Application of diacetamide trinitrogen amidine in preparation of medicine for preventing, relieving or treating intestinal injury diseases caused by ionizing radiation

Abstract

The invention relates to the technical field of medicines, in particular to application of diacetamide trinitrogen amidine in preparation of medicines for preventing, relieving or treating intestinal injury diseases caused by ionizing radiation. It is found for the first time that diacetamide triazamidine can effectively relieve intestinal injury caused by ionizing radiation and does not reduce the radiosensitivity of tumor cells, the compound can protect normal intestinal tissue, and meanwhile the limitation that a radiation protection medicine can possibly protect tumors is overcome; the invention provides a new treatment scheme for radioactive intestinal injury of organisms, and can be used for preventing, relieving or treating radioactive intestinal injury caused by radiotherapy. The invention develops new application of the diacetamide trinitrogen amidine and has a wide application prospect.

Description

Technical Field

[0001] The invention relates to the field of pharmaceutical technology, and in particular to application of diethylamide triazoline in the preparation of medicines for preventing, alleviating or treating intestinal damage diseases caused by ionizing radiation. Background Art

[0002] Radiation intestinal injury, also known as radiation enteropathy, is the most common and most serious complication after radiotherapy for abdominal and pelvic tumors. Statistics show that almost all patients undergoing radiotherapy for abdominal and pelvic tumors will experience obvious gastrointestinal symptoms, of which 15%-20% of patients have severe symptoms and need to change their treatment plans. Radiation intestinal injury is still the most important limiting factor for current radiotherapy doses. In addition, the widespread application of nuclear technology in the fields of energy and medicine has provided important guarantees for national energy security and national health. At the same time, however, nuclear power plant accidents have occurred many times internationally, and the widespread popularization of medical radiation devices may also lead to medical accidents, all of which may cause radiation intestinal injury.

[0003] Radiation intestinal injury can be divided into two stages: acute radiation enteritis occurs within a few days after radiotherapy, mainly due to the mutual promotion of mucosal barrier destruction caused by insufficient intestinal epithelial renewal and cascade-amplified inflammation, clinically manifested as nausea, abdominal pain, diarrhea and infection, and in severe cases can lead to septic shock; chronic (delayed) enteritis occurs several months to years after radiation exposure, and its pathological characteristics are irreversible intestinal villous atrophy, intestinal wall fibrosis and microvascular sclerosis, clinically manifested as malnutrition, chronic diarrhea, and may progress to intestinal obstruction and intestinal perforation. At present, the treatment of radiation intestinal injury is mainly symptomatic, including the use of hemostatic drugs, antibiotics, steroid drugs, intestinal mucosal protective agents, probiotics, parenteral nutrition support, etc. to control intestinal symptoms, etc. Complications caused by delayed enteritis such as obstruction, bleeding, perforation, stenosis, ulcers, etc. are mainly solved through surgery. Amifostine (WR2721) is currently the only specific drug approved for reducing radiation damage. However, due to its serious adverse reactions (nausea, vomiting, hypotension, etc.) and narrow therapeutic window, it is only approved for use in patients with head and neck cancer undergoing radiotherapy to reduce the incidence of moderate to severe dry mouth symptoms, and has not been approved for radiation-induced intestinal injury. In short, there are currently no specific preventive and therapeutic drugs for radiation-induced intestinal injury in clinical practice, and some treatment methods lack evidence-based evidence, and lack effective and reliable standardized treatment plans. The intestinal radiation damage protective agents under development focus on promoting DNA damage repair or reducing oxidative stress levels, which are not very specific and may protect tumor cells. In addition, there are problems such as large side effects and high prices. The prospects for new drug development are still unclear. With the improvement of cancer diagnosis and treatment, the survival time of patients has been prolonged, and the number of cancer survivors has increased significantly. The importance of reducing the side effects of radiotherapy and improving the quality of life of patients has become increasingly prominent. Therefore, it is of great significance to find drugs to prevent and treat radiation enteritis.

[0004] Diminazene Aceturate, also known as Benil and Xuechongjing, is a well-tolerated antiparasitic drug mainly used to treat trypanosomiasis in livestock. The compound was launched in 1955. It is currently believed that its mechanism of action is to selectively block the DNA synthesis or replication of the trypanosome kinetoplast and to irreversibly bind to the cell nucleus, thereby causing the kinetoplast of the trypanosome to disappear and prevent it from dividing and reproducing. Studies in recent years have shown that diminazene Aceturate is a specific agonist of angiotensin-converting enzyme 2 (ACE2), thereby activating the renin-angiotensin system and inducing the cleavage of angiotensin II (Ang II) into angiotensin 1-7 (Ang 1-7), thereby counteracting the physiological effects of Ang II such as vasoconstriction, pro-inflammation, pro-coagulation, and fibrosis. Summary of the invention

[0005] The present invention has found for the first time through experimental verification that diethylamide triazoline can effectively reduce intestinal damage caused by ionizing radiation without reducing the radiation sensitivity of tumor cells. Based on this, the following technical solution is proposed.

[0006] First, the present invention provides the use of diethylamide triazoline in the preparation of medicines; the medicines are used in at least one of the following aspects: (1) Prevent, alleviate or treat intestinal damage caused by ionizing radiation, intestinal villus damage, intestinal barrier destruction or inflammatory cell infiltration; (2) Reduce the death of intestinal organoids caused by ionizing radiation; (3) Radioprotection of intestinal organoids.

[0007] Preferably, the intestinal damage disease caused by ionizing radiation is radiation enteritis.

[0008] In some embodiments, the radiation enteritis is caused by radiation therapy for pelvic malignancies and / or abdominal malignancies and / or retroperitoneal malignancies.

[0009] In some embodiments, the radiation enteritis results from a medical accident.

[0010] In some embodiments, the drug comprises diethylamide triazoline and pharmaceutically acceptable excipients.

[0011] In some embodiments, the pharmaceutically acceptable excipients include fillers, excipients, lubricants, wetting agents, diluents and the like.

[0012] In some embodiments, the dosage form of the drug is a solid preparation or a liquid preparation.

[0013] Preferably, the solid preparation is a powder, granule, capsule or tablet, and the liquid preparation is an oral solution or injection.

[0014] In the above application, the drug does not reduce the radiation sensitivity of intestinal tumor cells.

[0015] In the above application, the drug achieves the application purpose by increasing the survival rate after lethal dose irradiation.

[0016] In the above application, the drug achieves the application purpose through at least one of the following ways: (1) Inhibit irradiation-induced apoptosis of small intestinal crypt cells; (2) Promote the proliferation of irradiated small intestinal crypt cells; (3) Promote the recovery of intestinal villi; (4) Inhibit irradiation-induced inflammatory cell infiltration and pro-inflammatory cytokine levels in small intestinal tissue.

[0017] In the above application, the drug achieves the application purpose by inhibiting the degradation of intestinal tight junction proteins caused by radiation and maintaining the integrity of the intestinal epithelial barrier.

[0018] Compared with the prior art, the present invention has the following beneficial effects: The present invention discovers for the first time that diethylamide triazoline can effectively reduce intestinal damage caused by ionizing radiation without reducing the radiation sensitivity of tumor cells. The compound can protect normal intestinal tissue while overcoming the limitation that radiation protection drugs may protect tumors, providing a new treatment plan for radiation intestinal damage in organisms, and can be used to prevent, alleviate or treat radiation intestinal damage caused by radiotherapy. The present invention has opened up new uses for diethylamide triazoline and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The effect of diethylamide triazine on the survival of mice irradiated with 14 Gy whole body irradiation combined with bone marrow transplantation.

[0020] Figure 2 The effect of diethylamide triazine on apoptosis of small intestinal crypt cells after 14Gy irradiation.

[0021] Figure 3 The effect of diethylamide triazine on the length of small intestinal villi after 14Gy irradiation.

[0022] Figure 4 The effect of diethylamide triazine on the proliferation of small intestinal crypt cells after 14Gy irradiation.

[0023] Figure 5 The effect of diethylamide triazine on neutrophil infiltration in the small intestine after 14Gy irradiation.

[0024] Figure 6The effect of diethylamide triazine on the expression of IL-1β, TNF-α, and IL-6 proteins in small intestinal tissue after 14Gy irradiation.

[0025] Figure 7 The effect of diacetylamide triazine on the expression of tight junction proteins Occludin and ZO-1 in small intestinal epithelial cells after 14Gy irradiation.

[0026] Figure 8 The effect of diethylamide triazine on the growth of small intestinal organoids after 12 Gy irradiation.

[0027] Fig. 9 The effect of diethylamide triazine on the growth of colon cancer cell lines after 12Gy irradiation.

[0028] Fig.10 This is the effect of diethylamide triazine on the growth of colorectal tumors after radiotherapy; the left picture is the endoscopic image of tumor growth after different treatments in the in situ tumor model, and the right picture is the quantitative results of tumors of different diameters. DETAILED DESCRIPTION

[0029] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0030] In the examples provided in this specification, if no specific techniques or conditions are specified, the techniques or conditions described in the literature in this field or the product instructions are used. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased through regular channels.

[0031] The CAS number of diethylamide triazoline in this specification is 908-54-3.

[0032] Example 1: Diethylamide triazoline improves survival rate after lethal dose irradiation 1.1 Experimental animals: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g.

[0033] 1.2 Experimental groups: The mice were divided into irradiation control group (IR) and diethylamide triazepam group (IR+DIZE), with 10 mice in each group.

[0034] 1.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours and 2 hours before irradiation, at a dose of 50 mg / kg, by intraperitoneal injection.

[0035] 1.4 Experimental steps: (1) The drug-treated group was given drugs before irradiation, and the control group was given normal saline; (2) The recipient mice received whole-body X-ray irradiation with a dose of 14 Gy and a dose rate of 0.72 Gy / min; (3) 6 hours after irradiation, unirradiated donor mice were obtained and bone marrow single cell suspension was prepared under sterile conditions; (4) Bone marrow transplantation: 5×10 single cell suspension of the donor mouse bone marrow was used. 6 / mice were injected into the tail vein of recipient mice; (5) After bone marrow transplantation, the recipient mice were returned to their cages, allowed to eat freely, and their survival was observed within 30 days.

[0036] 1.5 Experimental results: like Figure 1 As shown, all mice in the control group died within 17 days, with an average survival time of 8.6 days. After administration of diethylamide triazoline, the survival rate increased to 70% within 30 days, and the difference was statistically significant ( p =0.0038).

[0037] Example 2: Diethylamide triazoline inhibits irradiation-induced apoptosis of small intestinal crypt cells 2.1 Experimental animals: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g.

[0038] 2.2 Experimental groups: The mice were divided into irradiation control group (IR) and diethylamide triazene group (IR+DIZE).

[0039] 2.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours and 2 hours before irradiation, with a dose of 50 mg / kg, by intraperitoneal injection.

[0040] 2.4 Experimental steps: (1) The drug-treated group was given drugs before irradiation, and the control group was given normal saline; (2) The recipient mice received whole-body X-ray irradiation with a dose of 14 Gy and a dose rate of 0.72 Gy / min; (3) 4 hours and 24 hours after irradiation, small intestine jejunum segments were obtained, fixed with formaldehyde, dehydrated, and embedded in paraffin; (4) Prepare paraffin sections of intestinal tissue with a thickness of 4 μm; (5) TUNEL kit was used to detect cell apoptosis in paraffin sections of intestinal tissue. Five intestinal loops were counted under a microscope for each mouse, and the number of apoptotic cells in five complete crypts in each intestinal loop was counted.

[0041] 2.5 Experimental results: like Figure 2 As shown, a large number of mouse intestinal crypt cells underwent apoptosis 4 hours and 24 hours after irradiation, mainly distributed in the rapid expansion area and the bottom of the crypt, with (6.7±2.3) cells / crypt and (5.1±1.3) cells / crypt, respectively. The number of apoptotic cells in the diethylamide triazine group decreased by 60% and 30% at the above two time points, respectively, and the difference was statistically significant.

[0042] Example 3 Diethylamide triazoline promotes the proliferation of small intestinal crypt cells and the recovery of intestinal villi after irradiation 3.1 Experimental animals: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g.

[0043] 3.2 Experimental groups: The mice were divided into irradiation control group (IR) and diethylamide triazene group (IR+DIZE).

[0044] 3.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours and 2 hours before irradiation, with a dose of 50 mg / kg, by intraperitoneal injection.

[0045] 3.4 Experimental steps: (1) The drug-treated group was given drugs before irradiation, and the control group was given normal saline; (2) The recipient mice received whole-body X-ray irradiation with a dose of 14 Gy and a dose rate of 0.72 Gy / min; (3) 5-Fluorouracil (BrdU) was intraperitoneally administered at 120 mg / kg 2 hours before sampling. The sampling time points were before irradiation and 24, 48, 84, and 120 hours after irradiation. The small intestine jejunum segments were obtained and fixed with formaldehyde; (4) Prepare paraffin sections of intestinal tissue with a thickness of 4 μm; (5) Measurement of intestinal villus length: paraffin sections of intestinal tissue were taken and dewaxed and stained with HE. The length of complete intestinal villi in the intestinal cavity was measured under a microscope. Five intestinal loops were counted for each mouse, and the length of five complete villi was counted for each intestinal loop. (6) Mouse crypt cell proliferation detection: paraffin sections of intestinal tissue were dewaxed and stained with BrdU immunohistochemistry. The number of positive cells in the crypts was counted under a microscope. Five intestinal loops were counted for each mouse, and the number of positive cells in five well-shaped crypts in each intestinal loop was counted.

[0046] 3.5 Experimental results: like Figure 3As shown in the figure, the length of intestinal villi of mice in the IR group gradually shortened after irradiation, and the density of intestinal epithelial cells on the surface of the villi changed from dense to sparse. The quantitative results showed that the average length of the villi was (577.98±51.4) μm when not irradiated, and the average length of the villi was shortened to (551.28±78.15) μm, (529.23±101.52) μm, (468.31±81.32) μm, and (475.51±81.69) μm at 24, 48, 84, and 120 hours after irradiation, respectively. There was no significant difference between the villi length of the drug-treated group and the control group before irradiation, and the degree of shortening of the villi length was significantly reduced after irradiation, which was 119%, 127%, 141%, and 158% of the corresponding time points after irradiation, respectively.

[0047] like Figure 4 As shown in the figure, the number of proliferating cells in the intestinal crypts of the IR group decreased rapidly. At 24, 48, and 84 hours after irradiation, the average number of proliferating cells in each crypt decreased from (14.73±2.69) before irradiation to (3.39±2.69), (2.14±1.14), and (13.5±6.44), respectively. There was no significant difference in the proliferation of cells in the crypts of the drug-treated group before irradiation compared with the control group. Correspondingly, the number of proliferating cells at each time point after irradiation was (5.36±1.38) / crypt, (2.69±1.15) / crypt, and (17.69±6.54) / crypt, with statistically significant differences.

[0048] Example 4: Diethylamide triazoline inhibits irradiation-induced inflammatory cell infiltration in small intestinal tissue and elevated levels of pro-inflammatory cytokines 4.1 Experimental animals: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g.

[0049] 4.2 Experimental groups: The mice were divided into normal control group (Vehicle), simple drug administration group (DIZE), irradiation control group (IR) and diethylamide triazine group (IR+DIZE).

[0050] 4.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours and 2 hours before irradiation, with a dose of 50 mg / kg, by intraperitoneal injection.

[0051] 4.4 Experimental steps: (1) The drug-treated group was given drugs before irradiation, and the control group was given normal saline; (2) The recipient mice received whole-body X-ray irradiation with a dose of 14 Gy and a dose rate of 0.72 Gy / min; (3) Ly6G immunohistochemistry to detect neutrophil infiltration in small intestinal tissue: Small intestinal tissue was obtained 48 hours after irradiation, fixed with formaldehyde and then made into paraffin sections with a thickness of 4 μm. After dewaxing, the sections were subjected to Ly6G immunohistochemistry to label neutrophils. The number of positive cells in the field of view was counted under a 20x objective lens. 25 fields of view were counted for each mouse. (4) Western Blot detection of the expression of inflammatory factors (IL-1β, IL-6, TNF-α) in small intestinal tissue: 48 hours after irradiation, small intestinal tissue was obtained, the tissue was homogenized with lysis buffer, and the protein was extracted. Then, the protein concentration was determined by the BCA method to adjust the sample and perform SDS-PAGE electrophoresis. After electrophoresis, the protein was transferred from the gel to a PVDF membrane and blocked at room temperature. Then, it was incubated with specific IL-1β, IL-6, and TNF-α primary antibodies at 4°C overnight, and then incubated with secondary antibodies at room temperature for 1 hour. The signal was detected by fluorescence method, and the protein expression was calculated by gray value.

[0052] 4.5 Experimental results: like Figure 5 As shown in the data, under non-irradiation conditions, there was little neutrophil infiltration in the intestinal tissue propria in the control and simple drug administration groups. After irradiation, the number of Ly6G-positive cells increased by 60 times, with the number being approximately (20.66±4.53) / field of view. The number in the IR+DIZE group decreased to (7.72±2.20) / field of view, and the difference was statistically significant.

[0053] like Figure 6 As shown, IL-1β, IL-6, and TNF-α in tissues increased significantly after irradiation, while the administration of diethylamide triazoline significantly inhibited the expression of various inflammatory factors.

[0054] Example 5: Diethylamide triazoline inhibits the degradation of intestinal tight junction proteins caused by irradiation and maintains the integrity of the intestinal epithelial barrier 5.1 Experimental animals: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g.

[0055] 5.2 Experimental groups: The mice were divided into normal control group (Vehicle), simple drug administration group (DIZE), irradiation control group (IR) and diethylamide triazine group (IR+DIZE).

[0056] 5.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours and 2 hours before irradiation, with a dose of 50 mg / kg, by intraperitoneal injection.

[0057] 5.4 Experimental steps: (1) The drug-treated group was given drugs before irradiation, and the control group was given normal saline; (2) The recipient mice received whole-body X-ray irradiation with a dose of 14 Gy and a dose rate of 0.72 Gy / min; (3) Occludin and ZO-1 immunofluorescence detection of tight junction proteins in small intestinal epithelial cells: The small intestinal tissue was obtained 48 hours after irradiation, fixed with formaldehyde and then paraffin sections were made. The section thickness was 4 μm. After dewaxing, the sections were immunofluorescently labeled with occludin and ZO-1 for tight junction proteins. The fluorescence microscope was used to take pictures and the fluorescence quantification was performed using Image J software.

[0058] 5.5 Experimental results: like Figure 7 As shown in the data, tight junction protein Occludin is mainly distributed in the brush border of small intestinal epithelial cells, and tight junction protein ZO-1 is mainly distributed in the junctions between small intestinal epithelial cells. Simple administration of diethylamide triazine does not affect the expression of the two. Compared with the normal control, the expression of Occludin and ZO-1 in the IR group decreased by 21% and 33%, respectively, while the corresponding (IR+DIZE) group only decreased by 17% and 19%. There was a statistically significant difference between the IR+DIZE group and the IR group.

[0059] Example 6: Diethylamide triazoline has a radioprotective effect on in vitro irradiated intestinal organoids 6.1 Experimental animals and main reagents: C57BL / 6J mice, male, 6-8 weeks old, weighing 20-22 g, intestinal organoid culture medium (MethoCult GF M3434, Stemcell).

[0060] 6.2 Experimental groups: The mice were divided into normal control group (Vehicle), simple drug administration group (DIZE), irradiation control group (IR) and diethylamide triazine group (IR+DIZE).

[0061] 6.3 Experimental steps: (1) Obtaining small intestinal crypts: After killing the mice, 8-10 cm of jejunum were obtained under sterile conditions, and the extraintestinal mesentery, blood vessels, and fat were removed. After longitudinally cutting the intestine, the intestinal villi were gently scraped off, and the intestine was cut into approximately 1 cm fragments with a blade. The intestinal fragments were washed with ice-cold DPBS for 6-8 times, and ice-cold 2mM EDTA / DPBS solution was added. Incubate on ice for 20 minutes, shake vigorously for 30 seconds, collect the liquid, filter with a 70μm filter, and observe the purity of the complete crypts under a microscope; (2) Small intestinal organoid culture: Centrifuge the prepared crypt solution at 150g for 3 min at 4°C, discard the supernatant, resuspend the precipitate in pre-cooled DMEM / F-12, count, dilute to an appropriate concentration for inoculation, mix with the prepared matrix gel at a ratio of 1:1, and then inoculate 50 μL / well into a 24-well plate. Add 100 μL of organoid culture medium and culture at 37°C with 5% CO2. Observe the growth of the organoids daily. (3) After the organoids have grown stably, they are passaged, randomly divided into groups, and given 2 μM diethylamide triazoline; (4) The grouped organoids were irradiated with 12 Gy of X-rays at a dose rate of approximately 72 cGy / min; (5) Eight days after irradiation, record the organoid survival rate, number of buds, and average surface area.

[0062] 6.4 Experimental Results like Figure 8 As shown, the survival rate, number of buds and surface area of ​​organoids decreased significantly after irradiation. After the administration of diethylamide triazoline, the survival rate, number of buds and surface area increased by 14%, 20% and 50%, respectively, and the difference was statistically significant.

[0063] Example 7 Diethylamide triazoline does not reduce the radiation sensitivity of colon tumor cells 7.1 Experimental cells: mouse colon cancer cell line (CT-26), human colon adenocarcinoma cell line (RKO), human colon adenocarcinoma cell line (HT-29).

[0064] 7.2 Experimental groups: divided into control group (CTR), simple drug administration group (DIZE), irradiation control group (IR) and irradiation drug administration group (IR+DIZE).

[0065] 7.3 Experimental steps: (1) Plate tumor cells in a 96-well plate, 1000 cells / well; (2) After the cells adhered to the wall, diethylamide triazoline was added to the drug group at a final concentration of 2 μM, and physiological saline was added to the CTR and IR groups; (3) 24 h after administration, cells were irradiated with 12 Gy of X-rays at a dose rate of approximately 72 cGy / min; (4) Use the CCK8 method to detect cell viability daily, calculate the cell number, and draw a growth curve.

[0066] 7.4 Experimental Results like Fig. 9As shown, under non-irradiation conditions, no significant promoting effect on the growth of tumor cells was observed after treatment with diethylamide triazoline, while cell growth was inhibited after irradiation. Pretreatment with diethylamide triazoline did not significantly promote or inhibit cell growth.

[0067] Example 8 Diethylamide triazoline does not reduce the sensitivity of colorectal tumors to radiotherapy 8.1 Experimental animals: C57BL / 6J mice, male, 4-6 weeks old, weighing 18-20 g.

[0068] 8.2 Experimental groups: The mice were divided into normal control group (Vehicle), simple drug administration group (DIZE), irradiation control group (IR) and diethylamide triazine group (IR+DIZE).

[0069] 8.3 Administration method: Diethylamide triazoline was dissolved in physiological saline and used immediately after preparation. The recipient mice were given the drug twice, 24 hours before each irradiation, at a dose of 15 mg / kg, by intraperitoneal injection.

[0070] 8.4 Experimental steps: (1) Dextran sulfate sodium (DSS) and azoxymethane (AOM)-induced colon cancer model: After intraperitoneal injection of 10 mg / kg AOM, mice were fed with drinking water containing 2% DSS for 5 consecutive days, followed by a 2-day rest period in which they were fed with drinking water without DSS. This cycle was repeated four times in total. After the last cycle, the model was considered successful when tumor formation was confirmed by small animal endoscopy. (2) The mice with successful modeling were randomly divided into groups, with 6 mice in each group; (3) Mice were treated with medication and / or irradiation according to the group. The irradiation conditions were as follows: local X-ray irradiation of the lower abdomen of mice, 2 Gy each time, for 5 consecutive days. The medication conditions were intraperitoneal injection of diethylamide triazoline 15 mg / kg, 2 hours before each irradiation. (4) Endoscopic observation of the tumor after radiotherapy; (5) The mice were killed and the number and diameter of tumors were counted under a stereomicroscope.

[0071] 8.5 Experimental results: like Fig.10As shown in the figure, scattered multiple intestinal tumors can be seen in the colon of mice that did not receive radiotherapy. The tumor volume was large, accounting for about 1 / 3-1 / 2 of the intestinal wall. After simple administration of diethylamide triazoline, there was no significant change in tumor growth. The tumor volume was significantly reduced after radiotherapy, and no significant effect on the radiotherapy effect was observed after administration. Mice were killed after radiotherapy, and the number and size of tumors were counted. The results were consistent with those of endoscopy. The number of tumors in the CTR group was about (7.40±2.5), of which the proportions of tumors with diameters less than 2mm, between 2mm-5mm, and greater than 5mm were 32.20%, 61.01%, and 6.78%, respectively. There was no significant difference in the number and size of tumors in the DIZE group compared with the IR group. After radiotherapy, the number of colorectal tumors decreased to (4.1±2.4), and all tumors with a diameter of more than 5mm disappeared, indicating that radiotherapy had a killing effect on tumors. There was no significant difference in the number and size of tumors in the (IR+DIZE) group compared with the IR group.

[0072] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of diethylamide triazoline in the preparation of medicines; the medicines are used in at least one of the following aspects: (1) Prevent, alleviate or treat intestinal damage caused by ionizing radiation, intestinal villus damage, intestinal barrier destruction or inflammatory cell infiltration; (2) Reduce the death of intestinal organoids caused by ionizing radiation; (3) Radioprotection of intestinal organoids.

2. The use according to claim 1, characterized in that: The intestinal damage disease caused by ionizing radiation is radiation enteritis.

3. The use according to claim 2, characterized in that: The radiation enteritis is caused by radiotherapy of pelvic malignant tumors and / or abdominal malignant tumors and / or retroperitoneal malignant tumors.

4. The use according to claim 1, characterized in that: The medicine comprises diethylamide triazoline and pharmaceutically acceptable auxiliary materials.

5. The use according to claim 1, characterized in that: The dosage form of the drug is a solid preparation or a liquid preparation.

6. The use according to claim 5, characterized in that: The solid preparation is powder, granule, capsule or tablet, and the liquid preparation is oral solution or injection.

7. The use according to any one of claims 1 to 6, characterized in that: The drug does not reduce the radiosensitivity of intestinal tumor cells.

8. The use according to claim 1, characterized in that: The medicine achieves its application purpose by improving the survival rate after irradiation with a lethal dose.

9. The use according to claim 1, characterized in that: The drug achieves its application purpose through at least one of the following approaches: (1) Inhibit irradiation-induced apoptosis of small intestinal crypt cells; (2) Promote the proliferation of irradiated small intestinal crypt cells; (3) Promote the recovery of intestinal villi; (4) Inhibit irradiation-induced inflammatory cell infiltration and pro-inflammatory cytokine levels in small intestinal tissue.

10. The use according to claim 1, characterized in that: The drug achieves its application purpose by inhibiting the degradation of intestinal tight junction proteins caused by radiation and maintaining the integrity of the intestinal epithelial barrier.

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