Targeted modified TNF family members
The present invention relates to a modified cytokine of the TNF superfamily, with reduced activity to its receptor, wherein said modified cytokine is specifically delivered to target cells. Preferably, said modified cytokine is a single chain variant of the TNF superfamily, even more preferably, one or more of the chains can-y one or more mutations, resulting in a low affinity to the receptor, wherein said mutant cytokine is specifically delivered to target cells. The targeting is realized by fusion of the modified cytokine of the TNF superfamily to a targeting moiety, preferably an antibody or antibody-like molecule. The invention relates further to the use of such targeted modified cytokine of the TNF superfamily to treat diseases.
1. A composition comprising a proteinaceous construct, comprising
(i) a single chain polypeptide comprising three modified human TNFs, wherein:
each modified human TNF comprises a modified amino acid residue by substitution at the I97 position relative to wild type human TNF (SEQ ID NO: 14), the substitution being selected from A, Q and S
and the modified human TNFs have reduced affinity towards their receptor as compared to wild type human TNF; and
(ii) a targeting moiety that is an antibody or a variable domain of a camelid heavy chain antibody (VHH) directed to a neo-vasculature tissue or cancer tissue specific marker,
wherein the composition has significant biological activity towards cells that are targeted by the targeting moiety.
2. The composition of claim 1 , wherein the targeting moiety is a VHH.
3. The composition of claim 1 , wherein the targeting moiety is directed towards CD20.
4. The composition of claim 1 , wherein the targeting moiety is directed towards Her2.
5. The composition of claim 3 , wherein the targeting moiety is a VHH.
6. The composition of claim 4 , wherein the targeting moiety is a VHH.
7. The composition of claim 1 , wherein the 197 substitution is I97A.
8. The composition of claim 1 , wherein the 197 substitution is I97Q.
9. The composition of claim 1 , wherein the 197 substitution is I97S.
10. The composition of claim 5 , wherein the 197 substitution is I97A.
11. The composition of claim 5 , wherein the 197 substitution is I97Q.
12. The composition of claim 5 , wherein the 197 substitution is I97S.
13. The composition of claim 6 , wherein the 197 substitution is I97A.
14. The composition of claim 6 , wherein the 197 substitution is I97Q.
15. The composition of claim 6 , wherein the 197 substitution is I97S.