KR20200061822A - Use of pyrimidinedione derivatives having apoptosis of brain cancer stem cells - Google Patents

Abstract

The present invention relates to novel use of a pyrimidinedione derivative compound having an activity of inducing apoptosis of brain tumor stem cells, and particularly to a pharmaceutical composition for destructing brain tumor stem cells and a pharmaceutical composition for preventing or treating brain tumor, which include 5[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound as an active ingredient, and a method for inducing apoptosis of brain tumor stem cells using the same. Particularly, since the compound can inhibit lipid metabolism of brain tumor stem cells and is highly effective for accelerating apoptosis of brain tumor stem cells, it can be used as a novel brain tumor-treating agent targeting brain tumor stem cells, and can be used widely as a medicine for assisting anticancer treatment in order to increase anticancer treatment efficiency of a brain tumor therapy.

Description

Use of pyrimidinedione derivatives having apoptosis of brain cancer stem cells}

The present invention relates to the use of a pyrimidinedione derivative compound having apoptosis ability of brain tumor stem cells, specifically 5[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene-2-thioxodihydro-4 ,6(1H,5H)-pyrimidinedione The present invention relates to a composition for killing brain tumor stem cells comprising an active ingredient.

A brain tumor refers to a tumor formed in a brain tissue or a membrane tissue surrounding it, and refers to a primary brain tumor and a secondary brain tumor that has metastasized to a brain tissue or a brain membrane from another site. Brain tumors found in adults include glioma, glioblastoma, meningioma (meningioma), pituitary adenoma (adenomas), and glioma, among which glioblastoma starts from glioma cells of the brain tissue. This is a tumor that accounts for 12-15% of all brain tumors and is the most common primary brain tumor in adults.

These brain tumors are typical refractory solid cancers with little improvement in survival rate over the past 30 years, and despite the recent advanced combination treatment technique, post-operative anti-cancer-radiation combination therapy, the average survival rate of brain tumor patients has evolved to 12 to 15 months. There is no situation.

On the other hand, non-specific anti-cancer treatment, which is a method of removing rapidly growing cells, has been presented with many side effects and does not effectively remove cancer cells. Since the development of Imatinib (Gleevec), a BCR-ABL kinase inhibitor specifically expressed for cancer, has been shown to have a good effect on leukemia patients, the concept of'targeted therapy' was introduced, and targeted therapeutics are tumor cells Since it targets only the main factors involved in the properties of'selectively', it has more side effects and is more effective in treatment than conventional non-cancer drugs.

Targeted therapies are targeted therapies that target activated phosphorylase or tumor-induced signaling systems (RTK/RAS/PI3K) in cancer, such as Gefitinib (Iressa), Sorafenib (Nexavar), and Bevacizumab (Avastin) since Imatinib was developed. It has been developed steadily. However, since the therapeutic effect of these targeted therapies is limited, patients are still forced to receive treatment using conventional non-specific anticancer agents such as TMZ. In addition, despite the use of these targeted therapies, many studies have reported that cancer eventually recurs, and in particular, in brain tumors, the targeted treatments developed to date have not been reported to improve the prognosis of patients at all.

Meanwhile, since cancer stem cells are first isolated from leukemia, cancer stem cells have been separated and identified as various cancer stem cells such as breast cancer stem cells, brain tumor stem cells, and colon cancer stem cells, and cancer stem cells are characterized by normal stem cells. It has been reported that phosphorus has autoantigenative ability, multidifferentiation ability, cell proliferation ability, chromosomes and genetic alterations characteristic of cancer cells, tumorigenesis, generation of differentiated cancer cells and high anticancer agent resistance. Therefore, even if multiple signaling mechanisms related to the formation of cancer are complexly inhibited, cancer stem cells cannot be prevented from recurring unless they are targeted, so existing cancer treatment methods can derive the fundamental therapeutic effect of cancer. Can't.

Therefore, in order to more effectively and fundamentally treat brain tumors, there is an urgent need to develop new targeted therapies for brain tumor stem cells.

Republic of Korea Registered Patent No. 10-1872950

Accordingly, the present inventors are able to inhibit lipid metabolism of brain tumor stem cells by effectively inhibiting SREBP1, which is known to play a major role in lipid metabolism and tumor signaling in brain tumor stem cells in which pyrimidinedione derivative compounds are more differentiated than brain tumor cells. The present invention was completed by clarifying that the compound can be used as a therapeutic agent for brain tumors through the death of brain tumor stem cells.

Accordingly, the object of the present invention is 5- [1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione (5 -[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound or a pharmaceutically acceptable salt thereof, as an active ingredient , To provide a pharmaceutical composition for killing brain tumor stem cells.

Another object of the present invention is a 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound or It is to provide a pharmaceutical composition for the prevention or treatment of brain tumors, which includes a pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound Or it is to provide a brain tumor treatment adjuvant containing the pharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H, It is to provide a method for inducing apoptosis of brain tumor stem cells, comprising the step of treating a 5H)-pyrimidinedione compound.

In order to achieve the object of the present invention as described above, the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H ,5H)-pyrimidinedione (5-[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound or a pharmaceutical thereof Provided is a pharmaceutical composition for killing brain tumor stem cells, comprising an acceptable salt as an active ingredient.

In one embodiment of the present invention, the compound may be to reduce the lipid metabolism by inhibiting the expression of SREBP1 in brain tumor stem cells.

In one embodiment of the present invention, the brain tumor is anaplastic astrocytomas, glioblastomas, meningiomas, pituitary tumors, schwannomas, central nervous system lymphoma. ), oligodendrogliomas, ependymomas, low-grade astrocytomas, medulloblastomas, astrocytic tumors, pylocytic astrocytoma , Diffuse astrocytomas, polymorphic xanthoastrocytomas, upper and lower giant cell astrocytomas, anaplastic oligodendrogliomas, oligoastrotomas, oligoastro Anaplastic oligoastrocytomas, myxopapillary ependymomas, subpendymomas, ventricular cell tumors (ependymomas), anaplastic ependymomas, and astroblastomas astroblastomas, chordoid gliomas of the third ventricle, gliomatosis cerebris, glangliocytomas, connective tissue-generating infantile astrocytomas, binding Tissue-producing infantile gangliogliomas, dysplastic neuroepithelial dysplasia (dyse) mbryoplastic neuroepithelial tumors, central neurocytomas, cerebellar liponeurocytomas, paragangliomas, epidenymoblastomas, tentative primitive ectodermal tumors, supratentorial primitive neuroectodermal tumors Choroids plexus papilloma, pineocytomas, pineoblastomas, moderate parenchymal tumors of intermediate differentiation, hemangiopericytomas, saddle area tumors It may be one or more selected from the group consisting of tumors of the sellar region, craniopharyngioma, capillary hemangioblastoma, and primary CNS lymphoma.

In addition, the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound or its It provides a pharmaceutical composition for the prevention or treatment of brain tumors, comprising a pharmaceutically acceptable salt as an active ingredient.

In addition, the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound or its Provided is a brain tumor treatment supplement comprising a pharmaceutically acceptable salt as an active ingredient.

Furthermore, the present invention provides 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)- to brain tumor stem cells in vitro. It provides a method for inducing apoptosis of brain tumor stem cells, comprising the step of treating a pyrimidinedione compound.

The 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound of the present invention is a brain tumor stem cell It can effectively reduce lipid metabolism by suppressing the expression of SREBP1, and has excellent activity to promote apoptosis of brain tumor stem cells, so it can be used as a novel brain tumor treatment targeting brain tumor stem cells and at the same time It can also be used in various ways as an anti-cancer drug to increase anti-cancer efficiency.

Figure 1 shows the 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H) of the present invention in five types of brain tumor stem cell lines. )- After treating the pyrimidinedione compound by concentration, the degree of apoptosis in each cell line is represented by a linear graph.
FIG. 2 shows 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H) of the present invention in five types of brain tumor stem cell lines. )- After treating the pyrimidinedione compound by concentration, the degree of apoptosis in each cell line is represented by a bar graph.
FIG. 3 shows 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H) of the present invention in five types of brain tumor stem cell lines. )- After treating the pyrimidinedione compound, it shows the result of confirming the level of SREBP1 expression in each cell line by Western blot.

In the present invention, 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound is a brain tumor stem cell. It is characterized in that it was first identified that it has the activity of promoting cell death.

According to an embodiment of the present invention, after treating the above compounds of the present invention for each concentration of five types of brain tumor stem cells by concentration, the degree of cell death of brain tumor stem cells was analyzed. As a result, the compounds of the present invention were not treated. Compared to the non-group, it was found that the degree of apoptosis was significantly superior in the brain tumor stem cell line treated with the compound of the present invention, and this apoptosis effect was observed in all five types of brain tumor stem cell lines.

Furthermore, in another embodiment, after treating the compound of the present invention with respect to five types of brain tumor stem cell lines, the degree of expression change of SREBP1 known to play an important role in lipid metabolism in brain tumor stem cells and affect the signaling mechanism of cancer. Analysis.

As a result, it was found that the expression of SREBP1 was effectively suppressed in all five brain tumor stem cells treated with the compound of the present invention compared to the group not treated with the compound.

On the other hand, according to recently reported in silico analysis results of gene expression data of brain tumor stem cells, undifferentiated brain tumor stem cells are known to have more active lipid synthesis metabolism compared to differentiated brain tumor cells, particularly SREBP1 (sterol regulatory) element-binding protein 1) is known as a regulator of fat synthesis metabolism, and recently it is known to play an important role in cancer metabolism and tumor signaling.

In this regard, if the lipid synthesis metabolism of brain tumor stem cells can be effectively suppressed, the growth, proliferation and activity of brain tumor stem cells can be inhibited, and ultimately, brain tumors can be prevented or treated as targets of brain tumor stem cells.

Accordingly, the present inventors confirmed that the compound of the present invention can effectively inhibit SREBP1 expression as a result of analyzing whether the tumor tumor stem cells have inhibitory activity against SREBP1, and thus inhibit the lipid metabolism of cells by suppressing the expression of SREBP1. It was found that the activity of brain tumor stem cells can be reduced or inhibited.

Accordingly, the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound (5- Brain tumor containing [1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) or a pharmaceutically acceptable salt thereof as an active ingredient It may provide a pharmaceutical composition for stem cell death and a pharmaceutical composition for the prevention or treatment of brain tumors.

The 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound according to the present invention is preferably It may be a compound having a structural formula of formula (1).

<Formula 1>

The compound of the present invention can be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid and aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes It is obtained from non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids. These pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulphite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, iodide Id, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitro benzoate, hydroxybenzoate, meth Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycol Rate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate or mandelate.

In the present invention, the term'brain tumor' refers to all tumors occurring in the skull, and includes all tumors occurring in the brain and structures surrounding the brain.

Specifically, the brain tumor is not limited thereto, but anaplastic astrocytomas, glioblastomas, meningiomas, pituitary tumors, schwannomas, central nervous system lymphoma (CNS lymphoma) , Oligodendrogliomas, ependymomas, low-grade astrocytomas, medulloblastomas, astrocytic tumors, pilocytic astrocytoma, Diffuse astrocytomas, polymorphic xanthoastrocytomas, upper and lower giant cell astrocytomas, anaplastic oligodendrogliomas, oligoastrocytomas, oligoastrocytomas Anaplastic oligoastrocytomas, myxopapillary ependymomas, subependymomas, ventricular cell tumors (ependymomas), anaplastic ependymomas, astroblastomas ), 3rd ventricle chordoid gliomas of the third ventricle, gliomatosis cerebris, glangliocytomas, connective tissue-generating infantile astrocytomas, connective tissue Producing infantile gangliogliomas, genital dysplasia neuroepithelialoma (d ysembryoplastic neuroepithelial tumors, central neurocytomas, cerebellar liponeurocytomas, paragangliomas, epidenymoblastomas, mammary primitive ectodermal tumors, supratentorial primitive neuroectodermal tumors Choroids plexus papilloma, pineocytomas, pineoblastomas, moderate parenchymal tumors of intermediate differentiation, hemangiopericytomas, saddle area tumors tumors of the sellar region, craniopharyngioma, capillary hemangioblastoma, or primary CNS lymphoma.

In the present invention,'brain tumor stem cells', a kind of cancer stem cells, are cells important for the growth and metastasis of brain tumors, and have the ability to maintain undifferentiated state, self-renewal and differentiation ability, and may generate tumors. In addition, it is distinguished from other groups and generates new tumors, so it exists in a small amount in cancer, but it can be said to be a key target for brain tumor treatment because it causes cancer recurrence and metastasis.

The composition of the present invention may include a pharmaceutically acceptable additive, pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, Mannitol, syrup, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opadry, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, white sugar Etc. can be used.

The pharmaceutically acceptable additive according to the present invention is preferably contained in 0.1 to 90 parts by weight based on the composition, but is not limited thereto.

In the present invention, the composition may be administered in various formulations, oral or parenteral, during actual clinical administration. When formulated, diluents such as fillers, bulking agents, binders, wetting agents, disintegrating agents, surfactants, etc., which are usually used, or It may be prepared using excipients, and suitable formulations known in the art are preferred to use those disclosed in Remington's Pharmaceutical Science, recently Mack Publishing Company, Easton PA. Carriers, excipients and diluents that may be included in the composition include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, mineral oil and the like.

The solid preparation for oral administration includes tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient such as starch, calcium carbonate, sucrose or It is prepared by mixing Lactose and gelatin. In addition, lubricants such as magnesium styrene talc are used in addition to simple excipients. In addition, the liquid formulations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used, various excipients, such as wetting agents, sweeteners, fragrances, preservatives, etc. This can be included.

The formulation for parenteral administration includes sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories.

Non-aqueous solvents, suspension solvents include propylene glycol (Propylene glycol), polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin butter, and glycerogelatin may be used. The parenteral administration may be performed using an external skin or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, or intrathoracic injection.

The composition of the present invention may be administered in a pharmaceutically effective amount.

The'pharmaceutically effective amount' refers to an amount sufficient to treat the disease at a reasonable benefit/risk ratio applicable to medical treatment, and the effective dose level is the individual type and severity, age, sex, the type of virus, the drug It can be determined by activity, sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including co-drugs, and other factors well known in the medical arts.

The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And it can be administered single or multiple. Considering all of the above factors, it is important to administer an amount that can achieve the maximum effect in a minimal amount without side effects, and can be easily determined by those skilled in the art.

The term'administration' as used in the present invention means to provide the composition of the present invention to a subject in any suitable way.

In the present invention, the composition may be administered to a subject by various routes. Any mode of administration can be expected, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrathecal dura or cerebral vascular injection.

In the present invention, the composition can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers for the inhibition or death of brain tumor stem cells.

In the present invention,'prevention' refers to all actions that inhibit brain tumors or delay the onset of the disease by administration of the composition.

In the present invention,'improvement' refers to all actions that improve or advantageously alter symptoms of brain tumors by administration of the composition.

In the present invention,'treatment' refers to the act of alleviating brain tumors by administration of the composition.

In the present invention,'metastasis' refers to a condition in which a brain tumor, a malignant tumor, spreads from the brain to other tissues such as lungs or stomach.

In addition, the present invention is 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound or its It is possible to provide a brain tumor treatment supplement comprising a pharmaceutically acceptable salt as an active ingredient.

In the present invention, the term'brain tumor treatment supplement' refers to a material that can be used as an auxiliary to enhance the effect of a brain tumor treatment method commonly used in the art, and by using the mixed extract according to the invention of the brain tumor treatment method It can enhance the effect.

In the present invention, the types of brain tumor treatments that can be used with the brain tumor supplement include brain surgery, radiation therapy, chemotherapy using anti-cancer agents, and the like, and a combination of two or more of the above-described treatments Can be More specifically, the brain tumor treatment method may preferably be concurrent chemoradiotherapy (CCRT) or a combination treatment of brain surgery and CCRT, but is not limited thereto.

In addition, the brain tumor treatment method may be selected under general principles to be considered when selecting a brain tumor treatment method such as the type of cancer cell, the rate of absorption of the anticancer agent (the treatment period and the route of administration of the anticancer agent), the location of the tumor, and the size of the tumor.

Furthermore, the present invention can provide a method for inducing apoptosis of brain tumor stem cells using the compound of the present invention, that is, 5-[1-(4-hydroxyphenyl)-1H-pyrrole in brain tumor stem cells in vitro. -2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound (5- [1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2 -thioxodihydro-4,6(1H,5H)-pyrimidinedione) to induce apoptosis of the brain tumor stem cells.

Therefore, the 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound of the present invention has a brain tumor stem. Since it can prevent, improve, and treat brain tumors by targeting cells, it can be used as a new therapeutic agent for brain tumors, and at the same time, it can be used in various ways as an anticancer adjuvant drug to increase the anticancer efficiency of brain tumor treatment.

 Hereinafter, the present invention will be described in more detail through examples. These examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited to these examples.

<Preparation example>

Sample and cell culture

The brain tumor stem cells used in the following examples are brain tumor stem cell lines derived from brain tumor patients, 528NS (glioblastoma, Ichiro Nakano, University of Alabama at Birmingham), MD13 (glioblastoma, University of Alabama at Birmingham), GSC11 (glioblastoma, Joshua) M. Shulman, Texas Children's Hospital), GSC23 (glioblastoma, Joshua M. Shulman, Texas Children's Hospital), GSC20 (glioblastoma, Joshua M. Shulman, Texas Children's Hospital) were used. Each cell is a brain tumor stem cell culture condition 1% antimicrobial agent (apenicillin/streptomycin, Lonza, Basel, Switzerland), 0.04% B27 (Invitrogen, MA, USA), epidermal growth factor (20 ng/mL; R&D Systems, MN, USA) and DMEM/F12 media (Hyclone, GE healthcare life sciences, IL, USA) culture medium containing basic broblast growth factor (20 ng/mL; R&D Systems) at 37° C., 5 % CO ₂ conditions and used.

In addition, 5-[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione (ChemBridge Corporation, San Diego, CA, USA) compounds It was dissolved in DMSO to a final concentration of 10 mM and used.

< Example 1>

Of the present invention Pyrimidinedione Effect of derivative compound on cell death of brain tumor stem cells

The present inventors showed that 5-[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound, which is a pyrimidinedione derivative compound, of brain tumor stem cells The following experiment was carried out to confirm whether there is an apoptosis effect.

To this end, the brain tumor stem cell lines 528NS, MD13, GSC11, GSC23, and GSC20 cells were respectively dispensed into a 96-well plate (SPL, Korea) to be 5,000 cells/well, and after 24 hours, 5-[1-(4-hydroxyphenyl )-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound is treated by concentration (0, 0.05, 0.1, 0.5, 1, 5, 10 μM), and then , Incubated for 3 days. Thereafter, 20 μL per well was added and the reaction reagent of the Ez-Cytox kit (DOGEN, Seoul, Korea) was added and reacted for 4 hours, followed by a PowerWaveXS microplate-reader (Bio-Tek Instruments Inc., VT, USA) device at a wavelength of 450 nm. By measuring the absorbance by using, the degree of apoptosis according to the concentration of compound treatment for each cell line was measured. At this time, the measurement values were described as mean±SEM.

As a result, as shown in FIGS. 1 and 2, the compounds of the present invention were found to have apoptosis effects on all five types of brain tumor stem cells, and the survival rate of the cells was not treated with the compounds of the present invention. Compared to the present invention, it was found to be significantly reduced when treating the compound of the present invention.

Therefore, through these results, the present inventors have a pyrimidinedione derivative compound, 5-[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione compound Was confirmed to have apoptosis ability of brain tumor stem cells, and it was found that the compound can be used as a new brain tumor treatment targeting brain tumor stem cells.

< Example  2>

According to the invention Pyrimidinedione Derivative compounds in brain tumor stem cells SREBP1  Inhibitory effect

Next, the present inventors analyzed whether the compounds of the present invention have inhibitory activity against SREBP1, which plays an important role in lipid metabolism and cancer signaling for brain tumor stem cells.

To this end, the brain tumor stem cell lines 528NS, MD13, GSC11, GSC23, and GSC20 cell lines were dispensed at 2.0×10 ⁶ cells/9 mL, and after 24 hours, the compound of the present invention was diluted in 1 mL of the culture solution to give a final concentration of 10 mL. After treating to 10 μM, cells were collected after 72 hours, washed with PBS, precipitated at 3500 rpm, and lysed by adding RIPA solution to the precipitated cell pellet. Precipitated at 15000 rpm to collect only the supernatant to obtain protein concentration. It was measured. Protein was transferred to the membrane by loading 15 μg of protein per sample. Then, the membrane was blocked using 5% skim milk powder, and the primary antibody was reacted at 4°C for 12 hours. Anti-SREBP1 (sc-8984, Santa Cruz Biotechnology, TX, USA) and anti-β-actin (sc-47774, Santa Cruz Biotechnology) were used as antibodies. After washing 3 times for 5 minutes using TBST, horseradish peroxidase (HRP)-coupled secondary antibody (Vector Laboratories, CA, USA) was added and reacted for 1 hour at room temperature, and then 3 times for 5 minutes using TBST. After washing, the ECL (EBP-1073, ELPIS biotech. Inc., Korea) solution was reacted for 1 minute, and then developed by reacting with a film in the dark.

As a result of the analysis, as shown in FIG. 3, it was confirmed that protein expression of SREBP1 was inhibited in all five brain tumor stem cells treated with the compound of the present invention.

Through these results, the compound of the present invention can be used as a new brain tumor therapeutic agent targeting brain tumor stem cells by reducing the lipid metabolism of brain tumor stem cells and ultimately inducing apoptosis by inhibiting the expression and activity of SREBP1 in brain tumor stem cells. And it was found.

So far, the present invention has been focused on the preferred embodiments. Those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered in terms of explanation, not limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent range should be interpreted as being included in the present invention.

Claims (6)

5- [1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione (5- [1-(4 -hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound or a pharmaceutically acceptable salt thereof as an active ingredient, for the death of brain tumor stem cells Pharmaceutical composition. According to claim 1,
The compound is characterized in that to reduce the lipid metabolism by inhibiting the expression of SREBP1 in brain tumor stem cells, brain tumor stem cell death pharmaceutical composition. According to claim 1,
The brain tumors are anaplastic astrocytomas, glioblastomas, meningiomas, pituitary tumors, schwannomas, central nervous system lymphoma (CNS lymphoma), oligodendrogliomas , Ependymomas, low-grade astrocytomas, medulloblastomas, astrocytic tumors, pilocytic astrocytoma, diffuse astrocytomas, Pleomorphic xanthoastrocytomas, upper and lower giant cell astrocytomas, anaplastic oligodendrogliomas, oligoastrocytomas, anaplastic oligocytomas, anaplastic astrocytomas ), myxopapillary ependymomas, subependymomas, ependymomas, anaplastic ependymomas, astroblastomas, and third ventricle axons Chordoid gliomas of the third ventricle, gliomatosis cerebris, glangliocytomas, connective tissue-generating infantile astrocytomas, connective tissue-generating infantile gangliogliomas ), dysbryoplastic neur oepithelial tumors, central neurocytomas, cerebellar liponeurocytomas, paragangliomas, epidenymoblastomas, tentative primitive neuroectodermal tumors, choroid plexus (choroids plexus papilloma), pineocytomas, pineoblastomas, moderate parenchymal tumors of intermediate differentiation, hemangiopericytomas, tumors of the saddle area of the sellar region), craniopharyngioma, capillary hemangioblastoma, and primary CNS lymphoma, characterized in that at least one selected from the group consisting of, pharmaceutical composition for brain tumor stem cell killing . 5- [1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione (5- [1-(4 Prevention or treatment of brain tumors comprising -hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound or a pharmaceutically acceptable salt thereof as an active ingredient Dragon pharmaceutical composition. 5- [1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione (5- [1-(4 -Hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound or a pharmaceutically acceptable salt thereof as an active ingredient, brain tumor treatment adjuvant. 5-[1-(4-hydroxyphenyl)-1H-pyrrole-2-yl]methylene-2-thioxodihydro-4,6(1H,5H)-pyrimidinedione in brain tumor stem cells in vitro ( A cell of brain tumor stem cells comprising the step of treating a 5-[1-(4-hydroxyphenyl)-1H-pyrrol-2-yl]methylene -2-thioxodihydro-4,6(1H,5H)-pyrimidinedione) compound. How to induce death.

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