WO2014129590A1 - INHIBITEUR DE L'EXPRESSION DU GÈNE DE LA Cycline D1 ET MÉDICAMENT ANTICANCÉREUX ASSOCIÉ - Google Patents

INHIBITEUR DE L'EXPRESSION DU GÈNE DE LA Cycline D1 ET MÉDICAMENT ANTICANCÉREUX ASSOCIÉ Download PDF

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WO2014129590A1
WO2014129590A1 PCT/JP2014/054196 JP2014054196W WO2014129590A1 WO 2014129590 A1 WO2014129590 A1 WO 2014129590A1 JP 2014054196 W JP2014054196 W JP 2014054196W WO 2014129590 A1 WO2014129590 A1 WO 2014129590A1
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peptide
amino acid
cells
seq
cyclin
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PCT/JP2014/054196
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Japanese (ja)
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杉山　大介
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国立大学法人九州大学
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    • 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
    • A61K38/10—Peptides having 12 to 20 amino acids
    • 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 Cyclin D1 gene expression inhibitor and an antitumor agent comprising a peptide having a specific amino acid sequence.
  • the number of cancer patients in the world is approximately 12.7 million / year, and the number of cancer patients in Japan is approximately 670,000 / year.
  • the leading cause of death among Japanese is “cancer” (approximately 340,000 people / year).
  • cancer multidisciplinary treatment using surgical operation, chemotherapy, radiation therapy, etc. is performed, but the current situation is that cancer has not yet been suppressed.
  • anticancer agents are organic compounds having a molecular weight of 200 to 400 that directly act on nucleic acid synthesis and have a cell-killing effect. Therefore, when such an anticancer agent is administered, normal cells, particularly normal cells that are actively proliferating, are affected, and it is difficult to avoid strong side effects.
  • Patent Document 1 proposes an Akt activity-inhibiting polypeptide.
  • Patent Document 2 discloses a therapeutic cancer peptide vaccine. Since the mechanism of action of such peptides with anticancer activity is similar to that of molecular targeted drugs, it is expected as a new anticancer agent. It is rare.
  • An object of the present invention is to provide a Cyclin D1 gene expression inhibitor and an antitumor agent capable of slowly suppressing the proliferation of cancer cells without having a cytocidal action and preventing recurrence of cancer as a new anticancer agent. That is.
  • the present inventor has previously promoted research focusing on embryonic liver hematopoiesis, which is an organ in which hematopoietic stem cells actively proliferate and differentiate, and as a result, the amino acid sequence of SEQ ID NO: 1 suppresses proliferation and differentiation of hematopoietic stem cells.
  • a novel physiologically active peptide having a molecular weight (molecular weight: 1348.5; number of amino acids: 13) (International Publication No. 2011/040500).
  • KS-13 is taken up by endocytosis in the added culture and suppresses the proliferation and differentiation of mouse / human hematopoietic cells (hereinafter, the peptide is also referred to as “KS-13”).
  • KS-13 and its derivatives can be added to cells of various cancer cell lines such as leukemia cell lines, prostate cancer cell lines, breast cancer cell lines. It was found that by suppressing the expression of the Cyclin D1 gene, which controls the cycle, to 1/100 or less, the growth of cell lines was suppressed by about 20%, that is, KS-13 and its derivatives had anticancer activity. The invention has been completed.
  • the Cyclin D1 gene expression inhibitor of the present invention is an active ingredient, (A) a peptide having the amino acid sequence shown in SEQ ID NO: 1; or (B) a peptide obtained by deleting, substituting, or adding at least one amino acid residue in the amino acid sequence shown in SEQ ID NO: 1, A cyclin D1 gene expression inhibitor comprising a peptide having a D1 gene expression inhibitory action.
  • a preferred embodiment of the present invention is the above Cyclin D1 gene expression inhibitor
  • the peptide of (B) is a Cyclin D1 gene expression inhibitor, which is a peptide having the amino acid sequence shown in SEQ ID NO: 2.
  • a preferred embodiment of the present invention is the above Cyclin D1 gene expression inhibitor
  • the peptide of (B) is a Cyclin D1 gene expression inhibitor, which is a peptide having any one amino acid sequence selected from SEQ ID NO: 3 to SEQ ID NO: 10.
  • the antitumor agent of the present invention is an active ingredient, (A) a peptide having the amino acid sequence shown in SEQ ID NO: 1; or (B) a peptide obtained by deleting, substituting, or adding at least one amino acid residue in the amino acid sequence shown in SEQ ID NO: 1, An antitumor agent comprising a peptide having a D1 gene expression inhibitory action.
  • the peptide of (B) is an antitumor agent which is a peptide having the amino acid sequence shown in SEQ ID NO: 2.
  • the peptide (B) is an antitumor agent which is a peptide having any one amino acid sequence selected from SEQ ID NO: 3 to SEQ ID NO: 10.
  • a peptide having the amino acid sequence shown in SEQ ID NO: 1 or (B) at least one amino acid residue in the amino acid sequence shown in SEQ ID NO: 1 is deleted or substituted
  • a peptide that is added and has a cyclin D1 gene expression inhibitory action it slowly suppresses the growth of cancer cells without having a cytocidal action, and also prevents recurrence of cancer as a new anticancer agent. Cyclin D1 gene expression inhibitor and antitumor agent that can be prevented can be provided.
  • FIG. 1 is a graph showing the results of Experimental Example 1.
  • FIG. 2 is a photomicrograph showing the results of Experimental Example 2-1.
  • FIG. 3 is a photomicrograph showing the results of Experimental Example 2-2.
  • FIG. 4 is a graph showing the results of Experimental Example 3-1.
  • FIG. 5 is a graph showing the results of Experimental Example 3-2.
  • FIG. 6 is a graph showing the results of Experimental Example 3-3.
  • FIG. 7 is a graph showing the results of Experimental Example 4 (Example 1).
  • FIG. 8 is a photomicrograph showing the results of Experimental Example 5-1.
  • FIG. 9 is a diagram showing the results of Experimental Example 5-2.
  • FIG. 10 is a diagram illustrating the results of Experimental Example 6.
  • FIG. 11 is a diagram showing the results of Experimental Example 7-1.
  • FIG. 12 is a diagram showing the method and results of Experimental Example 7-2.
  • FIG. 13 is a diagram showing the method and results of Experimental Example 7-2.
  • FIG. 14 is a graph showing the results of Experimental Example 8-1.
  • FIG. 15 is a graph showing the results of Experimental Example 8-2.
  • FIG. 16 is a graph showing the results of Experimental Example 9-1.
  • FIG. 17 is a graph showing the results of Experimental Example 9-2.
  • FIG. 18 is a graph showing the results of Experimental Example 9-3.
  • FIG. 19 is a graph showing the results of Experimental Example 9-4.
  • FIG. 20 is a graph showing the results of Experimental Example 9-5.
  • FIG. 21 is a graph showing the results of Experimental Example 9-6.
  • FIG. 22 is a graph showing the results of Experimental Example 10 (Example 2).
  • Cyclin D1 gene is a gene encoding a cell cycle regulator and is also known as a so-called “cancer-related gene”. When expressed in normal cells, the gene is capable of tumorigenic or cancerous, and Cyclin D1 gene overexpression is caused by parathyroid adenoma, breast cancer, prostate cancer, colon cancer, lymphoma, melanoma And involved in the development of lung cancer (Morgan, D.O., (2008), Cell, 135,764-794, Lung Cancer, (2007), 55, 1-14).
  • a “gene expression inhibitor” is a drug that generally suppresses the expression of a target gene at the mRNA level or protein level. Although the mechanism of action of Cyclin D1 gene expression suppression in the gene expression inhibitor of the present invention is still unclear, it cannot be suppressed by binding a peptide having Cyclin D1 gene expression inhibitory action directly to the promoter region.
  • S12A3 Solute carrier family12 member3
  • Antineoplastic agent is a chemotherapeutic agent that is generally administered for the purpose of treating tumors in a broad sense.
  • the antitumor agent of the present invention slowly suppresses the growth of cancer cells by suppressing the expression of the cancer-related gene Cyclin D1 gene that causes normal cells to become tumorous or cancerous, and is not due to cell killing action. Therefore, it has a feature that strong side effects can be avoided.
  • the peptide targeted by the present invention is a peptide having a Cyclin D1 gene expression inhibitory action.
  • One embodiment of such a peptide is a peptide having an amino acid sequence consisting of the amino acid residues shown below:
  • the peptide consists of a partial sequence of the extracellular domain located at positions 24 to 303 of human Dlk1 (delta-like 1 homolog) protein (SEQ ID NO: 11) consisting of 383 amino acid residues in total length. As mentioned above, this peptide is also referred to herein as “KS-13”.
  • the peptide targeted by the present invention is a peptide obtained by deleting, substituting or adding at least 1, preferably about 1 to 6 amino acids in the amino acid sequence of KS-13 (SEQ ID NO: 1). And a peptide having an inhibitory effect on the expression of Cyclin D1 gene.
  • the peptide targeted by the present invention is as described above, but it is 60% or more, preferably 75% or more, more preferably 80% or more, and still more preferably 85% to 90% or more. It preferably has the same or similar amino acid sequence as shown in.
  • the ratio of “identity” or “similarity” can be calculated from the ratio of identical or similar amino acids overlapping with the total number of amino acid residues of the amino acid sequence shown in SEQ ID NO: 1.
  • similar amino acids mean amino acids similar in physicochemical properties.
  • similar amino acids include aromatic amino acid groups (Phe, Trp, Tyr), aliphatic amino acid groups (Ala, Leu, Ile).
  • Examples of the peptide obtained by substitution, deletion or addition include a peptide represented by the following amino acid sequence (SEQ ID NO: 2) and having the above action.
  • peptides corresponding to KS-13 described above consisting of a partial sequence of the extracellular domain region of a protein corresponding to an ortholog of human Dlk1 protein.
  • mouse Dlk1 protein corresponding to an ortholog of human Dlk1 protein has an amino acid sequence consisting of a total length of 385 amino acid residues (SEQ ID NO: 12).
  • SEQ ID NO: 12 amino acid sequence consisting of a total length of 385 amino acid residues
  • the region at positions 24 to 305 is the extracellular domain
  • the partial sequence sequence consisting of 13 amino acid residues at positions 124 to 136: SEQ ID NO: 3
  • the partial sequence corresponds to KS-13. It is a peptide.
  • mice In addition to mice, rodents such as rats and rats, and mammals such as pigs, horses, goats, cattle, wallabies, and giant kangaroos; It has Dlk1 protein corresponding to protein homologue or orthologue.
  • the partial sequences corresponding to the above KS-13 located in the extracellular domain of these Dlk1 proteins are shown in Table 1 together with the amino acid sequences of peptides derived from human-derived KS-13 and mouse KS-13. Amino acid residues conserved in common with human-derived KS-13 are indicated by underlining.
  • the third amino acid from the N-terminus (corresponding to “XXb” in the amino acid sequence of SEQ ID NO: 2) may be either Lys or His. It can be seen that the fifth amino acid (corresponding to “XXd” in the amino acid sequence of SEQ ID NO: 2) may be either Asp or Ala. Further, comparing the sequences of human and porcine peptides, it can be seen that the 10th amino acid from the N-terminal (corresponding to “XXf” in the amino acid sequence of SEQ ID NO: 2) may be either Ile or Met.
  • the third amino acid from the N-terminus may be deleted, and the fifth amino acid from the N-terminus (“XXd” in the amino acid sequence of SEQ ID NO: 2). It is understood that either Asp or Glu may be used.
  • the 5th amino acid from the N-terminus may be deleted, and the 9th amino acid from the N-terminus (in the amino acid sequence of SEQ ID NO: 2, “XXe”). It can be seen that any of Val and Ile can be used.
  • KS-13 is the most preferred peptide, but as shown in SEQ ID NO: 2, any one of amino acids 1 to 6 (XXa to XXf) in the amino acid sequence is another amino acid. It may be substituted, may be a peptide consisting of 12 amino acid residues with one less N-terminal or C-terminal amino acid, or a peptide consisting of 11 amino acid residues with one less amino acid Good.
  • the N-terminal amino acid may be deleted up to 5 or 4 to 3 amino acids.
  • the peptide formed by adding at least one amino acid has five amino acids derived from human Dlk1 protein at the N-terminus and / or C-terminus of the amino acid sequence, Preferred are those with 4, 3, 2 or 1 added.
  • a method for substituting, adding or deleting at least one amino acid in the amino acid sequence of KS-13 has already been conventionally used in the art.
  • DNA encoding the peptide In the case of performing via the Internet, for example, site-specific mutagenesis [Methods in Enzymology, 154, 350, 367-382 (1987); 100, 468 (1983); Nucleic Acids Res. , 12, 9441 (1984); secondary biochemistry experiment course 1 "Gene Research Method II", edited by Japanese Biochemical Society, p105 (1986)], etc., phosphate triester method, phosphate amidite method, etc. Chemical synthesis means [J. Am. Chem. Soc.
  • DNA synthesis can be performed by chemical synthesis by the phosphoramidite method or triester method, and can also be performed on a commercially available automatic oligonucleotide synthesizer.
  • Double-stranded fragments are chemically synthesized single strands produced by synthesizing complementary strands and annealing the strands together under appropriate conditions, or adding complementary strands using DNA polymerase with appropriate primer sequences. It can also be obtained from things.
  • the peptide according to the present invention can also be synthesized by a solid phase synthesis method using a peptide synthesizer, and amino acid substitution, addition or deletion changes the type of protected amino acid when using a peptide synthesizer. This can be done easily.
  • special amino acids such as D-amino acid and sarcosine (N-methylglycine) can be introduced.
  • the peptide according to the present invention may be in a free state, in the form of a salt, or in the form of a solvate containing a hydrate.
  • the salt include physiologically acceptable, that is, pharmaceutically acceptable acid addition salts and base salts.
  • acid addition salts include inorganic acid salts such as hydrochloride, hydrobromide, nitrate and sulfate; or sulfonic acid salts such as methanesulfonic acid and toluenesulfonic acid; organic acids such as trifluoroacetic acid and succinic acid.
  • Examples of the base salt include alkali metal salts such as sodium, potassium and lithium; or alkaline earth metal salts such as calcium and magnesium.
  • the peptides according to the present invention include peptides whose C-terminus is a carboxyl group (—COOH), carboxylate (—COO—), amide (—CONH 2 ), or ester (—COOR).
  • R in the ester for example, an alkyl group having 1 to 6 carbon atoms such as methyl, ethyl, n-propyl, isopropyl and n-butyl; a cycloalkyl group having 3 to 8 carbon atoms such as cyclopentyl and cyclohexyl; phenyl, aryl groups having 6 to 12 carbon atoms such as ⁇ -naphthyl; phenyl-C1-2 alkyl groups such as benzyl and phenethyl; C7-14 aralkyl groups such as ⁇ -naphthyl-C1-2 alkyl groups such as ⁇ -naphthylmethyl; A pivaloyloxymethyl group
  • the peptide according to the present invention when the peptide according to the present invention has a carboxyl group (or carboxylate) other than the C-terminus, the peptide according to the present invention includes those in which the carboxyl group is amidated or esterified.
  • the amino group of the N-terminal amino acid residue is protected with a protecting group (for example, a C1-6 acyl group such as C1-6 alkanoyl such as formyl group and acetyl group).
  • fatty acid saturated fatty acid of C8-18
  • N-terminal glutamine residue which can be cleaved in vivo, pyroglutamine oxidized, substituent on side chain of amino acid in molecule
  • suitable protecting groups for example, C1-6 acyl groups such as C1-6 alkanoyl groups such as formyl group, acetyl group, etc.
  • a complex peptide such as a so-called glycopeptide to which a sugar chain is bound.
  • the peptides according to the present invention include those in which an imidazolyl group or SH group is alkylated (for example, methylated), aralkylated (for example, benzylated), or acylated (for example, acetylated or benzoylated).
  • the fatty acid-modified products include myristoylated peptides in which the amino group of the N-terminal amino acid residue is modified with myristic acid.
  • Cyclin D1 gene expression inhibitor The peptide according to the present invention detailed above or a pharmaceutically acceptable salt or solvate thereof as it is or with a carrier that is physiologically acceptable as necessary After mixing to make a composition, it is used as the Cyclin D1 gene expression inhibitor of the present invention.
  • the peptide based on this invention consists of 1 type individually, 2 or more types can also be used in arbitrary combinations.
  • human-derived KS-13 (SEQ ID NO: 1), rodent-derived peptide (SEQ ID NO: 3), porcine-derived peptide (SEQ ID NO: 4), chicken-derived peptide (SEQ ID NO: 10), and more Preferred are human-derived KS-13 (SEQ ID NO: 1) and rodent-derived peptides (SEQ ID NO: 3).
  • Cyclin D1 gene expression inhibitor dissolves the peptide of the present invention in water or an appropriate buffer solution (eg, phosphate buffer solution, PBS, Tris-HCl buffer solution, etc.) to an appropriate concentration. Can be prepared. Moreover, you may mix
  • an appropriate buffer solution eg, phosphate buffer solution, PBS, Tris-HCl buffer solution, etc.
  • (IV) Antitumor Agent The peptide according to the present invention detailed above or a pharmaceutically acceptable salt or solvate thereof is mixed as it is or with a physiologically acceptable carrier as necessary. After preparing the composition, it is used as the antitumor agent of the present invention. In addition, even if the peptide based on this invention consists of 1 type individually, 2 or more types can also be used in arbitrary combinations.
  • human-derived KS-13 (SEQ ID NO: 1), rodent-derived peptide (SEQ ID NO: 3), porcine-derived peptide (SEQ ID NO: 4), chicken-derived peptide (SEQ ID NO: 10), and more Preferred are human-derived KS-13 (SEQ ID NO: 1) and rodent-derived peptides (SEQ ID NO: 3).
  • the antitumor agent is prepared, for example, by dissolving the peptide according to the present invention in water or an appropriate buffer (eg, phosphate buffer, PBS, Tris-HCl buffer, etc.) to an appropriate concentration. can do. Moreover, you may mix
  • an appropriate buffer eg, phosphate buffer, PBS, Tris-HCl buffer, etc.
  • PC-3 cells are adherent cells, the cells are collected by adding 0.2% trypsin and then with RPMI 1640 + 10% fetal bovine serum (FBS) supplemented with 10 ⁇ g / ml KS-13.
  • FBS fetal bovine serum
  • a cell suspension of ⁇ 10 5 cells / ml was cultured in a 48-well tissue culture plate. Cultures without KS-13 were used as controls. One day, two days, and five days later, cells stained with trypan blue and unstained live cells were counted under a microscope. After 5 days, a photograph of the cell line morphology was taken under a microscope.
  • KS-13 was added to the culture medium of the K562 cell line and monitored with a horoscope microscope for 30 minutes. The results are shown in FIG. White dots indicate the KS-13 added group, and black dots indicate the control (KS-13 non-added group). That is, when KS-13 was added, differentiation of the K562 cell line was induced, the thickness changed, and it became clear that it became an irregular form. Thus, it is considered that cancer cell proliferation is suppressed by KS-13.
  • FIG. 5 shows the results of representing irregularity on the vertical axis and roughness on the horizontal axis.
  • FIG. 6 shows the results of representing irregularity on the vertical axis and roughness on the horizontal axis.
  • RNAqueous-4PCR (trademark) (manufactured by Life Technologies) according to the product protocol.
  • the mRNA was reverse transcribed using High Capacity RNA-to-cDNA Kit (manufactured by Life Technologies).
  • the quality of cDNA synthesis was checked by PCR amplification of ⁇ -actin (ACTB), a human housekeeping gene.
  • ACTB ⁇ -actin
  • KS-13 increased the expression of Cyclin D1 (CCND1) gene in human leukemia cell line (Kasumi), human prostate cancer cell line (PC-3), and human breast cancer cell line (T47D) by 141 times, It was found to be suppressed 122 times and 152 times.
  • CCND1 Cyclin D1
  • Chromatin immunoprecipitation A 2 ⁇ 10 5 cell / ml K562 cell suspension was cultured for 2 days using RPMI 1640 + 10% fetal bovine serum (FBS) supplemented with 10 ⁇ g / ml biotin-conjugated KS-13.
  • the number of cells used for ChIP is 1 ⁇ 10 6 cells.
  • FBS fetal bovine serum
  • ChIP ChIP lysis buffer containing SDS. Cell debris was removed by centrifugation at 12000 rpm for 10 minutes (4 ° C.). Chromosomal DNA was fragmented by sonication (250 W, 5 min / 30 sec on / off cycle) to obtain chromosomal DNA of a small size of 100-1000 bp. ChIP was performed according to the product instructions (Chromatin immunoprecipitation assay kit, MILLIPORE). That is, the fragmented DNA was diluted with a ChIP binding buffer. Genomic DNA diluted to 1% was used as a control “before ChIP”. Rabbit anti-biotin antibody was used to precipitate KS-13 and genomic DNA bound to KS-13. The precipitated chromosomal DNA was used as a “post-ChIP” template for PCR.
  • PCR A 127 bp PCR product was obtained using a primer that binds to the promoter region of human Cyclin D1.
  • K562 genomic DNA was used as a positive control for PCR, and no template was used as a negative control.
  • the PCR product was electrophoresed on a 2% agarose gel simultaneously with a DNA ladder marker for 100 bp and stained with SYBER Green.
  • FBS fetal bovine serum
  • the number of cells used for ChIP is 1 ⁇ 10 6 cells. After washing with cold PBS, the cells were lysed using an IP lysis buffer (Pierce Classic IP Kit, Thermo Scientific).
  • IP was performed according to the product instructions (Pierce Classic IP Kit, Thermo Scientific). Rabbit anti-biotin antibody was used to precipitate KS-13 and KS-13 binding proteins. The biotin-KS-13 protein complex was separated from unbound protein by adding protein G. After three washes, the protein was separated from the immune complex using low PH buffer. The eluted protein comprises heavy chain (50 kDa) and light chain (25 kDa) rabbit IgG. The eluted protein was electrophoresed on SDS-PAGE and stained with Coomassie Brilliant blue G-250 (Bio-Rad).
  • Example 7-1 Identification of K-13 binding protein in K562 cells was performed using liquid chromatography mass spectrometry (LC-MS). As a result, ATP-dependent RNA helicase (YTHDC2) and single carrier family12 member3 (S12A3) seemed to bind to K-13 in K562 cells compared to the control without addition of K-13. (See FIG. 11). Thus, two target molecules of KS-13 in the K562 leukemia cell line were revealed.
  • LC-MS liquid chromatography mass spectrometry
  • LC-MS Liquid chromatograph mass spectrometry
  • Identified K-13 binding protein Dermcidin (DCD) and U2 small nuclear ribonucleoproteins A and B (RU2A, RU2B) were identified as K-13 binding proteins of K562 cells.
  • High mobility group protein 2 (HMGN2) and SUB1 homologue (S. cerevisiae) (SUB1) binding to nucleosomes were identified as K-13 binding proteins of Kasumi cells.
  • Example 8-2 (1) Materials and Methods A suspension of 2 ⁇ 10 5 cells / ml of K562 cells was cultured for 2 days using RPMI + 10% fetal bovine serum (FBS) supplemented with 10 ⁇ g / ml of KS-13. A phosphorylated proteome array (phosphorylation-kinase array kit, R & D Systems) was performed according to the product instructions. (2) Results There was no difference in protein phosphorylation of K562 cells treated with KS-13 (see FIG. 15).
  • FBS fetal bovine serum
  • KS-13 Low low concentration 0.36 ⁇ g / g body weight
  • KS-13 High high concentration 3.6 ⁇ g / g body weight
  • leukocytes The number of WBC
  • RBC red blood cells
  • PHT platelets
  • HGB hemoglobin
  • GOT Glutamate oxaloacetate transaminase
  • GPT Glutamate pyruvate transaminase
  • KS-13 (a peptide having the amino acid sequence shown in SEQ ID NO: 1) suppresses the growth of leukemia cell line / prostate cancer cell line, leukemia cell line / breast cancer cell line / prostate It was confirmed that the CyclinD1 gene expression of the cell line was suppressed, that it did not act directly on the CyclinD1 gene expression region, that there were two molecular targets in the K562 leukemia cell line, and that no toxicity was observed after a single administration.
  • Non-obese diabetic mice are transferred intravenously with 2 ⁇ 10 7 human myeloid leukemia cell line (Kasumi).
  • PBS fetal bovine serum
  • 5FU anticancer agent
  • 5FU + 72 ⁇ g KS-13 were intravenously administered.
  • the survival of the mice was observed every week. The survival rate for each week was plotted on the graph (see FIG. 22). Dead mice were fixed with 4% formaldehyde for further analysis.
  • mice administered with PBS died.
  • mice administered KS-13 were 100% alive until the 41st week.
  • a tumor-bearing mouse using the Kasumi leukemia cell line was prepared and the anticancer effect of a single administration of KS-13 was evaluated, a clear life-prolonging effect was observed.
  • the present invention has high technical significance as 1) can contribute to the survival of cancer patients as a new drug; 2) can contribute to target molecule identification of a new drug; 3) can contribute to life science as a research reagent.

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Abstract

L'invention concerne un inhibiteur de l'expression du gène de la cycline D1, ainsi qu'un médicament anticancéreux qui supprime en douceur la croissance de cellules cancéreuses sans action de destruction des cellules, et qui empêche la récidive du cancer. L'invention concerne également un nouveau médicament anti-cancéreux. L'inhibiteur selon l'invention contient, comme composants efficaces : un peptide (A) contenant une séquence d'acides aminés représentée par SEQ ID NO. 1; ou un peptide (B) produit par suppression, remplacement ou ajout d'au moins un résidu d'acide aminé, dans une séquence d'acides aminés représentée par SEQ ID NO. 1, ledit peptide présentant une action de suppression de l'expression du gène de la cycline D1.
PCT/JP2014/054196 2013-02-21 2014-02-21 INHIBITEUR DE L'EXPRESSION DU GÈNE DE LA Cycline D1 ET MÉDICAMENT ANTICANCÉREUX ASSOCIÉ WO2014129590A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008056833A1 (fr) * 2006-11-10 2008-05-15 Livtech Inc. Anticorps anti-humain spécifiques de dlk1 présentant une activité anti-tumorale in vivo
WO2011040500A1 (fr) * 2009-09-29 2011-04-07 国立大学法人九州大学 Peptide inhibant la différenciation des cellules souches hématopoïétiques ou des cellules précurseurs hématopoïétiques et son utilisation
JP2012513775A (ja) * 2010-03-16 2012-06-21 コリア リサーチ インスティテュート オブ バイオサイエンス アンド バイオテクノロジー ＤＬＫ１−Ｆｃ融合タンパク質を有効成分として含む癌転移抑制用組成物

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2008056833A1 (fr) * 2006-11-10 2008-05-15 Livtech Inc. Anticorps anti-humain spécifiques de dlk1 présentant une activité anti-tumorale in vivo
WO2011040500A1 (fr) * 2009-09-29 2011-04-07 国立大学法人九州大学 Peptide inhibant la différenciation des cellules souches hématopoïétiques ou des cellules précurseurs hématopoïétiques et son utilisation
JP2012513775A (ja) * 2010-03-16 2012-06-21 コリア リサーチ インスティテュート オブ バイオサイエンス アンド バイオテクノロジー ＤＬＫ１−Ｆｃ融合タンパク質を有効成分として含む癌転移抑制用組成物

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