Azacyanine dyes and use thereof

A fluorescent dye of formula A

• or a salt thereof, wherein
• Z is selected from the group consisting of NR17 and +NR17R18;
• Q is independently H or selected from the groups a), b), and c) consisting of:
• a) Halide selected from Cl, Br, I;
• R19U, —OR19U, —SR19U and —NR19R20U, wherein R19 is a single bond;
• or wherein R19 and R20 may independently be an optical properties modifying group, and are independently selected from the group consisting of:
• H, linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic C5-, C6- or C7 aromatic ring which can be substituted by a linear or branched C1-C6 alkyl group;
• and homocyclic and heterocyclic 5-, 6- and 7-membered aromatic rings which can be substituted by a linear or branched C1-C6 alkyl group, wherein preferably one of R19 and R20 is not aromatic in case of —NR19R20U;
• —(CH2O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and U is a physiochemistry modifying group selected from the group consisting of:
• —(CH2)mSO3−, —(CH2)mC(O)O−, —(CH2)mP(O)O22−, —(CH2)mNH2;
• —(CH2)mNHR32;
• —(CH2)mNR32R33 wherein m is an integer from 0 to 6, and wherein R32 and R33 are independently of each other an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl;
• and wherein in case of —(CH2)mNH2;
• —(CH2)mNHR32;
• —(CH2)mNR32R33 the N atom may be bond to a further substituent R34 to form a quaternary N atom, and wherein R34 is in all the above cases independently selected from H, and an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl,
• b) R21L, —OR21L, —SR21L and —NR21R22L wherein R21 and R22 may independently be an optical properties modifying group and are independently selected from the group consisting of:
• H, linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic rings which can be substituted by a linear or branched C1-C6 alkyl group, wherein preferably one of R21 and R22 is not aromatic in case of —NR21R22;
• —(CH2—O—CH2)xCH2-L wherein x is an integer from 1 to 50;
• and L is a linker which can form a covalent bond with a targeting agent,
• c) R19, —OR19, —SR19 and —NR19R20 wherein R19 and R20 wherein R19 and R20 may independently be an optical properties modifying group and are independently selected from the group consisting of:
• H, linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic ring which can be substituted by a linear or branched C1-C6 alkyl group;
• and homocyclic and heterocyclic 5-, 6- and 7-membered aromatic rings which can be substituted by a linear of branched C1-C6 alkyl group, wherein preferably one of R19 and R20 is not aromatic in case of —NR19R20;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• or wherein R19 and R20, together with the N atom to which they are attached, form a 5- or 6-membered heterocycle optionally containing one further heteroatom selected from O and N, wherein the heterocycle can be substituted by a linear or branched, cyclic or non cyclic C1-C6 alkyl group, in particular 4-cyclohexylpiperazinyl;
• L is selected from the group consisting of:
• —NH2, —OH, —SH, —C(O)O−, —C(O)Cl, —(CO)O(CO)R27, —C(O)NHNH2, —C(O)—C(O)OR28, wherein R27 is selected from the group consisting of H, alkyl and aryl;
• wherein R28 is derived from substituted and unsubstituted N-hydroxysuccinimide, substituted and unsubstituted N-hydroxysulfosuccinimide, nitrophenol, fluorophenol each bound via —O—;
• azide N3−, —NCO, —NCS, —CHO, —COCH2I, phosphoramidityl, phthalamidyl, maleimide, an alkyne group in particular —C═CR31 wherein R31 is H or a C1-C8 alkyl group, sulfonate esters, alkyl halides, acyl halides, propargylglycine, a pentanoyl group, in particular pentanoyl chloride, pentynoic acid, propargylic acid, 6-aminobenzo[d]thiazole-2-carbonitrile, 6-hydroxybenzo[d]thiazole-2-carbonitrile, a 1,2-aminothiol group, in particular L-cysteine or D-cysteine;
• R1 and R2 are absent, H or independently selected from the group:
• a) linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- and 7-membered aromatic rings which can be substituted by a linear of branched C1-C6 alkyl group;
• and —(CH2—O—CH2)CH2— wherein x is an integer from 1 to 50,
• b) R23L wherein R23 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic C5-, C6- or C7-aryl group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- and 7-membered aromatic rings which can be substituted by a linear or branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and L is a linker which can form a covalent bond with a targeting agent,
• c) R23U, wherein R23 is a single bond;
• or wherein R23 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic rings which can be substituted by a linear or branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and U is a physiochemistry modifying group selected from the group consisting of:
• —(CH2)mSO3−, —((C2)mC(O)−, —(CH2)mP(O)O22−—(CH2)mNR2—(CH2)mNH2;
• —(CH2)mNHR32;
• (CH2)mNR32R33 wherein m is an integer from 0 to 6, and wherein R32, R33 are independently of each other an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl, wherein m in an integer from 0 to 6;
• R17 and R20 are independently H or selected from the group consisting of:
• a) linear and branched, non-cyclic or cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- and 7-membered aromatic rings which can be substituted by a linear or branched C1-C6 alkyl group, wherein preferably one of R17 and R18 is not aromatic;
• and —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50,
• b) R24L wherein R24 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group, wherein preferably one of R17 and R20 is not aromatic;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and L is a linker which can form a covalent bond with a targeting agent,
• c) R24U, wherein R24 is a single bond;
• or wherein R24 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group, wherein preferably one of R17 and R18 is not aromatic;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and U is a physiochemistry modifying group selected from the group consisting of:
• —(CH2)mSO3−, —(CH2)mC(O)O—(CH2)mNH2;
• —(CH2)mP(O)O22−—(CH2)mNR2—(CH2)mNH2;
• —(CH2)mNHR32;
• (CH2)mNR32R33 wherein m is am integer from 0 to 6, and wherein R32, R33, are independently of each other an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl, wherein m is an integer from 0 to 6;
• A6, A7, A8, A9, and A10, A11, A12, A13 are C, N or +N, and either
• A)
• form a 6-membered aromatic ring which together with the pyrrolin derived ring to which they are attached form an indol or an azaindol system, which indol system can comprise a total of 1 N atoms, and which azaindol system can comprise a total of 2 N atoms;
• R3, R4, R5, R6, R7, R8 R9 R10 are independently H or selected from the group consisting of:
• a) halide selected from Cl, Br, I;
• R25H and OR25H, wherein R25 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50,
• b) R25L and OR25L, wherein R23 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2—wherein x is an integer from 1 to 50;
• and L is a linker which can form a covalent bond with a targeting agent,
• c) R25U and OR25U wherein R25 is a single bond;
• or wherein R25 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear or branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and U is a physiochemistry modifying group selected from the group consisting of:
• —(CH2)mSO3−, —(CH2)mC(O)O−, —(CH2)mP(O)O22−—(CH2)mNR2—(CH2)mNH2;
• —(CH2)mNHR32;
• (CH2)mNR32R33 wherein m is an integer from 0 to 6, and wherein R32, R33, are independently of each other an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl, wherein m in an integer from 0 to 6;
• and wherein OR25H, OR25L and OR25U are present only when O is attached to a C atom;
• Or
• B)
• A6, A7, A8, A9, and A10, A11, A12, A13 are C, N, or +N and form a 6-membered aromatic ring which together with the pyrrolin derived ring to which they are attached form an indol or an azaindol system, and to which indol or azaindol system a further 6-membered ring is annulated which is formed by at least two of the substitutents R3, R4, R5, R6, or R7, R8 R9 R10, resulting in a trinuclear ring in which 1, 2 or 3 C atoms may be replaced by N or +N and which are substituted by R11, R12, R13, R14, and R15, R16, R17;
• R18;
• R11, R12, R13, R14, and R15, R16, R17, R18 are independently H or selected from the group consisting of:
• a) halide selected from Cl, Br, I;
• R26H and OR26H, wherein R26 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50,
• b) R26L and OR26L, wherein R26 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic C5-, C6- or C7-aryl group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic C5-, C6- or C7-aromatic groups which can be substituted by a linear or branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and L is a linker which can form a covalent bond with a targeting agent,
• c) R26U and OR26U, wherein R26 is a single bond;
• or wherein R26 is selected from the group:
• linear and branched, non-cyclic and cyclic, substituted and unsubstituted C1-20 alkyl, wherein the said alkyl group can be single or multiple substituted by a homocyclic or heterocyclic 5-, 6- or 7-membered aromatic group which can be substituted by a linear or branched C1-C6 alkyl group;
• homocyclic and heterocyclic 5-, 6- or 7-membered aromatic groups which can be substituted by a linear of branched C1-C6 alkyl group;
• —(CH2—O—CH2)xCH2— wherein x is an integer from 1 to 50;
• and U is a physiochemistry modifying group selected from the group consisting of:
• —(CH2)mSO3−, —(CH2)mC(O)O−, and
• d) —(CH2)mP(O)O22−—(CH2)mNR2—(CH2)mNH2;
• —(CH2)mNHR32;
• (CH2)mNR32R33 wherein m is an integer from 0 to 6, and wherein R32, R33, are independently of each other an alkyl group having from 1-12, preferably 1-8, more preferably 1-4 C atoms, in particular methyl or ethyl;
• X and Y are selected from the group consisting of CR29R30, where R29 and R30 are each independently selected from H, unsubstituted and substituted linear or branched, cyclic or non-cyclic C1-C6 alkyl,
• E and E′ are independently selected from H, unsubstituted and substituted linear or branched, cyclic or non-cyclic C1-C6 alkyl.