阅读说明：本技术 西达本胺及其衍生物的用途 (Application of xidabenamine and derivatives thereof ) 是由 黄旭 鲁先平 潘德思 王绿化 于 2020-10-29 设计创作，主要内容包括：本发明涉及药物技术领域,公开了西达本胺及其衍生物的用途。本发明揭示了西达本胺具有抗肿瘤放射治疗增敏作用,西达本胺联合放射治疗可以更强地抑制肿瘤细胞增殖和诱导细胞凋亡,同时抑制肿瘤细胞的干细胞特征,两者联合在动物体内表现出相比单独治疗更强的肿瘤抑制效果。西达本胺不仅可以增强放射治疗的抑瘤活性,同时可以降低肿瘤产生治疗抵抗和复发的风险,西达本胺联合放射治疗可以提供比单一治疗更优的持续疗效。(The invention relates to the technical field of medicines, and discloses application of cidentamine and derivatives thereof. The invention discloses that the cydariamine has the sensitization effect of anti-tumor radiotherapy, the combination of the cydariamine and the radiotherapy can more strongly inhibit the proliferation and the apoptosis of tumor cells and simultaneously inhibit the stem cell characteristics of the tumor cells, and the combination of the cydariamine and the radiotherapy shows stronger tumor inhibition effect in animals compared with the single treatment. The cydarifamid not only can enhance the tumor inhibition activity of radiotherapy, but also can reduce the treatment resistance and recurrence risk of tumors, and the combination of the cydarifamid and the radiotherapy can provide better continuous curative effect than single treatment.)

1. The application of the xidapamide and the derivatives thereof in preparing a radiation sensitizer; or the application of the combination of the Xidabenamine and the derivatives thereof with radiotherapy in the preparation of products for treating tumors; or the application of the cidentamine and the derivatives thereof in treating tumors in combination with radiotherapy.

2. The use according to claim 1, wherein the cidentamine and its derivatives comprise cidentamine and its amorphous forms, crystal forms, stereoisomers, geometric isomers, tautomers, solvates, metabolites, pharmaceutically acceptable salts, prodrugs and compositions comprising same.

3. The use of claim 1, wherein the tumor is selected from one or more of lung cancer, liver cancer, stomach cancer, breast cancer, ovarian cancer, lymphoma, leukemia, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, esophageal cancer, uterine cancer, vaginal cancer, prostate cancer, melanoma, urothelial cancer, and sarcoma.

4. Use according to claim 1, wherein the radiotherapy is a radiotherapy for the treatment of tumors.

5. A method of treating tumours, characterised in that the combination of cidentamine and derivatives thereof with radiotherapy.

6. The method of claim 5, wherein said radiation therapy is administered simultaneously or sequentially in any order with the administration of sildenamine and its derivatives.

Technical Field

The invention relates to the technical field of medicines, in particular to application of cidentamine and derivatives thereof.

Background

Radiotherapy is a common method for clinical tumor therapy, and is mainly a method of local treatment of tumors by using radioactive rays (such as alpha, beta, gamma rays generated by radioactive isotopes). The basic principle of radiotherapy is that DNA damage is caused by rays, and for tumor cells with active proliferation, the DNA damage repair cannot be effectively completed, so that damage accumulation is caused, cell apoptosis is induced, and the anti-tumor effect is achieved. The efficacy of radiation therapy depends on the sensitivity of the tumor to radiation and its resulting damage, and different tumor types and pathological stages affect the ultimate efficacy of radiation therapy. Meanwhile, the tumor can avoid the apoptosis induced by radiotherapy through different mechanisms to cause treatment resistance, so that the continuous curative effect of the radiotherapy is reduced. Increasing the dose of radiotherapy can promote the curative effect, but brings more side effects, so it is important how to better realize the curative effect safety of radiotherapy in clinic, and besides more precise directional treatment, it is also a potential development direction to adopt other treatment means to increase the sensitivity of the tumor to the radiotherapy or reduce the resistance of the tumor.

Disclosure of Invention

In view of the above, the present invention aims to provide an application of cydarifamide and its derivatives in preparing a radiotherapy sensitizer, so that the cydarifamide and its derivatives can reduce the risk of generating treatment resistance to tumors, enhance the tumor inhibition activity of radiotherapy, and show stronger tumor inhibition effect than that of single treatment when combined with radiotherapy.

The invention proves that the combination of the Xidabenamine and the radiation therapy can further inhibit the in vitro growth of the tumor cell culture and promote the apoptosis of the tumor cell culture through an in vitro cell model and an experimental animal model; meanwhile, the stem cell characteristics of tumor cells can be reduced by the independent or combined radiotherapy of the Sida benamine, and the independent treatment of the radiotherapy has no influence; in tumor-inoculated animal models, combined therapy with cydapamide radiation can produce a greater antitumor effect than either treatment alone. Therefore, the invention provides the application of the above-mentioned xidapamide and its derivatives in the preparation of radiotherapy sensitizers. Meanwhile, the invention also provides the application of the combination of the cidentamine and the derivatives thereof with radiotherapy in treating tumors or preparing products for treating tumors, wherein the products comprise but are not limited to medicaments.

The present invention relates to a novel compound of the present invention, and more particularly to a novel compound of the present invention, wherein the novel compound of the present invention is a novel compound of the present invention.

Preferably, the tumor includes, but is not limited to, lung cancer, liver cancer, stomach cancer, breast cancer, ovarian cancer, lymphoma, leukemia, colon cancer, rectal cancer, kidney cancer, pancreatic cancer, esophageal cancer, uterine cancer, vaginal cancer, prostate cancer, melanoma, urothelial cancer, and sarcoma.

In addition, the invention also provides a method for treating tumors, and particularly relates to the combined use of the cidentamine and the derivatives thereof and radiotherapy. Wherein the radiation therapy is performed simultaneously with or sequentially in any order with the sildenamine and its derivatives.

According to the technical scheme, the invention discloses that the cydariamine has the sensitization effect of anti-tumor radiotherapy, the combination of the cydariamine and the radiotherapy can more strongly inhibit the proliferation of tumor cells and induce the apoptosis of the cells, and simultaneously inhibit the stem cell characteristics of the tumor cells, and the combination of the cydariamine and the radiotherapy shows stronger tumor inhibition effect in animals compared with the single treatment. The cydarifamid not only can enhance the tumor inhibition activity of radiotherapy, but also can reduce the treatment resistance and recurrence risk of tumors, and the combination of the cydarifamid and the radiotherapy can provide better continuous curative effect than single treatment.

Drawings

FIG. 1 shows the results of comparing the relative proliferation rates of two model cells under different treatment conditions; wherein denotes a difference P <0.05 in combination treatment compared to radiotherapy treatment; indicates a difference P <0.01 for the combined treatment compared to the radiotherapy treatment;

FIG. 2 shows the comparison of the apoptosis of two model cells under different treatment conditions; wherein, denotes a difference P <0.01 of the cydapamide or the radiation treatment compared to the solvent control; # indicates a difference P <0.01 for the combination treatment compared to either xidabenamine or radiation treatment alone; # indicates that the difference P of the combined treatment compared to the sildenamine or radiotherapy treatment alone was < 0.001;

FIG. 3 shows a comparison of the formation of clonal cell masses by model cells under different treatment conditions;

FIG. 4 shows a comparison of the number of clonal cell masses formed by model cells under different treatment conditions; wherein, denotes a difference P <0.01 of the xidabenamine treatment compared to the solvent control; # indicates a difference P <0.01 for the combination treatment compared to the radiation treatment alone; # indicates a difference P <0.001 for the combination treatment compared to the radiotherapy treatment alone;

FIG. 5 shows a comparison of the average size (diameter) of the colony forming clones of model cells under different treatment conditions; wherein, denotes a difference P <0.01 of the xidabenamine treatment compared to the solvent control; # indicates a difference P <0.01 for the combination treatment compared to the radiation treatment alone;

FIG. 6 shows a comparison of the ratio of lactate dehydrogenase-positive stem cell-like tumor cells under different treatment conditions; wherein, denotes a difference P <0.01 of the xidabenamine treatment compared to the solvent control; denotes difference P <0.001 of the xidabenamine treatment compared to the solvent control; # indicates a difference P <0.01 for the combination treatment compared to the radiation treatment alone; # indicates a difference P <0.001 for the combination treatment compared to the radiotherapy treatment alone;

FIG. 7 is a graph showing the change in tumor volume at different time points in different treatment groups of a transplanted tumor animal model; wherein, represents the tumor volume difference P <0.01 of the xidabenamine or radiotherapy treatment compared to the control treatment; represents the tumor volume difference P <0.05 in the combined treatment group compared to the xidabenimine or radiation treatment alone;

FIG. 8 shows a comparison of tumor weights of different treatment groups after the tumor growth experiment in the transplanted tumor animal model is completed; wherein denotes a difference P <0.05 between the two groups; denotes the difference P <0.01 between the two groups.

Detailed Description

The invention discloses application of the cidamide and derivatives thereof, and can be realized by appropriately improving process parameters by persons skilled in the art with reference to the content. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the invention has been described in terms of preferred embodiments, it will be apparent to those skilled in the art that variations and modifications, as appropriate, may be made to the disclosed use of the technology herein described to practice and use the technology herein without departing from the spirit and scope of the invention.

In a specific embodiment, the activity difference of the combined therapy of the sidapipramine and the radiotherapy on the inhibition of the proliferation of tumor cells, the promotion of the apoptosis of tumor cells, the inhibition of the characteristics of tumor stem cells and the antitumor effect in animals compared with the two separate therapies is compared on the tumor cell line cultured in vitro and the animal model inoculated with the tumor cells. The experiments prove that the Sidapamide can enhance the antitumor activity of radiotherapy and provides the application of the Sidapamide in the sensitization of antitumor radiotherapy. In these experiments, the present invention selected a dose of either 300 nanomoles or 20 mg/kg body weight of Sidamide and a dose of 1-4 gray of radiation therapy.

The use of the provided cidentamine and its derivatives is further explained below.

Example 1: comparison of the effects of Simidamine in combination with radiation therapy on proliferation and apoptosis of tumor cell lines cultured in vitro compared to two treatments alone

And (3) experimental design: the single drug of the Sidapamide, the radiation therapy with different doses and the combined treatment of the Sidapamide and the radiation therapy are respectively adopted on different tumor cell lines cultured in vitro, and the influence of different treatment methods on the cell proliferation and the cell apoptosis is examined.

The test process comprises the following steps: the human lung cancer cell lines NCI-H2170 and NCI-H226 were cultured in vitro under the recommended culture conditions, and the control solvents dimethylsulfoxide (1/1000), cidalimide 300 nanomolar, radiation treatments at different doses of 1 gray, 2 gray, 4 gray, or both cidalimide and different doses of radiation treatments were added, respectively. After 24 hours of treatment, the number of viable cells was measured using a conventional CCK-8 kit (Shanghai macroloba), and the relative cell proliferation rate of each treatment group was calculated as compared to control solvent treatment. The results are shown in FIG. 1.

NCI-H2170 and NCI-H226 were cultured in vitro according to the recommended culture conditions, with the addition of the control solvents dimethyl sulfoxide (1/1000), Sidapamide 300 nanomolar, a single dose of 2 gray with radiation, or simultaneous treatment with both Sidapamide and radiation, respectively. After 24 hours of treatment, cell samples were collected for Annexin V and Propidium Iodide (PI) staining, and the staining of cells in different treatment groups was detected by flow cytometry (Bidy corporation), wherein the Annexin V singly stained cells were cells in the early apoptosis stage, and in the late apoptosis stage if doubly stained cells were detected, and the types and proportions of apoptotic cells in different treatments were statistically compared. The results are shown in FIG. 2.

And (3) test results: as shown in fig. 1, the single treatment of cidentamine at 300 nanomolar concentration showed only slight inhibition of the in vitro growth of both tumor cell lines, and the combined treatment with different doses of radiation significantly inhibited the relative proliferation rate of model cells compared to the single treatment with radiation, showing combined in vitro pro-inhibition.

As shown in FIG. 2, the treatment with Sidapamide at 300 nM and 2 Gray dose of radiotherapy promoted both early and late apoptosis in model cells. When the two treatments are combined, the cell apoptosis in the early stage or the late stage is more remarkably promoted compared with the single treatment.

The results of fig. 1 and 2 show that the cydariamine has the sensitization effect of the anti-tumor radiotherapy, and compared with the single treatment, the combination of the cydariamine and the radiotherapy can remarkably inhibit the in vitro proliferation of the tumor cells and induce the apoptosis of the tumor cells.

Example 2: effect of Simidamine in combination with radiation therapy on tumor Stem cell characteristics compared to two separate treatments

And (3) experimental design: one of the important reasons for the development of a tumor to be resistant to treatment (including radiation therapy, chemotherapy, targeted therapy or immunotherapy, etc.) or to relapse is the stem cell nature of the tumor cells, and a few, even a single tumor cell with stem cell characteristics, can self-renew and proliferate and reform a new tumor. Tumor stem cell characteristics can be assessed by the in vitro clonal (cell-mass) growth of individual tumor cells. The experiment examined the effect of different treatments on the ability of tumor cell clones (cell clusters) to form under in vitro culture conditions.

The test process comprises the following steps: the tumor cell lines NCI-H2170 and NCI-H226 were suspension cultured in low adsorption plates using different treatment conditions including solvent control (DMSO 1/1000), Sidapamine 300 nM, single dose 2 Gray in radiotherapy, and combination of Sidapamine 300 nM and 2 Gray in radiotherapy, and after 14 consecutive days of culture, the number, diameter, and stem cell-like tumor cell ratio of the colony formed were observed and compared for differences between different treatments. The results are shown in FIGS. 3-6.

And (3) test results: as shown in fig. 3, 4, and 5, compared to the control treatment, the sildenamine significantly reduced the number and diameter of tumor clonal cell masses at a dose of 300 nanomolar, whereas the radiation treatment had no effect on the above criteria, which was also significantly reduced by the combination of the sildenamine and the radiation treatment. The lactate dehydrogenase-positive tumor cells are considered to have the characteristics of tumor stem cells, namely, the cells can be self-renewed and form clone-like growth, as shown in fig. 6, the ratio of the lactate dehydrogenase-positive tumor cells can be remarkably reduced by the cydarifamide, the radiotherapy treatment has no obvious influence compared with the contrast treatment, and the ratio of the stem cell-like tumor cells can be remarkably inhibited by the cydarifamide combined radiotherapy treatment. In conclusion, the cydariamine can obviously inhibit the stem cell-like characteristics and the clonal growth of tumor cells, has the sensitization effect of the anti-tumor radiotherapy, can reduce the risks of tumor resistance and relapse after treatment by combining the radiotherapy, and improves the continuous curative effect.

Example 3: comparison of the Effect of Simidamine in combination with radiation therapy on tumor growth in animal models of transplantable tumors versus two separate treatments

And (3) experimental design: the humanized tumor cells are inoculated to a nude mouse to form tumor-like growth, and different experimental treatments are adopted to evaluate the influence of the humanized tumor cells on the in-vivo growth of the tumor.

The test process comprises the following steps: human tumor cell line NCI-H2170 was inoculated subcutaneously on both sides of BALB/c nude mice, and tumors were visualized and randomized into groups 14 days later, with administration of either xidalamine (20 mg/kg body weight once a day, suspension gavage), radiation (2 gray dose once a week), xidalamine combination radiation, or gavage solvent control, respectively. Model mice were measured continuously for body weight and tumor volume, and after six weeks the experiment was terminated, and the tumors were isolated and weighed. Experiments were conducted to examine the effect of different experimental treatments on tumor size and weight. The results are shown in FIGS. 7-8.

And (3) test results: as shown in fig. 7 and 8, sildendrine or radiotherapy alone had a slight inhibitory effect on tumor growth (tumor volume, weight), while the two combined showed a more significant inhibitory effect than the two treatments alone.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.