CA3091950A1 - 7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer - Google Patents
7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer Download PDFInfo
- Publication number
- CA3091950A1 CA3091950A1 CA3091950A CA3091950A CA3091950A1 CA 3091950 A1 CA3091950 A1 CA 3091950A1 CA 3091950 A CA3091950 A CA 3091950A CA 3091950 A CA3091950 A CA 3091950A CA 3091950 A1 CA3091950 A1 CA 3091950A1
- Authority
- CA
- Canada
- Prior art keywords
- amino
- methyl
- oxo
- purine
- compound
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000011282 treatment Methods 0.000 title claims abstract description 133
- 201000007270 liver cancer Diseases 0.000 title claims abstract description 96
- 208000014018 liver neoplasm Diseases 0.000 title claims abstract description 96
- 238000011321 prophylaxis Methods 0.000 title claims abstract description 57
- -1 7-substituted sulfonimidoylpurinone compounds Chemical class 0.000 title claims description 179
- 150000001875 compounds Chemical class 0.000 claims abstract description 844
- 150000003839 salts Chemical class 0.000 claims abstract description 54
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 claims description 120
- 125000003854 p-chlorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C([H])=C1Cl 0.000 claims description 110
- 238000000034 method Methods 0.000 claims description 101
- 238000002360 preparation method Methods 0.000 claims description 101
- WNOQDUGFAXKMOI-UHFFFAOYSA-N purine-7-carboxamide Chemical compound C1=NC=C2N(C(=O)N)C=NC2=N1 WNOQDUGFAXKMOI-UHFFFAOYSA-N 0.000 claims description 81
- 125000004800 4-bromophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C([H])=C1Br 0.000 claims description 78
- 239000000203 mixture Substances 0.000 claims description 69
- 230000003042 antagnostic effect Effects 0.000 claims description 67
- 239000003814 drug Substances 0.000 claims description 57
- 101000611936 Homo sapiens Programmed cell death protein 1 Proteins 0.000 claims description 50
- 125000000999 tert-butyl group Chemical group [H]C([H])([H])C(*)(C([H])([H])[H])C([H])([H])[H] 0.000 claims description 47
- MLDQJTXFUGDVEO-UHFFFAOYSA-N BAY-43-9006 Chemical compound C1=NC(C(=O)NC)=CC(OC=2C=CC(NC(=O)NC=3C=C(C(Cl)=CC=3)C(F)(F)F)=CC=2)=C1 MLDQJTXFUGDVEO-UHFFFAOYSA-N 0.000 claims description 46
- 239000005511 L01XE05 - Sorafenib Substances 0.000 claims description 46
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 46
- 229960003787 sorafenib Drugs 0.000 claims description 44
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 claims description 42
- 102100040678 Programmed cell death protein 1 Human genes 0.000 claims description 41
- 125000001255 4-fluorophenyl group Chemical group [H]C1=C([H])C(*)=C([H])C([H])=C1F 0.000 claims description 40
- 206010073071 hepatocellular carcinoma Diseases 0.000 claims description 37
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 claims description 35
- 239000004037 angiogenesis inhibitor Substances 0.000 claims description 34
- XBDQKXXYIPTUBI-UHFFFAOYSA-N Propionic acid Chemical compound CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 31
- 238000002648 combination therapy Methods 0.000 claims description 31
- 125000001449 isopropyl group Chemical group [H]C([H])([H])C([H])(*)C([H])([H])[H] 0.000 claims description 29
- 231100000844 hepatocellular carcinoma Toxicity 0.000 claims description 23
- 239000008194 pharmaceutical composition Substances 0.000 claims description 22
- 229910052736 halogen Inorganic materials 0.000 claims description 19
- BOUNFBOFBGBYBT-UHFFFAOYSA-N purin-8-one Chemical compound C1=NC=NC2=NC(=O)N=C21 BOUNFBOFBGBYBT-UHFFFAOYSA-N 0.000 claims description 18
- 229960000397 bevacizumab Drugs 0.000 claims description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 16
- 125000004803 chlorobenzyl group Chemical group 0.000 claims description 16
- 125000004175 fluorobenzyl group Chemical group 0.000 claims description 16
- 229960003852 atezolizumab Drugs 0.000 claims description 15
- HBAQYPYDRFILMT-UHFFFAOYSA-N 8-[3-(1-cyclopropylpyrazol-4-yl)-1H-pyrazolo[4,3-d]pyrimidin-5-yl]-3-methyl-3,8-diazabicyclo[3.2.1]octan-2-one Chemical class C1(CC1)N1N=CC(=C1)C1=NNC2=C1N=C(N=C2)N1C2C(N(CC1CC2)C)=O HBAQYPYDRFILMT-UHFFFAOYSA-N 0.000 claims description 12
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims description 12
- 125000006278 bromobenzyl group Chemical group 0.000 claims description 12
- 125000000623 heterocyclic group Chemical group 0.000 claims description 12
- 125000006178 methyl benzyl group Chemical group 0.000 claims description 12
- 125000001436 propyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 12
- 125000002393 azetidinyl group Chemical group 0.000 claims description 11
- 125000000719 pyrrolidinyl group Chemical group 0.000 claims description 11
- 108010074708 B7-H1 Antigen Proteins 0.000 claims description 10
- 229950002916 avelumab Drugs 0.000 claims description 10
- 208000006990 cholangiocarcinoma Diseases 0.000 claims description 10
- 229950009791 durvalumab Drugs 0.000 claims description 10
- 125000005843 halogen group Chemical group 0.000 claims description 10
- 150000002367 halogens Chemical class 0.000 claims description 10
- 229960003301 nivolumab Drugs 0.000 claims description 10
- 125000001424 substituent group Chemical group 0.000 claims description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 9
- 239000002147 L01XE04 - Sunitinib Substances 0.000 claims description 9
- 239000002138 L01XE21 - Regorafenib Substances 0.000 claims description 9
- 229960004836 regorafenib Drugs 0.000 claims description 9
- FNHKPVJBJVTLMP-UHFFFAOYSA-N regorafenib Chemical compound C1=NC(C(=O)NC)=CC(OC=2C=C(F)C(NC(=O)NC=3C=C(C(Cl)=CC=3)C(F)(F)F)=CC=2)=C1 FNHKPVJBJVTLMP-UHFFFAOYSA-N 0.000 claims description 9
- WINHZLLDWRZWRT-ATVHPVEESA-N sunitinib Chemical compound CCN(CC)CCNC(=O)C1=C(C)NC(\C=C/2C3=CC(F)=CC=C3NC\2=O)=C1C WINHZLLDWRZWRT-ATVHPVEESA-N 0.000 claims description 9
- 229960001796 sunitinib Drugs 0.000 claims description 9
- JZWSKEUHEWPEOH-UHFFFAOYSA-N 6-amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound NC1=C2N(C(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)C(=O)N(CCOC)CCOC JZWSKEUHEWPEOH-UHFFFAOYSA-N 0.000 claims description 8
- VAJCYQHLYBTSHG-UHFFFAOYSA-N ethyl n,n-diethylcarbamate Chemical compound CCOC(=O)N(CC)CC VAJCYQHLYBTSHG-UHFFFAOYSA-N 0.000 claims description 8
- PQKXVPODMYXMCP-UHFFFAOYSA-N ethyl n-methyl-n-propylcarbamate Chemical compound CCCN(C)C(=O)OCC PQKXVPODMYXMCP-UHFFFAOYSA-N 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 8
- ONIBWKKTOPOVIA-UHFFFAOYSA-M prolinate Chemical compound [O-]C(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-M 0.000 claims description 8
- OYNPDXXPHPYBJE-UHFFFAOYSA-N 6-amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound NC1=C2N(C(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)C(=O)N(CCOC)CC OYNPDXXPHPYBJE-UHFFFAOYSA-N 0.000 claims description 7
- 125000003386 piperidinyl group Chemical group 0.000 claims description 7
- 208000006359 hepatoblastoma Diseases 0.000 claims description 6
- 201000009030 Carcinoma Diseases 0.000 claims description 5
- 206010067388 Hepatic angiosarcoma Diseases 0.000 claims description 5
- 230000002440 hepatic effect Effects 0.000 claims description 5
- 125000000959 isobutyl group Chemical group [H]C([H])([H])C([H])(C([H])([H])[H])C([H])([H])* 0.000 claims description 5
- 201000010995 liver angiosarcoma Diseases 0.000 claims description 5
- 230000001394 metastastic effect Effects 0.000 claims description 5
- 206010061289 metastatic neoplasm Diseases 0.000 claims description 5
- 125000000484 butyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 claims description 4
- 125000004193 piperazinyl group Chemical group 0.000 claims description 4
- 102000008096 B7-H1 Antigen Human genes 0.000 claims 6
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 claims 1
- GCEQCCDAWABHRO-UHFFFAOYSA-N 2-[[6-amino-9-benzyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methylamino]ethyl N-butyl-N-methylcarbamate Chemical compound C(CCC)N(C(OCCN(C)C(=O)N1C(N(C2=NC(=NC(=C12)N)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)=O)C GCEQCCDAWABHRO-UHFFFAOYSA-N 0.000 claims 1
- 239000000651 prodrug Substances 0.000 abstract description 67
- 229940002612 prodrug Drugs 0.000 abstract description 67
- 239000000543 intermediate Substances 0.000 description 207
- IAZDPXIOMUYVGZ-WFGJKAKNSA-N Dimethyl sulfoxide Chemical compound [2H]C([2H])([2H])S(=O)C([2H])([2H])[2H] IAZDPXIOMUYVGZ-WFGJKAKNSA-N 0.000 description 172
- 238000001819 mass spectrum Methods 0.000 description 124
- 239000007787 solid Substances 0.000 description 95
- 238000005160 1H NMR spectroscopy Methods 0.000 description 78
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 64
- 206010028980 Neoplasm Diseases 0.000 description 62
- 239000000243 solution Substances 0.000 description 61
- 210000004027 cell Anatomy 0.000 description 52
- YEMRRHHMLASGRQ-UHFFFAOYSA-N n-methyl-n-propylcarbamoyl chloride Chemical compound CCCN(C)C(Cl)=O YEMRRHHMLASGRQ-UHFFFAOYSA-N 0.000 description 46
- 238000000746 purification Methods 0.000 description 44
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 36
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 34
- 239000011541 reaction mixture Substances 0.000 description 30
- 238000006243 chemical reaction Methods 0.000 description 29
- RFDSJHHLGFFVHD-UHFFFAOYSA-N tert-butyl n-(2-hydroxyethyl)-n-methylcarbamate Chemical compound OCCN(C)C(=O)OC(C)(C)C RFDSJHHLGFFVHD-UHFFFAOYSA-N 0.000 description 29
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 24
- 239000003921 oil Substances 0.000 description 24
- 239000012267 brine Substances 0.000 description 23
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 23
- 241000699670 Mus sp. Species 0.000 description 22
- 239000012044 organic layer Substances 0.000 description 22
- 210000001744 T-lymphocyte Anatomy 0.000 description 21
- 210000004369 blood Anatomy 0.000 description 20
- 239000008280 blood Substances 0.000 description 20
- 210000004185 liver Anatomy 0.000 description 20
- 210000004881 tumor cell Anatomy 0.000 description 20
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 19
- 235000011089 carbon dioxide Nutrition 0.000 description 19
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 18
- NIDZUMSLERGAON-UHFFFAOYSA-N ethyl 2-(methylamino)acetate;hydron;chloride Chemical compound Cl.CCOC(=O)CNC NIDZUMSLERGAON-UHFFFAOYSA-N 0.000 description 18
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 18
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 17
- 239000000843 powder Substances 0.000 description 17
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 16
- 239000007832 Na2SO4 Substances 0.000 description 16
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 16
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 16
- 239000012230 colorless oil Substances 0.000 description 16
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 16
- 229910052938 sodium sulfate Inorganic materials 0.000 description 16
- 235000011152 sodium sulphate Nutrition 0.000 description 16
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 15
- 229930182816 L-glutamine Natural products 0.000 description 15
- GVWISOJSERXQBM-UHFFFAOYSA-N n-methylpropan-1-amine Chemical compound CCCNC GVWISOJSERXQBM-UHFFFAOYSA-N 0.000 description 15
- 239000006228 supernatant Substances 0.000 description 15
- 102100024213 Programmed cell death 1 ligand 2 Human genes 0.000 description 14
- 238000000926 separation method Methods 0.000 description 14
- 101100407308 Mus musculus Pdcd1lg2 gene Proteins 0.000 description 13
- 108700030875 Programmed Cell Death 1 Ligand 2 Proteins 0.000 description 13
- 238000004296 chiral HPLC Methods 0.000 description 13
- HIHOEGPXVVKJPP-JTQLQIEISA-N 5-fluoro-2-[[(1s)-1-(5-fluoropyridin-2-yl)ethyl]amino]-6-[(5-methyl-1h-pyrazol-3-yl)amino]pyridine-3-carbonitrile Chemical compound N([C@@H](C)C=1N=CC(F)=CC=1)C(C(=CC=1F)C#N)=NC=1NC=1C=C(C)NN=1 HIHOEGPXVVKJPP-JTQLQIEISA-N 0.000 description 12
- BMHHQPYFORGXQA-UHFFFAOYSA-N 7h-purine-2-carboxamide Chemical compound NC(=O)C1=NC=C2N=CNC2=N1 BMHHQPYFORGXQA-UHFFFAOYSA-N 0.000 description 12
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 12
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 12
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 12
- 238000005481 NMR spectroscopy Methods 0.000 description 12
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- 150000001413 amino acids Chemical group 0.000 description 12
- 201000011510 cancer Diseases 0.000 description 12
- 238000001727 in vivo Methods 0.000 description 12
- 230000035755 proliferation Effects 0.000 description 12
- 238000005070 sampling Methods 0.000 description 12
- 235000017557 sodium bicarbonate Nutrition 0.000 description 12
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 12
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 description 12
- 230000004083 survival effect Effects 0.000 description 12
- PHDIJLFSKNMCMI-ITGJKDDRSA-N (3R,4S,5R,6R)-6-(hydroxymethyl)-4-(8-quinolin-6-yloxyoctoxy)oxane-2,3,5-triol Chemical compound OC[C@@H]1[C@H]([C@@H]([C@H](C(O1)O)O)OCCCCCCCCOC=1C=C2C=CC=NC2=CC=1)O PHDIJLFSKNMCMI-ITGJKDDRSA-N 0.000 description 11
- 101000652482 Homo sapiens TBC1 domain family member 8 Proteins 0.000 description 11
- 102100030302 TBC1 domain family member 8 Human genes 0.000 description 11
- 230000000694 effects Effects 0.000 description 11
- LCRTWYJHQIYYMR-UHFFFAOYSA-N ethyl n-butyl-n-methylcarbamate Chemical compound CCCCN(C)C(=O)OCC LCRTWYJHQIYYMR-UHFFFAOYSA-N 0.000 description 11
- 239000003981 vehicle Substances 0.000 description 11
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 10
- 239000003795 chemical substances by application Substances 0.000 description 10
- 238000011260 co-administration Methods 0.000 description 10
- WGTHNNRORWFTFA-UHFFFAOYSA-N ethyl pyrrolidine-1-carboxylate Chemical compound CCOC(=O)N1CCCC1 WGTHNNRORWFTFA-UHFFFAOYSA-N 0.000 description 10
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 10
- 210000003240 portal vein Anatomy 0.000 description 10
- 230000011664 signaling Effects 0.000 description 10
- 101000669402 Homo sapiens Toll-like receptor 7 Proteins 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 9
- 241000700159 Rattus Species 0.000 description 9
- 239000002253 acid Substances 0.000 description 9
- 208000035475 disorder Diseases 0.000 description 9
- 102000048362 human PDCD1 Human genes 0.000 description 9
- 229920006395 saturated elastomer Polymers 0.000 description 9
- 102000003945 NF-kappa B Human genes 0.000 description 8
- 108010057466 NF-kappa B Proteins 0.000 description 8
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 8
- 239000000427 antigen Substances 0.000 description 8
- 108091007433 antigens Proteins 0.000 description 8
- 102000036639 antigens Human genes 0.000 description 8
- 238000011284 combination treatment Methods 0.000 description 8
- 201000010099 disease Diseases 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 8
- 210000001853 liver microsome Anatomy 0.000 description 8
- 238000010172 mouse model Methods 0.000 description 8
- XZVYDRLPXWFRIS-UHFFFAOYSA-N n-ethyl-n-methylcarbamoyl chloride Chemical compound CCN(C)C(Cl)=O XZVYDRLPXWFRIS-UHFFFAOYSA-N 0.000 description 8
- 229960002621 pembrolizumab Drugs 0.000 description 8
- 239000000047 product Substances 0.000 description 8
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 description 8
- 102000005962 receptors Human genes 0.000 description 8
- 108020003175 receptors Proteins 0.000 description 8
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 7
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 7
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 7
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 7
- 102000002689 Toll-like receptor Human genes 0.000 description 7
- 108020000411 Toll-like receptor Proteins 0.000 description 7
- 102100039390 Toll-like receptor 7 Human genes 0.000 description 7
- 230000004913 activation Effects 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 230000004663 cell proliferation Effects 0.000 description 7
- 238000004440 column chromatography Methods 0.000 description 7
- 229940079593 drug Drugs 0.000 description 7
- 239000000706 filtrate Substances 0.000 description 7
- 229910052740 iodine Inorganic materials 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- 238000009097 single-agent therapy Methods 0.000 description 7
- DFVRUHANEXOZGT-UHFFFAOYSA-N tert-butyl n-methyl-n-[2-(methylamino)ethyl]carbamate Chemical compound CNCCN(C)C(=O)OC(C)(C)C DFVRUHANEXOZGT-UHFFFAOYSA-N 0.000 description 7
- 229940044616 toll-like receptor 7 agonist Drugs 0.000 description 7
- XOBKVGSRDJEGRE-UHFFFAOYSA-N 2-(methylamino)ethyl N-butyl-N-methylcarbamate hydrochloride Chemical compound Cl.CCCCN(C)C(=O)OCCNC XOBKVGSRDJEGRE-UHFFFAOYSA-N 0.000 description 6
- XWEFHMKUFKUSBA-UHFFFAOYSA-N 2-(methylamino)ethyl N-methyl-N-propylcarbamate hydrochloride Chemical compound Cl.CCCN(C)C(=O)OCCNC XWEFHMKUFKUSBA-UHFFFAOYSA-N 0.000 description 6
- MCCNKYDFLRPYAH-UHFFFAOYSA-N 2-(methylamino)ethyl pyrrolidine-1-carboxylate hydrochloride Chemical compound Cl.CNCCOC(=O)N1CCCC1 MCCNKYDFLRPYAH-UHFFFAOYSA-N 0.000 description 6
- NHQDETIJWKXCTC-UHFFFAOYSA-N 3-chloroperbenzoic acid Chemical compound OOC(=O)C1=CC=CC(Cl)=C1 NHQDETIJWKXCTC-UHFFFAOYSA-N 0.000 description 6
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 6
- 102000008203 CTLA-4 Antigen Human genes 0.000 description 6
- 229940045513 CTLA4 antagonist Drugs 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 6
- DTLJDNBHCIBSGX-UHFFFAOYSA-N N-methyl-N-[2-(methylamino)ethyl]acetamide hydrochloride Chemical compound Cl.CNCCN(C)C(C)=O DTLJDNBHCIBSGX-UHFFFAOYSA-N 0.000 description 6
- 102100024216 Programmed cell death 1 ligand 1 Human genes 0.000 description 6
- 238000003556 assay Methods 0.000 description 6
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 6
- 210000004443 dendritic cell Anatomy 0.000 description 6
- KHRLPZJTPHCMSQ-YFKPBYRVSA-N ethyl (2s)-2-(methylamino)propanoate Chemical compound CCOC(=O)[C@H](C)NC KHRLPZJTPHCMSQ-YFKPBYRVSA-N 0.000 description 6
- 238000001914 filtration Methods 0.000 description 6
- 238000009472 formulation Methods 0.000 description 6
- 239000008103 glucose Substances 0.000 description 6
- 230000002401 inhibitory effect Effects 0.000 description 6
- 230000003993 interaction Effects 0.000 description 6
- 108010082117 matrigel Proteins 0.000 description 6
- 230000002503 metabolic effect Effects 0.000 description 6
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 6
- 238000003305 oral gavage Methods 0.000 description 6
- 239000000546 pharmaceutical excipient Substances 0.000 description 6
- 239000002244 precipitate Substances 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000004467 single crystal X-ray diffraction Methods 0.000 description 6
- 229940054269 sodium pyruvate Drugs 0.000 description 6
- UZKARLOPBFXQHH-LBPRGKRZSA-N tert-butyl (2s)-2-(methylamino)-3-phenylpropanoate Chemical compound CC(C)(C)OC(=O)[C@@H](NC)CC1=CC=CC=C1 UZKARLOPBFXQHH-LBPRGKRZSA-N 0.000 description 6
- QFMOPRNLDIPINY-UHFFFAOYSA-N tert-butyl 3-(methylamino)propanoate Chemical compound CNCCC(=O)OC(C)(C)C QFMOPRNLDIPINY-UHFFFAOYSA-N 0.000 description 6
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 description 6
- 125000004454 (C1-C6) alkoxycarbonyl group Chemical group 0.000 description 5
- CYNYIHKIEHGYOZ-UHFFFAOYSA-N 1-bromopropane Chemical compound CCCBr CYNYIHKIEHGYOZ-UHFFFAOYSA-N 0.000 description 5
- HFHWUUHRHQXHCU-UHFFFAOYSA-N 4-amino-3-[(4-bromophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile Chemical compound NC=1N(C(NC=1C#N)=O)CC1=CC=C(C=C1)Br HFHWUUHRHQXHCU-UHFFFAOYSA-N 0.000 description 5
- SAZGJPXEDWRWQT-UHFFFAOYSA-N 4-amino-3-[(4-chlorophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile Chemical compound NC=1N(C(NC=1C#N)=O)CC1=CC=C(C=C1)Cl SAZGJPXEDWRWQT-UHFFFAOYSA-N 0.000 description 5
- 229960000549 4-dimethylaminophenol Drugs 0.000 description 5
- WXIMAYUYXNURSS-UHFFFAOYSA-N 6-amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound NC1=C2N(C(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)C(=O)N(CC)CCCC WXIMAYUYXNURSS-UHFFFAOYSA-N 0.000 description 5
- MIOZVXHNRGUNCK-UHFFFAOYSA-N 6-chloro-4-N-[(4-methylphenyl)methyl]-2-propylsulfanylpyrimidine-4,5-diamine Chemical compound ClC1=C(C(=NC(=N1)SCCC)NCC1=CC=C(C=C1)C)N MIOZVXHNRGUNCK-UHFFFAOYSA-N 0.000 description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- OIFBSDVPJOWBCH-UHFFFAOYSA-N Diethyl carbonate Chemical compound CCOC(=O)OCC OIFBSDVPJOWBCH-UHFFFAOYSA-N 0.000 description 5
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 5
- 101100501444 Escherichia phage P1 17 gene Proteins 0.000 description 5
- 101000600434 Homo sapiens Putative uncharacterized protein encoded by MIR7-3HG Proteins 0.000 description 5
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 5
- BZLVMXJERCGZMT-UHFFFAOYSA-N Methyl tert-butyl ether Chemical compound COC(C)(C)C BZLVMXJERCGZMT-UHFFFAOYSA-N 0.000 description 5
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 5
- OPKOKAMJFNKNAS-UHFFFAOYSA-N N-methylethanolamine Chemical compound CNCCO OPKOKAMJFNKNAS-UHFFFAOYSA-N 0.000 description 5
- 102100037401 Putative uncharacterized protein encoded by MIR7-3HG Human genes 0.000 description 5
- AOBORMOPSGHCAX-UHFFFAOYSA-N Tocophersolan Chemical compound OCCOC(=O)CCC(=O)OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C AOBORMOPSGHCAX-UHFFFAOYSA-N 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 5
- 230000016396 cytokine production Effects 0.000 description 5
- BNERQPOWQWDJLR-UHFFFAOYSA-N ethyl n-methyl-n-[2-(methylamino)ethyl]carbamate Chemical compound CCOC(=O)N(C)CCNC BNERQPOWQWDJLR-UHFFFAOYSA-N 0.000 description 5
- 210000003494 hepatocyte Anatomy 0.000 description 5
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 5
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 5
- 238000003384 imaging method Methods 0.000 description 5
- YDNLNVZZTACNJX-UHFFFAOYSA-N isocyanatomethylbenzene Chemical compound O=C=NCC1=CC=CC=C1 YDNLNVZZTACNJX-UHFFFAOYSA-N 0.000 description 5
- 239000003446 ligand Substances 0.000 description 5
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 5
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 5
- 229960002216 methylparaben Drugs 0.000 description 5
- 230000035772 mutation Effects 0.000 description 5
- REBQDPZIZADSPQ-FVGYRXGTSA-N propan-2-yl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride Chemical compound Cl.CN[C@@H](CC(C)C)C(=O)OC(C)C REBQDPZIZADSPQ-FVGYRXGTSA-N 0.000 description 5
- 235000010232 propyl p-hydroxybenzoate Nutrition 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- KDYFGRWQOYBRFD-UHFFFAOYSA-L succinate(2-) Chemical compound [O-]C(=O)CCC([O-])=O KDYFGRWQOYBRFD-UHFFFAOYSA-L 0.000 description 5
- YUFWGGKEIRULTL-VIFPVBQESA-N tert-butyl (2s)-4-methyl-2-(methylamino)pentanoate Chemical compound CC(C)C[C@H](NC)C(=O)OC(C)(C)C YUFWGGKEIRULTL-VIFPVBQESA-N 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 230000003442 weekly effect Effects 0.000 description 5
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 description 5
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 4
- XZORQADPEUSNQJ-UHFFFAOYSA-N 2-(3-piperidin-4-yloxy-1-benzothiophen-2-yl)-5-[(1,3,5-trimethylpyrazol-4-yl)methyl]-1,3,4-oxadiazole Chemical compound CC1=NN(C)C(C)=C1CC1=NN=C(C2=C(C3=CC=CC=C3S2)OC2CCNCC2)O1 XZORQADPEUSNQJ-UHFFFAOYSA-N 0.000 description 4
- UNSHMXUHOHBLIQ-UHFFFAOYSA-N 3-[4-chloro-3-(2-methylphenoxy)naphthalen-1-yl]-6-(trifluoromethyl)-1H-pyrimidine-2,4-dione Chemical compound ClC1=C(C=C(C2=CC=CC=C12)N1C(NC(=CC1=O)C(F)(F)F)=O)OC1=C(C=CC=C1)C UNSHMXUHOHBLIQ-UHFFFAOYSA-N 0.000 description 4
- JHNLZOVBAQWGQU-UHFFFAOYSA-N 380814_sial Chemical compound CS(O)(=O)=O.O=P(=O)OP(=O)=O JHNLZOVBAQWGQU-UHFFFAOYSA-N 0.000 description 4
- 125000004172 4-methoxyphenyl group Chemical group [H]C1=C([H])C(OC([H])([H])[H])=C([H])C([H])=C1* 0.000 description 4
- 238000011814 C57BL/6N mouse Methods 0.000 description 4
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 4
- 108090000695 Cytokines Proteins 0.000 description 4
- VQTUBCCKSQIDNK-UHFFFAOYSA-N Isobutene Chemical compound CC(C)=C VQTUBCCKSQIDNK-UHFFFAOYSA-N 0.000 description 4
- AFVFQIVMOAPDHO-UHFFFAOYSA-N Methanesulfonic acid Chemical compound CS(O)(=O)=O AFVFQIVMOAPDHO-UHFFFAOYSA-N 0.000 description 4
- 102000010168 Myeloid Differentiation Factor 88 Human genes 0.000 description 4
- 108010077432 Myeloid Differentiation Factor 88 Proteins 0.000 description 4
- KISZAGQTIXIVAR-UHFFFAOYSA-N OC(=O)c1ccc2c(CCCC(c3ccc(Cl)cc3Cl)=C2c2ccc(OC3CCN(CCCF)C3)cc2)c1 Chemical compound OC(=O)c1ccc2c(CCCC(c3ccc(Cl)cc3Cl)=C2c2ccc(OC3CCN(CCCF)C3)cc2)c1 KISZAGQTIXIVAR-UHFFFAOYSA-N 0.000 description 4
- 239000004793 Polystyrene Substances 0.000 description 4
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 4
- 239000011324 bead Substances 0.000 description 4
- 210000001715 carotid artery Anatomy 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 229940126209 compound 43b Drugs 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- UNHAQFDONQNXAQ-FJXQXJEOSA-N ethyl (2s)-3-methyl-2-(methylamino)butanoate;hydrochloride Chemical compound Cl.CCOC(=O)[C@@H](NC)C(C)C UNHAQFDONQNXAQ-FJXQXJEOSA-N 0.000 description 4
- KFZPMSMKKYNZTC-UHFFFAOYSA-N ethyl 2-(methylamino)ethyl carbonate;hydrochloride Chemical compound Cl.CCOC(=O)OCCNC KFZPMSMKKYNZTC-UHFFFAOYSA-N 0.000 description 4
- 238000007866 imination reaction Methods 0.000 description 4
- 230000036039 immunity Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000007912 intraperitoneal administration Methods 0.000 description 4
- 238000001990 intravenous administration Methods 0.000 description 4
- HVTICUPFWKNHNG-UHFFFAOYSA-N iodoethane Chemical compound CCI HVTICUPFWKNHNG-UHFFFAOYSA-N 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- JFOZKMSJYSPYLN-QHCPKHFHSA-N lifitegrast Chemical compound CS(=O)(=O)C1=CC=CC(C[C@H](NC(=O)C=2C(=C3CCN(CC3=CC=2Cl)C(=O)C=2C=C3OC=CC3=CC=2)Cl)C(O)=O)=C1 JFOZKMSJYSPYLN-QHCPKHFHSA-N 0.000 description 4
- 210000002540 macrophage Anatomy 0.000 description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 4
- CXHHBNMLPJOKQD-UHFFFAOYSA-M methyl carbonate Chemical compound COC([O-])=O CXHHBNMLPJOKQD-UHFFFAOYSA-M 0.000 description 4
- 125000000250 methylamino group Chemical group [H]N(*)C([H])([H])[H] 0.000 description 4
- UQRZZODUMPZZEB-UHFFFAOYSA-N n-(2-methoxyethyl)-n-methylcarbamoyl chloride Chemical compound COCCN(C)C(Cl)=O UQRZZODUMPZZEB-UHFFFAOYSA-N 0.000 description 4
- SKEAGJWUKCNICJ-LSDHHAIUSA-N n-[(4-fluoro-3-methoxyphenyl)methyl]-6-[2-[[(2s,5r)-5-(hydroxymethyl)-1,4-dioxan-2-yl]methyl]tetrazol-5-yl]-2-methylpyrimidine-4-carboxamide Chemical compound C1=C(F)C(OC)=CC(CNC(=O)C=2N=C(C)N=C(C=2)C2=NN(C[C@@H]3OC[C@@H](CO)OC3)N=N2)=C1 SKEAGJWUKCNICJ-LSDHHAIUSA-N 0.000 description 4
- JYRMPDXSENSKDY-UHFFFAOYSA-N n-ethyl-n-(2-methoxyethyl)carbamoyl chloride Chemical compound CCN(C(Cl)=O)CCOC JYRMPDXSENSKDY-UHFFFAOYSA-N 0.000 description 4
- 210000005259 peripheral blood Anatomy 0.000 description 4
- 239000011886 peripheral blood Substances 0.000 description 4
- 229940113116 polyethylene glycol 1000 Drugs 0.000 description 4
- 229920002223 polystyrene Polymers 0.000 description 4
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 description 4
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 4
- 125000006413 ring segment Chemical group 0.000 description 4
- 239000000741 silica gel Substances 0.000 description 4
- 229910002027 silica gel Inorganic materials 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 238000007920 subcutaneous administration Methods 0.000 description 4
- 238000001356 surgical procedure Methods 0.000 description 4
- 239000000725 suspension Substances 0.000 description 4
- LMBFAGIMSUYTBN-MPZNNTNKSA-N teixobactin Chemical compound C([C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](CCC(N)=O)C(=O)N[C@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H]1C(N[C@@H](C)C(=O)N[C@@H](C[C@@H]2NC(=N)NC2)C(=O)N[C@H](C(=O)O[C@H]1C)[C@@H](C)CC)=O)NC)C1=CC=CC=C1 LMBFAGIMSUYTBN-MPZNNTNKSA-N 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 3
- PPAULTVPKLVLII-UHFFFAOYSA-N 4,5-diaminopyrimidine Chemical compound NC1=CN=CN=C1N PPAULTVPKLVLII-UHFFFAOYSA-N 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 3
- VFRQJCXMZAJFNM-UHFFFAOYSA-N 4-amino-3-[(4-methylphenyl)methyl]-2-oxo-1h-imidazole-5-carbonitrile Chemical compound C1=CC(C)=CC=C1CN1C(=O)NC(C#N)=C1N VFRQJCXMZAJFNM-UHFFFAOYSA-N 0.000 description 3
- QYTROLFWFOAQLB-YTTGMZPUSA-N 6-amino-9-[(4-methylphenyl)methyl]-2-(propylsulfonimidoyl)-7-(pyrrolidine-1-carbonyl)purin-8-one Chemical compound CCC[S@](=N)(=O)C1=NC(N)=C2N(C(=O)N3CCCC3)C(=O)N(CC3=CC=C(C)C=C3)C2=N1 QYTROLFWFOAQLB-YTTGMZPUSA-N 0.000 description 3
- FNUMFDZCSMALPX-XIFFEERXSA-N 6-amino-N-butyl-9-[(4-chlorophenyl)methyl]-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound CCCCN(C)C(=O)N1C(=O)N(CC2=CC=C(Cl)C=C2)C2=NC(=NC(N)=C12)[S@@](=N)(=O)CCC FNUMFDZCSMALPX-XIFFEERXSA-N 0.000 description 3
- NCHPONKCXVTORK-UHFFFAOYSA-N 6-chloro-N-[(4-methylphenyl)methyl]-5-nitro-2-propylsulfanylpyrimidin-4-amine Chemical compound CCCSC1=NC(NCC2=CC=C(C)C=C2)=C(C(Cl)=N1)[N+]([O-])=O NCHPONKCXVTORK-UHFFFAOYSA-N 0.000 description 3
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 3
- 101150041968 CDC13 gene Proteins 0.000 description 3
- CKDWPUIZGOQOOM-UHFFFAOYSA-N Carbamyl chloride Chemical compound NC(Cl)=O CKDWPUIZGOQOOM-UHFFFAOYSA-N 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 102100037850 Interferon gamma Human genes 0.000 description 3
- 108010074328 Interferon-gamma Proteins 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- KDYFGRWQOYBRFD-UHFFFAOYSA-N Succinic acid Natural products OC(=O)CCC(O)=O KDYFGRWQOYBRFD-UHFFFAOYSA-N 0.000 description 3
- 230000006052 T cell proliferation Effects 0.000 description 3
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 3
- 239000000556 agonist Substances 0.000 description 3
- 230000001270 agonistic effect Effects 0.000 description 3
- LJPIQCGBOGKQOK-UHFFFAOYSA-N azetidine-1-carbonyl chloride Chemical compound ClC(=O)N1CCC1 LJPIQCGBOGKQOK-UHFFFAOYSA-N 0.000 description 3
- 210000002469 basement membrane Anatomy 0.000 description 3
- 210000001185 bone marrow Anatomy 0.000 description 3
- PFKFTWBEEFSNDU-UHFFFAOYSA-N carbonyldiimidazole Chemical compound C1=CN=CN1C(=O)N1C=CN=C1 PFKFTWBEEFSNDU-UHFFFAOYSA-N 0.000 description 3
- 231100000504 carcinogenesis Toxicity 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 230000001086 cytosolic effect Effects 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- ONRCQWWOUUVQMT-JEDNCBNOSA-N ethyl (2s)-2-(methylamino)propanoate;hydrochloride Chemical compound Cl.CCOC(=O)[C@H](C)NC ONRCQWWOUUVQMT-JEDNCBNOSA-N 0.000 description 3
- OKZXFBDHFSQMMW-QRPNPIFTSA-N ethyl (2s)-4-methyl-2-(methylamino)pentanoate;hydrochloride Chemical compound Cl.CCOC(=O)[C@@H](NC)CC(C)C OKZXFBDHFSQMMW-QRPNPIFTSA-N 0.000 description 3
- 238000003818 flash chromatography Methods 0.000 description 3
- 235000019253 formic acid Nutrition 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 210000002865 immune cell Anatomy 0.000 description 3
- 238000001802 infusion Methods 0.000 description 3
- 150000007529 inorganic bases Chemical class 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 210000000265 leukocyte Anatomy 0.000 description 3
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 3
- 239000011159 matrix material Substances 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- GZNVFRIHZRYMFK-UHFFFAOYSA-N n-butyl-n-ethylcarbamoyl chloride Chemical compound CCCCN(CC)C(Cl)=O GZNVFRIHZRYMFK-UHFFFAOYSA-N 0.000 description 3
- CHGVIQWWXDOWRN-UHFFFAOYSA-N n-butyl-n-methylcarbamoyl chloride Chemical compound CCCCN(C)C(Cl)=O CHGVIQWWXDOWRN-UHFFFAOYSA-N 0.000 description 3
- ZTBGJRUHDDPDEK-UHFFFAOYSA-N n-ethyl-n-propylcarbamoyl chloride Chemical compound CCCN(CC)C(Cl)=O ZTBGJRUHDDPDEK-UHFFFAOYSA-N 0.000 description 3
- BWOVQCBKUXJWNF-UHFFFAOYSA-N n-methyl-n-(2-methylpropyl)carbamoyl chloride Chemical compound CC(C)CN(C)C(Cl)=O BWOVQCBKUXJWNF-UHFFFAOYSA-N 0.000 description 3
- VMLZTGZTAWBKSP-UHFFFAOYSA-N n-methyl-n-propan-2-ylcarbamoyl chloride Chemical compound CC(C)N(C)C(Cl)=O VMLZTGZTAWBKSP-UHFFFAOYSA-N 0.000 description 3
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 3
- 230000036470 plasma concentration Effects 0.000 description 3
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 3
- 238000002953 preparative HPLC Methods 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000002062 proliferating effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 238000007363 ring formation reaction Methods 0.000 description 3
- WBHQBSYUUJJSRZ-UHFFFAOYSA-M sodium bisulfate Chemical compound [Na+].OS([O-])(=O)=O WBHQBSYUUJJSRZ-UHFFFAOYSA-M 0.000 description 3
- 229910000342 sodium bisulfate Inorganic materials 0.000 description 3
- 230000000638 stimulation Effects 0.000 description 3
- 239000000758 substrate Substances 0.000 description 3
- CXRZBVYXFWLYLK-LBPRGKRZSA-N tert-butyl (2S)-2-[carbonochloridoyl(methyl)amino]-3-phenylpropanoate Chemical compound ClC(=O)N([C@H](C(=O)OC(C)(C)C)CC1=CC=CC=C1)C CXRZBVYXFWLYLK-LBPRGKRZSA-N 0.000 description 3
- UCPYLLCMEDAXFR-UHFFFAOYSA-N triphosgene Chemical compound ClC(Cl)(Cl)OC(=O)OC(Cl)(Cl)Cl UCPYLLCMEDAXFR-UHFFFAOYSA-N 0.000 description 3
- 241000701161 unidentified adenovirus Species 0.000 description 3
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 3
- 239000012224 working solution Substances 0.000 description 3
- 239000011701 zinc Substances 0.000 description 3
- QBYIENPQHBMVBV-HFEGYEGKSA-N (2R)-2-hydroxy-2-phenylacetic acid Chemical compound O[C@@H](C(O)=O)c1ccccc1.O[C@@H](C(O)=O)c1ccccc1 QBYIENPQHBMVBV-HFEGYEGKSA-N 0.000 description 2
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 2
- 102000040650 (ribonucleotides)n+m Human genes 0.000 description 2
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 2
- IDPURXSQCKYKIJ-UHFFFAOYSA-N 1-(4-methoxyphenyl)methanamine Chemical compound COC1=CC=C(CN)C=C1 IDPURXSQCKYKIJ-UHFFFAOYSA-N 0.000 description 2
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- UPMKNNFFRWACOU-UHFFFAOYSA-N 2-[carbonochloridoyl(methyl)amino]ethyl N-butyl-N-methylcarbamate Chemical compound C(CCC)N(C(OCCN(C)C(=O)Cl)=O)C UPMKNNFFRWACOU-UHFFFAOYSA-N 0.000 description 2
- NOHWPMMRYDCMRU-UHFFFAOYSA-N 2-[carbonochloridoyl(methyl)amino]ethyl N-methyl-N-propylcarbamate Chemical compound CN(C(OCCN(C)C(=O)Cl)=O)CCC NOHWPMMRYDCMRU-UHFFFAOYSA-N 0.000 description 2
- YPZUOQOTBSMJNT-UHFFFAOYSA-N 2-[carbonochloridoyl(methyl)amino]ethyl ethyl carbonate Chemical compound CCOC(=O)OCCN(C)C(Cl)=O YPZUOQOTBSMJNT-UHFFFAOYSA-N 0.000 description 2
- JBUSJTBITAJTQT-UHFFFAOYSA-N 2-[carbonochloridoyl(methyl)amino]ethyl pyrrolidine-1-carboxylate Chemical compound N1(CCCC1)C(=O)OCCN(C)C(=O)Cl JBUSJTBITAJTQT-UHFFFAOYSA-N 0.000 description 2
- OZYZGGNBOBFYNI-UHFFFAOYSA-N 2-[methyl-[(2-methylpropan-2-yl)oxycarbonyl]amino]ethyl pyrrolidine-1-carboxylate Chemical compound N1(CCCC1)C(=O)OCCN(C)C(=O)OC(C)(C)C OZYZGGNBOBFYNI-UHFFFAOYSA-N 0.000 description 2
- MEUWQVWJLLBVQI-UHFFFAOYSA-N 2-aminopropanedinitrile;4-methylbenzenesulfonic acid Chemical compound N#CC([NH3+])C#N.CC1=CC=C(S([O-])(=O)=O)C=C1 MEUWQVWJLLBVQI-UHFFFAOYSA-N 0.000 description 2
- VFOKSTCIRGDTBR-UHFFFAOYSA-N 4-amino-2-butoxy-8-[[3-(pyrrolidin-1-ylmethyl)phenyl]methyl]-5,7-dihydropteridin-6-one Chemical compound C12=NC(OCCCC)=NC(N)=C2NC(=O)CN1CC(C=1)=CC=CC=1CN1CCCC1 VFOKSTCIRGDTBR-UHFFFAOYSA-N 0.000 description 2
- QYTROLFWFOAQLB-UHFFFAOYSA-N 6-amino-9-[(4-methylphenyl)methyl]-2-(propylsulfonimidoyl)-7-(pyrrolidine-1-carbonyl)purin-8-one Chemical compound CCCS(=N)(=O)c1nc(N)c2n(C(=O)N3CCCC3)c(=O)n(Cc3ccc(C)cc3)c2n1 QYTROLFWFOAQLB-UHFFFAOYSA-N 0.000 description 2
- PNGYMTFMJSIZHE-UHFFFAOYSA-N 6-amino-9-benzyl-N-methyl-N-(2-methylpropyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound NC1=C2N(C(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)C(=O)N(C)CC(C)C PNGYMTFMJSIZHE-UHFFFAOYSA-N 0.000 description 2
- HQXUZKDMNVTFOP-XIFFEERXSA-N 6-amino-N-(2-methoxyethyl)-N-methyl-9-[(4-methylphenyl)methyl]-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound CCC[S@](=N)(=O)C1=NC(N)=C2N(C(=O)N(C)CCOC)C(=O)N(CC3=CC=C(C)C=C3)C2=N1 HQXUZKDMNVTFOP-XIFFEERXSA-N 0.000 description 2
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- 208000005623 Carcinogenesis Diseases 0.000 description 2
- 108010051219 Cre recombinase Proteins 0.000 description 2
- RGHNJXZEOKUKBD-SQOUGZDYSA-N D-gluconic acid Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C(O)=O RGHNJXZEOKUKBD-SQOUGZDYSA-N 0.000 description 2
- ODBLHEXUDAPZAU-ZAFYKAAXSA-N D-threo-isocitric acid Chemical compound OC(=O)[C@H](O)[C@@H](C(O)=O)CC(O)=O ODBLHEXUDAPZAU-ZAFYKAAXSA-N 0.000 description 2
- FEWJPZIEWOKRBE-JCYAYHJZSA-N Dextrotartaric acid Chemical compound OC(=O)[C@H](O)[C@@H](O)C(O)=O FEWJPZIEWOKRBE-JCYAYHJZSA-N 0.000 description 2
- 108700024394 Exon Proteins 0.000 description 2
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 description 2
- 229940126656 GS-4224 Drugs 0.000 description 2
- 239000001828 Gelatine Substances 0.000 description 2
- AEMRFAOFKBGASW-UHFFFAOYSA-N Glycolic acid Chemical compound OCC(O)=O AEMRFAOFKBGASW-UHFFFAOYSA-N 0.000 description 2
- 239000007821 HATU Substances 0.000 description 2
- 208000005176 Hepatitis C Diseases 0.000 description 2
- 102000007576 Interferon Regulatory Factor-7 Human genes 0.000 description 2
- 108010032036 Interferon Regulatory Factor-7 Proteins 0.000 description 2
- 108010063738 Interleukins Proteins 0.000 description 2
- 102000015696 Interleukins Human genes 0.000 description 2
- 101710088105 Isocitrate dehydrogenase [NAD] subunit 1, mitochondrial Proteins 0.000 description 2
- 101710086399 Isocitrate dehydrogenase [NAD] subunit 2, mitochondrial Proteins 0.000 description 2
- 102100021332 Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial Human genes 0.000 description 2
- ODBLHEXUDAPZAU-FONMRSAGSA-N Isocitric acid Natural products OC(=O)[C@@H](O)[C@H](C(O)=O)CC(O)=O ODBLHEXUDAPZAU-FONMRSAGSA-N 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 108091007960 PI3Ks Proteins 0.000 description 2
- 102100032543 Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Human genes 0.000 description 2
- 101710132081 Phosphatidylinositol 3,4,5-trisphosphate 3-phosphatase and dual-specificity protein phosphatase PTEN Proteins 0.000 description 2
- 102000003993 Phosphatidylinositol 3-kinases Human genes 0.000 description 2
- 108090000430 Phosphatidylinositol 3-kinases Proteins 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 239000004698 Polyethylene Substances 0.000 description 2
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 description 2
- IWYDHOAUDWTVEP-UHFFFAOYSA-N R-2-phenyl-2-hydroxyacetic acid Natural products OC(=O)C(O)C1=CC=CC=C1 IWYDHOAUDWTVEP-UHFFFAOYSA-N 0.000 description 2
- 108010060825 Toll-Like Receptor 7 Proteins 0.000 description 2
- 102000008236 Toll-Like Receptor 7 Human genes 0.000 description 2
- 108010018242 Transcription Factor AP-1 Proteins 0.000 description 2
- 102000040945 Transcription factor Human genes 0.000 description 2
- 108091023040 Transcription factor Proteins 0.000 description 2
- 102100023132 Transcription factor Jun Human genes 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102100040247 Tumor necrosis factor Human genes 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 210000000683 abdominal cavity Anatomy 0.000 description 2
- 230000003187 abdominal effect Effects 0.000 description 2
- 210000003815 abdominal wall Anatomy 0.000 description 2
- 230000003213 activating effect Effects 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 230000008484 agonism Effects 0.000 description 2
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 239000005557 antagonist Substances 0.000 description 2
- RWZYAGGXGHYGMB-UHFFFAOYSA-N anthranilic acid Chemical compound NC1=CC=CC=C1C(O)=O RWZYAGGXGHYGMB-UHFFFAOYSA-N 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 229940127219 anticoagulant drug Drugs 0.000 description 2
- 230000005784 autoimmunity Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- CPEKAXYCDKETEN-UHFFFAOYSA-N benzoyl isothiocyanate Chemical compound S=C=NC(=O)C1=CC=CC=C1 CPEKAXYCDKETEN-UHFFFAOYSA-N 0.000 description 2
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 description 2
- 238000011953 bioanalysis Methods 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000001772 blood platelet Anatomy 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- FJDQFPXHSGXQBY-UHFFFAOYSA-L caesium carbonate Chemical compound [Cs+].[Cs+].[O-]C([O-])=O FJDQFPXHSGXQBY-UHFFFAOYSA-L 0.000 description 2
- RYYVLZVUVIJVGH-UHFFFAOYSA-N caffeine Chemical compound CN1C(=O)N(C)C(=O)C2=C1N=CN2C RYYVLZVUVIJVGH-UHFFFAOYSA-N 0.000 description 2
- 238000011088 calibration curve Methods 0.000 description 2
- 230000036952 cancer formation Effects 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 210000004534 cecum Anatomy 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 235000015165 citric acid Nutrition 0.000 description 2
- 229940125904 compound 1 Drugs 0.000 description 2
- 238000012937 correction Methods 0.000 description 2
- 230000004940 costimulation Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 238000012377 drug delivery Methods 0.000 description 2
- 210000001842 enterocyte Anatomy 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- OHGJZKDJUOSKEX-UHFFFAOYSA-N ethyl 2-[carbonochloridoyl(methyl)amino]acetate Chemical compound C(=O)(OCC)CN(C(=O)Cl)C OHGJZKDJUOSKEX-UHFFFAOYSA-N 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 230000008020 evaporation Effects 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 235000003599 food sweetener Nutrition 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- KWIUHFFTVRNATP-UHFFFAOYSA-N glycine betaine Chemical compound C[N+](C)(C)CC([O-])=O KWIUHFFTVRNATP-UHFFFAOYSA-N 0.000 description 2
- 229940093915 gynecological organic acid Drugs 0.000 description 2
- 208000002672 hepatitis B Diseases 0.000 description 2
- 125000005842 heteroatom Chemical group 0.000 description 2
- 210000003630 histaminocyte Anatomy 0.000 description 2
- 102000045715 human TLR7 Human genes 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 230000001900 immune effect Effects 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 230000000977 initiatory effect Effects 0.000 description 2
- 230000015788 innate immune response Effects 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000001325 log-rank test Methods 0.000 description 2
- 235000019359 magnesium stearate Nutrition 0.000 description 2
- 239000001630 malic acid Substances 0.000 description 2
- 235000011090 malic acid Nutrition 0.000 description 2
- 229960002510 mandelic acid Drugs 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- NHZHZPNZWCILQK-UHFFFAOYSA-N methyl 2-[methyl-[(2-methylpropan-2-yl)oxycarbonyl]amino]ethyl carbonate Chemical compound C(OCCN(C)C(=O)OC(C)(C)C)(OC)=O NHZHZPNZWCILQK-UHFFFAOYSA-N 0.000 description 2
- XMJHPCRAQCTCFT-UHFFFAOYSA-N methyl chloroformate Chemical compound COC(Cl)=O XMJHPCRAQCTCFT-UHFFFAOYSA-N 0.000 description 2
- CIHAWTDTDVMLKM-UHFFFAOYSA-N methyl n-methyl-n-[2-(methylamino)ethyl]carbamate Chemical compound CNCCN(C)C(=O)OC CIHAWTDTDVMLKM-UHFFFAOYSA-N 0.000 description 2
- BSCFAUJKRHSBGE-UHFFFAOYSA-N methyl(propyl)carbamic acid Chemical compound CCCN(C)C(O)=O BSCFAUJKRHSBGE-UHFFFAOYSA-N 0.000 description 2
- 210000001589 microsome Anatomy 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 210000001616 monocyte Anatomy 0.000 description 2
- 210000000066 myeloid cell Anatomy 0.000 description 2
- ZHNYOUMPJQCUET-UHFFFAOYSA-N n,n-bis(2-methoxyethyl)carbamoyl chloride Chemical compound COCCN(C(Cl)=O)CCOC ZHNYOUMPJQCUET-UHFFFAOYSA-N 0.000 description 2
- CYSWHFKPBBXLKH-UHFFFAOYSA-N n-(2-methoxyethyl)-n-propylcarbamoyl chloride Chemical compound CCCN(C(Cl)=O)CCOC CYSWHFKPBBXLKH-UHFFFAOYSA-N 0.000 description 2
- 125000004123 n-propyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])* 0.000 description 2
- 229940080607 nexavar Drugs 0.000 description 2
- 210000004967 non-hematopoietic stem cell Anatomy 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 125000001400 nonyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 2
- 238000001543 one-way ANOVA Methods 0.000 description 2
- 150000007524 organic acids Chemical class 0.000 description 2
- 235000005985 organic acids Nutrition 0.000 description 2
- 239000012074 organic phase Substances 0.000 description 2
- 239000007800 oxidant agent Substances 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000037361 pathway Effects 0.000 description 2
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 2
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 2
- 239000000825 pharmaceutical preparation Substances 0.000 description 2
- 239000012071 phase Substances 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 230000000704 physical effect Effects 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 229910000027 potassium carbonate Inorganic materials 0.000 description 2
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 2
- 239000004405 propyl p-hydroxybenzoate Substances 0.000 description 2
- 229960003415 propylparaben Drugs 0.000 description 2
- 238000000159 protein binding assay Methods 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- XACWJIQLDLUFSR-UHFFFAOYSA-N pyrrolidine-1-carbonyl chloride Chemical compound ClC(=O)N1CCCC1 XACWJIQLDLUFSR-UHFFFAOYSA-N 0.000 description 2
- 238000007674 radiofrequency ablation Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 230000000284 resting effect Effects 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 230000019491 signal transduction Effects 0.000 description 2
- 150000003384 small molecules Chemical class 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 2
- 239000003765 sweetening agent Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- FJVVQPOMFUHNBK-UHFFFAOYSA-N tert-butyl 3-[carbonochloridoyl(methyl)amino]propanoate Chemical compound ClC(=O)N(CCC(=O)OC(C)(C)C)C FJVVQPOMFUHNBK-UHFFFAOYSA-N 0.000 description 2
- HNEUIHISGSLDKO-UHFFFAOYSA-N tert-butyl N-methyl-N-[2-[methyl(propyl)carbamoyl]oxyethyl]carbamate Chemical compound CCCN(C)C(=O)OCCN(C)C(=O)OC(C)(C)C HNEUIHISGSLDKO-UHFFFAOYSA-N 0.000 description 2
- YAPQBXQYLJRXSA-UHFFFAOYSA-N theobromine Chemical compound CN1C(=O)NC(=O)C2=C1N=CN2C YAPQBXQYLJRXSA-UHFFFAOYSA-N 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- ODBLHEXUDAPZAU-UHFFFAOYSA-N threo-D-isocitric acid Natural products OC(=O)C(O)C(C(O)=O)CC(O)=O ODBLHEXUDAPZAU-UHFFFAOYSA-N 0.000 description 2
- JOXIMZWYDAKGHI-UHFFFAOYSA-N toluene-4-sulfonic acid Chemical compound CC1=CC=C(S(O)(=O)=O)C=C1 JOXIMZWYDAKGHI-UHFFFAOYSA-N 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 description 2
- 238000011830 transgenic mouse model Methods 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 229940086542 triethylamine Drugs 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 description 2
- 239000005483 tyrosine kinase inhibitor Substances 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 229950003036 vesatolimod Drugs 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- XJODGRWDFZVTKW-LURJTMIESA-N (2s)-4-methyl-2-(methylamino)pentanoic acid Chemical compound CN[C@H](C(O)=O)CC(C)C XJODGRWDFZVTKW-LURJTMIESA-N 0.000 description 1
- QFLWZFQWSBQYPS-AWRAUJHKSA-N (3S)-3-[[(2S)-2-[[(2S)-2-[5-[(3aS,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoylamino]-3-methylbutanoyl]amino]-3-(4-hydroxyphenyl)propanoyl]amino]-4-[1-bis(4-chlorophenoxy)phosphorylbutylamino]-4-oxobutanoic acid Chemical compound CCCC(NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](Cc1ccc(O)cc1)NC(=O)[C@@H](NC(=O)CCCCC1SC[C@@H]2NC(=O)N[C@H]12)C(C)C)P(=O)(Oc1ccc(Cl)cc1)Oc1ccc(Cl)cc1 QFLWZFQWSBQYPS-AWRAUJHKSA-N 0.000 description 1
- HMTSWYPNXFHGEP-UHFFFAOYSA-N (4-methylphenyl)methanamine Chemical compound CC1=CC=C(CN)C=C1 HMTSWYPNXFHGEP-UHFFFAOYSA-N 0.000 description 1
- MIOPJNTWMNEORI-GMSGAONNSA-N (S)-camphorsulfonic acid Chemical compound C1C[C@@]2(CS(O)(=O)=O)C(=O)C[C@@H]1C2(C)C MIOPJNTWMNEORI-GMSGAONNSA-N 0.000 description 1
- JNPGUXGVLNJQSQ-BGGMYYEUSA-M (e,3r,5s)-7-[4-(4-fluorophenyl)-1,2-di(propan-2-yl)pyrrol-3-yl]-3,5-dihydroxyhept-6-enoate Chemical compound CC(C)N1C(C(C)C)=C(\C=C\[C@@H](O)C[C@@H](O)CC([O-])=O)C(C=2C=CC(F)=CC=2)=C1 JNPGUXGVLNJQSQ-BGGMYYEUSA-M 0.000 description 1
- WBYWAXJHAXSJNI-VOTSOKGWSA-M .beta-Phenylacrylic acid Natural products [O-]C(=O)\C=C\C1=CC=CC=C1 WBYWAXJHAXSJNI-VOTSOKGWSA-M 0.000 description 1
- KNYDWDHLGFMGCO-UHFFFAOYSA-N 1-(isocyanatomethyl)-4-methylbenzene Chemical compound CC1=CC=C(CN=C=O)C=C1 KNYDWDHLGFMGCO-UHFFFAOYSA-N 0.000 description 1
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 1
- IXWMDGLNJQNMIO-UHFFFAOYSA-N 1-bromo-4-(isocyanatomethyl)benzene Chemical compound BrC1=CC=C(CN=C=O)C=C1 IXWMDGLNJQNMIO-UHFFFAOYSA-N 0.000 description 1
- OAXSVEFCBLZGCA-UHFFFAOYSA-N 1-chloro-4-(isocyanatomethyl)benzene Chemical compound ClC1=CC=C(CN=C=O)C=C1 OAXSVEFCBLZGCA-UHFFFAOYSA-N 0.000 description 1
- VSTXCZGEEVFJES-UHFFFAOYSA-N 1-cycloundecyl-1,5-diazacycloundec-5-ene Chemical compound C1CCCCCC(CCCC1)N1CCCCCC=NCCC1 VSTXCZGEEVFJES-UHFFFAOYSA-N 0.000 description 1
- CPKVUHPKYQGHMW-UHFFFAOYSA-N 1-ethenylpyrrolidin-2-one;molecular iodine Chemical compound II.C=CN1CCCC1=O CPKVUHPKYQGHMW-UHFFFAOYSA-N 0.000 description 1
- HHSIWJYERNCLKQ-UHFFFAOYSA-N 1-fluoro-4-(isocyanatomethyl)benzene Chemical compound FC1=CC=C(CN=C=O)C=C1 HHSIWJYERNCLKQ-UHFFFAOYSA-N 0.000 description 1
- PNGUXBKJTJSJHK-UHFFFAOYSA-N 2-[methyl-[(2-methylpropan-2-yl)oxycarbonyl]amino]ethyl N,N-diethylcarbamate Chemical compound C(C)(C)(C)OC(=O)N(CCOC(N(CC)CC)=O)C PNGUXBKJTJSJHK-UHFFFAOYSA-N 0.000 description 1
- DSMWZBNKNKCJFE-UHFFFAOYSA-N 2-[methyl-[(2-methylpropan-2-yl)oxycarbonyl]amino]ethyl N-butyl-N-methylcarbamate Chemical compound C(CCC)N(C(OCCN(C)C(=O)OC(C)(C)C)=O)C DSMWZBNKNKCJFE-UHFFFAOYSA-N 0.000 description 1
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- 229940013085 2-diethylaminoethanol Drugs 0.000 description 1
- SGUAFYQXFOLMHL-UHFFFAOYSA-N 2-hydroxy-5-{1-hydroxy-2-[(4-phenylbutan-2-yl)amino]ethyl}benzamide Chemical compound C=1C=C(O)C(C(N)=O)=CC=1C(O)CNC(C)CCC1=CC=CC=C1 SGUAFYQXFOLMHL-UHFFFAOYSA-N 0.000 description 1
- IBZKBSXREAQDTO-UHFFFAOYSA-N 2-methoxy-n-(2-methoxyethyl)ethanamine Chemical compound COCCNCCOC IBZKBSXREAQDTO-UHFFFAOYSA-N 0.000 description 1
- KOHBEDRJXKOYHL-UHFFFAOYSA-N 2-methoxy-n-methylethanamine Chemical compound CNCCOC KOHBEDRJXKOYHL-UHFFFAOYSA-N 0.000 description 1
- WLJVXDMOQOGPHL-PPJXEINESA-N 2-phenylacetic acid Chemical compound O[14C](=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-PPJXEINESA-N 0.000 description 1
- GRUPMMBRLDBTDD-UHFFFAOYSA-N 3-[2-(2-methyl-1,3-thiazol-4-yl)ethynyl]benzonitrile Chemical compound S1C(C)=NC(C#CC=2C=C(C=CC=2)C#N)=C1 GRUPMMBRLDBTDD-UHFFFAOYSA-N 0.000 description 1
- BMYNFMYTOJXKLE-UHFFFAOYSA-N 3-azaniumyl-2-hydroxypropanoate Chemical compound NCC(O)C(O)=O BMYNFMYTOJXKLE-UHFFFAOYSA-N 0.000 description 1
- DDEDQHVHVPJFAC-UHFFFAOYSA-N 4,6-dichloro-5-nitro-2-propylsulfanylpyrimidine Chemical compound CCCSC1=NC(Cl)=C([N+]([O-])=O)C(Cl)=N1 DDEDQHVHVPJFAC-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- FBAIGEMWTOSCRU-UHFFFAOYSA-N 4-methylpiperazine-1-carbonyl chloride Chemical compound CN1CCN(C(Cl)=O)CC1 FBAIGEMWTOSCRU-UHFFFAOYSA-N 0.000 description 1
- YDNSNQRKIINKPV-UHFFFAOYSA-N 4-piperidin-1-ylpiperidine-1-carbonyl chloride Chemical compound C1CN(C(=O)Cl)CCC1N1CCCCC1 YDNSNQRKIINKPV-UHFFFAOYSA-N 0.000 description 1
- YXFJJQUVGAPZKB-UHFFFAOYSA-N 6-amino-9-[(4-methylphenyl)methyl]-2-(propylsulfonimidoyl)-7H-purin-8-one Chemical compound NC1=C2NC(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=C(C=C1)C)=O YXFJJQUVGAPZKB-UHFFFAOYSA-N 0.000 description 1
- LSBVVFLIWUXURT-UHFFFAOYSA-N 6-amino-9-benzyl-N-(2-methoxyethyl)-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound NC1=C2N(C(N(C2=NC(=N1)S(=O)(=N)CCC)CC1=CC=CC=C1)=O)C(=O)N(C)CCOC LSBVVFLIWUXURT-UHFFFAOYSA-N 0.000 description 1
- HQXUZKDMNVTFOP-UHFFFAOYSA-N 6-amino-N-(2-methoxyethyl)-N-methyl-9-[(4-methylphenyl)methyl]-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide Chemical compound CCCS(=N)(=O)c1nc(N)c2n(C(=O)N(C)CCOC)c(=O)n(Cc3ccc(C)cc3)c2n1 HQXUZKDMNVTFOP-UHFFFAOYSA-N 0.000 description 1
- 241000321096 Adenoides Species 0.000 description 1
- 230000007730 Akt signaling Effects 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 108010045634 B7 Antigens Proteins 0.000 description 1
- 102000005738 B7 Antigens Human genes 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 108700031361 Brachyury Proteins 0.000 description 1
- 102100026008 Breakpoint cluster region protein Human genes 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 210000001266 CD8-positive T-lymphocyte Anatomy 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- WBYWAXJHAXSJNI-SREVYHEPSA-N Cinnamic acid Chemical compound OC(=O)\C=C/C1=CC=CC=C1 WBYWAXJHAXSJNI-SREVYHEPSA-N 0.000 description 1
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 108010026925 Cytochrome P-450 CYP2C19 Proteins 0.000 description 1
- 108010000543 Cytochrome P-450 CYP2C9 Proteins 0.000 description 1
- 102100029363 Cytochrome P450 2C19 Human genes 0.000 description 1
- RGHNJXZEOKUKBD-UHFFFAOYSA-N D-gluconic acid Natural products OCC(O)C(O)C(O)C(O)C(O)=O RGHNJXZEOKUKBD-UHFFFAOYSA-N 0.000 description 1
- XBPCUCUWBYBCDP-UHFFFAOYSA-N Dicyclohexylamine Chemical compound C1CCCCC1NC1CCCCC1 XBPCUCUWBYBCDP-UHFFFAOYSA-N 0.000 description 1
- ZAFNJMIOTHYJRJ-UHFFFAOYSA-N Diisopropyl ether Chemical compound CC(C)OC(C)C ZAFNJMIOTHYJRJ-UHFFFAOYSA-N 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 208000001382 Experimental Melanoma Diseases 0.000 description 1
- 102000009109 Fc receptors Human genes 0.000 description 1
- 108010087819 Fc receptors Proteins 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 229910004373 HOAc Inorganic materials 0.000 description 1
- 206010019695 Hepatic neoplasm Diseases 0.000 description 1
- 206010019799 Hepatitis viral Diseases 0.000 description 1
- 102000018713 Histocompatibility Antigens Class II Human genes 0.000 description 1
- 108010027412 Histocompatibility Antigens Class II Proteins 0.000 description 1
- 101100005713 Homo sapiens CD4 gene Proteins 0.000 description 1
- 101001012157 Homo sapiens Receptor tyrosine-protein kinase erbB-2 Proteins 0.000 description 1
- 101000738771 Homo sapiens Receptor-type tyrosine-protein phosphatase C Proteins 0.000 description 1
- 101000831496 Homo sapiens Toll-like receptor 3 Proteins 0.000 description 1
- 101000800483 Homo sapiens Toll-like receptor 8 Proteins 0.000 description 1
- 101000997832 Homo sapiens Tyrosine-protein kinase JAK2 Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 description 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- LPHGQDQBBGAPDZ-UHFFFAOYSA-N Isocaffeine Natural products CN1C(=O)N(C)C(=O)C2=C1N(C)C=N2 LPHGQDQBBGAPDZ-UHFFFAOYSA-N 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- HNDVDQJCIGZPNO-YFKPBYRVSA-N L-histidine Chemical compound OC(=O)[C@@H](N)CC1=CN=CN1 HNDVDQJCIGZPNO-YFKPBYRVSA-N 0.000 description 1
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 101710128836 Large T antigen Proteins 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 206010027465 Metastases to skin Diseases 0.000 description 1
- 102000004232 Mitogen-Activated Protein Kinase Kinases Human genes 0.000 description 1
- 108090000744 Mitogen-Activated Protein Kinase Kinases Proteins 0.000 description 1
- 101000574441 Mus musculus Alkaline phosphatase, germ cell type Proteins 0.000 description 1
- IQAPYWHMSWELBX-UHFFFAOYSA-N N-[2-[acetyl(methyl)amino]ethyl]-N-methylcarbamoyl chloride Chemical compound C(C)(=O)N(CCN(C(=O)Cl)C)C IQAPYWHMSWELBX-UHFFFAOYSA-N 0.000 description 1
- HTLZVHNRZJPSMI-UHFFFAOYSA-N N-ethylpiperidine Chemical compound CCN1CCCCC1 HTLZVHNRZJPSMI-UHFFFAOYSA-N 0.000 description 1
- MBBZMMPHUWSWHV-BDVNFPICSA-N N-methylglucamine Chemical compound CNC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO MBBZMMPHUWSWHV-BDVNFPICSA-N 0.000 description 1
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 1
- 239000006057 Non-nutritive feed additive Substances 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- YGYAWVDWMABLBF-UHFFFAOYSA-N Phosgene Chemical compound ClC(Cl)=O YGYAWVDWMABLBF-UHFFFAOYSA-N 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 239000004743 Polypropylene Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 101710089372 Programmed cell death protein 1 Proteins 0.000 description 1
- OFOBLEOULBTSOW-UHFFFAOYSA-N Propanedioic acid Natural products OC(=O)CC(O)=O OFOBLEOULBTSOW-UHFFFAOYSA-N 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 102100030086 Receptor tyrosine-protein kinase erbB-2 Human genes 0.000 description 1
- 102100037422 Receptor-type tyrosine-protein phosphatase C Human genes 0.000 description 1
- 206010070308 Refractory cancer Diseases 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 102000003714 TNF receptor-associated factor 6 Human genes 0.000 description 1
- 108090000009 TNF receptor-associated factor 6 Proteins 0.000 description 1
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 1
- JLRGJRBPOGGCBT-UHFFFAOYSA-N Tolbutamide Chemical compound CCCCNC(=O)NS(=O)(=O)C1=CC=C(C)C=C1 JLRGJRBPOGGCBT-UHFFFAOYSA-N 0.000 description 1
- 108010060818 Toll-Like Receptor 9 Proteins 0.000 description 1
- 102000008235 Toll-Like Receptor 9 Human genes 0.000 description 1
- 102100024324 Toll-like receptor 3 Human genes 0.000 description 1
- 102100033110 Toll-like receptor 8 Human genes 0.000 description 1
- 108091005906 Type I transmembrane proteins Proteins 0.000 description 1
- 102100033444 Tyrosine-protein kinase JAK2 Human genes 0.000 description 1
- 102000005789 Vascular Endothelial Growth Factors Human genes 0.000 description 1
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 208000033559 Waldenström macroglobulinemia Diseases 0.000 description 1
- LJOOWESTVASNOG-UFJKPHDISA-N [(1s,3r,4ar,7s,8s,8as)-3-hydroxy-8-[2-[(4r)-4-hydroxy-6-oxooxan-2-yl]ethyl]-7-methyl-1,2,3,4,4a,7,8,8a-octahydronaphthalen-1-yl] (2s)-2-methylbutanoate Chemical compound C([C@H]1[C@@H](C)C=C[C@H]2C[C@@H](O)C[C@@H]([C@H]12)OC(=O)[C@@H](C)CC)CC1C[C@@H](O)CC(=O)O1 LJOOWESTVASNOG-UFJKPHDISA-N 0.000 description 1
- SORGEQQSQGNZFI-UHFFFAOYSA-N [azido(phenoxy)phosphoryl]oxybenzene Chemical compound C=1C=CC=CC=1OP(=O)(N=[N+]=[N-])OC1=CC=CC=C1 SORGEQQSQGNZFI-UHFFFAOYSA-N 0.000 description 1
- 210000003489 abdominal muscle Anatomy 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 235000011054 acetic acid Nutrition 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 102000035181 adaptor proteins Human genes 0.000 description 1
- 108091005764 adaptor proteins Proteins 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 210000002534 adenoid Anatomy 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 150000001350 alkyl halides Chemical class 0.000 description 1
- 230000029936 alkylation Effects 0.000 description 1
- 238000005804 alkylation reaction Methods 0.000 description 1
- 201000009961 allergic asthma Diseases 0.000 description 1
- 208000026935 allergic disease Diseases 0.000 description 1
- 230000007815 allergy Effects 0.000 description 1
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 239000002269 analeptic agent Substances 0.000 description 1
- 230000002491 angiogenic effect Effects 0.000 description 1
- 229960004977 anhydrous lactose Drugs 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 230000000840 anti-viral effect Effects 0.000 description 1
- 210000000612 antigen-presenting cell Anatomy 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 235000006708 antioxidants Nutrition 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 229960003121 arginine Drugs 0.000 description 1
- 239000012300 argon atmosphere Substances 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 125000003725 azepanyl group Chemical group 0.000 description 1
- HGQULGDOROIPJN-UHFFFAOYSA-N azetidin-1-ium;chloride Chemical compound Cl.C1CNC1 HGQULGDOROIPJN-UHFFFAOYSA-N 0.000 description 1
- 125000004069 aziridinyl group Chemical group 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 229940064804 betadine Drugs 0.000 description 1
- 229960003237 betaine Drugs 0.000 description 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 239000003809 bile pigment Substances 0.000 description 1
- 239000013060 biological fluid Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 125000001246 bromo group Chemical group Br* 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000004067 bulking agent Substances 0.000 description 1
- KDYFGRWQOYBRFD-NUQCWPJISA-N butanedioic acid Chemical compound O[14C](=O)CC[14C](O)=O KDYFGRWQOYBRFD-NUQCWPJISA-N 0.000 description 1
- YAUWHXKOYHADTR-UHFFFAOYSA-N butyl n-methylcarbamate Chemical compound CCCCOC(=O)NC YAUWHXKOYHADTR-UHFFFAOYSA-N 0.000 description 1
- 229910000024 caesium carbonate Inorganic materials 0.000 description 1
- 229960001948 caffeine Drugs 0.000 description 1
- VJEONQKOZGKCAK-UHFFFAOYSA-N caffeine Natural products CN1C(=O)N(C)C(=O)C2=C1C=CN2C VJEONQKOZGKCAK-UHFFFAOYSA-N 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 125000001951 carbamoylamino group Chemical group C(N)(=O)N* 0.000 description 1
- 125000004432 carbon atom Chemical group C* 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000010382 chemical cross-linking Methods 0.000 description 1
- 230000010109 chemoembolization Effects 0.000 description 1
- 125000001309 chloro group Chemical group Cl* 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 235000013985 cinnamic acid Nutrition 0.000 description 1
- 229930016911 cinnamic acid Natural products 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 229940127204 compound 29 Drugs 0.000 description 1
- 238000013170 computed tomography imaging Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 235000010947 crosslinked sodium carboxy methyl cellulose Nutrition 0.000 description 1
- 238000002681 cryosurgery Methods 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 210000005220 cytoplasmic tail Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000001934 delay Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000010511 deprotection reaction Methods 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 125000005959 diazepanyl group Chemical group 0.000 description 1
- PQZTVWVYCLIIJY-UHFFFAOYSA-N diethyl(propyl)amine Chemical compound CCCN(CC)CC PQZTVWVYCLIIJY-UHFFFAOYSA-N 0.000 description 1
- 125000004852 dihydrofuranyl group Chemical group O1C(CC=C1)* 0.000 description 1
- 125000005050 dihydrooxazolyl group Chemical group O1C(NC=C1)* 0.000 description 1
- 125000005043 dihydropyranyl group Chemical group O1C(CCC=C1)* 0.000 description 1
- MKRTXPORKIRPDG-UHFFFAOYSA-N diphenylphosphoryl azide Chemical compound C=1C=CC=CC=1P(=O)(N=[N+]=[N-])C1=CC=CC=C1 MKRTXPORKIRPDG-UHFFFAOYSA-N 0.000 description 1
- YWEUIGNSBFLMFL-UHFFFAOYSA-N diphosphonate Chemical compound O=P(=O)OP(=O)=O YWEUIGNSBFLMFL-UHFFFAOYSA-N 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 230000009429 distress Effects 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000009510 drug design Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000001163 endosome Anatomy 0.000 description 1
- 230000006862 enzymatic digestion Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 230000008029 eradication Effects 0.000 description 1
- CCIVGXIOQKPBKL-UHFFFAOYSA-M ethanesulfonate Chemical compound CCS([O-])(=O)=O CCIVGXIOQKPBKL-UHFFFAOYSA-M 0.000 description 1
- 125000001301 ethoxy group Chemical group [H]C([H])([H])C([H])([H])O* 0.000 description 1
- CISYAICWRKYWSD-ZETCQYMHSA-N ethyl (2S)-2-[carbonochloridoyl(methyl)amino]-3-methylbutanoate Chemical compound ClC(=O)N([C@H](C(=O)OCC)C(C)C)C CISYAICWRKYWSD-ZETCQYMHSA-N 0.000 description 1
- HJDHAECPVQKTGY-NSHDSACASA-N ethyl (2S)-2-[carbonochloridoyl(methyl)amino]-3-phenylpropanoate Chemical compound ClC(=O)N([C@H](C(=O)OCC)CC1=CC=CC=C1)C HJDHAECPVQKTGY-NSHDSACASA-N 0.000 description 1
- MJGADRCIWZVGOF-QMMMGPOBSA-N ethyl (2S)-2-[carbonochloridoyl(methyl)amino]-4-methylpentanoate Chemical compound ClC(=O)N([C@H](C(=O)OCC)CC(C)C)C MJGADRCIWZVGOF-QMMMGPOBSA-N 0.000 description 1
- MJTIXJCZZQPZMF-YFKPBYRVSA-N ethyl (2S)-2-[carbonochloridoyl(methyl)amino]propanoate Chemical compound ClC(=O)N([C@H](C(=O)OCC)C)C MJTIXJCZZQPZMF-YFKPBYRVSA-N 0.000 description 1
- QKRGDUOBYWVMFC-UHFFFAOYSA-N ethyl 3-[carbonochloridoyl(methyl)amino]propanoate Chemical compound ClC(=O)N(CCC(=O)OCC)C QKRGDUOBYWVMFC-UHFFFAOYSA-N 0.000 description 1
- RIFGWPKJUGCATF-UHFFFAOYSA-N ethyl chloroformate Chemical compound CCOC(Cl)=O RIFGWPKJUGCATF-UHFFFAOYSA-N 0.000 description 1
- 229940012017 ethylenediamine Drugs 0.000 description 1
- 239000012091 fetal bovine serum Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 125000001153 fluoro group Chemical group F* 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 239000001530 fumaric acid Substances 0.000 description 1
- 235000011087 fumaric acid Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000000174 gluconic acid Substances 0.000 description 1
- 235000012208 gluconic acid Nutrition 0.000 description 1
- 229960002442 glucosamine Drugs 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- 244000144993 groups of animals Species 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 230000007773 growth pattern Effects 0.000 description 1
- 210000002989 hepatic vein Anatomy 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 229960002885 histidine Drugs 0.000 description 1
- XGIHQYAWBCFNPY-AZOCGYLKSA-N hydrabamine Chemical compound C([C@@H]12)CC3=CC(C(C)C)=CC=C3[C@@]2(C)CCC[C@@]1(C)CNCCNC[C@@]1(C)[C@@H]2CCC3=CC(C(C)C)=CC=C3[C@@]2(C)CCC1 XGIHQYAWBCFNPY-AZOCGYLKSA-N 0.000 description 1
- MHAJPDPJQMAIIY-UHFFFAOYSA-N hydrogen peroxide Substances OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 1
- 125000002632 imidazolidinyl group Chemical group 0.000 description 1
- 125000002636 imidazolinyl group Chemical group 0.000 description 1
- DOUYETYNHWVLEO-UHFFFAOYSA-N imiquimod Chemical compound C1=CC=CC2=C3N(CC(C)C)C=NC3=C(N)N=C21 DOUYETYNHWVLEO-UHFFFAOYSA-N 0.000 description 1
- 229960002751 imiquimod Drugs 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 230000031261 interleukin-10 production Effects 0.000 description 1
- 230000004073 interleukin-2 production Effects 0.000 description 1
- 238000001361 intraarterial administration Methods 0.000 description 1
- 102000027411 intracellular receptors Human genes 0.000 description 1
- 108091008582 intracellular receptors Proteins 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 125000002346 iodo group Chemical group I* 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 210000004153 islets of langerhan Anatomy 0.000 description 1
- 239000012948 isocyanate Substances 0.000 description 1
- 150000002513 isocyanates Chemical class 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 125000003253 isopropoxy group Chemical group [H]C([H])([H])C([H])(O*)C([H])([H])[H] 0.000 description 1
- JJWLVOIRVHMVIS-UHFFFAOYSA-N isopropylamine Chemical compound CC(C)N JJWLVOIRVHMVIS-UHFFFAOYSA-N 0.000 description 1
- 125000003965 isoxazolidinyl group Chemical group 0.000 description 1
- 210000004731 jugular vein Anatomy 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 229960001632 labetalol Drugs 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 238000002514 liquid chromatography mass spectrum Methods 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 239000012931 lyophilized formulation Substances 0.000 description 1
- 229960003646 lysine Drugs 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- VZCYOOQTPOCHFL-UPHRSURJSA-N maleic acid Chemical compound OC(=O)\C=C/C(O)=O VZCYOOQTPOCHFL-UPHRSURJSA-N 0.000 description 1
- 239000011976 maleic acid Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 210000001758 mesenteric vein Anatomy 0.000 description 1
- 229940098779 methanesulfonic acid Drugs 0.000 description 1
- WBYWAXJHAXSJNI-UHFFFAOYSA-N methyl p-hydroxycinnamate Natural products OC(=O)C=CC1=CC=CC=C1 WBYWAXJHAXSJNI-UHFFFAOYSA-N 0.000 description 1
- NQMRYBIKMRVZLB-UHFFFAOYSA-N methylamine hydrochloride Chemical compound [Cl-].[NH3+]C NQMRYBIKMRVZLB-UHFFFAOYSA-N 0.000 description 1
- 230000004089 microcirculation Effects 0.000 description 1
- 230000003228 microsomal effect Effects 0.000 description 1
- 150000007522 mineralic acids Chemical class 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 125000002950 monocyclic group Chemical group 0.000 description 1
- 125000002757 morpholinyl group Chemical group 0.000 description 1
- APRJFNLVTJWEPP-UHFFFAOYSA-M n,n-diethylcarbamate Chemical compound CCN(CC)C([O-])=O APRJFNLVTJWEPP-UHFFFAOYSA-M 0.000 description 1
- UDZCEFCJEGGQOJ-UHFFFAOYSA-N n-(2-methoxyethyl)propan-1-amine Chemical compound CCCNCCOC UDZCEFCJEGGQOJ-UHFFFAOYSA-N 0.000 description 1
- 125000004108 n-butyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- VGEMYWDUTPQWBN-UHFFFAOYSA-N n-ethyl-2-methoxyethanamine Chemical compound CCNCCOC VGEMYWDUTPQWBN-UHFFFAOYSA-N 0.000 description 1
- QHCCDDQKNUYGNC-UHFFFAOYSA-N n-ethylbutan-1-amine Chemical compound CCCCNCC QHCCDDQKNUYGNC-UHFFFAOYSA-N 0.000 description 1
- XCVNDBIXFPGMIW-UHFFFAOYSA-N n-ethylpropan-1-amine Chemical compound CCCNCC XCVNDBIXFPGMIW-UHFFFAOYSA-N 0.000 description 1
- GRRYSIXDUIAUGY-UHFFFAOYSA-N n-methylcarbamoyl chloride Chemical compound CNC(Cl)=O GRRYSIXDUIAUGY-UHFFFAOYSA-N 0.000 description 1
- XHFGWHUWQXTGAT-UHFFFAOYSA-N n-methylpropan-2-amine Chemical compound CNC(C)C XHFGWHUWQXTGAT-UHFFFAOYSA-N 0.000 description 1
- 210000000581 natural killer T-cell Anatomy 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 230000000683 nonmetastatic effect Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 125000002347 octyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 239000006186 oral dosage form Substances 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 125000005961 oxazepanyl group Chemical group 0.000 description 1
- 125000000160 oxazolidinyl group Chemical group 0.000 description 1
- 125000003566 oxetanyl group Chemical group 0.000 description 1
- 125000000466 oxiranyl group Chemical group 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- WLJNZVDCPSBLRP-UHFFFAOYSA-N pamoic acid Chemical compound C1=CC=C2C(CC=3C4=CC=CC=C4C=C(C=3O)C(=O)O)=C(O)C(C(O)=O)=CC2=C1 WLJNZVDCPSBLRP-UHFFFAOYSA-N 0.000 description 1
- 238000012753 partial hepatectomy Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 210000004976 peripheral blood cell Anatomy 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 229940124531 pharmaceutical excipient Drugs 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 125000000951 phenoxy group Chemical group [H]C1=C([H])C([H])=C(O*)C([H])=C1[H] 0.000 description 1
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 1
- DLYUQMMRRRQYAE-UHFFFAOYSA-N phosphorus pentoxide Inorganic materials O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 229920000768 polyamine Polymers 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000137 polyphosphoric acid Polymers 0.000 description 1
- 229920001155 polypropylene Polymers 0.000 description 1
- 230000001124 posttranscriptional effect Effects 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 235000015320 potassium carbonate Nutrition 0.000 description 1
- 235000011181 potassium carbonates Nutrition 0.000 description 1
- 239000008057 potassium phosphate buffer Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 1
- 229960004919 procaine Drugs 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- AQDNAHPBMCJXJD-LBPRGKRZSA-N propan-2-yl (2S)-2-[carbonochloridoyl(methyl)amino]-3-phenylpropanoate Chemical compound ClC(=O)N([C@H](C(=O)OC(C)C)CC1=CC=CC=C1)C AQDNAHPBMCJXJD-LBPRGKRZSA-N 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- LVTJOONKWUXEFR-FZRMHRINSA-N protoneodioscin Natural products O(C[C@@H](CC[C@]1(O)[C@H](C)[C@@H]2[C@]3(C)[C@H]([C@H]4[C@@H]([C@]5(C)C(=CC4)C[C@@H](O[C@@H]4[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@@H](O)[C@H](O[C@H]6[C@@H](O)[C@@H](O)[C@@H](O)[C@H](C)O6)[C@H](CO)O4)CC5)CC3)C[C@@H]2O1)C)[C@H]1[C@H](O)[C@H](O)[C@H](O)[C@@H](CO)O1 LVTJOONKWUXEFR-FZRMHRINSA-N 0.000 description 1
- 150000003212 purines Chemical class 0.000 description 1
- 125000003072 pyrazolidinyl group Chemical group 0.000 description 1
- NYCVCXMSZNOGDH-UHFFFAOYSA-N pyrrolidine-1-carboxylic acid Chemical compound OC(=O)N1CCCC1 NYCVCXMSZNOGDH-UHFFFAOYSA-N 0.000 description 1
- 229940107700 pyruvic acid Drugs 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 125000004621 quinuclidinyl group Chemical group N12C(CC(CC1)CC2)* 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 208000016691 refractory malignant neoplasm Diseases 0.000 description 1
- ZOPOQLDXFHBOIH-UHFFFAOYSA-N regorafenib hydrate Chemical compound O.C1=NC(C(=O)NC)=CC(OC=2C=C(F)C(NC(=O)NC=3C=C(C(Cl)=CC=3)C(F)(F)F)=CC=2)=C1 ZOPOQLDXFHBOIH-UHFFFAOYSA-N 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 1
- 150000003335 secondary amines Chemical class 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 238000010898 silica gel chromatography Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 229910000104 sodium hydride Inorganic materials 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 210000001179 synovial fluid Anatomy 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
- 235000002906 tartaric acid Nutrition 0.000 description 1
- 125000004213 tert-butoxy group Chemical group [H]C([H])([H])C(O*)(C([H])([H])[H])C([H])([H])[H] 0.000 description 1
- ASXJGFBXIYFSJC-UHFFFAOYSA-N tert-butyl n-[2-[carbonochloridoyl(methyl)amino]ethyl]-n-methylcarbamate Chemical compound ClC(=O)N(C)CCN(C)C(=O)OC(C)(C)C ASXJGFBXIYFSJC-UHFFFAOYSA-N 0.000 description 1
- ISXSCDLOGDJUNJ-UHFFFAOYSA-N tert-butyl prop-2-enoate Chemical compound CC(C)(C)OC(=O)C=C ISXSCDLOGDJUNJ-UHFFFAOYSA-N 0.000 description 1
- 150000003512 tertiary amines Chemical class 0.000 description 1
- 125000003718 tetrahydrofuranyl group Chemical group 0.000 description 1
- 125000001412 tetrahydropyranyl group Chemical group 0.000 description 1
- 125000004853 tetrahydropyridinyl group Chemical group N1(CCCC=C1)* 0.000 description 1
- 125000005958 tetrahydrothienyl group Chemical group 0.000 description 1
- 125000004632 tetrahydrothiopyranyl group Chemical group S1C(CCCC1)* 0.000 description 1
- 229960004559 theobromine Drugs 0.000 description 1
- 125000001984 thiazolidinyl group Chemical group 0.000 description 1
- 125000004568 thiomorpholinyl group Chemical group 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 229960005371 tolbutamide Drugs 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- YFTHZRPMJXBUME-UHFFFAOYSA-N tripropylamine Chemical compound CCCN(CCC)CCC YFTHZRPMJXBUME-UHFFFAOYSA-N 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000006433 tumor necrosis factor production Effects 0.000 description 1
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- AQLJVWUFPCUVLO-UHFFFAOYSA-N urea hydrogen peroxide Chemical compound OO.NC(N)=O AQLJVWUFPCUVLO-UHFFFAOYSA-N 0.000 description 1
- 230000002792 vascular Effects 0.000 description 1
- 201000001862 viral hepatitis Diseases 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
- A61K31/52—Purines, e.g. adenine
- A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D473/00—Heterocyclic compounds containing purine ring systems
- C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
- C07D473/24—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 one nitrogen and one sulfur atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
- C07K16/2818—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
Abstract
The present invention relates to compounds of formula (I), wherein R1, R2 and R3 are as described herein, and their prodrugs or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for (use in) the treatment and/or prophylaxis of liver cancer.
Description
2 FOR THE TREATMENT AND PROPHYLAXIS OF LIVER CANCER
The present invention relates to novel sulfonimidoylpurinones derivatives that have in vivo Toll-like receptor agonism activity, for (use in) the treatment and/or prophylaxis of liver cancer BACKGROUND
Liver cancer is the fifth most common form of cancer. Each year, approximately 750,000 cases are diagnosed and about 700,000 people die from the disease each year, making it the third most common cause of cancer death in the world (Ferlay et al., Int. J.
Cancer 127:2893-2917 (2010)). In the United States, the incidence of primary liver cancer has been rising, and while some progress has been made in detecting and treating localized disease, the five year survival rate for late stage liver cancer is still well below 10%
(American-Cancer- Society. 2012. Cancer Facts & Figures 2012. Atlanta:
American Cancer Society).
Established treatments for liver cancer include surgical removal of the part of the liver containing the tumor (partial hepatectomy), liver transplantation, transcatheter arterial chemoembolization (TACE), in situ tumor destruction by various methods such as radiofrequency ablation (RFA) or cryosurgery and administration of Sorafenib.
Treatment options for late stage liver patients are limited. Thus, effective treatments of liver cancer remain a significant unmet medical need.
The present invention relates to compounds of formula (I), N H 2 .....-R3 N N
----- > 0 x N
0=S N
1 1 ' 2 R
R (I), wherein Rl to R3 are described below, or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
Toll-like receptors (TLRs) detect a wide range of conserved pathogen-associated molecular patterns (PAMPs). They play an important role of sensing invading pathogens and subsequent initiation of innate immune responses. There are 10 known members of the TLR family in human, which are type I transmembrane proteins featuring an extracellular leucine-rich domain and a cytoplasmic tail that contains a conserved Toll/
interleukin (IL)-1 receptor (TIR) domain. Within this family, TLR3, TLR7, TLR8 and TLR9 are located within endosomes.
TLR7 can be activated by binding to a specific small molecule ligand (i.e., agonist) or its native ligand (i.e., single-stranded RNA, ssRNA). Following binding of ssRNA to TLR7, the receptor in its dimerized form is believed to undergo a structural change leading to the subsequent recruitment of adapter proteins at its cytoplasmic domain, including the myeloid differentiation primary response gene 88 (MyD88).
Following the initiation of the receptor signalling cascade via the MyD88 pathway, cytoplasmic transcription factors such as interferon regulatory factor 7 (IRF-7) and nuclear factor kappa B (NF-KB) are activated. These transcription factors then translocate to the nucleus and initiate the transcription of various genes, e.g., IFN-a and other antiviral cytokine genes.
W0201772662 relates to TLR7 agonist-anti HER2 conjugates for the treatment of positive cancers. Hotz et al, Oncoimmunology 2012, 227-228 relates to cancer treatment with TLR7 agonists. However so far no TLR7 agonist is used systemically for the treatment of cancer. Only topical TLR7 agonist imiquimod is known to induce immune-mediated rejection of skin metastases in patients with breast cancer (Adams S., Kozhaya L., Martiniuk F., Meng T.C., Chiriboga L., Liebes L., Hochman T., Shuman N., Axelrod D., Speyer J., et al. Clin. Cancer Res. 2012;18:6748-6757.
SUMMARY OF THE INVENTION
The present invention relates to a series of novel 6-amino-2-sulfonimidoy1-9-substituted-7-substituted-purin-8-one compounds with Toll-like receptor agonistic activity and their prodrugs for use in the treatment or prophylaxis (prevention) of liver cancer.
It was found out that the potent and safe TLR7 agonist prodrugs described herein are effective in the treatment of liver cancer either alone or in combination with other agents.
The present invention provides a series of novel 6-amino-2-sulfonimidoy1-9-substituted-7-substituted-purin-8-one compounds that have Toll-like receptor agonistic activity and their prodrugs. The invention also provides the bio-activity of such compounds
The present invention relates to novel sulfonimidoylpurinones derivatives that have in vivo Toll-like receptor agonism activity, for (use in) the treatment and/or prophylaxis of liver cancer BACKGROUND
Liver cancer is the fifth most common form of cancer. Each year, approximately 750,000 cases are diagnosed and about 700,000 people die from the disease each year, making it the third most common cause of cancer death in the world (Ferlay et al., Int. J.
Cancer 127:2893-2917 (2010)). In the United States, the incidence of primary liver cancer has been rising, and while some progress has been made in detecting and treating localized disease, the five year survival rate for late stage liver cancer is still well below 10%
(American-Cancer- Society. 2012. Cancer Facts & Figures 2012. Atlanta:
American Cancer Society).
Established treatments for liver cancer include surgical removal of the part of the liver containing the tumor (partial hepatectomy), liver transplantation, transcatheter arterial chemoembolization (TACE), in situ tumor destruction by various methods such as radiofrequency ablation (RFA) or cryosurgery and administration of Sorafenib.
Treatment options for late stage liver patients are limited. Thus, effective treatments of liver cancer remain a significant unmet medical need.
The present invention relates to compounds of formula (I), N H 2 .....-R3 N N
----- > 0 x N
0=S N
1 1 ' 2 R
R (I), wherein Rl to R3 are described below, or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
Toll-like receptors (TLRs) detect a wide range of conserved pathogen-associated molecular patterns (PAMPs). They play an important role of sensing invading pathogens and subsequent initiation of innate immune responses. There are 10 known members of the TLR family in human, which are type I transmembrane proteins featuring an extracellular leucine-rich domain and a cytoplasmic tail that contains a conserved Toll/
interleukin (IL)-1 receptor (TIR) domain. Within this family, TLR3, TLR7, TLR8 and TLR9 are located within endosomes.
TLR7 can be activated by binding to a specific small molecule ligand (i.e., agonist) or its native ligand (i.e., single-stranded RNA, ssRNA). Following binding of ssRNA to TLR7, the receptor in its dimerized form is believed to undergo a structural change leading to the subsequent recruitment of adapter proteins at its cytoplasmic domain, including the myeloid differentiation primary response gene 88 (MyD88).
Following the initiation of the receptor signalling cascade via the MyD88 pathway, cytoplasmic transcription factors such as interferon regulatory factor 7 (IRF-7) and nuclear factor kappa B (NF-KB) are activated. These transcription factors then translocate to the nucleus and initiate the transcription of various genes, e.g., IFN-a and other antiviral cytokine genes.
W0201772662 relates to TLR7 agonist-anti HER2 conjugates for the treatment of positive cancers. Hotz et al, Oncoimmunology 2012, 227-228 relates to cancer treatment with TLR7 agonists. However so far no TLR7 agonist is used systemically for the treatment of cancer. Only topical TLR7 agonist imiquimod is known to induce immune-mediated rejection of skin metastases in patients with breast cancer (Adams S., Kozhaya L., Martiniuk F., Meng T.C., Chiriboga L., Liebes L., Hochman T., Shuman N., Axelrod D., Speyer J., et al. Clin. Cancer Res. 2012;18:6748-6757.
SUMMARY OF THE INVENTION
The present invention relates to a series of novel 6-amino-2-sulfonimidoy1-9-substituted-7-substituted-purin-8-one compounds with Toll-like receptor agonistic activity and their prodrugs for use in the treatment or prophylaxis (prevention) of liver cancer.
It was found out that the potent and safe TLR7 agonist prodrugs described herein are effective in the treatment of liver cancer either alone or in combination with other agents.
The present invention provides a series of novel 6-amino-2-sulfonimidoy1-9-substituted-7-substituted-purin-8-one compounds that have Toll-like receptor agonistic activity and their prodrugs. The invention also provides the bio-activity of such compounds
- 3 -to induce cytokine/chemokine release, SEAP level increase by activating Toll-like receptors, such as TLR7 receptor, the metabolic conversion of prodrugs to parent compounds in the presence of human hepatocytes, and the therapeutic or prophylactic use of such compounds and their pharmaceutical compositions comprising these compounds and their prodrugs to treat or prevent liver cancer. The present invention also provides compounds with superior activity. In addition, the compounds of formula (I) also show good solubility and PK profiles.
The present invention relates to novel compounds of formula (I), N H 2 ....-R3 N---N> 0 II..õ...s....... ...õ-------N
0=S N
1 1 \ 2 R
R (I), wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
The present invention relates to novel compounds of formula (I), N H 2 ....-R3 N---N> 0 II..õ...s....... ...õ-------N
0=S N
1 1 \ 2 R
R (I), wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
- 4 -6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
These prodrug compounds are especially useful for the treatment of liver cancer as they are activated (converted into their active form) in the liver. They show valuable anti-tumor efficacy in vivo in liver cancer cell models (either alone or in combination with anti-PD1/PD1 antibodies or with anti-angiogenic agents) and in vitro against liver cancer cells (by activation of peripheral blood cells and/or factors).
The invention also relates to their use for the manufacture of a medicament for the treatment or prophylaxis of liver cancer, medicaments based on a compound in accordance with the invention for the treatment or prophylaxis of liver cancer.
Accordingly, the compounds of formula (I) are useful for the treatment or prophylaxis of liver cancer, especially for the treatment or prophylaxis of hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer.
DETAILED DESCRIPTION OF THE INVENTION
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Furthermore, the following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the invention.
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
These prodrug compounds are especially useful for the treatment of liver cancer as they are activated (converted into their active form) in the liver. They show valuable anti-tumor efficacy in vivo in liver cancer cell models (either alone or in combination with anti-PD1/PD1 antibodies or with anti-angiogenic agents) and in vitro against liver cancer cells (by activation of peripheral blood cells and/or factors).
The invention also relates to their use for the manufacture of a medicament for the treatment or prophylaxis of liver cancer, medicaments based on a compound in accordance with the invention for the treatment or prophylaxis of liver cancer.
Accordingly, the compounds of formula (I) are useful for the treatment or prophylaxis of liver cancer, especially for the treatment or prophylaxis of hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer.
DETAILED DESCRIPTION OF THE INVENTION
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Furthermore, the following definitions are set forth to illustrate and define the meaning and scope of the various terms used to describe the invention.
- 5 -The term "Ci_6alkyl" denotes a saturated, linear or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl and the like. Particular "C1_6alkyl"
groups are methyl, ethyl and n-propyl.
The term "Ci_6alkoxy" denotes a group of the formula Ci_6alky1-0-. Examples of C1_ 6a1k0xy group include, but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy. Particular "Ci_6alkoxy" groups are methoxy, ethoxy and isopropoxy. A more particular Ci_6alkoxy group is ethoxy.
The term "halogen" and "halo" are used interchangeably herein and denote fluoro, chloro, bromo, or iodo.
The term "heterocyclyl" denotes a monovalent saturated or partly unsaturated mono or bicyclic ring system of 3 to 10 ring atoms, comprising 1 to 5 ring heteroatoms selected from N, 0 and S, the remaining ring atoms being carbon. In particular embodiments, heterocyclyl is a monovalent saturated monocyclic ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, 0 and S, the remaining ring atoms being carbon. Examples for monocyclic saturated heterocyclyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, dimethylpyrrolidinyl, ethoxycarbonylpyrrolidinyl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, dioxothiomorpholinyl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Monocyclic saturated heterocyclyl can be further substituted by one to three substituents independently selected from halogen, Ci_6alkyl and C1_6alkoxycarbonyl. Examples for substituted monocyclic saturated heterocyclyl are 4-methylpiperazinyl, dimethylpyrrolidinyl, ethoxycarbonylpyrrolidinyl, difluoropyrrolidinyl, fluoro(methyl)pyrrolidinyl. Examples for bicyclic saturated heterocyclyl are azabicyclo[3.2.1]octyl, quinuclidinyl, oxaazabicyclo[3.2.1]octyl, azabicyclo[3.3.1]nonyl, oxaazabicyclo[3.3.1]nonyl, thiaazabicyclo[3.3.1]nonyl, azaspiro[3.3]heptanyl and oxaazaspiro[3.3]heptanyl. Examples for partly unsaturated heterocyclyl are dihydrofuryl, imidazolinyl, dihydrooxazolyl, tetrahydropyridinyl and dihydropyranyl.
The term "carbonyl" alone or in combination refers to the group -C(0)-.
groups are methyl, ethyl and n-propyl.
The term "Ci_6alkoxy" denotes a group of the formula Ci_6alky1-0-. Examples of C1_ 6a1k0xy group include, but not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy and tert-butoxy. Particular "Ci_6alkoxy" groups are methoxy, ethoxy and isopropoxy. A more particular Ci_6alkoxy group is ethoxy.
The term "halogen" and "halo" are used interchangeably herein and denote fluoro, chloro, bromo, or iodo.
The term "heterocyclyl" denotes a monovalent saturated or partly unsaturated mono or bicyclic ring system of 3 to 10 ring atoms, comprising 1 to 5 ring heteroatoms selected from N, 0 and S, the remaining ring atoms being carbon. In particular embodiments, heterocyclyl is a monovalent saturated monocyclic ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, 0 and S, the remaining ring atoms being carbon. Examples for monocyclic saturated heterocyclyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, dimethylpyrrolidinyl, ethoxycarbonylpyrrolidinyl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, dioxothiomorpholinyl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Monocyclic saturated heterocyclyl can be further substituted by one to three substituents independently selected from halogen, Ci_6alkyl and C1_6alkoxycarbonyl. Examples for substituted monocyclic saturated heterocyclyl are 4-methylpiperazinyl, dimethylpyrrolidinyl, ethoxycarbonylpyrrolidinyl, difluoropyrrolidinyl, fluoro(methyl)pyrrolidinyl. Examples for bicyclic saturated heterocyclyl are azabicyclo[3.2.1]octyl, quinuclidinyl, oxaazabicyclo[3.2.1]octyl, azabicyclo[3.3.1]nonyl, oxaazabicyclo[3.3.1]nonyl, thiaazabicyclo[3.3.1]nonyl, azaspiro[3.3]heptanyl and oxaazaspiro[3.3]heptanyl. Examples for partly unsaturated heterocyclyl are dihydrofuryl, imidazolinyl, dihydrooxazolyl, tetrahydropyridinyl and dihydropyranyl.
The term "carbonyl" alone or in combination refers to the group -C(0)-.
- 6 -The term "Ci_6alkylcarbonyl" refers to a group Ci_6alkyl-C(0)-, wherein the "Ci_ 6a1ky1" is as defined above. Particular "Ci_6alkylcarbonyl" group is acetyl.
The term "enantiomer" denotes two stereoisomers of a compound which are non-superimposable mirror images of one another.
The term "diastereomer" denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another.
Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
The term "pharmaceutically acceptable salts" denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts.
The term "pharmaceutically acceptable acid addition salt" denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicyclic acid.
The term "pharmaceutically acceptable base addition salt" denotes those pharmaceutically acceptable salts formed with an organic or inorganic base.
Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, and polyamine resins.
The term "enantiomer" denotes two stereoisomers of a compound which are non-superimposable mirror images of one another.
The term "diastereomer" denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another.
Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.
The term "pharmaceutically acceptable salts" denotes salts which are not biologically or otherwise undesirable. Pharmaceutically acceptable salts include both acid and base addition salts.
The term "pharmaceutically acceptable acid addition salt" denotes those pharmaceutically acceptable salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid, phosphoric acid, and organic acids selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, gluconic acid, lactic acid, pyruvic acid, oxalic acid, malic acid, maleic acid, maloneic acid, succinic acid, fumaric acid, tartaric acid, citric acid, aspartic acid, ascorbic acid, glutamic acid, anthranilic acid, benzoic acid, cinnamic acid, mandelic acid, embonic acid, phenylacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, and salicyclic acid.
The term "pharmaceutically acceptable base addition salt" denotes those pharmaceutically acceptable salts formed with an organic or inorganic base.
Examples of acceptable inorganic bases include sodium, potassium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts. Salts derived from pharmaceutically acceptable organic nontoxic bases includes salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperizine, piperidine, N-ethylpiperidine, and polyamine resins.
- 7 -Compounds of the general formula (I) and their prodrugs which contain one or several chiral centers can either be present as racemates, diastereomeric mixtures, or optically active single isomers. The racemates can be separated according to known methods into the enantiomers. Particularly, diastereomeric salts which can be separated by crystallization are formed from the racemic mixtures by reaction with an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphorsulfonic acid.
The term "prodrug" denotes a form or derivative of a compound which is metabolized in vivo, e.g., by biological fluids or enzymes by a subject after administration, into a pharmacologically active form of the compound in order to produce the desired pharmacological effect. Prodrugs are described e.g. in "The Organic Chemistry of Drug Design and Drug Action", by Richard B. Silverman, Academic Press, San Diego, 2004, Chapter 8 Prodrugs and Drug Delivery Systems, pp. 497-558.
"A pharmaceutically active metabolite" is intended to mean a pharmacologically active product produced through metabolism in the body of a specified compound or salt thereof. After entry into the body, most drugs are substrates for chemical reactions that may change their physical properties and biologic effects. These metabolic conversions, which usually affect the polarity of the compounds of the invention, alter the way in which drugs are distributed in and excreted from the body. However, in some cases, metabolism of a drug is required for therapeutic effect.
The term "therapeutically effective amount" denotes an amount of a compound or molecule of the present invention that, when administered to a subject, (i) treats or prevents the particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition or disorder described herein. The therapeutically effective amount will vary depending on the compound, the disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.
The term "pharmaceutical composition" denotes a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with
The term "prodrug" denotes a form or derivative of a compound which is metabolized in vivo, e.g., by biological fluids or enzymes by a subject after administration, into a pharmacologically active form of the compound in order to produce the desired pharmacological effect. Prodrugs are described e.g. in "The Organic Chemistry of Drug Design and Drug Action", by Richard B. Silverman, Academic Press, San Diego, 2004, Chapter 8 Prodrugs and Drug Delivery Systems, pp. 497-558.
"A pharmaceutically active metabolite" is intended to mean a pharmacologically active product produced through metabolism in the body of a specified compound or salt thereof. After entry into the body, most drugs are substrates for chemical reactions that may change their physical properties and biologic effects. These metabolic conversions, which usually affect the polarity of the compounds of the invention, alter the way in which drugs are distributed in and excreted from the body. However, in some cases, metabolism of a drug is required for therapeutic effect.
The term "therapeutically effective amount" denotes an amount of a compound or molecule of the present invention that, when administered to a subject, (i) treats or prevents the particular disease, condition or disorder, (ii) attenuates, ameliorates or eliminates one or more symptoms of the particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of the particular disease, condition or disorder described herein. The therapeutically effective amount will vary depending on the compound, the disease state being treated, the severity of the disease treated, the age and relative health of the subject, the route and form of administration, the judgement of the attending medical or veterinary practitioner, and other factors.
The term "pharmaceutical composition" denotes a mixture or solution comprising a therapeutically effective amount of an active pharmaceutical ingredient together with
- 8 -pharmaceutically acceptable excipients to be administered to a mammal, e.g., a human in need thereof.
The present invention relates to a compound of formula (I), N H 2 ....¨ R3 N --- N H N > 0 II ..õ...-sõ..... ...õ------.... N
0 = S N
1 1 \ 2 R
R (I), wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1- [6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
The present invention relates to a compound of formula (I), N H 2 ....¨ R3 N --- N H N > 0 II ..õ...-sõ..... ...õ------.... N
0 = S N
1 1 \ 2 R
R (I), wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1- [6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
- 9 -6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
A further embodiment of present invention is (ii) a compound of formula (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
pip erazinyl substituted by Ci_6alkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci_ 6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6a1ky1, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iii) a compound of formula (I), wherein Rl is ethyl or propyl;
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
A further embodiment of present invention is (ii) a compound of formula (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
pip erazinyl substituted by Ci_6alkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci_ 6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6a1ky1, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iii) a compound of formula (I), wherein Rl is ethyl or propyl;
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
- 10 -piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iii-1) a compound of formula (I), wherein Rl is ethyl or propyl;
R2 is benzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl,
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iii-1) a compound of formula (I), wherein Rl is ethyl or propyl;
R2 is benzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl,
- 11 -methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iv) a compound of formula (I), wherein R3 is azetidinyl, 4-methylpiperazinyl, piperidinylpiperidinyl, pyrrolidinyl, acetyl(methyl)aminoethyl(methyl)amino, bis(methoxyethyl)amino, butyl(ethyl)amino, butyl(methyl)amino, butyl(methyl)carbamoyloxyethyl(methyl)amino, diethylcarbamoyloxyethyl(methyl)amino, ethoxycarbonyl(methyl)aminoethyl(methyl)amino, ethoxycarbonylethyl(methyl)amino, ethoxycarbonylisobutyl(methyl)amino, ethoxycarbonylisopentyl(methyl)amino, ethoxycarbonylmethyl(methyl)amino, ethoxycarbonyloxyethyl(methyl)amino, ethoxycarbonyl(phenyl)ethyl(methyl)amino, ethyl(methyl)amino, isobutyl(methyl)amino, isopropoxycarbonylisopentyl(methyl)amino, isopropoxycarbonyl(phenyl)ethyl(methyl)amino, isopropyl(methyl)amino, methoxycarbonyl(methyl)aminoethyl(methyl)amino, methoxyethyl(ethyl)amino, methoxyethyl(methyl)amino, methoxyethyl(propyl)amino, methoxypropyl(methyl)amino, propyl(ethyl)amino, propyl(methyl)amino, propyl(methyl)carbamoyloxyethyl(methyl)amino, pyrrolidinylcarbamoyloxyethyl(methyl)amino, tert-butoxycarbonyl(methyl)aminoethyl(methyl)amino, tert-butoxycarbonylethyl(methyl)amino, tert-butoxycarbonylisopentyl(methyl)amino or tert-butoxycarbonyl(phenyl)ethyl(methyl)amino; or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (v) a compound of formula (I), wherein Rl is ethyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (vi) a compound of formula (I), wherein R2 is benzyl substituted by halogen or Ci_6alkyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (vii) a compound of formula (I), wherein R2 is bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl, for use in the treatment or prophylaxis of liver cancer.
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (iv) a compound of formula (I), wherein R3 is azetidinyl, 4-methylpiperazinyl, piperidinylpiperidinyl, pyrrolidinyl, acetyl(methyl)aminoethyl(methyl)amino, bis(methoxyethyl)amino, butyl(ethyl)amino, butyl(methyl)amino, butyl(methyl)carbamoyloxyethyl(methyl)amino, diethylcarbamoyloxyethyl(methyl)amino, ethoxycarbonyl(methyl)aminoethyl(methyl)amino, ethoxycarbonylethyl(methyl)amino, ethoxycarbonylisobutyl(methyl)amino, ethoxycarbonylisopentyl(methyl)amino, ethoxycarbonylmethyl(methyl)amino, ethoxycarbonyloxyethyl(methyl)amino, ethoxycarbonyl(phenyl)ethyl(methyl)amino, ethyl(methyl)amino, isobutyl(methyl)amino, isopropoxycarbonylisopentyl(methyl)amino, isopropoxycarbonyl(phenyl)ethyl(methyl)amino, isopropyl(methyl)amino, methoxycarbonyl(methyl)aminoethyl(methyl)amino, methoxyethyl(ethyl)amino, methoxyethyl(methyl)amino, methoxyethyl(propyl)amino, methoxypropyl(methyl)amino, propyl(ethyl)amino, propyl(methyl)amino, propyl(methyl)carbamoyloxyethyl(methyl)amino, pyrrolidinylcarbamoyloxyethyl(methyl)amino, tert-butoxycarbonyl(methyl)aminoethyl(methyl)amino, tert-butoxycarbonylethyl(methyl)amino, tert-butoxycarbonylisopentyl(methyl)amino or tert-butoxycarbonyl(phenyl)ethyl(methyl)amino; or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (v) a compound of formula (I), wherein Rl is ethyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (vi) a compound of formula (I), wherein R2 is benzyl substituted by halogen or Ci_6alkyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (vii) a compound of formula (I), wherein R2 is bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl, for use in the treatment or prophylaxis of liver cancer.
- 12 -A further embodiment of present invention is (vii-1) a compound of formula (I), wherein R2 is chlorobenzyl, fluorobenzyl or methylbenzyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (viii) a compound of formula (I), wherein R2 is bromobenzyl, chlorobenzyl or fluorobenzyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (viii-1) a compound of formula (I), wherein R2 is chlorobenzyl or fluorobenzyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (ix) a compound of formula (I), wherein R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is Ci_6alkyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (x) a compound of formula (I), wherein R3 is propyl(methyl)amino or ethyl(methyl)amino, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xi) a compound of formula (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being substituted by halogen or C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is C1_6alkyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xii) a compound of formula (I), wherein Rl is ethyl;
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xii-1) a compound of formula (I), wherein Rl is ethyl;
R2 is methylbenzyl, chlorobenzyl or fluorobenzyl;
A further embodiment of present invention is (viii) a compound of formula (I), wherein R2 is bromobenzyl, chlorobenzyl or fluorobenzyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (viii-1) a compound of formula (I), wherein R2 is chlorobenzyl or fluorobenzyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (ix) a compound of formula (I), wherein R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is Ci_6alkyl, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (x) a compound of formula (I), wherein R3 is propyl(methyl)amino or ethyl(methyl)amino, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xi) a compound of formula (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being substituted by halogen or C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is C1_6alkyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xii) a compound of formula (I), wherein Rl is ethyl;
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
A further embodiment of present invention is (xii-1) a compound of formula (I), wherein Rl is ethyl;
R2 is methylbenzyl, chlorobenzyl or fluorobenzyl;
- 13 -R3 is propyl(methyl)amino or ethyl(methyl)amino;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
Another embodiment of present invention is that (xiii) particular compounds of formula (I) are the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine- 1 -carb ony1)-2-(propylsulfonimidoyl)purin- 8 -one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
Another embodiment of present invention is that (xiii) particular compounds of formula (I) are the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine- 1 -carb ony1)-2-(propylsulfonimidoyl)purin- 8 -one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 14 -Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethyl]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethyl]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
- 15 -Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methyl-8-oxo-N-propy1-2[S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2- [S (R)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methyl-8-oxo-N-propy1-2[S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2- [S (R)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
- 16 -6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
- 17 -6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methyl- 8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
Another embodiment of present invention is that (xiv) more particular compounds of formula (I) are the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2- [S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methyl- 8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
Another embodiment of present invention is that (xiv) more particular compounds of formula (I) are the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2- [S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
- 18 -6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
- 19 -Another embodiment of present invention is that (xv) more particular compounds of formula (I) are the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
In some embodiments, compounds of present invention were tested and compared with the following reference compounds. As the most successful biopharmaceutical companies focusing on discovery and development of TLR7 agonists for treating liver diseases, Gilead has the most advanced TLR7 agonist pipeline with leading compounds such as GS-9620 which has entered into Phase II studies. Gilead compound GS-disclosed in US20100143301 as example 49, compound S-2 and compound S-3 disclosed in JP1999193282 were all chosen as the reference compounds in this application:
H
N)1\1e0 NN
II
ONN) ON\ 1 I 0 N
SNµN
. NO 0 CI
(GS-9620), *
H
N.----"No ,, j........N
S N
N, it (S-3).
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof, for use in the treatment or prophylaxis of liver cancer.
In some embodiments, compounds of present invention were tested and compared with the following reference compounds. As the most successful biopharmaceutical companies focusing on discovery and development of TLR7 agonists for treating liver diseases, Gilead has the most advanced TLR7 agonist pipeline with leading compounds such as GS-9620 which has entered into Phase II studies. Gilead compound GS-disclosed in US20100143301 as example 49, compound S-2 and compound S-3 disclosed in JP1999193282 were all chosen as the reference compounds in this application:
H
N)1\1e0 NN
II
ONN) ON\ 1 I 0 N
SNµN
. NO 0 CI
(GS-9620), *
H
N.----"No ,, j........N
S N
N, it (S-3).
- 20 -SYNTHESIS
The compounds of the present invention can be prepared by any conventional means.
Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the examples. All substituents, in particular, Rl to le are as defined above unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in organic chemistry.
Scheme 1 o s N C*
H
R.,. ...,C N N
+ _3p. ______________________________ 31 I\V H2 N.. N/0 R
VII VI
H i m v l'N - / \ )....,. EN1 N =-= j-"N
, 1 H S), I 0 ¨a ....1 0 _3,.. ,:z, .....,, N.,,,-.... N
N.------N S , 1 1\1 s -----N \R2 I 1 \ 2 1 \R 2 R1 R
R
V IV III
NH NH2 .......IR
H
N----"N
NH I Iõ....-- 0 11''S=== --N
as. 0 ..... 11'S=== ,...---- N 0..._---s N
I
¨S N \ 2 1 \ 2 R R
R
II I
A compound of formula VI is prepared by cyclization of isocyanate VII with aminomalononitrile p-toluenesulfonate. Then bicycle V is synthesized by reaction of compound of formula VI with benzoyl isothiocyanate in the presence of inorganic base, such as NaOH or KOH. Alkylation of bicycle V with alkylhalide in the presence of base, such as K2CO3, NaH or Cs2CO3, gives compound of formula IV. Compound of formula III
The compounds of the present invention can be prepared by any conventional means.
Suitable processes for synthesizing these compounds as well as their starting materials are provided in the schemes below and in the examples. All substituents, in particular, Rl to le are as defined above unless otherwise indicated. Furthermore, and unless explicitly otherwise stated, all reactions, reaction conditions, abbreviations and symbols have the meanings well known to a person of ordinary skill in organic chemistry.
Scheme 1 o s N C*
H
R.,. ...,C N N
+ _3p. ______________________________ 31 I\V H2 N.. N/0 R
VII VI
H i m v l'N - / \ )....,. EN1 N =-= j-"N
, 1 H S), I 0 ¨a ....1 0 _3,.. ,:z, .....,, N.,,,-.... N
N.------N S , 1 1\1 s -----N \R2 I 1 \ 2 1 \R 2 R1 R
R
V IV III
NH NH2 .......IR
H
N----"N
NH I Iõ....-- 0 11''S=== --N
as. 0 ..... 11'S=== ,...---- N 0..._---s N
I
¨S N \ 2 1 \ 2 R R
R
II I
A compound of formula VI is prepared by cyclization of isocyanate VII with aminomalononitrile p-toluenesulfonate. Then bicycle V is synthesized by reaction of compound of formula VI with benzoyl isothiocyanate in the presence of inorganic base, such as NaOH or KOH. Alkylation of bicycle V with alkylhalide in the presence of base, such as K2CO3, NaH or Cs2CO3, gives compound of formula IV. Compound of formula III
-21 -is prepared by oxidation of compound of formula IV with an oxidant, such as meta-chloroperoxybenzoic acid, urea-hydrogen peroxide adduct and HI04. Compound of formula II is obtained by imination of compound of formula III with imination reagent, such as sodium azide in acid, said acid is, for example, Eaton's reagent or PPA. Compound of formula I is obtained by reaction of compound of formula II with carbamoyl chloride in the presence of a mixed base such as pyridine and triethylamine, pyridine and DIPEA, DMAP and triethylamine, or DMAP and DIPEA.
Scheme 2 CI
,R2 ci o CI 0 I-12 N II+ N NH2 II, N r\j0- I
X" ). S ,R2 N I\I
Ri R x XI
CI
,PMB
NH2 H NI' H
NJN I
INXI\II
S NX N\ 2 S N N\ R 2 R
R iv VIII
/ R IVa H H
NjXN NJN
IIIFI) I
oS)N N Ozzs NX N
I \ 2 I R1 \R2 Ri R
III II
Compound of formula II can also be prepared as Scheme 2.
A compound of formula X is prepared by reaction of compound of formula XI with R2NH2. Reduction of compound X with reducing reagent, such as Zinc or Iron powder in AcOH, gives the compound of formula IX. Cyclization of compound of formula IX
with cyclization reagents, such as phosgene, carbonyl diimidazole, diethyl carbonate and triphosgene, affords compound of formula VIII. A compound of formula IVa is prepared by treating the compound of formula VIII with PMBNH2. A compound of formula III is prepared by deprotection of compound of formula IVa with acid, such as CF3COOH, followed by oxidation with an oxidant, such as meta-chloroperoxybenzoic acid, urea-
Scheme 2 CI
,R2 ci o CI 0 I-12 N II+ N NH2 II, N r\j0- I
X" ). S ,R2 N I\I
Ri R x XI
CI
,PMB
NH2 H NI' H
NJN I
INXI\II
S NX N\ 2 S N N\ R 2 R
R iv VIII
/ R IVa H H
NjXN NJN
IIIFI) I
oS)N N Ozzs NX N
I \ 2 I R1 \R2 Ri R
III II
Compound of formula II can also be prepared as Scheme 2.
A compound of formula X is prepared by reaction of compound of formula XI with R2NH2. Reduction of compound X with reducing reagent, such as Zinc or Iron powder in AcOH, gives the compound of formula IX. Cyclization of compound of formula IX
with cyclization reagents, such as phosgene, carbonyl diimidazole, diethyl carbonate and triphosgene, affords compound of formula VIII. A compound of formula IVa is prepared by treating the compound of formula VIII with PMBNH2. A compound of formula III is prepared by deprotection of compound of formula IVa with acid, such as CF3COOH, followed by oxidation with an oxidant, such as meta-chloroperoxybenzoic acid, urea-
- 22 -hydrogen peroxide adduct and HI04. Compound of formula II is obtained by the imination of compound of formula III with imination reagent, such as sodium azide in acid, said acid is for example Eaton's reagent or PPA.
Also described is a process for the preparation of a compound of formula (I) comprising the reaction of:
the reaction of a compound of formula (II), H
N------N> 0 N
NH
----S
11 \R2 R (II), with carbamoyl chloride in the presence of a mixed base;
wherein Rl and R2 are defined above.
In above step, the mixed base can be, for example, pyridine and triethylamine, pyridine and DIPEA, DMAP and triethylamine, or DMAP and DIPEA.
A compound of formula (I) when manufactured according to the above process, for use in the treatment or prophylaxis of liver cancer is also an object of the invention.
PHARMACEUTICAL COMPOSITIONS AND ADMINISTRATION
Another embodiment provides pharmaceutical compositions or medicaments, for use in the treatment or prophylaxis of liver cancer containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) are formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of
Also described is a process for the preparation of a compound of formula (I) comprising the reaction of:
the reaction of a compound of formula (II), H
N------N> 0 N
NH
----S
11 \R2 R (II), with carbamoyl chloride in the presence of a mixed base;
wherein Rl and R2 are defined above.
In above step, the mixed base can be, for example, pyridine and triethylamine, pyridine and DIPEA, DMAP and triethylamine, or DMAP and DIPEA.
A compound of formula (I) when manufactured according to the above process, for use in the treatment or prophylaxis of liver cancer is also an object of the invention.
PHARMACEUTICAL COMPOSITIONS AND ADMINISTRATION
Another embodiment provides pharmaceutical compositions or medicaments, for use in the treatment or prophylaxis of liver cancer containing the compounds of the invention and a therapeutically inert carrier, diluent or excipient, as well as methods of using the compounds of the invention to prepare such compositions and medicaments. In one example, compounds of formula (I) may be formulated by mixing at ambient temperature at the appropriate pH, and at the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a galenical administration form. The pH of the formulation depends mainly on the particular use and the concentration of compound, but preferably ranges anywhere from about 3 to about 8. In one example, a compound of formula (I) are formulated in an acetate buffer, at pH 5. In another embodiment, the compounds of
- 23 -formula (I) are sterile. The compound may be stored, for example, as a solid or amorphous composition, as a lyophilized formulation or as an aqueous solution.
Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The "effective amount" of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to activate TLR7 receptor and lead to produce INF-ct and other cytokines, which can be used, but not limited, for the treatment or prevention of hepatitis B and/or C viral infected patients.
In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 50 mg/kg, alternatively about 0.1 to 30 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 20 to about 1000 mg of the compound of the invention.
The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration.
Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH
modifiers, sweeteners, bulking agents, and further active agents.
A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel's
Compositions are formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners. The "effective amount" of the compound to be administered will be governed by such considerations, and is the minimum amount necessary to activate TLR7 receptor and lead to produce INF-ct and other cytokines, which can be used, but not limited, for the treatment or prevention of hepatitis B and/or C viral infected patients.
In one example, the pharmaceutically effective amount of the compound of the invention administered parenterally per dose will be in the range of about 0.1 to 50 mg/kg, alternatively about 0.1 to 30 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another embodiment, oral unit dosage forms, such as tablets and capsules, preferably contain from about 20 to about 1000 mg of the compound of the invention.
The compounds of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration.
Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.
The compounds of the present invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH
modifiers, sweeteners, bulking agents, and further active agents.
A typical formulation is prepared by mixing a compound of the present invention and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Howard C., et al., Ansel's
- 24 -Pharmaceutical Dosage Forms and Drug Delivery Systems. Philadelphia:
Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
An example of a suitable oral dosage form is a tablet containing about 20 to mg of the compound of the invention compounded with about 30 to 90 mg anhydrous lactose, about 5 to 40 mg sodium croscarmellose, about 5 to 30 mg polyvinylpyrrolidone (PVP) K30, and about 1 to 10 mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 20 to 1000 mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.
An embodiment, therefore, includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof.
In a further embodiment includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof, together with a pharmaceutically acceptable carrier or excipient.
Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof for use in the treatment of hepatitis B virus infection.
Lippincott, Williams & Wilkins, 2004; Gennaro, Alfonso R., et al. Remington: The Science and Practice of Pharmacy. Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Raymond C. Handbook of Pharmaceutical Excipients. Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to provide an elegant presentation of the drug (i.e., a compound of the present invention or pharmaceutical composition thereof) or aid in the manufacturing of the pharmaceutical product (i.e., medicament).
An example of a suitable oral dosage form is a tablet containing about 20 to mg of the compound of the invention compounded with about 30 to 90 mg anhydrous lactose, about 5 to 40 mg sodium croscarmellose, about 5 to 30 mg polyvinylpyrrolidone (PVP) K30, and about 1 to 10 mg magnesium stearate. The powdered ingredients are first mixed together and then mixed with a solution of the PVP. The resulting composition can be dried, granulated, mixed with the magnesium stearate and compressed to tablet form using conventional equipment. An example of an aerosol formulation can be prepared by dissolving the compound, for example 20 to 1000 mg, of the invention in a suitable buffer solution, e.g. a phosphate buffer, adding a tonicifier, e.g. a salt such sodium chloride, if desired. The solution may be filtered, e.g., using a 0.2 micron filter, to remove impurities and contaminants.
An embodiment, therefore, includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof.
In a further embodiment includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof, together with a pharmaceutically acceptable carrier or excipient.
Another embodiment includes a pharmaceutical composition comprising a compound of formula (I) or pharmaceutically acceptable salts or enantiomers or diastereomers thereof for use in the treatment of hepatitis B virus infection.
- 25 -INDICATIONS AND METHODS OF TREATMENT
The present invention provides methods for treating or preventing liver cancer in a patient in need thereof. In some embodiments, the liver cancer is hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer. In some embodiments, the liver cancer is a refractory cancer.
The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth/proliferation.
Examples of liver cancer include, but are not limited to, hepatocellular carcinoma, hepatoma, hepatoblastoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, sarcoma, lymphoma and hepatic angiosarcoma. In various embodiments, the liver cancer (e.g., HCC) can be intermediate, advanced, or terminal stage. The liver cancer (e.g., HCC) can be metastatic or non-metastatic. The liver cancer (e.g., HCC) can be resectable or unresectable. The liver cancer (e.g., HCC) can comprise a single tumor, multiple tumors, or a poorly defined tumor with an infiltrative growth pattern (into portal veins or hepatic veins). The liver cancer (e.g., HCC) can comprise a fibrolamellar, pseudoglandular (adenoid), pleomorphic (giant cell), or clear cell pattern. The liver cancer (e.g., HCC) can comprise a well differentiated form, and tumor cells resemble hepatocytes, form trabeculae, cords, and nests, and/or contain bile pigment in cytoplasm. The liver cancer (e.g., HCC) can comprise a poorly differentiated form, and malignant epithelial cells are discohesive, pleomorphic, anaplastic, and/or giant. In some embodiments, the liver cancer (e.g., HCC) is associated with hepatits B, hepatitis C, cirhhosis, or type 2 diabetes. The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders that are associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.
In one embodiment of the present invention the compounds (and pharmaceutical compositions and medicaments thereof) described herein are used in the prophylaxis/prevention of liver cancer in patients which have a high risk of developing liver cancer.
In one preferred embodiment of the invention the compounds described herein are especially useful as prodrugs which are converted into the active drug predominantly in the liver. One embodiment of the invention embodiment are the prodrug compounds described
The present invention provides methods for treating or preventing liver cancer in a patient in need thereof. In some embodiments, the liver cancer is hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer. In some embodiments, the liver cancer is a refractory cancer.
The terms "cancer" and "cancerous" refer to or describe the physiological condition in mammals that is typically characterized by unregulated cell growth/proliferation.
Examples of liver cancer include, but are not limited to, hepatocellular carcinoma, hepatoma, hepatoblastoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, sarcoma, lymphoma and hepatic angiosarcoma. In various embodiments, the liver cancer (e.g., HCC) can be intermediate, advanced, or terminal stage. The liver cancer (e.g., HCC) can be metastatic or non-metastatic. The liver cancer (e.g., HCC) can be resectable or unresectable. The liver cancer (e.g., HCC) can comprise a single tumor, multiple tumors, or a poorly defined tumor with an infiltrative growth pattern (into portal veins or hepatic veins). The liver cancer (e.g., HCC) can comprise a fibrolamellar, pseudoglandular (adenoid), pleomorphic (giant cell), or clear cell pattern. The liver cancer (e.g., HCC) can comprise a well differentiated form, and tumor cells resemble hepatocytes, form trabeculae, cords, and nests, and/or contain bile pigment in cytoplasm. The liver cancer (e.g., HCC) can comprise a poorly differentiated form, and malignant epithelial cells are discohesive, pleomorphic, anaplastic, and/or giant. In some embodiments, the liver cancer (e.g., HCC) is associated with hepatits B, hepatitis C, cirhhosis, or type 2 diabetes. The terms "cell proliferative disorder" and "proliferative disorder" refer to disorders that are associated with some degree of abnormal cell proliferation. In one embodiment, the cell proliferative disorder is cancer.
In one embodiment of the present invention the compounds (and pharmaceutical compositions and medicaments thereof) described herein are used in the prophylaxis/prevention of liver cancer in patients which have a high risk of developing liver cancer.
In one preferred embodiment of the invention the compounds described herein are especially useful as prodrugs which are converted into the active drug predominantly in the liver. One embodiment of the invention embodiment are the prodrug compounds described
- 26 -herein for use in the treatment of liver cancer wherein the compounds are prodrugs of the formula (I), N H 2 .....¨R3 N===="" N
H N >0 II,00..--, .0õ...---,N
0=S N
1 1 \R2 R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, C1-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, C1_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
H N >0 II,00..--, .0õ...---,N
0=S N
1 1 \R2 R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, C1-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, C1_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
-27 -6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, and wherein the prodrug compounds of formula I are converted in the human liver into the active drug of the formula II
H
N> NH 0 0.j IN/..-------N
------S
11 \R2 R (II), wherein Rl and R2 are defined above.
Exemplary conversion ratios using human liver microsomes are shown in Example 50.
Also example 61 demonstrates the liver as the primary site of conversion of the prodrug into its active form.
One preferred embodiment of the invention are the (prodrug) compounds described herein wherein the compounds are susceptible for the conversion into its active form by the liver enzymes CYP2C9 and CYP2C19. One preferred embodiment of the invention are the (prodrug) compounds described herein wherein the compounds show a conversion rate into the active compound of >10 nmol/min/mg protein in human hepatocytes and of <2 nmol/min/mg protein in human enterocytes (as measured in an appropriate assay using human hepatocytes and human enterocytes.
COMBINATION TREATMENT
One aspect of the invention is the combined treatment (combination treatment) of a patient suffering from liver cancer with the compound of formula I with
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, and wherein the prodrug compounds of formula I are converted in the human liver into the active drug of the formula II
H
N> NH 0 0.j IN/..-------N
------S
11 \R2 R (II), wherein Rl and R2 are defined above.
Exemplary conversion ratios using human liver microsomes are shown in Example 50.
Also example 61 demonstrates the liver as the primary site of conversion of the prodrug into its active form.
One preferred embodiment of the invention are the (prodrug) compounds described herein wherein the compounds are susceptible for the conversion into its active form by the liver enzymes CYP2C9 and CYP2C19. One preferred embodiment of the invention are the (prodrug) compounds described herein wherein the compounds show a conversion rate into the active compound of >10 nmol/min/mg protein in human hepatocytes and of <2 nmol/min/mg protein in human enterocytes (as measured in an appropriate assay using human hepatocytes and human enterocytes.
COMBINATION TREATMENT
One aspect of the invention is the combined treatment (combination treatment) of a patient suffering from liver cancer with the compound of formula I with
- 28 -Surprisingly, we found that that a combination therapy of the compounds of formula I and an anti-PD-Ll/PD1 axis treatment is highly effective for liver tumors Therefore one aspect of the invention is a compound of the formula (I) (or a medicament or a pharmaceutical composition comprising such compound), N H 2 ....¨R3 N--'N> 0 11.õ....... ------....N
0=S N
11 \ 2 R
R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6a1ky1, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzyl-2-(propylsulfonimidoy1)-7-(pyrrolidine- 1 -carb onyl)purin-8-one ;
6-amino-9-benzyl-7-(piperidine- 1 -c arb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(3 ,3 -dimethylpyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbonyl)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
0=S N
11 \ 2 R
R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6a1ky1, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzyl-2-(propylsulfonimidoy1)-7-(pyrrolidine- 1 -carb onyl)purin-8-one ;
6-amino-9-benzyl-7-(piperidine- 1 -c arb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(3 ,3 -dimethylpyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbonyl)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
- 29 -6-amino-9-benzyl-7-(2-oxa-6-azaspiro [3.3 ] heptane- 6-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(3 ,3 -difluoropyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(3 -fluoro-3 -methyl-pyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, for use in a) the treatment of liver cancer in combination with an antagonistic PD1 or antagonistic PD-Li antibodyõ
or b) the treatment of a patient suffering from liver cancer in combination with an antagonistic PD1 or antagonistic PD-Li antibody.
One embodiment of the invention is a compound of the formula (I) (or a medicament or a pharmaceutical composition comprising such compound), N H2 ....---R
N----N> 0 0=SN---------N
11 y R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6allcoxyCi_6alkyl;
R5 is (Ci_6alkyl)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci_ 6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_
6-amino-9-benzyl-7-(3 ,3 -difluoropyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(3 -fluoro-3 -methyl-pyrrolidine- 1 -carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, for use in a) the treatment of liver cancer in combination with an antagonistic PD1 or antagonistic PD-Li antibodyõ
or b) the treatment of a patient suffering from liver cancer in combination with an antagonistic PD1 or antagonistic PD-Li antibody.
One embodiment of the invention is a compound of the formula (I) (or a medicament or a pharmaceutical composition comprising such compound), N H2 ....---R
N----N> 0 0=SN---------N
11 y R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6allcoxyCi_6alkyl;
R5 is (Ci_6alkyl)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci_ 6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)C1_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_
- 30 -6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyCi_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro [3.3] heptane-6-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3-fluoro-3-methyl-pyrro lidine-l-c arb ony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, for use in the treatment or or prophylaxis of liver cancer wherein an antagonistic PD1 or antagonistic PD-Li antibody is co-administered (wherein the treatment is in combination with an antagonistic PD1 or antagonistic PD-Li antibody).
One embodiment of the invention is the use of a compound of the formula (I),
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro [3.3] heptane-6-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3-fluoro-3-methyl-pyrro lidine-l-c arb ony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded, for use in the treatment or or prophylaxis of liver cancer wherein an antagonistic PD1 or antagonistic PD-Li antibody is co-administered (wherein the treatment is in combination with an antagonistic PD1 or antagonistic PD-Li antibody).
One embodiment of the invention is the use of a compound of the formula (I),
-31 -N H 2 .....-R3 N----"N
H N
" 'N
0=S N
1 1 \ 2 R
R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, C1-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, C1_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, C1_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(2-oxa-6-azaspiro [3.3] heptane-6-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
H N
" 'N
0=S N
1 1 \ 2 R
R (I), wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, C1-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, C1_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, C1_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzyl-7-(2-oxa-6-azaspiro [3.3] heptane-6-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
32 and their enantiomers or diastereomers are excluded, for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein an antagonistic PD1 or antagonistic PD-Li antibody is co-administered (wherein the treatment is in combination with an antagonistic PD1 or antagonistic PD-Li antibody).
In another embodiment of present invention, the particular compounds of formula (I) which are used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody are selected from the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-pip eridyl)piperidine-l-c arb onyl] -2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-l-c arb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
In another embodiment of present invention, the particular compounds of formula (I) which are used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody are selected from the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-pip eridyl)piperidine-l-c arb onyl] -2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-l-c arb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
- 33 -6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 34 -Methyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl] -N-methyl-carbamate;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine- 1 -carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine- 1-carbonyl)purin-8-one;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine- 1 -carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine- 1-carbonyl)purin-8-one;
- 35 -6-Amino-2-[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
- 36 -6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
In another embodiment of present invention, the particular compounds of formula (I) which are used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody are selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
In another embodiment of present invention, the particular compounds of formula (I) which are used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody are selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
- 37 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
In another embodiment of present invention, the particular compound of formula (I) which is used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody is: 6-Amino-9-[(4-chlorophenyOmethy1]-N-ethy1-2[S(5)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide In one embodiment the co-administration (or combination therapy or treatment in combination with or combination treatment) of the compound of formula I and the antagonistic PD1 or antagonistic PD-Li antibody is simultaneously. In one embodiment the co-administration (or combination therapy or treatment in combination with or combination treatment) of the compound of formula I and the antagonistic PD1 or antagonistic PD-Li antibody is sequentially.
The terms "administered in combination with" or "co-administration", "co-administering", "combination therapy", "treatment in combination with "or "combination treatment" refer to the administration of the Compound of formula I as described herein, and the antagonistic PD1 or PD-Li antibody, as described herein e.g. as separate formulations/applications (or as one single formulation/application). The co-administration can be simultaneous or sequential in either order, wherein there is a time period while both (or all) active agents simultaneously exert their biological activities. The co-administration is either simultaneously or sequentially (e.g. intravenous (i.v.) through a continuous infusion. In one embodiment the co-administration is simultaneously. In one embodiment the co-administration is sequentially. The co-administration is either simultaneously or sequentially (e.g. intravenous (i.v.) through a continuous infusion.
It is self-evident that the antibodies are administered to the patient in a "therapeutically effective amount" (or simply "effective amount") which is the amount of the respective compound or combination that will elicit the biological or medical response of a tissue,
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
In another embodiment of present invention, the particular compound of formula (I) which is used in the combination therapy with the antagonistic PD1 or antagonistic PD-Li antibody is: 6-Amino-9-[(4-chlorophenyOmethy1]-N-ethy1-2[S(5)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide In one embodiment the co-administration (or combination therapy or treatment in combination with or combination treatment) of the compound of formula I and the antagonistic PD1 or antagonistic PD-Li antibody is simultaneously. In one embodiment the co-administration (or combination therapy or treatment in combination with or combination treatment) of the compound of formula I and the antagonistic PD1 or antagonistic PD-Li antibody is sequentially.
The terms "administered in combination with" or "co-administration", "co-administering", "combination therapy", "treatment in combination with "or "combination treatment" refer to the administration of the Compound of formula I as described herein, and the antagonistic PD1 or PD-Li antibody, as described herein e.g. as separate formulations/applications (or as one single formulation/application). The co-administration can be simultaneous or sequential in either order, wherein there is a time period while both (or all) active agents simultaneously exert their biological activities. The co-administration is either simultaneously or sequentially (e.g. intravenous (i.v.) through a continuous infusion. In one embodiment the co-administration is simultaneously. In one embodiment the co-administration is sequentially. The co-administration is either simultaneously or sequentially (e.g. intravenous (i.v.) through a continuous infusion.
It is self-evident that the antibodies are administered to the patient in a "therapeutically effective amount" (or simply "effective amount") which is the amount of the respective compound or combination that will elicit the biological or medical response of a tissue,
- 38 -system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician.
The amount of co-administration and the timing of co-administration will depend on the type (species, gender, age, weight, etc.) and condition of the patient being treated and the severity of the disease or condition being treated. Said compounds of formula I and said antibodies are suitably co-administered to the patient at one time or over a series of treatments e.g. on the same day or on the day after.
PD-1/PD-Li/PD-L2 pathway:
An important negative co-stimulatory signal regulating T cell activation is provided by programmed death ¨ 1 receptor (PD-1)(CD279), and its ligand binding partners PD-Li (B7-H1, CD274; SEQ ID NO: 13) and PD-L2 (B7-DC, CD273). The negative regulatory role of PD-1 was revealed by PD-1 knock outs (Pdcd1-/-), which are prone to autoimmunity. Nishimura et al., Immunity 11: 141-51 (1999); Nishimura et al., Science 291: 319-22 (2001). PD-1 is related to CD28 and CTLA-4, but lacks the membrane proximal cysteine that allows homodimerization. The cytoplasmic domain of PD-1 contains an immunoreceptor tyrosine-based inhibition motif (ITIM, V/IxYxxL/V).
only binds to PD-Li and PD-L2. Freeman et al., J. Exp. Med. 192: 1-9 (2000);
Dong et al., Nature Med. 5: 1365-1369 (1999); Latchman et al., Nature Immunol. 2: 261-268 (2001);
Tseng et al., J. Exp. Med. 193: 839-846 (2001).
PD-1 can be expressed on T cells, B cells, natural killer T cells, activated monocytes and dendritic cells (DCs). PD-1 is expressed by activated, but not by unstimulated human CD4+ and CD8+ T cells, B cells and myeloid cells. This stands in contrast to the more restricted expression of CD28 and CTLA-4. Nishimura et al., Int. Immunol. 8:
(1996); Boettler et al., J. Virol. 80: 3532-40 (2006). There are at least 4 variants of PD-1 that have been cloned from activated human T cells, including transcripts lacking (i) exon 2, (ii) exon 3, (iii) exons 2 and 3 or (iv) exons 2 through 4. Nielsen et al., Cell. Immunol.
235: 109-16 (2005). With the exception of PD-ldeltaex3, all variants are expressed at similar levels as full length PD-1 in resting peripheral blood mononuclear cells (PBMCs).
Expression of all variants is significantly induced upon activation of human T
cells with anti-CD3 and anti-CD28. The PD-1 de1taex3 variants lacks a transmembrane domain, and resembles soluble CTLA-4, which plays an important role in autoimmunity. Ueda et al.,
The amount of co-administration and the timing of co-administration will depend on the type (species, gender, age, weight, etc.) and condition of the patient being treated and the severity of the disease or condition being treated. Said compounds of formula I and said antibodies are suitably co-administered to the patient at one time or over a series of treatments e.g. on the same day or on the day after.
PD-1/PD-Li/PD-L2 pathway:
An important negative co-stimulatory signal regulating T cell activation is provided by programmed death ¨ 1 receptor (PD-1)(CD279), and its ligand binding partners PD-Li (B7-H1, CD274; SEQ ID NO: 13) and PD-L2 (B7-DC, CD273). The negative regulatory role of PD-1 was revealed by PD-1 knock outs (Pdcd1-/-), which are prone to autoimmunity. Nishimura et al., Immunity 11: 141-51 (1999); Nishimura et al., Science 291: 319-22 (2001). PD-1 is related to CD28 and CTLA-4, but lacks the membrane proximal cysteine that allows homodimerization. The cytoplasmic domain of PD-1 contains an immunoreceptor tyrosine-based inhibition motif (ITIM, V/IxYxxL/V).
only binds to PD-Li and PD-L2. Freeman et al., J. Exp. Med. 192: 1-9 (2000);
Dong et al., Nature Med. 5: 1365-1369 (1999); Latchman et al., Nature Immunol. 2: 261-268 (2001);
Tseng et al., J. Exp. Med. 193: 839-846 (2001).
PD-1 can be expressed on T cells, B cells, natural killer T cells, activated monocytes and dendritic cells (DCs). PD-1 is expressed by activated, but not by unstimulated human CD4+ and CD8+ T cells, B cells and myeloid cells. This stands in contrast to the more restricted expression of CD28 and CTLA-4. Nishimura et al., Int. Immunol. 8:
(1996); Boettler et al., J. Virol. 80: 3532-40 (2006). There are at least 4 variants of PD-1 that have been cloned from activated human T cells, including transcripts lacking (i) exon 2, (ii) exon 3, (iii) exons 2 and 3 or (iv) exons 2 through 4. Nielsen et al., Cell. Immunol.
235: 109-16 (2005). With the exception of PD-ldeltaex3, all variants are expressed at similar levels as full length PD-1 in resting peripheral blood mononuclear cells (PBMCs).
Expression of all variants is significantly induced upon activation of human T
cells with anti-CD3 and anti-CD28. The PD-1 de1taex3 variants lacks a transmembrane domain, and resembles soluble CTLA-4, which plays an important role in autoimmunity. Ueda et al.,
- 39 -Nature 423: 506-11 (2003). This variant is enriched in the synovial fluid and sera of patients with rheumatoid arthritis. Wan et al., J. Immunol. 177: 8844-50 (2006).
The two PD-1 ligands differ in their expression patterns. PD-Li is constitutively expressed on mouse T and B cells, CDs, macrophages, mesenchymal stem cells and bone marrow-derived mast cells. Yamazaki et al., J. Immunol. 169: 5538-45 (2002). PD-Li is expressed on a wide range of nonhematopoietic cells (e.g., cornea, lung, vascular epithelium, liver nonparenchymal cells, mesenchymal stem cells, pancreatic islets, placental synctiotrophoblasts, keratinocytes, etc.) [Keir et al., Annu. Rev. Immunol.
26: 677-704 (2008)], and is upregulated on a number of cell types after activation. Both type I and type II interferons IFN's) upregulate PD-Li. Eppihimer et al., Microcirculation 9:
(2002); Schreiner et al., J. Neuroimmunol. 155: 172-82 (2004). PD-Li expression in cell lines is decreased when MyD88, TRAF6 and MEK are inhibited. Liu et al., Blood 110:
296-304 (2007). JAK2 has also been implicated in PD-Li induction. Lee et al., FEBS Lett.
580: 755-62 (2006); Liu et al., Blood 110: 296-304 (2007). Loss or inhibition of phosphatase and tensin homolog (PTEN), a cellular phosphatase that modified phosphatidylinositol 3-kinase (PI3K) and Akt signaling, increased post-transcriptional PD-Li expression in cancers. Parsa et al., Nat. Med. 13: 84-88 (2007).
PD-L2 expression is more restricted than PD-Li. PD-L2 is inducibly expressed on DCs, macrophages, and bone marrow-derived mast cells. PD-L2 is also expressed on about half to two-thirds of resting peritoneal B1 cells, but not on conventional B2 B
cells. Zhong et al., Eur. J. Immunol. 37: 2405-10 (2007). PD-L2+ B1 cells bind phosphatidylcholine and may be important for innate immune responses against bacterial antigens.
Induction of PD-L2 by IFN-gamma is partially dependent upon NF-kappaB. Liang et al., Eur. J.
Immunol.
33: 2706-16 (2003). PD-L2 can also be induced on monocytes and macrophages by GM-CF, IL-4 and IFN-gamma. Yamazaki et al., J. Immunol. 169: 5538-45 (2002); Loke et al., PNAS 100:5336-41 (2003).
PD-1 signaling typically has a greater effect on cytokine production than on cellular proliferation, with significant effects on IFN-gamma, TNF-alpha and IL-2 production. PD-1 mediated inhibitory signaling also depends on the strength of the TCR
signaling, with greater inhibition delivered at low levels of TCR stimulation. This reduction can be overcome by costimulation through CD28 [Freeman et al., J. Exp. Med. 192: 1027-(2000)] or the presence of IL-2 [Carter et al., Eur. J. Immunol. 32: 634-43 (2002)].
The two PD-1 ligands differ in their expression patterns. PD-Li is constitutively expressed on mouse T and B cells, CDs, macrophages, mesenchymal stem cells and bone marrow-derived mast cells. Yamazaki et al., J. Immunol. 169: 5538-45 (2002). PD-Li is expressed on a wide range of nonhematopoietic cells (e.g., cornea, lung, vascular epithelium, liver nonparenchymal cells, mesenchymal stem cells, pancreatic islets, placental synctiotrophoblasts, keratinocytes, etc.) [Keir et al., Annu. Rev. Immunol.
26: 677-704 (2008)], and is upregulated on a number of cell types after activation. Both type I and type II interferons IFN's) upregulate PD-Li. Eppihimer et al., Microcirculation 9:
(2002); Schreiner et al., J. Neuroimmunol. 155: 172-82 (2004). PD-Li expression in cell lines is decreased when MyD88, TRAF6 and MEK are inhibited. Liu et al., Blood 110:
296-304 (2007). JAK2 has also been implicated in PD-Li induction. Lee et al., FEBS Lett.
580: 755-62 (2006); Liu et al., Blood 110: 296-304 (2007). Loss or inhibition of phosphatase and tensin homolog (PTEN), a cellular phosphatase that modified phosphatidylinositol 3-kinase (PI3K) and Akt signaling, increased post-transcriptional PD-Li expression in cancers. Parsa et al., Nat. Med. 13: 84-88 (2007).
PD-L2 expression is more restricted than PD-Li. PD-L2 is inducibly expressed on DCs, macrophages, and bone marrow-derived mast cells. PD-L2 is also expressed on about half to two-thirds of resting peritoneal B1 cells, but not on conventional B2 B
cells. Zhong et al., Eur. J. Immunol. 37: 2405-10 (2007). PD-L2+ B1 cells bind phosphatidylcholine and may be important for innate immune responses against bacterial antigens.
Induction of PD-L2 by IFN-gamma is partially dependent upon NF-kappaB. Liang et al., Eur. J.
Immunol.
33: 2706-16 (2003). PD-L2 can also be induced on monocytes and macrophages by GM-CF, IL-4 and IFN-gamma. Yamazaki et al., J. Immunol. 169: 5538-45 (2002); Loke et al., PNAS 100:5336-41 (2003).
PD-1 signaling typically has a greater effect on cytokine production than on cellular proliferation, with significant effects on IFN-gamma, TNF-alpha and IL-2 production. PD-1 mediated inhibitory signaling also depends on the strength of the TCR
signaling, with greater inhibition delivered at low levels of TCR stimulation. This reduction can be overcome by costimulation through CD28 [Freeman et al., J. Exp. Med. 192: 1027-(2000)] or the presence of IL-2 [Carter et al., Eur. J. Immunol. 32: 634-43 (2002)].
- 40 -Evidence is mounting that signaling through PD-Li and PD-L2 may be bidirectional. That is, in addition to modifying TCR or BCR signaling, signaling may also be delivered back to the cells expressing PD-Li and PD-L2. While treatment of dendritic cells with a naturally human anti-PD-L2 antibody isolated from a patient with Waldenstrom's macroglobulinemia was not found to upregulate MHC II or B7 costimulatory molecules, such cells did produce greater amount of proinflammatory cytokines, particularly TNF-alpha and IL-6, and stimulated T cell proliferation. Nguyen et al., J. Exp.
Med. 196: 1393-98 (2002). Treatment of mice with this antibody also (1) enhanced resistance to transplanted b16 melanoma and rapidly induced tumor-specific CTL.
Radhakrishnan et al., J. Immunol. 170: 1830-38 (2003); Radhakrishnan et al., Cancer Res. 64: 4965-72 (2004);
Heckman et al., Eur. J. Immunol. 37: 1827-35 (2007); (2) blocked development of airway inflammatory disease in a mouse model of allergic asthma. Radhakrishnan et al., J.
Immunol. 173: 1360-65 (2004); Radhakrishnan et al., J. Allergy Clin. Immunol.
116: 668-74 (2005).
Further evidence of reverse signaling into dendritic cells ("DC's") results from studies of bone marrow derived DC's cultured with soluble PD-1 (PD-1 EC domain fused to Ig constant region ¨ "s-PD-1"). Kuipers et al., Eur. J. Immunol. 36: 2472-82 (2006). This sPD-1 inhibited DC activation and increased IL-10 production, in a manner reversible through administration of anti-PD-1.
Additionally, several studies show a receptor for PD-Li or PD-L2 that is independent of PD-1. B7.1 has already been identified as a binding partner for PD-Li. Butte et al., Immunity 27: 111-22 (2007). Chemical crosslinking studies suggest that PD-Li and B7.1 can interact through their IgV-like domains. B7.1:PD-L1 interactions can induce an inhibitory signal into T cells. Ligation of PD-Li on CD4+ T cells by B7.1 or ligation of B7.1 on CD4+ T cells by PD-Li delivers an inhibitory signal. T cells lacking CD28 and CTLA-4 show decreased proliferation and cytokine production when stimulated by anti-CD3 plus B7.1 coated beads. In T cells lacking all the receptors for B7.1 (i.e., CD28, CTLA-4 and PD-L1), T cell proliferation and cytokine production were no longer inhibited by anti-CD3 plus B7.1 coated beads. This indicates that B7.1 acts specifically through PD-Li on the T-cell in the absence of CD28 and CTLA-4. Similarly, T cells lacking showed decreased proliferation and cytokine production when stimulated in the presence of anti-CD3 plus PD-Li coated beads, demonstrating the inhibitory effect of PD-Li ligation on B7.1 on T cells. When T cells lacking all known receptors for PD-Li (i.e., no
Med. 196: 1393-98 (2002). Treatment of mice with this antibody also (1) enhanced resistance to transplanted b16 melanoma and rapidly induced tumor-specific CTL.
Radhakrishnan et al., J. Immunol. 170: 1830-38 (2003); Radhakrishnan et al., Cancer Res. 64: 4965-72 (2004);
Heckman et al., Eur. J. Immunol. 37: 1827-35 (2007); (2) blocked development of airway inflammatory disease in a mouse model of allergic asthma. Radhakrishnan et al., J.
Immunol. 173: 1360-65 (2004); Radhakrishnan et al., J. Allergy Clin. Immunol.
116: 668-74 (2005).
Further evidence of reverse signaling into dendritic cells ("DC's") results from studies of bone marrow derived DC's cultured with soluble PD-1 (PD-1 EC domain fused to Ig constant region ¨ "s-PD-1"). Kuipers et al., Eur. J. Immunol. 36: 2472-82 (2006). This sPD-1 inhibited DC activation and increased IL-10 production, in a manner reversible through administration of anti-PD-1.
Additionally, several studies show a receptor for PD-Li or PD-L2 that is independent of PD-1. B7.1 has already been identified as a binding partner for PD-Li. Butte et al., Immunity 27: 111-22 (2007). Chemical crosslinking studies suggest that PD-Li and B7.1 can interact through their IgV-like domains. B7.1:PD-L1 interactions can induce an inhibitory signal into T cells. Ligation of PD-Li on CD4+ T cells by B7.1 or ligation of B7.1 on CD4+ T cells by PD-Li delivers an inhibitory signal. T cells lacking CD28 and CTLA-4 show decreased proliferation and cytokine production when stimulated by anti-CD3 plus B7.1 coated beads. In T cells lacking all the receptors for B7.1 (i.e., CD28, CTLA-4 and PD-L1), T cell proliferation and cytokine production were no longer inhibited by anti-CD3 plus B7.1 coated beads. This indicates that B7.1 acts specifically through PD-Li on the T-cell in the absence of CD28 and CTLA-4. Similarly, T cells lacking showed decreased proliferation and cytokine production when stimulated in the presence of anti-CD3 plus PD-Li coated beads, demonstrating the inhibitory effect of PD-Li ligation on B7.1 on T cells. When T cells lacking all known receptors for PD-Li (i.e., no
-41 -PD-1 and B7.1), T cell proliferation was no longer impaired by anti-CD3 plus PD-Li coated beads. Thus, PD-Li can exert an inhibitory effect on T cells either through B7.1 or PD-1.
The direct interaction between B7.1 and PD-Li suggests that the current understanding of costimulation is incomplete, and underscores the significance to the expression of these molecules on T cells. Studies of PD-L1-/- T cells indicate that PD-Li on T
cells can downregulate T cell cytokine production. Latchman et al., Proc. Natl. Acad.
Sci. USA 101:
10691-96 (2004). Because both PD-Li and B7.1 are expressed on T cells, B
cells, DCs and macrophages, there is the potential for directional interactions between B7.1 and PD-Li on these cells types. Additionally, PD-Li on non-hematopoietic cells may interact with B7.1 as well as PD-1 on T cells, raising the question of whether PD-Li is involved in their regulation. One possible explanation for the inhibitory effect of B7.1:PD-L1 interaction is that T cell PD-Li may trap or segregate away APC B7.1 from interaction with CD28.
As a result, the antagonism of signaling through PD-L1, including blocking PD-Li from interacting with either PD-1, B7.1 or both, thereby preventing PD-Li from sending a negative co-stimulatory signal to T-cells and other antigen presenting cells is likely to enhance immunity in response to infection (e.g., acute and chronic) and tumor immunity.
An exemplary PD-Li antagonist is the anti-PD-Li antibody atezolizumab. Other antagonistic PD-Li antibodies are durvalumab and avelumab.
In another embodiment, the anti-PD-Ll/PD1 interaction can blocked by antagonist anti-PD-1 antibodies like the antagonistic PD1 antibodies pembrolizumab or nivolumab or an anti-PD1 antibody comprising the variable heavy chain and light chain domainss of PD1-0103-0312.
The term "human PD-Li" refers to the human protein PD-Li (SEQ ID NO: 13, PD-1 signaling typically). As used herein, "binding to human PD-Li" or "specifically binding to human PD-Li" or "which binds to human PD-Li" or "anti- PD-Li antibody" or "antagonistic PD-Li" refers to an antibody specifically binding to the human PD-Li antigen with a binding affinity of KD-value of 1.0 x 10-8 mo1/1 or lower, in one embodiment of a KD-value of 1.0 x10-9 mo1/1 or lower. The binding affinity is determined with a standard binding assay, such as surface plasmon resonance technique (BIAcore0, GE-Healthcare Uppsala, Sweden). Thus an "antibody binding to human PD-Li" as used
The direct interaction between B7.1 and PD-Li suggests that the current understanding of costimulation is incomplete, and underscores the significance to the expression of these molecules on T cells. Studies of PD-L1-/- T cells indicate that PD-Li on T
cells can downregulate T cell cytokine production. Latchman et al., Proc. Natl. Acad.
Sci. USA 101:
10691-96 (2004). Because both PD-Li and B7.1 are expressed on T cells, B
cells, DCs and macrophages, there is the potential for directional interactions between B7.1 and PD-Li on these cells types. Additionally, PD-Li on non-hematopoietic cells may interact with B7.1 as well as PD-1 on T cells, raising the question of whether PD-Li is involved in their regulation. One possible explanation for the inhibitory effect of B7.1:PD-L1 interaction is that T cell PD-Li may trap or segregate away APC B7.1 from interaction with CD28.
As a result, the antagonism of signaling through PD-L1, including blocking PD-Li from interacting with either PD-1, B7.1 or both, thereby preventing PD-Li from sending a negative co-stimulatory signal to T-cells and other antigen presenting cells is likely to enhance immunity in response to infection (e.g., acute and chronic) and tumor immunity.
An exemplary PD-Li antagonist is the anti-PD-Li antibody atezolizumab. Other antagonistic PD-Li antibodies are durvalumab and avelumab.
In another embodiment, the anti-PD-Ll/PD1 interaction can blocked by antagonist anti-PD-1 antibodies like the antagonistic PD1 antibodies pembrolizumab or nivolumab or an anti-PD1 antibody comprising the variable heavy chain and light chain domainss of PD1-0103-0312.
The term "human PD-Li" refers to the human protein PD-Li (SEQ ID NO: 13, PD-1 signaling typically). As used herein, "binding to human PD-Li" or "specifically binding to human PD-Li" or "which binds to human PD-Li" or "anti- PD-Li antibody" or "antagonistic PD-Li" refers to an antibody specifically binding to the human PD-Li antigen with a binding affinity of KD-value of 1.0 x 10-8 mo1/1 or lower, in one embodiment of a KD-value of 1.0 x10-9 mo1/1 or lower. The binding affinity is determined with a standard binding assay, such as surface plasmon resonance technique (BIAcore0, GE-Healthcare Uppsala, Sweden). Thus an "antibody binding to human PD-Li" as used
- 42 -herein refers to an antibody specifically binding to the human PD-Li antigen with a binding affinity of KD 1.0 x 10-8 mo1/1 or lower (in one embodiment 1.0 x 10-8 mo1/1- 1.0 x 10-13 mo1/1), in on embodiment of a KD 1.0 x10-9 mo1/1 or lower (in one embodiment 1.0 x 10-9 mo1/1- 1.0 x 10-13 mo1/1).
The term "human PD1" refers to the human protein PD1 (SEQ ID NO: 14, PD-1 signaling typically). As used herein, "binding to human PD 1" or "specifically binding to human PD1" or "which binds to human PD1" or "anti-PD1 antibody" or "antagonistic PD1" refers to an antibody specifically binding to the human PD1 antigen with a binding affinity of KD-value of 1.0 x 10-8 mo1/1 or lower, in one embodiment of a KD-value of 1.0 x10-9 mo1/1 or lower. The binding affinity is determined with a standard binding assay, such as surface plasmon resonance technique (BIAcore0, GE-Healthcare Uppsala, Sweden).
Thus an "antibody binding to human PD1" as used herein refers to an antibody specifically binding to the human PD1 antigen with a binding affinity of KD 1.0 x 10-8 mo1/1 or lower (in one embodiment 1.0 x 10-8 mo1/1- 1.0 x 10-13 mo1/1), in on embodiment of a KD 1.0 x10-9 mo1/1 or lower (in one embodiment 1.0 x 10-9 mo1/1- 1.0 x 10-13 mo1/1).
The "variable domain" (variable domain of a light chain (VL), variable domain of a heavy chain (VH) as used herein denotes each of the pair of light and heavy chains which is involved directly in binding the antibody to the antigen. The domains of variable human light and heavy chains have the same general structure and each domain comprises four framework (FR) regions whose sequences are widely conserved, connected by three "hypervariable regions" (or complementarity determining regions, CDRs). The framework regions adopt a I3-sheet conformation and the CDRs may form loops connecting the 13-sheet structure. The CDRs in each chain are held in their three-dimensional structure by the framework regions and form together with the CDRs from the other chain the antigen binding site. The antibody heavy and light chain CDR3 regions play a particularly important role in the binding specificity/affinity of the antibodies according to the invention and therefore provide a further object of the invention.
The term "constant region" as used within the current applications denotes the sum of the domains of an antibody other than the variable region. The constant region is not involved directly in binding of an antigen, but exhibits various effector functions.
Depending on the amino acid sequence of the constant region of their heavy chains, antibodies are divided in the classes: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses, such as IgGl, IgG2, IgG3, and IgG4, IgAl and IgA2. The heavy chain constant
The term "human PD1" refers to the human protein PD1 (SEQ ID NO: 14, PD-1 signaling typically). As used herein, "binding to human PD 1" or "specifically binding to human PD1" or "which binds to human PD1" or "anti-PD1 antibody" or "antagonistic PD1" refers to an antibody specifically binding to the human PD1 antigen with a binding affinity of KD-value of 1.0 x 10-8 mo1/1 or lower, in one embodiment of a KD-value of 1.0 x10-9 mo1/1 or lower. The binding affinity is determined with a standard binding assay, such as surface plasmon resonance technique (BIAcore0, GE-Healthcare Uppsala, Sweden).
Thus an "antibody binding to human PD1" as used herein refers to an antibody specifically binding to the human PD1 antigen with a binding affinity of KD 1.0 x 10-8 mo1/1 or lower (in one embodiment 1.0 x 10-8 mo1/1- 1.0 x 10-13 mo1/1), in on embodiment of a KD 1.0 x10-9 mo1/1 or lower (in one embodiment 1.0 x 10-9 mo1/1- 1.0 x 10-13 mo1/1).
The "variable domain" (variable domain of a light chain (VL), variable domain of a heavy chain (VH) as used herein denotes each of the pair of light and heavy chains which is involved directly in binding the antibody to the antigen. The domains of variable human light and heavy chains have the same general structure and each domain comprises four framework (FR) regions whose sequences are widely conserved, connected by three "hypervariable regions" (or complementarity determining regions, CDRs). The framework regions adopt a I3-sheet conformation and the CDRs may form loops connecting the 13-sheet structure. The CDRs in each chain are held in their three-dimensional structure by the framework regions and form together with the CDRs from the other chain the antigen binding site. The antibody heavy and light chain CDR3 regions play a particularly important role in the binding specificity/affinity of the antibodies according to the invention and therefore provide a further object of the invention.
The term "constant region" as used within the current applications denotes the sum of the domains of an antibody other than the variable region. The constant region is not involved directly in binding of an antigen, but exhibits various effector functions.
Depending on the amino acid sequence of the constant region of their heavy chains, antibodies are divided in the classes: IgA, IgD, IgE, IgG and IgM, and several of these may be further divided into subclasses, such as IgGl, IgG2, IgG3, and IgG4, IgAl and IgA2. The heavy chain constant
- 43 -regions that correspond to the different classes of antibodies are called a, 6, E, y, and , respectively. The light chain constant regions which can be found in all five antibody classes are called K (kappa) and k (lambda).
The terms "constant region derived from human origin" or "human constant region" as used in the current application denotes a constant heavy chain region of a human antibody of the subclass IgGl, IgG2, IgG3, or IgG4 and/or a constant light chain kappa or lambda region. Such constant regions are well known in the state of the art and e.g.
described by Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991) (see also e.g.
Johnson, G., and Wu, T.T., Nucleic Acids Res. 28 (2000) 214-218; Kabat, E.A., et al., Proc.
Natl. Acad. Sci.
USA 72 (1975) 2785-2788). Within the application for the numbering of positions and mutations the EU numbering system (EU Index) according to Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991) is used and referred to as "numbering according to EU Index of Kabat".
In one embodiment the antagonistic anti-PD1 antibody which binds to human PD1 used in the combination therapy described herein is nivolumab or pembrolizumab and is characterized in comprising the following VH and VL sequences as described herein:
TABLE:
anti-PD-Li antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
nivolumab 1 2 pembrolizumab 3 4 In one preferred embodiment of the invention the compound of formula I used in the combination therapy described herein is selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[5(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
The terms "constant region derived from human origin" or "human constant region" as used in the current application denotes a constant heavy chain region of a human antibody of the subclass IgGl, IgG2, IgG3, or IgG4 and/or a constant light chain kappa or lambda region. Such constant regions are well known in the state of the art and e.g.
described by Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991) (see also e.g.
Johnson, G., and Wu, T.T., Nucleic Acids Res. 28 (2000) 214-218; Kabat, E.A., et al., Proc.
Natl. Acad. Sci.
USA 72 (1975) 2785-2788). Within the application for the numbering of positions and mutations the EU numbering system (EU Index) according to Kabat, E.A., et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991) is used and referred to as "numbering according to EU Index of Kabat".
In one embodiment the antagonistic anti-PD1 antibody which binds to human PD1 used in the combination therapy described herein is nivolumab or pembrolizumab and is characterized in comprising the following VH and VL sequences as described herein:
TABLE:
anti-PD-Li antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
nivolumab 1 2 pembrolizumab 3 4 In one preferred embodiment of the invention the compound of formula I used in the combination therapy described herein is selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[5(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
- 44 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD1 antibody used in the combination therapy is nivolumab or pembrolizumab In one embodiment the antagonistic anti-PD1 antibody which binds to human PD1 used in the combination therapy described herein is either a mono- or multispecific antagonistic PD1 antibody and comprises the following heavy chain variable domain VH and light chain variable domain VL sequences as described herein:
TABLE:
anti-PD1 antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
Preferably such anti-PD1 antibody based on the heavy chain variable domain VH
and light chain variable domain VL sequences of PD1-0103-0312 comprises a heavy chain constant region of IgG1 subtype (e.g. SEQ ID NO: 16 or SEQ ID NO: 17, eventually also comprising further mutations, see below the bispecific embodiment) and a human kappa light chain constant region (e.g. SEQ ID NO: 15).
In one embodiment such anti-PD1 antibody based on the heavy chain variable domain VH
and light chain variable domain VL sequences of PD1-0103-0312 is e.g.
bispecific and i) the bispecific antibody comprises a constant heavy chain region of human IgG1 subclass comprising the mutations L234A, L235A and P329G (numberings according to EU
Index of Kabat); and wherein ii)) in the constant heavy chain region a 5354C and mutations are comprised in one CH3 domain and Y349C, T3665, L368A and Y407V
mutations are comprised the other CH3 domain (numberings according to EU Index of Kabat).
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD1 antibody used in the combination therapy is nivolumab or pembrolizumab In one embodiment the antagonistic anti-PD1 antibody which binds to human PD1 used in the combination therapy described herein is either a mono- or multispecific antagonistic PD1 antibody and comprises the following heavy chain variable domain VH and light chain variable domain VL sequences as described herein:
TABLE:
anti-PD1 antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
Preferably such anti-PD1 antibody based on the heavy chain variable domain VH
and light chain variable domain VL sequences of PD1-0103-0312 comprises a heavy chain constant region of IgG1 subtype (e.g. SEQ ID NO: 16 or SEQ ID NO: 17, eventually also comprising further mutations, see below the bispecific embodiment) and a human kappa light chain constant region (e.g. SEQ ID NO: 15).
In one embodiment such anti-PD1 antibody based on the heavy chain variable domain VH
and light chain variable domain VL sequences of PD1-0103-0312 is e.g.
bispecific and i) the bispecific antibody comprises a constant heavy chain region of human IgG1 subclass comprising the mutations L234A, L235A and P329G (numberings according to EU
Index of Kabat); and wherein ii)) in the constant heavy chain region a 5354C and mutations are comprised in one CH3 domain and Y349C, T3665, L368A and Y407V
mutations are comprised the other CH3 domain (numberings according to EU Index of Kabat).
- 45 -In another preferred embodiment of the invention the compound of formula I
used in the combination therapy described herein is selected from the following:
6-Amino-9- [(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(5)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD1 antibody used in the combination therapy comprises a heavy chain variable domain VH with an amino acid sequence of SEQ ID NO: 5 and a light chain variable domain VL with an amino acid sequence of SEQ ID NO: 6.
In one embodiment the antibody which binds to human PD-Li used in the combination therapy described herein is atezolizumab or durvalumab or avelumab and is characterized in comprising the following VH and VL sequences as described herein:
TABLE:
anti-PD-Li antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
atezolizumab 7 8 durvalumab 9 10 avelumab 11 12
used in the combination therapy described herein is selected from the following:
6-Amino-9- [(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(5)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD1 antibody used in the combination therapy comprises a heavy chain variable domain VH with an amino acid sequence of SEQ ID NO: 5 and a light chain variable domain VL with an amino acid sequence of SEQ ID NO: 6.
In one embodiment the antibody which binds to human PD-Li used in the combination therapy described herein is atezolizumab or durvalumab or avelumab and is characterized in comprising the following VH and VL sequences as described herein:
TABLE:
anti-PD-Li antibody amino acid sequence of the amino acid sequence of the heavy chain variable light chain variable domain domain VH, SEQ ID NO: VL, SEQ ID NO:
atezolizumab 7 8 durvalumab 9 10 avelumab 11 12
- 46 -In another preferred embodiment of the invention the compounds of formula I
used in the combination therapy described herein are selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl] -N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD-Li antibody used in the combination therapy is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab).
Another aspect of the invention is the combined treatment (combination treatment) of a patient suffering from liver cancer with the compound of formula I as described above in combination with an anti-angiogenic agent. The anti-angiogenic agent can be co-administered either with compounds of formula I alone or in addition to the combination therapy of the compounds of formula I with an anti-PD-Ll/PD1 axis treatment.
Antiangiogenic agents as used herein include (but are not limited to) small molecule tyrosine kinase inhibitors (TKIs) that bind competitively to the intracellular receptor domains for VEGF, PDGF, and other angiogenic growth factors, like e.g.
sorafenib (4- {4-[3-(4-Chlor-3-trifluormethylphenyl)ureido]phenoxy}pyridin-2-carbonsauremethylamid;
Nexavarrm), regorafenib (4-[4-({[4-Chlor-3-(trifluormethyl)phenyl]carbamoyl}amino)-3-fluorphenoxy]-N-methylpyridin-2-carboxamid-Hydrat; StivargaTm), and sunitinib (N-[2-(Diethylamino)ethyl] -5- [(Z)-(5-fluor- 1,2- dihydro-2-oxo-3 H-indo1-3 -yliden)-methyl] -2,4-dimethy1-1H-pyrrol-3-carboxamid; Sutenin"), but include also anti-VEGF or anti-VEGF
receptor antibodies like e.g. bevacizumab (Avastinn").
used in the combination therapy described herein are selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl] -N-methyl-8-oxo-purine-7-carboxamide);
and the antagonistic PD-Li antibody used in the combination therapy is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab).
Another aspect of the invention is the combined treatment (combination treatment) of a patient suffering from liver cancer with the compound of formula I as described above in combination with an anti-angiogenic agent. The anti-angiogenic agent can be co-administered either with compounds of formula I alone or in addition to the combination therapy of the compounds of formula I with an anti-PD-Ll/PD1 axis treatment.
Antiangiogenic agents as used herein include (but are not limited to) small molecule tyrosine kinase inhibitors (TKIs) that bind competitively to the intracellular receptor domains for VEGF, PDGF, and other angiogenic growth factors, like e.g.
sorafenib (4- {4-[3-(4-Chlor-3-trifluormethylphenyl)ureido]phenoxy}pyridin-2-carbonsauremethylamid;
Nexavarrm), regorafenib (4-[4-({[4-Chlor-3-(trifluormethyl)phenyl]carbamoyl}amino)-3-fluorphenoxy]-N-methylpyridin-2-carboxamid-Hydrat; StivargaTm), and sunitinib (N-[2-(Diethylamino)ethyl] -5- [(Z)-(5-fluor- 1,2- dihydro-2-oxo-3 H-indo1-3 -yliden)-methyl] -2,4-dimethy1-1H-pyrrol-3-carboxamid; Sutenin"), but include also anti-VEGF or anti-VEGF
receptor antibodies like e.g. bevacizumab (Avastinn").
-47 -In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
- 48 -Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethyl]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl]-N-methyl-carbamate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethyl]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl]-N-methyl-carbamate;
- 49 -2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methyl-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2- [S (R)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(R)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methyl-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2- [S (R)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2- [S(R)-propylsulfonimidoyl] -9-(p-tolylmethyl)purine-7-carboxamide;
- 50 -6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
-51 -6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or
- 52 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl] -N-methyl-8-oxo-purine-7-carboxamide);
and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an antagonistic PD1 or antagonistic PD-Li antibody and an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-pip eridyl)piperidine-l-c arb onyl] -2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-l-c arb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl] -N-methyl-8-oxo-purine-7-carboxamide);
and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an antagonistic PD1 or antagonistic PD-Li antibody and an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7- [4-(1-pip eridyl)piperidine-l-c arb onyl] -2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-l-c arb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 53 -6-Amino-9-benzy1-7-(4-methylpiperazine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 54 -Methyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl] -N-methyl-carbamate;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine-l-carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2- [S (S)-propylsulfonimidoyl] -9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
- 55 -6-Amino-2-[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
- 56 -6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;;
and i) the antagonistic PD1 antibody is nivolumab or pembrolizumab or comprises a heavy chain variable domain VH of SEQ ID NO:5 and a light chain variable domain VL of SEQ ID NO:6;
ii) the antagonistic PD-Li antibody is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;;
and i) the antagonistic PD1 antibody is nivolumab or pembrolizumab or comprises a heavy chain variable domain VH of SEQ ID NO:5 and a light chain variable domain VL of SEQ ID NO:6;
ii) the antagonistic PD-Li antibody is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
- 57 -In one preferred embodiment of the invention the compound of formula I used in the combination therapy with an antagonistic PD1 or antagonistic PD-Li antibody and an anti-angiogenic agent described herein is selected from the following:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and i) the antagonistic PD1 antibody is nivolumab or pembrolizumab or comprises a heavy chain variable domain VH of SEQ ID NO:5 and a light chain variable domain VL of SEQ ID NO:6;
ii) the antagonistic PD-Li antibody is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In the following specific embodiments of the invention are included:
1. A compound of formula (I), N H 2 ....¨R3 N----- N> 0 II..õ.....- ...,,,-----....N
0=S N
1 1 \R2 R (I),
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; or 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; (in one preferred embodiment 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide);
and i) the antagonistic PD1 antibody is nivolumab or pembrolizumab or comprises a heavy chain variable domain VH of SEQ ID NO:5 and a light chain variable domain VL of SEQ ID NO:6;
ii) the antagonistic PD-Li antibody is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) and the anti-angiogenic agent used in the combination therapy is sorafenib, regorafenib, sunitinib or bevacizumab (preferably sorafenib or bevacizumab).
In the following specific embodiments of the invention are included:
1. A compound of formula (I), N H 2 ....¨R3 N----- N> 0 II..õ.....- ...,,,-----....N
0=S N
1 1 \R2 R (I),
- 58 -wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; ( or a pharmaceutical composition or medicament thereof);
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and C1_6alkyl;
R3 is -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyC1_6alkyl, C1_6alkyl, Ci_6alkylcarbonyl(C1_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyC1_6alkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof; ( or a pharmaceutical composition or medicament thereof);
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-dimethylpyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3-fluoro-3-methyl-pyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
and their enantiomers or diastereomers are excluded.
- 59 -2. The compound for use according to embodiment 1, wherein Rl is Ci_6alkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
piperazinyl substituted by C1_6alkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyCi_6alkyl.
3. The compound for use according to embodiment 1 or 2, wherein Rl is ethyl or propyl;
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl.
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
piperazinyl substituted by C1_6alkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is C1_6alkyl or Ci_6alkoxyCi_6alkyl;
R5 is (Ci_6alky1)2NCOOCi_6alkyl, Ci_6alkoxyCi_6alkyl, Ci-6alkoxycarbonyl(Ci_6alkyl)aminoCi_6alkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6alkyl, Ci_6alkoxycarbonylCi_6alkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_6alkylcarbonyl(C1_ 6a1ky1)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyCi_6alkyl.
3. The compound for use according to embodiment 1 or 2, wherein Rl is ethyl or propyl;
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl.
- 60 -4. The compound for use according to embodiment 3, wherein R3 is azetidinyl, 4-methylpiperazinyl, piperidinylpiperidinyl, pyrrolidinyl, acetyl(methyl)aminoethyl(methyl)amino, bis(methoxyethyl)amino, butyl(ethyl)amino, butyl(methyl)amino, butyl(methyl)carbamoyloxyethyl(methyl)amino, diethylcarbamoyloxyethyl(methyl)amino, ethoxycarbonyl(methyl)aminoethyl(methyl)amino, ethoxycarbonylethyl(methyl)amino, ethoxycarbonylisobutyl(methyl)amino, ethoxycarbonylisopentyl(methyl)amino, ethoxycarbonylmethyl(methyl)amino, ethoxycarbonyloxyethyl(methyl)amino, ethoxycarbonyl(phenyl)ethyl(methyl)amino, ethyl(methyl)amino, isobutyl(methyl)amino, isopropoxycarbonylisopentyl(methyl)amino, isopropoxycarbonyl(phenyl)ethyl(methyl)amino, isopropyl(methyl)amino, methoxycarbonyl(methyl)aminoethyl(methyl)amino, methoxyethyl(ethyl)amino, methoxyethyl(methyl)amino, methoxyethyl(propyl)amino, methoxypropyl(methyl)amino, propyl(ethyl)amino, propyl(methyl)amino, propyl(methyl)carbamoyloxyethyl(methyl)amino, pyrrolidinylcarbamoyloxyethyl(methyl)amino, tert-butoxycarbonyl(methyl)aminoethyl(methyl)amino, tert-butoxycarbonylethyl(methyl)amino, tert-butoxycarbonylisopentyl(methyl)amino or tert-butoxycarbonyl(phenyl)ethyl(methyl)amino.
5. The compound for use according to any one of embodiments 1 to 4, wherein Rl is ethyl.
6. The compound for use according to embodiment 1 or 2, wherein R2 is benzyl substituted by halogen or C1_6alkyl.
7. The compound for use according to any one of embodiments 2 to 6, wherein R2 is bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl.
8. The compound for use according to embodiment 7, wherein R2 is bromobenzyl, chlorobenzyl or fluorobenzyl.
9. The compound for use according to embodiment 1 or 2, wherein R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is Ci_6alkyl.
5. The compound for use according to any one of embodiments 1 to 4, wherein Rl is ethyl.
6. The compound for use according to embodiment 1 or 2, wherein R2 is benzyl substituted by halogen or C1_6alkyl.
7. The compound for use according to any one of embodiments 2 to 6, wherein R2 is bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl.
8. The compound for use according to embodiment 7, wherein R2 is bromobenzyl, chlorobenzyl or fluorobenzyl.
9. The compound for use according to embodiment 1 or 2, wherein R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is Ci_6alkyl.
-61 -10. The compound for use according to embodiment 9, wherein R3 is propyl(methyl)amino or ethyl(methyl)amino.
11. The compound for use according to any one of embodiments 1, 2, 6 and 9, wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being substituted by halogen or C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is C1_6alkyl.
12. The compound for use according to embodiment 11, wherein Rl is ethyl;
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino.
13. A compound for use in the treatment or prophylaxis of liver cancer selected from:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
11. The compound for use according to any one of embodiments 1, 2, 6 and 9, wherein Rl is C1_6alkyl;
R2 is benzyl, said benzyl being substituted by halogen or C1_6alkyl;
R3 is -NR4R5, wherein R4 is Ci_6alkyl, R5 is C1_6alkyl.
12. The compound for use according to embodiment 11, wherein Rl is ethyl;
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino.
13. A compound for use in the treatment or prophylaxis of liver cancer selected from:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 62 -6-Amino-9-benzy1-7-(4-methylpiperazine-l-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2-[Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
- 63 -Methyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl] -N-methyl-carbamate;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine- 1 -carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine- 1-carbonyl)purin-8-one;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl] -N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-butyl-N-methyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl pyrrolidine- 1 -carboxylate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N-methyl-N-propyl-carbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] ethyl N,N-diethylcarbamate;
2- [ [6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino]ethyl ethyl carbonate;
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-butyl-9- [(4-chlorophenyl)methyl] -N-methyl-8-oxo-2- [S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine- 1-carbonyl)purin-8-one;
- 64 -6-Amino-2-[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
- 65 -6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
14. The compound according to embodiment 13, selected from:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyOmethy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
14. The compound according to embodiment 13, selected from:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
- 66 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
14. The compound for use according to embodiment 13, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide.
16. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of embodiments 1 to 15, wherein the liver cancer is hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer.
17. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of embodiments 1 to 15, wherein the liver cancer is hepatocellular carcinoma.
18. A pharmaceutical composition or medicament comprising a compound according to any one of embodiments 1 to 15 and a therapeutically inert carrier, for use in the treatment or prophylaxis of liver cancer.
19. The use of a compound according to any one of embodiments 1 to 14 for the preparation of a medicament for the treatment or prophylaxis of liver cancer.
20. A method for the treatment or prophylaxis of liver cancer., which method comprises administering a therapeutically effective amount of a compound as defined in any one of embodiments 1 to 15.
21. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
14. The compound for use according to embodiment 13, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide.
16. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of embodiments 1 to 15, wherein the liver cancer is hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer.
17. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of embodiments 1 to 15, wherein the liver cancer is hepatocellular carcinoma.
18. A pharmaceutical composition or medicament comprising a compound according to any one of embodiments 1 to 15 and a therapeutically inert carrier, for use in the treatment or prophylaxis of liver cancer.
19. The use of a compound according to any one of embodiments 1 to 14 for the preparation of a medicament for the treatment or prophylaxis of liver cancer.
20. A method for the treatment or prophylaxis of liver cancer., which method comprises administering a therapeutically effective amount of a compound as defined in any one of embodiments 1 to 15.
21. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in
- 67 -a) the treatment or prophylaxis of liver cancer in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody, or b) the treatment of a patient suffering from liver cancer in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody.
22. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody.
23. Use of a compound as defined in any one of embodiments 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody.
24. The compound, composition, medicament or use, according to any one of embodiments 21 to 23, wherein the treatment is in combination with an antagonistic PD1 antibody.
25. The compound, composition, medicament or use, according to embodiment 24, wherein the antagonistic PD1 antibody is nivolumab or pemprolizumab.
26. The compound, composition, medicament or use, according to embodiment 24, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide 27. The compound, composition, medicament or use, according to embodiment 23, wherein the antagonistic PD1 antibody comprises a heavy chain variable domain VH with an amino acid sequence of SEQ ID NO: 5 and a light chain variable domain VL
with an amino acid sequence of SEQ ID NO:6.
22. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody.
23. Use of a compound as defined in any one of embodiments 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-Li antibody.
24. The compound, composition, medicament or use, according to any one of embodiments 21 to 23, wherein the treatment is in combination with an antagonistic PD1 antibody.
25. The compound, composition, medicament or use, according to embodiment 24, wherein the antagonistic PD1 antibody is nivolumab or pemprolizumab.
26. The compound, composition, medicament or use, according to embodiment 24, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide 27. The compound, composition, medicament or use, according to embodiment 23, wherein the antagonistic PD1 antibody comprises a heavy chain variable domain VH with an amino acid sequence of SEQ ID NO: 5 and a light chain variable domain VL
with an amino acid sequence of SEQ ID NO:6.
- 68 -28. The compound, composition, medicament or use, according to embodiment 27, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-purine-7-carboxamide.
29. The compound, composition, medicament or use, according to any one of embodiments 21 to 23, wherein the treatment is in combination with an antagonistic PD-Li antibody.
30. The compound, composition, medicament or use, according to embodiment 29, wherein the antagonistic PD-Li antibody used in the combination therapy is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) 31. The compound, composition, medicament or use, according to embodiment 30, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide.
32. The compound, composition, medicament or use according to any one of embodiments 21 to 31 wherein additionally an anti-angiogenic agent is used in the combination therapy 33. The compound, composition, medicament or use according to any one of embodiments 21 to 31 wherein additionally an anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab) is used in the combination therapy.
34. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in a) the treatment or prophylaxis of liver cancer in combination with an anti-angiogenic agent, or b) the treatment of a patient suffering from liver cancer in combination with an anti-angiogenic agent.
29. The compound, composition, medicament or use, according to any one of embodiments 21 to 23, wherein the treatment is in combination with an antagonistic PD-Li antibody.
30. The compound, composition, medicament or use, according to embodiment 29, wherein the antagonistic PD-Li antibody used in the combination therapy is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab) 31. The compound, composition, medicament or use, according to embodiment 30, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide.
32. The compound, composition, medicament or use according to any one of embodiments 21 to 31 wherein additionally an anti-angiogenic agent is used in the combination therapy 33. The compound, composition, medicament or use according to any one of embodiments 21 to 31 wherein additionally an anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab) is used in the combination therapy.
34. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in a) the treatment or prophylaxis of liver cancer in combination with an anti-angiogenic agent, or b) the treatment of a patient suffering from liver cancer in combination with an anti-angiogenic agent.
- 69 -35. A compound as defined in any one of embodiments 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
36. Use of a compound as defined in any one of embodiments 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
37. The compound, composition, medicament or use according to any one of embodiments 34 to 36 wherein the anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab).
38. The compound, composition, medicament or use, according to embodiment 37, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-purine-7-carboxamide.
39. The invention as hereinbefore described.
Description of the amino acid sequences SEQ ID NO: 1 heavy chain variable domain of anti-PD1 antibody nivolumab SEQ ID NO: 2 light chain variable domain of anti-PD1 antibody nivolumab SEQ ID NO: 3 heavy chain variable domain of anti-PD1 antibody pembrolizumab SEQ ID NO: 4 light chain variable domain of anti-PD1 antibody pembrolizumab SEQ ID NO: 5 heavy chain variable domain of anti-PD1 antibody PD1-SEQ ID NO: 6 light chain variable domain of anti-PD1 antibody PD1-SEQ ID NO: 7 heavy chain variable domain of anti-PD-Li antibody atezolizumab SEQ ID NO: 8 light chain variable domain of anti-PD-Li antibody atezolizumab SEQ ID NO: 9 heavy chain variable domain of anti-PD-Li antibody durvalumab SEQ ID NO: 10 light chain variable domain of anti-PD-Li antibody durvalumab
36. Use of a compound as defined in any one of embodiments 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
37. The compound, composition, medicament or use according to any one of embodiments 34 to 36 wherein the anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab).
38. The compound, composition, medicament or use, according to embodiment 37, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-purine-7-carboxamide.
39. The invention as hereinbefore described.
Description of the amino acid sequences SEQ ID NO: 1 heavy chain variable domain of anti-PD1 antibody nivolumab SEQ ID NO: 2 light chain variable domain of anti-PD1 antibody nivolumab SEQ ID NO: 3 heavy chain variable domain of anti-PD1 antibody pembrolizumab SEQ ID NO: 4 light chain variable domain of anti-PD1 antibody pembrolizumab SEQ ID NO: 5 heavy chain variable domain of anti-PD1 antibody PD1-SEQ ID NO: 6 light chain variable domain of anti-PD1 antibody PD1-SEQ ID NO: 7 heavy chain variable domain of anti-PD-Li antibody atezolizumab SEQ ID NO: 8 light chain variable domain of anti-PD-Li antibody atezolizumab SEQ ID NO: 9 heavy chain variable domain of anti-PD-Li antibody durvalumab SEQ ID NO: 10 light chain variable domain of anti-PD-Li antibody durvalumab
- 70 -SEQ ID NO: 11 heavy chain variable domain of anti-PD-Li antibody avelumab SEQ ID NO: 12 light chain variable domain of anti-PD-Li antibody avelumab SEQ ID NO: 13 exemplary human PD-Li SEQ ID NO: 14 exemplary human PD1 SEQ ID NO: 15 human kappa light chain constant region SEQ ID NO: 16 human heavy chain constant region derived from IgG1 SEQ ID NO: 17 human heavy chain constant region derived from IgG1 mutated on L234A, L235A, P329G.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1: Combination of a prodrug form of the compounds of the present invention (compound 41-A) and Sorafenib results in 2 tumor-free mice in the iAST
mouse model of hepatocellular carcinoma Figure 1A: Synergistic effect of Compound 41-A and sorafenib on tumor burden (tumor free mice) Figure 1B: Combined liver and tumor weight after treatment:
Figure 2: Treatment with a prodrug form of the compounds of the present invention (compound 41-A) induces PD-Li expression on tumor cells in the iAST
mouse model of hepatocellular carcinoma. Figure 2A: CD45+ total immune cell infiltrate, Figure 2B: PD-Li on CD45-, Figure 2C: CD1 lb-lymphoid cells, Figure 2D: CD1 lb+ myeloid cells Figure 3: Triple combination of with a prodrug form of the compounds of the present invention (compound 41-A), Sorafenib and anti-PD-1 results in increased median survival.
Figure 4: Treatment with a prodrug form of the compounds of the present invention (compound 41-A) results in tumor stasis in the transplanted Hep55.1c mouse model of hepatocellular carcinoma.
Figure 5A: Combination of a prodrug form of the compounds of the present invention (compound 41-A) and anti-PD-1 antibodies results in survival benefit in the Hep55.1c mouse model of hepatocellular carcinoma.
Figure 5B: In vivo efficacy of compound 42-A (6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[5(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-purine-7-carboxamide), alone and in combination with anti-PD-1 in hepatocellular carcinoma.
Figure 6: Treatment with an active form of the compounds of the present invention does not induce enhanced tumor cell proliferation in cell lines originating
BRIEF DESCRIPTION OF THE FIGURES
Figure 1: Combination of a prodrug form of the compounds of the present invention (compound 41-A) and Sorafenib results in 2 tumor-free mice in the iAST
mouse model of hepatocellular carcinoma Figure 1A: Synergistic effect of Compound 41-A and sorafenib on tumor burden (tumor free mice) Figure 1B: Combined liver and tumor weight after treatment:
Figure 2: Treatment with a prodrug form of the compounds of the present invention (compound 41-A) induces PD-Li expression on tumor cells in the iAST
mouse model of hepatocellular carcinoma. Figure 2A: CD45+ total immune cell infiltrate, Figure 2B: PD-Li on CD45-, Figure 2C: CD1 lb-lymphoid cells, Figure 2D: CD1 lb+ myeloid cells Figure 3: Triple combination of with a prodrug form of the compounds of the present invention (compound 41-A), Sorafenib and anti-PD-1 results in increased median survival.
Figure 4: Treatment with a prodrug form of the compounds of the present invention (compound 41-A) results in tumor stasis in the transplanted Hep55.1c mouse model of hepatocellular carcinoma.
Figure 5A: Combination of a prodrug form of the compounds of the present invention (compound 41-A) and anti-PD-1 antibodies results in survival benefit in the Hep55.1c mouse model of hepatocellular carcinoma.
Figure 5B: In vivo efficacy of compound 42-A (6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[5(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-purine-7-carboxamide), alone and in combination with anti-PD-1 in hepatocellular carcinoma.
Figure 6: Treatment with an active form of the compounds of the present invention does not induce enhanced tumor cell proliferation in cell lines originating
- 71 -from hepatocellular carcinoma and cholangiocarcinoma.Figure 6A:
compound 41c-B, Figure 6B: compound 41c-A
Figure 7: 7A and 7B: Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-B) result in inhibition of proliferation in tumor cell lines. Figure 7A: Cell lines Hep3B, SNU449, HLF, JHH2, Huh7, OZ, JHH1, HepG2, Figure 7B:
Cell lines JHH4, HLE, JHH6, JHH5, SIcHepl, EGI 1 .
7C: Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-A) result in inhibition of proliferation in tumor cell lines.
Figure 8: Single crystal X-ray diffraction of Example 41-B.
Figure 9: Single crystal X-ray diffraction of Example 42-A.
Figure 10: Single crystal X-ray diffraction of Example 43-B.
EXAMPLES
The invention will be more fully understood by reference to the following examples.
They should not, however, be construed as limiting the scope of the invention.
ABBREVIATIONS
aq. aqueous BSA: N, 0-bis(trimethylsilyl)acetamide CDI: /V,N'-carbonyl diimidazole DIEPA: N, N-diethylpropylamine DBU: 1,8-Diazabicycloundec-7-ene DPPA: diphenylphosphoryl azide ECso: the molar concentration of an agonist, which produces 50% of the maximum possible response for that agonist.
EDC: N1-((ethylimino)methylene)-N3,/V3-dimethylpropane-1,3-diamine Et0Ac or EA: ethyl acetate
compound 41c-B, Figure 6B: compound 41c-A
Figure 7: 7A and 7B: Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-B) result in inhibition of proliferation in tumor cell lines. Figure 7A: Cell lines Hep3B, SNU449, HLF, JHH2, Huh7, OZ, JHH1, HepG2, Figure 7B:
Cell lines JHH4, HLE, JHH6, JHH5, SIcHepl, EGI 1 .
7C: Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-A) result in inhibition of proliferation in tumor cell lines.
Figure 8: Single crystal X-ray diffraction of Example 41-B.
Figure 9: Single crystal X-ray diffraction of Example 42-A.
Figure 10: Single crystal X-ray diffraction of Example 43-B.
EXAMPLES
The invention will be more fully understood by reference to the following examples.
They should not, however, be construed as limiting the scope of the invention.
ABBREVIATIONS
aq. aqueous BSA: N, 0-bis(trimethylsilyl)acetamide CDI: /V,N'-carbonyl diimidazole DIEPA: N, N-diethylpropylamine DBU: 1,8-Diazabicycloundec-7-ene DPPA: diphenylphosphoryl azide ECso: the molar concentration of an agonist, which produces 50% of the maximum possible response for that agonist.
EDC: N1-((ethylimino)methylene)-N3,/V3-dimethylpropane-1,3-diamine Et0Ac or EA: ethyl acetate
- 72 -HATU: (1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate) hr(s): hour(s) HPLC: high performance liquid chromatography HOBt: N-hydroxybenzotriazole MS (ESI): mass spectroscopy (electron spray ionization) m-CPBA: 3-chloroperbenzoic acid MTEB: methyl tert-butyl ether NMP: N-methylpyrrolidone obsd. observed PE: petroleum ether PMB: p-methoxybenzyl PPA: polyphosphoric acid QOD every other day QW once a week RT or rt: room temperature sat. saturated TFA: trifluoroacetic acid TEA: triethylamine V/V volume ratio GENERAL EXPERIMENTAL CONDITIONS
Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module. ii) ISCO combi-flash chromatography instrument. Silica gel Brand and pore size: i)
Intermediates and final compounds were purified by flash chromatography using one of the following instruments: i) Biotage SP1 system and the Quad 12/25 Cartridge module. ii) ISCO combi-flash chromatography instrument. Silica gel Brand and pore size: i)
-73 -KP-SIL 60 A, particle size: 40-60 lam; ii) CAS registry NO: Silica Gel: 63231-67-4, particle size: 47-60 micron silica gel; iii) ZCX from Qingdao Haiyang Chemical Co., Ltd, pore: 200-300 or 300-400.
Intermediates and final compounds were purified by preparative HPLC on reversed phase column using X BridgeTM Perp C18 (5 lam, OBDTM 30 x 100 mm) column or SunFireTM Perp C18 (5 lam, OBDTM 30 x 100 mm) column.
LC/MS spectra were obtained using a Waters UPLC-SQD Mass. Standard LC/MS
conditions were as follows (running time 3 minutes):
Acidic condition: A: 0.1% formic acid and 1% acetonitrile in H20; B: 0.1%
formic acid in acetonitrile;
Basic condition: A: 0.05% NI-13.H20 in H20; B: acetonitrile.
Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (M+H) .
NMR Spectra were obtained using Bruker Avance 400MHz.
All reactions involving air-sensitive reagents were performed under an argon atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.
PREPARATIVE EXAMPLES
Preparation of Intermediate Intermediate AA
N-methyl-N-propyl-carbamoyl chloride CI N
I
AA
Intermediates and final compounds were purified by preparative HPLC on reversed phase column using X BridgeTM Perp C18 (5 lam, OBDTM 30 x 100 mm) column or SunFireTM Perp C18 (5 lam, OBDTM 30 x 100 mm) column.
LC/MS spectra were obtained using a Waters UPLC-SQD Mass. Standard LC/MS
conditions were as follows (running time 3 minutes):
Acidic condition: A: 0.1% formic acid and 1% acetonitrile in H20; B: 0.1%
formic acid in acetonitrile;
Basic condition: A: 0.05% NI-13.H20 in H20; B: acetonitrile.
Mass spectra (MS): generally only ions which indicate the parent mass are reported, and unless otherwise stated the mass ion quoted is the positive mass ion (M+H) .
NMR Spectra were obtained using Bruker Avance 400MHz.
All reactions involving air-sensitive reagents were performed under an argon atmosphere. Reagents were used as received from commercial suppliers without further purification unless otherwise noted.
PREPARATIVE EXAMPLES
Preparation of Intermediate Intermediate AA
N-methyl-N-propyl-carbamoyl chloride CI N
I
AA
- 74 -To a mixture of N-methylpropan- 1 -amine (5 g, 68.4 mmol) and sodium hydrogencarbonate (11.5 g, 137 mmol) in DCM (70 mL) at 0 C was added bis(trichloromethyl) carbonate (8.11 g, 27.3 mmol) in DCM (30 mL) dropwise.
The mixture was stirred at room temperature for 2 hrs and filtered. The filtrate was concentrated in vacuo. The obtained N-methyl-N-propyl-carbamoyl chloride (7.2 g, Intermediate AA) was used for next step without further purification.
Intermediate AB
N-(2-Methoxyethyl)-N-methyl-carbamoyl chloride a N
I
AB
Intermediate AB was prepared in analogy to Intermediate AA by using 2-methoxy-N-methyl-ethanamine instead of N-methylpropan-l-amine. N-(2-Methoxyethyl)-N-methyl-carbamoyl chloride (8 g, Intermediate AB) was obtained and used for next step without further purification.
Intermediate AC
N-Ethyl-N-propyl-carbamoyl chloride /--/
N
CI \_ AC
Intermediate AC was prepared in analogy to Intermediate AA by using N-ethylpropan-l-amine instead of N-methylpropan-l-amine. N-Ethyl-N-propyl-carbamoyl chloride (12.6 g, Intermediate AC) was obtained as a yellow oil and used for next step without further purification.
The mixture was stirred at room temperature for 2 hrs and filtered. The filtrate was concentrated in vacuo. The obtained N-methyl-N-propyl-carbamoyl chloride (7.2 g, Intermediate AA) was used for next step without further purification.
Intermediate AB
N-(2-Methoxyethyl)-N-methyl-carbamoyl chloride a N
I
AB
Intermediate AB was prepared in analogy to Intermediate AA by using 2-methoxy-N-methyl-ethanamine instead of N-methylpropan-l-amine. N-(2-Methoxyethyl)-N-methyl-carbamoyl chloride (8 g, Intermediate AB) was obtained and used for next step without further purification.
Intermediate AC
N-Ethyl-N-propyl-carbamoyl chloride /--/
N
CI \_ AC
Intermediate AC was prepared in analogy to Intermediate AA by using N-ethylpropan-l-amine instead of N-methylpropan-l-amine. N-Ethyl-N-propyl-carbamoyl chloride (12.6 g, Intermediate AC) was obtained as a yellow oil and used for next step without further purification.
- 75 -Intermediate AD
N-Ethyl-N-(2-methoxyethyl)carbamoyl chloride N
CI ) AD
Intermediate AD was prepared in analogy to Intermediate AA by using N-ethyl-2-methoxyethanamine instead of N-methylpropan-l-amine. The crude N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (2.5 g, Intermediate AD) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AE
N-Butyl-N-ethyl-carbamoyl chloride N
Intermediate AE was prepared in analogy to Intermediate AA by using N-ethylbutan- 1 -amine (5 g) instead of N-methylpropan- 1 -amine. The crude N-butyl-N-ethyl-carbamoyl chloride (6.3 g, Intermediate AE) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AF
N-(2-Methoxyethyl)-N-propyl-carbamoyl chloride 0¨
0 /¨/
N
CI
AF
N-Ethyl-N-(2-methoxyethyl)carbamoyl chloride N
CI ) AD
Intermediate AD was prepared in analogy to Intermediate AA by using N-ethyl-2-methoxyethanamine instead of N-methylpropan-l-amine. The crude N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (2.5 g, Intermediate AD) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AE
N-Butyl-N-ethyl-carbamoyl chloride N
Intermediate AE was prepared in analogy to Intermediate AA by using N-ethylbutan- 1 -amine (5 g) instead of N-methylpropan- 1 -amine. The crude N-butyl-N-ethyl-carbamoyl chloride (6.3 g, Intermediate AE) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AF
N-(2-Methoxyethyl)-N-propyl-carbamoyl chloride 0¨
0 /¨/
N
CI
AF
- 76 -Intermediate AF was prepared in analogy to Intermediate AA by using N-(2-methoxyethyl)propan-1-amine (2 g, 17.1 mmol) instead of N-methylpropan-l-amine. The crude N-(2-methoxyethyl)-N-propyl-carbamoyl chloride (2.5 g, Intermediate AF) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AG
N,N-Bis(2-methoxyethyl)carbamoyl chloride \
N
CI
\--\
0¨
AG
Intermediate AG was prepared in analogy to Intermediate AA by using of bis(2-methoxyethyl)amine (2 g, 15 mmol) instead of N-methylpropan- 1 -amine. The crude product /V,N-bis(2-methoxyethyl)carbamoyl chloride (2.6 g, Intermediate AG) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AH
Azetidine-l-carbonyl chloride CI
AH
Intermediate AH was prepared in analogy to Intermediate AA by using azetidine hydrochloride (10.7 g, 107 mmol) and sodium bicarbonate (3 equiv.) instead of N-methylpropan- 1 -amine and sodium bicarbonate (2 equiv.). The crude azetidine-1 -carbonyl chloride (1.5 g, Intermediate AH) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AG
N,N-Bis(2-methoxyethyl)carbamoyl chloride \
N
CI
\--\
0¨
AG
Intermediate AG was prepared in analogy to Intermediate AA by using of bis(2-methoxyethyl)amine (2 g, 15 mmol) instead of N-methylpropan- 1 -amine. The crude product /V,N-bis(2-methoxyethyl)carbamoyl chloride (2.6 g, Intermediate AG) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AH
Azetidine-l-carbonyl chloride CI
AH
Intermediate AH was prepared in analogy to Intermediate AA by using azetidine hydrochloride (10.7 g, 107 mmol) and sodium bicarbonate (3 equiv.) instead of N-methylpropan- 1 -amine and sodium bicarbonate (2 equiv.). The crude azetidine-1 -carbonyl chloride (1.5 g, Intermediate AH) was obtained as a light yellow oil and used for next step without further purification.
- 77 -Intermediate Al N-Isopropyl-N-methyl-carbamoyl chloride N
CI \
Al Intermediate Al was prepared in analogy to Intermediate AA by using N-methylpropan-2-amine (5 g, 19.4 mmol) instead of N-methylpropan- 1-amine. The crude N-isopropyl-N-methyl-carbamoyl chloride (8.6 g, Intermediate Al) was obtained as a yellow oil and used for next step without further purification.
Intermediate AL
N-Isobutyl-N-methyl-carbamoyl chloride CI \
AL
Intermediate AL was prepared in analogy to Intermediate AA by using N-2-dimethylpropan-1-amine (4.8 g) instead of N-methylpropan-l-amine. The crude N-isobutyl-N-methyl-carbamoyl chloride (8.1 g, Intermediate AL) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AP
Ethyl 2-Ichlorocarbonyhmethyl)amino]acetate 0¨/
0 ______________________________________ / µ
,¨N 0 CI \
AP
CI \
Al Intermediate Al was prepared in analogy to Intermediate AA by using N-methylpropan-2-amine (5 g, 19.4 mmol) instead of N-methylpropan- 1-amine. The crude N-isopropyl-N-methyl-carbamoyl chloride (8.6 g, Intermediate Al) was obtained as a yellow oil and used for next step without further purification.
Intermediate AL
N-Isobutyl-N-methyl-carbamoyl chloride CI \
AL
Intermediate AL was prepared in analogy to Intermediate AA by using N-2-dimethylpropan-1-amine (4.8 g) instead of N-methylpropan-l-amine. The crude N-isobutyl-N-methyl-carbamoyl chloride (8.1 g, Intermediate AL) was obtained as a light yellow oil and used for next step without further purification.
Intermediate AP
Ethyl 2-Ichlorocarbonyhmethyl)amino]acetate 0¨/
0 ______________________________________ / µ
,¨N 0 CI \
AP
- 78 -To a solution of triphosgene (728 mg, 2.45 mmol) in DCM (5 mL) was added a solution of ethyl 2-(methylamino)acetate hydrochloride (1.3 g, 8.46 mmol) and pyridine (1 mL) in DCM (5 mL) dropwise at 0 C. The reaction mixture became orange and a yellow precipitate appeared, then it was allowed to warm to room temperature. After stirred for 1 hr, aqueous HC1 (0.1N, 25 mL) was added to the reaction mixture, the organic layer was separated, washed with 0.1 N HC1 (10 mL) twice, brine (10 mL), dried over Na2SO4 and concentrated in vacuo to give the crude ethyl 2-[chlorocarbonyl(methyl)amino]acetate (2.0 g, Intermediate AP) as a light yellow oil and used for next step without further purification.
Intermediate AR
tert-Butyl 3-Ich1orocarbony1(methyl)amino]propanoate 0 y 0 ,_0 _11/
CI \
AR
Step 1: Preparation of tert-butyl 3-(methylamino)propanoate (Compound AR-1) 0 y H N, \
To a solution of tert-butyl acrylate (3 g) in DMF (40 mL) was added methylamine hydrochloride (4.74 g, 70 mmol) and DBU (21.4 g, 140 mmol) at -45 C. Then the reaction temperature was allowed to warm to -10 C. The reaction mixture was stirred at the same temperature for 2.5 hrs. Et20 (200 mL) was added and the resulting mixture was washed with brine (50 mL) four times. The separated organic layer was dried over Na2SO4 and concentrated in vacuo to afford tert-butyl 3-(methylamino)propanoate (3.5 g, Compound AR-1) as a light yellow oil.
Step 2: Preparation of tert-butyl 3-Ich1orocarbony1(methyl)amino]propanoate
Intermediate AR
tert-Butyl 3-Ich1orocarbony1(methyl)amino]propanoate 0 y 0 ,_0 _11/
CI \
AR
Step 1: Preparation of tert-butyl 3-(methylamino)propanoate (Compound AR-1) 0 y H N, \
To a solution of tert-butyl acrylate (3 g) in DMF (40 mL) was added methylamine hydrochloride (4.74 g, 70 mmol) and DBU (21.4 g, 140 mmol) at -45 C. Then the reaction temperature was allowed to warm to -10 C. The reaction mixture was stirred at the same temperature for 2.5 hrs. Et20 (200 mL) was added and the resulting mixture was washed with brine (50 mL) four times. The separated organic layer was dried over Na2SO4 and concentrated in vacuo to afford tert-butyl 3-(methylamino)propanoate (3.5 g, Compound AR-1) as a light yellow oil.
Step 2: Preparation of tert-butyl 3-Ich1orocarbony1(methyl)amino]propanoate
- 79 -(Intermediate AR) 0 y 0 )_ N
\/
CI µ
AR
Intermediate AR was prepared in analogy to Intermediate AP by using tert-butyl 3-(methylamino)propanoate (3.4 g, Compound AR-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl 3-[chlorocarbonyl(methyl)amino]propanoate (3.5 g, Intermediate AR) was obtained and used for next step without further purification.
Intermediate AS
Ethyl (2S)-2-Ichlorocarbonyhmethyl)amino]propanoate \ 0-/
0 ______________________________________ 1 µ0 N
CI \
AS
Step 1: Preparation of ethyl (2S)-2-(methylamino)propanoate hydrochloride (Compound AS-1) YO
H N HCI
To a solution of (25)-2-(methylamino)propanoic acid (1 g, 9.70 mmol) in Et0H
(10 mL) was added SOC12 (1.50 g, 12.61 mmol) dropwise at 0 C in 0.5 hr. The reaction mixture was stirred at 25 C for 15.5 hrs, then diluted with EA (20 mL), washed with H20 (5 mL) and brine (5 mL). The organic layer was dried over Na2SO4 and concentrated in vacuo. Ethyl (2S)-2-(methylamino)propanoate hydrochloride (1.8 g, Compound AS-1)
\/
CI µ
AR
Intermediate AR was prepared in analogy to Intermediate AP by using tert-butyl 3-(methylamino)propanoate (3.4 g, Compound AR-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl 3-[chlorocarbonyl(methyl)amino]propanoate (3.5 g, Intermediate AR) was obtained and used for next step without further purification.
Intermediate AS
Ethyl (2S)-2-Ichlorocarbonyhmethyl)amino]propanoate \ 0-/
0 ______________________________________ 1 µ0 N
CI \
AS
Step 1: Preparation of ethyl (2S)-2-(methylamino)propanoate hydrochloride (Compound AS-1) YO
H N HCI
To a solution of (25)-2-(methylamino)propanoic acid (1 g, 9.70 mmol) in Et0H
(10 mL) was added SOC12 (1.50 g, 12.61 mmol) dropwise at 0 C in 0.5 hr. The reaction mixture was stirred at 25 C for 15.5 hrs, then diluted with EA (20 mL), washed with H20 (5 mL) and brine (5 mL). The organic layer was dried over Na2SO4 and concentrated in vacuo. Ethyl (2S)-2-(methylamino)propanoate hydrochloride (1.8 g, Compound AS-1)
- 80 -was obtained as a yellow oil and used for next step without further purification.
Step 2: Preparation of ethyl (2S)-2-(methylamino)propanoate (Compound AS-2) YO
H N
A solution of ethyl (2S)-2-(methylamino)propanoate hydrochloride (1.8 g, Compound AS-1) in EA (10 mL) was adjusted to pH = 8 with 10 wt. % aqueous NaHCO3.
The reaction mixture was stirred at room temperature for 0.5 hr. The organic layer was washed with brine (5 mL), dried over Na2SO4 and concentrated in vacuo. Ethyl (2S)-2-(methylamino)propanoate (620 mg, Compound AS-2) was obtained as a yellow oil and used for the next step without further purification.
Step 3: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]propanoate (Intermediate AS) µ 0-/
CI \
AS
Intermediate AS was prepared in analogy to Intermediate AP by using ethyl (2S)-2-(methylamino)propanoate (260 mg, Compound AS-2) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (25)-2-[chlorocarbonyl(methyl)amino]propanoate (200 mg, Intermediate AS) was obtained as a yellow oil and used for the next step without further purification.
Intermediate AT
tert-Butyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate
Step 2: Preparation of ethyl (2S)-2-(methylamino)propanoate (Compound AS-2) YO
H N
A solution of ethyl (2S)-2-(methylamino)propanoate hydrochloride (1.8 g, Compound AS-1) in EA (10 mL) was adjusted to pH = 8 with 10 wt. % aqueous NaHCO3.
The reaction mixture was stirred at room temperature for 0.5 hr. The organic layer was washed with brine (5 mL), dried over Na2SO4 and concentrated in vacuo. Ethyl (2S)-2-(methylamino)propanoate (620 mg, Compound AS-2) was obtained as a yellow oil and used for the next step without further purification.
Step 3: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]propanoate (Intermediate AS) µ 0-/
CI \
AS
Intermediate AS was prepared in analogy to Intermediate AP by using ethyl (2S)-2-(methylamino)propanoate (260 mg, Compound AS-2) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (25)-2-[chlorocarbonyl(methyl)amino]propanoate (200 mg, Intermediate AS) was obtained as a yellow oil and used for the next step without further purification.
Intermediate AT
tert-Butyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate
- 81 -CI \
AT
Step 1: Preparation of tert-butyl (2S)-4-methyl-2-(methylamino)pentanoate (Compound AT-1) e( \
2-Methylpropene (25 g, 446 mmol) was bubbled into DCM (50 mL) at -78 C. Then the 2-methylpropene solution was added to a solution of (5)-4-methy1-2-(methylamino)pentanoic acid hydrochloride (500 mg, 2.75 mmol) and H2SO4 (3.68 g, 2 mL, 37.5 mmol) in dioxane (20 mL) at 0 C. The reaction mixture was stirred at room temperature for 18 hrs in a sealed tube. The reaction solution was poured into an ice cold aqueous KOH solution (8.4 g in water (30mL)) and the resulting mixture was extracted with DCM (50 mL) twice. The combined organic layer was washed with brine (30 mL) twice, dried over Na2SO4 and concentrated in vacuo to afford the crude product tert-butyl (2S)-4-methyl-2-(methylamino)pentanoate (Compound AT-1) as a light yellow oil.
Step 2: Preparation of tert-butyl (2S)-2-Ich1orocarbony1(methyl)amino]-4-methyl-pentanoate (Intermediate AT) 0 <C)( N \ 0 CI \
AT
Intermediate AT was prepared in analogy to Intermediate AP by using tert-butyl
AT
Step 1: Preparation of tert-butyl (2S)-4-methyl-2-(methylamino)pentanoate (Compound AT-1) e( \
2-Methylpropene (25 g, 446 mmol) was bubbled into DCM (50 mL) at -78 C. Then the 2-methylpropene solution was added to a solution of (5)-4-methy1-2-(methylamino)pentanoic acid hydrochloride (500 mg, 2.75 mmol) and H2SO4 (3.68 g, 2 mL, 37.5 mmol) in dioxane (20 mL) at 0 C. The reaction mixture was stirred at room temperature for 18 hrs in a sealed tube. The reaction solution was poured into an ice cold aqueous KOH solution (8.4 g in water (30mL)) and the resulting mixture was extracted with DCM (50 mL) twice. The combined organic layer was washed with brine (30 mL) twice, dried over Na2SO4 and concentrated in vacuo to afford the crude product tert-butyl (2S)-4-methyl-2-(methylamino)pentanoate (Compound AT-1) as a light yellow oil.
Step 2: Preparation of tert-butyl (2S)-2-Ich1orocarbony1(methyl)amino]-4-methyl-pentanoate (Intermediate AT) 0 <C)( N \ 0 CI \
AT
Intermediate AT was prepared in analogy to Intermediate AP by using tert-butyl
- 82 -(2S)-4-methyl-2-(methylamino)pentanoate (300 mg, Compound AT-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (350 mg, Intermediate AT) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AU
Isopropyl (2S)-2-Ichlorocarbonyhmethyl)amino]-4-methyl-pentanoate N \O
CI \
AU
Step 1: Preparation of isopropyl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride (Compound AU-1) µ0¨( H HCI
To a solution of (5)-4-methy1-2-(methylamino)pentanoic acid hydrochloride (0.5 g) in i-PrOH (7.8 g, 10 mL) was added thionyl chloride (655 mg, 402 [EL) dropwise at room temperature. The resulting mixture was stirred and refluxed for 16 hrs and then concentrated in vacuo. The residue was basified with saturated aqueous NaHCO3 (30 mL) and extracted with DCM (50 mL). The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was salified with HC1/Et0Ac (10 mL, 1 mmol/mL) and concentrated to afford isopropyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (510 mg, Compound AU-1) as a white solid.
Intermediate AU
Isopropyl (2S)-2-Ichlorocarbonyhmethyl)amino]-4-methyl-pentanoate N \O
CI \
AU
Step 1: Preparation of isopropyl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride (Compound AU-1) µ0¨( H HCI
To a solution of (5)-4-methy1-2-(methylamino)pentanoic acid hydrochloride (0.5 g) in i-PrOH (7.8 g, 10 mL) was added thionyl chloride (655 mg, 402 [EL) dropwise at room temperature. The resulting mixture was stirred and refluxed for 16 hrs and then concentrated in vacuo. The residue was basified with saturated aqueous NaHCO3 (30 mL) and extracted with DCM (50 mL). The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was salified with HC1/Et0Ac (10 mL, 1 mmol/mL) and concentrated to afford isopropyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (510 mg, Compound AU-1) as a white solid.
- 83 -Step 2: Preparation of isopropyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate (Intermediate AU) N \ 0 CI \
AU
Intermediate AU was prepared in analogy to Intermediate AP by using isopropyl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride (500 mg, Compound AU-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (650 mg, Intermediate AU) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AV
Ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-3-methyl-butanoate 0¨/
0 ______________________________________ CI \
AV
Step 1: Preparation of ethyl (2S)-3-methyl-2-(methylamino)butanoate hydrochloride (Compound AV-1) _____________________________________ µ0¨\
\
To a solution of (25)-3-methy1-2-(methylamino)butanoic acid (1.0 g, 7.6 mmol) in Et0H (10 mL) was added thionyl chloride (2.45 g, 21 mmol) dropwise at room
AU
Intermediate AU was prepared in analogy to Intermediate AP by using isopropyl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride (500 mg, Compound AU-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (650 mg, Intermediate AU) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AV
Ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-3-methyl-butanoate 0¨/
0 ______________________________________ CI \
AV
Step 1: Preparation of ethyl (2S)-3-methyl-2-(methylamino)butanoate hydrochloride (Compound AV-1) _____________________________________ µ0¨\
\
To a solution of (25)-3-methy1-2-(methylamino)butanoic acid (1.0 g, 7.6 mmol) in Et0H (10 mL) was added thionyl chloride (2.45 g, 21 mmol) dropwise at room
- 84 -temperature. The resulting mixture was stirred and refluxed for 16 hrs and then concentrated in vacuo. The residue was basified with saturated aqueous NaHCO3 (30 mL) and extracted with DCM (50 mL) twice. The combined organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was dissolved in HC1/Et0Ac (10 mL, 1 M) and concentrated to afford ethyl (25)-3-methy1-2-(methylamino)butanoate hydrochloride (1.9 g, Compound AV-1) as a white solid.
Step 2: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-3-methyl-butanoate (Intermediate AV) 0¨/
0 ______________________________________ µ
,-N 0 CI \
AV
Intermediate AV was prepared in analogy to Intermediate AP by using ethyl (2S)-3-methy1-2-(methylamino)butanoate hydrochloride (500 mg, Compound AV-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (25)-2-[chlorocarbonyl(methyl)amino]-3-methyl-butanoate (600 mg, Intermediate AV) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AW
Ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate 0¨/
0 ______________________________________ ,-N 0 CI \
AW
Step 2: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-3-methyl-butanoate (Intermediate AV) 0¨/
0 ______________________________________ µ
,-N 0 CI \
AV
Intermediate AV was prepared in analogy to Intermediate AP by using ethyl (2S)-3-methy1-2-(methylamino)butanoate hydrochloride (500 mg, Compound AV-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (25)-2-[chlorocarbonyl(methyl)amino]-3-methyl-butanoate (600 mg, Intermediate AV) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AW
Ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate 0¨/
0 ______________________________________ ,-N 0 CI \
AW
- 85 -Step 1: Preparation of ethyl (2S)-4-methyl-2-(methylamino)pentanoate hydrochloride (Compound AW-1) 0¨/
µ HCI
\
To a solution of (2S)-4-methyl-2-(methylamino)pentanoic acid (1 g, 6.9 mmol) in Et0H (10 mL) was added thionyl chloride (1.07 g, 8.3 mmol) dropwise at room temperature. The resulting mixture was stirred at reflux for 16 hrs and then concentrated in vacuo. The residue was basified with saturated aqueous NaHCO3 (30 mL) and extracted with DCM (50 mL). The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was salified with HCl/Et0Ac (10 mL, lmmol/mL) and concentrated to give ethyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (1.8 g, Compound AW-1) as a white solid.
Step 2: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate (Intermediate AW) 0¨/
0 ______________________________________ ,-N 0 CI \
AW
Intermediate AW was prepared in analogy to Intermediate AP by using ethyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (610 mg, AW-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (280 mg, Intermediate AW) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AX
µ HCI
\
To a solution of (2S)-4-methyl-2-(methylamino)pentanoic acid (1 g, 6.9 mmol) in Et0H (10 mL) was added thionyl chloride (1.07 g, 8.3 mmol) dropwise at room temperature. The resulting mixture was stirred at reflux for 16 hrs and then concentrated in vacuo. The residue was basified with saturated aqueous NaHCO3 (30 mL) and extracted with DCM (50 mL). The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo. The residue was salified with HCl/Et0Ac (10 mL, lmmol/mL) and concentrated to give ethyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (1.8 g, Compound AW-1) as a white solid.
Step 2: Preparation of ethyl (2S)-2-Ichlorocarbonyhmethyllamino]-4-methyl-pentanoate (Intermediate AW) 0¨/
0 ______________________________________ ,-N 0 CI \
AW
Intermediate AW was prepared in analogy to Intermediate AP by using ethyl (25)-4-methy1-2-(methylamino)pentanoate hydrochloride (610 mg, AW-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (280 mg, Intermediate AW) was obtained as a light yellow oil and used for the next step without further purification.
Intermediate AX
- 86 -Ethyl (2S)-2-Ichlorocarbonyl(methyl)amino]-3-phenyl-propanoate Chiral 0¨/
CI \
AX
Intermediate AX was prepared in analogy to Intermediate AP by using (S)-ethy1-2-(methylamino)-3-phenylpropanoate instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (200 mg, Intermediate AX) was obtained as a light yellow oil and used for the next step without further purification Intermediate AY
Isopropyl (2S)-2-Ichlorocarbonyl(methyl)amino]-3-phenyl-propanoate 0 -(0 -NO
CI \
AY
Intermediate AY was prepared in analogy to Intermediate AP by using isopropyl (25)-2-(methylamino)-3-phenyl-propanoate (190 mg) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (220 mg, Intermediate AY) was obtained as light brown oil and used for the next step without further purification.
Intermediate AZ
(S)-tert-butyl 2-((chlorocarbonyl)(methyl)amino)-3-phenylpropanoate
CI \
AX
Intermediate AX was prepared in analogy to Intermediate AP by using (S)-ethy1-2-(methylamino)-3-phenylpropanoate instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (200 mg, Intermediate AX) was obtained as a light yellow oil and used for the next step without further purification Intermediate AY
Isopropyl (2S)-2-Ichlorocarbonyl(methyl)amino]-3-phenyl-propanoate 0 -(0 -NO
CI \
AY
Intermediate AY was prepared in analogy to Intermediate AP by using isopropyl (25)-2-(methylamino)-3-phenyl-propanoate (190 mg) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (220 mg, Intermediate AY) was obtained as light brown oil and used for the next step without further purification.
Intermediate AZ
(S)-tert-butyl 2-((chlorocarbonyl)(methyl)amino)-3-phenylpropanoate
- 87 -oX0 )-N 0 CI \
AZ
Step 1: Preparation of tert-butyl (2S)-2-(methylamino)-3-phenyl-propanoate (Compound AZ-1) OX
HNO
\
2-Methylpropene (25 g, 446 mmol) was bubbled into DCM (50 mL) at -78 C. Then the 2-methylpropene solution was added to a solution of (5)-2-(methylamino)-3-phenylpropanoic acid (500 mg) and H2SO4 (3.68 g, 2 mL) in dioxane (20 mL) at 0 C. The reaction mixture was stirred at room temperature for 18 hrs in a sealed tube.
The reaction mixture was poured into an ice cold aqueous KOH solution (8.4 g in water (30 mL)) and the resulting mixture was extracted with DCM (50 mL) twice. The organic layer was washed with brine (30 mL) 2 times, dried over Na2SO4 and concentrated in vacuo to afford tert-butyl (2S)-2-(methylamino)-3-phenyl-propanoate (710 mg, Compound AZ-1) as a light yellow oil.
Step 2: Preparation of (S)-tert-butyl 2-((chlorocarbonyl)(methyl)amino)-3-phenylpropanoate (Intermediate AZ) ,-N 0 CI \
AZ
AZ
Step 1: Preparation of tert-butyl (2S)-2-(methylamino)-3-phenyl-propanoate (Compound AZ-1) OX
HNO
\
2-Methylpropene (25 g, 446 mmol) was bubbled into DCM (50 mL) at -78 C. Then the 2-methylpropene solution was added to a solution of (5)-2-(methylamino)-3-phenylpropanoic acid (500 mg) and H2SO4 (3.68 g, 2 mL) in dioxane (20 mL) at 0 C. The reaction mixture was stirred at room temperature for 18 hrs in a sealed tube.
The reaction mixture was poured into an ice cold aqueous KOH solution (8.4 g in water (30 mL)) and the resulting mixture was extracted with DCM (50 mL) twice. The organic layer was washed with brine (30 mL) 2 times, dried over Na2SO4 and concentrated in vacuo to afford tert-butyl (2S)-2-(methylamino)-3-phenyl-propanoate (710 mg, Compound AZ-1) as a light yellow oil.
Step 2: Preparation of (S)-tert-butyl 2-((chlorocarbonyl)(methyl)amino)-3-phenylpropanoate (Intermediate AZ) ,-N 0 CI \
AZ
- 88 -Intermediate AZ was prepared in analogy to intermediate AP by using tert-butyl (2S)-2-(methylamino)-3-phenyl-propanoate (Compound AZ-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl (25)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (360 mg, Intermediate AZ) was obtained as a light yellow oil and used for next step without further purification Intermediate BA
N-P-Iacetyhmethyl)aminoiethyli-N-methyl-carbamoyl chloride N¨
O /¨
N
CI \
BA
Step 1: Preparation of tert-butyl N-P-Iacetyhmethyl)aminoiethyli-N-methyl-carbamate (Compound BA-1) of N¨
O /¨
N
--)-0 \
To a solution of tert-butyl methyl(2-(methylamino)ethyl)carbamate (1.13 g, 6 mmol) in pyridine (10 mL) was added acetic anhydride (3.06 g, 30 mmol) dropwise at 0 C. Then the solution was stirred at room temperature for 0.5 hr. The solvent was removed in vacuo and the residue was partitioned between Et0Ac (50 mL) and saturated aqueous NaHCO3 (25 mL). The organic layer was separated, washed with brine (20 mL), dried over Na2SO4 and concentrated in vacuo to afford tert-butyl N42-[acetyl(methyl)amino]ethyl]-N-methyl-carbamate (1.28 g, Compound BA-1) as a yellow oil.
N-P-Iacetyhmethyl)aminoiethyli-N-methyl-carbamoyl chloride N¨
O /¨
N
CI \
BA
Step 1: Preparation of tert-butyl N-P-Iacetyhmethyl)aminoiethyli-N-methyl-carbamate (Compound BA-1) of N¨
O /¨
N
--)-0 \
To a solution of tert-butyl methyl(2-(methylamino)ethyl)carbamate (1.13 g, 6 mmol) in pyridine (10 mL) was added acetic anhydride (3.06 g, 30 mmol) dropwise at 0 C. Then the solution was stirred at room temperature for 0.5 hr. The solvent was removed in vacuo and the residue was partitioned between Et0Ac (50 mL) and saturated aqueous NaHCO3 (25 mL). The organic layer was separated, washed with brine (20 mL), dried over Na2SO4 and concentrated in vacuo to afford tert-butyl N42-[acetyl(methyl)amino]ethyl]-N-methyl-carbamate (1.28 g, Compound BA-1) as a yellow oil.
- 89 -Step 2: Preparation of N-methyl-N-(2-(methylamino)ethyl)acetamide hydrochloride (Compound BA-2) N-H N/--/ HCI
\
A mixture of tert-butyl N[2-[acetyl(methyl)amino]ethy1]-N-methyl-carbamate (1.1 g, Compound BA-I) in HCl/Et0Ac (10 mL, 1N HC1 in Et0Ac) was stirred at room temperature for 2 hrs, then the mixture was filtered. The collected solid was washed with Et0Ac (5 mL) three times and dried in vacuo to afford the crude N-methyl-N-(2-(methylamino)ethyl)acetamide hydrochloride (460 mg, Compound BA-2) as a white solid.
Step 3: Preparation of N-I2-Iacetyhmethyl)aminolethyli-N-methyl-carbamoyl chloride (Intermediate BA) C:31 N-0 /¨/
N
CI \
BA
Intermediate BA was prepared in analogy to Intermediate AP by using N-methyl-N-(2-(methylamino)ethyl)acetamide hydrochloride (200 mg, Compound BA-2) instead of ethyl 2-(methylamino)acetate hydrochloride The crude N- [2-[acetyl(methyl)amino]ethyTh N-methyl-carbamoyl chloride (300 mg, Intermediate BA) was obtained and used for next step without further purification.
Intermediate BB
Methyl N- 12-Ichlorocarbonyl(methyl)amino]ethyli-N-methyl-carbamate
\
A mixture of tert-butyl N[2-[acetyl(methyl)amino]ethy1]-N-methyl-carbamate (1.1 g, Compound BA-I) in HCl/Et0Ac (10 mL, 1N HC1 in Et0Ac) was stirred at room temperature for 2 hrs, then the mixture was filtered. The collected solid was washed with Et0Ac (5 mL) three times and dried in vacuo to afford the crude N-methyl-N-(2-(methylamino)ethyl)acetamide hydrochloride (460 mg, Compound BA-2) as a white solid.
Step 3: Preparation of N-I2-Iacetyhmethyl)aminolethyli-N-methyl-carbamoyl chloride (Intermediate BA) C:31 N-0 /¨/
N
CI \
BA
Intermediate BA was prepared in analogy to Intermediate AP by using N-methyl-N-(2-(methylamino)ethyl)acetamide hydrochloride (200 mg, Compound BA-2) instead of ethyl 2-(methylamino)acetate hydrochloride The crude N- [2-[acetyl(methyl)amino]ethyTh N-methyl-carbamoyl chloride (300 mg, Intermediate BA) was obtained and used for next step without further purification.
Intermediate BB
Methyl N- 12-Ichlorocarbonyl(methyl)amino]ethyli-N-methyl-carbamate
- 90 -CI)L
BB
Step 1: Preparation of methyl N-methyl-N-I2-(methylamino)ethylicarbamate (Compound BB-1) N
H Nr.*---../ -----f 1 , 0 To a solution of N,/V'-dimethylethane-1,2-diamine (10 g) in THF (40 mL) was added methyl chloroformate (1.92 g) dropwise at -70 C in 1 hr. The mixture was stirred at 25 C
for 15 hrs and then filtered and washed with water and brine. The organic layer was dried and concentrated to afford a yellow residue, which was purified by column chromatography to afford methyl N-methyl-N-[2-(methylamino)ethyl]carbamate (2 g, Compound BB-1) as a colorless oil.
Step 2: Preparation of methyl N-I2-Ichlorocarbonyhmethyllaminolethyli-N-methyl-carbamate (Intermediate BB) CI)NV/
BB
Intermediate BB was prepared in analogy to Intermediate AP by using methyl N-methyl-N-[2-(methylamino)ethyl]carbamate (2.0 g, Compound BB-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude methyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (2.2 g, Intermediate BB) was obtained and used for next step without further purification.
Intermediate BC
BB
Step 1: Preparation of methyl N-methyl-N-I2-(methylamino)ethylicarbamate (Compound BB-1) N
H Nr.*---../ -----f 1 , 0 To a solution of N,/V'-dimethylethane-1,2-diamine (10 g) in THF (40 mL) was added methyl chloroformate (1.92 g) dropwise at -70 C in 1 hr. The mixture was stirred at 25 C
for 15 hrs and then filtered and washed with water and brine. The organic layer was dried and concentrated to afford a yellow residue, which was purified by column chromatography to afford methyl N-methyl-N-[2-(methylamino)ethyl]carbamate (2 g, Compound BB-1) as a colorless oil.
Step 2: Preparation of methyl N-I2-Ichlorocarbonyhmethyllaminolethyli-N-methyl-carbamate (Intermediate BB) CI)NV/
BB
Intermediate BB was prepared in analogy to Intermediate AP by using methyl N-methyl-N-[2-(methylamino)ethyl]carbamate (2.0 g, Compound BB-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude methyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (2.2 g, Intermediate BB) was obtained and used for next step without further purification.
Intermediate BC
- 91 -tert-Butyl N- 12-Ichlorocarbonyhmethyl)amino]ethy1]-N-methyl-carbamate Y
c, N¨
O /¨
N
CI \
BC
Step 1: Preparation of tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (Compound BC-1) Y
c, N¨
H N
\
To a solution of N,/V'-dimethylethane-1,2-diamine (40.4 g) in DCM (300 mL) was added a solution of Boc20 (10 g, 10.6 mL, 45.8 mmol) in DCM (100 mL) dropwise at 0 C
over 1 hr. The reaction mixture was stirred at room temperature for 18 hrs.
The organic layer was washed with saturated aqueous NaHCO3 (50 mL), brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography to afford tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (6.8 g, Compound BC-1) as a yellow oil. 1H NMR (400MHz, CDC13) 6 ppm: 3.34 (br. s., 2H), 2.89 (s, 3H), 2.74 (t, J
= 6.7 Hz, 2H), 2.46 (s, 3H), 1.47 (s, 9H).
Step 2: Preparation of tert-butyl N-12-Ichlorocarbonyhmethyl)amino]ethy1]-N-methyl-carbamate (Intermediate BC)
c, N¨
O /¨
N
CI \
BC
Step 1: Preparation of tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (Compound BC-1) Y
c, N¨
H N
\
To a solution of N,/V'-dimethylethane-1,2-diamine (40.4 g) in DCM (300 mL) was added a solution of Boc20 (10 g, 10.6 mL, 45.8 mmol) in DCM (100 mL) dropwise at 0 C
over 1 hr. The reaction mixture was stirred at room temperature for 18 hrs.
The organic layer was washed with saturated aqueous NaHCO3 (50 mL), brine (50 mL), dried over Na2SO4 and concentrated in vacuo. The residue was purified by column chromatography to afford tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (6.8 g, Compound BC-1) as a yellow oil. 1H NMR (400MHz, CDC13) 6 ppm: 3.34 (br. s., 2H), 2.89 (s, 3H), 2.74 (t, J
= 6.7 Hz, 2H), 2.46 (s, 3H), 1.47 (s, 9H).
Step 2: Preparation of tert-butyl N-12-Ichlorocarbonyhmethyl)amino]ethy1]-N-methyl-carbamate (Intermediate BC)
- 92 _ Y
C) Or /¨
N
CI \
BC
Intermediate BC was prepared in analogy to Intermediate AP by using tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (1.15 g, Compound BC-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (1.3 g, Intermediate BC) was obtained and used for the next step without further purification.
Intermediate BD
Ethyl N- 12-Ichlorocarbonyl(methyl)amino]ethyli-N-methyl-carbamate )L N 0 CI NI V/
I
BD
Step 1: Preparation of ethyl N-methyl-N-I2-(methylamino)ethylicarbamate (Compound BD-1) I
N
H N= rC) I
To a solution of N,/V'-dimethylethane-1,2-diamine (10 g) in DCM (40 mL) was added ethyl chloroformate (2.58 g) dropwise at -70 C in 1 hr. The reaction mixture was stirred at 25 C for 15 hrs and then filtered and washed with water and brine.
The organic layer was dried and concentrated in vacuo. The yellow residue was purified by column
C) Or /¨
N
CI \
BC
Intermediate BC was prepared in analogy to Intermediate AP by using tert-butyl N-methyl-N-[2-(methylamino)ethyl]carbamate (1.15 g, Compound BC-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude tert-butyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (1.3 g, Intermediate BC) was obtained and used for the next step without further purification.
Intermediate BD
Ethyl N- 12-Ichlorocarbonyl(methyl)amino]ethyli-N-methyl-carbamate )L N 0 CI NI V/
I
BD
Step 1: Preparation of ethyl N-methyl-N-I2-(methylamino)ethylicarbamate (Compound BD-1) I
N
H N= rC) I
To a solution of N,/V'-dimethylethane-1,2-diamine (10 g) in DCM (40 mL) was added ethyl chloroformate (2.58 g) dropwise at -70 C in 1 hr. The reaction mixture was stirred at 25 C for 15 hrs and then filtered and washed with water and brine.
The organic layer was dried and concentrated in vacuo. The yellow residue was purified by column
- 93 -chromatography to afford ethyl N-methyl-N-[2-(methylamino)ethyl]carbamate (2 g, Compound BD-1) as a colorless oil.
Step 2: Preparation of ethyl N-I2-Ichlorocarbonyhmethyllaminolethyli-N-methyl-carbamate (Intermediate BD) )L N 0 CINIV
I
BD
Intermediate BD was prepared in analogy to Intermediate AA by using ethyl N-methyl-N-[2-(methylamino)ethyl]carbamate (Compound BD-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (2.2 g, Intermediate BD) was obtained and used for the next step without further purification.
Intermediate BE
2-IChlorocarbonyhmethyllaminolethyl N-butyl-N-methyl-carbamate cio 0 r V
I
BE
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BE-1) OH
r c)2N
I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was
Step 2: Preparation of ethyl N-I2-Ichlorocarbonyhmethyllaminolethyli-N-methyl-carbamate (Intermediate BD) )L N 0 CINIV
I
BD
Intermediate BD was prepared in analogy to Intermediate AA by using ethyl N-methyl-N-[2-(methylamino)ethyl]carbamate (Compound BD-1) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude ethyl N-[2-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (2.2 g, Intermediate BD) was obtained and used for the next step without further purification.
Intermediate BE
2-IChlorocarbonyhmethyllaminolethyl N-butyl-N-methyl-carbamate cio 0 r V
I
BE
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BE-1) OH
r c)2N
I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was
- 94 -added Boc20 (34.87 g, 159.77 mmol) at 25 C. The mixture was stirred at 25 C
for 16 hrs and then concentrated. The residue was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BE-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyl(methyl)amino] ethyl N-butyl-N-methyl-carbamate (Compound BE-2) I
-- ,N........7"--0)LNV......--7.......' To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (880 mg, Compound BE-1) and Et3N (1 g, 10.08 mmol) in DCM (10 mL) was added N-butyl-N-methyl-carbamoyl chloride (903 mg, 7.04 mmol) dropwise at -10 C in 1 hr. The reaction mixture was stirred at 25 C for 15 hrs and then filtered and washed with water and brine.
The organic layer was dried and concentrated to afford 2-[tert-butoxycarbonyl(methyl)amino] ethyl N-butyl-N-methyl-carbamate (2 g, Compound BE-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl N-butyl-N-methyl-carbamate hydrochloride (Compound BE-3) H
, HCI
To a solution of 2-[tert-butoxycarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (1 g, Compound BE-2) was added HC1/EA (40 mL, 1M). The reaction mixture was stirred at 0 C for 0.5 hr and warmed to 25 C and stirred for another 15.5 hrs. The reaction mixture was concentrated to afford 2-(methylamino)ethyl-N-butyl-N-methyl-carbamate hydrochloride (400 mg, Compound BE-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyl(methyl)amino] ethyl N-butyl-N-methyl-
for 16 hrs and then concentrated. The residue was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BE-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyl(methyl)amino] ethyl N-butyl-N-methyl-carbamate (Compound BE-2) I
-- ,N........7"--0)LNV......--7.......' To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (880 mg, Compound BE-1) and Et3N (1 g, 10.08 mmol) in DCM (10 mL) was added N-butyl-N-methyl-carbamoyl chloride (903 mg, 7.04 mmol) dropwise at -10 C in 1 hr. The reaction mixture was stirred at 25 C for 15 hrs and then filtered and washed with water and brine.
The organic layer was dried and concentrated to afford 2-[tert-butoxycarbonyl(methyl)amino] ethyl N-butyl-N-methyl-carbamate (2 g, Compound BE-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl N-butyl-N-methyl-carbamate hydrochloride (Compound BE-3) H
, HCI
To a solution of 2-[tert-butoxycarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (1 g, Compound BE-2) was added HC1/EA (40 mL, 1M). The reaction mixture was stirred at 0 C for 0.5 hr and warmed to 25 C and stirred for another 15.5 hrs. The reaction mixture was concentrated to afford 2-(methylamino)ethyl-N-butyl-N-methyl-carbamate hydrochloride (400 mg, Compound BE-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyl(methyl)amino] ethyl N-butyl-N-methyl-
- 95 -carbamate (Intermediate BE) r V
I
BE
Intermediate BE was prepared in analogy to Intermediate AP by using 2-(methylamino)ethyl N-butyl-N-methyl-carbamate hydrochloride (374 mg, Compound BE-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (330 mg, Intermediate BE) was obtained and used for next step without further purification.
Intermediate BF
2-1Chlorocarbonyl(methyl)aminoiethyl pyrrolidine-l-carboxylate a 0 0 r V
BF
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BF-1) OH
(11 r c>2-N
I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The mixture was stirred at 25 C
for 16 hrs.
The reaction mixture was concentrated to give the residue, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BF-1) as a colorless oil.
I
BE
Intermediate BE was prepared in analogy to Intermediate AP by using 2-(methylamino)ethyl N-butyl-N-methyl-carbamate hydrochloride (374 mg, Compound BE-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (330 mg, Intermediate BE) was obtained and used for next step without further purification.
Intermediate BF
2-1Chlorocarbonyl(methyl)aminoiethyl pyrrolidine-l-carboxylate a 0 0 r V
BF
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BF-1) OH
(11 r c>2-N
I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The mixture was stirred at 25 C
for 16 hrs.
The reaction mixture was concentrated to give the residue, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BF-1) as a colorless oil.
- 96 -Step 2: Preparation of 2-Itert-butoxycarbonyhmethyllamino] ethyl pyrrolidine-l-carboxylate (Compound BF-2) OC) 0 11\1 0)L NO
To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (300 mg, 1.71 mmol, Compound BF-1) and Et3N (578 mg, 5.71 mmol) in DCM (5 mL) was added pyrrolidine- 1 -carbonyl chloride (458 mg, 3.4 mmol) dropwise at 0 C for 0.5 hr and then stirred at 25 C for 15.5 hrs. After filtration, the filtrate was washed with water and brine.
The organic layer was dried and concentrated to afford the 2-[tert-butoxycarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (335 mg, Compound BF-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl pyrrolidine-l-carboxylate hydrochloride (Compound BF-3) H
V
HCI
2-[tert-butoxycarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (335 mg, Compound BF-2) was added to HC1 in EA (12.3 mL, 1M) and the mixture was stirred at 0 C for 0.5 hr and then at 25 C for another 15.5 hrs. The reaction mixture was concentrated to afford 2-(methylamino)ethyl pyrrolidine- 1 -carboxylate hydrochloride (300 mg, Compound BF-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyllamino] ethyl pyrrolidine-1-carboxylate (Intermediate BF)
To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (300 mg, 1.71 mmol, Compound BF-1) and Et3N (578 mg, 5.71 mmol) in DCM (5 mL) was added pyrrolidine- 1 -carbonyl chloride (458 mg, 3.4 mmol) dropwise at 0 C for 0.5 hr and then stirred at 25 C for 15.5 hrs. After filtration, the filtrate was washed with water and brine.
The organic layer was dried and concentrated to afford the 2-[tert-butoxycarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (335 mg, Compound BF-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl pyrrolidine-l-carboxylate hydrochloride (Compound BF-3) H
V
HCI
2-[tert-butoxycarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (335 mg, Compound BF-2) was added to HC1 in EA (12.3 mL, 1M) and the mixture was stirred at 0 C for 0.5 hr and then at 25 C for another 15.5 hrs. The reaction mixture was concentrated to afford 2-(methylamino)ethyl pyrrolidine- 1 -carboxylate hydrochloride (300 mg, Compound BF-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyllamino] ethyl pyrrolidine-1-carboxylate (Intermediate BF)
- 97 -BF
Intermediate BF was prepared in analogy to Intermediate AP by using the 2-(methylamino)ethyl pyrrolidine- 1 -carboxylate hydrochloride (299 mg, Compound BF-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (230 mg, Intermediate BF) was obtained and used for next step without further purification.
Intermediate BG
2-1Chlorocarbonyl(methyl)aminolethyl N-methyl-N-propyl-carbamate Ci0 0 r BG
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BG-1) OH
02Nr I
BC-I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The reaction mixture was stirred at 25 C for 16 hrs, then concentrated to give the residue, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BG-1) as a colorless oil.
Intermediate BF was prepared in analogy to Intermediate AP by using the 2-(methylamino)ethyl pyrrolidine- 1 -carboxylate hydrochloride (299 mg, Compound BF-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (230 mg, Intermediate BF) was obtained and used for next step without further purification.
Intermediate BG
2-1Chlorocarbonyl(methyl)aminolethyl N-methyl-N-propyl-carbamate Ci0 0 r BG
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BG-1) OH
02Nr I
BC-I
To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The reaction mixture was stirred at 25 C for 16 hrs, then concentrated to give the residue, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BG-1) as a colorless oil.
- 98 -Step 2: Preparation of tert-butyl-N-methyl-N-12-Imethyl(propyl)carbamoyl]oxyethyl]
carbamate (Compound BG-2) To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (265 mg, Compound BG-1) and Et3N (1 mL, 5.71 mmol) in DCM (5 mL) was added N-methyl-N-propyl-carbamoyl chloride (410 mg, 1.83 mmol) dropwise at 0 C for 0.5 hr. The reaction mixture was stirred at 25 C for 15.5 hrs and then filtered and the filtrate was washed with water and brine. The organic layer was dried and concentrated to afford tert-butyl N-methyl-N-[2-[methyl(propyl)carbamoyl]oxyethyl]carbamate (380 mg, Compound BG-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (Compound BG-3) )LNV\V
HCI
tert-butyl N-methyl-N-[2-[methyl(propyl)carbamoyl]oxyethyl]carbamate (380 mg, Compound BG-2) was added to HC1 in EA (13.7 mL, 1M). The mixture was stirred at 0 C
for 0.5 hr. Then the mixture was stirred at 25 C for another 15.5 hrs and concentrated to afford 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (300 mg, Compound BG-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyl)amino] ethyl N-methyl-N-propyl-carbamate (Intermediate BG)
carbamate (Compound BG-2) To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (265 mg, Compound BG-1) and Et3N (1 mL, 5.71 mmol) in DCM (5 mL) was added N-methyl-N-propyl-carbamoyl chloride (410 mg, 1.83 mmol) dropwise at 0 C for 0.5 hr. The reaction mixture was stirred at 25 C for 15.5 hrs and then filtered and the filtrate was washed with water and brine. The organic layer was dried and concentrated to afford tert-butyl N-methyl-N-[2-[methyl(propyl)carbamoyl]oxyethyl]carbamate (380 mg, Compound BG-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (Compound BG-3) )LNV\V
HCI
tert-butyl N-methyl-N-[2-[methyl(propyl)carbamoyl]oxyethyl]carbamate (380 mg, Compound BG-2) was added to HC1 in EA (13.7 mL, 1M). The mixture was stirred at 0 C
for 0.5 hr. Then the mixture was stirred at 25 C for another 15.5 hrs and concentrated to afford 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (300 mg, Compound BG-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyl)amino] ethyl N-methyl-N-propyl-carbamate (Intermediate BG)
- 99 -cio 0 r )L yz V
BG
Intermediate BG was prepared in analogy to Intermediate AP by using 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (330 mg, Compound BG-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The 2-[chlorocarbonyl(methyl)amino]ethyl-N-methyl-N-propyl-carbamate (300 mg, Intermediate BG) was obtained and used for next step without further purification.
Intermediate BH
2-1Chlorocarbonyl(methyl)aminolethyl N,N-diethylcarbamate r V
L-, BH
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BH-1) OH
i), r To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The mixture was stirred at 25 C
for 16 hrs and then concentrated, the residue was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BH-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyl(methyl)aminolethyl-/V,N-diethylcarbamate (Compound BH-2)
BG
Intermediate BG was prepared in analogy to Intermediate AP by using 2-(methylamino)ethyl N-methyl-N-propyl-carbamate hydrochloride (330 mg, Compound BG-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The 2-[chlorocarbonyl(methyl)amino]ethyl-N-methyl-N-propyl-carbamate (300 mg, Intermediate BG) was obtained and used for next step without further purification.
Intermediate BH
2-1Chlorocarbonyl(methyl)aminolethyl N,N-diethylcarbamate r V
L-, BH
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BH-1) OH
i), r To a solution of 2-(methylamino)ethanol (10 g, 133.14 mmol) in DCM (10 mL) was added Boc20 (34.87 g, 159.77 mmol) at 25 C. The mixture was stirred at 25 C
for 16 hrs and then concentrated, the residue was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (20 g, Compound BH-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyl(methyl)aminolethyl-/V,N-diethylcarbamate (Compound BH-2)
- 100 -0.......0 0 i --1 0 )1' L--..N Vs.'s' To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (200 mg, 1.14 mmol, Compound BH-1) and Et3N (578 mg, 5.71 mmol) in DCM (5 mL) was added 1V,N-diethylcarbamoyl chloride (248 mg, 1.83 mmol) dropwise at 0 C for 0.5 hr and stirred at 25 C for 15.5 hrs. After filtration, the filtrate was washed with water and brine. The organic layer was dried and concentrated to afford the 2-[tert-butoxycarbonyl(methyl)amino]ethyl N,N-diethylcarbamate (313 mg, Compound BH-2) as a colorless oil.
Step 3: Preparation of 2-(methylamino)ethyl /V,N-diethylcarbamate hydrochloride (Compound BH-3) H
N.,...7"--.0)*LN7-......"
H C I
2-[tert-butoxycarbonyl(methyl)amino]ethyl /V,N-diethylcarbamate (436 mg, 1.77 mmol, Compound BH-2) was added to HC1 in EA (17 mL, 1M). The mixture was stirred at 0 C for 0.5 hr. Then the mixture was stirred at 25 C for another 15.5 hrs and concentrated to afford 2-(methylamino)ethyl /V,N-diethylcarbamate hydrochloride (230 mg, Compound BH-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyllaminolethyl /V,N-diethylcarbamate (Intermediate BH) Cio o I
BH
Step 3: Preparation of 2-(methylamino)ethyl /V,N-diethylcarbamate hydrochloride (Compound BH-3) H
N.,...7"--.0)*LN7-......"
H C I
2-[tert-butoxycarbonyl(methyl)amino]ethyl /V,N-diethylcarbamate (436 mg, 1.77 mmol, Compound BH-2) was added to HC1 in EA (17 mL, 1M). The mixture was stirred at 0 C for 0.5 hr. Then the mixture was stirred at 25 C for another 15.5 hrs and concentrated to afford 2-(methylamino)ethyl /V,N-diethylcarbamate hydrochloride (230 mg, Compound BH-3) as a colorless oil.
Step 4: Preparation of 2-Ichlorocarbonyhmethyllaminolethyl /V,N-diethylcarbamate (Intermediate BH) Cio o I
BH
- 101 -Intermediate BH was prepared in analogy to Intermediate AP by using 2-(methylamino)ethyl /V,N-diethylcarbamate hydrochloride (274 mg, Compound BH-3) instead of ethyl 2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl /V,N-diethylcarbamate (250 mg, Intermediate BH) was obtained and used for next step without further purification.
Intermediate BI
2-IChlorocarbonyhmethyl)aminolethyl ethyl carbonate cio r 0 No)-L0/\
BI
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BI-1) .0y0 N.V. OH
To a solution of 2-(methylamino)ethanol (1 g, 13.31 mmol) in DCM (10 mL) was added Boc20 (3.49 g, 15.98 mmol) at 25 C. The reaction mixture was stirred at 25 C for 16 hrs, then concentrated to give the crude product, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (1.6 g, Compound BI-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyhmethyl)aminolethyl methyl carbonate (Compound BI-2)
Intermediate BI
2-IChlorocarbonyhmethyl)aminolethyl ethyl carbonate cio r 0 No)-L0/\
BI
Step 1: Preparation of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (Compound BI-1) .0y0 N.V. OH
To a solution of 2-(methylamino)ethanol (1 g, 13.31 mmol) in DCM (10 mL) was added Boc20 (3.49 g, 15.98 mmol) at 25 C. The reaction mixture was stirred at 25 C for 16 hrs, then concentrated to give the crude product, which was purified by column chromatography to afford tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (1.6 g, Compound BI-1) as a colorless oil.
Step 2: Preparation of 2-Itert-butoxycarbonyhmethyl)aminolethyl methyl carbonate (Compound BI-2)
- 102 -,0 0 ...---- y 0 N.........õ,-......0)...0 To a solution of tert-butyl N-(2-hydroxyethyl)-N-methyl-carbamate (1 g, Compound BI-1), DMAP (0.1 g) and pyridine (1.15 g, 11.41 mmol) in EA (20 mL) was added methyl chloroformate (1.21 g, 11.15 mmol) dropwise at -10 C. The mixture was stirred at -10 C for 1 hr. The reaction mixture was filtered and the filtrate was washed with 5% citric acid and brine. The organic layer was dried and concentrated to afford 2-[tert-butoxycarbonyl(methyl)amino]ethyl methyl carbonate (1.22 g, Compound BI-2) as a colorless oil.
Step 3: Preparation of ethyl 2-(methylamino)ethyl carbonate hydrochloride (Compound BI-3) H
(:)).L N o/
HCI
2-[tert-butoxycarbonyl(methyl)amino]ethyl methyl carbonate (1.22 g, 4.94 mmol, Compound BI-2) was added to HC1 in EA (10 mL, 40 mmol) and the mixture was stirred at 0 C for 0.5 hr and at 25 C for another 15.5 hrs. The reaction mixture was concentrated to afford ethyl 2-(methylamino)ethyl carbonate hydrochloride (1.06 g, Compound BI-3).
Step 4: Preparation of 2-Ichlorocarbonyhmethyllaminolethyl ethyl carbonate (Intermediate BI) CIO
r 0 NoA0/\
BI
Intermediate BI was prepared in analogy to Intermediate AP by using ethyl 2-(methylamino)ethyl carbonate hydrochloride (150 mg, Intermediate BI-3) instead of ethyl
Step 3: Preparation of ethyl 2-(methylamino)ethyl carbonate hydrochloride (Compound BI-3) H
(:)).L N o/
HCI
2-[tert-butoxycarbonyl(methyl)amino]ethyl methyl carbonate (1.22 g, 4.94 mmol, Compound BI-2) was added to HC1 in EA (10 mL, 40 mmol) and the mixture was stirred at 0 C for 0.5 hr and at 25 C for another 15.5 hrs. The reaction mixture was concentrated to afford ethyl 2-(methylamino)ethyl carbonate hydrochloride (1.06 g, Compound BI-3).
Step 4: Preparation of 2-Ichlorocarbonyhmethyllaminolethyl ethyl carbonate (Intermediate BI) CIO
r 0 NoA0/\
BI
Intermediate BI was prepared in analogy to Intermediate AP by using ethyl 2-(methylamino)ethyl carbonate hydrochloride (150 mg, Intermediate BI-3) instead of ethyl
- 103 -2-(methylamino)acetate hydrochloride. The crude 2-[chlorocarbonyl(methyl)amino]ethyl ethyl carbonate (145 mg, Intermediate BI) was obtained and used for next step without further purification.
PREPARATIVE EXAMPLES
Example 1 6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj \
NN
µ, ......
S N N
NNNH
.
Method A:
Step 1: Preparation of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la) NH
I )=() la To a solution of aminomalononitrile p-toluenesulfonate (25 g, 98.5 mmol, TCI, Catalog number: A1119-25G) in dry THF (100 mL) was added benzyl isocyanate (13.2 g, 98.5 mmol) and TEA (10.2 g, 79.0 mmol) at RT. After stirred at RT for 24 hrs, the reaction was concentrated in vacuo and the residue was partitioned between Et0Ac (500 mL) and water (250 mL). The separated organic layer was washed with brine (50 mL) twice, and
PREPARATIVE EXAMPLES
Example 1 6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj \
NN
µ, ......
S N N
NNNH
.
Method A:
Step 1: Preparation of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la) NH
I )=() la To a solution of aminomalononitrile p-toluenesulfonate (25 g, 98.5 mmol, TCI, Catalog number: A1119-25G) in dry THF (100 mL) was added benzyl isocyanate (13.2 g, 98.5 mmol) and TEA (10.2 g, 79.0 mmol) at RT. After stirred at RT for 24 hrs, the reaction was concentrated in vacuo and the residue was partitioned between Et0Ac (500 mL) and water (250 mL). The separated organic layer was washed with brine (50 mL) twice, and
- 104 -extracted with sodium hydroxide solution (50 mL, 1/V) twice. The combined sodium hydroxide solution layer was neutralized with 10 wt.% sodium hydrogen sulfate solution and extracted with Et0Ac. The separated organic layer was washed with brine, dried over Na2SO4, filtered and concentrated in vacuo. The residue was triturated in 2-isopropoxypropane and then the suspension was filtered to give 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (15 g, Compound la) as a yellow solid. The product was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]:
215.
Step 2: Preparation of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) H
N----", N\
, 1 /=
HSN.----N
=
lb To a solution of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (15.0 g, 70.0 mmol, Compound la) in THF (700 mL) was added benzoylisothiocyanate (28.6 g, 175.1 mmol, TCI, Catalog number: A11596-100G) dropwise. After stirred at RT for 12 hrs, the reaction mixture was concentrated in vacuo. The residue was triturated in diethyl ether (100 mL) and the resulting precipitate was collected by filtration.
To a solution of the obtained precipitate in THF (700 mL) was added sodium hydroxide (70 mL, 2 N) . The mixture was refluxed for 50 hrs, and then acidified to pH=3 with 10 wt.% aqueous sodium hydrogen sulfate solution. The resulting precipitate was collected by filtration to give a crude 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (8.1 g, Compound lb) as a yellow solid. The product was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]: 274.
Step 3: Preparation of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound lc)
215.
Step 2: Preparation of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) H
N----", N\
, 1 /=
HSN.----N
=
lb To a solution of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (15.0 g, 70.0 mmol, Compound la) in THF (700 mL) was added benzoylisothiocyanate (28.6 g, 175.1 mmol, TCI, Catalog number: A11596-100G) dropwise. After stirred at RT for 12 hrs, the reaction mixture was concentrated in vacuo. The residue was triturated in diethyl ether (100 mL) and the resulting precipitate was collected by filtration.
To a solution of the obtained precipitate in THF (700 mL) was added sodium hydroxide (70 mL, 2 N) . The mixture was refluxed for 50 hrs, and then acidified to pH=3 with 10 wt.% aqueous sodium hydrogen sulfate solution. The resulting precipitate was collected by filtration to give a crude 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (8.1 g, Compound lb) as a yellow solid. The product was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]: 274.
Step 3: Preparation of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound lc)
- 105 -H
1 >0 SN..........N
lc To a solution of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (5.46 g, 20.0 mmol, Compound lb) in DMF was added potassium carbonate (2.76 g, 20.0 mmol). And then 1-bromopropane (2.44 g, 20.0 mmol, TCI, Catalog number: B0638-500G) in DMF (5.0 mL) was slowly added to previous solution. After stirred at RT for 12 hrs, the reaction mixture was poured into water (200 mL), then acidified with 10 wt.% aqueous sodium hydrogen sulfate solution and extracted with Et0Ac (100 mL) twice. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo to give the crude product, which was purified by flash chromatography on silica gel to give 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (4.8 g, Compound 1c) as a white solid. MS obsd.
(ESL') [(M+H)+] : 316.
Step 4: Preparation of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d) H
N.---N
0 >0 \\ ..........N
N
ld To a suspension of compound 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (2.7 g, 8.7 mmol, Compound 1c) in DCM/Me0H (500 mL, V/V = 1:1) was added 3-chloroperbenzoic acid (2.15 g, 8.7 mmol, 70% purity, Aldrich, Catalog number:
100G). After reaction mixture was stirred for 2 hrs, the volume of reaction mixture was reduced in vacuo to about 50 mL. The resulting precipitate was collected by filtration, washed with methanol and dried to give 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-
1 >0 SN..........N
lc To a solution of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (5.46 g, 20.0 mmol, Compound lb) in DMF was added potassium carbonate (2.76 g, 20.0 mmol). And then 1-bromopropane (2.44 g, 20.0 mmol, TCI, Catalog number: B0638-500G) in DMF (5.0 mL) was slowly added to previous solution. After stirred at RT for 12 hrs, the reaction mixture was poured into water (200 mL), then acidified with 10 wt.% aqueous sodium hydrogen sulfate solution and extracted with Et0Ac (100 mL) twice. The organic layer was washed with brine, dried over Na2SO4 and concentrated in vacuo to give the crude product, which was purified by flash chromatography on silica gel to give 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (4.8 g, Compound 1c) as a white solid. MS obsd.
(ESL') [(M+H)+] : 316.
Step 4: Preparation of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d) H
N.---N
0 >0 \\ ..........N
N
ld To a suspension of compound 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (2.7 g, 8.7 mmol, Compound 1c) in DCM/Me0H (500 mL, V/V = 1:1) was added 3-chloroperbenzoic acid (2.15 g, 8.7 mmol, 70% purity, Aldrich, Catalog number:
100G). After reaction mixture was stirred for 2 hrs, the volume of reaction mixture was reduced in vacuo to about 50 mL. The resulting precipitate was collected by filtration, washed with methanol and dried to give 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-
- 106 -one (1.0 g, Compound 1d) as a white solid. The product was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]: 332.
Step 5: Preparation of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) H
N----"N
0 1 >0 \\ .........N
H N=S N
le To a solution of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (1.52 g, 4.6 mmol, Compound 1d) in Eaton's reagent (40 mL, phosphorus pentoxide, 7.5 wt. %
in methanesulphonic acid, Aldrich, Catalog number: 380814-100ML) was added sodium azide (360 mg, 5.5 mmol) at 50 C. After being stirred at this temperature for 30 minutes, the reaction mixture was cooled to RT and poured into sat. aqueous sodium bicarbonate solution. The reaction mixture was extracted with n-BuOH (100 mL) twice, and the organic phase was concentrated in vacuo. The residue was submitted for purification by prep-HPLC to give 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (1.2 g, Compound le) as a white solid. 'H NMR (400 MHz, DMSO-d6) 6 ppm: 10.65 (br. s., 1H), 7.26-7.37 (m, 5H), 6.98 (br. s., 2H), 4.97 (s, 2H), 4.02 (s, 1H), 3.33 (t, J=
7.53 Hz, 2H), 1.55-1.74 (m, 2H), 0.92 (t, J=7.53 Hz, 3H). MS obsd. (ESE) [(M+1-1) ]: 347.
Separation of compound le by chiral HPLC afforded Compound le-A (slower eluting, 500 mg) and Compound le-B (faster eluting, 490 mg) as white solid.
(Separation condition: methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AS-3 column.) Compound le-A: 1H NMR (DMSO-d6, 400 MHz) gppm: 10.56 (s, 1H), 7.21 - 7.46 (m, 5H), 7.03 (s, 2H), 4.96 (s, 2H), 4.04 (s, 1H), 3.25 - 3.33 (m, 2H), 1.59 -1.67 (m, 2H), 0.92 (t, J= 7.4 Hz, 3H).
Compound le-B: 1H NMR (DMSO-d6, 400 MHz) gppm: 10.57 (s, 1H), 7.23 - 7.39 (m, 5H), 6.97 (s, 2H), 4.96 (s, 2H), 4.05 (s, 1H), 3.31 - 3.30 (m, 2H), 1.49 -1.74 (m, 2H), 0.91 (t, J= 7.4 Hz, 3H).
Step 5: Preparation of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) H
N----"N
0 1 >0 \\ .........N
H N=S N
le To a solution of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (1.52 g, 4.6 mmol, Compound 1d) in Eaton's reagent (40 mL, phosphorus pentoxide, 7.5 wt. %
in methanesulphonic acid, Aldrich, Catalog number: 380814-100ML) was added sodium azide (360 mg, 5.5 mmol) at 50 C. After being stirred at this temperature for 30 minutes, the reaction mixture was cooled to RT and poured into sat. aqueous sodium bicarbonate solution. The reaction mixture was extracted with n-BuOH (100 mL) twice, and the organic phase was concentrated in vacuo. The residue was submitted for purification by prep-HPLC to give 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (1.2 g, Compound le) as a white solid. 'H NMR (400 MHz, DMSO-d6) 6 ppm: 10.65 (br. s., 1H), 7.26-7.37 (m, 5H), 6.98 (br. s., 2H), 4.97 (s, 2H), 4.02 (s, 1H), 3.33 (t, J=
7.53 Hz, 2H), 1.55-1.74 (m, 2H), 0.92 (t, J=7.53 Hz, 3H). MS obsd. (ESE) [(M+1-1) ]: 347.
Separation of compound le by chiral HPLC afforded Compound le-A (slower eluting, 500 mg) and Compound le-B (faster eluting, 490 mg) as white solid.
(Separation condition: methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AS-3 column.) Compound le-A: 1H NMR (DMSO-d6, 400 MHz) gppm: 10.56 (s, 1H), 7.21 - 7.46 (m, 5H), 7.03 (s, 2H), 4.96 (s, 2H), 4.04 (s, 1H), 3.25 - 3.33 (m, 2H), 1.59 -1.67 (m, 2H), 0.92 (t, J= 7.4 Hz, 3H).
Compound le-B: 1H NMR (DMSO-d6, 400 MHz) gppm: 10.57 (s, 1H), 7.23 - 7.39 (m, 5H), 6.97 (s, 2H), 4.96 (s, 2H), 4.05 (s, 1H), 3.31 - 3.30 (m, 2H), 1.49 -1.74 (m, 2H), 0.91 (t, J= 7.4 Hz, 3H).
- 107 -Step 6: Preparation of 6-amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (Example 1) 0 rj NH2 ...-N
1 \
N"N
NµSµ( N
N ----NH
To a solution of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (300 mg, Compound le), pyridine (329 mg, 4.2 mmol) and DIPEA (538 mg, 4.2 mmol) in NMP (5 mL) was added N-methyl-N-propyl-carbamoyl chloride (564 mg, 4.2 mmol, Intermediate AA) at RT. The mixture was stirred at RT for 10 hrs. The reaction mixture was concentrated and the residue was purified by prep-HPLC to give 6-amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (108 mg, Example 1) as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.45 - 7.24 (m, 5H), 6.89 (s, 2H), 5.01 (s, 2H), 4.17 (s, 1H), 3.44 - 3.34 (m, 2H), 3.36 - 3.34 (m, 2H), 3.10 - 3.00 (m, 3H), 1.74 - 1.52 (m, 4H), 1.01 - 0.72 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Separation of compound of Example 1 by chiral HPLC afforded Example 1-A
(slower eluting, 50 mg) and Example 1-B (faster eluting, 40 mg) as white solid with isopropanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Example 1-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.44-7.24 (m, 5H), 6.89 (s, 2H), 5.01 (s, 2H), 4.17 (s, 1H), 3.44-3.37 (m, 2H), 3.37-3.35 (m, 2H), 3.10-3.00 (m, 3H), 1.74-1.52 (m, 4H), 1.00-0.72 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Example 1-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.45-7.26 (m, 5H), 6.88 (s, 2H), 5.01 (s, 2H), 4.15 (s, 1H), 3.44-3.36 (m, 2H), 3.34 (s, 2H), 3.10-3.01 (m, 3H), 1.77-1.52 (m, 4H), 1.02-0.67 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Method B: Alternative method to prepare 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le)
1 \
N"N
NµSµ( N
N ----NH
To a solution of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (300 mg, Compound le), pyridine (329 mg, 4.2 mmol) and DIPEA (538 mg, 4.2 mmol) in NMP (5 mL) was added N-methyl-N-propyl-carbamoyl chloride (564 mg, 4.2 mmol, Intermediate AA) at RT. The mixture was stirred at RT for 10 hrs. The reaction mixture was concentrated and the residue was purified by prep-HPLC to give 6-amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (108 mg, Example 1) as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.45 - 7.24 (m, 5H), 6.89 (s, 2H), 5.01 (s, 2H), 4.17 (s, 1H), 3.44 - 3.34 (m, 2H), 3.36 - 3.34 (m, 2H), 3.10 - 3.00 (m, 3H), 1.74 - 1.52 (m, 4H), 1.01 - 0.72 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Separation of compound of Example 1 by chiral HPLC afforded Example 1-A
(slower eluting, 50 mg) and Example 1-B (faster eluting, 40 mg) as white solid with isopropanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Example 1-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.44-7.24 (m, 5H), 6.89 (s, 2H), 5.01 (s, 2H), 4.17 (s, 1H), 3.44-3.37 (m, 2H), 3.37-3.35 (m, 2H), 3.10-3.00 (m, 3H), 1.74-1.52 (m, 4H), 1.00-0.72 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Example 1-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.45-7.26 (m, 5H), 6.88 (s, 2H), 5.01 (s, 2H), 4.15 (s, 1H), 3.44-3.36 (m, 2H), 3.34 (s, 2H), 3.10-3.01 (m, 3H), 1.77-1.52 (m, 4H), 1.02-0.67 (m, 6H). MS obsd. (ESI+) [(M+H)+]: 446.
Method B: Alternative method to prepare 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le)
- 108 -H
NN
"
H N=S N-----N
le Step 1: Preparation of N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (Compound if) CI
N
I
H
if To a solution of 4,6-dichloro-5-nitro-2-propylsulfanylpyrimidine (150.0 g, 559.5 mmol) and DIPEA (108.5 g, 839.2 mmol) in THF(1.5 L) was added phenylmethanamine (60.0 g, 559.5 mmol) in THF(200 mL) slowly at -78 C. After addition, the mixture was warmed to 25 C, and stirred at this temperature for 16 hrs. The resulting mixture was diluted with EA (1 L), washed with water (400 mL) three times and brine (500 mL). The separated organic phase was dried over Na2SO4, filtered and concentrated in vacuo to give N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (180.0 g, Compound lf) as a yellow solid and used for next step without further purification. MS
obsd. (ESI ) [(M+H)+]: 339.1.
Step 2: Preparation of N4-benzy1-6-chloro-2-propylsulfanyl-pyrimidine-4,5-diamine (Compound 1g) CI
, I
H
lg
NN
"
H N=S N-----N
le Step 1: Preparation of N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (Compound if) CI
N
I
H
if To a solution of 4,6-dichloro-5-nitro-2-propylsulfanylpyrimidine (150.0 g, 559.5 mmol) and DIPEA (108.5 g, 839.2 mmol) in THF(1.5 L) was added phenylmethanamine (60.0 g, 559.5 mmol) in THF(200 mL) slowly at -78 C. After addition, the mixture was warmed to 25 C, and stirred at this temperature for 16 hrs. The resulting mixture was diluted with EA (1 L), washed with water (400 mL) three times and brine (500 mL). The separated organic phase was dried over Na2SO4, filtered and concentrated in vacuo to give N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (180.0 g, Compound lf) as a yellow solid and used for next step without further purification. MS
obsd. (ESI ) [(M+H)+]: 339.1.
Step 2: Preparation of N4-benzy1-6-chloro-2-propylsulfanyl-pyrimidine-4,5-diamine (Compound 1g) CI
, I
H
lg
- 109 -To a solution of N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (180 g, Compound 11) and HOAc (319 g, 5.31 mol) in THF(3.0 L) was added Zn (174 g, 2.66 mol) slowly at 25 C. After the addition, the mixture was stirred at 25 C for 16 hrs. The reaction was filtered and the filtrate was basified with saturated aq. NaHCO3 (800 mL), extracted with EA (400 mL) three times, dried over Na2SO4 and concentrated in vacuo.
The residue was purified by silica gel chromatography to give N4-benzy1-6-chloro-2-propylsulfanyl-pyrimidine-4,5-diamine (125 g, Compound 1g) as a brown solid.
MS obsd.
(ESL') [(M+H)+]: 309.1.
Step 3: Preparation of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (Compound 1h) CI
H
NJN
I 1:) SN N
*
th To a solution of N-benzy1-6-chloro-2-(propylsulfanyl)pyrimidine-4,5-diamine (72.0 g, 233.1 mmol, Compound 1g) and CDI (75.2 g, 233.1 mmol) in THF(800mL) was stirred at 80 C for 16 hrs. The resulting mixture was diluted with EA (400 mL), washed with water (200 mL) twice and brine (200 mL). The separated organic layer was dried over Na2SO4, concentrated in vacuo. The residue was washed with MTBE (200 mL) to give 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (58.0 g, Compound 1h) as a white solid and was used in next step without further purification. MS obsd. (ESI ) [(M+H)+]: 335.1.
Step 4: Preparation of 9-benzy1-6-[(4-methoxyphenyl)methylamino]-2-propylsulfanyl-7H-purin-8-one (Compound li)
The residue was purified by silica gel chromatography to give N4-benzy1-6-chloro-2-propylsulfanyl-pyrimidine-4,5-diamine (125 g, Compound 1g) as a brown solid.
MS obsd.
(ESL') [(M+H)+]: 309.1.
Step 3: Preparation of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (Compound 1h) CI
H
NJN
I 1:) SN N
*
th To a solution of N-benzy1-6-chloro-2-(propylsulfanyl)pyrimidine-4,5-diamine (72.0 g, 233.1 mmol, Compound 1g) and CDI (75.2 g, 233.1 mmol) in THF(800mL) was stirred at 80 C for 16 hrs. The resulting mixture was diluted with EA (400 mL), washed with water (200 mL) twice and brine (200 mL). The separated organic layer was dried over Na2SO4, concentrated in vacuo. The residue was washed with MTBE (200 mL) to give 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (58.0 g, Compound 1h) as a white solid and was used in next step without further purification. MS obsd. (ESI ) [(M+H)+]: 335.1.
Step 4: Preparation of 9-benzy1-6-[(4-methoxyphenyl)methylamino]-2-propylsulfanyl-7H-purin-8-one (Compound li)
- 110 -NHPMB
H
NN
I
SIN N
*
li A solution of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (58.0 g, Compound 1h) and PMBNH2 (54.7 g, 398.42 mmol) in n-BuOH (600 mL) was stirred at 120 C for 20 hrs. The reaction was concentrated and the residue was washed with MTBE
(400 mL) to give 9-benzy1-6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-7H-purin-8-one (75 g, Compound li) as a white solid and was used in next step without further purification. MS obsd. (ESL') [(M+H)+]: 436.2.
Step 5: Preparation of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c) NJCNH
I 'C3' SN N
#
lc 9-Benzy1-6-[(4-methoxyphenyOmethylamino]-2-propylsulfanyl-7H-purin-8-one (87.0 g, Compound li) in TFA (200 mL) was stirred at 80 C for 16 hrs. The resulting reaction mixture was concentrated, basified with saturated aq. NaHCO3 (600 mL). The resulting precipitate was collected by filtration and washed with (PE/DCM =
2:1, 400mL) to give 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (38.0 g, Compound 1c) as a white solid. MS obsd. (ESL') [(M+H)+]: 316.1.
Step 6: Preparation of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d) N
I
S)N N
id To a solution of m-CPBA(22.98 g, 113.2 mmol) in THF (50 mL) was added dropwise to a suspension of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (35.0 g, compound 1c) in THF (200 mL) at 0 C. After the addition, the reaction mixture was stirred at 25 C for 0.5 hr. The mixture was filtered and washed with MeCN
(400 mL), MTBE (500 mL) to give 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (35.1 g, Compound 1d) as a white solid, which was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]: 332.1.
Step 7: Preparation of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) ON\
H N=S N N
le To a solution of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (34.0 g, Compound 1d) in Eaton's reagent (170.0 mL, 7.5 wt. % in methanesulphonic acid) was added NaN3 (15.34 g, 253.97 mmol) at 60 C slowly. Then the mixture was stirred at 60 C
for 30 mins. The resulting reaction mixture was cooled to 25 C, poured into ice cold NH3H20 (500 mL, 1 mol/L), extracted with n-BuOH (100 mL) four times and concentrated in vacuo. The residue was purified byprep-HPLC to give 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (10 g, Compound le). 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.65 (br. s., 1H), 7.26-7.37 (m, 5H), 6.98 (br. s., 2H), 4.97 (s, 2H), 4.02 (s, 1H), 3.33 (t, J= 7.53 Hz, 2H), 1.55-1.74 (m, 2H), 0.92 (t, J=7.53 Hz, 3H). MS
obsd. (ESE') [(M H)+]: 347.
Example 2 6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NH2 ...-N
T \
N----N
o 1 1 µNs' ..--N
......õ,..õ......õ, µN N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-(2-methoxyethyl)-N-methyl-carbamoyl chloride (Intermediate AB) instead ofN-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (120 mg, Example 2) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
7.27-7.39 (m, 5H), 6.89 (br. s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br. dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J= 8Hz, 3H). MS obsd. (Esr) [(M+H)+]: 462.
Separation of compound of Example 2 by chiral HPLC afforded Example 2-A
(faster eluting, 33 mg) and Example 2-B (slower eluting, 46 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak OJ-3 column.
Example 2-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.89 (br.
s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br.
dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J
= 8Hz, 3H). MS obsd. (ESL') [(M+H)+]: 462.
Example 2-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.89 (br.
s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br.
dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J
= 8Hz, 3H). MS obsd. (ESL') [(M+H)+]: 462.
Example 3 6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 r----1 N'\._.....N
9µ j 0 .Sµx -N N
NH
AP
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-propyl-carbamoyl chloride (Intermediate AC) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (51 mg, Example 3) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.85 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. d, J= 8.0 Hz, 1H), 3.13-3.54 (m, 4H), 1.46-1.72 (m, 4H), 1.30-1.39 (m, 1H), 1.00-1.26 (m, 6H), 0.81-0.95 (m, 5H), 0.73 (t, J= 8 Hz, 1H). MS
obsd. (ESL') [(M+H)+]: 474.
Example 4 6-Amino-9-benzy1-7-14-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one N
NJ
//Sµµr\JH N "
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using (1,4'-bipiperidine)-1'-carbonyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one (55 mg, Example 4) was obtained as a white powder.
- 114 -11-1 NMR (400 MHz, DMSO-d6) 6 ppm: 7.39 - 7.27 (m, 5H), 6.97 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. s., 2H), 3.85 (d, J= 12.5 Hz, 1H), 3.43 - 3.15 (m, 3H), 2.96 (t, J=
12.3 Hz, 2H), 2.56 (m, 4H), 1.83 (m, 1H), 1.79 - 1.54 (m, 4H), 1.50 (br. s., 4H), 1.45 -1.33 (m, 3H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 541.2.
Example 5 6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyDpurine-7-carboxamide 0 rj \---Nj----N
µµS
N 1\1 ---NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (Intermediate AD) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (34 mg, Example 5) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.39 -7.28 (m, 5H), 6.89 (br. s., 1H), 6.74 (br. s., 1H), 4.99 (s, 2H), 4.17 (d, J= 8.1 Hz, 1H), 3.67 (br. s., 2H), 3.63 - 3.51 (m, 2H), 3.50 - 3.34 (m, 4H), 3.29 (s, 1H), 3.11 (s, 2H), 1.73- 1.59 (m, 2H), 1.23 - 1.07 (m, 3H), 0.93 (t, J= 7.5 Hz, 3H). MS obsd. (Esr) [(M+H)+]:
476.3.
Example 6 6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyDpurine-7-carboxamide 0 ri-Nj\.___N
9µ 0 //Sµµ N N
NH
At The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-butyl-N-ethyl-carbamoyl chloride (Intermediate AE) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-butyl-N-ethy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (51 mg, Example 6) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.85 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. d, J= 8.0 Hz, 1H), 3.13-3.54 (m, 4H), 1.46-1.72 (m, 4H), 1.30-1.39 (m, 1H), 1.00-1.26 (m, 6H), 0.81-0.95 (m, 5H), 0.73 (t, J= 8 Hz, 1H). MS obsd.
(ESL') [(M+H)+]: 474.
Example 7 6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NH2 .......N
Ni\li \\
9µ )=0 Sµµ N N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (Intermediate AF) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (35 mg, Example 7) was obtained as a white powder. 'H NMR (400 MHz, DMSO-d6) 6 ppm:
7.40 - 7.28 (m, 5H), 6.89 (br. s., 1H), 6.75 (br. s., 1H), 5.00 (d, J= 5.5 Hz, 2H), 4.24 - 4.16 (m, 1H), 3.77 (br. s., 1H), 3.67 (br. s., 1H), 3.62 - 3.53 (m, 1H), 3.42 - 3.27 (m, 5H), 3.23 -3.02 (m, 3H), 1.66-1.38 (m, 4H), 0.96 - 0.70 (m, 6H). MS obsd. (ESL') [(M+H)+]: 490.5.
Example 8 6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NJ.---9µ j 0 SN N N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using bis(2-methoxyethyl)carbamic chloride (Intermediate AG) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (35 mg, Example 8) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.40 - 7.28 (m, 5H), 6.83 (br. s., 2H), 4.99 (s, 2H), 3.71 (br. s., 3H), 3.52 - 3.27 (m, 11H), 3.09 (s, 3H), 1.73- 1.59 (m, 2H), 0.93 (t, J= 7.5 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 506.
Example 9 6-Amino-7-(azetidine-1-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one NN
OµN j I
S' N' N
NH
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using azetidine-l-carbonyl chloride (Intermediate AH) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-7-(azetidine-1-carb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one (120 mg, Example 9) was obtained as a white powder.
1HNMR (400 MHz, DMSO-d6) 6 ppm: 7.02 - 7.43 (m, 7H), 4.99 (s, 2H), 4.31 (t, J=
7.65 Hz, 2H), 4.08 - 4.23 (m, 3H), 3.34 - 3.41 (m, 2H), 2.28 (m, 2H), 1.56 - 1.73 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 430.
Example 10 6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2 ......1\1 / \
N----N
µµS N
/..\,=-=' µµ N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 10 using N-isopropyl-N-methyl-carbamoyl chloride (Intermediate Al) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (97 mg, Example 10) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.87 (br. s., 2H), 4.99 (s, 2H), 4.38-4.45 (m, 1H), 4.09-4.21 (m, 1H), 3.29-3.43 (m, 2H), 2.89-2.95 (m, 3H), 1.58-1.73 (m, 2H), 1.21 (br d, J= 8 Hz, 6H), 0.93 (t, J= 8 Hz, 3H). MS obsd.
(ESL') [(M+H)+]: 446.
Example 11 6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one 0 r--\ N---NH2 ...... N \..... j N----N
0µµ I
S' N "
NH
*
ii The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 4-methylpiperazine-1-carbonyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-(propylsulfonimidoyl)purin-8-one (59.5 mg, Example 11) was obtained as a yellow solid.
1H NMR (400 MHz, DMSO-d6) gppm: 7.39 - 7.31 (m, 5H), 6.99 (s, 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.58 - 3.49 (m, 6H), 2.42 (m, 4H), 2.22 (s, 3H), 1.66 - 1.61 (m, 2H), 0.95 -0.91 (t, J= 7.2 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 473.
Example 12 6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide /
0 ri NH2 ...-N
J \
N-----N
o1Io "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-(3-methoxypropy1)-N-methyl-carbamoyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (92.2 mg, Example 12) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.23 - 7.45 (m, 5H), 6.94 (s., 2H), 4.93-5.08 (m, 2H), 4.19 (s, 1H), 3.30 - 3.62 (m, 6H), 3.25 (s, 3H), 3.02 - 3.10 (m, 3H), 1.74 - 1.90 (m, 2H), 1.55 - 1.77 (m, 2H), 0.98 - 0.82 (m, 3H). MS obsd.
(ESL') [(M+H)+]: 476.3.
Example 13 6-Amino-9-benzyl-N-isobutyl-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2 ...-N
\
N
N.---µµ ...---N
//Sµ\NH N
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-isobutyl-N-methyl-carbamoyl chloride (Intermediate AL) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-isobutyl-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (64 mg, Example 13) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.40 (m, 5H), 6.89 (br. s., 2H), 5.00 (s, 2H), 4.16 (br. s., 1H), 3.25-3.44 (m, 4H), 3.07 (s, 2H), 3.03 (s, 1H), 1.87-2.09 (m, 1H), 1.57-1.74 (m, 2H), 0.75-0.99 (m, 9H). MS obsd. (ESL') [(M+H)+]: 460.
Example 14 Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]acetate 0---j 0 r40 NH2 ..-N
1 \
N.......N
ON\ IIo "
\µ N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl 2-((chlorocarbonyl)(methyl)amino)acetate (Intermediate AP) instead ofN-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl 24[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] acetate (38 mg, Example 14) was obtained as a light yellow powder. 1H NMR (400MHz, DMSO-d6) 6 ppm:
7.41 -7.27 (m, 5H), 6.82 (br. s., 1H), 5.04 - 4.95 (m, 2H), 4.35 (br. s., 1H), 4.28 (br. s., 1H), 4.23 - 4.16 (m, 2H), 4.08 (q, J= 7.2 Hz, 1H), 3.43 - 3.28 (m, 3H), 3.15 (s, 2H), 3.08 (s, 1H), 1.71 - 1.58 (m, 2H), 1.24 (t, J= 7.0 Hz, 2H), 1.12 (t, J= 7.0 Hz, 1H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 490.
Example 15 Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate 0 r r ,..... 0 NNII \
(:)µµ I
µ\ N "
NH
ilt 15 The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl 3-((chlorocarbonyl)(methyl)amino)propanoate instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate (35 mg, Example 15) was obtained as a white powder. 'H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.26 (m, 5H), 6.93 (br. s., 2H), 4.99 (s, 2H), 4.16 (s, 1H), 4.08 (q, J= 7.1 Hz, 1H), 3.99 (d, J= 7.0 Hz, 1H), 3.67 (br. s., 2H), 3.40 - 3.29 (m, 2H), 3.08 (s, 2H), 2.99 (s, 1H), 2.71 (t, J= 6.4 Hz, 2H), 1.74- 1.56 (m, 2H), 1.27- 1.05 (m, 3H), 0.93 (t, J= 7.5 Hz, 3H). MS
obsd.
(ES[) [(M+H)+]: 504.
Example 16 tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate 0 \/----ry,0 \
N
N.--µN -----N1 N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl 3-[chlorocarbonyl(methyl)amino]propanoate (Intermediate AR) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate (60 mg, Example 16) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
7.41 - 7.27 (m, 5H), 6.93 (br. s., 2H), 4.99 (s, 2H), 4.15 (s, 1H), 3.64 (br.
s., 2H), 3.51 -3.33 (m, 2H), 3.08 (s, 2H), 2.98 (s, 1H), 2.62 (t, J= 6.9 Hz, 2H), 1.71 - 1.57 (m, 2H), 1.41 (s, 6H), 1.34 (s, 3H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ES[) [(M+H)+]: 532.
Example 17 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate \
Nj\____N
oNN
SNµ N
NH N
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (2S)-2-[chlorocarbonyl(methyl)amino]propanoate (Intermediate AS) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate (34.1 mg, Example 17) was obtained as a yellow solid. 1H NMR (300 MHz, DMSO-d6) 6 ppm:
7.22 - 7.49 (m, 5 H), 6.78 (br. s., 2H), 4.93 - 5.08 (m, 2H), 4.75 (br. s., 1H), 3.96 - 4.29 (m, 3H), 3.30 - 3.46 (m, 2H), 3.09 (s, 2H), 2.93 (br. s., 1H), 1.55- 1.77 (m, 2H), 1.48 (d, J=
7.16 Hz, 3H), 1.09 - 1.29 (m, 3H), 0.94 (t, J = 7.44 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
504.2.
Example 18 tert-Butyl (2S)-2- I [6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] -4-methyl-pe ntano ate _........4)--\
Nj\õ.....N
.
oxµ 0 "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AT) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (22 mg, Example 18) was obtained as a white solid.
1H NMR (400MHz, DMSO-d6) 6 ppm: 7.42 - 7.27 (m, 5H), 6.78 (br. s., 2H), 5.05 -4.96 (m, 2H), 4.78 (br. s., 1H), 4.33 (br. s., 1H), 3.51 - 3.37 (m, 2H), 3.01 (s, 3H), 1.75 - 1.54 (m, 4H), 1.44 (s, 8H), 1.33- 1.11 (m, 2H), 0.99 - 0.82 (m, 9H). MS obsd.
(ESL') [(M+H)+]:
574.3.
Example 19 Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]-4-methyl-pentanoate o 0 \
Nj\,....N
0 I I .
µNS N .....õ,........õ,/, µN
NH
N
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AU) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (43 mg, Example 19) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.27 (m, 5H), 6.75 (br. s., 2H), 5.05 - 4.94 (m, 3H), 4.88 (br. s., 1H), 4.19 (br. s., 1H), 3.43 - 3.34 (m, 2H), 3.01 (s, 3H), 1.91 (br. s., 1H), 1.77 - 1.56 (m, 4H), 1.25 - 1.16 (m, 6H), 0.99 - 0.83 (m, 9H).
MS obsd. (ESC') [(M+H)+]: 560.3.
Example 20 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate 0¨i o-----40 NH2 ...-N
\
N
N--9µ I
S\\ N "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-methyl-butanoate (Intermediate AV) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-5 amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate (51.5 mg, Example 20) was obtained as a white powder. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.23 - 7.51 (m, 5H), 6.76 (br. s., 2H), 5.01 (br. s., 2H), 4.42 (br. s., 1H), 3.97 - 4.26 (m, 3H), 3.34 - 3.45 (m, 2H), 3.12 (br. s., 3H), 2.24 (br.
s., 1H), 1.65 (br. s., 2H), 1.13 - 1.29 (m, 3H), 0.88 - 1.10 (m, 9H). MS obsd. (ESI+) [M+H ]: 532.2.
10 Example 21 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate J
0 ......:40 NH2 ..-N
\
N
N--9µ I
Sµµ N "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 15 using ethyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AW) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (17.3 mg, Example 21) was obtained as a white powder. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.26 - 7.45 (m, 5H), 6.73 (br. s., 2H), 4.91 - 5.09 (m, 3H), 4.06 -4.25 (m, 3H), 3.34 - 3.45 (m, 2H), 3.04 (br. s., 3H), 1.93 (br. s., 1H), 1.54 -1.78 (m, 4H), 1.22 (t, J= 7.09 Hz, 3H), 0.77 - 1.01 (m, 9H). MS obsd. (ES[) [(M+H)+]: 546.3.
Example 22 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]-3-phenyl-propanoate NH2 ...-N
\
N\ õ...... N
N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (25)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AX) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (30 mg, Example 22) was obtained as a white powder. 1H NMR
(400MHz, DMSO-d6) 6 ppm: 7.42 - 7.16 (m, 10H), 4.97 (s, 3H), 4.19 (q, J= 7.1 Hz, 2H), 3.35 -3.15 (m, 6H), 3.10 - 2.90 (m, 3H), 1.71 - 1.46 (m, 2H), 1.28- 1.18 (m, 4H), 0.97 -0.85 (m, 3H). MS obsd. (ES[) [(M+H)+]: 580.
Example 23 Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate 4Ik 0----( \
Nj\,....N
oNN .
\µ N
NH
"
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using isopropyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AY) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (22 mg, Example 23) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.35 - 7.01 (m, 10H), 5.02-4.89 (m, 3H), 3.37-3.17 (m, 3H), 3.02 - 3.09 (m, 3H), 3.10 -2.90 (m, 3H), 1.66 - 1.62 (m, 2H), 1.22 -1.11 (m, 8H), 0.92 (t, J= 7.4 Hz, 3H). MS obsd. (ESI+) [(M+H)+]: 594.
Example 24 tert-Butyl (2S)-2-[16-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate 4Ik 0----k-N--- NI \
% 1.
-N N
NH
4111\
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AZ) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl (2,5)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (34 mg, Example 24) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.42 - 7.16 (m, 10H), 5.03 - 4.90 (m, 3H), 3.68 - 3.24 (m, 5H), 3.24 - 3.09 (m, 2H), 3.01 (s, 3H), 1.68 - 1.57 (m, 2H), 1.43 (s, 9H), 0.99 - 0.85 (m, 3H). MS obsd. (ESL') [(M+H)+]: 608.3.
Example 25 N- 12- [Acetyl(methyl)amino] ethyl] -6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide \ N
NH2 ...-N
1 \
N'N
ID\ I
S
-----N
µ N
...õ..."--.........õ., µµ
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N- [2-[acetyl(methyl)amino]ethyl]-N-methyl-carbamoyl chloride (Intermediate BA) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). N-[2-15 [Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (26.1 mg, Example 25) was obtained as a white powder.1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.27 (m, 5H), 7.02 (br, 2H), 5.04 - 4.97 (m, 2H), 4.19 - 4.13 (m, 1H), 3.57 (d, J= 5.5 Hz, 2H), 3.49 - 3.34 (m, 2H), 3.14 (s, 1H), 3.12 - 3.02 (m, 4H), 2.86 (d, J= 7.5 Hz, 2H), 2.69 - 2.64 (m, 1H), 2.05 (s, 1H), 20 1.99 (s, 1H), 1.91 - 1.83 (m, 1H), 1.70 - 1.59 (m, 2H), 0.97 - 0.90 (m, 3H). MS obsd.
(ESI ) [(M+H)+]: 503.2.
Example 26 Methyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino] ethyl] -N-methyl-c arbam ate \ N40 0 r---1 NH2 ......N
\
N
N).--% I
Sµµr\jH N "
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using methyl N42-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (Intermediate BB) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (65 mg, Example 26) was obtained as a yellow solid. 1H NMR (400 MHz, CDC13) 6 ppm: 7.29 - 7.49 (m, 5H), 5.63 - 5.92 (m, 2H), 5.03 -5.17 (m, 2H), 3.43 - 3.69 (m, 8H), 3.13 - 3.27 (m, 3H), 2.96 - 3.05 (m, 2H), 2.72 (br. s., 1H), 1.05 (t, J= 7.40 Hz, 3H), 1.87 (dd, J= 14.12, 6.96 Hz, 2H). MS obsd.
(ES[) [(M+H)+]: 519.2.
Example 27 tert-Butyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino] ethyl] -N-methyl-c arbam ate O---k \ N40 0 ri \
N
N
ONN 1 )=C) µ N "
'NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl N- [2-[chlorocarbonyl(methyl)amino]ethyl]-N-methyl-carbamate (Intermediate BC) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (32 mg, Example 27) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.26 (m, 5H), 6.89 (br. s., 2H), 4.99 (d, J= 5.0 Hz, 2H), 4.16 (s, 1H), 3.55 (br. s., 2H), 3.48 - 3.34 (m, 2H), 3.10 (s, 2H), 3.07 (s, 1H), 2.86 (d, J= 12.8 Hz, 2H), 2.74 (d, J= 9.5 Hz, 1H), 2.70 - 2.60 (m, 1H), 1.72 - 1.54 (m, 2H), 1.39 (s, 6H), 1.23 (s, 2H), 1.13 (s, 2H), 0.93 (t, J= 7.4 Hz, 3H). MS
obsd. (Esr) [(M+H)+]: 562.
Example 28 Ethyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino] ethyl] -N-methyl-carbamate \ N40 0 ri NH2 ...-N
N---Nil \
oµµ j 0 -N N
*15 "NH 28 The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl N42-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (Intermediate BD) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl N-[2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (87 mg, Example 28) was obtained as a yellow solid.1H
NMR (400 MHz, CDC13) 6 ppm: 7.29 - 7.53 (m, 5H), 5.65 - 5.90 (m, 2H), 5.02 -5.14 (m, 2H), 3.38 - 4.21 (m, 9H), 3.14 - 3.26 (m, 3H), 3.00 (br. s., 2H), 2.73 (s, 1H), 1.76 - 1.99 (m, 2H), 1.22 - 1.31 (m, 3H), 1.05 (s, 3H). MS obsd. (ESE') [(M+H)+]: 533.2.
Example 29 2-116-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyli-methyl-amino] ethyl N-butyl-N-methyl-carbamate N
NH2 ...-N
j \
µµ N S N
............õ µµ
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (Intermediate BE) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl N-butyl-N-methyl-carbamate (19 mg, Compound 29) was obtained as yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.25 - 7.48 (m, 5H), 6.96 (br. s., 2H), 4.99 (s, 2H), 4.06 - 4.36 (m, 3H), 3.59 - 3.83 (m, 1H), 3.33 - 3.49 (m, 3H), 3.07 - 3.21 (m, 4H), 2.79 (s, 2H), 1.65 (br. s., 2H), 1.05 - 1.47 (m, 6H), 0.93 (t, J= 7.40 Hz, 3H), 0.70 - 0.87 (m, 3H). MS
obsd. (ESE) [(M+H)+]: 561.2.
Example 30 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl pyrrolidine-l-carboxylate Cs N----0 rj NH2 ....- N
\
Nj\,....N
O I
N\
/\Sµµ " "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 5 using 2-[chlorocarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (Intermediate BF) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl pyrrolidine-l-carboxylate (10.0 mg, Example 30) was obtained as a yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.26 - 7.41 (m, 5H), 6.96 (br.s., 2H), 4.99 (s, 2H), 4.01 -4.35 (m, 4H), 10 3.29 - 3.47 (m, 3H), 3.23 (br. s., 3H), 3.03 - 3.17 (m, 4H), 1.52 - 1.84 (m, 6H), 0.90 ¨0.96 (m, 3H). MS obsd. (ESI+) [(M+H)+]: 545.2.
Example 31 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyli-methyl-aminoiethyl N-methyl-N-propyl-carbamate \ NX
0 ri NH2 ...-N
\
NN
9µ 0 N
S\µ N
NH
=
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl N-methyl-N-propyl-carbamate (Intermediate BG) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-methyl-N-propyl-carbamate (3.7 mg, Example 31) was obtained as a yellow solid. 1H NMR
(400 MHz, CD30D) 6 ppm: 7.22 - 7.48 (m, 5H), 5.09 - 5.22 (m, 4H), 4.55 (s, 2H), 3.38 - 3.57 (m, 4H), 3.13 (s, 3H), 1.61 - 1.85 (m, 4H), 1.22- 1.41 (m, 3H), 0.88 - 1.13 (m, 6H). MS
obsd. (ESI+) [(M+H)+]: 547.2.
Example 32 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N,N-diethylcarbamate NJN
0 rj NH2 ....-N
T \
Nj\_....N
9µ )=0 "
\µ N
NH
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl /V,N-diethylcarbamate (Intermediate BH) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl N,N-diethylcarbamate (21.7 mg, Example 32) was obtained as yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.25 - 7.41 (m, 5H), 6.96 (br. s., 2H), 4.99 (s, 2H), 4.08 - 4.36 (m, 3H), 3.70 (br, 1H), 3.33 - 3.46 (m, 3H), 3.01 - 3.24 (m, 7H), 1.55 - 1.74 (m, 2H), 0.86 -1.05 (m, 9H). MS obsd. (ESL') [(M+H)+]: 547.2.
Example 33 2-116-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl ethyl carbonate 0-i 0 ri NH2 ...-N
NN1 \
N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl ethyl carbonate (Intermediate BI) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl ethyl carbonate (46 mg, Example 33) was obtained as yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
0.82 -0.99 (m, 3H), 1.02 - 1.28 (m, 3H), 1.56 - 1.76 (m, 2H), 3.05 - 3.18 (m, 3H), 3.35 - 3.48 (m, 3H), 3.73 (t, J= 5.08 Hz, 2H), 4.08 - 4.27 (m, 3H), 4.37 (br. s., 1H), 5.00 (s, 2H), 6.76 -7.11 (m, 2H), 7.22 - 7.45 (m, 5H). MS obsd. (ESE') [(M+H)+]: 520.
Example 34-A and Example 34-B
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-IS(S)-propylsulfonimidoyl]purine-7-carboxamide and 6-amino-N-buty1-9-1(4-chlorophenyllmethyli-N-methyl-8-oxo-2-1S(S)-propylsulfonimidoylipurine-7-carboxamide r r NitoyN NH20N
N\...õ, N
NJ---N
cNIN s ,, )=0 H NNN .0L I )=0 ' N N , S N N
I-1 NV 0' = CI . CI
Step 1: Preparation of 4-amino-3-[(4-chlorophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 34a) N.........H
N
1 ) _____________________________________ 0 34a Compound 34a was prepared in analogy to Example 1, Method A, Step 1 by using 4-chlorobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-chlorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (8.0 g, Compound 34a) was obtained as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 249.
Step 2: Preparation of 6-amino-9-[(4-chlorophenyl)methyl]-2-sulfany1-7H-purin-one (Compound 34b) H
N-----"N
1 ) __ 0 H S-1\1.-----N
34h Compound 34b was prepared in analogy to Example 1, Method A, Step 2 by using 4-Amino-3-[(4-chlorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 34a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la).
6-Amino-9-[(4-chlorophenyl)methy1]-2-sulfany1-7H-purin-8-one (6.4 g, Compound 34b) was obtained as a yellow solid and was used for the next step without further purification.
MS obsd. (ESL') [(M+H)+]: 308.
Step 3: Preparation of 6-amino-9-[(4-chlorophenyhmethy1]-2-propylsulfany1-7H-purin-8-one (Compound 34c) H
N.------N
1 > __ 0 =SNN
= CI
34c Compound 34c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-chlorophenyOmethy1]-2-sulfany1-7H-purin-8-one (Compound 34b) instead of 6-amino-9-phenylmethy1-2-sulfany1-7H-purin-8-one (Compound lb). 6-Amino-9-[(4-chlorophenyOmethy1]-2-propylsulfany1-7H-purin-8-one (800 mg, Compound 34c) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 350.
Step 4: Preparation of 6-amino-9-[(4-chlorophenyhmethy1]-2-propylsulfiny1-7H-purin-8-one (Compound 34d) H
N-------N
1 ) __ 0 SNN
II
34d Compound 34d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-propylsulfany1-7H-purin-8-one (Compound 34c) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-9-[(4-chlorophenyl)methy1]-2-propylsulfiny1-7H-purin-8-one (150 mg, Compound 34d) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 366.
Step 5: Preparation of 6-amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (compound 34e), 6-amino-9-[(4-chlorophenyl)methy1]-2-1S(S)-propylsulfonimidoy1)-7H-purin-8-one and 6-amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-propylsulfonimidoy1)-7H-purin-8-one (Compound 34e-A and Compound 34e-B) H
N----N
I-1 N\\ I > __ 0=S NN
H = CI
34e H
N-----N
NN H N 1 >
--- _______________________________________________________________ 1 0 1 > ___ \\ ,L
\\ 0.-<.. 0=S 's N'---N
H N=S 's N--N
= CI
= CI
34e-A and 34e-B
Compound 34e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-propylsulfiny1-7H-purin-8-one (Compound 34d) instead of 6-amino-9-benzy1-2-(2-propylsulfiny1)-7H-purin-8-one (Compound 1d).
Amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (250 mg, compound 34e) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
10.60 (br. s, 1H), 7.32-7.42 (m, 4H), 6.98 (br. s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.25-3.41 (m, 2H), 1.56-1.68 (m, 2H), 0.91 (t, J= 8 Hz, 3H). MS obsd. (ESL') [(M+1-1) ]:
381.
Separation of compound of Compound 34e by chiral HPLC afforded Compound 34e-A (faster eluting, 110 mg) and Compound 34e-B (slower eluting, 100 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak OJ-3 column.
Compound 34e-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.63 (br. s, 1H), 7.33-7.42 (m, 4H), 6.99 (br. s, 2H), 4.96 (s, 2H), 4.05 (br. s, 1H), 3.26-3.39 (m, 2H), 1.53-1.69 (m, 2H), 0.91 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 381.
Compound 34e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.63 (br. s, 1H), 7.33-7.42 (m, 4H), 6.99 (br. s, 2H), 4.96 (s, 2H), 4.05 (br. s, 1H), 3.26-3.40 (m, 2H), 1.54-1.69 (m, 2H), 0.91 (t, J= 7.5 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 381.
Step 6: 6-Amino-N-buty1-9-[(4-chlorophenyl)methyll-N-methy1-8-oxo-2-1S(S)-propylsulfonimidoyl]purine-7-carboxamide and 6-amino-N-buty1-9-[(4-chlorophenyl)methyll-N-methy1-8-oxo-2-1S(S)-propylsulfonimidoyl]purine-7-carboxamide (Example 34-A and Example 34-B) r NI-I21/4j, N NH2L*N
N N N.....,..N
-' oNN ,, 0 I-1 NµN s, I )=0 ,S ' N N ,S
I-1 NV 0' = CI IIIP CI
Example 34-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 34e-A and N-butyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound 1e) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 34-A (160 mg): 'H NMR (400 MHz, DMSO-d6) 6 ppm: 7.37-7.45 (m, 4H), 6.91 (br. s., 2H), 4.99 (s, 2H), 4.17 (s, 1H), 3.28-3.40 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 1.49-1.70 (m, 4H), 1.15-1.37 (m, 2H), 0.89-0.94 (m, 5H), 0.76 (t, J= 8 Hz, 1H). MS
obsd. (ESL') [(M+H)+]: 494.
Example 34-B (167 mg) was prepared in analogy to Example 34-A by using Compound 34e-B instead of Compound 34e-A.
Example 34-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.36-7.45 (m, 4H), 6.91 (br.
s., 2H), 4.99 (s, 2H), 4.17 (s, 1H), 3.28-3.41 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 1.50-1.71 (m, 4H), 1.15-1.37 (m, 2H), 0.89-0.94 (m, 5H), 0.76 (t, J= 7.4 Hz, 1H). MS obsd.
(ESE) [(M+H)+]: 494.
Example 35 6-Amino-9-[(4-chlorophenyl)methyli-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2uyN
N
N--oNN 0 S N N
I-INV
. CI
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 20 using 6-amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound 34e) and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (60 mg, Example 35) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.40 (s, 4H), 6.91 (br s, 2H), 4.99 (s, 2H), 4.16 (s, 1H), 3.34-3.44 (m, 4H), 3.05 (s, 2H), 3.01 (s, 1H), 1.58-1.67 (m, 2H), 1.18 (t, J= 8.0 Hz, 3H), 0.92 (t, J= 8.0 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 466.
Example 36-A and Example 36-B
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy11-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide N--N N--'N
I-1 NN, .s,I o s, Nµ 0 ,S N N S ' NJ N
0' . I-1 NV
Step 1: Preparation of 6-chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (Compound 36a) CI
Ni S)N 1 N H
36a Compound 36a was prepared in analogy to Example 1, Method B, Step 1 by using p-tolylmethylamine instead of phenylmethanamine. 6-Chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (3.9 g, Compound 36a) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 353.
STEP 2: PREPARATION OF 6-CHLOR0-2-PROPYLSULFANYL-N4-(P-TOLYLMETHYL)PYRIMIDINE-4,5-DIAMINE (COMPOUND 36B) CI
S(I
N NH
36b Compound 36b was prepared in analogy to Example 1, Method B, Step 2 by using 6-chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (Compound 36a) instead of N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (Compound if). 6-Chloro-2-propylsulfanyl-N4-(p-tolylmethyl)pyrimidine-4,5-diamine (2.2 g, Compound 36b) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]:
323.
STEP 3: PREPARATION OF 6-CHLOR0-2-PROPYLSULFANYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36C) CI
H
NJN
I
SIN N
36c Compound 36c was prepared in analogy to Example 1, Method B, Step 3 by using 6-chloro-2-propylsulfanyl-N4-(p-tolylmethyl)pyrimidine-4,5-diamine (Compound 36b) instead of N-benzy1-6-chloro-2-(propylsulfanyl)pyrimidine-4,5-diamine (Compound 1g).
6-Chloro-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (2.2 g, Compound 36c) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 349.
STEP 4: PREPARATION OF 6-1-(4-METHOXYPHENYL)METHYLAMIN0]-2-PROPYLSULFANYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36D) NHPMB
H
NN
I
N
36d Compound 36d was prepared in analogy to Example 1, Method B, Step 4, by using 6-chloro-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36c) instead of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (Compound 1h). 6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (2.0 g, Compound 36d) was obtained as a white solid. MS obsd. (ESE) [(M+H)+]: 450.
STEP 5: PREPARATION OF 6-AMINO-2-PROPYLSULFAIVYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36E) N\_ S)N N
36e Compound 36e was prepared in analogy to Example 1, Method B, Step 5 by using 6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36d) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound ii). 6-amino-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (1.0 g, Compound 36e) was obtained as a white solid. MS obsd. (ESE) [(M+H)+]: 330.
STEP 6: PREPARATION OF 6-AMINO-2-PROPYLSULFINYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36F) H
I
N
36f Compound 361 was prepared in analogy to Example 1, Method B, Step 6 by using 6-amino-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36e) instead of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound 1c). 6-amino-2-propylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (220 mg, Compound 36f) was obtained as a white solid MS obsd. (ESL') [(M+H)+]: 345.
STEP 7: PREPARATION OF 6-AMINO-2-(PROPYLSULFONIMIDOYL)-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36G) NN
0 >
\\
H N=S N
36g Compound 36g was prepared in analogy to Example 1, Method B, Step 7 by using 6-amino-2-propylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 361) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (127 mg, Compound 36g) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.67 (br. s., 1H), 7.23 (d, J= 8.0 Hz, 2H), 7.13 (d, J= 8.0 Hz, 2H), 6.98 (br. s., 2H), 4.91 (s, 2H), 4.05 (s, 1H), 3.34-3.27 (m, 2H), 2.26 (s, 3H), 1.67-1.62 (m, 2H), 0.92 (t, J= 8.0 Hz, 3H).
MS obsd.
(ESL') [(M+H)+]: 361.
Separation of compound 36g by chiral HPLC afforded compound 36g-A (faster eluting, 50 mg) and compound 36g-B (slower eluting, 49 mg) as white solid with 30%
isopropanol (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 36g-A: 111 NMR: (400 MHz, DMSO-d6) 6 ppm: 10.51 (s, 1 H), 7.22 (d, J=
8.0 Hz, 2H), 7.12 (d, J= 8.0 Hz, 2H), 7.00 (s, 2 H), 4.91 (s, 2H), 4.03 (s, 1H), 3.35 - 3.31 (m, 2H), 2.26 (s, 3H), 1.70 - 1.58 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd.
(ESI ) [(M+H)+]: 361.
Compound 36g-B: 111 NMR: (400 MHz, DMSO-d6) 6 ppm: 10.54 (s, 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J= 8.0 Hz, 2H), 6.97 (s, 2H), 4.91 (s, 2H), 4.04 (s, 1H), 3.34 - 3.30 (m, 2H), 2.26 (s, 3H), 1.72 - 1.57 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd.
(ESL') [(M+H)+]: 361.
Step 8: Preparation of 6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-methyl-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide (Example 36-A and Example 36-B) , , NH2L'yN NH-'N
NCN
H Nµµ ,µ I 0 (3µ L
,\S's% N N
I¨IN' 0' 0 *
Example 36-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). Example 36-A (108 mg) was obtained as a white solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.27 (d, J= 8 Hz, 2H), 7.14 (d, J= 8 Hz, 2H), 6.87 (br.
s., 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.33-3.57 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 2.26 (s, 3H), 1.52-1.73 (m, 4H), 0.75-0.97 (m, 6H). MS obsd. (ESL') [(M+H)+]: 460.
Example 36-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (compound le). Example 36-B(125 mg): 1H NMR (400 MHz, DMSO-d6) 5 ppm: 7.27 (d, J= 8 Hz, 2H), 7.14 (d, J= 8 Hz, 2H), 6.87 (br. s., 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.33-3.57 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 2.26 (s, 3H), 1.52-1.73 (m, 4H), 0.75-0.97 (m, 5H). MS
obsd. (ESE') [(M+H)+]: 460.
Example 37-A and Example 37-B
6-Amino-2-1S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one and 6-amino-2-1S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one 0 NDNH2 NI-12()ND
N'\........N
Nj\õ.....N
I-INN\ s, I )=0 cNIN oL 0 ,S ' N N S
0' 0 I-INV
.
Example 37-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A and pyrrolidine-l-carbonyl chloride instead of 6-amino-9-benzy1-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 37-A (390 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.31 -7.11 (m, 4H), 7.04 (s, 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.65 - 3.47 (m, 4H), 3.37 (m, 2H), 2.27 (s, 3H), 1.97 - 1.81 (m, 4H), 1.71 - 1.59 (m, 2H), 0.94 (t, J=
7.4 Hz, 3H). MS obsd. (ESE') [(M+H)+]: 458.2.
Example 37-B (125 mg) was prepared in analogy to Example 37-A by using Compound 36g-B instead of Compound 36g-A. 1H NMR (400 MHz, DMSO-d6) gppm:
7.28 - 7.14 (m, 4H), 7.04 (s, 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.65 - 3.47 (m, 4H), 3.37 (m, 2H), 2.27 (s, 3H), 1.93 - 1.84 (m, 4H), 1.65 - 1.60 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESE') [(M+H)+]: 458.3.
Example 38-A and Example 38-B
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2-1S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2-IS(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide N I-12L'N NH2L*N
N N
FINN\ s, 0 ,S 's N N S = N N
0' . HN
*
Example 38-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A and N-(2-methoxyethyl)-N-methyl-carbamoyl chloride (Intermediate AB) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy0-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 38-A (57.8 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.26 (d, J= 7.6 Hz, 2H), 7.14 (d, J= 7.6 Hz, 2H), 6.89 - 6.78 (m, 2H), 4.95 (s, 2H), 4.18 (s, 1H), 3.62 -3.58 (m, 2H), 3.43 - 3.37 (m, 2H), 3.30 -3.10 (m, 3H), 3.09 - 3.08 (m, 3H), 3.08 - 3.05 (m, 2H), 2.27 (s, 3H), 1.77 - 1.54 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 476.3.
Example 38-B (46.6 mg) was prepared in analogy to Example 38-A by using Compound 36g-B instead of Compound 36g-A. 1H NMR (400 MHz, DMSO-d6) gppm:
7.26 (d, J= 7.6 Hz, 2H), 7.14 (d, J= 7.6 Hz, 2H), 6.89 - 6.78 (m, 2H), 4.95 (s, 2H), 4.18 (s, 1H), 3.62 -3.58 (m, 2H), 3.43 - 3.37 (m, 2H), 3.30 - 3.10 (m, 3H), 3.09 - 3.08 (m, 3H), 3.08 - 3.05 (m, 2H), 2.27 (s, 3H), 1.77 - 1.54 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESL') [(M+H)+]: 476.3.
Example 39 6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide 0 r----NH2 ....... N
\
N
NJ"
9µ I
//Sµµ N N
NH
it The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-methyl-carbamoyl chloride and 6-amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36g) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA) and 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide (141.8 mg, Example 39) was obtained as a light yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.26 (d, J = 7.9 Hz, 2H), 7.15 (d, J =
7.9 Hz, 2H), 6.89 (s, 2H), 4.95 (s, 2H), 4.24 - 4.07 (m, 1H), 3.52 - 3.35 (m, 4H), 3.10 - 2.95 (m, 3H), 2.26 (s, 3H), 1.77 - 1.55 (m, 2H), 1.24 - 1.10 (m, 3H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESL') [(M+H)+]: 446.1.
Example 40 6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide 0 rr \
N
N.--N
*
N
H
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36g) and N-butyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide (32 mg, Example 40) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.28 - 7.14 (m, 4H), 6.88 (s, 2H), 4.95 (s, 2H), 4.16 (s, 1H), 3.41 - 3.36 (m, 2H), 3.10 - 2.99 (m, 3H), 2.53 - 2.51 (m, 2H), 2.27 (s, 3H), 1.71 - 1.63 (m, 2H), 1.62 - 1.51 (m, 2H), 1.42 - 1.26 (m, 2H), 0.97 - 0.74 (m, 6H). MS obsd. (ESI ) [(M+H)+]: 474.3 Example 41-A and Example 41-B
6-Amino-9-[(4-chlorophenyl)methy1]-2-IS(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A) and 6-Amino-9-[(4-chlorophenyl)methy1]-2-IS(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B) ,.., NH2'-'N N H2 N
NNo NI---"'N
OµN oL j., H 1\lµN s, I >==o S ' N N ,S
I-INV 0' Step 1: Preparation of 6-amino-9-[(4-chlorophenyl)methyl]-2-ethylsulfany1-7H-purin-8-one (Compound 41a) H
NCN
I
S)N N
41a Compound 41a was prepared in analogy to Example 1, Method A, Step 3 by using iodoethane and 6-amino-9-[(4-chlorophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 34b) instead of bromopropane and 6-amino-9-phenylmethy1-2-sulfany1-purin-8-one (Compound lb). 6-Amino-9-[(4-chlorophenyl)methy1]-2-ethylsulfany1-purin-8-one (2.5 g, Compound 41a) was obtained as a white solid. MS obsd.
(ESL') [(M+H)+]: 336.
Step 2: Preparation of 6-amino-9-(4-chlorobenzy1)-2-ethylsulfmy1-7H-purin-8-one (Compound 41b) H
I /C) ii it CI
41b Compound 41b was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-ethylsulfany1-7H-purin-8-one (Compound 41a) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-9-(4-chlorobenzy1)-2-ethylsulfiny1-7H-purin-8-one (1.94 g, Compound 41b) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 352.
Step 3: Preparation of 6-amino-9-[(4-chlorophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 41c) )Z
NN
SNN
NH
ilt CI
41c Compound 41c was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-9-(4-chlorobenzy1)-2-ethylsulfiny1-7H-purin-8-one (Compound 41b) instead of 6-amino-9-benzy1-2-(2-methylsulfiny1)-7H-purin-8-one (Compound 1d). 6-Amino-9-[(4-chlorophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (217 mg, Example 41c) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 10.61 (s, 1H), 7.42 -7.35 (m, 4H), 6.98 (s, 2H), 4.96 (s, 2H), 4.05 (s, 1H), 3.42 - 3.37 (m, 2H), 1.16 (t, J = 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 367Ø
Separation of compound of Compound 41c by chiral HPLC afforded Compound 41c-A (faster eluting, 31.8 mg) and Compound 41c-B (slower eluting, 10 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak IC-3 column.
N
FINN, ,S = N N
0' a 41c-A
Compound 41c-A: 1H NMR (400 MHz, DMSO-d6) gppm: 10.76 (s, 1H), 7.45 - 7.33 (m, 4H), 7.01 (s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.40 - 3.34 (m, 2H), 1.17 (t, J
= 7.4 Hz, 3H).
MS obsd. (ESL') [(M+H)+]: 367Ø
NN
)=0 ,S = N N
H
CI
41c-B
Compound 41c-B: 1H NMR (400 MHz, DMSO-d6) gppm: 10.70 (s, 1H), 7.46 -7.28 (m, 4H), 7.01 (s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.44 - 3.36 (m, 2H), 1.17 (t, J= 7.4 Hz, 3H).
MS obsd. (ESL') [(M+H)+]: 367Ø
Step 4: 6-Amino-9-[(4-chlorophenyl)methy1]-2-1S(R)-ethylsulfonimidoyll-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A) and 6-amino-9-[(4-chlorophenyl)methy1]-2-1S(S)-ethylsulfonimidoyll-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B) _ NI-12L*N N I-120 N
N N
N¨' N--' % ,I j., )=0 I-1 NN, ,, I
,s' s N N ,s' N N
I-11\V 0' * CI = CI
Example 41-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-9-[(4-chlorophenyOmethy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A, 78 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 5.00 (s, 2H), 4.19 (s, 1H), 3.46-3.39 (m, 2H), 3.39 - 3.38 (m, 2H), 3.09-2.99 (m, 3H), 1.69 -1.52 (m, 2H), 1.19 (t, J= 7.28 Hz, 3H), 0.95 - 0.66 (m, 3H). MS obsd. (ESL') [(M+H)+]:
466.1.
Example 41-B (125 mg) was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-purin-8-one (Compound 1 e ). 6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B, 38 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 5.00 (s, 2H), 4.20 (s, 1H), 3.46 - 3.41 (m, 2H), 3.40 -3.39 (m, 2H), 3.10 - 3.00 (m, 3H), 1.69 - 1.50 (m, 2H), 1.24 - 1.12 (m, 3H), 0.93 - 0.73 (m, 3H).
(MS obsd.
(ESL') [(M+H)+]: 466.2.
The stereochemistry of Example 41-B was determined by single crystal X-ray diffraction shown in Figure 1.
Example 42-A and Example 42-B
6-Amino-9-[(4-chlorophenyl)methyll-N-ethy1-2[S(S)-ethylsulfonimidoyll-N-methy1-oxo-purine-7-carboxamide (Example 42-A) and 6-Amino-9-[(4-chlorophenyl)methyll-N-ethy1-2-1S(R)-ethylsulfonimidoyll-N-methy1-8-oxo-purine-7-carboxamide (Example 42-B) , r N H2Ly N
r2uyN
N'N
N
I-1 NINN ,, I )= N -----,s . N N
,_'s -N ¨NN 0' H NV )0 CI
. ci Example 42-A was prepared in analogy to Example 1, Method A, step 6 by using Compound 41c-A and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide (Example 42-A, 40 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 4.99 (s, 2H), 4.18 (s, 1H), 3.48 - 3.40 (m, 2H), 3.39 (s, 2H), 3.05 ¨3.01 (m, 3H), 1.20 - 1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 452.2.
Example 42-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-B and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[5(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide (Example 42-B, 38 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.91 (s, 2H), 4.98 (s, 2H), 4.19 (s, 1H), 3.48 - 3.40 (m, 2H), 3.39 (s, 2H), 3.09 -2.97 (m, 3H), 1.23- 1.11 (m, 6H). MS obsd. (ESE') [(M+H)+]: 452.2.
The stereochemistry of Example 42-A was determined by single crystal X-ray diffraction shown in Figure 2.
Example 43-A and Example 43-B
6-Amino-2-1S(R)-ethylsulfonimidoy11-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A) and 6-Amino-2-1S(S)-ethylsulfonimidoy11-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B) NJ---N
oNN s, 0 NJ'N
N N H N
NN s, 0 S ' * NV ,S . N N
HI
0' Step 1: Preparation of 4-amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (Compound 43a) H
N
N _ 3__Nru N
*H 2N
43a Compound 43a was prepared in analogy to Example 1, Method A, Step 1 by using 4-methylbenzyl isocyanate instead of benzyl isocyanate. 4-Amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (26.6 g, Compound 43a) was obtained as a grey solid and used directly for next step without further purification. MS obsd.
(ESL') [(M+H)+]: 229.2.
Step 2: Preparation of 6-amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (Compound 43b) H
N )j1:1 NI\_o H S N N
*
43b Compound 43b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (compound 43a) instead of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (20.0 g, Compound 43b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 288.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43c) H
N
.SIN N
#
43c Compound 43c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (Compound 43b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (13 g, Compound 43c) was obtained as a yellow solid. MS obsd. (ESE) [(M+H)+]: 316.
Step 4: Preparation of 6-amino-2-ethylsulfmy1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43d) NI-1\11 SLI
N N
II
*
43d Compound 43d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43c) instead of 6-amino-9-benzy1-2-methylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-2-ethylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one6 (3.5 g, Compound 43d) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 332.
Step 5: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43e) H
NN
SN N
õ
N H
#
43e Compound 43e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-2-ethylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43d) instead of 6-amino-9-benzy1-2-methylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(ethylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (530 mg, Compound 43e) was obtained as a yellow solid. 1H NMR (400 MHz, DMSO-d6) gppm: 10.53 (s, 1H), 7.24 (d, J
= 8.03 Hz, 2H), 7.13 (d, J= 8.03 Hz, 2H), 6.94 (br. s., 2H), 4.91 (s, 2H), 4.03 (s, 1H), 3.36 -3.41 (m, 2H), 2.26 (s, 3H), 1.18 (t, J= 7.28 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 347.
Separation of compound of Compound 43e by chiral HPLC afforded Compound 43e-A (faster eluting, 56.8 mg) and Compound 43e-B (slower eluting, 56.7 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
H
N) --N
oNN 0 ,S
I-INV
=
43e-A
Compound 43e-A: 1H NMR (400 MHz, DMSO-d6) gppm: 10.52 (br. s., 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J = 7.9 Hz, 2H), 6.94 (br. s., 2H), 4.90 (s, 2H), 4.03 (s, 1H), 3.42 -3.33 (m, 2H), 2.25 (s, 3H), 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
347.
H
NN
FINN\ L I O
,S
0' y 43e-B
Compound 43e-B: 1H NMR (400 MHz, DMSO-d6) gppm: 10.56 (br. s., 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J = 8.0 Hz, 2H), 6.95 (br. s., 2H), 4.90 (s, 2H) 4.03 (s, 1H), 3.44 -3.29 (m, 2H), 2.25 (s, 3H), 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
347.
Step 6: Preparation of 6-Amino-2-1S(R)-ethylsulfonimidoyli-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A) and 6-Amino-2-1S(S)-ethylsulfonimidoyli-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B) _ d NI H2L*N
NH2uN
N
N I-1 N N..
).-0 s, 0 N
\NNN I 0 ,S ' N N
HIN
..s ' N N ,, 0' S .
Example 43-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A, 58.1 mg, faster eluting, isopropanol from 5% to 40% (0.05%DEA)/CO2 on ChiralPak AD-3 column) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.8 Hz, 2H), 7.15 (d, J=
7.8 Hz, 2H), 6.88 (br. s., 2H), 5.03 - 4.87 (m, 2H), 4.19 (s, 1H), 3.61 -3.36 (m, 4H), 3.11 ¨ 2.96 (m, 3H), 2.26 (s, 3H), 1.72 - 1.45 (m, 2H), 1.20 (t, J= 7.2 Hz, 3H), 0.97 - 0.65 (m, 3H). MS
obsd. (ESL') [(M+H)+]: 446.
Example 43-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B, 40.1 mg, slower eluting, isopropanol from 5% to 40% (0.05%DEA)/CO2 on ChiralPak AD-3 column) was obtained as a white solid: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.5 Hz, 2H), 7.15 (d, J =
7.5 Hz, 2H), 6.89 (br. s., 2H), 5.03 - 4.86 (m, 2H), 4.19 (s, 1H), 3.49 - 3.37 (m, 4H), 3.08 - 3.00 (m, 3H), 2.27 (s, 3H), 1.70 - 1.48 (m, 2H), 1.20 (t, J= 7.2 Hz, 3H), 0.95 - 0.71 (m, 3H). MS
obsd. (ESE') [(M+H)+]: 446.3.
The stereochemistry of Example 43-B was determined by single crystal X-ray diffraction shown in Figure 3.
Example 44-A and Example 44-B
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoyli-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-A) and 6-Amino-N-ethy1-2-1S(R)-ethylsulfonimidoyli-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-B) r NNµ oL
,S N N SN N
Example 44-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-B and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2[5(5)-ethylsulfonimidoy1]-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-A, 73.1 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.8 Hz, 2H), 7.15 (d, J= 7.8 Hz, 2H), 6.90 (br. s., 2H), 4.95 (s, 2H), 4.19 (br. s., 1H), 3.48 - 3.39 (m, 4H), 3.06 - 3.00 (m, 3H), 2.27 (s, 3H), 1.29 - 1.04 (m, 6H). MS obsd. (ESE') [(M+H)+]: 432.
Example 44-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-A and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzyl-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-B, 46.7 mg) was obtained as a white solid: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J= 7.9 Hz, 2H), 7.15 (d, J= 7.9 Hz, 2H), 6.90 (br. s., 2H), 4.95 (s, 2H), 4.19 (br. s., 1H), 3.50 - 3.39 (m, 4H), 3.10 ¨ 2.96 (m, 3H), 2.27 (s, 3H), 1.27- 1.10 (m, 6H). MS obsd. (ESL') [(M+H)+]:
432.
Example 45-A and Example 45-B
6-Amino-2-IS (R) ethylsulfonimidoy1]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide and 6-Amino-2-1S(S)ethylsulfonimidoy1]-9-1(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide HNµN
N N ,S N N
H
F F
Step 1: Preparation of 4-amino-3-1(4-fluorophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 45a) H2N * F
45a Compound 45a was prepared in analogy to Example 1, Method A, Step 1 by using 4-fluorobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-fluorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (48 g, Compound 45a) was obtained as a light yellow solid and was used directly for next step without further purification. MS obsd. (ESL') [(M+H)+]: 233.
Step 2: Preparation of 6-amino-9-[(4-fluorophenyl)methyl]-2-sulfany1-7H-purin-8-one (Compound 45b) H
NJ il I
H S C N N
* F
45b Compound 45b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-3-[(4-fluorophenyOmethy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 45a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-[(4-fluorophenyl)methy1]-2-sulfany1-7H-purin-8-one (32.0 g, Compound 45b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 292.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45c) N)j:N
SIN N
45c Compound 45c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-fluorophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 45b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (5.6 g, Compound 45c) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]:
320.
Step 5: Preparation of 6-amino-2-ethylsulfmy1-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45d) NJ11-1\11 S(I
N N
F
45d Compound 45d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-2-ethylsulfany1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45c) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-2-ethylsulfiny1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (4.8 g, Compound 45d) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 332.
Step 6: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45e) N
I
H N
* F
45e Compound 45e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-2-ethylsulfiny1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45d) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (2.9 g, Compound 45e) was obtained as a yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.57 (br.
s., 1H), 7.40 (dd, J= 8.5, 5.5 Hz, 2H), 7.16 (t, J= 8.9 Hz, 2H), 6.97 (br. s., 2H), 4.94 (s, 2H), 4.07 (s, 1H), 3.43 - 3.36 (m, 2H), 1.17 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 351.
Separation of compound of Compound 45e by chiral HPLC afforded Compound 45e-A (faster eluting, 85.4 mg) and Compound 45e-B (slower eluting, 36.4 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 45e-A: 1H NMR (400 MHz, DMSO-d6)6 ppm: 10.53 (br. s., 1H), 7.41 (dd, J=
8.5, 5.5 Hz, 2H), 7.17 (t, J= 8.9 Hz, 2H), 6.98 (br. s., 2H), 4.95 (s, 2H), 4.07 (s, 1H), 3.45-3.36 (m, 2H), 1.17 (t, J = 7.3 Hz, 3H). MS obsd. (ES[) [(M+H)+]: 351.
Compound 45e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.53 (br. s., 1H), 7.41 (dd, J=
8.5, 5.5 Hz, 2H), 7.17 (t, J= 8.9 Hz, 2H), 6.98 (br. s., 2H), 4.95 (s, 2H), 4.07 (s, 1H), 3.44 -3.37 (m, 2H) 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 351.
Step 7: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 45), 6-Amino-2-IS (R)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-Amino-2-1S(S)ethylsulfonimidoyl]-9-1(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 45-A and Example 45-B).
N I-12L* N
ON, I
N
I-IN' F
(Example 45) NI11-12`-*N
0N\ I I 1-INNµ I )=0 N N SN N
HN- 0' y F = F
(Example 45-A and Example 45-B) Example 45 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45e) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (162.4 mg, Example 45) was obtained as a white solid.
Separation of compound of Example 45 by chiral HPLC afforded Example 45-A
(faster eluting, 85.3 mg) and Example 45-B (slower eluting, 52 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 45-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.53 -7.38 (m, 2H), 7.18 (t, J= 8.9 Hz, 2H), 6.90 (br. s., 2H), 4.99 (s, 2H), 4.21 (s, 1H), 3.48 - 3.37 (m, 4H), 3.10 -3.01 (m, 3H), 1.69 - 1.49 (m, 2H), 1.25 - 1.14 (m, 3H), 0.94 - 0.72 (m, 3H).
MS obsd.
(ESL') [(M+H)+]: 450.
Example 45-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.54 - 7.38 (m, 2H), 7.18 (t, J= 8.9 Hz, 2H), 7.01 - 6.72 (m, 2H), 4.99 (s, 2H), 4.21 (s, 1H), 3.46 - 3.38 (m, 4H), 3.10 -3.01 (m, 3H), 1.76 - 1.50 (m, 2H), 1.25 - 1.16 (m, 3H), 0.99 - 0.69 (m, 3H).
MS obsd.
(ESI ) [(M+H)+]: 450.
Example 46-A and Example 46-B
6-Amino-N-ethyl-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methyl-8-oxo-purine-7-carboxamide (Example 46), 6-amino-N-ethyl-2-1S(S)-(ethylsulfonimidoy1)1-9-[(4-fluorophenyl)methy1]-N-methyl-8-oxo-purine-7-carboxamide and 6-amino-N-ethyl-2-1S(R)-(ethylsulfonimidoy1)1-9-[(4-fluorophenyl)methyl]-N-methyl-8-oxo-purine-7-carboxamide (Example 46-A and Example 46-B).
r NH20yN
N
N"
oNµ I 0 SN N
. F
(Example 46) r r NI-12L', N NH2 N
N.,....N N N
J----H NNµ L 1 oNN s,I I 0 ,S ' N N S ' N N
0' y H NV
. F / . F
(Example 46-A and Example 46-B) Example 46 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45e) and N-ethyl-N-methyl carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-purine-7-carboxamide (51 mg, Example 46) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.15 (m, 2H), 6.90 (br. s., 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.47 - 3.32 (m, 4H), 3.05 -3.01 (m, 3H), 1.21 - 1.14 (m, 6H). MS obsd. (ESI ) [(M+H)+]: 436.
Separation of compound of Example 46 by chiral HPLC afforded Example 46-A
(faster eluting, 72 mg) and Example 46-B (slower eluting, 45 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 46-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.16 (m, 2H), 6.90 (br. s., 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.47 - 3.32 (m, 4H), 3.05 - 3.01 (m, 3H), 1.21-1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 436.
Example 46-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.14 (m, 2H), 6.92 (br. s., 2H), 4.98 (s, 2H), 4.20 (br. s., 1H), 3.47 - 3.32 (m, 4H), 3.05 -3.01 (m, 3H), 1.23 - 1.19 (m, 6H). MS obsd. (ESL') [(M+H)+]: 436.
Example 47-A and Example 47-B
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47), 6-amino-2-IS(R)-ethylsulfonimidoy1]-[(4-bromophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-amino-2-IS(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide NFIPN
N------N
,sN N
HN' . Br (Example 47) N1-12 N NHPyN
N--N N)--' N
% s,I )=0 1-1N (:) S
,,L I
S ' N N s' NN
HNV CV
0 Br . Br (Example 47-A and Example 47-B) Step 1: Preparation of 4-amino-3-[(4-bromophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 47a) N H
. Br 47a Compound 47a was prepared in analogy to Example 1, Method A, Step 1 by using 4-bromobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-bromophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (500 mg, Compound 47a) was obtained as a light yellow solid and was used directly for next step without further purification. 1H NMR (400 MHz, DMSO-d6) 5 ppm: 9.94 (S, 1H), 7.55-7.53 (d, J=
8.0 Hz, 2H), 7.20-7.18 (d, J = 8.0 Hz, 2H), 6.52 (br. s., 2H), 4.74 (s, 2H). MS obsd.
(ESI ) [(M+H)+]: 293.
Step 2: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 47b) N- N
1 )=0 HS NN
. Br 47b Compound 47b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-3-[(4-bromophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 47a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (300 mg, Compound 47b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 352.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-[(4-bromophenyl)methyl]-7H-purin-8-one (Compound 47c) Ni Nso S NN
) *Br 47c Compound 47c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 45b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-[(4-bromophenyOmethyl]-7H-purin-8-one (5.6 g, Compound 47c) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]:
380.
Step 4: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfmyl-7h-purin-8-one (compound 47d) N:N
S)N N
ii * Br 47d Compound 47d was prepared in analogy to Example 1, Method B, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfany1-7H-purin-8-one ( Compound 47c) instead of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound 1c).
Amino-9-[(4-bromophenyl)methy1]-2-ethylsulfiny1-7H-purin-8-one (3.2 g, Compound 47d) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 396.
Step 5: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7h-purin-8-one (compound 47e) H
N N
0 1 >0 \ \ ;.........
H N= S N N
) . Br 47e Compound 47e was prepared in analogy to Example 1, Method B, Step 7 by using 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfiny1-7H-purin-8-one (Compound 47d) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (4.0 g, Compound 47e) was obtained as a white solid. MS obsd. (ESE') [(M+H)+]: 411.
N NH N
ssL 1-1No I
S N S N N
H N 0' = Br 1>
Br Compound 47e-A and Compound 47e-B
Separation of compound of Compound 47e by chiral HPLC afforded Compound 47e-A (faster eluting, 112 mg) and Compound 47e-B (slower eluting, 99 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 47e-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.58 (br. s., 1H), 7.52-7.54 (d, J= 8.0, 2H), 7.31-7.29 (t, J= 8.0 Hz, 2H), 6.54 (br. s., 2H), 4.93 (s, 2H), 4.05 (s, 1H), 3.42 -3.31 (m, 2H), 1.15 (t, J= 7.3 Hz, 3H). MS obsd. (ESI ) [(M+H)+]: 411.
Compound 47e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.58 (br. s., 1H), 7.54-7.52 (d, J= 8.0, 2H), 7.31-7.29 (t, J= 8.0 Hz, 2H), 6.98 (br. s., 2H), 4.93 (s, 2H), 4.06 (s, 1H), 3.40 -3.37 (m, 2H), 1.15 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 411.
Step 6: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47), 6-amino-9-[(4-bromophenyl)methy1]-2-1S(R)-ethylsulfonimidoyli-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-amino-9-[(4-bromophenyl)methy1]-2-1S(S)-ethylsulfonimidoyli-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47-A and Example 47-B).
N I-12L*
NN
I-11\V
= Br (Example 47) NH2uy N"
oL HNx, N N
Br Br (Example 47-A and Example 47-B) Example 47 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 47e) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (570 mg, Example 47) was obtained as a white solid. 1H
NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.53 (m, 2H), 7.36-7.34 (m, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.18 (s, 1H), 3.45 - 3.38 (m, 4H), 3.05 - 3.02 (m, 3H), 1.65-1.56 (m, 2H), 1.19 (t, J= 8.0 Hz, 3H), 0.93-0.75 (m, 3H). MS obsd. (ESL') [(M+H)+]: 510.
Separation of compound of Example 47 by chiral HPLC afforded Example 47-A
(faster eluting, 260 mg) and Example 47-B (slower eluting, 266 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 47-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.33 (d, J= 8,0 Hz, 2H), 6.90 (br. s., 2H), 4.97 (s, 2H), 4.21 (s, 1H), 3.46 - 3.41 (m, 4H), 3.05 - 3.02 (m, 3H),1.65-1.54 (m, 2H), 1.24-1.16 (m, 3H), 0.93-0.75 (m, 3H). MS
obsd. (ESE') [(M+H)+]: 510.
Example 47-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.54 - 7.53 (d, J= 8.0 Hz, 2H), 7.36-7.33 (d, J= 8,0 Hz, 2H), 6.90 (br. s., 2H), 4.97 (s, 2H), 4.21 (s, 1H), 3.46 - 3.41 (m, 4H), 3.06 - 3.02 (m, 3H), 1.65-1.54 (m, 2H), 1.20-1.16 (m, 3H), 0.93-0.75 (m, 3H).
MS obsd. (ESI ) [(M+H)+]: 510.
Example 48-A and Example 48-B
6-Amino-9-[(4-bromophenyl)methy1]-N-ethyl-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide (Example 48), 6-amino-9-[(4-bromophenyl)methy1]-N-ethyl-2-IS(S)-(ethylsulfonimidoy1)1-N-methyl-8-oxo-purine-7-carboxamide and 6-amino-9-[(4-bromophenyl)methyll-N-ethy1-2-1S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide (Example 48-A and Example 48-B).
NH2 o.....r T \
N-1\1 SN----N
II
= Br (Example 48) 0 r---NH2 .......N N H2 o.....r \ \
NJ-----N N----"N
N H4 (:) 0 1 >=0 0,1, , H Nk I I
S 's N-----N
= Br ic = Br (Example 48-A and Example 48-B) r r NH2 yN
NH2 ,,N
N)-----N 111\1µµ .0L I )=C) NN
-""k 0 0' S N N
H NV
ilt CI
Example 48 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 47e) and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-bromophenyOmethyl]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (469 mg, Example 48) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 -7.54 (d, J
= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.98 (br. s., 2H), 4.97 (s, 2H), 3.53 - 3.46 (m, 4H), 3.05 - 3.01 (m, 3H), 1.22-1.16 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Separation of compound of Example 48 by chiral HPLC afforded Example 48-A
(faster eluting, 198 mg) and Example 48-B (slower eluting, 202 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Example 48-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.19 -4.18 (m, 1H), 3.46 - 3.41 (m, 4H), 3.05 - 3.01 (m, 3H), 1.20-1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Example 48-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.24 (br.
s., 1H), 3.58 -3.41 (m, 4H), 3.05 - 3.01 (m, 3H), 1.26-1.01 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Example 49 Activity of Compounds and Examples in HEK293-hTLR-7 assay HEK293-Blue-hTLR-7 cells assay:
A stable HEK293-Blue-hTLR-7 cell line was purchased from InvivoGen (Cat.#:
hkb-ht1r7, San Diego, California, USA). These cells were designed for studying the stimulation of human TLR7 by monitoring the activation of NF-KB. A SEAP
(secreted embryonic alkaline phosphatase) reporter gene was placed under the control of the IFN-13 minimal promoter fused to five NF-KB and AP-1-binding sites. The SEAP was induced by activating NF-KB and AP-1 via stimulating HEK-Blue hTLR7 cells with TLR7 ligands.
Therefore the reporter expression was regulated by the NF-KB promoter upon stimulation of human TLR7 for 20 hrs. The cell culture supernatant SEAP reporter activity was determined using QUANTI-BlueTm kit (Cat.#: rep-qbl, Invivogen, San Diego, Ca, USA) at a wavelength of 640 rim, a detection medium that turns purple or blue in the presence of alkaline phosphatase.
HEK293-Blue-hTLR7 cells were incubated at a density of 250,000-450,000 cells/mL in a volume of 180 [EL in a 96-well plate in Dulbecco's Modified Eagle's medium (DMEM) containing 4.5 g/L glucose, 50 U/mL penicillin, 50 mg/mL streptomycin, mg/mL Normocin, 2 mM L-glutamine, 10% (V/V) heat-inactivated fetal bovine serum for 24 hrs. Then the HEK293-Blue-hTLR-7 cells were incubated with addition of 20 [EL test compound in a serial dilution in the presence of final DMSO at 1% and perform incubation under 37 C in a CO2 incubator for 20 hrs. Then 20 [EL of the supernatant from each well was incubated with 180 [EL Quanti-blue substrate solution at 37 C for 2 hrs and the absorbance was read at 620-655 nm using a spectrophotometer. The signalling pathway that TLR7 activation leads to downstream NF-KB activation has been widely accepted, and therefore similar reporter assay was also widely used for evaluating TLR7 agonist (Tsuneyasu Kaisho and Takashi Tanaka, Trends in Immunology, Volume 29, Issue 7, July 2008, Pages 329.sci; Hiroaki Hemmi et al, Nature Immunology 3, 196 - 200 (2002)).
The Compounds and Examples of the present invention were tested in HEK293-hTLR-7 assay for their TLR7 agonism activity as described herein and results are listed in Table 1. The Examples of prodrugs were found to have EC50 of about 2.1 [EM to about 1000 [EM, the Compounds of active forms were found to have EC50 less than 0.2 [M. The calculated ratio of ECso(prodnio / ECso(active form) were within the range from 32 to about 7600.
Table 1. Activity of Examples and Compounds of present invention in HEK293-hTLR-7 assay Ratio hTLR-7 EC50 hTLR-7 EC50 Corresponding Prodrug (EC50(prodrug) /
Active Form (Prodrug, (Active form, EC50(active form)) ILLM) tiM) Example 1 50.4 Compound le 0.065 775.4 Example 1-A 42.5 Compound le-A 0.067 634.3 Example 1-B 27 Compound le-B 0.086 314.0 Example 2 32 Compound le 0.065 372.1 Example 2-A 3.7 Compound le-B 0.086 43.0 Example 2-B 4.4 Compound 1 e-A 0.067 65.7 Example 3 15.1 Compound le 0.065 232.3 Example 4 23 Compound le 0.065 353.8 Example 5 41 Compound le 0.065 630.8 Example 6 82.3 Compound le 0.065 1266.2 Example 7 19.9 Compound le 0.065 306.2 Example 8 2.1 Compound le 0.065 32.3 Example 9 19.2 Compound le 0.065 295.4 Example 10 68.5 Compound le 0.065 1053.8 Example 11 5.6 Compound le 0.065 86.2 Example 12 43.9 Compound le 0.065 675.4 Example 13 67 Compound le 0.065 1030.8 Example 14 2.4 Compound le 0.065 36.9 Example 15 494 Compound le 0.065 7600 Example 16 32.1 Compound le 0.065 493.8 Example 25 24.2 Compound le 0.065 372.3 Example 26 13.4 Compound le 0.065 206.2 Example 27 31.7 Compound le 0.065 487.7 Example 28 6.9 Compound le 0.065 106.2 Example 29 48.8 Compound le 0.065 750.8 Example 32 22.5 Compound le 0.065 346.2 Example 34- 6.0 Compound 34e-A 0.014 428.6 A
Example 34-6.36 Compound 34e-B 0.011 578.2 B
Example 36-A 31.8 Compound 36g-A 0.019 1673.7 Example 37-A 26.6 Compound 36g-A 0.019 1400 Example 37-B 47.4 Compound 36g-B 0.022 2154.5 Example 38-A 26.2 Compound 36g-A 0.019 1378.9 Example 38-B 19.5 Compound 36g-B 0.022 886.4 Example 39 4.3 Compound 36g 0.027 159.3 Example 40 52.8 Compound 36g 0.027 1955.6 Example 41 36 Compound 41c 0.053 679.2 Example 41-44.1 Compound 41c-B 0.085 518.8 A
Example 41-32.1 Compound 41c-A 0.071 452.1 B
Example 42-40.5 Compound 41c-A 0.071 570.4 A
Example 42-49.2 Compound 41c-B 0.085 578.8 B
Example 43-110 Compound 43e-A 0.11 1000 A
Example 43- 78.4 Compound 43e-B 0.035 2240 B
Example 44-65.4 Compound 43e-B 0.035 1868.6 A
Example 44-96.7 Compound 43e-A 0.11 879.1 B
Compound 45e-B
Example 45-153 Or 0.26 or 0.39 588 or 392 A
Compound 45e-A
Compound 45e-B
Example 45->1000 Or 0.26 or 0.39 >3846 or >2564 B
Compound 45e-A
Compound 45e-A
Example 46-45.5 Or 0.26 or 0.39 175 or 116.7 A
Compound 45e-B
Compound 45e-B
Example 46-45.7 Or 0.26 or 0.39 175.7 or 117.2 B
Compound 45e-A
Compound 47e-A
Example 47-10.9 Or 0.021 or 0.025 519.0 or A
Compound 47e-B
Compound 47e-A
Or Example 47-13.1 0.021 or 0.025 623.8 or B Compound 47e-B
Compound 47e-A
Example 48-18.3 Or 0.021 or 0.025 871.4 or A
Compound 47e-B
Compound 47e-A
Or Example 48-20.8 0.021 or 0.025 990.5 or B Compound 47e-B
Example 50 Metabolism of prodrugs of compound of formula (I) A study was undertaken to evaluate the metabolic conversion of prodrugs, compound of formula (I), to its corresponding active form. The compounds of formula (I), if served as prodrugs, can be metabolized to the active compound or other compounds of the invention in the body. Human liver microsomes are often used to assess the degree of metabolic conversion of prodrugs in the body of animal or human.
Materials NADPH cofactor system including 13-Nicotinamide adenine dinucleotide phosphate (NADP), isocitric acid and isocitric dehydrogenase were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Human liver microsomes (Cat No. 452117, Lot No.
38290) were obtained from Corning (Woburn, MA, USA). Mouse liver microsomes (Cat No.
M1000, Lot No.1310028) were obtained from Xenotech.
Working solution of the compounds and other solution Compounds were dissolved in DMSO to make 10 mM stock solutions. 10 [EL of the stock solution was diluted with acetonitrile (990 [EL) to get a 100 [EM
working solution.
Incubation Microsomes were preincubated with test compound for 10 min at 37 C in 100 mM
potassium phosphate buffer with pH 7.4. The reactions were initiated by adding NADPH
regenerating system to give a final incubation volume of 200 [EL and shaken in a water bath at 37 C. Incubation mixtures consisted of liver microsomes (0.5 mg microsomal protein/mL), substrates (1.0 [tM), and NADP (1 mM), isocitric dehydrogenase(1 unit/mL), isocitric acid (6 mM).
Preparation of Samples for Analysis At 30 min, reaction was quenched by adding 600 [LL cold acetonitrile (including 100 ng/mL tolbutamide and 100 ng/mL labetalol as internal standard). The samples were centrifuged at 4000 rpm for 20 minutes and the resultant supernatants were subjected to LC-MS/MS analysis.
The samples for calibration curve were prepared as followed. Dispense 100 [LL/well liver microsomes and 98 [LL/well NADPH regenerating system solution to 96-well plate.
Add 600 [LL quenching solution first, and then followed by 2 [EL Standard curve and QC
working solution.
Bioanalysis The compounds were quantified on an API4000 LC-MC/MC instrument in the ESI-Positive MRM mode.
A study was undertaken to evaluate the metabolic conversion of prodrugs (1[EM), Example 1, Example 1-A, Example 1-B, Example 2, Example 2-A, Example 2-B, Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, Example 9, Example 10, Example 11, Example 12, Example 13, Example 14, Example 15, Example 16, Example 17, Example 21, Example 22, Example 23, Example 25, Example 26, Example 27, Example 28Example 29, Example 30, Example 31, Example 32, Example 33, Example 34-A, Example 34-B, Example 36-A, Example 36-B, Example 37-A, Example 37-B, Example 38-A, Example 38-B, Example 39, Example 40, Example 41, Example 41-A, Example 41-B, Example 42, Example 42-A, Example 42-B, Example 43, Example 43-A, Example 43-B, Example 44, Example 44-A, Example 44-B and Example 45-A, Example 46-A, Example 46-B, Example 47-A, Example 47-B, Example 48-A, Example 48-B to the corresponding active forms, Compound le, Compound le-A, Compound le-B, Compound 34e-A, Compound 34e-B, Compound 36g-A, Compound 36g-B, Compound 36g, Compound 41c, Compound 41c-B, Compound 41c-A, Compound 43e, Compound 43e-A, Compound 43e-B, Compound 45e-A, Compound 45e-B, Compound 47e-A, and Compound 47e-B in the presence of human liver microsomes. Results were summarized and shown in Table 2.
Table 2. Metabolic conversion of prodrugs in human liver microsomes Metaboli Metaboli zed zed product product Corresponding Corresponding concentr concentr Example Metabolized Metabolized ation in Example No.
ation in No. Product (active Product (active human human form) form) liver liver microso microso mes (?LM) mes (?LM) Example 1 Compound le 0.0214 Example 31 Compound le 0.005 Example 1- Compound le-A 0.018 Example 32 Compound le 0.013 Example 1- Compound le-B 0.022 Example 33 Compound le 0.59 Example 2 Compound le 0.028 Example 34-A Compound 34e-A 0.2 Example 2- Compound le-B 0.036 Example 34-B Compound 34e-B 0.088 Example 2- Compound le-A 0.029 Example 36-A Compound 36g-A 0.02 Example 3 Compound le 0.12 Example 36-B Compound 36g-B 0.019 Example 5 Compound le 0.078 Example 37-A Compound 36g-A 0.004 Example 6 Compound le 0.074 Example 37-B Compound 36g-B 0.002 Example 7 Compound le 0.15 Example 38-A Compound 36g-A 0.026 Example 8 Compound le 0.043 Example 38-B Compound 36g-B 0.034 Example 9 Compound le 0.002 Example 40 Compound 36g 0.032 Example 10 Compound le 0.005 Example 41-A Compound 41c-B 0.38 Example 11 Compound le 0.001 Example 41-B Compound 41c-A 0.36 Example 12 Compound le 0.018 Example 42-A Compound 41c-A 0.14 Example 13 Compound le 0.04 Example 42-B Compound 41c-B 0.004 Example 14 Compound le 0.026 Example 43-A Compound 43e-A 0.014 Example 15 Compound le 0.002 Example 43-B Compound 43e-B 0.016 Example 16 Compound le 0.024 Example 44-A Compound 43e-B 0.002 Example 17 Compound le 0.075 Example 44-B Compound 43e-A 0.002 Example 21 Compound le 0.48 Example 45-A Compound 45e-B 0.41 Or Example 22 Compound le 0.42 Example 46-A Compound 45e-A 0.039 Or Example 23 Compound le 0.42 Example 46-B Compound 45e-B 0.18 Or Example 25 Compound le 0.018 Example 47-A Compound 47e-A 0.36 Or Example 26 Compound le 0.042 Example 47-B Compound 47e-B
0.41 Or Example 27 Compound le 0.11 Example 48-A Compound 47e-A 0.11 Or Example 28 Compound le 0.084 Example 48-B Compound 47e-B
0.053 Or Example 29 Compound le 0.009 Example 51 In vivo combined efficacy (tumor free mice) of an prodrug form of compounds of the present invention (Compound 41-A) and sorafenib in a highly aggressive model of hepatocellular carcinoma In iAST mice tumorigenesis was initiated by intravenous injection of 5x108 IFU
adenovirus expressing Cre recombinase (Ad-CMV-iCre vector in vivo application, Vector Biolabs) into transgenic mice expressing the hepatocyte-specific albumin promoter, a loxP-flanked stop cassette, and the SV40 large T-antigen (Runge A, at al., Cancer Res. 74 (2014) 4157-69). The Cre recombinase excises the stop cassette in transduced cells and leads to a transient viral hepatitis and resulting in multinodular tumorigenesis within 8 weeks.
Female mice were treated with either vehicle (7.5% Gelatine / 0.22% NaCl for Sorafenib;
or 2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% Methylparaben (Sigma), 0.01%
Propylparabens (Sigma) in water for 41-A), or 90mg/kg in Sorafenib (Nexavar 0, Bayer HealthCare) daily or were treated with compound 41-A (10mg/kg) once a week by oral gavage. Treatment using vehicle or Sorafenib started on week 7.5 upon adenovirus administration and 3 days prior to administration compound 41-A. Animals were sacrificed on day 12 after treatment start and total liver and tumor weights were determined. Per group n=10 were analyzed by One-way ANOVA and Tukey correction shown as individual dots with means SEM using GraphPad Prism software version 6.
Although Sorafenib was highly effective in monotherapy, the combination with an active form of the compounds of the present invention (compound 41-A) resulted even in 2/10 tumor-free mice by superficial examination of the livers in this highly aggressive model of hepatocellular carcinoma. Results are shown in the Table below and in Figures lA and 1B
Synergistic effect of Compound 41-A and sorafenib on tumor burden ( tumor free mice) Treatment Examination of the liver for superficial tumor nodules Vehicle 0/10 tumor nodule free Compound 41-A 0/10 tumor nodule free Sorafenib 0/10 tumor nodule free Compound 41-A + 2/10 tumor nodule free Sorafenib Example 52 Treatment with an prodrug form of the compounds of the present invention (compound 41-A) induces PD-Li expression on tumor cells in hepatocellular carcinoma.
Tumors from iAST mice were treated as described in Figure 1. Animals were sacrificed on day 12 after treatment start and tumors analyzed by flow cytometry. For flow cytometry, tumors were excised and single cell suspensions obtained by mechanical processing and enzymatic digestion (DNAse 0.01%, Collagenase IV lmg/m1). Staining procedures started with Fc receptor blocking using 2.4G2 antibody clone (1:200 dilution, BD
Bioscience), and the following antibodies (clones) were used to analyzed leukocyte infiltrate: CD45-FITC (30-F11, BioLegend) and CD1 lb-BUV737 (M1/70, BD Bioscience). Samples were acquired using a LSR Fortessa machine (BD Bioscience) and analyzed by FlowJo version 10 (Treestar). Data are shown of n=5 per group, analyzed by One-way ANOVA and Tukey correction shown as individual dots with means SEM using GraphPad Prism software version 6. Although the absolute immune cell infiltrate in iAST tumor did not change by any treatments described (Fig.2A), significant changes were observed in the overall lymphoid and myeloid composition of the tumors (Fig. 2 C and D). Here, the changes were clearly driven by Sorafenib, which was previously shown to act also on immune cells (Martin del Campo, et al, J Immunol. 195 (2015) 1995-2005). 41-A treatment however induced PD-Li expression on tumor cells in monotherapy as well as in combination with Sorafenib (Fig. 2 B).
Example 53 In vivo triple combination of 41-A, Sorafenib and anti-PD-1 results in increased median survival.
Multinodular tumors were induced in iAST mice as described for Fig. 1 ( see Example 51).
Female transgenic mice were treated at 7.5 weeks upon virus injection with either vehicle (7.5% Gelatine / 0.22% NaCl for Sorafenib; or 2% Kluce10 Hydroxypropylcellulose LF
(Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% Methylparaben (Sigma), 0.01% Propylparabens (Sigma) in water for 41-A), or 90mg/kg in Sorafenib (Nexavar 0, Bayer HealthCare) daily or were treated with compound 41-A (10mg/kg) once a week by oral gavage. Treatment using vehicle or Sorafenib started on week 7.5 upon adenovirus administration and 3 days prior to administration compound 41-A. Anti-mouse PD-1 antibody (clone RPM1-14, BioXCell) was administered every 3 days intra peritoneally at 250[Eg/mouse. Total treatment period was 2 weeks (and 3 days + 2 weeks for Sorafenib) and survival of iAST mice was monitored. Mice were sacrificed upon display of distress signs such as >20%
gain of body weight, ruffled fur and or hatched position. Kaplan-Meier curves were analyzed by Pairwise Log-Rank test (see table). In the survival setting, neither Sorafenib, nor 41-A
were effective in monotherapy. Anti-PD-1 monotherapy even resulted in significantly reduced survival compared to VEH control. The median survival of iAST mice was significantly enhanced in the combination group of Sorafenib and anti-PD-1 antibody.
However, the triple combination of 41-A together with Sorafenib and anti-PD-1 led to the biggest and significant increase in median survival from 71 days (VEH) to 104 days (41-A
+ PD-1 + Sorafenib) in this highly aggressive HCC model. Results are shown in Figure 3 and the Table below Table: Pairwise Log-Rank Test (multiple test level = 0.00179) A +
+
Group VEH Sorafenib 41-A PD-1 41-A+ + PD- PD-1 PD-1 +
Sorafenib Sorafenib Sorafenib VEH 1.0 0.1580 0.2309 0.0004* 0.8759 0.1465 0.0192* 0.0001*
Example 54 Treatment with an prodrug form of the compounds of the present invention (compound 41-A) in the transplanted Hep55.1c mouse model of hepatocellular carcinoma Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20itl (10itl cell suspension plus 10itl Martigel).
Tumor volume was monitored weekly using itCT (TomoScope Synergy Twin, CT
Imaging GmbH) upon a single intravenous administration of contrasting agent Exitron (Viscovert). Imaging data were reconstructed by TomoScope software and analyzed using Osirix software. Once tumors reached 80mm3, mice were treated weekly with either 10mg/kg 41-A compound or vehicle (2% Kluce10 Hydroxypropylcellulose LF
(Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09%
Methylparaben (Sigma), 0.01% Propylparabens (Sigma) in water) per oral gavage.
To compare to another agonistic, immune stimulating agent, a single dose of anti-antibody (4mg/kg; clone FGK.45, BioXCell) was given. Data depicted are means SEM
for a minimum of n=9 animals per group.
Weekly administration of compound 41-A resulted in inhibition of tumor growth in Hep55.1c tumor bearing mice when compared to vehicle treatment. As previously published, a single dose of anti-CD40 antibody can lead to tumor eradication in subcutaneous MC38 tumors and it has been shown that the anti-CD40 antibody has an inflammatory effect in the liver (Hoves S, et al, J Exp Med, DOI:
10.1084/jem.20171440;
Published February 7, 2018). However, no beneficial treatment effect was observed in Hep55.1c tumor bearing mice with anti-CD40 antibody. Results are shown in Figure 5 A.
Example 55 In vivo efficacy of compound 42-A (6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide), alone and in combination with anti-PD-1 results in survival benefit in the Hep55.1c mouse model of hepatocellular carcinoma.
Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20 1 (10 1 cell suspension plus 10 1Martigel).
Scout animals were sacrificed to determine the time point of treatment start at about 80mm3 tumor volume. Mice were treated with either compound 42-A 10mg/kg or vehicle (2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% methylparaben (Sigma), 0.01%
propylparaben (Sigma) in water) per oral gavage, or intra peritoneal administration of 250[tg anti-PD-1 antibody (clone RPM1-14, BioXCell), or a combination of compound 42-A plus anti-PD-1. 42-A was given weekly (total 3 times) and stated on the same day anti-PD-1 antibody treatment. Antibody treatment was continued every three to four days for 6 doses in total. Monotherapy with 42-A resulted smaller tumor volume compared to VEH
control and PD-1 monotherapy. Combined treatment of 42-A and anti-PD-1, tumor volume was also reduced with 3 out of 9 mice being tumor-free. Results are shown in Figure 5 B
and the Table below.
Combination effect of Compound 42-A and anti-PD-1 on tumor burden (tumor free mice) Treatment Examination of the liver Vehicle 0/7 tumor free aPD-1 1/9 tumor free 42-A 1/7 tumor free 42-A+aPD-1 3/9 tumor free Example 56 Combination of an prodrug form of the compounds of the present invention (compound 41-A) and anti-PD-1 antibodies Hep55.1c mouse model of hepatocellular carcinoma.
Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20u1 (10u1 cell suspension plus 10 1 Martigel).
After 3 weeks, animals were sacrificed and tumors excised from the liver.
Excised tumors were cut into lx1 mm3 pieces and implanted into the liver of female C57BL/6N
mice.
Scout animals were sacrificed to determine the time point of treatment start at about 80mm3 tumor volume. Mice were treated with either 41-A or vehicle (2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol succinate (TPGS, Sigma), 0.09% methylparaben (Sigma), 0.01% propylparaben (Sigma) in water) per oral gavage, or intra peritoneal administration of 250ug anti-PD-1 antibody (clone RPM1-14, BioXCell), or a combination of 41-1 plus anti-PD-1. 41-A was given weekly, while anti-PD-1 antibody treatment started one day after 41-A
treatment and was continued every three to four days for 8 doses in total. Treatment with both agents was stopped after the last anti-PD-1 administration. Monotherapy with 41-A
resulted in longer survival of mice (5/10) compared to vehicle control (1/10). Combined treatment of 41-A
and anti-PD-1 enhanced survival of mice even significantly to 8/10 being alive on day 94 after tumor fragment transplantation.
Example 57 Treatment with an active form of the compounds of the present invention (compound 41c-B) does not induce enhanced tumor cell proliferation in cell lines originating from hepatocellular carcinoma and cholangiocarcinoma Cell lines derived from hepatocellular carcinoma and cholangiocarcinoma (EGI1 and OZ) were maintained and tested in the following media: Huh7 and EGI1 were cultured in DMEM 4,5 g/L glucose (Gibco, Cat# 31966-021), 10 % FCS (GIBCO, Cat# 10500-064 Lot 07G3690K), 2 mM L-glutamine (Thermo Fischer, Cat# 25030081), 1mM sodium pyruvate (GIBCO Cat# 11360-039). Hep3B and HepG2 were cultivated in Eagles MEM
+
Earle's BSS (PAN, Cat# PO4-08510), 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA
(PAN Cat# P08-32100) and 1 mM sodium pyruvate. JHH1, JHH5, JHH6 and OZ were cultivated in Williams'E (PAN Cat# PO4-29050), 10 % FCS and 2 mM L-glutamine.
was cultivated using Williams'E, 10 % FCS and 2 mM L-glutamine. HLE was cultivated in DMEM 4,5 g/L glucose, 10 % FCS and 2 mM L-glutamine. HLF was cultivated in DMEM
4,5 g/L glucose, 5 % FCS, 0.1 mM NEAA and 2 mM L-glutamine. JHH4 was cultivated in Eagles MEM + Earle's BSS, 10 % FCS and 2 mM L-glutamine. SkHepl was cultivated in Eagles MEM + Earle's BSS, 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA and 1mM
sodium pyruvate. SNU449 was cultivated using RPMI 1640 (PAN Cat# PO4-18047) 10 %
FCS and 2 mM L-glutamine. Cells were seeded in the respective media overnight at a density of 5,000 cells per well in 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, logarithmic dilutions of 4 1 c-B starting from 27[EM down to 270pM were added and incubated for 72, 120 and 148 hours, respectively. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 dye (2[Eg/ml, Sigma Cat# B2261). Data shown are means + SD
from triplicate wells based on the analysis of 9 images per well relative to the DMSO control.
None of the cell lines tested showed a significant increase in proliferation upon treatment with 41c-B directly at the depicted time points. Results are shown in Figure 6A.
Example 58 Treatment with an active form of the compounds of the present invention (compound 41c-A) does not induce enhanced tumor cell proliferation in cell lines originating from hepatocellular carcinoma and cholangiocarcinoma Cell lines derived from hepatocellular carcinoma and cholangiocarcinoma (EGI1) were maintained and tested in the following media: Huh7 and EGI1 were cultured in DMEM 4,5 g/L glucose (Gibco, Cat# 31966-021), 10 % FCS (GIBCO, Cat# 10500-064 Lot 07G3690K), 2 mM L-glutamine (Thermo Fischer, Cat# 25030081), 1mM sodium pyruvate (GIBCO Cat# 11360-039). Hep3B and HepG2 were cultivated in Eagles MEM +
Earle's BSS (PAN, Cat# PO4-08510), 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA (PAN Cat#
P08-32100) and 1 mM sodium pyruvate. JHH2 was cultivated using Williams'E, 10 % FCS
and 2 mM L-glutamine. HLF was cultivated in DMEM 4,5 g/L glucose, 5 % FCS, 0.1 mM
NEAA and 2 mM L-glutamine. SkHep 1 was cultivated in Eagles MEM + Earle's BSS, 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA and 1mM sodium pyruvate. Cells were seeded in the respective media overnight at a density of 5,000 cells per well in 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904).
The next day, logarithmic dilutions of compound 41c-A starting from 27[EM down to 270pM were added and incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 dye (2[Eg/ml, Sigma Cat# B2261). Data shown are means + SD from triplicate wells based on the analysis of 9 images per well relative to the DMSO control.
None of the cell lines tested showed a significant increase in proliferation upon treatment with 41c-A directly after 72 hrs. Results are shown in Figure 6B.
Example 59 Treatment of tumor Cells with an active form of the compounds of the present invention (compound 41c-B) in the presence of peripheral blood results in inhibition of proliferation of tumor cells.
Heparinized whole blood of 3 different donors was diluted 1:1 in RPMI media (PAN Cat.
# PO4-18047) plus 10% FCS (GIBCO Cat# 10500-064, lot 07G3690K) and incubated at 37 C and 5% CO2 for 24hrs with 2.7[EM compound 4 1c-B. Supernatant was harvested and centrifuged at 600x g for 8 minutes to remove residual leukocytes, platelets and erythrocytes. Supernatants were stored at -80 C until further use and thawed gently at room temperature prior to addition to the cell lines. Cell lines Huh7, JHH2, HLE, HLF, JHH4, Hep3B, HepG2, JHH1, EGI1, JHH5, JHH6, OZ, SkHep 1, 5NU449 were seeded in 100[d of the respective media (as described in Example 57) overnight at a density of 5,000 cells per well 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, 100[d of the whole blood supernatants were added to the cell lines. As controls, supernatant of whole blood without addition of 4 lc-B
compound ("whole blood w/o") or plain RPMI media plus FCS ("media CTRL") was added.
Cell lines were incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 (2[Eg/ml, Sigma Cat# B2261) and viability was assessed by additional detection of Propidium Iodine (PI, 1 Kg/ml, Sigma Cat#
P4864).
Data shown are means + SD from triplicate wells based on the analysis of 9 images per well.
Results are shown in Figures 7A and 7B. For some cell lines (SNU449, JHH2 and SkHep) the addition of supernatant of non-stimulated whole blood induced proliferation above the media control level, while others responded with reduced proliferation (OZ, JHH1, HepG2, JHH4, JHH6, JHH5 and EGI1). However, treatment with supernatants derived from whole blood incubated with 4 lc-B resulted in reduced cell counts in all cases tested compared to the respective "whole blood w/o" controls. The reduced cell counts were mainly attributed to a stop in proliferation, and only the cell lines JHH2, JHH4, JHH6, Hep3B
and EGI1 did undergo cell death as determined by considerable PI positivity (data not shown).
Example 60 Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-A) results in inhibition of proliferation in tumor cell lines Heparinized whole blood of 2 different donors was diluted 1:1 in RPMI media (PAN Cat.
# PO4-18047) plus 10% FCS (GIBCO Cat# 10500-064, lot 07G3690K) and incubated at 37 C and 5% CO2 for 24hrs with 2.7[EM compound 4 lc-A. Supernatant was harvested and centrifuged at 600x g for 8 minutes to remove residual leukocytes, platelets and erythrocytes. Supernatants were stored at -80 C until further use and thawed gently at room temperature prior to addition to the cell lines. Cell lines Huh7, JHH2, HLF, Hep3B, HepG2, EGI1 and SkHep 1 were seeded in 100[d of the respective media (as described in Figure 6) overnight at a density of 5,000 cells per well 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, 100 1 of the whole blood supernatants were added to the cell lines. As controls, supernatant of whole blood without addition of 41c-A compound ("whole blood w/o") or plain RPMI
media plus FCS ("media CTRL") was added. Cell lines were incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 (2[Eg/ml, Sigma Cat# B2261) and viability was assessed by additional detection of Propidium Iodine (PI, 1 lag/ml, Sigma Cat# P4864). Data shown are means + SD
from triplicate wells based on the analysis of 9 images per well.
Results are shown in Figure 7C- treatment with supernatants derived from whole blood incubated with 41c-A resulted in reduced or stable cell counts in all cases tested compared to the respective "whole blood w/o" controls. Only in Hep3B, donor #5 supernatant increased proliferation of this cell line, while supernatant from donor #4 had no impact on tumor cell proliferation.
Example 61 Single dose PK study in Male Wister-Han Rats The single dose PK in Male Wister-Han Rats was performed to assess pharmacokinetic properties of tested compounds. Two groups of animals were dosed via Gavage (POE) of the respective compound. Blood samples (approximately 20 [iL) were collected via Jugular vein or an alternate site at 15 mm, 30 min, 1H, 2 h, 4 h, 7 h and 24 h post-dose groups. Blood samples were placed into tubes containing EDTA-K2 anticoagulant and centrifuged at 5000 rpm for 6 min at 4 C to separate plasma from the samples. After centrifugation, the resulting plasma was transferred to clean tubes for bioanalysis of both prodrug and active form on LC/MS/MS. In the groups that prodrug were dosed, the concentration of prodrugs in the plasma samples was under the detection limit. The "tested compound" in Table 8 was used as the internal standard for testing the metabolite (active form) of "dose compound" in vivo. The pharmacokinetic parameters were calculated using non-compartmental module of WinNonlin0 Professional 6.2.
The peak concentration (C.) was recorded directly from experimental observations.
The area under the plasma concentration-time curve (AUCort) was calculated using the linear trapezoidal rule up to the last detectable concentration.
Cmax and AUCo-last are two critical PK parameters related to the in vivo efficacy of the tested compound. Compounds with higher Cmax and AUCo-last will lead to the better in vivo efficacy. Results of PK parameters following oral administration of active forms and competitor compounds are given in Table 7. The PK parameters of prodrugs are tabulated in Table 8.
Following oral administration of prodrugs, the active forms were observed in plasma and therefore tested. The exemplified prodrugs of present invention (Example 41-B, 42-A, 42-B, 43-A, 45-A and 45-B) surprisingly showed much improved Cmax (5-175 folds increase) and AUCo-iast (2.5-56 folds increase) comparing with reference compounds (G59620, S-2 and S-3) and compounds mentioned in present invention (Compound 41c-A, 41c-B and 43e-A) which are all active forms. The results clearly demonstrated the unexpected superiority of prodrugs over active forms on PK parameters which led to better in vivo efficacy.
Table 7. The mean plasma concentration and PK parameters of active forms after 5 mg/kg oral dosing Dose Compound compound 41c-A
Time (h) Mean plasma concentration (nM) 0.25 56.3 9.49 8.89 16.75 0.5 33.2 16.74 9.99 27.48 1 83.4 19.33 10.16 32.33 2 136 24.89 8.40 27.34 4 16.7 47.55 11.54 27.38 8* 9.49 52.72 8.17 18.02 24 ND 4.90 ND 5.60 Cmax (nM) 164 52.72 11.54 32.33 AUCo_iast 316 748 95 242.5 (nM=h) Dose Compound Compound Compound Compound compound 41c-B 43e-A 45e-A 45e-B
Time (h) Mean plasma concentration (nM) 0.25 3.41 12.60 64.6 42.8 0.5 0.75 15.22 80.0 52.2 1 2.04 13.01 58.1 37.6 2 5.46 11.98 42.5 24.2 4 2.52 8.20 77.8 53.9 8* 1.21 6.31 34.6 29 24 ND ND 8.6 5.7 Cmax (nM) 5.46 15.22 80.0 53.9 AUCo_iast 767 568 55.8 77 (nM=h) * 7 hrs for Compound 41c-A, Compound 41c-B and Compound 43e-A
Table 8. PK parameters of prodrugs after 5 mg/kg oral dosing Dose compound Tested compound C. (nM) AUCo-iast (nM=h) Example 41-B Compound 41c-A 1315 3658 Example 42-A Compound 41c-A 1742 4867 Example 42-B Compound 41c-B 956 3148 Example 43-A Compound 43e-A 77 229 Example 45-A Compound 45e-B 922 1914 Example 45-B Compound 45e-A 1436 2619 Example 62 LYSA solubility study LYSA study is used to determine the aqueous solubility of tested compounds.
Samples were prepared in duplicate from 10 mM DMSO stock solution. After evaporation of DMSO with a centrifugal vacuum evaporator, the compounds were dissolved in 0.05 M
phosphate buffer (pH 6.5), stirred for one hour and shaken for two hours.
After one night, the solutions were filtered using a microtiter filter plate. Then the filtrate and its 1/10 dilution were analyzed by HPLC-UV. In addition, a four-point calibration curve was prepared from the 10 mM stock solutions and used for the solubility determination of the compounds. The results were in [tg/mL. In case the percentage of sample measured in solution after evaporation divided by the calculated maximum of sample amount was bigger than 80%, the solubility was reported as bigger than this value.
Results of LYSA were shown in Table 9. It was clear that the solubility of active forms were surprisingly improved by 10 to over 200 folds when converted to various prodrugs.
Table 9. Solubility data of particular compounds LYSA of LYSA of Active Corresponding Active Prodrugs Prodrugs Forms Forms ( g/mL) ( g/mL) Example 1 290 Compound le 21 Example 1-A 315 Compound le-A 56 Example 1-B 200 Compound le-B 50 Example 2 615 Compound le 21 Example 2-A >600 Compound le-B 50 Example 2-B >590 Compound le-A 56 Example 3 240 Compound le 21 Example 4 695 Compound le 21 Example 5 >595 Compound le 21 Example 6 140 Compound le 21 Example 7 615 Compound le 21 Example 8 620 Compound le 21 Example 9 >520 Compound le 21 Example 10 120 Compound le 21 Example 11 >618 Compound le 21 Example 12 120 Compound le 21 Example 13 155 Compound le 21 Example 14 225 Compound le 21 Example 15 405 Compound le 21 Example 16 205 Compound le 21 Example 17 190 Compound le 21 Example 25 >670 Compound le 21 Example 26 >690 Compound le 21 Example 27 >380 Compound le 21 Example 28 695 Compound le 21 Example 29 395 Compound le 21 Example 32 125 Compound le 21 Example 36-A 168 Compound 36g-A 6 Example 36-B 209 Compound 36g-B 11 Example 41-A 260 Compound 41c-B 5 Example 41-B 250 Compound 41c-A 1 Example 42-A 225 Compound 41c-A 1 Example 42-B 335 Compound 41c-B 5 Example 43-A 203 Compound 43e-A 13 Example 43-B 170 Compound 43e-B 13 Example 45 172 Compound 45e 152 Example 45-A >560 Compound 45e-A or 90 or 115 Compound 45e-B
Example 45-B 420 Compound 45e-B 115 or 90 Or Compound 45e-A
Example 46-A 205 Compound 45e-A 90 or 115 Or Compound 45e-B
Example 46-B >580 Compound 45e-B 115 or 90 Or Compound 45e-A
Example 47-A 154 Compound 47e-A or <1.0 or <1.0 Compound 47e-B
Example 47-B 128 Compound 47e-B or <1.0 or <1.0 Compound 47e-A
Example 48-A 305 Compound 47e-A or <1.0 or <1.0 Compound 47e-B
Example 48-B 275 Compound 47e-B or <1.0 or <1.0 Compound 47e-A
Example 63 Portal Vein Study The objective of this study was to understand whether pro drug remains unchanged as it was absorbed through the intestine into the portal circulation and demonstrate the primary site of conversion.
Surgical procedure for Portal vein cannulation (PVC) and Carotid artery cannulation (CAC) Surgery was performed under pentobarbital /isoflurane anesthesia. Briefly, after disinfecting the abdominal area with betadine and 70% isopropyl alcohol, a small abdominal mid-line incision was made. The cecum was pulled out and mesenteric vein was identified and isolated for about 5 mm vessel. A loose ligature was placed proximally and distal end of the vein was ligated. Make a small incision (just enough to allow the insertion of the catheter) on isolated vein and insert the PU catheter towards liver for appropriate length. The catheter was secured in place by tying the loose ligature around the cannulated vessel. The cecum was replaced into abdominal cavity. A hole was made in the right abdominal wall to make the end of catheter pass freely. The catheter was secured by suture on the abdominal wall. The abdominal muscle incision was closed with suture. A
small incision was made in the scapular area to serve as the exit site of the catheter. The catheter was subcutaneously tunneled and exteriorized through the scapular incision. A
fixed suture was placed in the scapular region. The patency of the catheter was checked and then exteriorized from the subcutaneous space to the dorsal neck region. After gently wiping the area, the abdominal cavity was sutured. The left carotid artery was then cannulated by inserting a PESO catheter. Both the exteriorized catheters were tied firmly on the dorsal neck region and fixed. The animals was then allowed to recover in its cage and used for study at least 3 days after surgery. All catheters were flushed once daily with heparinized saline to maintain patency.
Oral PK study in PVC/CAC dual cannulated rat Animals were fasted overnight (n=3) and administered vial oral gavage (10mg/kg, 10mL/kg). Blood samples (60 [LL) were collected simultaneously from the portal and carotid artery catheters at 0.083, 0.25, 0.5, 1, 2, 4, 7, 24h. All blood samples will be transferred into microcentrifuge tubes containing 2 [LI., of K2EDTA (0.5M) as anti-coagulant and placed on wet ice. Then blood samples will be processed for plasma by centrifugation at approximately 4 C, 3000g within half an hour of collection.
Plasma samples will be stored in polypropylene tubes, quick frozen over dry ice and kept at -70 10 C until LC/MS/MS analysis.
Pharmacokinetic parameters (mean SD, n= 3) of prodrugs and active forms in portal and carotid samples following oral administration of prodrugs (10 mg/kg) in portal vein cannulated rat were detected and analyzed. The test results of Example 1-B, 41-A, 41-B, 42-A and 43-A were summarized below.
Table 10. Pharmacokinetic parameters of Example 41-A and its corresponding active form Compound 41c-B in portal and carotid samples following oral administration of Example 41-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 41-A
Corresponding Active Form Compound 41c-B
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.14 0.4 0.19 0.42 C. (nM) 9703 2223 210 2185 AUC0_2 (nM=h) 2188 2246 114 2108 AUCactive/AUC 1 tota, 51% 95%
Table 11. Pharmacokinetic parameters of Example 43-A and its corresponding active form Compound 43e-A in portal and carotid samples following oral administration of Example 43-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 43-A
Corresponding Active Form Compound 43e-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form Tmax (h) 0.28 0.33 0.22 0.28 C. (nM) 4110 818 191 691 AUC0_2 (nM=h) 2067 679 124 564 AUCactive/AUC 1 tota, 25% 82%
Table 12. Pharmacokinetic parameters of Example 1-B and its corresponding active form Compound le-A in portal and systemic samples following oral administration of Example 1-B (10 mg/kg) in portal vein cannulated rat Prodrug Example 1-B
Corresponding Active Form Compound le-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.083 0.25 0.083 0.5 C. (nM) 670 192 70 174 AUC0_2 (nM=h) 266 164 40 184 AUCactive/AUC 1 tota, 38% 82%
Table 13. Pharmacokinetic parameters of Example 42-A and its corresponding active form Compound 41c-A in portal and carotid samples following oral administration of Example 42-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 42-A
Corresponding Active Form Compound 41c-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.19 0.42 0.22 0.36 C. (nM) 8917 3162 286 3326 AUC0_2 (nM=h) 3461 3199 286 3326 AUCactive/AUC 1 tota, 48% 96%
Table 14. Pharmacokinetic parameters of Example 41-B and its corresponding active form Compound 41c-A in portal and carotid samples following oral administration of Example 41-B (10 mg/kg) in portal vein cannulated rat Prodrug Example 41-B
Corresponding Active Form Compound 41c-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form Triax (h) 0.19 0.5 0.25 0.5 C. (nM) 7068 3315 29.6 3432 AUC0_2 (nM=h) 1444 3211 22.5 3301 AUCactive/AUC
tota, 69% 99%
Based on the above results, it was concluded that the primary site of conversion of prodrug was liver rather than intestine, because AUCactive/AUC I was higher in sampling tota, from carotid artery compared to AUCactive/AUCtota, in sampling from portal vein.
H
NN
I
SIN N
*
li A solution of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (58.0 g, Compound 1h) and PMBNH2 (54.7 g, 398.42 mmol) in n-BuOH (600 mL) was stirred at 120 C for 20 hrs. The reaction was concentrated and the residue was washed with MTBE
(400 mL) to give 9-benzy1-6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-7H-purin-8-one (75 g, Compound li) as a white solid and was used in next step without further purification. MS obsd. (ESL') [(M+H)+]: 436.2.
Step 5: Preparation of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c) NJCNH
I 'C3' SN N
#
lc 9-Benzy1-6-[(4-methoxyphenyOmethylamino]-2-propylsulfanyl-7H-purin-8-one (87.0 g, Compound li) in TFA (200 mL) was stirred at 80 C for 16 hrs. The resulting reaction mixture was concentrated, basified with saturated aq. NaHCO3 (600 mL). The resulting precipitate was collected by filtration and washed with (PE/DCM =
2:1, 400mL) to give 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (38.0 g, Compound 1c) as a white solid. MS obsd. (ESL') [(M+H)+]: 316.1.
Step 6: Preparation of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d) N
I
S)N N
id To a solution of m-CPBA(22.98 g, 113.2 mmol) in THF (50 mL) was added dropwise to a suspension of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (35.0 g, compound 1c) in THF (200 mL) at 0 C. After the addition, the reaction mixture was stirred at 25 C for 0.5 hr. The mixture was filtered and washed with MeCN
(400 mL), MTBE (500 mL) to give 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (35.1 g, Compound 1d) as a white solid, which was used for the next step without further purification. MS obsd. (ESL') [(M+H)+]: 332.1.
Step 7: Preparation of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) ON\
H N=S N N
le To a solution of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (34.0 g, Compound 1d) in Eaton's reagent (170.0 mL, 7.5 wt. % in methanesulphonic acid) was added NaN3 (15.34 g, 253.97 mmol) at 60 C slowly. Then the mixture was stirred at 60 C
for 30 mins. The resulting reaction mixture was cooled to 25 C, poured into ice cold NH3H20 (500 mL, 1 mol/L), extracted with n-BuOH (100 mL) four times and concentrated in vacuo. The residue was purified byprep-HPLC to give 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (10 g, Compound le). 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.65 (br. s., 1H), 7.26-7.37 (m, 5H), 6.98 (br. s., 2H), 4.97 (s, 2H), 4.02 (s, 1H), 3.33 (t, J= 7.53 Hz, 2H), 1.55-1.74 (m, 2H), 0.92 (t, J=7.53 Hz, 3H). MS
obsd. (ESE') [(M H)+]: 347.
Example 2 6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NH2 ...-N
T \
N----N
o 1 1 µNs' ..--N
......õ,..õ......õ, µN N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-(2-methoxyethyl)-N-methyl-carbamoyl chloride (Intermediate AB) instead ofN-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (120 mg, Example 2) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
7.27-7.39 (m, 5H), 6.89 (br. s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br. dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J= 8Hz, 3H). MS obsd. (Esr) [(M+H)+]: 462.
Separation of compound of Example 2 by chiral HPLC afforded Example 2-A
(faster eluting, 33 mg) and Example 2-B (slower eluting, 46 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak OJ-3 column.
Example 2-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.89 (br.
s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br.
dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J
= 8Hz, 3H). MS obsd. (ESL') [(M+H)+]: 462.
Example 2-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.89 (br.
s., 1H), 6.78 (br. s., 1H), 5.00 (s, 2H), 4.16 (br. d, J= 4 Hz, 1H), 3.62 (br.
dd,J= 4, 12 Hz, 2H), 3.28-3.42 (m, 6H), 3.12 (d, J= 12 Hz, 3H), 3.05 (s, 1H), 1.58-1.72 (m, 2H), 0.93 (t, J
= 8Hz, 3H). MS obsd. (ESL') [(M+H)+]: 462.
Example 3 6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 r----1 N'\._.....N
9µ j 0 .Sµx -N N
NH
AP
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-propyl-carbamoyl chloride (Intermediate AC) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (51 mg, Example 3) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.85 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. d, J= 8.0 Hz, 1H), 3.13-3.54 (m, 4H), 1.46-1.72 (m, 4H), 1.30-1.39 (m, 1H), 1.00-1.26 (m, 6H), 0.81-0.95 (m, 5H), 0.73 (t, J= 8 Hz, 1H). MS
obsd. (ESL') [(M+H)+]: 474.
Example 4 6-Amino-9-benzy1-7-14-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one N
NJ
//Sµµr\JH N "
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using (1,4'-bipiperidine)-1'-carbonyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one (55 mg, Example 4) was obtained as a white powder.
- 114 -11-1 NMR (400 MHz, DMSO-d6) 6 ppm: 7.39 - 7.27 (m, 5H), 6.97 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. s., 2H), 3.85 (d, J= 12.5 Hz, 1H), 3.43 - 3.15 (m, 3H), 2.96 (t, J=
12.3 Hz, 2H), 2.56 (m, 4H), 1.83 (m, 1H), 1.79 - 1.54 (m, 4H), 1.50 (br. s., 4H), 1.45 -1.33 (m, 3H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 541.2.
Example 5 6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyDpurine-7-carboxamide 0 rj \---Nj----N
µµS
N 1\1 ---NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (Intermediate AD) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (34 mg, Example 5) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.39 -7.28 (m, 5H), 6.89 (br. s., 1H), 6.74 (br. s., 1H), 4.99 (s, 2H), 4.17 (d, J= 8.1 Hz, 1H), 3.67 (br. s., 2H), 3.63 - 3.51 (m, 2H), 3.50 - 3.34 (m, 4H), 3.29 (s, 1H), 3.11 (s, 2H), 1.73- 1.59 (m, 2H), 1.23 - 1.07 (m, 3H), 0.93 (t, J= 7.5 Hz, 3H). MS obsd. (Esr) [(M+H)+]:
476.3.
Example 6 6-Amino-9-benzyl-N-butyl-N-ethyl-8-oxo-2-(propylsulfonimidoyDpurine-7-carboxamide 0 ri-Nj\.___N
9µ 0 //Sµµ N N
NH
At The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-butyl-N-ethyl-carbamoyl chloride (Intermediate AE) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-butyl-N-ethy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (51 mg, Example 6) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.85 (br. s., 2H), 4.99 (s, 2H), 4.20 (br. d, J= 8.0 Hz, 1H), 3.13-3.54 (m, 4H), 1.46-1.72 (m, 4H), 1.30-1.39 (m, 1H), 1.00-1.26 (m, 6H), 0.81-0.95 (m, 5H), 0.73 (t, J= 8 Hz, 1H). MS obsd.
(ESL') [(M+H)+]: 474.
Example 7 6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NH2 .......N
Ni\li \\
9µ )=0 Sµµ N N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-(2-methoxyethyl)carbamoyl chloride (Intermediate AF) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide (35 mg, Example 7) was obtained as a white powder. 'H NMR (400 MHz, DMSO-d6) 6 ppm:
7.40 - 7.28 (m, 5H), 6.89 (br. s., 1H), 6.75 (br. s., 1H), 5.00 (d, J= 5.5 Hz, 2H), 4.24 - 4.16 (m, 1H), 3.77 (br. s., 1H), 3.67 (br. s., 1H), 3.62 - 3.53 (m, 1H), 3.42 - 3.27 (m, 5H), 3.23 -3.02 (m, 3H), 1.66-1.38 (m, 4H), 0.96 - 0.70 (m, 6H). MS obsd. (ESL') [(M+H)+]: 490.5.
Example 8 6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide 0 rj NJ.---9µ j 0 SN N N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using bis(2-methoxyethyl)carbamic chloride (Intermediate AG) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (35 mg, Example 8) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.40 - 7.28 (m, 5H), 6.83 (br. s., 2H), 4.99 (s, 2H), 3.71 (br. s., 3H), 3.52 - 3.27 (m, 11H), 3.09 (s, 3H), 1.73- 1.59 (m, 2H), 0.93 (t, J= 7.5 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 506.
Example 9 6-Amino-7-(azetidine-1-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one NN
OµN j I
S' N' N
NH
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using azetidine-l-carbonyl chloride (Intermediate AH) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-7-(azetidine-1-carb ony1)-9-b enzy1-2-(propylsulfonimidoyl)purin-8-one (120 mg, Example 9) was obtained as a white powder.
1HNMR (400 MHz, DMSO-d6) 6 ppm: 7.02 - 7.43 (m, 7H), 4.99 (s, 2H), 4.31 (t, J=
7.65 Hz, 2H), 4.08 - 4.23 (m, 3H), 3.34 - 3.41 (m, 2H), 2.28 (m, 2H), 1.56 - 1.73 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 430.
Example 10 6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2 ......1\1 / \
N----N
µµS N
/..\,=-=' µµ N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 10 using N-isopropyl-N-methyl-carbamoyl chloride (Intermediate Al) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (97 mg, Example 10) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.39 (m, 5H), 6.87 (br. s., 2H), 4.99 (s, 2H), 4.38-4.45 (m, 1H), 4.09-4.21 (m, 1H), 3.29-3.43 (m, 2H), 2.89-2.95 (m, 3H), 1.58-1.73 (m, 2H), 1.21 (br d, J= 8 Hz, 6H), 0.93 (t, J= 8 Hz, 3H). MS obsd.
(ESL') [(M+H)+]: 446.
Example 11 6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one 0 r--\ N---NH2 ...... N \..... j N----N
0µµ I
S' N "
NH
*
ii The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 4-methylpiperazine-1-carbonyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzy1-7-(4-methylpiperazine-1-carbony1)-(propylsulfonimidoyl)purin-8-one (59.5 mg, Example 11) was obtained as a yellow solid.
1H NMR (400 MHz, DMSO-d6) gppm: 7.39 - 7.31 (m, 5H), 6.99 (s, 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.58 - 3.49 (m, 6H), 2.42 (m, 4H), 2.22 (s, 3H), 1.66 - 1.61 (m, 2H), 0.95 -0.91 (t, J= 7.2 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 473.
Example 12 6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide /
0 ri NH2 ...-N
J \
N-----N
o1Io "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-(3-methoxypropy1)-N-methyl-carbamoyl chloride instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (92.2 mg, Example 12) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.23 - 7.45 (m, 5H), 6.94 (s., 2H), 4.93-5.08 (m, 2H), 4.19 (s, 1H), 3.30 - 3.62 (m, 6H), 3.25 (s, 3H), 3.02 - 3.10 (m, 3H), 1.74 - 1.90 (m, 2H), 1.55 - 1.77 (m, 2H), 0.98 - 0.82 (m, 3H). MS obsd.
(ESL') [(M+H)+]: 476.3.
Example 13 6-Amino-9-benzyl-N-isobutyl-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2 ...-N
\
N
N.---µµ ...---N
//Sµ\NH N
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-isobutyl-N-methyl-carbamoyl chloride (Intermediate AL) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-benzyl-N-isobutyl-N-methyl-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (64 mg, Example 13) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.27-7.40 (m, 5H), 6.89 (br. s., 2H), 5.00 (s, 2H), 4.16 (br. s., 1H), 3.25-3.44 (m, 4H), 3.07 (s, 2H), 3.03 (s, 1H), 1.87-2.09 (m, 1H), 1.57-1.74 (m, 2H), 0.75-0.99 (m, 9H). MS obsd. (ESL') [(M+H)+]: 460.
Example 14 Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]acetate 0---j 0 r40 NH2 ..-N
1 \
N.......N
ON\ IIo "
\µ N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl 2-((chlorocarbonyl)(methyl)amino)acetate (Intermediate AP) instead ofN-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl 24[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] acetate (38 mg, Example 14) was obtained as a light yellow powder. 1H NMR (400MHz, DMSO-d6) 6 ppm:
7.41 -7.27 (m, 5H), 6.82 (br. s., 1H), 5.04 - 4.95 (m, 2H), 4.35 (br. s., 1H), 4.28 (br. s., 1H), 4.23 - 4.16 (m, 2H), 4.08 (q, J= 7.2 Hz, 1H), 3.43 - 3.28 (m, 3H), 3.15 (s, 2H), 3.08 (s, 1H), 1.71 - 1.58 (m, 2H), 1.24 (t, J= 7.0 Hz, 2H), 1.12 (t, J= 7.0 Hz, 1H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 490.
Example 15 Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate 0 r r ,..... 0 NNII \
(:)µµ I
µ\ N "
NH
ilt 15 The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl 3-((chlorocarbonyl)(methyl)amino)propanoate instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate (35 mg, Example 15) was obtained as a white powder. 'H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.26 (m, 5H), 6.93 (br. s., 2H), 4.99 (s, 2H), 4.16 (s, 1H), 4.08 (q, J= 7.1 Hz, 1H), 3.99 (d, J= 7.0 Hz, 1H), 3.67 (br. s., 2H), 3.40 - 3.29 (m, 2H), 3.08 (s, 2H), 2.99 (s, 1H), 2.71 (t, J= 6.4 Hz, 2H), 1.74- 1.56 (m, 2H), 1.27- 1.05 (m, 3H), 0.93 (t, J= 7.5 Hz, 3H). MS
obsd.
(ES[) [(M+H)+]: 504.
Example 16 tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate 0 \/----ry,0 \
N
N.--µN -----N1 N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl 3-[chlorocarbonyl(methyl)amino]propanoate (Intermediate AR) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate (60 mg, Example 16) was obtained as a white powder. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
7.41 - 7.27 (m, 5H), 6.93 (br. s., 2H), 4.99 (s, 2H), 4.15 (s, 1H), 3.64 (br.
s., 2H), 3.51 -3.33 (m, 2H), 3.08 (s, 2H), 2.98 (s, 1H), 2.62 (t, J= 6.9 Hz, 2H), 1.71 - 1.57 (m, 2H), 1.41 (s, 6H), 1.34 (s, 3H), 0.93 (t, J= 7.4 Hz, 3H). MS obsd. (ES[) [(M+H)+]: 532.
Example 17 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate \
Nj\____N
oNN
SNµ N
NH N
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (2S)-2-[chlorocarbonyl(methyl)amino]propanoate (Intermediate AS) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate (34.1 mg, Example 17) was obtained as a yellow solid. 1H NMR (300 MHz, DMSO-d6) 6 ppm:
7.22 - 7.49 (m, 5 H), 6.78 (br. s., 2H), 4.93 - 5.08 (m, 2H), 4.75 (br. s., 1H), 3.96 - 4.29 (m, 3H), 3.30 - 3.46 (m, 2H), 3.09 (s, 2H), 2.93 (br. s., 1H), 1.55- 1.77 (m, 2H), 1.48 (d, J=
7.16 Hz, 3H), 1.09 - 1.29 (m, 3H), 0.94 (t, J = 7.44 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
504.2.
Example 18 tert-Butyl (2S)-2- I [6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl] -methyl-amino] -4-methyl-pe ntano ate _........4)--\
Nj\õ.....N
.
oxµ 0 "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AT) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (22 mg, Example 18) was obtained as a white solid.
1H NMR (400MHz, DMSO-d6) 6 ppm: 7.42 - 7.27 (m, 5H), 6.78 (br. s., 2H), 5.05 -4.96 (m, 2H), 4.78 (br. s., 1H), 4.33 (br. s., 1H), 3.51 - 3.37 (m, 2H), 3.01 (s, 3H), 1.75 - 1.54 (m, 4H), 1.44 (s, 8H), 1.33- 1.11 (m, 2H), 0.99 - 0.82 (m, 9H). MS obsd.
(ESL') [(M+H)+]:
574.3.
Example 19 Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]-4-methyl-pentanoate o 0 \
Nj\,....N
0 I I .
µNS N .....õ,........õ,/, µN
NH
N
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using isopropyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AU) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (43 mg, Example 19) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.27 (m, 5H), 6.75 (br. s., 2H), 5.05 - 4.94 (m, 3H), 4.88 (br. s., 1H), 4.19 (br. s., 1H), 3.43 - 3.34 (m, 2H), 3.01 (s, 3H), 1.91 (br. s., 1H), 1.77 - 1.56 (m, 4H), 1.25 - 1.16 (m, 6H), 0.99 - 0.83 (m, 9H).
MS obsd. (ESC') [(M+H)+]: 560.3.
Example 20 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate 0¨i o-----40 NH2 ...-N
\
N
N--9µ I
S\\ N "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-methyl-butanoate (Intermediate AV) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-5 amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate (51.5 mg, Example 20) was obtained as a white powder. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.23 - 7.51 (m, 5H), 6.76 (br. s., 2H), 5.01 (br. s., 2H), 4.42 (br. s., 1H), 3.97 - 4.26 (m, 3H), 3.34 - 3.45 (m, 2H), 3.12 (br. s., 3H), 2.24 (br.
s., 1H), 1.65 (br. s., 2H), 1.13 - 1.29 (m, 3H), 0.88 - 1.10 (m, 9H). MS obsd. (ESI+) [M+H ]: 532.2.
10 Example 21 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate J
0 ......:40 NH2 ..-N
\
N
N--9µ I
Sµµ N "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 15 using ethyl (25)-2-[chlorocarbonyl(methyl)amino]-4-methyl-pentanoate (Intermediate AW) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate (17.3 mg, Example 21) was obtained as a white powder. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.26 - 7.45 (m, 5H), 6.73 (br. s., 2H), 4.91 - 5.09 (m, 3H), 4.06 -4.25 (m, 3H), 3.34 - 3.45 (m, 2H), 3.04 (br. s., 3H), 1.93 (br. s., 1H), 1.54 -1.78 (m, 4H), 1.22 (t, J= 7.09 Hz, 3H), 0.77 - 1.01 (m, 9H). MS obsd. (ES[) [(M+H)+]: 546.3.
Example 22 Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]-3-phenyl-propanoate NH2 ...-N
\
N\ õ...... N
N
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl (25)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AX) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (30 mg, Example 22) was obtained as a white powder. 1H NMR
(400MHz, DMSO-d6) 6 ppm: 7.42 - 7.16 (m, 10H), 4.97 (s, 3H), 4.19 (q, J= 7.1 Hz, 2H), 3.35 -3.15 (m, 6H), 3.10 - 2.90 (m, 3H), 1.71 - 1.46 (m, 2H), 1.28- 1.18 (m, 4H), 0.97 -0.85 (m, 3H). MS obsd. (ES[) [(M+H)+]: 580.
Example 23 Isopropyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate 4Ik 0----( \
Nj\,....N
oNN .
\µ N
NH
"
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using isopropyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AY) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (22 mg, Example 23) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.35 - 7.01 (m, 10H), 5.02-4.89 (m, 3H), 3.37-3.17 (m, 3H), 3.02 - 3.09 (m, 3H), 3.10 -2.90 (m, 3H), 1.66 - 1.62 (m, 2H), 1.22 -1.11 (m, 8H), 0.92 (t, J= 7.4 Hz, 3H). MS obsd. (ESI+) [(M+H)+]: 594.
Example 24 tert-Butyl (2S)-2-[16-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate 4Ik 0----k-N--- NI \
% 1.
-N N
NH
4111\
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl (2S)-2-[chlorocarbonyl(methyl)amino]-3-phenyl-propanoate (Intermediate AZ) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl (2,5)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate (34 mg, Example 24) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.42 - 7.16 (m, 10H), 5.03 - 4.90 (m, 3H), 3.68 - 3.24 (m, 5H), 3.24 - 3.09 (m, 2H), 3.01 (s, 3H), 1.68 - 1.57 (m, 2H), 1.43 (s, 9H), 0.99 - 0.85 (m, 3H). MS obsd. (ESL') [(M+H)+]: 608.3.
Example 25 N- 12- [Acetyl(methyl)amino] ethyl] -6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide \ N
NH2 ...-N
1 \
N'N
ID\ I
S
-----N
µ N
...õ..."--.........õ., µµ
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N- [2-[acetyl(methyl)amino]ethyl]-N-methyl-carbamoyl chloride (Intermediate BA) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). N-[2-15 [Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (26.1 mg, Example 25) was obtained as a white powder.1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.27 (m, 5H), 7.02 (br, 2H), 5.04 - 4.97 (m, 2H), 4.19 - 4.13 (m, 1H), 3.57 (d, J= 5.5 Hz, 2H), 3.49 - 3.34 (m, 2H), 3.14 (s, 1H), 3.12 - 3.02 (m, 4H), 2.86 (d, J= 7.5 Hz, 2H), 2.69 - 2.64 (m, 1H), 2.05 (s, 1H), 20 1.99 (s, 1H), 1.91 - 1.83 (m, 1H), 1.70 - 1.59 (m, 2H), 0.97 - 0.90 (m, 3H). MS obsd.
(ESI ) [(M+H)+]: 503.2.
Example 26 Methyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino] ethyl] -N-methyl-c arbam ate \ N40 0 r---1 NH2 ......N
\
N
N).--% I
Sµµr\jH N "
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using methyl N42-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (Intermediate BB) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Methyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (65 mg, Example 26) was obtained as a yellow solid. 1H NMR (400 MHz, CDC13) 6 ppm: 7.29 - 7.49 (m, 5H), 5.63 - 5.92 (m, 2H), 5.03 -5.17 (m, 2H), 3.43 - 3.69 (m, 8H), 3.13 - 3.27 (m, 3H), 2.96 - 3.05 (m, 2H), 2.72 (br. s., 1H), 1.05 (t, J= 7.40 Hz, 3H), 1.87 (dd, J= 14.12, 6.96 Hz, 2H). MS obsd.
(ES[) [(M+H)+]: 519.2.
Example 27 tert-Butyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino] ethyl] -N-methyl-c arbam ate O---k \ N40 0 ri \
N
N
ONN 1 )=C) µ N "
'NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using tert-butyl N- [2-[chlorocarbonyl(methyl)amino]ethyl]-N-methyl-carbamate (Intermediate BC) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
tert-Butyl N- [2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (32 mg, Example 27) was obtained as a white powder. 1H NMR (400MHz, DMSO-d6) 6 ppm: 7.43 - 7.26 (m, 5H), 6.89 (br. s., 2H), 4.99 (d, J= 5.0 Hz, 2H), 4.16 (s, 1H), 3.55 (br. s., 2H), 3.48 - 3.34 (m, 2H), 3.10 (s, 2H), 3.07 (s, 1H), 2.86 (d, J= 12.8 Hz, 2H), 2.74 (d, J= 9.5 Hz, 1H), 2.70 - 2.60 (m, 1H), 1.72 - 1.54 (m, 2H), 1.39 (s, 6H), 1.23 (s, 2H), 1.13 (s, 2H), 0.93 (t, J= 7.4 Hz, 3H). MS
obsd. (Esr) [(M+H)+]: 562.
Example 28 Ethyl N- 12-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino] ethyl] -N-methyl-carbamate \ N40 0 ri NH2 ...-N
N---Nil \
oµµ j 0 -N N
*15 "NH 28 The title compound was prepared in analogy to Example 1, Method A, Step 6 by using ethyl N42-[chlorocarbonyl(methyl)amino]ethy1]-N-methyl-carbamate (Intermediate BD) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). Ethyl N-[2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate (87 mg, Example 28) was obtained as a yellow solid.1H
NMR (400 MHz, CDC13) 6 ppm: 7.29 - 7.53 (m, 5H), 5.65 - 5.90 (m, 2H), 5.02 -5.14 (m, 2H), 3.38 - 4.21 (m, 9H), 3.14 - 3.26 (m, 3H), 3.00 (br. s., 2H), 2.73 (s, 1H), 1.76 - 1.99 (m, 2H), 1.22 - 1.31 (m, 3H), 1.05 (s, 3H). MS obsd. (ESE') [(M+H)+]: 533.2.
Example 29 2-116-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyli-methyl-amino] ethyl N-butyl-N-methyl-carbamate N
NH2 ...-N
j \
µµ N S N
............õ µµ
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl N-butyl-N-methyl-carbamate (Intermediate BE) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl N-butyl-N-methyl-carbamate (19 mg, Compound 29) was obtained as yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.25 - 7.48 (m, 5H), 6.96 (br. s., 2H), 4.99 (s, 2H), 4.06 - 4.36 (m, 3H), 3.59 - 3.83 (m, 1H), 3.33 - 3.49 (m, 3H), 3.07 - 3.21 (m, 4H), 2.79 (s, 2H), 1.65 (br. s., 2H), 1.05 - 1.47 (m, 6H), 0.93 (t, J= 7.40 Hz, 3H), 0.70 - 0.87 (m, 3H). MS
obsd. (ESE) [(M+H)+]: 561.2.
Example 30 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl pyrrolidine-l-carboxylate Cs N----0 rj NH2 ....- N
\
Nj\,....N
O I
N\
/\Sµµ " "
NH
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 5 using 2-[chlorocarbonyl(methyl)amino]ethyl pyrrolidine-l-carboxylate (Intermediate BF) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl pyrrolidine-l-carboxylate (10.0 mg, Example 30) was obtained as a yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.26 - 7.41 (m, 5H), 6.96 (br.s., 2H), 4.99 (s, 2H), 4.01 -4.35 (m, 4H), 10 3.29 - 3.47 (m, 3H), 3.23 (br. s., 3H), 3.03 - 3.17 (m, 4H), 1.52 - 1.84 (m, 6H), 0.90 ¨0.96 (m, 3H). MS obsd. (ESI+) [(M+H)+]: 545.2.
Example 31 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyli-methyl-aminoiethyl N-methyl-N-propyl-carbamate \ NX
0 ri NH2 ...-N
\
NN
9µ 0 N
S\µ N
NH
=
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl N-methyl-N-propyl-carbamate (Intermediate BG) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-methyl-N-propyl-carbamate (3.7 mg, Example 31) was obtained as a yellow solid. 1H NMR
(400 MHz, CD30D) 6 ppm: 7.22 - 7.48 (m, 5H), 5.09 - 5.22 (m, 4H), 4.55 (s, 2H), 3.38 - 3.57 (m, 4H), 3.13 (s, 3H), 1.61 - 1.85 (m, 4H), 1.22- 1.41 (m, 3H), 0.88 - 1.13 (m, 6H). MS
obsd. (ESI+) [(M+H)+]: 547.2.
Example 32 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N,N-diethylcarbamate NJN
0 rj NH2 ....-N
T \
Nj\_....N
9µ )=0 "
\µ N
NH
*
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl /V,N-diethylcarbamate (Intermediate BH) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 24[6-Amino-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl N,N-diethylcarbamate (21.7 mg, Example 32) was obtained as yellow solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.25 - 7.41 (m, 5H), 6.96 (br. s., 2H), 4.99 (s, 2H), 4.08 - 4.36 (m, 3H), 3.70 (br, 1H), 3.33 - 3.46 (m, 3H), 3.01 - 3.24 (m, 7H), 1.55 - 1.74 (m, 2H), 0.86 -1.05 (m, 9H). MS obsd. (ESL') [(M+H)+]: 547.2.
Example 33 2-116-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl ethyl carbonate 0-i 0 ri NH2 ...-N
NN1 \
N
NH
.
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 2-[chlorocarbonyl(methyl)amino]ethyl ethyl carbonate (Intermediate BI) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 2-[[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethyl ethyl carbonate (46 mg, Example 33) was obtained as yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
0.82 -0.99 (m, 3H), 1.02 - 1.28 (m, 3H), 1.56 - 1.76 (m, 2H), 3.05 - 3.18 (m, 3H), 3.35 - 3.48 (m, 3H), 3.73 (t, J= 5.08 Hz, 2H), 4.08 - 4.27 (m, 3H), 4.37 (br. s., 1H), 5.00 (s, 2H), 6.76 -7.11 (m, 2H), 7.22 - 7.45 (m, 5H). MS obsd. (ESE') [(M+H)+]: 520.
Example 34-A and Example 34-B
6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-IS(S)-propylsulfonimidoyl]purine-7-carboxamide and 6-amino-N-buty1-9-1(4-chlorophenyllmethyli-N-methyl-8-oxo-2-1S(S)-propylsulfonimidoylipurine-7-carboxamide r r NitoyN NH20N
N\...õ, N
NJ---N
cNIN s ,, )=0 H NNN .0L I )=0 ' N N , S N N
I-1 NV 0' = CI . CI
Step 1: Preparation of 4-amino-3-[(4-chlorophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 34a) N.........H
N
1 ) _____________________________________ 0 34a Compound 34a was prepared in analogy to Example 1, Method A, Step 1 by using 4-chlorobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-chlorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (8.0 g, Compound 34a) was obtained as a yellow solid. MS obsd. (ESI+) [(M+H)+]: 249.
Step 2: Preparation of 6-amino-9-[(4-chlorophenyl)methyl]-2-sulfany1-7H-purin-one (Compound 34b) H
N-----"N
1 ) __ 0 H S-1\1.-----N
34h Compound 34b was prepared in analogy to Example 1, Method A, Step 2 by using 4-Amino-3-[(4-chlorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 34a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la).
6-Amino-9-[(4-chlorophenyl)methy1]-2-sulfany1-7H-purin-8-one (6.4 g, Compound 34b) was obtained as a yellow solid and was used for the next step without further purification.
MS obsd. (ESL') [(M+H)+]: 308.
Step 3: Preparation of 6-amino-9-[(4-chlorophenyhmethy1]-2-propylsulfany1-7H-purin-8-one (Compound 34c) H
N.------N
1 > __ 0 =SNN
= CI
34c Compound 34c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-chlorophenyOmethy1]-2-sulfany1-7H-purin-8-one (Compound 34b) instead of 6-amino-9-phenylmethy1-2-sulfany1-7H-purin-8-one (Compound lb). 6-Amino-9-[(4-chlorophenyOmethy1]-2-propylsulfany1-7H-purin-8-one (800 mg, Compound 34c) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 350.
Step 4: Preparation of 6-amino-9-[(4-chlorophenyhmethy1]-2-propylsulfiny1-7H-purin-8-one (Compound 34d) H
N-------N
1 ) __ 0 SNN
II
34d Compound 34d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-propylsulfany1-7H-purin-8-one (Compound 34c) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-9-[(4-chlorophenyl)methy1]-2-propylsulfiny1-7H-purin-8-one (150 mg, Compound 34d) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 366.
Step 5: Preparation of 6-amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (compound 34e), 6-amino-9-[(4-chlorophenyl)methy1]-2-1S(S)-propylsulfonimidoy1)-7H-purin-8-one and 6-amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-propylsulfonimidoy1)-7H-purin-8-one (Compound 34e-A and Compound 34e-B) H
N----N
I-1 N\\ I > __ 0=S NN
H = CI
34e H
N-----N
NN H N 1 >
--- _______________________________________________________________ 1 0 1 > ___ \\ ,L
\\ 0.-<.. 0=S 's N'---N
H N=S 's N--N
= CI
= CI
34e-A and 34e-B
Compound 34e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-propylsulfiny1-7H-purin-8-one (Compound 34d) instead of 6-amino-9-benzy1-2-(2-propylsulfiny1)-7H-purin-8-one (Compound 1d).
Amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (250 mg, compound 34e) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm:
10.60 (br. s, 1H), 7.32-7.42 (m, 4H), 6.98 (br. s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.25-3.41 (m, 2H), 1.56-1.68 (m, 2H), 0.91 (t, J= 8 Hz, 3H). MS obsd. (ESL') [(M+1-1) ]:
381.
Separation of compound of Compound 34e by chiral HPLC afforded Compound 34e-A (faster eluting, 110 mg) and Compound 34e-B (slower eluting, 100 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak OJ-3 column.
Compound 34e-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.63 (br. s, 1H), 7.33-7.42 (m, 4H), 6.99 (br. s, 2H), 4.96 (s, 2H), 4.05 (br. s, 1H), 3.26-3.39 (m, 2H), 1.53-1.69 (m, 2H), 0.91 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 381.
Compound 34e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.63 (br. s, 1H), 7.33-7.42 (m, 4H), 6.99 (br. s, 2H), 4.96 (s, 2H), 4.05 (br. s, 1H), 3.26-3.40 (m, 2H), 1.54-1.69 (m, 2H), 0.91 (t, J= 7.5 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 381.
Step 6: 6-Amino-N-buty1-9-[(4-chlorophenyl)methyll-N-methy1-8-oxo-2-1S(S)-propylsulfonimidoyl]purine-7-carboxamide and 6-amino-N-buty1-9-[(4-chlorophenyl)methyll-N-methy1-8-oxo-2-1S(S)-propylsulfonimidoyl]purine-7-carboxamide (Example 34-A and Example 34-B) r NI-I21/4j, N NH2L*N
N N N.....,..N
-' oNN ,, 0 I-1 NµN s, I )=0 ,S ' N N ,S
I-1 NV 0' = CI IIIP CI
Example 34-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 34e-A and N-butyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound 1e) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 34-A (160 mg): 'H NMR (400 MHz, DMSO-d6) 6 ppm: 7.37-7.45 (m, 4H), 6.91 (br. s., 2H), 4.99 (s, 2H), 4.17 (s, 1H), 3.28-3.40 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 1.49-1.70 (m, 4H), 1.15-1.37 (m, 2H), 0.89-0.94 (m, 5H), 0.76 (t, J= 8 Hz, 1H). MS
obsd. (ESL') [(M+H)+]: 494.
Example 34-B (167 mg) was prepared in analogy to Example 34-A by using Compound 34e-B instead of Compound 34e-A.
Example 34-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.36-7.45 (m, 4H), 6.91 (br.
s., 2H), 4.99 (s, 2H), 4.17 (s, 1H), 3.28-3.41 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 1.50-1.71 (m, 4H), 1.15-1.37 (m, 2H), 0.89-0.94 (m, 5H), 0.76 (t, J= 7.4 Hz, 1H). MS obsd.
(ESE) [(M+H)+]: 494.
Example 35 6-Amino-9-[(4-chlorophenyl)methyli-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide NH2uyN
N
N--oNN 0 S N N
I-INV
. CI
The title compound was prepared in analogy to Example 1, Method A, Step 6 by 20 using 6-amino-9-[(4-chlorophenyl)methy1]-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound 34e) and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide (60 mg, Example 35) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.40 (s, 4H), 6.91 (br s, 2H), 4.99 (s, 2H), 4.16 (s, 1H), 3.34-3.44 (m, 4H), 3.05 (s, 2H), 3.01 (s, 1H), 1.58-1.67 (m, 2H), 1.18 (t, J= 8.0 Hz, 3H), 0.92 (t, J= 8.0 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 466.
Example 36-A and Example 36-B
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy11-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide N--N N--'N
I-1 NN, .s,I o s, Nµ 0 ,S N N S ' NJ N
0' . I-1 NV
Step 1: Preparation of 6-chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (Compound 36a) CI
Ni S)N 1 N H
36a Compound 36a was prepared in analogy to Example 1, Method B, Step 1 by using p-tolylmethylamine instead of phenylmethanamine. 6-Chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (3.9 g, Compound 36a) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 353.
STEP 2: PREPARATION OF 6-CHLOR0-2-PROPYLSULFANYL-N4-(P-TOLYLMETHYL)PYRIMIDINE-4,5-DIAMINE (COMPOUND 36B) CI
S(I
N NH
36b Compound 36b was prepared in analogy to Example 1, Method B, Step 2 by using 6-chloro-5-nitro-2-propylsulfanyl-N-(p-tolylmethyl)pyrimidin-4-amine (Compound 36a) instead of N-benzy1-6-chloro-5-nitro-2-propylsulfanyl-pyrimidin-4-amine (Compound if). 6-Chloro-2-propylsulfanyl-N4-(p-tolylmethyl)pyrimidine-4,5-diamine (2.2 g, Compound 36b) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]:
323.
STEP 3: PREPARATION OF 6-CHLOR0-2-PROPYLSULFANYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36C) CI
H
NJN
I
SIN N
36c Compound 36c was prepared in analogy to Example 1, Method B, Step 3 by using 6-chloro-2-propylsulfanyl-N4-(p-tolylmethyl)pyrimidine-4,5-diamine (Compound 36b) instead of N-benzy1-6-chloro-2-(propylsulfanyl)pyrimidine-4,5-diamine (Compound 1g).
6-Chloro-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (2.2 g, Compound 36c) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 349.
STEP 4: PREPARATION OF 6-1-(4-METHOXYPHENYL)METHYLAMIN0]-2-PROPYLSULFANYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36D) NHPMB
H
NN
I
N
36d Compound 36d was prepared in analogy to Example 1, Method B, Step 4, by using 6-chloro-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36c) instead of 9-benzy1-6-chloro-2-propylsulfany1-7H-purin-8-one (Compound 1h). 6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (2.0 g, Compound 36d) was obtained as a white solid. MS obsd. (ESE) [(M+H)+]: 450.
STEP 5: PREPARATION OF 6-AMINO-2-PROPYLSULFAIVYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36E) N\_ S)N N
36e Compound 36e was prepared in analogy to Example 1, Method B, Step 5 by using 6-[(4-methoxyphenyl)methylamino]-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36d) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound ii). 6-amino-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (1.0 g, Compound 36e) was obtained as a white solid. MS obsd. (ESE) [(M+H)+]: 330.
STEP 6: PREPARATION OF 6-AMINO-2-PROPYLSULFINYL-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36F) H
I
N
36f Compound 361 was prepared in analogy to Example 1, Method B, Step 6 by using 6-amino-2-propylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36e) instead of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound 1c). 6-amino-2-propylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (220 mg, Compound 36f) was obtained as a white solid MS obsd. (ESL') [(M+H)+]: 345.
STEP 7: PREPARATION OF 6-AMINO-2-(PROPYLSULFONIMIDOYL)-9-(P-TOLYLMETHYL)-7H-PURIN-8-ONE (COMPOUND 36G) NN
0 >
\\
H N=S N
36g Compound 36g was prepared in analogy to Example 1, Method B, Step 7 by using 6-amino-2-propylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 361) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (127 mg, Compound 36g) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.67 (br. s., 1H), 7.23 (d, J= 8.0 Hz, 2H), 7.13 (d, J= 8.0 Hz, 2H), 6.98 (br. s., 2H), 4.91 (s, 2H), 4.05 (s, 1H), 3.34-3.27 (m, 2H), 2.26 (s, 3H), 1.67-1.62 (m, 2H), 0.92 (t, J= 8.0 Hz, 3H).
MS obsd.
(ESL') [(M+H)+]: 361.
Separation of compound 36g by chiral HPLC afforded compound 36g-A (faster eluting, 50 mg) and compound 36g-B (slower eluting, 49 mg) as white solid with 30%
isopropanol (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 36g-A: 111 NMR: (400 MHz, DMSO-d6) 6 ppm: 10.51 (s, 1 H), 7.22 (d, J=
8.0 Hz, 2H), 7.12 (d, J= 8.0 Hz, 2H), 7.00 (s, 2 H), 4.91 (s, 2H), 4.03 (s, 1H), 3.35 - 3.31 (m, 2H), 2.26 (s, 3H), 1.70 - 1.58 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd.
(ESI ) [(M+H)+]: 361.
Compound 36g-B: 111 NMR: (400 MHz, DMSO-d6) 6 ppm: 10.54 (s, 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J= 8.0 Hz, 2H), 6.97 (s, 2H), 4.91 (s, 2H), 4.04 (s, 1H), 3.34 - 3.30 (m, 2H), 2.26 (s, 3H), 1.72 - 1.57 (m, 2H), 0.93 (t, J= 7.40 Hz, 3H). MS obsd.
(ESL') [(M+H)+]: 361.
Step 8: Preparation of 6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-methyl-8-oxo-N-propy1-2[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide (Example 36-A and Example 36-B) , , NH2L'yN NH-'N
NCN
H Nµµ ,µ I 0 (3µ L
,\S's% N N
I¨IN' 0' 0 *
Example 36-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). Example 36-A (108 mg) was obtained as a white solid. 1H NMR
(400 MHz, DMSO-d6) 6 ppm: 7.27 (d, J= 8 Hz, 2H), 7.14 (d, J= 8 Hz, 2H), 6.87 (br.
s., 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.33-3.57 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 2.26 (s, 3H), 1.52-1.73 (m, 4H), 0.75-0.97 (m, 6H). MS obsd. (ESL') [(M+H)+]: 460.
Example 36-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (compound le). Example 36-B(125 mg): 1H NMR (400 MHz, DMSO-d6) 5 ppm: 7.27 (d, J= 8 Hz, 2H), 7.14 (d, J= 8 Hz, 2H), 6.87 (br. s., 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.33-3.57 (m, 4H), 3.05 (s, 2H), 3.02 (s, 1H), 2.26 (s, 3H), 1.52-1.73 (m, 4H), 0.75-0.97 (m, 5H). MS
obsd. (ESE') [(M+H)+]: 460.
Example 37-A and Example 37-B
6-Amino-2-1S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one and 6-amino-2-1S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one 0 NDNH2 NI-12()ND
N'\........N
Nj\õ.....N
I-INN\ s, I )=0 cNIN oL 0 ,S ' N N S
0' 0 I-INV
.
Example 37-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A and pyrrolidine-l-carbonyl chloride instead of 6-amino-9-benzy1-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 37-A (390 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.31 -7.11 (m, 4H), 7.04 (s, 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.65 - 3.47 (m, 4H), 3.37 (m, 2H), 2.27 (s, 3H), 1.97 - 1.81 (m, 4H), 1.71 - 1.59 (m, 2H), 0.94 (t, J=
7.4 Hz, 3H). MS obsd. (ESE') [(M+H)+]: 458.2.
Example 37-B (125 mg) was prepared in analogy to Example 37-A by using Compound 36g-B instead of Compound 36g-A. 1H NMR (400 MHz, DMSO-d6) gppm:
7.28 - 7.14 (m, 4H), 7.04 (s, 2H), 4.95 (s, 2H), 4.15 (s, 1H), 3.65 - 3.47 (m, 4H), 3.37 (m, 2H), 2.27 (s, 3H), 1.93 - 1.84 (m, 4H), 1.65 - 1.60 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESE') [(M+H)+]: 458.3.
Example 38-A and Example 38-B
6-Amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2-1S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide and 6-amino-N-(2-methoxyethyl)-N-methyl-8-oxo-2-IS(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)purine-7-carboxamide N I-12L'N NH2L*N
N N
FINN\ s, 0 ,S 's N N S = N N
0' . HN
*
Example 38-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 36g-A and N-(2-methoxyethyl)-N-methyl-carbamoyl chloride (Intermediate AB) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy0-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA).
Example 38-A (57.8 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.26 (d, J= 7.6 Hz, 2H), 7.14 (d, J= 7.6 Hz, 2H), 6.89 - 6.78 (m, 2H), 4.95 (s, 2H), 4.18 (s, 1H), 3.62 -3.58 (m, 2H), 3.43 - 3.37 (m, 2H), 3.30 -3.10 (m, 3H), 3.09 - 3.08 (m, 3H), 3.08 - 3.05 (m, 2H), 2.27 (s, 3H), 1.77 - 1.54 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 476.3.
Example 38-B (46.6 mg) was prepared in analogy to Example 38-A by using Compound 36g-B instead of Compound 36g-A. 1H NMR (400 MHz, DMSO-d6) gppm:
7.26 (d, J= 7.6 Hz, 2H), 7.14 (d, J= 7.6 Hz, 2H), 6.89 - 6.78 (m, 2H), 4.95 (s, 2H), 4.18 (s, 1H), 3.62 -3.58 (m, 2H), 3.43 - 3.37 (m, 2H), 3.30 - 3.10 (m, 3H), 3.09 - 3.08 (m, 3H), 3.08 - 3.05 (m, 2H), 2.27 (s, 3H), 1.77 - 1.54 (m, 2H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESL') [(M+H)+]: 476.3.
Example 39 6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide 0 r----NH2 ....... N
\
N
NJ"
9µ I
//Sµµ N N
NH
it The title compound was prepared in analogy to Example 1, Method A, Step 6 by using N-ethyl-N-methyl-carbamoyl chloride and 6-amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36g) instead of N-methyl-N-propyl-carbamoyl chloride (Intermediate AA) and 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide (141.8 mg, Example 39) was obtained as a light yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.26 (d, J = 7.9 Hz, 2H), 7.15 (d, J =
7.9 Hz, 2H), 6.89 (s, 2H), 4.95 (s, 2H), 4.24 - 4.07 (m, 1H), 3.52 - 3.35 (m, 4H), 3.10 - 2.95 (m, 3H), 2.26 (s, 3H), 1.77 - 1.55 (m, 2H), 1.24 - 1.10 (m, 3H), 0.95 (t, J= 7.4 Hz, 3H). MS
obsd. (ESL') [(M+H)+]: 446.1.
Example 40 6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-carboxamide 0 rr \
N
N.--N
*
N
H
The title compound was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 36g) and N-butyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide (32 mg, Example 40) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.28 - 7.14 (m, 4H), 6.88 (s, 2H), 4.95 (s, 2H), 4.16 (s, 1H), 3.41 - 3.36 (m, 2H), 3.10 - 2.99 (m, 3H), 2.53 - 2.51 (m, 2H), 2.27 (s, 3H), 1.71 - 1.63 (m, 2H), 1.62 - 1.51 (m, 2H), 1.42 - 1.26 (m, 2H), 0.97 - 0.74 (m, 6H). MS obsd. (ESI ) [(M+H)+]: 474.3 Example 41-A and Example 41-B
6-Amino-9-[(4-chlorophenyl)methy1]-2-IS(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A) and 6-Amino-9-[(4-chlorophenyl)methy1]-2-IS(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B) ,.., NH2'-'N N H2 N
NNo NI---"'N
OµN oL j., H 1\lµN s, I >==o S ' N N ,S
I-INV 0' Step 1: Preparation of 6-amino-9-[(4-chlorophenyl)methyl]-2-ethylsulfany1-7H-purin-8-one (Compound 41a) H
NCN
I
S)N N
41a Compound 41a was prepared in analogy to Example 1, Method A, Step 3 by using iodoethane and 6-amino-9-[(4-chlorophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 34b) instead of bromopropane and 6-amino-9-phenylmethy1-2-sulfany1-purin-8-one (Compound lb). 6-Amino-9-[(4-chlorophenyl)methy1]-2-ethylsulfany1-purin-8-one (2.5 g, Compound 41a) was obtained as a white solid. MS obsd.
(ESL') [(M+H)+]: 336.
Step 2: Preparation of 6-amino-9-(4-chlorobenzy1)-2-ethylsulfmy1-7H-purin-8-one (Compound 41b) H
I /C) ii it CI
41b Compound 41b was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-9-[(4-chlorophenyl)methy1]-2-ethylsulfany1-7H-purin-8-one (Compound 41a) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-9-(4-chlorobenzy1)-2-ethylsulfiny1-7H-purin-8-one (1.94 g, Compound 41b) was obtained as a white solid. MS obsd. (ESI ) [(M+H)+]: 352.
Step 3: Preparation of 6-amino-9-[(4-chlorophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 41c) )Z
NN
SNN
NH
ilt CI
41c Compound 41c was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-9-(4-chlorobenzy1)-2-ethylsulfiny1-7H-purin-8-one (Compound 41b) instead of 6-amino-9-benzy1-2-(2-methylsulfiny1)-7H-purin-8-one (Compound 1d). 6-Amino-9-[(4-chlorophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (217 mg, Example 41c) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 10.61 (s, 1H), 7.42 -7.35 (m, 4H), 6.98 (s, 2H), 4.96 (s, 2H), 4.05 (s, 1H), 3.42 - 3.37 (m, 2H), 1.16 (t, J = 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 367Ø
Separation of compound of Compound 41c by chiral HPLC afforded Compound 41c-A (faster eluting, 31.8 mg) and Compound 41c-B (slower eluting, 10 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak IC-3 column.
N
FINN, ,S = N N
0' a 41c-A
Compound 41c-A: 1H NMR (400 MHz, DMSO-d6) gppm: 10.76 (s, 1H), 7.45 - 7.33 (m, 4H), 7.01 (s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.40 - 3.34 (m, 2H), 1.17 (t, J
= 7.4 Hz, 3H).
MS obsd. (ESL') [(M+H)+]: 367Ø
NN
)=0 ,S = N N
H
CI
41c-B
Compound 41c-B: 1H NMR (400 MHz, DMSO-d6) gppm: 10.70 (s, 1H), 7.46 -7.28 (m, 4H), 7.01 (s, 2H), 4.96 (s, 2H), 4.03 (s, 1H), 3.44 - 3.36 (m, 2H), 1.17 (t, J= 7.4 Hz, 3H).
MS obsd. (ESL') [(M+H)+]: 367Ø
Step 4: 6-Amino-9-[(4-chlorophenyl)methy1]-2-1S(R)-ethylsulfonimidoyll-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A) and 6-amino-9-[(4-chlorophenyl)methy1]-2-1S(S)-ethylsulfonimidoyll-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B) _ NI-12L*N N I-120 N
N N
N¨' N--' % ,I j., )=0 I-1 NN, ,, I
,s' s N N ,s' N N
I-11\V 0' * CI = CI
Example 41-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-9-[(4-chlorophenyOmethy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-A, 78 mg) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 5.00 (s, 2H), 4.19 (s, 1H), 3.46-3.39 (m, 2H), 3.39 - 3.38 (m, 2H), 3.09-2.99 (m, 3H), 1.69 -1.52 (m, 2H), 1.19 (t, J= 7.28 Hz, 3H), 0.95 - 0.66 (m, 3H). MS obsd. (ESL') [(M+H)+]:
466.1.
Example 41-B (125 mg) was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-purin-8-one (Compound 1 e ). 6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 41-B, 38 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 5.00 (s, 2H), 4.20 (s, 1H), 3.46 - 3.41 (m, 2H), 3.40 -3.39 (m, 2H), 3.10 - 3.00 (m, 3H), 1.69 - 1.50 (m, 2H), 1.24 - 1.12 (m, 3H), 0.93 - 0.73 (m, 3H).
(MS obsd.
(ESL') [(M+H)+]: 466.2.
The stereochemistry of Example 41-B was determined by single crystal X-ray diffraction shown in Figure 1.
Example 42-A and Example 42-B
6-Amino-9-[(4-chlorophenyl)methyll-N-ethy1-2[S(S)-ethylsulfonimidoyll-N-methy1-oxo-purine-7-carboxamide (Example 42-A) and 6-Amino-9-[(4-chlorophenyl)methyll-N-ethy1-2-1S(R)-ethylsulfonimidoyll-N-methy1-8-oxo-purine-7-carboxamide (Example 42-B) , r N H2Ly N
r2uyN
N'N
N
I-1 NINN ,, I )= N -----,s . N N
,_'s -N ¨NN 0' H NV )0 CI
. ci Example 42-A was prepared in analogy to Example 1, Method A, step 6 by using Compound 41c-A and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide (Example 42-A, 40 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.90 (s, 2H), 4.99 (s, 2H), 4.18 (s, 1H), 3.48 - 3.40 (m, 2H), 3.39 (s, 2H), 3.05 ¨3.01 (m, 3H), 1.20 - 1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 452.2.
Example 42-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 41c-B and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[5(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide (Example 42-B, 38 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) gppm: 7.43 - 7.41 (m, 4H), 6.91 (s, 2H), 4.98 (s, 2H), 4.19 (s, 1H), 3.48 - 3.40 (m, 2H), 3.39 (s, 2H), 3.09 -2.97 (m, 3H), 1.23- 1.11 (m, 6H). MS obsd. (ESE') [(M+H)+]: 452.2.
The stereochemistry of Example 42-A was determined by single crystal X-ray diffraction shown in Figure 2.
Example 43-A and Example 43-B
6-Amino-2-1S(R)-ethylsulfonimidoy11-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A) and 6-Amino-2-1S(S)-ethylsulfonimidoy11-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B) NJ---N
oNN s, 0 NJ'N
N N H N
NN s, 0 S ' * NV ,S . N N
HI
0' Step 1: Preparation of 4-amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (Compound 43a) H
N
N _ 3__Nru N
*H 2N
43a Compound 43a was prepared in analogy to Example 1, Method A, Step 1 by using 4-methylbenzyl isocyanate instead of benzyl isocyanate. 4-Amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (26.6 g, Compound 43a) was obtained as a grey solid and used directly for next step without further purification. MS obsd.
(ESL') [(M+H)+]: 229.2.
Step 2: Preparation of 6-amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (Compound 43b) H
N )j1:1 NI\_o H S N N
*
43b Compound 43b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-2-oxo-3-(p-tolylmethyl)-1H-imidazole-5-carbonitrile (compound 43a) instead of 4-amino-3-benzy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (20.0 g, Compound 43b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 288.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43c) H
N
.SIN N
#
43c Compound 43c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-(p-tolylmethyl)-2-sulfany1-7H-purin-8-one (Compound 43b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (13 g, Compound 43c) was obtained as a yellow solid. MS obsd. (ESE) [(M+H)+]: 316.
Step 4: Preparation of 6-amino-2-ethylsulfmy1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43d) NI-1\11 SLI
N N
II
*
43d Compound 43d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-2-ethylsulfany1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43c) instead of 6-amino-9-benzy1-2-methylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-2-ethylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one6 (3.5 g, Compound 43d) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 332.
Step 5: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43e) H
NN
SN N
õ
N H
#
43e Compound 43e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-2-ethylsulfiny1-9-(p-tolylmethyl)-7H-purin-8-one (Compound 43d) instead of 6-amino-9-benzy1-2-methylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(ethylsulfonimidoy1)-9-(p-tolylmethyl)-7H-purin-8-one (530 mg, Compound 43e) was obtained as a yellow solid. 1H NMR (400 MHz, DMSO-d6) gppm: 10.53 (s, 1H), 7.24 (d, J
= 8.03 Hz, 2H), 7.13 (d, J= 8.03 Hz, 2H), 6.94 (br. s., 2H), 4.91 (s, 2H), 4.03 (s, 1H), 3.36 -3.41 (m, 2H), 2.26 (s, 3H), 1.18 (t, J= 7.28 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 347.
Separation of compound of Compound 43e by chiral HPLC afforded Compound 43e-A (faster eluting, 56.8 mg) and Compound 43e-B (slower eluting, 56.7 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
H
N) --N
oNN 0 ,S
I-INV
=
43e-A
Compound 43e-A: 1H NMR (400 MHz, DMSO-d6) gppm: 10.52 (br. s., 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J = 7.9 Hz, 2H), 6.94 (br. s., 2H), 4.90 (s, 2H), 4.03 (s, 1H), 3.42 -3.33 (m, 2H), 2.25 (s, 3H), 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
347.
H
NN
FINN\ L I O
,S
0' y 43e-B
Compound 43e-B: 1H NMR (400 MHz, DMSO-d6) gppm: 10.56 (br. s., 1H), 7.23 (d, J=
8.0 Hz, 2H), 7.13 (d, J = 8.0 Hz, 2H), 6.95 (br. s., 2H), 4.90 (s, 2H) 4.03 (s, 1H), 3.44 -3.29 (m, 2H), 2.25 (s, 3H), 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]:
347.
Step 6: Preparation of 6-Amino-2-1S(R)-ethylsulfonimidoyli-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A) and 6-Amino-2-1S(S)-ethylsulfonimidoyli-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B) _ d NI H2L*N
NH2uN
N
N I-1 N N..
).-0 s, 0 N
\NNN I 0 ,S ' N N
HIN
..s ' N N ,, 0' S .
Example 43-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-A instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-A, 58.1 mg, faster eluting, isopropanol from 5% to 40% (0.05%DEA)/CO2 on ChiralPak AD-3 column) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.8 Hz, 2H), 7.15 (d, J=
7.8 Hz, 2H), 6.88 (br. s., 2H), 5.03 - 4.87 (m, 2H), 4.19 (s, 1H), 3.61 -3.36 (m, 4H), 3.11 ¨ 2.96 (m, 3H), 2.26 (s, 3H), 1.72 - 1.45 (m, 2H), 1.20 (t, J= 7.2 Hz, 3H), 0.97 - 0.65 (m, 3H). MS
obsd. (ESL') [(M+H)+]: 446.
Example 43-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-B instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide (Example 43-B, 40.1 mg, slower eluting, isopropanol from 5% to 40% (0.05%DEA)/CO2 on ChiralPak AD-3 column) was obtained as a white solid: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.5 Hz, 2H), 7.15 (d, J =
7.5 Hz, 2H), 6.89 (br. s., 2H), 5.03 - 4.86 (m, 2H), 4.19 (s, 1H), 3.49 - 3.37 (m, 4H), 3.08 - 3.00 (m, 3H), 2.27 (s, 3H), 1.70 - 1.48 (m, 2H), 1.20 (t, J= 7.2 Hz, 3H), 0.95 - 0.71 (m, 3H). MS
obsd. (ESE') [(M+H)+]: 446.3.
The stereochemistry of Example 43-B was determined by single crystal X-ray diffraction shown in Figure 3.
Example 44-A and Example 44-B
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoyli-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-A) and 6-Amino-N-ethy1-2-1S(R)-ethylsulfonimidoyli-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-B) r NNµ oL
,S N N SN N
Example 44-A was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-B and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2[5(5)-ethylsulfonimidoy1]-N-methy1-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-A, 73.1 mg) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J = 7.8 Hz, 2H), 7.15 (d, J= 7.8 Hz, 2H), 6.90 (br. s., 2H), 4.95 (s, 2H), 4.19 (br. s., 1H), 3.48 - 3.39 (m, 4H), 3.06 - 3.00 (m, 3H), 2.27 (s, 3H), 1.29 - 1.04 (m, 6H). MS obsd. (ESE') [(M+H)+]: 432.
Example 44-B was prepared in analogy to Example 1, Method A, Step 6 by using Compound 43e-A and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzyl-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide (Example 44-B, 46.7 mg) was obtained as a white solid: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.28 (d, J= 7.9 Hz, 2H), 7.15 (d, J= 7.9 Hz, 2H), 6.90 (br. s., 2H), 4.95 (s, 2H), 4.19 (br. s., 1H), 3.50 - 3.39 (m, 4H), 3.10 ¨ 2.96 (m, 3H), 2.27 (s, 3H), 1.27- 1.10 (m, 6H). MS obsd. (ESL') [(M+H)+]:
432.
Example 45-A and Example 45-B
6-Amino-2-IS (R) ethylsulfonimidoy1]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide and 6-Amino-2-1S(S)ethylsulfonimidoy1]-9-1(4-fluorophenyl)methyl]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide HNµN
N N ,S N N
H
F F
Step 1: Preparation of 4-amino-3-1(4-fluorophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 45a) H2N * F
45a Compound 45a was prepared in analogy to Example 1, Method A, Step 1 by using 4-fluorobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-fluorophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (48 g, Compound 45a) was obtained as a light yellow solid and was used directly for next step without further purification. MS obsd. (ESL') [(M+H)+]: 233.
Step 2: Preparation of 6-amino-9-[(4-fluorophenyl)methyl]-2-sulfany1-7H-purin-8-one (Compound 45b) H
NJ il I
H S C N N
* F
45b Compound 45b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-3-[(4-fluorophenyOmethy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 45a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-[(4-fluorophenyl)methy1]-2-sulfany1-7H-purin-8-one (32.0 g, Compound 45b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 292.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45c) N)j:N
SIN N
45c Compound 45c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-fluorophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 45b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (5.6 g, Compound 45c) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]:
320.
Step 5: Preparation of 6-amino-2-ethylsulfmy1-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45d) NJ11-1\11 S(I
N N
F
45d Compound 45d was prepared in analogy to Example 1, Method A, Step 4 by using 6-amino-2-ethylsulfany1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45c) instead of 6-amino-9-benzy1-2-propylsulfany1-7H-purin-8-one (Compound 1c). 6-Amino-2-ethylsulfiny1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (4.8 g, Compound 45d) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 332.
Step 6: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45e) N
I
H N
* F
45e Compound 45e was prepared in analogy to Example 1, Method A, Step 5 by using 6-amino-2-ethylsulfiny1-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45d) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (2.9 g, Compound 45e) was obtained as a yellow solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.57 (br.
s., 1H), 7.40 (dd, J= 8.5, 5.5 Hz, 2H), 7.16 (t, J= 8.9 Hz, 2H), 6.97 (br. s., 2H), 4.94 (s, 2H), 4.07 (s, 1H), 3.43 - 3.36 (m, 2H), 1.17 (t, J= 7.4 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 351.
Separation of compound of Compound 45e by chiral HPLC afforded Compound 45e-A (faster eluting, 85.4 mg) and Compound 45e-B (slower eluting, 36.4 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 45e-A: 1H NMR (400 MHz, DMSO-d6)6 ppm: 10.53 (br. s., 1H), 7.41 (dd, J=
8.5, 5.5 Hz, 2H), 7.17 (t, J= 8.9 Hz, 2H), 6.98 (br. s., 2H), 4.95 (s, 2H), 4.07 (s, 1H), 3.45-3.36 (m, 2H), 1.17 (t, J = 7.3 Hz, 3H). MS obsd. (ES[) [(M+H)+]: 351.
Compound 45e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.53 (br. s., 1H), 7.41 (dd, J=
8.5, 5.5 Hz, 2H), 7.17 (t, J= 8.9 Hz, 2H), 6.98 (br. s., 2H), 4.95 (s, 2H), 4.07 (s, 1H), 3.44 -3.37 (m, 2H) 1.17 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 351.
Step 7: Preparation of 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 45), 6-Amino-2-IS (R)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-Amino-2-1S(S)ethylsulfonimidoyl]-9-1(4-fluorophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 45-A and Example 45-B).
N I-12L* N
ON, I
N
I-IN' F
(Example 45) NI11-12`-*N
0N\ I I 1-INNµ I )=0 N N SN N
HN- 0' y F = F
(Example 45-A and Example 45-B) Example 45 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-7H-purin-8-one (Compound 45e) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide (162.4 mg, Example 45) was obtained as a white solid.
Separation of compound of Example 45 by chiral HPLC afforded Example 45-A
(faster eluting, 85.3 mg) and Example 45-B (slower eluting, 52 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 45-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.53 -7.38 (m, 2H), 7.18 (t, J= 8.9 Hz, 2H), 6.90 (br. s., 2H), 4.99 (s, 2H), 4.21 (s, 1H), 3.48 - 3.37 (m, 4H), 3.10 -3.01 (m, 3H), 1.69 - 1.49 (m, 2H), 1.25 - 1.14 (m, 3H), 0.94 - 0.72 (m, 3H).
MS obsd.
(ESL') [(M+H)+]: 450.
Example 45-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.54 - 7.38 (m, 2H), 7.18 (t, J= 8.9 Hz, 2H), 7.01 - 6.72 (m, 2H), 4.99 (s, 2H), 4.21 (s, 1H), 3.46 - 3.38 (m, 4H), 3.10 -3.01 (m, 3H), 1.76 - 1.50 (m, 2H), 1.25 - 1.16 (m, 3H), 0.99 - 0.69 (m, 3H).
MS obsd.
(ESI ) [(M+H)+]: 450.
Example 46-A and Example 46-B
6-Amino-N-ethyl-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methyl-8-oxo-purine-7-carboxamide (Example 46), 6-amino-N-ethyl-2-1S(S)-(ethylsulfonimidoy1)1-9-[(4-fluorophenyl)methy1]-N-methyl-8-oxo-purine-7-carboxamide and 6-amino-N-ethyl-2-1S(R)-(ethylsulfonimidoy1)1-9-[(4-fluorophenyl)methyl]-N-methyl-8-oxo-purine-7-carboxamide (Example 46-A and Example 46-B).
r NH20yN
N
N"
oNµ I 0 SN N
. F
(Example 46) r r NI-12L', N NH2 N
N.,....N N N
J----H NNµ L 1 oNN s,I I 0 ,S ' N N S ' N N
0' y H NV
. F / . F
(Example 46-A and Example 46-B) Example 46 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-7H-purin-8-one (Compound 45e) and N-ethyl-N-methyl carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-purine-7-carboxamide (51 mg, Example 46) was obtained as a white solid. 'H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.15 (m, 2H), 6.90 (br. s., 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.47 - 3.32 (m, 4H), 3.05 -3.01 (m, 3H), 1.21 - 1.14 (m, 6H). MS obsd. (ESI ) [(M+H)+]: 436.
Separation of compound of Example 46 by chiral HPLC afforded Example 46-A
(faster eluting, 72 mg) and Example 46-B (slower eluting, 45 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 46-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.16 (m, 2H), 6.90 (br. s., 2H), 4.98 (s, 2H), 4.18 (s, 1H), 3.47 - 3.32 (m, 4H), 3.05 - 3.01 (m, 3H), 1.21-1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 436.
Example 46-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.46 - 7.43 (m, 2H), 7.20-7.14 (m, 2H), 6.92 (br. s., 2H), 4.98 (s, 2H), 4.20 (br. s., 1H), 3.47 - 3.32 (m, 4H), 3.05 -3.01 (m, 3H), 1.23 - 1.19 (m, 6H). MS obsd. (ESL') [(M+H)+]: 436.
Example 47-A and Example 47-B
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47), 6-amino-2-IS(R)-ethylsulfonimidoy1]-[(4-bromophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-amino-2-IS(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methyll-N-methyl-8-oxo-N-propyl-purine-7-carboxamide NFIPN
N------N
,sN N
HN' . Br (Example 47) N1-12 N NHPyN
N--N N)--' N
% s,I )=0 1-1N (:) S
,,L I
S ' N N s' NN
HNV CV
0 Br . Br (Example 47-A and Example 47-B) Step 1: Preparation of 4-amino-3-[(4-bromophenyl)methyl]-2-oxo-1H-imidazole-5-carbonitrile (Compound 47a) N H
. Br 47a Compound 47a was prepared in analogy to Example 1, Method A, Step 1 by using 4-bromobenzyl isocyanate instead of benzyl isocyanate. 4-Amino-3-[(4-bromophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (500 mg, Compound 47a) was obtained as a light yellow solid and was used directly for next step without further purification. 1H NMR (400 MHz, DMSO-d6) 5 ppm: 9.94 (S, 1H), 7.55-7.53 (d, J=
8.0 Hz, 2H), 7.20-7.18 (d, J = 8.0 Hz, 2H), 6.52 (br. s., 2H), 4.74 (s, 2H). MS obsd.
(ESI ) [(M+H)+]: 293.
Step 2: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 47b) N- N
1 )=0 HS NN
. Br 47b Compound 47b was prepared in analogy to Example 1, Method A, Step 2 by using of 4-amino-3-[(4-bromophenyl)methy1]-2-oxo-1H-imidazole-5-carbonitrile (Compound 47a) instead of 4-amino-3-phenylmethy1-2-oxo-1H-imidazole-5-carbonitrile (Compound la). 6-Amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (300 mg, Compound 47b) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]: 352.
Step 3: Preparation of 6-amino-2-ethylsulfany1-9-[(4-bromophenyl)methyl]-7H-purin-8-one (Compound 47c) Ni Nso S NN
) *Br 47c Compound 47c was prepared in analogy to Example 1, Method A, Step 3 by using 6-amino-9-[(4-bromophenyl)methy1]-2-sulfany1-7H-purin-8-one (Compound 45b) and iodoethane instead of 6-amino-9-benzy1-2-sulfany1-7H-purin-8-one (Compound lb) and bromopropane. 6-Amino-2-ethylsulfany1-9-[(4-bromophenyOmethyl]-7H-purin-8-one (5.6 g, Compound 47c) was obtained as a yellow solid. MS obsd. (ESL') [(M+H)+]:
380.
Step 4: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfmyl-7h-purin-8-one (compound 47d) N:N
S)N N
ii * Br 47d Compound 47d was prepared in analogy to Example 1, Method B, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfany1-7H-purin-8-one ( Compound 47c) instead of 6-amino-9-benzy1-2-(2-propylsulfany1)-7H-purin-8-one (Compound 1c).
Amino-9-[(4-bromophenyl)methy1]-2-ethylsulfiny1-7H-purin-8-one (3.2 g, Compound 47d) was obtained as a white solid. MS obsd. (ESL') [(M+H)+]: 396.
Step 5: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7h-purin-8-one (compound 47e) H
N N
0 1 >0 \ \ ;.........
H N= S N N
) . Br 47e Compound 47e was prepared in analogy to Example 1, Method B, Step 7 by using 6-amino-9-[(4-bromophenyl)methy1]-2-ethylsulfiny1-7H-purin-8-one (Compound 47d) instead of 6-amino-9-benzy1-2-propylsulfiny1-7H-purin-8-one (Compound 1d). 6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (4.0 g, Compound 47e) was obtained as a white solid. MS obsd. (ESE') [(M+H)+]: 411.
N NH N
ssL 1-1No I
S N S N N
H N 0' = Br 1>
Br Compound 47e-A and Compound 47e-B
Separation of compound of Compound 47e by chiral HPLC afforded Compound 47e-A (faster eluting, 112 mg) and Compound 47e-B (slower eluting, 99 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Compound 47e-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.58 (br. s., 1H), 7.52-7.54 (d, J= 8.0, 2H), 7.31-7.29 (t, J= 8.0 Hz, 2H), 6.54 (br. s., 2H), 4.93 (s, 2H), 4.05 (s, 1H), 3.42 -3.31 (m, 2H), 1.15 (t, J= 7.3 Hz, 3H). MS obsd. (ESI ) [(M+H)+]: 411.
Compound 47e-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 10.58 (br. s., 1H), 7.54-7.52 (d, J= 8.0, 2H), 7.31-7.29 (t, J= 8.0 Hz, 2H), 6.98 (br. s., 2H), 4.93 (s, 2H), 4.06 (s, 1H), 3.40 -3.37 (m, 2H), 1.15 (t, J= 7.3 Hz, 3H). MS obsd. (ESL') [(M+H)+]: 411.
Step 6: Preparation of 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47), 6-amino-9-[(4-bromophenyl)methy1]-2-1S(R)-ethylsulfonimidoyli-N-methyl-8-oxo-N-propyl-purine-7-carboxamide and 6-amino-9-[(4-bromophenyl)methy1]-2-1S(S)-ethylsulfonimidoyli-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (Example 47-A and Example 47-B).
N I-12L*
NN
I-11\V
= Br (Example 47) NH2uy N"
oL HNx, N N
Br Br (Example 47-A and Example 47-B) Example 47 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 47e) instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le). 6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (570 mg, Example 47) was obtained as a white solid. 1H
NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.53 (m, 2H), 7.36-7.34 (m, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.18 (s, 1H), 3.45 - 3.38 (m, 4H), 3.05 - 3.02 (m, 3H), 1.65-1.56 (m, 2H), 1.19 (t, J= 8.0 Hz, 3H), 0.93-0.75 (m, 3H). MS obsd. (ESL') [(M+H)+]: 510.
Separation of compound of Example 47 by chiral HPLC afforded Example 47-A
(faster eluting, 260 mg) and Example 47-B (slower eluting, 266 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column Example 47-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.33 (d, J= 8,0 Hz, 2H), 6.90 (br. s., 2H), 4.97 (s, 2H), 4.21 (s, 1H), 3.46 - 3.41 (m, 4H), 3.05 - 3.02 (m, 3H),1.65-1.54 (m, 2H), 1.24-1.16 (m, 3H), 0.93-0.75 (m, 3H). MS
obsd. (ESE') [(M+H)+]: 510.
Example 47-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.54 - 7.53 (d, J= 8.0 Hz, 2H), 7.36-7.33 (d, J= 8,0 Hz, 2H), 6.90 (br. s., 2H), 4.97 (s, 2H), 4.21 (s, 1H), 3.46 - 3.41 (m, 4H), 3.06 - 3.02 (m, 3H), 1.65-1.54 (m, 2H), 1.20-1.16 (m, 3H), 0.93-0.75 (m, 3H).
MS obsd. (ESI ) [(M+H)+]: 510.
Example 48-A and Example 48-B
6-Amino-9-[(4-bromophenyl)methy1]-N-ethyl-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide (Example 48), 6-amino-9-[(4-bromophenyl)methy1]-N-ethyl-2-IS(S)-(ethylsulfonimidoy1)1-N-methyl-8-oxo-purine-7-carboxamide and 6-amino-9-[(4-bromophenyl)methyll-N-ethy1-2-1S(R)-(ethylsulfonimidoy1)]-N-methyl-8-oxo-purine-7-carboxamide (Example 48-A and Example 48-B).
NH2 o.....r T \
N-1\1 SN----N
II
= Br (Example 48) 0 r---NH2 .......N N H2 o.....r \ \
NJ-----N N----"N
N H4 (:) 0 1 >=0 0,1, , H Nk I I
S 's N-----N
= Br ic = Br (Example 48-A and Example 48-B) r r NH2 yN
NH2 ,,N
N)-----N 111\1µµ .0L I )=C) NN
-""k 0 0' S N N
H NV
ilt CI
Example 48 was prepared in analogy to Example 1, Method A, Step 6 by using 6-amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-7H-purin-8-one (Compound 47e) and N-ethyl-N-methyl-carbamoyl chloride instead of 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7H-purin-8-one (Compound le) and N-methyl-N-propyl-carbamoyl chloride (Intermediate AA). 6-Amino-9-[(4-bromophenyOmethyl]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide (469 mg, Example 48) was obtained as a white solid. 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 -7.54 (d, J
= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.98 (br. s., 2H), 4.97 (s, 2H), 3.53 - 3.46 (m, 4H), 3.05 - 3.01 (m, 3H), 1.22-1.16 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Separation of compound of Example 48 by chiral HPLC afforded Example 48-A
(faster eluting, 198 mg) and Example 48-B (slower eluting, 202 mg) as white solid with methanol 5%-40% (0.05%DEA)/CO2 on ChiralPak AD-3 column.
Example 48-A: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.19 -4.18 (m, 1H), 3.46 - 3.41 (m, 4H), 3.05 - 3.01 (m, 3H), 1.20-1.14 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Example 48-B: 1H NMR (400 MHz, DMSO-d6) 6 ppm: 7.56 - 7.54 (d, J= 8.0 Hz, 2H), 7.36-7.34 (d, J= 8,0 Hz, 2H), 6.92 (br. s., 2H), 4.97 (s, 2H), 4.24 (br.
s., 1H), 3.58 -3.41 (m, 4H), 3.05 - 3.01 (m, 3H), 1.26-1.01 (m, 6H). MS obsd. (ESL') [(M+H)+]: 496.
Example 49 Activity of Compounds and Examples in HEK293-hTLR-7 assay HEK293-Blue-hTLR-7 cells assay:
A stable HEK293-Blue-hTLR-7 cell line was purchased from InvivoGen (Cat.#:
hkb-ht1r7, San Diego, California, USA). These cells were designed for studying the stimulation of human TLR7 by monitoring the activation of NF-KB. A SEAP
(secreted embryonic alkaline phosphatase) reporter gene was placed under the control of the IFN-13 minimal promoter fused to five NF-KB and AP-1-binding sites. The SEAP was induced by activating NF-KB and AP-1 via stimulating HEK-Blue hTLR7 cells with TLR7 ligands.
Therefore the reporter expression was regulated by the NF-KB promoter upon stimulation of human TLR7 for 20 hrs. The cell culture supernatant SEAP reporter activity was determined using QUANTI-BlueTm kit (Cat.#: rep-qbl, Invivogen, San Diego, Ca, USA) at a wavelength of 640 rim, a detection medium that turns purple or blue in the presence of alkaline phosphatase.
HEK293-Blue-hTLR7 cells were incubated at a density of 250,000-450,000 cells/mL in a volume of 180 [EL in a 96-well plate in Dulbecco's Modified Eagle's medium (DMEM) containing 4.5 g/L glucose, 50 U/mL penicillin, 50 mg/mL streptomycin, mg/mL Normocin, 2 mM L-glutamine, 10% (V/V) heat-inactivated fetal bovine serum for 24 hrs. Then the HEK293-Blue-hTLR-7 cells were incubated with addition of 20 [EL test compound in a serial dilution in the presence of final DMSO at 1% and perform incubation under 37 C in a CO2 incubator for 20 hrs. Then 20 [EL of the supernatant from each well was incubated with 180 [EL Quanti-blue substrate solution at 37 C for 2 hrs and the absorbance was read at 620-655 nm using a spectrophotometer. The signalling pathway that TLR7 activation leads to downstream NF-KB activation has been widely accepted, and therefore similar reporter assay was also widely used for evaluating TLR7 agonist (Tsuneyasu Kaisho and Takashi Tanaka, Trends in Immunology, Volume 29, Issue 7, July 2008, Pages 329.sci; Hiroaki Hemmi et al, Nature Immunology 3, 196 - 200 (2002)).
The Compounds and Examples of the present invention were tested in HEK293-hTLR-7 assay for their TLR7 agonism activity as described herein and results are listed in Table 1. The Examples of prodrugs were found to have EC50 of about 2.1 [EM to about 1000 [EM, the Compounds of active forms were found to have EC50 less than 0.2 [M. The calculated ratio of ECso(prodnio / ECso(active form) were within the range from 32 to about 7600.
Table 1. Activity of Examples and Compounds of present invention in HEK293-hTLR-7 assay Ratio hTLR-7 EC50 hTLR-7 EC50 Corresponding Prodrug (EC50(prodrug) /
Active Form (Prodrug, (Active form, EC50(active form)) ILLM) tiM) Example 1 50.4 Compound le 0.065 775.4 Example 1-A 42.5 Compound le-A 0.067 634.3 Example 1-B 27 Compound le-B 0.086 314.0 Example 2 32 Compound le 0.065 372.1 Example 2-A 3.7 Compound le-B 0.086 43.0 Example 2-B 4.4 Compound 1 e-A 0.067 65.7 Example 3 15.1 Compound le 0.065 232.3 Example 4 23 Compound le 0.065 353.8 Example 5 41 Compound le 0.065 630.8 Example 6 82.3 Compound le 0.065 1266.2 Example 7 19.9 Compound le 0.065 306.2 Example 8 2.1 Compound le 0.065 32.3 Example 9 19.2 Compound le 0.065 295.4 Example 10 68.5 Compound le 0.065 1053.8 Example 11 5.6 Compound le 0.065 86.2 Example 12 43.9 Compound le 0.065 675.4 Example 13 67 Compound le 0.065 1030.8 Example 14 2.4 Compound le 0.065 36.9 Example 15 494 Compound le 0.065 7600 Example 16 32.1 Compound le 0.065 493.8 Example 25 24.2 Compound le 0.065 372.3 Example 26 13.4 Compound le 0.065 206.2 Example 27 31.7 Compound le 0.065 487.7 Example 28 6.9 Compound le 0.065 106.2 Example 29 48.8 Compound le 0.065 750.8 Example 32 22.5 Compound le 0.065 346.2 Example 34- 6.0 Compound 34e-A 0.014 428.6 A
Example 34-6.36 Compound 34e-B 0.011 578.2 B
Example 36-A 31.8 Compound 36g-A 0.019 1673.7 Example 37-A 26.6 Compound 36g-A 0.019 1400 Example 37-B 47.4 Compound 36g-B 0.022 2154.5 Example 38-A 26.2 Compound 36g-A 0.019 1378.9 Example 38-B 19.5 Compound 36g-B 0.022 886.4 Example 39 4.3 Compound 36g 0.027 159.3 Example 40 52.8 Compound 36g 0.027 1955.6 Example 41 36 Compound 41c 0.053 679.2 Example 41-44.1 Compound 41c-B 0.085 518.8 A
Example 41-32.1 Compound 41c-A 0.071 452.1 B
Example 42-40.5 Compound 41c-A 0.071 570.4 A
Example 42-49.2 Compound 41c-B 0.085 578.8 B
Example 43-110 Compound 43e-A 0.11 1000 A
Example 43- 78.4 Compound 43e-B 0.035 2240 B
Example 44-65.4 Compound 43e-B 0.035 1868.6 A
Example 44-96.7 Compound 43e-A 0.11 879.1 B
Compound 45e-B
Example 45-153 Or 0.26 or 0.39 588 or 392 A
Compound 45e-A
Compound 45e-B
Example 45->1000 Or 0.26 or 0.39 >3846 or >2564 B
Compound 45e-A
Compound 45e-A
Example 46-45.5 Or 0.26 or 0.39 175 or 116.7 A
Compound 45e-B
Compound 45e-B
Example 46-45.7 Or 0.26 or 0.39 175.7 or 117.2 B
Compound 45e-A
Compound 47e-A
Example 47-10.9 Or 0.021 or 0.025 519.0 or A
Compound 47e-B
Compound 47e-A
Or Example 47-13.1 0.021 or 0.025 623.8 or B Compound 47e-B
Compound 47e-A
Example 48-18.3 Or 0.021 or 0.025 871.4 or A
Compound 47e-B
Compound 47e-A
Or Example 48-20.8 0.021 or 0.025 990.5 or B Compound 47e-B
Example 50 Metabolism of prodrugs of compound of formula (I) A study was undertaken to evaluate the metabolic conversion of prodrugs, compound of formula (I), to its corresponding active form. The compounds of formula (I), if served as prodrugs, can be metabolized to the active compound or other compounds of the invention in the body. Human liver microsomes are often used to assess the degree of metabolic conversion of prodrugs in the body of animal or human.
Materials NADPH cofactor system including 13-Nicotinamide adenine dinucleotide phosphate (NADP), isocitric acid and isocitric dehydrogenase were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). Human liver microsomes (Cat No. 452117, Lot No.
38290) were obtained from Corning (Woburn, MA, USA). Mouse liver microsomes (Cat No.
M1000, Lot No.1310028) were obtained from Xenotech.
Working solution of the compounds and other solution Compounds were dissolved in DMSO to make 10 mM stock solutions. 10 [EL of the stock solution was diluted with acetonitrile (990 [EL) to get a 100 [EM
working solution.
Incubation Microsomes were preincubated with test compound for 10 min at 37 C in 100 mM
potassium phosphate buffer with pH 7.4. The reactions were initiated by adding NADPH
regenerating system to give a final incubation volume of 200 [EL and shaken in a water bath at 37 C. Incubation mixtures consisted of liver microsomes (0.5 mg microsomal protein/mL), substrates (1.0 [tM), and NADP (1 mM), isocitric dehydrogenase(1 unit/mL), isocitric acid (6 mM).
Preparation of Samples for Analysis At 30 min, reaction was quenched by adding 600 [LL cold acetonitrile (including 100 ng/mL tolbutamide and 100 ng/mL labetalol as internal standard). The samples were centrifuged at 4000 rpm for 20 minutes and the resultant supernatants were subjected to LC-MS/MS analysis.
The samples for calibration curve were prepared as followed. Dispense 100 [LL/well liver microsomes and 98 [LL/well NADPH regenerating system solution to 96-well plate.
Add 600 [LL quenching solution first, and then followed by 2 [EL Standard curve and QC
working solution.
Bioanalysis The compounds were quantified on an API4000 LC-MC/MC instrument in the ESI-Positive MRM mode.
A study was undertaken to evaluate the metabolic conversion of prodrugs (1[EM), Example 1, Example 1-A, Example 1-B, Example 2, Example 2-A, Example 2-B, Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, Example 9, Example 10, Example 11, Example 12, Example 13, Example 14, Example 15, Example 16, Example 17, Example 21, Example 22, Example 23, Example 25, Example 26, Example 27, Example 28Example 29, Example 30, Example 31, Example 32, Example 33, Example 34-A, Example 34-B, Example 36-A, Example 36-B, Example 37-A, Example 37-B, Example 38-A, Example 38-B, Example 39, Example 40, Example 41, Example 41-A, Example 41-B, Example 42, Example 42-A, Example 42-B, Example 43, Example 43-A, Example 43-B, Example 44, Example 44-A, Example 44-B and Example 45-A, Example 46-A, Example 46-B, Example 47-A, Example 47-B, Example 48-A, Example 48-B to the corresponding active forms, Compound le, Compound le-A, Compound le-B, Compound 34e-A, Compound 34e-B, Compound 36g-A, Compound 36g-B, Compound 36g, Compound 41c, Compound 41c-B, Compound 41c-A, Compound 43e, Compound 43e-A, Compound 43e-B, Compound 45e-A, Compound 45e-B, Compound 47e-A, and Compound 47e-B in the presence of human liver microsomes. Results were summarized and shown in Table 2.
Table 2. Metabolic conversion of prodrugs in human liver microsomes Metaboli Metaboli zed zed product product Corresponding Corresponding concentr concentr Example Metabolized Metabolized ation in Example No.
ation in No. Product (active Product (active human human form) form) liver liver microso microso mes (?LM) mes (?LM) Example 1 Compound le 0.0214 Example 31 Compound le 0.005 Example 1- Compound le-A 0.018 Example 32 Compound le 0.013 Example 1- Compound le-B 0.022 Example 33 Compound le 0.59 Example 2 Compound le 0.028 Example 34-A Compound 34e-A 0.2 Example 2- Compound le-B 0.036 Example 34-B Compound 34e-B 0.088 Example 2- Compound le-A 0.029 Example 36-A Compound 36g-A 0.02 Example 3 Compound le 0.12 Example 36-B Compound 36g-B 0.019 Example 5 Compound le 0.078 Example 37-A Compound 36g-A 0.004 Example 6 Compound le 0.074 Example 37-B Compound 36g-B 0.002 Example 7 Compound le 0.15 Example 38-A Compound 36g-A 0.026 Example 8 Compound le 0.043 Example 38-B Compound 36g-B 0.034 Example 9 Compound le 0.002 Example 40 Compound 36g 0.032 Example 10 Compound le 0.005 Example 41-A Compound 41c-B 0.38 Example 11 Compound le 0.001 Example 41-B Compound 41c-A 0.36 Example 12 Compound le 0.018 Example 42-A Compound 41c-A 0.14 Example 13 Compound le 0.04 Example 42-B Compound 41c-B 0.004 Example 14 Compound le 0.026 Example 43-A Compound 43e-A 0.014 Example 15 Compound le 0.002 Example 43-B Compound 43e-B 0.016 Example 16 Compound le 0.024 Example 44-A Compound 43e-B 0.002 Example 17 Compound le 0.075 Example 44-B Compound 43e-A 0.002 Example 21 Compound le 0.48 Example 45-A Compound 45e-B 0.41 Or Example 22 Compound le 0.42 Example 46-A Compound 45e-A 0.039 Or Example 23 Compound le 0.42 Example 46-B Compound 45e-B 0.18 Or Example 25 Compound le 0.018 Example 47-A Compound 47e-A 0.36 Or Example 26 Compound le 0.042 Example 47-B Compound 47e-B
0.41 Or Example 27 Compound le 0.11 Example 48-A Compound 47e-A 0.11 Or Example 28 Compound le 0.084 Example 48-B Compound 47e-B
0.053 Or Example 29 Compound le 0.009 Example 51 In vivo combined efficacy (tumor free mice) of an prodrug form of compounds of the present invention (Compound 41-A) and sorafenib in a highly aggressive model of hepatocellular carcinoma In iAST mice tumorigenesis was initiated by intravenous injection of 5x108 IFU
adenovirus expressing Cre recombinase (Ad-CMV-iCre vector in vivo application, Vector Biolabs) into transgenic mice expressing the hepatocyte-specific albumin promoter, a loxP-flanked stop cassette, and the SV40 large T-antigen (Runge A, at al., Cancer Res. 74 (2014) 4157-69). The Cre recombinase excises the stop cassette in transduced cells and leads to a transient viral hepatitis and resulting in multinodular tumorigenesis within 8 weeks.
Female mice were treated with either vehicle (7.5% Gelatine / 0.22% NaCl for Sorafenib;
or 2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% Methylparaben (Sigma), 0.01%
Propylparabens (Sigma) in water for 41-A), or 90mg/kg in Sorafenib (Nexavar 0, Bayer HealthCare) daily or were treated with compound 41-A (10mg/kg) once a week by oral gavage. Treatment using vehicle or Sorafenib started on week 7.5 upon adenovirus administration and 3 days prior to administration compound 41-A. Animals were sacrificed on day 12 after treatment start and total liver and tumor weights were determined. Per group n=10 were analyzed by One-way ANOVA and Tukey correction shown as individual dots with means SEM using GraphPad Prism software version 6.
Although Sorafenib was highly effective in monotherapy, the combination with an active form of the compounds of the present invention (compound 41-A) resulted even in 2/10 tumor-free mice by superficial examination of the livers in this highly aggressive model of hepatocellular carcinoma. Results are shown in the Table below and in Figures lA and 1B
Synergistic effect of Compound 41-A and sorafenib on tumor burden ( tumor free mice) Treatment Examination of the liver for superficial tumor nodules Vehicle 0/10 tumor nodule free Compound 41-A 0/10 tumor nodule free Sorafenib 0/10 tumor nodule free Compound 41-A + 2/10 tumor nodule free Sorafenib Example 52 Treatment with an prodrug form of the compounds of the present invention (compound 41-A) induces PD-Li expression on tumor cells in hepatocellular carcinoma.
Tumors from iAST mice were treated as described in Figure 1. Animals were sacrificed on day 12 after treatment start and tumors analyzed by flow cytometry. For flow cytometry, tumors were excised and single cell suspensions obtained by mechanical processing and enzymatic digestion (DNAse 0.01%, Collagenase IV lmg/m1). Staining procedures started with Fc receptor blocking using 2.4G2 antibody clone (1:200 dilution, BD
Bioscience), and the following antibodies (clones) were used to analyzed leukocyte infiltrate: CD45-FITC (30-F11, BioLegend) and CD1 lb-BUV737 (M1/70, BD Bioscience). Samples were acquired using a LSR Fortessa machine (BD Bioscience) and analyzed by FlowJo version 10 (Treestar). Data are shown of n=5 per group, analyzed by One-way ANOVA and Tukey correction shown as individual dots with means SEM using GraphPad Prism software version 6. Although the absolute immune cell infiltrate in iAST tumor did not change by any treatments described (Fig.2A), significant changes were observed in the overall lymphoid and myeloid composition of the tumors (Fig. 2 C and D). Here, the changes were clearly driven by Sorafenib, which was previously shown to act also on immune cells (Martin del Campo, et al, J Immunol. 195 (2015) 1995-2005). 41-A treatment however induced PD-Li expression on tumor cells in monotherapy as well as in combination with Sorafenib (Fig. 2 B).
Example 53 In vivo triple combination of 41-A, Sorafenib and anti-PD-1 results in increased median survival.
Multinodular tumors were induced in iAST mice as described for Fig. 1 ( see Example 51).
Female transgenic mice were treated at 7.5 weeks upon virus injection with either vehicle (7.5% Gelatine / 0.22% NaCl for Sorafenib; or 2% Kluce10 Hydroxypropylcellulose LF
(Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% Methylparaben (Sigma), 0.01% Propylparabens (Sigma) in water for 41-A), or 90mg/kg in Sorafenib (Nexavar 0, Bayer HealthCare) daily or were treated with compound 41-A (10mg/kg) once a week by oral gavage. Treatment using vehicle or Sorafenib started on week 7.5 upon adenovirus administration and 3 days prior to administration compound 41-A. Anti-mouse PD-1 antibody (clone RPM1-14, BioXCell) was administered every 3 days intra peritoneally at 250[Eg/mouse. Total treatment period was 2 weeks (and 3 days + 2 weeks for Sorafenib) and survival of iAST mice was monitored. Mice were sacrificed upon display of distress signs such as >20%
gain of body weight, ruffled fur and or hatched position. Kaplan-Meier curves were analyzed by Pairwise Log-Rank test (see table). In the survival setting, neither Sorafenib, nor 41-A
were effective in monotherapy. Anti-PD-1 monotherapy even resulted in significantly reduced survival compared to VEH control. The median survival of iAST mice was significantly enhanced in the combination group of Sorafenib and anti-PD-1 antibody.
However, the triple combination of 41-A together with Sorafenib and anti-PD-1 led to the biggest and significant increase in median survival from 71 days (VEH) to 104 days (41-A
+ PD-1 + Sorafenib) in this highly aggressive HCC model. Results are shown in Figure 3 and the Table below Table: Pairwise Log-Rank Test (multiple test level = 0.00179) A +
+
Group VEH Sorafenib 41-A PD-1 41-A+ + PD- PD-1 PD-1 +
Sorafenib Sorafenib Sorafenib VEH 1.0 0.1580 0.2309 0.0004* 0.8759 0.1465 0.0192* 0.0001*
Example 54 Treatment with an prodrug form of the compounds of the present invention (compound 41-A) in the transplanted Hep55.1c mouse model of hepatocellular carcinoma Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20itl (10itl cell suspension plus 10itl Martigel).
Tumor volume was monitored weekly using itCT (TomoScope Synergy Twin, CT
Imaging GmbH) upon a single intravenous administration of contrasting agent Exitron (Viscovert). Imaging data were reconstructed by TomoScope software and analyzed using Osirix software. Once tumors reached 80mm3, mice were treated weekly with either 10mg/kg 41-A compound or vehicle (2% Kluce10 Hydroxypropylcellulose LF
(Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09%
Methylparaben (Sigma), 0.01% Propylparabens (Sigma) in water) per oral gavage.
To compare to another agonistic, immune stimulating agent, a single dose of anti-antibody (4mg/kg; clone FGK.45, BioXCell) was given. Data depicted are means SEM
for a minimum of n=9 animals per group.
Weekly administration of compound 41-A resulted in inhibition of tumor growth in Hep55.1c tumor bearing mice when compared to vehicle treatment. As previously published, a single dose of anti-CD40 antibody can lead to tumor eradication in subcutaneous MC38 tumors and it has been shown that the anti-CD40 antibody has an inflammatory effect in the liver (Hoves S, et al, J Exp Med, DOI:
10.1084/jem.20171440;
Published February 7, 2018). However, no beneficial treatment effect was observed in Hep55.1c tumor bearing mice with anti-CD40 antibody. Results are shown in Figure 5 A.
Example 55 In vivo efficacy of compound 42-A (6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide), alone and in combination with anti-PD-1 results in survival benefit in the Hep55.1c mouse model of hepatocellular carcinoma.
Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20 1 (10 1 cell suspension plus 10 1Martigel).
Scout animals were sacrificed to determine the time point of treatment start at about 80mm3 tumor volume. Mice were treated with either compound 42-A 10mg/kg or vehicle (2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol 1000 succinate (TPGS, Sigma), 0.09% methylparaben (Sigma), 0.01%
propylparaben (Sigma) in water) per oral gavage, or intra peritoneal administration of 250[tg anti-PD-1 antibody (clone RPM1-14, BioXCell), or a combination of compound 42-A plus anti-PD-1. 42-A was given weekly (total 3 times) and stated on the same day anti-PD-1 antibody treatment. Antibody treatment was continued every three to four days for 6 doses in total. Monotherapy with 42-A resulted smaller tumor volume compared to VEH
control and PD-1 monotherapy. Combined treatment of 42-A and anti-PD-1, tumor volume was also reduced with 3 out of 9 mice being tumor-free. Results are shown in Figure 5 B
and the Table below.
Combination effect of Compound 42-A and anti-PD-1 on tumor burden (tumor free mice) Treatment Examination of the liver Vehicle 0/7 tumor free aPD-1 1/9 tumor free 42-A 1/7 tumor free 42-A+aPD-1 3/9 tumor free Example 56 Combination of an prodrug form of the compounds of the present invention (compound 41-A) and anti-PD-1 antibodies Hep55.1c mouse model of hepatocellular carcinoma.
Female C57BL/6N mice (Jackson Laboratories) were injected intra hepatically with 5x105 Hep55.1c tumor cell line together with Matrigel (Matrigel Basement Membrane Matrix, Corning Cat# 354234) in a total volume of 20u1 (10u1 cell suspension plus 10 1 Martigel).
After 3 weeks, animals were sacrificed and tumors excised from the liver.
Excised tumors were cut into lx1 mm3 pieces and implanted into the liver of female C57BL/6N
mice.
Scout animals were sacrificed to determine the time point of treatment start at about 80mm3 tumor volume. Mice were treated with either 41-A or vehicle (2% Kluce10 Hydroxypropylcellulose LF (Asland), 0.5% D-a-Tocopherol polyethylene glycol succinate (TPGS, Sigma), 0.09% methylparaben (Sigma), 0.01% propylparaben (Sigma) in water) per oral gavage, or intra peritoneal administration of 250ug anti-PD-1 antibody (clone RPM1-14, BioXCell), or a combination of 41-1 plus anti-PD-1. 41-A was given weekly, while anti-PD-1 antibody treatment started one day after 41-A
treatment and was continued every three to four days for 8 doses in total. Treatment with both agents was stopped after the last anti-PD-1 administration. Monotherapy with 41-A
resulted in longer survival of mice (5/10) compared to vehicle control (1/10). Combined treatment of 41-A
and anti-PD-1 enhanced survival of mice even significantly to 8/10 being alive on day 94 after tumor fragment transplantation.
Example 57 Treatment with an active form of the compounds of the present invention (compound 41c-B) does not induce enhanced tumor cell proliferation in cell lines originating from hepatocellular carcinoma and cholangiocarcinoma Cell lines derived from hepatocellular carcinoma and cholangiocarcinoma (EGI1 and OZ) were maintained and tested in the following media: Huh7 and EGI1 were cultured in DMEM 4,5 g/L glucose (Gibco, Cat# 31966-021), 10 % FCS (GIBCO, Cat# 10500-064 Lot 07G3690K), 2 mM L-glutamine (Thermo Fischer, Cat# 25030081), 1mM sodium pyruvate (GIBCO Cat# 11360-039). Hep3B and HepG2 were cultivated in Eagles MEM
+
Earle's BSS (PAN, Cat# PO4-08510), 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA
(PAN Cat# P08-32100) and 1 mM sodium pyruvate. JHH1, JHH5, JHH6 and OZ were cultivated in Williams'E (PAN Cat# PO4-29050), 10 % FCS and 2 mM L-glutamine.
was cultivated using Williams'E, 10 % FCS and 2 mM L-glutamine. HLE was cultivated in DMEM 4,5 g/L glucose, 10 % FCS and 2 mM L-glutamine. HLF was cultivated in DMEM
4,5 g/L glucose, 5 % FCS, 0.1 mM NEAA and 2 mM L-glutamine. JHH4 was cultivated in Eagles MEM + Earle's BSS, 10 % FCS and 2 mM L-glutamine. SkHepl was cultivated in Eagles MEM + Earle's BSS, 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA and 1mM
sodium pyruvate. SNU449 was cultivated using RPMI 1640 (PAN Cat# PO4-18047) 10 %
FCS and 2 mM L-glutamine. Cells were seeded in the respective media overnight at a density of 5,000 cells per well in 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, logarithmic dilutions of 4 1 c-B starting from 27[EM down to 270pM were added and incubated for 72, 120 and 148 hours, respectively. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 dye (2[Eg/ml, Sigma Cat# B2261). Data shown are means + SD
from triplicate wells based on the analysis of 9 images per well relative to the DMSO control.
None of the cell lines tested showed a significant increase in proliferation upon treatment with 41c-B directly at the depicted time points. Results are shown in Figure 6A.
Example 58 Treatment with an active form of the compounds of the present invention (compound 41c-A) does not induce enhanced tumor cell proliferation in cell lines originating from hepatocellular carcinoma and cholangiocarcinoma Cell lines derived from hepatocellular carcinoma and cholangiocarcinoma (EGI1) were maintained and tested in the following media: Huh7 and EGI1 were cultured in DMEM 4,5 g/L glucose (Gibco, Cat# 31966-021), 10 % FCS (GIBCO, Cat# 10500-064 Lot 07G3690K), 2 mM L-glutamine (Thermo Fischer, Cat# 25030081), 1mM sodium pyruvate (GIBCO Cat# 11360-039). Hep3B and HepG2 were cultivated in Eagles MEM +
Earle's BSS (PAN, Cat# PO4-08510), 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA (PAN Cat#
P08-32100) and 1 mM sodium pyruvate. JHH2 was cultivated using Williams'E, 10 % FCS
and 2 mM L-glutamine. HLF was cultivated in DMEM 4,5 g/L glucose, 5 % FCS, 0.1 mM
NEAA and 2 mM L-glutamine. SkHep 1 was cultivated in Eagles MEM + Earle's BSS, 10 % FCS, 2 mM L-glutamine, 0.1 mM NEAA and 1mM sodium pyruvate. Cells were seeded in the respective media overnight at a density of 5,000 cells per well in 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904).
The next day, logarithmic dilutions of compound 41c-A starting from 27[EM down to 270pM were added and incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 dye (2[Eg/ml, Sigma Cat# B2261). Data shown are means + SD from triplicate wells based on the analysis of 9 images per well relative to the DMSO control.
None of the cell lines tested showed a significant increase in proliferation upon treatment with 41c-A directly after 72 hrs. Results are shown in Figure 6B.
Example 59 Treatment of tumor Cells with an active form of the compounds of the present invention (compound 41c-B) in the presence of peripheral blood results in inhibition of proliferation of tumor cells.
Heparinized whole blood of 3 different donors was diluted 1:1 in RPMI media (PAN Cat.
# PO4-18047) plus 10% FCS (GIBCO Cat# 10500-064, lot 07G3690K) and incubated at 37 C and 5% CO2 for 24hrs with 2.7[EM compound 4 1c-B. Supernatant was harvested and centrifuged at 600x g for 8 minutes to remove residual leukocytes, platelets and erythrocytes. Supernatants were stored at -80 C until further use and thawed gently at room temperature prior to addition to the cell lines. Cell lines Huh7, JHH2, HLE, HLF, JHH4, Hep3B, HepG2, JHH1, EGI1, JHH5, JHH6, OZ, SkHep 1, 5NU449 were seeded in 100[d of the respective media (as described in Example 57) overnight at a density of 5,000 cells per well 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, 100[d of the whole blood supernatants were added to the cell lines. As controls, supernatant of whole blood without addition of 4 lc-B
compound ("whole blood w/o") or plain RPMI media plus FCS ("media CTRL") was added.
Cell lines were incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 (2[Eg/ml, Sigma Cat# B2261) and viability was assessed by additional detection of Propidium Iodine (PI, 1 Kg/ml, Sigma Cat#
P4864).
Data shown are means + SD from triplicate wells based on the analysis of 9 images per well.
Results are shown in Figures 7A and 7B. For some cell lines (SNU449, JHH2 and SkHep) the addition of supernatant of non-stimulated whole blood induced proliferation above the media control level, while others responded with reduced proliferation (OZ, JHH1, HepG2, JHH4, JHH6, JHH5 and EGI1). However, treatment with supernatants derived from whole blood incubated with 4 lc-B resulted in reduced cell counts in all cases tested compared to the respective "whole blood w/o" controls. The reduced cell counts were mainly attributed to a stop in proliferation, and only the cell lines JHH2, JHH4, JHH6, Hep3B
and EGI1 did undergo cell death as determined by considerable PI positivity (data not shown).
Example 60 Factors released in peripheral blood upon treatment with an active form of the compounds of the present invention (compound 41c-A) results in inhibition of proliferation in tumor cell lines Heparinized whole blood of 2 different donors was diluted 1:1 in RPMI media (PAN Cat.
# PO4-18047) plus 10% FCS (GIBCO Cat# 10500-064, lot 07G3690K) and incubated at 37 C and 5% CO2 for 24hrs with 2.7[EM compound 4 lc-A. Supernatant was harvested and centrifuged at 600x g for 8 minutes to remove residual leukocytes, platelets and erythrocytes. Supernatants were stored at -80 C until further use and thawed gently at room temperature prior to addition to the cell lines. Cell lines Huh7, JHH2, HLF, Hep3B, HepG2, EGI1 and SkHep 1 were seeded in 100[d of the respective media (as described in Figure 6) overnight at a density of 5,000 cells per well 96 well flat clear bottom black polystyrene TC-treated microplates (Corning, Cat #3904). The next day, 100 1 of the whole blood supernatants were added to the cell lines. As controls, supernatant of whole blood without addition of 41c-A compound ("whole blood w/o") or plain RPMI
media plus FCS ("media CTRL") was added. Cell lines were incubated for 72 hours. Tumor cell counts were determined using Perkin Elmer Operetta Imaging System and Harmony Software by counting nuclei stained for 20 minutes in full media using Hoechst33342 (2[Eg/ml, Sigma Cat# B2261) and viability was assessed by additional detection of Propidium Iodine (PI, 1 lag/ml, Sigma Cat# P4864). Data shown are means + SD
from triplicate wells based on the analysis of 9 images per well.
Results are shown in Figure 7C- treatment with supernatants derived from whole blood incubated with 41c-A resulted in reduced or stable cell counts in all cases tested compared to the respective "whole blood w/o" controls. Only in Hep3B, donor #5 supernatant increased proliferation of this cell line, while supernatant from donor #4 had no impact on tumor cell proliferation.
Example 61 Single dose PK study in Male Wister-Han Rats The single dose PK in Male Wister-Han Rats was performed to assess pharmacokinetic properties of tested compounds. Two groups of animals were dosed via Gavage (POE) of the respective compound. Blood samples (approximately 20 [iL) were collected via Jugular vein or an alternate site at 15 mm, 30 min, 1H, 2 h, 4 h, 7 h and 24 h post-dose groups. Blood samples were placed into tubes containing EDTA-K2 anticoagulant and centrifuged at 5000 rpm for 6 min at 4 C to separate plasma from the samples. After centrifugation, the resulting plasma was transferred to clean tubes for bioanalysis of both prodrug and active form on LC/MS/MS. In the groups that prodrug were dosed, the concentration of prodrugs in the plasma samples was under the detection limit. The "tested compound" in Table 8 was used as the internal standard for testing the metabolite (active form) of "dose compound" in vivo. The pharmacokinetic parameters were calculated using non-compartmental module of WinNonlin0 Professional 6.2.
The peak concentration (C.) was recorded directly from experimental observations.
The area under the plasma concentration-time curve (AUCort) was calculated using the linear trapezoidal rule up to the last detectable concentration.
Cmax and AUCo-last are two critical PK parameters related to the in vivo efficacy of the tested compound. Compounds with higher Cmax and AUCo-last will lead to the better in vivo efficacy. Results of PK parameters following oral administration of active forms and competitor compounds are given in Table 7. The PK parameters of prodrugs are tabulated in Table 8.
Following oral administration of prodrugs, the active forms were observed in plasma and therefore tested. The exemplified prodrugs of present invention (Example 41-B, 42-A, 42-B, 43-A, 45-A and 45-B) surprisingly showed much improved Cmax (5-175 folds increase) and AUCo-iast (2.5-56 folds increase) comparing with reference compounds (G59620, S-2 and S-3) and compounds mentioned in present invention (Compound 41c-A, 41c-B and 43e-A) which are all active forms. The results clearly demonstrated the unexpected superiority of prodrugs over active forms on PK parameters which led to better in vivo efficacy.
Table 7. The mean plasma concentration and PK parameters of active forms after 5 mg/kg oral dosing Dose Compound compound 41c-A
Time (h) Mean plasma concentration (nM) 0.25 56.3 9.49 8.89 16.75 0.5 33.2 16.74 9.99 27.48 1 83.4 19.33 10.16 32.33 2 136 24.89 8.40 27.34 4 16.7 47.55 11.54 27.38 8* 9.49 52.72 8.17 18.02 24 ND 4.90 ND 5.60 Cmax (nM) 164 52.72 11.54 32.33 AUCo_iast 316 748 95 242.5 (nM=h) Dose Compound Compound Compound Compound compound 41c-B 43e-A 45e-A 45e-B
Time (h) Mean plasma concentration (nM) 0.25 3.41 12.60 64.6 42.8 0.5 0.75 15.22 80.0 52.2 1 2.04 13.01 58.1 37.6 2 5.46 11.98 42.5 24.2 4 2.52 8.20 77.8 53.9 8* 1.21 6.31 34.6 29 24 ND ND 8.6 5.7 Cmax (nM) 5.46 15.22 80.0 53.9 AUCo_iast 767 568 55.8 77 (nM=h) * 7 hrs for Compound 41c-A, Compound 41c-B and Compound 43e-A
Table 8. PK parameters of prodrugs after 5 mg/kg oral dosing Dose compound Tested compound C. (nM) AUCo-iast (nM=h) Example 41-B Compound 41c-A 1315 3658 Example 42-A Compound 41c-A 1742 4867 Example 42-B Compound 41c-B 956 3148 Example 43-A Compound 43e-A 77 229 Example 45-A Compound 45e-B 922 1914 Example 45-B Compound 45e-A 1436 2619 Example 62 LYSA solubility study LYSA study is used to determine the aqueous solubility of tested compounds.
Samples were prepared in duplicate from 10 mM DMSO stock solution. After evaporation of DMSO with a centrifugal vacuum evaporator, the compounds were dissolved in 0.05 M
phosphate buffer (pH 6.5), stirred for one hour and shaken for two hours.
After one night, the solutions were filtered using a microtiter filter plate. Then the filtrate and its 1/10 dilution were analyzed by HPLC-UV. In addition, a four-point calibration curve was prepared from the 10 mM stock solutions and used for the solubility determination of the compounds. The results were in [tg/mL. In case the percentage of sample measured in solution after evaporation divided by the calculated maximum of sample amount was bigger than 80%, the solubility was reported as bigger than this value.
Results of LYSA were shown in Table 9. It was clear that the solubility of active forms were surprisingly improved by 10 to over 200 folds when converted to various prodrugs.
Table 9. Solubility data of particular compounds LYSA of LYSA of Active Corresponding Active Prodrugs Prodrugs Forms Forms ( g/mL) ( g/mL) Example 1 290 Compound le 21 Example 1-A 315 Compound le-A 56 Example 1-B 200 Compound le-B 50 Example 2 615 Compound le 21 Example 2-A >600 Compound le-B 50 Example 2-B >590 Compound le-A 56 Example 3 240 Compound le 21 Example 4 695 Compound le 21 Example 5 >595 Compound le 21 Example 6 140 Compound le 21 Example 7 615 Compound le 21 Example 8 620 Compound le 21 Example 9 >520 Compound le 21 Example 10 120 Compound le 21 Example 11 >618 Compound le 21 Example 12 120 Compound le 21 Example 13 155 Compound le 21 Example 14 225 Compound le 21 Example 15 405 Compound le 21 Example 16 205 Compound le 21 Example 17 190 Compound le 21 Example 25 >670 Compound le 21 Example 26 >690 Compound le 21 Example 27 >380 Compound le 21 Example 28 695 Compound le 21 Example 29 395 Compound le 21 Example 32 125 Compound le 21 Example 36-A 168 Compound 36g-A 6 Example 36-B 209 Compound 36g-B 11 Example 41-A 260 Compound 41c-B 5 Example 41-B 250 Compound 41c-A 1 Example 42-A 225 Compound 41c-A 1 Example 42-B 335 Compound 41c-B 5 Example 43-A 203 Compound 43e-A 13 Example 43-B 170 Compound 43e-B 13 Example 45 172 Compound 45e 152 Example 45-A >560 Compound 45e-A or 90 or 115 Compound 45e-B
Example 45-B 420 Compound 45e-B 115 or 90 Or Compound 45e-A
Example 46-A 205 Compound 45e-A 90 or 115 Or Compound 45e-B
Example 46-B >580 Compound 45e-B 115 or 90 Or Compound 45e-A
Example 47-A 154 Compound 47e-A or <1.0 or <1.0 Compound 47e-B
Example 47-B 128 Compound 47e-B or <1.0 or <1.0 Compound 47e-A
Example 48-A 305 Compound 47e-A or <1.0 or <1.0 Compound 47e-B
Example 48-B 275 Compound 47e-B or <1.0 or <1.0 Compound 47e-A
Example 63 Portal Vein Study The objective of this study was to understand whether pro drug remains unchanged as it was absorbed through the intestine into the portal circulation and demonstrate the primary site of conversion.
Surgical procedure for Portal vein cannulation (PVC) and Carotid artery cannulation (CAC) Surgery was performed under pentobarbital /isoflurane anesthesia. Briefly, after disinfecting the abdominal area with betadine and 70% isopropyl alcohol, a small abdominal mid-line incision was made. The cecum was pulled out and mesenteric vein was identified and isolated for about 5 mm vessel. A loose ligature was placed proximally and distal end of the vein was ligated. Make a small incision (just enough to allow the insertion of the catheter) on isolated vein and insert the PU catheter towards liver for appropriate length. The catheter was secured in place by tying the loose ligature around the cannulated vessel. The cecum was replaced into abdominal cavity. A hole was made in the right abdominal wall to make the end of catheter pass freely. The catheter was secured by suture on the abdominal wall. The abdominal muscle incision was closed with suture. A
small incision was made in the scapular area to serve as the exit site of the catheter. The catheter was subcutaneously tunneled and exteriorized through the scapular incision. A
fixed suture was placed in the scapular region. The patency of the catheter was checked and then exteriorized from the subcutaneous space to the dorsal neck region. After gently wiping the area, the abdominal cavity was sutured. The left carotid artery was then cannulated by inserting a PESO catheter. Both the exteriorized catheters were tied firmly on the dorsal neck region and fixed. The animals was then allowed to recover in its cage and used for study at least 3 days after surgery. All catheters were flushed once daily with heparinized saline to maintain patency.
Oral PK study in PVC/CAC dual cannulated rat Animals were fasted overnight (n=3) and administered vial oral gavage (10mg/kg, 10mL/kg). Blood samples (60 [LL) were collected simultaneously from the portal and carotid artery catheters at 0.083, 0.25, 0.5, 1, 2, 4, 7, 24h. All blood samples will be transferred into microcentrifuge tubes containing 2 [LI., of K2EDTA (0.5M) as anti-coagulant and placed on wet ice. Then blood samples will be processed for plasma by centrifugation at approximately 4 C, 3000g within half an hour of collection.
Plasma samples will be stored in polypropylene tubes, quick frozen over dry ice and kept at -70 10 C until LC/MS/MS analysis.
Pharmacokinetic parameters (mean SD, n= 3) of prodrugs and active forms in portal and carotid samples following oral administration of prodrugs (10 mg/kg) in portal vein cannulated rat were detected and analyzed. The test results of Example 1-B, 41-A, 41-B, 42-A and 43-A were summarized below.
Table 10. Pharmacokinetic parameters of Example 41-A and its corresponding active form Compound 41c-B in portal and carotid samples following oral administration of Example 41-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 41-A
Corresponding Active Form Compound 41c-B
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.14 0.4 0.19 0.42 C. (nM) 9703 2223 210 2185 AUC0_2 (nM=h) 2188 2246 114 2108 AUCactive/AUC 1 tota, 51% 95%
Table 11. Pharmacokinetic parameters of Example 43-A and its corresponding active form Compound 43e-A in portal and carotid samples following oral administration of Example 43-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 43-A
Corresponding Active Form Compound 43e-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form Tmax (h) 0.28 0.33 0.22 0.28 C. (nM) 4110 818 191 691 AUC0_2 (nM=h) 2067 679 124 564 AUCactive/AUC 1 tota, 25% 82%
Table 12. Pharmacokinetic parameters of Example 1-B and its corresponding active form Compound le-A in portal and systemic samples following oral administration of Example 1-B (10 mg/kg) in portal vein cannulated rat Prodrug Example 1-B
Corresponding Active Form Compound le-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.083 0.25 0.083 0.5 C. (nM) 670 192 70 174 AUC0_2 (nM=h) 266 164 40 184 AUCactive/AUC 1 tota, 38% 82%
Table 13. Pharmacokinetic parameters of Example 42-A and its corresponding active form Compound 41c-A in portal and carotid samples following oral administration of Example 42-A (10 mg/kg) in portal vein cannulated rat Prodrug Example 42-A
Corresponding Active Form Compound 41c-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form T. (h) 0.19 0.42 0.22 0.36 C. (nM) 8917 3162 286 3326 AUC0_2 (nM=h) 3461 3199 286 3326 AUCactive/AUC 1 tota, 48% 96%
Table 14. Pharmacokinetic parameters of Example 41-B and its corresponding active form Compound 41c-A in portal and carotid samples following oral administration of Example 41-B (10 mg/kg) in portal vein cannulated rat Prodrug Example 41-B
Corresponding Active Form Compound 41c-A
Portal sampling Carotid sampling PK parameter prodrug active form prodrug active form Triax (h) 0.19 0.5 0.25 0.5 C. (nM) 7068 3315 29.6 3432 AUC0_2 (nM=h) 1444 3211 22.5 3301 AUCactive/AUC
tota, 69% 99%
Based on the above results, it was concluded that the primary site of conversion of prodrug was liver rather than intestine, because AUCactive/AUC I was higher in sampling tota, from carotid artery compared to AUCactive/AUCtota, in sampling from portal vein.
Claims (38)
1. A compound of formula (I), N H 2 ....¨R3 N------ N
H N 1 > __ O
" 'N
0=S N
1 1 \ 2 R
R (I), wherein R1 is Ci_olkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_olkyl;
R3 is -NR4R5, wherein R4 is Ci_olkyl or Ci_olkoxyCi_olkyl;
R5 is (Ci_6a1ky1)2NCOOCi_6a1ky1, Ci_6a1koxyCi_6a1ky1, Ci-6alkoxycarbonyl(Ci_olkyl)aminoCi_olkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6a1ky1, Ci_olkoxycarbonylCi_olkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_olkylcarbonyl(Ci_ 6alkyl)aminoCi_6a1ky1 or pyrrolidinylcarbamoyloxyCi_olkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3,3-dimethylpyrro lidine-l-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
- 196 -6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3- fluoro-3-methyl-pyrro lidine-l-carb ony1)-2-(propylsulfonimidoyl)purin-8- one;
and their enantiomers or diastereomers are excluded.
H N 1 > __ O
" 'N
0=S N
1 1 \ 2 R
R (I), wherein R1 is Ci_olkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by one, two or three substituents independently selected from halogen and Ci_olkyl;
R3 is -NR4R5, wherein R4 is Ci_olkyl or Ci_olkoxyCi_olkyl;
R5 is (Ci_6a1ky1)2NCOOCi_6a1ky1, Ci_6a1koxyCi_6a1ky1, Ci-6alkoxycarbonyl(Ci_olkyl)aminoCi_olkyl, Ci_ 6alkoxycarbonyl(phenyl)Ci_6a1ky1, Ci_olkoxycarbonylCi_olkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_olkylcarbonyl(Ci_ 6alkyl)aminoCi_6a1ky1 or pyrrolidinylcarbamoyloxyCi_olkyl; or R4 and R5 together with the nitrogen they are attached to form a heterocyclyl;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof;
for use in the treatment or prophylaxis of liver cancer;
with the proviso that 6-amino-9-benzy1-2-(propylsulfonimidoy1)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-amino-9-benzy1-7-(piperidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(morpholine-4-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3,3-dimethylpyrro lidine-l-carb ony1)-2-(propylsulfonimidoyl)purin-8-one;
ethyl 1-[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]pyrrolidine-2-carboxylate;
6-amino-7-(2-azaspiro[3.3]heptane-2-carbony1)-9-benzy1-2-(propylsulfonimidoyl)purin-8-one;
- 196 -6-amino-9-benzy1-7-(2-oxa-6-azaspiro[3.3]heptane-6-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-benzy1-7-(3,3-difluoropyrrolidine-1-carbony1)-2-(propylsulfonimidoyl)purin-8-one;
6-amino-9-b enzy1-7-(3- fluoro-3-methyl-pyrro lidine-l-carb ony1)-2-(propylsulfonimidoyl)purin-8- one;
and their enantiomers or diastereomers are excluded.
2. The compound for use according to claim 1, wherein Ri is Ci_olkyl;
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
piperazinyl substituted by Ci_olkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is Ci_olkyl or Ci_olkoxyCi_olkyl;
R5 is (Ci_6a1ky1)2NCOOCi_6a1ky1, Ci_olkoxyCi_olkyl, Ci-6alkoxycarbonyl(Ci_olkyl)aminoCi_olkyl, Ci_ 6alkoxycarbonyl(phenyl)C1-6alkyl, Ci_olkoxycarbonylCi_olkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_olkylcarbonyl(Ci_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyCi_olkyl.
R2 is benzyl, said benzyl being unsubstituted or substituted by halogen or Ci_6alkyl;
R3 is azetidinyl;
piperazinyl substituted by Ci_olkyl;
piperidinyl substituted by piperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is Ci_olkyl or Ci_olkoxyCi_olkyl;
R5 is (Ci_6a1ky1)2NCOOCi_6a1ky1, Ci_olkoxyCi_olkyl, Ci-6alkoxycarbonyl(Ci_olkyl)aminoCi_olkyl, Ci_ 6alkoxycarbonyl(phenyl)C1-6alkyl, Ci_olkoxycarbonylCi_olkyl, Ci_ 6alkoxycarbonyloxyCi_6alkyl, Ci_6alkyl, Ci_olkylcarbonyl(Ci_ 6alkyl)aminoCi_6alkyl or pyrrolidinylcarbamoyloxyCi_olkyl.
3. The compound for use according to claim 1 or 2, wherein Ri is ethyl or propyl;
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl.
R2 is benzyl, bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl;
R3 is azetidinyl;
4-methylpiperazinyl;
piperidinylpiperidinyl;
pyrrolidinyl; or -NR4R5, wherein R4 is methyl, ethyl, propyl or methoxyethyl;
R5 is acetyl(methyl)aminoethyl, butyl, butyl(methyl)carbamoyloxyethyl, diethylcarbamoyloxyethyl, ethoxycarbonyl(methyl)aminoethyl, ethoxycarbonylethyl, ethoxycarbonylisobutyl, ethoxycarbonylisopentyl, ethoxycarbonylmethyl, ethoxycarbonyloxyethyl, ethoxycarbonyl(phenyl)ethyl, ethyl, isobutyl, isopropoxycarbonylisopentyl, isopropoxycarbonyl(phenyl)ethyl, isopropyl, methoxycarbonyl(methyl)aminoethyl, methoxyethyl, methoxypropyl, propyl, propyl(methyl)carbamoyloxyethyl, pyrrolidinylcarbamoyloxyethyl, tert-butoxycarbonyl(methyl)aminoethyl, tert-butoxycarbonylethyl, tert-butoxycarbonylisopentyl or tert-butoxycarbonyl(phenyl)ethyl.
4. The compound for use according to claim 3, wherein R3 is azetidinyl, 4-methylpiperazinyl, piperidinylpiperidinyl, pyrrolidinyl, acetyl(methyl)aminoethyl(methyl)amino, bis(methoxyethyl)amino, butyl(ethyl)amino, butyl(methyl)amino, butyl(methyl)carbamoyloxyethyl(methyl)amino, diethylcarbamoyloxyethyl(methyl)amino, ethoxycarbonyl(methyl)aminoethyl(methyl)amino, ethoxycarbonylethyl(methyl)amino, ethoxycarbonylisobutyl(methyl)amino, ethoxycarbonylisopentyl(methyl)amino, ethoxycarbonylmethyl(methyl)amino, ethoxycarbonyloxyethyl(methyl)amino, ethoxycarbonyl(phenyl)ethyl(methyl)amino, ethyl(methyl)amino, isobutyl(methyl)amino, isopropoxycarbonylisopentyl(methyl)amino, isopropoxycarbonyl(phenyl)ethyl(methyl)amino, isopropyl(methyl)amino, methoxycarbonyl(methyl)aminoethyl(methyl)amino, methoxyethyl(ethyl)amino, methoxyethyl(methyl)amino, methoxyethyl(propyl)amino, methoxypropyl(methyl)amino, propyl(ethyl)amino, propyl(methyl)amino, propyl(methyl)carbamoyloxyethyl(methyl)amino, pyrrolidinylcarbamoyloxyethyl(methyl)amino, tert-butoxycarbonyl(methyl)aminoethyl(methyl)amino, tert-butoxycarbonylethyl(methyl)amino, tert-butoxycarbonylisopentyl(methyl)amino or tert-butoxycarbonyl(phenyl)ethyl(methyl)amino.
5. The compound for use according to any one of claims 1 to 4, wherein R1 is ethyl.
6. The compound for use according to claim 1 or 2, wherein R2 is benzyl substituted by halogen or Ci_olkyl.
7. The compound for use according to any one of claims 2 to 6, wherein R2 is bromobenzyl, chlorobenzyl, fluorobenzyl or methylbenzyl.
8. The compound for use according to claim 7, wherein R2 is bromobenzyl, chlorobenzyl or fluorobenzyl.
9. The compound for use according to claim 1 or 2, wherein R3 is -NR4R5, wherein R4 is Ci_olkyl, R5 is Cl_olkyl.
10. The compound for use according to claim 9, wherein R3 is propyl(methyl)amino or ethyl(methyl)amino.
11. The compound for use according to any one of claims 1, 2, 6 and 9, wherein R1 is Ci_olkyl;
R2 is benzyl, said benzyl being substituted by halogen or Ci_olkyl;
R3 is -NR4R5, wherein R4 is Ci_olkyl, R5 is Ci_olkyl.
R2 is benzyl, said benzyl being substituted by halogen or Ci_olkyl;
R3 is -NR4R5, wherein R4 is Ci_olkyl, R5 is Ci_olkyl.
12. The compound for use according to claim 11, wherein R1 is ethyl;
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino.
R2 is methylbenzyl, bromobenzyl, chlorobenzyl or fluorobenzyl;
R3 is propyl(methyl)amino or ethyl(methyl)amino.
13. A compound for use in the treatment or prophylaxis of liver cancer selected from:
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino- 9-b enzy1-7- (4-methylpip erazine- 1 -curb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (25)-2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (25)-2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3 -phenyl-propanoate;
Isopropyl (25)-2- [[6-amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3 -phenyl-propanoate;
tert-Butyl (25)-2- [[6-amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2- [Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
2- [[6-Amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-butyl-N-methyl-carbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl pyrrolidine-l-carboxylate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-methyl-N-propyl-carbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N,N-diethylcarbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl ethyl carbonate;
- 201 -6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2-[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
- 202 -6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)nethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
- 203 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
6-Amino-9-benzyl-N-methy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-ethy1-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzy1-7-[4-(1-piperidyl)piperidine-1-carbony1]-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-ethyl-N-(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-butyl-N-ethy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-(2-methoxyethyl)-8-oxo-N-propy1-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N,N-bis(2-methoxyethyl)-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-7-(azetidine-1-carbony1)-9-benzyl-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-isopropyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino- 9-b enzy1-7- (4-methylpip erazine- 1 -curb ony1)-2-(propylsulfonimidoyl)purin-8-one;
6-Amino-9-benzyl-N-(3-methoxypropy1)-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-9-benzyl-N-isobutyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Ethyl 2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]acetate;
Ethyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]propanoate;
tert-Butyl 3-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]propanoate;
tert-Butyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Isopropyl (25)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (2S)-2-[[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-methyl-butanoate;
Ethyl (25)-2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-4-methyl-pentanoate;
Ethyl (25)-2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3 -phenyl-propanoate;
Isopropyl (25)-2- [[6-amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3 -phenyl-propanoate;
tert-Butyl (25)-2- [[6-amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]-3-phenyl-propanoate;
N- [2- [Acetyl(methyl)amino]ethy1]-6-amino-9-benzyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
Methyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
tert-Butyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
Ethyl N- [2- [[6-amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbony1]-methyl-amino]ethy1]-N-methyl-carbamate;
2- [[6-Amino-9-benzyl- 8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-butyl-N-methyl-carbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl pyrrolidine-l-carboxylate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N-methyl-N-propyl-carbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl N,N-diethylcarbamate;
2- [[6-Amino-9-benzy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carbonyl]-methyl-amino]ethyl ethyl carbonate;
- 201 -6-Amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-amino-N-buty1-9-[(4-chlorophenyl)methy1]-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-methy1-8-oxo-N-propy1-2[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-2-[S(R)-propylsulfonimidoy1]-9-(p-tolylmethyl)-7-(pyrrolidine-1-carbonyl)purin-8-one;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(S)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-(2-methoxyethyl)-N-methy1-8-oxo-2-[S(R)-propylsulfonimidoyl]-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-butyl-N-methy1-8-oxo-2-(propylsulfonimidoy1)-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-2-[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
- 202 -6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-N-methy1-8-oxo-N-propy1-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-ethylsulfonimidoy1]-N-methyl-8-oxo-9-(p-tolylmethyl)purine-7-carboxamide;
6-Amino-2-[S(S)ethylsulfonimidoy1]-9-[(4-fluorophenyOmethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)ethylsulfonimidoy1]-9-[(4-fluorophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-N-ethy1-2-(ethylsulfonimidoy1)-9-[(4-fluorophenyl)methyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(S)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-N-ethy1-2-[S(R)-(ethylsulfonimidoy1)]-9-[(4-fluorophenyl)methyl]-N-methy1-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(R)-ethylsulfonimidoy1]-9-[(4-bromophenyl)methy1]-N-methy1-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-2-[S(S)-ethylsulfonimidoy1]-9-[(4-bromophenyl)nethyl]-N-methyl-8-oxo-N-propyl-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
- 203 -6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
14. The compound for use according to claim 13, selected from:
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoyl)]-N-methy1-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2-[S(R)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-(ethylsulfonimidoy1)-N-methyl-8-oxo-purine-7-carboxamide;
6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(S)-(ethylsulfonimidoyl)]-N-methy1-8-oxo-purine-7-carboxamide; and 6-Amino-9-[(4-bromophenyl)methy1]-N-ethy1-2-[S(R)-(ethylsulfonimidoyl)]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
15. The compound for use according to claim 13, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methy1]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide;
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
or pharmaceutically acceptable salt, enantiomer or diastereomer thereof.
16. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of claims 1 to 15, wherein the liver cancer is hepatocellular carcinoma, hepatoma, cholangiocarcinoma, hepatoblastoma, hepatic carcinoma, hepatic angiosarcoma, or metastatic liver cancer.
17. The compound or pharmaceutically acceptable salt, enantiomer or diastereomer for use according to any one of claims 1 to 15, wherein the liver cancer is hepatocellular carcinoma.
18. A pharmaceutical composition or medicament comprising a compound according to any one of claims 1 to 15 and a therapeutically inert carrier, for use in the treatment or prophylaxis of liver cancer.
19. The use of a compound according to any one of claims 1 to 15 for the preparation of a medicament for the treatment or prophylaxis of liver cancer.
20. A method for the treatment or prophylaxis of liver cancer., which method comprises administering a therapeutically effective amount of a compound as defined in any one of claims 1 to 15.
21. A compound as defined in any one of claims 1 to 15, or a pharmaceutical composition or a medicament comprising such compound, for use in a) the treatment or prophylaxis of liver cancer in combination with an antagonistic PD1 antibody or antagonistic PD-L1 antibody, or b) the treatment of a patient suffering from liver cancer in combination with an antagonistic PD1 antibody or antagonistic PD-L1 antibody.
22. A compound as defined in any one of claims 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-L1 antibody.
23. Use of a compound as defined in any one of claims 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-L1 antibody.
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an antagonistic PD1 antibody or antagonistic PD-L1 antibody.
24. The compound, composition, medicament or use, according to any one of claims 21 to 23, wherein the treatment is in combination with an antagonistic PD1 antibody.
25. The compound, composition, medicament or use, according to claim 24, wherein the antagonistic PD1 antibody is nivolumab or pemprolizumab.
26. The compound, composition, medicament or use, according to claim 25, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methyl]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide
27. The compound, composition, medicament or use, according to claim 24, wherein the antagonistic PD1 antibody comprises a heavy chain variable domain VH with an amino acid sequence of SEQ ID NO: 5 and a light chain variable domain VL with an amino acid sequence of SEQ ID NO:6.
28. The compound, composition, medicament or use, according to claim 26, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methyl]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide.
29. The compound, composition, medicament or use, according to any one of claims 21 to 23, wherein the treatment is in combination with an antagonistic PD-L1 antibody.
30. The compound, composition, medicament or use, according to claim 29,wherein the antagonistic PD-L1 antibody used in the combination therapy is atezolizumab or durvalumab or avelumab (in one preferred embodiment atezolizumab)
31. The compound, composition, medicament or use, according to claim 30, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methyl]-N-ethy1-2[S(S)-ethylsulfonimidoyl]-N-methyl-8-oxo-purine-7-carboxamide.
32. The compound, composition, medicament or use according to any one of claims 21 to 31 wherein additionally an anti-angiogenic agent is used in the combination therapy
33. The compound, composition, medicament or use according to any one of claims 21 to 31 wherein additionally an anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab) is used in the combination therapy.
34. A compound as defined in any one of claims 1 to 15, or a pharmaceutical composition or a medicament comprising such compound, for use in a) the treatment or prophylaxis of liver cancer in combination with an anti-angiogenic agent, or b) the treatment of a patient suffering from liver cancer in combination with an anti-angiogenic agent.
35. A compound as defined in any one of claims 1 to 15, or a pharmaceutical composition or a medicament comprising such compound for use in the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
36. Use of a compound as defined in any one of claims 1 to 15;
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
for the preparation of a medicament for the treatment or prophylaxis of liver cancer wherein the treatment is in combination with an anti-angiogenic agent.
37. The compound, composition, medicament or use according to any one of claims 34 to 36 wherein the anti-angiogenic agent selected from is sorafenib, regorafenib, sunitinib or bevacizumab (in one preferred embodiment the anti-angiogenic agent is sorafenib; in one preferred embodiment the anti-angiogenic agent is bevacizumab).
38. The compound, composition, medicament or use, according to claim 37, wherein the compound is 6-Amino-9-[(4-chlorophenyl)methyl]-N-ethy1-2[S(S)-ethylsulfonimidoy1]-N-methy1-8-oxo-purine-7-carboxamide.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CNPCT/CN2018/077501 | 2018-02-28 | ||
CN2018077501 | 2018-02-28 | ||
PCT/EP2019/054729 WO2019166432A1 (en) | 2018-02-28 | 2019-02-26 | 7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer |
Publications (1)
Publication Number | Publication Date |
---|---|
CA3091950A1 true CA3091950A1 (en) | 2019-09-06 |
Family
ID=65685310
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CA3091950A Pending CA3091950A1 (en) | 2018-02-28 | 2019-02-26 | 7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer |
Country Status (19)
Country | Link |
---|---|
EP (1) | EP3758707A1 (en) |
JP (1) | JP7089596B2 (en) |
KR (1) | KR20200128414A (en) |
CN (1) | CN111801100B (en) |
AR (1) | AR114419A1 (en) |
AU (1) | AU2019228654A1 (en) |
BR (1) | BR112020016509A2 (en) |
CA (1) | CA3091950A1 (en) |
CL (1) | CL2020002139A1 (en) |
CO (1) | CO2020010306A2 (en) |
IL (1) | IL276817A (en) |
MA (1) | MA52412A (en) |
MX (1) | MX2020008746A (en) |
PE (1) | PE20211456A1 (en) |
PH (1) | PH12020551343A1 (en) |
RU (1) | RU2020131012A (en) |
SG (1) | SG11202008291XA (en) |
TW (1) | TW202003518A (en) |
WO (1) | WO2019166432A1 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112420196A (en) * | 2020-11-20 | 2021-02-26 | 长沙市弘源心血管健康研究院 | Prediction method and system for survival rate of acute myocardial infarction patient within 5 years |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JOP20170169A1 (en) | 2016-08-29 | 2019-01-30 | Novartis Ag | Fused tricyclic pyridazinone compounds useful to treat orthomyxovirus infections |
KR102409595B1 (en) | 2020-06-29 | 2022-06-17 | 한국과학기술연구원 | Novel purinone derivatives as protein kinase CSF-1R inhibitor |
WO2024013205A1 (en) * | 2022-07-14 | 2024-01-18 | F. Hoffmann-La Roche Ag | Phosphorylpurinone compounds for the treatment of cancer |
Family Cites Families (11)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP4189048B2 (en) * | 1997-12-26 | 2008-12-03 | 大日本住友製薬株式会社 | Heterocyclic compounds |
ES2623794T3 (en) | 2008-12-09 | 2017-07-12 | Gilead Sciences, Inc. | Intermediates for the preparation of toll receptor modulators |
CA2760766A1 (en) * | 2009-05-21 | 2010-11-25 | Nicholas James Bennett | Novel pyrimidine derivatives and their use in the treatment of cancer and further diseases |
IN2012DN02984A (en) * | 2009-10-22 | 2015-07-31 | Gilead Sciences Inc | |
WO2011079016A1 (en) * | 2009-12-22 | 2011-06-30 | Gilead Sciences, Inc. | Methods of treating hbv and hcv infection |
WO2011163594A2 (en) | 2010-06-24 | 2011-12-29 | Alkermes, Inc. | Prodrugs of nh-acidic compounds: ester, carbonate, carbamate and phosphonate derivatives |
KR20130108295A (en) * | 2010-08-13 | 2013-10-02 | 베일러 리서치 인스티튜트 | Novel vaccine adjuvants based on targeting adjuvants to antibodies directly to antigen-presenting cells |
MY182353A (en) | 2015-05-08 | 2021-01-20 | Hoffmann La Roche | Novel sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of virus infection |
MA44334A (en) | 2015-10-29 | 2018-09-05 | Novartis Ag | ANTIBODY CONJUGATES INCLUDING A TOLL-TYPE RECEPTOR AGONIST |
CN109641904B (en) | 2016-08-29 | 2022-01-28 | 豪夫迈·罗氏有限公司 | 7-substituted sulfoxy-purinone compounds for the treatment and prevention of viral infections |
EP3970750A1 (en) * | 2016-09-13 | 2022-03-23 | F. Hoffmann-La Roche AG | Combined treatment with a tlr7 agonist and an hbv capsid assembly inhibitor |
-
2019
- 2019-02-26 WO PCT/EP2019/054729 patent/WO2019166432A1/en unknown
- 2019-02-26 PE PE2020001228A patent/PE20211456A1/en unknown
- 2019-02-26 BR BR112020016509-3A patent/BR112020016509A2/en not_active Application Discontinuation
- 2019-02-26 CA CA3091950A patent/CA3091950A1/en active Pending
- 2019-02-26 MX MX2020008746A patent/MX2020008746A/en unknown
- 2019-02-26 AU AU2019228654A patent/AU2019228654A1/en not_active Abandoned
- 2019-02-26 JP JP2020544794A patent/JP7089596B2/en active Active
- 2019-02-26 EP EP19708981.6A patent/EP3758707A1/en active Pending
- 2019-02-26 MA MA052412A patent/MA52412A/en unknown
- 2019-02-26 SG SG11202008291XA patent/SG11202008291XA/en unknown
- 2019-02-26 KR KR1020207027935A patent/KR20200128414A/en unknown
- 2019-02-26 RU RU2020131012A patent/RU2020131012A/en unknown
- 2019-02-26 CN CN201980015912.7A patent/CN111801100B/en active Active
- 2019-02-27 TW TW108106752A patent/TW202003518A/en unknown
- 2019-02-28 AR ARP190100505A patent/AR114419A1/en unknown
-
2020
- 2020-08-19 CL CL2020002139A patent/CL2020002139A1/en unknown
- 2020-08-19 IL IL276817A patent/IL276817A/en unknown
- 2020-08-21 CO CONC2020/0010306A patent/CO2020010306A2/en unknown
- 2020-08-27 PH PH12020551343A patent/PH12020551343A1/en unknown
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112420196A (en) * | 2020-11-20 | 2021-02-26 | 长沙市弘源心血管健康研究院 | Prediction method and system for survival rate of acute myocardial infarction patient within 5 years |
Also Published As
Publication number | Publication date |
---|---|
IL276817A (en) | 2020-10-29 |
EP3758707A1 (en) | 2021-01-06 |
JP2021514972A (en) | 2021-06-17 |
JP7089596B2 (en) | 2022-06-22 |
CO2020010306A2 (en) | 2020-08-31 |
PE20211456A1 (en) | 2021-08-05 |
BR112020016509A2 (en) | 2020-12-15 |
WO2019166432A1 (en) | 2019-09-06 |
CN111801100A (en) | 2020-10-20 |
CN111801100B (en) | 2023-10-24 |
KR20200128414A (en) | 2020-11-12 |
PH12020551343A1 (en) | 2021-06-21 |
AR114419A1 (en) | 2020-09-02 |
SG11202008291XA (en) | 2020-09-29 |
CL2020002139A1 (en) | 2021-01-04 |
TW202003518A (en) | 2020-01-16 |
MA52412A (en) | 2021-06-02 |
AU2019228654A1 (en) | 2020-09-03 |
RU2020131012A (en) | 2022-03-28 |
MX2020008746A (en) | 2020-09-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2021204303B2 (en) | 7-substituted sulfonimidoylpurinone compounds for the treatment and prophylaxis of virus infection | |
CA3091950A1 (en) | 7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer | |
CA2774145C (en) | Hcv protease inhibitors | |
CA3095494A1 (en) | Modulators of proteolysis and associated methods of use | |
BR112021002479A2 (en) | compound, pharmaceutical composition, method of inhibiting the activity of itk or jak3 in a population of cells, method of treating an itk or jak3 mediated disorder in an individual in need thereof, and use of a compound | |
CA3134173A1 (en) | Phosphatidylinositol 3-kinase inhibitors | |
CA3049175A1 (en) | Jak1 selective inhibitors | |
CA3133300A1 (en) | Benzodiazepine derivatives as rsv inhibitors | |
JP2012521426A (en) | Novel aminopyridine derivatives having selective inhibition of Aurora A | |
US20200268762A1 (en) | 7-substituted sulfonimidoylpurinone compounds and derivatives for the treatment and prophylaxis of liver cancer | |
TW201736391A (en) | Alkynyl nucleoside analogs as inhibitors of human rhinovirus | |
ES2962774T3 (en) | Inhibitors of human immunodeficiency virus replication | |
WO2015193229A1 (en) | Bet-protein inhibiting 1,4-dihydropyrido[3,4-b]pyrazinones with meta-substituted aromatic amino- or ether groups | |
IL296182A (en) | Inhibitors of human immunodeficiency virus replication | |
JP2023155223A (en) | Method of treatment including kras g12c inhibitors and aurora a inhibitors |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
EEER | Examination request |
Effective date: 20220816 |
|
EEER | Examination request |
Effective date: 20220816 |
|
EEER | Examination request |
Effective date: 20220816 |
|
EEER | Examination request |
Effective date: 20220816 |
|
EEER | Examination request |
Effective date: 20220816 |