US 12,264,191 B2
De-immunized factor VIII molecule and pharmaceutical compositions comprising the same
Karina Winterling, Darmstadt (DE); Steffen Kistner, Frankfurt am Main (DE); Jens Daufenbach, Mainz (DE); Annie de Groot, Providence, RI (US); William Martin, Providence, RI (US); and Christopher Ungerer, Langen (DE)
Assigned to Biotest AG, Dreieich (DE)
Appl. No. 17/046,601
Filed by Biotest AG, Dreieich (DE)
PCT Filed Apr. 11, 2019, PCT No. PCT/EP2019/059233
§ 371(c)(1), (2) Date Oct. 9, 2020,
PCT Pub. No. WO2019/197524, PCT Pub. Date Oct. 17, 2019.
Claims priority of application No. 18166982 (EP), filed on Apr. 12, 2018.
Prior Publication US 2024/0010707 A1, Jan. 11, 2024
Int. Cl. C07K 14/755 (2006.01); A61K 31/573 (2006.01); A61K 31/675 (2006.01); A61K 38/37 (2006.01); A61K 39/395 (2006.01); C07K 16/28 (2006.01); G01N 33/50 (2006.01)
CPC C07K 14/755 (2013.01) [A61K 31/573 (2013.01); A61K 31/675 (2013.01); A61K 38/37 (2013.01); A61K 39/39541 (2013.01); C07K 16/2887 (2013.01); G01N 33/5047 (2013.01); G01N 33/505 (2013.01)] 29 Claims
 
1. A recombinant Factor VIII protein comprising at least three amino acid substitutions at positions selected from the group consisting of Y748, L171, S507, N79, I80, I105, S112, L160, V184, N233, L235, V257, I265, N299, Y426, Y430, L505, F555, I610, N616, I632, L706, N754, K1837, R1936, S2030, S2037, N2038, S2077, M2123, S2125, F2215, K2226, K2258, V2313, S2315, V2333 and Q2335;
wherein substitutions of N are independently selected from the group consisting of D, H, S and E; wherein substitution of I are independently selected from the group consisting of T and V; wherein substitutions of S are independently selected from the group consisting of A, N, G, T and E; wherein substitutions of L are independently selected from the group consisting of N, Q, F and S; wherein substitutions of V are independently selected from the group consisting of A and T; wherein substitutions of Y are independently selected from the group consisting of N, H and S; wherein substitutions of F are independently selected from the group consisting of H and S; wherein substitutions of K are independently selected from the group consisting of N, D, E, Q, S and T; wherein substitutions of R are independently selected from the group consisting of Q, H and S; wherein substitutions of M are selected from the group consisting of R, Q, K and T; and/or wherein substitutions of Q are selected from the group consisting of R, D, E, H and K;
wherein the positions are specified in relation to full length human Factor VIII molecule of SEQ ID NO: 1;
and wherein the recombinant Factor VIII protein retains at least 50% coagulant activity, as determined in a chromogenic assay, compared to a Factor VIII protein consisting of SEQ ID NO: 2,
or a fusion protein of said recombinant Factor VIII protein.