WO2021208280A1 - Application of dried fly larva in preparation of drugs for preventing and treating breast hyperplasia - Google Patents

Abstract

Provided is a dried fly larva and an application of same in the preparation of drugs for preventing and treating breast hyperplasia and for improving estrogen levels.

Description

Application of Wu Gu Chong in the preparation of medicines for preventing and treating mammary gland hyperplasia Technical field

The invention belongs to the technical field of medicine, and particularly relates to the application of the worms in the preparation of drugs for preventing and treating breast hyperplasia.

Background technique

Hyperplasia of mammary glands (HMG) is a common chronic proliferative disease of mammary gland structure disorder in women, which can cause abnormalities in the structure and tissue morphology of the mammary glands, manifested by different degrees of hyperplasia and incomplete involution of the mammary glands and interstitium. . The incidence of breast hyperplasia in women’s breast tissue is 30% to 60%. It is a common disease in women. It has a high incidence. With the deepening of research on it, scholars have proposed that breast hyperplasia is related to breast cancer. , And the rate of malignant transformation is gradually increasing. In recent years, attention to women's health, that is, the prevention and treatment of breast hyperplasia, has become a hot spot in the medical field.

At present, there is no clinically applicable drug or program for the treatment of hyperplasia of mammary glands. Western medicine usually uses hormone preparations such as androgen, progesterone, tamoxifen, bromocriptine, danazol, and iodine preparations for the treatment of HMG. However, due to its curative effect has certain side effects and is prone to relapse, many scholars are opposed to it, and the compliance of patients is relatively poor. Hyperplasia of mammary glands is the dominant disease in traditional Chinese medicine. In recent years, the curative effect of traditional Chinese medicine in the treatment of hyperplasia of mammary glands has been affirmed by more and more doctors and patients. It has become an important treatment method due to its significant advantages such as small side effects. Traditional Chinese medicine can effectively treat breast hyperplasia with little side effects.

The dried larva (maggot) of the insect Diptera Chrysomya megacephala (also known as Chrysomya megacephala) and its close relatives (such as the housefly Musca domestica ) is a traditional Chinese medicine blindly called "five worms." , Also known as Water Fairy and Luo Fairy, commonly known as "maggots". According to the records of "Materia Medica Seeking Source" and "Materia Medica Biandu", the grain insects are cold in nature and non-toxic, enter the spleen and stomach meridian. After drying and grinding, they can be applied and applied for external use. It can treat ecthyma, lip furuncle and carbuncle and other infectious surgery disease. Wu Gu Chong was first published in the 40th volume of the Insect Department of Compendium of Materia Medica. It has the functions of clearing heat and eliminating stagnation, and has the pharmacological effects of treating malnutrition, abdominal distension and malnutrition.

technical problem

At present, there is no research report on Wu Gu Chong's treatment of breast hyperplasia.

Technical solutions

The purpose of the present invention is to provide a new use of grain insects for the prevention and treatment of mammary gland hyperplasia.

More specifically, it provides the application of Wu Gu Chong in the preparation of medicines for preventing and treating breast hyperplasia.

More specifically, it is the application of Wu Gu Chong in the medicine for reducing the prolactin hormone level of mammary gland hyperplasia.

As a preference, the use of the extracts of Chinese grain insects in the preparation of medicines for preventing and treating breast hyperplasia.

More preferably, the grain insect extract is an alcohol-water double extract of grain insect.

Application of Wu Gu Chong in the treatment of uterine thickening drugs caused by increased levels of exogenous estrogen.

Application of Wu Gu Chong in the treatment of nipple nodules caused by increased levels of exogenous estrogen.

The invention also provides a medicine for preventing and treating hyperplasia of mammary glands.

Preferably, the medicament may also contain pharmaceutically acceptable conventional auxiliary materials, such as disintegrants, lubricants, emulsifiers, adhesives, etc., which are directly or indirectly added to the pharmacy by conventional methods of traditional Chinese medicine preparations through conventional processing. Acceptable excipients.

Preferably, the clinical preparation of the drug is granules, capsules, tablets, pills or oral liquids, which are easy to take, highly targeted, and have obvious curative effects.

Beneficial effect

The present invention has the following beneficial effects:

Provide new uses of grain insects and expand the scope of use of grain insects;

Provides new prevention and treatment ideas for breast hyperplasia;

The treatment of mammary gland hyperplasia with Wu Gu Chong has obvious curative effect, direct access to the disease, and small side effects.

Description of the drawings

Figure 1 is a bar graph of the weight changes of female rats before modeling, after modeling, and after administration;

Figure 2 is a bar graph showing changes in the height of the nipple of SD female rats before modeling, after modeling, and after administration;

Figure 3 is a bar graph showing changes in the nipple diameter of SD female rats before modeling, after modeling, and after administration;

Figure 4 is a bar graph showing the effect of Wu Gu Chong on the uterine index of SD rats;

Figure 5 is a bar graph showing the effect of Wu Gu Chong on the levels of serum estradiol (E2) and prolactin (PRL) in rats.

The best mode of the present invention

Example 1

Zhou, a 48-year-old female patient from Lincheng Township, Changxing County, Zhejiang Province, had undergone breast lobular hyperplasia resection, and her breasts were swollen like a woman's breasts during breastfeeding. The pain was severe and her hands could not be touched. On the recommendation of a friend, she came to the Chinese Medicine Clinic. Purchased Wu Gu insect powder for oral treatment, twice a day, 3 grams each time. After taking it for three months, the swollen breast gradually shrinks and the pain is relieved. After taking it continuously for 6 months, the current condition is getting better and better.

Embodiments of the present invention

The content of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the embodiments of the present invention are only used to illustrate the present invention but not to limit the present invention. Without departing from the technical ideas of the present invention, various substitutions and changes made based on common technical knowledge and conventional means in the field, All should be included in the scope of the present invention.

Example 2

A patient from Si’an Town, Changxing County, Zhejiang Province, Wu Mou, female, 41 years old, went to the County People’s Hospital for examination due to left breast swelling and pain during menstruation. The Chinese Medicine Museum purchased the oral treatment of Wu Gu insect powder, twice a day, 3 grams each time. When the menstruation came in the next month, the pain was relieved. After 70 days of continuous use, the ultrasound examination results showed that the nodules at 2 o'clock on the left breast were reduced to 3.5*4mm.

Example 3

Patient in Nanxun District, Huzhou City, Zhejiang Province, Chen, female, 39 years old, bilateral breast hyperplasia has been for several years, affecting the lymphatic vessels under the arm, bilateral accessory breasts, usually chest pain, more pain before menstruation, menstrual friend It is recommended that I bought the five-grain insect powder for oral treatment at the Chinese Medicine Museum, twice a day, 2.5 grams each time. After taking it for one month, the pain was significantly reduced, and the accessory milk disappeared after four months.

Example 4

A patient from Hongxingqiao Town, Changxing County, Zhejiang Province, Xia, female, 46 years old, self-examined several lumps of different sizes in both breasts, and felt intense pain before menstruation. After a friend’s recommendation, I went to the Chinese Medicine Museum and purchased the five-grain insect powder for treatment, 6 grams a day, divided into two doses. After two months of continuous use, the pain gradually disappeared. Four months later, no obvious nodules were found in the breasts after ultrasound examination. Lumps.

Example 5

A 35-year-old female patient from Huzhou City, Zhejiang Province, Chen, found during a physical examination in June 2018 that a 6*5mm hypoechoic echo was seen at 11 o'clock in the left breast and a 4*3mm hypoechoic echo was seen at 10 o'clock in the right breast. . On both sides of the axillary scan, several lymph node echoes can be seen, the larger 14mmx4mm on the left and the larger 13mmx4mm on the right. Take Wu Gu Chong twice a day, 2.5 grams each time. After 70 days of continuous use, the breast tenderness and pain will be significantly improved. After taking it for 6 months, the ultrasound examination again showed that there was a 3*2.6mm hypoechoic nodule at 11 o'clock in the left breast, and the right breast nodule disappeared, and there was no obvious swollen lymph node echo.

The present invention also establishes a mammary hyperplasia rat model through estradiol benzoate combined with progesterone, and takes the clinical medication Rupixiao tablet as a positive drug to observe the effect of the double extract of the alcoholic water extract of Cephalocarpus on the height and diameter of the nipple and the mammary gland tissue of the rat with hyperplasia of mammary glands. As well as the influence of hormone levels of estradiol and prolactin, it is used to guide the clinical medication of Wu Gu Chong.

1 Materials and methods

1.1 Experimental animals

65 SPF female SD (Sprage-Dawley) rats, non-pregnant, weighing 180～200g, were raised in the Experimental Animal Center of Zhejiang University of Traditional Chinese Medicine; the laboratory room temperature was controlled at (22±3) ℃, and the relative humidity was 45%～55 %, the animals drink freely, and start the experiment after 1 week of adaptive feeding.

1.2 Reagents and drugs

Estradiol benzoate injection (Ningbo Second Hormone Factory, batch number: 110252511); Progesterone injection (Ningbo Second Hormone Factory, batch number: 110251670); Rupixiao tablets (Liaoning Shangyao Good Nurse Pharmaceutical Group Co., Ltd. , Batch number: 190413); grain insects (batch number: 20191013); physiological saline (Hangzhou Minsheng Pharmaceutical Co., Ltd., batch number: A1901152)

1.3 Experimental grouping and establishment of rat HMG model

65 SD rats were randomly divided into 6 groups, namely the blank control group (group A), the HMG model group (group B), the positive drug Rupixiao tablet (group C), the Wuguchong high-dose group (group D), Wu Gu Chong medium dose group (E group), Wu Gu Chong low dose group (F group). The modeling method is as follows: rats in the blank group were injected intramuscularly with normal saline injection 0.25mL/(Kg·d) in the right lower limbs for a total of 30 days; SD rats in the non-blank group were injected with estradiol benzoate intramuscularly in the lower limbs at a dose of 0.5 mg/(Kg·d), after 25 days of continuous injection, each rat was changed to intramuscular injection of progesterone at a dose of 5.0 mg/(Kg·d) for a total of 5 days ^[11,12,13] . Thirty days after the establishment of the model, the rat's nipples were straight, firm, red and enlarged. Randomly select 5 rats in the non-blank group, take a second pair of breast tissue for tissue section, HE staining, and use histological "gold standard" to determine whether a rat model of mammary gland hyperplasia is established.

1.4 Group dosage and administration

After successful establishment of the rat HMG model, the rats were administered and treated according to the groups. The dosage and method are as follows: Group A and Group B were given distilled water, 1 mL/100g, once a day; Group C was given Rupixiao tablet powder by intragastric administration Aqueous solution prepared with distilled water, the dose is 960mg/(Kg·d), 1 mL/100g, once a day; Group D is the high-dose group of the high-dose alcohol-water double-extract of Cephalosporium distilled by gavage, 1620mg/Kg, 1 mL/100g, once a day, the medium dose group of E group, 810mg/Kg, 1 mL/100g, once a day, group F, low-dose group, 405mg/Kg, 1 mL/100g, once a day, continuous intragastric administration for 27 days.

1.5 Observation indicators and methods

1.5.1 Measurement of nipple diameter and height Use vernier calipers to accurately measure the diameter and height of the second pair of breast nipple in each period after the initial, modeling, and treatment of the rat and take pictures.

1.5.2 Determination of organ index After the last gastric gavage, the rats were sacrificed after 12 hours of fasting. The uterus of each rat was taken out, and the mass was precisely weighed with an analytical electronic balance to calculate the organ index.

Organ index = organ mass (g)/weight (g)

1.5.3 Determination of hormones After 12 hours of fasting after the last gavage, the rats were anesthetized with 3% pentobarbital, blood was collected from the heart, the blood was left for 1 hour, centrifuged at 3000 r/min for 15 minutes, and the serum was collected and aliquoted to 1.5 mL EP tube, stored at -20°C, is used to detect estradiol (E ₂ ) and prolactin (PRL) content. The determination method is carried out in accordance with the kit instructions.

1.6 Statistical methods

Use SPSS17.0 software to process the experimental data. Single factor comparisons among multiple groups were first used to analyze the normal distribution of the single factor, and then statistically performed by the LSD method. P <0.05 is considered statistically significant.

2. Results and analysis

2.1 The effect of Wu Gu Chong on the body weight of rats with hyperplasia of mammary glands

It can be seen from Table 1 and Figure 1 that there was no significant difference in the initial body weight of rats in each group ( P >0.05). The weight of rats in the normal group continued to increase during the experiment. After 30 days of modeling, compared with the normal control group, the weight of the other modeled female rats was significantly reduced ( P <0.01), indicating that estradiol benzoate and corpus luteum were injected Ketoestrogens inhibit the growth of SD female rats. After intragastric administration for 27 days, compared with the normal control group, the body weight of the female rats in the other groups was significantly reduced ( P <0.01). Compared with the model group, there was no significant difference in the body weight of the female rats in the other treatment groups ( P >0.1), indicating that the high, medium and low doses of the positive drug Rupixiao Tablets and Wu Gu Chong can not alleviate the growth inhibition of female rats caused by exogenous high levels of estrogen.

2.2 The effect of Wu Gu insect on the height and diameter of the rat's nipple

It can be seen from Table 2 and Figure 2 that there was no significant difference in the initial nipple height of rats in each group ( P >0.1). Thirty days after modeling, compared with the blank control group, the nipple height of the other groups increased significantly ( P <0.01), indicating that injection of estradiol benzoate and progesterone estrogen can promote the growth of SD female rats' nipple. After 27 days of intragastric administration, compared with the normal control group, the nipple height of the model group was significantly increased ( P <0.01), and the nipple height of the other administration groups was significantly decreased ( P <0.01); the positive drug group, Wu Gu Chong high, medium There was no significant difference between the low doses ( P >0.05). It shows that Wu Gu Chong can treat the increase of nipples caused by the increase of exogenous estrogen levels, and the efficacy is equivalent to that of positive drugs.

It can be seen from Table 3 and Figure 3 that there was no significant difference in the initial nipple diameter of rats in each group ( P >0.1). Thirty days after modeling, compared with the blank control group, the nipple height of the other groups increased significantly ( P <0.01), indicating that injection of estradiol benzoate and progesterone estrogen can promote the growth of SD female rats' nipple. After 27 days of intragastric administration, compared with the normal control group, the nipple diameter of the model group was significantly increased ( P <0.01), and the nipple height of the other administration groups was significantly decreased ( P <0.01); the positive drug group, Wu Gu Chong high, medium There was no significant difference between the low doses ( P >0.05). It shows that Wu Gu Chong can treat the phenomenon of nipple enlargement caused by the increase of exogenous estrogen level, and the efficacy is equivalent to that of the positive medicine.

2.3 The effect of Wu Gu Chong on the uterine index

Excessive estrogen will thicken the uterine wall. The results show that compared with the normal control group, the uterine index of the model group was significantly increased ( P <0.01), and compared with the model group, the uterine index of each administration group was significantly decreased ( P <0.01); there was no significant difference between the positive drug group and the high, medium and low doses of Wu Gu Chong ( P > 0.05). It shows that Wu Gu Chong can treat the thickening of the uterus caused by the increase of exogenous estrogen levels, and the efficacy is equivalent to that of the positive medicine.

2.4 The effect of Wu Gu Chong on serum estradiol (E2) and prolactin (PRL) levels in rats

From Table 5 and Figure 5, the analysis of estradiol levels in rats shows that compared with the normal control group, the estradiol level of the model group was significantly increased ( P <0.01). Compared with the model group, the estradiol level of the other administration groups The level of diol was significantly reduced ( P <0.01). Compared with the positive drug group, the medium-dose group of Wu Gu Chong was significantly increased (P <0.05), and there was no significant difference between the high-dose and low-dose Wu Gu Chong ( P > 0.05). The results show that the high-dose and low-dose groups of Wu Gu Chong can significantly reduce the level of estradiol hormone in rats with hyperplasia of mammary glands, and the efficacy is equivalent to that of the positive drugs.

Analysis of the results of rat prolactin levels showed that compared with the normal control group, the prolactin level of the model group was significantly increased ( P <0.01), and compared with the model group, the prolactin levels of the other administration groups were significantly reduced ( P < 0.01). Compared with the positive drug group, there was no significant difference in the high, medium and low doses of Wu Gu Chong ( P > 0.1). The results showed that the high-dose, medium-, and low-dose of Wu Gu Chong can significantly reduce the level of prolactin hormone in rats with hyperplasia of mammary glands, and the efficacy is equivalent to that of the positive drugs.

The present invention establishes a rat model of mammary gland hyperplasia through estradiol benzoate combined with progesterone. The clinical drug Rupixiao tablet is used as a positive drug to observe the effect of the alcohol-water double extract of arachnids on the nipple height and diameter, mammary gland tissue and The effect of estradiol and prolactin on hormone levels. This experiment is divided into 3 stages, namely, before modeling of breast hyperplasia, after modeling, and after intragastric administration.

Before modeling, the initial body weight of rats in each group had no significant difference ( P >0.05). The weight of rats in the normal group continued to increase during the modeling period. After 30 days of modeling, compared with the normal control group, the weight of the rest of the modeled female rats was significantly reduced ( P <0.01), indicating that the injection of estradiol benzoate and the Progesterone estrogen can inhibit the growth of SD female rats, and its modeling is more successful from the appearance observation. After intragastric administration for 27 days, compared with the normal control group, the body weight of the female rats in the other groups was significantly reduced ( P <0.01). Compared with the model group, there was no significant difference in the body weight of the female rats in the other treatment groups ( P >0.1), indicating that the high, medium and low doses of the positive drug Rupixiao Tablets and Wu Gu Chong can not alleviate the growth inhibition of female rats caused by exogenous high levels of estrogen.

Regarding the nipple height and diameter, the initial nipple height and diameter of rats in each group had no significant difference ( P >0.1). Thirty days after modeling, compared with the blank control group, the nipple height and diameter of the other groups were significantly increased ( P <0.01), indicating that injection of estradiol benzoate and progesterone estrogen can promote the growth of SD female rats' nipple. After 27 days of intragastric administration, compared with the normal control group, the nipple diameter of the model group was significantly increased ( P <0.01), and the nipple height of the other administration groups was significantly decreased ( P <0.01); the positive drug group, Wu Gu Chong high, medium There was no significant difference between the low doses ( P >0.05). It shows that Wu Gu Chong can treat the phenomenon of nipple enlargement caused by the increase of exogenous estrogen level.

Excessive estrogen will thicken the uterine wall. The results show that compared with the normal control group, the uterine index of the model group was significantly increased ( P <0.01), and compared with the model group, the uterine index of each administration group was significantly decreased ( P <0.01); there was no significant difference between the positive drug group and the high, medium and low doses of Wu Gu Chong ( P > 0.05). It shows that Wu Gu Chong can treat the thickening of the uterus caused by the increase of exogenous estrogen levels.

The hypothalamic-pituitary-gonadal axis is one of the components of the nerve structure and plays an important role in maintaining the stability of the internal environment and the normal physiological functions of the body ^[14] . Abnormal secretion of sex hormones can lead to many diseases, such as hyperprolactinemia. , Breast hyperplasia, uterine fibroids, etc. The levels of various sex hormones jointly regulate breast tissue, among which elevated levels of prolactin (PRL) are considered to be an important factor in causing breast hyperplasia ^[15,16] , and progesterone (PROG) levels promote acinar tissue and breast lobules , Estradiol (E ₂ ) increased levels inhibited the growth of fibrous tissue around ducts and mammary ducts. Sex hormone disorders often occur in breast hyperplasia models, which can regulate the balance of the hypothalamus-pituitary-gonadal axis to regulate the levels of E ₂ , PROG, and PRL, restore ovarian function, and eliminate lobular hyperplasia and acinar hyperplasia ^[5] . Analysis of the results of rat prolactin levels, we know that compared with the normal control group, the prolactin level of the model group was significantly increased ( P <0.01), compared with the model group, the prolactin levels of the other administration groups were significantly reduced ( P < 0.01). Compared with the positive drug group, there was no significant difference in the high, medium and low doses of Wu Gu Chong ( P > 0.1). It shows that the high dose, medium and low dose of Wu Gu Chong can significantly reduce the level of prolactin hormone in rats with hyperplasia of mammary glands. For estradiol, compared with the normal control group, the estradiol level of the model group was significantly increased ( P <0.01), and compared with the model group, the estradiol level of the other administration groups was significantly reduced ( P <0.01) ), compared with the positive drug group, the Wu Gu Chong medium dose group was significantly higher ( P <0.05), and there was no significant difference between the high and low doses of Wu Gu Chong ( P > 0.05). It shows that the high-dose and low-dose groups of Wu Gu Chong can significantly reduce the level of estradiol hormone in rats with hyperplasia of mammary glands.

In summary, the worm can regulate the disordered hormones in rats with mammary gland hyperplasia, and its main components have the function of balancing estrogen, and have protective and repairing effects on mammary gland hyperplasia.

Claims (10)

Application of Wu Gu Chong in the preparation of medicines for preventing and treating mammary gland hyperplasia. The application according to claim 1, wherein the grain worm is a grain worm extract. The application according to claim 3, characterized in that the five grain insect extract is an alcohol-water double extract of grain insect. Application of Wu Gu Chong in the treatment of nipple nodules caused by increased levels of exogenous estrogen. The application of Wu Gu Chong in the medicine for reducing the prolactin hormone level of mammary gland hyperplasia. The application according to claim 1, wherein the grain worm is a grain worm extract. The application according to claim 6, characterized in that the five grain insect extract is an alcohol water double extract of five grain insect. Application of Wu Gu Chong in the treatment of uterine thickening drugs caused by increased levels of exogenous estrogen. A medicine for preventing and treating hyperplasia of mammary glands, which contains grain worm or grain worm alcohol-water double extract. The medicine according to claim 9, characterized in that it also contains pharmaceutically acceptable conventional excipients.