Labelled derivatives of steroids

Claim 2

A compound having the structural formula ##STR8## wherein one or two groups Z are =NO(CHR) n CO.sup.. R 1 , any remaining group Z being = 0; X is --OH or --H; r is H or C 1 to C 4 alkyl; n is 1, 2 or 3; R 1 is either --OH or --NH.sup.. CHY.sup.. (CH 2 ) m Q; y is --H or --COOR 2 ; m is 0, 1, 2, 3, 4 or 5; R 2 is H, CH 3 or C 2 H 5 ; q is unlabelled or labelled and has one of the following structural formulae: ##STR9##

Claim 3

A compound as claimed in claim 1, wherein R is H; n is 1; and R 1 is --OH.

Claim 4

A compound as claimed in claim 1, wherein R is H; n is 1; R 1 is --NH.sup.. CHY.sup.. (CH 2 ) m Q; m is 1; and Q is ##STR10##

Claim 5

A compound as claimed in claim 1, labelled with one or more 125-iodine atoms.

Claim 6

A compound as claimed in claim 4, wherein X is H; r is H; n is 1; R 1 is --NH.sup.. CH(CO.sup.. OCH 3 )CH 2 Q q is ##STR11##

Claim 7

A compound as claimed in claim 4, wherein X is OH; r is H; n is 1; R 1 is --NH.sup.. CH(COOCH 3 ) CH 2 Q; q is ##STR12##

Claim 8

A process for making a compound as claimed in claim 1, wherein R 1 is --OH, which process consists essentially of a. oxidising digoxin or digitoxin to the dialdehyde; b. reacting the dialdehyde with an o-carboxyalkoxyamine having the formula NH.sub.2.sup. . O(CHR).sub.n COOH where r and n are as defined in claim 1, to form a mono- or di-o-carboxyalkyloxime derivative thereof.

Claim 9

A process for making a compound as claimed in claim 1 wherein R 1 is --NH.sup.. CHY.sup.. (CH 2 ) m Q, which process consists essentially of the steps: a. oxidising digoxin or digitoxin to the dialdehyde; b. reacting the dialdehyde with an o-carboxyalkoxylamine having the formula NH.sub.2.sup.. O(CHR).sub.n COOH where R and n are as defined in claim 1, to form a mono- or di-o-carboxyalkyloxime derivative thereof, c. reacting the resulting oxime derivative by formation of a peptide bond, in the presence of an N,N'-substituted carbodimide, with an amine having the formula NH 2 .sup.. CHY.sup.. (CH 2 ) m Q, where Y, m and Q are as defined in claim 1, to form an amine conjugate thereof.

Claim 10

A process as claimed in claim 8, comprising the additional step of radio-iodinating the resulting amine conjugate.

Claim 11

In a method of performing a competitive radio-assay for digoxin or digitoxin by the steps of a. causing the digoxin or digitoxin to compete with a standard amount of radioactively labelled digoxin or digitoxin for reaction with a specific reagent therefor present in a standard amount insufficient to bind all the labelled and unlabelled compound, b. separating the bond compound from the unbound compound; and c. measuring the proportion of radioactivity in at least one of the bound and the unbound compound, the improvement wherein the radioactively labelled digoxin or digitoxin is an amine conjugate of a digoxin or digitoxin dialdehyde bis-o-carboxyalkyl oxime having the structural formula ##STR13## wherein one or two groups Z are =NO(CHR) n CO.sup.. R 1 , any remaining group Z being = 0; X is --OH or --H; r is H or C 1 to C 4 alkyl; n is 1, 2 or 3; R 1 is either --OH or --NH.sup.. CHY.sup.. (CH 2 ) m Q; y is --H or --COOR 2 ; m is 0, 1, 2, 3, 4 or 5; R 2 is H, CH 3 or C 2 H 5 ; q is unlabelled or labelled and has one of the following structural formulae: ##STR14##

Claim 12

In a method of performing a competitive radio-assay for digoxin or digitoxin by the steps of a. causing the digoxin or digitoxin to compete with a standard amount of radioactively labelled digoxin or digitoxin for reaction with a specific reagent therefor present in a standard amount insufficient to bind all the labelled and unlabelled compound, b. separating the bond compound from the unbound compound; and c. measuring the proportion of radioactivity in at least one of the bound and the unbound compound, the improvement wherein the radioactively labelled digoxin or digitoxin is an amine conjugate of a digoxin or digitoxin dialdehyde bis-o-carboxyalkyl oxime having the structural formula ##STR13## wherein one or two groups Z are =NO(CHR) n CO.sup.. R 1 , any remaining group Z being = 0; X is --OH or --H; r is H or C 1 to C 4 alkyl; n is 1, 2 or 3; R 1 is either --OH or --NH.sup.. CHY.sup.. (CH 2 ) m Q; y is --H or --COOR 2 ; m is 0, 1, 2, 3, 4 or 5; R 2 is H, CH 3 or C 2 H 5 ; q is unlabelled or labelled and has one of the following structural formulae: ##STR14##

Claim 13

A method as claimed in claim 10, wherein the radioactive labelled digoxin or digitoxin is a compound is labelled with one or more I 125 atoms.