Conjugated proteins with prolonged in vivo efficacy
The invention relates to conjugated proteins, in particular but not exclusively, blood coagulation factors, to processes for preparing the conjugated proteins which contain the steps of reacting a protein or glycoprotein, such as factor VIIa or human growth hormone, with a water insoluble albumin binder in the presence of an optionally substituted cyclodextrin molecule, to pharmaceutical compositions comprising the protein conjugates and to the use of the protein conjugates in therapy, in particular but not exclusively, for the treatment of diseases alleviated by blood coagulation factors such as the prophylactic treatment of hemophilia.
1. A process for preparing a conjugated blood coagulation factor which comprises the steps of reacting the blood coagulation factor with a water insoluble albumin binder in the presence of an optionally substituted cyclodextrin molecule, wherein said water insoluble albumin binder comprises a —(CH 2 ) 12 — moiety.
2. The process of claim 1 , wherein the cyclodextrin molecule comprises β-cyclodextrin, optionally substituted by one or more C 1-6 alkyl, each of which may be optionally substituted by one or more hydroxyl groups.
3. The process of claim 1 , comprising reacting in an aqueous buffered solution.
4. The process of claim 1 , wherein said blood coagulation factor is selected from the group consisting of FVII, FX, FII, FV, protein C, protein S, tPA, PAI-1, tissue factor, FXI, FXII, FXIII, FVIII sequence variants, and FIX sequence variants.
5. A blood coagulation factor conjugate selected from the group consisting of
6. The process of claim 2 , wherein the optionally substituted cyclodextrin is 2-hydroxyethyl-β-cyclodextrin.
7. The process of claim 3 , wherein the aqueous buffered solution is a Hepes buffer.
8. The process of claim 7 , wherein the Hepes buffer comprises 50 mM Hepes, 100 mM NaCl and 10 mM CaCl 2 .
9. The process of claim 4 , wherein said coagulation factor is FVII.