Application of dapam nitrile in prevention and / or treatment of radioactive intestinal injury

Abstract

The invention relates to the field of biology and medicine, and discloses application of dapam nitrile in prevention and / or treatment of radioactive intestinal injury. The inventor of the invention finds that dapam nitrile can protect radioactive intestinal injury induced by ionizing radiation local irradiation, including improvement of tissue pathological injury, promotion of intestinal crypt cell regeneration, inhibition of intestinal cell apoptosis, improvement of intestinal cell pyroptosis and reduction of serum inflammatory factor level; therefore, the compound can be used as a potential medicine for preventing and / or treating radioactive intestinal injury.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to the application of dapoxetine in preventing and / or treating radiation-induced intestinal injury. Background Art

[0002] Radiation-induced bowel injury is an intestinal complication caused by radiotherapy for malignant tumors in the pelvis, abdomen and other parts of the body. The lesions can affect the small intestine, rectum and colon. The main symptoms are abdominal pain, diarrhea, blood in the stool and other symptoms. In severe cases, there may be complications such as intestinal ulcers and even intestinal perforation. Acute radiation-induced bowel injury is a key factor in death and treatment failure of systemic radiation injury, and is a bottleneck restricting the successful treatment of extremely severe radiation sickness. Currently, about 50% of cancer survivors receive radiotherapy for abdominal and pelvic cancers, and the global burden of radiation-induced bowel injury will increase significantly in the future. Although a large number of studies have been conducted on the prevention and treatment of acute radiation-induced bowel injury, many studies on drug therapy, biological therapy, cell therapy, etc. are still in the preclinical stage, and there is currently no specific therapeutic drug for the treatment or prevention of acute radiation-induced bowel injury. Therefore, it is of great significance to study new strategies and drugs for the prevention and treatment of acute radiation-induced bowel injury.

[0003] Dapansutrile (OLT1177) is a β-sulfonyl nitrile compound that has been shown to treat gout attacks and experimental allergic asthma in mice. However, the role of OLT1177 in acute radiation-induced intestinal injury is still unclear. Summary of the invention

[0004] In order to overcome the above problems in the prior art, the present invention provides the use of dapanoxacil in preventing and / or treating radiation-induced intestinal injury.

[0005] The first aspect of the present invention provides the use of dapanoxacil in the preparation of a medicament for preventing and / or treating radiation-induced intestinal injury.

[0006] Dapanoxonitrile is 3-methylsulfonylpropionitrile, and its structure is shown below:

[0007]

[0008] In the present invention, the radiation intestinal injury refers to intestinal tissue damage caused by high-dose radiation exposure, which usually occurs within a few days after high-dose radiation exposure. The main characteristics include nausea, vomiting, abdominal pain, diarrhea, gastrointestinal bleeding, bacterial infection, and other acute gastrointestinal (GI) syndrome symptoms caused by local irradiation of ionizing radiation. The radiation intestinal injury is preferably intestinal injury caused by local irradiation of ionizing radiation. The radiation intestinal injury is a complication caused by radiotherapy of malignant tumors in the abdomen and / or pelvis, that is, the object of prevention and / or treatment is a patient with a malignant tumor. Generally, malignant tumors that require radiotherapy are ovarian malignant tumors, endometrial cancer, cervical cancer, pelvic malignant lymphoma and other types. The ionizing radiation energy absorption dose that is prone to radiation ovarian malignant tumors, endometrial cancer, cervical cancer, and pelvic malignant lymphoma damage is 12-16Gy / time (time is 1).

[0009] The second aspect of the present invention provides the use of dapanoxacil in the preparation of a drug for improving pathological damage of intestinal tissue caused by radiation (radiation intestinal damage).

[0010] The third aspect of the present invention provides the use of dapanoxacil in the preparation of a drug for promoting the regeneration of intestinal crypt cells (in radiation-induced intestinal injury).

[0011] The fourth aspect of the present invention provides the use of dapanoxacil in the preparation of a drug for inhibiting (radiation-induced) intestinal cell apoptosis.

[0012] The fifth aspect of the present invention provides the use of dapanoxacil in the preparation of a drug for inhibiting (radiation-induced) intestinal cell pyroptosis.

[0013] The sixth aspect of the present invention provides the use of dapanoxacil in the preparation of a drug for reducing the secretion of serum inflammatory factors (especially the secretion of serum inflammatory factors in patients with radiation-induced intestinal injury).

[0014] In a preferred embodiment of the present invention, the inflammatory factor is selected from at least one of TNF-α, IL-6, MCP-1, IL-10 and IFN-γ.

[0015] The "prevention and / or treatment" in the present invention includes: relief of symptoms related to intestinal damage, including: relief of inflammation and edema symptoms.

[0016] In the present invention, there is no particular requirement for the administration method of dapamoxil, but it is preferably at least one of intravenous injection, intramuscular injection and subcutaneous injection. Those skilled in the art can prepare dapamoxil into a specific dosage form, such as an injection, according to the administration method.

[0017] In the present invention, the patient may include a mammal, such as a mouse or a human.

[0018] In the present invention, the dosage of dapoxetine can be determined according to the patient's condition. Taking mice as an example, the dosage of dapoxetine can be 50-150 mg / kg body weight / day.

[0019] Dapanoxanil can improve tissue pathological damage, promote intestinal crypt cell regeneration, inhibit intestinal cell apoptosis, protect intestinal cell pyroptosis and reduce serum inflammatory factor levels. Therefore, Dapanoxanil can be used as a potential drug for the prevention and / or treatment of radiation-induced intestinal injury. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The figure shows the results of dapoxetine improving the pathological damage of intestinal tissue in radiation-induced intestinal injury;

[0021] Figure 2 The figure shows the results of dapoxetine promoting the regeneration of intestinal crypt cells in radiation-damaged intestine;

[0022] Figure 3 Shown is the result of dapoxetine inhibiting intestinal cell apoptosis in radiation-induced intestinal injury;

[0023] Figure 4 The figure shows the results of dapoxetine protecting intestinal cells from pyroptosis in radiation-induced intestinal injury;

[0024] Figure 5 The figure shows the results of dapoxetine reducing the secretion of serum inflammatory factors in radiation-induced intestinal injury. DETAILED DESCRIPTION

[0025] The present invention will be described in detail below by way of examples. In the following examples, the number of mice in each treatment group is 6.

[0026] Example 1

[0027] C57BL / 6J male mice aged about 8-10 weeks were placed in the abdomen 60 The radiation intestinal injury model was established by Co irradiation of 14Gy. Specifically: 2 hours before and 6 hours after Co source irradiation, the mice injected with dapamsulin (OLT1177) and the mice injected with dapamsulin after irradiation (IR+OLT1177) were intraperitoneally injected with 100mg / kg of dapamsulin solution, and the control group (NC) and the irradiation group (IR+NC) were injected with the same dose of solvent, followed by intraperitoneal injection once a day at the same time, and the modeling time was 4 days. Then the mice were treated as follows:

[0028] (1) Dapanoxanil improves pathological damage of intestinal tissue in radiation-induced intestinal injury

[0029] Intestinal villus length and crypt depth are the gold standards for reflecting intestinal absorption function. In order to evaluate the protective effect of dapoxetine on radiation-induced intestinal injury, intestinal tissues were collected 4 days after radiation, and HE staining (Solebol Hematoxylin and Eosin (HE) Staining Kit, G1120) was performed on them. They were observed and images were collected under an optical microscope. The results showed that the intestinal structure of mice in the NC group and the OLT1177 group was intact, the intestinal villi were neatly arranged and complete in morphology, and the crypt size outline was clearly discernible without obvious loss. 4 days after irradiation, the intestinal mucosal structure of mice in the IR group was obviously damaged, such as the intestinal villi shedding and blunting, and the number of intestinal crypts was significantly reduced. Compared with the IR group, the intestinal villus structure of mice in the IR+OLT1177 group was relatively complete, and the crypt depth was deepened. Subsequently, Image J software was used to measure and analyze the intestinal villus length. The villus length of mice in the IR+OLT1177 group almost returned to normal height ( Figure 1 The above results indicate that dapanoxic acid can improve the pathological damage of intestinal tissue in radiation-induced intestinal injury.

[0030] (2) Dapanoxanil promotes the regeneration of intestinal crypt cells in radiation-induced intestinal injury

[0031] After the intestine is damaged, intestinal stem cells located at the bottom of the intestinal crypts proliferate in large numbers, promoting the regeneration and repair of the intestinal mucosa. In order to evaluate the effect of dapamoate on the repair function of radiation-induced intestinal damage, BrdU staining was used to evaluate the regeneration capacity of crypt cells. BrdU is a thymidine nucleoside analog that can replace thymine (T) to infiltrate replicating DNA molecules during cell proliferation. The BrdU infiltration experiment can be used to observe the proliferation of intestinal crypt cells in mice after irradiation. After 4 days of radiation, BrdU 100 mg / kg was injected intraperitoneally, and intestinal tissue was collected 1 hour later for BrdU immunohistochemical staining (MedChemExpress, #HY-15910 ) , observed and collected images under an optical microscope. The results showed that there was no significant difference in the number of BrdU-positive crypt cells between the NC group and the OLT1177 group; on the 4th day after irradiation, the number of BrdU-positive crypt cells in the IR group decreased significantly, while the number of BrdU-positive crypt cells in the IR+OLT1177 group increased significantly ( Figure 2 The above results indicate that dapoxetine promotes the regeneration of intestinal crypt cells in radiation-induced intestinal injury.

[0032] (3) Dapanoxanil inhibits early intestinal cell apoptosis in radiation-induced intestinal injury

[0033] Radiation-induced apoptosis of intestinal cells is divided into two pathways: P53-PUMA-dependent and P53-independent, which mainly occur within 6-24 hours after irradiation. Terminal deoxynucleotidyl transferase dUTP nick end (TUNEL) marks apoptotic cell nuclei and has been widely used in the study of apoptosis. Caspase-3 is one of the key executors of apoptosis. Under normal circumstances, Caspase-3 in the cytoplasm exists as an inactive zymogen. The appearance of apoptotic signals can cause Caspase-3 to be cleaved and activated under the action of a variety of proteolytic enzymes. After 12 hours of irradiation, intestinal tissues were collected, and Western blotting was used to detect the expression of Caspase-3 protein in intestinal tissues. TUNEL immunohistochemistry staining (Boster Biological TUNEL Cell Apoptosis Two-Step Detection Kit (POD)) was observed and images were collected under an optical microscope. The results showed that after 12 hours of irradiation, the apoptosis rate of intestinal cells in the IR group increased significantly, and the expression of Caspase-3 was enhanced, while the apoptosis rate of IR+OLT1177 was significantly reduced, and the expression of Caspase-3 was weak ( Figure 3 , "****" indicates p<0.0001). The above results indicate that dapanoxacin inhibits the apoptosis of intestinal cells in the early stage of radiation-induced intestinal injury.

[0034] (4) Dapanoxanil protects intestinal cells from pyroptosis in radiation-induced intestinal injury

[0035] Irradiation activates inflammasomes in multiple ways and pathways, which in turn cleaves GSDMD under the action of the effector protein Caspase-1, ultimately leading to cell pyroptosis. After 4 days of irradiation, intestinal tissues were collected, and Western blotting was used to detect NLRP3 inflammasome activation and cell pyroptosis in the intestinal tissues of mice. The results showed that compared with the NC group, the spliceosome levels of NLRP3, Caspase-1, and GSDMD in the intestinal tissues of mice in the IR group were significantly upregulated, while the activation of Caspase-1 and GSDMD proteins in the intestinal tissues of the IR+OLT1177 group was significantly inhibited compared with the IR group ( Figure 4 ). The above results indicate that dapanoxacil can protect intestinal cells from pyroptosis in radiation-induced intestinal injury.

[0036] (5) Dapanoxanil reduces the secretion of serum inflammatory factors in radiation-induced intestinal injury

[0037] After radiation-induced intestinal damage, the body produces a large amount of PAMPs and DAMPs, which activate the immune system and cause a significant increase in the level of inflammatory factors in the serum. Before collecting the mouse intestinal tissue in step (1), the mouse serum samples were collected, and the micro-sample multi-index flow protein quantitative technology (Cytometric Bead Array, CBA) was used to detect the effect of dapoxetine on the secretion of inflammatory factors in the mouse systemic serum. The results showed that the levels of TNF-α, IL-6, MCP-1, IL-10, and IFN-γ in the serum of the IR group were significantly increased compared with those of the NC group (P<0.0001), and the levels of TNF-α, IL-6, MCP-1, IL-10, and IFN-γ in the serum of the IR+OLT1177 group were significantly decreased compared with those of the IR group ( Figure 5 "**" indicates P<0.01, "****" indicates P<0.0001, "*" indicates P<0.05, and "***" indicates P<0.001.) The above results indicate that dapanoxic acid reduces the secretion of serum inflammatory factors in radiation-induced intestinal injury.

[0038] The preferred embodiments of the present invention are described in detail above, but the present invention is not limited thereto. Within the technical concept of the present invention, the technical solution of the present invention can be subjected to a variety of simple modifications, including the combination of various technical features in any other suitable manner, and these simple modifications and combinations should also be regarded as the contents disclosed by the present invention and belong to the protection scope of the present invention.

Claims

1. Use of dapanoxacil in the preparation of a drug for preventing and / or treating radiation-induced intestinal injury induced by a local irradiation model of ionizing radiation.

2. The use according to claim 1, wherein: The radiation intestinal injury is intestinal injury caused by local irradiation of ionizing radiation.

3. The use according to claim 1, wherein: The radiation-induced intestinal injury is a complication caused by radiotherapy for malignant tumors in the abdomen and / or pelvis.

4. The use according to claim 3, wherein: The malignant tumor is at least one of ovarian malignant tumor, endometrial cancer, cervical cancer and pelvic malignant lymphoma.

5. The use according to claim 3 or 4, wherein: The ionizing radiation energy absorption dose of the radiotherapy is 12-16Gy / time.

6. Use of dapanoxacin in the preparation of drugs for improving pathological damage of intestinal tissue.

7. Use of dapanoxacin in the preparation of a drug for promoting the regeneration of intestinal crypt cells.

8. Use of dapanoxacin in the preparation of drugs for inhibiting intestinal cell apoptosis or pyroptosis.

9. Use of dapanoxacin in the preparation of a drug for reducing the secretion of serum inflammatory factors.

10. The use according to claim 9, wherein: The inflammatory factor is selected from at least one of TNF-α, IL-6, MCP-1, IL-10 and IFN-γ.

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