JP2009013126A - Prion protein structure transformation inhibitor - Google Patents
Prion protein structure transformation inhibitor Download PDFInfo
- Publication number
- JP2009013126A JP2009013126A JP2007178247A JP2007178247A JP2009013126A JP 2009013126 A JP2009013126 A JP 2009013126A JP 2007178247 A JP2007178247 A JP 2007178247A JP 2007178247 A JP2007178247 A JP 2007178247A JP 2009013126 A JP2009013126 A JP 2009013126A
- Authority
- JP
- Japan
- Prior art keywords
- group
- compound
- substituted
- prion
- prion protein
- 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.)
- Pending
Links
- 108091000054 Prion Proteins 0.000 title claims abstract description 105
- 239000003112 inhibitor Substances 0.000 title claims abstract description 12
- 102000029797 Prion Human genes 0.000 title abstract description 101
- 230000009466 transformation Effects 0.000 title abstract description 8
- 150000001875 compounds Chemical class 0.000 claims abstract description 158
- 208000024777 Prion disease Diseases 0.000 claims abstract description 26
- 208000015181 infectious disease Diseases 0.000 claims abstract description 26
- 230000002458 infectious effect Effects 0.000 claims abstract description 24
- 125000005843 halogen group Chemical group 0.000 claims abstract description 15
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 9
- 125000001183 hydrocarbyl group Chemical group 0.000 claims abstract 7
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 37
- 238000006243 chemical reaction Methods 0.000 claims description 31
- 125000001424 substituent group Chemical group 0.000 claims description 31
- 229910052717 sulfur Inorganic materials 0.000 claims description 24
- 239000003814 drug Substances 0.000 claims description 19
- 238000003032 molecular docking Methods 0.000 claims description 19
- 239000002253 acid Substances 0.000 claims description 18
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 claims description 17
- 238000004088 simulation Methods 0.000 claims description 17
- 239000011593 sulfur Substances 0.000 claims description 17
- 125000000623 heterocyclic group Chemical group 0.000 claims description 16
- 125000002252 acyl group Chemical group 0.000 claims description 14
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 14
- 125000003277 amino group Chemical group 0.000 claims description 13
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 claims description 13
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 13
- 239000004480 active ingredient Substances 0.000 claims description 12
- 229910052757 nitrogen Inorganic materials 0.000 claims description 12
- 125000004433 nitrogen atom Chemical group N* 0.000 claims description 12
- 125000001820 oxy group Chemical group [*:1]O[*:2] 0.000 claims description 12
- 125000004149 thio group Chemical group *S* 0.000 claims description 12
- 229940124597 therapeutic agent Drugs 0.000 claims description 11
- 125000005740 oxycarbonyl group Chemical group [*:1]OC([*:2])=O 0.000 claims description 10
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 10
- 102100034452 Alternative prion protein Human genes 0.000 claims description 9
- 201000010099 disease Diseases 0.000 claims description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 8
- 125000004185 ester group Chemical group 0.000 claims description 8
- RTZKZFJDLAIYFH-UHFFFAOYSA-N ether Substances CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 8
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 claims description 8
- 125000004430 oxygen atom Chemical group O* 0.000 claims description 8
- 125000004093 cyano group Chemical group *C#N 0.000 claims description 7
- 125000004434 sulfur atom Chemical group 0.000 claims description 7
- 230000003449 preventive effect Effects 0.000 claims description 6
- 241001494479 Pecora Species 0.000 claims description 5
- 208000008864 scrapie Diseases 0.000 claims description 5
- 208000018756 Variant Creutzfeldt-Jakob disease Diseases 0.000 claims description 4
- 208000005881 bovine spongiform encephalopathy Diseases 0.000 claims description 4
- 125000005647 linker group Chemical group 0.000 claims description 4
- 208000020406 Creutzfeldt Jacob disease Diseases 0.000 claims description 3
- 208000003407 Creutzfeldt-Jakob Syndrome Diseases 0.000 claims description 3
- 208000010859 Creutzfeldt-Jakob disease Diseases 0.000 claims description 3
- 206010023497 kuru Diseases 0.000 claims description 2
- 230000006806 disease prevention Effects 0.000 claims 1
- -1 (substituted) nitrogen Chemical class 0.000 abstract description 33
- 239000003795 chemical substances by application Substances 0.000 abstract description 9
- 230000000069 prophylactic effect Effects 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 239000000203 mixture Substances 0.000 abstract description 2
- 238000000034 method Methods 0.000 description 43
- 210000004027 cell Anatomy 0.000 description 18
- 150000002430 hydrocarbons Chemical group 0.000 description 17
- 238000012360 testing method Methods 0.000 description 17
- 238000012216 screening Methods 0.000 description 15
- 230000000694 effects Effects 0.000 description 14
- 235000018102 proteins Nutrition 0.000 description 14
- 230000000144 pharmacologic effect Effects 0.000 description 13
- RWRDLPDLKQPQOW-UHFFFAOYSA-N Pyrrolidine Chemical compound C1CCNC1 RWRDLPDLKQPQOW-UHFFFAOYSA-N 0.000 description 12
- 102000004169 proteins and genes Human genes 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 10
- 125000001931 aliphatic group Chemical group 0.000 description 10
- 238000004519 manufacturing process Methods 0.000 description 10
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 9
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 9
- 125000006239 protecting group Chemical group 0.000 description 9
- 239000000126 substance Substances 0.000 description 9
- 102100025818 Major prion protein Human genes 0.000 description 8
- 229940079593 drug Drugs 0.000 description 8
- 238000000126 in silico method Methods 0.000 description 7
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 6
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 description 6
- 125000002723 alicyclic group Chemical group 0.000 description 6
- 229940024606 amino acid Drugs 0.000 description 6
- 235000001014 amino acid Nutrition 0.000 description 6
- 150000001413 amino acids Chemical class 0.000 description 6
- 238000001514 detection method Methods 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 239000002904 solvent Substances 0.000 description 6
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 5
- 238000005481 NMR spectroscopy Methods 0.000 description 5
- 125000002029 aromatic hydrocarbon group Chemical group 0.000 description 5
- 238000011156 evaluation Methods 0.000 description 5
- 230000003405 preventing effect Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- KDCGOANMDULRCW-UHFFFAOYSA-N 7H-purine Chemical compound N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 4
- YLQBMQCUIZJEEH-UHFFFAOYSA-N Furan Chemical compound C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 4
- 101100298534 Mus musculus Prnp gene Proteins 0.000 description 4
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 4
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical compound C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 description 4
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 4
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 4
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 4
- 125000000217 alkyl group Chemical group 0.000 description 4
- 125000006615 aromatic heterocyclic group Chemical group 0.000 description 4
- 125000004432 carbon atom Chemical group C* 0.000 description 4
- 238000004440 column chromatography Methods 0.000 description 4
- 125000005842 heteroatom Chemical group 0.000 description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- KZNICNPSHKQLFF-UHFFFAOYSA-N succinimide Chemical group O=C1CCC(=O)N1 KZNICNPSHKQLFF-UHFFFAOYSA-N 0.000 description 4
- ZWEHNKRNPOVVGH-UHFFFAOYSA-N 2-Butanone Chemical compound CCC(C)=O ZWEHNKRNPOVVGH-UHFFFAOYSA-N 0.000 description 3
- YNNKKMKHXXUFLP-UHFFFAOYSA-N 2-pyrrolidin-1-yl-n-[4-[[4-[(2-pyrrolidin-1-ylacetyl)amino]phenyl]methyl]phenyl]acetamide Chemical compound C=1C=C(CC=2C=CC(NC(=O)CN3CCCC3)=CC=2)C=CC=1NC(=O)CN1CCCC1 YNNKKMKHXXUFLP-UHFFFAOYSA-N 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- UNLIFCVLZKVZQK-UHFFFAOYSA-N CC(C(=O)NC1=CC=C(C=C1)CC2=CC=C(C=C2)NC(=O)CN3CCCC3)N4CCCC4 Chemical compound CC(C(=O)NC1=CC=C(C=C1)CC2=CC=C(C=C2)NC(=O)CN3CCCC3)N4CCCC4 UNLIFCVLZKVZQK-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 125000004423 acyloxy group Chemical group 0.000 description 3
- 125000003118 aryl group Chemical group 0.000 description 3
- 125000004429 atom Chemical group 0.000 description 3
- JFDZBHWFFUWGJE-UHFFFAOYSA-N benzonitrile Chemical compound N#CC1=CC=CC=C1 JFDZBHWFFUWGJE-UHFFFAOYSA-N 0.000 description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 3
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 3
- 125000000753 cycloalkyl group Chemical group 0.000 description 3
- 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 3
- 238000012217 deletion Methods 0.000 description 3
- 230000037430 deletion Effects 0.000 description 3
- 239000000543 intermediate Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 125000006700 (C1-C6) alkylthio group Chemical group 0.000 description 2
- SCYULBFZEHDVBN-UHFFFAOYSA-N 1,1-Dichloroethane Chemical compound CC(Cl)Cl SCYULBFZEHDVBN-UHFFFAOYSA-N 0.000 description 2
- FLBAYUMRQUHISI-UHFFFAOYSA-N 1,8-naphthyridine Chemical compound N1=CC=CC2=CC=CN=C21 FLBAYUMRQUHISI-UHFFFAOYSA-N 0.000 description 2
- FCEHBMOGCRZNNI-UHFFFAOYSA-N 1-benzothiophene Chemical group C1=CC=C2SC=CC2=C1 FCEHBMOGCRZNNI-UHFFFAOYSA-N 0.000 description 2
- IPXNXMNCBXHYLQ-UHFFFAOYSA-N 2-pyrrolidin-1-ylacetic acid Chemical compound OC(=O)CN1CCCC1 IPXNXMNCBXHYLQ-UHFFFAOYSA-N 0.000 description 2
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical group [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 2
- PAYRUJLWNCNPSJ-UHFFFAOYSA-N Aniline Chemical compound NC1=CC=CC=C1 PAYRUJLWNCNPSJ-UHFFFAOYSA-N 0.000 description 2
- 241000283690 Bos taurus Species 0.000 description 2
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 2
- 239000004215 Carbon black (E152) Substances 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- YNQLUTRBYVCPMQ-UHFFFAOYSA-N Ethylbenzene Chemical compound CCC1=CC=CC=C1 YNQLUTRBYVCPMQ-UHFFFAOYSA-N 0.000 description 2
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- PEEHTFAAVSWFBL-UHFFFAOYSA-N Maleimide Chemical group O=C1NC(=O)C=C1 PEEHTFAAVSWFBL-UHFFFAOYSA-N 0.000 description 2
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 description 2
- PCNDJXKNXGMECE-UHFFFAOYSA-N Phenazine Natural products C1=CC=CC2=NC3=CC=CC=C3N=C21 PCNDJXKNXGMECE-UHFFFAOYSA-N 0.000 description 2
- WTKZEGDFNFYCGP-UHFFFAOYSA-N Pyrazole Chemical compound C=1C=NNC=1 WTKZEGDFNFYCGP-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- YTPLMLYBLZKORZ-UHFFFAOYSA-N Thiophene Chemical compound C=1C=CSC=1 YTPLMLYBLZKORZ-UHFFFAOYSA-N 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 125000002339 acetoacetyl group Chemical group O=C([*])C([H])([H])C(=O)C([H])([H])[H] 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- 125000005035 acylthio group Chemical group 0.000 description 2
- 125000004453 alkoxycarbonyl group Chemical group 0.000 description 2
- 230000000389 anti-prion effect Effects 0.000 description 2
- 125000004104 aryloxy group Chemical group 0.000 description 2
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 2
- 229910052794 bromium Inorganic materials 0.000 description 2
- 238000004422 calculation algorithm Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 125000000392 cycloalkenyl group Chemical group 0.000 description 2
- 125000005170 cycloalkyloxycarbonyl group Chemical group 0.000 description 2
- 125000005366 cycloalkylthio group Chemical group 0.000 description 2
- 125000001995 cyclobutyl group Chemical group [H]C1([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 2
- 125000000596 cyclohexenyl group Chemical group C1(=CCCCC1)* 0.000 description 2
- 125000001511 cyclopentyl group Chemical group [H]C1([H])C([H])([H])C([H])([H])C([H])(*)C1([H])[H] 0.000 description 2
- 125000001559 cyclopropyl group Chemical group [H]C1([H])C([H])([H])C1([H])* 0.000 description 2
- 125000002704 decyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 230000002554 disease preventive effect Effects 0.000 description 2
- 229910052731 fluorine Inorganic materials 0.000 description 2
- 239000011737 fluorine Substances 0.000 description 2
- 125000002485 formyl group Chemical group [H]C(*)=O 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- 229930195733 hydrocarbon Natural products 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 238000010191 image analysis Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 2
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 description 2
- 150000002611 lead compounds Chemical class 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 125000002950 monocyclic group Chemical group 0.000 description 2
- 125000006574 non-aromatic ring group Chemical group 0.000 description 2
- 125000004043 oxo group Chemical group O=* 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 125000001501 propionyl group Chemical group O=C([*])C([H])([H])C([H])([H])[H] 0.000 description 2
- PBMFSQRYOILNGV-UHFFFAOYSA-N pyridazine Chemical compound C1=CC=NN=C1 PBMFSQRYOILNGV-UHFFFAOYSA-N 0.000 description 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 2
- 125000002112 pyrrolidino group Chemical group [*]N1C([H])([H])C([H])([H])C([H])([H])C1([H])[H] 0.000 description 2
- JWVCLYRUEFBMGU-UHFFFAOYSA-N quinazoline Chemical compound N1=CN=CC2=CC=CC=C21 JWVCLYRUEFBMGU-UHFFFAOYSA-N 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 229960002317 succinimide Drugs 0.000 description 2
- 125000001036 sulfinic acid ester group Chemical group 0.000 description 2
- 125000000626 sulfinic acid group Chemical group 0.000 description 2
- 125000002130 sulfonic acid ester group Chemical group 0.000 description 2
- 125000000542 sulfonic acid group Chemical group 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- DYHSDKLCOJIUFX-UHFFFAOYSA-N tert-butoxycarbonyl anhydride Chemical compound CC(C)(C)OC(=O)OC(=O)OC(C)(C)C DYHSDKLCOJIUFX-UHFFFAOYSA-N 0.000 description 2
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 2
- BVQMUAAELWHQSF-UHFFFAOYSA-N tert-butyl n-[4-[(4-aminophenyl)methyl]phenyl]carbamate Chemical compound C1=CC(NC(=O)OC(C)(C)C)=CC=C1CC1=CC=C(N)C=C1 BVQMUAAELWHQSF-UHFFFAOYSA-N 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 150000003852 triazoles Chemical class 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- QWXZOFZKSQXPDC-NSHDSACASA-N (2s)-2-(9h-fluoren-9-ylmethoxycarbonylamino)propanoic acid Chemical compound C1=CC=C2C(COC(=O)N[C@@H](C)C(O)=O)C3=CC=CC=C3C2=C1 QWXZOFZKSQXPDC-NSHDSACASA-N 0.000 description 1
- 125000006673 (C1-C12) aliphatic hydrocarbon group Chemical group 0.000 description 1
- 125000004178 (C1-C4) alkyl group Chemical group 0.000 description 1
- 125000004191 (C1-C6) alkoxy group Chemical group 0.000 description 1
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- 125000005837 1,2-cyclopentylene group Chemical group [H]C1([H])C([H])([H])C([H])([*:1])C([H])([*:2])C1([H])[H] 0.000 description 1
- 125000005838 1,3-cyclopentylene group Chemical group [H]C1([H])C([H])([H])C([H])([*:2])C([H])([H])C1([H])[*:1] 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- 125000004955 1,4-cyclohexylene group Chemical group [H]C1([H])C([H])([H])C([H])([*:1])C([H])([H])C([H])([H])C1([H])[*:2] 0.000 description 1
- ULTHEAFYOOPTTB-UHFFFAOYSA-N 1,4-dibromobutane Chemical compound BrCCCCBr ULTHEAFYOOPTTB-UHFFFAOYSA-N 0.000 description 1
- 125000004343 1-phenylethyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])(*)C([H])([H])[H] 0.000 description 1
- 125000000530 1-propynyl group Chemical group [H]C([H])([H])C#C* 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 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
- UXGVMFHEKMGWMA-UHFFFAOYSA-N 2-benzofuran Chemical compound C1=CC=CC2=COC=C21 UXGVMFHEKMGWMA-UHFFFAOYSA-N 0.000 description 1
- 125000000094 2-phenylethyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 description 1
- 125000006201 3-phenylpropyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 125000000590 4-methylphenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)C([H])([H])[H] 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 1
- DKPFZGUDAPQIHT-UHFFFAOYSA-N Butyl acetate Natural products CCCCOC(C)=O DKPFZGUDAPQIHT-UHFFFAOYSA-N 0.000 description 1
- LTCFXYLUCOVGMH-UHFFFAOYSA-N CC(C)(C)OC(Nc1ccc(Cc(cc2)ccc2NC(C(C)(C)N2CCCC2)=O)cc1)=O Chemical compound CC(C)(C)OC(Nc1ccc(Cc(cc2)ccc2NC(C(C)(C)N2CCCC2)=O)cc1)=O LTCFXYLUCOVGMH-UHFFFAOYSA-N 0.000 description 1
- 0 CCCCCCCN[C@@](C)(*)C1OC1O Chemical compound CCCCCCCN[C@@](C)(*)C1OC1O 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 description 1
- XTHFKEDIFFGKHM-UHFFFAOYSA-N Dimethoxyethane Chemical compound COCCOC XTHFKEDIFFGKHM-UHFFFAOYSA-N 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 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
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 241000283086 Equidae Species 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- FXHOOIRPVKKKFG-UHFFFAOYSA-N N,N-Dimethylacetamide Chemical compound CN(C)C(C)=O FXHOOIRPVKKKFG-UHFFFAOYSA-N 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- ZCQWOFVYLHDMMC-UHFFFAOYSA-N Oxazole Chemical compound C1=COC=N1 ZCQWOFVYLHDMMC-UHFFFAOYSA-N 0.000 description 1
- WYNCHZVNFNFDNH-UHFFFAOYSA-N Oxazolidine Chemical compound C1COCN1 WYNCHZVNFNFDNH-UHFFFAOYSA-N 0.000 description 1
- 239000002033 PVDF binder Substances 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
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- XBDQKXXYIPTUBI-UHFFFAOYSA-M Propionate Chemical compound CCC([O-])=O XBDQKXXYIPTUBI-UHFFFAOYSA-M 0.000 description 1
- CVQUWLDCFXOXEN-UHFFFAOYSA-N Pyran-4-one Chemical compound O=C1C=COC=C1 CVQUWLDCFXOXEN-UHFFFAOYSA-N 0.000 description 1
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical group C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 1
- DHXVGJBLRPWPCS-UHFFFAOYSA-N Tetrahydropyran Chemical compound C1CCOCC1 DHXVGJBLRPWPCS-UHFFFAOYSA-N 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- KXKVLQRXCPHEJC-UHFFFAOYSA-N acetic acid trimethyl ester Natural products COC(C)=O KXKVLQRXCPHEJC-UHFFFAOYSA-N 0.000 description 1
- ORILYTVJVMAKLC-UHFFFAOYSA-N adamantane Chemical group C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 description 1
- 125000005073 adamantyl group Chemical group C12(CC3CC(CC(C1)C3)C2)* 0.000 description 1
- 239000003463 adsorbent Substances 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 229960003767 alanine Drugs 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 150000001338 aliphatic hydrocarbons Chemical class 0.000 description 1
- 125000003342 alkenyl group Chemical group 0.000 description 1
- 125000003545 alkoxy group Chemical group 0.000 description 1
- 125000002947 alkylene group Chemical group 0.000 description 1
- 125000000304 alkynyl group Chemical group 0.000 description 1
- HSFWRNGVRCDJHI-UHFFFAOYSA-N alpha-acetylene Natural products C#C HSFWRNGVRCDJHI-UHFFFAOYSA-N 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 229940064004 antiseptic throat preparations Drugs 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- 125000005098 aryl alkoxy carbonyl group Chemical group 0.000 description 1
- 125000003710 aryl alkyl group Chemical group 0.000 description 1
- 125000002102 aryl alkyloxo group Chemical group 0.000 description 1
- 125000005161 aryl oxy carbonyl group Chemical group 0.000 description 1
- 125000005110 aryl thio group Chemical group 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- METKIMKYRPQLGS-UHFFFAOYSA-N atenolol Chemical compound CC(C)NCC(O)COC1=CC=C(CC(N)=O)C=C1 METKIMKYRPQLGS-UHFFFAOYSA-N 0.000 description 1
- RFRXIWQYSOIBDI-UHFFFAOYSA-N benzarone Chemical compound CCC=1OC2=CC=CC=C2C=1C(=O)C1=CC=C(O)C=C1 RFRXIWQYSOIBDI-UHFFFAOYSA-N 0.000 description 1
- 125000001584 benzyloxycarbonyl group Chemical group C(=O)(OCC1=CC=CC=C1)* 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000006172 buffering agent Substances 0.000 description 1
- 125000004106 butoxy group Chemical group [*]OC([H])([H])C([H])([H])C(C([H])([H])[H])([H])[H] 0.000 description 1
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 125000003739 carbamimidoyl group Chemical group C(N)(=N)* 0.000 description 1
- 125000002837 carbocyclic group Chemical group 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- VZWXIQHBIQLMPN-UHFFFAOYSA-N chromane Chemical compound C1=CC=C2CCCOC2=C1 VZWXIQHBIQLMPN-UHFFFAOYSA-N 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- OTAFHZMPRISVEM-UHFFFAOYSA-N chromone Chemical compound C1=CC=C2C(=O)C=COC2=C1 OTAFHZMPRISVEM-UHFFFAOYSA-N 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000006482 condensation reaction Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 125000000000 cycloalkoxy group Chemical group 0.000 description 1
- 125000004210 cyclohexylmethyl group Chemical group [H]C([H])(*)C1([H])C([H])([H])C([H])([H])C([H])([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
- 125000002433 cyclopentenyl group Chemical group C1(=CCCC1)* 0.000 description 1
- 125000004851 cyclopentylmethyl group Chemical group C1(CCCC1)C* 0.000 description 1
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound 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 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 125000001664 diethylamino group Chemical group [H]C([H])([H])C([H])([H])N(*)C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 150000002170 ethers Chemical class 0.000 description 1
- 125000003754 ethoxycarbonyl group Chemical group C(=O)(OCC)* 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 125000004705 ethylthio group Chemical group C(C)S* 0.000 description 1
- 125000002534 ethynyl group Chemical group [H]C#C* 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003889 eye drop Substances 0.000 description 1
- 229940012356 eye drops Drugs 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 125000002795 guanidino group Chemical group C(N)(=N)N* 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- FUZZWVXGSFPDMH-UHFFFAOYSA-N hexanoic acid Chemical compound CCCCCC(O)=O FUZZWVXGSFPDMH-UHFFFAOYSA-N 0.000 description 1
- 125000004051 hexyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 238000002000 high resolution fast-atom bombardment mass spectrometry Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000002267 hypothalamic effect Effects 0.000 description 1
- 125000002883 imidazolyl group Chemical group 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000012750 in vivo screening Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 125000003387 indolinyl group Chemical group N1(CCC2=CC=CC=C12)* 0.000 description 1
- 125000001041 indolyl group Chemical group 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000007914 intraventricular administration Methods 0.000 description 1
- 229910052740 iodine Inorganic materials 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
- HEBMCVBCEDMUOF-UHFFFAOYSA-N isochromane Chemical compound C1=CC=C2COCCC2=C1 HEBMCVBCEDMUOF-UHFFFAOYSA-N 0.000 description 1
- JMMWKPVZQRWMSS-UHFFFAOYSA-N isopropanol acetate Natural products CC(C)OC(C)=O JMMWKPVZQRWMSS-UHFFFAOYSA-N 0.000 description 1
- 229940011051 isopropyl acetate Drugs 0.000 description 1
- ZLTPDFXIESTBQG-UHFFFAOYSA-N isothiazole Chemical compound C=1C=NSC=1 ZLTPDFXIESTBQG-UHFFFAOYSA-N 0.000 description 1
- GWYFCOCPABKNJV-UHFFFAOYSA-N isovaleric acid Chemical compound CC(C)CC(O)=O GWYFCOCPABKNJV-UHFFFAOYSA-N 0.000 description 1
- CTAPFRYPJLPFDF-UHFFFAOYSA-N isoxazole Chemical compound C=1C=NOC=1 CTAPFRYPJLPFDF-UHFFFAOYSA-N 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 125000001160 methoxycarbonyl group Chemical group [H]C([H])([H])OC(*)=O 0.000 description 1
- 125000002816 methylsulfanyl group Chemical group [H]C([H])([H])S[*] 0.000 description 1
- 238000000302 molecular modelling Methods 0.000 description 1
- 125000004573 morpholin-4-yl group Chemical group N1(CCOCC1)* 0.000 description 1
- 125000001421 myristyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 125000001624 naphthyl group Chemical group 0.000 description 1
- 239000007923 nasal drop Substances 0.000 description 1
- 229940100662 nasal drops Drugs 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 208000015122 neurodegenerative disease Diseases 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- UMRZSTCPUPJPOJ-KNVOCYPGSA-N norbornane Chemical group C1C[C@H]2CC[C@@H]1C2 UMRZSTCPUPJPOJ-KNVOCYPGSA-N 0.000 description 1
- 125000003518 norbornenyl group Chemical group C12(C=CC(CC1)C2)* 0.000 description 1
- 125000002868 norbornyl group Chemical group C12(CCC(CC1)C2)* 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 125000003261 o-tolyl group Chemical group [H]C1=C([H])C(*)=C(C([H])=C1[H])C([H])([H])[H] 0.000 description 1
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229950000964 pepstatin Drugs 0.000 description 1
- 108010091212 pepstatin Proteins 0.000 description 1
- FAXGPCHRFPCXOO-LXTPJMTPSA-N pepstatin A Chemical compound OC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)C[C@H](O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)CC(C)C FAXGPCHRFPCXOO-LXTPJMTPSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 description 1
- 125000006678 phenoxycarbonyl group Chemical group 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- 125000003356 phenylsulfanyl group Chemical group [*]SC1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 125000003367 polycyclic group Chemical group 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- FVSKHRXBFJPNKK-UHFFFAOYSA-N propionitrile Chemical compound CCC#N FVSKHRXBFJPNKK-UHFFFAOYSA-N 0.000 description 1
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000006333 protein structural change Effects 0.000 description 1
- HNJBEVLQSNELDL-UHFFFAOYSA-N pyrrolidin-2-one Chemical group O=C1CCCN1 HNJBEVLQSNELDL-UHFFFAOYSA-N 0.000 description 1
- 125000000168 pyrrolyl group Chemical group 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000004007 reversed phase HPLC 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
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000000859 sublimation Methods 0.000 description 1
- 230000008022 sublimation Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- BUUPQKDIAURBJP-UHFFFAOYSA-N sulfinic acid Chemical compound OS=O BUUPQKDIAURBJP-UHFFFAOYSA-N 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 125000005931 tert-butyloxycarbonyl group Chemical group [H]C([H])([H])C(OC(*)=O)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- VLLMWSRANPNYQX-UHFFFAOYSA-N thiadiazole Chemical compound C1=CSN=N1.C1=CSN=N1 VLLMWSRANPNYQX-UHFFFAOYSA-N 0.000 description 1
- 229930192474 thiophene Natural products 0.000 description 1
- VNXUJPCYZSNXDG-UHFFFAOYSA-N thiopyran-4-one Chemical group O=C1C=CSC=C1 VNXUJPCYZSNXDG-UHFFFAOYSA-N 0.000 description 1
- 125000003944 tolyl group Chemical group 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 125000003258 trimethylene group Chemical group [H]C([H])([*:2])C([H])([H])C([H])([H])[*:1] 0.000 description 1
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 238000003041 virtual screening Methods 0.000 description 1
- 238000005406 washing Methods 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
- 239000008096 xylene Substances 0.000 description 1
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
Description
本発明は、正常型プリオンタンパク質の結合ポケットへの結合を介して感染型プリオンの生成を抑制することができる新規な化合物、当該化合物を有効成分として含むプリオンタンパク質構造変換抑制剤、プリオン病の予防・治療剤、及び前記化合物を用いてプリオンタンパク質の構造変換抑制活性を有する化合物をスクリーニングする方法に関する。 The present invention relates to a novel compound capable of suppressing the production of infectious prion through binding to a binding pocket of normal prion protein, a prion protein structure conversion inhibitor containing the compound as an active ingredient, and prevention of prion disease -It is related with the therapeutic agent and the method of screening the compound which has the structure conversion inhibitory activity of prion protein using the said compound.
プリオン病とは、プリオンタンパク質の異常によって引き起こされる疾患の総称であって、例えば、羊のスクレイピー、ウシ海綿状脳症、クロイツフェルト・ヤコブ病、GSS、FFI、クールー(S.B.Prusiner, Science, 216, 136-144 (1982)、S.B.Prusiner, Science, 252, 1515-1522(1991)、S.B.Prusiner, Prions. Proc.Natl.Acad.Sci.U.S.A., 95, 13363-13383 (1998))、変異型ヤコブ病(G.Chazot et al., Lancet, 347, 1181-1181(1996)、R.G.Will et al., Lancet, 347, 921-925(1996))などの神経変性疾患などが知られている。このようなプリオン病を治療する方法として、従来、放射線、煮沸、化学薬品等を用いてプリオンを不活化する方法や、種々の手段により感染力を減衰させる方法などが試みられているが、プリオンの耐性は強く必ずしも効果があがっていなかった。 Prion disease is a general term for diseases caused by prion protein abnormalities. For example, sheep scrapie, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, GSS, FFI, Kool (SBPrusiner, Science, 216, 136 -144 (1982), SBPrusiner, Science, 252, 1515-1522 (1991), SBPrusiner, Prions. Proc. Natl. Acad. Sci. USA, 95, 13363-13383 (1998)), mutant Jacob disease ( Neurodegenerative diseases such as G. Chazot et al., Lancet, 347, 1181-1181 (1996) and RGWill et al., Lancet, 347, 921-925 (1996)) are known. As methods for treating such prion diseases, methods such as inactivation of prions using radiation, boiling, chemicals, etc., and methods of attenuating infectivity by various means have been tried. The tolerance of was not necessarily effective.
ここで、プリオン病は、プリオンタンパク質の異常に起因する疾患であって、具体的には、正常型プリオン(PrPC)が構造変換し、アミロイド前駆体特殊構造(PrP*)を経由して感染型プリオン(PrPSc)を生成し、感染型プリオンが体内で正常型プリオンと結合して、正常型プリオンは感染型プリオンに次々に変換されて症状が進行すると考えられている。(S.B.Prusiner, Science, 216, 136-144(1982)、S.B.Prusiner, Science, 252, 1515-1522(1991)、K.Kuwata et al., Biochemistry 41, 12277-12283(2002))。V.Perrier et al., Proc.Natl.Acad.Sci.U.S.A. 97, 6073-6078(2000)には、正常型構造のプリオンタンパク質に結合して構造変換を阻害する活性を有する化合物を抗プリオン薬の開発に利用することが示唆されているが、その薬理効果は 証されていない。 Here, prion disease is a disease caused by abnormality of prion protein. Specifically, normal prion (PrP C ) undergoes structural transformation and is infected via amyloid precursor special structure (PrP *). It is thought that the type prion (PrP Sc ) is generated, and the infectious type prion binds with the normal type prion in the body, and the normal type prion is successively converted into the infectious type prion and the symptoms progress. (SBPrusiner, Science, 216, 136-144 (1982), SBPrusiner, Science, 252, 1515-1522 (1991), K. Kuuwata et al., Biochemistry 41, 12277-12283 (2002)). In V. Perrier et al., Proc. Natl. Acad. Sci. USA 97, 6073-6078 (2000), an anti-prion drug is a compound that binds to a prion protein having a normal type structure and inhibits structural conversion. However, its pharmacological effect has not been proven.
現在まで、マウス(R.Riek et al., Nature, 382, 180-182 (1996))、ハムスター(T.L.James et al., Proc.Natl.Acad.Sci.U.S.A. 94, 10086-10091(1997))、ウシ(F.Lopez Garcia, R.Zahn, R.Riek, K.Wuthrick, Proc.Natl.Acad.Sci.U.S.A. 97, 8334-8339(2000))、ヒト(L.Calzolai et al.,Proc.Natl.Acad.Sci.U.S.A.97, 8340-8345(2000))に関してNMRによる三次元構造決定が行われている。これらの構造は、かなり良く似ていて、アミノ酸残基第128番から第231番までは、αへリックスからなるまとまった構造と不完全な柔らかいβシートからなる。N端から113番までは構造を取らず、113番から128番までは疎水性のクラスターを形成しており、複数の構造を形成している(H.Liu et al., Biochemistry 38, 5362-5377(1999))。また、これまで、プリオンタンパク質の構造変換に関連して、プリオン中間体が観測されている(H.Zhang et al., Biochemistry 36, 3543-53(1997)、S.Hornemann, R.A.Glockshuber, Proc.Natl.Acad.Sci.U.S.A. 95, 6010-6014(1998)、A.C.Apetri, W.K.Surewicz, Biol.Chem. 277, 44589-44592(2002))。近年、高圧NMR法(K.Kuwata et al., Biochemistry 41, 12277-12283(2002))により、プリオンのフォールディング中間体PrP*が同定され、その構造の特徴が明らかになった。PrP*においてはヘリックスBとCが変性しているが、へリックスAは保たれている。また、PrP*は生理的条件下で1%存在していることが分かっている(K.Kuwata et al.,Biochemistry 41, 12277-12283(2002))。 To date, mice (R. Riek et al., Nature, 382, 180-182 (1996)), hamsters (TLJames et al., Proc. Natl. Acad. Sci. USA 94, 10086-10091 (1997)) Cattle (F. Lopez Garcia, R. Zahn, R. Riek, K. Wuthrick, Proc. Natl. Acad. Sci. USA 97, 8334-8339 (2000)), humans (L. Calzolai et al., Proc. Natl. Acad. Sci. USA 97, 8340-8345 (2000)), three-dimensional structure determination by NMR is performed. These structures are quite similar, and amino acid residues No. 128 to No. 231 are composed of a structure composed of an α helix and an incomplete soft β sheet. No structure is formed from the N end to the 113th, and a hydrophobic cluster is formed from the 113th to the 128th, forming a plurality of structures (H. Liu et al., Biochemistry 38, 5362- 5377 (1999)). Until now, prion intermediates have been observed in relation to the structural transformation of prion protein (H. Zhang et al., Biochemistry 36, 3543-53 (1997), S. Hornemann, RAGlockshuber, Proc. Natl. Acad. Sci. USA 95, 6010-6014 (1998), ACApetri, WKSurewicz, Biol. Chem. 277, 44589-44592 (2002)). Recently, the prion folding intermediate PrP * has been identified and characterized by high-pressure NMR (K. Kuuwata et al., Biochemistry 41, 12277-12283 (2002)). In PrP *, helices B and C are denatured, but helix A is retained. PrP * is found to be present at 1% under physiological conditions (K. Kuuwata et al., Biochemistry 41, 12277-12283 (2002)).
上記知見に基づき、本発明者らは、正常型プリオンの構造体におけるA-S 2ループとヘリックスBのC端側との間にある結合ポケットを焦点に当てたドッキングシミュレーションにより、正常型プリオンタンパク質との結合を介して構造変換を抑制し、フォールディング中間体PrP*の生成を強く抑制する化合物のスクリーニングを行い、そのような化合物として2-pyrrolidin-1-yl-N-[4-[4-(2-pyrrolidin-1-yl-acetylamino)-benzyl]-phenyl]-acetamideを得ている(特開2005−120002号公報)。しかし、この化合物は、正常型プリオンと結合して構造変換を抑制しうるが、感染型プリオンの生成量の低減により優れた効果を発揮しうる化合物が求められていた。 Based on the above findings, the present inventors conducted a docking simulation focusing on the binding pocket between the AS 2 loop in the normal prion structure and the C-terminal side of helix B, and the normal prion protein. We screened for compounds that suppress the structural transformation through binding and strongly suppress the formation of the folding intermediate PrP *. As such compounds, 2-pyrrolidin-1-yl-N- [4- [4- (2 -pyrrolidin-1-yl-acetylamino) -benzyl] -phenyl] -acetamide has been obtained (Japanese Patent Laid-Open No. 2005-120002). However, this compound can bind to normal prions to suppress structural conversion, but there has been a demand for a compound that can exhibit superior effects by reducing the amount of infectious prions produced.
本発明の目的は、正常型プリオンタンパク質への結合を介して感染型プリオンタンパク質の生成を効率よく抑制することができる化合物、当該化合物を含むプリオンタンパク質構造変換抑制剤、及びプリオン病の予防・治療剤を提供することにある。
本発明の他の目的は、プリオンタンパク質の構造変換抑制効果に優れた化合物を効率よくスクリーニングする方法を提供することにある。
An object of the present invention is to provide a compound capable of efficiently suppressing the production of infectious prion protein through binding to normal prion protein, a prion protein structure conversion inhibitor containing the compound, and prevention / treatment of prion disease It is to provide an agent.
Another object of the present invention is to provide a method for efficiently screening for a compound excellent in the effect of suppressing the structural transformation of prion protein.
本発明者らは、上記目的を達成するため鋭意検討した結果、特定の構造を有する化合物によれば、プリオンタンパク質へ強固に結合することにより、当該プリオンタンパク質の正常型から感染型への構造変換を効率よく抑制し、プリオン病の発病や進行の防止に優れた効果を発揮することを見出し、本発明を完成した。 As a result of intensive studies to achieve the above object, the present inventors have found that, according to a compound having a specific structure, the prion protein is structurally converted from a normal form to an infectious form by firmly binding to the prion protein. The present invention has been completed by finding that it effectively suppresses the above and exerts an excellent effect in preventing the onset and progression of prion diseases.
すなわち、本発明は、下記式(1)
(式中、R1〜R4は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、又は置換若しくは無置換アミノ基を示す。R5〜R12は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、又は置換若しくは無置換アミノ基を示す。Xは単結合又は連結基を示す。環Z1及び環Z2は、それぞれ置換基を有していてもよい窒素原子含有環を示す。但し、R1〜R4の少なくとも一つは水素原子以外の基を示す)
で表される化合物を提供する。
That is, the present invention provides the following formula (1):
(Wherein R 1 to R 4 are the same or different and are a hydrogen atom, a halogen atom, an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, carboxyl Group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercapto group, substituted thio group, or substituted or unsubstituted R 5 to R 12 are the same or different and each represents a hydrogen atom, a halogen atom, a hydrocarbon group which may have a substituent, or a heterocyclic group which may have a substituent. , Carboxyl group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercap Group, a substituted thio group, or .X showing a substituted or unsubstituted amino group is a single bond or a linking group. Ring Z 1 and the ring Z 2 is a good nitrogen atom-containing ring which may have a substituent Provided that at least one of R 1 to R 4 represents a group other than a hydrogen atom)
The compound represented by these is provided.
前記式(1)中、好ましくは、R1〜R4は、水素原子、又は置換基を有していてもよい炭化水素基であり、R5〜R12は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、ヒドロキシル基、置換オキシ基、メルカプト基、又は置換チオ基であり、XはC1〜C4アルキレン基、酸素原子(エーテル結合;−O−)、又は硫黄原子(チオエーテル結合;−S−)であり、環Z1及び環Z2は、同一又は異なって、置換基を有していてもよい5員環又は6員環の窒素原子含有環であって、R1〜R4のうち2つが水素原子で2つが水素原子以外の基、又はR1〜R4のうち3つが水素原子で1つが水素原子以外の基である。特に好ましくは、下記式(2)
で表される化合物が用いられる。
In the formula (1), preferably, R 1 to R 4 are hydrogen atoms or optionally substituted hydrocarbon groups, and R 5 to R 12 are the same or different and are hydrogen atoms. , halogen atom, or an optionally substituted hydrocarbon group, a hydroxyl group, a substituted oxy group, a mercapto group, or a substituted thio group, X is C 1 -C 4 alkylene group, an oxygen atom (ether bond; -O-), or a sulfur atom (thioether bond; -S-), and ring Z 1 and ring Z 2 are the same or different and each may be a 5-membered or 6-membered ring optionally having a substituent. A nitrogen atom-containing ring, two of R 1 to R 4 are hydrogen atoms and two are groups other than hydrogen atoms, or three of R 1 to R 4 are hydrogen atoms and one is a group other than hydrogen atoms . Particularly preferably, the following formula (2)
The compound represented by these is used.
また、本発明は、上記本発明の化合物を有効成分として含むプリオンタンパク質構造変換抑制剤を提供する。 The present invention also provides a prion protein structure conversion inhibitor containing the compound of the present invention as an active ingredient.
さらに、本発明は、上記本発明の化合物を有効成分として含むプリオン病の予防・治療剤を提供する。前記プリオン病には、例えば、羊のスクレイピー、ウシ海綿状脳症、クロイツフェルト・ヤコブ病、GSS、FFI、クールーおよび変異型ヤコブ病等が含まれる。 Furthermore, the present invention provides a preventive / therapeutic agent for prion diseases comprising the compound of the present invention as an active ingredient. Examples of the prion disease include sheep scrapie, bovine spongiform encephalopathy, Creutzfeldt-Jakob disease, GSS, FFI, Kool, and mutant Jacob disease.
本発明は、さらに、上記本発明の化合物及び/又はその誘導体と、プリオンに感染した細胞と、感染型プリオンタンパク質を検出する手段とを含むキットを提供する。 The present invention further provides a kit comprising the compound of the present invention and / or a derivative thereof, a prion-infected cell, and a means for detecting an infectious prion protein.
また、本発明は、上記本発明の化合物及び/又はその誘導体を用い、正常型プリオンタンパク質とのドッキングシミュレーションにより予測される結合強さに基づき、プリオンタンパク質の構造変換抑制活性を有する化合物をスクリーニングする方法を提供する。 The present invention also uses the above-described compound of the present invention and / or a derivative thereof to screen for a compound having prion protein structure conversion inhibitory activity based on the binding strength predicted by docking simulation with a normal prion protein. Provide a method.
本願明細書中、「正常型プリオンタンパク質」とは、正常な細胞に発現している感染性を有しないプリオンタンパク質を意味しており、「感染型プリオンタンパク質」とは、正常型プリオンタンパク質とアミノ酸配列は同一であるが、立体構造が異なり、感染性を有するプリオンタンパク質を意味している。また、「プリオンに感染する」とは、感染型プリオンタンパク質に感染している状態を意味している。 In the present specification, “normal prion protein” means a prion protein that is expressed in normal cells and has no infectivity, and “infectious prion protein” means normal prion protein and amino acid. It means a prion protein having the same sequence but different steric structure and having infectivity. Further, “infecting prion” means a state of being infected with infectious prion protein.
本発明の化合物は、プリオンタンパク質と強固に結合するため、当該プリオンタンパク質の構造変換を抑制する効果に極めて優れている。このような効果を奏する化合物は、正常型プリオンの構造変換に起因する感染型プリオンの生成を抑制する作用を奏するプリオンタンパク質構造変換抑制剤として有用である。本発明の化合物は、上記作用を奏するため、プリオン病の発症や症状の進行の防止効果に優れたプリオン病の予防・治療剤を提供することができる。本発明のスクリーニング方法によれば、上記化合物を用いるため、プリオンタンパク質の構造変換を抑制する効果に優れた化合物を効率よく得ることができる。 Since the compound of the present invention binds firmly to the prion protein, it is extremely excellent in the effect of suppressing the structural conversion of the prion protein. A compound that exhibits such an effect is useful as a prion protein structural conversion inhibitor that has an effect of suppressing the production of infectious prions resulting from the structural conversion of normal prions. Since the compound of the present invention exerts the above-described action, it can provide a preventive / therapeutic agent for prion disease that is excellent in preventing the onset of prion disease and the progression of symptoms. According to the screening method of the present invention, since the above compound is used, a compound excellent in the effect of suppressing the structural transformation of the prion protein can be obtained efficiently.
本発明の化合物は、前記式(1)で表される。式(1)中、R1〜R4は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、又は置換若しくは無置換アミノ基を示す。また、R1とR2、R3とR4はそれぞれ互いに結合して環を形成していてもよい。 The compound of the present invention is represented by the formula (1). In formula (1), R 1 to R 4 are the same or different and are a hydrogen atom, a halogen atom, a hydrocarbon group which may have a substituent, or a heterocyclic group which may have a substituent. , Carboxyl group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercapto group, substituted thio group, or substituted Or an unsubstituted amino group is shown. R 1 and R 2 , R 3 and R 4 may be bonded to each other to form a ring.
前記ハロゲン原子としては、フッ素、塩素、臭素及びヨウ素原子が挙げられる。炭化水素基としては、脂肪族炭化水素基、脂環式炭化水素基、芳香族炭化水素基、これらが複数結合した基が挙げられる。脂肪族炭化水素基として、例えば、メチル、エチル、プロピル、イソプロピル、ブチル、イソブチル、s−ブチル、t−ブチル、ヘキシル、デシル、ドデシル、テトラデシル、ヘキサデシル、ビニル、アリル、エチニル、1−プロピニル基などの炭素数1〜20(好ましくは1〜10、さらに好ましくは1〜8)程度の直鎖状又は分岐鎖状の脂肪族炭化水素基(アルキル基、アルケニル基、アルキニル基)などが挙げられる。脂環式炭化水素基としては、例えば、シクロプロピル、シクロブチル、シクロペンチル、シクロヘキシル、シクロヘキセニル、シクロオクチル、シクロデシル、シクロドデシル、ノルボルニル、アダマンチル、トリシクロ[5.2.1.02,6]デシル基などの炭素数3〜20(好ましくは3〜15)程度の脂環式炭化水素基(シクロアルキル基、シクロアルケニル基、橋架け炭素環式基等)などが挙げられる。芳香族炭化水素基としては、例えば、フェニル、ナフチル基などの炭素数6〜14程度の芳香族炭化水素基などが挙げられる。 Examples of the halogen atom include fluorine, chlorine, bromine and iodine atoms. Examples of the hydrocarbon group include an aliphatic hydrocarbon group, an alicyclic hydrocarbon group, an aromatic hydrocarbon group, and a group in which a plurality of these are bonded. Examples of the aliphatic hydrocarbon group include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, s-butyl, t-butyl, hexyl, decyl, dodecyl, tetradecyl, hexadecyl, vinyl, allyl, ethynyl, 1-propynyl group, and the like. And a linear or branched aliphatic hydrocarbon group (alkyl group, alkenyl group, alkynyl group) having about 1 to 20 carbon atoms (preferably 1 to 10, more preferably 1 to 8). Examples of the alicyclic hydrocarbon group include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, cyclooctyl, cyclodecyl, cyclododecyl, norbornyl, adamantyl, and tricyclo [5.2.1.0 2,6 ] decyl group. And an alicyclic hydrocarbon group having about 3 to 20 carbon atoms (preferably 3 to 15 carbon atoms) such as a cycloalkyl group, a cycloalkenyl group, and a bridged carbocyclic group. Examples of the aromatic hydrocarbon group include aromatic hydrocarbon groups having about 6 to 14 carbon atoms such as phenyl and naphthyl groups.
脂肪族炭化水素基と脂環式炭化水素基とが結合した基として、例えば、シクロペンチルメチル、シクロヘキシルメチル、シクロヘキシルエチル、4−メチルシクロヘキシル基などが挙げられる。また、脂肪族炭化水素基と芳香族炭化水素基とが結合した基として、例えば、ベンジル、2−フェニルエチル、1−フェニルエチル、3−フェニルプロピル等のアラルキル基;2−メチルフェニル、3−メチルフェニル、4−メチルフェニル基などが挙げられる。 Examples of the group in which an aliphatic hydrocarbon group and an alicyclic hydrocarbon group are bonded include cyclopentylmethyl, cyclohexylmethyl, cyclohexylethyl, 4-methylcyclohexyl group, and the like. Examples of the group in which an aliphatic hydrocarbon group and an aromatic hydrocarbon group are bonded include, for example, an aralkyl group such as benzyl, 2-phenylethyl, 1-phenylethyl, 3-phenylpropyl; 2-methylphenyl, 3-phenyl Examples include methylphenyl and 4-methylphenyl groups.
これらの炭化水素基は1又は2以上の置換基を有していてもよい。前記置換基として、例えば、ハロゲン原子(フッ素、塩素、臭素、ヨウ素原子)、複素環式基(ピロリジノ基、ピロリジノ基、モルホリノ基、イミダゾイル基、インドール基などの窒素原子、酸素原子及び硫黄原子から選択された少なくとも1種のヘテロ原子を含む3〜20員程度の複素環式基など)、カルボキシル基、置換オキシカルボニル基(アルコキシ−カルボニル基、シクロアルキルオキシカルボニル基、アリールオキシカルボニル基、アラルキルオキシカルボニル基など)、置換若しくは無置換カルバモイル基(カルバモイル基;メチルカルバモイル、ジメチルカルバモイル基等のモノ又はジ−炭化水素基置換カルバモイル基など)、シアノ基、アシル基(ホルミル、アセチル、プロピオニル基等の脂肪族アシル基;シクロヘキサンカルボニル基等の脂環式アシル基;ベンゾイル基等の芳香族アシル基;アセトアセチル基など)、ニトロ基、硫黄酸基(スルホン酸基、スルフィン酸基)、硫黄酸エステル基(スルホン酸エステル基、スルフィン酸エステル基)、ヒドロキシル基、置換オキシ基(アルコキシ基、シクロアルキルオキシ基、アリールオキシ基、アシルオキシ基など)、メルカプト基、置換チオ基(メチルチオ、エチルチオ基等のC1〜C6アルキルチオ基;シクロヘキシルチオ基等のシクロアルキルチオ基;フェニルチオ基等のアリールチオ基;アセチルチオ基等のアシルチオ基など)、置換若しくは無置換アミノ基(アミノ基、モノ又はジC1〜C6アルキルアミノ基など)、オキソ基これらが複数結合した基などが挙げられる。前記カルボキシル基、硫黄酸基、ヒドロキシル基、メルカプト基は保護基で保護されていてもよい。保護基としては、有機合成の分野で慣用の保護基を使用できる。 These hydrocarbon groups may have one or two or more substituents. Examples of the substituent include a halogen atom (fluorine, chlorine, bromine, iodine atom), a heterocyclic group (pyrrolidino group, pyrrolidino group, morpholino group, imidazolyl group, indole group, nitrogen atom, oxygen atom and sulfur atom). 3 to 20-membered heterocyclic group containing at least one selected hetero atom), carboxyl group, substituted oxycarbonyl group (alkoxy-carbonyl group, cycloalkyloxycarbonyl group, aryloxycarbonyl group, aralkyloxy) Carbonyl group etc.), substituted or unsubstituted carbamoyl group (carbamoyl group; mono- or di-hydrocarbon group substituted carbamoyl group such as methylcarbamoyl, dimethylcarbamoyl group etc.), cyano group, acyl group (formyl, acetyl, propionyl group etc.) Aliphatic acyl group; cyclohexa Alicyclic acyl groups such as carbonyl groups; aromatic acyl groups such as benzoyl groups; acetoacetyl groups, etc., nitro groups, sulfur acid groups (sulfonic acid groups, sulfinic acid groups), sulfur acid ester groups (sulfonic acid ester groups) , Sulfinic acid ester groups), hydroxyl groups, substituted oxy groups (alkoxy groups, cycloalkyloxy groups, aryloxy groups, acyloxy groups, etc.), mercapto groups, substituted thio groups (methylthio, ethylthio groups, etc.) C 1 -C 6 alkylthio A cycloalkylthio group such as a cyclohexylthio group; an arylthio group such as a phenylthio group; an acylthio group such as an acetylthio group), a substituted or unsubstituted amino group (amino group, mono- or di-C 1 -C 6 alkylamino group, etc.) , An oxo group or a group in which a plurality of these are bonded. The carboxyl group, sulfur acid group, hydroxyl group, and mercapto group may be protected with a protecting group. As the protecting group, a protecting group conventionally used in the field of organic synthesis can be used.
R1〜R4における複素環式基を構成する複素環には、芳香族性複素環及び非芳香族性複素環が含まれる。このような複素環としては、例えば、ヘテロ原子として酸素原子を含む複素環(例えば、フラン、テトラヒドロフラン、オキサゾール、イソオキサゾールなどの5員環、4−オキソ−4H−ピラン、テトラヒドロピラン、モルホリンなどの6員環、ベンゾフラン、イソベンゾフラン、4−オキソ−4H−クロメン、クロマン、イソクロマンなどの縮合環など)、ヘテロ原子としてイオウ原子を含む複素環(例えば、チオフェン、チアゾール、イソチアゾール、チアジアゾールなどの5員環、4−オキソ−4H−チオピランなどの6員環、ベンゾチオフェンなどの縮合環など)、ヘテロ原子として窒素原子を含む複素環(例えば、ピロール、ピロリジン、ピラゾール、イミダゾール、トリアゾールなどの5員環、ピリジン、ピリダジン、ピリミジン、ピラジン、ピペリジン、ピペラジンなどの6員環、インドール、インドリン、キノリン、アクリジン、ナフチリジン、キナゾリン、プリンなどの縮合環など)などが挙げられる。これらの複素環式基は、置換基(例えば、前記炭化水素基が有していてもよい置換基と同様の基、及び上記に例示の炭化水素基)を有していてもよい。 The heterocyclic ring constituting the heterocyclic group in R 1 to R 4 includes an aromatic heterocyclic ring and a non-aromatic heterocyclic ring. As such a heterocyclic ring, for example, a heterocyclic ring containing an oxygen atom as a hetero atom (for example, 5-membered ring such as furan, tetrahydrofuran, oxazole, isoxazole, 4-oxo-4H-pyran, tetrahydropyran, morpholine, etc. 6-membered ring, condensed ring such as benzofuran, isobenzofuran, 4-oxo-4H-chromene, chromane, isochroman, etc.), heterocycle containing a sulfur atom as a hetero atom (for example, 5 such as thiophene, thiazole, isothiazole, thiadiazole) 5-membered rings, 6-membered rings such as 4-oxo-4H-thiopyran, condensed rings such as benzothiophene, etc.), heterocycles containing nitrogen atoms as heteroatoms (eg, pyrrole, pyrrolidine, pyrazole, imidazole, triazole, etc.) Ring, pyridine, pyridazine, pyrimi Emissions, pyrazine, piperidine, 6-membered ring such as piperazine, indole, indoline, quinoline, acridine, naphthyridine, quinazoline, other condensed rings purine) and the like. These heterocyclic groups may have a substituent (for example, the same group as the substituent that the hydrocarbon group may have, and the hydrocarbon group exemplified above).
R1〜R4における置換オキシカルボニル基としては、例えば、メトキシカルボニル、エトキシカルボニル等のC1〜C6アルコキシ−カルボニル基;シクロヘキシルオキシカルボニル基等のシクロアルキルオキシカルボニル基;フェノキシカルボニル基等のアリールオキシカルボニル基;ベンジルオキシカルボニル基等のアラルキルオキシカルボニル基などが挙げられる。置換若しくは無置換カルバモイル基としては、例えば、カルバモイル基;メチルカルバモイル、ジメチルカルバモイル基等のモノ又はジ−炭化水素基置換カルバモイル基などが挙げられる。アシル基としては、例えば、ホルミル、アセチル、プロピオニル基等の脂肪族アシル基(C1〜C7脂肪族アシル基等);シクロヘキサンカルボニル基等の脂環式アシル基;ベンゾイル基等の芳香族アシル基;アセトアセチル基などが挙げられる。硫黄酸基として、スルホン酸基、スルフィン酸基が挙げられる。硫黄酸エステル基として、スルホン酸エステル基(スルホン酸C1〜C4アルキルエステル基等)、スルフィン酸エステル基(スルフィン酸C1〜C4アルキルエステル基等)が挙げられる。置換オキシ基としては、例えば、メトキシ、エトキシ、イソプロピルオキシ、ブトキシ基等のC1〜C6アルコキシ基;シクロヘキシルオキシ基等のシクロアルキルオキシ基;フェノキシ基等のアリールオキシ基;アセチルオキシ、プロピオニルオキシ基等のアシルオキシ基(C1〜C7アシルオキシ基等)などが挙げられる。置換チオ基としては、例えば、メチルチオ、エチルチオ基等のC1〜C6アルキルチオ基;シクロヘキシルチオ基等のシクロアルキルチオ基;フェニルチオ基等のアリールチオ基;アセチルチオ基等のアシルチオ基(C1〜C7アシルチオ基等)などが挙げられる。また、置換若しくは無置換アミノ基としては、例えば、アミノ基;メチルアミノ、ジメチルアミノ、エチルアミノ、ジエチルアミノ基等のモノ又はジC1〜C6アルキルアミノ基;アミジノ基;グアニジノ基等が例示される。 Examples of the substituted oxycarbonyl group in R 1 to R 4 include C 1 to C 6 alkoxy-carbonyl groups such as methoxycarbonyl and ethoxycarbonyl; cycloalkyloxycarbonyl groups such as cyclohexyloxycarbonyl group; and aryls such as phenoxycarbonyl group An oxycarbonyl group; and an aralkyloxycarbonyl group such as a benzyloxycarbonyl group. Examples of the substituted or unsubstituted carbamoyl group include a carbamoyl group; a mono- or di-hydrocarbon group-substituted carbamoyl group such as a methylcarbamoyl group and a dimethylcarbamoyl group. Examples of the acyl group include aliphatic acyl groups such as formyl, acetyl, propionyl groups (C 1 to C 7 aliphatic acyl groups); cycloaliphatic acyl groups such as cyclohexanecarbonyl groups; aromatic acyls such as benzoyl groups Group; acetoacetyl group and the like. Examples of the sulfur acid group include a sulfonic acid group and a sulfinic acid group. Examples of the sulfur acid ester group include a sulfonic acid ester group (such as a sulfonic acid C 1 to C 4 alkyl ester group) and a sulfinic acid ester group (such as a sulfinic acid C 1 to C 4 alkyl ester group). The substituted oxy group, for example, methoxy, ethoxy, isopropyloxy, C 1 -C 6 alkoxy group or a butoxy group; such as phenoxy group an aryloxy group; a cycloalkyl group such as cyclohexyl group, acetyloxy, propionyloxy And acyloxy groups such as C 1 -C 7 acyloxy groups. The substituted thio group, for example, methylthio, C 1 -C 6 alkylthio groups such as ethylthio group; arylthio group phenylthio group; cycloalkylthio groups such as cyclohexyl thio group acetylthio acylthio group such as (C 1 -C 7 Acylthio group, etc.). Examples of the substituted or unsubstituted amino group include amino groups; mono- or di-C 1 -C 6 alkylamino groups such as methylamino, dimethylamino, ethylamino and diethylamino groups; amidino groups; guanidino groups and the like. The
式(1)において、R1とR2、R3とR4が、それぞれ互いに結合して形成してもよい。環としては、例えば、シクロプロパン環、シクロブタン環、シクロペンタン環、シクロペンテン環、シクロヘキサン環、シクロヘキセン環、ノルボルナン環、ノルボルネン環、アダマンタン環などの3〜15員(好ましくは4〜12員、さらに好ましくは5〜8員)程度の非芳香族性炭素環(シクロアルカン環、シクロアルケン環、橋かけ炭素環)などが挙げられる。これらの環は、置換基(例えば、前記炭化水素基が有していてもよい置換基と同様の基)を有していてもよく、また他の環(非芳香族性環又は芳香族性環)が縮合していてもよい。 In the formula (1), R 1 and R 2 , R 3 and R 4 may be bonded to each other. Examples of the ring include 3 to 15 members (preferably 4 to 12 members, more preferably a cyclopropane ring, cyclobutane ring, cyclopentane ring, cyclopentene ring, cyclohexane ring, cyclohexene ring, norbornane ring, norbornene ring, and adamantane ring. Is a 5- to 8-membered) non-aromatic carbocycle (cycloalkane ring, cycloalkene ring, bridged carbocycle) and the like. These rings may have a substituent (for example, the same group as the substituent which the hydrocarbon group may have), and other rings (non-aromatic ring or aromaticity). Ring) may be condensed.
本発明においては、R1〜R4の少なくとも一つは水素原子以外の基である。具体的には、R1〜R4のうち2つが水素原子で2つが水素原子以外の基、又はR1〜R4のうち3つが水素原子で1つが水素原子以外の基、R1〜R4のすべてが水素原子以外の基である。特に、R1〜R4のうち3つが水素原子で1つが水素原子以外の基である化合物が好ましく用いられる。水素原子以外の基として、好ましくは、グリシン以外のアミノ酸の側鎖部位及びその誘導体が好ましく用いられる。ここで、アミノ酸の側鎖部位とは、アミノ酸のα炭素に結合する水素、アミノ基、カルボキシル基以外の官能基を意味しており、例えばアラニンにおけるメチル基、バリンにおけるイソプロピル基、アスパラギン酸におけるカルボキシメチル基、グルタミンにおけるカルバモイルエチル基、フェニルアラニンにおけるベンジル基等が挙げられる。なかでも、C1〜C12脂肪族炭化水素基、肪族炭化水素基と芳香族炭化水素基とが結合した基、及びこれらの誘導体、特にメチル基、及びベンジル基等が好ましい。 In the present invention, at least one of R 1 to R 4 is a group other than a hydrogen atom. Specifically, two of R 1 to R 4 are hydrogen atoms and two are groups other than hydrogen atoms, or three of R 1 to R 4 are hydrogen atoms and one is a group other than hydrogen atoms, R 1 to R All 4 are groups other than a hydrogen atom. In particular, a compound in which three of R 1 to R 4 are hydrogen atoms and one is a group other than a hydrogen atom is preferably used. As a group other than a hydrogen atom, a side chain site of an amino acid other than glycine and derivatives thereof are preferably used. Here, the side chain site of an amino acid means a functional group other than hydrogen, amino group, and carboxyl group that binds to the α-carbon of the amino acid, such as methyl group in alanine, isopropyl group in valine, carboxy group in aspartic acid Examples include a methyl group, a carbamoylethyl group in glutamine, a benzyl group in phenylalanine, and the like. Of these, a C 1 to C 12 aliphatic hydrocarbon group, a group in which an aliphatic hydrocarbon group and an aromatic hydrocarbon group are bonded, and derivatives thereof, particularly a methyl group and a benzyl group are preferable.
式(1)におけるR5〜R12は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、及び置換若しくは無置換アミノ基を示し、これらは、上記に例示のものを用いることができる。なかでも、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、ヒドロキシル基、置換オキシ基、メルカプト基、又は置換チオ基等が好ましく、特に、水素原子、C1〜C4アルキル基、ヒドロキシル基、C1〜C4アルキルオキシ基等が好ましく用いられる。 R 5 to R 12 in Formula (1) are the same or different and are a hydrogen atom, a halogen atom, a hydrocarbon group which may have a substituent, a heterocyclic group which may have a substituent, Carboxyl group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercapto group, substituted thio group, and substituted or An unsubstituted amino group is shown, and those exemplified above can be used. Of these, a hydrogen atom, a halogen atom, an optionally substituted hydrocarbon group, a hydroxyl group, a substituted oxy group, a mercapto group, or a substituted thio group is preferable, and in particular, a hydrogen atom, C 1 to C 4. alkyl group, a hydroxyl group, C 1 -C 4 alkyl group are preferably used.
Xにおける連結基としては、例えば、メチレン、メチルメチレン、ジメチルメチレン、エチレン、プロピレン、トリメチレン基などの直鎖状又は分岐鎖状のアルキレン基(C1〜C6アルキレン基等)などの2価の脂肪族炭化水素基;1,2−シクロペンチレン、1,3−シクロペンチレン、1,2−シクロヘキシレン、1,3−シクロヘキシレン、1,4−シクロヘキシレン、シクロペンチリデン、シクロヘキシリデン基等の2価の脂環式炭化水素基;カルボニル基;酸素原子(エーテル結合;−O−);硫黄原子(チオエーテル結合;−S−);オキシカルボニル基(エステル結合;−COO−、環Z1、環Z3、環Z5、環Z7と結合する側は右側であっても左側であってもよい);アミノカルボニル基(アミド結合;−CONH−、隣接する基と結合する側は右側であっても左側であってもよい);及びこれらが複数個結合した基などが挙げられる。好ましい連結基として、C1〜C4アルキレン基、酸素原子(エーテル結合;−O−)、硫黄原子(チオエーテル結合;−S−)及びこれらが2以上結合した基等が挙げられる。
Examples of the linking group in X include divalent groups such as linear or branched alkylene groups such as methylene, methylmethylene, dimethylmethylene, ethylene, propylene, and trimethylene groups (such as C 1 to C 6 alkylene groups). Aliphatic hydrocarbon group; 1,2-cyclopentylene, 1,3-cyclopentylene, 1,2-cyclohexylene, 1,3-cyclohexylene, 1,4-cyclohexylene, cyclopentylidene, cyclohexylidene Divalent alicyclic hydrocarbon group such as a group; carbonyl group; oxygen atom (ether bond; -O-); sulfur atom (thioether bond; -S-); oxycarbonyl group (ester bond; -COO-, ring Z 1 , ring Z 3 ,
環Z1及び環Z2における単環又は多環の窒素原子含有環としては、環を構成する原子として少なくとも一つの窒素原子を含んでいれば特に限定されず、芳香族性複素環及び非芳香族性複素環のいずれであってもよい。このような環には、例えば、例えば、ピロール、ピロリジン、ピラゾール、イミダゾール、トリアゾールなどの窒素原子含有5員環、ピリジン、ピリダジン、ピリミジン、ピラジン、ピペリジン、ピペラジンなどの窒素原子含有6員環、インドール、インドリン、キノリン、アクリジン、ナフチリジン、キナゾリン、プリンなどの縮合環;オキサゾリジン、モルホリンなどの環を構成する原子に窒素原子と酸素原子を含む環などが挙げられる。これらの環は、置換基を有していてもよい。置換基としては、上記と同様のものを利用でき、なかでも、アルキル基等の炭化水素基(C1〜C10の炭化水素基)、ヒドロキシル基、ハロゲン原子、ニトロ基、カルボキシ基、アルコキシカルボニル基、オキソ基、もしくは芳香族性複素環式基で置換されたアルキル基が挙げられる。このような置換基を有する環の代表的な例として、2−ピロリドン、スクシンイミド、マレイミド、プログルタミン酸などの窒素原子含有単環又は多環等が挙げられる。好ましくは、置換基を有していてもよい窒素原子含有5員環又は6員環が用いられ、より好ましくは、ピロリジン、ピペリジン、モルホリン、マレイミド、スクシンイミド等、特にピロリジン、ピペリジン等が用いられる。 The monocyclic or polycyclic nitrogen atom-containing ring in ring Z 1 and ring Z 2 is not particularly limited as long as it contains at least one nitrogen atom as an atom constituting the ring, and is an aromatic heterocyclic ring or non-aromatic ring. Any of the family heterocycles may be used. Such rings include, for example, nitrogen atom-containing 5-membered rings such as pyrrole, pyrrolidine, pyrazole, imidazole, and triazole, nitrogen-containing 6-membered rings such as pyridine, pyridazine, pyrimidine, pyrazine, piperidine, piperazine, and indole. And condensed rings such as indoline, quinoline, acridine, naphthyridine, quinazoline, and purine; and rings that contain a nitrogen atom and an oxygen atom in the atoms constituting the ring such as oxazolidine and morpholine. These rings may have a substituent. As the substituent, the same groups as described above can be used. Among them, a hydrocarbon group such as an alkyl group (C 1 to C 10 hydrocarbon group), a hydroxyl group, a halogen atom, a nitro group, a carboxy group, and an alkoxycarbonyl group. An alkyl group substituted with a group, an oxo group, or an aromatic heterocyclic group. Typical examples of the ring having such a substituent include nitrogen atom-containing monocyclic or polycyclic rings such as 2-pyrrolidone, succinimide, maleimide, and proglutamic acid. Preferably, a nitrogen atom-containing 5-membered ring or 6-membered ring which may have a substituent is used, more preferably pyrrolidine, piperidine, morpholine, maleimide, succinimide, etc., particularly pyrrolidine, piperidine and the like are used.
本発明の好ましい化合物としては、例えば、前記式(1)中、R1〜R4が、水素原子、又は置換基を有していてもよい炭化水素基であり、R5〜R12が、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、ヒドロキシル基、置換オキシ基、メルカプト基、又は置換チオ基であり、XがC1〜C4アルキレン基、酸素原子(エーテル結合;−O−)、又は硫黄原子(チオエーテル結合;−S−)であり、環Z1及び環Z2が、同一又は異なって、置換基を有していてもよい5員環又は6員環の窒素原子含有環であって、R1〜R4のうち少なくとも一つは置換基を有していてもよい炭化水素基である化合物が挙げられる。本発明の化合物として代表的な例として、前記式(2)で表される化合物を含む化合物の構造式を図2〜図9に示す。 As a preferable compound of the present invention, for example, in the formula (1), R 1 to R 4 are a hydrogen atom or a hydrocarbon group which may have a substituent, and R 5 to R 12 are same or different, a hydrogen atom, a halogen atom, or an optionally substituted hydrocarbon group, a hydroxyl group, a substituted oxy group, a mercapto group, or a substituted thio group, X is C 1 -C 4 alkylene group , An oxygen atom (ether bond; —O—), or a sulfur atom (thioether bond; —S—), and ring Z 1 and ring Z 2 may be the same or different and may have a substituent. Examples of the compound include a member ring or a six-membered nitrogen atom-containing ring, and at least one of R 1 to R 4 is a hydrocarbon group which may have a substituent. As typical examples of the compound of the present invention, structural formulas of compounds containing the compound represented by the formula (2) are shown in FIGS.
本発明の化合物は、公知の方法を用いて製造することができる。具体的には、例えば、式(1)におけるR1とR3の少なくとも一方及びR3とR4の少なくとも一方が水素原子以外の基である化合物は、次の工程に従って得ることができる。すなわち、下記式(3)
(式中、R5〜R10及びXは前記に同じ)
で表される化合物を、当該式(3)に含まれる一方のアミノ基を保護基(例えばtert−ブトキシカルボニル基:Boc等)で保護した後、下記式(3a)
(式中、Z1、R1、R2及びXは前記に同じ。但し、R1とR2の少なくとも一方が水素原子以外の基を示す)
で表される化合物と反応させ、次いで保護基で保護されたアミノ基を脱保護し、下記式(3b)
(式中、Z2、R3、R4は前記に同じ。但し、R3とR4の少なくとも一方が水素原子以外の基を示す)
で表される化合物と反応させる工程(A)により得ることができる。
The compound of this invention can be manufactured using a well-known method. Specifically, for example, a compound in which at least one of R 1 and R 3 and at least one of R 3 and R 4 in formula (1) is a group other than a hydrogen atom can be obtained according to the following step. That is, the following formula (3)
(Wherein R 5 to R 10 and X are the same as above)
After protecting one amino group contained in the formula (3) with a protecting group (for example, tert-butoxycarbonyl group: Boc etc.), the compound represented by the following formula (3a)
(Wherein Z 1 , R 1 , R 2 and X are the same as above, provided that at least one of R 1 and R 2 represents a group other than a hydrogen atom)
And then deprotecting the amino group protected with a protecting group, and the following formula (3b)
(In the formula, Z 2 , R 3 and R 4 are the same as above, provided that at least one of R 3 and R 4 represents a group other than a hydrogen atom.)
It can obtain by the process (A) made to react with the compound represented by these.
また、式(1)におけるR1とR2が共に水素原子である化合物は、例えば、次の工程に従って得ることができる。すなわち、前記式(3)で表される化合物を、当該式(3)に含まれる一方のアミノ基を保護基で保護した後、下記式(3c)
で表される化合物と反応させ、次いで下記式(4a)
(式中、環Z1は前記に同じ)
で表される化合物と反応させ、次いで保護基で保護されたアミノ基を脱保護した後、前記式(3b)で表される化合物を導入する工程(B)により得ることができる。
Moreover, the compound whose R < 1 > and R < 2 > in Formula (1) are both hydrogen atoms can be obtained according to the following process, for example. That is, after protecting one amino group contained in the formula (3) with a protecting group in the compound represented by the formula (3), the following formula (3c)
And then reacting with the compound represented by formula (4a)
(Wherein ring Z 1 is the same as above)
And then deprotecting the amino group protected by the protecting group, and then introducing the compound represented by the formula (3b) (B).
式(1)におけるR3とR4が共に水素原子である化合物は、例えば、上記工程(A)中、式(3c)で表される化合物に代えて前記式(3c)で表される化合物を用いて反応させ、次いで下記式(4b)
(式中、環Z2は前記に同じ)
で表される化合物を反応させる工程(C)により得ることができる。
The compound in which R 3 and R 4 in the formula (1) are both hydrogen atoms is, for example, a compound represented by the formula (3c) instead of the compound represented by the formula (3c) in the step (A). And then the following formula (4b)
(Wherein ring Z 2 is the same as above)
It can obtain by the process (C) with which the compound represented by this is made to react.
前記工程(A)〜(C)において、式(3a)〜(3c)及び式(4a)、(4b)で表される化合物の使用量は、式(2)で表される化合物1当量に対して、それぞれ例えば0.05〜50当量、好ましくは0.1〜10当量、さらに好ましくは0.3〜5当量程度である。式(3a)〜(3c)及び式(4a)、(4b)で表される化合物は、式(3)で表される化合物と同程度の当量で用いることもできる。 In said process (A)-(C), the usage-amount of the compound represented by Formula (3a)-(3c) and Formula (4a), (4b) is 1 equivalent of the compound represented by Formula (2). On the other hand, it is about 0.05 to 50 equivalents, preferably 0.1 to 10 equivalents, and more preferably about 0.3 to 5 equivalents. The compounds represented by the formulas (3a) to (3c) and the formulas (4a) and (4b) can be used in the same equivalent amount as the compound represented by the formula (3).
ここで、前記式(3a)及び(3b)で表される化合物は、例えば、原料としてアミノ酸を用い、当該アミノ酸のカルボキシル基を保護基で保護した後、α−アミノ基へ環Z1を導入し、次いで前記保護基を脱保護してカルボキシル基とする工程(D)に従って得ることができる。 Here, the compounds represented by the formulas (3a) and (3b) use, for example, an amino acid as a raw material, and after protecting the carboxyl group of the amino acid with a protecting group, introduce the ring Z 1 into the α-amino group Then, the protective group can be deprotected to give a carboxyl group (D).
前記工程(A)〜(D)における各反応は、溶媒の存在下又は非存在下で行われる。前記溶媒としては、例えば、ヘキサン、ヘプタン、オクタンなどの脂肪族炭化水素;シクロヘキサンなどの脂環式炭化水素;ベンゼン、トルエン、キシレン、エチルベンゼンなどの芳香族炭化水素;クロロホルム、ジクロロメタン、1,2−ジクロロエタンなどのハロゲン化炭化水素;ジエチルエーテル、ジメトキシエタン、テトラヒドロフラン、ジオキサンなどのエーテル;アセトン、メチルエチルケトンなどのケトン;酢酸メチル、酢酸エチル、酢酸イソプロピル、酢酸ブチルなどのエステル;N,N−ジメチルホルムアミド、N,N−ジメチルアセトアミドなどのアミド;アセトニトリル、プロピオニトリル、ベンゾニトリルなどのニトリルなどが挙げられる。これらの溶媒は単独で又は2種以上を混合して用いられる。 Each reaction in the steps (A) to (D) is performed in the presence or absence of a solvent. Examples of the solvent include aliphatic hydrocarbons such as hexane, heptane, and octane; alicyclic hydrocarbons such as cyclohexane; aromatic hydrocarbons such as benzene, toluene, xylene, and ethylbenzene; chloroform, dichloromethane, 1,2- Halogenated hydrocarbons such as dichloroethane; ethers such as diethyl ether, dimethoxyethane, tetrahydrofuran and dioxane; ketones such as acetone and methyl ethyl ketone; esters such as methyl acetate, ethyl acetate, isopropyl acetate and butyl acetate; N, N-dimethylformamide; Amides such as N, N-dimethylacetamide; nitriles such as acetonitrile, propionitrile, and benzonitrile. These solvents are used alone or in admixture of two or more.
各反応の温度は、基質の種類、反応の種類などに応じて、例えば0〜150℃、好ましくは20〜120℃、さらに好ましくは40〜80℃程度の範囲から選択でき、反応の種類によっては常温で行うことも可能である。各反応は常圧又は加圧下で行うことができる。反応時間は、反応温度及び圧力に応じて、例えば30分〜48時間程度の範囲から適当に選択できる。反応終了後、反応生成物は、例えば、液性調整、濾過、濃縮、晶析、洗浄、再結晶、抽出、蒸留、昇華精製、カラムクロマトグラフィー等の一般的な分離精製手段により分離精製できる。 The temperature of each reaction can be selected from the range of, for example, 0 to 150 ° C., preferably 20 to 120 ° C., more preferably 40 to 80 ° C., depending on the type of substrate, the type of reaction, etc. It is also possible to carry out at room temperature. Each reaction can be performed at normal pressure or under pressure. The reaction time can be appropriately selected from the range of about 30 minutes to 48 hours, for example, depending on the reaction temperature and pressure. After completion of the reaction, the reaction product can be separated and purified by general separation and purification means such as liquid property adjustment, filtration, concentration, crystallization, washing, recrystallization, extraction, distillation, sublimation purification, column chromatography and the like.
本発明の化合物は、正常型プリオンタンパク質に強固に結合して、構造変換を阻止する作用がある。このため、本発明の化合物は、正常型プリオンタンパク質の構造変換抑制剤やプリオン病の予防・治療剤の有効成分として有用である。本発明の化合物は、また、食品や飲料に添加して利用することもできる。ここで、本発明におけるプリオン病とは、正常型プリオンタンパク質が構造変換して生成される感染型プリオンタンパク質により引き起こされる疾患であって、例えば、羊のスクレイピー、ウシ海綿状脳症、クロイツフェルト・ヤコブ病、GSS、FFI、クールーおよび変異型ヤコブ病等が含まれる。 The compound of the present invention has an action of binding to a normal prion protein and preventing structural transformation. Therefore, the compound of the present invention is useful as an active ingredient of a structure conversion inhibitor of normal prion protein and a prophylactic / therapeutic agent for prion disease. The compound of the present invention can also be used by adding to foods and beverages. Here, the prion disease in the present invention is a disease caused by an infectious prion protein produced by structural conversion of a normal prion protein, such as sheep scrapie, bovine spongiform encephalopathy, Creutzfeldt-Jakob. Diseases, GSS, FFI, Kuru, mutant Jacob disease and the like.
本発明のプリオンタンパク質構造変換抑制剤及びプリオン病の予防・治療剤は、上記本発明の化合物を有効成分として含み、さらに、薬学的に許容される他の成分を含んでいてもよい。これらの薬剤は、ヒト、ウシ、ヒツジ、ウマ等の哺乳類などの動物に利用できる。前記薬学的に許容される成分としては、例えば、主に固形剤に用いられる賦形剤、滑沢剤、結合剤、崩壊剤、主に液剤に用いられる溶剤、溶解補助剤、懸濁化剤、等張化剤、緩衝剤、無痛化剤などが挙げられ、さらに防腐剤、抗酸化剤、着色剤、甘味剤、吸着剤、湿潤剤などを用いることもできる。 The prion protein structure conversion inhibitor and the prion disease preventive / therapeutic agent of the present invention contain the compound of the present invention as an active ingredient, and may further contain other pharmaceutically acceptable ingredients. These drugs can be used for animals such as mammals such as humans, cows, sheep and horses. Examples of the pharmaceutically acceptable ingredients include excipients, lubricants, binders, disintegrants mainly used for solid preparations, solvents, dissolution aids, and suspending agents mainly used for liquid preparations. , Tonicity agents, buffering agents, soothing agents, and the like, and antiseptics, antioxidants, coloring agents, sweetening agents, adsorbents, wetting agents, and the like can also be used.
剤形としては、例えば、水、生理食塩水等の希釈液又は分散媒に有効量の化合物を溶解、分散、乳化させた注射剤、クリーム、軟膏、飲料剤、エアロゾル、皮膚ゲル、点眼剤、点鼻剤等の液剤;錠剤、カプセル剤、散剤、顆粒剤、錠剤、除法剤、坐薬等の固形剤などを用いることができ、これらは有効成分がリポソームや徐放性材料等に封入された封入体や担体に担持された担持体などの形態であってもよい。 Examples of the dosage form include injections, creams, ointments, beverages, aerosols, skin gels, eye drops, in which an effective amount of a compound is dissolved, dispersed, and emulsified in a diluent or dispersion medium such as water or physiological saline. Liquid preparations such as nasal drops; solid preparations such as tablets, capsules, powders, granules, tablets, remedies, suppositories, etc. can be used, and these active ingredients are encapsulated in liposomes, sustained-release materials, etc. It may be in the form of an enclosure or a carrier carried on a carrier.
薬剤の投与は、経口、非経口のいずれであってもよく、動脈内、静脈内、筋肉内、皮下、腹腔内、直腸内へ、又は経呼吸、経皮、経鼻、経眼等による全身又は局所への投与等の方法により行うことができる。また、本発明の薬剤は、脳室内ポンプによって脳へ直接的に投与することもできる。 The drug can be administered either orally or parenterally, and it can be administered into the whole body by intraarterial, intravenous, intramuscular, subcutaneous, intraperitoneal, intrarectal or by respiration, percutaneous, nasal, ophthalmic, etc. Or it can carry out by methods, such as local administration. Moreover, the chemical | medical agent of this invention can also be directly administered to a brain with an intraventricular pump.
本発明のプリオンタンパク質構造変換抑制剤及びプリオン病の予防・治療剤の投与量は、それぞれ有効成分である化合物の種類や投与方法、さらに投与対象の種、年齢、体重、症状、薬物特性、病歴などに応じて異なるが、例えば、1回あたり体重1kg当たり数μgから数十mgの範囲であり、毎週数回から1日数回投与することができる。 The doses of the prion protein structure conversion inhibitor and the prion disease preventive / therapeutic agent of the present invention are the types of the active compound, the administration method, the species to be administered, age, weight, symptoms, drug characteristics, medical history, respectively. For example, the dose is in the range of several μg to several tens of mg per kg of body weight per time, and can be administered several times a week to several times a day.
薬理活性試験は、正常型プリオンタンパク質の構造変換抑制活性を評価できる方法であれば特に限定されないが、例えば、被検物質の存在下、プリオン感染細胞が生成する感染型プリオンタンパク質の生成を検出する手段を用いることができる。前記プリオン感染細胞は、プリオンに感染しうる細胞に、公知の方法でプリオンを感染させることにより生成できる。感染型プリオンタンパク質の検出は、特定のタンパク質を検出可能な公知の方法を用いることができ、好ましくは定量的な検出方法が用いられる。前記定量的な検出方法としては、例えば抗体、核酸、これらの類似体(ペプチド、PNAなど)等の特定のタンパク質を認識する手段と、蛍光体や放射線等で標識したタンパク質のイメージ解析手段(ELISA、ECL-plusウェスタンブロッティング等)等の認識されたタンパク質を定量化する手段とを組み合わせて行われる場合が多い。 The pharmacological activity test is not particularly limited as long as it is a method capable of evaluating the structure conversion inhibitory activity of normal prion protein. For example, the detection of infectious prion protein produced by prion-infected cells in the presence of a test substance is detected. Means can be used. The prion-infected cell can be generated by infecting a prion-infected cell with a prion by a known method. For detection of infectious prion protein, a known method capable of detecting a specific protein can be used, and a quantitative detection method is preferably used. As the quantitative detection method, for example, a means for recognizing a specific protein such as an antibody, a nucleic acid, or an analog thereof (peptide, PNA, etc.), and an image analysis means (ELISA) for a protein labeled with a phosphor or radiation. , ECL-plus Western blotting, etc.) are often combined with a means for quantifying a recognized protein.
薬理活性試験の具体例としては、マウス神経細胞株にプリオンを感染させ、次いで種々の濃度で被検物質(薬剤)を添加した培地で一定期間培養した後、タンパク質を回収して感染型プリオンタンパク質を抗体等で検出し、イメージ解析等により定量化する方法等を利用できる。上記試験において、感染型プリオンタンパク質の生成量が少ないほど、プリオンタンパク質の構造変化抑制活性に優れ、プリオン病の治療・予防に高い効果を奏すると評価できる。 As a specific example of a pharmacological activity test, a mouse neuron cell line is infected with prion, and then cultured for a certain period in a medium to which a test substance (drug) is added at various concentrations. And the like can be used for the detection of antibodies with antibodies and the like and quantification by image analysis or the like. In the above test, it can be evaluated that the smaller the amount of infectious prion protein produced, the better the prion protein structural change inhibitory activity and the higher the effect of treating and preventing prion diseases.
上記試験方法において、本発明の化合物は、同化合物を添加しないときの感染型プリオンタンパク質の生成量を100としたとき、同生成量を相対値として、例えば20以下、好ましくは15以下、より好ましくは13.5以下、特に12以下程度まで低減する効果を奏する。本発明の化合物は、また、本発明者らによってすでに報告されている2−ピロリジン−1−イル−N−[4−[4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル]−フェニル]−アセトアミド(特開2005−120002号公報参照)と比較して、感染型プリオンタンパク質の生成を抑制する効果に優れ、特に、前記式(2)で表される化合物により高い効果を得ることができる。このように、本発明の化合物は、正常型プリオンタンパク質の構造変換抑制効果に極めて優れるため、プリオンタンパク質構造変換抑制剤やプリオン病の予防・治療剤の有効成分として用いることにより、プリオン病の発症を効果的に予防し、発症後には症状の進行を阻止又は遅延することにより優れた治療効果を発揮することができる。 In the above test method, the compound of the present invention has a relative value of, for example, 20 or less, preferably 15 or less, and more preferably 15 or less, when the production amount of infectious prion protein when the compound is not added is 100. Has an effect of reducing it to 13.5 or less, particularly about 12 or less. The compounds of the present invention are also represented by 2-pyrrolidin-1-yl-N- [4- [4- (2-pyrrolidin-1-yl-acetylamino) -benzyl]-which has already been reported by the inventors. Compared with phenyl] -acetamide (see Japanese Patent Application Laid-Open No. 2005-120002), it has an excellent effect of suppressing the production of infectious prion protein, and in particular, a high effect is obtained with the compound represented by the formula (2). Can do. Thus, since the compound of the present invention is extremely excellent in the effect of suppressing the structural conversion of normal prion protein, it can be used as an active ingredient of a prion protein structural conversion inhibitor or a preventive / therapeutic agent for prion disease. Can be effectively prevented, and after the onset of symptoms, an excellent therapeutic effect can be exhibited by preventing or delaying the progression of symptoms.
本発明のキットは、化合物及び/又はその誘導体と、プリオンに感染した細胞と、感染型プリオンタンパク質を検出する手段とを含んでいる。プリオンに感染した細胞、及び感染型プリオンタンパク質を検出する手段としては、上記薬理評価試験において例示のものを用いることができる。本発明のキットは、プリオンタンパク質の構造変換を抑制する化合物のスクリーニングや、同化合物を用いた試験、評価、研究等に利用できる。 The kit of the present invention includes a compound and / or a derivative thereof, a cell infected with a prion, and a means for detecting an infectious prion protein. As a means for detecting cells infected with prions and infectious prion protein, those exemplified in the pharmacological evaluation test can be used. The kit of the present invention can be used for screening for a compound that suppresses the structural conversion of prion protein, testing, evaluation, research, and the like using the compound.
本発明の化合物は、また、当該化合物をベースとして、上記医薬用途により最適化された化合物を効率よくスクリーニングする方法に利用することができる。スクリーニングは、in vivo、in vitro、in silico等による公知の手法を利用できる。特に本発明では、in silicoスクリーニング(バーチャルスクリーニング)で得られた候補化合物について、in vitro及び/又はin vivoスクリーニングを行う方法を用いると、優れた薬理効果を発揮しうる化合物を効率よく選別でき好ましい。前記in silicoスクリーニグには、例えば、後述のドッキングシミュレーションを用いたスクリーニング法が好ましく用いられる。 The compound of the present invention can also be used in a method for efficiently screening a compound optimized for the above pharmaceutical use based on the compound. For screening, a known technique such as in vivo, in vitro, in silico or the like can be used. In particular, in the present invention, it is preferable to use a method of performing in vitro and / or in vivo screening for a candidate compound obtained by in silico screening (virtual screening), because it is possible to efficiently select a compound that can exert an excellent pharmacological effect. . For the in silico screening, for example, a screening method using docking simulation described later is preferably used.
本発明の方法は、上記本発明の化合物及び/又はその誘導体を用い、正常型プリオンタンパク質とのドッキングシミュレーションにより予測される結合強さに基づき、プリオンタンパク質の構造変換抑制活性を有する化合物をスクリーニングする方法である。スクリーニングには、通常、ライブラリが用いられるが、当該ライブラリは、本発明の誘導体で構成されていても良く、ソフトウェア等を用いてコンピューター上で作成された公知乃至未知の化合物であってもよく、市販のものを利用することもできる The method of the present invention uses the compound of the present invention and / or a derivative thereof to screen for a compound having a prion protein structure conversion inhibitory activity based on the binding strength predicted by docking simulation with a normal prion protein. Is the method. For screening, a library is usually used. The library may be composed of the derivative of the present invention, and may be a known or unknown compound created on a computer using software or the like. Commercially available products can also be used
前記本発明の化合物の誘導体には、例えば、本発明の化合物に公知の置換基を導入等することにより部分的に改変された化合物等が含まれる。このような置換基としては、例えば上記に例示のものを利用できる。但し、本発明の化合物の誘導体には、2−ピロリジン−1−イル−N−[4−[4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル]−フェニル]−アセトアミドは含まれない。上記本発明の化合物の誘導体でライブラリを構成して、本発明の化合物の構造を用いたリード化合物の最適化に利用することができる。以下、ドッキングシミュレーションに用いる本発明の化合物、その誘導体、及びライブラリを「被検物質」と総称する場合がある。 The derivative of the compound of the present invention includes, for example, a compound partially modified by introducing a known substituent into the compound of the present invention. As such substituents, for example, those exemplified above can be used. However, derivatives of the compounds of the present invention include 2-pyrrolidin-1-yl-N- [4- [4- (2-pyrrolidin-1-yl-acetylamino) -benzyl] -phenyl] -acetamide. Absent. A library can be constructed with the above-described derivatives of the compound of the present invention and used for optimizing lead compounds using the structure of the compound of the present invention. Hereinafter, the compounds of the present invention, derivatives thereof, and libraries used for docking simulation may be collectively referred to as “test substances”.
ドッキングシミュレーションは、一般に、被検物質及び正常型プリオンタンパク質の各立体構造情報に基づき、ドッキングシミュレーション用ソフトウェアプログラムによりコンピューター上で実施される。前記立体構造情報は、公知のタンパク質に関してはProtein Data Bank(PDB)などの公知のデータベースの登録情報として入手でき、新規物質に関しては汎用のプログラム等を用いてモデリングする等の方法により個別に作成することができる。 The docking simulation is generally performed on a computer by a docking simulation software program based on the three-dimensional structure information of the test substance and the normal prion protein. The three-dimensional structure information can be obtained as registered information in a known database such as Protein Data Bank (PDB) for known proteins, and individually created by a method such as modeling using a general-purpose program for new substances. be able to.
本発明の被検物質の立体構造情報は、例えばCambridgeSoft社製のChemOffice等のモデリングソフトウェアを用いて作成することができる。例えば、ChemOfficeを用いてGUI上の手動操作で個別にモデリングした後、ChemOfficeに付属の半経験的分子モデリングアプリケーションCSMOPACでAM1法を用いてエネルギーの最小化を行うことにより立体構造情報を作成することができる。こうして得られた情報に、さらに、電荷の付加や水素原子の削除などの処理を施すことにより、タンパク質との相互作用をより精度良く解析することができる。具体的には、MQEq法(修正電荷平衡法:Chem-Bio Info. J. 1, 35-40(2001)等)を用いて電荷を付加した。本発明の化合物及びライブラリ化合物の立体構造情報は、公知の方法で得ることができ、上記方法に限定されない。 The three-dimensional structure information of the test substance of the present invention can be created using modeling software such as ChemOffice manufactured by CambridgeSoft. For example, after modeling individually by manual operation on the GUI using ChemOffice, create 3D structure information by minimizing energy using the AM1 method with the semi-empirical molecular modeling application CSMOPAC attached to ChemOffice. Can do. By further processing the information obtained in this manner, such as addition of electric charge or deletion of hydrogen atoms, the interaction with the protein can be analyzed with higher accuracy. Specifically, charges were added using the MQEq method (modified charge equilibrium method: Chem-Bio Info. J. 1, 35-40 (2001), etc.). The three-dimensional structure information of the compound of the present invention and the library compound can be obtained by a known method, and is not limited to the above method.
プリオンタンパク質の立体構造情報は、Protein Data Bank(PDB)より入手可能であり、例えば、正常型マウスプリオンタンパク質の立体構造情報は、NMRにより構造決定された登録情報PDB code 1AG2を利用できる。さらに、電荷の付加や水素原子の削除などの処理を、AutoDockTools(Scripps Research Institute社製)を用い、Amber United-atom modelに従って行うことができる。 The three-dimensional structure information of prion protein can be obtained from Protein Data Bank (PDB). For example, the three-dimensional structure information of normal mouse prion protein can use registration information PDB code 1AG2 whose structure is determined by NMR. Furthermore, processes such as addition of charges and deletion of hydrogen atoms can be performed according to the Amber United-atom model using AutoDockTools (manufactured by Scripps Research Institute).
ドッキングシミュレーションによる結合強さの予測は、例えば、正常型プリオンタンパク質に存在する結合ポケットに焦点を当てて、被検物質と正常型プリオンタンパク質との結合自由エネルギーを予測し、当該エネルギーが低いほど両者の結合が強いと評価する方法により行うことができる。例えば、特開2005−120002号公報に開示される方法を利用できる。 Prediction of binding strength by docking simulation, for example, focuses on the binding pockets present in normal prion protein, predicts the binding free energy between the test substance and normal prion protein, It can be carried out by a method for evaluating that the bond is strong. For example, a method disclosed in JP 2005-120002 A can be used.
前記結合ポケットとしては、例えば、A-S2ループとヘリックスBのC端側との間にある結合ポケットを用いることができ、「結合ポケットに焦点を当てる」とは、当該結合サイトを中心に十分な大きさのグリッドボックスを設定することを意味している。グリッドボックスのサイズは特に限定されないが、例えば45Å×45Å×30Åのサイズに設定できる。 As the binding pocket, for example, a binding pocket between the A-S2 loop and the C-end side of helix B can be used, and “focusing on the binding pocket” means that the binding site is sufficiently centered. This means setting a grid box of a large size. The size of the grid box is not particularly limited, but can be set to, for example, a size of 45 mm × 45 mm × 30 mm.
ドッキングシミュレーション用ソフトウェアプログラムとしては、ドッキングシミュレーション用途の一又は複数のソフトウェアプログラムを組み合わせて利用することができ、例えば、DOCK、AutoDock、GOLDなどの公知のものを利用できる。例えば、前記AutoDockによれば、正常型プリオンタンパク質の位置を固定し、化合物の構造変化のみ考慮したフレキシブルドッキング法による自動ドッキングシミュレーションを行うことができる。 As the software program for docking simulation, one or a plurality of software programs for docking simulation can be used in combination, and for example, known programs such as DOCK, AutoDock, and GOLD can be used. For example, according to the AutoDock, it is possible to perform an automatic docking simulation by the flexible docking method in which the position of the normal prion protein is fixed and only the structural change of the compound is considered.
より詳細には、AutoDockを用いた自動ドッキングシミュレーションは、ライブラリと正常型プリオンタンパク質について、ラマルク的遺伝的アルゴリズムによって最適な結合状態を探索し、一つの化合物に対して、1回当たり数千万のドッキングモードを評価する方法を複数回繰り返し行い、最も低い自由エネルギーを当該化合物とプリオンタンパク質との結合自由エネルギーと予測する。一方、本発明の化合物と正常型プリオンタンパク質について、上記と同様の方法で結合自由エネルギーを予測する。こうして得られた結合自由エネルギーの値が低いものほど結合が強い、すなわち、プリオンタンパク質の構造変化を抑制する効果に優れていると評価することができる。 More specifically, the automatic docking simulation using AutoDock searches the optimal binding state of the library and normal prion protein by a Lamarckian genetic algorithm, and tens of millions per compound for each compound. The method for evaluating the docking mode is repeated several times, and the lowest free energy is predicted as the binding free energy between the compound and the prion protein. On the other hand, for the compound of the present invention and normal prion protein, the binding free energy is predicted by the same method as described above. It can be evaluated that the lower the value of the binding free energy thus obtained, the stronger the binding, that is, the better the effect of suppressing the structural change of the prion protein.
具体的には、前記式(2)で表される化合物は、上記方法に基づき予測される結合自由エネルギーが-11.43kcal/molと低く、また薬理活性試験による有効性が確認されている。この知見に基づき、結合自由エネルギーが-11.43kcal/mol未満の化合物、例えば図2〜図9に示される全ての化合物は、式(2)で表される化合物と同程度又はそれ以上の有用性があると推定される。 Specifically, the compound represented by the formula (2) has a binding free energy predicted based on the above method as low as −1.43 kcal / mol, and its effectiveness by a pharmacological activity test has been confirmed. Based on this finding, compounds having a binding free energy of less than −11.43 kcal / mol, for example, all the compounds shown in FIG. 2 to FIG. 9 are as useful as or more than the compounds represented by formula (2). It is estimated that there is.
式(1)で表される化合物中、結合自由エネルギーを指標としてプリオン病の治療・予防剤に有用と推定される化合物群は、例えば、結合自由エネルギーが−14.00kcal/mol未満の化合物(例えば、図2〜図9中の6LQ-C-5G、5LR-C-5G、MLR-C-5G、6LK-C-5G、5LA-S-6DH、6LN-C-5G、5G-C-6DH、6LR-C-5G、5LH-C-5G等)が挙げられ、次いで、同−14.00kcal/mol以上、−13.00kcal/mol未満の化合物群、同−13.00kcal/mol以上、−12.00kcal/mol未満の化合物群の順で有効であると推定できる。さらに、結合自由エネルギーが−12.00kcal/mol以上、−11.00kcal/mol未満の化合物群は、式(2)で表される化合物と同程度の結合自由エネルギーを有するため、式(2)の化合物と同程度の薬理活性を発揮する可能性が高いと推定できる。 Among the compounds represented by the formula (1), a compound group that is estimated to be useful as a therapeutic / preventive agent for prion diseases using binding free energy as an index includes, for example, compounds having binding free energy of less than −14.00 kcal / mol ( For example, 6LQ-C-5G, 5LR-C-5G, MLR-C-5G, 6LK-C-5G, 5LA-S-6DH, 6LN-C-5G, 5G-C-6DH in FIGS. , 6LR-C-5G, 5LH-C-5G, etc.), followed by a compound group of −14.00 kcal / mol or more and less than −13.00 kcal / mol, −13.00 kcal / mol or more, − It can be estimated that it is effective in the order of the compound group of less than 12.00 kcal / mol. Furthermore, since the compound group having a bond free energy of -12.00 kcal / mol or more and less than -11.00 kcal / mol has the same bond free energy as the compound represented by formula (2), the formula (2) It can be presumed that the possibility of exhibiting the same degree of pharmacological activity as that of the compound is high.
さらに、これらの化合物又はその誘導体を、プリオンタンパク質の構造変換抑制活性を有する化合物をスクリーニングする方法に用いて、正常型プリオンタンパク質とのドッキングシミュレーションにより予測される結合強さに基づき、プリオン病の治療・予防剤の有効成分の探索に利用することもできる。 Furthermore, these compounds or their derivatives are used in a method for screening a compound having a prion protein structure conversion inhibitory activity, and based on the binding strength predicted by docking simulation with a normal prion protein, treatment of prion disease -It can also be used to search for active ingredients of prophylactic agents.
本発明の化合物は、プリオンタンパク質と結合する公知の化合物と比較して、ドッキングシミュレーションによる正常型プリオンタンパク質との結合はより強く、しかも上記試験により優れた薬理活性を発揮することができる。そのため、本発明の化合物は、それ自体をプリオンタンパク質構造変換抑制剤やプリオン病の予防・治療剤の有効成分として利用できるだけでなく、当該化合物の誘導体からなるライブラリを作成することにより、さらなる薬理効果、生体適合性等に優れた薬剤の開発に好ましく利用できる。 Compared with known compounds that bind to prion protein, the compound of the present invention has stronger binding to normal prion protein by docking simulation and can exhibit superior pharmacological activity by the above test. Therefore, the compound of the present invention can be used not only as an active ingredient of a prion protein structure conversion inhibitor or a prion disease prophylactic / therapeutic agent itself, but also by creating a library comprising derivatives of the compound to further increase the pharmacological effect. It can be preferably used for the development of a drug excellent in biocompatibility and the like.
以下に、実施例に基づいて本発明をより詳細に説明するが、本発明はこれらの実施例により限定されるものではない。 Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.
実施例1
下記式(2)
で表される2−ピロリジン−1−イル−N−[4−{4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル}−フェニル]−プロピオンアミドの合成
4,4’−メチレンジアニリン12gに対し、1/2当量のジカルボン酸ジ−tert−ブチル(Boc無水物)を反応させ、アミンの片方のみがBoc基で保護された反応生成物について、ニンヒドリン試薬等を用いて、薄相クロマトグラフィー(TLC)で確認しながら順相カラムクロマトグラフィにより精製を行い、ほぼ当量の4−(4−tert−ブトキシカルボニルアミノベンジル)アニリン18gを得た。
Example 1
Following formula (2)
Synthesis of 2-pyrrolidin-1-yl-N- [4- {4- (2-pyrrolidin-1-yl-acetylamino) -benzyl} -phenyl] -propionamide represented by the formula 4,4′-methylenedi About 12 g of aniline, 1/2 equivalent of di-tert-butyl dicarboxylate (Boc anhydride) was reacted, and the reaction product in which only one of the amines was protected with a Boc group was diluted with a ninhydrin reagent or the like. Purification was performed by normal phase column chromatography while confirming by phase chromatography (TLC) to obtain 18 g of approximately equivalent 4- (4-tert-butoxycarbonylaminobenzyl) aniline.
得られた4−(4−tert−ブトキシカルボニルアミノベンジル)アニリン4gに対し、HBTU20.4gとHOBt8.2gをDMF100mL中に溶解した溶液を縮合剤に用いて、Fmoc-L-アラニンを反応させた。反応混合液を常温で1時間撹拌後、ジクロロメタンにより分配を行うことによって縮合剤を除去し、残渣を20%ピペリジン含有DMF溶液によりFmoc基を脱保護した。混合液を順相カラムクロマトグラフィにより精製を行い、下記式(X)
で表される化合物2.8gを得た。
Fmoc-L-alanine was reacted with 4 g of 4- (4-tert-butoxycarbonylaminobenzyl) aniline obtained using a solution obtained by dissolving 20.4 g of HBTU and 8.2 g of HOBt in 100 mL of DMF as a condensing agent. . After the reaction mixture was stirred at room temperature for 1 hour, the condensing agent was removed by partitioning with dichloromethane, and the Fmoc group was deprotected with a DMF solution containing 20% piperidine in the residue. The mixture is purified by normal phase column chromatography, and the following formula (X)
2.8 g of a compound represented by the formula:
得られた前記式(X)で表される化合物2gに対し、1,4−ジブロモブタン2.8gを、エタノール100mL中、NaHCO31gを共存させ、50℃で加熱撹拌することにより、下記式(Y)
で表される化合物を1.8g得た。得られた前記式(Y)で表される化合物1gに対し、ジクロロエタン中、TFAを反応させてBoc基を脱保護し、次いで以下に示す方法で1−ピロリジン酢酸との縮合反応を行った。DMF中、4当量の1−ピロリジン酢酸、HBTU、及びHOBtの共存下、室温で2時間撹拌後、溶媒を洗浄、留去し、次いで順相カラムクロマトグラフィ及び逆相HPLCにより精製を行い、目的の2−ピロリジン−1−イル−N−[4−{4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル}−フェニル]−プロピオンアミドを収率12.3%で得た。
以下、2−ピロリジン−1−イル−N−[4−{4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル}−フェニル]−プロピオンアミドを、「5G-C-5LA」と称する。
[目的化合物 のスペクトルデータ]
13C-NMR(CDCl3) δ:168.9, 137.0, 136.9, 136.1, 135.9, 129.4, 119.7, 119.7, 64.2, 59.8, 54.6, 51.2, 40.8, 38.6, 29.7, 24.1, 23.6, 16.4
HR-FAB-MS: m/z 434.2680 ([M+H]+)C26H34N4O2
By adding 2.8 g of 1,4-dibromobutane to 1 g of NaHCO 3 in 100 mL of ethanol and heating and stirring at 50 ° C. with respect to 2 g of the compound represented by the formula (X) thus obtained, the following formula (Y)
As a result, 1.8 g of a compound represented by the formula: 1 g of the obtained compound represented by the formula (Y) was reacted with TFA in dichloroethane to deprotect the Boc group, and then subjected to a condensation reaction with 1-pyrrolidineacetic acid by the method shown below. After stirring for 2 hours at room temperature in the presence of 4 equivalents of 1-pyrrolidineacetic acid, HBTU and HOBt in DMF, the solvent was washed and distilled off, and then purified by normal phase column chromatography and reverse phase HPLC. 2-Pyrrolidin-1-yl-N- [4- {4- (2-pyrrolidin-1-yl-acetylamino) -benzyl} -phenyl] -propionamide was obtained in a yield of 12.3%.
Hereinafter, 2-pyrrolidin-1-yl-N- [4- {4- (2-pyrrolidin-1-yl-acetylamino) -benzyl} -phenyl] -propionamide is referred to as “5G-C-5LA”. .
[Spectral data of target compound]
13 C-NMR (CDCl 3 ) δ: 168.9, 137.0, 136.9, 136.1, 135.9, 129.4, 119.7, 119.7, 64.2, 59.8, 54.6, 51.2, 40.8, 38.6, 29.7, 24.1, 23.6, 16.4
HR-FAB-MS: m / z 434.2680 ([M + H] + ) C 26 H 34 N 4 O 2
実施例2
実施例1で得た化合物5G-C-5LAを用いたin silicoスクリーニング
[ライブラリの作成]
実施例1で得た化合物に各種置換基を導入して得られる誘導体からなるライブラリを作成する。
[立体構造情報の入手]
実施例1で得た化合物の立体構造情報は、ChemOffice(CambridgeSoft社)を用いて、GUI操作によって手動で個別にモデリングした後に、ChemOfficeに付属のCS MOPACでAM1法を用いてエネルギー最小化を行い作成した。電荷付与は、中野らが開発したMQEq法(修正電荷平衡法)を用いた。
正常型プリオンタンパク質の立体構造情報は、Protein Data Bank(PDB)に登録されている、NMRによって構造決定された正常型マウスプリオン蛋白質(PDB code: 1AG2)を用いた。プリオン蛋白質の各原子に対する電荷情報付加およびその後の水素削除は、米国スクリプト研究所で開発され公開されているAutoDockToolsを用いて、Amber United-Atom model法で行った。
Example 2
In silico
A library composed of derivatives obtained by introducing various substituents into the compound obtained in Example 1 is prepared.
[Obtaining three-dimensional structure information]
The three-dimensional structure information of the compound obtained in Example 1 was individually modeled manually by GUI operation using ChemOffice (CambridgeSoft), and then energy minimization was performed using AM1 method with CS MOPAC attached to ChemOffice. Created. The charge was applied using the MQEq method (modified charge balance method) developed by Nakano et al.
For the three-dimensional structure information of the normal prion protein, a normal mouse prion protein (PDB code: 1AG2) registered in the Protein Data Bank (PDB) and whose structure was determined by NMR was used. Addition of charge information to each atom of the prion protein and subsequent hydrogen deletion were performed by the Amber United-Atom model method using AutoDockTools developed and published at the Script Institute of the United States.
[結合サイトの予測]
正常型プリオンタンパク質における実施例1で得た化合物との結合サイトをNMRの遅い揺らぎの情報に基づき予測した。こうして結合サイトと予測された付近を中心に、45Åx45Åx30Åの大きさのグリッドボックスを設定し、in silicoスクリーニングに利用した。
[in silicoスクリーニング]
ライブラリ化合物と正常型マウスプリオンタンパク質について、AutoDockを用いた自動ドッキングシミュレーションを行い、両者の結合自由エネルギーを予測した。具体的には、ラマルク的遺伝的アルゴリズムによって最適な結合状態を探索し、1つの化合物に対して、1回で2500万のドッキングモードを評価し、これを10回繰り返して、その中で最も低い自由エネルギーをその化合物とマウスプリオン蛋白質の結合自由エネルギーと予測した(J. Computational Chemistry, 19: 1639-1662 (1998))。上記シミュレーションで得られた実施例1で得た化合物の結合自由エネルギーは-11.43kcal/molであった。
[Coupling site prediction]
The binding site of the normal prion protein with the compound obtained in Example 1 was predicted based on information on the slow fluctuation of NMR. A grid box with a size of 45 mm x 45 mm x 30 mm was set around the vicinity predicted to be a binding site in this way and used for in silico screening.
[In silico screening]
For the library compound and normal mouse prion protein, an automatic docking simulation using AutoDock was performed to predict the binding free energy of both. Specifically, the optimal binding state is searched by a Lamarckian genetic algorithm, and 25 million docking modes are evaluated once for one compound, and this is repeated 10 times, and the lowest among them. The free energy was predicted as the binding free energy of the compound and mouse prion protein (J. Computational Chemistry, 19: 1639-1662 (1998)). The binding free energy of the compound obtained in Example 1 obtained by the above simulation was −11.43 kcal / mol.
上記シミュレーションの結果、ライブラリ化合物のなかでも、結合自由エネルギー値が実施例1で得た化合物より低かった化合物(-11.43kcal/mol未満)を表1に示す。表1中、「名称」は、それぞれ図2〜図9に表される構造式に対応する化合物を示す。 Table 1 shows compounds (less than −11.43 kcal / mol) whose binding free energy values were lower than those obtained in Example 1 among the library compounds as a result of the simulation. In Table 1, “name” indicates a compound corresponding to the structural formula shown in FIGS.
これらの結果から、表1に示される化合物は、実施例1で得た化合物5G-C-5LAと比較して、正常型プリオンタンパク質とより強固に結合することができ、当該結合を介して正常型プリオンタンパク質の構造変換を阻害する活性に優れる化合物であると容易に予測できる。ここで、5G-C-5LAは、後述の評価試験に示されるように、感染性プリオンタンパク質の生成を抑制するという優れた薬理活性を発揮する化合物であることが確認されている。従って、表1に示される化合物は、5G-C-5LAと同等又はそれ以上の薬理活性を発揮することができると予測することができる。従って、これらの化合物は、プリオン病の予防・治療剤を構成する有効成分として、又はそのような化合物を探索するリード化合物として極めて有用である。さらに、これらの化合物自体又はその誘導体を用いたin silicoスクリーニングにより、プリオンタンパク質の構造変換抑制活性を有する化合物を得ることにより、プリオン病の予防・治療に有用な化合物を容易に探索することができる。 From these results, the compounds shown in Table 1 can bind to the normal prion protein more firmly than the compound 5G-C-5LA obtained in Example 1, and normality is achieved through the binding. It can be easily predicted that the compound is excellent in activity to inhibit the structural conversion of the type prion protein. Here, 5G-C-5LA has been confirmed to be a compound that exhibits an excellent pharmacological activity of suppressing the production of infectious prion protein, as shown in an evaluation test described later. Therefore, it can be predicted that the compounds shown in Table 1 can exhibit pharmacological activity equivalent to or higher than that of 5G-C-5LA. Therefore, these compounds are extremely useful as an active ingredient constituting a preventive / therapeutic agent for prion diseases or as a lead compound for searching for such compounds. Further, by obtaining a compound having a structure conversion inhibitory activity of prion protein by in silico screening using these compounds themselves or derivatives thereof, compounds useful for the prevention / treatment of prion disease can be easily searched. .
評価試験
実施例1で得た化合物について以下の方法で薬理試験を行った。
[細胞]
マウス視床下部神経細胞系列GT1は、マウスプリオンに感染しうる。化合物の抗プリオン作用を評価する目的で、我々はGTFK-1細胞系列を用いた(N. Nishida et al., J. Virol, 74, 320-325 (2000))。これらはGSS由来の(O. Milhavet et al., Proc. Natl. Acad. Sci. U. S. A., 97, 13937-13942 (2000))マウス適合プリオンに安定に感染した、福岡-1株である。
[プリオン感染処理]
GT1-7細胞は、ペニシリン(GIBCO/BRL)を含むDMEM培地で培養した。3〜4日ごとに、細胞をトリプシン処理し、1:5に希釈して継代した。安定にFukuoka-1または22L株に感染したGT1-7細胞については、O.Milhavet et al., Proc. Natl. Acad. Sci. U.S.A. 97, 13937-13942 (2000)に記載されている。化合物のストック溶液(選択された59の化合物のうち19個)は、100mMのDMSO中に調製し4℃で保存した。使用前に、化合物を培地で希釈した。対照細胞を溶媒のみを含む(0.1%)培地で処理した。約2×105の細胞を6ウェルプレートの各ウェルに入れ、化合物を添加して15時間後に薬剤処理を開始した。72時間のインキュベーションの後に、細胞を回収した。
Evaluation test The compound obtained in Example 1 was subjected to a pharmacological test by the following method.
[cell]
Mouse hypothalamic neuronal lineage GT1 can infect mouse prions. In order to evaluate the anti-prion action of the compounds, we used the GT FK-1 cell line (N. Nishida et al., J. Virol, 74, 320-325 (2000)). These are Fukuoka-1 strains stably infected with mouse-compatible prions derived from GSS (O. Milhavet et al., Proc. Natl. Acad. Sci. USA, 97, 13937-13942 (2000)).
[Prion infection treatment]
GT1-7 cells were cultured in DMEM medium containing penicillin (GIBCO / BRL). Every 3-4 days, cells were trypsinized, diluted 1: 5 and passaged. GT1-7 cells stably infected with Fukuoka-1 or 22L strain are described in O. Milhavet et al., Proc. Natl. Acad. Sci. USA 97, 13937-13942 (2000). Compound stock solutions (19 of 59 selected compounds) were prepared in 100 mM DMSO and stored at 4 ° C. Prior to use, the compounds were diluted with media. Control cells were treated with medium containing only solvent (0.1%). Approximately 2 × 10 5 cells were placed in each well of a 6-well plate and drug treatment was initiated 15 hours after compound addition. Cells were harvested after 72 hours of incubation.
[感染型プリオンタンパク質の定量]
回収した細胞を、1×Triton/DOC溶解バッファー(0.5% tritonX-100, 0.5% deoxycholic acid, 150mM NaCl, 25mM Tris HCl pH7.5, 2mM EDTA およびペプスタチン、ロイペプシン)150μl中に溶解させた。タンパク質濃度をBCSアッセイ(Pierce社製)で測定し、各サンプルを2mgタンパク質に標準化した。PrPsc産生を分析するために、タンパク質を10μg/mgの濃度でプロテイナーゼKで30分間37℃で消化した。消化はPMSF(2mM)で阻害し、サンプルについて15% SDS/PAGEゲルを用いてSDS/PAGEを行った。次いで、タンパク質をPVDF膜(Immobilon-P, Amersham, USA社製)に転写した。PrPresのために、抗マウスPrP抗体(SS28)を用いた。またPrPCの検出のために、SAF32抗体(Spi-Bio, France社製)を一次抗体として用いた。シグナルはECL-plusウェスタンブロッティング(Amersham社製)で可視化し、X線フィルム(Konica社製)に曝露した。シグナルの定量は、膜をFluorochem(Alpha Innotech, US社製)でスキャンすることにより行った。
[Quantification of infectious prion protein]
The collected cells were lysed in 150 μl of 1 × Triton / DOC lysis buffer (0.5% tritonX-100, 0.5% deoxycholic acid, 150 mM NaCl, 25 mM Tris HCl pH 7.5, 2 mM EDTA and pepstatin, leupepsin). Protein concentration was measured by BCS assay (Pierce) and each sample was normalized to 2 mg protein. To analyze PrPsc production, the protein was digested with proteinase K at a concentration of 10 μg / mg for 30 minutes at 37 ° C. Digestion was inhibited with PMSF (2 mM) and samples were subjected to SDS / PAGE using a 15% SDS / PAGE gel. The protein was then transferred to a PVDF membrane (Immobilon-P, Amersham, USA). Anti-mouse PrP antibody (SS28) was used for PrPres. For detection of PrPC, SAF32 antibody (Spi-Bio, France) was used as the primary antibody. The signal was visualized by ECL-plus Western blotting (Amersham) and exposed to X-ray film (Konica). Signal quantification was performed by scanning the membrane with Fluorochem (Alpha Innotech, US).
これらの結果を図1のグラフに示す。図1中、「DMSO0.1%」は、化合物を添加しなかった場合、「5G-C-5LA」は、実施例1で合成した化合物を添加した場合、「GN8」は、特開2005−120002号公報に開示されている2−ピロリジン−1−イル−N−[4−[4−(2−ピロリジン−1−イル−アセチルアミノ)−ベンジル]−フェニル]−アセトアミドを添加した場合の感染型プリオンタンパク質の生成量を相対値で示している。その結果、「DMSO0.1%」を100としたとき、GN8は15.97であったのに対し、本発明の化合物である「5G-C-5LA」は13.16であった。5G-C-5LAで72時間処理された細胞は、同化合物で処理されなかった「DMSO0.1%」と比較して、感染性プリオンタンパク質の生成量が著しく低減されており、GTFK-1細胞系列においてスクレイピー型タンパク質の産生を強く抑制することが判明した。また、5G-C-5LAは、GN8に比べ、感染性プリオンタンパク質の生成を20%以上抑制していることが確認された。 These results are shown in the graph of FIG. In FIG. 1, “DMSO 0.1%” indicates that the compound was not added, “5G-C-5LA” indicates that the compound synthesized in Example 1 was added, and “GN8” Infection with the addition of 2-pyrrolidin-1-yl-N- [4- [4- (2-pyrrolidin-1-yl-acetylamino) -benzyl] -phenyl] -acetamide disclosed in 120002 The production amount of the type prion protein is shown as a relative value. As a result, when “DMSO 0.1%” was taken as 100, GN8 was 15.97, whereas “5G-C-5LA” as the compound of the present invention was 13.16. Cells treated with 5G-C-5LA for 72 hours had significantly reduced infectious prion protein production compared to DMSO0.1%, which was not treated with the same compound, and GT FK-1 It was found that the production of scrapie protein was strongly suppressed in the cell lineage. In addition, it was confirmed that 5G-C-5LA suppressed the production of infectious prion protein by 20% or more compared to GN8.
本発明の化合物によれば、プリオン病の予防及び治療に有用な医薬の開発を効率よく行うことができる。 According to the compound of the present invention, it is possible to efficiently develop a drug useful for the prevention and treatment of prion diseases.
Claims (8)
(式中、R1〜R4は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、又は置換若しくは無置換アミノ基を示す。R5〜R12は、同一又は異なって、水素原子、ハロゲン原子、置換基を有していてもよい炭化水素基、置換基を有していてもよい複素環式基、カルボキシル基、置換オキシカルボニル基、置換若しくは無置換カルバモイル基、シアノ基、アシル基、ニトロ基、硫黄酸基、硫黄酸エステル基、ヒドロキシル基、置換オキシ基、メルカプト基、置換チオ基、又は置換若しくは無置換アミノ基を示す。Xは単結合又は連結基を示す。環Z1及び環Z2は、それぞれ置換基を有していてもよい窒素原子含有環を示す。但し、R1〜R4の少なくとも一つは水素原子以外の基を示す)
で表される化合物。 Following formula (1)
(Wherein R 1 to R 4 are the same or different and are a hydrogen atom, a halogen atom, an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, carboxyl Group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercapto group, substituted thio group, or substituted or unsubstituted R 5 to R 12 are the same or different and each represents a hydrogen atom, a halogen atom, a hydrocarbon group which may have a substituent, or a heterocyclic group which may have a substituent. , Carboxyl group, substituted oxycarbonyl group, substituted or unsubstituted carbamoyl group, cyano group, acyl group, nitro group, sulfur acid group, sulfur acid ester group, hydroxyl group, substituted oxy group, mercap Group, a substituted thio group, or .X showing a substituted or unsubstituted amino group is a single bond or a linking group. Ring Z 1 and the ring Z 2 is a good nitrogen atom-containing ring which may have a substituent Provided that at least one of R 1 to R 4 represents a group other than a hydrogen atom)
A compound represented by
で表される化合物。 Following formula (2)
A compound represented by
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2007178247A JP2009013126A (en) | 2007-07-06 | 2007-07-06 | Prion protein structure transformation inhibitor |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2007178247A JP2009013126A (en) | 2007-07-06 | 2007-07-06 | Prion protein structure transformation inhibitor |
Publications (1)
Publication Number | Publication Date |
---|---|
JP2009013126A true JP2009013126A (en) | 2009-01-22 |
Family
ID=40354461
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2007178247A Pending JP2009013126A (en) | 2007-07-06 | 2007-07-06 | Prion protein structure transformation inhibitor |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP2009013126A (en) |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010131717A1 (en) * | 2009-05-14 | 2010-11-18 | 国立大学法人岐阜大学 | Prion protein structure transformation inhibitor and utilization of same |
JP2011063574A (en) * | 2009-09-21 | 2011-03-31 | Gifu Univ | Isotope labeled compound and isotope labeled compound precursor |
WO2015119111A1 (en) * | 2014-02-10 | 2015-08-13 | 国立大学法人岐阜大学 | Maleate of anti-prion compound, method for producing same, and pharmaceutical composition thereof |
JP2016166159A (en) * | 2015-03-10 | 2016-09-15 | 一夫 桑田 | Program and support method |
WO2021090911A1 (en) | 2019-11-08 | 2021-05-14 | セントラル硝子株式会社 | Method for selectively killing protein aggregate-containing cells, kit therefor, therapeutic drug for protein misfolding diseases and drug product for removing protein aggregates from blood product |
-
2007
- 2007-07-06 JP JP2007178247A patent/JP2009013126A/en active Pending
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010131717A1 (en) * | 2009-05-14 | 2010-11-18 | 国立大学法人岐阜大学 | Prion protein structure transformation inhibitor and utilization of same |
JP5665089B2 (en) * | 2009-05-14 | 2015-02-04 | 国立大学法人岐阜大学 | Prion protein structure conversion inhibitor and use thereof |
JP2011063574A (en) * | 2009-09-21 | 2011-03-31 | Gifu Univ | Isotope labeled compound and isotope labeled compound precursor |
WO2015119111A1 (en) * | 2014-02-10 | 2015-08-13 | 国立大学法人岐阜大学 | Maleate of anti-prion compound, method for producing same, and pharmaceutical composition thereof |
JPWO2015119111A1 (en) * | 2014-02-10 | 2017-03-23 | 国立大学法人岐阜大学 | Maleate of anti-prion compound, method for producing the same, and pharmaceutical composition thereof |
US9809563B2 (en) | 2014-02-10 | 2017-11-07 | Gifu University | Maleic acid salt of anti-prion compound, method for producing the same and pharmaceutical composition of the same |
JP2016166159A (en) * | 2015-03-10 | 2016-09-15 | 一夫 桑田 | Program and support method |
WO2021090911A1 (en) | 2019-11-08 | 2021-05-14 | セントラル硝子株式会社 | Method for selectively killing protein aggregate-containing cells, kit therefor, therapeutic drug for protein misfolding diseases and drug product for removing protein aggregates from blood product |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
TW201436790A (en) | Deuterium substituted fumarate derivatives | |
UA81186C2 (en) | N-heterocyclylmethyl benzamide derivatives, preparation thereof and use in therapy | |
TWI396688B (en) | Piperidine or piperazine substituted tetrahydro-naphthalene-1-carboxylic acid mtp inhibiting compounds | |
AU2015345257A1 (en) | 2-amino-3,5-difluoro-3,6-dimethyl-6-phenyl-3,4,5,6-tetrahydropyridines as BACE1 inhibitors for treating Alzheimer's disease | |
JP2009013126A (en) | Prion protein structure transformation inhibitor | |
EP4089102A1 (en) | Modulators of sortilin activity | |
JP6302929B2 (en) | Hydroxy aliphatic substituted phenylaminoalkyl ether derivatives | |
EP4079748A1 (en) | Modulators of sortilin activity | |
AU2016267872B2 (en) | Heterocyclicalkyl derivative compounds as selective histone deacetylase inhibitors and pharmaceutical compositions comprising the same | |
Tonali et al. | Structure-activity relationships of β-hairpin mimics as modulators of amyloid β-peptide aggregation | |
AU2014292064B2 (en) | Piperazine substituted bridged spiro[2.4]heptane derivatives as alx receptor agonists | |
CA3159182A1 (en) | Cell-permeable cyclic peptides and uses thereof | |
Cavalier et al. | Small molecule inhibitors of Ca2+-S100B reveal two protein conformations | |
US20030232815A1 (en) | Non-peptidic cyclophilin binding compounds and their use | |
JP5665089B2 (en) | Prion protein structure conversion inhibitor and use thereof | |
Van Manh et al. | Discovery of highly potent human glutaminyl cyclase (QC) inhibitors as anti-Alzheimer's agents by the combination of pharmacophore-based and structure-based design | |
Sitka et al. | Synthesis of N-substituted acyclic β-amino acids and their investigation as GABA uptake inhibitors | |
CN113264859A (en) | Naphthalene sulfonamide isothiocyanate bifunctional micromolecules as well as preparation method and application thereof | |
Yan et al. | Amyloidogenic immunoglobulin light chain kinetic stabilizers comprising a simple urea linker module reveal a novel binding sub-site | |
US6180796B1 (en) | Sulfonamide derivatives | |
US20150031892A1 (en) | Sphingolipid metabolite mimetics | |
US20180305404A1 (en) | Novel ligands for prevention of neurotoxicity of the alzheimer's disease related amyloid-beta peptide | |
Prashad et al. | Process Development of (2-Nitrophenylcarbamoyl)-(S)-prolyl-(S)-3-(2-naphthyl) alanyl-N-benzyl-N-methylamide (SDZ NKT343) | |
US20230399361A1 (en) | Macrocyclic peptides | |
JP5755821B2 (en) | MGLU2 / 3 agonist |