CN112807305A - Application of CH223191 in inhibiting migration of tumor cells - Google Patents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
- A61K31/415—1,2-Diazoles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
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Abstract
The invention relates to application of CH223191 in inhibiting cell migration, in particular to application of CH223191 or a pharmaceutical composition containing CH223191 in preparing a medicament for inhibiting glioma cell migration. The invention discovers for the first time that CH223191 with a specific concentration can inhibit the migration of a glioblastoma cell line U87, and provides a new choice for the treatment of glioblastoma.
Description
Technical Field
The present invention relates to the field of molecular biology, more specifically, the present invention relates to techniques related to cancer therapy. The invention mainly comprises the application of CH223191 in inhibiting the migration of tumor cells, and particularly discloses the application of CH223191 in inhibiting the migration of glioblastoma cells.
Background
In all cancer-related case studies, brain and central nervous system tumors are the third leading cause of cancer-related death. In brain tumors, the incidence of astrocytoma is high, accounting for 15.1% of primary central nervous system tumors and 46.1% of primary central nervous system malignant tumors. Among them, glioblastoma is the most common, belonging to a very aggressive primary glioma, and is also the IV grade tumor in the malignancy classification of World Health Organization (WHO) tumors.
The comprehensive treatment of glioblastoma includes surgery, radiotherapy, systemic treatment (chemotherapy, targeted therapy) and supportive treatment, but the overall prognosis is still poor and the long-term survival rate is low. There remains a need in the art for more therapeutic approaches for glioblastoma.
Disclosure of Invention
Studies have shown that there is diffuse growth of glioblastoma and, in addition to intracranial short-range migration, extracranial metastasis of tumor cells is also found. Therefore, inhibiting the migratory motor of glioblastoma may be beneficial for treating tumors, reducing the risk of postoperative recurrence and improving disease prognosis.
CH223191 is a synthetic compound that antagonizes important signaling pathways in the organism and has the structure shown below. The CH223191 can effectively inhibit the migration capability of glioblastoma, but the CH223191 with the same concentration has very low cytotoxicity, so that the CH223191 can be a drug for inhibiting the migration capability of glioblastoma, and has potential application values in the aspects of reducing the postoperative recurrence risk and improving the disease prognosis.
CH223191 structural formula
Accordingly, in one aspect, the present invention provides the use of CH223191 or a pharmaceutical composition comprising CH223191 in the preparation of a medicament for inhibiting glioma cell migration.
In some embodiments, the glioma cell is an astrocytoma cell.
In some embodiments, the glioma cell is a glioblastoma cell.
In some embodiments, the glioblastoma cell is a cell from a subject or a glioblastoma cell line.
In some embodiments, the subject has a glioblastoma.
In some embodiments, the glioma cell line is glioblastoma cell line U87.
In some embodiments, the concentration of CH223191 is 10-7～10-5M, e.g. 10-7M～10-6M、10-7M～10- 5M or 10-6M～10-5M。
In another aspect, the invention provides the use of CH223191 or a pharmaceutical composition comprising CH223191 in the manufacture of a medicament for the treatment of a glioma.
In some embodiments, the medicament is for inhibiting glioma growth, metastasis and/or invasion.
In some embodiments, the glioma is an astrocytoma.
In some embodiments, the glioma is a glioblastoma.
In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and optionally other therapeutic agents.
In some embodiments, the additional therapeutic agent is selected from temozolomide, bevacizumab, lomustine, carmustine, carboplatin, procarbazine, and vincristine.
In another aspect, the present invention provides a method of inhibiting glioma cell migration comprising the step of contacting the cell with an effective amount of CH223191 or a pharmaceutical composition comprising CH 223191.
In some embodiments, the glioma cell is an astrocytoma cell.
In some embodiments, the glioma cell is a glioblastoma cell.
In some embodiments, the glioblastoma cell is a cell from a subject or a glioblastoma cell line.
In some embodiments, the subject has a glioblastoma.
In some embodiments, the glioma cell line is glioblastoma cell line U87.
In some embodiments, the concentration of CH223191 is 10-7～10-5M, e.g. 10-7M～10-6M、10-7M～10- 5M or 10-6M～10-5M。
In some embodiments, the method is performed in vivo.
In some embodiments, the method is performed in vitro.
In another aspect, the present invention provides a method of treating a glioma, comprising the step of administering to a subject in need thereof a therapeutically effective amount of CH223191 or a pharmaceutical composition comprising CH 223191.
In some embodiments, the medicament is for inhibiting glioma growth, metastasis and/or invasion.
In some embodiments, the glioma is an astrocytoma.
In some embodiments, the glioma is a glioblastoma.
In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and optionally other therapeutic agents.
In some embodiments, the additional therapeutic agent is selected from temozolomide, bevacizumab, lomustine, carmustine, carboplatin, procarbazine, and vincristine.
Definition of terms
Molecular biology, microbiology, and recombinant DNA techniques that may be used in the present invention are within the skill of the art. These techniques have been explained fully in the literature. See, for example: sambrook, Fritsch & Maniatis, Molecular cloning: a Laboratory Manual, (1982); DNA Cloning, A Practical Approach Volumes I & II, D.N. Glover. 1985; oligonucleotide Synthesis, m.j.gait ed.1984; nucleic Acid Hybridization, B.D.Hames & S.J.Higgins eds.1985; transformation and transformation, B.D.Hames & S.J.Higgins eds.1984; animal Cell Culture, r.i. freshney, ed.1986; immobilized Cells And Enzymes, IRL Press, 1986; BPerbal, A Practical Guide To Molecular Cloning,1984 based thereon, the terms appearing herein are defined as follows.
As used in the specification and the appended claims, the singular forms "a", "an" and "the" include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to "a molecule" optionally includes combinations of two or more such molecules, and the like.
As used herein, the term "pharmaceutically acceptable carrier" refers to an ingredient in a pharmaceutical formulation other than an active ingredient that is not toxic to a subject. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers, preservatives, and the like.
As used herein, the term "individual" or "subject" is a mammal, e.g., a bovine, equine, porcine, canine, feline, rodent, primate; among these, particularly preferred subjects are humans.
As used herein, the term "effective amount" refers to an amount sufficient to obtain, or at least partially obtain, a desired effect. For example, a therapeutically effective amount refers to an amount sufficient to cure or at least partially arrest the disease and its complications in a patient already suffering from the disease. It is well within the ability of those skilled in the art to determine such effective amounts. For example, an amount effective for therapeutic use will depend on the severity of the disease to be treated, the general state of the patient's own immune system, the general condition of the patient, e.g., age, weight and sex, the mode of administration of the drug, and other treatments administered concurrently, and the like.
The amount of the drug administered to the subject depends on the type and severity of the disease or condition and the characteristics of the subject, such as general health, age, sex, body weight and tolerance to the drug, as well as on the type of formulation and mode of administration of the drug, and the period or interval of administration. One skilled in the art will be able to determine the appropriate dosage based on these and other factors.
As used herein, the terms "glioma", "glioma" refer to tumors derived from neuroepithelium and include primarily astrocytomas, oligodendrogliomas, ependymomas and the internodal (malignant) ependymomas, mixed gliomas, choroid plexus tumors, and the like, by pathological classification. "glioblastoma" is the most malignant glioma among astrocytomas.
Drawings
FIG. 1 shows 10-7～10-5M CH223191 treated U87 cells for 24h and 48h enzyme labeling instrument to detect the change of absorbance value of cell sample.
FIG. 2 shows the results of scoring experiments to evaluate the effect of CH223191 on the migration ability of U87 cells. Wherein FIG. 2A shows a warp beam 10-5After the M CH223191 treats the cells for 36 hours, the area of scratches among the cells is larger than that of a control group, which shows that the migration distance of the cells is reduced and the migration capacity of the cells is reduced; FIG. 2B shows a warp 10-5After treatment with M CH223191, the migration distance of U87 cells decreased by 24.0%, indicating that CH223191 was effective in inhibiting cell migration.
Detailed Description
Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
U87 cells were purchased from cell resource center of Chinese academy of medical sciences
CH223191 from Sigma
CCK8 kit was purchased from Sangon Biotech, Inc. (Shanghai)
Microscope: CKX41(Olympus Japan)
A camera: 600D (Canon Japan)
An enzyme-labeling instrument: tecan 2000pro (Tecan Switzerland)
Example 1 investigation of the toxic Effect of CH223191 on glioblastoma
U87 cells were cultured at 1X 105one/mL was seeded into 96-well plates at 100mL cell suspension per well. Overnight cultures awaited cell attachment. The 96-well plate was removed, blotted clean of the original medium, and washed once with Phosphate Buffered Saline (PBS). The experiment was set up with a background control group, a solvent control group and a drug treatment group. Corresponding low serum medium (serum concentration 1%, no cells in background control wells), solvent-containing dimethyl sulfoxide (DMSO) or varying concentrations of CH223191 was added to each well, maintaining the volume of solvent at 0.1%. And putting the medicated 96-well plate into an incubator to continuously culture for 24 hours or 48 hours. After the end of incubation time was reached, 10. mu.L of CCK8 reagent was added to each well as written in the kit instructions, and the well plate was again placed in the incubator and incubated for 1h before reading the absorbance with the microplate reader.
As can be seen in FIG. 1, use 10-7-10-5M CH223191 treated U87 cells did not cause significant changes in absorbance values within 48h, and had no effect on cell proliferation and viability. Therefore, CH223191 is not cytotoxic at this concentration and treatment time and can be used in subsequent experiments.
Example 2 inhibition of cell migration of U87 by CH223191
The migration ability of the cells was evaluated using a scratch test. The scratch was located by drawing 3 lines transversely across the bottom of the 6 well plate with a marker pen. Cells were plated at 4X 1052mL of cells were seeded at density per mL in 6-well plates and cultured overnight to wait for the cells to adhere. After the cells completely covered the bottom of the well plate, the cell monolayer was scratched with the tip of a 1mL pipette tip to form a scratch. And selecting the junction of the scratch and the mark drawn by the mark pen as a photographing position. The floating cells were removed by washing with phosphate buffer and then a low serum medium containing the solvent DMSO or CH223191 was added. And collecting the image by a digital camera under an inverted microscope 0-48h after scratching. Totally, three independent experiments are carried out, each concentration group in each independent experiment has 3 repeated holes, each hole comprises 3 scratches, and 12 images are shot for carrying outCounting and quantifying. Cell migration distance was calculated by dividing the scratch reduction area by the scratch length and analyzed using Image Pro Plus 6.0 software. The scale bar on the representative scratch image is 0.05 mm.
As shown in fig. 2A, after CH223191 treated the cells for 36h, the area of the scratches between the cells was larger than that of the control group, indicating that the migration distance of the cells was decreased and the migration ability of the cells was decreased. The statistical data according to FIG. 2B show that-5Migration distance of U87 cells decreased by 24.0% after M CH223191 treatment.
Although specific embodiments of the invention have been described in detail, it will be appreciated by those skilled in the art that, based upon the overall teachings of the disclosure, various modifications and alternatives to those details could be developed and still be encompassed by the present invention. The full scope of the invention is given by the appended claims and any equivalents thereof.
Claims (10)
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103179968A (en) * | 2010-07-27 | 2013-06-26 | 波士顿大学管理委员会 | Aryl hydrocarbon receptor (AhR) modulators as novel cancer therapies |
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| CN107496924A (en) * | 2017-08-31 | 2017-12-22 | 中国人民解放军第四军医大学 | Application and anti-tumor compositions of the AhR inhibitor in antineoplastic is prepared |
| US20200179341A1 (en) * | 2016-11-04 | 2020-06-11 | Ohio State Innovation Foundation | Methods and compositions for treating multiple myeloma and increasing antibody dependent cell cytotoxicity by targeting the aryl hydrocarbon receptor |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103179968A (en) * | 2010-07-27 | 2013-06-26 | 波士顿大学管理委员会 | Aryl hydrocarbon receptor (AhR) modulators as novel cancer therapies |
| CN104023713A (en) * | 2011-09-07 | 2014-09-03 | 德国癌症研究中心 | Means and methods for treating and/or preventing natural ahr ligand-dependent cancer |
| US20200179341A1 (en) * | 2016-11-04 | 2020-06-11 | Ohio State Innovation Foundation | Methods and compositions for treating multiple myeloma and increasing antibody dependent cell cytotoxicity by targeting the aryl hydrocarbon receptor |
| CN107496924A (en) * | 2017-08-31 | 2017-12-22 | 中国人民解放军第四军医大学 | Application and anti-tumor compositions of the AhR inhibitor in antineoplastic is prepared |
Non-Patent Citations (7)
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