Methods of manufacture and synthesis of fluorescent dye compounds and uses thereof
The present disclosure relates to synthesizing and utilizing fluorescent dye compounds or pharmaceutically acceptable salts thereof. Conjugation of amino acid groups to the fluorescent dyes increase specificity and detection of the compound. Methods of manufacture and synthesis of the compounds for use thereof in diagnostic imaging are contemplated.
1 . A method for synthesizing a compound of the formula selected from the group consisting of:
wherein:
R 1 is (CH 2 ) n and n is 1, 2, or 3;
R 2 is a halogen- or X-R 3 , wherein X is (CH 2 ) m , NH, O, S, or Se;
m is 0, 1, 2, or 3;
R 3 is [CH 2 ] p CH(R 4 )COOH or C 6 H 4 —[CH 2 ] p CH(R 4 )COOH or (CH 2 ) p COOH or (CH 2 ) p NH 2 and p is 0, 1, 2, or 3;
R 4 is H or NHR 5 ;
R 5 is H or alkyl group;
R 6 , R 7 are independently selected from the group consisting of CH 2 , O, and a combination thereof;
R 8 is (CH 2 ) q SO 3 H and q is 0, 1, 2, or 3; and
A is a counter cation selected from the group consisting of sodium, potassium, calcium, magnesium, lithium, cholinate, lysinium, ammonium, and hydrogen;
comprising the steps of:
(a) reacting a compound of a formula:
with a compound of a formula:
in the presence of an acetate salt to generate a compound of the formula:
(b) reacting the resulting compound with a compound of the formula:
in the presence of a non-polar solvent to generate a compound of the formula:
(c) reacting a compound of the formula:
in the presence of DMF, POCl 3 , and aniline to generate a compound of the formula:
(d) reacting the compound of the formula:
with a
compound of the formula:
in the presence of an acetate salt to generate a compound of Formula II:
2 . The method of claim 1 , further comprising step (e) wherein the compound of the Formula II:
is reacted with X-R 3 in polar solvent, wherein X=OH, NH 2 , SH, or SeH to generate a compound of the formula:
3 . The method of claim 2 , further comprising reacting the resulting compound of step (e) with a coupling agent and a compound of the formula Ligand-Linker-Z, wherein the Ligand is selected from:
(i) an antibody or antibody fragment selected from Fab, F(ab′)2, and Fv;
(ii) an amino acid or an amino acid derivative; or
(iii) a pteridin-containing small molecule; and
wherein Z is COOH, NH 2 , OH, or SH.
4 . The method of claim 1 , wherein the resulting compound is precipitated using a non-polar solvent or purified by chromatographic methods or crystallization methods.
5 . The method of claim 2 , wherein the resulting compound is precipitated using a non-polar solvent or purified by chromatographic methods or crystallization methods.
6 . The method of claim 1 , wherein the non-polar solvent or polar solvent is an organic solvent.
7 . The method of claim 2 , wherein the non-polar solvent or polar solvent is an organic solvent.
8 . The method of claim 1 , wherein the yield of the resulting compound in step (d) is at least 60%.
9 . The method of claim 2 , wherein the yield of the resulting compound in step (e) is at least 60%.
10 . The method of claim 1 , wherein the purity of the resulting compound in step (d) is at least 90%.
11 . The method of claim 2 , wherein the purity of the resulting compound in step (e) is at least 90%.
12 . The method of claim 1 or claim 2 , wherein the resulting compound is selected from the group consisting of:
13 . The method of claim 12 , wherein the resulting compound is: