KR20220127570A - Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same - Google Patents
Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same Download PDFInfo
- Publication number
- KR20220127570A KR20220127570A KR1020210032023A KR20210032023A KR20220127570A KR 20220127570 A KR20220127570 A KR 20220127570A KR 1020210032023 A KR1020210032023 A KR 1020210032023A KR 20210032023 A KR20210032023 A KR 20210032023A KR 20220127570 A KR20220127570 A KR 20220127570A
- Authority
- KR
- South Korea
- Prior art keywords
- cancer
- composition
- present
- cells
- compound
- Prior art date
Links
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 50
- 201000011510 cancer Diseases 0.000 title claims abstract description 42
- 239000000203 mixture Substances 0.000 title claims abstract description 37
- 125000003406 indolizinyl group Chemical class C=1(C=CN2C=CC=CC12)* 0.000 title abstract 2
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims abstract description 36
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 claims abstract description 36
- 201000002528 pancreatic cancer Diseases 0.000 claims abstract description 36
- 208000008443 pancreatic carcinoma Diseases 0.000 claims abstract description 36
- 230000000694 effects Effects 0.000 claims abstract description 27
- 150000001875 compounds Chemical class 0.000 claims abstract description 23
- 102100021948 Lysyl oxidase homolog 2 Human genes 0.000 claims description 30
- 150000003839 salts Chemical class 0.000 claims description 18
- 230000002401 inhibitory effect Effects 0.000 claims description 16
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 11
- 108010055717 JNK Mitogen-Activated Protein Kinases Proteins 0.000 claims description 11
- 239000001257 hydrogen Substances 0.000 claims description 11
- 229910052739 hydrogen Inorganic materials 0.000 claims description 11
- 239000004480 active ingredient Substances 0.000 claims description 10
- 125000000217 alkyl group Chemical group 0.000 claims description 6
- 235000013376 functional food Nutrition 0.000 claims description 4
- 101001043352 Homo sapiens Lysyl oxidase homolog 2 Proteins 0.000 claims description 3
- 230000006872 improvement Effects 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 206010006187 Breast cancer Diseases 0.000 claims description 2
- 208000026310 Breast neoplasm Diseases 0.000 claims description 2
- 206010009944 Colon cancer Diseases 0.000 claims description 2
- 208000001333 Colorectal Neoplasms Diseases 0.000 claims description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 claims description 2
- 206010017758 gastric cancer Diseases 0.000 claims description 2
- 201000011549 stomach cancer Diseases 0.000 claims description 2
- 206010038111 Recurrent cancer Diseases 0.000 claims 1
- 230000001394 metastastic effect Effects 0.000 claims 1
- 208000037819 metastatic cancer Diseases 0.000 claims 1
- 208000011575 metastatic malignant neoplasm Diseases 0.000 claims 1
- 230000035755 proliferation Effects 0.000 abstract description 11
- 206010027476 Metastases Diseases 0.000 abstract description 10
- 230000009401 metastasis Effects 0.000 abstract description 10
- 230000022131 cell cycle Effects 0.000 abstract description 8
- 238000011161 development Methods 0.000 abstract description 8
- 102000004190 Enzymes Human genes 0.000 abstract description 7
- 108090000790 Enzymes Proteins 0.000 abstract description 7
- 102000029749 Microtubule Human genes 0.000 abstract description 5
- 108091022875 Microtubule Proteins 0.000 abstract description 5
- -1 indolizine derivative compound Chemical class 0.000 abstract description 5
- 210000004688 microtubule Anatomy 0.000 abstract description 5
- 239000002246 antineoplastic agent Substances 0.000 abstract description 3
- 238000006116 polymerization reaction Methods 0.000 abstract description 3
- 230000011278 mitosis Effects 0.000 abstract description 2
- 210000004881 tumor cell Anatomy 0.000 abstract description 2
- 229940041181 antineoplastic drug Drugs 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 63
- 101710183215 Lysyl oxidase homolog 2 Proteins 0.000 description 27
- 239000003814 drug Substances 0.000 description 15
- 108090000623 proteins and genes Proteins 0.000 description 15
- 230000006907 apoptotic process Effects 0.000 description 14
- 102000004169 proteins and genes Human genes 0.000 description 13
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 11
- 229940079593 drug Drugs 0.000 description 10
- 239000008194 pharmaceutical composition Substances 0.000 description 10
- 201000010099 disease Diseases 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 230000005754 cellular signaling Effects 0.000 description 8
- 230000018109 developmental process Effects 0.000 description 7
- 238000000684 flow cytometry Methods 0.000 description 7
- 235000013305 food Nutrition 0.000 description 7
- 229960005277 gemcitabine Drugs 0.000 description 7
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000001093 anti-cancer Effects 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 6
- 230000001225 therapeutic effect Effects 0.000 description 6
- 238000001262 western blot Methods 0.000 description 6
- 102000004243 Tubulin Human genes 0.000 description 5
- 108090000704 Tubulin Proteins 0.000 description 5
- 230000004663 cell proliferation Effects 0.000 description 5
- 239000012091 fetal bovine serum Substances 0.000 description 5
- 239000000796 flavoring agent Substances 0.000 description 5
- 235000013355 food flavoring agent Nutrition 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- MQLACMBJVPINKE-UHFFFAOYSA-N 10-[(3-hydroxy-4-methoxyphenyl)methylidene]anthracen-9-one Chemical compound C1=C(O)C(OC)=CC=C1C=C1C2=CC=CC=C2C(=O)C2=CC=CC=C21 MQLACMBJVPINKE-UHFFFAOYSA-N 0.000 description 4
- 108090000854 Oxidoreductases Proteins 0.000 description 4
- 102000004316 Oxidoreductases Human genes 0.000 description 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 238000010609 cell counting kit-8 assay Methods 0.000 description 4
- 239000003937 drug carrier Substances 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 150000002478 indolizines Chemical class 0.000 description 4
- 239000003112 inhibitor Substances 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 230000000394 mitotic effect Effects 0.000 description 4
- 229940124597 therapeutic agent Drugs 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- QAPSNMNOIOSXSQ-YNEHKIRRSA-N 1-[(2r,4s,5r)-4-[tert-butyl(dimethyl)silyl]oxy-5-(hydroxymethyl)oxolan-2-yl]-5-methylpyrimidine-2,4-dione Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O[Si](C)(C)C(C)(C)C)C1 QAPSNMNOIOSXSQ-YNEHKIRRSA-N 0.000 description 3
- YXHLJMWYDTXDHS-IRFLANFNSA-N 7-aminoactinomycin D Chemical compound C[C@H]1OC(=O)[C@H](C(C)C)N(C)C(=O)CN(C)C(=O)[C@@H]2CCCN2C(=O)[C@@H](C(C)C)NC(=O)[C@H]1NC(=O)C1=C(N)C(=O)C(C)=C2OC(C(C)=C(N)C=C3C(=O)N[C@@H]4C(=O)N[C@@H](C(N5CCC[C@H]5C(=O)N(C)CC(=O)N(C)[C@@H](C(C)C)C(=O)O[C@@H]4C)=O)C(C)C)=C3N=C21 YXHLJMWYDTXDHS-IRFLANFNSA-N 0.000 description 3
- 108700012813 7-aminoactinomycin D Proteins 0.000 description 3
- 102000004121 Annexin A5 Human genes 0.000 description 3
- 108090000672 Annexin A5 Proteins 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- KYRVNWMVYQXFEU-UHFFFAOYSA-N Nocodazole Chemical compound C1=C2NC(NC(=O)OC)=NC2=CC=C1C(=O)C1=CC=CS1 KYRVNWMVYQXFEU-UHFFFAOYSA-N 0.000 description 3
- 229920000776 Poly(Adenosine diphosphate-ribose) polymerase Polymers 0.000 description 3
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 230000005757 colony formation Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 230000007705 epithelial mesenchymal transition Effects 0.000 description 3
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 238000003125 immunofluorescent labeling Methods 0.000 description 3
- 229950006344 nocodazole Drugs 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 2
- FQNGLPGFBJLTJM-UHFFFAOYSA-N 6-amino-5-(4-methoxybenzoyl)indolizine-7-carbonitrile Chemical compound C1=CC(OC)=CC=C1C(=O)C1=C(N)C(C#N)=CC2=CC=CN12 FQNGLPGFBJLTJM-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- 101150012716 CDK1 gene Proteins 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000003952 Caspase 3 Human genes 0.000 description 2
- 108090000397 Caspase 3 Proteins 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- 229930091371 Fructose Natural products 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 2
- 101000911513 Homo sapiens Uncharacterized protein FAM215A Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 101710179684 Poly [ADP-ribose] polymerase Proteins 0.000 description 2
- 102100023712 Poly [ADP-ribose] polymerase 1 Human genes 0.000 description 2
- 108010087230 Sincalide Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 2
- 102100026728 Uncharacterized protein FAM215A Human genes 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 235000011054 acetic acid Nutrition 0.000 description 2
- 230000002424 anti-apoptotic effect Effects 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 239000002585 base Substances 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 230000004611 cancer cell death Effects 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 230000006369 cell cycle progression Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000010293 colony formation assay Methods 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 208000035475 disorder Diseases 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 229960002949 fluorouracil Drugs 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000009545 invasion Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 230000008880 microtubule cytoskeleton organization Effects 0.000 description 2
- 230000025090 microtubule depolymerization Effects 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 238000010837 poor prognosis Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- ZSKGQVFRTSEPJT-UHFFFAOYSA-N pyrrole-2-carboxaldehyde Chemical compound O=CC1=CC=CN1 ZSKGQVFRTSEPJT-UHFFFAOYSA-N 0.000 description 2
- HELXLJCILKEWJH-NCGAPWICSA-N rebaudioside A Chemical compound O([C@H]1[C@H](O)[C@@H](CO)O[C@H]([C@@H]1O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)O[C@]12C(=C)C[C@@]3(C1)CC[C@@H]1[C@@](C)(CCC[C@]1([C@@H]3CC2)C)C(=O)O[C@H]1[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O1)O)[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O HELXLJCILKEWJH-NCGAPWICSA-N 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 235000010356 sorbitol Nutrition 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 239000003656 tris buffered saline Substances 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- DGVVWUTYPXICAM-UHFFFAOYSA-N β‐Mercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 2
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 1
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- XQJAHBHCLXUGEP-UHFFFAOYSA-N 2-bromo-1-(4-methoxyphenyl)ethanone Chemical compound COC1=CC=C(C(=O)CBr)C=C1 XQJAHBHCLXUGEP-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241001479434 Agfa Species 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 108010063104 Apoptosis Regulatory Proteins Proteins 0.000 description 1
- 102000010565 Apoptosis Regulatory Proteins Human genes 0.000 description 1
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- QFGHXZSVKSDWGB-UHFFFAOYSA-N COC1=CC=C(C=C1)C(CN1C(=CC=C1)C=O)=O Chemical compound COC1=CC=C(C=C1)C(CN1C(=CC=C1)C=O)=O QFGHXZSVKSDWGB-UHFFFAOYSA-N 0.000 description 1
- 241000282472 Canis lupus familiaris Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000700198 Cavia Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 238000000116 DAPI staining Methods 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 230000006820 DNA synthesis Effects 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 239000004278 EU approved seasoning Substances 0.000 description 1
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 239000004386 Erythritol Substances 0.000 description 1
- UNXHWFMMPAWVPI-UHFFFAOYSA-N Erythritol Natural products OCC(O)C(O)CO UNXHWFMMPAWVPI-UHFFFAOYSA-N 0.000 description 1
- 229940124602 FDA-approved drug Drugs 0.000 description 1
- 239000001512 FEMA 4601 Substances 0.000 description 1
- 241000282326 Felis catus Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 208000034826 Genetic Predisposition to Disease Diseases 0.000 description 1
- 239000004378 Glycyrrhizin Substances 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 241000282560 Macaca mulatta Species 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 239000007990 PIPES buffer Substances 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 241000282579 Pan Species 0.000 description 1
- 241000282520 Papio Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 108091026813 Poly(ADPribose) Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 239000012083 RIPA buffer Substances 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- HELXLJCILKEWJH-SEAGSNCFSA-N Rebaudioside A Natural products O=C(O[C@H]1[C@@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1)[C@@]1(C)[C@@H]2[C@](C)([C@H]3[C@@]4(CC(=C)[C@@](O[C@H]5[C@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@@H](O[C@H]6[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O6)[C@H](O)[C@@H](CO)O5)(C4)CC3)CC2)CCC1 HELXLJCILKEWJH-SEAGSNCFSA-N 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 239000012722 SDS sample buffer Substances 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 244000228451 Stevia rebaudiana Species 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 244000126002 Ziziphus vulgaris Species 0.000 description 1
- 239000000205 acacia gum Substances 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 150000001340 alkali metals Chemical class 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 150000001342 alkaline earth metals Chemical class 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000001668 ameliorated effect Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- SRSXLGNVWSONIS-UHFFFAOYSA-N benzenesulfonic acid Chemical compound OS(=O)(=O)C1=CC=CC=C1 SRSXLGNVWSONIS-UHFFFAOYSA-N 0.000 description 1
- 229940092714 benzenesulfonic acid Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 229960001714 calcium phosphate Drugs 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 229960003340 calcium silicate Drugs 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000032823 cell division Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 238000001437 electrospray ionisation time-of-flight quadrupole detection Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- HELXLJCILKEWJH-UHFFFAOYSA-N entered according to Sigma 01432 Natural products C1CC2C3(C)CCCC(C)(C(=O)OC4C(C(O)C(O)C(CO)O4)O)C3CCC2(C2)CC(=C)C21OC(C1OC2C(C(O)C(O)C(CO)O2)O)OC(CO)C(O)C1OC1OC(CO)C(O)C(O)C1O HELXLJCILKEWJH-UHFFFAOYSA-N 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 235000019414 erythritol Nutrition 0.000 description 1
- UNXHWFMMPAWVPI-ZXZARUISSA-N erythritol Chemical compound OC[C@H](O)[C@H](O)CO UNXHWFMMPAWVPI-ZXZARUISSA-N 0.000 description 1
- 229940009714 erythritol Drugs 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- DEFVIWRASFVYLL-UHFFFAOYSA-N ethylene glycol bis(2-aminoethyl)tetraacetic acid Chemical compound OC(=O)CN(CC(O)=O)CCOCCOCCN(CC(O)=O)CC(O)=O DEFVIWRASFVYLL-UHFFFAOYSA-N 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 235000013312 flour Nutrition 0.000 description 1
- 235000011194 food seasoning agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 235000015203 fruit juice Nutrition 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 229940014259 gelatin Drugs 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 235000001727 glucose Nutrition 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- LPLVUJXQOOQHMX-UHFFFAOYSA-N glycyrrhetinic acid glycoside Natural products C1CC(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2=O)C(O)=O)C)(C)CC2)(C)C2C(C)(C)C1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O LPLVUJXQOOQHMX-UHFFFAOYSA-N 0.000 description 1
- 239000001649 glycyrrhiza glabra l. absolute Substances 0.000 description 1
- 229960004949 glycyrrhizic acid Drugs 0.000 description 1
- UYRUBYNTXSDKQT-UHFFFAOYSA-N glycyrrhizic acid Natural products CC1(C)C(CCC2(C)C1CCC3(C)C2C(=O)C=C4C5CC(C)(CCC5(C)CCC34C)C(=O)O)OC6OC(C(O)C(O)C6OC7OC(O)C(O)C(O)C7C(=O)O)C(=O)O UYRUBYNTXSDKQT-UHFFFAOYSA-N 0.000 description 1
- 235000019410 glycyrrhizin Nutrition 0.000 description 1
- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 150000002430 hydrocarbons Chemical group 0.000 description 1
- 230000003463 hyperproliferative effect Effects 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000006882 induction of apoptosis Effects 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 229940069445 licorice extract Drugs 0.000 description 1
- 229940051810 licorice root extract Drugs 0.000 description 1
- 235000020725 licorice root extract Nutrition 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- CUONGYYJJVDODC-UHFFFAOYSA-N malononitrile Chemical compound N#CCC#N CUONGYYJJVDODC-UHFFFAOYSA-N 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 230000036456 mitotic arrest Effects 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000012120 mounting media Substances 0.000 description 1
- KVBGVZZKJNLNJU-UHFFFAOYSA-N naphthalene-2-sulfonic acid Chemical compound C1=CC=CC2=CC(S(=O)(=O)O)=CC=C21 KVBGVZZKJNLNJU-UHFFFAOYSA-N 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 239000012044 organic layer Substances 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 238000009116 palliative therapy Methods 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- RAIYODFGMLZUDF-UHFFFAOYSA-N piperidin-1-ium;acetate Chemical compound CC([O-])=O.C1CC[NH2+]CC1 RAIYODFGMLZUDF-UHFFFAOYSA-N 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 description 1
- INCIMLINXXICKS-UHFFFAOYSA-M pyronin Y Chemical compound [Cl-].C1=CC(=[N+](C)C)C=C2OC3=CC(N(C)C)=CC=C3C=C21 INCIMLINXXICKS-UHFFFAOYSA-M 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 235000019203 rebaudioside A Nutrition 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- XLXOKMFKGASILN-UHFFFAOYSA-N rhodamine red-X Chemical compound C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=C(S(=O)(=O)NCCCCCC(O)=O)C=C1S([O-])(=O)=O XLXOKMFKGASILN-UHFFFAOYSA-N 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 229930195734 saturated hydrocarbon Natural products 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- FHHPUSMSKHSNKW-SMOYURAASA-M sodium deoxycholate Chemical compound [Na+].C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 FHHPUSMSKHSNKW-SMOYURAASA-M 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 229960002920 sorbitol Drugs 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- IHIXIJGXTJIKRB-UHFFFAOYSA-N trisodium vanadate Chemical compound [Na+].[Na+].[Na+].[O-][V]([O-])([O-])=O IHIXIJGXTJIKRB-UHFFFAOYSA-N 0.000 description 1
- 238000001665 trituration Methods 0.000 description 1
- 230000005748 tumor development Effects 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
- C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
- C07D471/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/308—Foods, ingredients or supplements having a functional effect on health having an effect on cancer prevention
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/30—Other Organic compounds
Abstract
Description
본 발명은 신규한 LoxL2 억제제인 인돌리진 유도체 화합물 및 이를 포함하는 암, 구체적으로는 췌장암의 예방 또는 치료용 약제학적 조성물에 관한 것이다.The present invention relates to a novel LoxL2 inhibitor, an indolizine derivative compound, and a pharmaceutical composition for preventing or treating cancer including the same, specifically, pancreatic cancer.
암은 생체로부터 영양을 공급받으면서도, 생체와는 독립적으로 과잉 증식하며, 생체를 파괴하는 생체조직에서 발생한 비정상적인 조직 덩어리이다. 신체의 모든 장기는 수많은 세포로 구성되어 있는데 정상적인 세포가 과증식성의 이상 세포로 변질되면서 무질서하게 분열 증식하여 암 조직이 발생하게 된다. 암의 발병에는 유전적 소인 뿐 아니라, 환경적 요인을 비롯한 후천적 원인 또한 큰 영향을 미치며 선진국일수록 암의 발병이 증가하는 경향을 보인다.Cancer is an abnormal tissue mass generated from living tissue that, while receiving nutrients from the living body, proliferates independently of the living body and destroys the living body. All organs in the body are composed of numerous cells, and as normal cells degenerate into hyperproliferative abnormal cells, they divide and proliferate in disorder to form cancerous tissue. In addition to genetic predisposition to the onset of cancer, acquired causes, including environmental factors, also have a great influence, and the incidence of cancer tends to increase in developed countries.
췌장암은 발병률과 사망률의 거의 비슷해 5년 생존율이 8% 내외에 불과한 매우 불량한 예후를 보이는 암으로 위험성 또한 높은 것으로 나타났다. 췌장암의 병인은 유전적인 요인과 환경적 요인으로 부분적으로 설명하고 있으나 아직 그 발병 기전이 명확하지 않고, 암이 상당부분 진행되기 전까지 특별한 자각 증상이 없으며, 질병에 취약한 환자들을 선별해 낼 수 있는 구체적인 표지자가 개발되지 못하여 조기 진단이 매우 어렵다. 더욱이 췌장암 세포는 전이가 잘 되는 특성으로 인해 수술 후 높은 재발율을 보여 완치를 더욱 어렵게 하고 있다. 췌장암의 효과적인 치료를 위해 다양한 치료제 개발이 진행되어 왔지만 현재 FDA의 승인을 받은 약물은 DNA의 합성을 억제하여 암세포의 성장을 억제하는 젬시타빈(Gemcitabine)과 5-FU(5-fluorouracil)를 비롯한 보조 치료제 몇 종에 지나지 않으며 그 효과 또한 환자의 생존률을 유의미하게 개선시킬 정도의 수준이 아니다. 젬시타빈은 현재 췌장암 치료의 표준 1차 치료로서 1996년에 미국 FDA 승인이 되었으나, 국부 진행성 췌장암을 가진 126명의 환자를 대상으로 한 임상 연구에서 젬시타빈은 5-FU보다 중앙 전체생존기간, 질환 진행까지의 중앙 기간 및 임상 이익 반응면에서 우월한 것으로 나타났으나, 승인된 이후 췌장암의 표준 완화적 요법이 되었지만, 췌장암 치료에서의 개선은 거의 없었다. 때문에 췌장암은 여전히 예후가 매우 불량한 대표적인 암으로 남아있어 보다 효율적인 신규 치료제의 개발이 시급하다.Pancreatic cancer is a cancer with a very poor prognosis with a 5-year survival rate of only about 8% due to almost the same incidence and mortality rates. The pathogenesis of pancreatic cancer is partially explained by genetic and environmental factors, but its pathogenesis is not yet clear, and there are no specific symptoms until the cancer has progressed to a considerable extent. Early diagnosis is very difficult because markers are not developed. Moreover, pancreatic cancer cells have a high recurrence rate after surgery due to their ability to metastasize, making it more difficult to cure. Various therapeutic agents have been developed for the effective treatment of pancreatic cancer, but currently FDA-approved drugs, including gemcitabine and 5-fluorouracil, which inhibit the growth of cancer cells by inhibiting DNA synthesis There are only a few therapeutic agents, and their effectiveness is not at a level that significantly improves the survival rate of patients. Although gemcitabine was currently approved by the US FDA in 1996 as the standard first-line treatment for pancreatic cancer, in a clinical study of 126 patients with locally advanced pancreatic cancer, gemcitabine was superior to 5-FU for median overall survival and disease progression Although it has been shown to be superior in terms of median time to and clinical benefit response, it has become the standard palliative therapy for pancreatic cancer since it was approved, but there was little improvement in the treatment of pancreatic cancer. Therefore, pancreatic cancer still remains as a representative cancer with a very poor prognosis, so there is an urgent need to develop more efficient new therapeutic agents.
한편, LoxL2(Lysyl oxidase like protein 2)는 산화효소 활성을 갖는 단백질로서 최근 각종 암의 발생 및 전이 과정에서 중요한 역할을 한다는 것이 보고되고 있다. LoxL2는 암 전이 과정 중 EMT(epithelial to mesenchymal transition)의 진행에 관여하는 효소로서 다양한 연구에서 LoxL2의 억제가 EMT 저하 및 암 전이 차단으로 이어진다고 보고하고 있다. 따라서, LoxL2에 대한 효율적인 억제제는 암 발생 및 전이의 진행을 유의하게 저해시킬 수 있다. Meanwhile, it has been reported that LoxL2 (Lysyl oxidase like protein 2) is a protein having an oxidase activity and plays an important role in the development and metastasis of various cancers. LoxL2 is an enzyme involved in the progression of epithelial to mesenchymal transition (EMT) during cancer metastasis, and various studies have reported that inhibition of LoxL2 leads to lowering of EMT and blocking of cancer metastasis. Therefore, an efficient inhibitor of LoxL2 can significantly inhibit the progression of cancer development and metastasis.
본 명세서 전체에 걸쳐 다수의 논문 및 특허문헌이 참조되고 그 인용이 표시되어 있다. 인용된 논문 및 특허문헌의 개시 내용은 그 전체로서 본 명세서에 참조로 삽입되어 본 발명이 속하는 기술 분야의 수준 및 본 발명의 내용이 보다 명확하게 설명된다.Numerous papers and patent documents are referenced throughout this specification and citations thereof are indicated. The disclosure contents of the cited papers and patent documents are incorporated herein by reference in their entirety to more clearly describe the level of the technical field to which the present invention pertains and the content of the present invention.
본 발명자들은 종양의 발생과 전이 과정에 중요한 역할을 하는 산화 효소인 LoxL2(Lysyl oxidase like protein 2)의 활성을 특이적으로 억제함으로써 암세포의 증식, 이동 및 침윤을 저해하고 암세포의 사멸을 효율적으로 유도하는 우수한 항암 조성물을 개발하기 위하여 예의 연구 노력하였다. 그 결과, 하기 화학식 1로 표시되는 인돌리진(indolizine) 유도체가 악성 종양에서의 LoxL2 발현량 및 활성을 현저히 억제하면서 암세포의 증식과 세포분열을 억제하고, 암세포 사멸과 관련된 유전자의 발현을 크게 증가시킴으로써 다각적인 항암 활성을 발휘한다는 사실을 발견하여, 본 발명을 완성하게 되었다.The present inventors inhibit the proliferation, migration and invasion of cancer cells by specifically inhibiting the activity of Lysyl oxidase like protein 2 (LoxL2), an oxidase enzyme that plays an important role in the development and metastasis of tumors, and effectively induces apoptosis of cancer cells In order to develop an excellent anti-cancer composition, intensive research efforts were made. As a result, the indolizine derivative represented by the following Chemical Formula 1 significantly suppresses the expression and activity of LoxL2 in malignant tumors, suppresses the proliferation and cell division of cancer cells, and significantly increases the expression of genes related to cancer cell death. By discovering the fact that it exerts various anticancer activities, the present invention has been completed.
따라서 본 발명의 목적은 신규한 인돌리진 유도체 화합물 및 이를 유효성분으로 포함하는 암의 예방 또는 치료용 조성물을 제공하는 데 있다.Accordingly, an object of the present invention is to provide a novel indolizine derivative compound and a composition for preventing or treating cancer comprising the same as an active ingredient.
본 발명의 다른 목적은 LoxL2의 활성 또는 발현 억제용 조성물을 제공하는 데 있다.Another object of the present invention is to provide a composition for inhibiting the activity or expression of LoxL2.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.
본 발명의 일 양태에 따르면, 본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 약제학적으로 허용 가능한 염을 제공한다:According to one aspect of the present invention, the present invention provides a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
화학식 1
상기 화학식에서 R1은 NA1A2(A1 및 A2는 각각 독립적으로 수소 또는 C1-C3 알킬이다)이고 R2는 수소 또는 C1-C3 알킬이다. In the above formula, R 1 is NA 1 A 2 (A 1 and A 2 are each independently hydrogen or C 1 -C 3 alkyl) and R 2 is hydrogen or C 1 -C 3 alkyl.
본 발명자들은 종양의 발생과 전이 과정에 중요한 역할을 하는 산화 효소인 LoxL2(Lysyl oxidase like protein 2)의 활성을 특이적으로 억제함으로써 암세포의 증식, 이동 및 침윤을 저해하고 암세포의 사멸을 효율적으로 유도하는 우수한 항암 조성물을 개발하기 위하여 예의 연구 노력하였다. 그 결과, 상기 화학식 1로 표시되는 인돌리진(indolizine) 유도체가 악성 종양에서의 LoxL2 발현량 및 활성을 현저히 억제하면서 암세포의 증식과 세포분열을 억제하고, 암세포 사멸과 관련된 유전자의 발현을 크게 증가시킴으로써 다각적인 항암 활성을 발휘한다는 사실을 발견하였다. The present inventors inhibit the proliferation, migration and invasion of cancer cells by specifically inhibiting the activity of Lysyl oxidase like protein 2 (LoxL2), an oxidase enzyme that plays an important role in the development and metastasis of tumors, and effectively induces apoptosis of cancer cells In order to develop an excellent anti-cancer composition, intensive research efforts were made. As a result, the indolizine derivative represented by Formula 1 remarkably suppresses the expression and activity of LoxL2 in malignant tumors, suppresses the proliferation and division of cancer cells, and significantly increases the expression of genes related to cancer cell death. It was found that it exerts a multifaceted anticancer activity.
본 명세서에서 용어“알킬”은 직쇄 또는 분쇄의 포화 탄화수소기를 의미하며, 예를 들어, 메틸, 에틸, 프로필, 이소프로필 등을 포함한다. C1-C3 알킬은 탄소수 1 내지 3의 알킬 유니트를 가지는 알킬기를 의미하며, C1-C3 알킬이 치환된 경우 치환체의 탄소수는 포함되지 않은 것이다. As used herein, the term “alkyl” refers to a straight-chain or branched saturated hydrocarbon group, and includes, for example, methyl, ethyl, propyl, isopropyl, and the like. C 1 -C 3 alkyl refers to an alkyl group having an alkyl unit having 1 to 3 carbon atoms, and when C 1 -C 3 alkyl is substituted, the carbon number of the substituent is not included.
본 발명의 구체적인 구현예에 따르면, 상기 A1 및 A2는 수소이다.According to a specific embodiment of the present invention, A 1 and A 2 are hydrogen.
본 발명의 구체적인 구현예에 따르면, 상기 R2는 C1 알킬이다.According to a specific embodiment of the present invention, R 2 is C 1 alkyl.
A1 및 A2가 수소이고 R2가 C1 알킬(메틸)인 화학식 1 화합물은“6-아미노-5-(4-메톡시벤조일)인돌리진-7-카보니트릴”로서, 본 발명자들은 방대한 후보 화합물 라이브러리를 이용하여 LoxL2에 대한 억제제를 탐색한 결과, 현저히 우수한 LoxL2 억제 활성을 보이는 상기 화합물(실시예에서는“#765”로 명명함)을 발굴하였다.The compound of
본 발명의 다른 양태에 따르면, 본 발명은 전술한 본 발명의 화합물 또는 이의 약제학적으로 허용 가능한 염을 유효성분으로 포함하는 암의 예방 또는 치료용 조성물을 제공한다.According to another aspect of the present invention, the present invention provides a composition for preventing or treating cancer comprising the above-described compound of the present invention or a pharmaceutically acceptable salt thereof as an active ingredient.
본 명세서에서 용어“예방”은 질환 또는 질병을 보유하고 있다고 진단된 적은 없으나, 이러한 질환 또는 질병에 걸릴 가능성이 있는 대상체에서 질환 또는 질병의 발생을 억제하는 것을 의미한다. As used herein, the term “prevention” refers to inhibiting the occurrence of a disease or disease in a subject who has never been diagnosed as having a disease or disease, but is likely to be afflicted with the disease or disease.
본 명세서에서 용어“치료”는 (a) 질환, 질병 또는 증상의 발전의 억제; (b) 질환, 질병 또는 증상의 경감; 또는 (c) 질환, 질병 또는 증상을 제거하는 것을 의미한다. 본 발명의 조성물을 대상체에 투여하면 종양의 발생, 증식 및 전이에 핵심적인 매개체인 LoxL2의 활성을 억제함으로써, 악성 종양의 발생 및 진행을 억제하거나, 이를 제거하거나 또는 경감시키는 역할을 한다. 따라서, 본 발명의 조성물은 그 자체로 종양의 치료 조성물이 될 수도 있고, 혹은 다른 약리성분과 함께 투여되어 종양에 대한 치료 보조제로 적용될 수도 있다. 이에, 본 명세서에서 용어“치료”또는“치료제”는“치료 보조”또는“치료 보조제”의 의미를 포함한다. As used herein, the term “treatment” refers to (a) inhibiting the development of a disease, disorder or condition; (b) alleviation of the disease, condition or condition; or (c) eliminating the disease, condition or symptom. When the composition of the present invention is administered to a subject, it inhibits the activity of LoxL2, which is a key mediator for tumor development, proliferation, and metastasis, thereby inhibiting, eliminating, or alleviating the development and progression of malignant tumors. Accordingly, the composition of the present invention may be a therapeutic composition for a tumor itself, or may be administered with other pharmacological components and applied as a therapeutic adjuvant for a tumor. Accordingly, as used herein, the term “treatment” or “therapeutic agent” includes the meaning of “therapeutic adjuvant” or “therapeutic adjuvant”.
본 명세서에서 용어“투여”또는“투여하다”는 본 발명의 조성물의 치료적 유효량을 대상체에 직접적으로 투여함으로써 대상체의 체내에서 동일한 양이 형성되도록 하는 것을 말한다.As used herein, the term “administration” or “administering” refers to direct administration of a therapeutically effective amount of the composition of the present invention to a subject so that the same amount is formed in the subject's body.
본 발명에서 용어“치료적 유효량”은 본 발명의 약제학적 조성물을 투여하고자 하는 개체에게 조성물 내의 약리성분이 치료적 또는 예방적 효과를 제공하기에 충분한 정도로 함유된 조성물의 함량을 의미하며, 이에“예방적 유효량”을 포함하는 의미이다. In the present invention, the term “therapeutically effective amount” refers to the content of the composition in which the pharmacological component in the composition is sufficient to provide a therapeutic or prophylactic effect to an individual to whom the pharmaceutical composition of the present invention is to be administered. prophylactically effective amount”.
본 명세서에서 용어“대상체”는 제한없이 인간, 마우스, 래트, 기니아 피그, 개, 고양이, 말, 소, 돼지, 원숭이, 침팬지, 비비 또는 붉은털 원숭이를 포함한다. 구체적으로는, 본 발명의 대상체는 인간이다. As used herein, the term “subject” includes, without limitation, humans, mice, rats, guinea pigs, dogs, cats, horses, cattle, pigs, monkeys, chimpanzees, baboons or rhesus monkeys. Specifically, the subject of the present invention is a human.
본 명세서에서 용어 "약제학적으로 허용되는 염"은 약학적으로 허용되는 무기산, 유기산, 또는 염기로부터 유도된 염을 포함한다. 적합한 산의 예로는 염산, 브롬산, 황산, 질산, 과염소산, 푸마르산, 말레산, 인산, 글리콜산, 락트산, 살리실산, 숙신산, 톨루엔-p-설폰산, 타르타르산, 아세트산, 트리플루로초산, 시트르산, 메탄설폰산, 포름산, 벤조산, 말론산, 나프탈렌-2-설폰산, 벤젠설폰산 등을 들 수 있다. 적합한 염기로부터 유도된 염은 나트륨 등의 알칼리 금속, 마그네슘 등의 알칼리 토금속, 및 암모늄 등을 포함할 수 있다.As used herein, the term "pharmaceutically acceptable salt" includes salts derived from pharmaceutically acceptable inorganic acids, organic acids, or bases. Examples of suitable acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, trifluoroacetic acid, citric acid, methane sulfonic acid, formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals such as sodium, alkaline earth metals such as magnesium, and ammonium and the like.
본 발명의 구체적인 구현예에 따르면, 본 발명의 조성물로 예방 또는 치료될 수 있는 암은 췌장암, 유방암, 대장암 및 위암으로 구성된 군으로부터 선택된다. 보다 구체적으로는, 상기 암은 췌장암이다.According to a specific embodiment of the present invention, the cancer that can be prevented or treated by the composition of the present invention is selected from the group consisting of pancreatic cancer, breast cancer, colorectal cancer and gastric cancer. More specifically, the cancer is pancreatic cancer.
본 발명의 구체적인 구현예에 따르면, 본 발명의 조성물은 JNK(c-Jun N-terminal kinase)를 활성화시키고, 미세관의 중합을 저해함으로써 암세포의 증식을 억제한다. 따라서, 본 발명의 조성물은 LoxL2의 억제 및 미세관 탈중합화(microtubule depolymerization)라는 이중의 항암 메카니즘을 통해 다각적으로 암세포를 사멸시키며, 이에 LoxL2 양성 암 뿐 아니라 LoxL2 음성 암에 대해서도 유의한 치료 효과를 발휘한다. According to a specific embodiment of the present invention, the composition of the present invention inhibits the proliferation of cancer cells by activating c-Jun N-terminal kinase (JNK) and inhibiting the polymerization of microtubules. Therefore, the composition of the present invention kills cancer cells in various ways through the dual anticancer mechanism of LoxL2 inhibition and microtubule depolymerization, and thus has a significant therapeutic effect on LoxL2-positive cancer as well as LoxL2-negative cancer. perform
본 명세서에서 용어“LoxL2 양성 암”은 세포 또는 조직에서 LoxL2 효소의 발현이 검출되는 고형암 또는 혈액암을 의미한다.As used herein, the term “LoxL2-positive cancer” refers to a solid cancer or blood cancer in which LoxL2 enzyme expression is detected in cells or tissues.
본 발명의 조성물이 약제학적 조성물로 제조되는 경우, 본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체를 포함한다.When the composition of the present invention is prepared as a pharmaceutical composition, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier.
본 발명의 약제학적 조성물에 포함되는 약제학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. 적합한 약제학적으로 허용되는 담체 및 제제는 Remington's Pharmaceutical Sciences (19th ed., 1995)에 상세히 기재되어 있다.Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. it's not going to be The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).
본 발명의 약제학적 조성물은 경구 또는 비경구 투여할 수 있으며, 구체적으로는 경구, 정맥, 피하 또는 복강 투여될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally, and specifically may be administered orally, intravenously, subcutaneously or intraperitoneally.
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 바람직한 투여량은 성인 기준으로 0.001-1000 ㎎/kg 범위 내이다.A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity of the patient. can be A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001-1000 mg/kg for adults.
본 발명의 약제학적 조성물은 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액, 시럽제 또는 유화액 형태이거나 엑스제, 산제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.
본 발명의 또 다른 양태에 따르면, 본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 식품학적으로 허용가능한 염을 유효성분으로 포함하는 암의 예방 또는 개선용 기능성 식품 조성물을 제공한다:According to another aspect of the present invention, the present invention provides a functional food composition for the prevention or improvement of cancer comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
화학식 1
본 발명에서 이용되는 화학식 1 화합물 및 이를 이용하여 예방 또는 개선될 수 있는 암에 대해서는 이미 상술하였으므로, 과도한 중복을 피하기 위해 그 기재를 생략한다.Since the compound of
본 명세서에서 용어 "식품학적으로 허용되는 염"은, 양이온과 음이온이 정전기적 인력에 의해 결합하는 염 중에서도 식품 조성물에 사용될 수 있는 형태의 염을 의미하며, 그 구체적인 예는 상술한 "약제학적으로 허용되는 염"의 예를 포함한다.As used herein, the term "food pharmaceutically acceptable salt" refers to a salt in a form that can be used in a food composition among salts in which cations and anions are combined by electrostatic attraction, and specific examples thereof include the above-mentioned "pharmaceutically acceptable salts". acceptable salts".
본 발명의 또 다른 양태에 따르면, 본 발명은 하기 화학식 1로 표시되는 화합물 또는 이의 식품학적으로 허용가능한 염을 유효성분으로 포함하는 LoxL2(Lysyl oxidase like 2)의 활성 억제용 기능성 식품 조성물을 제공한다:According to another aspect of the present invention, the present invention provides a functional food composition for inhibiting the activity of LoxL2 (Lysyl oxidase like 2) comprising a compound represented by the following
화학식 1
본 발명에서 이용되는 화학식 1 화합물에 대해서는 이미 상술하였으므로, 과도한 중복을 피하기 위해 그 기재를 생략한다.Since the compound of
본 명세서에서 용어“활성의 억제”는 목적 단백질, 즉 LoxL2 효소의 활성 또는 발현의 저하를 야기하는 것을 의미하며, 이에 의해 LoxL2의 활성 또는 발현이 탐지 불가능해지거나 무의미한 수준으로 존재하게 되는 경우 뿐 아니라, LoxL2의 산화 효소로서의 생물학적 기능이 유의하게 저해되어 종양의 발생, 증식 및 전이와 이로 인한 증상의 악화가 측정 가능한 수준으로 개선될 정도로 LoxL2 효소의 발현량 또는 LoxL2 효소의 생체 내 고유한 기능이 감소하는 것을 의미한다. 구체적으로는 대조군에 비하여 활성 또는 발현량이 20% 이상 감소한 상태, 보다 구체적으로는 40% 이상 감소한 상태, 더욱 구체적으로는 60% 이상 감소한 상태를 의미할 수 있다.As used herein, the term “inhibition of activity” means to cause a decrease in the activity or expression of a target protein, that is, the LoxL2 enzyme, whereby the activity or expression of LoxL2 becomes undetectable or exists at an insignificant level, as well as , LoxL2 enzyme expression level or the intrinsic function of LoxL2 enzyme in vivo is reduced to such an extent that the biological function of LoxL2 as an oxidase is significantly inhibited, and the occurrence, proliferation, and metastasis of tumors and worsening of symptoms thereof are improved to a measurable level. means to do Specifically, it may refer to a state in which the activity or expression level is reduced by 20% or more, more specifically, a state in which the activity or expression is reduced by more than 40%, more specifically, a state in which the activity or expression level is reduced by more than 60% compared to the control.
본 발명의 조성물이 식품 조성물로 제조되는 경우, 유효성분으로서 본 발명의 화합물 뿐 만 아니라, 식품 제조 시에 통상적으로 첨가되는 탄수화물, 조미제 및 향미제를 포함할 수 있다. 탄수화물의 예는 포도당, 과당 등의 단당류; 말토스, 수크로스 등의 이당류 및 덱스트린, 사이클로덱스트 린 등과 같은 다당류 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜을 포함하나 이에 제한되는 것은 아니다. 향미제로서 천연 향미제[타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스 파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제로 제조되는 경우에는 본 발명의 유효성분인 #765 화합물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 두충 추출액, 대추 추출액, 감초 추출액 등을 추가로 포함시킬 수 있다.When the composition of the present invention is prepared as a food composition, it may include not only the compound of the present invention as an active ingredient, but also carbohydrates, seasonings and flavoring agents that are commonly added during food production. Examples of carbohydrates include monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol, but are not limited thereto. As the flavoring agent, natural flavoring agents (taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, licorice root extract, jujube extract, licorice extract, etc. are added in addition to the #765 compound, which is the active ingredient of the present invention. can be included as
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 신규한 인돌리진 유도체 화합물 및 이를 유효성분으로 포함하는 암의 예방 또는 치료용 조성물을 제공한다.(a) The present invention provides a novel indolizine derivative compound and a composition for preventing or treating cancer comprising the same as an active ingredient.
(b) 본 발명의 인돌리진 유도체는 종양의 발생과 전이 과정에 핵심적인 매개체로 알려진 LoxL2 효소의 활성을 현저히 억제함으로써 췌장암을 비롯한 다양한 암에 대한 효율적인 항암제 조성물로 유용하게 이용될 수 있다.(b) The indolizine derivative of the present invention can be usefully used as an effective anticancer composition for various cancers including pancreatic cancer by remarkably inhibiting the activity of LoxL2 enzyme, which is known as a key mediator in the development and metastasis of tumors.
(c) 본 발명의 인돌리진 유도체는 또한 종양세포의 증식 과정에서 미세관(microtubule)의 중합화를 저해하여 세포주기를 유사분열(mitosis)에서 정지시킴으로써 췌장암을 비롯한 증식이 빠른 종양에 대해 현저한 암세포 사멸 효과를 나타낸다.(c) The indolizine derivative of the present invention also inhibits the polymerization of microtubules in the process of tumor cell proliferation, thereby stopping the cell cycle from mitosis, thereby making it a remarkable cancer cell for fast-growing tumors including pancreatic cancer. exhibits a killing effect.
도 1은 은 췌장암 세포의 증식에 대한 #765의 효과를 나타낸 것이다.
도 2는 췌장암 세포의 콜로니(colony) 형성에 대한 #765의 효과를 나타낸 것이다.
도 3은 유세포분석법(flow cytometry)을 이용하여 #765를 처리한 췌장암 세포의 세포 주기를 분석한 것이다.
도 4는 웨스턴 블롯(western blot)과 면역형광염색법(immunofluorescence assay)을 이용하여 #765를 처리한 췌장암 세포에서 세포 주기와 관련된 단백질의 발현을 분석한 것이다.
도 5는 미세관 역동학(microtubule dynamics)에 대한 #765의 효과를 나타낸 것이다.
도 6은 유세포분석법(flow cytometry)을 이용하여 췌장암 세포와 정상 췌장 표피 세포에서의 아폽토시스에 대한 #765의 효과를 나타낸 것이다.
도 7은 웨스턴 블롯를 이용하여 #765를 처리한 췌장암 세포에서 세포사멸 관련 단백질의 발현 변화를 분석한 결과를 나타낸다.
도 8은 Mia PaCa-2 세포에서 #765에 의해 유도되는 세포사멸에 있어서의 JNK의 영향을 보여주는 그림이다.1 shows the effect of #765 on the proliferation of pancreatic cancer cells.
Figure 2 shows the effect of #765 on colony formation of pancreatic cancer cells.
3 is a cell cycle analysis of pancreatic cancer cells treated with #765 using flow cytometry.
4 is an analysis of the expression of cell cycle-related proteins in pancreatic cancer cells treated with #765 using western blot and immunofluorescence assay.
Figure 5 shows the effect of #765 on microtubule dynamics.
6 shows the effect of #765 on apoptosis in pancreatic cancer cells and normal pancreatic epithelial cells using flow cytometry.
7 shows the results of analyzing the expression change of apoptosis-related proteins in pancreatic cancer cells treated with #765 using Western blot.
8 is a diagram showing the effect of JNK on apoptosis induced by #765 in Mia PaCa-2 cells.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 요지에 따라 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .
실시예Example
실험방법Experimental method
화합물 #765의 합성Synthesis of
1-(2-(4-메톡시페닐)-2-옥소에틸)-1H-피롤-2-카브알데하이드(전구체)의 합성Synthesis of 1-(2-(4-methoxyphenyl)-2-oxoethyl)-1H-pyrrole-2-carbaldehyde (precursor)
피롤-2-카복스알데하이드(300 mg, 3.155 mmol)를 CH3CN(10 ㎖)에 녹이고, K2CO3(654 mg, 1.5 당량) 및 2-브로모-4′-메톡시아세토페논(867.1 mg, 1.2 당량)을 넣은 뒤 상온에서 저어주었다. 24시간 동안 저어준 후, 반응 혼합액을 감압 농축하고, 에틸아세테이트(18 ㎖)로 희석한 다음 H2O(18 ㎖)로 세척하였다. 에틸아세테이트(18 ㎖)를 사용하여 수층을 다시 한 번 추출하였다. MgSO4로 유기물 층을 건조시키고, 감압 농축하였다. 얻은 잔사물을 혼합 용매(헥산 : 에틸아세테이트 : 디클로로메탄 = 30 : 1 : 2 또는 10 : 1 : 2)로 마쇄(trituration)하여 정제함으로써 목적 화합물(629.3 mg, 82%)을 수득하였다.Dissolve pyrrole-2-carboxaldehyde (300 mg, 3.155 mmol) in CH 3 CN (10 mL), K 2 CO 3 (654 mg, 1.5 equiv) and 2-bromo-4′-methoxyacetophenone ( 867.1 mg, 1.2 equivalents) was added and stirred at room temperature. After stirring for 24 hours, the reaction mixture was concentrated under reduced pressure, diluted with ethyl acetate (18 mL), and washed with H 2 O (18 mL). The aqueous layer was extracted once again using ethyl acetate (18 mL). The organic layer was dried over MgSO 4 and concentrated under reduced pressure. The obtained residue was purified by trituration with a mixed solvent (hexane : ethyl acetate : dichloromethane = 30 : 1 : 2 or 10 : 1 : 2) to obtain the target compound (629.3 mg, 82%).
6-아미노-5-(4-메톡시벤조일)인돌리진-7-카보니트릴(#765)의 합성Synthesis of 6-amino-5-(4-methoxybenzoyl)indolizine-7-carbonitrile (#765)
화합물 e(56 mg, 0.23 mmol)를 EtOH 2 ㎖에 녹이고, 피페리디늄 아세테이트(17 mg, 0.5 당량)와 말로노나이트릴(화합물 C; 22.8 mg, 1.5 당량)을 넣은 뒤, 120℃에서 24시간 동안 반응시켰다. 반응 종결 후 반응물을 감압농축 하여 얻은 잔사물을 실리카 겔 컬럼 크로마토그래피(헥산 : 에틸 아세테이트 : 디클로로메테인 = 10 : 1 : 2)로 정제하여 목적 화합물을 수득하였다.Compound e (56 mg, 0.23 mmol) was dissolved in 2 mL of EtOH, piperidinium acetate (17 mg, 0.5 equiv) and malononitrile (compound C; 22.8 mg, 1.5 equiv) were added thereto, followed by 24 at 120°C. reacted for hours. After completion of the reaction, the residue obtained by concentrating the reaction product under reduced pressure was purified by silica gel column chromatography (hexane : ethyl acetate : dichloromethane = 10 : 1 : 2) to obtain the target compound.
화학식 #765
오렌지색 고형물, mp: 131.4~132.2℃ (64.3 mg, 96%); 1 H NMR (400 MHz, CDCl3) δ 7.83 (s, 1H), 7.54 (d, J = 8.4 Hz, 2H), 7.01 (s, 1H), 6.89 (d, J = 8.8 Hz, 2H), 6.72 (d, J = 4.0 Hz, 1H), 6.53 (dd, J = 2.4, 3.8 Hz,1H), 5.83 (s, 2H), 3.87 (s, 3H); 13 C NMR (100 MHz, CDCl3) δ 188.5, 163.5, 139.3, 131.0, 130.8, 130.1, 128.4, 122.6, 116.7, 114.5, 114.4, 113.5, 108.2, 93.8, 55.6; HRMS (ESI-QTOF) calcd for C17H13N3O 2292.1081 ([M+H]+), found 292.1076. orange solid, mp: 131.4-132.2° C. (64.3 mg, 96%); 1 H NMR (400 MHz, CDCl 3 ) δ 7.83 (s, 1H), 7.54 (d, J = 8.4 Hz, 2H), 7.01 (s, 1H), 6.89 (d, J = 8.8 Hz, 2H), 6.72 (d, J = 4.0 Hz, 1H), 6.53 (dd, J = 2.4, 3.8 Hz, 1H), 5.83 (s, 2H), 3.87 (s, 3H); 13 C NMR (100 MHz, CDCl 3 ) δ 188.5, 163.5, 139.3, 131.0, 130.8, 130.1, 128.4, 122.6, 116.7, 114.5, 114.4, 113.5, 108.2, 93.8, 55.6; HRMS (ESI-QTOF) calcd for C 17 H 13 N 3 O 2292.1081 ([M+H] + ), found 292.1076.
세포배양 cell culture
인간 췌장암 세포주(Mia PaCa-2, PANC-1, BxPC-3, AsPC-1)는 ATCC(American Type Culture Collection; Manassas, VA, USA)에서 구입하였고, 정상 인간 췌장 상피세포(HPDE)는 방승민 교수(연세대학교 의과대학)로부터 제공받았다. Mia PaCa-2는 DMEM(Dulbecco’s modified Eagle’s medium; Hyclone, Logan, UT, USA)에 2.5% 말혈청(Gibco, New York, NY, USA), 불활성화시킨 10% 우태아혈청(FBS; Hyclone), 1% 페니실린/스트렙토마이신(p/s; Hyclone)을 첨가하여 배양하였다. PANC-1은 DMEM에 10% FBS, 1% p/s를 첨가하여 배양하였다. BxPC-3와 AsPC-1은 RPMI (Hyclone)에 10% FBS, 1% p/s를 첨가하여 배양하였다. HPDE는 각질형성세포 없는 배지에 인간 재조합 표피성장인자(Gibco), 소 뇌하수체 추출물(Gibco), 1% p/s를 첨가하여 배양하였다. 모든 세포는 37℃ 및 5% CO2 가습 조건에서 유지되었다. Human pancreatic cancer cell lines (Mia PaCa-2, PANC-1, BxPC-3, AsPC-1) were purchased from ATCC (American Type Culture Collection; Manassas, VA, USA), and normal human pancreatic epithelial cells (HPDE) were obtained from Bang Seung-min. It was provided by a professor (Yonsei University College of Medicine). Mia PaCa-2 is DMEM (Dulbecco's modified Eagle's medium; Hyclone, Logan, UT, USA) in 2.5% horse serum (Gibco, New York, NY, USA), inactivated 10% fetal bovine serum (FBS; Hyclone), 1% penicillin/streptomycin (p/s; Hyclone) was added and cultured. PANC-1 was cultured by adding 10% FBS and 1% p/s to DMEM. BxPC-3 and AsPC-1 were cultured by adding 10% FBS and 1% p/s to RPMI (Hyclone). HPDE was cultured by adding human recombinant epidermal growth factor (Gibco), bovine pituitary extract (Gibco), and 1% p/s to a keratinocyte-free medium. All cells were maintained at 37° C. and 5% CO 2 humidified conditions.
세포증식 어세이cell proliferation assay
약제가 세포 증식에 미치는 영향을 세포계수 킷-8(CCK-8; Dojindo Molecular Technologies Inc., Kumamoto, Japan)을 사용하여 평가하였다. 96-웰 플레이트에 웰당 2.5 x 103개의 세포를 넣고 37℃에서 24시간 동안 배양하였다. 그 후, 본 발명의 화합물을 농도 별로 3중(triplicate)으로 처리하였으며, 양성 대조군으로 췌장암 1차 항암제인 젬시타빈(gemcitabine)을 함께 사용하였다. 약제의 용매로 사용되는 DMSO의 최종 농도는 1% 이하이다. 약물 처리 72 시간 후, CCK-8 용액(2-[2-메톡시-4-니트로페닐]-3-[4-니트로페닐]-5-[2,4-디설포닐]-2H-테트라졸륨)을 웰에 있는 배지의 1/10의 양을 처리하여 색이 나타날 때까지 배양하였다. 흡광도는 450nm에서 Epoch 마이크로플레이트 분광광도계(BioTek Instruments Inc., Winooski, VT, USA)을 이용하여 측정하였다.The effect of the drug on cell proliferation was evaluated using a cell counting kit-8 (CCK-8; Dojindo Molecular Technologies Inc., Kumamoto, Japan). 2.5 x 10 3 cells per well were placed in a 96-well plate and incubated at 37° C. for 24 hours. Thereafter, the compound of the present invention was treated in triplicate for each concentration, and gemcitabine, a primary anticancer agent for pancreatic cancer, was used together as a positive control. The final concentration of DMSO used as a solvent for pharmaceuticals is 1% or less. 72 hours after drug treatment, CCK-8 solution (2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfonyl]-2H-tetrazolium) was treated with 1/10 of the medium in the well and cultured until color appeared. Absorbance was measured at 450 nm using an Epoch microplate spectrophotometer (BioTek Instruments Inc., Winooski, VT, USA).
콜로니 형성 분석Colony formation assay
암세포의 콜로니 형성에 대한 약제의 영향을 확인하기 위해 콜로니 형성 분석이 실행되었다. 6-웰 플레이트에 웰당 500 개의 세포를 도말하고 37℃에서 24시간 동안 배양하였다. 이후 약물을 48시간 동안 처리하고, 새로운 배지로 교체한 뒤 9일 동안 배양하여 콜로니가 형성될 수 있도록 하였다. 그 후 100% 에탄올로 고정하고 0.5% 크리스탈 바이올렛을 이용하여 염색하였다.A colony formation assay was performed to confirm the effect of the drug on the colony formation of cancer cells. 500 cells per well were plated in 6-well plates and incubated at 37° C. for 24 hours. Thereafter, the drug was treated for 48 hours, replaced with a fresh medium, and cultured for 9 days to allow colonies to be formed. Thereafter, it was fixed with 100% ethanol and stained with 0.5% crystal violet.
유세포 분석법을 이용한 세포주기 분석 Cell cycle analysis using flow cytometry
암세포를 6-웰 플레이트에 웰당 2.0 - 3.0 x 105 개 세포를 도말하고 37℃에서 24시간 동안 배양하였다. 이후 약물을 24시간 동안 처리한 후, 세포를 수확하여 차가운 FACS 용액(1% FBS를 첨가한 PBS)으로 2번 씻고 70% 에탄올로 -20℃에서 하루 동안 고정시켰다. 이후 차가운 FACS 용액으로 2회 세척하고 실온에서 어둡게 하여 RNase와 함께 7-AAD로 30분 동안 염색하였다. 이후 유세포 분석기(FACS Celesta, BD Biosciences, San Jose, CA, USA)를 통해 세포당 DNA 양을 측정하여 세포주기를 분석하였다.Cancer cells were plated at 2.0 - 3.0
면역형광염색법Immunofluorescence staining
4-웰 챔버 슬라이드에 웰당 6.25 x 104개의 세포를 도말하고 37℃에서 24시간 동안 배양하였다. 이후 약물을 24시간(p-HH3 염색) 혹은 18시간(tubulin 염색) 동안 처리하였고, 양성대조군으로 mitotic arrest를 유발하는 항암제인 파클리탁셀과 노코다졸을 사용하였다. 이후 세포를 PBS로 세척하고, 메탄올을 사용하여 4℃에서 20분간 고정하였다. 그 후 0.1% PBS-T (PBS에 Triton X-100 첨가)로 세척하고, 실온에서 1시간 동안 0.1% PBS-T로 만든 20% 정상 고트 혈청으로 블로킹을 진행하였다. 블로킹이 끝난 후, 항-phospho-Histone H3 항체(Ser10; #JBC1377950; Upstate, Billerica, MA, USA) 또는 항-α-튜불린 항체(#sc-32293; Santa Cruz Biotechnology, Dallas, TX, USA)를 20% 정상 고트 혈청에 희석하여 4℃에서 하루 동안 세포를 염색하였다. 그런 다음, 0.1% PBS-T로 세포를 3회 세척하고, Rhodamine Red-X-접합 항-래빗 IgG 또는 Alexa Flour 488-접합 항-마우스 IgG를 사용하여 실온에서 1시간 동안 세포를 염색하였다. 이후, 0.1% PBS-T로 세포를 3회 세척하고, VECTASHIELD Antifade mounting Medium with DAPI(Vector Laboratories Inc., Burlingame, CA, USA)를 사용하여 DAPI 염색과 마운팅을 진행하였다. 이와 같은 과정을 거친 슬라이드를 공초점 현미경(Olympus FlouView FV1000; Olympus, Waltham, MA, USA)으로 촬영하였다.6.25 x 10 4 cells per well were plated on 4-well chamber slides and incubated at 37°C for 24 hours. Then, the drug was treated for 24 hours (p-HH3 staining) or 18 hours (tubulin staining), and paclitaxel and nocodazole, which are anticancer agents that induce mitotic arrest, were used as positive controls. Thereafter, the cells were washed with PBS and fixed at 4° C. for 20 minutes using methanol. Then, it was washed with 0.1% PBS-T (Triton X-100 added to PBS), and blocking was performed with 20% normal goat serum made of 0.1% PBS-T at room temperature for 1 hour. After blocking, anti-phospho-Histone H3 antibody (Ser10; #JBC1377950; Upstate, Billerica, MA, USA) or anti-α-tubulin antibody (#sc-32293; Santa Cruz Biotechnology, Dallas, TX, USA) was diluted in 20% normal goat serum and stained for one day at 4°C. Then, cells were washed 3 times with 0.1% PBS-T and cells were stained with Rhodamine Red-X-conjugated anti-rabbit IgG or Alexa Flour 488-conjugated anti-mouse IgG for 1 hour at room temperature. Thereafter, the cells were washed three times with 0.1% PBS-T, and DAPI staining and mounting were performed using VECTASHIELD Antifade mounting Medium with DAPI (Vector Laboratories Inc., Burlingame, CA, USA). The slides subjected to this process were photographed with a confocal microscope (Olympus FlouView FV1000; Olympus, Waltham, MA, USA).
튜불린 중합 분석법 (In vitro tubulin polymerization assay)In vitro tubulin polymerization assay
튜불린 중합화 어세이 킷(BK006P; Cytoskeleton Inc., Denver, CO, USA)를 이용하여 미세관 결합을 평가하였다. 각 조건에서 정제된 돼지의 튜불린을 4℃ 상태를 유지하며 튜불린 중합화 완충액(80 mM PIPES pH6.9, 2mM MgCl2, 0.5mM EGTA, 1mM GTP, 및 10.2% 글리세롤)에 넣고, 즉시 미리 데워진 96-웰 플레이트로 옮긴 후, 1분 마다 340nm에서의 흡광도를 60분 동안 37℃ 플레이트 리더(Flexstation3; Molecular Devices Inc., Sunnyvale, CA, USA)를 이용하여 측정하였다.Microtubule binding was assessed using a tubulin polymerization assay kit (BK006P; Cytoskeleton Inc., Denver, CO, USA). In each condition, purified pig tubulin was placed in a tubulin polymerization buffer (80 mM PIPES pH6.9, 2 mM MgCl 2 , 0.5 mM EGTA, 1 mM GTP, and 10.2% glycerol) while maintaining a state of 4° C., and immediately in advance. After transfer to a warmed 96-well plate, absorbance at 340 nm every 1 min was measured using a 37° C. plate reader (Flexstation3; Molecular Devices Inc., Sunnyvale, CA, USA) for 60 min.
유세포분석법을 이용한 세포사멸 분석Apoptosis analysis using flow cytometry
6-웰 플레이트에 웰당 1.0 - 2.0 x 105 개의 세포를 도말하고 37℃에서 24시간 동안 배양하였다. 약물을 48시간 동안 처리한 후 세포를 수확하여 차가운 1 x AnnexinⅤ 결합 완충액으로 2회 세척하였다. 그 후 같은 용액을 사용하여 APC-conjugated AnnexinⅤ (BD Pharmingen, Sparks, MD, USA)와 7-AAD (BD Pharmingen)으로 실온에서 빛을 차단하고 20분 간 세포를 염색하였다. 이후 유세포 분석기(FACSCelesta)를 통해 세포사멸을 분석하였다1.0 - 2.0
웨스턴 블롯western blot
세포를 37℃에서 24시간 동안 배양한 후, 약물을 24시간 혹은 48시간 동안 처리하였다. 이후 세포를 수확하여 차가운 PBS로 2회 세척한 뒤, 1 x 프로테아제 억제제 칵테일과 0.5% 오소바나데이트 나트륨을 첨가한 RIPA 완충액(50 mM Tris-Cl [pH 7.5], 150 mM NaCl, 1% NP-40, 0.5% 디옥시콜산 나트륨, 0.1% 황산도데실나트륨[SDS])을 이용하여 얼음에서 30분간 용해하였다. 용해된 세포를 4℃에서 12,000 rpm에 20분간 원심분리한 후, 상층액을 수득하였다. 단백질의 농도는 BCA 어세이 킷(Thermo Fisher, Waltham, MA, USA)을 사용하여 측정하였다. 추출된 단백질은 2-머캅토에탄올을 첨가한 4 x SDS 샘플 완충액(2% SDS, 50 mM Tris-HCl [pH 6.8], 0.4 mM EDTA(pH 8.5), 10% 글리세롤, 0.002% 피로닌 Y)를 이용하여 농도를 맞춰주었다. 이후 동일한 양의 단백질을 SDS 폴리아크릴아마이드 겔을 사용하여 전기영동을 하였고, 반건조 트랜스퍼를 이용하여 0.45μm 니트로셀룰로오스 막(Invitrogen, Carlsbad, CA, USA)에 옮겼다. 폰소 S 염색 용액(Sigma Aldrich, St. Louis, MO, USA)을 이용하여 트랜스퍼된 단백질을 확인하였다. 이후 TBS-T[0.1% Tween-20를 함유한 TBS(Tris-buffered saline)]를 이용하여 만든 5% 탈지유로 1시간 동안 블로킹하였다. 1차 항체는 3% BSA에 희석하여 4℃에서 하루 동안 교반하였다. 다음날 TBS-T로 3번 씻어준 후, HRP(horseradish peroxidase)가 접합된 2차 항체를 5% 탈지유에 희석하여 실온에서 1시간 30분 동안 교반하였다. 이후 TBS-T를 이용하여 6번 씻어준 다음, X-Ray 필름(AGFA, Mortsel, Belgium)과 WesternBright ECL Chemiluminescent HRP Substrate (Advansta, Menlo Park, CA, USA)를 사용하여 타겟 단백질의 발현을 확인하였다. After incubating the cells at 37° C. for 24 hours, the drug was treated for 24 hours or 48 hours. Then, the cells were harvested, washed twice with cold PBS, and RIPA buffer (50 mM Tris-Cl [pH 7.5], 150 mM NaCl, 1% NP-) supplemented with 1 x protease inhibitor cocktail and 0.5
사용한 1차 항체는 다음과 같다: 항-β-actin (AC-15; Sigma Aldrich), 항-phospho-Cdc2 (Tyr15; #4539S; Cell Signaling Technology, Danvers, MA, USA), 항-Cdc2 (#28439S; Cell Signaling Technology), 항-cyclin B1 (#4135S; Cell Signaling Technology), 항-phospho-Histone H3 (Ser10; #JBC1377950 ; Upstate), 항-PARP (#9542S; Cell Signaling Technology), 항-caspase3 (#9662S; Cell Signaling Technology), 항-Bcl-2 (#556354; BD Pharmingen), 항-Bim (#2933S; Cell Signaling Technology), 항-phospho-JNK (Thr183/Tyr185; #9251S; Cell Signaling Technology), 항-JNK (#9252; Cell Signaling Technology).The primary antibodies used were: anti-β-actin (AC-15; Sigma Aldrich), anti-phospho-Cdc2 (Tyr15; #4539S; Cell Signaling Technology, Danvers, MA, USA), anti-Cdc2 (# 28439S; Cell Signaling Technology), anti-cyclin B1 (#4135S; Cell Signaling Technology), anti-phospho-Histone H3 (Ser10; #JBC1377950; Upstate), anti-PARP (#9542S; Cell Signaling Technology), anti-caspase3 (#9662S; Cell Signaling Technology), anti-Bcl-2 (#556354; BD Pharmingen), anti-Bim (#2933S; Cell Signaling Technology), anti-phospho-JNK (Thr183/Tyr185; #9251S; Cell Signaling Technology) ), anti-JNK (#9252; Cell Signaling Technology).
실험결과Experiment result
인간 췌장암 세포주의 증식 억제 효과 Inhibitory effect on proliferation of human pancreatic cancer cell line
인간 췌장암 세포주 Mia PaCa-2, PANC-1, BxPC-3, AsPC-1의 세포 증식에 대한 #765의 영향을 CCK-8 평가법을 통해 확인하였으며, 그 결과를 도 1 및 표 1에 나타내었다. 도 1에 나타난 바와 같이, #765는 농도 의존적으로 4가지 인간 췌장암 세포의 증식을 억제하였다. 췌장암의 1차 약제(first line drug)로 사용되는 젬시타빈(gemcitabine)과 비교하였을 때, #765는 췌장암 세포에 대해서 nM 수준에서 세포증식 억제 효능이 매우 강하게 나타났다. #765의 50% 저해 농도를 표 1에 나타내었다. 도 2에 나타난 바와 같이, #765는 Mia PaCa-2와 PANC-1의 콜로니 형성을 효과적으로 억제하였다. The effect of #765 on the cell proliferation of the human pancreatic cancer cell lines Mia PaCa-2, PANC-1, BxPC-3, and AsPC-1 was confirmed through the CCK-8 assay, and the results are shown in FIG. 1 and Table 1. As shown in FIG. 1 , #765 inhibited the proliferation of 4 types of human pancreatic cancer cells in a concentration-dependent manner. Compared with gemcitabine used as a first line drug for pancreatic cancer, #765 showed a very strong cell proliferation inhibitory effect on pancreatic cancer cells at the nM level. The 50% inhibitory concentration of #765 is shown in Table 1. As shown in FIG. 2 , #765 effectively inhibited colony formation of Mia PaCa-2 and PANC-1.
인간 췌장암 세포의 세포 주기 관련 단백질 조절 및 유사분열을 중단 효과Cell cycle-related protein regulation and mitosis-interrupting effect in human pancreatic cancer cells
#765에 의한 세포증식 억제효과가 세포 주기 진행의 억제로 인해 발생하는 것인지를 조사하기 위해, #765의 처리 후 췌장암 세포주의 세포 주기 진행을 유세포 분석을 통해 확인하였다. 그 결과, 도 3에 나타난 바와 같이 #765를 세포에 처리하자 Mia PaCa-2와 PANC-1 모두에서 G2/M 단계에 있는 세포가 축적되고 G1 및 S 단계의 세포는 감소함을 관찰하였다. 또한, #765를 고농도로 처리하자 subG1 단계에 있는 세포가 증가하는 것으로 보아, #765가 세포의 죽음을 유도한다는 것을 알 수 있다. 세포 주기를 조절하는 단백질의 발현을 웨스턴 블롯을 이용하여 확인한 결과, 도 4A에 나타난 바와 같이 #765를 처리하자 Mia PaCa-2 와 PANC-1에서 G2 단계에서 M 단계로 진행하는 데에 관여하는 것으로 알려진 Tyr15번 잔기-인산화된 Cdc2(phospho-Cdc2 (Tyr15)) 단백질의 발현이 증가함을 확인하였다. #765를 처리하면 cyclinB1의 발현이 PANC-1에서는 증가하지만, Mia PaCa-2에서는 증가하지 않았다. 또한, 도 4b에 나타난 바와 같이, 유사분열 중단의 표지자로 알려진 Ser10번 잔기-인산화된 Histone H3(phospho-Histone H3 (Ser10)) 단백질이 #765를 처리하자 Mia PaCa-2와 PANC-1에서 발현이 증가함을 확인하였다. PANC-1에 #765를 처리하였을 때 phospho-Hisstone H3(Ser10)의 발현이 증가함을 면역형광염색을 이용하여 확인하였다(도 4c). 도 3과 도 4를 종합하면, #765가 Cdk1/cyclinB1 복합체의 활성화와 phospho-Histone H3 (Ser10)의 발현 증가를 유도함으로써 유사분열 중단을 야기한다는 것을 알 수 있다. To investigate whether the cell proliferation inhibitory effect of #765 is caused by the inhibition of cell cycle progression, the cell cycle progression of the pancreatic cancer cell line after treatment with #765 was confirmed through flow cytometry. As a result, as shown in FIG. 3 , it was observed that when cells were treated with #765, cells in G2/M stage were accumulated in both Mia PaCa-2 and PANC-1, and cells in G1 and S stage decreased. In addition, it can be seen that #765 induces cell death, as the number of cells in the subG1 stage was increased when #765 was treated with a high concentration. As a result of confirming the expression of the protein regulating the cell cycle using Western blot, as shown in FIG. 4A, when #765 was treated, Mia PaCa-2 and PANC-1 were involved in the progression from G2 to M phase. It was confirmed that the known Tyr15 residue-phosphorylated Cdc2 (phospho-Cdc2 (Tyr15)) protein expression was increased. Treatment with #765 increased the expression of cyclinB1 in PANC-1, but not Mia PaCa-2. In addition, as shown in FIG. 4b , the Ser10 residue-phospho-Histone H3 (Ser10) protein, known as a marker of mitotic disruption, was expressed in Mia PaCa-2 and PANC-1 when #765 was treated. It was confirmed that this increased. When PANC-1 was treated with #765, it was confirmed by immunofluorescence staining that the expression of phospho-Hisstone H3 (Ser10) was increased ( FIG. 4c ). Combining Figures 3 and 4, it can be seen that #765 induces the activation of the Cdk1/cyclinB1 complex and an increase in the expression of phospho-Histone H3 (Ser10), thereby causing mitotic disruption.
튜불린의 탈중합화 촉진 Promotes depolymerization of tubulin
#765가 유사분열 중단을 일으키는 기전을 밝히기 위해, 시험관 내 튜불린 중합 평가법(in vitro cell-free tubulin polymerization assay)을 이용하여 #765가 미세관 동역학(microtubule dynamics)에 미치는 영향을 알아보았다. 도 5a에서 나타난 바와 같이, 미세관 탈중합화 시약인 nocodazole과 마찬가지로 #765가 튜불린의 탈중합화를 일으켰다. 아울러, 면역형광염색법을 이용하여 PANC-1 세포에서 #765가 nocodazole과 마찬가지로 미세관을 붕괴시킴을 확인하였다(도 5b). 이를 종합하면, #765가 튜불린의 탈중합화를 유도하고, 이것이 인간 췌장암 세포의 유사분열 중단까지 일으킨다는 것을 알 수 있다.To elucidate the mechanism by which #765 causes mitotic disruption, the effect of #765 on microtubule dynamics was investigated using an in vitro cell-free tubulin polymerization assay. As shown in FIG. 5A , like nocodazole, a microtubule depolymerization reagent, #765 caused tubulin depolymerization. In addition, it was confirmed that #765 disrupted microtubules in PANC-1 cells like nocodazole using immunofluorescence staining (FIG. 5b). Taken together, it can be seen that #765 induces depolymerization of tubulin, which leads to mitotic disruption of human pancreatic cancer cells.
#765의 인간 췌장암 세포의 세포사멸 유도#765 Induction of apoptosis of human pancreatic cancer cells
Annexin-Ⅴ/7-AAD를 이용한 유세포 분석을 통해 #765가 세포사멸을 일으키는지 조사한 결과, 도 6에서 나타난 바와 같이, Mia PaCa-2와 PANC-1 세포에 #765를 처리하였을 때 농도 의존적으로 세포사멸이 일어남에 반하여 HPDE 세포에서는 세포사멸을 거의 일으키지 않았다. 이를 통해 #765가 정상세포가 아닌 췌장암 세포에 선택적인 세포독성을 보인다는 것을 알 수 있다. 세포사멸 관련 단백질의 발현을 웨스턴 블롯을 통해 확인한 결과, #765를 처리한 Mia PaCa-2와 PANC-1 세포에서 잘린 카스파아제-3(caspase-3)와 잘린 다중(에이디피-리보스) 당중합효소(PARP)의 발현이 증가함을 확인하였다(도 7a 및 7b). 아울러 #765의 처리는 항 세포사멸(항-apoptotic) 단백질인 Bcl-2의 발현을 감소시키고, 세포사멸을 일으키는(pro-apoptotic) 단백질인 Bim의 발현은 증가시켰다(도 7c). 종합하면, #765가 Mia PaCa-2와 PANC-1 세포에서 카스파아제-3와 다중(에이디피-리보스)당중합효소를 잘리게 함으로써 세포사멸을 유도한다는 것을 알 수 있다.As a result of investigating whether #765 causes apoptosis through flow cytometry analysis using Annexin-V/7-AAD, as shown in FIG. 6, when #765 was treated in Mia PaCa-2 and PANC-1 cells, concentration-dependently In contrast to apoptosis, HPDE cells rarely caused apoptosis. This shows that #765 shows selective cytotoxicity to pancreatic cancer cells rather than normal cells. As a result of confirming the expression of apoptosis-related proteins through western blot, cleaved caspase-3 and cleaved multiple (ADP-ribose) sugar polymerase in #765-treated Mia PaCa-2 and PANC-1 cells. It was confirmed that the expression of (PARP) is increased ( FIGS. 7a and 7b ). In addition, treatment with #765 decreased the expression of Bcl-2, an anti-apoptotic (anti-apoptotic) protein, and increased the expression of Bim, a pro-apoptotic protein ( FIG. 7c ). Taken together, it can be seen that #765 induces apoptosis by cleaving caspase-3 and poly(ADP-ribose) sugar polymerase in Mia PaCa-2 and PANC-1 cells.
#765에 의해 유도된 세포사멸에서의 JNK의 활성화Activation of JNK in apoptosis induced by #765
JNK 신호가 #765에 의해 유도된 세포사멸에 어떠한 역할을 하는지를 웨스턴블롯으로 분석하였다(도 8). 그 결과, #765를 처리한 Mia PaCa-2 세포에서 Thr183 잔기와 Tyr185 잔기(Thr183/Tyr185)에 인산화된 JNK의 발현이 시간과 농도 의존적으로 증가하였다(도 8a 및 8b). 이러한 JNK의 활성화가 #765에 의해서 일어나는 세포사멸에 필요한지를 조사하기 위해, JNK 특이적 억제제인 SP600125를 사용하였다. 도 8c에 나타난 바와 같이, SP600125를 #765 처리하기 2시간 전에 미리 처리한 경우, #765에 의해서 유도된 잘린 PARP의 발현이 다시 감소하였다. 이를 통해, JNK 신호가 #765에 의해 유도된 세포사멸과 관련되어있음을 확인하였다.The role of JNK signal in apoptosis induced by #765 was analyzed by Western blot (FIG. 8). As a result, in Mia PaCa-2 cells treated with #765, the expression of JNK phosphorylated at Thr183 residues and Tyr185 residues (Thr183/Tyr185) was increased in a time- and concentration-dependent manner ( FIGS. 8a and 8b ). To investigate whether this activation of JNK is necessary for apoptosis caused by #765, a JNK-specific inhibitor, SP600125, was used. As shown in FIG. 8c , when SP600125 was pretreated 2 hours before #765 treatment, the expression of truncated PARP induced by #765 decreased again. Through this, it was confirmed that the JNK signal is related to apoptosis induced by #765.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
Claims (10)
화학식 1
상기 화학식에서 R1은 NA1A2(A1 및 A2는 각각 독립적으로 수소 또는 C1-C3 알킬이다)이고 R2는 수소 또는 C1-C3 알킬이다.
A compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof:
Formula 1
In the above formula, R 1 is NA 1 A 2 (A 1 and A 2 are each independently hydrogen or C 1 -C 3 alkyl) and R 2 is hydrogen or C 1 -C 3 alkyl.
The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein A 1 and A 2 are hydrogen.
The compound or pharmaceutically acceptable salt thereof according to claim 1, wherein R 2 is C 1 alkyl.
A composition for preventing or treating cancer comprising the compound of any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof as an active ingredient.
The composition according to claim 4, wherein the cancer is selected from the group consisting of pancreatic cancer, breast cancer, colorectal cancer and gastric cancer.
The composition of claim 5, wherein the cancer is pancreatic cancer.
The composition according to claim 4, wherein the cancer is metastatic or recurrent cancer.
The composition according to claim 4, wherein the composition activates c-Jun N-terminal kinase (JNK).
화학식 1
상기 화학식에서 R1은 NA1A2(A1 및 A2는 각각 독립적으로 수소 또는 C1-C3 알킬이다)이고 R2는 수소 또는 C1-C3 알킬이다.
A functional food composition for the prevention or improvement of cancer comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
Formula 1
In the above formula, R 1 is NA 1 A 2 (A 1 and A 2 are each independently hydrogen or C 1 -C 3 alkyl) and R 2 is hydrogen or C 1 -C 3 alkyl.
화학식 1
상기 화학식에서 R1은 NA1A2(A1 및 A2는 각각 독립적으로 수소 또는 C1-C3 알킬이다)이고 R2는 수소 또는 C1-C3 알킬이다.A functional food composition for inhibiting the activity of LoxL2 (Lysyl oxidase like 2) comprising a compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof as an active ingredient:
Formula 1
In the above formula, R 1 is NA 1 A 2 (A 1 and A 2 are each independently hydrogen or C 1 -C 3 alkyl) and R 2 is hydrogen or C 1 -C 3 alkyl.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020210032023A KR102608183B1 (en) | 2021-03-11 | 2021-03-11 | Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same |
PCT/KR2022/003425 WO2022191656A1 (en) | 2021-03-11 | 2022-03-11 | Novel indolizine derivative and composition for prevention or treatment of cancer comprising same |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020210032023A KR102608183B1 (en) | 2021-03-11 | 2021-03-11 | Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same |
Publications (2)
Publication Number | Publication Date |
---|---|
KR20220127570A true KR20220127570A (en) | 2022-09-20 |
KR102608183B1 KR102608183B1 (en) | 2023-11-30 |
Family
ID=83228200
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
KR1020210032023A KR102608183B1 (en) | 2021-03-11 | 2021-03-11 | Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same |
Country Status (2)
Country | Link |
---|---|
KR (1) | KR102608183B1 (en) |
WO (1) | WO2022191656A1 (en) |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150028130A (en) * | 2013-09-05 | 2015-03-13 | 연세대학교 산학협력단 | Indolizine derivatives, pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition containing the same as an active ingredient |
KR20170125897A (en) * | 2015-03-06 | 2017-11-15 | 파마케아, 인크. | Fluorinated Lysyloxydase-like 2 inhibitors and uses thereof |
KR20190119269A (en) | 2018-04-12 | 2019-10-22 | 김영은 | Door Look System having Improved Crime Prevention Function Fireproof Function |
-
2021
- 2021-03-11 KR KR1020210032023A patent/KR102608183B1/en active IP Right Grant
-
2022
- 2022-03-11 WO PCT/KR2022/003425 patent/WO2022191656A1/en active Application Filing
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR20150028130A (en) * | 2013-09-05 | 2015-03-13 | 연세대학교 산학협력단 | Indolizine derivatives, pharmaceutically acceptable salt thereof, preparation method thereof and pharmaceutical composition containing the same as an active ingredient |
KR20170125897A (en) * | 2015-03-06 | 2017-11-15 | 파마케아, 인크. | Fluorinated Lysyloxydase-like 2 inhibitors and uses thereof |
KR20190119269A (en) | 2018-04-12 | 2019-10-22 | 김영은 | Door Look System having Improved Crime Prevention Function Fireproof Function |
Non-Patent Citations (5)
Title |
---|
Bioorg. Med. Chem. Lett., Vol. 26, No. 1, pp. 110-113 * |
Cell Res., Vol. 30, No. 10, pp. 885-901 * |
J. Org. Chem., Vol. 78, No. 20, pp. 10395-10404 * |
Mol. Cell Biol., Vol. 31, No. 16, pp. 3286-3297 * |
Oncotarget, Vol. 7, No. 27, pp. 42539-42552 * |
Also Published As
Publication number | Publication date |
---|---|
WO2022191656A1 (en) | 2022-09-15 |
KR102608183B1 (en) | 2023-11-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Luo et al. | Structure-activity relationships of 2, 4-disubstituted pyrimidines as dual ERα/VEGFR-2 ligands with anti-breast cancer activity | |
CN101743007A (en) | Inhibitors of akt/pkb with anti-tumor activity | |
TW201713328A (en) | Methods for treating cancer | |
KR20080107794A (en) | Composition comprising the extract of arctium lappa seed for anti-cancer | |
KR20140015798A (en) | Effective anticancer drugs against gefitinib-resistant human lung cancer cells comprising the daphnane diterpenoid compound | |
AU2018215809A1 (en) | N1 -(4-(5-(cyclopropylmethyl)-1 -methyl-1 H-pyrazol-4-yl)pyridin-2-yl)cyclohexane-1,4-diamine derivatives and related compounds as CK1 and/or IRAKI inhibitors for treating cancer | |
KR102608183B1 (en) | Novel Indolizine Derivatives and A Composition for Treating or Preventing Cancer Comprising the Same | |
KR20210146498A (en) | Pharmaceutical Composition for Preventing or Treating Cancer Comprising Inositol Polyphosphate Multikinase Inhibitor as an Active Ingredient | |
CN108473504A (en) | Novel dihydropyran hepyramine derivative and application thereof | |
CN108619145B (en) | Application of compound in treating tumor | |
KR101418161B1 (en) | Pharmaceutical compositions for prevention or treatment of liver cancer comprising neferine | |
US20090124647A1 (en) | Indole Derivatives as Antitumoral Compounds | |
KR102528732B1 (en) | Pharmaceutical composition for preventing or treating cancer comprising poziotinib and calcium channel blocker | |
KR101430310B1 (en) | Pharmaceutical composition for prevention or treatment of colorectal cancer comprising Juncus effecus L extract | |
Aziz et al. | Novel thiazolidines of potential anti-proliferation properties against esophageal squamous cell carcinoma via ERK pathway | |
KR20170089822A (en) | A composition for preventing or treating lung cancer comprising atranorin or salt thereof as an active ingredient | |
KR101699659B1 (en) | Composition preventing or treating cancer comprising (Z)-5-(4-hydroxy-3,5-dimethoxybenzylidene)-2-thioxoimidazolidin-4-one) | |
KR101941045B1 (en) | Pharmaceutical composition for preventing and treating cancer | |
KR20220147511A (en) | Composition for inhibiting cyclin-dependent kinase and medical uses thereof | |
KR102419029B1 (en) | Pharmaceutical composition for prevention and treatment of DYRK related diseases comprising phenanthrene-lactam compounds | |
KR20240041441A (en) | Composition for preventing or treating of breast cancer comprising compound from Dendropanax morbiferus | |
KR20230037853A (en) | Composition for preventing or treating pancreatic cancer | |
KR20230000108A (en) | Pharmaceutical composition for preventing or treating lung cancer comprising diethylstilbestrol | |
KR101890993B1 (en) | A diphenylnaphthylpyrazolinylcarbothioamide derivative with anti-cancer activity | |
KR101679604B1 (en) | New Compound Inhibiting Binding between DX2 and p14/ARF and Pharmaceutical Composition for Treating or Preventing Cancer Comprising the Same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
E902 | Notification of reason for refusal | ||
E701 | Decision to grant or registration of patent right | ||
GRNT | Written decision to grant |