WO2023143611A1 - Method for treating cancer - Google Patents

Abstract

The present application relates to a method for treating cancer, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of formula I or an isotopic variant, a tautomer, a pharmaceutically acceptable salt, a solvate or a hydrate thereof. The present application further relates to use of a compound of formula I or an isotopic variant, a tautomer, a pharmaceutically acceptable salt, a solvate or a hydrate thereof for treating cancer. The cancer is colon cancer, brain glioma, liver cancer, intrahepatic/extrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, gastric/gastroesophageal junction adenocarcinoma, gastrointestinal stromal tumor, solid tumor, colorectal cancer, small cell/non-small cell lung cancer, local advanced or metastatic differentiated thyroid cancer/thyroid myeloid cancer, diffuse large B cell lymphoma, head, neck and chest tumor, primary malignant bone tumor, malignant melanoma, pancreatic neuroendocrine tumor, urothelial carcinoma, gastrinoma, cell tumor, insuloma, or cervical cancer.

Description

a way to treat cancer

This application claims the priority of the Chinese patent application with the application number 202210114156.7 and the title of the invention "a method for treating cancer" filed with the China Patent Office on January 30, 2022, the contents of which should be understood to be incorporated by reference In this application.

technical field

This article relates to the field of medicine, in particular to the use of a compound of formula I or its isotopic variants, tautomers, pharmaceutically acceptable salts, solvates or hydrates in the preparation of drugs for the treatment of cancer, and the treatment of cancer method.

Background technique

KDR2-2 is a small molecule compound, chemical name 2-(2-aminopyrimidin-4-yloxy)-N-(3-(trifluoromethyl)phenyl)quinoline-5-carboxamide, structure As shown in formula I

The synthesis method of compound KDR2-2 can refer to International Patent Publication WO2014/166386A1. International patent publication WO2021213512A1 discloses that the compound inhibits the abnormal proliferation of new blood vessels by inhibiting the activity of VEGFR2/KDR and PDGFRβ, and can be used to treat the occurrence of new blood vessels in the eye.

It is well known that the permeable blood-brain barrier formed between the blood and the brain effectively prevents more than 98% of small molecule drugs and almost 100% of large molecule drugs from exerting their therapeutic effects in the brain. Due to the effect of the blood-brain barrier, absolutely most of the drugs cannot reach the focal part of the nerve center, thereby failing to reach an effective therapeutic concentration and unable to exert the therapeutic effect of the drug.

The transport of small molecules across the BBB mainly consists of passive diffusion, efflux, and carrier-mediated endocytosis. The passive diffusion and efflux of drugs are mainly affected by the physicochemical properties of the molecules. Common strategies to improve the blood-brain barrier permeability of drug molecules and reduce the efflux rate include: increasing the lipophilicity of compounds, reducing hydrogen bond donors, deleting or replacing negatively charged atoms to reduce tPSA, removing basic groups to reduce pKa, And the introduction of constrained conformation to improve molecular rigidity is one of the important factors to improve the above effect.

Glioma refers to the tumor originating from the glial cells of the brain and is the most common primary intracranial tumor. The World Health Organization (WHO) classification of central nervous system tumors divides glioma into grades I-IV. Grades I and II are low-grade gliomas, and grades III and IV are high-grade gliomas.

The treatment of glioma is mainly based on surgical resection, combined with radiotherapy, chemotherapy and other comprehensive treatment methods. Surgery can relieve clinical symptoms, prolong survival, and obtain enough tumor samples for clear pathological diagnosis and molecular genetic testing. The principle of surgical treatment is to safely remove tumors in the largest range, and new technologies such as conventional neuronavigation, functional neuronavigation, intraoperative neurophysiological monitoring, and intraoperative MRI real-time images can help achieve safe tumor removal in the largest range.

Radiotherapy can kill or inhibit tumor cells and prolong the survival period of patients. Conventional fractionated external beam radiation is the standard treatment for glioma radiotherapy. Postoperative radiotherapy combined with temozolomide (TMZ) and adjuvant chemotherapy for glioblastoma (GBM) has become an Standard treatment regimen for adults with newly diagnosed GBM. The treatment of glioma requires the cooperation of multiple disciplines such as neurosurgery, neuroimaging, radiotherapy, neuro-oncology, pathology and neurorehabilitation. Following the principles of evidence-based medicine, individualized comprehensive treatment is adopted to optimize and standardize the treatment plan. In order to achieve the maximum therapeutic benefit, prolong the progression-free survival (PFS) and overall survival (OS) of patients as much as possible, and improve the quality of life. In order to enable patients to obtain the most optimal comprehensive treatment, doctors need to closely follow up and observe patients, conduct regular imaging review, and take into account patients' daily life, social and family activities, nutritional support, pain control, rehabilitation treatment and psychological regulation, etc. .

However, surgical resection and radiotherapy and chemotherapy have some drawbacks.

The degree of surgical resection is closely related to the prognosis of the disease. In low-grade glioma, total tumor resection or subtotal resection is better than partial resection or biopsy. Total resection or subtotal resection can not only prolong the survival time of patients, but also reduce the probability of glioma progression. A retrospective study of 1097 patients with low-grade glioma found that the median survival time was 10.5 years and 14 years for patients with less than 50% resection and 50%-99% resection, and patients with total resection, Median survival was over 15 years. However, glioma infiltrates the surrounding normal brain tissue, and it is difficult to find a clear resection boundary during surgery. In practice, it is difficult to find a balance between completely resecting the tumor and retaining more healthy tissue.

The side effects of radiation and chemotherapy vary from person to person. Factors such as tumor type, tumor location, dose, duration of treatment, and other factors all come into play. Side effects are cumulative, which means they get worse during treatment.

Contents of the invention

The inventors of the present application unexpectedly found that KDR2-2 has a high inhibitory efficiency for VEGFR2 receptors and can penetrate the blood-brain barrier.

In view of the unexpected discovery of the inventor, the application provides a compound of formula I

Use of the isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof in the preparation of a medicament for treating cancer.

The present application also provides a method for treating cancer, comprising administering a therapeutically effective amount of a compound represented by formula I or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or Hydrate.

The present application also provides the compound represented by formula I or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate for use in treating cancer.

In some preferred embodiments, the cancer is colon cancer, glioma, liver cancer, intrahepatic/extrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, gastric/gastroesophageal junction adenocarcinoma , gastrointestinal stromal tumor (GIST), solid tumors, colorectal cancer, small cell/non-small cell lung cancer, locally advanced or metastatic differentiated thyroid cancer/medullary thyroid carcinoma (MTC), diffuse large B-cell lymphoma, Head, neck, and thoracic tumors, primary malignant tumors of bone, malignant melanoma, pancreatic neuroendocrine tumors, urothelial carcinoma, gastrinoma, cytoma, insulinoma, or cervical cancer.

In some preferred embodiments, the cancer is colon cancer or glioma.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof inhibits tumor cell migration and growth.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof is administered parenterally, orally or ophthalmically.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof is administered by injection or orally.

Additional features and advantages of the application will be set forth in the description which follows, and, in part, will be obvious from the description, or may be learned by practice of the application. Other advantages of the present application can be realized and obtained through the schemes described in the specification and drawings.

Figure overview

The accompanying drawings are used to provide an understanding of the technical solution of the present application, and constitute a part of the specification, and are used together with the embodiments of the present application to explain the technical solution of the present application, and do not constitute a limitation to the technical solution of the present application.

Fig. 1 shows in embodiment 1, each group cell scratch pattern (100 *) at different times, wherein K is blank control group; 5 is 39ng/mL KDR2-2 group; 4 is 165ng/mL KDR2-2 group; 3 is 625ng/mL KDR2-2 group; 2 is 2500ng/mL KDR2-2 group; 1 is 10000ng/mL KDR2-2 group;

Figure 2 shows the average tumor volume for Example 2. Compared with the model control group in the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001; compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001; compared with the positive control group in the same period , ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001; compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

Figure 3 shows the mean relative tumor volumes for Example 2. Compared with the model control group in the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001; compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001; compared with the positive control group in the same period , ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001; compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

FIG. 4 shows the average tumor weight of Example 2. Compared with the model control group in the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001; compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001; compared with the positive control group in the same period , ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001; compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

FIG. 5 shows the average body weight of Example 2. Compared with the model control group in the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001; compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001; compared with the positive control group in the same period , ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001; compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

Fig. 6 shows MRI images of mouse brains treated with KDR2-2.

Figure 7 shows the lesion volume in mice treated with KDR2-2.

Figure 8 shows the body weight of mice treated with KDR2-2.

Fig. 9 shows that after treatment with KDR2-2, the hair of mice turns white.

detail

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any materials and methods similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred materials and methods are now described.

The application provides a compound of formula I

Use of the isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof in the preparation of a medicament for treating cancer.

The present application also provides a method for treating cancer, comprising administering a therapeutically effective amount of a compound represented by formula I or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or Hydrate.

The present application also provides the compound represented by formula I or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate for use in treating cancer.

In some preferred embodiments, the cancer is colon cancer, glioma, liver cancer, intrahepatic/extrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, gastric/gastroesophageal junction adenocarcinoma , gastrointestinal stromal tumor (GIST), solid tumors, colorectal cancer, small cell/non-small cell lung cancer, locally advanced or metastatic differentiated thyroid cancer/medullary thyroid carcinoma (MTC), diffuse large B-cell lymphoma, Head, neck, and thoracic tumors, primary malignant tumors of bone, malignant melanoma, pancreatic neuroendocrine tumors, urothelial carcinoma, gastrinoma, cytoma, insulinoma, or cervical cancer.

In some preferred embodiments, the cancer is colon cancer or glioma.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof inhibits tumor cell migration and growth.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof is administered parenterally, orally or ophthalmically.

In some preferred embodiments, the compound or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate thereof is administered by injection or orally.

In some embodiments, the present application provides a pharmaceutical composition comprising the compound of formula I or its isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate. The pharmaceutical composition includes a pharmacologically effective amount of the compound of formula I or its isotopic variants, tautomers, pharmaceutically acceptable salts, solvates or hydrates and pharmaceutically acceptable auxiliary materials. These excipients are known to those skilled in the art, for example, physiological saline, gelatin, gum arabic, lactose, microcrystalline cellulose, starch, modified starch, cellulose, modified cellulose, sodium glycolate , calcium hydrogen phosphate, magnesium stearate, talc, colloidal silicon dioxide, etc. In addition, these compositions may further contain: stabilizers, wetting agents, emulsifiers, sweeteners, flavoring agents, buffering agents and the like.

In some embodiments, the pharmaceutical composition provided by the application comprising the compound of formula I or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate can be formulated as Solid or liquid form for oral administration, such as tablet, pill, oral liquid, etc.; or, sterile solution, suspension or emulsion form, etc. for parenteral administration.

In some embodiments, the pharmaceutical composition provided by the application comprising the compound of formula I or its isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate is formulated as an ophthalmic preparation form.

In some embodiments, the compound of formula I of the present application or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate or its pharmaceutical composition can be used as a single pharmaceutical active ingredient to be administered, or may be administered in combination with other anticancer drugs.

In the present application, the "effective amount" or "effective dose" refers to an amount sufficient to affect beneficial or desired symptoms of the disease, its complications, or intermediate pathological indicators in the course of disease development.

In some embodiments, a compound of Formula I, or an isotopic variant, tautomer, pharmaceutically acceptable salt, solvate, or Hydrate or its pharmaceutical composition.

In some embodiments, based on the compound of formula I, the compound of formula I or its isotopic variants, tautomers, pharmaceutically acceptable salts, solvates or hydrates can be dosed at 0.24-640 mg/day administered to human subjects.

The compound of formula I mentioned herein, that is, KDR2-2 is a small molecular compound, chemical name 2-(2-aminopyrimidin-4-yloxy)-N-(3-(trifluoromethyl)phenyl ) quinoline-5-carboxamide, structure as shown in formula I

The present application will now be described with reference to the following examples, which illustrate some preferred aspects of the application. It should be understood, however, that the specificity of the following description herein does not supersede the generality of the preceding description herein, and that various other modifications and/or changes may be made, as disclosed herein, without departing from the spirit of the text.

Example 1: Effect of KDR2-2 on the Migration of Tumor Cells

In this example, the cell scratch test was used to study whether KDR2-2 has an inhibitory effect on the activity and movement of tumor cells.

Uncontrolled proliferation of tumor tissue, abnormal differentiation of tumor cells, and the ability of tumor cells to invade and metastasize are the most basic biological characteristics of malignant tumors, and invasion and metastasis are the main reasons why malignant tumors threaten the health and even life of patients. Therefore, it is of great significance to study the law of tumor invasion and metastasis and its mechanism for the prevention and treatment of malignant tumors.

In this example, a wound healing model of tumor cells was established in vitro, and the effect of KDR2-2 on the motility of HT-29 cells was studied by the cell scratch method.

experimental reagent

Dimethylsulfoxide (DMSO), Sigma;

McCoy's 5A medium (1×), Procell;

Pen Strep Glutamine (100×), Lot No.: 2051357, gibco;

Fetal Bovine Serum(FBS), Lot No.:11H144, Uruguay, ExCell Bio;

PBS, Lot No.: AE29456443, Hyclone;

MTS, Lot No.: 0000349697, PROMEGA;

0.25% trypsin-EDTA (trypsin), Lot No.: 1967360, gibco.

experiment equipment

0.22μm microporous membrane, R6SA39060, Millipore;

Centrifuge tubes (5mL, 10mL, 50mL), OTHBR, HX;

Tips (100μL, 1mL), Thermo QSP;

96-well culture plate, Thermo NUNC;

25T, 75T culture flask, Thermo NUNC;

Counting board, Tianjing;

Cover glass, Startech;

Pipette gun (100-1000 μL, 10-100 μL), Eppendorf;

Electronic balance, BSA124S, Sartorius;

Multiskan FC microplate reader, Thermo;

Inverted microscope, DMi1, Leica;

Three-hole electric heating constant temperature water tank, DK-8D, Shanghai Yiheng Scientific Instrument Co., Ltd.;

SW-CJ-1F clean bench, Suzhou Antai Air Technology Co., Ltd.;

Incubator, CCL-170B-8, ESCO;

High-pressure steam sterilizer, Shanghai Boxun YXQ-LS-50G.

Preparation of test article and culture medium

Complete medium: 89% McCoy’s 5A basal medium + 10% FBS + 1% double antibody Pen Strep Glutamine (100×);

cell line

HT-29 colon cancer cells.

experimental design

1. Grouping

Blank control group (1% DMSO) and different concentrations of KDR2-2 administration groups (10000ng/mL, 2500ng/mL, 625ng/mL, 156ng/mL, 39ng/mL).

2. Cell Recovery and Culture

Take out the frozen cells and place them in a 37°C water bath for rapid recovery (within 1 min);

Take out the cell solution and place it in a 10mL centrifuge tube, add 4 times the volume of complete medium, and centrifuge at 1000rpm for 5min;

Remove the supernatant, add 2mL complete medium to make cell suspension;

Transfer the cell liquid to a 25T culture flask, and make up the complete medium to a total volume of 5 mL;

Place the culture flask in an incubator (37°C, 5% CO ₂ ) for culture.

3. Cell passage

Subculture the cells when they reach 80% confluence;

Pour out the medium, add 1mL PBS to wash the cells twice;

Add 1mL trypsin to start digestion, observe under the microscope;

When the cell tentacles retract and become round (3min), add 2mL of complete medium to stop digestion;

Hold the culture flask with your left hand, and tap the side of the culture flask with your right hand to detach the cells;

Transfer the cell solution to a 10mL centrifuge tube and centrifuge at 1000rpm for 5min;

Remove the supernatant, add 1mL complete medium to make cell suspension;

Transfer the cell liquid to a 25T culture flask, and make up the complete medium to a total volume of 5 mL;

Place the culture flask in an incubator (37°C, 5% CO ₂ ) for culture.

4. Cell Counting Method

Dilute the prepared cell suspension 10 times with complete medium;

Add a cover slip, take 20 μL of the cell suspension and inject it from the side of the cover slip to spread the cell liquid, and use filter paper to absorb the excess cell liquid;

Place the counting plate under an inverted microscope to observe the number of cells in 4 large squares (each consisting of 1 small square). The principle of pressure line cell counting: count the left and not the right, count the top and not the bottom;

Cell number = (number of cells in 4 squares/4×10 ⁴ ×dilution factor) cells/mL;

According to the cell number measured for the first time, use complete medium to dilute to the cell number required for the experiment.

5. Cell Migration Assay

Adjust the cell density to 5×10 ⁵ cells/mL, add 1 mL per well into a 12-well plate for culture;

Use a ruler to draw two parallel straight lines at the bottom of the culture plate;

When the cell density is about 70%, use a 200 μL pipette tip to make a vertical and parallel line scratch in the middle of each well with the help of a ruler, and wash the cells with PBS 3 times with gentle movements to prevent the cells from falling off in pieces;

After washing, the corresponding concentration of drug solution (culture solution containing 1% FBS) was added to each well, and the pictures were recorded as 0h cell photos. Cultivate for 28 hours, and take a picture at the time point of 22 hours in the middle, and take a picture at the place where the parallel line connects with the "one" scratch, 2 pictures per well.

Use image J to calculate the scratch width, calculate the initial and end width ratio of each hole, and compare between groups.

Experimental results

The scratch width ratios of HT-29 cells in each group at different time points are shown in Table 1, and the scratch diagrams of cells in each group at different time points are shown in Figure 1 (100×).

Table 1 Ratio of scratch width of HT-29 cells in each group at different time points

Note: Compared with the blank control group, *p<0.05, there is a statistical difference; **p<0.01, there is a significant difference.

It can be seen from Table 1 that compared with the blank control group, 10000ng/mL KDR2-2 could significantly inhibit the migration of HT-29 cells at 22h (p<0.01); 2 can significantly inhibit the migration of HT-29 cells (p<0.01).

It can be seen from Figure 1 that the cells in the 10000ng/mL and 2500ng/mL KDR2-2 groups did not proliferate and the cells were in poor condition as the incubation time increased. produce.

Example 2: Efficacy evaluation of KDR2-2 on HT-29 human colon cancer tumor model

In this example, a colon cancer model was established by inoculating HT-29 cells subcutaneously, and then intervened with KDR2-2 to study whether KDR2-2 can inhibit the growth of colon cancer.

Preparation of the test product and the test product solution

KDR2-2 suspension (specification: 0.4mL: 0.4mg; 1mL: 20mg), as the test product.

The specification is 0.4mL: 0.4mg of KDR2-2 suspension contains 0.10% (w/v) KDR2-2, 0.40% (w/v) hypromellose (HPMC E4M), 4.50% (w/v) Polyethylene glycol 15-hydroxystearate ( HS 15), 0.21% (w/v) sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ 2H ₂ O), 1.60% (w/v) disodium hydrogen phosphate dodecahydrate (Na ₂ HPO ₄ 12H ₂ O), 0.30% (w/v) sodium chloride, 0.10% (w/v) anhydrous sodium thiosulfate, make up the volume with water for injection.

The specification is 1mL: 20mg of KDR2-2 suspension containing 2% (w/v) KDR2-2, 0.40% (w/v) hypromellose (HPMC E4M), 4.50% (w/v) 15- Polyethylene glycol hydroxystearate ( HS 15), 0.21% (w/v) sodium dihydrogen phosphate dihydrate (NaH ₂ PO ₄ 2H ₂ O), 1.60% (w/v) disodium hydrogen phosphate dodecahydrate (Na ₂ HPO ₄ 12H ₂ O), 0.30% (w/v) sodium chloride, 0.10% (w/v) anhydrous sodium thiosulfate, make up the volume with water for injection.

10mg/mL KDR2-2 suspension: Take an appropriate amount of 20mg/mL KDR2-2 suspension and add an equal volume of blank excipients to dilute to obtain the target concentration solution.

Positive control substance and preparation

For mitomycin, use 1 mL sodium chloride injection to dissolve mitomycin lyophilized powder into a 10 mg/mL solution (stock solution). Take an appropriate amount of the stock solution and dilute it 200 times with sodium chloride injection to obtain a 0.05 mg/mL mitomycin solution.

Main Reagents and Drugs

Main Experiment Consumables

Main Instruments and Apparatus

experiment system

Experimental animals: SPF grade BALB/c nude mice, 5-6 weeks old, weighing 15-18g. Purchased from Guangdong Medical Experimental Animal Center; production license number: SCXK (Guangdong) 2018-0002; animal quality certificate number: No.44007200077964.

Each animal is identified by a tail tag and a cage card. The marking method of the cage card during the environmental adaptation period: indicate the experiment number, animal strain, cage number, number of animals, time of entry, animal number, etc. Marking method of the cage card after grouping: Indicate the experiment number, animal species, treatment factors, number of animals, cage number, start and end time of the experiment, animal number, etc. on the cage card.

The mice were acclimated to the environment for 6 days before modeling. The main inspection contents of the adaptation period: whether it is consistent with the quality indicators required at the time of order; general status; whether the body weight reaches the weight range required by the experiment; the adaptation period inspection is qualified, and subcutaneous inoculation is carried out to make models.

Stocking density: 5 animals/cage during the environmental adaptation period, 5 animals/cage for the formal experiment.

experimental design

Cell Recovery and Culture

Take out the cryopreserved HT-29 human colon cancer cells and place them in a 37°C water bath for quick recovery (within 1 min);

Take out the cell solution and place it in a 10mL centrifuge tube, add 4 times the volume of complete medium (the complete medium here is the same as in Example 1), and centrifuge at 1000rpm for 5min;

Remove the supernatant, add 2mL complete medium to make cell suspension;

Transfer the cell liquid to a 25T culture flask, and make up the complete medium to a total volume of 5 mL;

Place the culture flask in an incubator (37°C, 5% CO ₂ ) for culture.

cell inheritance

Subculture the cells when they reach 80% confluence;

Pour out the medium, add 1mL PBS to wash the cells twice;

Add 1mL trypsin to start digestion, observe under the microscope;

When the cell tentacles retract and become round (3min), add 2mL of complete medium to stop digestion;

Hold the culture flask with your left hand, and tap the side of the culture flask with your right hand to detach the cells;

Transfer the cell solution to a 10mL centrifuge tube and centrifuge at 1000rpm for 5min;

Remove the supernatant, add 2mL complete medium to make cell suspension;

Transfer the cell liquid to a 25T culture flask, and make up the complete medium to a total volume of 5 mL;

Place the culture flask in an incubator (37°C, 5% CO ₂ ) for culture.

Cell Counting Method

Dilute the prepared cell suspension 10 times with complete medium;

Add a cover slip, take 20 μL of the cell suspension and inject it from the side of the cover slip to spread the cell liquid, and use filter paper to absorb the excess cell liquid;

Place the counting plate under an inverted microscope to observe the number of cells in 4 large squares (each consisting of 1 small square). The principle of pressure line cell counting: count the left and not the right, count the top and not the bottom;

Cell number = (number of cells in 4 squares/4×10 ⁴ ×dilution factor) cells/mL;

Cell preparation for seeding

The HT-29 human colon cancer cell line was cultured and passaged routinely, after three passages in vitro. Centrifuge to obtain cells according to the above operation, resuspend the cells in the basal medium, absorb 10 μL of cells and dilute them 10 times, and calculate ^the cell concentration. Adjust the cell concentration to 1×10 ⁷ cells/mL.

Subcutaneous xenograft tumor model construction

Place the cell suspension on ice, blow it gently to avoid the generation of air bubbles, mix slowly, grab the nude mouse, make it sideways, disinfect the axillary skin of the forelimb with iodophor, draw 100 μL of the cell suspension with a 1mL syringe and inject Under the skin of the armpit, withdraw the needle slowly to prevent cells from overflowing.

Animal grouping and dosage

When the tumor volume reached 100mm ³ , random grouping was performed according to the tumor volume. See the table below for grouping and dosing information:

Table 2 group administration information table

animal grouping

Group design: model group, solvent control group, positive control group, KDR2-2 high and low dose group subcutaneous injection (10mg/kg, 100mg/kg), KDR2-2 high and low dose group intragastric administration (10mg/kg, 100mg/kg) .

Number of animals: 5 animals per group except the model group 6;

Grouping method: select tumor-forming animals for experiments, and randomly group them according to tumor volume;

Treatment of remaining animals: After grouping, the mice that did not meet the experimental requirements were euthanized.

Dosing Information

Administration route: gavage, subcutaneous and intraperitoneal

Dosing frequency: 1 time/day

Administration volume: 0.1mL/10g

The day of the first administration is defined as the first day of the experiment (day1, D1)

Observation and inspection

General Status Observation

Observation time: observe once a day after tumor inoculation, and the observation frequency can be appropriately increased according to the actual situation;

Routine observation content: tumor growth, body weight, appearance and signs of mice, general behavioral activities, mental state, respiratory state, feces properties, genitalia, death, etc.;

Observed animals: all experimental animals.

weight

Measurement time: measure once before the first administration, and measure twice a week during the administration period;

Measuring animals: all surviving experimental mice.

tumor size

Observation time: Tumor size was measured twice a week;

Measuring method: measure the long diameter (a) and short diameter (b) of the transplanted tumor with a vernier caliper;

Observed animals: all tumor-bearing animals before grouping, and all surviving experimental animals after grouping.

animal anatomy

Dissection time: D20

Dissected Animals: All planned dissected animals.

Tumor weight detection: The animals were sacrificed on the second day after the last administration, and the complete tumor body was carefully removed, weighed and recorded with an electronic balance.

Tumor fixation: After the tumor was weighed, a portion of the tumor was selected and fixed in 10 times the volume of neutral formaldehyde.

Treatment Effect Evaluation

Relative tumor proliferation rate T/C (%)

The formula for calculating tumor volume (V) is: V=a×b ² ×l/2;

Among them, a and b represent the length and width respectively. The relative tumor volume (RTV) was calculated according to the measured results, and the calculation formula was: RTV=V _t /V ₀ . Where V ₀ is the tumor volume measured on the day of group administration (ie, the first day), and V _t is the tumor volume at each measurement. The evaluation index of antitumor activity is the relative tumor proliferation rate T/C (%), and the calculation formula is as follows: T/C (%)=T _RTV /C _RTV × 100%; T _RTV : treatment group RTV; C _RTV : model control Group RTV.

Evaluation criteria for curative effect: T/C(%)>40% is invalid; T/C(%)≤40%, and P<0.05 compared with the model group by variance analysis is effective.

Calculation of tumor inhibition rate (%)

The tumor inhibition rate can be calculated by tumor weight or tumor volume.

At the end of the experiment, the animals were euthanized, and the tumor pieces were dissected to weigh the tumor weight of each group;

Calculate the tumor inhibition rate (%) according to the following formula:

Tumor growth inhibition rate%=(average tumor weight of the control group-average tumor weight of the treatment group)/average tumor weight of the control group×100%;

Tumor growth inhibition rate % (calculated by tumor volume):

If T>T0, tumor growth inhibition rate (TGI)%=[1-T/C]×100%; if T<T0, tumor growth inhibition rate (TGI)%=[1-(T-T0)/T0] ×100%. Where T and C are the tumor volumes of the treatment group and the control group at the end of the experiment, respectively; T0 and C0 are the tumor volumes of the treatment group and the control group at the beginning of the experiment, respectively.

Criteria for judging effectiveness: tumor inhibition rate (ie tumor growth inhibition rate) < 40% is invalid; tumor inhibition rate ≥ 40% and P < 0.05 through statistical processing is effective.

Comprehensive Judgment Standard

It is effective if one of the above two effective criteria (relative tumor proliferation rate and tumor inhibition rate) meets.

Statistical Analysis

All data are presented as mean ± SD. First, use Levene's Test to test the homogeneity of the data. If the data is uniform (p>0.05), then perform one-way analysis of variance; if the analysis of variance is significant (p≤0.05), then perform Dunnett's multiple comparisons, and analyze between drug groups Do not comment. If the result of Levene's Test is significant (p≤0.05), the Kruskal-Wallis nonparametric test was performed. If the result of Kruskal-Wallis non-parametric test is significant (p≤0.05), the Mann-Whitney U test will be used for pairwise comparison.

Experimental results

1. Tumor volume:

Model control group: on D0 (15 days after inoculation and modeling, group administration began), D7 (7 days after group administration), D14, and D20, the tumor volumes were 181.21±67.42mm ³ , 498.14±136.65mm ³ , 796.39±121.96mm mm ³ and 1161.48±168.3mm ³ .

Vehicle control group: on D0, D7, D14 and D20, the tumor volumes were 172.7±57.27mm ³ , 625.95±222.89mm ³ , 1202.55±509.6mm ³ and 1387.18±506.07mm ³ , compared with the same period model at D7, D14 and D20 There was no statistical difference between the groups (p>0.05).

Positive control group: on D0, D7, D14 and D20, the tumor volumes were 198.54±88.3mm ³ , 722.09±179.94mm ³ , 990.64±260.59mm ³ and 745.18±108.36mm ³ . On D7 and D14, there was no statistical difference compared with the model control group and the solvent control group during the same period (p>0.05); on D20, the tumor volume decreased compared with the model control group and the solvent control group during the same period, and there was a significant statistical difference ( p≤0.05).

In the low dose group of the test product: on D0, D7, D14 and D20, the tumor volumes were 182.96±71.85mm ³ , 435.54±154.92mm ³ , 672.77±234.09mm ³ and 923.84±325.26mm ³ . There was no statistical difference (p>0.05) on D7 compared with the model control group and the solvent control group at the same period (p>0.05). Compared with the model control group and the positive control group, there was no statistical difference (p>0.05), and compared with the solvent control group in the same period, the tumor volume was reduced and there was a significant statistical difference (p≤0.05); There was no statistical difference between the control group and the positive control group (p>0.05).

In the high-dose group of the test product: on D0, D7, D14 and D20, the tumor volumes were 180.67±52.45mm ³ , 299.38±110.7mm ³ , 302.72±130.68mm ³ and 250.29±83.45mm ³ . On D7, there was no statistical difference (p>0.05) compared with the model control group and the low-dose group of the test product in the same period, and compared with the solvent control group and the positive control group in the same period, the tumor volume was all reduced and there was a significant statistical difference (p≤ 0.01, p≤0.001); on D14 and D20, compared with the model control group, solvent control group, positive control group and low-dose test product group, the tumor volumes were all reduced and there were significant statistical differences (p≤0.01, p ≤0.01, p≤0.01, p≤0.001).

In conclusion, intraperitoneal injection of mitomycin and KDR2-2 suspension can inhibit the tumor volume of HT-29. Among them, 5mg/kg mitomycin can significantly inhibit or even reduce the tumor volume. The inhibitory effect of KDR2-2 suspension on tumor volume can be seen in a certain dose-response relationship. The low dose of 10mg/kg KDR2-2 suspension The inhibitory effect is slightly weak, and the high dose of 100mg/kg KDR2-2 suspension has a very strong inhibitory effect.

The average tumor volume results of each group are shown in Figure 2 and Table 3.

Table 3 Average tumor volume (mm ³ , Mean±SD)

Note: n indicates the number of animals included in the statistical analysis.

Compared with the model control group at the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001.

Compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001.

Compared with the positive control group in the same period, ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001.

Compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

2. Relative tumor volume (RTV):

Model control group: On D7, D14 and D20, the RTVs were 2.92±0.74, 4.84±1.67 and 7.78±1.93, respectively.

Solvent control group: On D7, D14 and D20, the RTVs were 3.73±0.97, 6.91±1.24 and 8.19±1.79, respectively, and there was no statistical difference compared with the model control group during the same period (p>0.05).

Positive control group: on D7, D14 and D20, the RTVs were 4.00±1.20, 5.86±3.34 and 4.21±1.93, respectively. Compared with the model control group and the solvent control group in the same period on D7 and D14, there was no statistical difference (p>0.05); compared with the model control group and the solvent control group in the same period, the RTV on D20 decreased and there was a significant statistical difference (p ≤0.05).

In the low dose group of the test product: on D7, D14 and D20, the RTVs were 2.44±0.40, 3.86±1.15 and 5.4±2.46, respectively. On D7, there was no statistical difference compared with the model control group and the same period (p>0.05), and compared with the solvent control group and the positive control group during the same period, the RTV was reduced and there was a significant statistical difference (p≤0.05, p≤0.05) ; There was no statistical difference (p>0.05, p>0.05) compared with the model control group and the solvent control group at the same period on D14, and the RTV was reduced compared with the positive control group and there was a significant statistical difference (p≤0.05) ; There was no statistical difference in D20 compared with the model control group, solvent control group and positive control group (p>0.05).

In the high-dose group of the test product: on D7, D14 and D20, the RTVs were 1.64±0.19, 1.63±0.3 and 1.32±0.33, respectively. On D7 and D4, compared with the model control group, solvent control group, positive control group and test product low-dose group, the RTV all decreased and had significant statistical differences (p≤0.01, p≤0.01, p≤0.01, p≤0.001), on D20, compared with the model control group, solvent control group and test product low-dose group, the RTV all decreased and had significant statistical differences (p≤0.001, p≤0.001, p≤0.01), Compared with the positive control group in the same period, there was no statistical difference (p>0.05).

In conclusion, intraperitoneal injection of mitomycin and KDR2-2 suspension both have inhibitory effects on HT-29 relative tumor volume (RTV). 5mg/kg mitomycin can significantly reduce RTV, and the inhibitory effect of KDR2-2 suspension on the relative tumor volume can be seen in a certain dose-response relationship, and the inhibitory effect of low-dose 10mg/kg KDR2-2 suspension is weak , high-dose 100mg/kg KDR2-2 suspension has a very strong inhibitory effect, which is consistent with its inhibitory effect on tumor volume.

The average relative tumor volume (RTV) results of each group are shown in Figure 3 and Table 4.

Table 4 Mean Relative Tumor Volume (Mean±SD)

Note: n indicates the number of animals included in the statistical analysis.

Compared with the model control group at the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001.

Compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001.

Compared with the positive control group in the same period, ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001.

Compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

3. Relative tumor proliferation rate T/C (%):

Solvent control group: at D7, D14 and D20, T/C were 127.88%, 142.87% and 105.33%, respectively.

Positive control group: on D7, D14 and D20, T/C were 137.03%, 121.19% and 54.12%, respectively.

The low dose group of the test product: on D7, D14 and D20, T/C were 83.20%, 80.17% and 69.42% respectively.

In the high-dose group of the test product: on D7, D14 and D20, T/C were 56.73%, 34.03% and 16.98% respectively.

According to T/C (%) > 40% is invalid; T/C (%) ≤ 40%, and compare p<0.05 to be effective curative effect evaluation standard with model group through analysis of variance, positive control drug mitomycin ( 5mg/kg) and high-dose test product KDR2-2 suspension (100mg/kg) all reach effective evaluation standard, show that mitomycin and KDR2-2 suspension all can effectively inhibit HT-29 tumor under this condition proliferation.

4. Tumor weight:

At the end of the experiment, all surviving animals were euthanized, the tumor mass was dissected and weighed, and the average tumor weight of each group was calculated.

The average tumor weight of the model control group was 0.82±0.23g.

The average tumor weight of the solvent control group was 0.92±0.25g, which was not significantly different from that of the model control group (p>0.05).

The average tumor weight of the positive control group was 0.47±0.05g, which was smaller than that of the model control group and the solvent control group with significant statistical difference (p≤0.05, p≤0.01).

The average tumor weight of the low-dose group of the test product was 0.63±0.16g, which had no statistical difference compared with the model control group and the positive control group (p>0.05), and decreased compared with the solvent control group and had statistically significant There were no statistical differences (p≤0.05).

The average tumor weight of the high-dose group of the test product was 0.18±0.05g, which was significantly reduced compared with the model control group, the solvent control group, the positive control group and the low-dose group of the test product (p≤0.001 , p≤0.001, p≤0.05, p≤0.001).

In summary, intraperitoneal injection of 5 mg/kg mitomycin and KDR2-2 suspension (10 mg/kg and 100 mg/kg) both have inhibitory effects on the tumor weight of HT-29 tumors. Among them, 100mg/kg KDR2-2 suspension has the strongest inhibitory effect on tumor weight, followed by 5mg/kg mitomycin, and 10mg/kg KDR2-2 suspension is slightly weaker. The inhibitory effect of KDR2-2 suspension on the relative tumor volume shows a certain dose-response relationship. This is consistent with its inhibitory effect on tumor volume and relative tumor volume results.

The results of average tumor weight in each group are shown in Figure 4 and Table 5.

Table 5 Average tumor weight (g, Mean±SD)

Note: n indicates the number of animals included in the statistical analysis.

Compared with the model control group at the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001.

Compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001.

Compared with the positive control group in the same period, ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001.

Compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

5. Tumor inhibition rate:

Solvent control group: the tumor inhibition rate was -5.33% (calculated by tumor volume) and -11.65% (calculated by tumor weight).

Positive control group: tumor inhibition rate was 45.88% (calculated by tumor volume) and 43.50% (calculated by tumor weight).

The low-dose group of the test product: the tumor inhibition rate was 30.58% (calculated by tumor volume) and 23.99% (calculated by tumor weight).

The high-dose group of the test product: the tumor inhibition rate was 83.02% (calculated by tumor volume) and 78.39% (calculated by tumor weight).

According to tumor inhibition rate (i.e. tumor growth inhibition rate) < 40% is invalid; tumor inhibition rate ≥ 40% and after statistical processing p < 0.05 is an effective efficacy evaluation standard, positive control drug mitomycin (5mg/kg) and high-dose test product KDR2-2 suspension (100mg/kg) all reached the effective evaluation standard, showing that under this condition, both mitomycin and KDR2-2 suspension can effectively inhibit the proliferation of HT-29 tumors.

6. Weight:

Model control group: on D0, D7, D14 and D20, the body weight was 21.96±1.45g, 22.48±1.58g, 21.44±2.25g and 21.08±3.15g.

Solvent control group: on D0, D7, D14 and D20, the body weight was 23.02±0.99g, 22.94±0.88g, 22.9±0.94g and 22.42±0.87g, compared with the model control group at the same period on D0, D7, D14 and D20 There was no statistical difference (p>0.05).

Positive control group: on D0, D7, D14 and D20, the body weight was 21.54±0.27g, 20.42±0.94g/18.52±0.97g and 17.7±1.41g. Compared with the model control group and the solvent control group in the same period at D7, there was a significant statistical difference (p≤0.05, p≤0.01); at D14, compared with the model control group and the solvent control group in the same period, the And there is a significant statistical difference (p≤0.01, p≤0.001); compared with the model control group at the same time, there is a decrease and a significant statistical difference (p≤0.05) at D20, but the death of the animal makes the group There are only two individual data, so the statistical data is not very meaningful.

The low-dose group of the test product: on D0, D7, D14 and D20, the body weight was 22.01±1.23g, 21.79±1.23g, 21.97±1.54g and 21.45±1.55g. Compared with the same period model control group, solvent control group and positive control group, there was no statistical difference (p>0.05) at D7; there was no statistical difference compared with the same period model control group and solvent control group at D14 and D20 (p >0.05), compared with the positive control group in the same period, the body weight increased and there was a significant statistical difference (p≤0.001, p≤0.05). Similarly, the sample size of the positive control group was too small at D20, and the statistical significance was not significant.

The high-dose group of the test product: on D0, D7, D14 and D20, the body weight was 22.29±1.11g, 20.21±1.18g, 17.29±1.17g and 15.03±0.46g. Compared with the same period of model control group, solvent control group and test product low dose group, the body weight all decreased and there was a significant statistical difference (p≤0.01, p≤0.001, p≤0.05) at D7, compared with the same period of positive control group There was no statistical difference (p>0.05); when D14 and D20, compared with the model control group, the solvent control group, and the low-dose group of the test product, the body weight all decreased and there was a significant statistical difference (p≤0.001, p ≤0.001, p≤0.001), there was no statistical difference compared with the positive control group in the same period (p>0.05).

To sum up, the intraperitoneal injection of the positive control drug mitomycin and the high-dose test product 100mg/kg KDR2-2 suspension can significantly reduce the body weight of the animals. In the positive control group (mitomycin 5 mg/kg), 3 animals died on the 6th day of administration, accounting for 60% of the animals in this group.

The average body weight results of each group are shown in Figure 5 and Table 6.

Table 6 Average body weight (g, Mean±SD)

Note: n indicates the number of animals included in the statistical analysis.

Compared with the model control group at the same period, ^a p≤0.05, ^aa p≤0.01, ^aaa p≤0.001.

Compared with the solvent control group in the same period, ^b p≤0.05, ^bb p≤0.01, ^bbb p≤0.001.

Compared with the positive control group in the same period, ^c p≤0.05, ^cc p≤0.01, ^ccc p≤0.001.

Compared with the low dose group of the test product in the same period, ^d p≤0.05, ^dd p≤0.01, ^ddd p≤0.001.

in conclusion

Under the conditions of this experiment, intraperitoneal injection of KDR2-2 suspension for 20 days can significantly and effectively inhibit Inhibit the growth and proliferation of HT-29 tumors, and show a certain dose-response relationship.

Example 3: Preliminary evaluation of the curative effect of KDR2-2 on glioma

model building

Experimental animals and dosing settings

Adult male C57BL/6 mice (7-8 weeks) were purchased from the Experimental Animal Center of Sun Yat-sen University, and KDR2-2 was administered 2 weeks after gamma knife irradiation (to induce glioma formation). The doses of 10 and 80 mg/kg body weight were continuously administered for 4 weeks.

Preparation of the test product and the test product solution

KDR2-2 suspension (specification: 0.4mL: 4.0mg, the preparation is the same as in Example 2).

The vehicle control without drug was placebo.

animal grouping

Group design: model group, solvent control group, KDR2-2 high and low dose group subcutaneous injection (80 mg/kg, 10 mg/kg). The preparation of KDR2-2 test sample is the same as in Example 2. KDR2-2-L represents the KDR2-2 low-dose group, and KDR2-2-H represents the KDR2-2 high-dose group.

Number of animals: 10 animals/group in the model group, 10 animals/group in the solvent control group, and 10 animals/group in each of the high and low dose groups.

Dosing Information

Administration route: intraperitoneal injection; administration frequency: 1 time/day

Indicator detection

MRI detection: detect the size of edema focus at 1w, 2w, 4w, and 6w after irradiation

Experimental results

Fig. 6 shows MRI images of mouse brains treated with KDR2-2.

Figure 7 shows the lesion volume in mice treated with KDR2-2.

Figure 8 shows the body weight of mice treated with KDR2-2.

Figure 9 demonstrates graying of mouse hair after treatment with KDR2-2.

The application describes a number of embodiments, but the description is illustrative rather than restrictive, and it will be obvious to those of ordinary skill in the art that within the scope of the embodiments described in the application, There are many more embodiments and implementations. Although many possible combinations of features are shown in the drawings and discussed in the detailed description, many other combinations of the disclosed features are possible. Except where expressly limited, any feature or element of any embodiment may be used in combination with, or substituted for, any other feature or element of any other embodiment.

Claims (9)

1. Use of a compound of formula I or its isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate in the preparation of a medicament for treating cancer
   
2. The use according to claim 1, wherein the cancer is colon cancer, glioma, liver cancer, intrahepatic/external cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, stomach/gastroesophageal junction Adenocarcinoma, gastrointestinal stromal tumor (GIST), solid tumor, colorectal cancer, small cell/non-small cell lung cancer, locally advanced or metastatic differentiated thyroid cancer/medullary thyroid carcinoma (MTC), diffuse large B cell Lymphoma, head, neck and thoracic tumor, primary malignancy of bone, malignant melanoma, pancreatic neuroendocrine tumor, urothelial carcinoma, gastrinoma, cell tumor, insulinoma or cervical cancer; preferably, the cancer is colon cancer or glioma.
3. A method for treating cancer, comprising administering a therapeutically effective amount of a compound represented by formula I to a patient in need
   

   Or its isotope variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate.
4. The method according to claim 3, wherein the cancer is colon cancer, glioma, liver cancer, intrahepatic/external cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, stomach/gastroesophageal junction Adenocarcinoma, gastrointestinal stromal tumor (GIST), solid tumor, colorectal cancer, small cell/non-small cell lung cancer, locally advanced or metastatic differentiated thyroid cancer/medullary thyroid carcinoma (MTC), diffuse large B cell Lymphoma, head, neck and thoracic tumor, primary malignancy of bone, malignant melanoma, pancreatic neuroendocrine tumor, urothelial carcinoma, gastrinoma, cell tumor, insulinoma or cervical cancer; preferably, the cancer is colon cancer or glioma.
5. The method of claim 4, wherein the cancer is colon cancer, and the compound inhibits tumor cell migration and growth.
6. The method according to any one of claims 3-5, wherein the administration is parenteral, oral or ophthalmic.
7. The method according to any one of claims 3-5, wherein the administration is injection administration or oral administration.
8. Compound shown in formula I
   

   or its isotopic variants, tautomers, pharmaceutically acceptable salts, solvates or hydrates for use in the treatment of used in cancer.
9. The compound for use according to claim 8 or its isotopic variant, tautomer, pharmaceutically acceptable salt, solvate or hydrate, wherein the cancer is colon cancer, glioma, Liver cancer, intrahepatic/extrahepatic cholangiocarcinoma, gallbladder cancer, biliary tract cancer, kidney cancer/renal cell carcinoma, gastric/GEJ junction adenocarcinoma, gastrointestinal stromal tumor (GIST), solid tumors, colorectal cancer, small cell / non-small cell lung cancer, locally advanced or metastatic differentiated thyroid cancer / medullary thyroid carcinoma (MTC), diffuse large B-cell lymphoma, head, neck and thoracic tumors, primary malignant tumors of bone, malignant melanoma, pancreatic neuroendocrine tumors, Urothelial carcinoma, gastrinoma, cell tumor, insulinoma or cervical cancer; preferably, the cancer is colon cancer or glioma.