H ₁₀ O ₅ ) should not be less than 1.50mg.

In this example, in order to verify the effect of the drug, a drug preparation for the experiment was prepared, and the preparation method was the same as above, except that a fixed ratio of raw materials was used.

Embodiment 2, the experiment of the influence of the medicine of the present invention for treating endometriosis on rat endometriosis model

Adopt the medicine for the treatment of endometriosis prepared by embodiment 1 (clinical experiment is named as endometriosis sheet), carry out animal experiment to rat endometriosis, by contrast experiment, investigate medicine of the present invention to endometrium Therapeutic effects in atopic rats. The specific experimental process is as follows:

(1) Methods: Wistar rats were used to establish endometriosis animal models by autologous endometrial pelvic transplantation. 0.1ml/kg of estradiol benzoate was intramuscularly injected 2 days before modeling, and intraperitoneally injected with pentobarbital sodium 30mg/kg for 2 days. From the right side of the uterus to 2cm away from the ovary, a section of uterus about 2cm in the middle was taken after ligation, the endometrium was stripped from the muscle layer, cut into a piece of endometrium about 5mm×5mm in size, sutured to the inner wall of the abdominal cavity, and the abdominal cavity was closed after the suturing was completed. Rats in the sham-operated group were laparotomed, the endometrium was not stripped, and the same volume of abdominal fat was taken and sutured to the abdominal wall. Postoperative intramuscular injection of penicillin sodium 80,000 u/kg, once a day, for 3 consecutive days, and intramuscular injection of estradiol benzoate 0.1ml/kg on the second day after operation, once every 3 days, 3 times in total. To promote the growth of its transplanted intima, routine feeding. Three weeks after the operation, the second laparotomy was performed, and the success of the modeling was based on the fact that a hillock-like or transparent effusion was seen under the ectopic endometrium. Rats with a volume of ectopic endometrium ranging from 20 to 70 mm ³ were selected and grouped according to the volume. The average volume of ectopic intima in each group was similar.

(2) Grouping: In addition to the normal group and the sham operation group, the modeled rats were divided into a model group, a large, medium and small dose group of Neiyi tablet, and a positive drug danazol group, with 10 rats in each group. After 4 weeks of modeling, the Neiyi Tablet group was fed with 1.78 g crude drug/kg, 0.89 g crude drug/kg, and 0.45 g crude drug/kg of large, medium, and small doses of Neiyi Tablet suspension, once a day. The danazol group was fed with 0.20 g/kg of danazol, and the normal group, the sham operation group, and the model group were fed with equal-volume distilled water (10ml/kg), once a day, for 30 days in a row. kg anesthetized, blood was collected from the abdominal aorta, and then organs such as ectopic endometrium, eutopic endometrium, and ovary were collected. Observation: (1) Measure the length, width and height of the ectopic endometrium with a vernier caliper, and calculate the volume of the ectopic focus; (2) Observe the pathological changes of the ectopic endometrium, eutopic endometrium, and ovarian tissue morphology under the HE staining light microscope (3) Weigh the left uterus, left ovary, and thymus with an electronic balance, and calculate the indexes of each organ; (4) LBY-N6B self-cleaning rotary viscometer measures blood rheology indicators, including whole blood specific viscosity ( Low cut, medium cut, high cut), plasma specific viscosity, hematocrit, erythrocyte sedimentation index, erythrocyte aggregation index, erythrocyte sedimentation equation K value, erythrocyte rigidity index, erythrocyte deformation index, erythrocyte electrophoresis index.

(3) Results:

The effect of A on the volume of ectopic intima

The results showed that before administration, the volume of ectopic intima in rats of Neiyi Tablet group and Danazol group was not significantly different from that of the model group (P>0.05). After 30 days, the volume of ectopic intima in the model group was 42.8±21.5mm ³ , and the volume of ectopic intima was slightly smaller than that before 30 days. The volume of ectopic intima in the large, medium and small dose groups of Neiyi Tablets and Danazol group decreased significantly after 30 days of administration, compared with the model group, there was a very significant difference (P<0.01). Compared with before administration, the volume of ectopic intima also decreased significantly, with a very significant difference (P<0.01). See Table 1.

Table 1 The effect of Neiyi tablet on the volume of ectopic endometrium in endometriosis model rats (X±s, mm ³ )

Compared with the model group *P<0.05, **P<0.01, compared with before treatment ^▲ P<0.05, ^▲▲ P<0.01

B Histopathological examination

The results showed that more endometrial stroma and glands were seen in the pathological tissue of ectopic endometrium in rats in the model group. Compared with the model group, the ectopic intima-interstitial area and the number of endometrial glands in the rats in the endometriosis group were significantly reduced, and the reduction in the high-dose group was the most significant. The number of ectopic endometrial stroma and glands in the Danazol group was similar to that in the Neiyi tablet medium dose group. The eutopic endometrium in each group had normal endometrial structure, see stroma and glands; the ovary had normal ovarian structure, see growing follicles and mature follicles at all levels.

Effect of C on viscera index

The results showed that the uterine index and ovary index of the rats in the model group increased significantly, compared with the normal group, there was a significant difference (P<0.05). Endometriosis tends to reduce uterine index and thymus index. The ovarian index in the danazol group decreased significantly, compared with the model group, there was a very significant difference (P<0.01), and the thymus index tended to decrease. See Table 2.

Table 2 Influence of Neiyi Tablets on the Visceral Index of Endometriosis Model Rats (X ± s, mg/100g)

Compared with the model group *P<0.05, **P<0.01

Effect of D on hemorheology

The results showed that the various indicators of hemorheology in the model group: whole blood low shear, whole blood middle shear, whole blood high shear, plasma specific viscosity, hematocrit, erythrocyte sedimentation rate, whole blood reduction low shear, whole blood reduction middle cut, Whole blood reduced high shear, erythrocyte aggregation index, erythrocyte deformation index, erythrocyte rigidity index, hemocyte electrophoresis index, erythrocyte sedimentation equation K value and other indicators had no significant difference compared with the normal group (P>0.05). Compared with the model group or normal group, there was no significant difference in hemorheology indexes between Neiyitablet group and Danazol group (P>0.05). See Table 3.

Table 3 Effect of Neiyi Tablets on Hemorheology in Endometriosis Model Rats (X ± s)

Compared with the model group *P<0.05, **P<0.01

Embodiment 3, the experiment of the influence of the medicine of the present invention for treating endometriosis on domestic rabbit endometriosis model

Adopt the medicine for the treatment of endometriosis prepared by embodiment 1 (clinical experiment is named after endometriosis), to the therapeutic effect test of endometriosis model rabbit, by contrast experiment, investigate medicine of the present invention to endometriosis Therapeutic effect of biting rabbits. The specific experimental process is as follows:

(1) Methods: 80 New Zealand rabbits were used to establish animal models of endometriosis by autologous endometrial pelvic transplantation. Intramuscularly inject estradiol benzoate (0.04ml/kg) 2 days before modeling, for a total of 2 days, inject pentobarbital sodium 30mg/kg intraperitoneally, perform laparotomy under sterile conditions after anesthesia, and ligate the mesentery For blood vessels, separate the right uterus to 2cm away from the ovary. After ligation, take a section of uterus about 2cm in the middle, peel off the endometrium from the muscle layer, cut into two pieces of endometrium about 5mm×5mm in size, and suture them on the left and right abdominal walls. Close the abdominal cavity after completion. Rabbits in the sham operation group underwent laparotomy without peeling off the endometrium, and the same volume of fat from the abdominal cavity was sutured to the left and right inner walls of the abdominal wall. Postoperative intramuscular injection of penicillin sodium (40,000 u/kg), once a day, for 3 consecutive days. The intramuscular injection of estradiol benzoate (0.04ml/kg) was started on the 2nd day after the operation, once every 4 days, a total of 3 times to promote the growth of the transplanted intima, and they were fed normally. 4 weeks after the operation, the second laparotomy was performed. The successful modeling was based on the fact that hillocks or transparent effusions were seen under the ectopic endometrium. The ectopic endometrium volume on one side was selected from ³ rabbits with a volume of 200-700 mm. Discarded and grouped according to volume, so that the average volume of ectopic intima in each group is similar.

(2) Grouping: In addition to the normal group and the sham operation group, the rabbits with successful modeling were divided into a model control group, a large, medium and low dose group of Neiyi tablet, and a positive drug danazol group, with 8 rabbits in each group. At 5 weeks after operation, the Neiyi Tablet group was given large, medium and small doses of suspension of Neiyi Tablet 1.20 g crude drug/kg, 0.60 g crude drug/kg, 0.30 g crude drug/kg, once a day. The danazol group was fed with 0.11g/kg of danazol, and the normal group, the sham operation group, and the model group were fed with equal volume of distilled water (1ml/kg), once a day, for 30 consecutive days. kg anesthetized, blood was collected from the heart, and then the ectopic endometrium, pituitary and other organs were collected by laparotomy. Observation (1) Measure the length, width and height of the ectopic endometrium with a vernier caliper, and calculate the volume of the ectopic focus; (2) Observe the pathological changes of the ectopic endometrium and pituitary tissue morphology under the HE staining light microscope; (3) Electronic balance Weigh the left uterus, left ovary, thymus, and calculate the index of each organ; (4) LBY-N6B self-cleaning rotary viscometer to measure the hemorheology index, including whole blood specific viscosity (low cut, middle cut, high shear), plasma specific viscosity. (5) Determination of plasma TXB ₂ by ELX800 microplate reader.

(3) Results:

The effect of A on the volume of ectopic intima

The results showed that before administration, there was no significant difference in the volume of ectopic intima in rabbits in the large, medium and low dose groups of Neiyi Tablets and Danazol group compared with the model group (P>0.05). After 30 days, the volume of ectopic intima in the model group was 723.8±265.7m ³ , which was significantly larger than that before 30 days. The volume of ectopic intima in the Neiyi tablet high-dose group and the danazol group was significantly reduced, compared with the model group, there was a very significant difference (P<0.01); compared with before administration, it was also significantly reduced, with a very significant difference Difference (P<0.01). See Table 4.

Table 4 The influence of endometriosis film on the volume of ectopic endometrium in rabbit endometriosis model (X±s, mm ³ )

Compared with the model group *P<0.05, **P<0.01, compared with before treatment ^▲ P<0.05, ^▲▲ P<0.01

B Histopathological examination

The results showed that more endometrial stroma and glands were seen in the pathological tissue of ectopic endometrium in rabbits in the model group. Compared with the model group, the ectopic endometrium interstitial area and the number of endometrial glands in the rabbits in the endometriosis group were significantly reduced, and the reduction in the high-dose group was the most significant. The number of ectopic endometrial stroma and glands in the Danazol group was similar to that in the Neiyi tablet medium dose group.

In the normal group, the proportions of eosinophils, basophils and chromophobe cells in the anterior pituitary were normal, and no cell degeneration and necrosis were observed. The model group, Neiyi high-dose group were similar to the normal group, and the cell density under the microscope was not significantly sparse, while the medium-dose group, low-dose and danazol groups were significantly less than the normal group, and the cell density was significantly sparse under the microscope.

Effect of C on viscera index

The results showed that the rabbit uterus index, ovary index, and thymus index in the model group had no significant changes compared with the normal group, and there was no significant difference in the uterine index, ovary index, and thymus index of the endometrium group compared with the model group or the normal group (P> 0.05). The uterine index in the danazol group was significantly increased, and the thymus index was significantly reduced, compared with the model group, there was a significant difference (P<0.05, P<0.01). See Table 5.

Table 5 The influence of endometriosis tablet on the organ index of rabbit model of endometriosis (X ± s, mg/100g)

Compared with the model group *P<0.05, **P<0.01

Effect of D on hemorheology

The results showed that the hemorrheology indexes of the model group: whole blood low shear, whole blood middle shear, whole blood high shear, plasma specific viscosity, had no significant difference compared with the normal group. Compared with the model group or normal group, there was no significant difference in hemorheology indexes between Neiyitablet group and Danazol group (P>0.05). See Table 6.

Table 6 Effect of Neiyi Tablets on Hemorheology of Endometriosis Model Rabbits (X±s)

Compared with the model group *P<0.05, **P<0.01

Effect of E on plasma TXB ₂

The results showed that the TXB ₂ of rats in the model group increased significantly, which was 691.62±219.08pg/ml, and the TXB ₂ of the high-dose Neiyi tablet group decreased significantly, compared with the model group, there was a significant difference (P<0.05). See Table 7.

Table 7 The influence of endometriosis sheet on rabbit TXB ₂ of endometriosis model (X ± s, pg/ml)

Compared with the model group *P<0.05, **P<0.01

Embodiment 4, the drug of the present invention for treating endometriosis is tested against xylene-induced ear swelling in mice

Adopt the medicine of the treatment endometriosis prepared in embodiment 1 (clinical experiment is called as Neiyi tablet), anti-xylene causes mouse ear swelling effect test, by control experiment, investigates medicine of the present invention to mouse ear swelling inflammation therapeutic effect. The specific experimental process is as follows:

60 female mice were taken, weighed, and randomly grouped. Divided into blank control group, Neiyi tablet large, medium and small dose groups, positive drug aspirin group, danazol group. The Neiyi Tablet group was fed with large, medium and small doses of Neiyi Tablet suspension of 1.53g crude drug/kg, 0.76g crude drug/kg, and 0.38g crude drug/kg, the aspirin group was fed with aspirin 75mg/kg, and the danazol group was fed with Danazol 0.20g/kg was taken, and the blank control group was given equal volume of distilled water 10ml/kg once a day for 3 consecutive days. One hour after the last administration or distilled water, the right ear of each mouse was coated with 50 μl of xylene per mouse, and sacrificed 20 minutes later. The same parts of both ears were cut off with a hole punch with a diameter of 7 mm, and weighed separately with a precision electronic balance. The difference between the weights of the left and right ear pieces was used as the degree of swelling, and the degree of swelling of each mouse ear and the percentage of inhibition of swelling in each group were calculated.

Formula: swelling degree = weight of right ear piece - weight of left ear piece

The results showed that the right ear of the mice in the blank control group appeared red and swollen, and the ear swelling degree was as high as 19.6±3.4mg. The ear swelling degree of the mice in the large, medium and small dose groups of Neiyi tablet was significantly reduced, which was significantly higher than that of the blank control group. Difference (P<0.01), and there is a dose-effect relationship. The swelling inhibition rates were 31.1%, 30.1%, and 29.6%, respectively. The degree of ear swelling of the mice in the aspirin group and the danazol group was also reduced, and compared with the blank control group, there was a very significant difference (P<0.01). See Table 8.

Table 8 Neiyi Tablet Anti-Xylene Causes Mice Ear Swelling Effect (X ± s)

Compared with blank control group *P<0.05, **P<0.01

Embodiment 5, the anti-rat cotton ball granuloma effect test of the medicine of the present invention for treating endometriosis

Adopt the medicine of the treatment endometriosis prepared in embodiment 1 (clinical experiment is named as Neiyi tablet), anti-rat cotton ball granuloma effect experiment, by contrast experiment, investigate medicine of the present invention to cotton ball granuloma rat therapeutic effect. The specific experimental process is as follows:

60 female Wistar rats were randomly divided into blank control group, large, medium and small dose groups of Neiyi tablet, positive drug prednisolone acetate group and danazol group, 10 rats in each group, intraperitoneal injection of pentobarbital Anesthetized with sodium 30 mg/kg, cut off the rat hair on the back of the rat, sterilized with 75% alcohol, cut a 1.0 cm long incision, and implanted 30 mg cotton ball (sterilized by high pressure) from the incision under the back skin with ophthalmic forceps, and then sutured the skin , starting from the day of operation, the Neiyi Tablets group was administered intragastrically with large, medium, and small doses of suspensions of 1.78 g crude drug/kg, 0.89 g crude drug/kg, and 0.45 g crude drug/kg respectively; Take prednisolone acetate 9.0mg/kg, the danazol group was given danazol 0.20g/kg, and the blank control group was given equal volume of distilled water (10ml/kg), once a day for 10 consecutive days. 24 hours after the last administration, cut the incision, take out the cotton ball together with the connective tissue, remove the adipose tissue, and dry it in an oven at a constant temperature of 70°C for 48 hours, then weigh it with an electronic balance, and subtract the weight of the cotton ball from the weight obtained. That is, the weight of the granuloma was obtained.

The results showed that the dry weight of the rat cotton ball granuloma in the blank control group was 70.1 ± 16.5 mg, and the dry weight of the rat cotton ball granuloma in the large and medium dose groups of Neiyi tablets was significantly reduced, and there was a very significant difference compared with the blank control group (P <0.01), and there is a dose-effect relationship. The dry weight of the cotton ball granuloma in the prednisolone acetate group and the danazol group was also significantly reduced, and compared with the blank control group, there was a very significant difference (P<0.01). See Table 9.

Table 9 Anti-rat Cotton Ball Granuloma Effect of Neiyi Tablets (X ± s, mg)

Compared with blank control group *P<0.05, **P<0.01

Embodiment 6, the medicine of the present invention for the treatment of endometriosis is tested against the writhing effect of glacial acetic acid on mice

Adopt the medicine for the treatment of endometriosis prepared in embodiment 1 (clinical experiment is named as Neiyi tablet), anti-glacial acetic acid causes mouse writhing effect test, by control experiment, investigates medicine of the present invention anti-glacial acetic acid causes mouse Therapeutic effect of writhing pain. The specific experimental process is as follows:

Take 60 female mice and divide them into blank control group, large, medium and small dose groups of Neiyi tablets, positive drug aspirin group, and Danazol group. The Neiyi Tablet group received Neiyi Tablet large, medium and small dose suspensions of 1.53g crude drug/kg, 0.76g crude drug/kg, and 0.38g crude drug/kg respectively; Danazol 0.20g/kg, and the blank control group were intragastrically administered equal volume of distilled water (10ml/kg), once a day for 3 consecutive days. One hour after the last administration, mice were intraperitoneally injected with 0.2 ml of 0.9% glacial acetic acid solution, and the number of writhing times of mice in each group was observed within 20 minutes.

The results showed that the writhing of the mice in the blank control group showed abdominal depression, contraction, arching of the back, stretching of the limbs, and opisthotonos, and the number of writhing times was as high as 53.3±16.5 times. The number of writhing times of the rats was significantly reduced, which was significantly different from that of the blank control group (P<0.01). The times of writhing of the mice in the aspirin group and the danazol group were also significantly reduced, compared with the blank control group, there were significant differences (P<0.01, P<0.05). See Table 10.

Table 10 Neiyi tablets anti-glacial acetic acid-induced writhing effect on mice (X ± s, times)

Compared with blank control group *P<0.05, **P<0.01

Embodiment 7, the drug of the present invention for the treatment of endometriosis The test of anti-oxytocin-induced writhing effect in mice

Adopt the medicine for the treatment of endometriosis prepared in embodiment 1 (clinical experiment is named after the endometriosis tablet), anti-oxytocin causes mouse body writhing effect test, by control experiment, investigates medicine of the present invention anti-oxytocin-induced mouse Therapeutic effect of writhing pain. The specific experimental process is as follows:

Take 60 female mice and divide them into blank control group, Neiyi tablet large, medium and low dose groups, aspirin group and danazol group. All groups were given diethylstilbestrol 2mg/kg once a day for 12 consecutive days. On the 10th day, the Neiyi Tablet group was fed with large, medium and small doses of suspensions of 1.53 g crude drug/kg, 0.76 g crude drug/kg, and 0.38 g crude drug/kg respectively; the aspirin group was fed with aspirin 75 mg/kg, danazol The control group was fed with danazol 0.20g/kg, and the blank control group was fed with equal volume of distilled water (10ml/kg), once a day for 3 consecutive days. One hour after the last administration, the mice were intraperitoneally injected with 20u/kg oxytocin solution, and the writhing times of the mice in each group were observed within 30 minutes.

The results showed that the writhing of the mice in the blank control group showed abdominal contraction, the front legs were stretched forward, and the hind legs were stretched back. The number of writhing times was as high as 29.4±9.3 times. , Compared with the blank control group, there was a very significant difference (P<0.01). The times of writhing of the mice in the aspirin group and the danazol group were also significantly reduced, compared with the blank control group, there was a very significant difference (P<0.01). See Table 11.

Table 11 Neiyi tablet anti-oxytocin-induced writhing effect on mice (X ± s, times)

Compared with blank control group *P<0.05, **P<0.01

Example 8, the present invention's drug for treating endometriosis is tested against prostaglandin-induced writhing effect in mice

Adopt the medicine for the treatment of endometriosis prepared in embodiment 1 (clinical experiment is named as Neiyi Tablet), anti-prostaglandin-induced mouse writhing effect test, by control experiment, investigate the medicine of the present invention anti-prostaglandin-induced mouse Therapeutic effect of writhing pain. The specific experimental process is as follows:

Take 60 female mice and divide them into blank control group, Neiyi tablet large, medium and low dose groups, aspirin group and danazol group. The Neiyi Tablet group was fed with large, medium and small doses of Neiyi Tablet suspension of 1.53g crude drug/kg, 0.76g crude drug/kg, and 0.38g crude drug/kg respectively; the aspirin group was fed with aspirin 75mg/kg, Danat The azole group was fed with danazol 0.20g/kg, and the blank control group was fed with equal volume of distilled water (10ml/kg), once a day for 3 consecutive days. One hour after the last administration, mice were intraperitoneally injected with 0.2% PGF _2α 10ml/kg, and the number of writhing within 40min was observed.

The results showed that the writhing times of the mice in the blank control group were contracted and concave in the abdomen, the buttocks were raised, and the number of writhing times was as high as 14.0±6.2 times. Compared with the blank control group, there was a very significant difference (P<0.01). The times of writhing of the mice in the aspirin group and the danazol group were also significantly reduced, compared with the blank control group, there was a very significant difference (P<0.01). See Table 12.

Table 12 The anti-prostaglandin-induced mouse writhing effect of Neiyi tablet (X ± s, times)

Compared with blank control group *P<0.05, **P<0.01

Example 9, the experiment of the effect of the medicine for treating endometriosis of the present invention on the effect of promoting blood circulation and removing blood stasis

Using the medicine for treating endometriosis prepared in Example 1 (named Neiyi Tablet for clinical experiments), animal experiments were carried out on the effect of promoting blood circulation and removing blood stasis, and by controlled experiments, the effect of the medicine of the present invention on promoting blood circulation and removing blood stasis was investigated. The specific experimental process is as follows:

60 SD rats, weighing 180-220 g, were randomly divided into normal group, model group, large, medium and small dose groups of Neiyi Tablets, and Danshen Tablets group. The Neiyi Tablet group was fed with large, medium and small doses of Neiyi Tablet suspension of 1.78g crude drug/kg, 0.89g crude drug/kg, and 0.45g crude drug/kg respectively, and the Compound Danshen Tablet group was fed with Compound Danshen Tablet 0.54g/kg , the normal group and the model group were fed with equal volume of distilled water (10ml/kg), once a day, for 15 consecutive days. On the 14th day, except the normal group, the rats in the other groups were subcutaneously injected with Adr, 10 μg/kg each time, twice in total with an interval of 4 hours. The normal group was subcutaneously injected with an equal volume of normal saline. After treatment, they were fasted without water overnight. One hour after the last administration or distilled water, except the normal group, the rats in all other groups were subcutaneously injected with Adr 10 μg/kg, and then immersed in ice water at 4°C for 5 minutes after 20 minutes. The normal group was subcutaneously injected with an equal volume of normal saline without immersion in ice water. Each rat was anesthetized with 1 g/kg of wulai sugar, and 4 ml of blood was collected from the abdominal aorta.

Immediately inject 1ml of blood into the rotating ring according to Chandler's extracorporeal method, seal it quickly, place it on a thrombosis instrument and rotate it for 15 minutes, pour out the thrombus, blot the fresh blood on the surface with filter paper, weigh the wet weight of the thrombus, and measure the length of the thrombus; sterilize the thrombus in hot air The thrombus was dried in a cabinet at 120°C for 60 minutes, and the dry weight of the thrombus was measured. 3ml blood viscosity was measured with LBY-N6B self-cleaning rotary viscometer.

The results showed that the thrombus length, thrombus wet weight and dry weight of rats in the model group were significantly increased, which was significantly different from the normal group (P<0.01), indicating that the blood stasis model was successful. The wet weight and dry weight of thrombus in the large and medium dose groups of Neiyi Tablets decreased, which was significantly different from that in the model group (P<0.05). The wet weight and dry weight of thrombus in the Danshen group were decreased, which was significantly different from that in the model group (P<0.05). See Table 13.

Table 13 Influence of Neiyi tablet on ice water adrenaline-induced rat thrombosis (X ± s)

Compared with the model group *P<0.05, **P<0.01

The low-shear, middle-shear and high-shear of rats in the model group were significantly increased, which was significantly different from that of the normal group (P<0.05). The low-shear of whole blood of the rats in the large, medium and low dose groups of Neiyi tablets decreased, which was significantly different from that of the model group (P<0.05). The low-shear, middle-shear and high-shear of whole blood in the Danshen group all decreased, and there was a significant difference compared with the model group (P<0.01). Compared with the model group, there was no significant difference (P>0.05) in the plasma specific viscosity of the Neiyitablet group and the Danshen group. See Table 14.

Table 14 Effect of Neiyi Tablets on hemorheology in ice water epinephrine-induced blood stasis model rats (X ± s)

Compared with the model group *P<0.05, **P<0.01

Embodiment 10, the influence experiment of the medicine of the present invention for treating endometriosis on normal rat hormone level

Adopt the medicine for the treatment of endometriosis prepared by embodiment 1 (clinical experiment is named as Neiyi tablet), carry out animal experiment to the influence of normal rat hormone level, by contrast experiment, investigate medicine of the present invention to normal rat hormone level of impact. The specific experimental process is as follows:

Fifty female Wistar rats were randomly divided into blank control group, large, medium and small dose groups of Neiyi Tablet, and positive drug danazol group, with 10 rats in each group. The Neiyi Tablet group was fed with Neiyi Tablet large, medium and small dose suspensions of 1.78g crude drug/kg, 0.89g crude drug/kg, and 045g crude drug/kg respectively; the Danazol group was fed with Danazol 0.20g/kg, and the blank control The rats in the group were fed with equal volume of distilled water (10ml/kg), once a day, for 30 consecutive days. After 30 days, the rats were anesthetized with 1 g/kg wulai sugar, blood was drawn from the abdominal aorta, and serum E2, P, and LH were measured by radioimmunoassay.

The results showed that the levels of estrogen and progesterone in rats in the middle-dose Neiyi Tablet group were significantly lower than those in the normal group (P<0.05). The levels of estrogen and progesterone in rats in the danazol group were significantly lower than those in the normal group (P<0.01). See Table 15.

The impact of table 15 on normal rat hormone levels (X ± s, pg/ml)

Compared with the model group, *P<0.05, **P<0.01, because the variances are not homogeneous in the table, the statistical results were obtained by correcting the homogeneity of variances by rank method and open method.

Example 11, Clinical Trial of Neiyi Tablets for Treating Endometriosis

1. Selection of clinical trial cases:

According to "Guidelines for Clinical Research of New Chinese Medicines" 5th edition in 2002; 114 cases of endometriosis who met the diagnostic criteria were selected (cases were selected according to the ratio of 1:1 between the treatment group and the control group).

2. Selection of Drugs for Clinical Trials

(1) Test drug: the Neiyi Tablet preparation prepared in Example 1 above, specification: 0.3 g/tablet.

(2) Control drug: danazol, specification: 100 mg/grain.

3. Methods of Clinical Trials

(1) Random grouping of cases: each block is numbered according to the sequence of 50% of mild disease and 50% of moderate disease, and randomly divided into treatment group and control group according to the ratio of 1:1 between treatment group and control group.

(2) The clinical trial observation method adopts the double-blind observation method.

(3) Medication method and dosage:

Medication for the treatment group: Neiyi Tablets, orally, 2 tablets each time, three times a day.

Medication for the control group: danazol, orally, 2 capsules each time, three times a day.

Course of treatment: The treatment group and the control group both take medicine for 3 months as a course of treatment, and the observation time is a course of treatment.

4. Observation and evaluation of clinical trial drug efficacy:

1. Clinical recovery: symptoms basically disappeared; positive results disappeared after treatment; infertility patients conceived during treatment and follow-up.

2. Significantly effective: the symptoms basically disappear, and the symptom score before and after treatment is reduced by more than 60% or the degree of symptoms drops by two levels; Period contrast); infertile patients were able to conceive although local signs existed.

3. Effective: Symptom relief before and after treatment, the symptom score decreased by more than 30%; the symptom score did not increase (comparison of menstrual cycle examination at the same period; B-ultrasound examination before and after treatment at the same period of comparison); symptoms did not increase within 3 months of drug withdrawal.

4. Ineffective: before and after treatment, the scores of symptoms and signs decrease by less than 30% or the scores increase.

5. Conclusions of clinical trials:

The clinical trial of a course of treatment of Neiyi Tablets and matched group oral danazol through the treatment group, the results are shown in Table 15:

Table 15: Results of clinical trials of Neiyi Tablets in the treatment of endometriosis

Claims (4)

1. treat endometriotic medicine for one kind, it is characterized in that it is to be made by the raw material that contains following ratio of weight and number:

Radix Et Rhizoma Rhei 2.8～3.2 Carapax Trionycis 2.8～3.2 Succinum 1.3～1.7

2. the endometriotic medicine of a kind of treatment as claimed in claim 1, it is characterized in that: it is to be made by the raw material that contains following ratio of weight and number:

Radix Et Rhizoma Rhei 2.9～3.1 Carapax Trionycis 2.9～3.1 Succinum 1.4～1.6

3. the endometriotic medicine of a kind of treatment as claimed in claim 2, it is characterized in that: it is to be made by the raw material that contains following ratio of weight and number:

Radix Et Rhizoma Rhei 3 Carapax Trionycis 3 Succinum 1.5

4. method of producing the endometriotic medicine of the described treatment of claim 3 may further comprise the steps:

1) take by weighing the raw material of following ratio of weight and number:

Radix Et Rhizoma Rhei 2.8～3.2 Carapax Trionycis 2.8～3.2 Succinum 1.3～1.7

2) Radix Et Rhizoma Rhei is added 85% 8 times amount soak with ethanol 1 hour, reflux, extract, three times, each 1 hour, filter, merges 3 times filtrate, being evaporated to relative density behind the recovery ethanol is 1.08-1.10 (25 ℃);

3) Carapax Trionycis, Concha Ostreae, Typhonium flagelliforme (Lodd.) Blume, Radix Rhapontici add 10 times of water gagings extractions four times, and each 8 hours, filter, merging filtrate is evaporated to relative density 1.08-1.10 (25 ℃).

4) with step 2) and 3) concentrated solution merge, add succinum, mixing is ground into fine powder behind the vacuum drying below 75 ℃,

5) fine powder with step 4) adds appropriate amount of auxiliary materials, and mixing is made granule, is pressed into 1000, and the bag film-coat promptly gets the medicine for the treatment of endometriosis, and the drug nomenclature with this treatment endometriosis during clinical experiment is interior different.