Near infrared radiation in-vivo diagnostic methods and dyes

• US 6,083,485 A
• Filed: 11/07/1997
• Issued: 07/04/2000
• Est. Priority Date: 12/07/1994
• Status: Expired due to Term

• Current Assignee: VisEn Medical, Inc. (Revvity, Inc.)
• Original Assignee: Der Freien Universitat Berline, Institut fur Diagnostikforschung GmbH
• Inventors: Licha, Kai, Speck, Ulrich, Semmler, Wolfhard, Hilger, Christoph-Stephan, Riefke, Bjorn
• Primary Examiner(s): Dees, Jose' G.
• Assistant Examiner(s): JONES, DAMERON LEVEST
• Application Number:
  US08/849,369
• Time in Patent Office: 970 Days
• Field of Search: 424/1.11, 424/1.65, 424/9.1, 424/9.3, 424/9.4, 424/9.5, 424/9.6, 424/1.69, 540/1, 544/1, 544/3, 544/63, 548/100, 548/146, 548/215, 548/400, 548/300.1, 568/300, 568/303, 568/700, 546/152, 8/DIG. 13
• US Class Current: 424/9.6
• CPC Class Codes: A61K 49/0017 Fluorescence in vivo
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0032 Methine dyes, e.g. cyanine dyes
• CPC Class Codes: A61K 49/005 characterised by the carrier molecule carrying the fluorescent agent
• CPC Class Codes: C09B 23/086 more than five >CH- groups
• CPC Class Codes: C09B 69/105 containing a methine or polymethine dye

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.

Claims

1. An in-vivo diagnostic near infrared radiation method comprising:

(a) administering a compound of the general formula I

space="preserve" listing-type="equation">B.sub.1 --(F--W.sub.n).sub.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 having the general formula IIb ##STR10## wherein R³ to R¹² are same or different, each independently representing a hydrogen atom, a residue B or W, or an alkyl or alkenyl residue containing up to 6 carbon atoms or an aryl or aralkyl residue containing up to 9 carbon atoms, said alkyl, alkenyl, aryl or aralkyl residue optionally carrying an additional residue W or W to each pair of adjacent residues R³ to R¹⁰ are annealed, with due regard for the interspersed atoms, 5- to 6-member rings that may be saturated, unsaturated or aromatic, and that may optionally carry residue R,

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 residue containing up to 6 carbon atoms, an aryl, alkylaryl hydroxyaryl, carboxyaryl, sulfoaryl, arylamino, diarylamino, nitroaryl or halogenaryl residue containing up to 9 carbon atoms,

s and t independently represent the numbers 0 to 1 on the condition that s and t do not represent the number 1 at the same time,

X and Y are same or different, each independently representing an O, S, Se or Te or a --C(CH₃)₂ --, --CH═

CH-- or --CR¹³ R¹⁴ -- fragment,

wherein R¹³ and R¹⁴ independently represent a hydrogen atom, a residue B or W, or an alkyl or alkenyl residue containing up to 6 carbon atoms or an aryl or aralkyl residue containing up to 9 carbon atoms, the alkyl, alkenyl, aryl or aralkyl residue optionally having an additional residue W;

W represents a hydrophilic group that improves water-solubility, with the 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 their physiologically tolerable salts;

(b) irradiating the compound with light in the near infrared range; and

(c) recording the radiation produced by the compound.

2. A method according to claim 1, wherein B within the general formula I is selected from the group consisting of an amino acid, a peptide, a 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.

3. A method according to claim 1, wherein W within the general formula I is 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 residue of the general formula III

space="preserve" listing-type="equation">--(CH.sub.2).sub.a --O--Z or (--CH.sub.2 --CH.sub.2 --O).sub.a --Z(III)

wherein

a represents the numbers 0 to 6;

Z represents a hydrogen atom or an alkyl residue containing 3 to 6 C atoms that includes 2 to n-1 hydroxy groups, with n being the number of C atoms, or an aryl or aralkyl residue containing 6 to 10 C atoms and having 2 to 4 additional hydroxy groups, or an alkyl residue containing 1 to 6 C atoms and having 1 to 3 additional carboxy groups, or an aryl residue containing 6 to 9 C atoms and having 1 to 3 additional carboxyl groups, or an aralkyl residue or a nitroaryl or a nitroaralkyl residue containing 6 to 15 C atoms, or a sulfoalkyl residue containing 2 to 4 C atoms having 1 to 3 additional carboxy groups,

or represents a residue of the general formula IIIa or IIIb ##STR11## or a residue of the general formula IIIc

space="preserve" listing-type="equation">--(CH.sub.2).sub.o --(CO).sub.p --NR.sup.1 --(CH.sub.2).sub.s --(NH--CO).sub.q --R.sup.2 (IIIc)

wherein

o and s independently represent the numbers, 0, 1, 2, 3, 4, 5 or 6,

p and q independently represent 0 to 1,

R¹ and R² independently represent a residue Z except the substituents of the general formulas IIIa and IIIb, or independently represent a residue of the general formulas IIId or IIIe;

##STR12## on the condition that p and q=1; and

(v) a residue of the general formula IIIc.

4. A cyanine dye of the general formula V ##STR13## where Q is a fragment selected from the group consisting of ##STR14## and ##STR15## where R³⁰ represents a hydrogen atom, a hydroxy group, a carboxy group, an alkoxy residue containing 1 to 4 carbon atoms or a chlorine atom, b is an integer 2 or 3, R³¹ represents a hydrogen atom or an alkyl residue containing 1 to 4 carbon atoms,

X and Y independently represent an --O--, --S--, --CH═

CH-- or --C(CH₂ R³²)(CH₂ R³³)-- fragment each,

R²⁰ to R²⁹, R³² and R³³ independently represent a hydrogen atom, a hydroxy group, a carboxy-, a sulfonic acid residue or a carboxyalkyl-, alkoxycarbonyl or alkoxyoxoalkyl residue containing up to 10 C atoms

or a sulfoalkyl residue containing up to 4 C atoms, or a non-selectively bonding macromolecule or a residue of the general formula VI

space="preserve" listing-type="equation">--(O).sub.v --(CH.sub.2).sub.o --CO--NR.sup.34 --(CH.sub.2).sub.s --(NH--CO).sub.q --R.sup.35 (VI)

on the condition that, where X and Y are O, S, --CH═

CH-- or --C(CH₃)₂ -- at least one of the residues R²⁰ to R²⁹ corresponds to a non-selectively bonding macromolecule or a compound of the general formula VI, where

o and s equal 0 or independently represent an integer between 1 and 6,

q and v independently represent 0 or 1,

R³⁴ represents a hydrogen atom or a methyl residue,

R³⁵ represents an alkyl residue containing 3 to 6 C atoms and comprising 2 to n-1 hydroxy groups, with n being the number of C atoms, or an alkyl residue containing 1 to 6 C atoms that carries 1 to 3 additional carboxy groups, an aryl residue containing 6 to 9 C atoms or arylalkyl residue containing 7 to 15 C atoms, or a residue of the general formula IIId or IIIe ##STR16## on the condition that q is 1, or a non-selectively bonding macromolecule,

R²⁰ and R²¹, R²¹ and R²², R²² and R²³, R²⁴ and R²⁵, R²⁵ and R²⁶, R²⁶ and R²⁷, together with the interspersed carbon atoms, form a 5- or 6-member aromatic or saturated annelled ring,

and its physiologically tolerable salts.

5. An agent for in-vivo diagnosis comprising the cyanine dye according to claim 4 together with physiologically acceptable adjuvants, substrates, and/or diluents.

6. A cyanine dye according to claim 4, wherein said dye is selected from the group consisting of

5-[2-[(1,2-dicarboxyethyl)amino]-2-oxoethyl]-2-[7-[5-[2-[(1,2-dicarboxyethyl)amino]-2-oxoethyl]-1,3-dihydro-3,3-dimethyl-1-(4-sulfobutyl)-2H-indol-2-yliden]-1,3,5-heptatrienyl]-3,3-dimethyl-1-(4-sulfobutyl)-3H-indolium, inner salt, potassium hydrogen salt;

2-[7-[5-[2-[(11-carboxy-2-oxo-1,4,7,10-tetraaza-4,7,10-tri(carboxymethyl)-1-undecyl)amino]-2-oxoethyl]-1,3-dihydro-3,3-dimethyl-1-ethyl-2H-indol-2-yliden]-1,3,5-heptatrienyl]-3,3-dimethyl-1-(4-sulfobutyl)-3H-indolium, inner salt;

2-[7-[1,3-dihydro-3,3-dimethyl-5-[2-[(methoxypolyoxyethylene)-amino]-2-oxoethyl]-1-(4-sulfobutyl)-2H-indol-2-yliden]-1,3,5-heptatrienyl]-3,3-dimethyl-5-[2-[(methoxypolyoxyethylene)amino]-2-oxoethyl]-1-(4-sulfobutyl)-3H-indolium, sodium salt;

2-[7-[1,3-dihydro-3,3-dimethyl-1-(4-sulfobutyl)-2H-indol-2-yliden]-1,3,5-heptatrienyl]-3,3-dimethyl-5-(methoxypolyoxy-ethylene)aminocarbonyl-1-(4-sulfobutyl)-3H-indolium, sodium salt;

3-(3-carboxypropyl)-2-[7-[3-(3-carboxypropyl)-1,3-dihydro-3-methyl-1-(4-sulfobutyl)-2H-indol-2-yliden]-1,3,5-heptatrienyl]-3-methyl-1-(4-sulfobutyl)-3H-indolium, sodium salt;

2-[[3-[[3-(3-carboxypropyl)-1,3-dihydro-3-methyl-1-(4-sulfobutyl)2H-indol-2-yliden]methyl]-2-hydroxy-4-oxo-2-cyclobuten-1-yliden]methyl]-1,1-dimethyl-3-ethyl-1H-2-benz(e)indolium, inner salt and;

2-[7-[1,3-dihydro-5-[2-[(2,3-dihydroxypropyl)amino]-2-oxoethyl]-3,3-dimethyl-1-(4-sulfobutyl)-2H-indol-2-yliden]-1,3,5-heptatrienyl]-5-[2-[(2,3-dihydroxypropyl)amino]-2-oxoethyl]-3,3-dimethyl-1-(4-sulfobutyl)-3H-indolium, sodium salt.

7. A method of diagnosing diseased tissue comprising administering the cyanine dye claimed in claim 4, irradiating the diseased tissue with light in the visible to near infrared range and recording fluorescent radiation produced from the cyanine dye.