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US-12622952-B2 - Formulations comprising recombinant acid α-glucosidase

US12622952B2US 12622952 B2US12622952 B2US 12622952B2US-12622952-B2

Abstract

Provided are pharmaceutical formulations comprising a recombinant acid α-glucosidase, wherein the recombinant acid α-glucosidase is expressed in Chinese hamster ovary (CHO) cells and comprises an increased content of N-glycan units bearing one or two mannose-6-phosphate residues when compared to a content of N-glycan units bearing one or two mannose-6-phosphate residues of alglucosidase alfa; at least one buffer selected from the group consisting of a citrate, a phosphate and combinations thereof; and at least one excipient selected from the group consisting of mannitol, polysorbate 80, and combinations thereof, wherein the formulation has a pH of from about 5.0 to about 7.0. Also provided are methods of treating Pompe disease using these pharmaceutical formulations.

Inventors

  • Hing Char
  • Sergey Tesler
  • Wendy Sunderland
  • Enrique Diloné
  • Russell Gotschall
  • Hung V. Do

Assignees

  • AMICUS THERAPEUTICS, INC.

Dates

Publication Date
20260512
Application Date
20221107

Claims (20)

  1. 1 . A pharmaceutical formulation comprising: (a) a population of recombinant human acid α-glucosidase (rhGAA) molecules, wherein the rhGAA molecules comprises at least 3.0% of the total N-glycans of the rhGAA are in the form of bis-M6P, and wherein the rhGAA molecules comprise from 2.0 mole to 8.0 mole sialic acid residues per mole rhGAA; (b) at least one buffer selected from a citrate buffer, a phosphate buffer, and combinations thereof; and (c) at least one excipient selected from the group consisting of mannitol, polysorbate 80, and combinations thereof, and wherein the formulation has a pH of from 5.0 to 7.0.
  2. 2 . The pharmaceutical formulation of claim 1 , wherein the rhGAA molecules are present in a concentration of 5 mg/ml to 50 mg/mL.
  3. 3 . The pharmaceutical formulation of claim 2 , wherein the rhGAA molecules are present in a concentration of 15 mg/mL.
  4. 4 . The pharmaceutical formulation of claim 1 , wherein the formulation has a pH of from 5.5 to 7.0.
  5. 5 . The pharmaceutical formulation of claim 4 , wherein the formulation has a pH of 6.0.
  6. 6 . The pharmaceutical formulation of claim 1 , wherein the citrate buffer comprises a potassium, sodium, or ammonium salt.
  7. 7 . The pharmaceutical formulation of claim 1 , wherein the citrate buffer comprises sodium citrate.
  8. 8 . The pharmaceutical formulation of claim 1 , wherein the at least one buffer is present in a concentration of 10 mM to 100 mM.
  9. 9 . The pharmaceutical formulation of claim 8 , wherein the at least one buffer is present in a concentration of 25 mM.
  10. 10 . The pharmaceutical formulation of claim 1 , wherein trehalose, sucrose, glycine, or combinations thereof is excluded.
  11. 11 . The pharmaceutical formulation of claim 1 , wherein the at least one excipient is mannitol present in a concentration of 10 mg/ml to 50 mg/mL.
  12. 12 . The pharmaceutical formulation of claim 1 , wherein the at least one excipient is polysorbate 80 present in a concentration of 0.2 mg/ml to 0.5 mg/mL.
  13. 13 . The pharmaceutical formulation of claim 1 , wherein the at least one excipient comprises mannitol and polysorbate 80, and wherein the mannitol is present at a concentration of 20 mg/mL and the polysorbate 80 is present at a concentration of 0.5 mg/mL.
  14. 14 . The pharmaceutical formulation of claim 1 , further comprising: (a) an alkalizing agent; and/or (b) an acidifying agent, wherein the alkalizing agent and/or acidifying agent are present in amounts to maintain the pharmaceutical formulation at a pH of from 5.0 to 6.0.
  15. 15 . The pharmaceutical formulation of claim 1 , wherein at least 75% of the rhGAA molecules comprise a glycan bearing bis-M6P at the first potential N-glycosylation site.
  16. 16 . The pharmaceutical formulation of claim 1 , wherein the rhGAA molecules comprise from 3.0 mole to 5.0 mole mannose-6-phosphate (M6P) residues per mole rhGAA.
  17. 17 . The pharmaceutical formulation of claim 1 , wherein the rhGAA molecules comprise from 4.0 mole to 7.0 mole sialic acid residues per mole rhGAA.
  18. 18 . The pharmaceutical formulation of claim 1 , wherein the rhGAA molecules comprise at least 3 mole M6P residues per mole of rhGAA, and at least 4.0 mole sialic acid residues per mole rhGAA.
  19. 19 . The pharmaceutical formulation of claim 1 , wherein 75% to 80% of the rhGAA molecules comprise a glycan bearing bis-M6P at the first potential N-glycosylation site, 40% to 60% of the rhGAA molecules comprise a glycan bearing mono-mannose-6-phosphate (mono-M6P) at the second potential N-glycosylation site, 40% to 60% of the rhGAA molecules comprise a glycan bearing bis-M6P at the fourth potential N-glycosylation site, and 25% to 40% of the rhGAA molecules comprise a glycan bearing mono-M6P at the fourth potential N-glycosylation site.
  20. 20 . The pharmaceutical formulation of claim 1 , wherein 20% of the rhGAA molecules comprise a sialic residue at the third potential N-glycosylation site, 70% of the rhGAA molecules comprise a sialic residue at the fifth potential N-glycosylation site, and 80% of the rhGAA molecules comprise a sialic acid residue at the sixth potential N-glycosylation site.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application is a divisional of U.S. application Ser. No. 16/654,521, filed Oct. 16, 2019, which is a continuation of U.S. application Ser. No. 15/473,999, filed Mar. 30, 2017, now U.S. Pat. No. 10,512,676, which claims the benefit under 35 U.S.C. § 119 (e) to U.S. Provisional Application No. 62/315,436, filed Mar. 30, 2016, and U.S. Provisional Application No. 62/457,588, filed Feb. 10, 2017, the entire contents of which are incorporated herein by reference in their entirety. TECHNICAL FIELD Principles and embodiments of the present invention relate generally to formulations comprising recombinant acid α-Glucosidase, and particularly liquid formulations. BACKGROUND Pompe disease, also known as acid maltase deficiency or glycogen storage disease type II, is one of several lysosomal storage disorders. Lysosomal storage disorders are a group of autosomal recessive genetic diseases characterized by the accumulation of cellular glycosphingolipids, glycogen, or mucopolysaccharides within intracellular compartments called lysosomes. Individuals with these diseases carry mutant genes coding for enzymes which are defective in catalyzing the hydrolysis of one or more of these substances, which then build up in the lysosomes. Other examples of lysosomal disorders include Gaucher disease, GM1-gangliosidosis, fucosidosis, mucopolysaccharidoses, Hurler-Scheie disease, Niemann-Pick A and B diseases, and Fabry disease. Pompe disease is also classified as a neuromuscular disease or a metabolic myopathy. Pompe disease is estimated to occur in about 1 in 40,000 births, and is caused by a mutation in the GAA gene, which codes for the enzyme lysosomal α-glucosidase (EC:3.2.1.20), also commonly known as acid α-glucosidase. Acid α-glucosidase is involved in the metabolism of glycogen, a branched polysaccharide which is the major storage form of glucose in animals, by catalyzing its hydrolysis into glucose within the lysosomes. Because individuals with Pompe disease produce mutant, defective acid α-glucosidase which is inactive or has reduced activity, glycogen breakdown occurs slowly or not at all, and glycogen accumulates in the lysosomes of various tissues, particularly in striated muscles, leading to a broad spectrum of clinical manifestations, including progressive muscle weakness and respiratory insufficiency. Tissues such as the heart and skeletal muscles are particularly affected. Pompe disease can vary widely in the degree of enzyme deficiency, severity and age of onset, and over 500 different mutations in the GAA gene have been identified, many of which cause disease symptoms of varying severity. The disease has been classified into broad types: early onset or infantile and late onset. Earlier onset of disease and lower enzymatic activity are generally associated with a more severe clinical course. Infantile Pompe disease is the most severe, resulting from complete or near complete acid α-glucosidase deficiency, and presents with symptoms that include severe lack of muscle tone, weakness, enlarged liver and heart, and cardiomyopathy. The tongue may become enlarged and protrude, and swallowing may become difficult. Most affected children die from respiratory or cardiac complications before the age of two. Late onset Pompe disease can present at any age older than 12 months and is characterized by a lack of cardiac involvement and better short-term prognosis. Symptoms are related to progressive skeletal muscle dysfunction, and involve generalized muscle weakness and wasting of respiratory muscles in the trunk, proximal lower limbs, and diaphragm. Some adult patients are devoid of major symptoms or motor limitations. Prognosis generally depends on the extent of respiratory muscle involvement. Most subjects with Pompe disease eventually progress to physical debilitation requiring the use of a wheelchair and assisted ventilation, with premature death often occurring due to respiratory failure. Recent treatment options for Pompe disease include enzyme replacement therapy (ERT) with recombinant human acid α-glucosidase (rhGAA). Conventional rhGAA products are known under the names alglucosidase alfa, Myozyme® or Lumizyme® from Genzyme, Inc. ERT is a chronic treatment required throughout the lifetime of the patient, and involves administering the replacement enzyme by intravenous infusion. The replacement enzyme is then transported in the circulation and enters lysosomes within cells, where it acts to break down the accumulated glycogen, compensating for the deficient activity of the endogenous defective mutant enzyme, and thus relieving the disease symptoms. The way in which replacement enzymes, such as rhGAA, are prepared, stored, transported and administered to patients is difficult. The enzymes used in ERT are generally relatively complex and delicate, making selection of accompanying buffers, excipients, etc. critical. If the enzyme is not preserved properly, then high quantities