WO2020071392A1 - Inhibiteur de prolifération de cellules cancéreuses à mauvais pronostic - Google Patents

Inhibiteur de prolifération de cellules cancéreuses à mauvais pronostic

Info

Publication number
WO2020071392A1
WO2020071392A1 PCT/JP2019/038825 JP2019038825W WO2020071392A1 WO 2020071392 A1 WO2020071392 A1 WO 2020071392A1 JP 2019038825 W JP2019038825 W JP 2019038825W WO 2020071392 A1 WO2020071392 A1 WO 2020071392A1
Authority
WO
WIPO (PCT)
Prior art keywords
fibrillarin
cancer
cancer cell
expression
inhibitor
Prior art date
Application number
PCT/JP2019/038825
Other languages
English (en)
Japanese (ja)
Inventor
政一 親泊
Original Assignee
小胞体ストレス研究所株式会社
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 小胞体ストレス研究所株式会社 filed Critical 小胞体ストレス研究所株式会社
Priority to JP2020550471A priority Critical patent/JP7191301B2/ja
Publication of WO2020071392A1 publication Critical patent/WO2020071392A1/fr

Links

Images

Classifications

    • 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
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
    • A61K38/08—Peptides having 5 to 11 amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/12—Drugs for disorders of the urinary system of the kidneys
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the present invention relates to an agent for suppressing the growth of cancer cells having a poor prognosis.
  • effective anti-proliferative agents for various types of cancers with high fibrillarin expression colonal cancer, ovarian cancer, breast cancer, lung adenocarcinoma, glioma, kidney cancer, etc.
  • the present invention relates to a therapeutic agent for a cancer disease to be used. Further, the present invention relates to a method for examining the prognosis of cancer.
  • Fibrillarin is a component of snoRNP (small nucleolar small ribonucleic acid protein) located in the nucleolus in the cell nucleus, and plays a role in rRNA methylation and is transcribed from DNA. It is known to carry out subsequent methylation of the rRNA precursor. And the amino acid sequences of many vertebrate fibrillarins are known and they are widely conserved among vertebrates.
  • human-derived fibrillarin is a protein consisting of 321 amino acids (National Center for Biotechnology Information (NCBI) http://www.ncbi.nlm.nih.gov/), monkey, cow, dog, rat, and rat.
  • a typical compound that suppresses or inhibits the above function of fibrillarin is an anti-fibrillarin antibody. Further, it has been reported that a peptide fragment having no fibrillin activity, which is derived from a fibrillin protein, often acts as a fibrillin activity inhibitor (Patent Document 1). Recently, yamamarin and its derivatives, which are peptides derived from the silkworm, form a complex with fibrillarin (Non-patent Document 1 and Patent Document 2), inhibit the action of fibrillarin, exhibit a cell growth inhibitory effect, and exhibit mitochondrial NADH. It has been reported to inhibit respiration (Non-Patent Documents 2 and 3).
  • the cell growth inhibitory activity caused by the inhibition of fibrillarin action suppresses cell growth by shortening the S phase corresponding to the DNA replication phase and prolonging the quiescent phase, and causes cell proliferation by inducing apoptosis. It does not suppress. That is, it is reported that cell growth is suppressed by controlling the cell cycle (Patent Document 2).
  • Substances that suppress the expression and activity of fibrillarin are agents for suppressing or killing the proliferation of undifferentiated stem cells of ES cells and iPS cells (Patent Document 1), and polycythemia vera caused by the proliferation of erythroid progenitor cells Alternatively, it has been reported that it can be used as a therapeutic agent (Patent Document 2) for essential thrombocythemia caused by proliferation of megakaryocyte precursor cells.
  • fibrillarin even if fibrillarin is controlled, it is considered that it is difficult to control only the cell cycle and to induce the death of cancer cells, etc. Was not considered.
  • the present inventor has found that, in the case of a cancer cell having a high fibrillarin expression level (output), the prognosis of a cancer patient after starting treatment is poor (low survival rate). Therefore, suppression of the fibrillarin expression level (production rate) using fibrillarin siRNA was examined.
  • the present inventor has found that, when the expression level (production amount) of fibrillarin is suppressed using siRNA, the proliferation of cancer cells can be suppressed. Furthermore, by administering fibrillarin siRNA to the transplanted cancer cells, growth suppression of the cancer cells could be achieved.
  • cancer treatment is possible.
  • the present inventors have completed the present invention based on the above findings.
  • the gist of the present invention is as follows.
  • a cancer cell growth inhibitor comprising a fibrillarin expression or activity inhibitor as an active ingredient.
  • the cancer cell growth inhibitor according to [1], wherein the cancer cell is a high fibrillarin-expressing cancer cell.
  • the high fibrillarin-expressing cancer cell is a cancer cell whose fibrillarin RNA expression level is equal to or greater than the median fibrillarin RNA expression level determined from a patient database for each cancer type.
  • a cancer cell growth inhibitor comprising a fibrillarin expression or activity inhibitor as an active ingredient.
  • the cancer cell is any one of a colon cancer cell, an ovarian cancer cell, a breast cancer cell, a lung adenocarcinoma cell, a glioma cancer cell, and a kidney cancer cell, [1] or The cancer cell growth inhibitor according to [2].
  • the cancer cell growth inhibitor according to [4] wherein the cancer cell is a kidney cancer cell.
  • the fibrillarin siRNA comprises an RNA nucleotide sequence (SEQ ID NO: 1: ggucgaggcgggagcucuua) corresponding to the nucleotide sequence at positions 103 to 121 of the human fibrillarin gene, or 1 or 2 in SEQ ID NO: 1
  • the cancer cell growth inhibitor according to [6] wherein the cancer cell growth inhibitor comprises an RNA base sequence having a substituted, inserted, or deleted nucleotide and suppresses human fibrillarin expression.
  • a cancer therapeutic agent comprising the cancer cell growth inhibitor according to any of [1] to [8] as an active ingredient.
  • SEQ ID NO: 1 A fibrillarin siRNA or a modified form thereof comprising a nucleotide sequence of up to 25 nucleotides including the nucleotide sequence of ggucggggcggggcucuua.
  • a method for suppressing cancer cell growth which comprises administering to a cancer patient an inhibitor of the expression or activity of fibrillarin in an amount effective for suppressing cancer cell growth.
  • a method for treating cancer comprising administering to a cancer patient an inhibitor of the expression or activity of fibrillarin in an amount effective for treating cancer.
  • the inhibitor of fibrillarin expression or activity of the present invention can suppress the growth of cancer cells even in malignant cancer cells with high expression of fibrillarin and poor prognosis, and kill cancer cells be able to.
  • the inhibitor of the expression or activity of fibrillarin of the present invention it has become possible to treat cancer diseases even with poor prognosis and low survival rates.
  • FIG. 2 is a diagram showing a test method for suppressing the growth of xenograft renal cell carcinoma by suppressing the fibrillarin expression level (production level).
  • FIG. 3 is a view showing the results of a xenograft renal cell carcinoma growth suppression test by suppressing the fibrillarin expression level (production level).
  • the “inhibitor of fibrillarin expression or activity” of the present invention refers to a drug capable of suppressing the expression of fibrillarin in cancer cells or a drug capable of suppressing or inhibiting the function or activity of fibrillarin.
  • the agent capable of suppressing the expression of fibrillarin that is, the agent capable of suppressing the expression of the fibrillarin gene include antisense, siRNA, and shRNA targeting fibrillarin (for example, human fibrillarin) (above, nucleic acids that target or bind to mRNA) Compounds), miRNAs, decoys, aptamers, nucleic acid compounds such as CpG oligonucleotides, and low molecular weight compounds that regulate the transcription of rRNA such as actinomycin D.
  • the number of bases of the nucleic acid compound such as antisense or siRNA to be used may be 14 to 45, and a suitable number of bases can be used depending on the kind of the nucleic acid compound.
  • the number of bases in antisense is 14 to 30, the number of bases in siRNA is 19 to 25, the number of bases in miRNA is 19 to 25, the number of bases in decoy is 16 to 24, and the number of bases in aptamers is 26 to 45.
  • the number of bases of the CpG oligonucleotide is preferably 16 to 24, respectively.
  • the nucleic acid sequence of the nucleic acid compound for suppressing the expression of fibrillarin is not particularly limited, and an appropriate nucleic acid sequence can be used in accordance with the type of the nucleic acid compound to be used.
  • an appropriate nucleic acid sequence can be used in accordance with the type of the nucleic acid compound to be used.
  • siRNA a human fibrillarin gene sequence consisting of an RNA base sequence corresponding to the base sequence at positions 103 to 121 (SEQ ID NO: 1: ggucggggcgggagcucuua) may be mentioned.
  • those comprising an RNA base sequence of 19 to 25 bases including the base sequence of SEQ ID NO: 1 can also be mentioned.
  • shRNAs containing these nucleotide sequences can also be used, and can be used as siRNAs for suppressing the expression of fibrillarin in cells.
  • examples of the antisense RNA include those having a base sequence of SEQ ID NO: 1 or a base sequence of 19 to 30 bases including the base sequence.
  • Antisense DNAs include those consisting of the nucleotide sequence at positions 103 to 121 (SEQ ID NO: 2: ggtcgaggcgggaggctttta) in the gene sequence of human fibrillarin, or those consisting of a nucleotide sequence of 19 to 30 nucleotides including the nucleotide sequence Is mentioned.
  • the siRNA may be an RNA consisting of an RNA base sequence in which one or two, particularly one base has been substituted, inserted or deleted in SEQ ID NO: 1, or a 19 to 25 RNA base sequence containing the same. Can also be used as long as the expression of fibrillarin is suppressed.
  • the antisense RNA includes an RNA base sequence of SEQ ID NO: 1 in which one to two, particularly one base has been substituted, inserted, or deleted, or a 19 to 30 RNA base sequence including the same. Can be used as long as the expression of fibrillarin is suppressed.
  • the antisense DNA includes a DNA base sequence in which one to two, particularly one base in SEQ ID NO: 2 has been substituted, inserted, or deleted, or a 19 to 30 DNA base sequence including the same. Can also be used as long as the expression of fibrillarin is suppressed. Suppressing the expression of fibularin includes not only stopping expression, but also reducing the expression level.
  • the nucleic acid compound when it is RNA, it may be designed so that it can be produced in vivo.
  • a DNA encoding the RNA can be inserted into an expression vector for mammalian cells.
  • an expression vector include a virus vector and an animal cell expression plasmid.
  • the nucleic acid compound when it is RNA, it may be chemically modified to improve stability.
  • the chemically modified RNA include RNA containing nucleic acid analogs such as phosphorothioate, morpholino phosphorodiamidate, boranophosphate, and LNA (Locked Nucleic Acid), and 2′-O-methylated RNA and 2′-O-. Methoxyethylated RNA and the like can be mentioned.
  • the drug capable of suppressing or inhibiting the function or activity of fibrillin refers to a drug that suppresses or inhibits the function of fibrillin and a drug that suppresses or inhibits the activity of fibrillin.
  • fibrillarin has a function of modifying the methylation of rRNA, and the rRNA is matured by the methylation modification and participates in the formation of liposomes, which are protein factories. Therefore, if the methylation modification of rRNA is inhibited, ribosome formation was not possible, indicating that the function of fibrillarin was inhibited.
  • human rRNA has about 200 methylation modification sites.
  • the introduced methyl group generally has the effect of providing a local hydrophobic environment and weakening hydrogen bonds.
  • 2′-O-methylation of ribose has a role of fixing the twisted structure of ribose to C3′-end type, and contributes to local structure formation of rRNA.
  • the chemical nature of the methylation modification of rRNA plays various roles in the biosynthesis and function of ribosomes (Biochemistry Vol. 85, No. 10, pp. 896-908, 2013).
  • the mechanism of this rRNA methylation modification reaction is as follows. That is, snoRNA having a common sequence called Box C / D forms a base pair with rRNA, and four proteins of fibrillarin, Nop58p, Nop56p, and Snu13p (15.5k in human) (mainly fibrillarin) are formed.
  • a bound Box C / D snoRNP complex is formed, which serves as a guide for determining the methylation site of the target rRNA, and the rRNA is methylated.
  • the donor of the methyl group to be introduced is S-adenosylmethionine (SAM), and methylation modification is performed by the action of methyltransferase. Therefore, an agent that suppresses or inhibits the function of fibrillarin refers to an agent that suppresses or inhibits the above-described mechanism of methylation modification.
  • nucleic acid compounds such as aptamers or peptides can be mentioned.
  • An agent that suppresses or inhibits the activity of fibrillin refers to a molecule that binds to the target of fibrillin but has no activity, and thus suppresses or inhibits the activity of fibrillin, or binds or interacts with fibrillin to inhibit the activity of fibrillin. Or a drug that inhibits.
  • the fibrillin-derived peptide fragment described in Patent Document 1 binds to the fibrillin target but has no fibrillin activity, and thus competes with the fibrillin to inhibit its activity.
  • yamamarin C-terminal amidated peptide (Asp-Ile-Leu-Arg-Gly) -NH 2
  • Non-patent Document 1 and Patent Document 2 N-terminal acylation thereof (particularly, C6 to The 28-acylated derivative binds to fibrillarin and inhibits its activity.
  • Antibodies to fibrillarin also bind to and inhibit its activity.
  • ⁇ The“ cancer cell ”of the present invention refers to a cancer cell highly expressing fibrillarin. Depending on the type of cancer cell, as shown in FIG. 1, the expression level of fibrillarin varies depending on the type of cancer cell. Cells.
  • the median fibrillarin expression level of various cancer cells refers to the median fibrillarin expression level in cancer tissues of at least 50, especially 50 to 150, cancer patients. It may be calculated from data collected in the Cancer Genome Atlas Project by the Human Genome Research Institute.
  • cancer cells as shown in FIGS. 2 to 7, for example, colon cancer, ovarian cancer, breast cancer, lung cancer (especially lung adenocarcinoma), glioma, and kidney cancer have If the expression level of fibrillin in cancer tissue is high, cancer cells are likely to metastasize, resulting in a poor post-operative prognosis and a low survival rate.
  • a cancer cell with high fibrillarin expression can be regarded as a cancer cell of a cancer patient exhibiting a median survival rate or a lower survival rate of various cancer patients.
  • the median survival rate of various cancer patients is the median survival rate of at least 50, especially 50 to 150, cancer patients. What is necessary is just to determine from the data collected by the genome atlas project.
  • the survival rate is defined as, for example, the survival rate after 50 to 3000 days, especially 300 to 1500 days after the start of cancer treatment (including surgical treatment including surgery such as cancer resection, radiation therapy, chemotherapy, etc.). It can be.
  • Survival rates range from 50-150 days (especially 50 days) for colorectal cancer, 50-150 days (especially 50 days) for ovarian cancer, and 50-150 days ( After 100 days), 1000-3000 days (especially 1500 days) for lung cancer (especially lung adenocarcinoma), and 300-1000 days (especially 500 days) for glioma. it can.
  • the inhibitor of the expression or activity of fibrillarin of the present invention is more effective for cancer cells expressing fibrillarin at a high level with poor prognosis.
  • the "cancer therapeutic agent" of the present invention may contain additives and carriers as long as an effective amount of an inhibitor of the expression or activity of fibrillarin exists in such a manner as to exert its function.
  • the drug may be coated with a time-disintegrating material, or may be incorporated into an appropriate drug release system. It may be formulated with a targeting agent for delivery to the target cancer tissue.
  • the cancer therapeutic agent of the present invention can be administered by various routes including both oral and parenteral, such as, but not limited to, oral, intravenous, intramuscular, subcutaneous, topical (particularly, intratumoral), lymphatic or Intralymph node, intrathecal, rectal, intraarterial, intraportal, intraventricular, transmucosal, transdermal, intranasal, intraperitoneal, intrapulmonary, intrauterine, etc. It can be formulated into a dosage form suitable for the route. As such a dosage form and a preparation method, any known one can be appropriately adopted (see, for example, Standard Pharmacology, edited by Yoshiteru Watanabe et al., Nankodo, 2003).
  • an effective amount of an inhibitor of fibrillarin expression or activity is, for example, an amount that suppresses cancer growth, reduces symptoms, or delays or stops progression, and preferably inhibits cancer progression. Or the amount that cures the cancer. Also preferred is an amount that does not cause adverse effects beyond the benefit of administration.
  • Such an amount can be appropriately determined by an in vitro test using cultured cells or the like, or a test in a model animal such as mouse, rat, dog or pig, and such a test method is well known to those skilled in the art. . Further, the dose of the additive or the carrier is also known to those skilled in the art, or can be appropriately determined by the above-described test and the like.
  • the present invention provides a method for suppressing cancer cell proliferation, which comprises administering to a cancer patient an inhibitor of the expression or activity of fibrillarin in an amount effective for suppressing cancer cell growth.
  • Suppressing the growth of cancer cells refers to reducing the degree of proliferation of cancer cells as compared to the case where no drug is administered.
  • the present invention also provides a method for treating cancer, which comprises administering to a cancer patient an inhibitor of the expression or activity of fibrillarin in an amount effective for treating cancer.
  • Cancer treatment includes cancer cure, remission, amelioration, mitigation, progression delay, progression arrest, and the like.
  • the “cancer cell” of the present invention refers to a cancer cell with high fibrillarin expression.
  • the “cancer patient” of the present invention is a patient with a high fibrillarin-expressing cancer cell, that is, a prognosis. You have a bad cancer patient.
  • the inhibitor of fibrillarin expression or activity may be formulated and administered together with additives and carriers.
  • the route of administration differs depending on the dosage form, and is oral, intravenous, intramuscular, subcutaneous, local (particularly, within a tumor), lymphatic vessel or lymph node, intrathecal, rectum, intraarterial, intraportal, intraventricular. Transmucosal, transdermal, intranasal, intraperitoneal, intrapulmonary, intrauterine, etc. routes.
  • the amount of the inhibitor of the expression or activity of fibrillarin, which is effective for suppressing the growth of cancer cells or treating cancer depends on various conditions, for example, the type of cancer, the severity of symptoms, general health, age, and body weight. The determination can be made by those skilled in the art in consideration of, for example, gender, diet, timing and frequency of administration, concomitant medication, response to treatment, compliance with treatment, dosage form, and administration route.
  • the amount of the inhibitor of the expression or activity of fibrillarin, which is effective for suppressing the growth of cancer cells or for treating cancer varies depending on the above-mentioned various conditions, but is, for example, 0.00001 mg to 100 g per day, preferably 0.0001 mg per day.
  • the dose can be from 10 to 10 g, especially from 0.001 mg to 1 g, from 0.01 mg to 100 mg, especially from 0.1 mg to 10 mg.
  • the dose is an amount calculated as a nucleic acid compound.
  • the daily dose is calculated from the total dose.
  • the dosing frequency or schedule will depend on the various conditions described above, but may be, for example, many times a day (ie, 2, 3, 4, or 5 or more times a day), once a day, every few days (ie, 2, 3 Every four, five, six, seven days, etc.) several times a week (for example, two, three, four times a week, etc.) every one week, every few weeks (ie, every two, three, four weeks, etc.) It may be. Further, the administration frequency may change over time depending on the symptoms. The period of administration can be until the cancer is cured or ameliorated.
  • the present invention provides a step of measuring the fibrillarin expression level of a cancer tissue of a cancer patient, a step of comparing the measured value with a median fibrillarin expression level predetermined for each type of cancer, and the measured value. Determining that the prognosis is poor when the measured value is higher than or equal to the median value, and determining that the prognosis is good when the measured value is lower than the median value.
  • the fibrillarin expression level ie, the fibrillarin gene expression level, can be determined by extracting RNA from a cancer tissue collected from a cancer patient in accordance with a conventional method, for example, by RT-qPCR.
  • the median fibrillarin expression level of various cancer cells refers to the median fibrillarin expression level in cancer tissues of at least 50, especially 50 to 150, cancer patients. It may be determined from data collected in the Cancer Genome Atlas Project by the Human Genome Research Institute.
  • Poor prognosis means that the survival rate is low after the start of cancer treatment (including surgical treatment, including surgery such as cancer resection, radiation therapy, and chemotherapy). It means that the survival rate after the day of starting such cancer treatment is high.
  • the survival rate can be, for example, the survival rate on the 50th to 3000th day, especially the 300th to 1500th day from the day when the cancer treatment is started.
  • 50 to 150 days (especially 50 days) from the start of cancer treatment for colorectal cancer, 50 to 150 days (especially 50 days) from the start of cancer treatment for ovarian cancer, and cancer treatment for breast cancer 50 to 150 days (especially 100 days) from the day, 1000 to 3000 days (especially 1500 days) from the start of cancer treatment for lung adenocarcinoma, and 300 to 1000 days from the start of cancer treatment for glioma.
  • the survival rate after days can be used.
  • the fibrillarin expression level may be measured either before or after the cancer treatment start date. Above all, it is preferable to measure the fibrillarin expression level before the start of the cancer treatment. In the case of the cancer treatment by suppressing the fibrillarin expression, the fibrillarin expression level may be measured before the start of the cancer treatment.
  • Fibrillarin gene expression level in various cancer-bearing patients a) Materials and reagents: Public data such as cancer genomes, epigenomes, transcriptomes, and mutation information in various tissues, collected from the National Cancer Institute's Cancer Genome Atlas Project by the US National Cancer Institute b) Methods: RNA expression data (RPKM value: Reads per kilobase of exon per million mapped reads) of the fibrillarin gene analyzed by RNA-seq in the public data was obtained, and the relative value of the fibrillarin RNA expression level was obtained using the expression level of all genes. (AU) was calculated by the normalization method, and the relative value of the fibrillarin gene expression level for each patient was displayed as log2. c) Result: As shown in FIG. 1, it was revealed that the fibrillarin gene was highly expressed in many cancers.
  • Fibrillarin expression level (production level) inhibitory effect of fibrillarin siRNA in renal cell carcinoma cell line a) Materials and reagents: Renal cell carcinoma: 7860 strain fibrillarin siRNA: The nucleotide sequence of the RNA corresponding to positions 103 to 121 in the nucleotide sequence of the human fibrillarin gene (SEQ ID NO: 1: ggucgaggcgggagcucuua) is used.
  • SEQ ID NO: 1: ggucgaggcgggagcucuua ggucgaggcgggagcucuua
  • Method After culturing the 7860 strain on a 24-well plate, 5 ⁇ M of fibrillarin siRNA was introduced into the cells using 5 ⁇ L of Dharmafect reagent (GE-healthcare).
  • the fibrillarin expression level was expressed as a relative value to the fibrillarin expression level of control cells into which siRNA was not introduced.
  • c) Result As shown in FIG. 7, the use of fibrillarin siRNA reduced the fibrillarin expression level (production level) of the renal cell carcinoma cell line to about 1/10.
  • Example 4 Test for inhibiting the growth of xenograft renal cell carcinoma by suppressing the fibrillarin expression level (production level)
  • Materials and reagents -Human kidney cancer cell line for xenotransplantation: 786 Mock -Fibrillarin siRNA: Uses the nucleotide sequence of the human fibrillarin gene (the nucleotide sequence corresponding to the nucleotide sequence at positions 103 to 121 (SEQ ID NO: 1: ggucggggcgggagcucuua)).
  • -EGFP siRNA MISSION siRNA (SIGMA) is used.
  • kidney cancer formation was suppressed at the site where fibrillarin siRNA was administered.
  • the diameter of transplanted kidney cancer was clearly reduced, and it is considered that the cancer had died.
  • the diameter of transplanted kidney cancer was increased.
  • the administration and administration of fibrillarin siRNA significantly suppressed the engraftment and proliferation of cancer cells in nude mice by suppressing the expression of fibrillarin.
  • the suppression of fibrillarin expression or the suppression of fibrillarin action in cancer cells, particularly cancer cells highly expressing fibrillarin can suppress the growth of cancer cells. That is, it indicates that cancer treatment is possible by suppressing the expression of fibrillarin or suppressing the action of fibrillarin.
  • the inhibitor of the expression or action of fibrillarin of the present invention effectively suppresses the growth of cancer cells in cancer cells, particularly cancer cells in which fibrillarin is highly expressed, and provides a new mechanism of action for cancer treatment. You can do it.
  • the inhibitor of the expression or action of fibrillarin of the present invention can be used as a single agent, and can also be used in combination with other cancer therapeutic agents.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Molecular Biology (AREA)
  • Reproductive Health (AREA)
  • Biomedical Technology (AREA)
  • Pulmonology (AREA)
  • Endocrinology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Biochemistry (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Immunology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Le présent inventeur a découvert que, dans une maladie cancéreuse, les cellules cancéreuses présentant une grande quantité d'expression (quantité de production) de fibrillarine sont associées à un mauvais pronostic (un faible taux de survie). En outre, le présent inventeur a découvert que la prolifération de cellules cancéreuses peut être inhibée par la réduction de la quantité d'expression (quantité de production) de fibrillarine. Par conséquent, il devient possible d'inhiber la prolifération de cellules cancéreuses à mauvais pronostic pour ainsi traiter une maladie cancéreuse à l'aide de l'inhibiteur de l'expression ou de l'activité de fibrillarine selon la présente invention. Des exemples caractéristiques de l'inhibiteur de l'expression ou de l'activité de fibrillarine comprennent un ARNsi, un ARN court en épingle à cheveux, un antisens, etc.
PCT/JP2019/038825 2018-10-02 2019-10-01 Inhibiteur de prolifération de cellules cancéreuses à mauvais pronostic WO2020071392A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2020550471A JP7191301B2 (ja) 2018-10-02 2019-10-01 予後不良のがん細胞の増殖抑制剤

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2018198764 2018-10-02
JP2018-198764 2018-10-02

Publications (1)

Publication Number Publication Date
WO2020071392A1 true WO2020071392A1 (fr) 2020-04-09

Family

ID=70054813

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2019/038825 WO2020071392A1 (fr) 2018-10-02 2019-10-01 Inhibiteur de prolifération de cellules cancéreuses à mauvais pronostic

Country Status (2)

Country Link
JP (1) JP7191301B2 (fr)
WO (1) WO2020071392A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006507841A (ja) * 2002-11-14 2006-03-09 ダーマコン， インコーポレイテッド 機能的ｓｉＲＮＡおよび超機能的ｓｉＲＮＡ

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006507841A (ja) * 2002-11-14 2006-03-09 ダーマコン， インコーポレイテッド 機能的ｓｉＲＮＡおよび超機能的ｓｉＲＮＡ

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
KOH, C.M. ET AL.: "Alterations in Nucleolar Structure and Gene Expression Programs in Prostatic Neoplasia Are Driven by the MYC Oncogene", THE AMERICAN JOURNAL OF PATHOLOGY, vol. 178, no. 4, 2011, pages 1824 - 1834, XP055699395, ISSN: 0002-9440 *
MARCEL, V. ET AL.: "Acts as a Safeguard of Translational Control by Regulating Fibrillan and rRNA Methylation in Cance", CANCER CELL, vol. 24, 2013, pages 318 - 330, XP028712570, ISSN: 1535-6108, DOI: 10.1016/j.ccr.2013.08.013 *
SU , H. ET AL.: "Elevated snoRNA biogenesis is essential in breast cancer", ONCOGENE, vol. 33, 2014, pages 1348 - 1358, XP055699394, ISSN: 0950-9232 *

Also Published As

Publication number Publication date
JP7191301B2 (ja) 2022-12-19
JPWO2020071392A1 (ja) 2021-09-02

Similar Documents

Publication Publication Date Title
US20200056177A1 (en) Long non-coding rna used for anticancer therapy
JP2021020951A (ja) セロトニン放出ホルモン、アドレナリン放出ホルモン、ノルアドレナリン放出ホルモン、グルタミン酸放出ホルモンおよび副腎皮質刺激ホルモン放出ホルモン関連の医学的状態の処置および診断のためのマイクロｒｎａおよび該マイクロｒｎａを含む組成物
US10894989B2 (en) Treatment of angiogenesis disorders
KR20110026023A (ko) 아이지에프비피-2 및 아이지에프비피-5를 억제하는 양특이성 안티센스 올리고뉴클레오티드 및 그 사용방법
JP2020534354A (ja) トリプルネガティブ乳癌の治療方法
IL277136A (en) Means and methods for lowering the tumorigenicity of cancer stem cells
WO2017156591A1 (fr) Miarn d'ostéo-arthrite
JP2009541304A5 (fr)
JP2009541304A (ja) 抗癌治療としてのオーロラキナーゼｂのｒｎａ干渉媒介阻害およびその組み合わせ
WO2020071392A1 (fr) Inhibiteur de prolifération de cellules cancéreuses à mauvais pronostic
KR101197627B1 (ko) Ｈｐｖ 감염과 관련된 암의 치료용 조성물
JP5180084B2 (ja) がん細胞識別マーカー及びがん細胞増殖阻害剤
JPWO2015156237A1 (ja) 慢性腎臓病治療用医薬組成物
JP2021028297A (ja) マイクロｒｎａ及びその誘導体を有効成分とするがん治療剤
Liu et al. Circ_0021573 acts as a competing endogenous RNA to promote the malignant phenotypes of human ovarian cancer cells
WO2021132309A1 (fr) Composition pharmaceutique
JP7137183B2 (ja) がんの治療又は予防用医薬および癌のバイオマーカー
WO2020040185A1 (fr) Médicament mettant en œuvre un inhibiteur de hsp47 pour augmenter la sensibilité à un médicament chimiothérapeutique
WO2017159739A1 (fr) Agent anticancéreux
Hubbard Transcription factors as novel targets for cancer therapy
WO2014024820A1 (fr) Composition pharmaceutique destinée au traitement d'une nouvelle maladie rénale chronique et procédé de criblage de médicaments destinés à la nouvelle maladie rénale chronique

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19869024

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2020550471

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19869024

Country of ref document: EP

Kind code of ref document: A1