Colorectal cancer radiotherapy sensitization medicine and application thereof

Abstract

The invention discloses a colorectal cancer radiotherapy sensitization medicine and application thereof, belongs to the field of biological medicine, and particularly relates to a colorectal cancer radiotherapy sensitization medicine which comprises active pharmaceutical ingredients and a solvent, the active pharmaceutical ingredients comprise doxorubicin and a nitrogen-containing compound, the nitrogen-containing compound comprises quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole, and the nitrogen-containing compound comprises quinoline-6-carbohydrazide and The colorectal cancer radiotherapy sensitization medicine disclosed by the invention focuses on a key driving gene FOXP4 for colorectal cancer radiotherapy resistance, and FOXP4 protein is degraded through a ubiquitin-proteasome way, so that not only can the sensitivity of tumor cells to radiotherapy be remarkably improved and tumor growth be inhibited, but also damage to normal tissues is reduced; moreover, the problem of drug resistance easily generated by a traditional method is effectively avoided, the research and development cost is greatly reduced, clinical transformation is accelerated, and the method has a good clinical application prospect.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and particularly to a radiosensitizing drug for colorectal cancer radiotherapy and its application. Background Art

[0002] Colorectal cancer is one of the main malignant tumors of the digestive tract. For patients with locally advanced colorectal cancer, preoperative neoadjuvant radiotherapy (or chemoradiotherapy) is routinely used clinically to reduce the tumor burden and recurrence risk. However, the current efficacy of neoadjuvant radiotherapy is limited, and only about 30% of patients show good sensitivity. The vast majority of patients have varying degrees of radiotherapy resistance, and the tumor fails to shrink significantly or even recurs, seriously affecting the prognosis of patients. Therefore, the development of radiosensitization strategies has become the key to improving the treatment effect and the quality of life of patients.

[0003] Currently, the technical research on radiosensitization of colorectal cancer radiotherapy mainly includes the following aspects: (1) Application of chemical sensitizers: Clinically, fluorouracil drugs (such as 5-FU or capecitabine) are mainly used as the standard concurrent chemoradiotherapy regimen. At the same time, it is found that the combination of drugs such as irinotecan can further enhance the tumor control effect. These drugs enhance the sensitivity of tumors to radiotherapy by interfering with the DNA synthesis and repair processes. (2) Research on natural compounds: Studies have shown that natural compounds such as curcumin have good radiosensitizing effects on CD133⁺ cancer stem cell-like cells in rectal cancer and can effectively reduce the tolerance of tumor cells to radiotherapy. (3) Development of microbial therapy: Emerging microbial therapies such as Roseburia intestinalis can significantly improve the sensitivity of colorectal cancer to radiotherapy and improve the tumor treatment effect by regulating the intestinal microecology. (4) Use of proteasome inhibitors: Proteasome inhibitors (especially 20S CP inhibitors) can improve the sensitivity of tumor cells to radiation damage by blocking the DNA double-strand break repair process in tumor cells. (5) Gene targeting strategies: It is found that DNA repair-related genes such as LIG4 play a key role in the radiosensitizing effect of the natural compound curcumin on rectal cancer cells, suggesting that gene-level intervention is expected to improve the radiosensitivity of tumors. (6) Application of nanomaterials: In recent years, emerging multifunctional nanomaterials, such as gold nanoparticles, improve the radiosensitivity by increasing the local radiation dose deposition or targeting the delivery of sensitizing drugs to the tumor microenvironment, providing new treatment options for clinical practice.

[0004] Despite the significant progress made in the above-mentioned radiotherapy sensitization strategies, there are still many deficiencies and limitations in practical applications: (1) Toxic and side effects of chemical sensitizers: Although commonly used chemical sensitizers (such as fluorouracil drugs) can improve radiotherapy sensitivity to a certain extent, the sensitization efficiency is limited, and long-term high-dose use can cause obvious toxic and side effects, such as severe gastrointestinal reactions, myelosuppression, etc. In addition, some patients are prone to develop drug resistance to these drugs, further limiting the therapeutic effect. (2) Clinical limitations of natural compounds: Although natural compounds such as curcumin show good radiotherapy sensitization effects in cell and animal models, they have problems such as low bioavailability and poor drug stability, which greatly limit their clinical promotion and application, and at the same time increase the preparation and delivery costs. (3) Challenges of microbial therapy: The therapy of using specific microorganisms to regulate the intestinal flora to sensitize radiotherapy is still in the initial stage of research, with complex mechanisms and being greatly affected by the dynamic changes of the intestinal microecology and individual differences. In addition, there are potential risks of immune rejection and infection, and the safety and stability of clinical application need to be further evaluated. (4) Nonspecific toxicity of proteasome inhibitors: Although proteasome inhibitors can significantly enhance the radiotherapy effect, they lack targeting, and their nonspecific effects lead to damage to normal tissues. Long-term use may accumulate to produce multi-organ toxicity, severely limiting their clinical applicability. (5) Bottlenecks of gene targeting strategies: The application of gene editing and gene regulation technologies in radiotherapy sensitization faces problems such as low targeting efficiency and high off-target effects, and gene therapy technologies are complex and costly, making it difficult to be widely promoted. At the same time, gene editing is also accompanied by ethical controversies and potential long-term risks, limiting its clinical application scope. (6) Limitations of nanomaterials: Although the nanomaterial sensitization strategy has broad prospects, its preparation process is complex and costly; some nanomaterials also have potential biological toxicity and environmental pollution risks. In addition, how to achieve efficient and safe in vivo targeted delivery is still one of the technical bottlenecks, severely restricting its clinical translation. These strategies have significant deficiencies in terms of sensitization efficiency, targeting, safety, and cost-effectiveness. Therefore, there is an urgent need to develop more precise, efficient, economical, and safe colorectal cancer radiotherapy sensitizing drugs. Summary of the Invention

[0005] The purpose of the present invention is to provide a colorectal cancer radiotherapy sensitizing drug and its application, to improve the sensitivity of tumor cells to radiotherapy and inhibit tumor growth.

[0006] The technical solution adopted by the present invention to achieve the above purpose is as follows: A colorectal cancer radiotherapy sensitizing drug, comprising a drug active ingredient and a solvent; the drug active ingredient contains doxorubicin; the dosage ratio of the drug active ingredient to the solvent is 1 μmol: 0.2 - 1 L.

[0007] The present invention focuses on the key driver gene FOXP4 for radiotherapy resistance in colorectal cancer. By using doxorubicin to degrade FOXP4 protein through the ubiquitin-proteasome pathway, it can not only significantly improve the sensitivity of tumor cells to radiotherapy, but also reduce the damage to normal tissues. Moreover, it effectively avoids the problem of drug resistance easily generated by traditional methods. In terms of clinical application, this scheme can be seamlessly connected with the existing radiotherapy scheme, and can achieve inhibition by using mature drugs such as doxorubicin, greatly reducing the R & D cost and accelerating clinical transformation, with good clinical application prospects.

[0008] Preferably, the solvent includes at least dimethyl sulfoxide.

[0009] Preferably, the drug active ingredient also contains doxorubicin and nitrogen-containing compounds. In the radiotherapy sensitizer for colorectal cancer of the present invention, nitrogen-containing compounds are also added. The nitrogen-containing compounds include quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole, which can not only bind to the macromolecular free radicals generated by radiation, fix the biomolecular damage caused by free radicals, and synergistically enhance the radiation sensitivity of tumor cells, significantly improving the radiation sensitization ratio; but also can inhibit tumor growth through the dual effects of targeting the FOXP4 gene and fixing the loss of free radicals, thereby optimizing the overall radiotherapy sensitization effect.

[0010] More preferably, the nitrogen-containing compounds include quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole.

[0011] Even more preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin.

[0012] Even more preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin.

[0013] Preferably, the radiotherapy sensitizer for colorectal cancer includes doxorubicin and dimethyl sulfoxide.

[0014] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol: 0.2-1 L.

[0015] Preferably, the radiotherapy sensitizer for colorectal cancer also includes doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and dimethyl sulfoxide.

[0016] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol: 0.2-1 L.

[0017] More preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin.

[0018] More preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin.

[0019] Preferably, the radiosensitizing drug for colorectal cancer radiotherapy further includes doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole, glycyrrhizoflavanone A, and dimethyl sulfoxide. In the present invention, glycyrrhizoflavanone A is further used in the radiosensitizing drug for colorectal cancer radiotherapy. By playing a role in hindering the proliferation of tumor cells and inducing DNA and mitochondrial damage in tumor cells, it can not only enhance the sensitivity of tumor cells to radiation, but also interfere with the energy metabolism and genetic material stability of tumor cells, producing a synergistic anti-tumor effect with radiotherapy, thereby significantly enhancing the radiosensitizing effect and tumor suppression effect of the overall treatment plan.

[0020] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol: 0.2 - 1 L.

[0021] More preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10 - 20% of the mass of doxorubicin.

[0022] More preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4 - 10% of the mass of doxorubicin.

[0023] More preferably, the mass of glycyrrhizoflavanone A is equivalent to 1 - 4% of the mass of doxorubicin.

[0024] The present invention also discloses the application of the radiosensitizing drug for colorectal cancer radiotherapy in the preparation of a drug for inhibiting the expression of the FOXP4 gene.

[0025] The present invention also discloses the application of the radiosensitizing drug for colorectal cancer radiotherapy in the preparation of an anti-tumor drug targeting the FOXP4 gene.

[0026] The present invention also discloses the application of the radiosensitizing drug for colorectal cancer radiotherapy in the preparation of a drug for promoting ferroptosis of tumor cells.

[0027] The present invention also discloses the application of the radiosensitizing drug for colorectal cancer radiotherapy in the preparation of a drug for inhibiting the growth of digestive system tumors.

[0028] Since the present invention uses doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole, glycyrrhizoflavanone A, and dimethyl sulfoxide to form the radiosensitizing drug for colorectal cancer radiotherapy, it has the following beneficial effects: The radiosensitizing drug for colorectal cancer radiotherapy provided by the present invention targets and inhibits the protein expression level of the FOXP4 gene, thereby activating the ferroptosis pathway to enhance radiation-induced cell killing; The radiosensitizing drug for colorectal cancer radiotherapy provided by the present invention is used in combination with radiotherapy, not only improving the synergistic radiotherapy sensitizing effect, with a radiation sensitization ratio of 1.62 - 1.79, but also significantly inhibiting tumor growth, and the tumor growth volume is reduced to 104 - 158 mm 3, showing significant clinical application prospects. Therefore, the present invention is a colorectal cancer radiotherapy sensitizing drug with excellent radiotherapy sensitizing effect and excellent tumor growth inhibition effect. Description of the Drawings

[0029] Figure 1 It is the expression of FOXP4 and GPX4 proteins.

[0030] Figure 2 It is the result of the HCT116 cell colony formation experiment.

[0031] Figure 3 It is the result of the HCT15 cell colony formation experiment.

[0032] Figure 4 It is the tumor growth curve.

[0033] Figure 5 It is the protein expression level of the FOXP4 gene after treatment with doxorubicin.

[0034] Figure 6 It is the sensitizing effect of doxorubicin.

[0035] Figure 7 It is the tumor suppression effect of doxorubicin combined with radiotherapy. Detailed Embodiments

[0036] The present invention will be further described in detail below in combination with the specific embodiments. The provided embodiments are only for clarifying the present invention, rather than limiting the scope of the present invention. The following provided embodiments can be used as a guide for those of ordinary skill in the art to make further improvements, and do not constitute any limitation to the present invention in any way.

[0037] The experimental methods in the following embodiments are all conventional methods unless otherwise specified. The materials, reagents, etc. used in the following embodiments can be obtained from commercial channels unless otherwise specified.

[0038] Example 1: A colorectal cancer radiotherapy sensitizing drug, including doxorubicin (Doxorubicin, DOX) and dimethyl sulfoxide. Doxorubicin was purchased from Shanghai TargetMol Biotechnology Co., Ltd., and the dosage ratio of doxorubicin to dimethyl sulfoxide was 1 μmol: 1 L.

[0039] Example 2: A colorectal cancer radiotherapy sensitizing drug, compared with Example 1, except that the dosage ratio of doxorubicin to dimethyl sulfoxide was changed to 1 μmol: 0.5 L.

[0040] Example 3: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that the dosage ratio of doxorubicin to dimethyl sulfoxide is changed to 1 μmol: 0.2 L.

[0041] Example 4: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole are additionally added, the mass of quinoline-6-carbohydrazide is equivalent to 20% of the mass of doxorubicin, and the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 10% of the mass of doxorubicin, and other conditions are the same as those in Example 1.

[0042] Example 5: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole are additionally added, the mass of quinoline-6-carbohydrazide is equivalent to 10% of the mass of doxorubicin, and the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 10% of the mass of doxorubicin, and other conditions are the same as those in Example 1.

[0043] Example 6: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole are additionally added, the mass of quinoline-6-carbohydrazide is equivalent to 20% of the mass of doxorubicin, and the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4% of the mass of doxorubicin, and other conditions are the same as those in Example 1.

[0044] Example 7: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and glycyrrhizin are additionally added, the mass of quinoline-6-carbohydrazide is equivalent to 20% of the mass of doxorubicin, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 10% of the mass of doxorubicin, and the mass of glycyrrhizin is equivalent to 4% of the mass of doxorubicin, and other conditions are the same as those in Example 1.

[0045] Example 8: A radiosensitizing drug for colorectal cancer. Compared with Example 1, except that quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and glycyrrhizin are additionally added, the mass of quinoline-6-carbohydrazide is equivalent to 20% of the mass of doxorubicin, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 10% of the mass of doxorubicin, and the mass of glycyrrhizin is equivalent to 1% of the mass of doxorubicin, and other conditions are the same as those in Example 1.

[0046] Comparative Example 1: A radiosensitizing drug for colorectal cancer radiotherapy. Compared with Example 1, except for the additional addition of quinoline-6-carbohydrazide, and the mass of quinoline-6-carbohydrazide being equivalent to 20% of the mass of doxorubicin, other conditions are the same as in Example 1.

[0047] Comparative Example 2: A radiosensitizing drug for colorectal cancer radiotherapy. Compared with Example 1, except for the additional addition of N-benzyloxymethyl-4-nitroimidazole, and the mass of N-benzyloxymethyl-4-nitroimidazole being equivalent to 10% of the mass of doxorubicin, other conditions are the same as in Example 1.

[0048] Comparative Example 3: A radiosensitizing drug for colorectal cancer radiotherapy. Compared with Example 1, except for the additional addition of licoricidin, and the mass of licoricidin being equivalent to 4% of the mass of doxorubicin, other conditions are the same as in Example 1.

[0049] Experimental Example: 1. Radiosensitizing effect of FOXP4 The expression of the FOXP4 gene was knocked down using lentivirus-mediated RNA interference technology. The FOXP4 interference sequence: 5’-TTCGCCTATTTCCGCAGAA-3’, and its nucleotide sequence is shown as SEQ ID No.1. This sequence was cloned into the pLKO.1-puro lentiviral vector and co-transfected with the packaging plasmids psPAX2 and pMD2.G into 293T cells for virus packaging. After collecting the virus supernatant, it was used to infect HCT116 or HCT15 colorectal cancer cells. After 48 hours of infection, puromycin at a concentration of 2 µg / mL was used for screening for 7 days to obtain a cell line stably expressing shFOXP4. The shFOXP4 cell lines were grouped into: HCT116 control group (HCT116-NC) cells, HCT116 knockdown group (HCT116-Sh) cells, HCT15 control group (HCT15-NC) cells, and HCT15 knockdown group (HCT15-Sh) cells. Among them, the HCT116-NC cells and HCT15-NC cells were not subjected to FOXP4 gene knockdown treatment.

[0050] The expressions of FOXP4, GPX4, and TUBULIN in HCT116-NC cells, HCT116-Sh cells, HCT15-NC cells, and HCT15-Sh cells were detected by Western blot.

[0051] Figure 1It is the expression of FOXP4 and GPX4 proteins. Compared with the control group, the protein expression levels of FOXP4 and GPX4 in HCT116-Sh cells and HCT15-Sh cells were significantly decreased, indicating that a FOXP4 gene knockdown model was successfully constructed in HCT116 and HCT15 cells, and inhibiting the expression of FOXP4 would reduce the protein expression level of GPX4. GPX4 is an antioxidant enzyme that can scavenge lipid peroxides, thereby inhibiting ferroptosis. Inhibiting FOXP4 expression would reduce the GPX4 expression level, leading to the accumulation of intracellular ROS and lipid peroxides, thereby restoring the activity of the ferroptosis pathway and enhancing the killing effect of radiotherapy on tumors.

[0052] Colony formation assays were performed on HCT116-NC cells, HCT116-Sh cells, HCT15-NC cells, and HCT15-Sh cells respectively. The specific steps are as follows: The cells were seeded in 6-well plates. After they adhered, they were respectively treated with X-rays at doses of 0, 2, 4, and 6 Gy, and three replicates were performed for each treatment. After irradiation, the cells were cultured for 9 - 12 days. After colony formation, they were fixed with methanol and stained with crystal violet. After colony formation, the colonies with ≥50 cells in each well were counted, the dose-survival curve was plotted, and the single-hit multi-target model was used for fitting. The fitting equation is: SF = 1 - (1 - e -kD ) n , where SF is the survival fraction, D is the irradiation dose, and k and n are the model fitting parameters. According to this model, the following parameters can be deduced: the mean lethal dose D0 = 1 / k, and the shoulder dose Dq = ln(n) × D0. The radiation sensitization ratio (SER) is calculated by comparing the shoulder doses of the cell survival curves before and after intervention. The formula is SER = Dq of the treatment group / Dq of the control group. SER > 1 indicates that the treatment group is more sensitive to radiotherapy and has a radiation sensitizing effect.

[0053] Figure 2 It is the result of the colony formation assay of HCT116 cells, Figure 3 It is the result of the colony formation assay of HCT15 cells. The results of the colony formation assay showed that the number of surviving colonies in the HCT116 cell line decreased significantly, and the calculated radiation sensitization ratio increased by about 1.5 times. The radiation sensitization ratio of the HCT15 cell line increased by about 1.13 times. This indicates that knocking down FOXP4 increases the sensitivity of colorectal cancer cells to radiation.

[0054] Four groups were set up in the mouse xenograft tumor model, namely the Sh-NC group, the Sh-FOXP4 group, the Sh-NC+IR group, and the Sh-FOXP4+IR group. In the Sh-NC group, HCT15-NC cells were subcutaneously inoculated into nude mice without X-ray irradiation treatment; in the Sh-FOXP4 group, HCT15-Sh cells stably transfected with shFOXP4 were subcutaneously inoculated into nude mice without X-ray irradiation treatment; in the Sh-NC+IR group, HCT15-NC cells were subcutaneously inoculated into nude mice, and after the tumor volume reached 100 mm 3 ³, 8 Gy×3 fractions of fractionated irradiation were given; in the Sh-FOXP4+IR group, HCT15-Sh cells stably transfected with shFOXP4 were subcutaneously inoculated into nude mice, and after the tumor volume reached 100 mm 3 ³, 8 Gy×3 fractions of fractionated irradiation were given. The tumor volume of each group was measured every 3 days since the treatment. The long diameter (L) and short diameter (W) were recorded using a vernier caliper, and the tumor volume was calculated according to the following formula: V = L×W 2 ² / 2.

[0055] Figure 4 Shown as the tumor growth curve. There was no significant difference in tumor volume between the Sh-NC group and the Sh-FOXP4 group, indicating that knocking down the expression level of the FOXP4 gene in the mouse xenograft tumor model could not effectively inhibit tumor growth. The tumor volumes of the Sh-NC+IR group and the Sh-FOXP4+IR group were significantly lower than those of the Sh-NC group and the Sh-FOXP4 group, and the tumor volume of the Sh-FOXP4+IR group was significantly reduced, indicating that knocking down FOXP4 combined with radiotherapy increased the tumor inhibitory effect.

[0056] It can be seen from Figures 1 to 4 that knocking down FOXP4 and combining it with radiotherapy can significantly improve the tumor inhibitory effect, and knocking down FOXP4 can make tumor cells regain sensitivity to radiation.

[0057] 2. Detection of drug inhibition of FOXP4 expression HCT116 cells were inoculated into 24-well plates at a density of 5×10 4 ⁵ cells / mL and divided into a control group and 3 experimental groups. The cells in the control group were treated with sterile distilled water for 24 h, and the 3 experimental groups were sequentially treated with the colorectal cancer radiotherapy sensitizing drugs of Examples 1-3 for 12 h. Then, the protein expression level of the FOXP4 gene in the control group and experimental group cells was detected by Western blot.

[0058] Figure 5The protein expression level of the FOXP4 gene after doxorubicin treatment. S1 represents the FOXP4 expression in the control group, S2 represents the FOXP4 expression in Example 1 group, S3 represents the FOXP4 expression in Example 2 group, and S4 represents the FOXP4 expression in Example 3 group. This indicates that doxorubicin, the active ingredient in the colorectal cancer radiotherapy sensitizing drug of the present invention, can significantly reduce the expression level of the FOXP4 gene in tumor cells, and the level of FOXP4 in tumor cells is reduced in a concentration-dependent manner by doxorubicin treatment, thereby activating the ferroptosis pathway and enhancing the cell killing induced by radiation.

[0059] 3. Detection of the sensitizing effect of the colorectal cancer radiotherapy sensitizing drug HCT15 cells were seeded in 6-well plates, and a control group and 9 experimental groups were set up. After they adhered, they were respectively treated with X-ray doses of 0, 2, 4, and 6 Gy, and each treatment was repeated three times. After irradiation, the cells were cultured for 9 - 12 days. After colony formation, they were fixed with methanol and stained with crystal violet. The control group was not treated with the colorectal cancer radiotherapy sensitizing drug, and the 9 experimental groups were successively treated with the colorectal cancer radiotherapy sensitizing drugs of Example 1, Examples 4 - 8, and Comparative Examples 1 - 3 for 12 h. After colony formation, the colonies with ≥50 cells in each well were counted, the dose-survival curve was plotted, and the single-hit multi-target model was used for fitting. The fitting equation is: SF = 1 - (1 - e-kD)n, where SF is the survival fraction, D is the irradiation dose, and k and n are the model fitting parameters. According to this model, the following parameters can be deduced: the mean lethal dose D0 = 1 / k, and the shoulder dose Dq = ln(n) × D0. The radiation sensitization ratio (SER) was calculated by comparing the shoulder doses of the cell survival curves before and after intervention. The formula is SER = Dq of the treatment group / Dq of the control group. SER > 1 indicates that the treatment group is more sensitive to radiotherapy and has a radiation sensitizing effect.

[0060] Table 1 Radiation sensitization ratio

[0061] Figure 6 This is the sensitizing effect of doxorubicin. The results showed that when the colorectal cancer radiotherapy sensitizing drug of Example 1 of the present invention was used in combination with radiotherapy to treat tumor cells, the survival fraction of tumor cells was significantly reduced, and the radiation sensitization ratio was increased to about 1.62 times. This indicates that the colorectal cancer radiotherapy sensitizing drug of the present invention shows a significant synergistic sensitizing effect when used in combination with radiotherapy, and significantly improves the radiotherapy sensitivity of tumor cells.

[0062] As can be seen from Table 1, the radiosensitization ratios of Examples 4-6 are higher than those of Example 1 because in the radiosensitizing drugs for colorectal cancer radiotherapy, quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside are additionally added in Examples 4-6, while quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside are not added in Example 1; the radiosensitization ratios of Examples 4-6 are higher than those of Comparative Example 1 and Comparative Example 2 because in the radiosensitizing drugs for colorectal cancer radiotherapy, only N-benzyloxymethyl-4-nitroimidazole glycoside is additionally added in Comparative Example 1, and only quinoline-6-carbohydrazide is additionally added in Comparative Example 2; the radiosensitization ratio of Example 4 is higher than those of Example 5 and Example 6 because in the radiosensitizing drugs for colorectal cancer radiotherapy, the usage amounts of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside are different. This shows that introducing quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside into the radiosensitizing drugs for colorectal cancer radiotherapy can effectively improve the radiosensitization ratio, and compared with using quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside alone, using quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside synergistically makes the sensitizing effect of the radiosensitizing drugs for colorectal cancer radiotherapy of the present invention better.

[0063] The radiosensitization ratios of Examples 7-8 are higher than those of Example 4 because in the radiosensitizing drugs for colorectal cancer radiotherapy, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole glycoside and glyasperin A are additionally added in Examples 7-8, while only quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside are additionally added in Example 4; the radiosensitization ratios of Examples 7-8 are higher than those of Comparative Example 3 because in the radiosensitizing drugs for colorectal cancer radiotherapy, only glyasperin A is additionally added in Comparative Example 3, while quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole glycoside are not added; the radiosensitization ratio of Example 7 is higher than that of Example 8 because in the radiosensitizing drugs for colorectal cancer radiotherapy, the usage amount of glyasperin A is different. This shows that further adding glyasperin A to the radiosensitizing drugs for colorectal cancer radiotherapy of the present invention helps to further improve the sensitizing effect of the radiosensitizing drugs for colorectal cancer radiotherapy.

[0064] 4. Detection of the tumor growth inhibitory effect of the radiosensitizing drugs for colorectal cancer radiotherapy The mouse xenograft tumor model was divided into a blank group, a control group and 9 experimental groups. The blank group was not treated with the radiosensitizing drugs for colorectal cancer radiotherapy and was treated with radiation; the control group was treated with the radiosensitizing drugs for colorectal cancer radiotherapy of Example 1 and was not treated with radiation; the 9 experimental groups were successively treated with the radiosensitizing drugs for colorectal cancer radiotherapy of Examples 1-6 and Comparative Examples 1-3 and were all treated with radiation. The tumor volume was recorded every three days for a total of 12 days. The tumor volume after 12 days of treatment.

[0065] Table 2 Tumor volume (mm 3 )

[0066] Figure 7 The tumor suppression effect of doxorubicin combined with radiotherapy. The tumor suppression effect of using the colorectal cancer radiotherapy sensitizing drug of Example 1 combined with radiation treatment is better than that of the blank group and the control group. This shows that compared with the single use of the colorectal cancer radiotherapy sensitizing drug or radiation treatment, the combination of the colorectal cancer radiotherapy sensitizing drug and radiation treatment can effectively inhibit tumor growth.

[0067] As can be seen from Table 2, the tumor volumes of Examples 4-6 are lower than those of Example 1 because in the colorectal cancer radiotherapy sensitizing drug, Examples 4-6 additionally added quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside, while Example 1 did not add quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the tumor volumes of Examples 4-6 are lower than those of Comparative Example 1 and Comparative Example 2 because in the colorectal cancer radiotherapy sensitizing drug, Comparative Example 1 only additionally added N-benzyloxymethyl-4-nitroimidazoside, and Comparative Example 2 only additionally added quinoline-6-carbohydrazide; the tumor volume of Example 4 is lower than those of Example 5 and Example 6 because in the colorectal cancer radiotherapy sensitizing drug, the usage amounts of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside are different. This shows that the introduction of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside into the colorectal cancer radiotherapy sensitizing drug can effectively inhibit tumor growth, and compared with the single use of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside, the synergistic use of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside makes the colorectal cancer radiotherapy sensitizing drug of the present invention have a better effect on inhibiting tumor growth.

[0068] The tumor volumes of Examples 7-8 are lower than those of Example 2 because in the colorectal cancer radiotherapy sensitizing drug, Examples 7-8 additionally added quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazoside and licoricidin, while Example 2 only additionally added quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the tumor volumes of Examples 7-8 are lower than those of Comparative Example 3 because in the colorectal cancer radiotherapy sensitizing drug, Comparative Example 3 only additionally added licoricidin and did not add quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the tumor volume of Example 7 is lower than that of Example 8 because in the colorectal cancer radiotherapy sensitizing drug, the usage amount of licoricidin is different. This shows that the further addition of licoricidin to the colorectal cancer radiotherapy sensitizing drug of the present invention helps to further improve the effect of the colorectal cancer radiotherapy sensitizing drug on inhibiting tumor growth.

[0069] The conventional operations in the operation steps of the present invention are well known to those skilled in the art and will not be elaborated here.

[0070] The above-described embodiments have elaborated on the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not used to limit the present invention. Any changes or modifications made within the scope of the principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A radiosensitizing drug for colorectal cancer radiotherapy, comprising a drug active ingredient and a solvent; the drug active ingredient contains doxorubicin; the dosage ratio of the drug active ingredient to the solvent is 1 μmol: 0.2 - 1 L.

2. The radiosensitizing drug for colorectal cancer radiotherapy according to claim 1, wherein The solvent at least includes dimethyl sulfoxide.

3. The radiosensitizing drug for colorectal cancer radiotherapy according to claim 1, wherein The drug active ingredient also contains doxorubicin and a nitrogen-containing compound.

4. The radiosensitizing drug for colorectal cancer radiotherapy according to claim 3, wherein The nitrogen-containing compound includes quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole.

5. The radiosensitizing drug for colorectal cancer radiotherapy according to claim 4, characterized in that, The mass of the quinoline-6-carbohydrazide is equivalent to 10 - 20% of the mass of doxorubicin.

6. The radiosensitizing drug for colorectal cancer radiotherapy according to claim 4, wherein The mass of the N-benzyloxymethyl-4-nitroimidazole is equivalent to 4 - 10% of the mass of doxorubicin.

7. Use of the radiosensitizing drug for colorectal cancer radiotherapy according to any one of claims 1 - 6 in the preparation of a drug for inhibiting the expression of the FOXP4 gene.

8. Use of the radiosensitizing drug for colorectal cancer radiotherapy according to any one of claims 1 - 6 in the preparation of an anti-tumor drug targeting the FOXP4 gene.

9. Use of the radiosensitizing drug for colorectal cancer radiotherapy according to any one of claims 1 - 6 in the preparation of a drug for promoting ferroptosis of tumor cells.

10. Use of the radiosensitizing drug for colorectal cancer radiotherapy according to any one of claims 1 - 6 in the preparation of a drug for inhibiting the growth of digestive system tumors.

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