Mannose-6-phosphate bearing peptides fused to lysosomal enzymes
A targeted therapeutic including a lysosomal enzyme and a lysosomal targeting moiety that is a peptide containing at least one N-linked glycosylation site. Methods of producing the targeted therapeutic may include nucleotide acids encoding the same and host cells co-expressing GNPT. Pharmaceutical compositions comprising the targeted therapeutic and methods of using the same to treat a lysosomal storage disease.
1. A nucleic acid encoding a targeted therapeutic fusion protein comprising:
a lysosomal enzyme, and
a lysosomal targeting moiety,
wherein the lysosomal targeting moiety is a prosaposin peptide comprising an amino acid sequence of SEQ ID NO: 4, containing at least one N-linked glycosylation site, and
wherein the fusion protein, once administered in vivo, results in reduced lysosomal storage.
2. A vector comprising the nucleic acid sequence of claim 1 .
3. A host cell comprising the vector of claim 2 .
4. The host cell of claim 3 , wherein the host cell co-expresses N-acetylglucosamine-1-phosphoTransferase (GNPT).
5. The host cell of claim 4 , wherein the host cell comprises an exogenous nucleic acid encoding GNPT.
6. The host cell of claim 4 , wherein the host cell has activated expression of endogenous GNPT.
7. A method for producing a targeted therapeutic fusion protein, comprising:
a) culturing cells comprising a nucleic acid of claim 1 , wherein the cells co-express N-acetylglucosamine-1-phosphoTransferase (GNPT); and
b) recovering the fusion protein produced by the cells.
8. The nucleic acid of claim 1 , wherein the fusion protein comprises an amino acid sequence at least 90% identical to SEQ ID NO: 18.
9. The nucleic acid of claim 1 , wherein the fusion protein comprises an amino acid sequence at least 90% identical to the amino acid sequence of SEQ ID NO. 23.
10. The nucleic acid of claim 1 , wherein the lysosomal targeting moiety and the lysosomal enzyme is fused via a linker.
11. The nucleic acid of claim 10 , wherein the linker comprises a sequence of
(SEQ ID NO.: 15)
GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAG
GGGGGAP.