Claims (20)
1. Konjugat hormona rasta s formulom (I):
A-W-B-GH (I)
naznačen time što
GH predstavlja hormon rasta kao spoj koji ima jednu mutaciju u Cys koju se bira iz skupine mutacija koje odgovaraju: T3C, P5C, S7C, D11C, H18C, Q29C, E30C, E33C, A34C, Y35C, K38C, E39C, Y42C, S43C, D47C, P48C, S55C, S57C, P59C, S62, E65C, Q69C, E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C, G126C, E129C, D130C, G131C, P133C, T135C, G136C, T142C, D147C, N149C, D154C, A155C, L156C, R178C, E186C, G187C i G190C u hGH (SEQ ID NO: 1);
B predstavlja hidrofilnu razmaknicu;
W je kemijska skupina koja spaja A s B, i
A predstavlja ostatak koji se veže na albumin, kojeg se bira između
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gdje * označuje vezanje na B preko W
gdje je ostatak koji se veže na albumin (A) preko hidrofilne razmaknice (B) spojen na GH preko sumpornog ostatka u jednoj mutaciji u Cys, ili njegova farmaceutski prihvatljiva sol.1. Growth hormone conjugate with formula (I):
A-W-B-GH (I)
indicated by what
GH represents growth hormone as a compound having one Cys mutation selected from the group of mutations corresponding to: T3C, P5C, S7C, D11C, H18C, Q29C, E30C, E33C, A34C, Y35C, K38C, E39C, Y42C, S43C, D47C . , T135C, G136C, T142C, D147C, N149C, D154C, A155C, L156C, R178C, E186C, G187C and G190C in hGH (SEQ ID NO: 1);
B represents a hydrophilic spacer;
W is the chemical group that joins A with B, i
A represents a residue that binds to albumin, which can be chosen between
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where * indicates binding to B via W
wherein the albumin-binding residue (A) via a hydrophilic spacer (B) is linked to GH via a sulfur residue in a single mutation to Cys, or a pharmaceutically acceptable salt thereof.
2. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što se jednu mutaciju u Cys bira između bilo koje od: T3C, P5C, S7C, D11C, H18C, Q29C, E30C, E33C, A34C, Y35C, E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C i G126C u hGH (SEQ ID NO: 1).2. Conjugate according to claim 1, characterized in that one mutation in Cys is selected from any of: T3C, P5C, S7C, D11C, H18C, Q29C, E30C, E33C, A34C, Y35C, E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C and G126C in hGH (SEQ ID NO: 1).
3. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što se jednu mutaciju u Cys bira između bilo koje od: E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C i G126C u hGH (SEQ ID NO: 1).3. Conjugate according to claim 1, characterized in that one mutation in Cys is selected from any of: E88C, Q91C, S95C, A98C, N99C, S100C, L101C, V102C, Y103C, D107C, S108C, D112C, Q122C and G126C in hGH (SEQ ID NO: 1).
4. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što je jedna mutacija u Cys L101C.4. Conjugate according to claim 1, characterized in that one mutation is in Cys L101C.
5. Konjugat u skladu s bilo kojim od prethodnih patentnih zahtjeva 1-4, naznačen time što GH ima dodatni disulfidni most.5. Conjugate according to any of the preceding claims 1-4, characterized in that GH has an additional disulfide bridge.
6. Konjugat u skladu s patentnim zahtjevom 5, naznačen time što GH ima dodatni disulfidni most između najmanje jednog od aminokiselinskih parova na položajima koji odgovaraju R16C/L117C, A17C/E174C, H21C/M170C, D26C/V102C, D26C/Y103C, N47C/T50C, Q49C/G161C, F54C/Y143C, F54C/S144C, F54C/F146C, S55C/Y143C, S57C/Y143C, 158C/Q141C, 158C/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 i/ili V185C/S188C u hGH (SEQ ID NO: 1).6. Conjugate according to claim 5, characterized in that GH has an additional disulfide bridge between at least one of the amino acid pairs at positions corresponding to R16C/L117C, A17C/E174C, H21C/M170C, D26C/V102C, D26C/Y103C, N47C/ T50C, Q49C/G161C, F54C/Y143C, F54C/S144C, F54C/F146C, S55C/Y143C, S57C/Y143C, 158C/Q141C, 158C/Y143C, I58C/S144C, P59C/Q137C, P61C/E66C, P61C/E66C, S71C/S132C, L73C/S132C, L73C/F139C, R77C/I138C, R77C/F139C, L81C/Q141C, L81C/Y143C, Q84C/Y143C, Q84C/S144C, S85C/Y143C, S85C/S144C, P46C/F92C, F92C/F1 F146C, F92C/T148C, R94C/D107C, V102C/A105C, L156C/F146C, L156C/T148C and/or V185C/S188C in hGH (SEQ ID NO: 1).
7. Konjugat u skladu s bilo kojim od prethodnih patentnih zahtjeva 5 i 6, naznačen time što je najmanje jedan od cisteina u dodatnoj disulfidnoj vezi prisutan u segmentu petlju, primjerice iz aminokiselinskih ostataka 128-154 (L3).7. Conjugate according to any of the previous patent claims 5 and 6, characterized in that at least one of the cysteines in the additional disulfide bond is present in the loop segment, for example from amino acid residues 128-154 (L3).
8. Konjugat u skladu s patentnim zahtjevom 7, naznačen time što dodatna disulfidna veza spaja L3 s uzvojnicom 2.8. Conjugate according to claim 7, characterized in that an additional disulfide bond joins L3 to helix 2.
9. Konjugat u skladu s bilo kojim od prethodnih patentnih zahtjeva 1-8, naznačen time što je GH najmanje 95% istovjetan s hGH (SEQ ID 1).9. Conjugate according to any of the preceding claims 1-8, characterized in that the GH is at least 95% identical to hGH (SEQ ID 1).
10. Konjugat u skladu s bilo kojim od patentnih zahtjeva 2-9, naznačen time što W ima formulu
-W7-Y-,
gdje
Y je -(CH2)l7-C3-10-cikloalkil-W8- ili valentna veza,
l7 je 0-6,
W7 se bira između -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH=CHC(O)-, -(CH2)s3-, -C(O)-, -C(O)O-, -OC(O)- ili valentne veze; gdje je s3 0 ili 1,
W8 se bira između -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH=CHC(O)-, -(CH2)s4-, -C(O)-, -C(O)O-, -OC(O)- ili valentne veze; gdje je s4 0 ili 1.10. A conjugate according to any one of claims 2-9, characterized in that W has the formula
-W7-Y-,
where
Y is -(CH2)17-C3-10-cycloalkyl-W8- or a valence bond,
l7 is 0-6,
W7 is selected from -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O )2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH =CHC(O)-, -(CH2)s3-, -C(O)-, -C(O)O-, -OC(O)- or valence bonds; where s3 is 0 or 1,
W8 is selected from -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O )2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH =CHC(O)-, -(CH2)s4-, -C(O)-, -C(O)O-, -OC(O)- or valence bonds; where s4 is 0 or 1.
11. Konjugat u skladu s bilo kojim od patentnih zahtjeva 2-10, naznačen time što B ima formulu
-X1-X2-X3-X4-,
gdje
X1 je -W1-[(CHR1)l1-W2]m1-{[(CH2)n1E1]m2-[(CHR2)l2-W3]m3}n2-,
X2 je -[(CHR3)l3-W4]m4-{[(CH2)n3E2]m5-[(CHR4)l4-W5]m6}n4-,
X3 je -[(CHR5)l5-W6]m7-,
X4 je F-D1-(CH2)l6-D2-,
l1, l2, l3, l4, l5 i l6 se neovisno bira između 0-16, primjerice od 0-6, m1, m3, m4, m6 i m7 se neovisno bira između 0-10, primjerice od 0-6, m2 i m5 se neovisno bira između 0-25, primjerice od 0-10, n1, n2, n3 i n4 se neovisno bira između 0-16, primjerice od 0-10, F je aril, hetaril, pirolidin-2,5-dion ili valentna veza, gdje arilna i hetarilna skupina mogu biti supstituirane s halogenom, -CN, -OH, -C(O)OH, -C(O)NH2, -S(O)2OH ili C1-6-alkilom,
R1, R2, R3, R4 i R5 se neovisno bira između vodika, -C(O)OH, -C(O)NH2, -S(O)OH, -S(O)2OH, -NHC(=NH)-NH2, C1-6-alkila, arila ili hetarila; gdje alkilna, arilna i hetarilna skupina mogu biti supstituirane s halogenom, -C(O)OH, -C(O)NH2, -S(O)OH, -S(O)2OH, -CN ili -OH,
D1, D2, E1 i E2 se neovisno bira između -O-, -N(R6)-, -N(C(O)R7)- ili valentne veze; gdje R6 i R7 neovisno predstavljaju vodik ili C1-6-alkil, W1 do W5 se neovisno bira između -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH=CHC(O)-, -(CH2)s2-, -C(O)-, -C(O)O-, -OC(O)- ili valentne veze; gdje je s2 0 ili 1,
W6 se bira između -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH=CHC(O)-, -(CH2)s1-, -C(O)-, -C(O)O-, -OC(O)-, -NHC(O)C1-6-alkil, -C(O)NHC1-6-alkila ili valentne veze; gdje je s1 0 ili 1, a C1-6-alkilna skupina može biti supstituirana s okso, pirolidin-2,5-dionom, -NHC(O)CH*CH2COOH ili -NHC(O)CH2CH*COOH;
gdje (*) ukazuje na mjesto vezanja od atoma ugljika u CH na X4.11. A conjugate according to any one of claims 2-10, characterized in that B has the formula
-X1-X2-X3-X4-,
where
X1 is -W1-[(CHR1)l1-W2]m1-{[(CH2)n1E1]m2-[(CHR2)l2-W3]m3}n2-,
X2 is -[(CHR3)13-W4]m4-{[(CH2)n3E2]m5-[(CHR4)14-W5]m6}n4-,
X3 is -[(CHR5)l5-W6]m7-,
X4 is F-D1-(CH2)16-D2-,
l1, l2, l3, l4, l5 and l6 are independently chosen between 0-16, for example from 0-6, m1, m3, m4, m6 and m7 are independently chosen between 0-10, for example from 0-6, m2 and m5 is independently selected from 0-25, for example from 0-10, n1, n2, n3 and n4 is independently selected from 0-16, for example from 0-10, F is aryl, hetaryl, pyrrolidine-2,5-dione or valence bond, where aryl and hetaryl groups can be substituted with halogen, -CN, -OH, -C(O)OH, -C(O)NH2, -S(O)2OH or C1-6-alkyl,
R1, R2, R3, R4 and R5 are independently selected from hydrogen, -C(O)OH, -C(O)NH2, -S(O)OH, -S(O)2OH, -NHC(=NH)- NH 2 , C 1-6 -alkyl, aryl or hetaryl; where alkyl, aryl and hetaryl groups can be substituted with halogen, -C(O)OH, -C(O)NH2, -S(O)OH, -S(O)2OH, -CN or -OH,
D1, D2, E1 and E2 are independently selected from -O-, -N(R6)-, -N(C(O)R7)- or a valence bond; where R6 and R7 independently represent hydrogen or C1-6-alkyl, W1 to W5 are independently selected from -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)- , -C(O)NHS(O)2-, -S(O)2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C (O)-, -C(O)CH=CH-, -CH=CHC(O)-, -(CH2)s2-, -C(O)-, -C(O)O-, -OC(O )- or valence bonds; where s2 is 0 or 1,
W6 is selected from -C(O)NH-, -NHC(O)-, -C(O)NHCH2-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O )2NHC(O)-, -OC(O)NH-, -NHC(O)O-, -C(O)CH2-, -CH2C(O)-, -C(O)CH=CH-, -CH =CHC(O)-, -(CH2)s1-, -C(O)-, -C(O)O-, -OC(O)-, -NHC(O)C1-6-alkyl, -C( O)NHC1-6-alkyl or valence bonds; where s1 is 0 or 1, and the C1-6-alkyl group may be substituted with oxo, pyrrolidine-2,5-dione, -NHC(O)CH*CH2COOH or -NHC(O)CH2CH*COOH;
where (*) indicates the binding site from the carbon atom in CH to X4.
12. Konjugat u skladu s patentnim zahtjevom 11, naznačen time što se D1 i D2 neovisno bira između -O- ili - N(R6)- ili valentne veze.12. Conjugate according to claim 11, characterized in that D1 and D2 are independently selected from -O- or -N(R6)- or a valence bond.
13. Konjugat u skladu s patentnim zahtjevom 11 ili 12, naznačen time što se E1 i E2 neovisno bira između -O- ili -N(R6)- ili valentne veze.13. Conjugate according to claim 11 or 12, characterized in that E1 and E2 are independently selected from -O- or -N(R6)- or a valence bond.
14. Konjugat u skladu s bilo kojim od patentnih zahtjeva 10-13, naznačen time što se W1 do W8 neovisno bira iz skupine koju čine -C(O)NH-, -NHC(O)-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -NHC(O)C1-6-alkil ili -C(O)NHC1-6-alkil ili valentna veza; gdje alkilna skupina može biti supstituirana s okso, pirolidin-2,5-dionom, -NHC(O)CH*CH2COOH ili -NHC(O)CH2CH*COOH; gdje (*) ukazuje na mjesto vezanja od atoma ugljika u CH na X4.14. Conjugate according to any of claims 10-13, characterized in that W1 to W8 are independently selected from the group consisting of -C(O)NH-, -NHC(O)-, -CH2NHC(O)-, -C(O)NHS(O)2-, -S(O)2NHC(O)-, -NHC(O)C1-6-alkyl or -C(O)NHC1-6-alkyl or a valence bond; where the alkyl group may be substituted with oxo, pyrrolidine-2,5-dione, -NHC(O)CH*CH2COOH or -NHC(O)CH2CH*COOH; where (*) indicates the binding site from the carbon atom in CH to X4.
15. Konjugat u skladu s bilo kojim od patentnih zahtjeva 11-14, naznačen time što se R1, R2, R3, R4 i R5 neovisno bira između vodika, -C(O)OH, -C(O)NH2, -S(O)2OH ili C1-6-alkila; gdje alkilna skupina može biti supstituirana s -C(O)OH, -C(O)NH2 ili -S(O)2OH.15. Conjugate according to any of claims 11-14, characterized in that R1, R2, R3, R4 and R5 are independently selected from hydrogen, -C(O)OH, -C(O)NH2, -S( O) 2 OH or C 1-6 -alkyl; where the alkyl group may be substituted with -C(O)OH, -C(O)NH2 or -S(O)2OH.
16. Konjugat u skladu s bilo kojim od patentnih zahtjeva 11-15, naznačen time što je X4 valentna veza, a W6 se bira između bilo pirolidin-2,5-diona, -NHC(O)CH*CH2COOH ili -NHC(O)CH2CH*COOH, gdje (*) ukazuje na mjesto vezanja od atoma ugljika u CH na GH.16. A conjugate according to any one of claims 11-15, characterized in that X4 is a valence bond and W6 is selected from either pyrrolidine-2,5-dione, -NHC(O)CH*CH2COOH or -NHC(O )CH2CH*COOH, where (*) indicates the attachment site from the carbon atom in CH to GH.
17. Konjugat u skladu s bilo kojim od patentnih zahtjeva 2-16, naznačen time što se B bira između
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.17. A conjugate according to any one of claims 2-16, characterized in that B is selected from
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18. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što se navedeni konjugat bira između
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.18. Conjugate according to claim 1, characterized in that said conjugate is chosen between
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19. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što se navedeni konjugat bira između
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.19. Conjugate according to claim 1, characterized in that said conjugate is chosen between
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20. Konjugat u skladu s patentnim zahtjevom 1, naznačen time što se navedeni konjugat bira između
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.20. Conjugate according to claim 1, characterized in that said conjugate is chosen between
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