JP5496232B2 - Il−8発現抑制剤 - Google Patents
Il−8発現抑制剤 Download PDFInfo
- Publication number
- JP5496232B2 JP5496232B2 JP2012046299A JP2012046299A JP5496232B2 JP 5496232 B2 JP5496232 B2 JP 5496232B2 JP 2012046299 A JP2012046299 A JP 2012046299A JP 2012046299 A JP2012046299 A JP 2012046299A JP 5496232 B2 JP5496232 B2 JP 5496232B2
- Authority
- JP
- Japan
- Prior art keywords
- rifampicin
- group
- expression
- gene
- serum
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 108090001007 Interleukin-8 Proteins 0.000 title claims description 108
- 230000014509 gene expression Effects 0.000 title claims description 107
- 102000004890 Interleukin-8 Human genes 0.000 title claims description 75
- 239000003112 inhibitor Substances 0.000 title claims description 27
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 claims description 139
- 229960001225 rifampicin Drugs 0.000 claims description 116
- -1 hydroxycarbonylmethylene group Chemical group 0.000 claims description 90
- 210000004027 cell Anatomy 0.000 claims description 41
- 239000001257 hydrogen Substances 0.000 claims description 22
- 229910052739 hydrogen Inorganic materials 0.000 claims description 22
- 150000003839 salts Chemical class 0.000 claims description 20
- 150000002431 hydrogen Chemical class 0.000 claims description 13
- BBNQHOMJRFAQBN-UPZFVJMDSA-N 3-formylrifamycin sv Chemical compound OC1=C(C(O)=C2C)C3=C(O)C(C=O)=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O BBNQHOMJRFAQBN-UPZFVJMDSA-N 0.000 claims description 12
- 206010073071 hepatocellular carcinoma Diseases 0.000 claims description 12
- HJYYPODYNSCCOU-ODRIEIDWSA-N rifamycin SV Chemical compound OC1=C(C(O)=C2C)C3=C(O)C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O HJYYPODYNSCCOU-ODRIEIDWSA-N 0.000 claims description 10
- 229940109171 rifamycin sv Drugs 0.000 claims description 10
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 9
- 125000004448 alkyl carbonyl group Chemical group 0.000 claims description 7
- 230000003115 biocidal effect Effects 0.000 claims description 6
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 claims description 6
- SQTCRTQCPJICLD-KTQDUKAHSA-N rifamycin B Chemical compound OC1=C(C(O)=C2C)C3=C(OCC(O)=O)C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O SQTCRTQCPJICLD-KTQDUKAHSA-N 0.000 claims description 6
- SQTCRTQCPJICLD-OQQFTUDCSA-N rifomycin-B Natural products COC1C=COC2(C)Oc3c(C)c(O)c4c(O)c(NC(=O)C(=C/C=C/C(C)C(O)C(C)C(O)C(C)C(OC(=O)C)C1C)C)cc(OCC(=O)O)c4c3C2=O SQTCRTQCPJICLD-OQQFTUDCSA-N 0.000 claims description 6
- 125000001424 substituent group Chemical group 0.000 claims description 6
- 229960002599 rifapentine Drugs 0.000 claims description 5
- WDZCUPBHRAEYDL-GZAUEHORSA-N rifapentine Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C(O)=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N(CC1)CCN1C1CCCC1 WDZCUPBHRAEYDL-GZAUEHORSA-N 0.000 claims description 5
- 125000000676 alkoxyimino group Chemical group 0.000 claims description 4
- 210000002950 fibroblast Anatomy 0.000 claims description 4
- XKTZWUACRZHVAN-VADRZIEHSA-N interleukin-8 Chemical compound C([C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@@H](NC(C)=O)CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCSC)C(=O)N1[C@H](CCC1)C(=O)N1[C@H](CCC1)C(=O)N[C@@H](C)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@H](CC(O)=O)C(=O)N[C@H](CC=1C=CC(O)=CC=1)C(=O)N[C@H](CO)C(=O)N1[C@H](CCC1)C(N)=O)C1=CC=CC=C1 XKTZWUACRZHVAN-VADRZIEHSA-N 0.000 description 69
- 229940096397 interleukin-8 Drugs 0.000 description 69
- 239000000243 solution Substances 0.000 description 42
- 239000002158 endotoxin Substances 0.000 description 25
- 229920006008 lipopolysaccharide Polymers 0.000 description 25
- 210000002966 serum Anatomy 0.000 description 25
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 24
- 108020004999 messenger RNA Proteins 0.000 description 19
- 230000002401 inhibitory effect Effects 0.000 description 17
- 238000000034 method Methods 0.000 description 16
- 108090000623 proteins and genes Proteins 0.000 description 12
- 230000000694 effects Effects 0.000 description 9
- 108010085238 Actins Proteins 0.000 description 8
- HJYYPODYNSCCOU-ZDHWWVNNSA-N Rifamycin SV Natural products COC1C=COC2(C)Oc3c(C)c(O)c4c(O)c(NC(=O)C(=C/C=C/C(C)C(O)C(C)C(O)C(C)C(OC(=O)C)C1C)C)cc(O)c4c3C2=O HJYYPODYNSCCOU-ZDHWWVNNSA-N 0.000 description 8
- 201000007270 liver cancer Diseases 0.000 description 8
- 208000014018 liver neoplasm Diseases 0.000 description 8
- 125000004674 methylcarbonyl group Chemical group CC(=O)* 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 230000001629 suppression Effects 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 230000009471 action Effects 0.000 description 6
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 238000003633 gene expression assay Methods 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 239000012679 serum free medium Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 125000006273 (C1-C3) alkyl group Chemical group 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 239000002585 base Substances 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000000338 in vitro Methods 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 238000003753 real-time PCR Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 description 3
- 239000002202 Polyethylene glycol Substances 0.000 description 3
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 3
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 3
- 239000004480 active ingredient Substances 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 210000000440 neutrophil Anatomy 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 229920001223 polyethylene glycol Polymers 0.000 description 3
- YVOFSHPIJOYKSH-NLYBMVFSSA-M sodium rifomycin sv Chemical compound [Na+].OC1=C(C(O)=C2C)C3=C([O-])C=C1NC(=O)\C(C)=C/C=C/[C@H](C)[C@H](O)[C@@H](C)[C@@H](O)[C@@H](C)[C@H](OC(C)=O)[C@H](C)[C@@H](OC)\C=C\O[C@@]1(C)OC2=C3C1=O YVOFSHPIJOYKSH-NLYBMVFSSA-M 0.000 description 3
- 229940124597 therapeutic agent Drugs 0.000 description 3
- 125000006552 (C3-C8) cycloalkyl group Chemical group 0.000 description 2
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 2
- 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 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 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 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 102100034459 Hepatitis A virus cellular receptor 1 Human genes 0.000 description 2
- 101710185991 Hepatitis A virus cellular receptor 1 homolog Proteins 0.000 description 2
- 101100020518 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) kyn-1 gene Proteins 0.000 description 2
- 101100288370 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) kyn-2 gene Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 125000000217 alkyl group Chemical group 0.000 description 2
- 239000003125 aqueous solvent Substances 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 239000011230 binding agent Substances 0.000 description 2
- 239000001768 carboxy methyl cellulose Substances 0.000 description 2
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 2
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 2
- QRYRORQUOLYVBU-VBKZILBWSA-N carnosic acid Chemical compound CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000002270 dispersing agent Substances 0.000 description 2
- 239000003995 emulsifying agent Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 150000004677 hydrates Chemical class 0.000 description 2
- 125000001841 imino group Chemical group [H]N=* 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000000314 lubricant Substances 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
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 238000007911 parenteral administration Methods 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000000375 suspending agent Substances 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
- 201000008827 tuberculosis Diseases 0.000 description 2
- 210000003556 vascular endothelial cell Anatomy 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- 125000000008 (C1-C10) alkyl group Chemical group 0.000 description 1
- 125000004398 2-methyl-2-butyl group Chemical group CC(C)(CC)* 0.000 description 1
- 125000004918 2-methyl-2-pentyl group Chemical group CC(C)(CCC)* 0.000 description 1
- 125000004922 2-methyl-3-pentyl group Chemical group CC(C)C(CC)* 0.000 description 1
- 125000004493 2-methylbut-1-yl group Chemical group CC(C*)CC 0.000 description 1
- 125000004917 3-methyl-2-butyl group Chemical group CC(C(C)*)C 0.000 description 1
- 125000004919 3-methyl-2-pentyl group Chemical group CC(C(C)*)CC 0.000 description 1
- 125000004921 3-methyl-3-pentyl group Chemical group CC(CC)(CC)* 0.000 description 1
- 125000004920 4-methyl-2-pentyl group Chemical group CC(CC(C)*)C 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 102000007469 Actins Human genes 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
- 239000005995 Aluminium silicate Substances 0.000 description 1
- 241001468213 Amycolatopsis mediterranei Species 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 108050006947 CXC Chemokine Proteins 0.000 description 1
- 102000019388 CXC chemokine Human genes 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 208000006154 Chronic hepatitis C Diseases 0.000 description 1
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- RGHNJXZEOKUKBD-SQOUGZDYSA-M D-gluconate Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O RGHNJXZEOKUKBD-SQOUGZDYSA-M 0.000 description 1
- 102000004163 DNA-directed RNA polymerases Human genes 0.000 description 1
- 108090000626 DNA-directed RNA polymerases Proteins 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
- 102000013138 Drug Receptors Human genes 0.000 description 1
- 108010065556 Drug Receptors Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 102100037813 Focal adhesion kinase 1 Human genes 0.000 description 1
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 1
- 208000005176 Hepatitis C Diseases 0.000 description 1
- 102100021866 Hepatocyte growth factor Human genes 0.000 description 1
- 101000878536 Homo sapiens Focal adhesion kinase 1 Proteins 0.000 description 1
- 101000898034 Homo sapiens Hepatocyte growth factor Proteins 0.000 description 1
- 101100508538 Homo sapiens IKBKE gene Proteins 0.000 description 1
- 101001046677 Homo sapiens Integrin alpha-V Proteins 0.000 description 1
- 101001015004 Homo sapiens Integrin beta-3 Proteins 0.000 description 1
- 101000599852 Homo sapiens Intercellular adhesion molecule 1 Proteins 0.000 description 1
- 101001033249 Homo sapiens Interleukin-1 beta Proteins 0.000 description 1
- 101000852992 Homo sapiens Interleukin-12 subunit beta Proteins 0.000 description 1
- 101001076408 Homo sapiens Interleukin-6 Proteins 0.000 description 1
- 101000973618 Homo sapiens NF-kappa-B essential modulator Proteins 0.000 description 1
- 101000961071 Homo sapiens NF-kappa-B inhibitor alpha Proteins 0.000 description 1
- 101001128158 Homo sapiens Nanos homolog 2 Proteins 0.000 description 1
- 101001124991 Homo sapiens Nitric oxide synthase, inducible Proteins 0.000 description 1
- 101000868152 Homo sapiens Son of sevenless homolog 1 Proteins 0.000 description 1
- 101000611183 Homo sapiens Tumor necrosis factor Proteins 0.000 description 1
- 101000622304 Homo sapiens Vascular cell adhesion protein 1 Proteins 0.000 description 1
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-N Hydrogen bromide Chemical compound Br CPELXLSAUQHCOX-UHFFFAOYSA-N 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 102000001284 I-kappa-B kinase Human genes 0.000 description 1
- 108060006678 I-kappa-B kinase Proteins 0.000 description 1
- 108091058560 IL8 Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 102100021857 Inhibitor of nuclear factor kappa-B kinase subunit epsilon Human genes 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 102100022337 Integrin alpha-V Human genes 0.000 description 1
- 102100032999 Integrin beta-3 Human genes 0.000 description 1
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102100039065 Interleukin-1 beta Human genes 0.000 description 1
- 102100036701 Interleukin-12 subunit beta Human genes 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 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
- 206010024229 Leprosy Diseases 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 1
- 102100022219 NF-kappa-B essential modulator Human genes 0.000 description 1
- 102100039337 NF-kappa-B inhibitor alpha Human genes 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 1
- 108020005497 Nuclear hormone receptor Proteins 0.000 description 1
- 102000007399 Nuclear hormone receptor Human genes 0.000 description 1
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 1
- 101150044441 PECAM1 gene Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 235000019483 Peanut oil Nutrition 0.000 description 1
- 239000004264 Petrolatum Substances 0.000 description 1
- 229920002675 Polyoxyl Polymers 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 102000000804 Pregnane X Receptor Human genes 0.000 description 1
- 108010001511 Pregnane X Receptor Proteins 0.000 description 1
- 239000004820 Pressure-sensitive adhesive Substances 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 230000006819 RNA synthesis Effects 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 229930189077 Rifamycin Natural products 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 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 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 1
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical class CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 108010053096 Vascular Endothelial Growth Factor Receptor-1 Proteins 0.000 description 1
- 108010053099 Vascular Endothelial Growth Factor Receptor-2 Proteins 0.000 description 1
- 102100023543 Vascular cell adhesion protein 1 Human genes 0.000 description 1
- 102100039037 Vascular endothelial growth factor A Human genes 0.000 description 1
- 102100033178 Vascular endothelial growth factor receptor 1 Human genes 0.000 description 1
- 102100033177 Vascular endothelial growth factor receptor 2 Human genes 0.000 description 1
- YKTSYUJCYHOUJP-UHFFFAOYSA-N [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] Chemical compound [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] YKTSYUJCYHOUJP-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 239000000556 agonist Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 229910052783 alkali metal Inorganic materials 0.000 description 1
- 229910052784 alkaline earth metal Inorganic materials 0.000 description 1
- 235000012211 aluminium silicate Nutrition 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000033115 angiogenesis Effects 0.000 description 1
- 239000002870 angiogenesis inducing agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 1
- 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 1
- 230000004071 biological effect Effects 0.000 description 1
- 159000000007 calcium salts Chemical class 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000003399 chemotactic effect Effects 0.000 description 1
- 239000005482 chemotactic factor Substances 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000002285 corn oil Substances 0.000 description 1
- 235000005687 corn oil Nutrition 0.000 description 1
- 235000012343 cottonseed oil Nutrition 0.000 description 1
- 239000002385 cottonseed oil Substances 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 125000001995 cyclobutyl group Chemical group [H]C1([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 125000000582 cycloheptyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- 125000000113 cyclohexyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C1([H])[H] 0.000 description 1
- 125000000640 cyclooctyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])([H])C([H])(*)C([H])([H])C([H])([H])C1([H])[H] 0.000 description 1
- 125000001511 cyclopentyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 1
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- VILAVOFMIJHSJA-UHFFFAOYSA-N dicarbon monoxide Chemical group [C]=C=O VILAVOFMIJHSJA-UHFFFAOYSA-N 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000975 dye Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 150000002169 ethanolamines Chemical class 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 125000004672 ethylcarbonyl group Chemical group [H]C([H])([H])C([H])([H])C(*)=O 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229940050410 gluconate Drugs 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 229940116364 hard fat Drugs 0.000 description 1
- 208000010710 hepatitis C virus infection Diseases 0.000 description 1
- XMBWDFGMSWQBCA-UHFFFAOYSA-N hydrogen iodide Chemical compound I XMBWDFGMSWQBCA-UHFFFAOYSA-N 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 239000001866 hydroxypropyl methyl cellulose Substances 0.000 description 1
- 235000010979 hydroxypropyl methyl cellulose Nutrition 0.000 description 1
- 229920003088 hydroxypropyl methyl cellulose Polymers 0.000 description 1
- UFVKGYZPFZQRLF-UHFFFAOYSA-N hydroxypropyl methyl cellulose Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC2C(C(O)C(OC3C(C(O)C(O)C(CO)O3)O)C(CO)O2)O)C(CO)O1 UFVKGYZPFZQRLF-UHFFFAOYSA-N 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 150000007529 inorganic bases Chemical class 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 description 1
- 125000001972 isopentyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- NLYAJNPCOHFWQQ-UHFFFAOYSA-N kaolin Chemical compound O.O.O=[Al]O[Si](=O)O[Si](=O)O[Al]=O NLYAJNPCOHFWQQ-UHFFFAOYSA-N 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 159000000003 magnesium salts Chemical class 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
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000037323 metabolic rate Effects 0.000 description 1
- 150000003956 methylamines Chemical class 0.000 description 1
- 125000001570 methylene group Chemical group [H]C([H])([*:1])[*:2] 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 239000003226 mitogen Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000001280 n-hexyl group Chemical group C(CCCCC)* 0.000 description 1
- 125000000740 n-pentyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 108020004017 nuclear receptors Proteins 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 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
- 125000003544 oxime group Chemical group 0.000 description 1
- 125000004430 oxygen atom Chemical group O* 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000000312 peanut oil Substances 0.000 description 1
- 125000003538 pentan-3-yl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])C([H])([H])[H] 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 229940066842 petrolatum Drugs 0.000 description 1
- 235000019271 petrolatum Nutrition 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 108010001062 polysaccharide-K Proteins 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- XAEFZNCEHLXOMS-UHFFFAOYSA-M potassium benzoate Chemical compound [K+].[O-]C(=O)C1=CC=CC=C1 XAEFZNCEHLXOMS-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000001023 pro-angiogenic effect Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 229960003292 rifamycin Drugs 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 125000002914 sec-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 239000008159 sesame oil Substances 0.000 description 1
- 235000011803 sesame oil Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 1
- 239000000725 suspension 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
- 229940095064 tartrate Drugs 0.000 description 1
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- OGIDPMRJRNCKJF-UHFFFAOYSA-N titanium oxide Inorganic materials [Ti]=O OGIDPMRJRNCKJF-UHFFFAOYSA-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
Landscapes
- Nitrogen And Oxygen Or Sulfur-Condensed Heterocyclic Ring Systems (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
IL−8が過剰発現することに起因する疾患の予防・治療方法や、IL−8が過剰発現することに起因する疾患の予防・治療方法に使用するためのリファンピシン等なども含まれる。
肝がん細胞におけるリファンピシンの作用を調べるために、肝がん細胞株にリファンピシンを添加し、遺伝子発現レベルを解析した。
[1]肝がん細胞株(SK−HEP−1)を1.0x104/cm2程度の密度で、6ウェルプレートに播種し、10%FBS含むDEME(ナカライテスク社製)又はRPMI1640(ナカライテスク社製)(以下、「血清含有培養液」という)存在下で24時間培養した。
[2]血清含有培養液又は血清不含のOPTI−MEM(GibcoBRL社製)(以下、「血清不含培養液」という)のそれぞれに、リファンピシン溶液(50mg/ml又は25mg/ml in DMSO)を1000倍希釈になるように加え、かかるリファンピシン(25μg/ml又は50μg/ml)を含む血清含有又は血清不含の培養液を、工程[1]の血清含有培養液と置換した。なお、上記リファンピシン溶液は、和光純薬社製の粉末リファンピシン試薬を、和光純薬製のDMSOで当該濃度に溶解して調製した。
[3]インキュベータ内(37℃、5%CO2条件下)で5時間培養を行った後、細胞を回収し、RNAの精製を行った。RNAの精製は、RNeasy Plus Mini Kit(Qiagen社製)を用いて製品付属のプロトコールにしたがって行った。なお、コントロールとしてリファンピシンを含まない血清含有又は血清不含の培養液で培養した細胞のRNAを用いた。ただし、コントロールに用いたリファンピシン不含培養液にも、リファンピシン含有培養液と同濃度のDMSOが含まれるようにDMSOを添加した。
[1]以下の1)〜4)からなる反応液を調製した。
1)20ng/μlRNA;2.0μl
2)2×QuantiFast Probe RT-PCR Master Mix(Qiagen社製);10μl
3)TaqMan Gene Expression Assayプローブ・プライマーセット(Applied Biosystems社製);1.0μl
4)RNaseフリーの脱イオン水;7.0μl
5)Reverse Transcriptase;0.1μl
1)50℃、20分を1サイクル(mRNAからcDNAへの逆転写反応)
2)95℃、5分を1サイクル(ポリメラーゼの活性化)
3)95℃と60℃との往復を40サイクル(TaqMan Gene Expression Assayプローブ・プライマーセットによるcDNAの増幅)
[3]Baselineソフトウェア(Applied Biosystems社製)を用いてPCR産物が一定量になるPCRのサイクル数(threshold cycle;Ct値)を測定し、比較Ct法(ΔΔCt法)によりβアクチン遺伝子PCR産物のCt値を基準とした上記21種類の遺伝子PCR産物のCt値の相対値を求め、かかるCt値の相対値から、上記21種類の遺伝子由来のPCR産物の相対量、すなわち上記21種類の遺伝子のmRNAの相対量を算出した。
リアルタイムRT−PCRによって、リファンピシン添加による上記21種類の遺伝子の、SK−HEP−1細胞中での発現量の変動を解析した結果、リファンピシンを添加しない場合と比べて、リファンピシンを添加した場合、IL−8遺伝子のmRNAの発現量は2倍抑制されることが明らかとなった。なお、IL−8以外の他の20種類の遺伝子については、リファンピシン添加による有意なmRNAの発現量の変動は認められなかった。また、血清含有培養液を用いた場合と比べて、血清不含培養液を用いた場合、リファンピシンによるIL−8遺伝子の発現抑制効果は2.5倍高かった。リファンピシンの濃度が血清含有及び血清不含培養液で同じであるにもかかわらず、血清不含培養液でリファンピシンによるIL−8遺伝子の発現抑制効果が高かった理由としては、例えば培養液中に含まれる血清がリファンピシンの細胞内への浸透性を低下させたため、血清含有培養液においては、リファンピシンによるIL−8遺伝子の発現抑制効果が抑制されたことが考えられる。
がん細胞の中にはIL−8遺伝子の発現が恒常的に亢進しているものが知られている。そこで、リファンピシンによるIL−8遺伝子の発現抑制効果が、かかる細胞においても認められるかどうかについて解析を行った。まず、2種類の正常線維芽細胞株(TIG−3、WI−38)と、11種類の肝がん細胞株(HuH−7、Hep−3B、Hep G2、Li−7、SK−HEP−1、KIM−1、KYN−1、KYN−2、KYN−3、HAK−1A、及びHAK−1B)におけるIL−8遺伝子のmRNAの発現量について、上記実施例1記載の方法により解析を行った(図1)。その結果、4種類の肝がん細胞株(HuH−7、Hep−3B、Li−7、及びSK−HEP−1)について、IL−8遺伝子の発現が恒常的に亢進していることが認められた(図1)。その中で特にIL−8遺伝子発現量が高かったHep−3B及びSK−HEP−1細胞株を用いて、上記実施例1に記載の方法によりIL−8遺伝子の発現量を解析したところ、Hep−3B細胞を用いた場合も、SK−HEP−1細胞を用いた場合と同様に、IL−8遺伝子の発現抑制効果が認められた。すなわち、Hep−3B細胞株を用いた場合、リファンピシン(25μg/ml又は50μg/ml)を含む血清含有培養液を用いたときには約30%、リファンピシン(25μg/ml又は50μg/ml)を含む血清不含培養液を用いたときには約80%のIL−8遺伝子の発現を抑制することが明らかとなった(図2の左図)。また、SK−HEP−1細胞株を用いた場合、リファンピシン25μg/ml又は50μg/ml)を含む血清含有培養液を用いたときには約50%、リファンピシン25μg/ml又は50μg/ml)を含む血清不含培養液を用いたときには約80%のIL−8遺伝子の発現を抑制することが明らかとなった(図2の右図)。これらの結果は、IL−8遺伝子の発現が恒常的に亢進している細胞においても、リファンピシンによるIL−8遺伝子の発現抑制効果が認められることを示している。
さらに、IL−8遺伝子の発現量が少ない細胞においても、LPSによりIL−8遺伝子の発現量を亢進させた場合、同様にリファンピシンによるIL−8遺伝子発現抑制効果が認められるかどうかを解析した。IL−8遺伝子の発現量が少ない細胞株として、上記実施例2で解析した13種類の肝がん細胞株の中から、IL−8遺伝子の発現量が少ない3種類の肝がん細胞株(Hep G2、KYN−3、及びHAK−1B)を選択した。なお、LPSを含む血清不含培養液を用いてかかる3種類の肝がん細胞株をLPS存在下で培養したところ、LPSによるIL−8遺伝子発現の亢進はほとんど認められなかったため、血清含有培養液について解析を進めた。血清含有培養液に、LPS(Lipopolysaccharides from E.coli 055:B5、SIGMA社製)(1mg/ml in PBS)及びリファンピシン溶液(和光純薬社製)(25mg/ml in DMSO)をそれぞれ1000倍希釈になるように加え、かかるLPS(1μg/ml)及びリファンピシン(25μg/ml)を含む血清含有培養液(「LPS+RFP」)を用いて、上記3種類の肝がん細胞株(Hep G2、KYN−3、及びHAK−1B)を5時間培養した。また、コントロールとして、血清含有培養液(「NT」)や、LPS(1μg/ml)を含む血清含有培養液(「LPS」)を用いた。なお、リファンピシンを含まないこれらのコントロール培養液(上記「NT」や、「LPS」)には、リファンピシン含有培養液(上記「LPS+RFP」)と同濃度のDMSOが含まれるようにDMSOを添加した。
次に、リファンピシンの誘導体についてもリファンピシンと同様のIL−8遺伝子の発現抑制効果があるかどうかについて解析を行った。リファンピシンの誘導体として、リファマイシンSVナトリウム塩及び3−フォルミルリファマイシンSVナトリウム塩を用い、また、肝がん細胞株として、SK−HEP−1細胞株を用いて上記実施例1に記載の方法によりIL−8遺伝子の発現量を解析したところ、リファンピシンを用いた場合と同様に、IL−8遺伝子の発現抑制効果が認められた。すなわち、リファマイシンSV(25μg/ml)を含む血清含有及び血清不含の培養液を用いた場合、それぞれ約50%及び約80%のIL−8遺伝子の発現を抑制できることが明らかとなった(図4の左図)。また、3−フォルミルリファマイシンSV(25μg/ml)を含む血清含有及び血清不含の培養液を用いた場合、それぞれ約50%のIL−8遺伝子の発現を抑制できることが明らかとなった(図4の右図)。これらの結果は、リファンピシンの誘導体についてもリファンピシンと同様のIL−8遺伝子の発現抑制効果があることを示している。
次に、肝がん細胞株(SK−HEP−1)を用いて、IL−8遺伝子の発現抑制する、リファンピシンの作用濃度について検討を行った。リファンピシンを含む血清含有又は血清不含の培養液として、1、3、10、30、又は100(μg/ml)のリファンピシン濃度のものを用いて上記実施例1に記載の方法によりIL−8遺伝子の発現量を解析したところ、リファンピシンを含む血清含有の培養液を用いた場合、リファンピシン濃度3μg/mlでIL−8遺伝子の発現量は約50%にまで減少しており、この結果は、リファンピシン濃度が1〜3μg/mlの間に50%のIL−8遺伝子の発現を抑制する濃度(IC50;Half maximal [50%] inhibitory concentration)値があることを示している(図5の左図)。また、リファンピシンを含む血清含有の培養液を用いた場合、少なくとも3μg/mlのリファンピシン濃度でIL−8遺伝子の発現を約50%抑制できることが明らかとなった(図5の左図)。一方、リファンピシンを含む血清不含の培養液を用いた場合、リファンピシン濃度1μg/mlでIL−8遺伝子の発現量は50%以下にまで減少しており、この結果はリファンピシン濃度が0〜1μg/mlの間にIC50値があることを示している(図5の右図)。また、リファンピシンを含む血清不含の培養液を用いた場合、少なくとも3μg/mlのリファンピシン濃度でIL−8遺伝子の発現を約80%抑制できることが明らかとなった(図5の右図)。
次に、肝がん細胞株(SK−HEP−1)を用いて、IL−8遺伝子の発現を抑制する、リファンピシンの作用時間について検討を行った。リファンピシンを含む血清含有又は血清不含の培養液として、10(μg/ml)のリファンピシン濃度の血清不含の培養液を用い、0、10、30、60、180、又は300分間それぞれインキュベータ内(37℃、5%CO2条件下)でSK−HEP−1細胞株を上記培養液存在下で培養を行い、上記実施例1に記載の方法によりL−8遺伝子の発現量を行ったところ、リファンピシン添加後60〜180分の間にIL−8遺伝子のmRNAの発現量が減少することが認められた(図6)。この結果は、リファンピシンによるIL−8遺伝子の発現抑制効果は、少なくともリファンピシン添加後60〜180分の間に認められることを示している。また、リファンピシンによるIL−8遺伝子の発現抑制効果に即効性があることから、IL−8遺伝子のmRNAの代謝速度は速いことが予想される。
Claims (2)
- 正常線維芽細胞と比較してIL−8遺伝子の発現が高い肝がん細胞に対するIL−8発現抑制剤であって、
リファンピシン、リファンピシン誘導体、リファンピシンの薬理学的に許容しうる塩、又は、リファンピシン誘導体の薬理学的に許容しうる塩を含有し、
前記リファンピシン又はリファンピシン誘導体が、以下の一般式(I)で示されるリファンピシン系抗生物質である、IL−8発現抑制剤。
- リファンピシン系抗生物質が、リファンピシン、リファマイシン−SV、3−フォルミルリファマイシン、リファペンチン、リファマイシンBからなる群より選ばれる請求項1に記載のIL−8発現抑制剤。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2012046299A JP5496232B2 (ja) | 2012-03-02 | 2012-03-02 | Il−8発現抑制剤 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2012046299A JP5496232B2 (ja) | 2012-03-02 | 2012-03-02 | Il−8発現抑制剤 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2013180992A JP2013180992A (ja) | 2013-09-12 |
JP5496232B2 true JP5496232B2 (ja) | 2014-05-21 |
Family
ID=49271909
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2012046299A Active JP5496232B2 (ja) | 2012-03-02 | 2012-03-02 | Il−8発現抑制剤 |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP5496232B2 (ja) |
-
2012
- 2012-03-02 JP JP2012046299A patent/JP5496232B2/ja active Active
Also Published As
Publication number | Publication date |
---|---|
JP2013180992A (ja) | 2013-09-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11370763B2 (en) | Compounds and compositions for treating conditions associated with NLRP activity | |
US20220339169A1 (en) | Methods of treating or selecting a treatment for a subject resistant to tnf inhibitor using a nlrp3 antagonist | |
JP6625631B2 (ja) | 1本鎖rnaウイルス複製を阻害するための治療 | |
US9572801B2 (en) | Tetrahydronaphthyridinyl propionic acid derivatives and uses thereof | |
US20180110796A1 (en) | Compositions and methods for the treatment of hbv infection | |
JP2022537570A (ja) | がんを処置する方法 | |
JP2024502755A (ja) | がんを処置する方法 | |
JP5496232B2 (ja) | Il−8発現抑制剤 | |
JP6852122B2 (ja) | Cyr61及びvegf媒介性の状態の治療 | |
US20210330605A1 (en) | Anti-cancer activity of adamantane derivatives | |
AU2013214731B2 (en) | Compositions comprising NDGA derivatives and sorafenib and their use in treatment of cancer | |
WO2022140387A1 (en) | Methods of treating cancer | |
JP2024500874A (ja) | がんを処置する方法 | |
JP2024500877A (ja) | がんを処置する方法 | |
JP5061352B2 (ja) | 肝臓疾患治療剤及び肝機能改善剤 | |
WO2017163243A1 (en) | Modulation of calcium channel splice variant in cancer therapy | |
EP2829536A1 (en) | 4-Nitro-5-dichloromethylpyrazol derivatives for the treatment of infectious diseases | |
JPWO2020004404A1 (ja) | IL−1β阻害薬 | |
WO2020055946A1 (en) | Targeting egln1 in cancer | |
JP5424960B2 (ja) | Ccr2bまたはccr5とフロントタンパク質との相互作用の阻害剤 | |
US20240131047A1 (en) | Small molecule inhibitors of cd38 as immunosuppressants | |
WO2023171789A1 (ja) | Vla-4の阻害物質を含む、心筋炎を予防、又は治療するための組成物 | |
JP2010173956A (ja) | ピロール化合物を有効成分として含有するケモカイン産生阻害剤 | |
JPH11222474A (ja) | 糖尿病治療薬 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20130926 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20131120 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20140206 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20140304 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 5496232 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S531 | Written request for registration of change of domicile |
Free format text: JAPANESE INTERMEDIATE CODE: R313531 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |