Novel fluorescent dyes, labeled conjugates and analytical methods

A dye compound represented by the formula

• wherein
• D is selected from —C—R5, —C-(A)q-R6, E+(G−), and
• X is selected from oxygen, sulfur, selenium, N—B, and —C(═O)O—;
• B is selected from hydrogen, alkyl having from 1 to 10 carbon atoms, aryl, heteroaryl, acyl, aroyl having from 1 to 10 carbon atoms and heteroaroyl having from 1 to 10 carbon atoms);
• Z is carbon or nitrogen;
• Za is selected from carbon, nitrogen and —C-(A)q-R6 ;
• Zb is selected from carbon, N-alkyl ad N-aryl;
• A is a divalent linking group having alternating single and multiple bonds;
• E+is oxygen or N+—R7, where R7 is hydrogen, cyano, alkyl, linear or branched, preferably having from 1 to 10 carbon atoms, arylalkyl, preferably having from 1 to 10 carbon atoms or heteroarylalkyl, preferably having from 1 to 10 carbon atoms;
• (G−) is a counterion;
• Y is a proton transfer group;
• Ar is an aromatic or a heteroaromatic moiety;
• PG is a protective group;
• R1, R2 and R3 each independently is selected from hydrogen, an electron withdrawing group and an electron releasing group;
• or
• R1 and R2 taken together with the carbon atoms to which they are attached represent the carbon atoms necessary to form a substituted or unsubstituted unsaturated carbocycic or heteroaromatic ring;
• R4 is selected from hydrogen, an electron withdrawing group and an electron releasing group;
• R5 is selected from hydrogen and a 5 or 6 member aromatic or heteroaromatic;
• or
• R2 and R5 taken together with the carbon atoms to which they are attached represent the carbon atoms necessary to form an unsaturated carbocyclic or heteroaromatic ring;
• R6 is
• Where R11 is H, alkyl, aryl, heteroaryl, H2W or R13R14;
• R12 is hydrogen, alkyl, aryl or heteroaryl;
• R13 is hydrogen;
• R11 and R12 taken together with the nitrogen and oxygen atoms to which they are attached represent the carbon atoms necessary to form a substituted or unsubstituted, saturated or unsaturated ring as shown in the examples below.
• R14 is alkyl, aryl or heteroaryl;
• W is an anion;
• n is an integer of from 0 to 4;
• and
• q is 0 or 1
• provided that at least Za or D is —C-(A)q-R6.