DK2621519T3 - Leptin-ABD fusionspolypeptider med forbedret virkningsvarighed - Google Patents

Leptin-ABD fusionspolypeptider med forbedret virkningsvarighed Download PDF

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DK2621519T3
DK2621519T3 DK11833080.2T DK11833080T DK2621519T3 DK 2621519 T3 DK2621519 T3 DK 2621519T3 DK 11833080 T DK11833080 T DK 11833080T DK 2621519 T3 DK2621519 T3 DK 2621519T3
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amino acid
leptin
acid sequence
modified polypeptide
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Soumitra S Ghosh
Mary Erickson
David C Litzinger
Zijian Guo
Jonathan David Roth
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Aegerion Pharmaceuticals Inc
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Claims (33)

1. Modificeret polypeptid omfattende: et albuminbindende domæne polypeptid (ABD), som omfatter et albuminbindende motiv (ABM), omfattende aminosyresekvensen: GVSD X5 YK X8 X91 X11 X12 A Xi4 TVEGV X20 AL X23 X24 X251 (SEQ ID NO: 34), hvor, uafhængigt af hinanden, X5 er valgt blandt Y og F; Xe er valgt blandt N, R og S; X9 er valgt blandt V, I, L, M, F og Y; X11 er valgt blandt N, S, E og D; Xi2 er valgt blandt R, K og N; X14 er valgt blandt K og R; X20 er valgt blandt D, N, Q, E, H, S, R og K; X23 er valgt blandt K, I og T; X24 er valgt blandt A, S, T, G, H, L og D; og X25 er valgt blandt Η, E og D; og et første peptidhormondomæne (HD1) med en aminosyresekvens valgt blandt: (a) SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, aminosyrer 2-147 ifølge SEQ ID NO:31, og aminosyrer 2-147 ifølge SEQ ID NO:33; eller (b) aminosyresekvenserne 1-146 for et leptin valgt blandt: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22 SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145 og SEQ ID NO:146; hvori en afvigende aminosyre er substitueret i én eller flere af følgende positioner idet nummereringen fastholdes (selv under fraværet af en glutaminylrest ved position 28): 4, 32, 33, 35,50,64,68,71,74,77,78, 89,97, 100, 102, 105, 106, 107, 108, 111, 118, 136, 138, 142 og 145; (c) aminosyresekvensen ifølge underafsnit (b), i hvilken glutaminylresten ved position 28 er fraværende; (d) aminosyresekvensen ifølge underafsnit (b) eller (c), i hvilken en methionylrest er tilføjet ved N-terminus; (e) en leptin bestående af et fragment af aminosyresekvensen ifølge (b), (c) eller (d) valgt blandt: i) aminosyrerne 98-146; ii) aminosyrerne 1-32; iii) aminosyrerne 40-116; iv) aminosyrerne 1-99 og 112-146; v) aminosyrerne 1-99 og 112-146, hvor én eller flere af aminosyrerne 100- 111 er placeret imellem aminosyrerne 99 og 112; vi) aminosyresekvensen ifølge underafsnit (i), hvor én eller flere af aminosyrerne 100, 102, 105, 106, 107, 108, 111,118, 136, 138, 142 og 145 er substitueret med en anden aminosyre; vii) aminosyresekvensen ifølge underafsnit (ii), hvor én eller flere af aminosyrerne 4, 8 og 32 er substitueret med en anden aminosyre; viii) aminosyresekvensen ifølge underafsnit (iii), hvor én eller flere af aminosyrerne 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111 og 112 er erstattet med en anden aminosyre; ix) aminosyresekvensen ifølge underafsnit (iv), hvor én eller flere af aminosyrerne 4, 8, 32, 33, 35, 48, 50, 53, 60, 64, 66, 67, 68, 71, 74, 77, 78, 89, 97, 112, 118, 136, 138, 142 og 145 er erstattet med en anden aminosyre; og x) aminosyresekvensen ifølge underafsnit (v), hvor én eller flere af aminosyrerne 4, 32, 33, 35, 50, 64, 68, 71, 74, 77, 78, 89, 97, 100, 102, 105, 106, 107, 108, 111, 118, 136, 138, 142 og 145 er erstattet med en anden aminosyre; xi) aminosyresekvensen ifølge ethvert af underafsnittene (i)-(x), hvor et methionin er tilføjet ved N-terminus; (f) aminosyresekvensen ifølge ethvert af undersnittene (b) til og med (e), hvor aminosyresekvensen er forbundet med en kemisk gruppe; (g) aminosyresekvensen ifølge underafsnit (f), hvor den kemiske gruppe er en vandopløselig polymergruppe; (h) aminosyresekvensen ifølge underafsnit (g), hvor den vandopløselige polymergruppe er valgt blandt polyethylenglycol, en ethylenglycol/propylenglycol-copolymer, en carboxymethylcellulose, en dextran, en polyvinylalkohol, en polyvinylpyrrolidon, en poly-1,3-dioxolan, en poly-1,3,6-trioxan, en ethylen/maleinanhydrid-copolymer, en polyaminosyre-homopolymer, en tilfældig sammensat polyaminosyre-copolymer, en albumin, et Fc-protein, en poly(n-vinyl pyrrolidon)polyethylenglycol, en propylenglycol-homopolymer, en polypropylenoxid/ethylenoxid-copolymer, en polyoxyethyleret polyol, en polyvinylalkohol, et polyethylenglycolpropionaldehyd, et succinat og en styren; (i) aminosyresekvens ifølge underafsnit (h), hvor den vandopløselige polymergruppe er en polyethylenglycol; og (j) aminosyresekvens ifølge underafsnit (h), hvor den vandopløselige polymer er en polyaminosyre valgt blandt: en albumin, et antistof, et Fc-protein og en polylysingruppe; og en første linker (L1) kovalent forbundet med den nævnte HD1, idet linkeren omfatter en aminosyresekvens valgt blandt Gly-Gly-Gly, [Gly-Ser]n, [Gly-Gly-Ser]„, [Gly-Gly-Gly-Ser]n og [Gly-Gly-Gly-Gly-Ser]n, hvor n er 1, 2, 3, 4, 5, 6, 7, 8, 9 eller 10.
2. Modificeret polypeptid ifølge krav 1, hvor HD1 har en sekvens valgt blandt sekvenserne ifølge underafsnit (a).
3. Modificeret polypeptid ifølge krav 2, hvor det modificerede polypeptid omfatter det nævnte ABD som en N-terminal gruppe, og det nævnte HD1 som en C-terminal gruppe.
4. Modificeret polypeptid ifølge krav 2, hvor det modificerede polypeptid omfatter det nævnte ABD som en C-terminal gruppe og det nævnte HD1 som en N-terminal gruppe.
5. Modificeret polypeptid ifølge ethvert af kravene 1 til 4, hvor det nævnte HD1 har en aminosyresekvens valgt blandt: SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID N0:31, SEQ ID NO:32, SEQ ID NO:33, aminosyrerne 2-147 ifølge SEQ ID NO:31, og aminosyrerne 2-147 ifølge SEQ ID NO:33.
6. Modificeret polypeptid ifølge ethvert af kravene 1 til 5, hvor det nævnte HD1 har en aminosyresekvens valgt blandt: SEQ ID NO:26, SEQ ID NO:29, SEQ ID NO:33 og aminosyrerne 2-147 ifølge SEQ ID NO:33.
7. Modificeret polypeptid ifølge ethvert af kravene 1 til 6, hvor, uafhængigt af hinanden, X5erY; X8 er N; X23 erT eller I; X24 er S eller L; og X25 er E eller H.
8. Modificeret polypeptid ifølge ethvert af kravene 1 til 7, hvor det albuminbindende motiv omfatter en aminosyresekvens, som er valgt blandt: GVSDYYKNLINKAKTVEGVEALTLHI (SEQ ID NO:114) og GVSDYYKNLINKAKTVEGVEALISEI (SEQ ID NO:115).
9. Modificeret polypeptid ifølge ethvert af kravene 1 til 8, hvor det nævnte ABD omfatter et albuminbindende motiv (ABM), som ikke er GVSDYYKNLINNAKTVEGVKALIDEI (SEQ ID NO:35).
10. Modificeret polypeptid ifølge ethvert af kravene 1 til 9, hvor det albuminbindende domæne polypeptid (ABD) omfatter en aminosyresekvens, som er valgt blandt: LAEAKEDAIKELDKYGVSDYYKNLINKAKTVEGVEALTLHILAALP (SEQ ID NO:50); og LAEAKEDAIKELDKYGVSDYYKNLINKAKTVEGVEALISEILAALP (SEQ ID NO:51).
11. Modificeret polypeptid ifølge krav 1 eller 2, hvor det nævnte ABD omfatter am inosyresekvensen: LAEAK Xa Xb A Xc Xd EL Xe KY -[ABM]- LAALP (SEQ ID NO:36) hvor [ABM] er det albuminbindende motiv, og, uafhængigt af hinanden, Xa er valgt blandt V og E; Xb er valgt blandt L, E og D; Xc er valgt blandt N, L og I; Xd er valgt blandt R og K; Xe er valgt blandt D og K; leucinet ved position 45 foreligger eller mangler; og prolinet ved position 46 foreligger eller mangler.
12. Modificeret polypeptid ifølge krav 1, hvor det nævnte ABD omfatter en amino-syresekvens med i det mindste 85% identitet med en aminosyresekvens, som er valgt blandt: SEQ ID NO: 37, SEQ ID NO:39, SEQ ID NO:40, SEQ ID NO:41, SEQ ID NO:43, SEQ ID NO:44, SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:48, SEQ ID NO:49, SEQ ID NO:50, SEQ ID NO:51 og SEQ ID NO:52.
13. Modificeret polypeptid ifølge krav 1 eller 2, hvor det nævnte ABD omfatter et vilkårligt af peptiderne valgt blandt: LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVNALTHHILAALP (SEQ ID NO:39), LAEAKVLANRELDKYGVSDYYKNLINRARTVEGVHALIDHILAALP (SEQ ID NO:40), LAEAKVLANRELDKYGVSDYYKNIINRAKTVEGVRALKLHILAALP (SEQ ID NO:41), LAEAKVLANRELDKYGVSDYYKNLINKAKTVEGVEALTLHILAALP (SEQ ID NO:43), LAEAKVLANRELDKYGVSDFYKNLINRAKTVEGVDALIAHILAALP (SEQ ID NO:44), LAEAKVLANRELDKYGVSDFYKSLINRAKTVEGVDALTSHILAALP (SEQ ID NO:45), LAEAKVLANRELDKYGVSDFYKNVINKAKTVEGVEALIADILAALP (SEQ ID NO:47), LAEAKVLANRELDKYGVSDYYKNLINKAKTVEGVQALIAHILAALP (SEQ ID NO:48), LAEAKVLANRELDKYGVSDFYKRLINKAKTVEGVEALKLHILAALP (SEQ ID NO:49), LAEAKEDAIKELDKYGVSDYYKNLINKAKTVEGVEALTLHILAALP (SEQ ID NO:50), LAEAKEDAIKELDKYGVSDYYKNLINKAKTVEGVEALISEILAALP (SEQ ID NO:51) og LAEAKEDAIKELDKYGVSDYYKRLISKAKTVEGVKALISEILAALP (SEQ ID NO:52).
14. Modificeret polypeptid ifølge krav 1 eller 2, hvor det nævnte ABD omfatter aminosyresekvensen ifølge SEQ ID NO:49.
15. Modificeret polypeptid ifølge ethvert af kravene 1 til 14, hvor linkeren L1 er et peptid med fra 1 til 30 aminosyrer eller mindre end 30 aminosyrer.
16. Modificeret polypeptid ifølge ethvert af kravene 1 til 15, hvor linkeren L1 omfatter et N-terminal TG dipeptid eller et C-terminal AS dipeptid, eller begge.
17. Modificeret polypeptid ifølge ethvert af kravene 1 til 16, hvor linkeren L1 omfatter en aminosyresekvens, som er valgt blandt: TG-(GGGS)i (SEQ ID NO: 215), TG-(GGGS)2 (SEQ ID NO: 216), TG (GGGS)3 (SEQ ID NO: 217), TG-(GGGS)4 (SEQ ID NO: 218), TG-(GGGS)5 (SEQ ID NO: 219), (GGGS)rAS (SEQ ID NO: 220), (GGGS)2-AS (SEQ ID NO: 221), (GGGS)3-AS (SEQ ID NO: 222), (GGGS)4-AS (SEQ ID NO: 223), (GGGS)s-AS (SEQ ID NO: 224), TG-(GGGS)i-AS (SEQ ID NO: 225), TG-(GGGS)2-AS (SEQ ID NO: 226), TG-(GGGS)3-AS (SEQ ID NO: 227), TG-(GGGS)4-AS (SEQ ID NO: 228) og TG-(GGGS)5-AS (SEQ ID NO: 229).
18. Modificeret polypeptid ifølge ethvert af kravene 1 til 17, hvor det modificerede polypeptid omfatter en aminosyresekvens valgt blandt: SEQ ID NO:53, SEQ ID NO:54, SEQ ID NO:55, SEQ ID NO:56, SEQ ID NO:57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72, SEQ ID NO:73, SEQ ID NO:74, SEQ ID NO:75, SEQ ID NO:76, SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:85 SEQ ID NO:86, SEQ ID NO:87, SEQ ID NO:88, SEQ ID NO:89, SEQ ID NO:90, SEQ ID NO:91, SEQ ID NO:92, SEQ ID NO:93, SEQ ID NO:94, SEQ ID NO:95, SEQ ID NO:96, SEQ ID NO:97, SEQ ID NO:98, SEQ ID NO:99, SEQ ID NO:100, SEQ ID NO:101, SEQ ID NO:102, SEQ ID NO:103, SEQ ID NO:104, SEQ ID NO:105, SEQ ID NO:106 og SEQ ID NO:107.
19. Modificeret polypeptid ifølge krav 18, hvor det modificerede polypeptid omfatter en aminosyresekvens valgt blandt: SEQ ID NO:54 og SEQ ID NO:61.
20. Modificeret polypeptid ifølge krav 1, hvor det nævnte HD1 har en sekvens valgt blandt sekvenserne i underafsnit (b)-(j).
21. Modificeret polypeptid ifølge krav 20, hvor det nævnte HD1 omfatter en aminosyresekvens valgt blandt: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NQ:30, SEQ ID N0:31, SEQ ID NO:32, SEQ ID NO:33, SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145 og SEQ ID NO:146; hvori én eller flere aminosyresubstitutioner er foretaget.
22. Modificeret polypeptid ifølge krav 19, hvor det nævnte HD1 er SEQ ID NO:143, SEQ ID NO:144, SEQ ID NO:145 eller SEQ ID NO:146.
23. Modificeret polypeptid ifølge krav 20, hvor det nævnte HD1 er SEQ ID NO:26, SEQ ID NO:29 eller SEQ ID NO:33, hvori én eller flere aminosyresubstitutioner er foretaget.
24. Modificeret polypeptid ifølge ethvert af kravene 20 til 23, hvor det nævnte ABD omfatter aminosyresekvensen ifølge SEQ ID NO:49.
25. Modificeret polypeptid ifølge ethvert af kravene 1 til 24 til anvendelse ved behandling af en sygdom eller lidelse i et subjekt.
26. Modificeret polypeptid ifølge ethvert af kravene 1 til 24 til anvendelse ved behandling af en sygdom eller lidelse valgt blandt lipodystrofi, dyslipidemi, hyperlipidemi, obesitet, hypothalmisk amenorrhe, Alzheimer's sygdom, leptindeficiens, fedtleversygdom, diabetes (inkl. type I og type II), ikke-alkoholisk steatohepatitis (NASH), ikke-alkoholisk fedtleversygdom (NAFLD), metabolisk syndrom X og Huntington's sygdom.
27. Modificeret polypeptid til anvendelse ifølge krav 26, hvor sygdommen eller lidelsen er lipodystrofi, dyslipidemi, hyperlipidemi, obesitet, hypothalmisk amenorrhe, Alzheimer's sygdom, leptindeficiens, fedtleversygdom eller diabetes.
28. Farmaceutisk sammensætning omfattende et modificeret polypeptid ifølge ethvert af kravene 1 til 24 og et farmaceutisk acceptabelt excipiens.
29. Farmaceutisk sammensætning ifølge krav 28, hvor den farmaceutiske sammensætning er en injicerbar farmaceutisk sammensætning.
30. Farmaceutisk sammensætning ifølge krav 28 eller krav 29, hvor den farmaceutiske sammensætning er en farmaceutisk sammensætning med forsinket frigivelse eller langvarig virkning.
31. Farmaceutisk sammensætning ifølge ethvert af kravene 28 til 30 til anvendelse ved behandling af lipodystrofi, dyslipidemi, hyperlipidemi, obesitet, hypothalmisk amenorrhe, Alzheimer's sygdom, leptindeficiens, fedtleversygom, diabetes (inkl. type I og type II), ikke-alkoholisk steatohepatitis (NASH), ikke-alkoholisk fedtleversygdom (NAFLD) eller metabolisk syndrom X.
32. Kombination af i det mindste to forskellige mod obesitet virkende midler til anvendelse ved behandling af obesitet i et subjekt, hvor i det mindste ét imod obesitet virkende middel er en amylin-konjugeret til polyethylenglycol (PEG) og i det mindste ét mod obesitet virkende middel er et modificeret polypeptid ifølge ethvert af kravene 1 til 24.
33. Den ikke-terapeutiske anvendelse af en kombination af i det mindste to forskellige mod obesitet virkende midler til reduktion af kropsvægt i et subjekt, hvor i det mindste ét imod obesitet virkende middel er en amylin-konjugeret til polyethylenglycol (PEG) og i det mindste ét mod obesitet virkende middel er et modificeret polypeptid ifølge ethvert af kravene 1 til 24.
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