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Claims for Patent: 5,549,892

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Claims for Patent: 5,549,892

Title: Enhanced in vivo uptake of glucocerebrosidase
Abstract:A pharmaceutical composition comprising remodelled recombinant glucocerebrosidase (GCR) is described that provides a therapeutic effect at doses that are lower then those required using remodelled naturally occurring GCR. A method of treating patients with Gaucher's disease using remodelled recombinant GCR is also provided. In vivo uptake of exogenous molecules can be determined by extracting a mixture of cells from a subject, enriching the target cells in vitro, lysing the cells and determining the amount of exogenous molecules.
Inventor(s): Friedman; BethAnn (Arlington, MA), Hayes; Michael (Acton, MA)
Assignee: Genzyme Corporation (Cambridge, MA)
Application Number:08/080,855
Patent Claims: 1. A pharmaceutical composition, comprising:

remodeled recombinant GCR, obtained from CHO cells, wherein the remodeled recombinant GCR has exposed mannose terminal residues on appended oligosaccharides, in an effective dosage suitable for significantly alleviating clinical symptoms of Gaucher's disease, such that the dose is substantially less than the effective dose using remodeled naturally occurring GCR.

2. The pharmaceutical composition of claim 1, wherein the recombinant GCR differs from naturally occurring GCR by having a histidine at amino acid number 495.

3. The pharmaceutical composition of claim 1, wherein the remodelled recombinant GCR has increased fucose compared to remodelled naturally occurring GCR.

4. The pharmaceutical composition of claim 1 wherein the remodelled recombinant GCR has increased N-acetyl glucosamine residues compared to remodelled naturally occurring GCR.

5. A method for treating a human subject having Gaucher's disease using exogenous GCR, comprising:

(a) providing a recombinant form of GCR obtained from CHO cells and capable of effectively targeting cells abnormally deficient in GCR, wherein the recombinant GCR has exposed terminal mannose residues on appended oligosaccharides;

(b) administering such form of the GCR to the subject in doses sufficient to achieve a therapeutic effect, each dose being dependent on the effective targeting of cells abnormally deficient in GCR; and each dose of such form of GCR being substantially less than the dose of naturally occurring GCR that would otherwise be administered in a similar manner to achieve the therapeutic effect.

6. A method according to claim 5, wherein the recombinant GCR differs from naturally occurring GCR by having a histidine at amino acid number 495.

7. A method according to claim 5, wherein the recombinant GCR has a carbohydrate moiety having increased fucose and N-acetylglucosamine residues compared to the naturally occurring forms of GCR.

8. A method according to claim 5, wherein the targeting capability in step (a) is determined in relation to the uptake by a population of target cells and the recombinant GCR has increased affinity for the target cells in comparison with that of naturally occurring GCR.

9. A method according to claim 8, wherein the cells at the target site are Kupffer cells in the liver.

10. A pharmaceutical composition for significantly alleviating clinical symptoms of Gaucher's disease comprising remodeled recombinant GCR obtained from CHO cells the remodeled recombinant GCR having an effective dosage that is substantially less than the effective dosage for remodeled naturally occurring GCR.
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