CN1705675A - 亚磷酰化方法 - Google Patents
亚磷酰化方法 Download PDFInfo
- Publication number
- CN1705675A CN1705675A CNA2003801014035A CN200380101403A CN1705675A CN 1705675 A CN1705675 A CN 1705675A CN A2003801014035 A CNA2003801014035 A CN A2003801014035A CN 200380101403 A CN200380101403 A CN 200380101403A CN 1705675 A CN1705675 A CN 1705675A
- Authority
- CN
- China
- Prior art keywords
- group
- replacement
- unsubstituted
- activator
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 28
- 238000005731 phosphitylation reaction Methods 0.000 title abstract description 4
- 239000012190 activator Substances 0.000 claims abstract description 25
- 150000007530 organic bases Chemical class 0.000 claims abstract description 21
- 108091034117 Oligonucleotide Proteins 0.000 claims abstract description 19
- 239000002777 nucleoside Substances 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 12
- 125000003835 nucleoside group Chemical group 0.000 claims abstract description 12
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 11
- 150000003839 salts Chemical class 0.000 claims abstract description 5
- -1 phosphorus acylated reagent alcohol Chemical class 0.000 claims description 36
- 229910052757 nitrogen Inorganic materials 0.000 claims description 33
- 230000000903 blocking effect Effects 0.000 claims description 31
- 125000003118 aryl group Chemical group 0.000 claims description 27
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims description 26
- 229910052698 phosphorus Inorganic materials 0.000 claims description 26
- 239000011574 phosphorus Substances 0.000 claims description 26
- 125000001931 aliphatic group Chemical group 0.000 claims description 21
- 239000003513 alkali Substances 0.000 claims description 15
- 150000001875 compounds Chemical class 0.000 claims description 13
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims description 12
- 125000003710 aryl alkyl group Chemical group 0.000 claims description 12
- 125000000623 heterocyclic group Chemical group 0.000 claims description 12
- 125000004433 nitrogen atom Chemical group N* 0.000 claims description 11
- MCTWTZJPVLRJOU-UHFFFAOYSA-N 1-methyl-1H-imidazole Chemical compound CN1C=CN=C1 MCTWTZJPVLRJOU-UHFFFAOYSA-N 0.000 claims description 10
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 10
- ITQTTZVARXURQS-UHFFFAOYSA-N 3-methylpyridine Chemical compound CC1=CC=CN=C1 ITQTTZVARXURQS-UHFFFAOYSA-N 0.000 claims description 8
- LSDPWZHWYPCBBB-UHFFFAOYSA-N Methanethiol Chemical compound SC LSDPWZHWYPCBBB-UHFFFAOYSA-N 0.000 claims description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims description 8
- 125000001072 heteroaryl group Chemical group 0.000 claims description 8
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 229910021529 ammonia Inorganic materials 0.000 claims description 5
- 125000001181 organosilyl group Chemical group [SiH3]* 0.000 claims description 5
- 229910052717 sulfur Inorganic materials 0.000 claims description 5
- 230000000694 effects Effects 0.000 claims description 4
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 4
- 125000006239 protecting group Chemical group 0.000 claims description 4
- 125000003903 2-propenyl group Chemical group [H]C([*])([H])C([H])=C([H])[H] 0.000 claims description 3
- 125000003368 amide group Chemical group 0.000 claims description 3
- 125000004475 heteroaralkyl group Chemical group 0.000 claims description 3
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims description 3
- 125000003396 thiol group Chemical group [H]S* 0.000 claims description 3
- XYFCBTPGUUZFHI-UHFFFAOYSA-N Phosphine Chemical compound P XYFCBTPGUUZFHI-UHFFFAOYSA-N 0.000 claims 4
- 229910000073 phosphorus hydride Inorganic materials 0.000 claims 2
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 150000008300 phosphoramidites Chemical class 0.000 abstract description 4
- 238000003786 synthesis reaction Methods 0.000 abstract description 4
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 abstract description 3
- 125000001424 substituent group Chemical group 0.000 abstract description 3
- 150000001298 alcohols Chemical class 0.000 abstract description 2
- 150000003573 thiols Chemical class 0.000 abstract 2
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 27
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 24
- IJGRMHOSHXDMSA-UHFFFAOYSA-N nitrogen Substances N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 16
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- 125000000217 alkyl group Chemical group 0.000 description 11
- 239000002585 base Substances 0.000 description 10
- 238000006243 chemical reaction Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 229920006395 saturated elastomer Polymers 0.000 description 8
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 description 6
- UYTPUPDQBNUYGX-UHFFFAOYSA-N guanine Chemical compound O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 150000003512 tertiary amines Chemical class 0.000 description 6
- RWRDLPDLKQPQOW-UHFFFAOYSA-N tetrahydropyrrole Substances C1CCNC1 RWRDLPDLKQPQOW-UHFFFAOYSA-N 0.000 description 6
- GLUUGHFHXGJENI-UHFFFAOYSA-N Piperazine Chemical compound C1CNCCN1 GLUUGHFHXGJENI-UHFFFAOYSA-N 0.000 description 5
- KDCGOANMDULRCW-UHFFFAOYSA-N Purine Natural products N1=CNC2=NC=NC2=C1 KDCGOANMDULRCW-UHFFFAOYSA-N 0.000 description 5
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical class C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 description 5
- 125000003277 amino group Chemical group 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 239000011782 vitamin Substances 0.000 description 5
- 229940088594 vitamin Drugs 0.000 description 5
- 229930003231 vitamin Natural products 0.000 description 5
- 235000013343 vitamin Nutrition 0.000 description 5
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 description 4
- 239000004215 Carbon black (E152) Substances 0.000 description 4
- YNAVUWVOSKDBBP-UHFFFAOYSA-N Morpholine Chemical compound C1COCCN1 YNAVUWVOSKDBBP-UHFFFAOYSA-N 0.000 description 4
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical compound C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 description 4
- CZPWVGJYEJSRLH-UHFFFAOYSA-N Pyrimidine Chemical compound C1=CN=CN=C1 CZPWVGJYEJSRLH-UHFFFAOYSA-N 0.000 description 4
- SMWDFEZZVXVKRB-UHFFFAOYSA-N Quinoline Chemical compound N1=CC=CC2=CC=CC=C21 SMWDFEZZVXVKRB-UHFFFAOYSA-N 0.000 description 4
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 239000012043 crude product Substances 0.000 description 4
- 229940104302 cytosine Drugs 0.000 description 4
- 229910052736 halogen Inorganic materials 0.000 description 4
- 150000002367 halogens Chemical class 0.000 description 4
- 239000002773 nucleotide Substances 0.000 description 4
- 125000003729 nucleotide group Chemical group 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- 150000003722 vitamin derivatives Chemical class 0.000 description 4
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 description 3
- YXFVVABEGXRONW-UHFFFAOYSA-N Toluene Chemical compound CC1=CC=CC=C1 YXFVVABEGXRONW-UHFFFAOYSA-N 0.000 description 3
- 125000002723 alicyclic group Chemical group 0.000 description 3
- 150000001408 amides Chemical class 0.000 description 3
- 125000001797 benzyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C([H])([H])* 0.000 description 3
- 125000002837 carbocyclic group Chemical group 0.000 description 3
- 125000004432 carbon atom Chemical group C* 0.000 description 3
- 125000004093 cyano group Chemical group *C#N 0.000 description 3
- UAOMVDZJSHZZME-UHFFFAOYSA-N diisopropylamine Chemical compound CC(C)NC(C)C UAOMVDZJSHZZME-UHFFFAOYSA-N 0.000 description 3
- 239000006260 foam Substances 0.000 description 3
- 150000002460 imidazoles Chemical class 0.000 description 3
- 125000006574 non-aromatic ring group Chemical group 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 125000002221 trityl group Chemical group [H]C1=C([H])C([H])=C([H])C([H])=C1C([*])(C1=C(C(=C(C(=C1[H])[H])[H])[H])[H])C1=C([H])C([H])=C([H])C([H])=C1[H] 0.000 description 3
- FNEQHKCQXDKYEO-UHFFFAOYSA-N 1-benzylpyrrole Chemical compound C1=CC=CN1CC1=CC=CC=C1 FNEQHKCQXDKYEO-UHFFFAOYSA-N 0.000 description 2
- HYZJCKYKOHLVJF-UHFFFAOYSA-N 1H-benzimidazole Chemical compound C1=CC=C2NC=NC2=C1 HYZJCKYKOHLVJF-UHFFFAOYSA-N 0.000 description 2
- BAXOFTOLAUCFNW-UHFFFAOYSA-N 1H-indazole Chemical compound C1=CC=C2C=NNC2=C1 BAXOFTOLAUCFNW-UHFFFAOYSA-N 0.000 description 2
- KJUGUADJHNHALS-UHFFFAOYSA-N 1H-tetrazole Chemical compound C=1N=NNN=1 KJUGUADJHNHALS-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- KXDHJXZQYSOELW-UHFFFAOYSA-M Carbamate Chemical compound NC([O-])=O KXDHJXZQYSOELW-UHFFFAOYSA-M 0.000 description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 description 2
- 241000701022 Cytomegalovirus Species 0.000 description 2
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- GHASVSINZRGABV-UHFFFAOYSA-N Fluorouracil Chemical compound FC1=CNC(=O)NC1=O GHASVSINZRGABV-UHFFFAOYSA-N 0.000 description 2
- NYHBQMYGNKIUIF-UUOKFMHZSA-N Guanosine Chemical compound C1=NC=2C(=O)NC(N)=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O NYHBQMYGNKIUIF-UUOKFMHZSA-N 0.000 description 2
- BAVYZALUXZFZLV-UHFFFAOYSA-N Methylamine Chemical compound NC BAVYZALUXZFZLV-UHFFFAOYSA-N 0.000 description 2
- JGFZNNIVVJXRND-UHFFFAOYSA-N N,N-Diisopropylethylamine (DIPEA) Chemical compound CCN(C(C)C)C(C)C JGFZNNIVVJXRND-UHFFFAOYSA-N 0.000 description 2
- PCNDJXKNXGMECE-UHFFFAOYSA-N Phenazine Natural products C1=CC=CC2=NC3=CC=CC=C3N=C21 PCNDJXKNXGMECE-UHFFFAOYSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 239000000908 ammonium hydroxide Substances 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 229910052786 argon Inorganic materials 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 description 2
- 239000000460 chlorine Substances 0.000 description 2
- 229910052801 chlorine Inorganic materials 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 229930182470 glycoside Natural products 0.000 description 2
- 125000001188 haloalkyl group Chemical group 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 2
- 150000002475 indoles Chemical class 0.000 description 2
- AWJUIBRHMBBTKR-UHFFFAOYSA-N isoquinoline Chemical compound C1=NC=CC2=CC=CC=C21 AWJUIBRHMBBTKR-UHFFFAOYSA-N 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000012044 organic layer Substances 0.000 description 2
- WCPAKWJPBJAGKN-UHFFFAOYSA-N oxadiazole Chemical compound C1=CON=N1 WCPAKWJPBJAGKN-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- HGPWSFNGQIFXOV-UHFFFAOYSA-N propan-2-yloxyphosphonamidous acid Chemical compound CC(C)OP(N)O HGPWSFNGQIFXOV-UHFFFAOYSA-N 0.000 description 2
- 150000003217 pyrazoles Chemical class 0.000 description 2
- 150000003233 pyrroles Chemical class 0.000 description 2
- XSCHRSMBECNVNS-UHFFFAOYSA-N quinoxaline Chemical compound N1=CC=NC2=CC=CC=C21 XSCHRSMBECNVNS-UHFFFAOYSA-N 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 239000002342 ribonucleoside Substances 0.000 description 2
- 150000003335 secondary amines Chemical class 0.000 description 2
- 235000017557 sodium bicarbonate Nutrition 0.000 description 2
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- ILMRJRBKQSSXGY-UHFFFAOYSA-N tert-butyl(dimethyl)silicon Chemical group C[Si](C)C(C)(C)C ILMRJRBKQSSXGY-UHFFFAOYSA-N 0.000 description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 description 2
- 125000001544 thienyl group Chemical group 0.000 description 2
- 210000001541 thymus gland Anatomy 0.000 description 2
- 125000003866 trichloromethyl group Chemical group ClC(Cl)(Cl)* 0.000 description 2
- 125000002023 trifluoromethyl group Chemical group FC(F)(F)* 0.000 description 2
- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 125000004169 (C1-C6) alkyl group Chemical group 0.000 description 1
- JYEUMXHLPRZUAT-UHFFFAOYSA-N 1,2,3-triazine Chemical compound C1=CN=NN=C1 JYEUMXHLPRZUAT-UHFFFAOYSA-N 0.000 description 1
- DBNDHLFGQSAGHB-UHFFFAOYSA-N 1,2-oxazole;1,3-thiazole Chemical compound C=1C=NOC=1.C1=CSC=N1 DBNDHLFGQSAGHB-UHFFFAOYSA-N 0.000 description 1
- CFQPVBJOKYSPKG-UHFFFAOYSA-N 1,3-dimethylimidazol-2-one Chemical compound CN1C=CN(C)C1=O CFQPVBJOKYSPKG-UHFFFAOYSA-N 0.000 description 1
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- MNEIJGDSFRHGMS-UHFFFAOYSA-N 1-(phenylmethyl)benzimidazole Chemical compound C1=NC2=CC=CC=C2N1CC1=CC=CC=C1 MNEIJGDSFRHGMS-UHFFFAOYSA-N 0.000 description 1
- 125000004961 1-arylpyrazolyl group Chemical group 0.000 description 1
- FCEHBMOGCRZNNI-UHFFFAOYSA-N 1-benzothiophene Chemical compound C1=CC=C2SC=CC2=C1 FCEHBMOGCRZNNI-UHFFFAOYSA-N 0.000 description 1
- KKKDZZRICRFGSD-UHFFFAOYSA-N 1-benzylimidazole Chemical compound C1=CN=CN1CC1=CC=CC=C1 KKKDZZRICRFGSD-UHFFFAOYSA-N 0.000 description 1
- NJZQOCCEDXRQJM-UHFFFAOYSA-N 1-benzylindole Chemical compound C1=CC2=CC=CC=C2N1CC1=CC=CC=C1 NJZQOCCEDXRQJM-UHFFFAOYSA-N 0.000 description 1
- LQTMKPZRKXYOLG-UHFFFAOYSA-N 1-benzylpurine Chemical compound C1=NC2=NC=NC2=CN1CC1=CC=CC=C1 LQTMKPZRKXYOLG-UHFFFAOYSA-N 0.000 description 1
- AKQAJYLKBCWJBV-UHFFFAOYSA-N 1-benzylpyrazole Chemical compound C1=CC=NN1CC1=CC=CC=C1 AKQAJYLKBCWJBV-UHFFFAOYSA-N 0.000 description 1
- DBMKVSWIHRGBBC-UHFFFAOYSA-N 1-benzylpyrrolo[2,3-b]pyridine Chemical compound C1=CC2=CC=CN=C2N1CC1=CC=CC=C1 DBMKVSWIHRGBBC-UHFFFAOYSA-N 0.000 description 1
- SEULWJSKCVACTH-UHFFFAOYSA-N 1-phenylimidazole Chemical compound C1=NC=CN1C1=CC=CC=C1 SEULWJSKCVACTH-UHFFFAOYSA-N 0.000 description 1
- QUKPALAWEPMWOS-UHFFFAOYSA-N 1h-pyrazolo[3,4-d]pyrimidine Chemical compound C1=NC=C2C=NNC2=N1 QUKPALAWEPMWOS-UHFFFAOYSA-N 0.000 description 1
- FRUWMYWEARDNTC-UHFFFAOYSA-N 2,3,3a,4-tetrahydro-1h-indole Chemical compound C1C=CC=C2NCCC21 FRUWMYWEARDNTC-UHFFFAOYSA-N 0.000 description 1
- MWBWWFOAEOYUST-UHFFFAOYSA-N 2-aminopurine Chemical compound NC1=NC=C2N=CNC2=N1 MWBWWFOAEOYUST-UHFFFAOYSA-N 0.000 description 1
- 125000002103 4,4'-dimethoxytriphenylmethyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)(C1=C([H])C([H])=C(OC([H])([H])[H])C([H])=C1[H])C1=C([H])C([H])=C(OC([H])([H])[H])C([H])=C1[H] 0.000 description 1
- GAMYYCRTACQSBR-UHFFFAOYSA-N 4-azabenzimidazole Chemical compound C1=CC=C2NC=NC2=N1 GAMYYCRTACQSBR-UHFFFAOYSA-N 0.000 description 1
- 108091027075 5S-rRNA precursor Proteins 0.000 description 1
- LOSIULRWFAEMFL-UHFFFAOYSA-N 7-deazaguanine Chemical compound O=C1NC(N)=NC2=C1CC=N2 LOSIULRWFAEMFL-UHFFFAOYSA-N 0.000 description 1
- IKZRFGGARFKJOA-UHFFFAOYSA-N 7h-purin-8-amine Chemical compound C1=NC=C2NC(N)=NC2=N1 IKZRFGGARFKJOA-UHFFFAOYSA-N 0.000 description 1
- JJTNLWSCFYERCK-UHFFFAOYSA-N 7h-pyrrolo[2,3-d]pyrimidine Chemical compound N1=CN=C2NC=CC2=C1 JJTNLWSCFYERCK-UHFFFAOYSA-N 0.000 description 1
- 229960005508 8-azaguanine Drugs 0.000 description 1
- MSSXOMSJDRHRMC-UHFFFAOYSA-N 9H-purine-2,6-diamine Chemical compound NC1=NC(N)=C2NC=NC2=N1 MSSXOMSJDRHRMC-UHFFFAOYSA-N 0.000 description 1
- WKBOTKDWSSQWDR-UHFFFAOYSA-N Bromine atom Chemical compound [Br] WKBOTKDWSSQWDR-UHFFFAOYSA-N 0.000 description 1
- 239000007848 Bronsted acid Substances 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 239000002841 Lewis acid Substances 0.000 description 1
- SECXISVLQFMRJM-UHFFFAOYSA-N N-Methylpyrrolidone Chemical compound CN1CCCC1=O SECXISVLQFMRJM-UHFFFAOYSA-N 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Natural products O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 125000003545 alkoxy group Chemical group 0.000 description 1
- 125000005428 anthryl group Chemical group [H]C1=C([H])C([H])=C2C([H])=C3C(*)=C([H])C([H])=C([H])C3=C([H])C2=C1[H] 0.000 description 1
- 150000008378 aryl ethers Chemical class 0.000 description 1
- 125000004429 atom Chemical group 0.000 description 1
- XYOVOXDWRFGKEX-UHFFFAOYSA-N azepine Chemical compound N1C=CC=CC=C1 XYOVOXDWRFGKEX-UHFFFAOYSA-N 0.000 description 1
- HONIICLYMWZJFZ-UHFFFAOYSA-N azetidine Chemical compound C1CNC1 HONIICLYMWZJFZ-UHFFFAOYSA-N 0.000 description 1
- SIKJAQJRHWYJAI-UHFFFAOYSA-N benzopyrrole Natural products C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 1
- GDTBXPJZTBHREO-UHFFFAOYSA-N bromine Substances BrBr GDTBXPJZTBHREO-UHFFFAOYSA-N 0.000 description 1
- 229910052794 bromium Inorganic materials 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 150000001728 carbonyl compounds Chemical class 0.000 description 1
- 150000003857 carboxamides Chemical class 0.000 description 1
- NEHMKBQYUWJMIP-UHFFFAOYSA-N chloromethane Chemical group ClC NEHMKBQYUWJMIP-UHFFFAOYSA-N 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- HPNMFZURTQLUMO-UHFFFAOYSA-N diethylamine Chemical compound CCNCC HPNMFZURTQLUMO-UHFFFAOYSA-N 0.000 description 1
- 229940043279 diisopropylamine Drugs 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 125000001495 ethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 150000002195 fatty ethers Chemical class 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 229960002949 fluorouracil Drugs 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 150000002240 furans Chemical class 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 125000005842 heteroatom Chemical group 0.000 description 1
- 150000002402 hexoses Chemical class 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- ZLTPDFXIESTBQG-UHFFFAOYSA-N isothiazole Chemical compound C=1C=NSC=1 ZLTPDFXIESTBQG-UHFFFAOYSA-N 0.000 description 1
- 150000007517 lewis acids Chemical class 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000002757 morpholinyl group Chemical group 0.000 description 1
- 125000001624 naphthyl group Chemical group 0.000 description 1
- 125000000449 nitro group Chemical group [O-][N+](*)=O 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 230000000269 nucleophilic effect Effects 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 229940124276 oligodeoxyribonucleotide Drugs 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 125000000636 p-nitrophenyl group Chemical group [H]C1=C([H])C(=C([H])C([H])=C1*)[N+]([O-])=O 0.000 description 1
- 125000004193 piperazinyl group Chemical group 0.000 description 1
- 150000003053 piperidines Chemical class 0.000 description 1
- 125000005936 piperidyl group Chemical group 0.000 description 1
- 230000004853 protein function Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 150000003222 pyridines Chemical class 0.000 description 1
- 125000004076 pyridyl group Chemical group 0.000 description 1
- JWVCLYRUEFBMGU-UHFFFAOYSA-N quinazoline Chemical compound N1=CN=CC2=CC=CC=C21 JWVCLYRUEFBMGU-UHFFFAOYSA-N 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- YBRBMKDOPFTVDT-UHFFFAOYSA-N tert-butylamine Chemical compound CC(C)(C)N YBRBMKDOPFTVDT-UHFFFAOYSA-N 0.000 description 1
- 150000003527 tetrahydropyrans Chemical group 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 125000001113 thiadiazolyl group Chemical group 0.000 description 1
- 125000004568 thiomorpholinyl group Chemical group 0.000 description 1
- 125000005270 trialkylamine group Chemical group 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 125000003774 valeryl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D275/00—Heterocyclic compounds containing 1,2-thiazole or hydrogenated 1,2-thiazole rings
- C07D275/04—Heterocyclic compounds containing 1,2-thiazole or hydrogenated 1,2-thiazole rings condensed with carbocyclic rings or ring systems
- C07D275/06—Heterocyclic compounds containing 1,2-thiazole or hydrogenated 1,2-thiazole rings condensed with carbocyclic rings or ring systems with hetero atoms directly attached to the ring sulfur atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/06—Pyrimidine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
- C07F9/00—Compounds containing elements of Groups 5 or 15 of the Periodic Table
- C07F9/02—Phosphorus compounds
- C07F9/06—Phosphorus compounds without P—C bonds
- C07F9/22—Amides of acids of phosphorus
- C07F9/24—Esteramides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Saccharide Compounds (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
提供了一种在活化剂存在下用亚磷酰化试剂对醇或硫醇进行亚磷酰化作用的方法。活化剂具有式(I)结构：其中p为0或1－4的整数，R在每次出现时为取代基。优选X7为0且p为0。所使用的活化剂通常为与有机碱形成的盐络合物。优选的醇或硫醇包括核苷和低聚核苷酸。该方法特别适用于亚磷酰胺的合成。
Description
本发明涉及对醇或硫醇进行亚磷酰化(phosphitylation)的方法，尤其涉及对核苷进行亚磷酰化而形成核苷亚磷酰胺的方法。
合成的低聚核苷酸是检测基因和病毒疾病的重要诊断工具。而且，低聚核苷酸和修饰的低聚核苷酸作为抑制基因表达或蛋白质功能的治疗候选物质是很有用的。由于FDA已批准将低聚核苷酸类似物用于治疗细胞巨化病毒(CMV)，而且其它几种低聚核苷酸类似物目前也正在进行临床试验，因此大规模合成作为治疗候选物质的低聚核苷酸已变得越来越重要。每个临床试验中都需要数千克的纯化低聚核苷酸类似物。
目前用于制备低聚核苷酸的主要方法是亚磷酰胺法。对于更大量的低聚核苷酸的不断增长的需求也使亚磷酰胺化合物的需求量相应增加。通常，亚磷酰胺化合物是在活化剂的存在下，将核苷用亚磷酰化试剂进行亚磷酰化作用来制备。最常使用的活化剂为亲核活化剂1H-四唑。但是，1H-四唑为爆炸性物质，因此用在大规模合成中可能带来危险。
为了更容易地制备诊断和治疗用途的低聚核苷酸，则需要可促进亚磷酰化作用又不会增加副产物的非爆炸性活化剂。
根据本发明，其提供了在活化剂存在下，用亚磷酰化试剂对醇或硫醇进行亚磷酰化作用的方法，其特征在于，活化剂具有式1的结构：
式1中，p为0或1-4的整数。R在每次出现时为取代基，优选各自独立为卤素、取代或未取代的脂肪族基团、-NR1R2、-OR3、-OC(O)R3、-C(O)OR3、氰基、取代或未取代的芳基、取代或未取代的杂环基、-CHO、-COR3、-NHCOR3、取代或未取代的芳烷基、卤代烷基(如，三氟甲基和三氯甲基)，或-SR3。优选，R为卤素、取代或未取代的脂肪族基团、-NR1R2、-OR3、-OC(O)R3、-C(O)OR3或氰基。或者，两个相邻R基团与其所连接的碳原子一起形成饱和或不饱和六元环，优选芳环。R1和R2各自独立为-H、取代或未取代的脂肪族基团、取代或未取代的芳基、取代或未取代的芳烷基；或与其所连接的氮原子一起形成杂环基。R为取代或未取代的脂肪族基团、取代或未取代的芳基、取代或未取代的芳烷基。X为O或S。优选，X为O。特别优选X为O，且p为0。
优选所使用的式1化合物为与有机碱形成的盐络合物。
在许多实施方案中，该醇或硫醇为含有游离羟基或硫醇基团的核苷或低聚核苷酸，而且该醇或硫醇还包括含有天然核苷戊糖和非天然核苷糖，如己糖的核苷和低聚核苷酸。当醇或硫醇为核苷或低聚核苷酸时，其通常为被保护的脱氧核糖核苷、被保护的核糖核苷、被保护的寡脱氧核糖核苷酸、被保护的寡核糖核苷酸，或具有脱氧核糖核苷酸和核糖核苷酸部分的被保护的低聚核苷酸，其中各个部分都包括游离的3’-或5’，优选3’-羟基或硫醇基团，且最优选为3’-羟基基团。
可按照本发明的方法进行亚磷酰化的醇和硫醇包括式2的化合物：
其中，A代表
其中，X1每次出现时独立为-O-或-S-。优选，X1每次出现时均为-O-。X2每次出现时独立为-O-、-S-、-CH2或-(CH2)2-。优选，X2每次出现时均为-O-。X3每次出现时独立为O或S。在更优选的实施方案中，X1和X2每次出现时各自为-O-。R4为醇保护基团或硫醇保护基团。优选，R为对酸不稳定的保护基团。R5每次出现时独立为-H、-F、-OR6、-NR7R8、-SR9，或取代或未取代的脂肪族基团，例如甲基或烯丙基。R10每次出现时独立为磷保护基团，其通常为合成低聚核苷酸时使用的的可裂分的磷保护基团，并优选为取代或未取代的脂肪族基团、取代或未取代的芳基或取代或未取代的芳烷基，例如式-CH2CH2CN、-CH2CH2CN、-CH2CH2-Si(CH3)2C6H5、-CH2CH2-S(O)2-CH2CH3、-CH2CH2-C6H4-NO2、-CH2CH2-Si(CH3)2C6H5、-CH2CH2-S(O)2-CH2CH3或-CH2CH2-C6H4-NO2的基团。R6每次出现时独立为-H、取代或未取代的脂肪族基团(如，甲基、乙基、甲氧基乙基或烯丙基)、取代或未取代的芳基、取代或未取代的芳烷基、醇保护基团、特别是碱-不稳定的保护基团，或甲硅烷基保护基团，或-(CH2)q-NR11R12。R7和R8每次出现时各自独立为-H、取代或未取代的脂肪族基团，或氨保护基团。或者，R7和R8与其所连接的氮原子一起为杂环基。R9每次出现时独立为-H、取代或未取代的脂肪族基团，或硫醇保护基团。R11和R12各自独立为-H、取代或未取代的芳基、取代或未取代的杂芳基、取代或未取代的脂肪族基团、取代或未取代的芳烷基、取代或未取代的杂芳烷基，或氨基保护基团。或者，R11和R12与其所连接的氮原子一起形成杂环基。q为1-约6的整数。s为0或正整数。优选，s为0、1或2，最优选为0。B各自独立为-H、天然或非天然核碱、被保护的核碱、被保护的天然或非天然核碱、杂环或被保护的杂环。
核苷碱包括天然存在的碱，例如腺嘌呤、鸟嘌呤、胞嘧啶、胸腺嘧啶和尿嘧啶，还包括经修饰的碱，例如7-脱氮鸟嘌呤、7-脱氮-8-氮鸟嘌呤、5-丙炔基胞嘧啶、5-丙炔基尿嘧啶、7-脱氮腺嘌呤、7-脱氮-8-氮腺嘌呤、7--脱氮-6-氧嘌呤、6-氧嘌呤、3-脱氮腺苷、2-氧-5-甲基嘧啶、2-氧-4-甲硫基-5-甲基嘧啶、2-硫代羰基-4-氧-5-甲基嘧啶、4-氧-5-甲基嘧啶、2-氨基嘌呤、5-氟尿嘧啶、2，6-二氨基嘌呤、8-氨基嘌呤、4-三唑-5-甲基胸腺嘧啶、4-三唑-5-甲基尿嘧啶和次黄嘌呤。
被保护的核苷碱是指其中的反应性碱官能团被保护起来的核苷碱。类似地，被保护的杂环是杂环中的反应性取代基被保护起来的杂环。通常，核苷碱或杂环具有可被氨基保护基团(如酰胺或氨基甲酸酯)保护起来的氨基基团。例如，腺嘌呤和胞嘧啶的氨基基团通常被苯甲酰保护基团所保护，鸟嘌呤中的氨基基团通常被异丁酰基、4-异丙基苯氧乙酰基或叔丁基苯氧乙酰基保护。但是，还可使用其它保护方案，例如甲脒。例如，为了快速解保护，腺嘌呤和鸟嘌呤的氨基基团用苯氧乙酰基进行保护，胞嘧啶的氨基基团用异丁酰基或乙酰基进行保护。除去核碱或杂环保护基团的反应条件要取决于所用的保护基团。使用酰胺保护基团时，可用碱溶液处理低聚核苷酸来除去保护基团，该碱溶液例如为浓缩氢氧化铵溶液、正甲胺溶液或氢氧化铵中的叔丁胺溶液。
氨、羟基和硫醇保护基团对本领域熟练技术人员来说是已知的。氨保护基团的例子参见Greene等，Protective Groups in OrganicSynthesis(1991)，John Wiley & Sons，Inc.，309-405页，该文献所教导的全部内容引入本文作为参考。优选，将氨保护为酰胺或氨基甲酸酯。羟基保护基团的例子参见同上，10-142页，其全部教导引入本文作为参考。可使用的保护基团的例子包括甲硅烷基，特别是三烷基，例如三(C1-4烷基)甲硅烷基。优选的甲硅烷基保护基团为叔丁基二甲基甲硅烷基。优选的羟基保护基团为叔丁基二甲基甲硅烷基。硫醇保护基团的例子参见同上，277-308页，其全部教导引入本文作为参考。
酸不稳定的保护基团是指通过使该基团与Bronsted或Lewis酸接触可将其除去的保护基团。酸不稳定的保护基团对本领域熟练技术人员来说是已知的。常用的酸不稳定保护基团的例子包括取代或未取代的三苯甲基(同上，60-62页)、取代或未取代的四氢吡喃基团(同上，31-34页)、取代或未取代的四氢呋喃基团(同上，36-37页)或pixyl基团(同上，65页)。通常用给电子取代基，例如烷氧基基团取代三苯甲基基团。优选的酸不稳定保护基团为取代或未取代的三苯甲基，例如4，4’-二甲氧基三苯甲基(下文中为“DMT”)。
碱不稳定的保护基团是指通过使该基团与Bronsted或Lewis碱接触可将其除去的保护基团。碱不稳定的保护基团对本领域熟练技术人员来说是已知的。常用的碱不稳定保护基团的例子包括羰基化合物，例如乙酰基、苯甲酰基和新戊酰基。
应当认识到，如果式2表达为给定醇的天然核苷构型(D-异构体)，则本发明同样还可用于相应的合成或非天然醇构型(L-异构体)，以及上述两种构型的混合物中。
可用于本发明方法中的亚磷酰化试剂通常具有下述化学通式：
R13-X6-PX4X5其中，R13表示磷保护基团，通常为合成低聚核苷酸中所用的可裂解的磷保护基团，例如取代或未取代的脂肪族基团或芳烷基，例如甲基、-CH2CH2-Si(CH3)2C6H5、-CH2CH2-S(O)2-CH2CH3、-CH2CH2-C6H4-NO2，并优选为式-CH2CH2CN的基团；取代或未取代的芳香族基团，例如苯基或取代的苯基，例如4-氯苯基、2-氯苯基、2-硝基苯基或4-硝基苯基；X6代表O或S，并优选O；X4和X5可相同或不同，代表离去基团，例如卤素，通常为溴或氯，或-NR14R15，其中R14和R15各自独立代表烷基，优选C1-6烷基基团，或R14和R15与其所连接的N一起形成5-7元环。通常X4和X5中的至少一个基团为式-NR14R15的基团。最优选，X4和X5相同，并且尤其优选X4和X5都为-N[CH(CH3)2]2基团。特别优选X6为0且R15为-CH2CH2CN。
本发明的方法尤其适用于制备亚磷酰胺，尤其是核苷和低聚核苷酸亚磷酰胺。
优选的亚磷酰化试剂的例子包括O-β-氰基乙基-N，N，N’，N’-四异丙基亚磷酰二胺(O-β-cyanoethy1-N，N，N’，N’-tetraisopropylphosphorodiamidite)，(通常称作“tetraphos”)、O-β-氰基乙基-N，N，N’，N’-四甲基亚磷酰二胺、O-β-氰基乙基-N，N，N’，N’-四乙基亚磷酰二胺、双(N，N-二异丙基氨基)-2-甲基三氟乙酰基氨基乙氧基膦、双(N，N-二异丙基氨基)-2-二苯甲基甲硅烷基乙氧基膦和O-β-氰基乙基-双(N-吗啉代)亚磷酰二胺。
本发明的方法通常在0℃-约50℃的温度下进行，优选在环境温度，例如约15℃-约30℃下进行。
有利的是使用基本无水的反应条件。
本发明的许多实施方案是在惰性气氛，例如氮气或氩气下进行。
根据本发明的方法可有利地用于制备核苷亚磷酰胺。因此，本发明的优选方面包括制备下式化合物的方法：
其包括：使下式化合物：
其中，R4如上述定义，并优选为二甲氧基三苯甲基，R5如上述定义；
与下式化合物：
NCCH2CH2O-P(N(R16)2)其中，R16代表C1-6烷基，优选为异丙基；
在活化剂存在下进行反应，其中活化剂包括下式化合物：
及有机碱。
在许多实施方案中，活化剂按照与醇的化学计量摩尔比或小于化学计量摩尔比的量使用，活化剂与醇的摩尔比为约0.4∶1-1∶1，尤其优选为约0.5∶1-0.75∶1。
通常，亚磷酰化试剂按照与醇的化学计量摩尔比或过量使用，亚磷酰化试剂与醇的摩尔比为约1∶1-3∶1，尤其优选为约1∶1-1.5∶1。
在有机碱存在下，本发明中所用的活化剂具有良好的溶解性，尤其是在亚磷酰化反应常用的有机溶剂中。活化剂和有机碱的浓度可达到活化剂在所用溶剂中的溶解度。在优选实施方案中，活化剂和有机碱的存在浓度为约0.01M-约2M，例如约0.05M-约0.5M。通常活化剂和有机碱的存在浓度可高达0.25M，例如约0.1M-约0.25M。在更优选的实施方案中，活化剂和有机碱的摩尔浓度相同。在某些实施方案中，有机溶剂为氯碳化合物，例如二氯甲烷。在优选的实施方案中，有机溶剂包括乙腈。另一个优选实施方案中，有机溶剂包括有机酰胺，例如二甲基甲酰胺、1-甲基-2-吡咯烷酮或1，3-二甲基-2-咪唑啉酮。
有机碱是指pH为7时趋向于接受质子的有机化合物。优选的有机碱为仲胺、叔胺或氮杂环化合物，每种化合物均可被一个或多个取代基取代或未取代。非质子有机碱是指接受质子之前其化学结构中不具有氢键合质子的有机碱。优选将非质子有机碱，例如叔胺和非质子氮杂环化合物与本文中描述的式1化合物结合使用，以改善亚磷酰化反应。
本文中描述的氮杂环化合物包括芳环上具有一个或多个氮原子的杂芳基，和非芳环体系上具有至少一个氮原子的杂脂环族基。优选，氮杂芳基化合物具有5-或6-元芳环，且芳环上具有1-3个氮原子。优选氮杂脂环基化合物为5-或6-元环，通常在环上包括一个或两个氮原子。优选的氮杂环化合物为有机碱。有机碱氮杂环化合物的例子包括嘧啶、1-烷基吡唑，特别是1-(C1-4烷基)吡唑、1-芳基吡唑、1-苄基吡唑、吡嗪、N-烷基嘌呤，特别是N-(C1-4烷基)嘌呤、N-芳基嘌呤、N-苄基嘌呤、N-烷基吡咯，特别是N-(C1-4烷基)吡咯、N-芳基吡咯、N-苄基吡咯、吡啶、N-烷基咪唑，特别是N-(C1-4烷基)咪唑、N-芳基咪唑，特别是N-苯基咪唑、N-苄基咪唑、喹啉、异喹啉、喹喔啉、喹唑啉、N-烷基吲哚，特别是N-(C1-4烷基)吲哚、N-芳基吲哚，N-苄基吲哚、N-烷基苯并咪唑，特别是N-(C1-4烷基)苯并咪唑、N-芳基苯并咪唑、N-苄基苯并咪唑、三嗪、噻唑、1-烷基-7-吖吲哚，特别是1-(C1-4烷基)-7-吖吲哚、1-芳基-7-吖吲哚、1-苄基-7-吖吲哚、吡咯烷、吗啉、哌啶和哌嗪。特别优选的氮杂环化合物为吡啶类，例如吡啶和3-甲基吡啶，和N-(C1-4烷基)咪唑，例如N-甲基咪唑。
叔胺是指一种具有氮原子的有机碱，且其中的氮原子键合于三个碳原子上，往往键合于三个芳基上，通常为苯基，和/或烷基，通常键合于三个烷基上，该烷基包括，例如三烷基胺，例如三甲胺、三乙胺和二异丙基乙基胺。此外，叔胺还可为氮杂环基团，且其中的氮原子为非质子性。氮杂环基团类的叔胺为优选的。氮杂环叔胺的例子为N-烷基吡咯烷、N-芳基吡咯烷、N-烷基吡咯、N-芳基吡咯、N-烷基吗啉、N-芳基吗啉、N-烷基哌啶、N-芳基哌啶、N，N-二烷基哌嗪、N，N-二芳基哌嗪、N-烷基-N-芳基哌嗪、奎宁环、1，5-二氮杂二环[4.3.0]壬-5-烯和1，8-二氮杂二环[5.4.0]十一碳-7-烯。叔胺还可为氮杂芳基化合物或氮杂杂脂环基化合物。
仲胺是指氮键合到单个氢和两个碳原子上的有机碱。通常，氮原子键合到两个烷基或芳基上，或形成氮杂环基团的一部分。仲胺化合物的例子包括二乙胺和二异丙基胺。
特别优选的有机碱包括吡啶、3-甲基吡啶和N-甲基咪唑。
脂肪族基团、芳基、芳烷基、杂芳基、氮杂芳基和杂脂环族基的适当取代基包括芳基、卤代芳基、烷基、卤代烷基(如，三氟甲基和三氯甲基)、脂肪族醚、芳香族醚、苄基、取代的苄基、卤素，尤其是氯和氟、氰基、硝基、-S-(脂肪族或取代的脂肪族基团)，和-S-(芳香族或取代的芳香族基团)。
本文中所用的脂肪族基团包括直链或支链的、完全饱和或含有一个或多个非共轭双键的C1-C18烃，或完全饱和或含有一个或多个非共轭双键的环状C3-C18烃。烷基基团为完全饱和的直链或支链C1-C8烃或C3-C8环状烃。脂肪族基团优选为烷基。
芳基基团包括碳环芳环体系(如，苯基)，和稠合于一个或多个碳环芳烃的碳环芳环体系(如，萘基和蒽基)，或稠合于一个或多个非芳环的芳环体系(如，1，2，3，4-四氢萘基)。
本文中所用的杂环基团包括杂芳基和杂脂环族基。本文中所用的杂芳基包括芳环上具有一个或多个选自硫、氮或氧的杂原子的芳环体系。优选，杂芳基为具有1-4个杂原子的5或6元环体系。本文中所用的脂肪族杂环基为非芳环体系，其优选具有5-6个原子并包括至少一个选自氮、氧和硫的杂原子。杂环基团的例子包括吗啉基、哌啶基、哌嗪基、硫代吗啉基、吡咯烷基、噻唑烷基、四氢thienyl、吖丁啶基、四氢呋喃基、二氧己环基和二氧杂烷基(dioxepanyl)、噻吩基、吡啶基、噻二唑基、噁二唑基、吲哚基、呋喃、吡咯、咪唑、吡唑、三唑、嘧啶、吡嗪、噻唑、异噁唑、异噻唑、四唑、噁二唑、苯并噻吩、苯并咪唑、吲哚、四氢吲哚、吖吲哚、吲唑、喹啉、咪唑并吡啶、嘌呤、吡咯并[2，3-d]嘧啶和吡唑并[3，4-d]嘧啶。
本文中所用的芳烷基基团是指通过烷基进行连接的芳香族取代基。优选的芳烷基包括苄基。
本文中所用的杂芳烷基基团是指通过烷基进行连接的杂芳基取代基。
通过下面实施例对本发明进行非限制性的说明。
实施例1
按照下列步骤制备糖精的N-甲基咪唑盐。将糖精悬浮在乙腈中，并向悬浮液中滴加相对于糖精为1.1当量的N-甲基咪唑。减压浓缩反应混合物以形成结晶盐，用醚或己烷洗涤该结晶盐以除去痕量的N-甲基咪唑和乙腈。
实施例2
使用O-β-氰基乙基-N，N，N’，N’-四异丙基亚磷酰胺，和作为活化剂的糖精N-甲基咪唑盐来对一系列核苷进行亚磷酰化作用。
一般方法：
向适当尺寸的烧瓶中加入核苷(1.5mmol)，并与20mL吡啶一起共沸蒸馏两次(旋转蒸发器)以干燥固体。用Ar吹扫烧瓶并向烧瓶中加入15mL乙腈。室温下搅拌混合物直至获得澄清溶液。向混合物中加入O-β-氰基乙基-N，N，N’，N’-四异丙基亚磷酰胺(Tetraphos)，之后加入糖精的N-甲基咪唑盐。室温下搅拌混合物，同时用HPLC监测反应直至结束。反应结束时，用30mL乙酸乙酯稀释混合物，并用2×25mL的碳酸氢钠饱和水溶液和25mL氯化钠饱和水溶液洗涤有机混合物。分离有机层并用MgSO4干燥。过滤悬浮液，并使用旋转蒸发器除去溶剂。剩余物经真空干燥得到泡沫状物。
表1：amidite合成结果
| Tctraphos(当量) | 活化剂(当量) | 核苷 | Rxn时间(h) | ％amiditea(HPLC) | 产率b(％) |
| 1.2 | 0.6 | 5’-DMT-N-Bz-deoxyA | 7 | 91.9 | 84 |
| 1.2 | 0.5 | 5’-DMT-N-Bz-deoxyA | 5 | 91.5 | 86 |
| 1.2 | 1.0 | 5’-DMT-N-Bz-deoxyA | 5 | 89.8 | 87 |
| 1.2 | 0.5 | 5’-DMT-N-iBu-deoxyG | 16 | 79.1 | 85 |
| 1.2 | 0.5 | 5’-DMT-N-Ac-2’-OMeC | 55 | 89.3 | 85 |
| 1.2 | 0.6 | 5’-DMT-N-Bz-deoxyC | 8 | 91.1 | 79 |
| 1.2 | 0.6 | 5’-DMT-2’-TBDMS-L-U | 16 | 82.b | 82 |
| 1.2 | 0.6 | 5’DMT-Ni-Bu-2’-TBDMS-L-G | 1b | 59.3 | 84 |
| 2.2 | 0.6 | 5’-DMT-N-iBu-2’-TBDMS-L-G | 1b | 87.4 | 82 |
a％amidite＝粗产物中的％amidite
b产率＝粗产物产率
实施例2
向500mL圆底烧瓶中加入5’-DMT-N-Bz-2’-去氧腺苷(18.00g，27.37mmol)，并通过加入和蒸发(旋转蒸发器)2×200mL的甲苯来共沸干燥固体。剩余物真空干燥16h。在氩气氛下将剩余物溶解在乙腈(180mL)中，并加入O-β-氰基乙基-N，N，N’，N’-四异丙基亚磷酰二胺(9.90g，32.84mmol)。搅拌混合物5分钟并加入固体糖精N-甲基咪唑盐(3.63g，13.69mmol)。室温下搅拌混合物，同时用HPLC监测反应的进行。18h后，未观察到进一步反应。向反应混合物中加入乙酸乙酯(200mL)，并用碳酸氢钠饱和水溶液(2×150mL)和氯化钠饱和水溶液(150mL)洗涤有机溶液。分离有机层并用MgSO4干燥。过滤悬浮液，并使用旋转蒸发器除去溶剂。剩余物经真空干燥16h得到白色泡沫状物。
粗产率：23.70g
HPLC：92.5％
使用硅胶(230g)柱对粗产物(23.70g)进行色谱分离。用30％乙酸乙酯/含0.5％三乙胺的己烷装载色谱柱。用2倍柱体积的30％乙酸乙酯/己烷洗涤色谱柱。将粗产物装载在色谱柱上，并用2倍柱体积的30％乙酸乙酯/己烷、2倍柱体积的40％乙酸乙酯/己烷、2倍柱体积的50％乙酸乙酯/己烷和最后的3倍柱体积的70％乙酸乙酯/己烷洗脱色谱柱。当TLC(8∶3的乙酸乙酯∶己烷)检测出产物时收集馏分。将含有所希望的产物的馏分合并起来，并使用旋转蒸发器除去馏分中的溶剂。真空干燥剩余物16h得到白色泡沫状物。
产率：17.55g(75％)
HPLC：97.5％
31P NMR：99.3％
Claims (13)
1、一种在活化剂存在下，用亚磷酰化试剂对醇或硫醇进行亚磷酰化的方法，其特征在于，活化剂具有式1的结构：
其中，p为0或1-4的整数，R在每次出现时为取代基，X7为O或S。
2、根据权利要求1的方法，其中X7为O，且p为0。
3、根据权利要求1或2的方法，其中使用式1化合物与有机碱形成的盐络合物。
4、根据权利要求3的方法，其中有机碱选自吡啶、3-甲基吡啶和N-甲基咪唑。
5、根据前述任一权利要求的方法，其中醇或硫醇为包括游离羟基或硫醇基团的核苷或低聚核苷酸。
6、根据权利要求5的方法，其中对包括游离3’-羟基基团的核苷进行亚磷酰化作用。
7、根据前述任一权利要求的方法，其中亚磷酰化试剂具有下列化学通式：
R13-X6-PX4X5
其中，R13表示磷保护基团，X6代表O或S，X4和X5可相同或不同，代表离去基团。
8、根据权利要求7的方法，其中R13表示取代或未取代的脂肪族基团或芳烷基，或取代或未取代的芳香族基团，X6为O且X4和X5各自独立代表-NR14R15，其中R14和R15各自独立代表C1-6烷基基团，或R14和R15与其所连接的N一起形成5-7元环。
9、根据权利要求8的方法，其中亚磷酰化试剂选自O-β-氰基乙基-N，N，N’，N’-四异丙基亚磷酰二胺、O-β-氰基乙基-N，N，N’，N’-四甲基亚磷酰二胺、O-β-氰基乙基-N，N，N’，N’-四乙基亚磷酰二胺、双(N，N-二异丙基氨基)-2-甲基三氟乙酰基氨基乙氧基膦、双(N，N-二异丙基氨基)-2-二苯甲基甲硅烷基乙氧基膦和O-β-氰基乙基-双(N-吗啉代)亚磷酰二胺。
10、一种制备下式化合物的方法：
其包括：使下式化合物：
与下式化合物：
NCCH2CH2O-P(N(R16)2)2
在活化剂存在下进行反应，其中活化剂包括下式化合物：
及有机碱，其中，R4为醇保护基团，R5为-H、-F、-OR6、-NR7R8、-SR9，或取代或未取代的脂肪族基团，例如甲基或烯丙基，R6每次出现时独立为-H、取代或未取代的脂肪族基团、取代或未取代的芳基、取代或未取代的芳烷基、醇保护基团、或-(CH2)q-NR11R12，R7和R8各自独立为-H、取代或未取代的脂肪族基团，或氨保护基团，或者R7和R8与其所连接的氮原子一起为杂环基，R9为-H、取代或未取代的脂肪族基团，或硫醇保护基团，R11和R12各自独立为-H、取代或未取代的芳基、取代或未取代的杂芳基、取代或未取代的脂肪族基团、取代或未取代的芳烷基、取代或未取代的杂芳烷基，或氨基保护基团，或者R11和R12与其所连接的氮原子一起形成杂环基，q为1-约6的整数，B为-H、天然或非天然核碱、被保护的核碱、被保护的天然或非天然核碱、杂环或被保护的杂环，R16代表C1-6烷基，优选异丙基。
11、根据权利要求10的方法，其中有机碱选自吡啶、3-甲基吡啶和N-甲基咪唑。
12、根据权利要求10或11的方法，其中R5为H、OMe或OCH2CH2OMe。
13、根据权利要求10或11的方法，其中R4为酸不稳定的保护基团，且R5为OR6，其中R6为碱不稳定的保护基团或甲硅烷基保护基团。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US41818502P | 2002-10-15 | 2002-10-15 | |
| US60/418,185 | 2002-10-15 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1705675A true CN1705675A (zh) | 2005-12-07 |
| CN1329408C CN1329408C (zh) | 2007-08-01 |
Family
ID=32107901
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2003801014035A Expired - Lifetime CN1329408C (zh) | 2002-10-15 | 2003-10-08 | 亚磷酰化方法 |
Country Status (10)
| Country | Link |
|---|---|
| US (1) | US7247720B2 (zh) |
| EP (1) | EP1554300B1 (zh) |
| JP (1) | JP4756863B2 (zh) |
| KR (1) | KR101065715B1 (zh) |
| CN (1) | CN1329408C (zh) |
| AT (1) | ATE362937T1 (zh) |
| AU (1) | AU2003269239A1 (zh) |
| CA (1) | CA2501565C (zh) |
| DE (1) | DE60313993T2 (zh) |
| WO (1) | WO2004035599A1 (zh) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AR036122A1 (es) * | 2001-07-03 | 2004-08-11 | Avecia Biotechnology Inc | Un complejo de sal que comprende un n-alquilimidazol y una 1,1-dioxo-1,2-dihidro-1l6-benzo [d]-isotiazol-3-ona y un metodo para sintetizar oligonucleotidos utilizando la quimica de fosforamidita |
| GB0229443D0 (en) * | 2002-12-18 | 2003-01-22 | Avecia Ltd | Process |
| KR20220123300A (ko) * | 2020-01-08 | 2022-09-06 | 닛토덴코 가부시키가이샤 | 분절-타입 아미디트(segment-type amidite)를 사용한 핵산 합성 방법 |
| WO2023140040A1 (ja) * | 2022-01-21 | 2023-07-27 | 国立大学法人東海国立大学機構 | アミダイトモノマー |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5034534A (en) * | 1988-07-15 | 1991-07-23 | E. I. Du Pont De Nemours And Company | Process for producing saccharin, saccharin analogues or their salts |
| US5662111A (en) * | 1991-01-28 | 1997-09-02 | Cosman; Eric R. | Process of stereotactic optical navigation |
| US6034534A (en) * | 1995-05-25 | 2000-03-07 | Kiyota; Shigeo | Laminated contact probe for inspection of ultra-microscopic pitch |
| US6274725B1 (en) * | 1998-06-02 | 2001-08-14 | Isis Pharmaceuticals, Inc. | Activators for oligonucleotide synthesis |
| AR036122A1 (es) * | 2001-07-03 | 2004-08-11 | Avecia Biotechnology Inc | Un complejo de sal que comprende un n-alquilimidazol y una 1,1-dioxo-1,2-dihidro-1l6-benzo [d]-isotiazol-3-ona y un metodo para sintetizar oligonucleotidos utilizando la quimica de fosforamidita |
-
2003
- 2003-10-08 CA CA2501565A patent/CA2501565C/en not_active Expired - Lifetime
- 2003-10-08 KR KR1020057006226A patent/KR101065715B1/ko active IP Right Grant
- 2003-10-08 DE DE60313993T patent/DE60313993T2/de not_active Expired - Lifetime
- 2003-10-08 JP JP2004544427A patent/JP4756863B2/ja not_active Expired - Lifetime
- 2003-10-08 WO PCT/GB2003/004312 patent/WO2004035599A1/en active IP Right Grant
- 2003-10-08 AT AT03751018T patent/ATE362937T1/de not_active IP Right Cessation
- 2003-10-08 EP EP03751018A patent/EP1554300B1/en not_active Expired - Lifetime
- 2003-10-08 US US10/531,323 patent/US7247720B2/en not_active Expired - Lifetime
- 2003-10-08 AU AU2003269239A patent/AU2003269239A1/en not_active Abandoned
- 2003-10-08 CN CNB2003801014035A patent/CN1329408C/zh not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| CA2501565C (en) | 2011-03-15 |
| EP1554300A1 (en) | 2005-07-20 |
| KR20050055004A (ko) | 2005-06-10 |
| WO2004035599A1 (en) | 2004-04-29 |
| CA2501565A1 (en) | 2004-04-29 |
| CN1329408C (zh) | 2007-08-01 |
| JP2006508081A (ja) | 2006-03-09 |
| DE60313993T2 (de) | 2008-01-24 |
| EP1554300B1 (en) | 2007-05-23 |
| US7247720B2 (en) | 2007-07-24 |
| AU2003269239A1 (en) | 2004-05-04 |
| KR101065715B1 (ko) | 2011-09-19 |
| DE60313993D1 (de) | 2007-07-05 |
| US20060069247A1 (en) | 2006-03-30 |
| JP4756863B2 (ja) | 2011-08-24 |
| ATE362937T1 (de) | 2007-06-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5512668A (en) | Solid phase oligonucleotide synthesis using phospholane intermediates | |
| US5883237A (en) | Oligonucleotides having Rp and Sp linkages at predetermined locations | |
| CN1066456C (zh) | 具有2'－醚基的寡核苷酸 | |
| US5646267A (en) | Method of making oligonucleotides and oligonucleotide analogs using phospholanes and enantiomerically resolved phospholane analogues | |
| HUT64555A (en) | A method for linking nucleosides with syloxane bridge | |
| CN1479745A (zh) | 硫代磷酸寡核苷酸的制备方法 | |
| EP1119578B1 (en) | Improved process for oligonucleotide synthesis | |
| US5955600A (en) | Method for the synthesis of nucleotide or oligonucleotide phosphoramidites | |
| CN1329408C (zh) | 亚磷酰化方法 | |
| CN110891961B (zh) | 制备伊美司他(imetelstat)的改良方法 | |
| JP4824931B2 (ja) | オリゴヌクレオチド・シントン類を精製する方法 | |
| US20220275019A1 (en) | Hydrocinnamoyl protected riboguanosine phosphoramidites for decreasing depyrimidination from alkyl amine exposure during final deprotection | |
| US7872121B2 (en) | Process for the removal of exocyclic base protecting groups | |
| US6441150B1 (en) | Compounds for the Synthesis of Nucleotide or oligonucleotide phosphoramidites | |
| US6103891A (en) | Method for the synthesis of nucleotide or oligonucleotide phosphoramidites | |
| CA1214735A (en) | Nucleotide compound preparation and method for producing the same | |
| EP4089099A1 (en) | Nucleic acid synthesis method using segment-type amidite | |
| Stell | Synthesis of Phosphonoacetate RNA and a Two-Step RNA Synthesis | |
| Stell | Synthesis of phosphonoacetate RNA and a two-step RNA synthesis approach | |
| JP2006077013A (ja) | Ｒｎａ合成に有用な水酸基の新規保護基を有する化合物 | |
| WO2003045969A1 (en) | Coupling reagent for h-phosphonate chemistry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: NITTO DENKO ALWAYS CO., LTD. Free format text: FORMER NAME: AVECIA BIOTECHNOLOGY INC. |
|
| CP01 | Change in the name or title of a patent holder |
Address after: Massachusetts, USA Patentee after: GIRINDUS AMERICA, Inc. Address before: Massachusetts, USA Patentee before: Avecia Biotechnology Inc. |
|
| CX01 | Expiry of patent term | ||
| CX01 | Expiry of patent term |
Granted publication date: 20070801 |