Application of aspirin as the sole active ingredient in the preparation of drugs for treating endometrial hyperplasia

Abstract

The present invention discloses a new use of aspirin, specifically its use in the preparation of a drug for treating endometrial hyperplasia. In a model of estrogen-induced endometrial hyperplasia, aspirin effectively inhibits endometrial thickness in cynomolgus monkeys and mice. Aspirin significantly improves the symptoms of endometrial hyperplasia in both mice and cynomolgus monkeys, thus providing a new clinical application for aspirin.

Description

Technical Field

[0001] The present invention relates to a new application of aspirin, and in particular to an application of aspirin in preparing a medicine for treating endometrial hyperplasia. Background Art

[0002] Endometrial hyperplasia (EH) refers to a proliferative lesion characterized by proliferation of endometrial glands and / or an increase in the proportion of glandular stroma. It is a common gynecological disease with a potential for cancer. The etiology is generally believed to be related to long-term estrogen stimulation without progesterone antagonism. Clinically, EH is most common in perimenopausal women, and its incidence has increased significantly in young women of childbearing age in recent years. The main clinical manifestations of EH are abnormal uterine bleeding, such as prolonged menstruation, menorrhagia, intermenstrual bleeding, and postmenopausal bleeding. Prolonged and heavy bleeding can be complicated by anemia and infection, and infertility can occur in women of childbearing age. Some patients may also develop malignant transformation. 2%-10% of abnormal uterine bleeding and 3%-10% of postmenopausal bleeding are caused by EH. Because atypical EH has a potential for cancer and a high risk of becoming malignant, understanding its pathogenesis, early diagnosis, and appropriate treatment are crucial to preventing further progression.

[0003] Aspirin, also known as acetylsalicylic acid, was invented in 1897 as a derivative of salicylic acid. It is commonly used as an analgesic, antipyretic, and anti-inflammatory drug. However, its unique pharmacological effects in treating endometrial hyperplasia have not been reported to date. The present invention primarily discovered that aspirin significantly improves endometrial hyperplasia models in cynomolgus monkeys and mice.

[0004] The structural formula of aspirin is as follows:

[0005] Summary of the Invention

[0006] In order to address the drawbacks of existing clinical hormone therapy for the treatment of endometrial hyperplasia, such as poor compliance, contraindications, easy recurrence and breast diseases, the present invention proposes that aspirin can be used to treat endometrial hyperplasia-related diseases.

[0007] Intraperitoneal administration of aspirin significantly alleviated endometrial hyperplasia in cynomolgus monkeys and mice induced by subcutaneous estrogen injection. The aspirin concentrations used were 3.2 mg / kg and 10 mg / kg, respectively. These results suggest that aspirin can improve endometrial hyperplasia in both monkeys and mice, and therefore may be used as a therapeutic agent for endometrial hyperplasia.

[0008] Beneficial effects

[0009] The present invention is the first to discover that aspirin can treat endometrial hyperplasia, which is a new indication for aspirin, as follows:

[0010] Endometrial hyperplasia (EH) refers to a proliferative lesion characterized by proliferation of endometrial glands and / or an increase in the proportion of glandular stroma. It is a common gynecological disease with a potential for malignant transformation. The etiology is generally believed to be related to long-term estrogen stimulation without progestogen antagonism. The main clinical manifestations of EH are abnormal uterine bleeding, such as prolonged menstruation, menorrhagia, intermenstrual bleeding, and postmenopausal bleeding. Prolonged and heavy bleeding can be complicated by anemia and infection, infertility in women of childbearing age, and malignant transformation in some patients. Currently, multiple treatment options are available for EH, each tailored to the extent of EH, patient age, and fertility needs. For patients without atypical EH, medications such as progestogens can be considered. However, long-term use is associated with poor patient compliance, contraindications, high recurrence rates, and the risk of breast disease. EH should not be used in patients with breast cancer and requires repeated diagnostic curettage or hysteroscopy, which can damage the endometrium and compromise fertility. Patients with atypical endometrial hyperplasia require endometrial ablation or even hysterectomy, which is even less acceptable to patients. Therefore, finding new intervention targets and therapeutic drugs for endometrial hyperplasia is a common gynecological disease that needs to be solved urgently.

[0011] The present invention uses aspirin to treat estrogen-induced endometrial hyperplasia models in mice and cynomolgus monkeys. Experiments show that intraperitoneal injection of aspirin can significantly reduce the weight of uterine tissue and the proportion of endometrial glandular stroma in mice and cynomolgus monkeys.

[0012] The above results suggest that aspirin can significantly improve the symptoms of endometrial hyperplasia in mice and crab-eating monkeys, so it can be used to treat endometrial hyperplasia. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 Aspirin inhibits estrogen-induced endometrial hyperplasia in mice.

[0014] C57BL / 6j female mice, 8-10 weeks old, were housed in an SPF animal room at 21±2°C, with free access to water and food, and a 12-hour day and night cycle. The mice were randomly divided into 4 groups: a normal group, an estradiol (50 μg / kg) group, an aspirin (10 mg / kg) group, and a progesterone group (10 mg / kg). The estradiol group, the progesterone group, and the aspirin group received a subcutaneous injection of 100 μl of estradiol every day. In addition, the aspirin group received an intraperitoneal injection of aspirin, and the progesterone group received an intraperitoneal injection of progesterone every day. Once a day, 100 μl each time, and the normal group was given an equal amount of solvent. After 21 days of treatment, the experimental animals were euthanized and their uteri were taken for photographing.

[0015] Figure 2 Aspirin inhibits estrogen-induced increase in uterine weight in mice.

[0016] Mice were randomly divided into four groups: a normal group, an estradiol (50 μg / kg) group, an aspirin (10 mg / kg) group, and a progestin (10 mg / kg) group. The estradiol, progestin, and aspirin groups received daily subcutaneous injections of 100 μl of estradiol. The aspirin group also received intraperitoneal injections of aspirin, while the progestin group received daily intraperitoneal injections of progestin. After daily administration for 21 days, the animals were euthanized and their uteri were weighed. Results are expressed as mean ± standard deviation. **: P < 0.01, ***: P < 0.001 vs. estradiol (Student's t test).

[0017] Figure 3 Results of hematoxylin-eosin staining (HE staining) of mouse endometrial tissue sections.

[0018] Mice were randomly divided into four groups: a normal control group, an estradiol (50 μg / kg) group, an aspirin (10 mg / kg) group, and a progestin (10 mg / kg) group. The estradiol, progestin, and aspirin groups received daily subcutaneous injections of 100 μl of estradiol. The aspirin group also received intraperitoneal injections of aspirin; the progestin group received daily intraperitoneal injections of progestin. After 21 days of daily dosing, the animals were euthanized, and their uteri were fixed, dehydrated, embedded, and sectioned for HE staining.

[0019] Figure 4 Aspirin inhibits the estrogen-induced increase in the glandular-stromal ratio of the mouse endometrium.

[0020] Mice were randomly divided into four groups: a normal group, an estradiol (50 μg / kg) group, an aspirin (10 mg / kg) group, and a progestin (10 mg / kg) group. The estradiol, progestin, and aspirin groups received daily subcutaneous injections of 100 μl of estradiol. The aspirin group also received intraperitoneal aspirin injections; the progestin group received daily intraperitoneal progestin injections. After 21 days of daily dosing, the animals were euthanized, and their uteri were fixed, dehydrated, embedded, and sectioned for HE staining. The glandular-stromal ratio of endometrial tissue was observed and analyzed. Results are expressed as mean ± standard deviation. **: P < 0.01, ***: P < 0.001 vs. estradiol (Student's t test).

[0021] Figure 5 Aspirin inhibits estrogen-induced endometrial hyperplasia in cynomolgus monkeys.

[0022] Cynomolgus monkeys were randomly divided into three groups: a control group, an estrogen group, and an aspirin group. The estrogen and aspirin groups received a daily subcutaneous injection of 500 μL of estrogen (64 μg / kg). Additionally, the aspirin group received an intraperitoneal injection of 4 mL of aspirin (3.2 mg / kg) once daily. The control group received an equal volume of solvent. Treatment lasted for 5 weeks, and the animals underwent weekly ultrasound examinations.

[0023] Figure 6 HE staining results of endometrial tissue sections of cynomolgus monkeys.

[0024] Cynomolgus monkeys were randomly divided into three groups: a control group, an estrogen group, and an aspirin group. The estrogen and aspirin groups received a daily subcutaneous injection of 500 μL of estrogen (64 μg / kg). Additionally, the aspirin group received an intraperitoneal injection of 4 mL of aspirin (3.2 mg / kg) once daily. The control group received an equal volume of solvent. Treatment lasted for 5 weeks. Finally, the animals were euthanized, and their uteri were fixed, dehydrated, embedded, and sectioned for hematoxylin and eosin staining.

[0025] Figure 7 Aspirin inhibits estrogen-induced increase in endometrial glandular-stromal ratio in cynomolgus monkeys.

[0026] Cynomolgus macaques were randomly divided into three groups: a control group, an estrogen group, and an aspirin group. The estrogen and aspirin groups received a daily subcutaneous injection of 500 μL of estrogen (64 μg / kg). In addition, the aspirin group received an intraperitoneal injection of 4 mL of aspirin (3.2 mg / kg) once daily. The control group received an equal amount of solvent. After five weeks of treatment, the animals were euthanized, and their uteri were fixed, dehydrated, embedded, and sectioned for HE staining. The glandular stroma ratio of the endometrial tissue was observed and calculated. The results are expressed as mean ± standard deviation. **: P < 0.01, ***: P < 0.001 vs. Estradiol (Student's t test). DETAILED DESCRIPTION

[0027] Example 1: Aspirin inhibits endometrial hyperplasia induced by subcutaneous estrogen injection in mice

[0028] Technical methods: Mouse endometrial hyperplasia model induced by subcutaneous estrogen injection, HE staining

[0029] Mice were randomly divided into four groups: a normal group, an estradiol (50 μg / kg) group, an aspirin (10 mg / kg) group, and a progestin (10 mg / kg) group. The estradiol, progestin, and aspirin groups received a daily subcutaneous injection of 100 μl of estradiol. The aspirin group also received an intraperitoneal injection of aspirin. The progestin group received a daily intraperitoneal injection of progestin. The injections were administered once daily, 100 μl each time. The control group received an equal amount of solvent. After 21 days of treatment, the animals were euthanized, and their uteri were removed for photographing and weighing. The sections were fixed, dehydrated, embedded, and stained with HE to observe endometrial tissue proliferation.

[0030] Example 2: Aspirin inhibits endometrial hyperplasia induced by subcutaneous estrogen injection in cynomolgus monkeys

[0031] Technical methods: Mouse endometrial hyperplasia model induced by subcutaneous estrogen injection, HE staining

[0032] Four-year-old crab-eating macaques were housed in an SPF animal room at 21±2°C with free access to water and food, and a 12-hour day and night cycle. The crab-eating macaques were randomly divided into three groups: a control group, an estrogen (64μg / kg) group, and an aspirin (3.2mg / kg) group. The estrogen group and the aspirin group received a subcutaneous injection of 500μL of estrogen daily. In addition, the aspirin group received an intraperitoneal injection of aspirin once a day, 4mL each time, and the control group received an equal amount of solvent. The animals were treated for 5 weeks and underwent ultrasound examinations every week. Finally, the animals were euthanized, and their uteri were fixed, dehydrated, embedded, and sectioned for HE staining to observe the proliferation of their endometrial tissue. Analysis of aspirin pharmacological experiment results:

[0033] Aspirin improves estrogen-induced endometrial hyperplasia in mice.

[0034] like Figure 1 As shown, after 21 days, the uterine tissue of mice in the estrogen group was enlarged compared to the control group. The enlargement of uterine tissue in the positive control drug progesterone group was improved, and the enlargement of uterine tissue in the mice in the aspirin group was significantly improved. Aspirin can significantly improve estrogen-induced uterine enlargement in mice.

[0035] like Figure 2 As shown in the figure, the uterine weights of mice in each group were statistically analyzed. Compared with the control group, the uterine weights of mice in the estrogen group were significantly increased. The positive control drugs progesterone and aspirin also significantly improved the uterine weights of mice. Therefore, aspirin can significantly improve estrogen-induced uterine enlargement in mice.

[0036] like Figure 3As shown, HE staining results showed that compared with the control group, the endometrial glands in the estrogen group were overgrown, while the endometrial glands in the progesterone and aspirin groups were improved. Aspirin can significantly improve the estrogen-induced endometrial gland growth in mice.

[0037] like Figure 4 As shown in the figure, the ratio of endometrial glands to stroma in mice was statistically analyzed based on the HE staining results. The results showed that compared with the control group, the ratio of endometrial glands to stroma in the estrogen group was significantly increased. The positive control drugs progesterone and aspirin both significantly reduced the ratio of endometrial glands to stroma in mice. Therefore, aspirin can significantly improve estrogen-induced endometrial gland growth in mice.

[0038] like Figure 5 As shown, after 5 weeks of treatment and weekly ultrasound examinations of the experimental animals, the endometrial thickness of the estrogen-treated cynomolgus monkeys increased compared to the control group, and the endometrial hyperplasia of the cynomolgus monkeys in the aspirin-treated group was improved. Aspirin can significantly improve estrogen-induced endometrial hyperplasia in cynomolgus monkeys.

[0039] like Figure 6 As shown, HE staining results showed that compared with the control group, the endometrial glands in the estrogen group of cynomolgus monkeys were overgrown, while the endometrial gland growth in the aspirin group was improved. Aspirin can significantly improve estrogen-induced endometrial gland growth in cynomolgus monkeys.

[0040] like Figure 7 As shown in the figure, HE staining results showed that the ratio of endometrial glandular stroma in cynomolgus monkeys was significantly increased in the estrogen group compared with the control group, while aspirin significantly reversed the increase in the ratio of endometrial glandular stroma in cynomolgus monkeys. Therefore, aspirin can significantly improve estrogen-induced endometrial glandular growth in cynomolgus monkeys.

Claims

1. Use of aspirin as the sole active ingredient in the preparation of a drug for treating endometrial hyperplasia, wherein the structural formula of aspirin is as follows:

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