TREATMENT OF SANFILIPPO SYNDROME TYPE B
Among other things, the present invention provides methods and compositions of treating Sanfilippo syndrome type B (Sanfilippo B) by, e.g., intrathecal (IT) administration of a Naglu protein. A suitable Naglu protein can be a recombinant, gene-activated or natural protein. In some embodiments, a suitable Naglu protein is a recombinant Naglu protein. In some embodiments, a recombinant Naglu protein is a fusion protein containing a Naglu domain and a lysosomal targeting moiety. In some embodiments, the lysosomal targeting domain is an IGF-II moiety.
1 . A method of treating Sanfilippo syndrome type B (San B) disease comprising a step of
administering intrathecally to a subject in need of treatment a recombinant alpha-N-acetylglucosaminidase (Naglu) protein.
2 . The method of claim 1 , wherein the recombinant Naglu protein is a fusion protein comprising a Naglu domain and a lysosomal targeting moiety.
3 . The method of claim 2 , wherein the Naglu domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:1 (mature human Naglu protein).
4 - 5 . (canceled)
6 . The method of claim 2 , wherein the lysosomal targeting moiety is an IGF-II moiety.
7 . The method of claim 6 , wherein the IGF-II moiety comprises an amino acid sequence at least 70% identical to mature human IGF-II (SEQ ID NO:3).
8 - 9 . (canceled)
10 . The method of claim 6 , wherein the IGF-II moiety comprises an amino acid sequence including residues 8-67 of mature human IGF-II (SEQ ID NO:3).
11 . The method of claim 2 , wherein the fusion protein further comprises a linker between the Naglu domain and the lysosomal targeting moiety.
12 . The method of claim 11 , wherein the linker comprises one or more amino acid sequences of GGGGGAAAAGGGG (SEQ ID NO:4).
13 . The method of claim 12 , wherein the amino acid sequence of GGGGGAAAAGGGG (SEQ ID NO:4) is present in tandem repeats.
14 . The method of claim 13 , wherein the linker further comprises one or more GAP sequences.
15 . The method of claim 14 , wherein the linker comprises amino acid sequence of GAPGGGGGAAAAGGGGGAPGGGGGAAAAGGGGGAPGGGGGAAAAGGGGGA P (SEQ ID NO:5).
16 . The method of claim 2 , wherein the lysosomal targeting moiety is fused directly or via the linker to the C-terminus of the Naglu domain.
17 . The method of claim 2 , wherein the lysosomal targeting moiety is fused directly or via the linker to the N-terminus of the Naglu domain.
18 . The method of claim 1 , wherein the recombinant protein is produced from human cells.
19 . (canceled)
20 . The method of claim 1 , wherein the intrathecal administration results in delivery of the Naglu protein in one or more target brain tissues.
21 . The method of claim 20 , wherein the one or more target brain tissues are selected from the group consisting of tissues from gray matter, white matter, periventricular areas, pia-arachnoid, meninges, neocortex, cerebellum, deep tissues in cerebral cortex, molecular layer, caudate/putamen region, midbrain, deep regions of the pons or medulla, and combinations thereof.
22 . The method of claim 20 , wherein the Naglu protein is delivered to neurons, glial cells, perivascular cells and/or meningeal cells.
23 . The method of claim 1 , wherein the Naglu protein is further delivered to the neurons in the spinal cord.
24 . The method of claim 1 , wherein the intrathecal administration further results in systemic delivery of the Naglu protein in peripheral target tissues.
25 . The method of claim 24 , wherein the peripheral target tissues are selected from liver, kidney, and/or heart.
26 . The method of claim 1 , wherein the intrathecal administration results in lysosomal localization in brain target tissues, spinal cord neurons and/or peripheral target tissues.
27 . The method of claim 1 , wherein the intrathecal administration results in reduction of lysosomal storage in the brain target tissues, spinal cord neurons and/or peripheral target tissues.
28 - 29 . (canceled)
30 . The method of claim 1 , wherein the intrathecal administration results in reduced vacuolization in neurons.
31 . (canceled)
32 . The method of claim 1 , wherein the intrathecal administration results in increased Naglu enzymatic activity in the brain target tissues, spinal cord neurons and/or peripheral target tissues.
33 - 36 . (canceled)
37 . The method of claim 1 , wherein the intrathecal administration results in reduced intensity, severity, or frequency, or delayed onset of at least one symptom or feature of the Sanfilippo B Syndrome.
38 . The method of claim 37 , wherein the at least one symptom or feature of the San B disease is hearing loss, delayed speech development, deficits in motor skills, hyperactivity, mental retardation, aggressiveness and/or sleep disturbances.
39 . The method of claim 1 , wherein the intrathecal administration takes place at an interval selected from once every two weeks, once every month, and once every two months.
40 - 41 . (canceled)
42 . The method of claim 1 , wherein the intrathecal administration is used in conjunction with intravenous administration.
43 - 44 . (canceled)
45 . The method of claim 1 , wherein the intrathecal administration is used in absence of intravenous administration.
46 . method of claim 1 , wherein the intrathecal administration is used in absence of concurrent immunosuppressive therapy.
47 . The method of claim 1 , wherein the Naglu fusion protein is administered at a concentration greater than approximately 20 mg/ml.
48 . A therapeutic fusion protein comprising
a Naglu domain;
a lysosomal targeting moiety, and
wherein, once administered, the therapeutic fusion protein is targeted to lysosomes and is therapeutically active in vivo.
49 . The therapeutic fusion protein of claim 48 , wherein the Naglu domain comprises an amino acid sequence at least 80% identical to SEQ ID NO:1 (mature human Naglu protein).
50 . (canceled)
51 . The therapeutic fusion protein of claim 48 , wherein the lysosomal targeting moiety is an IGF-II moiety.
52 - 53 . (canceled)
54 . The therapeutic fusion protein of claim 48 , wherein the fusion protein further comprises a linker between the Naglu domain and the lysosomal targeting moiety.
55 . The therapeutic fusion protein of claim 54 , wherein the linker comprises amino acid sequence of GAPGGGGGAAAAGGGGGAPGGGGGAAAAGGGGGAPGGGGGAAAAGGGGGA P (SEQ ID NO:5).
56 . The therapeutic fusion protein of claim 54 , wherein the lysosomal targeting moiety is fused directly or via the linker to the C-terminus of the Naglu domain.
57 . A therapeutic fusion protein comprising an amino acid sequence at least 80% identical to SEQ ID NO:6 (the full-length Naglu-IGF-II fusion protein), wherein, once administered, the therapeutic fusion protein is targeted to lysosomes and is therapeutically active in vivo.