WO2022131667A1 - Composition pharmaceutique pour améliorer l'effet de traitement de mélanome, comprenant un inhibiteur du facteur 2 de transcription des oligodendrocytes en tant que principe actif - Google Patents

Composition pharmaceutique pour améliorer l'effet de traitement de mélanome, comprenant un inhibiteur du facteur 2 de transcription des oligodendrocytes en tant que principe actif Download PDF

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WO2022131667A1
WO2022131667A1 PCT/KR2021/018584 KR2021018584W WO2022131667A1 WO 2022131667 A1 WO2022131667 A1 WO 2022131667A1 KR 2021018584 W KR2021018584 W KR 2021018584W WO 2022131667 A1 WO2022131667 A1 WO 2022131667A1
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transcription factor
olig2
inhibitor
pharmaceutical composition
melanoma
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PCT/KR2021/018584
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Korean (ko)
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황재성
이지은
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경희대학교 산학협력단
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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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a pharmaceutical composition for enhancing the therapeutic effect of melanoma, and more particularly, a composition for enhancing the melanoma therapeutic effect of dabrafenib comprising an inhibitor of expression or activity of the transcription factor Olig2 (Oligodendrocyte transcription factor 2) is about
  • Skin cancer is a malignant tumor of the skin, and representative types include basal cell carcinoma, squamous cell carcinoma, and melanoma. Among them, melanoma can occur in any area where melanocytes exist, such as the skin, eyeballs, and cerebrospinal fluid membranes. known as a tumor.
  • melanoma The main site of metastasis of melanoma is the skin other than the primary lesion, but any other organs such as lymph nodes, bones, lungs, liver, and spleen can also be involved.
  • melanoma has high resistance to conventional chemotherapeutic agents and radiation, and therefore is an intractable disease for which there is no special treatment method to date when abnormally advanced or recurrent.
  • Transcription factor Olig2 is a transcription factor expressed in the central nervous system during embryonic development and is known to play a role in regulating the differentiation and cell cycle progression of oligodendrocytes (oligodendrocytes) and promoting the growth of brain tumors. Transcription factor Olig2 was confirmed to be overexpressed in cells other than oligodendroglioma (oligodendroglioma), such as lung cancer, breast cancer, and melanoma (PCT application number: 10-2017-7027034). The effect on the efficacy of therapeutic agents has not yet been specifically studied.
  • the present inventors have made research efforts to discover substances for treating melanoma.
  • dabrafenib When treated in combination with dabrafenib, it was confirmed that the growth of melanoma cells was more effectively inhibited than when dabrafenib alone was treated.
  • the present invention was completed by confirming the fact that the recurrence of melanoma patients administered with brafenib can also be suppressed.
  • an object of the present invention is to provide a pharmaceutical composition for preventing or treating melanoma, comprising an inhibitor of BRAF inhibitor and Olig2 (Oligodendrocyte transcription factor 2) gene or protein expression or activity.
  • Olig2 Oledendrocyte transcription factor 2
  • the present invention provides a pharmaceutical composition for enhancing the effect of preventing or treating melanoma used in combination with a BRAF inhibitor, wherein the composition comprises an inhibitor of the expression or activity of transcription factor Olig2 (Oligendrocyte transcription factor 2) as an active ingredient.
  • Olig2 Oligendrocyte transcription factor 2
  • the present invention includes a BRAF inhibitor and an Olig2 (Oligodendrocyte transcription factor 2) gene or inhibitor of expression or activity of a gene or protein, preventing or treating melanoma; Or it provides a pharmaceutical composition for preventing or treating metastasis of melanoma.
  • Olig2 Oligodethelial growth factor 2
  • the BRAF inhibitor may be dabrafenib.
  • the present invention provides a method for preventing or treating melanoma in combination with a BRAF inhibitor; Or as a pharmaceutical composition for preventing or enhancing the effect of metastasis of melanoma, the composition provides a pharmaceutical composition comprising an inhibitor of expression or activity of transcription factor Olig2 (Oligendrocyte transcription factor 2) as an active ingredient .
  • Olig2 Oligendrocyte transcription factor 2
  • the BRAF inhibitor may be dabrafenib.
  • the transcription factor Olig2 (Oligodendrocyte transcription factor 2) inhibitor is an antisense oligonucleotide specific for the transcription factor Olig2 (Oligodendrocyte transcription factor 2), small interfering RNA (siRNA), short hairpin It may be any one selected from the group consisting of RNA (short hairpin RNA; shRNA) and ribozyme.
  • the transcription factor Olig2 (Oligodendrocyte transcription factor 2) protein activity inhibitor is a transcription factor Olig2 (Oligodendrocyte transcription factor 2) protein that specifically binds to a protein, peptide, peptide mimetics, It may be any one selected from the group consisting of aptamers, antibodies, and natural products.
  • the combination may be administered simultaneously (simultaneous), separately (separate) or sequentially (sequential).
  • the Olig2 expression inhibitor when administered in combination with dabrafenib, it is confirmed that the melanoma treatment effect of dabrafenib is enhanced. is expected to be useful.
  • Figure 2a is the result of confirming the cell proliferation rate in A375 cells in which the expression of Olig2 is suppressed.
  • Figure 2b is the result of confirming the cell proliferation rate in 501mel cells in which the expression of Olig2 is suppressed.
  • Figure 2c is a result of confirming the number of cells through cell count analysis in A375 cells in which Olig2 expression is suppressed.
  • Figure 2d is a result of confirming the number of cells through cell count analysis in 501mel cells in which the expression of Olig2 is suppressed.
  • 3a is a result of microscopic observation of cell migration over 24 hours and 48 hours in A375 cells in which Olig2 expression is suppressed.
  • Figure 3b is the result of quantification of the difference in the area of the wound gap according to the lapse of 24 hours and 48 hours in A375 cells in which the expression of Olig2 is suppressed compared to the control group.
  • Figure 4a is the result of observing the cell migration according to the lapse of 24 hours and 48 hours in 501mel cells in which the expression of Olig2 is suppressed under a microscope.
  • Figure 4b is the result of quantification of the difference in the area of the wound gap according to the lapse of 24 hours and 48 hours in 501mel cells in which Olig2 expression is suppressed compared to the control group.
  • 5a is a result of confirming cell mobility using a transwell, and is a result of staining A375 cells that migrated through the pores of the chamber with DAPI and observing it under a microscope.
  • Figure 5b is the result of counting and quantifying the nuclei stained by using the Image J program of the migrated A375 cells.
  • 6a is a result of confirming cell invasiveness using a transwell, and is a result of staining A375 cells infiltrating through Matrigel instead of an extracellular matrix with DAPI and observing it under a microscope.
  • Figure 6b is the result of counting and quantifying the stained nucleus of the infiltrating A375 cells using the Image J program.
  • the present inventors used an inhibitor of expression or activity of the transcription factor Olig2 (Oligodendrocyte transcription factor 2) and a targeted therapeutic agent, dabrafenib.
  • Olig2 the transcription factor 2
  • dabrafenib a targeted therapeutic agent
  • the present invention provides a pharmaceutical composition for preventing or treating melanoma, comprising an inhibitor of BRAF and Olig2 (Oligodendrocyte transcription factor 2) gene or protein expression or activity.
  • the term "BRAF inhibitor” may be dabrafenib, a targeted therapeutic agent used for the treatment of late-stage melanoma, and in the present invention, dabrafenib is known to regulate cell growth.
  • dabrafenib By suppressing the expression of BRAF, a gene encoding the B-Raf protein, it is being used to treat melanoma and metastatic non-small cell lung cancer.
  • the present invention provides a pharmaceutical composition for enhancing the effect of preventing or treating melanoma used in combination with a BRAF inhibitor, the composition comprising an inhibitor of expression or activity of transcription factor Olig2 (Oligendrocyte transcription factor 2) as an active ingredient It provides a pharmaceutical composition, characterized in that.
  • Olig2 Oligendrocyte transcription factor 2
  • protein may be used synonymously with “polypeptide” or “polypeptide”, and refers to a polymer of amino acid residues. Proteins apply to amino acid polymers in which one or more amino acid residues are artificial chemical mimics of the corresponding naturally occurring amino acids, as well as naturally occurring amino acid polymers, those containing modified residues, and non-naturally occurring amino acid polymers.
  • the transcription factor Olig2 (Oligodendrocyte transcription factor 2, hereinafter Olig2) gene or protein expression or activity inhibitor refers to a substance that inhibits or interferes with the function of Olig2. Such inhibition or interference may be achieved by interfering with the transcription of the Olig2 gene or by inhibiting the translation of the mRNA of the Olig2 gene. Alternatively, the activity of the Olig2 protein may be reduced or inactivated by specifically binding to the active site of the Olig2 protein or by causing a protein structural modification.
  • transcription factor Olig2 (Oligodendrocyte transcription factor 2) gene or protein expression inhibitor) is not limited thereto, but transcription factor Olig2 (Oligendrocyte transcription factor 2 mRNA specific antisense oligonucleotide, small interfering RNA) (small interfering RNA; siRNA), short hairpin RNA (shRNA), and ribozyme may be any one selected from the group consisting of.
  • transcription factor Olig2 Oligendrocyte transcription factor 2 mRNA specific antisense oligonucleotide, small interfering RNA) (small interfering RNA; siRNA), short hairpin RNA (shRNA), and ribozyme may be any one selected from the group consisting of.
  • the inhibitor of the activity may refer to a substance that makes the same type of activity lower than the intrinsic activity that the non-genetically engineered parent cell (eg, wild-type) does not have or has.
  • the term "inhibitor of transcription factor Olig2 (Oligodendrocyte transcription factor 2) protein activity" of the present invention is not limited thereto, a compound, peptide, or peptide mimetic that specifically binds to transcription factor Olig2 (Oligodendrocyte transcription factor 2) protein It may be any one selected from the group consisting of s, aptamers, antibodies, and natural products.
  • nucleotides are the building blocks of oligonucleotides and polynucleotides, and for purposes of the present invention include both naturally occurring and non-naturally occurring nucleotides.
  • nucleotides such as DNA and RNA nucleotides, contain a ribose sugar moiety, a nucleobase moiety and one or more phosphate groups (absent in nucleosides). Nucleosides and nucleotides may also be referred to interchangeably as “unit” or “monomer.”
  • Watson-Crick base pairs are guanine (G)-cytosine (C) and adenine (A)-thymine (T)/uracil (U).
  • Oligonucleotides may include nucleosides with modified nucleobases. For example, 5-methyl cytosine is often used instead of cytosine.
  • complementarily binding may include Watson-Crick base-binding between unmodified nucleobases and modified nucleobases (see, e.g., Hirao et al (2012) Accounts of Chemical Research vol 45 page 2055 and Bergstrom (2009) Current Protocols in Nucleic Acid Chemistry Suppl. 37 1.4.1).
  • the antisense nucleotide is defined as an oligonucleotide capable of regulating the expression of a target gene by hybridizing to a target nucleic acid, in particular, to an adjacent sequence on the target nucleic acid.
  • Antisense oligonucleotides are not inherently double-stranded and therefore not siRNA or shRNA.
  • the small interfering RNA refers to RNA capable of inducing RNAi (RNA interference) that inhibits gene activity.
  • the small interfering RNA may be a miRNA or siRNA capable of inhibiting the expression of the Olig2 gene, and the small interfering RNA may have any form as long as it inhibits the expression or activity of the Olig2 gene.
  • siRNA obtained by chemical synthesis, biochemical synthesis, or in vivo synthesis, or double-stranded RNA of 10 bases or more in which double-stranded RNA of about 40 bases or more is degraded in the body, etc. can be used.
  • the "aptamer” refers to a nucleic acid molecule having binding activity to a predetermined target molecule.
  • the aptamer may be RNA, DNA, modified nucleic acid, or a mixture thereof, and may be in a linear or cyclic form.
  • the shorter the nucleotide sequence constituting the aptamer the shorter the chemical synthesis and mass production. It is known that it is easier, has excellent cost advantages, is easy to chemically modify, has excellent in vivo stability, and has low toxicity.
  • the “antibody” refers to a proteinaceous molecule capable of specifically binding to an antigenic site of a protein or peptide molecule. Such an antibody is a marker gene by cloning each gene into an expression vector according to a conventional method.
  • the protein encoded by The form of the antibody is not particularly limited, and any part thereof is included in the antibody of the present invention as long as it has polyclonal antibody, monoclonal antibody or antigen-binding property, and all immunoglobulin antibodies may be included as well as humanized antibody, etc. It may also contain special antibodies of
  • the antibody includes functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains.
  • a functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and may be Fab, F(ab'), F(ab')2 and Fv.
  • the term "combination” refers to co-administration, but is not limited thereto, but the Olig2 inhibitor of the present invention and dabrafenib are administered in combination simultaneously (simultaneous), separately (separate) or sequentially (sequential). it could be
  • the present inventors confirmed that the preventive or therapeutic effect of melanoma was improved when the oilg2 gene or protein expression or activity inhibitor and dabrafenib were administered in combination through specific examples.
  • melanoma growth can be inhibited by enhancing the effect of dabrafenib when treating melanoma cells in which the expression of Olig2 is suppressed, dabrafenib, a melanoma therapeutic agent. (see Example 4).
  • the inventors of the present invention specifically confirmed that when an inhibitor capable of inhibiting the expression or activity of Olig2 of the present invention is treated, metastasis and growth of melanoma can be significantly inhibited.
  • the inhibitor and dabrafenib were administered in combination, it was confirmed that even a small amount could enhance the melanoma inhibitory effect of dabrafenib.
  • the Olig2 inhibitor can be widely used in the field of melanoma treatment.
  • the pharmaceutical composition may include one or more pharmaceutically acceptable carriers in addition to the active ingredients described above.
  • the pharmaceutically acceptable carrier may be used in a mixture of saline, sterile water, Ringer's solution, buffered saline, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, and one or more of these components, and, if necessary, an antioxidant , buffers, bacteriostatic agents, and other conventional additives may be added.
  • diluents, dispersants, surfactants, binders and lubricants may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets.
  • the pharmaceutical composition may be administered in combination parenterally, subcutaneously, intravenously, intramuscularly, intraperitoneally, intradermally or orally.
  • the drug may be administered orally or directly to a target organ, and the dosage may vary depending on the patient's age, sex, health condition and weight, type of concurrent treatment, administration time, administration method, excretion rate, disease severity, and frequency of treatment. and the nature of the desired effect.
  • it may be administered systemically or locally in consideration of conditions such as the disease to be treated, the need for site-specific treatment, and the amount of the drug to be administered.
  • the pharmaceutical composition may have any pharmaceutical formulation.
  • a form suitable for oral administration as a tablet, capsule, pill, powder, sustained release formulation, solution, or suspension in a form suitable for parenteral injection as a sterile solution, suspension or emulsion, or in a form suitable for topical administration as an ointment or cream. It may be in a suitable form or may be in a form suitable for rectal administration as a suppository.
  • the composition may be a tablet, pill, injection, or a combination thereof.
  • the pharmaceutical composition may be in unit dosage form suitable for single administration of precise dosages.
  • the pharmaceutical composition may be used alone or in combination, or in combination with other therapeutic agents or diagnostic agents.
  • the pharmaceutical composition may be administered concurrently with other agents typically prescribed for the intended disease in accordance with generally accepted medical practice.
  • the pharmaceutical composition can generally be used in a mammal, for example, a human in vivo (in vivo).
  • the present invention provides a method for preventing or treating melanoma, comprising administering a BRAF inhibitor and an Olig2 (Oligodendrocyte transcription factor 2) gene or protein expression or activity inhibitor to an individual in need thereof.
  • Olig2 Oledendrocyte transcription factor 2
  • the term “administration” means providing a given composition of the present invention to a subject by any suitable method.
  • the term "subject” refers to a subject in need of treatment for a disease, and more specifically, human or non-human primates, mice, dogs, cats, horses, and cattle. means mammals.
  • Another aspect provides the use of a BRAF inhibitor and an Oligodendrocyte transcription factor 2 (Olig2) gene or inhibitor of expression or activity for the manufacture of a medicament for the prophylaxis or treatment of melanoma.
  • Olig2 Oligodendrocyte transcription factor 2
  • Human melanoma cells, A735, 501mel and HM3KO cells were treated with Dulbecco's modified eagle supplemented with 10% Fetal Bovine Serum (Gibco, Los Angeles, CA, USA) and 1% penicillin/streptomycin.
  • Culture in ⁇ s medium (DMEM; Welgene Inc., Korea).
  • MNT1 a human melanoma cell, was prepared in minimum essential media (MEM; Welgene Inc.) supplemented with 20% FBS, 1% penicillin/streptomycin and 20 mM hydroxyethyl piperazineethanesulfonic acid (HEPES). Korea).
  • NHEMs Normal human melanocytes, NHEMs, were prepared from Human Melanocyte Growth Supplement (HMGS; Invitrogen Life Technologies, Carlsbad, CA, USA) and medium 254 (Medium 254, M-254) containing 1% penicillin/streptomycin. -500; Invitrogen Life Technologies, Carlsbad, CA, USA) in a humidified atmosphere of 37° C. and 5% CO 2 .
  • Ez-Cytox assay kit (Daeil Lab Service Co Ltd, Seoul, Korea) was used. Specifically, 10 ⁇ l of Ez-Cytox solution was added to each well-plate, and then the well-plate was added to 5% Incubated for an additional 1 hour under CO 2 conditions and in an incubator at 37°C. Cell viability was confirmed at 490 nm wavelength by an ELISA reader (Tecan, Mannedorf, Switzerland), and relative cytotoxicity was measured as a percentage of cell viability.
  • Cells were seeded in a 96-well plate at 1 ⁇ 10 4 cells/well, and then transfected using CRISPR/Cas9 plasmid. After 72 hours, the cells were trypsin-treated. 10 ul of trypan blue dye (Sigma Aldrich, St. Louis, MO, USA) was mixed in the same manner as the cell suspension. After staining the cells with trypan blue dye, each sample was repeated three times and the number of cells was counted using a hemocytometer.
  • trypan blue dye Sigma Aldrich, St. Louis, MO, USA
  • RNA total RNA
  • melanoma cells prepared in Example 1-1 were treated with TRIZOL reagent (Takara Bio, Inc., Tokyo, Japan), and then 1/5 of TRIZOL in chloroform (Chloroform, CHCl 3 , 99%, Junsei Chemical, Japan) was treated and mixed.
  • TRIZOL reagent Tekara Bio, Inc., Tokyo, Japan
  • chloroform Chloroform, CHCl 3 , 99%, Junsei Chemical, Japan
  • After mixing the Vortex mixer centrifuged for 15 minutes at 13,000 rpm at a temperature of 4 °C. A certain amount of the transparent supernatant was taken and mixed with the same amount of isopropanol. After inverting the tube centrifuge at 4° C., 13,000 rpm, 20 min. The supernatant was removed and the remaining pellet was diluted with diethyl pyrocarbonate treated water.
  • cDNA complementary DNA
  • RT-PCR reverse transcription polymerase chain reaction
  • melanoma cells were seeded in 12 well plates and then cells were trypsinized 24 h after transfection with CRISPR/Cas9 plasmids.
  • Cells were re-seeded with polycarbonate membranes (Corning Incorporated, Corning, NY, USA) with 8.0 ⁇ m pore size in DMEM medium containing 1% FBS at 5 x 10 4 cells per chamber, followed by 18 h. incubated for a while.
  • the upper chamber was coated with Matrigel® (Corning, Kaiserslautern, Germany) diluted with serum-free DMEM medium, and the bottom of the upper chamber was coated with 1% gelatin.
  • the bottom chamber was filled with DMEM medium containing 10% FBS.
  • DMEM medium containing 10% FBS.
  • Cells that invaded through the Matrigel-coated membrane were fixed in 10% formalin and washed twice with PBS.
  • DAPI 4',6-diamidino-2-phenylindole
  • Invasive cells were detected by fluorescence microscopy at x100 magnification. Stained nuclei were counted and quantified using ImageJ software 1.45s (US National Institutes of Health, Bethesda, MD, USA).
  • CRISPR-Cas9 technique was used in 501mel cells, a malignant melanoma cell, to prepare cells in which the expression of the transfer factor Olig 2 (Oligodendrocyte transcription factor 2) was suppressed.
  • UltraCruz transfection reagent UltraCruz Transfection Reagent (Santacruz Biotechnology, Dallas, TX, USA)
  • 20nt guide sgRNA and Cas9 nuclease encoding three The sc-400670-KO-2 consisting of different gRNA plasmids: Olig2 CRISPR/Cas9 KO plasmids (h2) (Santacruz Biotechnology, Dallas, TX, USA) were transfected.
  • the three gRNAs of the plasmid may be gRNA1: 5'-GGCTGCGTCGTCCACCAAGA-3', gRNA2: 5'-CACCTCGTCGTCTACGTCGT-3' or gRNA3: 5'-CGCCCATAGCCGACACGTGG-3'.
  • cancer cells have a characteristic of infinitely proliferating unlike normal cells
  • the proliferation rate of malignant melanoma cells in which Olig2 expression is suppressed was measured. More specifically, in the malignant melanoma cells (A375 cells and 501mel cells) in which the expression of Olig2 prepared in Example 2 is suppressed, Ez-CyToX cell viability was analyzed according to the concentration and time of treatment with Olig2 gRNA.
  • Metastasis one of the main characteristics of malignant melanoma, is a phenomenon in which cells are separated from the primary tumor and move around. Migrating melanoma cells circulate in blood and lymph vessels, settle in other end organs, form secondary tumors, and eventually metastasize. (NGUYEN, DX, BOS, PD and MASSAGUE, J., 2009. Metastasis: from dissemination to organ-specific colonization. Nature Reviews Cancer, 9 (4), pp. 274.)
  • FIGS. 3a A375 cells
  • 4a 501 mel cells
  • transwell metastasis assay was performed to analyze cell mobility.
  • Cells were seeded into the upper chamber and allowed to migrate through the transwell chamber for 18 hours. Thereafter, the cells that migrated through the pores of the chamber were stained with DAPI and observed under a microscope (see Fig. 5a), and the stained nuclei were counted and quantified using the Image J program.
  • Fig. 5b 18 As time elapsed, it was confirmed that a 63 ⁇ 5.2% decrease in the group treated with 1 ⁇ g of gRNA for Olig2 inhibition and 58 ⁇ 5.0% decreased in the group treated with 2 ⁇ g of gRNA.
  • matrigel was diluted with 0% FBS DMEM medium, and then coated on the upper chamber. was stained with and observed under a microscope (see Fig. 6a), and the stained nuclei were counted and quantified using the Image J program. It was confirmed that the decrease was 48 ⁇ 8.2% in the group treated with 1 ⁇ g, and decreased by 54 ⁇ 8.6% in the group treated with gRNA 2 ⁇ g.
  • the present inventors confirmed that when Olig2 is inhibited through the above results, metastasis of melanoma can be inhibited to a significant level.
  • Example 4 Confirmation of survival rate of malignant melanoma cells when oligodendrocyte transcription factor 2 expression is suppressed by Dabrafenib treatment
  • Dabrafenib a melanoma treatment previously approved by the US FDA as a B-Raf inhibitor
  • a B-Raf inhibitor a melanoma treatment previously approved by the US FDA as a B-Raf inhibitor
  • the cell viability was checked.
  • FIG. 7 and Table 3 below, when dabrafenib was administered to malignant melanoma cells in which Olig2 expression was suppressed, and when dabrafenib was treated to Olig2-expressing melanoma cells It was confirmed that the growth inhibitory effect of melanoma cells was superior.

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Abstract

La présente invention concerne une composition pharmaceutique pour améliorer l'effet de traitement de métastase de mélanome. Plus particulièrement, il a été confirmé que l'effet de prévention ou de traitement de mélanome du dabrafenib a été notablement amélioré lorsque la composition comprenant l'inhibiteur du facteur 2 de transcription des oligodendrocytes (OLIG2) était administrée de manière concomitante avec du dabrafenib. La composition pharmaceutique selon la présente invention permet le traitement efficace d'un mélanome même lorsqu'on utilise moins de dabrafenib qu'une dose existante, et peut donc être exempte du problème de toxicité et d'effets secondaires.
PCT/KR2021/018584 2020-12-18 2021-12-09 Composition pharmaceutique pour améliorer l'effet de traitement de mélanome, comprenant un inhibiteur du facteur 2 de transcription des oligodendrocytes en tant que principe actif WO2022131667A1 (fr)

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KR10-2020-0178294 2020-12-18
KR20200178294 2020-12-18
KR1020210124582A KR20220088294A (ko) 2020-12-18 2021-09-17 Oligodendrocyte transcription factor 2 억제제를 유효성분으로 포함하는 흑색종 치료 효과 증진용 약학적 조성물
KR10-2021-0124582 2021-09-17

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