阅读说明：本技术 阿司匹林在制备治疗子宫内膜增生药物中的应用 (Application of aspirin in preparation of medicine for treating endometrial hyperplasia ) 是由 孙洋 徐强 瞿娇 丁衍 张涛 于 2021-08-20 设计创作，主要内容包括：本发明公开了阿司匹林的新用途,具体涉及阿司匹林在制备治疗子宫内膜增生药物中的应用。针对雌激素诱导的子宫内膜增生的疾病模型,阿司匹林能有效抑制食蟹猴以及小鼠的子宫内膜厚度。阿司匹林能显著改善小鼠以及食蟹猴子宫内膜增生的症状,故其可应用于治疗子宫内膜增生,临床上增加了阿司匹林的新用途。(The invention discloses a new application of aspirin, and particularly relates to an application of aspirin in preparation of a medicine for treating endometrial hyperplasia. Aiming at disease models of estrogen-induced endometrial hyperplasia, aspirin can effectively inhibit the thickness of the endometrium of cynomolgus monkeys and mice. Aspirin can remarkably improve the endometrial hyperplasia symptoms of mice and cynomolgus monkeys, so that aspirin can be applied to treating endometrial hyperplasia, and the new application of aspirin is increased clinically.)

1. The application of aspirin in preparing medicine for treating endometrial hyperplasia is disclosed, wherein the structural formula of aspirin is as follows:

One, the technical field

The invention relates to a new application of aspirin, in particular to an application of aspirin in preparation of a medicine for treating endometrial hyperplasia.

Second, background Art

Endometrial hyperplasia (endometrial hyperplasia) refers to a proliferative lesion of the endometrium with an increased proportion of hyperplasia of the endometrium and/or interstitial gland, is a common gynecological disease with a certain tendency to canceration, and the pathogenic factor is generally considered to be related to estrogen stimulation without long-term progestational hormone antagonism. Clinical endometrial hyperplasia is mostly seen in perimenopausal women, and the incidence rate of young women in reproductive age is also obviously increased in recent years. Endometrial hyperplasia is characterized clinically by abnormal uterine bleeding (abnormal uterine bleeding), such as menostaxis, menorrhagia, intermenstrual bleeding and postmenopausal bleeding, with long bleeding time and large amount of bleeding possibly complicated by anemia and infection, infertility of patients in childbearing age, and cachexia of some patients. 2% -10% of abnormal uterine bleeding and 3% -10% of postmenopausal bleeding are caused by endometrial hyperplasia. Because atypical hyperplasia in endometrial hyperplasia has a canceration trend and risk, understand its pathogenesis, carry on early diagnosis and reasonable treatment to it, it is important to prevent and treat the further progress of pathological change.

Aspirin, also known as acetylsalicylic acid, was invented in 1897 as a derivative of salicylic acid, a compound commonly used as an analgesic, antipyretic and anti-inflammatory agent. The unique pharmacological action of the compound for treating the endometrial hyperplasia is not reported so far. The invention mainly discovers that aspirin has a remarkable improvement effect on endometrial hyperplasia models of cynomolgus monkeys and mice.

The structural formula of aspirin is as follows:

third, the invention

In order to solve the defects of the existing hormone therapy for clinically treating the endometrial hyperplasia, such as poor compliance, medication contraindication, easy relapse, breast disease and the like, the invention provides the case that aspirin can be used for treating the diseases related to the endometrial hyperplasia.

For estrogen subcutaneous injection induced endometrium hyperplasia models of cynomolgus monkeys and mice, the administration of aspirin in abdominal cavity can obviously relieve the endometrium hyperplasia phenomenon of the cynomolgus monkeys and the mice, and the concentration of the aspirin injection is 3.2mg/kg and 10mg/kg respectively. The results suggest that aspirin can improve endometrial hyperplasia in cynomolgus monkeys and mice, and thus it can be applied to a therapeutic agent for endometrial hyperplasia.

Fourthly, beneficial effects

The invention discovers for the first time that aspirin can treat endometrial hyperplasia, and is a new adaptive disease of aspirin, which is as follows:

the endometrial hyperplasia refers to endometrial hyperplasia and/or endometrial proliferative lesion with increased glandular-interstitial proportion, is a common gynecological disease with a certain canceration tendency, and usually pathogenic factors are considered to be related to estrogen stimulation without progestational hormone antagonism for a long time. Endometrial hyperplasia is characterized clinically by abnormal uterine bleeding, such as prolonged menstrual period, menorrhagia, intermenstrual bleeding and postmenopausal bleeding, which may be complicated by anemia and infection due to long bleeding time and large amount, infertility of patients in childbearing age, and degeneration of some patients. At present, various methods for treating endometrial hyperplasia exist, and a personalized scheme needs to be established according to the endometrial hyperplasia degree, the age of a patient and the fertility requirement. For the patients without atypical hyperplasia, the application of progestogen and other medicaments can be considered, but the patients with long-term administration have the disadvantages of poor compliance, medication contraindication, easy relapse, breast diseases and the like, and the medicine cannot be used for the patients with breast malignant tumors, repeated diagnostic uterine curettage or hysteroscopy is needed, and endometrium can be damaged to influence the fertility. For the patient with atypical hyperplasia of the endometrium, an endometrium removal operation is required, even the uterus is cut off, and the patient is not easy to accept the method. Therefore, the search for new intervention targets and therapeutic drugs for endometrial hyperplasia is a problem to be solved urgently for common gynecological diseases at present.

The invention adopts aspirin to treat estrogen-induced endometrial hyperplasia models of mice and cynomolgus monkeys, and experiments show that the weight of uterine tissues of the mice and the cynomolgus monkeys and the proportion of endometrial gland interstitial substances can be obviously reduced by injecting the aspirin into abdominal cavities.

The results suggest that aspirin can significantly improve the symptoms of endometrial hyperplasia in mice and cynomolgus monkeys, so that it can be used for treating endometrial hyperplasia.

Description of the figures

Figure 1 aspirin inhibits estrogen-induced endometrial hyperplasia in mice.

C57BL/6j female mice, 8-10 weeks old, were bred in SPF-class animal houses at 21 + -2 deg.C with free drinking water and feeding, alternating day and night for 12 h. The mice were randomly divided into 4 groups, a normal group, an estradiol (50. mu.g/kg) group, an aspirin (10mg/kg) group, and a progestin group (10 mg/kg). Estradiol group, progestogen group and aspirin group, 100 μ l of estradiol was subcutaneously injected daily, in addition, aspirin was intraperitoneally injected in the aspirin group, and progestogen was intraperitoneally injected daily in the progestogen group. The normal group was given an equal amount of solvent once a day, 100. mu.l each time. 21 days after the administration treatment, the experimental animals were euthanized and the uterus was photographed

Figure 2 aspirin inhibits estrogen-induced increase in uterine weight in mice.

The mice were randomly divided into 4 groups, a normal group, an estradiol (50. mu.g/kg) group, an aspirin (10mg/kg) group, and a progestin group (10 mg/kg). Estradiol group, progestogen group and aspirin group, 100 μ l of estradiol was subcutaneously injected daily, in addition, aspirin was intraperitoneally injected in the aspirin group, and progestogen was intraperitoneally injected daily in the progestogen group. Daily dosing was continued for 21 days, after which the experimental animals were euthanized and their uteri were weighed. Results are expressed as mean ± standard deviation. **: p <0.01, x: p <0.001vs Estradiol; (Student's-ttest).

FIG. 3 results of hematoxylin-eosin staining (HE staining) of mouse endometrial tissue sections.

The mice were randomly divided into 4 groups, a normal group, an estradiol (50. mu.g/kg) group, an aspirin (10mg/kg) group, and a progestin group (10 mg/kg). Estradiol group, progestogen group and aspirin group, 100 μ l of estradiol is subcutaneously injected every day, and in addition, aspirin group is required to be intraperitoneally injected; progestogen group daily intraperitoneal injections of progestogen were given. Daily dosing, after 21 days, experimental animals were euthanized and uterus-fixed, dehydrated, embedded sections were taken and HE-stained.

Figure 4 aspirin inhibits estrogen-induced increase in the endometrial glandular interstitial proportion in mice.

The mice were randomly divided into 4 groups, a normal group, an estradiol (50. mu.g/kg) group, an aspirin (10mg/kg) group, and a progestin group (10 mg/kg). Estradiol group, progestogen group and aspirin group, 100 μ l of estradiol is subcutaneously injected every day, and in addition, aspirin group is required to be intraperitoneally injected; progestogen group daily intraperitoneal injections of progestogen were given. Daily administration is carried out, after 21 days, the experimental animals are euthanized, fixed dehydrated embedding sections of the uterus are taken, HE staining is carried out, and the endometrial tissue glandular stroma proportion is observed and counted. Results are expressed as mean ± standard deviation. **: p <0.01, x: p <0.001vs Estradiol; (Student's-t test).

Figure 5 aspirin inhibits estrogen-induced endometrial hyperplasia in cynomolgus monkeys.

Cynomolgus monkeys were randomly divided into 3 groups, a control group, an estrogen group, and an aspirin group. The estrogen group and aspirin group were subcutaneously injected with 500. mu.L of estrogen (64. mu.g/kg) per day, and in addition, the aspirin group was intraperitoneally injected with 4mL of aspirin (3.2mg/kg) once per day, and the control group was administered with the same amount of solvent. Treatment was administered for 5 weeks and the experimental animals were examined ultrasonically weekly.

FIG. 6 results of HE staining of endometrial tissue sections from cynomolgus monkeys.

Cynomolgus monkeys were randomly divided into 3 groups, a control group, an estrogen group, and an aspirin group. The estrogen group and aspirin group were subcutaneously injected with 500. mu.L of estrogen (64. mu.g/kg) per day, and in addition, the aspirin group was intraperitoneally injected with 4mL of aspirin (3.2mg/kg) once per day, and the control group was administered with the same amount of solvent. The treatment is given for 5 weeks, and finally, the experimental animals are euthanized, and the uterus is taken for fixed dehydration embedding section and HE staining is carried out.

Figure 7 aspirin inhibits estrogen-induced increase in endometrial glandular interstitial proportion in cynomolgus monkeys.

Cynomolgus monkeys were randomly divided into 3 groups, a control group, an estrogen group, and an aspirin group. The estrogen group and aspirin group were subcutaneously injected with 500. mu.L of estrogen (64. mu.g/kg) per day, and in addition, the aspirin group was intraperitoneally injected with 4mL of aspirin (3.2mg/kg) once per day, and the control group was administered with the same amount of solvent. The treatment is given for 5 weeks, and finally, the experimental animals are euthanized, and the uterus is taken for fixed dehydration embedding section and HE staining is carried out. Observing and counting the endometrial tissue glandular interstitial proportion. Results are expressed as mean ± standard deviation. **: p <0.01, x: p <0.001vs Estradiol; (Student's-t test).

Sixth, detailed description of the invention

Example 1: aspirin inhibits estrogen subcutaneous injection induced endometrial hyperplasia in mice

The technical method comprises the following steps: mouse endometrial hyperplasia model induced by estrogen subcutaneous injection and HE staining

The mice were randomly divided into 4 groups, a normal group, an estradiol (50. mu.g/kg) group, an aspirin (10mg/kg) group, and a progestin group (10 mg/kg). Estradiol group, progestogen group and aspirin group, 100 μ l of estradiol is subcutaneously injected every day, and in addition, aspirin group is required to be intraperitoneally injected; progestogen group daily intraperitoneal injections of progestogen were given. The control group was given an equal amount of solvent once a day, 100. mu.l each time. After 21 days of administration treatment, the experimental animals are euthanized, the uterus of the experimental animals is taken for photographing and weighing, the dehydrated embedded section is fixed, and HE staining is carried out to observe the proliferation condition of the endometrial tissue of the experimental animals.

Example 2: aspirin inhibits estrogen subcutaneous injection induced endometrial hyperplasia of cynomolgus monkey

The technical method comprises the following steps: mouse endometrial hyperplasia model induced by estrogen subcutaneous injection and HE staining

The cynomolgus monkey is taken and aged 4 years old, and is bred in an SPF animal house at the temperature of 21 +/-2 ℃, and the cynomolgus monkey is taken by freely drinking water and is alternated day and night for 12 hours. Cynomolgus monkeys were randomized into 3 groups, control group, estrogen (64 μ g/kg) group, aspirin (3.2mg/kg) group. The estrogen group and aspirin group were injected subcutaneously with 500. mu.L of estrogen per day, and in addition, the aspirin group was injected intraperitoneally with 4mL of aspirin once per day, and the control group was given the same amount of solvent. The administration treatment is carried out for 5 weeks, the experimental animals are subjected to ultrasonic examination every week, finally, the experimental animals are euthanized, the uterus of the experimental animals is taken for fixed dehydration embedding section, and HE staining is carried out to observe the proliferation condition of the endometrial tissues of the experimental animals. Seventhly, analyzing pharmacological experiment results of aspirin:

the improvement effect of aspirin on estrogen-induced endometrial hyperplasia models in mice.

As shown in fig. 1, after 21 days, compared with the control group, the uterine tissue of the mice with estrogen group was increased, the uterine tissue of the mice with positive control drug, progestogen group was increased, and the uterine tissue of the mice with aspirin group was significantly improved. Aspirin can obviously improve the mouse uterus enlargement induced by estrogen.

As shown in fig. 2, when the weights of the uteri of the mice in each group are counted, the weight of the uterus of the mice in the estrogen group is obviously increased compared with that of the control group, and the weight of the uterus of the mice is obviously improved by the progestogen and the aspirin which are positive control medicaments. Therefore, aspirin can significantly improve estrogen-induced uterine enlargement in mice.

As shown in fig. 3, HE staining results showed that endometrial gland overgrowth was improved in the estrogen group mice, and endometrial gland growth was improved in the progestogen group and aspirin group mice, compared to the control group. Aspirin can obviously improve the growth of mouse endometrium gland induced by estrogen.

As shown in figure 4, statistics is carried out on the ratio of endometrial glands to stroma of the mice according to the HE staining result, and the result shows that compared with the control group, the endometrial gland stroma ratio of the mice in the estrogen group is obviously increased, and the endometrial gland stroma ratio of the mice is obviously reduced by the progestogen and the aspirin which are positive control drugs. Therefore, aspirin can significantly improve estrogen-induced endometrial gland growth in mice.

As shown in fig. 5, when the treatment was performed for 5 weeks and the ultrasonic examination was performed on the experimental animals every week, the endometrial thickness of the cynomolgus monkey in the estrogen group was increased and the endometrial hyperplasia of the cynomolgus monkey in the aspirin-administered group was improved, as compared with the control group. The aspirin can obviously improve estrogen-induced endometrial hyperplasia of the cynomolgus monkey.

As shown in fig. 6, HE staining results showed that the endometrial glands of the cynomolgus monkey in the estrogen group were overgrown and the endometrial glands of the cynomolgus monkey in the aspirin group were improved compared with the control group. Aspirin can obviously improve the growth of endometrial glands of the cynomolgus monkey induced by estrogen.

As shown in fig. 7, according to HE staining results, statistics on the cynomolgus monkey endometrial gland interstitial proportion showed that the cynomolgus monkey endometrial gland interstitial proportion was significantly increased in the estrogen group compared to the control group, while aspirin significantly reversed the increase in the cynomolgus monkey endometrial gland interstitial proportion. Therefore, aspirin can significantly improve estrogen-induced endometrial gland growth in cynomolgus monkeys.