Polypeptide and protein derivatives and process for their preparation
View Patent ↗Protein and polypeptid derivatives and their salts are claimed characterized in that a protein or polypeptide is conjugated via an intermediate grouping containing at least one radical of the formula —C(R)═N— (or —N═C(R)—) or —CH(R)—NH— (or —NH—CH(R)—), wherein R is hydrogen or a hydrocarbon residue which may be substituted, with the same or a different protein or polypeptide, with a reporter group or a cytotoxic agent as well as a process for their preparation and the novel intermediates therefor.
1. A method of producing a composition containing modified protein or polypeptide molecules, or salts thereof, wherein said modified protein or polypeptide molecules of said composition consist essentially of a compound selected from
A-X-Z-X′—B
wherein
A is a residue of a protein or polypeptide having a carboxy and amino terminus and is connected to X-Z-X′—B exclusively at said carboxy or amino terminus;
B is a polymeric compound;
X and X′ independently from each other are bivalent organic radicals or independently from each other are present or absent;
Z is a bivalent radical selected from the group consisting of: —C(R)═N—, —N═C(R)—, —CH(R)—NH—, —NH—CH(R)—, —C(R)═N—Y—NC(R)—, —N═C(R)—Y—C(R)N—, —CH(R)—NH—Y—NH—CH(R)— and —NH—CH(R)—Y—CH(R)—NH—, —C(R)═N—O—, —O—N═C(R)—, —CH(R)—NH—O—, —O—NH—CH(R)—, —C(R)═N—O—Y—O—N═C(R)—, —O—N═C(R)—Y—C(R)═N—O—, —CH(R)—NH—O—Y—O—NH—CH(R)— and —O—NH—CH(R)—Y—CH(R)—NH—O—;
where
R is hydrogen or an aliphatic, cycloaliphatic, aromatic or araliphatic hydrocarbon group; and
Y is a bivalent organic group,
wherein said method comprises condensing a compound of the formula:
A-X—R 1
wherein R 1 is a —CO—R group, an acetalized formyl group, or an amino or protected amino group, and A,R, and X are as defined above, with a compound of formula:
R 2 —X′—B
or a compound of formula:
R 2 —Y—R 2
where R 2 is amino when R 1 is —CO—R or acetalized formyl and R2 is —CO—R or acetalized formyl when R 1 is amino, and X′, Y, R and B are as defined above, to form a Schiff base, hydrazone, oxime or azomethine compound, and optionally,
reducing the —C(R)═N— or —N═C(R) formed by the condensation to CH(R)—NH— or NH—CH(R)—, respectively, and optionally forming a salt.
2. The method of producing a composition of claim 1 , wherein said residue A is a carboxy terminal residue.
3. The method of producing a composition of claim 1 , wherein said residue A is an amino terminal residue.
4. The method of producing a composition of claim 1 , wherein R is hydrogen.
5. The method of producing a composition of claim 1 , wherein said polymeric compound B comprises a compound selected from the group consisting of:
(i) desferioxamine B, or a metal derivative thereof
(ii) diethylenetriaminepentaacetic acid, or a metal derivative thereof
(iii) [Nε-(diethylenetriaminepentaacetic acid -alanyl)-Lys]5, or a metal derivative thereof; and
(iv) a polyglutamic acid having at least two ferioxamine B residues coupled thereto.
6. The method of producing a composition of claim 1 , wherein Z is —CH 2 —NH—, or —NH—CH 2 —.
7. The method of producing a composition of claim 1 , wherein Z is —C(R)═N—, or —N═C(R)—.
8. The method of producing a composition of claim 1 , wherein Z is —CH(R)—NH—, or —NH—CH(R)—.
9. The method of producing a composition of claim 1 , wherein Z is —C(R)═N—O— or —O—N═C(R)—.
10. The method of producing a composition of claim 1 , wherein Z is CH(R)—NH—O—, —O—NH—CH(R)—.
11. The composition of claim 1 , wherein Z is —C(R)═N—Y—N═C(R)—, —N═C(R)—Y—C(R)═N—, —CH(R)—NH—Y—NH—CH(R)— or —NH—CH(R)—Y—CH(R)—NH—.
12. The composition of claim 1 , wherein Z is —CH═N—Y—N═CH—, —N═CH—Y—CH═N—, —CH 2 —N—Y—N—CH 2 —, or —NH—CH 2 —Y—CH 2 —NH—.
13. The composition of claim 1 , wherein Z is —C(R)═N—O—Y—O—N═C(R)—, —O—N═C(R)—Y—C(R)═N—O—.
14. The composition of claim 1 , wherein Z is —CH(R)—NH—O—Y—O—NH—CH(R)— or —O—NH—CH(R)—Y—CH(R)—NH—O—.
15. The method of producing a composition of claim 1 , wherein said polymeric compound B is a protein or polypeptide that is the same or different from said protein or polypeptide A, or is a reporter group or cytotoxic agent.
16. The method of producing a composition of claim 15 , wherein said polymeric compound B is a protein or polypeptide that is the same as said protein or polypeptide A.
17. The method of producing a composition of claim 15 , wherein said polymeric compound B is a protein or polypeptide that is different from said protein or polypeptide A.
18. The method of producing a composition of claim 15 , wherein said polymeric compound B is a cytotoxic agent.
19. The method of producing a composition of claim 15 , wherein said polymeric compound B is a reporter group.
20. The method of producing a composition of claim 19 , wherein said polymeric compound B is a reporter group comprising a metal chelating organic compound.