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 targeted therapeutic comprising:
a lysosomal enzyme; and
a lysosomal targeting moiety, wherein the lysosomal targeting moiety is a peptide containing at least one N-linked glycosylation site.
2 . (canceled)
3 . The targeted therapeutic of claim 1 , wherein the peptide contains N-linked glycosylation containing a M6P group.
4 . (canceled)
5 . The targeted therapeutic of claim 1 , wherein the lysosomal enzyme is an N-Acetylglucosaminidase (Naglu) protein.
6 . The targeted therapeutic of claim 1 , wherein the Naglu protein comprises an amino acid sequence at least 80% identical to SEQ ID NO.: 1.
7 .- 9 . (canceled)
10 . The targeted therapeutic of claim 1 , wherein the lysosomal targeting moiety is a peptide selected from the group consisting of prosaposin, cellular repressor of E1A-stimulate genes, leukemia inhibitory factor, and fragments thereof.
11 .- 12 . (canceled)
13 . The targeted therapeutic of claim 10 , wherein the peptide comprises a sequence at least 95% identical to SEQ ID NO. 4, 6, or 9.
14 .- 17 . (canceled)
18 . The targeted therapeutic of claim 1 , wherein the lysosomal targeting moiety and the lysosomal enzyme is fused via a linker.
19 . The targeted therapeutic of claim 1 , wherein the linker comprises a sequence of
(SEQ ID NO.: 15)
GAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAGGGGGGAPGGGGGAAAAAG
GGGGGAP.
20 . (canceled)
21 . The targeted therapeutic of claim 1 , wherein the fusion protein is at least 95% identical to the amino acid sequence of SEQ ID NO. 18, 19 or 20.
22 .- 24 . (canceled)
25 . The targeted therapeutic of claim 1 , wherein the fusion protein comprises a sequence at least 95% identical to the amino acid sequence of SEQ ID NO. 23, 24 or 25.
26 . (canceled)
27 . A nucleic acid encoding the targeted therapeutic fusion protein of claim 1 .
28 . A vector comprising the nucleic acid sequence of claim 27 .
29 . A host cell comprising the vector of claim 28 .
30 .- 33 . (canceled)
34 . The host cell of claim 29 , wherein the host cell co-expresses N-acetylglucosamine-1-phosphoTransferase (GNPT).
35 . The host cell of claim 34 , wherein the host cell comprises an exogenous nucleic acid encoding GNPT.
36 . The host cell of claim 34 , wherein the host cell has activated expression of endogenous GNPT.
37 . A method for producing a targeted therapeutic fusion protein, comprising:
a) culturing cells comprising a nucleic acid encoding a fusion protein of claim 1 , wherein the cells co-express N-acetylglucosamine-1-phosphoTransferase (GNPT); and
b) recovering the fusion protein produced by the cells.
38 . A pharmaceutical composition comprising the targeted therapeutic of claim 1 , and a pharmaceutical acceptable carrier.
39 . A method of treating a lysosomal storage disease comprising administering to a subject in need of treatment a pharmaceutical composition of claim 38 .
40 . The method of claim 39 , wherein the lysosomal storage disease is Sanfilippo syndrome type B.
41 . (canceled)