Methods and compositions for targeting proteins across the blood brain barrier
View Patent ↗Disclosed are methods and compositions for targeting therapeutic proteins to the brain. Methods and compositions of the invention involve associating an IGF moiety with a therapeutic protein in order to target the therapeutic protein to the brain. Soluble fusion proteins that include an IGF targeting moiety are transported to neural tissue in the brain from blood. Methods and compositions of the invention include therapeutic applications for treating lysosomal storage diseases. The invention also provides nucleic acids and cells for expressing IGF fusion proteins.
1. A targeted therapeutic fusion protein comprising: a lysosomal enzyme; and a mutein of human IGF-I having an amino acid sequence at least 80% identical to mature human IGF-I (SEQ ID NO:1); wherein the mutein of IGF-I comprises an amino acid substitution selected from the group consisting of leucine at a position corresponding to Tyr 60 , leucine at a position corresponding to Tyr 24 , serine at a position corresponding to Phe 16 , and lysine at a position corresponding to Glu 9 , or amino acid substitutions at positions corresponding to Arg 55 and Arg 56 , of mature human IGF-I (SEQ ID NO:1); and wherein the mutein binds to the human cation-independent mannose-6-phosphate receptor in a mannose-6-phosphate-independent manner.
2. The targeted therapeutic fusion protein of claim 1 , wherein the mutein of human IGF-I comprises an amino acid substitution of leucine at a position corresponding to Tyr 60 of mature human IGF-I (SEQ ID NO:1).
3. The targeted therapeutic fusion protein of claim 1 , wherein the mutein of human IGF-I comprises an amino acid substitution of leucine at a position corresponding to Tyr 24 of mature human IGF-I (SEQ ID NO:1).
4. The targeted therapeutic fusion protein of claim 1 , wherein the mutein of human IGF-I comprises an amino acid substitution of serine at a position corresponding to Phe 16 of mature human IGF-I (SEQ ID NO:1).
5. The targeted therapeutic fusion protein of claim 1 , wherein the mutein of human IGF-I comprises an amino acid substitution of lysine at a position corresponding to Glu 9 of mature human IGF-I (SEQ ID NO:1).
6. The targeted therapeutic fusion protein of claim 1 , wherein the mutein of human IGF-I comprises amino acid substitutions at positions corresponding to Arg 55 and Arg 56 of mature human IGF-I (SEQ ID NO:1).
7. A targeted therapeutic fusion protein comprising: a lysosomal enzyme; and a mutein of human IGF-I having an amino acid sequence at least 80% identical to mature human IGF-I (SEQ ID NO:1); wherein the mutein binds to the human cation-independent mannose-6-phoshate receptor in a mannose-6-phoshate-independent manner and wherein the fusion protein is deglycosylated.
8. The targeted therapeutic fusion protein of claim 6 , wherein the fusion protein is deglycosylated by periodate treatment.