NZ784003B2 - Augmented acid alpha-glucosidase for the treatment of pompe disease - Google Patents

Abstract

method for treating Pompe disease including administration of recombinant human acid a-glucosidase having optimal glycosylation with mannose-6-phosphate residues in combination with an amount of miglustat effective to maximize tissue uptake of recombinant human acid a-glucosidase while minimizing inhibition of the enzymatic activity of the recombinant human acid a-glucosidase is provided.

Claims (4)

1. A kit comprising a first composition comprising recombinant human acid a-glucosidase (rhGAA) molecules and a second composition comprising miglustat, wherein the first composition comprises 5 mg/kg to 20 mg/kg rhGAA, wherein the second composition comprises 50 mg to 600 mg miglustat, and wherein at least 3% of the total glycans on the rhGAA molecules are bis-M6P.

2. A kit sing a first composition comprising recombinant human acid a-glucosidase (rhGAA) molecules and a second ition comprising miglustat, n the first composition comprises 5 mg/kg to 20 mg/kg rhGAA, wherein the second composition ses 50 mg to 600 mg miglustat, and wherein 40%-60% of the ans on the rhGAA molecules are complex type ans.

3. The kit according to claim 1 or claim 2, wherein the rhGAA molecules are expressed in Chinese hamster ovary (CHO) cells. 4. The kit according to any one of claims 1-3, wherein 30% to 99% of the rhGAA molecules comprise one or more N-glycan units bearing one or two mannosephosphate residues. 5. The kit according to any one of claims 1-4, wherein the rhGAA molecules comprise on average from 0.5 to 7.0 moles of an units bearing one or two mannosephosphate residues per mol of rhGAA molecules. 6. The kit according to any one of claims 1-5, wherein the rhGAA molecules comprise on average 2.0 to 8.0 moles of sialic acid residues per mol of rhGAA molecules. 7. The kit according to any one of claims 1-6, wherein (a) 50% to 95% of the rhGAA molecules comprise an N-glycan unit bearing bis-M6P at the first N-glycosylation site, (b) 40% to 60% of the rhGAA molecules comprise an N-glycan unit bearing mono-M6P at the second N- glycosylation site; (c) 40% to 60% of the rhGAA molecules comprise an N-glycan unit bearing bis-M6P at the fourth N-glycosylation site; and/or (d) 20% to 40% of the rhGAA molecules comprise an N-glycan unit bearing mono-M6P at the fourth N-glycosylation site. 8. The kit according to any one of claims 1-7, wherein the first composition comprises 5 mg/kg to 20 mg/kg rhGAA molecules, and the second composition comprises 233 mg to 500 mg miglustat; wherein the first composition comprises 20 mg/kg rhGAA molecules and the second composition comprises 260 mg miglustat; or wherein the first composition comprises 10 mg/kg to 20 mg/kg rhGAA molecules. 9. The kit according to any one of claims 1-8, wherein the rhGAA molecules have a shorter halflife than alglucosidase alfa in ; wherein the half-life of the rhGAA molecules is 20% to 30% shorter than alglucosidase alfa in ; or wherein the half-life of the rhGAA molecules is 25% r than alglucosidase alfa in plasma. 10. The kit according to any one of claims 1-9, wherein the rhGAA molecules have on average 1.2 more moles of N-glycan units g two mannosephosphate residues per compared to alglucosidase alfa. 11. The kit according to any one of claims 1-10, wherein the rhGAA molecules induce a lower incidence of anti-drug antibodies than alglucosidase alfa; wherein the rhGAA molecules reduce glycogen in muscle s more effectively than alglucosidase alfa; wherein the rhGAA molecules reduce vacuoles in muscle fibers more ively than alglucosidase alfa; wherein the rhGAA molecules clear lysosomal glycogen more effectively than alglucosidase alfa; wherein the rhGAA molecules increase muscle function more efficiently than alglucosidase alfa; wherein the rhGAA molecules internalize into muscle lasts more ently than alglucosidase alfa; and/or wherein the rhGAA molecules reduce lysosomal proliferation more ently than alglucosidase alfa. 12. The kit according to any one of claims 1-11, wherein the rhGAA molecules bind cationindependent mannosephosphate receptor to a greater degree than alglucosidase alfa; or wherein 43% more of the rhGAA molecules bind cation-independent mannosephosphate receptor than osidase alfa. 13. The kit according to any one of claims 1-12, n the rhGAA molecules comprise on average at least 1 mol bis-M6P per mol rhGAA les; or wherein the rhGAA molecules comprise on average 1.3 mol bis-M6P per mol rhGAA molecules. 14. The kit according to any one of claims 2-13, wherein at least 3% of the total glycans on the rhGAA molecules are bis-M6P, and on average less than 25% of the total rhGAA contain no phosphorylated glycan binding to CIMPR; n at least 17% of the total glycans on the rhGAA molecules are bis-M6P; wherein 3% to 25% of the total glycans on the rhGAA molecules are bis-M6P; or wherein 17% to 25% of the total glycans on the rhGAA les are bis-M6P. 15. The kit according to any one of claims 1-14, wherein the kit comprises 200 mg to 600 mg tat; wherein the kit comprises 65 mg miglustat; wherein the kit comprises 130 mg miglustat; wherein the kit comprises 195 mg miglustat; or wherein the kit comprises 260 mg miglustat. 16. Use of recombinant human acid a-glucosidase (rhGAA) les and miglustat in the preparation of a medicament for the treatment of Pompe disease, wherein the medicament comprises 5 mg/kg to 20 mg/kg rhGAA and 50 mg to 600 mg miglustat, wherein at least 3% of the total glycans on the rhGAA molecules are bis-M6P. 17. Use of recombinant human acid a-glucosidase (rhGAA) molecules and miglustat in the preparation of a medicament for the treatment of Pompe disease, wherein the medicament comprises 5 mg/kg to 20 mg/kg rhGAA and 50 mg to 600 mg miglustat, n 40%-60% of the N-glycans on the rhGAA molecules are complex type N-glycans. 18. The use according to claim 16 or 17, wherein the medicament is for the treatment of Pompe disease in an enzyme replacement therapy experienced patient. 19. The use according to any one of claims 16-18, wherein the rhGAA molecules are expressed in Chinese hamster ovary (CHO) cells. 20. The use according to any one of claims 16-19, wherein 30% to 99% of the rhGAA molecules comprise one or more N-glycan units bearing one or two mannosephosphate residues. 21. The use ing to any one of claims 16-20, wherein the rhGAA molecules se on average from 0.5 to 7.0 moles of N-glycan units bearing one or two mannosephosphate residues per mol of rhGAA molecules. 22. The use according to any one of claims 16-21, n the rhGAA molecules comprise on average 2.0 to 8.0 moles of sialic acid residues per mol of rhGAA molecules. 23. The use according to any one of claims 16-22, n (a) 50% to 95% of the rhGAA les comprise an N-glycan unit g bis-M6P at the first N-glycosylation site, (b) 40% to 60% of the rhGAA les comprise an N-glycan unit g 6P at the second N-glycosylation site; (c) 40% to 60% of the rhGAA molecules comprise an N-glycan unit bearing bis-M6P at the fourth N-glycosylation site; and/or (d) 20% to 40% of the rhGAA molecules comprise an N-glycan unit bearing mono-M6P at the fourth N-glycosylation site. 24. The use according to any one of claims 16-23, wherein the first composition comprises 5 mg/kg to 20 mg/kg rhGAA molecules, and the second composition comprises 233 mg to 500 mg miglustat; wherein the first composition comprises 20 mg/kg rhGAA molecules and the second composition comprises 260 mg miglustat; or wherein the first composition comprises 10 mg/kg to 20 mg/kg rhGAA molecules. 25. The use according to any one of claims 16-24, wherein the rhGAA molecules have a shorter half-life than alglucosidase alfa in plasma; wherein the half-life of the rhGAA molecules is 20% to 30% shorter than alglucosidase alfa in plasma; or wherein the half-life of the rhGAA molecules is 25% shorter than alglucosidase alfa in plasma. 26. The use ing to any one of claims 16-25, wherein the rhGAA molecules have on average 1.2 more moles of N-glycan units bearing two mannosephosphate residues per compared to alglucosidase alfa. 27. The use according to any one of claims 16-26, n the rhGAA molecules induce a lower incidence of anti-drug antibodies than alglucosidase alfa; wherein the rhGAA molecules reduce glycogen in muscle tissues more effectively than alglucosidase alfa; wherein the rhGAA molecules reduce vacuoles in muscle fibers more effectively than alglucosidase alfa; wherein the rhGAA molecules clear lysosomal glycogen more effectively than alglucosidase alfa; wherein the rhGAA molecules increase muscle function more efficiently than osidase alfa; wherein the rhGAA molecules internalize into muscle fibroblasts more efficiently than alglucosidase alfa; and/or wherein the rhGAA molecules reduce lysosomal proliferation more efficiently than osidase alfa. 28. The use according to any one of claims 16-27, wherein the rhGAA molecules bind cationindependent mannosephosphate or to a greater degree than alglucosidase alfa, or n 43% more of the rhGAA molecules bind cation-independent mannosephosphate or than alglucosidase alfa. 29. The use according to any one of claims 16-28, n the rhGAA molecules comprise on average 1 mol bis-M6P per mol rhGAA molecules; wherein the rhGAA molecules comprise on average 1.3 mol P per mol rhGAA molecules. 30. The use according to any one of claims 16-29, wherein at least 3% of the total glycans on the rhGAA molecules are bis-M6P, and on average less than 25% of the total rhGAA n no phosphorylated glycan binding to CIMPR; wherein at least 17% of the total glycans on the rhGAA molecules are bis-M6P; wherein 3% to 25% of the total glycans on the rhGAA molecules are bis-M6P; or wherein 17% to 25% of the total glycans on the rhGAA molecules are bis-M6P. 31. The use according to any one of claims 16-30, wherein the medicament comprises 200 mg to 600 mg miglustat; wherein the medicament ses 65 mg miglustat; wherein the medicament comprises 130 mg miglustat; wherein the medicament comprises 195 mg miglustat; or wherein the medicament comprises 260 mg miglustat. ATBZOOiMiglustat Protein Unfolder " ,- — pH 7.4 (pH of Blood) 0')O ... pH 74 + 10p,M Miglustat _._ pH 74 + 30 FM Miglustat Percent _.._ pH 74 +100pM Miglustat — pH 5.2 (pH of Lysozyme) “10,000 g 6 E, 8,000 5 g 6,000 4 E g. 30. g 4,000 2% €15) 2,000 1 0 0 Myozyme 15,000 5 e 4 $10,000 § 3‘ e: 3 E. g n. e 5,000 ED 1 0 0 rhGAA lacks M6P; rhGAA contains M6P; cannot be targeted targeted to lysosomes to iysosomes FIG. ZB ATBZOO Vector Construction 9 Host Ceii: DG44 0 Vector: pOptivec-110 I3 invitrogen vector [3952 aa, native signal TK polyA peptide

4. Screening Pressure: -HT, SOOnM MTX EMCV-IRES Lumizyme «10,000 g 6 E. 8,000 5 .23 6,000 4 E e 3 4.. :3, 4,000 2% 51; 2,000 1 0 0 5,000 (F460) 4,000 'I I I I I I I .I .038C) [mM] activity 2,000 M6P GAA 1,000 0: Fo—xmw-hcn'm" rhGAA lacks M6P; rhGAA contains M6P; cannot be targeted targeted to mes to lysosomes Embodiment 1 5,000 60) .b OOO (F4 (.10O o\° 'I I I I I I I I L01 activi$2000 7 NOD M6P[mM] 1,000 —