Growth hormones with prolonged in-vivo efficacy

1. A growth hormone conjugate which comprises a growth hormone compound (GH) comprising a single Cys mutation selected from the group consisting of E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C, and G126C, numbered according to SEQ ID NO: 1,

wherein an albumin binding residue via a hydrophilic spacer is linked to the sulphur residue of said single Cys of said GH, and wherein said albumin binding residue comprises a linear lipophilic moiety containing 12-40 carbon atoms, or a pharmaceutically acceptable salt thereof.

2. The growth hormone conjugate of claim 1 wherein the growth hormone conjugate has the formula (I):

A-W—B-GH  (I)

wherein

GH represents the growth hormone compound;

B represents a hydrophilic spacer linked to the sulphur residue of said single Cys of said GH;

W is a chemical group linking A and B; and

A represent an albumin binding residue comprising a linear lipophilic moiety containing 12-40 carbon atoms;

and pharmaceutically acceptable salts thereof.

3. The growth hormone conjugate of claim 1 , wherein the GH further comprising an additional disulfide bridge between at least one of the amino acid pairs in the positions selected from the group consisting of R16C/L117C, A17C/E174C, H21C/M170C, D26C/V102C, D26C/Y103C, N47C/T50C, Q49C/G161C, F54C/Y143C, F54C/S144C, F54C/F146C, S55C/Y143C, S57C/Y143C, I58C/Q141C, I58C/Y143C, I58C/S144C, P59C/Q137C, P61C/E66C, P61C/T67C, S71C/S132C, L73C/S132C, L73C/F139C, R77C/I138C, R77C/F139C, L81C/Q141C, L81C/Y143C, Q84C/Y143C, Q84C/S144C, S85C/Y143C, S85C/S144C, P89C/F146C, F92C/F146C, F92C/T148C, R94C/D107C, V102C/A105C, L156C/F146C, L156C/T148C and V185C/S188C in SEQ ID NO: 1.

4. The growth hormone conjugate of claim 2 , wherein A is selected from the group consisting of:

wherein * denotes the attachment to B through W.

5. The growth hormone conjugate of claim 2 , wherein W has the formula

—W 7 —Y—,

wherein

Y is —(CH 2 ) 17 —C 3-10 -cycloalkyl-W 8 — or a valence bond;

I7 is 0-6;

W 7 is selected from —C(O)NH—, —NHC(O)—, —C(O)NHCH 2 —, —CH 2 NHC(O)—, —C(O)NHS(O) 2 —, —S(O) 2 NHC(O)—, —OC(O)NH—, —NHC(O)O—, —C(O)CH 2 —, —CH 2 C(O)—, —C(O)CH═CH—, —CH═CHC(O)—, —(CH 2 ) s3 —, —C(O)—, —C(O)O—, —OC(O)—, or a valence bond;

wherein s3 is 0 or 1;

W 8 is selected from —C(O)NH—, —NHC(O)—, —C(O)NHCH 2 —, —CH 2 NHC(O)—, —C(O)NHS(O) 2 —, —S(O) 2 NHC(O)—, —OC(O)NH—, —NHC(O)O—, —C(O)CH 2 —, —CH 2 C(O)—, —C(O)CH═CH—, —CH═CHC(O)—, —(CH 2 ) s4 —, —C(O)—, —C(O)O—, —OC(O)—, or a valence bond;

wherein s4 is 0 or 1.

6. The growth hormone conjugate of claim 2 , wherein B has the formula

—X 1 —X 2 —X 3 —X 4 —

wherein

X 1 is —W 1 —[(CHR 1 ) I1 —W 2 ] m1 —{[(CH 2 ) n1 E1] m2 —[(CHR 2 ) I2 —W 3 ] m3 } n2 —;

X 2 is —[(CHR 3 ) I3 —W 4 ] m4 —{[(CH 2 ) n3 E2] m5 —[(CHR 4 ) I4 —W 5 ] m6 } n4 —;

X 3 is —[(CHR 5 ) I5 —W 6 ] m7 —;

X 4 is F-D1-(CH 2 ) I6 -D2-;

I1, I2, I3, I4, I5 and I6 independently are selected from 0-16, such as from 0-6

m1, m3, m4, m6 and m7 independently are selected from 0-10, such as from 0-6

m2 and m5 independently are selected from 0-25, such as from 0-10

n1, n2, n3 and n4 independently are selected from 0-16, such as from 0-10

F is aryl, hetaryl, pyrrolidine-2,5-dione or a valence bond, wherein the aryl and

hetaryl groups are optionally substituted with halogen, —CN, —OH, —C(O)OH, —C(O)NH 2 , —S(O) 2 OH or C 1-6 -alkyl;

R 1 , R 2 , R 3 , R 4 and R 5 independently are selected from hydrogen, —C(O)OH, —C(O)NH 2 , —S(O)OH, —S(O) 2 OH, —NH—C(═NH)—NH 2 , C 1-6 -alkyl, aryl or hetaryl, wherein the alkyl, aryl and hetaryl groups optionally are substituted with halogen, —C(O)OH, —C(O)NH 2 , —S(O)OH, —S(O) 2 OH, —CN or —OH;

D1, D2, E1 and E2 independently are selected from —O—, —N(R 6 )—, —N(C(O)R 7 )— or a valence bond, wherein R 6 and R 7 independently represent hydrogen or C 1-6 -alkyl;

W 1 to W 5 independently are selected from —C(O)NH—, —NHC(O)—, —C(O)NHCH 2 —, —CH 2 NHC(O)—, —C(O)NHS(O) 2 —, —S(O) 2 NHC(O)—, —OC(O)NH—, —NHC(O)O—, —C(O)CH 2 —, —CH 2 C(O)—, —C(O)CH═CH—, —CH═CHC(O)—, —(CH 2 ) s2 —, —C(O)—, —C(O)O—, —OC(O)—, or a valence bond, wherein s2 is 0 or 1;

W 6 is selected from —C(O)NH—, —NHC(O)—, —C(O)NHCH 2 —, —CH 2 NHC(O)—, —C(O)NHS(O) 2 —, —S(O) 2 NHC(O)—, —OC(O)NH—, —NHC(O)O—, —C(O)CH 2 —, —CH 2 C(O)—, —C(O)CH═CH—, —CH═CHC(O)—, —(CH 2 ) s1 —, —C(O)—, —C(O)O—, —OC(O)—, —NHC(O)C 1-6 -alkyl, —C(O)NHC 1-6 -alkyl or a valence bond, wherein s1 is 0 or 1 and the C 1-6 -alkyl group is optionally substituted with oxo, pyrrolidine-2,5-dione, —NHC(O)CH*CH 2 COOH or —NHC(O)CH 2 CH*COOH, wherein (*) indicates the attachment point from the carbon atom of CH to X 4 .

7. The growth hormone conjugate of claim 6 , wherein D1 and D2 are independently selected from —O— or —N(R 6 )— or a valence bond.

8. The growth hormone conjugate of claim 6 , wherein E1 and E2 are independently selected from —O— or —N(R 6 )— or a valence bond.

9. The growth hormone conjugate of claim 5 , wherein W 7 and W 8 independently are selected from the group consisting of —C(O)NH—, —NHC(O)—, —CH 2 NHC(O)—, —C(O)NHS(O) 2 —, —S(O) 2 NHC(O)—, —NHC(O)C 1-6 -alkyl or —C(O)NHC 1-6 -alkyl or a valence bond, wherein the alkyl group is optionally substituted with oxo, pyrrolidine-2,5-dione, —NHC(O)CH*CH 2 COOH or —NHC(O)CH 2 CH*COOH, wherein (*) indicates the attachment point from the carbon atom of CH to B.

10. The growth hormone conjugate of claim 6 , wherein R 1 , R 2 , R 3 , R 4 and R 5 independently are selected from hydrogen, —C(O)OH, —C(O)NH 2 , —S(O) 2 OH or C 1-6 -alkyl; wherein the alkyl group optionally is substituted with —C(O)OH, —C(O)NH 2 or —S(O) 2 OH.

11. The growth hormone conjugate of claim 6 , wherein X 4 is a valence bond and W 6 is selected from either pyrrolidine-2,5-dione, —NHC(O)CH*CH 2 COOH or —NHC(O)CH 2 CH*COOH, wherein (*) indicates the attachment point from the carbon atom of CH to GH.

12. The growth hormone conjugate of claim 2 wherein B is selected from the group consisting of:

13. A growth hormone conjugate selected from the group consisting of:

14. The growth hormone conjugate of claim 1 wherein said single Cys mutation is L101C numbered according to SEQ ID NO: 1.