Patent EA-002603-B1 (EA)
Title: Method and reagents for the quantification of solid-phase reactions using fluorine nmr
Inventor: NAGULAPALLI VASANT K
IPC: G01N24/08
Filed: 1998-12-14
Abstract: 1. A method of quantitating a solid-phase reaction comprising: (a) reacting a solid-phase reaction component or a fluorine-containing solid-phase reaction component with a reactant or fluorine-containing reactant to form a fluorine-containing solid-phase reaction product; (b) obtaining a <19>F NMR spectrum of the fluorine-containing solid-phase reaction product; and (c) comparing the integral corresponding to the fluorine-containing solid-phase reaction product <19>F resonance to the integral corresponding to a standard <19>F resonance. 2. The method according to Claim 1 wherein the <19>F NMR spectra are obtained using magic angle spinning. 3. The method according to Claim 1 wherein the internal standard is a fluorine-containing solid-phase reaction component of the formula   wherein    is a solid support optionally containing one or more fluorine atoms; L is absent or a linking group optionally containing one or more fluorine atoms, provided that at least one of the components selected from solid support and the linking group contains at least one fluorine atom; and B is a functional group suitable for reaction with a reactant to form a fluorine-containing solid-phase reaction product. 4. The method according to Claim 3 wherein the solid support contains one or more fluorine atoms. 5. The method according to Claim 3 wherein   is a solid support; L is a group of formula   A is absent or is selected from   -C(O)-, -YC(O)-, -SO2-, -NR<7>SO2-, -CHR<7>-, -CHR<7>Y- and -CHR<7>YC(O)(CH2)m-; or when B is halogen, NHP, OW or SO2Z; D is CH or N; P is H or an amine protecting group; W is H, NHP, NPR<9>, NC(O)Cl, C(O)R<9>, C(O)NR<10>R<11>, C(O)OR<9>, SO2R<9> or C(O)-imidazol-1-yl; Y is -O- or-NR<8>-; Z is Cl, OH, ORa or NR<9>R<12>; R<1> is F, or when one of R<16>, R<17>, R<18> and R<19> is F, R<1> is H, alkyl, alkoxy, halogen, CN or NO2; R<2>, R<3> and R<4> are independently H, alkyl, alkoxy, halogen, CN or NO2, or one of R<2>, R and R<4>, taken together with one of R<5> and R<6> and the carbon atoms to which they are attached, define a group of formula   R<5> and R<6> are independently -H, alkyl, phenyl or phenyl substituted with one or more substituents selected from alkyl, alkoxy, halogen, nitrile and -NO2; R<7> and R<8> are independently H or alkyl comprising 1 to 4 carbon atoms; R<9> and R<13> are independently aliphatic or aromatic; R<10> and R<11> are independently H, aliphatic or aromatic; R<12> is -CH2R<13>; R<14>, R<15>, R<16>, R<17>, R<18>, R<19>, R<20>, R<21>, R<22> and R<23> are independently selected from H, alkyl, alkoxy, halogen, -CN and -NO2; m is 0 or 1; n is 1-6; and p is 0, 1 or 2. 6. The method according to Claim 5 wherein R<1>, R<2>, R<3> and R<4> are F; and one of R<5> and R<6> is H and the other of R<5> and R<6> is H or 2,4-dimethoxyphenyl. 7. The method according to Claim 6 wherein B is F, OW or SO2Z. 8. The method according to Claim 7 wherein A is phenylene, -C(O)-, -YC(O)-, -SO2-, -NR<7>SO2- or -CHR<7>O-. 9. The method according to Claim 5 wherein the fluorine-containing solid-phase reaction component is selected from 4-carboxy-2,3,5,6-tetrafluorophenoxymethyl-copoly(styrene-1% divinylbenzene) resin, 4-(O-methylhydroxylamine)-2,3,5,6-tetrafluorophenoxymethyl-copoly(styrene-1% divinylbenzene) resin, 4-(2',4'-dimethoxyphenyl-O-methylhydroxylamine)-2,3,5,6-tetrafluorophenoxymethyl-copoly(styrene-1%-divinylbenzene) resin, 4-hydroxy-2,3,5,6-tetrafluorobenzamidomethyl-polystyrene resin, 2,3,5,6-tetrafluorobenzamidomethyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorobenzamidomethyl-4-sulfonylchloride-polystyrene resin, 4-hydroxy-2,3,5,6-tetrafluorobenzoyloxymethyl-polystyrene resin, 2,3,5,6-tetrafluorobenzoyloxymethyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorobenzoyloxymethyl-4-sulfonylchloride-polystyrene resin, 4-hydroxy-2,3,5,6-pentafluorobenzoyl-polystyrene resin, 2,3,5,6-tetrafluorobenzoyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorobenzoyl-4-sulfonylchloride-polystyrene resin, 4-hydroxy-2,3,5,6-tetrafluorophenylsulfonamidomethyl-polystyrene resin, 2,3,5,6-tetrafluorophenylsulfonamidomethyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorophenylsulfonamidomethyl-4-sulfonylchloride-polystyrene resin, N-(4-hydroxy-2,3,5,6-tetrafluorobenzoyl)-piperidinomethyl-polystyrene resin, N-(2,3,5,6-tetrafluorobenzoyl-4-sulfonic acid)-piperidinomethyl-polystyrene resin, N-(2,3,5,6-tetrafluorobenzoyl-4-sulfonyl chloride)-piperidinomethyl-4-polystyrene resin, N-(4-hydroxy-2,3 ,5 ,6-tetrafluorophenylsulfonyl)-piperidinomethyl-polystyrene resin, N-((2,3,5,6-tetrafluorophenyl-4-sulfonic acid)sulfonyl)-piperidinomethyl-polystyrene resin, N-((2,3,5,6-tetrafluorophenyl-4-sulfonylchloride)sulfonyl)-piperidinomethyl-polystyrene resin, 4-hydroxy-2,3,5,6-tetrafluorophenyl-polystyrene resin, 2,3,5,6-tetrafluorophenyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorophenyl-4-sulfonylchloride polystyrene resin, 4-hydroxy-2,3,5,6-tetrafluorophenylsulfonyl-polystyrene resin, 2,3,5,6-tetrafluorophenylsulfonyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorophenylsulfonyl-4-sulfonylchloride-polystyrene resin. 10. The method according to Claim 5 wherein the fluorine-containing solid-phase reaction component is selected from 4-carboxy-2,3,5,6-tetrafluorophenoxymethyl-copoly(styrene-1% divinylbenzene) resin, 4-(O-methylhydroxylamine)-2,3,5,6-tetrafluorophenoxymethyl-copoly(styrene-1% divinylbenzene) resin, 4-(2',4'-dimethoxyphenyl-O-methylhydroxylamine)-2,3,5,6-tetra-fluorophenoxymethyl-copoly(styrene-1% divinylbenzene) resin, 4-hydroxy-2,3,5,6-tetrafluorobenzamidomethyl-polystyrene resin, 2,3,5,6-tetrafluorobenzamidomethyl-4-sulfonic acid-polystyrene resin, 2,3,5,6-tetrafluorobenzamidomethyl-4-sulfonylchloride-polystyrene resin, 4-hydroxy-2,3,5,6-pentafluorobenzoyl-polystyrene resin, 2,3,5,6-tetrafluorobenzoyl-4-(sulfonic acid)-polystyrene resin, 2,3,5,6-tetrafluorobenzoyl-4-sulfonylchloride-polystyrene resin, 4-hydroxy-2,3,5,6-tetrafluorophenylsulfonamidomethyl-polystyrene resin, 2,3,5,6-tetrafluorophenylsulfonamidomethyl-4-(sulfonic acid)-polystyrene resin and 2,3,5,6-tetrafluorophenylsulfonamidomethyl-4-sulfonylchloride-polystyrene resin. 11. The method according to Claim 5 wherein the fluorine-containing solid-phase reaction component is 4-hydroxy-2,3,5,6-tetrafluorobenzamidomethyl-polystyrene resin. 12. The method according to Claim 1 wherein the solid-phase reaction component is chloromethylated polystyrene resin and the fluorine-containing reactant is 4-fluorophenol. 13. A fluorine-containing solid-phase reaction component of formula   wherein   is a solid support containing one or more fluorine atoms; LG is absent or a linking group wherein LG is optionally substituted by one or more fluorine atoms; and B is a functional group suitable for reaction with a reactant to form a fluorine-containing solid-phase reaction product. 14. The fluorine-containing solid-phase reaction component according to Claim 13 which is prepared by polymerizing a mixture comprising at least one fluorine-containing monomer. 15. The fluorine-containing solid-phase reaction component according to Claim 14 wherein the fluorine-containing monomer is 4-fluorostyrene. 16. The fluorine-containing solid-phase reaction component according to Claim 14 which is prepared by polymerizing a mixture of 4-fluorostyrene, 1,4-divinylbenzene and 4-vinylbenzylchloride.