Lysosomal storage disease enzymes
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. A method of treating a human patient with lysosomal acid lipase (LAL) deficiency, comprising administering an effective amount of a recombinant human lysosomal acid lipase glycoprotein (rhLAL) to said patient, 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 method of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:2.
3. The method of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:3.
4. The method of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:4.
5. The method of claim 1 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:19.
6. The method of claim 1 , with the further proviso that the N-glycans do not comprise xylose.
7. The method of claim 1 , with the further proviso that the N-glycans do not comprise sialic acid.
8. The method 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 method of claim 8 , wherein the avian cell is an oviduct cell of a chicken.
10. A method of treating a human patient with lysosomal acid lipase (LAL) deficiency, comprising administering an effective amount of a recombinant human lysosomal acid lipase glycoprotein (rhLAL) to said patient, 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 method of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:2.
12. The method of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:3.
13. The method of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:4.
14. The method of claim 10 , wherein the amino acid sequence of said rhLAL is SEQ ID NO:19.
15. The method of claim 10 , with the further proviso that the N-glycans do not comprise xylose.
16. The method of claim 10 , with the further proviso that the N-glycans do not comprise sialic acid.
17. The method 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 method of claim 17 , wherein the avian cell is an oviduct cell of a chicken.
19. The method of claim 1 , comprising administering a pharmaceutical composition consisting essentially of an aqueous solution of
a) 2 mg/mL of rhLAL;
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. The method of claim 10 , comprising administering a pharmaceutical composition consisting essentially of an aqueous solution of
a) 2 mg/mL of rhLAL;
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.