Dyes for analysis of protein aggregation
AssigneeEnzo Biochem, Inc.
InventorsWayne Forrest Patton, Sergiy M. Yarmoluk, Praveen Pande, Vladyslava Kovalska, Lijun Dai, Kateryna Volkova, Jack Coleman, Mykhaylo Losytskyy, Anthony Ludlam, Anatoliy Balanda, Dee Shen
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Abstract
Provided are dyes and compositions which are useful in a number of applications, such as the detection and monitoring protein aggregation, kinetic studies of protein aggregation, neurofibrillary plaques analysis, evaluation of protein formulation stability, and analysis of molecular chaperone activity.
Claims21
A compound comprising the structure
- wherein m and n are independently 1, 2 or 3;
- wherein L is a linker arm comprising carbon, sulfur, oxygen, nitrogen, or any combination thereof;
- wherein R1, R2, R3, R4, R9, R10, R11, R12, R13, R14, R15, R16, R19, R20, R21 and R22 are independently hydrogen, halogen, amino, ammonium, nitro, sulfo, sulfonamide, carboxy, ester, cyano, phenyl, benzyl, an alkyl group wherein the alkyl group is saturated or unsaturated, linear or branched, substituted or unsubstituted, an alkoxy group wherein the alkoxy group is saturated or unsaturated, branched or linear, substituted or unsubstituted, or when taken in combination R1 and R2, or R3 and R4 or R9 and R10, or R11 and R12, or R13 and R14, or R15 and R16, or R19 and R20, or R21 and R22 form a five or six membered ring wherein the ring is saturated or unsaturated, substituted or unsubstituted, and wherein R9 and R10, or R11 and R12, or R13 and R14, or R15 and R16 can comprise alkyl chains that are joined together, wherein a quinoline moiety can be formed;
- wherein R7, R8, R17 and R18 are independently hydrogen, Z, an alkyl group wherein the alkyl group is saturated or unsaturated, linear or branched, substituted or unsubstituted, an alkoxy group wherein the alkoxy group is saturated or unsaturated, branched or linear, substituted or unsubstituted, or when taken together, R7 and R8 and R17 and R18, may form a 5 or 6 membered ring wherein the ring is saturated or unsaturated, substituted or unsubstituted;
- wherein Z comprises a carboxyl group (CO2−), a carbonate ester (COER25), a sulfonate (SO3−), a sulfonate ester (SO2ER25), a sulfoxide (SO2R25), a sulfone (SO2CR25R26R27), a sulfonamide (SO2NR25R26), a phosphate (PO4═) a phosphate monoester (PO3−ER25), a phosphate diester (PO2ER25ER26), a phosphonate (PO3═) a phosphonate monoester (PO2−ER25) a phosphonate diester (POER25ER26), a thiophosphate (PSO3═), a thiophosphate monoester (PSO2−ER25) a thiophosphate diester (PSOER25ER26), a thiophosphonate (PSO2═), a thiophosphonate monoester (PSO−ER25) a thiophosphonate diester (PSER25ER26), a phosphonamide (PONR25R26NR28R29), its thioanalogue (PSNR25R26NR28R29), a phosphoramide (PONR25R26NR27NR28R29), its thioanalogue (PSNR25R26NR27NR28R29), a phosphoramidite (PO2R25NR28R29) or its thioanalogue (POSR25NR28R29) where E can be independently O or S;
- wherein R25, R26, R27, R28, and R29 are independently a hydrogen, an unsubstituted straight-chain, branched or cyclic alkyl, alkenyl or alkynyl group, a substituted straight-chain, branched or cyclic alkyl, alkenyl or alkynyl group wherein one or more C, CH or CH2 groups are substituted with an O atom, N atom, S atom, or NH group, or an unsubstituted or substituted aromatic group;
- wherein Z is attached directly, or indirectly through a second linker arm comprising carbon, sulfur, oxygen, nitrogen, and any combinations thereof and wherein the second linker arm may be saturated or unsaturated, linear or branched, substituted or unsubstituted or any combinations thereof;
- and
- wherein R5, R6, R23 and R24 can independently be hydrogen or an alkyl group wherein the alkyl group is saturated or unsaturated, linear or branched, substituted or unsubstituted, or when taken in combination R5 and R6 or R2 and R5 or R3 and R6 or R23 and R24 or R22 and R23 or R20 and R24 form a five or six membered ring wherein the ring is saturated or unsaturated, substituted or unsubstituted.
A compound exhibiting at least three times increased fluorescence in the presence of an aggregated form of a protein when compared to the fluorescence exhibited when the compound is in the presence of the unaggregated form of the protein, wherein the compound is D95, D97, L-30, L-33, Lu-1, Lu-2, S-8, S13. S22, S25, S33, S39, S42, S43, S48, S49, SL2131, SL2592, Tio-1, TOL-2, TOL-3, TOL-5, TOL-6, TOL-7, TOL-11, YA-1, YA-3, YAT2134, YAT2135, YAT2148, YAT2149, YAT2150, YAT2213, YAT2214 or YAT2324.
A multi-dye composition comprising at least three dyes, wherein each of the at least three dyes exhibits increased fluorescence in the presence of an aggregated form of a protein when compared to the fluorescence exhibited when the compound is in the presence of the unaggregated form of the protein.
21-115. (canceled)
Dependencies are inferred from the claim text.