WO2018016797A1 - Composé pharmaceutique pour prévenir ou traiter un cancer résistant, récurrent ou métastatique. - Google Patents

Composé pharmaceutique pour prévenir ou traiter un cancer résistant, récurrent ou métastatique. Download PDF

Info

Publication number
WO2018016797A1
WO2018016797A1 PCT/KR2017/007519 KR2017007519W WO2018016797A1 WO 2018016797 A1 WO2018016797 A1 WO 2018016797A1 KR 2017007519 W KR2017007519 W KR 2017007519W WO 2018016797 A1 WO2018016797 A1 WO 2018016797A1
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
metastatic
resistant
pharmaceutical composition
emt
Prior art date
Application number
PCT/KR2017/007519
Other languages
English (en)
Korean (ko)
Inventor
김현석
정재호
Original Assignee
연세대학교 산학협력단
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 연세대학교 산학협력단 filed Critical 연세대학교 산학협력단
Publication of WO2018016797A1 publication Critical patent/WO2018016797A1/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
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/4545—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a six-membered ring with nitrogen as a ring hetero atom, e.g. pipamperone, anabasine
    • 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
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/4353—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/437—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having nitrogen as a ring hetero atom, e.g. indolizine, beta-carboline
    • 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
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4409—Non condensed pyridines; Hydrogenated derivatives thereof only substituted in position 4, e.g. isoniazid, iproniazid

Definitions

  • the present invention relates to a pharmaceutical composition for the prevention or treatment of resistant cancer, relapse cancer, or metastatic cancer, and more specifically, it can prevent or treat resistant cancer, relapse cancer, or metastatic cancer by using a Namtin (Nicotinamide phosphoribosyltransferase) inhibitor.
  • a pharmaceutical composition for the prevention or treatment of resistant cancer, relapse cancer, or metastatic cancer, and more specifically, it can prevent or treat resistant cancer, relapse cancer, or metastatic cancer by using a Namtin (Nicotinamide phosphoribosyltransferase) inhibitor.
  • Stomach cancer is one of the most common cancers worldwide, and it is the most common cancer occurring in Asia such as Korea and Japan.
  • the actual mortality rate shows a mortality rate of about 90% after deteriorating metastatic cancer rather than the state of primary tumor.
  • Herceptin which is used as an anticancer agent for HER2 + in breast cancer, is the only target anticancer agent currently used in metastatic gastric cancer, but it can be applied only to 16% of patients with metastatic gastric cancer. This exists.
  • Epithelial-mesenchymal transition is a process by which epithelial cells lose the characteristics of cell polarity and intercellular adhesion and obtain the mobility and invasive properties that can become mesenchymal stem cells. (Cancer and Metastasis Reviews 28, June 2009). When epithelial mesenchymal metastasis has occurred, the cells not only have characteristics such as mobility, invasiveness and stem cells, but also have a feature of preventing cell death and aging. Therefore, it has been reported that the cell diversity that may appear in the process can be confirmed in the early stages of metastasis of cancer (Cell 139, November 25, 2009).
  • CTCs circulating tumor cells
  • recurrence is one of the biggest challenges in cancer treatment. It is a common phenomenon in all chemotherapy and targeted chemotherapy, and can occur in both primary and metastatic cancers. Once relapsed, the prognosis is very poor. Epithelial mesenchymal metastasis is also known to cause cancer recurrence and resistance to anticancer agents. Cancer cells with epithelial mesenchymal metastases have been reported to induce cancer recurrence by inhibiting proliferation, increasing the expression of genes resistant to apoptosis and causing resistance to anticancer agents (Nature 527). , November 26, 2015).
  • An object of the present invention is to use a NamPT (Nicotinamide phosphoribosyltransferase) inhibitor to effectively inhibit the proliferation of resistant cancer, relapse cancer, or metastatic cancer cells to prevent and / or treat resistant cancer, relapse cancer, or metastatic cancer
  • the purpose is to provide.
  • the present invention administers a pharmaceutical composition comprising a Namtin (Nicotinamide phosphoribosyltransferase) inhibitor to an epithelial mesenchymal metastasis subtype (hereinafter, referred to as an 'EMT subtype') with the poorest prognosis of a patient among cancer cell lines
  • a Namtin Naturaltinamide phosphoribosyltransferase
  • an 'EMT subtype' epithelial mesenchymal metastasis subtype
  • EMT epithelial mesenchymal transition
  • the present invention relates to a pharmaceutical composition for preventing or treating resistant cancer, relapse cancer, or metastatic cancer, comprising a NamPT (Nicotinamide phosphoribosyltransferase) inhibitor as an active ingredient.
  • NamPT Natural-Nicotinamide phosphoribosyltransferase
  • the "NamPT inhibitor” means a drug that catalyzes the phosphoibosylation of nicotinamide and inhibits the activity of NamPT, an enzyme that mediates NAD biosynthesis.
  • the NamPT inhibitor is ((E) -N- [4- (1-benzoylpiperidin-4-yl) butyl] -3-pyridin-3-yl represented by the following formula (1) Prop-2-enamide) ((E) -N- [4- (1-benzoylpiperidin-4-yl) butyl] -3-pyridin-3-ylprop-2-enamide), N- (4-((3,5-difluorophenyl) sulfonyl) benzyl) imidazo [1,2-a] pyridine-6-carboxamide (N- (4-((3,5-difluorophenyl) sulfonyl ) benzyl) imidazo [1,2-a] pyridine-6-carboxamide) and (Z) -2- (6- (4-chlorophenoxy) hexyl) -1-cyano-3- ( Consisting of pyridine-4 (1H) -ylidine) gu
  • metalstatic cancer in the present invention means cancer caused by proliferation of cancer cells leaving other organs and moving to other organs.
  • the spread of cancer to other parts of the body can be largely divided into primary cancers in which cancerous tissues grow and directly invade surrounding organs and distant metastases along blood vessels or lymphatic vessels to other organs in the distance.
  • it may mean that the cancer has metastasized to the above organs in other primary cancers, but is not limited thereto.
  • the "resistant cancer” is extremely low sensitivity to cancer therapy or cancer therapy drugs, especially anticancer drugs, such as radiation therapy, so that the symptoms are not improved, alleviated, alleviated or treated by the therapy It does mean cancer.
  • the resistant cancer may be resistant to a specific treatment from the beginning or may not be initially resistant, but may be caused by a long time of treatment and no longer susceptible to the same treatment due to genetic mutations in cancer cells. .
  • the "recurrence cancer” means a case where the cancer occurs again at the site where the cancer occurred after receiving a cure determination after treatment of the first cancer.
  • the cancer may be a recurrence of cancer caused by epithelial mesenchymal metastasis process, but is not limited thereto.
  • the NamPT inhibitor may be applied without limitation as long as it is a cancer having resistance, recurrence, or metastasis, for example, breast cancer, biliary cancer, gallbladder cancer, pancreatic cancer, colon cancer, or uterine cancer having resistance, recurrence, or metastasis.
  • Esophageal cancer, stomach cancer, brain cancer, rectal cancer, lung cancer, bladder cancer, kidney cancer, ovarian cancer, prostate cancer, uterine cancer, head and neck cancer, skin cancer, hematologic cancer and liver cancer can be targeted to any one selected from cancer.
  • recurrent, or metastatic gastric cancer, colorectal cancer, pancreatic cancer, lung cancer, and breast cancer can be targeted to any one selected from cancer.
  • recurrent, or metastatic gastric cancer, colorectal cancer, pancreatic cancer, lung cancer, and breast cancer can be targeted to any one selected from cancer.
  • the resistant cancer, relapse cancer, or metastatic cancer may be an EMT subtype.
  • the metastatic cancer may be a cancer in which metastasis and / or relapse has occurred through an epithelial mesenchymal metastasis process, or a cancer that is difficult to treat due to resistance to drugs for treating cancer, and specific carcinoma is not particularly limited.
  • the cancer may be targeted to any one selected from cancer and liver cancer, and preferably, any one or more cancers selected from gastric cancer, colon cancer, pancreatic cancer, lung cancer and breast cancer of the EMT subtype.
  • the EMT subtype may be any one or more genes selected from the group consisting of SNAIL, E-cadherin, Vimentin, and CD44 or overexpressed or underexpressed proteins, preferably E.
  • the gene of -cadherin or the protein encoded by the gene may be underexpressed, or one or more genes selected from SNAIL, CD44 and Vimentin or the protein encoded by the gene may be overexpressed, but is not limited thereto. no.
  • the EMT subtype may be classified by a method for measuring the expression level of epidermal mesenchymal metastasis characteristic signature gene (BMC Medical Genomics 4, January 20, 2011).
  • the epithelial mesenchymal metastasis characteristic signature gene may be any one or more of over- and low-expression genes (Table 1 and 2).
  • the "signature score of epithelial mesenchymal metastasis” refers to the difference between the expression level average value of epithelial mesenchymal metastasis overexpression gene and the mean value of epithelial mesenchymal metastasis low expression gene in a subject.
  • the values can be used to classify the subject into an EMT subtype and a non-EMT subtype.
  • the epithelial mesenchymal metastasis characteristic overexpression genes are ADAM23, ADAMTS1, AFF3, AK5, AKAP12, ALPK2, ANGPTL2, ANKRD1, ANTXR1, ANXA6, AOX1, AP1S2, ARMCX1, ATP8B2, ATP8B3, AXL1, BDNFNC , BVES, FAM171A1, ZCCHC24, C16orf45, C1S, GLIPR2, CAP2, CCL2, CDH11, CDH2, CDH4, CHN1, CLDN11, CLIP3, CMTM3, COL12A1, COL1A2, COL3A1, COL4A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1, COL5A1,
  • epithelial mesenchymal metastasis-characteristic low expression genes are ACPP, AGR3, ALDH3B2, ANK3, ANKRD22, ANXA9, AP1M2, AQP3, ARHGAP8, ARHGDIB, PRR15L, ATP2C2, B3GNT3, BLNK, BSPRY, C11orf52f, MISP, F11f1 116f C1orf210, TTC39A, CCDC64B, CD24, CDH1, CDH3, CDS1, CEACAM5, CEACAM6, CGN, CKMT1B, CLDN4, CLDN7, CNKSR1, CNTNAP2, CTAGE4, DAPP1, DENND2D, DMKN, DSC2, EP3, EHF, ELF EPPK1, ERBB3, ERP27, FA2H, FAAH2, FAM110C, FAM83A, FAM84A, FAM84B, FBP1, FGD2, FGFBP1, FUT1, FUT3, FXYD3, G
  • the epithelial mesenchymal metastasis characteristic overexpression gene and low expression gene and its sequence number Entrez ID are shown in Table 1 and 2, respectively.
  • Overexpression genes Overexpression genes gene Entrez ID gene Entrez ID gene Entrez ID gene Entrez ID gene Entrez ID ADAM23 NM_8745 DPYSL3 NM_1809 NRG1 NM_3084 ADAMTS1 NM_9510 EDIL3 NM_10085 NUDT11 NM_55190 AFF3 NM_3899 ELOVL2 NM_54898 PAPPA NM_5069 AK5 NM_26289 EML1 NM_2009 PDE7B NM_27115 AKAP12 NM_9590 EMP3 NM_2014 PLAGL1 NM_5325 ALPK2 NM_115701 EPB41L5 NM_57669 PMP22 NM_5376 ANGPTL2 NM_23452 EPDR1 NM_54749 PNMA2 NM_10687 ANKRD1 NM_27063 EVI2A NM_2123 POPDC3 NM_64208 ANTXR1 NM_84168 F2R NM
  • a method for measuring the expression level of a gene overexpressed or underexpressed in the EMT subtype or a protein encoded by the gene is, for example, Western blot assay, ELISA (enzyme linked immunosorbent). assay, Radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, Complement Fixation Assay, FACS, RNA-sequencing Analysis, Microarray, RNA-scope, N-Counter, Reverse Phase Protein Array (RPPA) , FPPA (forward phase protein array), LC-MS / MS, ABCD (antibody barcoding with photocleavable DNA) and protein chips can be performed using, but not limited to.
  • Western blot assay ELISA (enzyme linked immunosorbent). assay, Radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immuno
  • the resistant cancer, relapse cancer, or metastatic cancer may be one in which the NAPRT (Nicotinic acid phosphoribosyltransferase) protein is down regulated.
  • the expression level of the protein may be evaluated to be low by comparing the expression level of the NAPRT with that of the normal control group.
  • the resistant cancer, recurrence cancer, or metastatic cancer may be included without limitation, especially among the EMT subtypes, in which the NAPRT protein is low in expression, for example, breast cancer, biliary cancer, gallbladder cancer, in which the NAPRT protein is low in expression,
  • the NAPRT protein may be gastric cancer, colon cancer, pancreatic cancer, lung cancer or breast cancer with low expression.
  • the "NAPRT protein” is an enzyme involved in the NAD biosynthesis process using nicotinic acid as a substrate in raw vegetables.
  • the biosynthesis of the NAD may be performed by de novo synthesis or by the salvage pathway, and the synthesis of the salvage pathway is performed by the nicotinic acid phosphoribosyltransferase (NAPRT) enzyme and nicotine.
  • NAPRT nicotinic acid phosphoribosyltransferase
  • NamPT Naturaltinamide phosphoribosyl transferase
  • amide nicotinamide
  • the resistant cancer, recurrence cancer, or metastatic cancer may be classified as a diffuse type gastric cancer by immunostaining.
  • the diffuse gastric cancer has a low NAPRT expression level, and thus, when the NamPT inhibitor according to the present invention is administered to the diffuse gastric cancer, NAD may inhibit biosynthesis of NAD, thereby inducing cell death more effectively.
  • the pharmaceutical composition of the present invention may be further administered in combination with other anticancer agents, thereby effectively inhibiting general cancer cell proliferation and cancer metastasis, thereby being used for the treatment of cancer.
  • the anticancer agent is nitrogen mustard, imatinib, oxaliplatin, rituximab, erlotinib, neratinib, lapatinib, zefitinib, vandetanib, nirotinib, semasanib, conservinib, axitinib, cediranib , Restautinib, trastuzumab, gefitinib, bortezomib, sunitinib, carboplatin, sorafenib, bevacizumab, cisplatin, cetuximab, biscumalboom, asparaginase, tretinoin, hydroxycarba Amide, dasatinib, estramastine, gemtuzumab ozogamycin, ibritumab tucetan, heptaplatin, methylaminolevulinic acid, amsacrine, alemtuzumab,
  • the pharmaceutical composition may be characterized in that the capsule, tablets, granules, injections, ointments, powder or beverage form, the pharmaceutical composition may be characterized in that it is intended for humans.
  • compositions of the present invention may be used in the form of oral dosage forms, such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.
  • the pharmaceutical composition of the present invention may comprise a pharmaceutically acceptable carrier.
  • Pharmaceutically acceptable carriers can be used as oral administration binders, suspending agents, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, pigments, fragrances, etc.
  • buffers, preservatives, analgesic Topical agents, solubilizers, isotonic agents, stabilizers and the like can be mixed and used, and for topical administration, bases, excipients, lubricants, preservatives and the like can be used.
  • the formulation of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with a pharmaceutically acceptable carrier as described above.
  • oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like, in the case of injections, in unit dosage ampoules or multiple dosage forms. have. And others, solutions, suspensions, tablets, capsules, sustained release preparations and the like.
  • suitable carriers, excipients and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil and the like can be used.
  • fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like may be further included.
  • Routes of administration of the pharmaceutical compositions according to the invention are not limited to these, but are oral, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, Sublingual or rectal. Oral or parenteral release is preferred.
  • parenteral includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intramuscular, intrasternal, intradural, intralesional and intracranial injection or infusion techniques.
  • the pharmaceutical compositions of the invention may also be administered in the form of suppositories for rectal administration.
  • the pharmaceutical compositions of the present invention vary depending on a number of factors including the activity, age, weight, general health, sex, formulation, time of administration, route of administration, rate of release, drug combination and severity of the particular disease to be prevented or treated, of the specific compound employed.
  • the dosage of the pharmaceutical composition may be appropriately selected by those skilled in the art depending on the patient's condition, weight, degree of disease, drug form, route of administration and duration, and 0.0001 to 50 mg / kg or It may be administered at 0.001 to 50 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
  • the pharmaceutical composition according to the present invention may be formulated as pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions.
  • the pharmaceutical composition according to the present invention can be used to prevent and / or treat cancer by effectively inducing cell proliferation and apoptosis among resistant cancer, recurrent cancer, or metastatic cancer cells, among cancers.
  • Figure 1 shows the results of EMT subtype gene characterization analysis according to an embodiment of the present invention.
  • Figure 2 shows the results of Western blot assay (Western blot assay) for confirming the expression of EMT-related proteins according to an embodiment of the present invention.
  • 3 to 5 show the results of Matrigel (Matrigel) invasion analysis according to an embodiment of the present invention.
  • Figure 20 shows the results of EMT subtype cell line survival analysis according to an embodiment of the present invention.
  • Figure 21 shows the result of Western blot assay (Western blot assay) for confirming NAPRT protein expression according to an embodiment of the present invention.
  • 25 to 38 show the results of NAPRT gene expression in EMT and non-EMT subtypes and normal tissues of five carcinomas according to an embodiment of the present invention.
  • the pharmaceutical composition comprising NamPT (Nicotinamide phosphoribosyltransferase) inhibitor of the present invention as an active ingredient is a cell proliferation and cells of resistant cancer, recurrent cancer, or metastatic cancer cells having the characteristics of epithelial-mesenchymal transition (EMT) subtype. It can be effectively used to prevent and / or treat cancer by effectively inducing death.
  • NamPT Natural-Nicotinamide phosphoribosyltransferase
  • EMT epithelial-mesenchymal transition
  • ((E) -N- [4- (1-benzoylpiperidin-4-yl) butyl] -3-pyridin-3-ylprop-2-enamide) according to the invention
  • FBS fetal bovine serum
  • antibiotics penicillin-streptomycin (Invitrogen)
  • EMT epithelium Mesenchymal metastasis
  • the genes used in the classification of gastric cancer cohort EMT subtypes of ACRG were up-regulated and down-regulated 161 genes. Unsupervised hierarchical clustering was performed.
  • the epithelial mesenchymal transition gene feature autonomic hierarchical clustering analysis method is as follows. For gastric cancer cell lines, RNA-sequence analysis was used to calculate normalized expression levels for the transcripts of 28 cell lines. 1 was added to the FPKM value obtained by the expression analysis and then converted to the log2 level was used for analysis. The R package hclust was used to perform autonomic hierarchical clustering with the expression values of the 149 overexpression genes and 161 low expression genes. The distance was calculated using the maximum linkage by Euclidean method, and the results are shown in FIG. 1.
  • Hs746T, SNU484, YCC11, SNU1750, SK4, SNU668 and MKN1 cell lines belong to the EMT subtype.
  • Epithelial mesenchymal transition marker protein expression was confirmed by Western blot assay to verify the epithelial mesenchymal metastasis characteristics of the EMT subtype cell line.
  • the preparation method of the protein used for the Western blot analysis is as follows.
  • the biliary cancer cell line was harvested by treatment with PBS, washed with PBS, and lysed using RIPA buffer, a cell lysis buffer containing proteolytic inhibitor.
  • the dissolved solution was fractionated from the total solution using a centrifuge, and only the solution containing the protein was extracted.
  • the extracted protein was quantified by the Bradford method.
  • the same concentration of protein obtained through quantification was separated by SDS-polyacrylamide gel electrophoresis and transferred to PVDF membrane.
  • the membrane to which the protein was transferred was blocked with Tris-buffered saline / 0.1% Tween-20 (TBS-T) solution containing 5% non-fat milk for 1 hour at room temperature to reduce nonspecific binding.
  • TBS-T Tris-buffered saline / 0.1% Tween-20
  • the primary antibody diluted in 5% BSA solution was reacted at 4 ° C. for at least 12 hours, and the secondary antibody (1: 5000 dilution) was reacted at room temperature for 1 hour.
  • an enhanced chemiluminescence system was used for visualization.
  • E-cadherin expression is low in epithelial mesenchymal metastasis cell lines, and although there is a difference in the degree of expression, expression of Vimentin and snail does not occur epithelial mesenchymal metastasis without epithelial mesenchymal metastasis. (Hereinafter referred to as 'non-EMT') It was confirmed that the increase compared to the cell line. On the other hand, in the case of the non-EMT cell line, the epithelial mesenchymal metastasis cell line and the expression pattern was found to be reversed.
  • Matrigel invasion assay method was cultured in serum-free medium epithelial mesenchymal transition cell line and control in a transwell coated with Matrigel at a concentration of 300 ⁇ g / ml.
  • the lower part of the transwell was a nutrient medium containing fetal calf serum (FBS) and cultured in an incubator at 37 ° C., 5%, and CO 2 .
  • FBS fetal calf serum
  • MKN45 and NCC59 cells which are cell lines without epithelial mesenchymal metastasis, were used.
  • the matrigel membrane were fixed with methanol, and stained with 0.2% crystal violet.
  • the number of cells contained in the membrane was counted, and the results are shown in FIGS. 3 to 5.
  • the sphere formation experiment was carried out by incubating the epithelial mesenchymal metastatic subtype cell line and control group MKN45 in DMEM-F12 medium (20 ng / ml EGF, 20 ng / ml basal FGF, 2% B27 supplement) without serum. was observed under a microscope. Moreover, the number of the cells which formed the sphere was aggregated, and the result is shown to FIGS. 6-8.
  • EMT subtype cell lines SK4, Hs746T, SNU484, SNU1750, YCC11 and SNU668 according to the present invention have cancer stem cell properties.
  • the small molecule pharmacological compounds are related to oncology, cardiology, anti-inflammatory, immunology, neuropsychiatry and anticancer, and the anticancer small molecule compounds are known to have anticancer effects and have been used in preclinical, clinical trial stages or tested. All materials were purchased from Selleckchem.
  • each of the seven EMT subtype gastric cancer cell lines according to the present invention was dispensed at 5,000 wells in 96 well-plates, and the drug concentration in the cell culture solution was diluted to 2.5 ⁇ M at 24 hours.
  • Cells were added using a Mac Liquid Handler instrument (Beckman).
  • the control group was treated with the same concentration of DMSO as the experimental group.
  • the cells were incubated for 72 hours in a 37 ° C., 5% CO 2 incubator, and then cell viability was measured using a CellTiter-Glo Assay Kit (Promega), and the results are shown in FIGS. 9 to 15.
  • the viability values were divided by 0.1 units, and the number of drugs selected according to the number of cell lines corresponding to the corresponding viability values or less is shown in FIGS. 16 and 17.
  • the 63 small molecule compounds include 9 drugs related to DNA synthesis, Topoisomerase, 6 cell cycle inhibitors, 8 HDAC inhibitors, 3 EGFR, HER2, PIK3CA and src related drugs, 4 mTOR and PI3K inhibitors, and proteasome 4 related drugs, 2 autophagy related drugs, 5 microtubule related cases, 2 DNA / RNA damage related cases, 2 ALK related cases, 3 Ubiquitine ligase cases, 6 infection related cases and 3 other cases were identified (Table 3). .
  • Target Drug Topoisomerase DNA synthesis Epirubicin Hydrochloride, Mitoxantrone Hydrochloride, Camptothecin, Doxorubicin (Adriamycin), Gemcitabine, Daunorubicin HCl (Daunocycine HCl), Gemcitabine HCl (Gemzar), Clofarabine, Niclosamide (Niclocide), Fludarabine Phosphate (Fludara) Cell cycle (Aurora, cdk plk1, etc) Danusertib (PHA-739358), AT9283, Flavopiridol (Alvocidib) HCl, BI2536, GSK461364, BI6727 (Volasertib), SB743921, Ispinesib (SB-715992) HDAC Trichostatin A (TSA), PCI-24781, CUDC-101, JNJ-26481585, Belinostat (PXD101), SB939 (Paracino
  • the 29 gastric cancer cell lines were dispensed in 5,000 wells each into 96 well-plates, and the drug concentration in the cell culture solution was measured by log serial dilution at 24 hours. Dilutions were made at 12 concentrations corresponding to a minimum of 0.5 nM to a maximum of 50 ⁇ M and added to the cells using a BioMac Liquid Handler instrument (Beckman). The cells were incubated for 72 hours in a 37 ° C., 5% CO 2 incubator, and cell viability was measured using a CellTiter-Glo assay kit (Promega). The measured values yielded cell viability values for 12 concentrations per drug for each of 63 drugs and 29 cell lines.
  • the AUC (Area under the curve) value which is an indicator of cell line specific sensitivity for 63 drugs, was calculated, and the results are shown in FIG. 18. Also, to classify the 63 drugs based on the difference in AUC values between EMT subtypes and non-EMT subtypes, the difference between the median value of the AUC values of 7 EMT gastric cancer cell lines and the AUC median value of 22 non-EMT cell lines. was calculated. In addition, 63 drugs were classified based on the p-value obtained by performing the Student t-test to confirm the significant difference between the two groups, and the results are shown in FIG. 19.
  • the drug with low AUC blue was found to have high drug sensitivity in the cell line.
  • FK866 showed the largest AUC difference between gastric cancer cells of the EMT subtype and gastric cancer cell lines of the non-EMT subtype and had a significant p-value.
  • the effect of inhibiting survival rate by FK866 was confirmed in the EMT subtype cell line. After incubating 72 hours with 0-50 ⁇ M of FK866 to each of the cell lines, the survival rate was measured in the manner of Example 3, and the results are shown in FIG. 20.
  • FK866 according to the present invention has therapeutic efficacy in gastric cancer cell lines having EMT subtypes.
  • NAPRT As shown in FIG. 21, it was confirmed that the expression of NAPRT was reduced in five cells except YCC1 and SK4 classified as EMT subtypes. SK4 cell lines with high NAPRT expression were exceptionally resistant to FK866. In addition, YCC1 showed low sensitivity to FK866 for NAPRT expression. On the other hand, seven cell lines belonging to the non-EMT was confirmed that the expression of NAPRT at a high level.
  • the total NAD + / NADH amount was analyzed in the cell lines SNU668 and Hs746T and non-EMT cell lines of the EMT subtype.
  • the NAD protein amount analysis was quantitatively measured NAD + / NADH using Ultra-Glo TM Recombinant Luciferase assay kit (Promega, # G9071). Specifically, 5000 cells were cultured in 96-well plates, and then FK866 was administered at a concentration of 2.5-20 nM, followed by a culture process for 40 hours. After the above process, after treating the cyclic enzyme converting NAD + to NADH, the activity of reductase by NADH was measured, and the results are shown in FIG. 22.
  • Each of the EMT subtype cell lines was treated with 25 nM FK866, and nicotinic acid (NA), nicotinamide (NAM) and NAD were added to the cell culture medium at 50 to 200 mM to measure survival. The results are shown in FIGS. 23 and 24.
  • Analyzes were performed to classify samples corresponding to EMT subtypes from published transcript datasets in a number of carcinomas. Then, the amount of expression of NAPRT between the EMT subtype and the non-EMT subtype in the sorted samples was compared.
  • NAPRT expression levels were performed by TCGA, Gene Expression Omnibus (GEO) for 17 carcinomas [Barrett et al. 2013 PMID 23193258] and ArrayExpress [Kolesnikov et al. 2015 PMID 25361974] utilized transcript profiles of 33 cohorts collected from ttps: //tcga-data.nci.nih.gov/tcga/, http: Downloaded from the web link at //www.ncbi.nlm.nih.gov/geo/, https://www.ebi.ac.uk/arrayexpress and used for analysis.
  • GEO Gene Expression Omnibus
  • the E-MTAB-923 data set was normalized to a robust multichip average (RMA) method using affy R package, and then converted to Log2 level for analysis.
  • the RNA-seq data set was used for the E-MTAB-923 data set.
  • RSEM method when normalized by the above method, it was normalized by the RSEM method.
  • 1 was added to the normalized RPKM value and then converted to the log2 level to be used for the following analysis.
  • Samples belonging to the EMT subtype are calculated from the average value of the low-expression 161 genes from the average value of the 149 genes overexpressing the EMT subtypes, and then displayed in a quantile-quartile gram (q-q plot) through the difference values.
  • q-q plot the point which begins to deviate in the direction of the high score from the diagonal distribution trend connecting the first quantile and the third quantile is determined as the threshold, and the epithelium is above the threshold. If the mesenchymal metastasis gene feature scores were large, they were classified as EMT subtypes.
  • the assay was used for analysis only if EMT subtypes were identified in at least 3% of the total samples per each cohort.
  • the expression level of NAPRT gene between the EMT subtype and non-EMT subtype samples in gastric cancer, colorectal cancer, pancreatic cancer, lung adenocarcinoma and breast cancer, where more than 3% of EMT subtypes were found in at least two cohorts. are shown in FIGS. 25 to 38 and 39.
  • EMT subtypes are present not only in gastric cancer but also in colorectal cancer, pancreatic cancer, lung cancer and breast cancer, and NAPRT low expression is observed in cancers of all the EMT subtypes. It has been found to be useful as a target anticancer drug for cancer.
  • TMA tissue microarray
  • NamPT inhibitor including FK866 of the present invention will induce apoptosis by interfering with the biosynthesis of NAD in subtype gastric cancer with low NAPRT expression.
  • the pharmaceutical composition comprising the NamPT (Nicotinamide phosphoribosyltransferase) inhibitor of the present invention as an active ingredient can be effectively used to prevent and / or treat cancer by effectively inducing cell proliferation and apoptosis of resistant cancer, relapse cancer, or metastatic cancer cells. have.
  • NamPT Natural-Nicotinamide phosphoribosyltransferase

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

L'invention porte sur un composé pharmaceutique destiné à prévenir ou à traiter un cancer résistant, récurrent ou métastatique à l'aide d'un inhibiteur de nicotinamide phosphoribosyltransférase (NAMPT). Cette invention peut être utilisée pour prévenir et/ou traiter le cancer en provoquant la croissance et la mort des cellules d'un cancer résistant, récurrent ou métastatique.
PCT/KR2017/007519 2016-07-20 2017-07-13 Composé pharmaceutique pour prévenir ou traiter un cancer résistant, récurrent ou métastatique. WO2018016797A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020160092133A KR101869794B1 (ko) 2016-07-20 2016-07-20 내성암, 재발암 또는 전이암의 예방 또는 치료용 약학 조성물
KR10-2016-0092133 2016-07-20

Publications (1)

Publication Number Publication Date
WO2018016797A1 true WO2018016797A1 (fr) 2018-01-25

Family

ID=60993213

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2017/007519 WO2018016797A1 (fr) 2016-07-20 2017-07-13 Composé pharmaceutique pour prévenir ou traiter un cancer résistant, récurrent ou métastatique.

Country Status (2)

Country Link
KR (1) KR101869794B1 (fr)
WO (1) WO2018016797A1 (fr)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130114119A (ko) * 2010-09-03 2013-10-16 포르마 티엠, 엘엘씨. Ｎａｍｐｔ의 억제를 위한 신규 화합물 및 조성물
KR20130114653A (ko) * 2010-09-03 2013-10-17 포르마 티엠, 엘엘씨. 암과 같은 질환의 치료를 위한 ｎａｍｐｔ 억제제로서의 4-｛［（피리딘-3-일-메틸）아미노카르보닐］아미노｝벤젠-술폰 유도체
KR20140020823A (ko) * 2010-09-03 2014-02-19 포르마 티엠, 엘엘씨. Ｎａｍｐｔ의 억제를 위한 구아니딘 화합물 및 조성물
JP2015522028A (ja) * 2012-06-27 2015-08-03 アルツハイマーズ・インスティテュート・オブ・アメリカ・インコーポレイテッドＡｌｚｈｅｉｍｅｒ’Ｓ Ｉｎｓｔｉｔｕｔｅ Ｏｆ Ａｍｅｒｉｃａ， Ｉｎｃ． 化合物とその治療用途

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20130114119A (ko) * 2010-09-03 2013-10-16 포르마 티엠, 엘엘씨. Ｎａｍｐｔ의 억제를 위한 신규 화합물 및 조성물
KR20130114653A (ko) * 2010-09-03 2013-10-17 포르마 티엠, 엘엘씨. 암과 같은 질환의 치료를 위한 ｎａｍｐｔ 억제제로서의 4-｛［（피리딘-3-일-메틸）아미노카르보닐］아미노｝벤젠-술폰 유도체
KR20140020823A (ko) * 2010-09-03 2014-02-19 포르마 티엠, 엘엘씨. Ｎａｍｐｔ의 억제를 위한 구아니딘 화합물 및 조성물
JP2015522028A (ja) * 2012-06-27 2015-08-03 アルツハイマーズ・インスティテュート・オブ・アメリカ・インコーポレイテッドＡｌｚｈｅｉｍｅｒ’Ｓ Ｉｎｓｔｉｔｕｔｅ Ｏｆ Ａｍｅｒｉｃａ， Ｉｎｃ． 化合物とその治療用途

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BI, T.-Q.: "Overexpression of Nampt in gastric cancer and chemopotentiating effects of the Nampt inhibitor FK866 in combination with fluorouracil", ONCOLOGY REPORTS, vol. 26, no. 5, 2011, pages 1251 - 1257, XP055602702, DOI: 10.3892/or.2011.1378 *

Also Published As

Publication number Publication date
KR101869794B1 (ko) 2018-06-21
KR20180010046A (ko) 2018-01-30

Similar Documents

Publication Publication Date Title
JP2019131546A (ja) がん患者におけるがんの治療方法及び薬物応答性の予測方法
TW201609107A (zh) 布魯頓氏酪胺酸激酶（ｂｒｕｔｏｎ’ｓ ｔｙｒｏｓｉｎｅ ｋｉｎａｓｅ ）抑制劑之組合及其用途
Ling et al. Aberrant methylation of different DNA repair genes demonstrates distinct prognostic value for esophageal cancer
EP2655661B1 (fr) Procédés et moyens de classification moléculaire des cancers colorectaux
WO2021241981A1 (fr) Composition pour la prévention ou le traitement du cancer
JP2022543555A (ja) ｍｉＲＮＡ－１９３ａに関する新たな処置
WO2019081507A1 (fr) Nouveaux marqueurs dérivés du sang pour la détection du cancer
US20160058777A1 (en) Identification and treatment of cancer subsets
US11913959B2 (en) Methods of treating and prognosing nonhematopoietic malignant tumors
WO2018016797A1 (fr) Composé pharmaceutique pour prévenir ou traiter un cancer résistant, récurrent ou métastatique.
US20180298447A1 (en) Methods for assessing the treatment response of cancer patients and for treating cancer patients by analysing cpg methylation
KR20190085903A (ko) 내성암, 재발암 또는 전이암의 예방 또는 치료용 약학 조성물
US20190388453A1 (en) Treatment of drug resistant proliferative diseases with telomerase mediated telomere altering compounds
KR101937126B1 (ko) 내성암, 재발암 또는 전이암의 예방 또는 치료용 약학 조성물
WO2019083458A1 (fr) Varlitinib s'utilisant dans le traitement du cancer pour réduire l'hypoxie
WO2019083456A1 (fr) Varlitinib destinée à être utilisé dans le traitement du cancer pour normaliser l'angiogenèse dans une masse cancéreuse
WO2019083457A1 (fr) Varlitinib destiné à être utilisé dans le traitement du cancer chez un patient identifié comme ayant une mutation de la voie de la bêta-caténine
KR20200022187A (ko) Nono의 발현 또는 활성 억제제를 유효성분으로 포함하는 방사선 민감성 증진용 조성물
WO2021080363A1 (fr) Composition pour la prévention ou le traitement du cancer
WO2023083308A1 (fr) Procédé d'évaluation du pronostic du cancer colorectal reposant sur la méthylation de l'adn
US20230257825A1 (en) Breast cancer biomarkers and methods of use
Yang Identification of genes and pathways involved in pancreatic adenocarcinoma using bioinformatics and experiment analysis
Aggarwal et al. Therapy-related cutaneous adverse effects in an Indian cohort of adult leukaemia patients.
Arrait The Study of Pharmacogenomic Effect of Capicitabine in Colorectal Cancer Patients
Wosiak et al. Assessment of the Influence of 5-fluorouracil on the SMAD4 Gene Expression, Apoptosis induction and DNA Damage in the Human CACO-2 Cell Line

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: 17831270

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17831270

Country of ref document: EP

Kind code of ref document: A1