In-vivo diagnostic method by near infrared radiation

Abstract

This invention relates to an in-vivo diagnostic method based on near infrared radiation (NIR radiation) that uses water-soluble dyes and their biomolecule adducts, each having specific photophysical and pharmaco-chemical properties, as a contrast medium for fluorescence and transillumination diagnostics in the NIR range, to new dyes and pharmaceuticals containing such dyes.

1 - 8 . (canceled)  
     
     
         9 . An in-vivo diagnostic near infrared radiation method comprising: 
 (a) administering a compound of the general formula I      B l -(F-W n ) m   (I)     wherein 
 l represents a number from 0 to 6, n represents a number from 0 to 10 and m represents a number from 1 to 100;  
 B is a biological detecting unit having a molecular weight of up to 30,000 that bonds to specific cell populations or selectively to receptors, or accumulates in tissues or tumours, or generally stays in the blood, or is a macromolecule that bonds non-selectively;  
 F represents a dye showing maximum absorption in the range of 650 to 1200 nm;  
 W represents a hydrophilic group that improves water-solubility, with n-octanol-water distribution coefficient of the compound according to formula I being less than or equal to 2.0 for l=0; as well as a physiologically tolerable salt; W being selected from the group consisting of (i) a carboxy or sulfonic acid group; (ii) a carboxyalkyl group; (iii) an alkoxycarbonyl or alkoxyoxoalkyl group containing up to 12 carbon atoms, (iv) a group of the general formula IIII  
   —(CH 2 ) a —O-Z or (—CH 2 —CH 2 —O) a -Z  (III)  
  wherein  
 a represents the numbers 0 to 6;  
 Z represents a hydrogen atom or an alkyl group containing 3 to 6 C atoms having 2 to n−1 hydroxy groups, with n being the number of C atoms or an aryl or aralkyl group containing 6 to 10 C atoms and having 2 to 4 additional hydroxy groups, or an alkyl group containing 1 to 6 C atoms and having 1 to 3 additional carboxy groups, or an aryl group containing 6 to 9 C atoms and having 1 to 3 additional carboxyl groups or an aralkyl group or a nitroaryl or a nitroaralkyl group containing 6 to 15 C atoms or a sulfoalkyl group containing 2 to 4 C atoms; a group of the general formulas III a or III b  
                     
 a group of the general formula IIIc  
   —(CH 2 ) o —(CO) p —NR 1 —(CH 2 ) s —(NH—CO) q —R 2   (IIIc)  
  wherein  
 o and s independently represent the numbers 0, 1, 2, 3, 4, 5 or 6,  
 p and q independently represent 0 or 1,  
   R 1  and NR 2  independently represent a group Z except the substituents of the general formulas IIIa and IIIb, or independently represent a group of the general formulas IIId or IIIe                         wherein p and q=1; and      (b) a group of the general formula III as defined above;    (c) irradiating the compound with light in the near infrared range; and    (d) recording the radiation produced by the compound, wherein F represents a cyanine dye of the general formula IIa                           wherein    r represents the numbers 0, 1 or 2, wherein, for r=2, the respective L 6  and L 7  groups that occur in duplicate may be the same or different, L 1  to L 7  are the same or different, each independently representing a CH or CR group,    where    R is a halogen atom, a hydroxy, carboxy acetoxy, amino, nitro, cyano or sulfonic acid group or an alkyl, alkenyl, hydroxyalkyl, carboxyalkyl, alkoxy, alkoxycarbonyl, sulfoalkyl, alkylamino, dialkylamino or halogenalkyl group containing up to 6 carbon atoms, aryl, alkylaryl, hydroxyaryl, carboxyaryl, sulfoaryl, arylamino, diarylamino, nitroaryl or halogenaryl group containing up to 9 carbon atoms, or where R represents a bond that bonds to another group R and forms a 4- to 6-member ring together with the interspersed groups L 1  to L 7 , or two of L 1  to L 7  that are linked via a —CO— group,    R 3  to R 12  are the same or different, each independently representing a hydrogen atom, a group B or W, or an alkyl or alkenyl group containing up to 6 carbon atoms or an aryl or aralkyl group optionally carying an additional group W, or to each pair of adjacent groups R 3  to R 10  are annealed, with due regard for the interspersed C atoms, 5- to 6-member rings that may be saturated, unsaturated or aromatic, and that optionally carry an additional group R, and    X and Y are the same or different, each independently representing an O, S, Se or Te or a —C(CH 3 ) 2 —, —CH═CH— or —CR 13 R 14 — group,    where R 13  and R 14  independently represent a hydrogen atom, a group B or W, or an alkyl or alkylene group containing up to 6 carbon atoms or an aryl or aralkyl group containing up to 9 carbon atoms, the alkyl, alkenyl, aryl or aralkyl group optionally carrying an additional group W.    
     
     
         10 . A method according to  claim 9 , wherein B within the general formula I is selected from the group consisting of an amino acid, a peptide, CDR (complementarity determining region), an antigen, a hapten, an enzyme substrate, an enzyme cofactor, biotin, a carotinoid, a hormone, a neurohormone, a neurotransmitter, a growth factor, a lymphokin, a lectin, a toxin, a carbohydrate, an oligosaccharide, a polysaccharide, a dextran, an oligonucleotide, and a receptor-bonding pharmaceutical.