Amyloid binding nitrogen-linked compounds for the antemortem diagnosis of alzheimer's disease, in vivo imaging and prevention of amyloid deposits

An in vivo method for detecting amyloid deposits in a subject, comprising the steps of:

• (a) administering a detectable quantity of radiolabeled Chrysamine G or a Chrysamine G derivative wherein R1-R7 or R10-R49 are radiolabeled and are selected from the group consisting of 131I, 123I, 76Br, 75Br, 18F, CH2—CH21318F, O—CH2—CH2—18F, CH2—CH2—CH21318F, O—CH2—CH2—CH2—18F, 19F, 125I and a 11C or 13C labeled moeity selected from the group consisting of a lower alkyl group, (CH2)nOR′, CF3, CH2—CH2—F, O—CH2—CH2—CH2—F, CH2—CH2—CH2—F, O—CH2—CH2—CH2—F, CN, (C═O)—R′, (C═O)N(R′)2, O(CO)R′, OR′, SR′, N(R′)2, COOR′, Rph, CR′═CR′—Rph, and CR′2—CR′2Rph=, wherein n, R′ and Rph are as defined below, and
• (b) detecting the binding of said compound or a salt thereof to amyloid deposit in said subject;
• wherein said Chrysamine G derivative is an amyloid binding compound of Formula I or a water soluble toxic salt thereof:
• wherein:
• is selected from the group consisting of N═N—Q, CR′═N—Q, N═CR′—Q, CR′2—NR′—Q, NR′—CR′2—Q, (CO)—NR′—Q, NR′—(CO)—Q and NR′—NR′—Q (where R′ independently represents H or a lower alkyl group);
• X is C(R″)2,
• (wherein each R″ independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′ where n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R″), a tetrazole or oxadiazole of the form:
• wherein R′ is H or a lower alkyl group);
• or X is CR′═CR′, N═N, C═O, O, NR′, (where R′ represents H or a lower alkyl group), S, or SO2;
• each R1 and R2 independently is selected from the group consisting of H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, a tri-alkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R1 and R2), tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group), or a triazene of the form:
• (wherein R8 and R9 are lower alkyl groups) or
• and at least one of R2 is not H, OCH3, CH3 or halogen;
• each Q is independently selected from one of the following structures, each of which contains a carboxylic acid or an acid like functionality selected from the group consisting of hydroxy, sulfhydryl, tetrazole, oxadiazole, isoindol-1,3 (2H)-dione, benzisoxazole, 2,3-benzodiazine-1,4 (2H,3H)-dione, 2,3-benzoxazine-1,4 (3H)-dione, (2H) 1,3-benzoxazine-2,4 (3H)-dione, (3H) 2-benzazine-1,3 (2H)-dione, and NO2:
• IA, IB, IC, ID, IE, IF, and IG, wherein
• IA has the following structure:
• wherein:
• each of R3, R4, R5, R6, or R7 is independently defined the same as R1 above and wherein at least one of R3, R4, R5, R6, or R7 is a hydroxy, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• IB has the following structure:
• wherein:
• each of R10, R11, R12, R13, R14, R15, or R16 independently is defined the same as R1 above and, wherein at least one of R10, R11, R12, R13, R14, R15, or R16 is a hydroxy, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• and wherein at least one of R1 or R2 is a halogen when the compound of Formula I is a 4,4′-diazobiphenyl compound;
• IC has the following structure:
• wherein:
• each of R17, R18, R19, R20, or R21 independently is defined the same as R1 above;
• ID has the following structure:
• wherein each of R22, R23, or R24 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IE has the following structure:
• wherein:
• each of R25, R26, or R27 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IF has the following structure:
• wherein:
• exactly one of R28, R29, R30, R31, or R32 is the
• of the
• link defined for Formula I above and each other R28, R29, R30, R31, or R32 independently is defined the same as R1 above, and wherein at least one of R28, R29, R30, R31, or R32 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• IG has the following structure:
• wherein:
• exactly one of R33, R34, R35, R36, R37, R38 or R39 is the
• of the
• link defined for Formula I above and each other R33, R34, R35, R36, R37, R38 or R39 independently is defined the same as R1 above and wherein at least one of R33, R34, R35, R36, R37, R38 or R39 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• or wherein:
• is NR′—N═Q (wherein R′ represents H or a lower alkyl group) and each
• is independently selected from one of the following structures:
• IIA or IIB, wherein:
• IIA has the following structure
• wherein
• each of R40-R47 independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SH, COOR′, a tri-alkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tri-alkyl tin, a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group);
• IIB has the following structure:
• wherein:
• each of R48 and R49 independently is a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, CH2—CH2—CH2F, CN, (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, (C═O)—CH2)n—(C6H5) where n=1, 2, or 3, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group,
• wherein at least one of R48 or R49 is (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, CN, or a tetrazole or oxadiazole of the form:
• (wherein R′ is H or lower alkyl group) in both Q's.

A method of inhibiting cell degeneration and toxicity associated with fibril formation in an amyloidosis-associated condition, said method comprising the step of administering to a subject having or suspected of having such condition a pharmaceutically effective amount of Chrysamine G or a derivative thereof, wherein said derivative of Chrysamine G is an amyloid binding compound of Formula I or a water soluble, non-toxic salt thereof

• wherein:
• is selected from the group consisting of N═N—Q, CR′═N—Q, N═CR′—Q, CR′2—NR′—Q, NR′—CR′2—Q, (CO)—NR′—Q, NR′—(CO)—Q and NR′—NR′—Q (where R′ independently represents H or a lower alkyl group);
• X is C(R″)2,
• (wherein each R″ independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′ where n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2—F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R″), a tetrazole or oxadiazole of the form:
• wherein R′ is H or a lower alkyl group);
• or X is CR′═CR′, N═N, C═O, O, NR′, (where R′ represents H or a lower alkyl group), S, or SO2;
• each R1 and R2 independently is selected from the group consisting of H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, a tri-alkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R1 and R2), a tetrazole or a oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group), or a triazene of the form:
• (wherein R8 and R9 are lower alkyl groups) or
• and at least one of R2 is not H, OCH3, CH3, or halogen;
• each Q is independently selected from one of the following structures, each of which contains a carboxylic acid or an acid like functionality selected from the group consisting of hydroxy, sulfhydryl, tetrazole, oxadiazole, isoindol-1,3 (2H)-dione, 3-hydroxy-1,2-benzisoxazole, 2,3-benzodiazine-1,4 (2H,3H)-dione, 2,3-benzoxiaine-1,4 (3H)-dione, (2H) 1,3-benzoxazine-2,4 (3H)-dione, (3H) 2-benzazine-1,3 (2H)-dione, and NO2:
• IA, IB, IC, ID, IE, IF, and IG, wherein
• IA has the following structure:
• wherein:
• each of R3, R4, R5, R6, or R7 independently is defined the same as R1 above and, wherein at least one of R3, R4, R5, R6, or R7 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's, and wherein at least one of R1 or R2 is a halogen when the compound of Formula I is a 4,4′-diazobiphenyl compound;
• IB has the following structure:
• wherein:
• each of R10, R11, R12, R13, R14, R15, or R16 independently is defined the same as R1 above above and, wherein at least one of R10, R11, R12, R13, R14, R15, or R16 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's, and wherein at least one of R1, or R2 is a halogen when the compound of Formula I is a 4,4′-diazobiphenyl compound;
• IC has the following structure:
• wherein:
• each of R17, R18, R19, R20, or R21, independently is defined the same as R1 above;
• ID has the following structure:
• wherein each of R22, R23, or R24 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IE has the following structure:
• wherein:
• each of R25, R26, or R27 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IF has the following structure:
• wherein:
• exactly one of R28, R29, R30, R31, or R32 is the
• of the
• link defined for Formula I above and each other R28, R29, R30, R31, or R32 independently is defined the same as R1 above, and wherein at least one of R28, R29, R30, R31, or R32 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• IG has the following structure:
• wherein:
• exactly one of R33, R34, R35, R36, R37, R38 or R39 is the
• of the
• link defined for Formula I above and each other R33, R34, R35, R36, R37, R38 or R39 independently is defined the same as R1 above and wherein at least one of R33, R34, R35, R36, R37, R38 or R39 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• or wherein:
• is NR′—N═Q (wherein R′ represents H or a lower alkyl group) and each
• is independently selected from one of the following structures:
• IIA or IIB, wherein:
• IIA has the following structure
• wherein:
• each of R40-R47 independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, a tri-alkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tri-alkyl tin, a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group);
• IIB has the following structure:
• wherein:
• each of R48 and R49 independently is a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, CH2—CH2—CH2F, CN, (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, (C═O)—CH2)n—(C6H5) where n=1, 2, or 3, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group),
• wherein at least one of R48 or R49 is (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, CN, or a tetrazole or oxadiazole of the form:
• (wherein R′ is H or lower alkyl group) in both Q's.

A method of quantifying the amount of amyloid in biopsy or post-mortem tissue comprising the steps of:

• a) incubating radiolabeled Chrysamine G or a derivative of Chrysamine G with a homogenate of biopsy or post-mortem tissue, wherein R1-R7 or R10-R49 are radiolabeled and are selected from the group consisting of 131I, 123I, 76Br, 75Br, 18F, CH2—CH2—18F, O—CH2—CH2—18F, CH2—CH2—CH2—18F, O—CH2—CH2—CH2—18F, 19F, 125I and a 11C or 13C labeled moeity selected from the group consisting of a lower alkyl group, (CH2)nOR′, CF3, CH2—CH2—F, O—CH2—CH2—F, CH2—CH2—CH2—F, O—CH2—CH2—CH2—F, CN, (C═O)—R′, (C═O)N(R′)2, O(CO)R′, OR′, SR′, N(R′)2, COOR′, Rph, CR′═CR′—Rph, and CR′2—CR′2Rph=, wherein n, R′ and Rph are as defined below,
• b) separating the tissue-bound from the tissue-unbound radiolabeled Chrysamine G or Chrysamine G derivative,
• c) quantifying the tissue-bound radiolabeled Chrysamine G or Chrysamine G derivative, and
• d) converting the units of tissue-bound radiolabeled Chrysamine G or Chrysamine G derivative to units of micrograms of amyloid per 100 mg of tissue by comparison with a standard
• wherein said Chrysamine G derivative is an amyloid binding compounds of Formula I or a water soluble toxic salt thereof:
• wherein:
• is selected from the group consisting of N═N—Q, CR′═N—Q, N═CR′—Q, CR′2—NR′—Q, NR′—CR′2—Q, (CO)—NR′—Q, NR′—(CO)—Q and NR′—NR′—Q (where R′ independently represents H or a lower alkyl group);
• X is C(R″)2,
• (wherein each R″ independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′ where n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R″), a tetrazole or oxadiazole of the form:
• wherein R′ is H or a lower alkyl group);
• or X is CR′═CR′, N═N, C═O, O, NR′, (where R′ represents H or a lower alkyl group), S, or SO2;
• each R1 and R2 independently is selected from the group consisting of H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SR′, COOR′, a tri-alkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined from R1 and R2), tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group), or a triazene of the form:
• (wherein R8 and R9 are lower alkyl groups) or
• and at least one of R2 is not H, OCH3, CH3 or halogen;
• each Q is independently selected from one of the following structures, each of which contains a carboxylic acid or an acid like functionality selected from the group consisting of hydroxy, sulfhydryl, tetrazole, oxadiazole, isoindol-1,3 (2H)-dione, benzisoxazole, 2,3-benzodiazine-1,4 (2H,3H)-dione, 2,3-benzoxazine-1,4 (3H)-dione, (2H) 1,3-benzoxazine-2,4 (3H)-dione, (3H) 2-benzazine-1,3 (2H)-dione, and NO2:
• IA, IB, IC, ID, IE, IF, and IG, wherein
• IA has the following structure:
• wherein:
• each of R3, R4, R5, R6, or R7 is independently defined the same as R1 above and wherein at least one of R3, R4, R5, R6, or R7 is a hydroxy, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• IB has the following structure:
• wherein:
• each of R10, R11, R12, R13, R14, R15, or R16 independently is defined the same as R1 above above and, wherein at least one of R10, R11, R12, R13, R14, R15, or R16 is a hydroxy, tetrazole, oxadiazole, NO2, or carboxy in both Q's and wherein at least one of R1 or R2 is a halogen when the compound of Formula I is a 4,4′-diazophenyl compound;
• IC has the following structure:
• wherein:
• each of R17, R18, R19, R20, or R21 independently is defined the same as R1 above;
• ID has the following structure:
• wherein each of R22, R23, or R24 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IE has the following structure:
• wherein:
• each of R25, R26, or R27 independently is defined the same as R1 above and
• represents a heterocyclic ring of one of the six following formulas:
• IF has the following structure:
• wherein:
• exactly one of R28, R29, R30, R31, or R32 is the
• of the
• link defined for Formula I above and each other R28, R29, R30, R31, or R32 independently is defined the same as R1 above, and wherein at least one of R28, R29, R30, R31, or R32 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• IG has the following structure:
• wherein:
• exactly one of R33, R34, R35, R36, R37, R38 or R39 is the
• of the
• link defined for Formula I above and each other R33, R34, R35, R36, R37, R38 or R39 independently is defined the same as R1 above and wherein at least one of R33, R34, R35, R36, R37, R38 or R39 is a hydroxy, sulfhydryl, tetrazole, oxadiazole, NO2, or carboxy in both Q's;
• or wherein:
• is NR′—N═Q (wherein R′ represents H or a lower alkyl group) and each
• is independently selected from one of the following structures:
• IIA or IIB, wherein:
• IIA has the following structure
• wherein
• each of R40-R47 independently is H, F, Cl, Br, I, a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, O—CH2—CH2F, CH2—CH2—CH2F, O—CH2—CH2—CH2F, CN, (C═O)—R′, N(R′)2, NO2, (C═O)N(R′)2, O(CO)R′, OR′, SH, COOR′, a tri-aLkyl tin, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tri-alkyl tin, a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group);
• IIB has the following structure:
• wherein:
• each of R48 and R49 independently is a lower alkyl group, (CH2)nOR′, wherein n=1, 2, or 3, CF3, CH2—CH2F, CH2—CH2—CH2F, CN, (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, (C═O)—CH2)n—(C6H5) where n=1, 2, or 3, Rph, CR′═CR′—Rph, CR2′—CR2′—Rph, (where Rph represents an unsubstituted or substituted phenyl group with the phenyl substituents being chosen from any of the non-phenyl substituents defined for R1), a tetrazole or oxadiazole of the form:
• (wherein R′ is H or a lower alkyl group,
• wherein at least one of R48 or R49 is (C═O)—R′, NO2, (C═O)N(R′)2, COOR′, CN, or a tetrazole or oxadiazole of the form:
• (wherein R′ is H or lower alkyl group) in both Q's.