Lysosomal storage disease enzymes
View Patent ↗The present invention provides compositions of recombinant human lysosomal acid lipase having particular glycosylation patterns for internalization into target cells, a vector containing the nucleic acid encoding human lysosomal acid lipase, a host cell transformed with the vector, pharmaceutical compositions comprising the recombinant human lysosomal acid lipase and method of treating conditions associated with lysosomal acid lipase deficiency.
1. An isolated recombinant human lysosomal acid lipase glycoprotein (rhLAL), wherein the rhLAL consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:19, with N-glycans bonded to the asparagine residues (Asn) that correspond to Asn 36 , Asn 101 , Asn 161 , Asn 273 and Asn 321 of SEQ ID NO:1, with the proviso that the N-glycans do not comprise fucose, and the proviso that the rhLAL does not contain O-glycans.
2. The rhLAL of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:2.
3. The rhLAL of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:3.
4. The rhLAL of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:4.
5. The rhLAL of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:19.
6. The rhLAL of claim 1 , with the further proviso that the N-glycans do not comprise xylose.
7. The rhLAL of claim 1 , with the further proviso that the N-glycans do not comprise sialic acid.
8. The rhLAL of claim 1 , wherein the rhLAL is produced by a process that comprises expressing a nucleic acid that encodes said amino acid sequence in an avian cell.
9. The rhLAL of claim 8 , wherein the avian cell is an oviduct cell of a chicken.
10. An isolated recombinant human lysosomal acid lipase glycoprotein (rhLAL), wherein the rhLAL consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ ID NO:19, with N-glycans bonded to the asparagine residues (Asn) that correspond to Asn 36 , Asn 101 , Asn 161 , Asn 273 and Asn 321 of SEQ ID NO:1, with the proviso that the N-glycans do not comprise fucose, and the proviso that the rhLAL does not contain O-glycans, and wherein the N-glycans bonded to the Asn residues that correspond to Asn 36 , Asn 101 , Asn 161 , Asn 273 and Asn 321 of SEQ ID NO:1 are:
a) at Asn 36 , GlcNAc4Man3GlcNAc2, or
Gal1GlcNAc4Man3GlcNAc2;
b) at Asn 101 , Phos2Man7GlcNAc2;
c) at Asn 161 , Phos1Man6GlcNAc2,
GlcNAc1Phos1Man6GlcNAc2;
Man3GlcNAc2;
GlcNAc2Man3GlcNAc2;
GlcNAc3Man3GlcNAc2;
GlcNAc4Man3GlcNAc2, or
Gal1GlcNAc4Man3GlcNAc2;
d) at Asn 273 , Man7GlcNAc2,
Man8GlcNAc2,
Man9GlcNAc2,
Phos1Man8GlcNAc2, or
Phos1Man9GlcNAc2; and
e) at Asn 321 , GlcNAc2Man3GlcNAc2,
GlcNAc3Man3GlcNAc2,
GlcNAc4Man3GlcNAc2,
Gal1GlcNAc4Man3GlcNAc2,
GlcNAc5Man3GlcNAc2,
Gal1GlcNAc5Man3GlcNAc2,
GlcNAc6Man3GlcNAc2, or
Gal1GlcNAc6Man3GlcNAc2; wherein
Man=mannose,
GlcNAc=N-Acetyl Glucosamine,
Phos=phosphate, and
Gal=galactose.
11. The rhLAL of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:2.
12. The rhLAL of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:3.
13. The rhLAL of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:4.
14. The rhLAL of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:19.
15. The rhLAL of claim 10 , with the further proviso that the N-glycans do not comprise xylose.
16. The rhLAL of claim 10 , with the further proviso that the N-glycans do not comprise sialic acid.
17. The rhLAL of claim 10 , wherein the rhLAL is produced by a process that comprises expressing a nucleic acid that encodes said amino acid sequence in an avian cell.
18. The rhLAL of claim 17 , wherein the avian cell is an oviduct cell of a chicken.
19. A pharmaceutical composition consisting essentially of an aqueous solution of
a) 2 mg/mL of hLAL of claim 1 ;
b) 13.7 mg/mL of trisodium citrate dehydrate;
c) 1.57 mg/mL of citric acid monohydrate; and
d) 10 mg/mL of human serum albumin; wherein the pH of the pharmaceutical composition is 5.9±0.2.
20. A pharmaceutical composition consisting essentially of an aqueous solution of
a) 2 mg/mL of hLAL of claim 10 ;
b) 13.7 mg/mL of trisodium citrate dehydrate;
c) 1.57 mg/mL of citric acid monohydrate; and
d) 10 mg/mL of human serum albumin; wherein the pH of the pharmaceutical composition is 5.9±0.2.