ITRM20060518A1 - ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE - Google Patents
ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE Download PDFInfo
- Publication number
- ITRM20060518A1 ITRM20060518A1 IT000518A ITRM20060518A ITRM20060518A1 IT RM20060518 A1 ITRM20060518 A1 IT RM20060518A1 IT 000518 A IT000518 A IT 000518A IT RM20060518 A ITRM20060518 A IT RM20060518A IT RM20060518 A1 ITRM20060518 A1 IT RM20060518A1
- Authority
- IT
- Italy
- Prior art keywords
- isoindole
- formula
- quinoxaline
- general formula
- compounds
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims description 14
- 230000000259 anti-tumor effect Effects 0.000 title claims description 13
- 238000004519 manufacturing process Methods 0.000 title claims description 13
- 150000001875 compounds Chemical class 0.000 claims description 63
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 39
- AELMKBWCGIBEEE-UHFFFAOYSA-N C1=CC=CC2=CNC=C21.N1=CC=NC2=CC=CC=C21 Chemical class C1=CC=CC2=CNC=C21.N1=CC=NC2=CC=CC=C21 AELMKBWCGIBEEE-UHFFFAOYSA-N 0.000 claims description 39
- 229910052739 hydrogen Inorganic materials 0.000 claims description 32
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 20
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 20
- -1 Isoindole-quinoxaline compound Chemical class 0.000 claims description 17
- 125000004432 carbon atom Chemical group C* 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 14
- 238000002360 preparation method Methods 0.000 claims description 14
- 239000001257 hydrogen Substances 0.000 claims description 13
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 10
- 235000019253 formic acid Nutrition 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 10
- 239000011541 reaction mixture Substances 0.000 claims description 10
- 238000001914 filtration Methods 0.000 claims description 9
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 9
- 239000002244 precipitate Substances 0.000 claims description 9
- 238000010992 reflux Methods 0.000 claims description 8
- 125000003545 alkoxy group Chemical group 0.000 claims description 7
- 125000000217 alkyl group Chemical group 0.000 claims description 7
- 125000004435 hydrogen atom Chemical group [H]* 0.000 claims description 7
- 230000001093 anti-cancer Effects 0.000 claims description 5
- 125000005843 halogen group Chemical group 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 125000000623 heterocyclic group Chemical group 0.000 claims description 5
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical group [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
- 125000003342 alkenyl group Chemical group 0.000 claims description 4
- 125000000304 alkynyl group Chemical group 0.000 claims description 4
- 125000003118 aryl group Chemical group 0.000 claims description 4
- 239000000460 chlorine Chemical group 0.000 claims description 4
- 229910052801 chlorine Chemical group 0.000 claims description 4
- GRVDJDISBSALJP-UHFFFAOYSA-N methyloxidanyl Chemical group [O]C GRVDJDISBSALJP-UHFFFAOYSA-N 0.000 claims description 4
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000002671 adjuvant Substances 0.000 claims description 3
- 125000004122 cyclic group Chemical group 0.000 claims description 3
- 229910052736 halogen Inorganic materials 0.000 claims description 3
- 150000002367 halogens Chemical class 0.000 claims description 3
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 claims description 3
- HQDSNEYSBFKNMX-UHFFFAOYSA-N 6H-isoindolo[5,4-f]quinoxaline Chemical class C12=CC=C3NC=CN=C3C1=CC=C1C2=CN=C1 HQDSNEYSBFKNMX-UHFFFAOYSA-N 0.000 claims description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 2
- 229910003827 NRaRb Inorganic materials 0.000 claims description 2
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 2
- 150000002431 hydrogen Chemical group 0.000 claims 4
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 32
- 239000000543 intermediate Substances 0.000 description 29
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 24
- 210000004027 cell Anatomy 0.000 description 22
- XSCHRSMBECNVNS-UHFFFAOYSA-N quinoxaline Chemical class N1=CC=NC2=CC=CC=C21 XSCHRSMBECNVNS-UHFFFAOYSA-N 0.000 description 19
- 230000000694 effects Effects 0.000 description 16
- 239000007787 solid Substances 0.000 description 16
- 229910052799 carbon Inorganic materials 0.000 description 14
- 229910052757 nitrogen Inorganic materials 0.000 description 14
- 102000029749 Microtubule Human genes 0.000 description 13
- 108091022875 Microtubule Proteins 0.000 description 13
- 230000015572 biosynthetic process Effects 0.000 description 13
- 150000003252 quinoxalines Chemical class 0.000 description 13
- 210000004688 microtubule Anatomy 0.000 description 12
- 239000002246 antineoplastic agent Substances 0.000 description 11
- 238000000338 in vitro Methods 0.000 description 11
- 238000003786 synthesis reaction Methods 0.000 description 11
- VHMICKWLTGFITH-UHFFFAOYSA-N 2H-isoindole Chemical compound C1=CC=CC2=CNC=C21 VHMICKWLTGFITH-UHFFFAOYSA-N 0.000 description 10
- 206010028980 Neoplasm Diseases 0.000 description 9
- 210000004881 tumor cell Anatomy 0.000 description 9
- 238000003556 assay Methods 0.000 description 8
- 238000003818 flash chromatography Methods 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 238000012360 testing method Methods 0.000 description 8
- 102000003915 DNA Topoisomerases Human genes 0.000 description 7
- 108090000323 DNA Topoisomerases Proteins 0.000 description 7
- 102000004243 Tubulin Human genes 0.000 description 7
- 108090000704 Tubulin Proteins 0.000 description 7
- GEYOCULIXLDCMW-UHFFFAOYSA-N 1,2-phenylenediamine Chemical compound NC1=CC=CC=C1N GEYOCULIXLDCMW-UHFFFAOYSA-N 0.000 description 6
- 206010006187 Breast cancer Diseases 0.000 description 6
- 208000026310 Breast neoplasm Diseases 0.000 description 6
- 102000007537 Type II DNA Topoisomerases Human genes 0.000 description 6
- 108010046308 Type II DNA Topoisomerases Proteins 0.000 description 6
- 238000011156 evaluation Methods 0.000 description 6
- 238000002474 experimental method Methods 0.000 description 6
- 208000032839 leukemia Diseases 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- HWNOPPADMWZJNT-UHFFFAOYSA-N 2-(2-aminophenyl)isoindole-1-carbonitrile Chemical class NC1=CC=CC=C1N1C(C#N)=C2C=CC=CC2=C1 HWNOPPADMWZJNT-UHFFFAOYSA-N 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 230000001028 anti-proliverative effect Effects 0.000 description 5
- 239000005457 ice water Substances 0.000 description 5
- 239000003112 inhibitor Substances 0.000 description 5
- 238000006116 polymerization reaction Methods 0.000 description 5
- 239000007858 starting material Substances 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 4
- IIBGKFHTRQDKCG-UHFFFAOYSA-N 2-(2-amino-4-methoxyphenyl)isoindole-1-carbonitrile Chemical compound NC1=CC(OC)=CC=C1N1C(C#N)=C2C=CC=CC2=C1 IIBGKFHTRQDKCG-UHFFFAOYSA-N 0.000 description 4
- 206010009944 Colon cancer Diseases 0.000 description 4
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 4
- 230000000118 anti-neoplastic effect Effects 0.000 description 4
- 229940041181 antineoplastic drug Drugs 0.000 description 4
- 208000029742 colonic neoplasm Diseases 0.000 description 4
- 230000008034 disappearance Effects 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 239000003480 eluent Substances 0.000 description 4
- 125000000956 methoxy group Chemical group [H]C([H])([H])O* 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- MQLACMBJVPINKE-UHFFFAOYSA-N 10-[(3-hydroxy-4-methoxyphenyl)methylidene]anthracen-9-one Chemical compound C1=C(O)C(OC)=CC=C1C=C1C2=CC=CC=C2C(=O)C2=CC=CC=C21 MQLACMBJVPINKE-UHFFFAOYSA-N 0.000 description 3
- XWXIKMKQVLMNFT-UHFFFAOYSA-N 2-(2-amino-3-methylphenyl)isoindole-1-carbonitrile Chemical compound CC1=CC=CC(N2C(=C3C=CC=CC3=C2)C#N)=C1N XWXIKMKQVLMNFT-UHFFFAOYSA-N 0.000 description 3
- 239000011543 agarose gel Substances 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 229940034982 antineoplastic agent Drugs 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- ZMMJGEGLRURXTF-UHFFFAOYSA-N ethidium bromide Chemical compound [Br-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 ZMMJGEGLRURXTF-UHFFFAOYSA-N 0.000 description 3
- 229960005542 ethidium bromide Drugs 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 201000001441 melanoma Diseases 0.000 description 3
- 230000000394 mitotic effect Effects 0.000 description 3
- 230000003389 potentiating effect Effects 0.000 description 3
- XWACOSRTCVIMQE-UHFFFAOYSA-N 1-methoxy-2h-isoindole Chemical compound C1=CC=CC2=C(OC)NC=C21 XWACOSRTCVIMQE-UHFFFAOYSA-N 0.000 description 2
- VCMZTDGLGQTLFQ-UHFFFAOYSA-N 4-methyl-2h-isoindole Chemical compound CC1=CC=CC2=CNC=C12 VCMZTDGLGQTLFQ-UHFFFAOYSA-N 0.000 description 2
- 102100033350 ATP-dependent translocase ABCB1 Human genes 0.000 description 2
- 102000014914 Carrier Proteins Human genes 0.000 description 2
- 108010078791 Carrier Proteins Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 102000001267 GSK3 Human genes 0.000 description 2
- 108010014905 Glycogen Synthase Kinase 3 Proteins 0.000 description 2
- 101000830681 Homo sapiens DNA topoisomerase 1 Proteins 0.000 description 2
- 208000008839 Kidney Neoplasms Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 108010047230 Member 1 Subfamily B ATP Binding Cassette Transporter Proteins 0.000 description 2
- 206010033128 Ovarian cancer Diseases 0.000 description 2
- 206010061535 Ovarian neoplasm Diseases 0.000 description 2
- 206010060862 Prostate cancer Diseases 0.000 description 2
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 2
- KAESVJOAVNADME-UHFFFAOYSA-N Pyrrole Chemical compound C=1C=CNC=1 KAESVJOAVNADME-UHFFFAOYSA-N 0.000 description 2
- 206010038389 Renal cancer Diseases 0.000 description 2
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 239000005557 antagonist Substances 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000000843 anti-fungal effect Effects 0.000 description 2
- 230000000845 anti-microbial effect Effects 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- DIKBFYAXUHHXCS-UHFFFAOYSA-N bromoform Chemical compound BrC(Br)Br DIKBFYAXUHHXCS-UHFFFAOYSA-N 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 102000007588 cdc25 Phosphatases Human genes 0.000 description 2
- 108010046616 cdc25 Phosphatases Proteins 0.000 description 2
- 208000025997 central nervous system neoplasm Diseases 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229960001338 colchicine Drugs 0.000 description 2
- 210000004292 cytoskeleton Anatomy 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000002489 hematologic effect Effects 0.000 description 2
- 102000056859 human TOP1 Human genes 0.000 description 2
- 238000010166 immunofluorescence Methods 0.000 description 2
- 238000010348 incorporation 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
- 230000002401 inhibitory effect Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 201000010982 kidney cancer Diseases 0.000 description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 2
- 150000004987 o-phenylenediamines Chemical class 0.000 description 2
- 230000007170 pathology Effects 0.000 description 2
- 239000000546 pharmaceutical excipient Substances 0.000 description 2
- NNFCIKHAZHQZJG-UHFFFAOYSA-N potassium cyanide Chemical compound [K+].N#[C-] NNFCIKHAZHQZJG-UHFFFAOYSA-N 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- FFRYUAVNPBUEIC-UHFFFAOYSA-N quinoxalin-2-ol Chemical class C1=CC=CC2=NC(O)=CN=C21 FFRYUAVNPBUEIC-UHFFFAOYSA-N 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229940079827 sodium hydrogen sulfite Drugs 0.000 description 2
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 2
- 125000001424 substituent group Chemical group 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 2
- MDMDITFPFDTNAD-VIFPVBQESA-N (4S)-4-amino-5-benzoyloxy-5-oxopentanoic acid Chemical class OC(=O)CC[C@H](N)C(=O)OC(=O)C1=CC=CC=C1 MDMDITFPFDTNAD-VIFPVBQESA-N 0.000 description 1
- FMCUPJKTGNBGEC-UHFFFAOYSA-N 1,2,4-triazol-4-amine Chemical compound NN1C=NN=C1 FMCUPJKTGNBGEC-UHFFFAOYSA-N 0.000 description 1
- KDWOWEWZOWGWMH-UHFFFAOYSA-N 1,2-dimethylisoindole Chemical compound C1=CC=CC2=C(C)N(C)C=C21 KDWOWEWZOWGWMH-UHFFFAOYSA-N 0.000 description 1
- QLXHAGHQLRUWLX-UHFFFAOYSA-N 1-n,2-n-dichlorobenzene-1,2-diamine Chemical compound ClNC1=CC=CC=C1NCl QLXHAGHQLRUWLX-UHFFFAOYSA-N 0.000 description 1
- UMAPFAAAQBMYNJ-UHFFFAOYSA-N 1-n,2-n-dimethylbenzene-1,2-diamine Chemical compound CNC1=CC=CC=C1NC UMAPFAAAQBMYNJ-UHFFFAOYSA-N 0.000 description 1
- DWGMRQGGIJRVMF-UHFFFAOYSA-N 1h-benzimidazolo[4,5-h]isoquinoline Chemical compound C1=CC2=CC=NC=C2C2=C1C(N=CN1)=C1C=C2 DWGMRQGGIJRVMF-UHFFFAOYSA-N 0.000 description 1
- PBUOWEVOJVKQGA-UHFFFAOYSA-N 1h-pyrido[2,3-g]quinoxalin-2-one Chemical class C1=CC=C2C=C(NC(=O)C=N3)C3=CC2=N1 PBUOWEVOJVKQGA-UHFFFAOYSA-N 0.000 description 1
- UHZLPLRECKEPAX-UHFFFAOYSA-N 1h-pyrido[2,3-h]quinoxalin-2-one Chemical class C1=CC=C2C3=NC(O)=CN=C3C=CC2=N1 UHZLPLRECKEPAX-UHFFFAOYSA-N 0.000 description 1
- BDZHCMDWAXSQMS-UHFFFAOYSA-N 1h-pyrrolo[2,1-d][1,2,3,5]tetrazin-4-one Chemical class O=C1NN=NC2=CC=CN12 BDZHCMDWAXSQMS-UHFFFAOYSA-N 0.000 description 1
- JMWDPGBZVXOLNK-UHFFFAOYSA-N 2-(2-amino-4,5-dimethylphenyl)isoindole-1-carbonitrile Chemical compound C1=C(C)C(C)=CC(N)=C1N1C(C#N)=C2C=CC=CC2=C1 JMWDPGBZVXOLNK-UHFFFAOYSA-N 0.000 description 1
- IDBPRPHZTRUDAE-UHFFFAOYSA-N 2-(6-amino-5,6-dichlorocyclohexa-2,4-dien-1-yl)isoindole-1-carbonitrile Chemical compound C(#N)C=1N(C=C2C=CC=CC12)C1C(C(=CC=C1)Cl)(Cl)N IDBPRPHZTRUDAE-UHFFFAOYSA-N 0.000 description 1
- FIZQBOULTBAUBU-UHFFFAOYSA-N 2-(6-amino-5,6-dimethylcyclohexa-2,4-dien-1-yl)isoindole-1-carbonitrile Chemical compound C(#N)C=1N(C=C2C=CC=CC12)C1C(C(=CC=C1)C)(C)N FIZQBOULTBAUBU-UHFFFAOYSA-N 0.000 description 1
- ZPRQXVPYQGBZON-UHFFFAOYSA-N 2-bromo-1h-indole Chemical compound C1=CC=C2NC(Br)=CC2=C1 ZPRQXVPYQGBZON-UHFFFAOYSA-N 0.000 description 1
- CVOFKRWYWCSDMA-UHFFFAOYSA-N 2-chloro-n-(2,6-diethylphenyl)-n-(methoxymethyl)acetamide;2,6-dinitro-n,n-dipropyl-4-(trifluoromethyl)aniline Chemical compound CCC1=CC=CC(CC)=C1N(COC)C(=O)CCl.CCCN(CCC)C1=C([N+]([O-])=O)C=C(C(F)(F)F)C=C1[N+]([O-])=O CVOFKRWYWCSDMA-UHFFFAOYSA-N 0.000 description 1
- KJAAWBYLNVBKPI-UHFFFAOYSA-N 2-n-methoxybenzene-1,2-diamine Chemical compound CONC1=CC=CC=C1N KJAAWBYLNVBKPI-UHFFFAOYSA-N 0.000 description 1
- RPKCLSMBVQLWIN-UHFFFAOYSA-N 2-n-methylbenzene-1,2-diamine Chemical compound CNC1=CC=CC=C1N RPKCLSMBVQLWIN-UHFFFAOYSA-N 0.000 description 1
- FCMCRFMYSONEAN-UHFFFAOYSA-N 2-phenoxy-2-quinoxalin-2-yloxypropanoic acid Chemical class C=1N=C2C=CC=CC2=NC=1OC(C(O)=O)(C)OC1=CC=CC=C1 FCMCRFMYSONEAN-UHFFFAOYSA-N 0.000 description 1
- VYYVURMFAYDYFT-UHFFFAOYSA-N 3-(phenoxymethyl)-1h-quinoxalin-2-one Chemical class O=C1NC2=CC=CC=C2N=C1COC1=CC=CC=C1 VYYVURMFAYDYFT-UHFFFAOYSA-N 0.000 description 1
- YELMWJNXDALKFE-UHFFFAOYSA-N 3h-imidazo[4,5-f]quinoxaline Chemical class N1=CC=NC2=C(NC=N3)C3=CC=C21 YELMWJNXDALKFE-UHFFFAOYSA-N 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- 208000023275 Autoimmune disease Diseases 0.000 description 1
- SYUVJSJBRJSPLV-UHFFFAOYSA-N Benzimidazo[2,1-a]isoquinoline Chemical compound C1=CC=C2C=CN3C4=CC=CC=C4N=C3C2=C1 SYUVJSJBRJSPLV-UHFFFAOYSA-N 0.000 description 1
- 101150012716 CDK1 gene Proteins 0.000 description 1
- KLWPJMFMVPTNCC-UHFFFAOYSA-N Camptothecin Natural products CCC1(O)C(=O)OCC2=C1C=C3C4Nc5ccccc5C=C4CN3C2=O KLWPJMFMVPTNCC-UHFFFAOYSA-N 0.000 description 1
- 239000012625 DNA intercalator Substances 0.000 description 1
- 230000007018 DNA scission Effects 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 101100059559 Emericella nidulans (strain FGSC A4 / ATCC 38163 / CBS 112.46 / NRRL 194 / M139) nimX gene Proteins 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- QTANTQQOYSUMLC-UHFFFAOYSA-O Ethidium cation Chemical compound C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CC)=C1C1=CC=CC=C1 QTANTQQOYSUMLC-UHFFFAOYSA-O 0.000 description 1
- LELOWRISYMNNSU-UHFFFAOYSA-N Hydrocyanic acid Natural products N#C LELOWRISYMNNSU-UHFFFAOYSA-N 0.000 description 1
- 102100021339 Multidrug resistance-associated protein 1 Human genes 0.000 description 1
- 206010048723 Multiple-drug resistance Diseases 0.000 description 1
- KSLNFJQTUZNRHC-UHFFFAOYSA-N N1=CC=C2N=C3C4=CC=CC=C4C=CC3=NC2=C1 Chemical class N1=CC=C2N=C3C4=CC=CC=C4C=CC3=NC2=C1 KSLNFJQTUZNRHC-UHFFFAOYSA-N 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 108091000080 Phosphotransferase Proteins 0.000 description 1
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical group C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- BPEGJWRSRHCHSN-UHFFFAOYSA-N Temozolomide Chemical compound O=C1N(C)N=NC2=C(C(N)=O)N=CN21 BPEGJWRSRHCHSN-UHFFFAOYSA-N 0.000 description 1
- 101710183280 Topoisomerase Proteins 0.000 description 1
- JXLYSJRDGCGARV-WWYNWVTFSA-N Vinblastine Natural products O=C(O[C@H]1[C@](O)(C(=O)OC)[C@@H]2N(C)c3c(cc(c(OC)c3)[C@]3(C(=O)OC)c4[nH]c5c(c4CCN4C[C@](O)(CC)C[C@H](C3)C4)cccc5)[C@@]32[C@H]2[C@@]1(CC)C=CCN2CC3)C JXLYSJRDGCGARV-WWYNWVTFSA-N 0.000 description 1
- 101100273808 Xenopus laevis cdk1-b gene Proteins 0.000 description 1
- 150000001251 acridines Chemical class 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 229940100198 alkylating agent Drugs 0.000 description 1
- 239000002168 alkylating agent Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 229940045799 anthracyclines and related substance Drugs 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 238000002841 anti-cancer assay Methods 0.000 description 1
- 230000000340 anti-metabolite Effects 0.000 description 1
- 230000003377 anti-microbal effect Effects 0.000 description 1
- 238000012460 anticancer assay Methods 0.000 description 1
- 239000003429 antifungal agent Substances 0.000 description 1
- 229940100197 antimetabolite Drugs 0.000 description 1
- 239000002256 antimetabolite Substances 0.000 description 1
- 239000003080 antimitotic agent Substances 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- DCBDOYDVQJVXOH-UHFFFAOYSA-N azane;1h-indole Chemical compound N.C1=CC=C2NC=CC2=C1 DCBDOYDVQJVXOH-UHFFFAOYSA-N 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 125000005605 benzo group Chemical group 0.000 description 1
- 125000002619 bicyclic group Chemical group 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 229950005228 bromoform Drugs 0.000 description 1
- VSJKWCGYPAHWDS-FQEVSTJZSA-N camptothecin Chemical compound C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)[C@]5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-FQEVSTJZSA-N 0.000 description 1
- 229940127093 camptothecin Drugs 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 201000007455 central nervous system cancer Diseases 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 125000000259 cinnolinyl group Chemical class N1=NC(=CC2=CC=CC=C12)* 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 125000004093 cyano group Chemical group *C#N 0.000 description 1
- 229940127089 cytotoxic agent Drugs 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- VSJKWCGYPAHWDS-UHFFFAOYSA-N dl-camptothecin Natural products C1=CC=C2C=C(CN3C4=CC5=C(C3=O)COC(=O)C5(O)CC)C4=NC2=C1 VSJKWCGYPAHWDS-UHFFFAOYSA-N 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 238000007876 drug discovery Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000009422 growth inhibiting effect Effects 0.000 description 1
- 230000009036 growth inhibition Effects 0.000 description 1
- 201000005787 hematologic cancer Diseases 0.000 description 1
- 208000024200 hematopoietic and lymphoid system neoplasm Diseases 0.000 description 1
- 150000002391 heterocyclic compounds Chemical class 0.000 description 1
- 238000007074 heterocyclization reaction Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 150000002466 imines Chemical group 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000005918 in vitro anti-tumor Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- XVLVNGLXPVWGNV-UHFFFAOYSA-N indolo[1,2-c][1,2,3]benzotriazine Chemical compound N12N=NC3=CC=C[CH]C3=C2C=C2[C]1C=CC=C2 XVLVNGLXPVWGNV-UHFFFAOYSA-N 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000002687 intercalation Effects 0.000 description 1
- 238000009830 intercalation Methods 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 210000005265 lung cell Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000025090 microtubule depolymerization Effects 0.000 description 1
- 230000029115 microtubule polymerization Effects 0.000 description 1
- QXYYYPFGTSJXNS-UHFFFAOYSA-N mitozolomide Chemical compound N1=NN(CCCl)C(=O)N2C1=C(C(=O)N)N=C2 QXYYYPFGTSJXNS-UHFFFAOYSA-N 0.000 description 1
- 229950005967 mitozolomide Drugs 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 108010066052 multidrug resistance-associated protein 1 Proteins 0.000 description 1
- 210000005170 neoplastic cell Anatomy 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- 125000004433 nitrogen atom Chemical group N* 0.000 description 1
- 229910017464 nitrogen compound Inorganic materials 0.000 description 1
- 150000002830 nitrogen compounds Chemical class 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 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
- 230000002018 overexpression Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 102000020233 phosphotransferase Human genes 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 125000003367 polycyclic group Chemical group 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000002685 pulmonary effect Effects 0.000 description 1
- RAPMCFNYFAPCGO-UHFFFAOYSA-N pyrido[4,3-b]quinoxaline Chemical class C1=NC=CC2=NC3=CC=CC=C3N=C21 RAPMCFNYFAPCGO-UHFFFAOYSA-N 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- NAPZETGQDAEAEM-UHFFFAOYSA-N quinoxaline;2h-triazole Chemical class C1=CNN=N1.N1=CC=NC2=CC=CC=C21 NAPZETGQDAEAEM-UHFFFAOYSA-N 0.000 description 1
- 125000001567 quinoxalinyl group Chemical group N1=C(C=NC2=CC=CC=C12)* 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- RWWYLEGWBNMMLJ-YSOARWBDSA-N remdesivir Chemical compound NC1=NC=NN2C1=CC=C2[C@]1([C@@H]([C@@H]([C@H](O1)CO[P@](=O)(OC1=CC=CC=C1)N[C@H](C(=O)OCC(CC)CC)C)O)O)C#N RWWYLEGWBNMMLJ-YSOARWBDSA-N 0.000 description 1
- 230000004043 responsiveness Effects 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 229960004964 temozolomide Drugs 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 229960003048 vinblastine Drugs 0.000 description 1
- JXLYSJRDGCGARV-XQKSVPLYSA-N vincaleukoblastine Chemical compound C([C@@H](C[C@]1(C(=O)OC)C=2C(=CC3=C([C@]45[C@H]([C@@]([C@H](OC(C)=O)[C@]6(CC)C=CCN([C@H]56)CC4)(O)C(=O)OC)N3C)C=2)OC)C[C@@](C2)(O)CC)N2CCC2=C1NC1=CC=CC=C21 JXLYSJRDGCGARV-XQKSVPLYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
- C07D487/04—Ortho-condensed systems
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
Description
DESCRIZIONE DESCRIPTION
“Derivati isoindolo-chinossalinici ad attività antitumorale, procedimento per la loro produzione e loro uso” "Isoindole-quinoxaline derivatives with antitumor activity, procedure for their production and their use"
La presente invenzione riguarda nuovi derivati isoindolochinossalinici ad attività antitumorale, un procedimento per la loro produzione e il loro uso in campo medico. Più in particolare, l'invenzione concerne la sintesi e lo studio di derivati isoindolo[2,1-a]chinossalinici che hanno rivelato una potente attività antitumorale in vitro, e vengono quindi proposti per la produzione di farmaci da impiegare nella terapia dei tumori, sia da soli che in combinazione con altri chemioterapici. The present invention relates to new isoindoloquinoxaline derivatives with antitumor activity, a process for their production and their use in the medical field. More specifically, the invention concerns the synthesis and study of isoindole [2,1-a] quinoxaline derivatives which have revealed a powerful antitumor activity in vitro, and are therefore proposed for the production of drugs to be used in the therapy of tumors, either alone or in combination with other chemotherapy drugs.
Come è noto, il DNA rappresenta uno dei più importanti bersagli per numerosi agenti chemioterapeutici. In modo specifico, tra le molecole antitumorali note sono ampiamente diffusi, ad esempio, gli agenti alchilanti, che possono modificare non selettivamente il DNA e l'RNA delle cellule, e gli antimetaboliti, che mimando le basi del DNA e altre molecole naturali interferiscono con la sintesi degli acidi nucleici, delle proteine e con altri processi metabolici vitali, agendo come antagonisti e inibitori. Le molecole policicliche azotate a struttura planare possono agire come farmacoforì per determinate classi di farmaci antineoplastici, perché possono intercalarsi tra le coppie di basi della doppia elica del DNA. Molecole antitumorali ben note come le acridine, le amsacrine, le antracicline, caratterizzate da sistemi policiclici planari, sono in grado di interferire con gli enzimi che elaborano il DNA (topoisomerasi I e II) mediante la formazione di complessi ternari tra il farmaco, il DNA e l’enzima. As is known, DNA represents one of the most important targets for numerous chemotherapeutic agents. Specifically, among the known anticancer molecules, for example, alkylating agents, which can non-selectively modify the DNA and RNA of cells, and antimetabolites, which by mimicking the bases of DNA and other natural molecules interfere with the synthesis of nucleic acids, proteins and other vital metabolic processes, acting as antagonists and inhibitors. The polycyclic nitrogenous molecules with a planar structure can act as pharmacophores for certain classes of antineoplastic drugs, because they can intercalate between the base pairs of the DNA double helix. Well-known anticancer molecules such as acridines, amsacrines, anthracyclines, characterized by planar polycyclic systems, are able to interfere with the enzymes that process DNA (topoisomerase I and II) by forming ternary complexes between the drug, the DNA and the enzyme.
Le chinossaline o 1 ,4-benzodiazine, composti biciclici aromatici contenenti due atomi di azoto, e i chinossalinoni ad esse strutturalmente correlati, rappresentano un’importante classe di composti eterociclici che si trovano in un gran numero di agenti biologicamente e farmacologicamente attivi, e molti di essi hanno dimostrato, a vari livelli sperimentali, attività antineoplastica. Ad esempio, la domanda di brevetto internazionale pubbl. No. WO 94/13647 (cessionaria The Du Pont Merck Pharmaceutical Co.) concerne una classe di esteri di acidi chinossalinossi-fenossipropanoici che risultano attivi come agenti antineoplastici in base a sperimentazioni in vivo in modelli animali. Ancora nel campo delle strutture aromatiche bicicliche, Lawrence et al. (Lawrence, D.S.; Copper, J.E.; Smith, C.D. Structure-activity Studies of Substituted Quinoxalinones as Multiple-Drug-Resistance Antagonists. J. Med. Chem. Quinoxalines or 1,4-benzodiazines, bicyclic aromatic compounds containing two nitrogen atoms, and the structurally related quinoxalinones, represent an important class of heterocyclic compounds found in a large number of biologically and pharmacologically active agents, and many of they have demonstrated antineoplastic activity at various experimental levels. For example, the international patent application publ. No. WO 94/13647 (assignee The Du Pont Merck Pharmaceutical Co.) relates to a class of esters of quinoxalinoxy-phenoxypropanoic acids which are active as antineoplastic agents based on in vivo experiments in animal models. Still in the field of bicyclic aromatic structures, Lawrence et al. (Lawrence, D.S .; Copper, J.E .; Smith, C.D. Structure-activity Studies of Substituted Quinoxalinones as Multiple-Drug-Resistance Antagonists. J. Med. Chem.
2001, 44, 594-601) presentano una classe di fenossimetil-chinossalinoni sostituiti che hanno mostrato un eccellente antagonismo alla P-glicoproteina (una proteina di trasporto la cui sovraespressione può mediare la resistenza ai farmaci) e alla MRP1 (multidrug-related resistance protein 1 , una ulteriore proteina di trasporto che ha diverse caratteristiche in comune con la P-glicoproteina) in linee cellulari resistenti ai farmaci. 2001, 44, 594-601) present a class of substituted phenoxymethyl-quinoxalinones that showed excellent antagonism to P-glycoprotein (a transport protein whose overexpression can mediate drug resistance) and to MRP1 (multidrug-related resistance protein 1, an additional transport protein that has several features in common with P-glycoprotein) in drug-resistant cell lines.
Passando alle strutture tricicliche, ad esempio, alcuni piridochinossalinoni funzionalizzati si sono dimostrati attivi nella valutazione in vitro dell'attività antimicrobica nei confronti di un ampio spettro di microrganismi, e uno di essi ha mostrato una promettente attività antitumorale in vitro (Carta, A. et al. Novel functionalized pyrido[2,3-g]quinoxalinones as antibacterial, antifungal and anticancer agents, Il Farmaco, 2001, 56, 933-938). Analogamente, alcune pirrolo[1 ,2-a]chinossaline sono risultate dotate di una potente attività antiproliferativa su linee cellulari derivate da tumori ematologici e solidi (Alleca S. et al Quinoxaline chemistry. Part 16. 4-Substituted anilino and 4-substituted phenoxymethyl pyrrolo[1,2-a]quinoxalines and N-[4-(pyrrolo[1 ,2-a]quinoxalin-4-yl)amino and hydroxymethyl]benzoyl glutamates. Synthesis and evaluation of in vitro biological activity. Il Farmaco, 2003, 58, 639-650). Turning to tricyclic structures, for example, some functionalized pyridoquinoxalones have been shown to be active in the in vitro evaluation of antimicrobial activity against a broad spectrum of microorganisms, and one of them has shown a promising antitumor activity in vitro (Carta, A. et al. Novel functionalized pyrido [2,3-g] quinoxalinones as antibacterial, antifungal and anticancer agents, Il Farmaco, 2001, 56, 933-938). Similarly, some pyrrole [1, 2-a] quinoxalines were found to have potent antiproliferative activity on cell lines derived from hematological and solid tumors (Alleca S. et al Quinoxaline chemistry. Part 16. 4-Substituted anilino and 4-substituted phenoxymethyl pyrrolo [1,2-a] quinoxalines and N- [4- (pyrrolo [1, 2-a] quinoxalin-4-yl) amino and hydroxymethyl] benzoyl glutamates. Synthesis and evaluation of in vitro biological activity. Il Farmaco, 2003 , 58, 639-650).
Le imidazo-chinossaline sono state studiate da Chen e coll., che hanno riportato per una classe di anilino-imidazo[1,5-a]chinossaline una eccellente attività enzimatica nell'inibizione dell'enzima Lek (IC50<5 nM) e della proliferazione di cellule T, attività che è indicata per il trattamento di malattie autoimmuni e infiammatorie mediate da cellule T (Chen P. et. al, Synthesis and SAR of Novel Imidazoquinoxaline-Based Lek Inhibitors: Improvements of Celi Potency, Bioorg. Med. Chem. Leti., 2002, 12, 3153-3156). Come ulteriore esempio di composti azotati triciclici che sono stati proposti per la terapia antiproliferativa, la domanda di brevetto internazionale pubbl. No. WO 2004/085439 (cessionaria Pfizer Products Ine.) concerne una classe di triazolo-chinossaline sostituite (in modo specifico, 4-ammino[1,2,4]triazolo[4,3-a]chinossaline, che sono risultate attive come inibitori delle glicogeno-sintasi chinasi-3 (GSK-3) e quindi, come tali, potenzialmente utili per il trattamento di varie patologie tra cui le patologie tumorali. The imidazo-quinoxalines have been studied by Chen and coll., Who reported for an anilino-imidazo [1,5-a] quinoxalines class an excellent enzyme activity in the inhibition of the Lek enzyme (IC50 <5 nM) and of the proliferation of T cells, activity which is indicated for the treatment of autoimmune and inflammatory diseases mediated by T cells (Chen P. et. al, Synthesis and SAR of Novel Imidazoquinoxaline-Based Lek Inhibitors: Improvements of Celi Potency, Bioorg. Med. Chem . Leti., 2002, 12, 3153-3156). As a further example of tricyclic nitrogen compounds that have been proposed for antiproliferative therapy, the published international patent application No. WO 2004/085439 (assignee Pfizer Products Ine.) Relates to a class of substituted triazole-quinoxalines (specifically, 4-amino [1,2,4] triazole [4,3-a] quinoxalines, which have been found to be active as inhibitors of glycogen synthase kinase-3 (GSK-3) and therefore, as such, potentially useful for the treatment of various pathologies including tumor pathologies.
Anche derivati chinossalinici tetraciclici sono stati sintetizzati e studiati neil'ambÌto delle ricerche su composti ad attività antineoplastica. In particolare il sistema chinossalinico condensato con gruppi piridinici ha portato a benzo[/]pirido[4,3-ò]chinossaline e a benzo[f]pirido[3,4-b]chinossaline, che si sono dimostrate attive come inibitori delle topoisomerasi I e II, ed hano significative proprietà antitumorali (Nguyen, C.H. et al., Synthesis and biologica! evaluation of amino-substituted benzo[fpyrido[4,-b] and pyrido[3,4-b]quinoxalines: a new class of antineoplastic agents. Anti-Cancer Drug Design, 1995, 10, 277-297). Infine , le bromoindolo-chinossaline sostituite nell’azoto indolico (2-bromoindolo[3,2-b]chinossaline 5-sostituite) hanno mostrato un ampio spettro di attività antitumorale, con valori medi di Gl50, TGl, e LC50di 14,2, 31,6 e 66,2 μΜ rispettivamente. Inoltre, tali composti sono risultati attivi sia nell’inibizione della cdc2 chinasi che nell'inibizione della cdc25 fosfatasi, con IC50di 70 e 25 μM rispettivamente (Abadi A.H. 5-Substituted 2-Bromoindolo[3,2-b]quinoxalines. A Class of Potential Antitumor Agents with cdc25 Phosphatase Inhibitory Properties. Arch. Pharm. Pharm. Med. Chem., 1998, 331, 352-358). Tetracyclic quinoxaline derivatives have also been synthesized and studied in the context of research on compounds with antineoplastic activity. In particular, the quinoxaline system condensed with pyridine groups led to benzo [/] pyrido [4,3-ò] quinoxalines and to benzo [f] pyrido [3,4-b] quinoxalines, which proved to be active as inhibitors of topoisomerase I and II, and have significant anticancer properties (Nguyen, C.H. et al., Synthesis and biological! evaluation of amino-substituted benzo [fpyrido [4, -b] and pyrido [3,4-b] quinoxalines: a new class of antineoplastic agents. Anti-Cancer Drug Design, 1995, 10, 277-297). Finally, the bromoindole-quinoxalines substituted in indole nitrogen (2-bromoindole [3,2-b] 5-substituted quinoxalines) showed a broad spectrum of antitumor activity, with mean values of Gl50, TG1, and LC50 of 14.2, 31.6 and 66.2 μΜ respectively. Furthermore, these compounds were active both in the inhibition of cdc2 kinase and in the inhibition of cdc25 phosphatase, with IC50 of 70 and 25 μM respectively (Abadi A.H. 5-Substituted 2-Bromoindole [3,2-b] quinoxalines. A Class of Potential Antitumor Agents with cdc25 Phosphatase Inhibitory Properties. Arch. Pharm. Pharm. Med. Chem., 1998, 331, 352-358).
Nell’ambito della ricerca su strutture eterocicliche a più anelli condensati contenenti porzioni pirroliche o indoliche che fossero dotate di attività antineoplastica, condotta dal gruppo di ricerca degli attuali inventori, sono state studiate, ad esempio, la sintesi e le proprietà antimicrobiche e antiproliferative di indolo-cinnoline e di indolo-benzotrioazine, strutture tetracicliche planari, e di pirrolo-tetrazinoni, strutture bicicliche che possiedono lo stesso scheletro dei due agenti antitumorali noti temozolomide e mitozolomide (si vedano, ad esempio, Barraja, P. et al., lndolo[3,2-b]cinnolines with antiproliferative, antifungal and antibacterial activity. Bioorg. Med. Chem., 1999, 7, 1591-1596; Cirrincione, G. et al., Derivatives of thè New Ring System lndolo[1,2-c]benzo [1,2,3]triazine, with Potent Antitumor and Antimicrobal Activity. J.Med Chem., 1999, 42, 2561-2568; Diana, P. et al., Pyrrolo[2,1-d][1 ,2,3,5] tetrazine-4(3H)-ones. New class of azolo-tetrazinones with potent antitumor activity. Bioorg. Med. Chem., 2003, 11, 2371-2380). Nella linea di tale ricerca si è quindi preso in considerazione il sistema isoindolochinossalinico, e in particolare si è studiata la sintesi di nuovi derivati isoindolo-chinossalinici, con lo scopo di valutare la loro potenziale attività antiproliferiativa e i loro bersagli a livello cellulare e molecolare. Scopo della presente invenzione è pertanto quello di fornire nuovi agenti antineoplastici che siano dotati di una notevole attività sul più ampio campo possibile di tipi tumorali e la cui sintesi sia semplice da realizzare e al tempo stesso economica in relazione alle materie prime necessarie. As part of the research on heterocyclic structures with multiple condensed rings containing pyrrolic or indole portions that were endowed with antineoplastic activity, conducted by the research group of the current inventors, the synthesis and antimicrobial and antiproliferative properties of indole were studied, for example. -cinnolines and indole-benzotrioazines, planar tetracyclic structures, and pyrrole-tetrazinones, bicyclic structures that possess the same skeleton as the two known anticancer agents temozolomide and mitozolomide (see, for example, Barraja, P. et al., lndolo [ 3,2-b] cinnolines with antiproliferative, antifungal and antibacterial activity. Bioorg. Med. Chem., 1999, 7, 1591-1596; Cirrincione, G. et al., Derivatives of the New Ring System Indolo [1,2- c] benzo [1,2,3] triazine, with Potent Antitumor and Antimicrobal Activity. J. Med Chem., 1999, 42, 2561-2568; Diana, P. et al., Pyrrolo [2,1-d] [ 1, 2,3,5] tetrazine-4 (3H) -ones. New class of azole-tetrazinones with potent antitumor activity. Bioorg. Med. Chem., 2003, 11, 2371-2380). The isoindole quinoxaline system was therefore considered in the line of this research, and in particular the synthesis of new isoindole-quinoxaline derivatives was studied, with the aim of evaluating their potential antiproliferative activity and their targets at the cellular and molecular level. The object of the present invention is therefore to provide new antineoplastic agents which are endowed with a remarkable activity on the widest possible range of tumor types and whose synthesis is simple to carry out and at the same time economical in relation to the necessary raw materials.
Secondo l'invenzione è stato trovato che una classe di nuovi derivati isoindolo[2,1-a]chinossalinici, che possono essere sintetizzati con ottime rese e con semplici reazioni a partire da specifici intermedi noti in letteratura, i 2-(2’-amminoaril)-1-ciano-isoindoli (Diana, P. et al., 19<th>I.C.H.C. Aug 2003, p. 284) possiedono una notevole attività antitumorale, che è stata dimostrata in vitro mediante uno screening primario su un panel di tre linee cellulari tumorali appositamente prescelte (carcinoma polmonare a cellule non piccole, carcinoma mammario e tumore del SNC) ed è stata poi confermata attraverso uno screening in vitro completo contro circa 60 diverse linee cellulari tumorali, appartenenti a diversi tipi di tumori, sia ematologici che solidi. According to the invention it has been found that a class of new isoindole [2,1-a] quinoxaline derivatives, which can be synthesized with excellent yields and with simple reactions starting from specific intermediates known in literature, the 2- (2'- aminoaryl) -1-cyano-isoindoles (Diana, P. et al., 19 <th> I.C.H.C. Aug 2003, p. 284) possess remarkable antitumor activity, which was demonstrated in vitro by primary screening on a panel of three specially selected tumor cell lines (non-small cell lung cancer, breast cancer and CNS cancer) and was then confirmed through a comprehensive in vitro screening against about 60 different cancer cell lines, belonging to different types of tumors, both haematological and solid .
Formano pertanto oggetto specifico della presente invenzione composti isoindolo-chinossalinici di formula generale (1): Therefore, isoindole-quinoxaline compounds of general formula (1) form the specific object of the present invention:
in cui: in which:
R rappresenta H o OH e R represents H or OH e
R5rappresenta H o CN, R5 represents H or CN,
R1, R2, R3e R4sono scelti, ognuno indipendentemente dall'altro, tra H, R1, R2, R3 and R4 are chosen, each independently of the other, among H,
un gruppo alchilico, alcossilico, alchenilico o alchinilico con fino a 12 atomi di carbonio, an alkyl, alkoxy, alkenyl or alkynyl group with up to 12 carbon atoms,
un gruppo aromatico o aralchilico, isociclico o eterociclico, con fino a 12 atomi di carbonio, an aromatic or aralkyl group, isocyclic or heterocyclic, with up to 12 carbon atoms,
un atomo di alogeno, a halogen atom,
detti gruppi essendo eventualmente sostituiti con uno o più gruppi scelti tra ORa, CN, COORa, NO2, alogeni, NRaRb, in cui said groups being optionally replaced with one or more groups selected from ORa, CN, COORa, NO2, halogens, NRaRb, in which
Rae Rbsono indipendentemente scelti dal gruppo consistente in H, alchile, alchenile o alchinile lineari, ramificati o ciclici con fino a 12 atomi di carbonio, arile, eterociclo con fino a 12 atomi di carbonio. Ra and Rb are independently selected from the group consisting of linear, branched or cyclic H, alkyl, alkenyl or alkynyl with up to 12 carbon atoms, aryl, heterocycle with up to 12 carbon atoms.
Nel caso specifico in cui, nella formula (1) che precede, R rappresenta OH e R5rappresenta H, i composti secondo l'invenzione possono essere rappresentati da una o l’altra delle due strutture limite di formula (2') e (2"): In the specific case in which, in the above formula (1), R represents OH and R5 represents H, the compounds according to the invention can be represented by one or the other of the two limiting structures of formula (2 ') and (2 " ):
In questo caso, secondo alcune forme di realizzazione preferite dell'invenzione, R1,R2,R3e R4possono essere scelti, ognuno indipendentemente dall’altro, tra H, un gruppo alchilico con fino a 6 atomi di carbonio, un gruppo alcossilico con fino a 6 atomi di carbonio e un atomo di alogeno. Un sottogruppo particolarmente studiato di composti secondo questa forma di realizzazione preferita è quello in cui Ri rappresenta idrogeno o metile, R4rappresenta idrogeno, R2e R3sono scelti indipendentemente tra idrogeno, metile, metossile e cloro. In this case, according to some preferred embodiments of the invention, R1, R2, R3 and R4 can be selected, each independently of the other, from H, an alkyl group with up to 6 carbon atoms, an alkoxy group with up to 6 carbon atoms and a halogen atom. A particularly studied subgroup of compounds according to this preferred embodiment is that in which R1 represents hydrogen or methyl, R4 represents hydrogen, R2 and R3 are independently selected from hydrogen, methyl, methoxyl and chlorine.
Appartiene a tale sottogruppo il composto isoindolo-chinossalinico di formula (2a): The isoindole-quinoxaline compound of formula (2a) belongs to this subgroup:
che si è dimostrato , nella sperimentazione condotta nel quadro della presente invenzione , quello con la migliore attività antitumorale in vitro tra i composti esaminati. Altri composti preferiti, che pure sono risultati attivi nell'inibire la crescita delle cellule tumorali utilizzate per la sperimentazione, sono quelli rappresentati dalle seguenti formule: which proved, in the experimentation carried out in the framework of the present invention, the one with the best antitumor activity in vitro among the compounds examined. Other preferred compounds, which have also been found to be active in inhibiting the growth of tumor cells used for experimentation, are those represented by the following formulas:
Nell’ulteriore caso specifico in cui, nella formula (1) che precede, R rappresenta H e R5rappresenta CN, i composti secondo l'invenzione sono descritti dalla formula generale (3): In the further specific case in which, in the formula (1) above, R represents H and R5 represents CN, the compounds according to the invention are described by the general formula (3):
Tra i composti di formula (3) un sottogruppo preferito è costituito d quelli in cui R1, R2, R3e R4sono scelti, indipendentemente uno dall'altro, tra H, un gruppo alchilico con fino a 6 atomi di carbonio, un gruppo alcossilico con fino a 6 atomi di carbonio e un atomo di alogeno. In modo ancora più specifico, un gruppo preferito di composti di formula (3) è quello in cui Ri rappresenta idrogeno o metile, R4rappresenta idrogeno, R2e R3sono scelti indipendentemente tra idrogeno, metile, metossile o cloro. Among the compounds of formula (3) a preferred subgroup consists of those in which R1, R2, R3 and R4 are selected, independently of each other, from H, an alkyl group with up to 6 carbon atoms, an alkoxy group with up to with 6 carbon atoms and one halogen atom. More specifically, a preferred group of compounds of formula (3) is that in which R1 represents hydrogen or methyl, R4 represents hydrogen, R2 and R3 are independently selected from hydrogen, methyl, methoxyl or chlorine.
Alla precedente definizione rispondono, in particolare, i derivati isoindolo-chinossalinici aventi una delle seguenti formule: In particular, the isoindole-quinoxaline derivatives having one of the following formulas respond to the previous definition:
Come già notato, i derivati isoindolo[2,1-a]chinossalinici dell'invenzione sono stati sintetizzati a partire da intermedi chiave, i 2-(2’-amminofenil)-1-ciano-isoindoli di formula (4) As already noted, the isoindole [2,1-a] quinoxaline derivatives of the invention have been synthesized starting from key intermediates, the 2- (2'-aminophenyl) -1-cyano-isoindoles of formula (4)
In letteratura, soltanto due derivati isoindolo[2,1-a]chinossalinici erano stati ottenuti come composti indesiderati nel corso del tentativo di sintetizzare il sistema benzoimidazo[2,1-a]isochinolinico (Dyker, G. et al. Oxidative Heterocyclization of 2-Alkylylbenzaldehydes with 1,2-Phenyiendiamine. Eur. J. Org. Chem. 2000, 1433-1441). In tale occasione, la reazione di 2-alchinil-arilaldeidi con o-fenilendiammina aveva dato come risultato una miscela inseparabile del sistema desiderato, benzoimidazo-isochinolinico, e delle isoindolo[2,1-a]chinossaline indesiderate. In the literature, only two isoindole [2,1-a] quinoxaline derivatives had been obtained as undesirable compounds in the course of an attempt to synthesize the benzoimidazo [2,1-a] isoquinoline system (Dyker, G. et al. Oxidative Heterocyclization of 2 -Alkylylbenzaldehydes with 1,2-Phenyiendiamine. Eur. J. Org. Chem. 2000, 1433-1441). On that occasion, the reaction of 2-alkynyl-arylaldehydes with o-phenylenediamine resulted in an inseparable mixture of the desired system, benzoimidazo-isoquinoline, and the unwanted isoindole [2,1-a] quinoxalines.
Secondo la procedura sintetica proposta per la presente invenzione, le isoindolo-chinossaline di formula (2) [cioè 2' o 2"] possono esere preparate per reazione dei corrispondenti intermedi isoindolici di formula (4) (2-(2'-amminofenil)-1-ciano-isoindoli) con acido acetico. In particolare, il riscaldamento a riflusso dei composti di formula (4) in cui R1-R4rappresentano i gruppi desiderati nel composto finale, in acido acetico, causa la reazione del gruppo amminico con il gruppo carbonitrile, dando origine alla formazione dell'anello chinossalinico. L’idrolisi in situ della funzione imminica formatasi risulta nei corrispondenti isoindolo-chinossalinoni di formula (2’) (isoindolo[2,1-a]chinossalin-6-(5H)oni) con rese eccellenti. According to the synthetic procedure proposed for the present invention, the isoindole-quinoxalines of formula (2) [i.e. 2 'or 2 "] can be prepared by reaction of the corresponding isoindole intermediates of formula (4) (2- (2'-aminophenyl) -1-cyano-isoindoles) with acetic acid. In particular, the reflux heating of the compounds of formula (4) in which R1-R4 represent the desired groups in the final compound, in acetic acid, causes the reaction of the amino group with the group carbonitrile, giving rise to the formation of the quinoxaline ring. The in situ hydrolysis of the imine function formed results in the corresponding isoindole-quinoxalinones of formula (2 ') (isoindole [2,1-a] quinoxalin-6- (5H) oni) with excellent yields.
Sempre a partire dagli intermedi di formula (4) recanti le funzioni R1-R4desiderate nel prodotto finale, i composti isoindolo-chinossalinici secondo l'invenzione che rientrano nella formula (3) (11-cianoisoindolo[2,1-a]chinossaline) possono essere preparati altrettanto semplicemente e con ottime rese per reazione di detti intermedi, sempre sotto riscaldamento (ad esempio a riflusso), con acido formico. Still starting from the intermediates of formula (4) bearing the R1-R4 functions desired in the final product, the isoindole-quinoxaline compounds according to the invention which fall within the formula (3) (11-cyanoisoindole [2,1-a] quinoxaline) can be prepared just as simply and with excellent yields by reaction of said intermediates, always under heating (for example under reflux), with formic acid.
La produzione degli intermedi di formula (4) è a sua volta possibile attraverso una sintesi di StrecKer tra le o-fenilendiammine di formula (5): The production of the intermediates of formula (4) is in turn possible through a synthesis of StrecKer among the o-phenylenediamines of formula (5):
in cui i sostituenti R1-R4rappresentano gli stessi gruppi della corrispondente formula (1), e la ftalodicarbossialdeide di formula (6): in which the substituents R1-R4 represent the same groups of the corresponding formula (1), and the phthaloodicarboxyaldehyde of formula (6):
(6) (6)
La reazione viene condotta in acqua e in presenza di cianuro di potassio e idrogenosolfito di sodio. Tale reazione, quando condotta con la 3-metil-o-1,2-fenilendiammina (composto di formula (5) con R1= Me e R2= R3= R4= H) ha dato, come prodotto di formula (4) oltre all'intermedio desiderato, anche l'isomero 1-ciano-2-(2’-ammino-6’-metil-fenil-isoindolo). Invece, nel caso della 3-metossi-o-1,2-fenilendiammina (composto di formula (5) con Ri = OMe e R2= R3= R4= H) è stato ottenuto soltanto l’intermedio di formula (4) con R2= OMe, a causa dell'aumentata reattività del gruppo amminico posto in para- rispetto al sostituente metossilico. The reaction is carried out in water and in the presence of potassium cyanide and sodium hydrogen sulfite. This reaction, when carried out with 3-methyl-o-1,2-phenylenediamine (compound of formula (5) with R1 = Me and R2 = R3 = R4 = H) gave, as product of formula (4) in addition to the desired intermediate, also the 1-cyano-2- (2'-amino-6'-methyl-phenyl-isoindole) isomer. Instead, in the case of 3-methoxy-o-1,2-phenylenediamine (compound of formula (5) with Ri = OMe and R2 = R3 = R4 = H) only the intermediate of formula (4) with R2 was obtained = OMe, due to the increased reactivity of the amino group placed in para- with respect to the methoxy substituent.
Costituisce pertanto ulteriore oggetto dell'invenzione un procedimento per la preparazione dei composti isoindolo-chinossalinici di formula (1), come sopra definita, comprendente la fase di far reagire un intermedio della formula generale (4), sopra illustrata, in cui R1, R2, R3e R4hanno il significato già enunciato, con acido acetico, oppure con acido formico, sotto riscaldamento. Therefore, a further object of the invention is a process for the preparation of isoindole-quinoxaline compounds of formula (1), as defined above, comprising the step of reacting an intermediate of the general formula (4), illustrated above, in which R1, R2 , R3and R4 have the meaning already stated, with acetic acid, or with formic acid, under heating.
Secondo alcune forme di realizzazione preferite dell’invenzione, quando il procedimento proposto è indirizzato alla produzione dei composti isoindolo-chinossalinici di formula (2'), esso comprende la fase di far reagire un intermedio della formula generale (4) con acido acetico a riflusso. Preferibilmente, al termine di detta reazione con acido acetico la miscela di reazione viene rapidamente raffreddata e il precipitato risultante, contenente il composto di formula (2’) desiderato, viene raccolto per filtrazione. According to some preferred embodiments of the invention, when the proposed process is directed to the production of the isoindole-quinoxaline compounds of formula (2 '), it comprises the step of making an intermediate of the general formula (4) react with acetic acid under reflux . Preferably, at the end of said reaction with acetic acid, the reaction mixture is rapidly cooled and the resulting precipitate, containing the desired compound of formula (2 '), is collected by filtration.
Secondo altre forme di realizzazione dell'invenzione, in cui il procedimento proposto è finalizzato alla produzione dei composti isoindolo-chinossalinici di formula (3), esso comprende la fase di far reagire un intermedio della formula generale (4) con acido formico a riflusso. Anche in questo caso, preferibilmente, al termine della reazione la miscela ottenuta viene rapidamente raffreddata e il precipitato risultante, contenente il composto di formula (3) desiderato, viene raccolto per filtrazione. According to other embodiments of the invention, in which the proposed process is aimed at the production of the isoindole-quinoxaline compounds of formula (3), it comprises the step of reacting an intermediate of the general formula (4) with reflux formic acid. Also in this case, preferably, at the end of the reaction the obtained mixture is rapidly cooled and the resulting precipitate, containing the desired compound of formula (3), is collected by filtration.
Secondo un suo diverso aspetto, l’invenzione ha anche ad oggetto l’uso di uno 0 più composti isoindolo-chinossalinici di formula generale (1), come sopra definita, per la produzione di un preparato farmaceutico, più in particolare un preparato ad attività antitumorale. in base ad alcune forme di realizzazione specifiche dell'invenzione, gli uno o più composti isoindolo-chinossalinici utilizzati sono composti di formula generale (2’) o (2"), mentre in base ad altre forme di realizzazione della stessa invenzione gli uno o più composti isoindolo-chinossalinici utilizzati sono composti di formula generale (3). According to a different aspect, the invention also relates to the use of one or more isoindole-quinoxaline compounds of general formula (1), as defined above, for the production of a pharmaceutical preparation, more particularly a preparation with active antitumor. according to some specific embodiments of the invention, the one or more isoindole-quinoxaline compounds used are compounds of general formula (2 ') or (2 "), while according to other embodiments of the same invention the one or most isoindole-quinoxaline compounds used are compounds of general formula (3).
Costituisce ulteriore oggetto della presente invenzione un preparato farmaceutico comprendente come principio attivo almeno uno dei composti isoindolo-chinossalinici di formula generale (1), assieme ad uno o più coadiuvanti e/o veicoli farmacologicamente accettabili. In maniera particolarmente preferita il preparato comprende come principio attivo un composto isoindolo-chinossalinico di formula generale generale (2’) o (2"). A further object of the present invention is a pharmaceutical preparation comprising as active principle at least one of the isoindole-quinoxaline compounds of general formula (1), together with one or more pharmacologically acceptable adjuvants and / or vehicles. Particularly preferably, the preparation comprises as an active ingredient an isoindole-quinoxaline compound of general general formula (2 ') or (2 ").
Come sarà illustrato dettagliatamente nel seguito, uno dei composti di formula (2) secondo l’invenzione ha mostrato un valore di Gl50(inibizione della crescita) a livello nanomolare nei confronti del 88% delle circa 60 linee cellulari tumorali umane in tutti i nove subpanel del test applicato: leucemia, tumore polmonare a cellule non piccole, tumore dei colon; tumore del SNC, melanoma; carcinoma ovarico, tumore renale, carcinoma prostatico, carcinoma mammario. As will be illustrated in detail below, one of the compounds of formula (2) according to the invention showed a value of Gl50 (growth inhibition) at the nanomolar level against 88% of the approximately 60 human tumor cell lines in all nine subpanels. of the test applied: leukemia, non-small cell lung cancer, colon cancer; CNS tumor, melanoma; ovarian cancer, kidney cancer, prostate cancer, breast cancer.
I preparati farmaceutici adatti per la somministrazione, a scopo terapeutico, degli agenti isoindolo-chinossalinici secondo l’invenzione possono essere formulati secondo tecniche convenzionali note agli esperti del settore, mediante l’uso di eccipienti e veicoli farmaceuticamente accettabili, in modo da ottenere preparati adatti per iniezione parenterale (endovenosa, intramuscolare o sottocutanea), per somministrazione orale in forma solida o liquida, per somministrazione transdermica, rettale, intravaginale o locale, ad esempio sulle mucose del cavo oronasale, e simili. Pharmaceutical preparations suitable for the administration, for therapeutic purposes, of the isoindole-quinoxaline agents according to the invention can be formulated according to conventional techniques known to those skilled in the art, through the use of pharmaceutically acceptable excipients and vehicles, so as to obtain suitable preparations for parenteral injection (intravenous, intramuscular or subcutaneous), for oral administration in solid or liquid form, for transdermal, rectal, intravaginal or local administration, for example on the mucous membranes of the oronasal cavity, and the like.
La percentuale di composto attivo nella composizione e il metodo per trattare i tumori possono essere variati in modo da ottenere un dosaggio ottimale. Il dosaggio da somministrare tiene normalmente in considerazione i seguenti fattori: la via di somministrazione, la durata del trattamento, il peso e le condizioni del paziente, l'efficacia del composto attivo e la responsività del paziente. The percentage of active compound in the composition and the method of treating tumors can be varied in order to obtain an optimal dosage. The dosage to be administered normally takes into account the following factors: the route of administration, the duration of treatment, the patient's weight and condition, the efficacy of the active compound and the patient's responsiveness.
Una volta determinato il corretto dosaggio dei principio attivo, la formulazione e la scelta di eccipienti e coadiuvanti adatti ad ogni singola esigenza possono essere fatte sulla base delle comuni conoscenze del settore. Once the correct dosage of the active ingredient has been determined, the formulation and choice of excipients and adjuvants suitable for each individual need can be made on the basis of common knowledge of the sector.
Le caratteristiche specifiche dell'invenzione, così come i vantaggi delia stessa sia in termini di sintesi dei derivati proposti che in termini della loro attività biologica e farmacologica, risulteranno più evidenti con riferimento alla descrizione dettagliata presentata a titolo meramente esemplificativo nel seguito, assieme ai risultati delle sperimentazioni effettuate su di essa. Alcuni risultati sperimentali sono anche illustrati nei disegni allegati, in cui: The specific characteristics of the invention, as well as the advantages of the same both in terms of synthesis of the proposed derivatives and in terms of their biological and pharmacological activity, will be more evident with reference to the detailed description presented merely by way of example below, together with the results of the experiments carried out on it. Some experimental results are also illustrated in the attached drawings, in which:
la Figura 1 mostra l’effetto del derivato isoindolo-chinossalinico (2a) secondo l’invenzione sull’aggregazione dei microtubulì: tubulina purificata è stata incubata in assenza o in presenza del composto in studio alle concentrazioni indicate, ed è stato rilevato l’andamento nel tempo della fluorescenza, dovuta all’incorporazione di un reporter fluorescente nei microtubuli nel corso della polimerizzazione della tubulina; e Figure 1 shows the effect of the isoindole-quinoxaline derivative (2a) according to the invention on the aggregation of microtubules: purified tubulin was incubated in the absence or in the presence of the compound under study at the indicated concentrations, and the trend was detected in the time of fluorescence, due to the incorporation of a fluorescent reporter into the microtubules during the polymerization of tubulin; And
la Figura 2 mostra l’effetto di concentrazioni crescenti del derivato isoindolo-chinossalinico (2a) secondo l'invenzione sul rilassamento del DNA del plasmide pBR322 da parte di topoisomerasi I umana, rilevato per mezzo di elettroforesi su gel di agarosio con (sezione A) o senza (sezione B) bromuro di etidio. (Sc: supercoiled, Rel: relaxed, Nick: nicked). Figure 2 shows the effect of increasing concentrations of the isoindole-quinoxaline derivative (2a) according to the invention on the relaxation of the DNA of the plasmid pBR322 by human topoisomerase I, detected by means of agarose gel electrophoresis with (section A) or without (section B) ethidium bromide. (Sc: supercoiled, Rel: relaxed, Nick: nicked).
Per quanto riguarda la sintesi, come già notato, tutti i derivati isoindolo-chìnossalinici descrìtti nel seguito sono stati ottenuti a partire da intermedi di formula (4), i quali a loro volta erano ottenibili a partire dai due composti di partenza delle formule (5), o-fenilendiammina opportunamente sostituita, e (6) ftalodicarbossialdeide. Nella sperimentazione chimica illustrata di seguito tutti i punti di fusione sono stati presi con un apparato capillare Buchi-Tottoli; gli spettri IR sono stati determinati in bromoformio con uno spettrofotometro Jasco FT/IR 5300; gli spettri 1H and<13>C NMR sono stati misurati a 200 e 50,3 MHz rispettivamente, impiegando uno spettrometro Bruker serie AC 200 MHz (TMS come riferimento interno). La cromatografia su colonna è stata eseguita con gel di silice Merck 230-400 Mesh ASTM o con apparato Sepacore Buchi. Le analisi elementari erano nel campo ±0.4% dei valori teorici. As regards the synthesis, as already noted, all the isoindole-chinoxaline derivatives described below were obtained starting from intermediates of formula (4), which in turn were obtainable starting from the two starting compounds of the formulas (5 ), suitably substituted o-phenylenediamine, and (6) phthalodicarboxyaldehyde. In the chemical experimentation illustrated below, all the melting points were taken with a Buchi-Tottoli capillary apparatus; the IR spectra were determined in bromoform with a Jasco FT / IR 5300 spectrophotometer; the 1H and <13> C NMR spectra were measured at 200 and 50.3 MHz respectively, using a Bruker series AC 200 MHz spectrometer (TMS as internal reference). Column chromatography was performed with Merck 230-400 Mesh ASTM silica gel or Sepacore Buchi apparatus. The elementary analyzes were in the range ± 0.4% of the theoretical values.
ESEMPIO 1 EXAMPLE 1
lsoindolo[2.1 -a]chinossalin-6-(5H)one (2b) Preparazione dell'intermedio di formula (4): 1-ciano-2-(2<,>-amminofenil)-isoindolo lsoindole [2.1 -a] quinoxalin-6- (5H) one (2b) Preparation of the intermediate of formula (4): 1-cyano-2- (2 <,> - aminophenyl) -isoindole
Ad una soluzione di 1,56 g (0,015 mol) di idrogenosolfito di sodio in acqua (38 mi) è stata aggiunta ftalodicarbossialdeide (6) (2 g, 0,015 mol). La miscela è stata tenuta sotto agitazione fino a dissoluzione del solido ed è stata poi aggiunta la o-fenilendiammina di formula (5), in cui tutti i sostituenti R1-R4rappresentano idrogeno (0,015 mol), e la reazione è stata tenuta sotto riscaldamento su un bagno di vapore acqueo per 30 min. Quindi è stato aggiunto KCN (3,39 g, 0,052 mol) in acqua (8,0 mi) e la miscela è stata riscaldata per ulteriori 90 min. Phthalodicarboxyaldehyde (6) (2 g, 0.015 mol) was added to a solution of 1.56 g (0.015 mol) of sodium hydrogen sulfite in water (38 ml). The mixture was kept under stirring until the solid dissolved and then the o-phenylenediamine of formula (5) was added, in which all the substituents R1-R4 represent hydrogen (0.015 mol), and the reaction was kept under heating on a water vapor bath for 30 min. Then KCN (3.39 g, 0.052 mol) in water (8.0 ml) was added and the mixture was heated for an additional 90 min.
La miscela di reazione è stata poi raffreddata e il solido formato è stato filtrato e purificato per cromatografia. The reaction mixture was then cooled and the solid formed was filtered and purified by chromatography.
1-ciano-2-(2’-amminofenil)-isoindolo 1-cyano-2- (2'-aminophenyl) -isoindole
Il composto è stato purificato per flash-cromatografia utilizzando diclorometano come eiuente a dare un solido: resa 65%; p.f. 150-151°C; The compound was purified by flash chromatography using dichloromethane as an euent to give a solid: yield 65%; m.p. 150-151 ° C;
IR 3473, 3363 (NH2), 2202 (CN) cm<-1>;<1>H NMR (DMSO-d6) δ 5.19 (s, 2H, NH2), 6.70 (t, 1H, J=7.2 Hz, H-5’), 6.94 (d, 1H, J=7.2 Hz, H-3’), 7.15-7.19 (m, 3H, H-4’, H-5 e H-6’), 7.31 (t, 1H, J=8.2 Hz, H-6), 7.66 (d, 1H, J=8.2 Hz, H-4), 7.78 (d, 1H, J=8.2 Hz, H-7), 7.86 (s, 1H, H-3);<13>C NMR (DMSO-de) δ 93.9 (s), 114.0 (s), 115.8 (d), 116.1 (d), 117.5 (d), 121.5 (d), 122.2 (d), 122.4 (d, C-7), 122.5 (s), 124.1 (s), 125.5 (d), 127.8 (d), 130.4 (d), 131.2 (s), 144.1 (s). Anal. (C15H1N3) C, H, N. IR 3473, 3363 (NH2), 2202 (CN) cm <-1>; <1> H NMR (DMSO-d6) δ 5.19 (s, 2H, NH2), 6.70 (t, 1H, J = 7.2 Hz, H -5 '), 6.94 (d, 1H, J = 7.2 Hz, H-3'), 7.15-7.19 (m, 3H, H-4 ', H-5 and H-6'), 7.31 (t, 1H , J = 8.2 Hz, H-6), 7.66 (d, 1H, J = 8.2 Hz, H-4), 7.78 (d, 1H, J = 8.2 Hz, H-7), 7.86 (s, 1H, H -3); <13> C NMR (DMSO-de) δ 93.9 (s), 114.0 (s), 115.8 (d), 116.1 (d), 117.5 (d), 121.5 (d), 122.2 (d), 122.4 (d, C-7), 122.5 (s), 124.1 (s), 125.5 (d), 127.8 (d), 130.4 (d), 131.2 (s), 144.1 (s). Anal. (C15H1N3) C, H, N.
Isoindolo[2,1 -a]chinossalin-6-(5H)one (2b) Isoindole [2,1 -a] quinoxalin-6- (5H) one (2b)
L’intermedio 1-ciano-2-(2’-amminofenil)-isoindolo ottenuto dalla fase precedente (3 mmoli) è stato disciolto in acido acetico (10 ml) e tenuto a riflusso per 30 min. La miscela di reazione è stata versata in acqua ghiacciata (150 ml) e il precipitato risultante è stato raccolto per filtrazione e cristallizzato da etanolo. The intermediate 1-cyano-2- (2'-aminophenyl) -isoindole obtained from the previous phase (3 mmoles) was dissolved in acetic acid (10 ml) and refluxed for 30 min. The reaction mixture was poured into ice water (150 ml) and the resulting precipitate was collected by filtration and crystallized from ethanol.
Resa 99%; p.f. 255-256°C; Yield 99%; m.p. 255-256 ° C;
IR; 3390, 3203 (NH), 1685 (CO) cm<-1>;<1>H NMR (DMSO-d6) δ 7.75 (t, 1H, J=7.0 , 1.6 Hz, H-2), 7.78-7.91 (m, 2H, H-3 e l-M), 7.93 (s, 1H, H-11), 7.96 (t, 1H, J=7.8 Hz, H-9), 8,00 (t, 1H, J=7.8 Hz, H-8), 8.27 (dd, 1H, J=7.0, 1.6 Hz, H-1), 8.34 (d, 1H, J=7.8 Hz, H-10), 8.84 (d, 1H, J=7.8 Hz, H-7), 12.35 (bs, 1H, NH);<13>C NMR (DMSO-de) δ 124.1 (d), 127.0 (d), 127.5 (d), 127.8 (d), 127.9 (d), 128.9 (d), 130,9 (d), 131.4 (d), 133.1 (s), 133.4 (d), 135.3 (s), 138.9 (s), 141.0 (s), 143.9 (s), 161.7 (s). Anai. (C15H10N2O) C, H, N. IR; 3390, 3203 (NH), 1685 (CO) cm <-1>; <1> H NMR (DMSO-d6) δ 7.75 (t, 1H, J = 7.0, 1.6 Hz, H-2), 7.78-7.91 ( m, 2H, H-3 and l-M), 7.93 (s, 1H, H-11), 7.96 (t, 1H, J = 7.8 Hz, H-9), 8.00 (t, 1H, J = 7.8 Hz , H-8), 8.27 (dd, 1H, J = 7.0, 1.6 Hz, H-1), 8.34 (d, 1H, J = 7.8 Hz, H-10), 8.84 (d, 1H, J = 7.8 Hz , H-7), 12.35 (bs, 1H, NH); <13> C NMR (DMSO-de) δ 124.1 (d), 127.0 (d), 127.5 (d), 127.8 (d), 127.9 (d) , 128.9 (d), 130.9 (d), 131.4 (d), 133.1 (s), 133.4 (d), 135.3 (s), 138.9 (s), 141.0 (s), 143.9 (s), 161.7 ( s). Anai. (C15H10N2O) C, H, N.
ESEMPIO 2 EXAMPLE 2
4-metil-isoindolo[2, 1 -a]chinossalin-6-(5H)one (2c) La preparazione dell'ntermedio di formula (4): 1-ciano-2-(2’-ammino-3’-metil-fenil)-isoindolo è stata eseguita come nell'Esempio 1 , con la differenza che la o-fenilendiammina di formula (5), era una metilfenilendiammina (Ri = Me). 4-methyl-isoindole [2, 1 -a] quinoxalin-6- (5H) one (2c) The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-3'-methyl -phenyl) -isoindole was carried out as in Example 1, with the difference that the o-phenylenediamine of formula (5) was a methylphenylenediamine (Ri = Me).
1 -ciano-2-(2’-ammino-3’-metil-fenil)-isoindolo (4b) 1-cyan-2- (2'-amino-3'-methyl-phenyl) -isoindole (4b)
Il composto è stato purificato con l’apparato Sepacore Buchi utilizzando diclorometanoicicloesano (7:3) come eluente a dare il composto 4b: resa 40%; p.f. 62-63°C; The compound was purified with the Sepacore Buchi apparatus using dichlorometanoicyclohexane (7: 3) as eluent to give compound 4b: yield 40%; m.p. 62-63 ° C;
IR 3481, 3388 (NH2), 2200 (CN) cm<'1>; NMR (CDCI3) δ 2.25 (s, 3H, CH3), 3.55 (s, 2H, NH2), 6.80 (t, 1H, J=7.6 Hz, H-5'), 7.10 (d, 1H, J-7.6 Hz, H-6’), 7.14 (d, 1H, J=7.6 Hz, H-4’), 7.23 (t, 1H, J=8.1 Hz, H5), 7.31 (t, 1H, J=8.1 Hz, H-6), 7.44 (s, 1H, H-3), 7.69 (d, 1H, J=8.1 Hz, H-4), 7.73 (d, 1H, J=8.1 Hz, H-7);<13>C NMR (CDCI3) δ 17.6 (q), 92.2 (s), 113.0 (s), 117.9 (d), 118.3 (d), 120.5 (d), 120.9 (d), 123.2 (d), 123.8 (s), 124.1 (s), 124.6 (s), 125.5 <d), 125.9 (d), 131.8 (s), 131.9 (d), 140.5 (s). Anal. (CI6HI3N3) C, H, N. IR 3481, 3388 (NH2), 2200 (CN) cm <'1>; NMR (CDCI3) δ 2.25 (s, 3H, CH3), 3.55 (s, 2H, NH2), 6.80 (t, 1H, J = 7.6 Hz, H-5 '), 7.10 (d, 1H, J-7.6 Hz , H-6 '), 7.14 (d, 1H, J = 7.6 Hz, H-4'), 7.23 (t, 1H, J = 8.1 Hz, H5), 7.31 (t, 1H, J = 8.1 Hz, H -6), 7.44 (s, 1H, H-3), 7.69 (d, 1H, J = 8.1 Hz, H-4), 7.73 (d, 1H, J = 8.1 Hz, H-7); <13> C NMR (CDCI3) δ 17.6 (q), 92.2 (s), 113.0 (s), 117.9 (d), 118.3 (d), 120.5 (d), 120.9 (d), 123.2 (d), 123.8 (s) , 124.1 (s), 124.6 (s), 125.5 <d), 125.9 (d), 131.8 (s), 131.9 (d), 140.5 (s). Anal. (CI6HI3N3) C, H, N.
4-metil-isoindolo[2,1-a]chinossalin-6-(5H)one (2c) 4-methyl-isoindole [2,1-a] quinoxalin-6- (5H) one (2c)
L’intermedio ottenuto dalla fase precedente è stato trattato con acido acetico come nell’Esempio 1. The intermediate obtained from the previous phase was treated with acetic acid as in Example 1.
Resa 98%; p.f. 338-339°C; Yield 98%; m.p. 338-339 ° C;
IR: 3360, 3284 (NH), 1632 (CO) cm<1>;<1>H NMR (DMSO-d6) δ 2.62 (s, 3H, CH3), 7.32-7.40 (m, 2H, H-9 e H-3), 7.45 (t, 1H, J=6.8 Hz, H-2), 7.51 (t, 1H, J=8.8 Hz, H-8), 7.96 (d, 1H, J=8.8 Hz, H-10), 8.27 (d, 1H, J=6.8 Hz, H-1), 8.47 (d, 1H, J=8.8 Hz, H-7), 9.24 (s, 1H, H-11), 12.53 (bs, IH, NH);<13>C NMR (DMSO-d6) δ 17.8 (q), 104.8(s), 114.8 (d), 117.3 (d), 119.2 (d), 121.5 (d), 122.3 (s), 125.0 (d), 125.1 (s), 125.2 (d), 125.3 (s), 127.1 (d), 127.3 (s x2), 130.0 (d), 147.0 (s). Anal. (C16H12N20) C, H, N. IR: 3360, 3284 (NH), 1632 (CO) cm <1>; <1> H NMR (DMSO-d6) δ 2.62 (s, 3H, CH3), 7.32-7.40 (m, 2H, H-9 and H-3), 7.45 (t, 1H, J = 6.8 Hz, H-2), 7.51 (t, 1H, J = 8.8 Hz, H-8), 7.96 (d, 1H, J = 8.8 Hz, H- 10), 8.27 (d, 1H, J = 6.8 Hz, H-1), 8.47 (d, 1H, J = 8.8 Hz, H-7), 9.24 (s, 1H, H-11), 12.53 (bs, 1H, NH); <13> C NMR (DMSO-d6) δ 17.8 (q), 104.8 (s), 114.8 (d), 117.3 (d), 119.2 (d), 121.5 (d), 122.3 (s) , 125.0 (d), 125.1 (s), 125.2 (d), 125.3 (s), 127.1 (d), 127.3 (s x2), 130.0 (d), 147.0 (s). Anal. (C16H12N20) C, H, N.
ESEMPIO 3 EXAMPLE 3
3-metossi-isoindolo[2.1-a]chinossalin-6-(5H)one (2a) La preparazione dell’intermedio di formula (4): 1-ciano-2-(2’-ammino-4’-metossi-fenil)-isoindolo è stata eseguita come nell’Esempio 1, con la differenza che la o-fenilendiammina di formula (5), era una metossi-fenilendiammina (R2= OMe). 3-methoxy-isoindole [2.1-a] quinoxalin-6- (5H) one (2a) The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-4'-methoxy-phenyl ) -isoindole was carried out as in Example 1, with the difference that the o-phenylenediamine of formula (5), was a methoxy-phenylenediamine (R2 = OMe).
1 -cìano-2-(2’-ammino-4’-metossi-fenil)-isoindolo (4c) 1-cyano-2- (2'-amino-4'-methoxy-phenyl) -isoindole (4c)
il composto è stato purificato per flash-cromatografia utilizzando diclorometano come eluente a dare un solido: resa 95%; p.f. 116-117<º>C; the compound was purified by flash chromatography using dichloromethane as eluent to give a solid: yield 95%; m.p. 116-117 <º> C;
IR 3465, 3375 (NH2), 2198 (CN) cm<-1>; NMR (CDCI3) 6 3.64 (bs, 2H, NH2), 3.80 (s, 3H, CH3), 6.38 (bs, 1H, H-3’), 6.42 (d, 1H, J=7.9 Hz, H-5'), 7.12 (d, 1H, J=7.9 Hz, H-6’), 7.14 (t, 1H, J=8.6 Hz, H-5), 7.29 (t, 1H, J=8.6 Hz, H-6), 7.41 (s, 1H, H-3), 7.66 (d, 1H, J=8.6 Hz, H-4), 7.70 (d, 1H, J=8.6 Hz, H-7);<13>C NMR (CDCI3) δ 55.4 (q), 95.4 (s), 101.4 (d), 104.5 (d), 113.9 (s), 117.5 (s), 118.2 (d), 120.8 (dx2), 123.1 (d), 124.5 (s), 125.8 (d), 128.7 (d), 131.7 (s), 143.3 (s), 161.5 (s). Anal. (CI6H13N30) C, H, N. IR 3465, 3375 (NH2), 2198 (CN) cm <-1>; NMR (CDCI3) 6 3.64 (bs, 2H, NH2), 3.80 (s, 3H, CH3), 6.38 (bs, 1H, H-3 '), 6.42 (d, 1H, J = 7.9 Hz, H-5' ), 7.12 (d, 1H, J = 7.9 Hz, H-6 '), 7.14 (t, 1H, J = 8.6 Hz, H-5), 7.29 (t, 1H, J = 8.6 Hz, H-6) , 7.41 (s, 1H, H-3), 7.66 (d, 1H, J = 8.6 Hz, H-4), 7.70 (d, 1H, J = 8.6 Hz, H-7); <13> C NMR ( CDCI3) δ 55.4 (q), 95.4 (s), 101.4 (d), 104.5 (d), 113.9 (s), 117.5 (s), 118.2 (d), 120.8 (dx2), 123.1 (d), 124.5 ( s), 125.8 (d), 128.7 (d), 131.7 (s), 143.3 (s), 161.5 (s). Anal. (CI6H13N30) C, H, N.
3-metossi-isoindolo[2,1-a]chinossalin-6-(5H)one (2a) L'intermedio ottenuto dalla fase precedente è stato trattato con acido acetico come nell’Esempio 1. 3-methoxy-isoindole [2,1-a] quinoxalin-6- (5H) one (2a) The intermediate obtained from the previous phase was treated with acetic acid as in Example 1.
Resa 75%; p.f. 293-294<º>C; Yield 75%; m.p. 293-294 <º> C;
IR: 3363, 3280 (NH), 1635 (CO) cm<'1>;<1>H NMR (DMSO-d6) δ 3.84 (3H, s, CH3), 7.04 (s, 1H, H-4), 7.05 (d, 1H, J=9.8 Hz, H-2), 7.40 (t, 1H, J=7.8 Hz, H-9), 7.51 (t, 1H, J=7.8 Hz, H-8), 7.95 (d, 1H, J=7.8 Hz, H-10), 8.36 (d, 1H, J=9.8 Hz, H-1), 8.51 (d, 1H, J=7.8 Hz, H-7), 9.18 (s, 1H, H-11), 13.50 (bs, 1H, NH);<13>C NMR (DMSO-ds) δ 55.7 (q), 101.5 (d), 104.6 (s), 112.7 (d), 115.7 (d), 116.2 (s), 118.2 (d), 119.1 (d), 120.9 (d), 124.3 (d), 124.4 (s), 126.2 (d), 126.6 (s), 128.5 (s), 147.1 (s), 158.9 (s). Anal. (C16Hi2N202) C, H, N. IR: 3363, 3280 (NH), 1635 (CO) cm <'1>; <1> H NMR (DMSO-d6) δ 3.84 (3H, s, CH3), 7.04 (s, 1H, H-4), 7.05 (d, 1H, J = 9.8 Hz, H-2), 7.40 (t, 1H, J = 7.8 Hz, H-9), 7.51 (t, 1H, J = 7.8 Hz, H-8), 7.95 ( d, 1H, J = 7.8 Hz, H-10), 8.36 (d, 1H, J = 9.8 Hz, H-1), 8.51 (d, 1H, J = 7.8 Hz, H-7), 9.18 (s, 1H, H-11), 13.50 (bs, 1H, NH); <13> C NMR (DMSO-ds) δ 55.7 (q), 101.5 (d), 104.6 (s), 112.7 (d), 115.7 (d ), 116.2 (s), 118.2 (d), 119.1 (d), 120.9 (d), 124.3 (d), 124.4 (s), 126.2 (d), 126.6 (s), 128.5 (s), 147.1 (s ), 158.9 (s). Anal. (C16Hi2N202) C, H, N.
ESEMPIO 4 EXAMPLE 4
2.3-dimetil-isoindolo[2.1-a]chinossalin-6-(5H)one (2d) La preparazione dell’intermedio di formula (4): 1-ciano-2-(2’ammino-4',5’-dimetil-fenil)-isoindolo è stata eseguita come nell’Esempio 1, con la differenza che la o-fenilendiammina di formula (5), era una dimetil-fenilendiammina (R2= R3= Me). 2.3-dimethyl-isoindole [2.1-a] quinoxalin-6- (5H) one (2d) The preparation of the intermediate of formula (4): 1-cyano-2- (2'amino-4 ', 5'-dimethyl -phenyl) -isoindole was carried out as in Example 1, with the difference that the o-phenylenediamine of formula (5), was a dimethyl-phenylenediamine (R2 = R3 = Me).
1 -ciano-2-(2’-ammino-4<,>,5<,>-dimetilfenil)-isoindolo (4d) 1-cyan-2- (2'-amino-4 <,>, 5 <,> - dimethylphenyl) -isoindole (4d)
Il composto è stato purificato per flash-cromatografia utilizzando diclorometano come eluente a dare un solido: resa 85%; p.f. 119-120°C; The compound was purified by flash chromatography using dichloromethane as eluent to give a solid: yield 85%; m.p. 119-120 ° C;
IR 3465, 3375 (NH2), 2202 (CN) cm<-1>; NMR (CDCI3) δ 2.17 (s, 3H, CH3), 2.22 (s, 3H, CH3), 3.44 (s, 2H, NH2), 6.64 (s, 1H, H-3’), 6.95 (s, 1H, H-6'), 7.12 (t, 1H, J=7.6 Hz, H-5), 7.27 (t, 1H, J=7.6 Hz, H-6), 7.38 (s, 1H, H-3), 7.64 (d, 1H, J=7.6 Hz, H-4), 7.68 (d, 1H, J=7.6 Hz, H-7);<13>C NMR (CDCI3) δ 18.6 (q), 19.7 (q), 95.1 (s), 114.0 (s), 118.1 (d), 118.2 (d), 120.6 (d), 120.9 (d), 121.8 (s), 123.1 (d), 124.5 (s), 125.8 (d), 127.0 (s), 128.2 (d), 131.7 (s), 139.6 (s), 139.7 (s). Anal. (C17H15N3): C, H, N. IR 3465, 3375 (NH2), 2202 (CN) cm <-1>; NMR (CDCI3) δ 2.17 (s, 3H, CH3), 2.22 (s, 3H, CH3), 3.44 (s, 2H, NH2), 6.64 (s, 1H, H-3 '), 6.95 (s, 1H, H-6 '), 7.12 (t, 1H, J = 7.6 Hz, H-5), 7.27 (t, 1H, J = 7.6 Hz, H-6), 7.38 (s, 1H, H-3), 7.64 (d, 1H, J = 7.6 Hz, H-4), 7.68 (d, 1H, J = 7.6 Hz, H-7); <13> C NMR (CDCI3) δ 18.6 (q), 19.7 (q), 95.1 (s), 114.0 (s), 118.1 (d), 118.2 (d), 120.6 (d), 120.9 (d), 121.8 (s), 123.1 (d), 124.5 (s), 125.8 (d), 127.0 (s), 128.2 (d), 131.7 (s), 139.6 (s), 139.7 (s). Anal. (C17H15N3): C, H, N.
2,3-dimetil-isoindolo[2,1 -a]chinossalin-6-(5H)one (2d) 2,3-dimethyl-isoindole [2,1 -a] quinoxalin-6- (5H) one (2d)
L'intermedio ottenuto dalla fase precedente è stato trattato con acido acetico come nell’Esempio 1. The intermediate obtained from the previous phase was treated with acetic acid as in Example 1.
Resa 96%; p.f. 370-371 °C; Yield 96%; m.p. 370-371 ° C;
IR: 3365, 3286 (NH), 1631 (CO) cm<'1>;<1>H NMR (DMSO-d6) δ 2.30 (3H, s, CH3), 2.35 (3H, s, CH3), 7.31 (s, 1H, H-4), 7.46 (t, 1H, J=7.8 Hz, H-9), 7.57 (t, 1H, J=7.8 Hz, H-8), 8.00 (d, 1H, J=7.8 Hz, H-10), 8.09 (s, 1H, H-1), 8.41 (d, 1H, J=7.8 Hz, H-7), 9.02 (s, 1H, H-11), 13.14 (bs, 1H, NH);<13>C NMR (DMSO-d6) δ 19.5 (q), 19.6 (q), 116.7 (d), 117.4 (d), 118.7 (s), 119.1 (d), 119.6 (d), 121.0 (s), 122.0 (d), 125.3 (s), 125.4 (d), 125.8 (s), 127.6 (d), 127.9 (s), 135.9 (s), 139.0 (s), 147.2 (s). Anal. (C17H14N2O)C,H,N. IR: 3365, 3286 (NH), 1631 (CO) cm <'1>; <1> H NMR (DMSO-d6) δ 2.30 (3H, s, CH3), 2.35 (3H, s, CH3), 7.31 ( s, 1H, H-4), 7.46 (t, 1H, J = 7.8 Hz, H-9), 7.57 (t, 1H, J = 7.8 Hz, H-8), 8.00 (d, 1H, J = 7.8 Hz, H-10), 8.09 (s, 1H, H-1), 8.41 (d, 1H, J = 7.8 Hz, H-7), 9.02 (s, 1H, H-11), 13.14 (bs, 1H , NH); <13> C NMR (DMSO-d6) δ 19.5 (q), 19.6 (q), 116.7 (d), 117.4 (d), 118.7 (s), 119.1 (d), 119.6 (d), 121.0 (s), 122.0 (d), 125.3 (s), 125.4 (d), 125.8 (s), 127.6 (d), 127.9 (s), 135.9 (s), 139.0 (s), 147.2 (s). Anal. (C17H14N2O) C, H, N.
ESEMPIO 5 EXAMPLE 5
2.3-dicloro-isoindolo[2.1-a]chinossalin-6-(5H)one (2e) La preparazione dell’intermedio di formula (4): 1-ciano-2-(2’-ammino-4'5’-dicloro-fenil)-isoindolo è stata eseguita come nell'Esempio 1, con la differenza che la o-fenilendiammina di formula (5), era una dicloro-fenilendiammina (R2= R3= CI), e che in questo caso tale reagente è stato prima disciolto in DMF (15ml) e poi aggiunto alla miscela di reazione. 2.3-dichloro-isoindole [2.1-a] quinoxalin-6- (5H) one (2e) The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-4'5'-dichloro -phenyl) -isoindole was carried out as in Example 1, with the difference that the o-phenylenediamine of formula (5), was a dichloro-phenylenediamine (R2 = R3 = CI), and that in this case this reagent was first dissolved in DMF (15ml) and then added to the reaction mixture.
1 -ciano-2- -ammino-4,5’-diclorofenil)soindolo (4e) 1-cyan-2- -amino-4,5'-dichlorophenyl) soindole (4e)
Il composto è stato purificato per flash-cromatografia utilizzando diclorometano come eluente a dare un solido: resa 77%; p.f. 71-72<º>C; The compound was purified by flash chromatography using dichloromethane as eluent to give a solid: yield 77%; m.p. 71-72 <º> C;
IR 3394, 3340 (NH2), 2200 (CN) cm<-1>;<1>H NMR (DMSO-d6) δ 5.75 (2H, s, NH2), 7.12 (1 H, s, H-3'), 7.16 (1H, t, J=8.4 Hz, H-5), 7.32 (1 H, t, J=8.4 Hz, H-6), 7.58 (1H, s, H-6’), 7.66 (1H, d, J=8.4 Hz, H-4), 7.78 (1H, d, J=8.4 Hz, H-7), 7.91 (1H, s, H-3);<13>C NMR (DMSO-d6) δ 94.0 (s), 113.8 (s), 115.8 (s), 116.2 (d), 117.5 (d), 121.6 (d), 121.7 (s), 122.4 (d), 122.6 (d), 124.2 (e), 125.8 (d), 129.6 (d), 131.2 (s), 132.9 (s), 144.7 (s). Anal. (C15HeCI2N3) C, H, N. IR 3394, 3340 (NH2), 2200 (CN) cm <-1>; <1> H NMR (DMSO-d6) δ 5.75 (2H, s, NH2), 7.12 (1 H, s, H-3 ') , 7.16 (1H, t, J = 8.4 Hz, H-5), 7.32 (1 H, t, J = 8.4 Hz, H-6), 7.58 (1H, s, H-6 '), 7.66 (1H, d, J = 8.4 Hz, H-4), 7.78 (1H, d, J = 8.4 Hz, H-7), 7.91 (1H, s, H-3); <13> C NMR (DMSO-d6) δ 94.0 (s), 113.8 (s), 115.8 (s), 116.2 (d), 117.5 (d), 121.6 (d), 121.7 (s), 122.4 (d), 122.6 (d), 124.2 (e), 125.8 (d), 129.6 (d), 131.2 (s), 132.9 (s), 144.7 (s). Anal. (C15HeCI2N3) C, H, N.
2,3-dicloro-isoindolo[2,1-a]chinossalin-6-(5H)one (2e) 2,3-dichloro-isoindole [2,1-a] quinoxalin-6- (5H) one (2e)
L’intermedio ottenuto dalla fase precedente è stato trattato con acido acetico come nell'Esempio 1. The intermediate obtained from the previous phase was treated with acetic acid as in Example 1.
Resa 87%; p.f. 340-343°C; Yield 87%; m.p. 340-343 ° C;
IR: 3350, 3302 (NH), 1645 (CO) cm<-1>;<1>H NMR (DMSO-d6) δ 7.43 (t, 1H, J=7.8 Hz, H-9), 7.54 (t, 1H, J=7.8 Hz, H-8), 7.86 (s, 1H, H-4), 7.95 (d, 1H, J=7.8 Hz, H-10), 8.52 (d, 1H, J=7.8 Hz, H-7), 8.85 (s, 1H, H-1), 9.32 (s, 1H, H-11), 12.46 (bs, 1H, NH);<13>C NMR (DMSO-d6) δ 105.1 (s), 117.1 (d), 118.7 (d), 119.2 (d), 119.4 (d), 121.1 (d), 122.0 (s), 124.7 (s), 124.8 (d), 126.5 (s), 126.9 (s), 127.0 (d), 127.2 (s), 130.3 (s), 147.0 (s). Anal. (C15H0CI2N2O) C, H, N. IR: 3350, 3302 (NH), 1645 (CO) cm <-1>; <1> H NMR (DMSO-d6) δ 7.43 (t, 1H, J = 7.8 Hz, H-9), 7.54 (t, 1H, J = 7.8 Hz, H-8), 7.86 (s, 1H, H-4), 7.95 (d, 1H, J = 7.8 Hz, H-10), 8.52 (d, 1H, J = 7.8 Hz, H-7), 8.85 (s, 1H, H-1), 9.32 (s, 1H, H-11), 12.46 (bs, 1H, NH); <13> C NMR (DMSO-d6) δ 105.1 (s ), 117.1 (d), 118.7 (d), 119.2 (d), 119.4 (d), 121.1 (d), 122.0 (s), 124.7 (s), 124.8 (d), 126.5 (s), 126.9 (s ), 127.0 (d), 127.2 (s), 130.3 (s), 147.0 (s). Anal. (C15H0CI2N2O) C, H, N.
ESEMPIO 6 EXAMPLE 6
11-cianoisoindolo[2.1-a]chinossalina (3a) L’intermedio 1-ciano-2-{2’-amminofenil)-isoindolo ottenuto come nella prima parte dell'Esempio 1 (3 mmoli) è stato disciolto in acido formico al 99% (10 mi) e tenuto a riflusso fino alla completa scomparsa del materiale di partenza, cioè per un tempo totale di 15 h. La miscela di reazione è stata versata in acqua ghiacciata (150 mi) e il precipitato risultante è stato raccolto per filtrazione. Il solido ottenuto è stato purificato per flash-cromatografia utilizzando diclorometano a dare il prodotto del titolo, solido. 11-cyanoisoindole [2.1-a] quinoxaline (3a) The intermediate 1-cyano-2- {2'-aminophenyl) -isoindole obtained as in the first part of Example 1 (3 mmol) was dissolved in 99 formic acid % (10 ml) and refluxed until the complete disappearance of the starting material, ie for a total time of 15 h. The reaction mixture was poured into ice water (150ml) and the resulting precipitate was collected by filtration. The solid obtained was purified by flash chromatography using dichloromethane to give the title product, solid.
Resa 70%; p.f. 237-238°C; Yield 70%; m.p. 237-238 ° C;
IR: 2192 (CN) cm<'1>;<1>H NMR (DMSO-d6) δ 7.55 (t, 1H, J=7.2 Hz, H-8), 7.68 (t, 1H, J=7.2 Hz, H-9), 7.81 (d, 1H, J=7.2 Hz, H-7), 7.87 (t, 1H, J=8.0 Hz, H-2), 7.90 (t, 1H, J=8.0 Hz, H-3), 8.20 (d, 1H, J=7.2 Hz, H-10), 8.57 (d, 1H, J=8.0 Hz, H-1), 9.06 (d, 1H, J=8.0 Hz, H-4) 9.79 (s, 1H, H-6);<13>C NMR (DMSO-d6) δ 89.4(s), 115.4 (s), 115.7 (d), 116.9 (d), 119.8 (s), 120.3 (d), 121.7 (s), 124.7 (d), 127.2 (s), 128.4 (d), 128.5 (d), 129.0 (d), 139.7 (d), 132.2 (s), 138.5 (s), 143.3 (d), Anal. (C16H9N3) C, H, N. IR: 2192 (CN) cm <'1>; <1> H NMR (DMSO-d6) δ 7.55 (t, 1H, J = 7.2 Hz, H-8), 7.68 (t, 1H, J = 7.2 Hz, H-9), 7.81 (d, 1H, J = 7.2 Hz, H-7), 7.87 (t, 1H, J = 8.0 Hz, H-2), 7.90 (t, 1H, J = 8.0 Hz, H- 3), 8.20 (d, 1H, J = 7.2 Hz, H-10), 8.57 (d, 1H, J = 8.0 Hz, H-1), 9.06 (d, 1H, J = 8.0 Hz, H-4) 9.79 (s, 1H, H-6); <13> C NMR (DMSO-d6) δ 89.4 (s), 115.4 (s), 115.7 (d), 116.9 (d), 119.8 (s), 120.3 (d ), 121.7 (s), 124.7 (d), 127.2 (s), 128.4 (d), 128.5 (d), 129.0 (d), 139.7 (d), 132.2 (s), 138.5 (s), 143.3 (d ), Anal. (C16H9N3) C, H, N.
ESEMPIO 7 EXAMPLE 7
11-ciano-3-metossi-isoindolo[2.1-a]chinossalina (3b) 11-cyano-3-methoxy-isoindole [2.1-a] quinoxaline (3b)
La preparazione dellintermedio di formula (4): 1-ciano-2-(2’-ammino-4’-metossi-fenil)-isoindolo è stata eseguita come nell'Esempio 3. L'intermedio (3 mmoli) è stato è stato disciolto in acido formico al 99% (10 ml) e tenuto a riflusso fino alla completa scomparsa del materiale di partenza, per un tempo totale di 10 h. La miscela di reazione è stata versata in acqua ghiacciata (150 mi) e il precipitato risultante è stato raccolto per filtrazione. Il solido ottenuto è stato purificato per flash-cromatografia utilizzando diclorometano a dare il prodotto del titolo, solido. The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-4'-methoxy-phenyl) -isoindole was carried out as in Example 3. The intermediate (3 mmol) was dissolved in 99% formic acid (10 ml) and refluxed until the complete disappearance of the starting material, for a total time of 10 h. The reaction mixture was poured into ice water (150ml) and the resulting precipitate was collected by filtration. The solid obtained was purified by flash chromatography using dichloromethane to give the title product, solid.
Resa 68%; p.f. 220-221°C; Yield 68%; m.p. 220-221 ° C;
IR: 2193 (CN) cm<-1>;<1>H NMR (DMSO-d6) δ 3.97 (s, 3H, CH3), 7.31 (dd, 1 H, J=9.6, 3.0 Hz, H-2), 7.46 (t, 1H, J=8.1 Hz, H-8), 7.53 (s, 1H, H-4), 7.58 (t, 1H, J=8.1 Hz, H-9), 7.90 (d, 1H, J= 8.1 Hz, H-10), 8.19 (d, 1H, J= 8.1 Hz, H-7), 9.06 (d, 1H, J= 9.6 Hz, H-1), 8.81 (s, 1H, H-6);<13>C NMR (DMSO-d6) δ 55.8 (q), 90.2 (s), 111.1 (d), 115.6 (s), 117.2 (d), 117.6 (d), 118.6 (d), 118.8 (d), 120.2 (s), 121.2 (s), 122.0 (s), 124.8 (d), 127.9 (d), 132.6 (s), 140.5 (s), 142.3 (d), 159.2 (s). Anal. (C17H1N3O) C, H, N. IR: 2193 (CN) cm <-1>; <1> H NMR (DMSO-d6) δ 3.97 (s, 3H, CH3), 7.31 (dd, 1 H, J = 9.6, 3.0 Hz, H-2) , 7.46 (t, 1H, J = 8.1 Hz, H-8), 7.53 (s, 1H, H-4), 7.58 (t, 1H, J = 8.1 Hz, H-9), 7.90 (d, 1H, J = 8.1 Hz, H-10), 8.19 (d, 1H, J = 8.1 Hz, H-7), 9.06 (d, 1H, J = 9.6 Hz, H-1), 8.81 (s, 1H, H- 6); <13> C NMR (DMSO-d6) δ 55.8 (q), 90.2 (s), 111.1 (d), 115.6 (s), 117.2 (d), 117.6 (d), 118.6 (d), 118.8 (d), 120.2 (s), 121.2 (s), 122.0 (s), 124.8 (d), 127.9 (d), 132.6 (s), 140.5 (s), 142.3 (d), 159.2 (s). Anal. (C17H1N3O) C, H, N.
ESEMPIO 8 EXAMPLE 8
11-ciano-2.3-dimetil-isoindolo[2.1-a]chinossalina (3c) 11-cyano-2.3-dimethyl-isoindole [2.1-a] quinoxaline (3c)
La preparazione dell'intermedio di formula (4): 1-ciano-2-(2’-ammino-2,3-dimetil-fenil)-isoindolo è stata eseguita come nell'Esempio 4. L’intermedio (3 mmoli) è stato è stato disciolto in acido formico al 99% (10 mi) e tenuto a riflusso fino alla completa scomparsa del materiale di partenza, cioè per un tempo totale di 30 h. La miscela di reazione è stata versata in acqua ghiacciata (150 mi) e il precipitato risultante è stato raccolto per filtrazione. Il solido ottenuto è stato purificato per flash-cromatografia utilizzando diclorometano a dare il prodotto del titolo, solido. The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-2,3-dimethyl-phenyl) -isoindole was carried out as in Example 4. The intermediate (3 mmol) is was dissolved in 99% formic acid (10 ml) and kept under reflux until the complete disappearance of the starting material, ie for a total time of 30 h. The reaction mixture was poured into ice water (150ml) and the resulting precipitate was collected by filtration. The solid obtained was purified by flash chromatography using dichloromethane to give the title product, solid.
Resa 71%; p.f. 220-221 °C; Yield 71%; m.p. 220-221 ° C;
IR: 2193 (CN) cm<-1>;<1>H NMR (DMSO-de) δ 2.45 (s, 3H, CH3), 2.54 (s, 3H, CH3), 7.49 (t, 1H, J=7.5 Hz, H-8), 7.60 (t, 1H, J=7.5 Hz, H-9), 7.85-7.88 (m, 2H, H-10 e H-4), 8.33 (d, 1H, J= 7.5 Hz, H-7), 8.85 (s, 1H, H-1), 9.44 (s, 1H, H-6);<13>C NMR (DMSO-d6) δ 19.2 (q), 20.1 (q), 111.2 (s), 113.1 (s), 115.2 (d), 116.6 (d), 119.0 (d), 119.4 (s), 121.0 (s), 124.0 (d), 124.9 (s), 127.4 (d), 130.0 (d), 132.0 (s), 136.7 (s), 137.3 (s), 138.2 (s), 141.0 (d). Anal. (C1eHi3N3) C, H, N. IR: 2193 (CN) cm <-1>; <1> H NMR (DMSO-de) δ 2.45 (s, 3H, CH3), 2.54 (s, 3H, CH3), 7.49 (t, 1H, J = 7.5 Hz, H-8), 7.60 (t, 1H, J = 7.5 Hz, H-9), 7.85-7.88 (m, 2H, H-10 and H-4), 8.33 (d, 1H, J = 7.5 Hz , H-7), 8.85 (s, 1H, H-1), 9.44 (s, 1H, H-6); <13> C NMR (DMSO-d6) δ 19.2 (q), 20.1 (q), 111.2 (s), 113.1 (s), 115.2 (d), 116.6 (d), 119.0 (d), 119.4 (s), 121.0 (s), 124.0 (d), 124.9 (s), 127.4 (d), 130.0 (d), 132.0 (s), 136.7 (s), 137.3 (s), 138.2 (s), 141.0 (d). Anal. (C1eHi3N3) C, H, N.
ESEMPIO 9 EXAMPLE 9
11-ciano-2.3-dicloro-isoindolo[2.1-a]chinossalina (3d) 11-cyano-2.3-dichloro-isoindole [2.1-a] quinoxaline (3d)
La preparazione dell’intermedio di formula (4): 1-ciano-2-(2’-ammino-2,3-dicloro-fenil)-isoindolo è stata eseguita come nell’Esempio 5. L'intermedio (3 mmoli) è stato è stato disciolto in acido formico al 99% (10 mi) e tenuto a riflusso fino alla completa scomparsa del materiale di partenza, cioè per un tempo totale di 15 h. La miscela di reazione è stata versata in acqua ghiacciata (150 mi) e il precipitato risultante è stato raccolto per filtrazione. Il solido ottenuto è stato purificato per flash-cromatografia utilizzando diclorometano a dare il prodotto del titolo, solido. The preparation of the intermediate of formula (4): 1-cyano-2- (2'-amino-2,3-dichloro-phenyl) -isoindole was carried out as in Example 5. The intermediate (3 mmol) is was dissolved in 99% formic acid (10 ml) and kept under reflux until the complete disappearance of the starting material, ie for a total time of 15 h. The reaction mixture was poured into ice water (150ml) and the resulting precipitate was collected by filtration. The solid obtained was purified by flash chromatography using dichloromethane to give the title product, solid.
Resa 75%; p.f. 279-281<º>C; Yield 75%; m.p. 279-281 <º> C;
IR: 2196 (CN) cm<'1>.<1>H NMR (CDCI3) δ 7.57 (t, 1H, J=8.3 Hz, H-8), 7.68 (t, 1H, J=8.3 Hz, H-9), 8.00 (d, 1H, J=8.3 Hz, H-10), 8.29 (d, 1H, J= 8.3 Hz, H-7), 8.31 (s, 1H, H-1), 9.35 (s, 1H, H-4), 9.46 (s, 1H, H-6);<13>C NMR (CDCI3) δ 86.9 (s), 96.5 (s), 112.5 (s), 117.6 (dx2), 119.1 (s), 121.7 (s), 125.8 (s), 128.8 (dx2), 129.7 (s), 131.8 (d), 143.3 (dx2), 145.1 (s), 152.3 (s). Anal. (C16H7CI2N3) C, H, N. IR: 2196 (CN) cm <'1>. <1> H NMR (CDCI3) δ 7.57 (t, 1H, J = 8.3 Hz, H-8), 7.68 (t, 1H, J = 8.3 Hz, H- 9), 8.00 (d, 1H, J = 8.3 Hz, H-10), 8.29 (d, 1H, J = 8.3 Hz, H-7), 8.31 (s, 1H, H-1), 9.35 (s, 1H, H-4), 9.46 (s, 1H, H-6); <13> C NMR (CDCI3) δ 86.9 (s), 96.5 (s), 112.5 (s), 117.6 (dx2), 119.1 (s ), 121.7 (s), 125.8 (s), 128.8 (dx2), 129.7 (s), 131.8 (d), 143.3 (dx2), 145.1 (s), 152.3 (s). Anal. (C16H7CI2N3) C, H, N.
Studio dell'attività antitumorale dei derivati isoindolo-chinossalinici Study of the antitumor activity of isoindole-quinoxaline derivatives
Saggi su linee cellulari - inibizione della crescita cellulare e ritotossicità Cell line assays - cell growth inhibition and retotoxicity
I derivati isoindolo-chinossalinici dell'invenzione sono stati valutato dal National Cancer Institute (NCI, Bethesda, USA), per mezzo del saggio antitumorale primario a singola dose su tre linee cellulari (MCF7-mammario, NCI-H460-polmonare a cellule non piccole, SF-268-SNC). In questo test, un composto è considerato attivo quando riduce la crescita di almeno una delle linee cellulare al 32% o meno (valori negativi indicano soppressione delle cellule). I risultati ottenuti sono riportati nella seguente Tabella 1. The isoindole-quinoxaline derivatives of the invention were evaluated by the National Cancer Institute (NCI, Bethesda, USA), by means of the single-dose primary antitumor assay on three cell lines (MCF7-mammary, NCI-H460-non-small cell lung , SF-268-SNC). In this test, a compound is considered active when it reduces the growth of at least one of the cell lines to 32% or less (negative values indicate cell suppression). The results obtained are reported in the following Table 1.
(segue tabella) TABELLA 1 (table follows) TABLE 1
Saggio antitumorale primario a singoia dose per le isoindolo-chinossaline Comp. Polmonare a cellule Single dose primary anticancer assay for isoindole-quinoxalines. Pulmonary Cell Comp.
Mammario SNC scelto Comp. non piccole NCI-MCF7 SF-268 per test H460 Mammary CNS selected Non-small Comp. NCI-MCF7 SF-268 for H460 test
60 cell. 60 cell.
2b 0 6 9 S 2c 2 27 55 s 2a 0 0 1 s 2e 0 10 13 s 2d 0 0 0 s 3a 91 107 94 N 3b 97 115 96 N 3c 78 97 83 N 3d 104 108 104 N 2b 0 6 9 S 2c 2 27 55 s 2a 0 0 1 s 2e 0 10 13 s 2d 0 0 0 s 3a 91 107 94 N 3b 97 115 96 N 3c 78 97 83 N 3d 104 108 104 N
I composti che hanno superato i criteri stabiliti dal NCI per l'attività in questo saggio sono stati automaticamente sottoposti allo screening contro l'intero panel di linee cellulari tumorali (Monks, A. et al. Feasibility of a High-Flux Anticancer Drug Screen Using a Diverse Panel of Cultured Human Tumor Celi Lines J. Natl. Cancer Inst., 1991, 83, 757-766). Tutti i derivati isoindolo-chinossalinici del tipo della formula (2) sono stati selezionati per la valutazione contro il panel completo di linee cellulari tumorali, che comprende circa 60 diverse linee cellulari neoplastiche umane, raggruppate in nove sottogruppi neoplastici: leucemia, tumore polmonare a cellule non piccole, tumore del colon; tumore del SNC, melanoma; carcinoma ovarico, tumore renale, carcinoma prostatico, carcinoma mammario. Compounds that passed NCI criteria for activity in this assay were automatically screened against the entire panel of tumor cell lines (Monks, A. et al. Feasibility of a High-Flux Anticancer Drug Screen Using a Diverse Panel of Cultured Human Tumor Cell Lines J. Natl. Cancer Inst., 1991, 83, 757-766). All isoindole-quinoxaline derivatives of the type of formula (2) were selected for evaluation against the complete panel of tumor cell lines, which includes about 60 different human neoplastic cell lines, grouped into nine neoplastic subgroups: leukemia, lung cell tumor not small, colon cancer; CNS tumor, melanoma; ovarian cancer, kidney cancer, prostate cancer, breast cancer.
I seguenti parametri sono stati determinati per ogni linea cellulare: Gl50(concentrazione che inibisce il 50% della crescita cellulare netta), TGI (inibizione della crescita totale) e LC50(concentrazione che causa il 50% di morte cellulare netta). Per ciascuno di questi parametri sono stati calcolati i valori medi del “mean graph midpoint" (MG_MID) (Boyd, M. R. e Paull, K. D., Some practical considerations and applications of thè National Cancer Institute in vitro anticancer drug discovery screen, Drug Dev. Res., 1995, 34, 91-109) The following parameters were determined for each cell line: Gl50 (concentration that inhibits 50% of net cell growth), TGI (inhibition of total growth) and LC50 (concentration that causes 50% of net cell death). For each of these parameters the mean values of the "mean graph midpoint" (MG_MID) were calculated (Boyd, M. R. and Paull, K. D., Some practical considerations and applications of the National Cancer Institute in vitro anticancer drug discovery screen, Drug Dev. Res ., 1995, 34, 91-109)
I dati ottenuti sono riportati nella Tabella 2 che segue. The data obtained are shown in Table 2 below.
(segue tabella) (table follows)
Una valutazione dei dati riportati in Tabella 2 ha evidenziato che i composti (2a-e) mostrano attività antineoplastica nei confronti di tutte le linee cellulari umane. Il composto più attivo è risultato essere il metossi-derivato (2a) (MG_MID = 7,32), seguito dal metil-derivato (2c), dal derivato non sostituito (2b) e dal derivato dimetiiico (2d), che hanno mostrato valori di MG_MID simili (5,28, 5,17 e 4,99). Il meno attivo era il dicloro-derivato (2e) (MG_MID = 4,74). An evaluation of the data reported in Table 2 showed that the compounds (2a-e) show antineoplastic activity against all human cell lines. The most active compound was found to be the methoxy-derivative (2a) (MG_MID = 7.32), followed by the methyl-derivative (2c), the unsubstituted derivative (2b) and the dimethyl derivative (2d), which showed values of similar MG_MIDs (5.28, 5.17 and 4.99). The least active was the dichloro-derivative (2e) (MG_MID = 4.74).
La Tabella 3 che segue riporta i valori di pGI50dei composti delle formule (2a-e) nei confronti di tutte le linee cellulari utilizzate nel test. Le isoindolo-chinossaline hanno mostrato un effetto inibitorio della crescita di tutte le linee cellulari tumorali a concentrazioni tra micromolari e nanomolari. Table 3 below reports the pGI50 values of the compounds of the formulas (2a-e) against all the cell lines used in the test. The isoindole-quinoxalines showed a growth inhibitory effect of all tumor cell lines at concentrations ranging from micromolar to nanomolar.
I derivati di formula (2), con l'eccezione di (2e), si sono mostrati particolarmente efficaci contro il sottogruppo della leucemia. In effetti, I valori di pGI50MG MID calcolati per il sottogruppo della leucemia sono sempre superiori alla media totale dei valori di MG_MID di tutte le linee cellulari (AMG_MID 0,24 - 0,51). The derivatives of formula (2), with the exception of (2e), have been shown to be particularly effective against the leukemia subgroup. Indeed, pGI50MG MID values calculated for the leukemia subgroup are always higher than the total mean of MG_MID values of all cell lines (AMG_MID 0.24 - 0.51).
II composto (2a) era selettivo verso l88% delle linee cellulari a valori di GI50dell'ordine di 10<-8>M. Le linee cellulari più sensibili sono HL-60(TB) (pGl507,94), SR (pGI508,00) e K562 (pGl508,05), che appartengono al sottogruppo della leucemia; MALME-3M (pGI508,00) e M14 (pGI508,00) del sottogruppo del melanoma e MDA-MB-435 (pGI508,35), che appartiene al sottogruppo del tumore mammario. Compound (2a) was selective towards 88% of cell lines at GI50 values of the order of 10 <-8> M. The most sensitive cell lines are HL-60 (TB) (pGl507.94), SR (pGI508.00) and K562 (pGl508.05), which belong to the leukemia subgroup; MALME-3M (pGI508.00) and M14 (pGI508.00) of the melanoma subgroup and MDA-MB-435 (pGI508.35), which belongs to the breast cancer subgroup.
Il composto (2c), recante un gruppo metilico, si è dimostrato selettivo rispetto al sottogruppo della leucemia, avendo tutte le linee cellulari del sottogruppo valori pGI50nel campo 5,70-5,92. Una risposta eccellente è stata anche osservata nel sottogruppo del tumore mammario. Le linee cellulari più sensibili sono risultate le MCF7 (pGIso >8,00), MDA-MB-231/ATCC (pGI505,62) e MDA-MB-435 (pGI505,50). Il derivato (2b) è risultato selettivamente attivo rispetto al carcinoma mammario e specialmente nei confronti di HS 578T (pGIso 6,28) e MDA-MB-231/ATCC (pGI506,02). Compound (2c), bearing a methyl group, proved to be selective with respect to the leukemia subgroup, having all cell lines of the subgroup pGI50 values in the range 5.70-5.92. Excellent response was also observed in the breast cancer subgroup. The most sensitive cell lines were MCF7 (pGIso> 8.00), MDA-MB-231 / ATCC (pGI505.62) and MDA-MB-435 (pGI505.50). Derivative (2b) was found to be selectively active with respect to breast cancer and especially against HS 578T (pGIso 6.28) and MDA-MB-231 / ATCC (pGI506.02).
Il dicloro-derivato (2e) ha mostrato un basso valore medio di pGI50(4,74), ma una buona selettività nei confronti della linea cellulare HCC-2998 (pGI506,09), appartenente al sottogruppo del tumore del colon. The dichloro-derivative (2e) showed a low mean value of pGI50 (4.74), but good selectivity towards the cell line HCC-2998 (pGI506.09), belonging to the colon cancer subgroup.
I dati ottenuti nella valutazione del TGI evidenziano che anche sotto questo profilo il composto più interessante è il (2a), sia in termini di pTGI (MG_MID = 5,87) che in termini di attività contro il maggior numero di linee cellulari (59), seguito, nell'ordine, da (2d), (2c) (2b) e (2e). A livello di LC50, la migliore risposta è stata osservata nel caso del composto (2a) nei confronti della linea cellulare HCC-2998 di tumore del colon (pLC506,30). The data obtained in the evaluation of the TGI show that also from this point of view the most interesting compound is (2a), both in terms of pTGI (MG_MID = 5.87) and in terms of activity against the greatest number of cell lines (59) , followed, in order, by (2d), (2c) (2b) and (2e). At the LC50 level, the best response was observed in the case of compound (2a) against the colon cancer cell line HCC-2998 (pLC506,30).
(segue tabella) (table follows)
<a>Dati ottenuti dall'“in vitro disease-oriented human tumor cells screen" del NCI. <a> Data obtained from the "in vitro disease-oriented human tumor cells screen" of the NCI.
<b>pGI50è il -log della concentrazione molare che inibisce il 50% della crescita cellulare netta.<c>Non determinato.<d>MG_MID = mean graph midpoint = valore medio aritmetico per tutte le linee cellulari tumorali saggiate. Se l’effetto indicato non era ottenibile entro l'intervallo di concentrazione usato, la concentrazione più alta saggiata è stata utilizzata per il calcolo. <b> pGI50 is the molar concentration -log that inhibits 50% of net cell growth. <c> Not determined. <d> MG_MID = mean graph midpoint = arithmetic mean value for all tumor cell lines tested. If the indicated effect was not obtainable within the concentration range used, the highest concentration tested was used for the calculation.
Effetto sulla polimerizzazione dei microtubuli in vitro Effect on the polymerization of microtubules in vitro
I test di immunofluorescenza e i saggi di determinazione dell'indice mitotico condotti sulle isoindolo-chinossaline della presente invenzione, e in modo particolare sui composti di formula (2a), hanno suggerito che questi composti interferiscono con la formazione o con la stabilità del reticolo di microtubuli nelle cellule, ed è noto che i microtubuli sono i maggiori responsabili della divisione mitotica. Immunofluorescence tests and mitotic index assays conducted on the isoindole-quinoxalines of the present invention, and in particular on the compounds of formula (2a), have suggested that these compounds interfere with the formation or stability of the microtubule lattice. in cells, and microtubules are known to be most responsible for mitotic division.
Pertanto, è stata condotta una ulteriore sperimentazione per valutare gli effetti dei composti dell'invenzione sulla polimerizzazione della proteina dei microtubuli isolata da cervello porcino. Il saggio utilizza un composto fluorescente che si lega alla tubulina e ai microtubuli: la polimerizzazione è seguita da incremento della fluorescenza a causa dell’incorporazione di un reporter fluorescente nei microtubuli quando la polimerizzazione ha luogo (Bonne D., et al. 4',6-diamidino-2-phenylindole, a fluorescent probe for tubulin and microtubule. J. Biol. Chem. Therefore, a further experimentation was carried out to evaluate the effects of the compounds of the invention on the polymerization of the microtubule protein isolated from porcine brain. The assay uses a fluorescent compound that binds to tubulin and microtubules: polymerization is followed by increased fluorescence due to the incorporation of a fluorescent reporter into the microtubules when polymerization takes place (Bonne D., et al. 4 ', 6-diamidino-2-phenylindole, a fluorescent probe for tubulin and microtubule. J. Biol. Chem.
1985, 260: 2819-2825). 1985, 260: 2819-2825).
Per la sperimentazione è stato utilizzato un kit per il saggio della polimerizzazione dei microtubuli (Cytoskeleton USA) seguendo il protocollo raccomandato dal produttore. Il saggio è stato eseguito in piastre da microtitolazione da 96 pozzetti e la reazione è stata iniziata attraverso l'aggiunta di tubulina. La piastra è stata incubata a 37°C in un lettore di micropiastre a fluorescenza (Fluoroskan Ascent FL Labsystems) e la misura di fluorescenza (λex 355 nm; λm450 nm) è stata determinata ogni minuto per 60 min. Come composti di riferimento, sono stati usati taxol (Cytoskeleton USA) e colchicina (Sigma). For the experimentation, a kit for the microtubule polymerization assay (Cytoskeleton USA) was used following the protocol recommended by the manufacturer. The assay was performed in 96-well microtiter plates and the reaction was initiated through the addition of tubulin. The plate was incubated at 37 ° C in a fluorescence microplate reader (Fluoroskan Ascent FL Labsystems) and the fluorescence measurement (λex 355 nm; λm450 nm) was determined every minute for 60 min. As reference compounds, taxol (Cytoskeleton USA) and colchicine (Sigma) were used.
I risultati, che sono mostrati nella Figura 1, evidenziano che nel campione di controllo (tubulina senza l'aggiunta di composti in prova) la fluorescenza aumenta nel tempo. Al contrario, tutti e quattro i campioni studiati inducono chiaramente un'inibizione, dipendente dalla concentrazione, della polimerizzazione di tubulina. The results, which are shown in Figure 1, show that in the control sample (tubulin without the addition of test compounds) the fluorescence increases over time. In contrast, all four samples studied clearly induce a concentration-dependent inhibition of tubulin polymerization.
Una completa inibizione dell’aggregazione di microtubuli è stata osservata per il composto di formula (2a) alia concentrazione 5 μm. A complete inhibition of microtubule aggregation was observed for the compound of formula (2a) at a concentration of 5 μm.
In successive prove è stato investigato l’effetto del composto (2a) su microtubuli preassemblati. Nessuno dei composti saggiati è risultato in grado di indurre la depolimerizzazione dei microtubuli (dati non mostrati), indicando che l'azione di questi composti è strettamente correlata all'interferenza con l'aggregazione dei microtubuli. In subsequent tests, the effect of the compound (2a) on pre-assembled microtubules was investigated. None of the compounds tested was found to induce microtubule depolymerization (data not shown), indicating that the action of these compounds is closely related to interference with microtubule aggregation.
Saggio della topoisomerasi Topoisomerase assay
Esperimenti di rilassamento del DNA sono stati effettuati sia con topoisomerasi I che con topoisomerasi II. DNA relaxation experiments were performed with both topoisomerase I and topoisomerase II.
DNA supercoiled di pBR322 (Fermentas) è stato incubato con 5 unità di topoisomerasi I umana a 37°C per 30 min in tampone di rilassamento (50 mM Tris-HCI pH 7,8, 50 mM KCI, 10 mM MgCl2, 1 mM ditiotreitolo, 1 mM EDTA) in presenza di concentrazioni variabili (da 0,5 a 50 μΜ) del composto in studio. PBR322 (Fermentas) supercoiled DNA was incubated with 5 units of human topoisomerase I at 37 ° C for 30 min in relaxation buffer (50 mM Tris-HCI pH 7.8, 50 mM KCI, 10 mM MgCl2, 1 mM dithiothreitol , 1 mM EDTA) in the presence of varying concentrations (from 0.5 to 50 μΜ) of the compound under study.
Le reazioni sono state terminate per aggiunta di SDS allo 0,25% e di Proteinasi K a 250 pg/ml. Campioni di DNA sono stati quindi aggiunti al tampone di carica e sottoposti ad elettroforesi in un gel di agarosio all’1% a temperatura ambiente durante la notte a 20 V. I gel sono stati colorati con bromuro di etidio (1 mg/ml), lavati e dopo la colo razione il gel è stato lavato con acqua e le bande di DNA sono state determinate sotto radiazione UV con un transilluminatore. Sono state fatte fotografie con una macchina digitale Kodak DC256 e la quantificazione delle bande è stata ottenuta per mezzo del software analizzatore di immagine Quantity One (BIO RAD, Milano Italia). Simili esperimenti sono stati condotti utilizzando gel di agarosio contenenti etidio. The reactions were terminated by addition of 0.25% SDS and of Proteinase K at 250 pg / ml. DNA samples were then added to the charge buffer and electrophoresed in a 1% agarose gel at room temperature overnight at 20 V. The gels were stained with ethidium bromide (1 mg / mL), washed and after coloring the gel was washed with water and the DNA bands were determined under UV radiation with a transilluminator. Photographs were taken with a Kodak DC256 digital camera and the quantification of the bands was obtained by means of the Quantity One image analyzer software (BIO RAD, Milan Italy). Similar experiments were conducted using ethidium-containing agarose gels.
Il gel mostrato nella Figura 2 (sezione A) indica che a basse concentrazioni l’intensità della banda che migra più lentamente (corrispondente a DNA nicked totalmente rilassato) è aumentata significativamente, mentre a concentrazione maggiore il rilassamento del DNA è stato inibito, come osservato con l'avvelenante di riferimento per la toposomerasi I, la camptotecina, usata alla concentrazione di 20 μΜ. The gel shown in Figure 2 (section A) indicates that at low concentrations the intensity of the slower migrating band (corresponding to totally relaxed nicked DNA) increased significantly, while at higher concentration DNA relaxation was inhibited, as observed. with the reference poisoner for toposomerase I, camptothecin, used at a concentration of 20 μΜ.
Per escludere un effetto aspecifico dovuto all'interazione diretta del composto (2a) con il DNA, il test di rilassamento è stato ripetuto su bromuro di etidio contenente gel di agarosio, come mostrato nella Figura 2 (sezione B). In queste condizioni è stata osservata la presenza di molecole di DNA nicked, indicando che il composto (2a) può stimolare la scissione del DNA da parte della topoisomerasi I. To exclude a non-specific effect due to the direct interaction of the compound (2a) with the DNA, the relaxation test was repeated on ethidium bromide containing agarose gel, as shown in Figure 2 (section B). The presence of nicked DNA molecules was observed under these conditions, indicating that compound (2a) can stimulate DNA cleavage by topoisomerase I.
Analoghi esperimenti sono stati condotti con la topoisomerasi II umana, e i risultati hanno indicato che il composto (2a) non ha nessun effetto su questo enzima (dati non mostrati). Similar experiments were conducted with human topoisomerase II, and the results indicated that compound (2a) has no effect on this enzyme (data not shown).
Conclusioni Conclusions
I risultati complessivi della sperimentazione parzialmente illustrata in quanto precede mostrano che i derivati isoindolo[2,1-a]chinossalinici dell’invenzione possiedono un'elevata attività citotossica, e che essi agiscono attraverso l’inibizione della polimerizzazione della tubulina e dell’attività della topoisomerasi I. The overall results of the experiment partially illustrated above show that the isoindole [2,1-a] quinoxaline derivatives of the invention possess a high cytotoxic activity, and that they act through the inhibition of tubulin polymerization and the activity of topoisomerase I.
I composti dell'invenzione si sono mostrati in grado di inibire la polimerizzazione tubulinica in vitro in maniera dipendente dalla concentrazione, in modo simile alla colchicina e alla vinblastina, con ciò indicando che tali composti possono essere classificati come agenti depolimerizzanti dei microtubuli. Inoltre, la determinazione dell'indice mitotico e gli studi di immunofluorescenza confermano l’interazione con la tubulina. È da notare, a questo proposito, che la struttura dei derivati isoindolo-chinossalinici dell’invenzione non assomiglia a nessuna delle strutture di noti agenti antimitotici. The compounds of the invention have been shown to inhibit tubulin polymerization in vitro in a concentration dependent manner, similar to colchicine and vinblastine, thereby indicating that such compounds can be classified as microtubule depolymerizing agents. Furthermore, the determination of the mitotic index and immunofluorescence studies confirm the interaction with tubulin. It should be noted, in this regard, that the structure of the isoindole-quinoxaline derivatives of the invention does not resemble any of the structures of known antimitotic agents.
I risultati spettrofotometrici indicano che il composto di formula (2a) si lega al DNA per intercalazione, e conseguentemente la macromolecola può essere considerata un potenziale target di questa classe di composti. In letteratura è noto che gli intercalatori di DNA possono essere inibitori della topoisomerasi I e/o della topoisomerasi II. Utilizzando un saggio di rilassamento del DNA è stato dimostrato che il composto preferito della presente invenzione inibisce l’attività catalitica della topoisomerasi I ma non quella della topoisomerasi II. The spectrophotometric results indicate that the compound of formula (2a) binds to DNA by intercalation, and consequently the macromolecule can be considered a potential target of this class of compounds. In the literature it is known that DNA intercalators can be inhibitors of topoisomerase I and / or topoisomerase II. Using a DNA relaxation assay it has been shown that the preferred compound of the present invention inhibits the catalytic activity of topoisomerase I but not that of topoisomerase II.
In conclusione, i risultati sopra sintetizzati dimostrano l’attività antitumorale in vitro dei derivati isoindolo[2,1-a]chinossalinici dell'invenzione, e in modo particolare quella del derivato isoindolo-chinossalinico di formula (2a), attività che appare esplicarsi attraverso due distinti meccanismi d’azione. In conclusion, the results summarized above demonstrate the in vitro antitumor activity of the isoindole [2,1-a] quinoxaline derivatives of the invention, and in particular that of the isoindole-quinoxaline derivative of formula (2a), an activity which appears to be expressed through two distinct mechanisms of action.
La presente invenzione è stata descritta con riferimento ad alcune sue forme di realizzazione specifiche, ma è da intendersi che variazioni o modifiche potranno essere ad essa apportate dagli esperti nel ramo senza per questo uscire dal relativo ambito di protezione. The present invention has been described with reference to some of its specific embodiments, but it is to be understood that variations or modifications may be made to it by those skilled in the art without thereby departing from the relative scope of protection.
Claims (21)
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT000518A ITRM20060518A1 (en) | 2006-10-02 | 2006-10-02 | ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE |
| PCT/IT2007/000689 WO2008041264A1 (en) | 2006-10-02 | 2007-10-02 | Isoindolo-quinoxaline derivatives having antitumor activity, process for their production and their use |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| IT000518A ITRM20060518A1 (en) | 2006-10-02 | 2006-10-02 | ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| ITRM20060518A1 true ITRM20060518A1 (en) | 2008-04-03 |
Family
ID=39113966
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| IT000518A ITRM20060518A1 (en) | 2006-10-02 | 2006-10-02 | ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE |
Country Status (2)
| Country | Link |
|---|---|
| IT (1) | ITRM20060518A1 (en) |
| WO (1) | WO2008041264A1 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101747207B1 (en) * | 2013-05-29 | 2017-06-15 | 희성소재 (주) | Aromatic fused ring compound and organic light emitting device using the same |
| ITRM20130588A1 (en) * | 2013-10-25 | 2015-04-26 | Univ Palermo | DERIVATIVES OF CHINOSALALIN, METHODS FOR THEIR PRODUCTION AND THEIR USE AS ANTI-HUMAN AGENTS. |
| KR20230034222A (en) | 2020-06-11 | 2023-03-09 | 씨에이치디아이 파운데이션, 인코포레이티드 | Heterocyclic compounds and imaging agents for imaging huntingtin protein |
| CN115010715B (en) * | 2022-06-24 | 2023-07-21 | 河南师范大学 | Synthesis method and anticancer activity of 3- (indole-2-yl) succinimide and indolobenzodiazepine compounds |
| CN115819424A (en) * | 2022-12-05 | 2023-03-21 | 重庆文理学院 | Indazolo-quinoxaline derivative, and synthetic method and application thereof |
-
2006
- 2006-10-02 IT IT000518A patent/ITRM20060518A1/en unknown
-
2007
- 2007-10-02 WO PCT/IT2007/000689 patent/WO2008041264A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2008041264A1 (en) | 2008-04-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR102075886B1 (en) | Novel pyrazolo [3,4-d] pyrimidine compounds or salts thereof | |
| JP6919922B2 (en) | FGFR4 inhibitor, its manufacturing method and pharmaceutical application | |
| Ohashi et al. | Discovery of pyrrolo [3, 2-c] quinoline-4-one derivatives as novel hedgehog signaling inhibitors | |
| EP2310014B1 (en) | Tricyclic compounds having antimitotic and/or antitumor activity and methods of use thereof | |
| AU778735B2 (en) | Fused azepinone cyclin dependent kinase inhibitors | |
| Al-Issa | Synthesis and anticancer activity of some fused pyrimidines and related heterocycles | |
| Wang et al. | Synthesis and in vitro antiproliferative activity of new 11-aminoalkylamino-substituted 5H-and 6H-indolo [2, 3-b] quinolines; structure–activity relationships of neocryptolepines and 6-methyl congeners | |
| CA2647036C (en) | Tetrahydropyridothienopyrimidine compounds and methods of use thereof | |
| Abdel-hafez | Benzimidazole condensed ring systems: New synthesis and antineoplastic activity of substituted 3, 4-dihydro-and 1, 2, 3, 4-tetrahydro-benzo [4, 5] imidazo [1, 2-a] pyrinnidine derivatives | |
| WO2009077956A2 (en) | HETEROCYCLIC INHIBITORS OF AN Hh-SIGNAL CASCADE, MEDICINAL COMPOSITIONS BASED THEREON AND METHODS FOR TREATING DISEASES CAUSED BY THE ABERRANT ACTIVITY OF AN Hh-SIGNAL SYSTEM | |
| JP2002525285A (en) | Pyrrolobenzodiazepines | |
| ES2529046T3 (en) | Preparation procedures for quinolone analogues | |
| Rostom et al. | Synthesis and biological evaluation of some polymethoxylated fused pyridine ring systems as antitumor agents | |
| Xin et al. | Design, synthesis, and evaluation of pyrrolo [2, 1-f][1, 2, 4] triazine derivatives as novel hedgehog signaling pathway inhibitors | |
| JP2008509971A (en) | Benzothienopyridine for use as an inhibitor of EG5 kinesin | |
| ITRM20060518A1 (en) | ISOINDOLO-KINOSALALINE DERIVATIVES WITH ANTITUMORAL ACTIVITY PROCEDURE FOR THEIR PRODUCTION AND THEIR USE | |
| JP2006517949A (en) | Tetracyclic pyrazole derivatives as kinase inhibitors, methods for producing the derivatives, and pharmaceutical compositions containing the derivatives | |
| Stefańska et al. | 2, 7-Dihydro-3H-pyridazino [5, 4, 3-kl] acridin-3-one derivatives, novel type of cytotoxic agents active on multidrug-resistant cell lines. Synthesis and biological evaluation | |
| Shaaban et al. | Synthesis, characterisation and biological screening of some 2-substituted benzimidazole derivatives as potential anticancer agents | |
| Bu et al. | Synthesis and cytotoxic activity of N-[(alkylamino) alkyl] carboxamide derivatives of 7-oxo-7H-benz [de] anthracene, 7-oxo-7H-naphtho [1, 2, 3-de] quinoline, and 7-oxo-7H-benzo [e] perimidine | |
| Raju et al. | Synthesis, characterization and in vitro antiproliferative effects of novel 5-amino pyrazole derivatives against breast cancer cell lines | |
| Hanna et al. | Synthesis and antitumor evaluation of some novel pyrrolizine derivatives | |
| CN102229563A (en) | 4-amino quinoline derivative, preparation method and application thereof | |
| Al‐Taifi et al. | Synthesis and Characterization of New Heterocyclic Compounds Containing Thienylbenzo [h] Quinoline Moiety | |
| El-Gazzar et al. | Anticancer evaluation and docking study of new bifunctional phthalazine derivatives |