JP2019026611A - Tumor treatment composition - Google Patents

Abstract

To provide a novel tumor treatment composition.SOLUTION: In each compound library, a compound having a proliferation inhibitory effect on a glioma stem cell strain is identified, to provide a tumor treatment composition containing the compound.SELECTED DRAWING: None

Description

  The present invention relates to a tumor treatment composition characterized by combining fluoropalmitic acid as an active ingredient with temozolomide as an active ingredient. The present invention also relates to a tumor treatment composition comprising fluoropalmitic acid as an active ingredient.

(Brain tumor)
Brain tumors (intracranial tumors) are a generic term that includes not only neoplasms that develop within the skull but also occupying lesions such as hamartomas and granulomas.
In general, (1) gliomas arising from neuroepithelial tissues, (2) tumors arising from nerve sheath cells, (3) tumors arising from brain membranes and related tissues, (4) malignant lymphomas, (5) blood vessels Primary tumor, (6) germinoma, (7) teratomas-like tumor (craniopharyngioma, etc.) and tumor-like lesion (eg hamartoma), (8) anterior pituitary tumor, (9) tumor of surrounding tissue It is classified into intracranial extension (such as pheochromocytoma and chordoma) and (10) metastatic brain tumor.

(Glioma)
Glioma is a general term for tumors (neoplasms) that develop from cells of cell lines (glial cells, nerve cells, pineal parenchymal cells) that differentiate from the medullary epithelium.
In general, glial cell lineage tumors are astrocytic, oligodendroma, ependymoma, and glioblastoma, and neuronal cell lines are medulloblastoma, medullary epithelioma, neuroblastoma, neuroblastoma Nodular neuromas and neuronal gliomas, and pineal parenchymal cell lines are pineal cell tumor and pineal blastoma.
Of the malignant gliomas, the proportion of anaplastic astrocytoma is about 30%, and the proportion of glioblastoma is about 57%, which accounts for about 80% or more.

(Glioblastoma)
Glioblastoma (glioblastoma) accounts for about 9% of primary brain tumors and is the least differentiated malignant tumor of glioma (WHO grade IV). It occurs frequently in the adult cerebral hemisphere and infiltrates in the brain parenchyma chronically.
The standard treatment method is to remove the tumor as long as it does not cause new neurological dropout symptoms by craniotomy. And radiation therapy and chemotherapy are performed after an excision operation.
However, even with the above treatment, almost 100% of cases have relapsed, with an average survival time of about one year from the onset, and the prognosis is very poor.
Accordingly, there is a very high demand for development of a therapeutic agent for glioblastoma.

In addition, the present inventors have made the following patent applications and non-patent literature publications.
Patent Document 1 discloses that “GSK3β is involved in cancer cell survival / proliferation and provides a new means for cancer treatment / diagnosis”.
Patent Document 2 discloses “a brain tumor treatment method characterized by administration of a compound having a plurality of GSK3β inhibitory effects, particularly a recurrent brain tumor treatment method, and a brain tumor treatment kit capable of performing the treatment method”.
Patent Document 3 discloses “a brain tumor therapeutic composition containing floxuridine and / or pricamycin as an active ingredient”.
Non-Patent Document 1 discloses that “GSK3β is involved in the survival and proliferation of human glioblastoma cells”.

  However, in the above-mentioned patent document and non-patent document, a tumor treatment composition characterized by combining fluoropalmitic acid as an active ingredient with temozolomide as an active ingredient, and a tumor treatment composition containing fluoropalmitic acid as an active ingredient There is no disclosure or suggestion.

No. 2006-0703202 JP 2012-041314 A Japanese Unexamined Patent Publication No. 2016-210718

Miyashita K, et al. Potential therapeutic effect of glycogen synthese kinase 3βinhibition against human glioblastoma. Clin Cancer Res 2009; 15 (3): 887-97.

  As mentioned above, glioblastoma is a glioma with a very low degree of differentiation and the highest malignancy. Actual tumors often cannot be completely removed by surgery, are resistant to existing anticancer drug treatment and radiation treatment, and are extremely refractory. Thus, since there is no effective treatment for this tumor, its life prognosis has not been improved in the last 30 years, and the development of a new therapeutic agent for brain tumors is eagerly desired.

  In order to solve the above problems, the present inventors have identified a compound having a growth inhibitory effect on a cell line related to a brain tumor from a single compound or in combination with temozolomide from each compound library, A brain tumor therapeutic composition comprising a compound is provided.

  The tumor therapeutic composition of the present invention has a growth inhibitory effect on cell lines related to brain tumors.

That is, the present invention is as follows.
1. A composition for treating tumors, comprising combining fluoropalmitic acid as an active ingredient with temozolomide as an active ingredient.
2. 2. The tumor treatment composition according to item 1, wherein the fluoropalmitic acid is 2-fluoropalmitic acid.
3. 3. The tumor treatment composition according to item 1 or 2, wherein the tumor is a brain tumor.
4). 4. The tumor treatment composition according to item 3 above, wherein the brain tumor is glioma.
5). 5. The tumor treatment composition according to item 4, wherein the glioma is glioblastoma.
6). A tumor treatment composition comprising fluoropalmitic acid as an active ingredient.
7). 6. The tumor treatment composition according to item 5 above, wherein the fluoropalmitic acid is 2-fluoropalmitic acid.
8). 8. The tumor treatment composition according to item 6 or 7, wherein the tumor is a brain tumor.

The growth-inhibition result with respect to the glioma stem cell strain by 2-fluoro palmitic acid (2-FPA, candidate drug) and / or temozolomide (TMZ) in Example 1. FIG. The vertical axis shows the cell viability (%) with the cell viability of the drug-free sample as 100%, and the horizontal axis shows the drug concentration (μM). The growth suppression result with respect to the glioma stem cell line by 2-fluoro palmitic acid and / or temozolomide in Example 2. FIG. The vertical axis represents the cell viability (%) with the cell viability of the drug-free sample as 100%, and the horizontal axis represents the drug concentration (μM). A: Results for 528 PN cells. B: Results for 1123M cells. The observation result of the sphere in Example 3. The upper left figure shows the result of DMSO (control), the upper middle figure shows the result of administration of 2-fluoropalmitic acid (2-FPA) 2 μM, and the upper figure on the right shows the result of administration of 2-FPA 20 μM. The lower left figure shows the result of single agent administration of temozolomide (TMZ) 2 μM, the lower middle figure shows the result of joint administration of TMZ 2 μM + 2-FPA 2 μM, and the lower right figure shows the result of combined administration of TMZ 2 μM + 2-FPA 20 μM. The measurement result of the sphere number of 100 micrometers or more in Example 3. FIG. The vertical axis represents the number of spheres of 100 μm or more, and the horizontal axis represents the type of added drug. A: The result of adding 2-fluoropalmitic acid (2-FPA) single agent. “DMSO” is the result of DMSO, “2 μM” is the result of administration of 2-FPA 2 μM, “4 μM” is the result of administration of 2-FPA 4 μM, “8 μM” is the result of administration of 2-FPA 8 μM, “20 μM” indicates the result of single-agent administration of 2-FPA 20 μM. B: The result of adding temozolomide (TMZ) and 2-fluoropalmitic acid in combination. “DMSO” is the result of DMSO, “TMZ (2 μM)” is the result of TMZ 2 μM single administration, “TMZ + 2 μM” is the result of TMZ 2 μM + 2-FPA 2 μM, “TMZ + 4 μM” is the result of TMZ 2 μM + 2-FPA 4 μM, “TMZ + 8 μM” indicates the result of combined administration of TMZ 2 μM + 2-FPA 8 μM, and “TMZ + 20 μM” indicates the result of combined administration of TMZ 2 μM + 2-FPA 20 μM. The measurement result of the light absorbency in Example 4. The vertical axis represents the logarithm of absorbance, and the horizontal axis represents the culture time. “DMSO” indicates the result of DMSO, “TMZ 250 μM” indicates the result of single agent administration of TMZ 250 μM, “1 μM” indicates the result of single agent administration of 2-FPA 1 μM, and “1 μM + TMZ” indicates the result of combined administration of 2-FPA 1 μM + TMZ 250 μM.

(Tumor therapeutic composition of the present invention)
The tumor treatment composition of the present invention contains fluoropalmitic acid and temozolomide, which are active ingredients, or fluoropalmitic acid, which is an active ingredient.

{Temozolomide (TMZ)}
Oral applied to malignant glioma with chemical name 3,4-dihydro-3-methyl-4-oxoimidazo [5,1-d] -as-tetrazine-8-carboxamide, molecular formula C ₆ H ₆ N ₆ O ₂ It is an anticancer drug that can be administered. Temozolomide binds an atomic group called an alkyl group to a part of nucleic acid such as DNA of cancer cells, thereby inhibiting DNA synthesis and killing cancer cells.
Glioblastoma has the problem of acquiring resistance to temozolomide in the course of treatment with oral temozolomide. One cause of resistance to temozolomide is the presence of glioma stem cells with tumorigenicity and treatment resistance.
It is also marketed under the trade name “Temodar”.

{Fluoropalmitic acid}
Fluoropalmitic acid includes all compounds represented by the molecular formula C ₁₆ H ₃₁ FO ₂ and their derivatives. Examples of the fluoropalmitic acid include 2-fluoropalmitic acid (2-FPA), 3-fluoropalmitic acid, 4-fluoropalmitic acid, 16-fluoropalmitic acid, and derivatives thereof. Preferably, it is 2-fluoro palmitic acid.
2-Fluoropalmitic acid is a sphingosine synthesis inhibitor.

(Pharmaceutically acceptable salt)
Not only the above-mentioned compounds but also pharmaceutically acceptable salts of the above-mentioned compounds, such as basic acid addition salts, inorganic or organic acids or alkali metal salts, alkaline earth metal salts and the like, also relate to the present invention. It can be used as a compound having a growth inhibitory effect on glioma stem cells (glioma cells).
In addition, the above-mentioned compounds are usually known pharmacologically acceptable carriers, excipients, diluents, extenders, disintegrating agents, stabilizers, preservatives, buffers, emulsifiers, fragrances, Coloring agents, sweeteners, thickeners, flavoring agents, solubilizers, other additives, specifically water, vegetable oils, alcohols such as ethanol or benzyl alcohol, polyethylene glycol, glycerol triazetate gelatin, lactose, Oral or parenteral administration in the form of tablets, capsules, elixirs, microcapsules, injections, solutions, suspensions, etc., mixed with carbohydrates such as starch, magnesium stearate, talc, petrolatum etc. can do.
The administration route of the active ingredient used in the tumor therapeutic composition of the present invention is not particularly limited, and may be, for example, oral administration, intravenous injection, subcutaneous injection, intramuscular injection, local injection, intraperitoneal administration, and the like. Intravenous injection or oral administration is preferred.

(Tumor treatment kit of the present invention)
In the tumor treatment kit of the present invention, each of the active ingredients is separately contained in a plurality of containers, or each effective ingredient is used so that fluoropalmitic acid and temozolomide can be administered simultaneously, separately, or sequentially. The components are stored in a single container having a plurality of spatially separable regions.
Furthermore, the tumor treatment kit of the present invention may include instructions regarding administration of each active ingredient. In addition, this instruction | indication describes the administration schedule of each active ingredient in the tumor treatment method of this invention shown below.
In particular, each active ingredient in the tumor treatment kit of the present invention is preferably an orally administrable tablet, but can be a sustained-release granule, syrup, fine granule, and / or injection.

(Tumor treatment method of the present invention)
In the tumor treatment method of the present invention, the dose of fluoropalmitic acid and temozolomide, the frequency of administration and the administration interval are not particularly limited, and the purpose of prevention (particularly prevention of recurrence) and / or clinical treatment, the type of disease, the weight of the patient It is appropriately selected according to conditions such as age and severity of disease.
For example, the following dosage forms can be used, but are not particularly limited.
Temozolomide: 10 mg / m ² body surface area to 500 mg / m ² body surface area, preferably 50 mg / m ² body surface area to 100 mg / m ² body surface area, more preferably 60 mg / m ² body surface area to 90 mg / m ² body surface area, most preferably 65 mg / m ² body surface area to 85 mg / m ² body surface area is administered once a day or several times a day (morning, noon, evening). In addition, 10 mg / m ² body surface area to 500 mg / m ² body surface area, preferably 50 mg / m ² body surface area to 300 mg / m ² body surface area, more preferably 100 mg / m ² body surface area to 250 mg / m ² body surface area, most preferably 150 mg / m ² body surface area to 200 mg / m ² body surface area can be administered continuously for 3-7 days in 3-5 weeks, preferably only 5 days in 4 weeks.
Fluoropalmitic acid when administered in combination with temozolomide: 0.010 mg / kg body weight to 0.300 mg / kg body weight, 0.020 mg / kg body weight to 0.250 mg / kg body weight or 0.011 mg / kg body weight to 0.212 mg / kg body weight Preferably, 0.044 mg / kg body weight to 0.212 mg / kg body weight, more preferably 0.044 mg / kg body weight to 0.088 mg / kg body weight is divided into once a day or several times a day (morning, noon, evening) To administer.
Fluoropalmitic acid when administered as a single agent: 0.010 mg / kg body weight to 0.300 mg / kg body weight, 0.011 mg / kg body weight to 0.212 mg / kg body weight, 0.044 mg / kg body weight to 0.088 mg / kg body weight or 0.050 mg / kg The body weight may be 0.250 mg / kg body weight, preferably 0.044 mg / kg body weight to 0.212 mg / kg body weight, more preferably 0.088 mg / kg body weight to 0.212 mg / kg body weight once a day or the number of days Dosing in divided doses (morning, noon, evening).
In addition, it can be administered every day, once every few days, or several times every other week. It can also be administered continuously for 3-7 days in 3-5 weeks, preferably only 5 days in 4 weeks.

(Combination administration of temozolomide and fluoropalmitic acid)
Temozolomide and fluoropalmitic acid are administered simultaneously, administered several hours apart, administered one day apart, administered several days apart, administered several weeks apart, Administration may be performed at intervals of several months, but simultaneous administration is preferred. More specifically, simultaneous administration once a day for five consecutive days once a month is preferable.
Alternatively, temozolomide and fluoropalmitic acid may be mixed with a physiologically acceptable solvent, carrier, excipient, binder, etc. to form a pharmaceutical composition, and then the composition may be administered to a patient.
Although it does not specifically limit as a solvent, For example, water, alcohol, physiological saline, etc. are mentioned.
Although it does not specifically limit as a support | carrier, For example, an acid, a base, a buffer solution, a stabilizer, an isotonizing agent, a preservative, etc. are mentioned.
Although it does not specifically limit as an excipient | filler, For example, a filler, a coagulant, a slipping agent, a disintegrating agent, a pigment | dye, a sweetener, a fragrance | flavor, a coating agent etc. are mentioned.
The binder is not particularly limited. For example, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, polyvinyl Examples include pyrrolidone and the like.
Furthermore, when temozolomide and fluoropalmitic acid are formulated separately, the separately formulated products may be mixed using a diluent or the like at the time of use to form a mixture, and then the mixture may be administered to the patient.

(Treatment subject of the tumor treatment composition, tumor treatment kit or tumor treatment method of the present invention)
The treatment target of the tumor treatment composition, tumor treatment kit or tumor treatment method of the present invention includes all malignant tumors (or possible malignant tumors) at any stage (early, advanced, end stage). For example, adrenal tumor, breast cancer, cervical cancer, ovarian cancer, colon cancer, endometrial cancer, esophageal cancer, renal cancer, liver cancer, lung cancer, lymphoma, leukemia, testicular cancer, pancreatic cancer, prostate cancer, stomach cancer, thyroid cancer, bladder Includes cancer and brain tumors.
Preferably brain tumor {glioma arising from neuroepithelial tissue, tumor arising from nerve sheath cells, tumor arising from brain membrane and related tissues, malignant lymphoma, angiogenic tumor, germinoma, teratomatous tumor (cranium Laryngeal tumors) and tumor-like lesions (such as hamartomas), anterior pituitary tumors, intracranial spread of tumors in surrounding tissues (such as pheochromocytoma and chordoma) and metastatic brain tumors}, especially gliomas {glial cells (Astrocytoma, oligodendroma, ependymoma and glioblastoma), neurons (medulloblastoma, medullary epithelioma, neuroblastoma, ganglionous neuroma and neuroglioma) , Pineal parenchymal cells (pineocytoma and pineoblastoma)} and glioblastoma.
Furthermore, tumors (primary lesions) in which these tumors recur or remain after craniotomy are also treated.

  As used herein, the term “patient” refers to a mammal, preferably a dog, cat, horse, more preferably a human.

The tumor treatment composition, tumor treatment kit, and / or tumor treatment method of the present invention provide for reducing the size of cancer tissue and / or tumor, delaying the growth of tumor cells, and / or eliminating them.
Reduction in tumor size and / or inhibition of growth can be measured by response criteria (RECIST) guidelines in solid tumors.

EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated in detail, the scope of the present invention is not limited by these Examples.
The following examples have been carried out with the approval of the Kanazawa University Medical Ethics Committee.

(Pre-screening from drug library)
The existing 1301 compounds were selected as candidate drugs from five drug libraries including FDA Approved Drug Library, Kinase Inhibitor Library, Phosphatase Inhhibitor Library, Fatty acid Drug Library, and ICCB Known Bioactives Library. And the compound which has the growth inhibitory effect with respect to a glioma stem cell was selected.
Details are as follows.

(Pre-screening by WST-8 proliferation assay)
In order to examine the growth inhibitory effect of glioma stem cells using the glioma stem cell line, WST-8 proliferation assay was performed according to the method shown below. 1123M (Mesenchymal type) was used as the glioma stem cell line. As the WST-8 reagent used in the WST-8 proliferation assay, Microbial Viability Assay Kit-WST (DOJINDO) was used. A multifunctional tabletop pipette station: EDR-384SII (BioTec) was used for the addition of various reagents.
(1) Glioma stem cells were added to DMEM / F-12 medium (containing B27, EGF, bFGF, Glutamax, and Penicillin-Streptavidin) and cultured. Furthermore, the cell suspension was prepared by diluting with DMEM / F-12 to 2000 cells / well.
(2) Temozolomide (TMZ, manufactured by SIGMA-ALDRICH) was added to the cell suspension of (1) at 50 μM to prepare a “TMZ single agent” sample.
(3) For 1301 candidate drugs, each drug was added to the fine cell suspension of (1) at three different concentrations of 1 μM, 5 μM, and 20 μM to form a “candidate drug single agent” sample.
(4) 50 μM temozolomide and 1 μM, 5 μM, and 20 μM candidate drugs were added to the cell suspension of (1) to prepare a combined sample of “TMZ and candidate drug”.
(5) 36 μl of the cell suspension (1) to (4) was seeded in each well of the non-contact 384-well plate. 4 μl of coloring reagent was added to each well. The cells were cultured at 37 ° C. for 3 hours in an incubator. After culturing, the absorbance at 450 nm was measured using a microplate reader {Readers Infiniten 200 Pro (TECAN)}. From the measurement results, the cell viability was calculated with the control without addition of the drug as 100%.
(6) The cell survival rate by the “additive effect” was calculated from the calculation results of the cell survival rates of “TMZ single agent” and “candidate drug single agent”. In more detail, it computed with the following formulas.
Cell survival rate (%) by “additive effect” = 100− (100−cell survival rate of “temozolomide single agent”) − (100−cell survival rate of “candidate drug single agent”)
(7) The cell viability calculation result of “TMZ and candidate drug” in (4) is 5% or more lower than the cell viability due to “additive effect” in (6), that is, “additive effect” Candidate drugs that showed a “synergistic effect” exceeding that were extracted.

(result)
54 candidate drugs were extracted from 1301 compounds by prescreening.
As a representative example, the result of 2-fluoropalmitic acid is shown in FIG.
As described above, 54 candidate drugs including 2-fluoropalmitic acid (2-FPA) were extracted. These drugs were confirmed to have a synergistic effect, not just an additive effect, when used in combination with temozolomide.

(Screening by WST-8 proliferation assay at low concentration)
The 54 candidate drugs extracted in Example 1 were subjected to WST-8 proliferation assay under lower concentration conditions. Two subtypes of glioma stem cell lines 1123M (Mesenchymal type) and 528PN (Proneural type) were used. Details are as follows.
(1) Glioma stem cells were added to DMEM / F-12 medium and cultured. Furthermore, the cell suspension was prepared by diluting with DMEM / F-12 to 2000 cells / well.
(2) Temozolomide was added to the cell suspension of (1) at 50 μM to prepare a “TMZ single agent” sample.
(3) For 54 candidate drugs, each drug was added to the fine cell suspension of (1) at three different concentrations of 0.1 μM, 0.5 μM, and 1 μM to form a “candidate drug single agent” sample.
(4) 50 μM temozolomide and 0.1 μM, 0.5 μM, and 1 μM candidate drugs were added to the cell suspension of (1) to prepare a combined sample of “TMZ and candidate drug”.
(5) 200 μl of the cell suspension (1) to (4) was seeded in each well of a non-contact 96-well plate. 20 μl of coloring reagent was added to each well. The cells were cultured at 37 ° C. for 3 hours in an incubator. After culturing, the absorbance at 450 nm was measured using a microplate reader. From the measurement results, the cell viability was calculated with the control without addition of the drug as 100%.
(6) One drug with the lowest cell viability was selected for “Candidate drug alone” in (3) and “TMZ and candidate drug” in (4).

(result)
2-Fluoropalmitic acid was extracted from 54 candidate drugs by screening. The result of 2-fluoropalmitic acid is shown in FIG.
It was confirmed that 2-fluoropalmitic acid has a growth inhibitory effect (cell viability reduction effect) on glioma stem cells (1123M and 528PN) in the cases of “single agent” and “combination with TMZ”.

(Sphere formation assay)
Human brain tumor stem cells established from human brain tumors are transplanted into mice, and glioma stem cell line subtype KGS-01 established from the formed tumors is used to produce temozolomide (TMZ) and 2-fluoropalmitic acid (2-FPA, In order to quantify the growth inhibitory effect of CAYMAN CHEMICAL COMPANY) on glioma stem cells, a sphere formation assay was performed by the following method.
(1) KGS-01 cells were suspended in DMEM / F-12 medium (containing B21, EGF, bFGF, Glutamax, and Penicillin-Streptavidin) and seeded in a 96-well plate at a concentration of about 1000 cells / well.
(2) 2-FPA 2 μM, 2-FPA 4 μM, 2-FPA 8 μM, 2-FPA 20 μM, TMZ 2 μM, TMZ 2 μM + 2-FPA 2 μM, TMZ 2 μM + 2-FPA 4 μM, TMZ 2 μM + 2-FPA 8 μM, or TMZ 2 μM + 2-FPA 20 μM were added to each well. Since the drug solvent was DMSO and the drug dose was adjusted to 1 μl, 1 μl DMSO was added to the control wells.
(3) The cells were cultured at 37 ° C. for 10 days in an incubator.
(4) After culturing, the size and number of spheres in each well were observed, and the number of spheres of 100 μm (micrometer) or more was counted.

(result)
A representative example of the observation results is shown in FIG.
In the temozolomide single agent and the 2-fluoropalmitic acid single agent, the number of spheres formed was smaller than that in DMSO. Furthermore, the combined use of temozolomide and 2-fluoropalmitic acid reduced the number of spheres formed, and the sphere size also decreased. From this, the stem cell nature of glioma stem cells was inhibited by the combined use of temozolomide and 2-fluoropalmitic acid.
The measurement result of the number of spheres of 100 μm or more is shown in FIG.
In 2-fluoropalmitic acid single agent, the number of spheres of 100 μm or more was lower than DMSO and temozolomide single agent.
In addition, 2-fluoropalmitic acid was more effective than temozolomide alone and 2-fluoropalmitate alone when used in combination with temozolomide.
From the above, it has been quantitatively confirmed that 2-fluoropalmitic acid has an effect of enhancing the action of temozolomide on proliferation of glioma stem cells. Moreover, it confirmed that the stem cell nature of a glioma stem cell could be inhibited by combined use of temozolomide and 2-fluoro palmitic acid.

(AlamarBlue proliferation assay)
Using the glioma cell lines U87, U251, T98G, SNB19 and A172, AlamarBlue proliferation assay was performed by the following method.
(1) Each cell was suspended in DMEM (1X) medium (FBS: Fetal Bovine Serum added to 0.1%) and seeded in a 96-well plate at a concentration of about 1000 cells / well.
(2) The drug was added to each well so that DMSO 1 μl, TMZ 250 μM, 2-FPA 1 μM, TMZ 250 μM + 2-FPA 1 μM.
(3) The cells were cultured at 37 ° C. for 96 hours in an incubator, and the absorbance (absorption wavelength 570 nm) was measured at 0 hours, 24 hours, 48 hours, 72 hours, and 96 hours after the start of the culture.

The results are shown in FIG.
In all of the five types of human brain tumor-derived cell lines used, it was confirmed that the combined use of 2-fluoropalmitic acid and temozolomide had lower absorbance than temozolomide alone and further suppressed cell proliferation.
The five types of cell lines are different from each other in the excised patients, differ morphologically and molecularly, and have different growth rates. For example, O-6-methylguanine-DNA methyltransferase (MGMT) subtypes are all methylated (methylated) except for T98 cells, and T98 is a mixture of methylated and unmethylated (unmethylated). Therefore, the cell growth inhibitory effect by the combined use of 2-fluoropalmitic acid and temozolomide was not limited by the morphological and molecular biological aspects of the brain tumor, and was confirmed to be effective for all brain tumors.
Therefore, it was confirmed that 2-fluoropalmitic acid has an effect of enhancing the action of temozolomide even at a low concentration of 1 μM.

(General)
This example confirmed that fluoropalmitic acid has an effect of enhancing the growth inhibitory action of temozolomide on glioma stem cells. Furthermore, it was confirmed that the effect was not a simple additive effect but a synergistic effect.
In addition, it was confirmed that fluoropalmitic acid alone has a growth inhibitory effect on glioma stem cells.
Fluoropalmitic acid has a therapeutic effect on glioma, particularly glioblastoma, by having a growth inhibitory effect on glioma stem cells.
In addition, fluoropalmitic acid has an effect of inhibiting stem cell properties of glioma stem cells when used in combination with temozolomide.

  In the present invention, a tumor therapeutic composition can be provided.

Claims (8)

A composition for treating tumors, comprising combining fluoropalmitic acid as an active ingredient with temozolomide as an active ingredient.
The tumor treatment composition according to claim 1, wherein the fluoropalmitic acid is 2-fluoropalmitic acid.
The tumor treatment composition according to claim 1 or 2, wherein the tumor is a brain tumor.
The tumor treatment composition according to claim 3, wherein the brain tumor is glioma.
The tumor treatment composition according to claim 4, wherein the glioma is glioblastoma.
A tumor treatment composition comprising fluoropalmitic acid as an active ingredient.
The tumor treatment composition according to claim 5, wherein the fluoropalmitic acid is 2-fluoropalmitic acid.
  The tumor treatment composition according to claim 6 or 7, wherein the tumor is a brain tumor.