Application of dapanoxacin in the prevention and / or treatment of radiation-induced intestinal injury

Abstract

The present invention relates to the fields of biology and medicine and discloses the use of dapamuxil in preventing and / or treating radiation-induced intestinal injury. The inventors of the present invention have discovered that dapamuxil protects against radiation-induced intestinal injury induced by localized ionizing radiation exposure by ameliorating histopathological damage, promoting intestinal crypt cell regeneration, inhibiting intestinal cell apoptosis, ameliorating intestinal cell pyroptosis, and reducing serum inflammatory factor levels. Therefore, it is a potential drug for preventing and / or treating radiation-induced intestinal injury.

Description

Technical Field

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

[0002] Radiation-induced bowel injury (RABI) is an intestinal complication caused by radiotherapy for malignant tumors in the pelvis, abdomen, and other areas. Lesions can affect the small intestine, rectum, and colon. Symptoms include abdominal pain, diarrhea, and bloody stools. Severe cases may develop complications such as intestinal ulcers and even perforation. Acute RBI is a key factor in mortality and treatment failure associated with systemic radiation injury, and represents a bottleneck in the successful treatment of extremely severe radiation sickness. Currently, approximately 50% of cancer survivors receive radiotherapy for intra-abdominal and pelvic cancers, and the global burden of RBI is expected to increase significantly in the future. Although extensive research has been conducted on the prevention and treatment of ARBBI, many drug, biological, and cell-based therapies remain in the preclinical stage, and currently, no specific therapeutic agents exist to treat or prevent ARBBI. Therefore, developing new strategies and drugs for the prevention and treatment of ARBBI is of great importance.

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

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

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

[0006] Dapanoxonitrile is 3-methanesulfonylpropionitrile, 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, which are intestinal damage caused by local irradiation of ionizing radiation. The radiation intestinal injury is preferably intestinal damage 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 patients with malignant tumors. 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-induced 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 dapanoxacin in the preparation of a medicament for improving pathological damage of intestinal tissue caused by radiation (radiation-induced intestinal injury).

[0010] The third aspect of the present invention provides the use of dapamoic acid in the preparation of a medicament for promoting the regeneration of intestinal crypt cells (in radiation-induced intestinal damage).

[0011] A fourth aspect of the present invention provides the use of dapamoic acid in the preparation of a medicament for inhibiting (radiation-induced) intestinal cell apoptosis.

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

[0013] A sixth aspect of the present invention provides the use of dapamoic acid in the preparation of a medicament for reducing the secretion of serum inflammatory factors (particularly 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 of dapoxetine, but it is preferably at least one of intravenous injection, intramuscular injection, and subcutaneous injection. Those skilled in the art can formulate dapoxetine into a specific dosage form, such as an injection, according to the administration method.

[0017] In the present invention, the patient may include mammals, such as mice or humans.

[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 dapamoic acid improving the pathological damage of intestinal tissue in radiation-induced intestinal injury;

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

[0022] Figure 3 Shown are the results of dapamoic acid inhibiting intestinal cell apoptosis in radiation-induced intestinal injury;

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

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

[0025] The present invention will be described in detail below through 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 A radiation-induced intestinal injury model was established by irradiating 14 Gy of Co. Specifically, 2 hours before and 6 hours after irradiation with a cobalt source, mice injected with 100 mg / kg of dapamoxil solution were intraperitoneally injected into the dapamoxil group (OLT1177) and the irradiated dapamoxil group (IR+OLT1177). The same dose of solvent was injected into the control group (NC) and the irradiated group (IR+NC). Subsequently, the mice were intraperitoneally injected once a day at the same time for 4 days. The mice were then 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 dapamoate on radiation-induced intestinal injury, intestinal tissues were collected 4 days after irradiation, and HE staining (Solebol Hematoxylin and Eosin (HE) staining kit, G1120) was performed on them. They were observed under an optical microscope and images were collected. 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 the morphology was intact, and the crypt size outlines were clearly discernible and there was no obvious loss. 4 days after irradiation, the intestinal mucosal structure of mice in the IR group was obviously damaged, such as the shedding and blunting of intestinal villi and a significant decrease in the number of intestinal crypts. 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 dapanoxacin 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 massively, 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 regenerative 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, 100 mg / kg of BrdU was injected intraperitoneally, and intestinal tissue was collected 1 hour later for BrdU immunohistochemical staining (MedChemExpress, #HY-15910 ) Observe and collect 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 "**" indicates p < 0.01). The above results indicate that dapamoic acid 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 cell apoptosis. Under normal circumstances, Caspase-3 in the cytoplasm exists as an inactive zymogen. The appearance of apoptosis signals can lead to the cleavage and activation of Caspase-3 under the action of a variety of proteolytic enzymes. 12 hours after 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 used to observe and collect images under an optical microscope. The results showed that 12 hours after irradiation, the apoptosis rate of intestinal cells in the IR group was significantly increased, 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 The above results indicate that dapanoxacin inhibits the early apoptosis of intestinal cells in 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. Four days after 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 levels of NLRP3, Caspase-1, and GSDMD splicing 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 dapanoxacin 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 effect of dapoxetine on the secretion of inflammatory factors in the mouse systemic serum was detected using a micro-sample multi-index flow protein quantitative technology (Cytometric Bead Array, CBA). 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 dapanoxanil 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, various simple variations of the technical solution of the present invention may be made, including combining the various technical features in any other appropriate manner. These simple variations and combinations should also be regarded as the contents disclosed by the present invention and fall within the scope of protection of the present invention.

Claims

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

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