5(6)-methyl substituted fluorescein derivatives

Claim 2

Fluorescein derivatives of the following formulas: ##STR47## wherein R is selected from the group consisting of hydroxy and protected hydroxy; and wherein X is selected from the group consisting of hydroxy, protected hydroxy, amino, protected amino, thiol, protected thiol, a leaving group, a Michael acceptor, phosphoramidite, phosphonate and a linking group.

Claim 3

The fluorescein derivative according to claim 1 wherein the protected hydroxyl groups are selected from the group consisting of alkyl esters, aryl ethers, silyl ethers, esters, carbonates, and carbamates.

Claim 4

The fluorescein derivative according to claim 1 wherein the protected amino groups are selected from the group consisting of carbamates, amides, cyclic imides, N-benzyl derivatives, imine derivatives, silyl derivatives, and N-sulfonyl derivatives.

Claim 5

The fluorescein derivative according to claim 1 wherein the thiol groups are selected from the group consisting of thioethers, thioesters, thiocarbonates, thiocarbamates and disulfides.

Claim 6

The fluorescein derivative according to claim 1 wherein the Michael acceptor is of the formula --O--"MA" or --N(H) n "MA", wherein "MA" is the Michael acceptor of the formula: ##STR48## wherein a, b and c independently can be hydrogen, alkyl, and aryl and wherein U is chosen from --CH(═O), --C(═O)R 1 , --C(═O)NH 2 , --CN, --NO 2 , --S(═O)R 1 --S(═O) 2 R 1 , and n is 0 or 1.

Claim 7

The fluorescein derivative according to claim 5 wherein said ##STR49## Michael acceptor is wherein when X is a linking group it is selected from the group consisting of the formula --O--A--B and --N(H) n --A--B, wherein n=0 or 1 to satisfy the nitrogen valency; wherein A is a spacer group consisting of from 0 to 50 carbon atoms and heteroatoms, including not more than ten heteroatoms, arranged in a straight or branched chain, saturated or unsaturated, with the provisos that: (a) not more than two heteroatoms may be directly linked in the sequence --O--A--B or --N(H) n --A--B, (b) the sequence --O--A--B or --N(H) n --A--B cannot contain --O--O--, and (c) in the sequence --N(H) n --A--B, when n=0, then N--A taken together may comprise a ring of not more that 6 atoms; and (d) that branchings may occur only on carbon atoms; and wherein B is a reactive functional group selected from --C(═O)OH, --NH 2 , --CH(═O), halo (--Cl, --Br, --I), sulfonate ester(OS(═O) 2 -alkyl, --OS(≡0 O) 2 -aryl), Michael acceptor, phosphoramidite, and phosphonate.