Colorectal cancer radiotherapy sensitization drug and application thereof

Abstract

The present invention discloses a colorectal cancer radiotherapy sensitizing drug and its application, belonging to the field of biomedicine, and specifically relates to a colorectal cancer radiotherapy sensitizing drug, comprising a pharmaceutical active ingredient and a solvent, wherein the pharmaceutical active ingredient comprises doxorubicin and a nitrogen-containing compound, wherein the nitrogen-containing compound comprises quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole. The colorectal cancer radiotherapy sensitizing drug disclosed in the present invention focuses on FOXP4, a key driver gene of colorectal cancer radiotherapy resistance, and degrades FOXP4 protein through the ubiquitin-proteasome pathway. This not only significantly increases the sensitivity of tumor cells to radiotherapy, inhibits tumor growth, and reduces damage to normal tissues; it also effectively circumvents the drug resistance problem that is prone to occur with traditional methods, significantly reduces R&D costs, accelerates clinical transformation, and has good clinical application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a colorectal cancer radiotherapy sensitization drug and application thereof. Background Art

[0002] Colorectal cancer is one of the major gastrointestinal malignancies. For patients with locally advanced colorectal cancer, preoperative neoadjuvant radiotherapy (or chemoradiotherapy) is routinely used clinically to reduce tumor burden and the risk of recurrence. However, the efficacy of neoadjuvant radiotherapy is currently limited, with only approximately 30% of patients demonstrating good sensitivity. The vast majority of patients exhibit varying degrees of radioresistance, resulting in failure to significantly reduce tumor size or even recurrence, seriously impacting patient prognosis. Therefore, the development of radiosensitization strategies is crucial to improving treatment efficacy and patient quality of life.

[0003] At present, the technical research on radiosensitization of colorectal cancer mainly includes the following aspects: (1) Application of chemosensitizers: In clinical practice, fluorouracil drugs (such as 5-FU or capecitabine) are mainly used as standard synchronous chemoradiotherapy regimens. At the same time, studies have found that combined use of drugs such as irinotecan can further enhance the tumor control effect. These drugs enhance the sensitivity of tumors to radiotherapy by interfering with DNA synthesis and repair processes. (2) Research on natural compounds: Studies have shown that natural compounds such as curcumin have a good radiosensitization effect on CD133⁺ tumor stem-like cells of 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 increase the sensitivity of colorectal cancer to radiotherapy and improve the treatment effect of tumors by regulating the intestinal microecology. (4) Use of proteasome inhibitors: Proteasome inhibitors (especially 20S CP inhibitors) can increase the sensitivity of tumor cells to radiation damage by blocking the DNA double-strand break repair process of tumor cells. (5) Gene targeting strategy: Studies have found that DNA repair-related genes such as LIG4 play a key role in the radiosensitization effect of the natural compound curcumin on rectal cancer cells, suggesting that genetic intervention is expected to improve tumor radiosensitivity. (6) Application of nanomaterials: Multifunctional nanomaterials, such as gold nanoparticles, have emerged in recent years. By increasing local radiation dose deposition or targeted delivery of sensitizing drugs to the tumor microenvironment, they can improve radiosensitivity and provide new treatment options for the clinic.

[0004] Although the above-mentioned radiosensitization strategies have made significant progress, there are still many deficiencies and limitations in practical applications: (1) Toxic side effects of chemosensitizers: Although commonly used chemosensitizers (such as fluorouracil drugs) can improve radiosensitivity to a certain extent, the sensitization efficiency is limited, and long-term high-dose use can lead to significant toxic side effects, such as severe gastrointestinal reactions and bone marrow suppression. In addition, some patients are prone to develop resistance to these drugs, further limiting the efficacy. (2) Clinical limitations of natural compounds: Although natural compounds such as curcumin have shown good radiosensitization 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 increase the cost of preparation and delivery. (3) Challenges of microbial therapy: The use of specific microorganisms to regulate intestinal flora to enhance radiosensitivity is still in the early stages of research. Its mechanism is complex and is greatly affected by the dynamic changes of intestinal microecology and individual differences. In addition, there is a potential risk 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 effect of radiotherapy, they lack targeting and their nonspecific effects can lead to normal tissue damage. Long-term use may result in cumulative multi-organ toxicity, which severely limits their clinical applicability. (5) Bottlenecks of gene targeting strategies: The application of gene editing and gene regulation technologies in radiotherapy sensitization faces the problems of low targeting efficiency and high off-target effects. In addition, gene therapy technology is complex and expensive, making it difficult to promote it widely. At the same time, gene editing is also accompanied by ethical controversies and potential long-term risks, which limits its clinical application scope. (6) Limitations of nanomaterials: Although nanomaterial sensitization strategies have broad prospects, their preparation processes are complex and expensive; some nanomaterials also have potential biological toxicity and environmental pollution risks. In addition, how to achieve efficient and safe in vivo targeted delivery remains one of the technical bottlenecks, which seriously restricts its clinical transformation. These strategies have significant defects in sensitization efficiency, targeting, safety and cost-effectiveness. Therefore, there is an urgent need to develop more accurate, efficient, economical and safe colorectal cancer radiosensitization drugs. Summary of the Invention

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

[0006] The technical solutions adopted by the present invention to achieve the above-mentioned purpose are:

[0007] A colorectal cancer radiotherapy sensitization drug comprises a pharmaceutical active ingredient and a solvent; the pharmaceutical active ingredient contains doxorubicin; and the dosage ratio of the pharmaceutical active ingredient to the solvent is 1 μmol:0.2-1L.

[0008] This approach targets FOXP4, a key driver of radiotherapy resistance in colorectal cancer. Using doxorubicin to degrade FOXP4 protein via the ubiquitin-proteasome pathway, it significantly increases the sensitivity of tumor cells to radiotherapy while minimizing damage to normal tissues. It also effectively circumvents the drug resistance that is common with traditional approaches. In terms of clinical application, this approach seamlessly integrates with existing radiotherapy regimens and utilizes established drugs like doxorubicin for inhibition, significantly reducing R&D costs and accelerating clinical translation, promising promising clinical application prospects.

[0009] Preferably, the solvent comprises at least dimethyl sulfoxide.

[0010] Preferably, the active ingredients of the drug also contain doxorubicin and a nitrogen-containing compound. The present invention also adds nitrogen-containing compounds to the colorectal cancer radiosensitization drug. The nitrogen-containing compounds include quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole. These compounds can not only bind to macromolecular free radicals generated by radiation, fix the biomolecular damage caused by free radicals, synergistically enhance the radiation sensitivity of tumor cells, and significantly improve the radiosensitization ratio; they can also inhibit tumor growth through the dual effects of targeting the FOXP4 gene and fixing the loss of free radicals, thereby optimizing the overall radiosensitization effect.

[0011] More preferably, the nitrogen-containing compound includes quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole.

[0012] More preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin.

[0013] More preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin.

[0014] Preferably, the colorectal cancer radiosensitization drugs include doxorubicin and dimethyl sulfoxide.

[0015] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol:0.2-1L.

[0016] Preferably, the colorectal cancer radiosensitization drug further comprises doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and dimethyl sulfoxide.

[0017] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol:0.2-1L.

[0018] More preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin.

[0019] More preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin.

[0020] Preferably, the colorectal cancer radiosensitization drug also includes doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole, glycyrrhizic acid isoflavanone, and dimethyl sulfoxide. The present invention further uses glycyrrhizic acid isoflavanone in the colorectal cancer radiosensitization drug. By inhibiting tumor cell proliferation 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 improving the sensitization effect and tumor inhibition effect of the overall treatment plan.

[0021] More preferably, the dosage ratio of doxorubicin to dimethyl sulfoxide is 1 μmol:0.2-1L.

[0022] More preferably, the mass of quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin.

[0023] More preferably, the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin.

[0024] More preferably, the mass of glycyrrhizin isoflavane is equivalent to 1-4% of the mass of doxorubicin.

[0025] The present invention also discloses the use of the colorectal cancer radiosensitization drug in the preparation of a drug for inhibiting FOXP4 gene expression.

[0026] The present invention also discloses the use of the colorectal cancer radiotherapy sensitization drug in the preparation of an anti-tumor drug targeting the FOXP4 gene.

[0027] The present invention also discloses the use of a colorectal cancer radiosensitization drug in the preparation of a drug for promoting ferroptosis of tumor cells.

[0028] The present invention also discloses the use of the colorectal cancer radiotherapy sensitization drug in the preparation of a drug for inhibiting the growth of digestive system tumors.

[0029] The present invention uses doxorubicin, quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole, glycyrrhizic acid isoflavone and dimethyl sulfoxide to form a colorectal cancer radiosensitization drug, so it has the following beneficial effects: the colorectal cancer radiosensitization drug 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 colorectal cancer radiosensitization drug provided by the present invention is used in combination with radiotherapy, not only improving the synergistic radiosensitization effect, with a radiosensitization ratio of 1.62-1.79, but also significantly inhibiting tumor growth, with the tumor growth volume reduced to 104-158 mm 3, showing significant clinical application prospects. Therefore, the present invention is a colorectal cancer radiosensitization drug with excellent radiosensitization effect and tumor growth inhibition effect. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The expression of FOXP4 and GPX4 proteins.

[0031] Figure 2 These are the results of HCT116 cell clone formation experiments.

[0032] Figure 3 These are the results of HCT15 cell clone formation experiments.

[0033] Figure 4 Tumor growth curve.

[0034] Figure 5 is the protein expression level of FOXP4 gene after doxorubicin treatment.

[0035] Figure 6 It is the sensitizing effect of doxorubicin.

[0036] Figure 7 The tumor inhibitory effect of doxorubicin combined with radiotherapy. DETAILED DESCRIPTION

[0037] The present invention will be further described in detail below in conjunction with specific embodiments. The examples provided are only for illustrating the present invention and are not intended to limit the scope of the present invention. The examples provided below can serve as a guide for further improvements by those skilled in the art and are not intended to limit the present invention in any way.

[0038] The experimental methods in the following examples are conventional methods unless otherwise specified. The materials and reagents used in the following examples are commercially available unless otherwise specified.

[0039] Example 1:

[0040] A colorectal cancer radiosensitizer comprises doxorubicin (DOX) and dimethyl sulfoxide. Doxorubicin was purchased from Shanghai Taoshu Biotechnology Co., Ltd., and the dosage ratio of doxorubicin to dimethyl sulfoxide was 1 μmol:1 L.

[0041] Example 2:

[0042] A colorectal cancer radiosensitization drug, compared with Example 1, except that the dosage ratio of doxorubicin and dimethyl sulfoxide is changed to 1 μmol:0.5L.

[0043] Example 3:

[0044] A colorectal cancer radiosensitization drug, compared with Example 1, except that the dosage ratio of doxorubicin and dimethyl sulfoxide is changed to 1 μmol:0.2L.

[0045] Example 4:

[0046] A colorectal cancer radiosensitization drug, 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, other conditions are the same as Example 1.

[0047] Example 5:

[0048] A colorectal cancer radiosensitization drug, 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, other conditions are the same as Example 1.

[0049] Example 6:

[0050] A colorectal cancer radiosensitization drug, 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, other conditions are the same as Example 1.

[0051] Example 7:

[0052] A colorectal cancer radiosensitization drug, compared with Example 1, except that quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and glycyrrhizic acid isoflavanone 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 glycyrrhizic acid isoflavanone is equivalent to 4% of the mass of doxorubicin, other conditions are the same as Example 1.

[0053] Example 8:

[0054] A colorectal cancer radiosensitization drug, compared with Example 1, except that quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazole and glycyrrhizic acid isoflavanone 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 glycyrrhizic acid isoflavanone is equivalent to 1% of the mass of doxorubicin, other conditions are the same as Example 1.

[0055] Comparative Example 1:

[0056] A colorectal cancer radiosensitization drug, compared with Example 1, except that quinoline-6-carbohydrazide is additionally added, and the mass of quinoline-6-carbohydrazide is equivalent to 20% of the mass of doxorubicin, other conditions are the same as Example 1.

[0057] Comparative Example 2:

[0058] A colorectal cancer radiosensitization drug, compared with Example 1, except that N-benzyloxymethyl-4-nitroimidazole is additionally added, and the mass of N-benzyloxymethyl-4-nitroimidazole is equivalent to 10% of the mass of doxorubicin, other conditions are the same as Example 1.

[0059] Comparative Example 3:

[0060] A colorectal cancer radiosensitization drug, compared with Example 1, except that glycyrrhizin isoflavanone is additionally added, and the mass of glycyrrhizin isoflavanone is equivalent to 4% of the mass of doxorubicin, other conditions are the same as Example 1.

[0061] Experimental example:

[0062] 1. Radiosensitization effect of FOXP4

[0063] Lentiviral-mediated RNA interference (RNAi) was used to knock down FOXP4 gene expression. The FOXP4 interference sequence (5'-TTCGCCTATTTCCGCAGAA-3') is shown in 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 viral packaging. Viral supernatants were collected and used to infect HCT116 or HCT15 colorectal cancer cells. Forty-eight hours after infection, cell lines stably expressing shFOXP4 were selected with puromycin at a concentration of 2 µg / mL for 7 days. The shFOXP4 cell lines were grouped into the following groups: HCT116 control (HCT116-NC), HCT116 knockdown (HCT116-Sh), HCT15 control (HCT15-NC), and HCT15 knockdown (HCT15-Sh). Among them, HCT116-NC cells and HCT15-NC cells were not treated with FOXP4 gene knockdown.

[0064] Western blot was used to detect the expression of FOXP4, GPX4 and TUBULIN in HCT116-NC cells, HCT116-Sh cells, HCT15-NC cells and HCT15-Sh cells.

[0065] Figure 1Figure 2 shows the protein expression of FOXP4 and GPX4. Compared to the control group, the protein expression levels of FOXP4 and GPX4 were significantly reduced in HCT116-Sh and HCT15-Sh cells. This indicates that a FOXP4 knockdown model was successfully established in HCT116 and HCT15 cells, and that inhibiting FOXP4 expression reduced GPX4 protein expression. GPX4 is an antioxidant enzyme that scavenges lipid peroxides, thereby inhibiting ferroptosis. Inhibiting FOXP4 expression reduces GPX4 expression, leading to the accumulation of intracellular ROS and lipid peroxides, thereby restoring the activity of the ferroptosis pathway and enhancing the tumor-killing effect of radiotherapy.

[0066] HCT116-NC cells, HCT116-Sh cells, HCT15-NC cells, and HCT15-Sh cells were subjected to a colony formation experiment. The specific steps were as follows: cells were seeded in 6-well plates and, after attachment, were treated with X-rays at doses of 0, 2, 4, and 6 Gy, respectively. Each treatment was repeated in triplicate. After irradiation, the cells were cultured for 9-12 days. After colonies formed, the cells were fixed with methanol and stained with crystal violet. After colony formation, colonies with ≥50 cells in each well were counted, and dose-survival curves were plotted. The single-shot multi-target model was used to fit the curves. The fitting equation was: SF = 1-(1-e -kD ) n , where SF is the survival fraction, D is the irradiation dose, and k and n are model fitting parameters. Based on this model, the following parameters can be derived: mean lethal dose D0 = 1 / k, shoulder dose Dq = ln(n) × D0. The radiosensitization ratio (SER) is calculated by comparing the shoulder doses of the cell survival curves before and after intervention using the formula SER = Dq of the treatment group / Dq of the control group. A SER > 1 indicates that the treatment group is more sensitive to radiotherapy, demonstrating a radiosensitizing effect.

[0067] Figure 2 The results of the HCT116 cell clone formation experiment are shown in Figure 2. Figure 3 Results from a HCT15 cell clonogenic assay showed a significant decrease in the number of surviving colonies in the HCT116 cell line, with a calculated radiosensitivity increase of approximately 1.5-fold and 1.13-fold, respectively. This suggests that knocking down FOXP4 increases the sensitivity of colorectal cancer cells to radiation.

[0068] Four groups were set up in the mouse xenograft tumor model: Sh-NC group, Sh-FOXP4 group, Sh-NC+IR group, and Sh-FOXP4+IR group. In the Sh-NC group, HCT15-NC cells were subcutaneously inoculated into nude mice without X-ray irradiation; in the Sh-FOXP4 group, HCT15-Sh cells stably transfected with shFOXP4 were subcutaneously inoculated into nude mice without X-ray irradiation; in the Sh-NC+IR group, HCT15-NC cells were subcutaneously inoculated into nude mice and the tumor volume reached 100 mm. 3 Then, 8 Gy × 3 fractions of irradiation were given; in the Sh-FOXP4+IR group, HCT15-Sh cells stably transfected with shFOXP4 were subcutaneously inoculated into nude mice, and the mice were treated with irradiation after the tumor volume reached 100 mm. 3 Then, 8 Gy × 3 fractions were given. The tumor volume of each group was measured every 3 days from the start of 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.

[0069] Figure 4 Figure 2 shows the tumor growth curve. There was no significant difference in tumor volume between the Sh-NC and Sh-FOXP4 groups, indicating that knocking down FOXP4 expression in mouse xenograft models does not effectively inhibit tumor growth. Tumor volumes in the Sh-NC+IR and Sh-FOXP4+IR groups were significantly lower than those in the Sh-NC and Sh-FOXP4 groups, with the Sh-FOXP4+IR group showing a significant decrease in tumor volume. This suggests that FOXP4 knockdown combined with radiotherapy enhances tumor suppression.

[0070] Depend on Figures 1 to 4 It can be seen that knocking down FOXP4 and combining it with radiotherapy can significantly improve the tumor suppression effect, and knocking down FOXP4 can make tumor cells regain sensitivity to radiation.

[0071] 2. Detection of drug inhibition of FOXP4 expression

[0072] HCT116 cells were cultured at 5 × 10 4 Cells were seeded at a density of 100 μg / ml in 24-well plates and divided into a control group and three experimental groups. The control group cells were treated with sterile distilled water for 24 hours, while the three experimental groups were treated with the colorectal cancer radiosensitization drugs described in Examples 1-3 for 12 hours, respectively. Western blot analysis was then used to determine the protein expression levels of the FOXP4 gene in the control and experimental groups.

[0073] Figure 5is the protein expression level of the FOXP4 gene after doxorubicin treatment, S1 is the FOXP4 expression of the control group, S2 is the FOXP4 expression of the Example 1 group, S3 is the FOXP4 expression of the Example 2 group, and S4 is the FOXP4 expression of the Example 3 group. This shows that doxorubicin, the active ingredient in the colorectal cancer radiosensitization drug of the present invention, can significantly reduce the expression level of the FOXP4 gene in tumor cells, and doxorubicin treatment reduces the level of FOXP4 in tumor cells in a concentration-dependent manner, thereby activating the ferroptosis pathway to enhance radiation-induced cell killing.

[0074] 3. Detection of the sensitizing effect of radiosensitizing drugs for colorectal cancer

[0075] HCT15 cells were seeded in 6-well plates. A control group and nine experimental groups were set up. After attachment, cells were treated with X-rays at doses of 0, 2, 4, and 6 Gy, respectively, with triplicates for each treatment. Following irradiation, cells were cultured for 9-12 days. After colony formation, cells were fixed with methanol and stained with crystal violet. The control group was not treated with colorectal cancer radiosensitizing drugs. The nine experimental groups were treated with the colorectal cancer radiosensitizing drugs described in Example 1, Examples 4-8, and Comparative Examples 1-3, respectively, for 12 hours. After colony formation, colonies with ≥50 cells per well were counted, and dose-survival curves were plotted. A single-target multi-target model was used for fitting, using the equation: SF = 1-(1-e-kD)n, where SF is the survival fraction, D is the irradiation dose, and k and n are model fitting parameters. Based on this model, the following parameters can be derived: mean lethal dose D0 = 1 / k, shoulder dose Dq = ln(n) × D0. The radiosensitization ratio (SER) is calculated by comparing the shoulder dose of the cell survival curve before and after the intervention using the formula: SER = Dq of the treatment group / Dq of the control group. A SER > 1 indicates that the treatment group is more sensitive to radiotherapy, demonstrating a radiosensitizing effect.

[0076] Table 1 Radiosensitization ratio

[0077]

[0078] Figure 6 The results showed that when the colorectal cancer radiosensitization 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 radiosensitization ratio was increased to approximately 1.62 times. This shows that the colorectal cancer radiosensitization drug of the present invention, when used in combination with radiotherapy, exhibits a significant synergistic sensitization effect, significantly improving the radiotherapy sensitivity of tumor cells.

[0079] As can be seen from Table 1, the radiosensitization ratio of Example 4-6 is higher than that of Example 1 because, in the colorectal cancer radiotherapy sensitizing drug, Example 4-6 additionally adds quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside, while Example 1 does not add quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the radiosensitization ratio of Example 4-6 is higher than that of Comparative Example 1 and Comparative Example 2 because, in the colorectal cancer radiotherapy sensitizing drug, Comparative Example 1 only additionally adds N-benzyloxymethyl-4-nitroimidazoside, while Comparative Example 2 only additionally adds quinoline-6-carbohydrazide; the radiosensitization ratio of Example 4 is higher than that of Example 5 and Example 6 because, in the colorectal cancer radiotherapy sensitizing drug, the amounts of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside used 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 improve the radiation sensitization ratio, and compared with the use of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside alone, 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 sensitization effect.

[0080] The radiosensitization ratio of Examples 7-8 is higher than that of Example 4 because, in the colorectal cancer radiosensitizing drug, Examples 7-8 additionally added quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazoside, and glycyrrhizic acid isoflavan methyl, while Example 4 only added quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the radiosensitization ratio of Examples 7-8 is higher than that of Comparative Example 3 because, in the colorectal cancer radiosensitizing drug, Comparative Example 3 only added glycyrrhizic acid isoflavan methyl, but did not add quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the radiosensitization ratio of Example 7 is higher than that of Example 8 because the amount of glycyrrhizic acid isoflavan methyl used in the colorectal cancer radiosensitizing drug is different. This shows that the further addition of glycyrrhizic acid isoflavan methyl to the colorectal cancer radiosensitizing drug of the present invention helps to further improve the sensitization effect of the colorectal cancer radiosensitizing drug.

[0081] 4. Detection of the tumor growth inhibition effect of colorectal cancer radiosensitization drugs

[0082] Mouse transplant tumor models were divided into a blank group, a control group, and nine experimental groups. The blank group was not treated with a colorectal cancer radiosensitizer but was treated with radiation; the control group was treated with the colorectal cancer radiosensitizer of Example 1 but was not treated with radiation; the nine experimental groups were treated with the colorectal cancer radiosensitizers of Examples 1-6 and Comparative Examples 1-3, respectively, and all were treated with radiation. Tumor volume was recorded every three days for a total of 12 days. Tumor volume after the 12th day of treatment was recorded.

[0083] Table 2 Tumor volume (mm3 )

[0084]

[0085] Figure 7 The tumor inhibition effect of doxorubicin combined with radiotherapy is shown in Figure 1. The tumor inhibition effect of the colorectal cancer radiosensitizing drug combined with radiation treatment is better than that of the blank group and the control group, which shows that the colorectal cancer radiosensitizing drug combined with radiation treatment can effectively inhibit tumor growth compared with the use of colorectal cancer radiosensitizing drug or radiation treatment alone.

[0086] As can be seen from Table 2, the tumor volume of Example 4-6 is lower than that of Example 1 because, in the colorectal cancer radiotherapy sensitizing drug, Example 4-6 additionally adds quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside, while Example 1 does not add quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the tumor volume of Example 4-6 is lower than that of Comparative Example 1 and Comparative Example 2 because, in the colorectal cancer radiotherapy sensitizing drug, Comparative Example 1 only additionally adds N-benzyloxymethyl-4-nitroimidazoside, while Comparative Example 2 only additionally adds quinoline-6-carbohydrazide; the tumor volume of Example 4 is lower than that of Example 5 and Example 6 because, in the colorectal cancer radiotherapy sensitizing drug, the amounts of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside used are different. This shows that the introduction of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside into the colorectal cancer radiotherapy sensitization drug can effectively inhibit tumor growth, and compared with the use of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside alone, the synergistic use of quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside makes the colorectal cancer radiotherapy sensitization drug of the present invention more effective in inhibiting tumor growth.

[0087] The tumor volume of Example 7-8 is lower than that of Example 2 because, in the colorectal cancer radiosensitizing drug, Examples 7-8 additionally added quinoline-6-carbohydrazide, N-benzyloxymethyl-4-nitroimidazoside, and glycyrrhizic acid isoflavan methyl, while Example 2 only added quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazoside; the tumor volume of Example 7-8 is lower than that of Comparative Example 3 because, in the colorectal cancer radiosensitizing drug, Comparative Example 3 only added glycyrrhizic acid isoflavan methyl, but 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 the amount of glycyrrhizic acid isoflavan methyl used in the colorectal cancer radiosensitizing drug is different. This shows that the further addition of glycyrrhizic acid isoflavan methyl to the colorectal cancer radiosensitizing drug of the present invention helps to further enhance the effect of the colorectal cancer radiosensitizing drug in inhibiting tumor growth.

[0088] The conventional operations in the operating steps of the present invention are well known to those skilled in the art and will not be described in detail here.

[0089] The embodiments described above provide a detailed description of 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 intended to limit the present invention. Any changes or modifications made within the scope of the principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A colorectal cancer radiosensitization drug, comprising a pharmaceutically active ingredient and a solvent; the pharmaceutically active ingredient comprises doxorubicin, a nitrogen-containing compound, and glycyrrhizic acid isoflavan methyl; the dosage ratio of the pharmaceutically active ingredient to the solvent is 1 μmol:0.2-1 L; the nitrogen-containing compound comprises quinoline-6-carbohydrazide and N-benzyloxymethyl-4-nitroimidazole; the mass of the quinoline-6-carbohydrazide is equivalent to 10-20% of the mass of doxorubicin; the mass of the N-benzyloxymethyl-4-nitroimidazole is equivalent to 4-10% of the mass of doxorubicin; and the mass of the glycyrrhizic acid isoflavan methyl is equivalent to 1-4% of the mass of doxorubicin.

2. A colorectal cancer radiosensitization drug according to claim 1, characterized in that: The solvent includes at least dimethyl sulfoxide.

3. Use of the colorectal cancer radiosensitization drug according to claim 1 or 2 in the preparation of a drug for inhibiting colorectal cancer tumor growth, characterized in that: The drug can target and inhibit FOXP4 gene expression and promote ferroptosis of tumor cells.

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