Compound for relieving cell injury caused by heavy ion radiation, medicine and application

Abstract

The invention relates to the technical field of biological medicine, and particularly provides a medicine for relieving cell damage caused by heavy ion radiation and application of the medicine. The medicine can obviously improve the cell cloning survival rate under the condition of carbon ion radiation, inhibit necroptosis and weaken inflammation-related transcription reaction caused by radiation. According to the present invention, the multi-level experiments such as clone formation experiment, protein level detection, transmission electron microscope observation, transcriptomics analysis and the like prove that the pharmaceutical composition can specifically improve the cell survival state after carbon ion irradiation, and has no significant protection effect on X-ray irradiation; the invention provides a new pharmacological scheme for heavy ion radiotherapy related protection and complication intervention, and has important application prospects.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical biotechnology, and in particular to a compound, drug, and application for alleviating cell damage caused by heavy ion radiation. Technical Background

[0002] Radiotherapy is an important treatment for malignant tumors, and heavy ion radiotherapy (such as carbon ion radiotherapy) is gradually being promoted globally due to its physical and biological advantages. Compared with traditional X-ray or proton radiotherapy, carbon ion radiotherapy has the following characteristics: ① Bragg peak effect: The way carbon ions deposit energy in tissues allows the dose to be precisely concentrated in the tumor target area, thereby reducing damage to surrounding normal tissues. ② High linear energy transfer (LET): Carbon ions have a high LET value, which can produce dense double-strand breaks on DNA molecules, thereby causing a stronger killing effect on tumor cells. ③ Low hypoxia enhancement ratio: Carbon ions are more effective at killing hypoxic cells than photon radiation, making them more suitable for treating hypoxic and radiation-resistant tumors.

[0003] However, while carbon ion radiation kills tumor cells, it also causes unique and severe damage to normal cells and tissues, mainly manifested in: 1. Induction of necroptosis, a cell death mechanism different from traditional apoptosis, which involves cell membrane rupture and release of cell contents, often triggering a severe inflammatory response; 2. Activation of inflammation-related signaling pathways, such as the NF-κB pathway, leading to the upregulation of pro-inflammatory factors (IL1A, IL1B, IL6, CXCL1 / 2 / 3, etc.), which may exacerbate tissue inflammation and adverse reactions. Currently, several radioprotective drugs or radiosensitizers have been proposed in clinical and experimental studies, such as: 1. Free radical scavengers (e.g., amifostine), mainly used to reduce DNA damage caused by photon radiotherapy, but with limited protective effects against high LET radiation; 2. DNA repair promoters, designed to enhance cellular self-repair capabilities, but often suffer from insufficient specificity or significant side effects. Therefore, existing protective measures are not ideal for cell damage caused by heavy ion radiation, especially in addressing the unique problem of the coexistence of necroptosis and inflammatory responses.

[0004] Therefore, developing a drug that can specifically alleviate heavy ion radiation damage, improve cell survival, and reduce inflammatory responses has become a pressing technical challenge. Summary of the Invention

[0005] The purpose of this invention is to provide a compound or drug that can alleviate or prevent cell damage caused by heavy ion radiation. This drug can improve cell clonal survival rate, inhibit necrosis and apoptosis, and weaken inflammatory responses, thereby improving the survival ability of normal cells after heavy ion irradiation. The invention employs the following technical solution:

[0006] A first aspect of the present invention provides a compound for mitigating or preventing cell damage caused by heavy ion radiation, said compound being Necrosulfonamide, the structure of which is shown in Formula I:

[0007]

[0008] A second aspect of the present invention provides a medicament for alleviating or preventing cell damage caused by heavy ion radiation, comprising the above-mentioned compound and a pharmaceutically acceptable carrier;

[0009] A third aspect of the present invention provides the use of the above-described compounds or drugs in the preparation of drugs for alleviating or preventing cell damage caused by heavy ion radiation;

[0010] Furthermore, the intended use is selected from any one or more of the following: relieving or preventing cell damage, tissue inflammatory response, and treatment-related adverse reactions caused during heavy ion radiotherapy.

[0011] Furthermore, the method for alleviating or preventing cell damage caused during heavy ion radiotherapy is selected from improving cell survival rate and / or inhibiting necrotizing apoptosis.

[0012] Furthermore, the relief or prevention of tissue inflammatory response is achieved by weakening inflammation-related transcriptional signals;

[0013] Furthermore, the indicated use is in combination with heavy ion radiotherapy to improve cell survival, inhibit necrotic apoptosis, and / or reduce inflammatory responses;

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] 1. Specific protective effect: Under carbon ion radiation, it can significantly improve the cell clonal formation rate and survival ability, while it does not show a significant protective effect under traditional photon radiation such as X-rays, showing the specific application value of heavy ion radiation.

[0016] 2. Improve cell death patterns: Drug treatment can effectively inhibit carbon ion-induced necrotizing apoptosis and promote the transformation of cells to milder death patterns such as apoptosis, thereby reducing tissue damage and inflammatory response.

[0017] 3. Inhibition of inflammatory transcriptional features: Transcriptomics and pathway enrichment analysis results showed that the drug could downregulate the activity of inflammation-related pathways (such as NF-κB signaling and cytokine-mediated pathways), significantly reduce the expression of pro-inflammatory factors such as IL1A, IL1B, CXCL1, CXCL2, and CXCL8, and help alleviate the inflammatory microenvironment after radiation.

[0018] 4. Broad application prospects: This drug composition can be used in the future to alleviate normal tissue damage caused by heavy ion radiotherapy, improve patient tolerance and treatment safety, and has potential application value in tumor radiotherapy, aerospace radiation protection, occupational radiation protection, etc., with high potential for clinical translation and industrial application. Attached Figure Description

[0019] Figure 1 The relative clonal survival rates of A549 and MDA-MB-231 cells after treatment with a heavy ion radiation-protective agent under different radiation conditions of X-ray and carbon ion irradiation.

[0020] Figure 2 This diagram illustrates the regulatory effects of anti-heavy-ion radiation cytoprotective agents on carbon ion irradiation-induced necrosis and apoptosis and related transcriptomic features. The diagram includes changes in the localization of necrosis-related proteins (A), Western blot analysis (B), transmission electron microscopy observation (C), GSVA analysis results (DE), GO enrichment analysis (F), and differential gene expression analysis (G). Detailed Implementation

[0021] Necrosulfonamide is a necrosis-apoptosis inhibitor with a molecular weight of 461.47 and a chemical formula of C1. 18 H 15 N5O6S2, CAS No.: 1360614-48-7. Necrosulfonamide blocks necrosome formation by selectively targeting (MLKL), and is effective in the IC50 of human HT-29. 50 The value was 124 nM. Necrosulfonamide significantly reduced BV6 / DAC-induced MV4-11 cell death by preventing the interaction between the MLKL-RIP1-RIP3 necrotizing body complex and its downstream effectors; specifically blocking RIP3 activation of downstream necrosis; and inhibiting MLKL-mediated necrosis by blocking the function of the N-terminal CC domain of MLKL. In the presence of caspase inhibitors, necrosulfonamide is also an effective inhibitor of the STS-induced necrosis pathway.

[0022] Necrosulfonamide inhibits downstream necrosis activated by RIP3 by blocking the function of the N-terminal CC region of MLKL and suppressing MLKL-mediated necrosis. Necrosulfonamide has no effect on TNF-α and Smac mimetic-induced apoptosis in Panc-1 cells that do not express RIP3, even at concentrations up to 5 μM. Necrosulfonamide effectively inhibits necrosis in human cells but has no such effect in mouse cells.

[0023] The present invention will now be described in further detail with reference to the accompanying drawings and specific embodiments.

[0024] Example 1: Cell clone viability detection

[0025] A549 and MDA-MB-231 cells undergoing exponential growth were divided into three groups: a mock control group, X-ray irradiation groups (2 Gy, 4 Gy, 6 Gy), and carbon ion irradiation groups (1 Gy, 2 Gy, 3 Gy). Each group was treated with or without a heavy ion radiation-protective agent, followed by irradiation. Relative clonal survival rates were calculated using a colony formation assay. Results are shown below. Figure 1 As shown: Under carbon ion irradiation, the drug treatment significantly improved cell viability: MDA-MB-231: C-1 Gy increased by +188.4% (P = 0.000138), C-2 Gy increased by +61% (P = 0.0495); A549: C-1 Gy increased by +54.8% (P = 0.0027), C-2 Gy increased by +17.3% (P = 0.195), C-3 Gy increased by +40% (P = 0.07); Under X-ray irradiation, the drug treatment did not show a protective effect, and under some conditions even reduced cell viability (e.g., X-2 Gy -42.6%, X-4 Gy -44.6%, X-6 Gy -32.2%).

[0026] The heavy ion radiation-resistant cell protectant provided by this invention can specifically improve the cell clone survival rate under carbon ion irradiation, while having no significant protective effect against X-ray radiation, suggesting its specific application value in heavy ion radiation.

[0027] Example 2: Regulation of carbon ion-induced necrosis and apoptosis and transcriptomic features by anti-heavy ion radiation cytoprotective agents

[0028] A549 and MDA-MB-231 cells in logarithmic growth phase were treated with different conditions: mock control; X-ray irradiation (2 Gy, 8 Gy); carbon ion irradiation (2 Gy); and carbon ion irradiation combined with a heavy ion radiation-protective agent. Cells were collected at 24 and 48 hours for protein detection, electron microscopy observation, and transcriptomics analysis.

[0029] Figure 2 This diagram illustrates the regulation of carbon ion-induced necrosis and apoptosis by anti-heavy ion radiation cell protectants and related pathways. Figure 2 A: Changes in the distribution of key necrosis- and apoptosis-related proteins in the cytoplasm and membrane components under different types and doses of radiation. The results showed that carbon ion irradiation induced their accumulation on the membrane, while drug treatment effectively inhibited this process. Figure 2 B: Western blot results showed that the expression of necrosis-apoptosis-related proteins increased and the apoptosis-related protein Caspase-3 was downregulated in the carbon ion irradiation group; after drug treatment, the expression of necrosis-apoptosis markers decreased and Caspase-3 was upregulated. Figure 2 C: Transmission electron microscopy results showed that carbon ion irradiation of cells exhibited typical necrotizing and apoptotic characteristics (cytoplasmic swelling, nuclear membrane rupture, and mitochondrial fragmentation), and this phenomenon was significantly reduced after drug treatment. Figure 2 D: GSVA analysis showed that drug treatment reduced the activity of the necroptosis pathway and enhanced the activity of apoptosis-related pathways. Figure 2 E: GSVA heatmap results showed that drug treatment significantly weakened the activity of inflammation-related pathways, including NF-κB signaling and cytokine-mediated signaling. Figure 2 F:GO biological process enrichment analysis showed that drug treatment significantly downregulated genes related to inflammation and cell survival. Figure 2 G: Transcriptome differential gene analysis showed that the drug could reverse carbon ion irradiation-induced differentially expressed genes, especially inhibiting the expression of pro-inflammatory factors such as IL1A, IL1B, CXCL1, CXCL2, and CXCL8.

[0030] In summary, this embodiment demonstrates that anti-heavy ion radiation cell protectants can improve cell survival after heavy ion radiation by inhibiting carbon ion-induced necrotizing apoptosis, reshaping cell death patterns, and weakening inflammatory transcriptional features.

[0031] Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art may make various modifications and alterations without departing from the spirit and scope of the present invention, and all such modifications and alterations shall be within the protection scope of the present invention.

Claims

1. A compound for alleviating cell damage caused by heavy ion radiation, characterized in that, The compound is Necrosulfonamide, and its structure is shown in Formula I:

2. A drug for alleviating cell damage caused by heavy ion radiation, characterized in that, It consists of the compound of claim 1 and a pharmaceutically acceptable carrier.

3. Use of the compound of claim 1 or the drug of claim 2 in the preparation of a drug for alleviating cell damage caused by heavy ion radiation.

4. The use according to claim 3, characterized in that, The use of the drug for alleviating cell damage caused by heavy ion radiation is selected from any one or more of the following: cell damage, tissue inflammatory response, and treatment-related adverse reactions caused during heavy ion radiotherapy.

5. The use according to claim 4, characterized in that, The method for alleviating cell damage caused during heavy ion radiotherapy is to improve cell survival rate and / or inhibit necrosis and apoptosis.

6. The use according to claim 4, characterized in that, The reduction of tissue inflammatory response is achieved by weakening inflammation-related transcriptional signals.

7. The use according to claim 3, characterized in that, The indicated use is in combination with heavy ion radiotherapy to improve cell survival, inhibit necrotic apoptosis, and / or reduce inflammatory responses.

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