galsulfase - Naglazyme; N-acetylgalactosamine 4-sulfatase, recombinant; Aryplase; chondroitinase; rhASB
Status: approved and marketed in U.S. and EU
Organizations involved:
BioMarin Pharmaceutical Inc. – Manuf.; R&D; Tech.; World mark.
Hollister-Stier Labs. – Manuf. other
Jubilant Organosys Ltd. – Parent
AnGes MG – Japan mark.
CSL, Inc. – Tech.
Tektagen Inc. – Tech.
Cross ref.: See also the entry for alpha-L-iduronidase, another enzyme from BioMarin for treatment of deficiency of a different GAG-degrading enzyme (Hurler syndrome).
Description: Naglazyme is an aqueous formulation of galsulfase or recombinant human N-acetylgalactosamine 4-sulfatase (arylsulfatase B; ASB) glycosylated protein expressed by the transformed Chinese hamster ovary (CHO) CSL4S-342 cell line. ASB is a lysosomal hydrolase enzyme that catalyzes the cleavage of the sulfate ester from terminal N–acetylgalactosamine 4-sulfate residues of the glycosaminoglycans (GAG) chondroitin 4-sulfate and dermatan sulfate.
The glycoprotein is comprised of 495 amino acids with six asparagine-linked glycosylation sites, four of which carry a bis mannose-6-phosphate mannose7 oligosaccharide needed for specific cellular recognition. The predicted molecular weight of the mature enzyme is about 55.87 kDa, with a predicted molecular formula of C2529H3843N689O716S16. The predicted protein sequence contains six potential N-linked oligosaccharide modification sites of which four may be used. The six asparagine-linked glycosylation sites carry a mixture of complex, high-mannose and phosphorylated high-mannose oligosaccharides. A mature form of the intracellular protein has three peptides attached by cystine bonds. The largest peptide has a molecular weight of 47 kDa; the other two have a molecular weight of 6 and 7 kDa, respectively. The cysteine residue in position 53 is post-translationally modified to formylglycine within the endoplasmic reticulum of human cells. This modification, common to all sulfatases, is required for enzymatic activity.
Galsulfase has a specific activity of approximately 70 U/mg protein content. One activity unit (U) is defined as the amount of enzyme required to convert 1 µmole of 4-methylumbelliferyl sulfate to 4-methylumbelliferone and free sulfate per minute at 37°C.
Naglazyme is packaged in vials (type I borosilicate glass) with a stopper (siliconized chlorobutyl rubber) and a seal (aluminium) with a flipoff cap (polypropylene). For intravenous infusion, Naglazyme is diluted in 0.9% Sodium Chloride Injection, USP. The solution in each vial contains a nominal galsulfase concentration of 1 mg/mL (expressed as protein concentration) at a pH of approximately 5.8. The extractable volume of 5 mL from each vial provides 5 mg galsulfase, 43.8 mg sodium chloride, 6.20 mg sodium phosphate monobasic monohydrate, 1.34 mg sodium phosphate dibasic heptahydrate, and 0.25 mg polysorbate 80 (Tween 80). Naglazyme does not contain preservatives, and vials are for single use only. The shelf life (dating period) is 12 months stored at 1-8˚C (refrigerated).
Nomenclature: Arylsulfatase B, rDNA [BIO]; Naglazyme [TR]; Aryplase [TR former]; galsulfase [USAN INN]; sulfatase, acetylgalactosamine 4- (human CSL4S-342 cell) [CAS]; N-acetylgalactosamine 4-sulfatase (human CSL4S-342 cell) [CAS]; 552858-79-4 [CAS RN]; chondroitinase [SY]; chondroitinsulfatase [SY]; ASB [SY]; G4S [SY]; MPS6 [SY]; 4-sulfatase [SY]; N-acetylgalactosamine 4-sulfatase [SY]; chondroitin 4-sulfatase [SY]; nitrocatechol sulfatase [SY]; rhASB [SY]; BM-102 [SY]; EC 3.1.6.12 [EC]
Biological.: Galsulfase is an enzyme that catalyzes the hydrolysis of the 4-sulfate groups of certain glycosaminoglycans (GAGs), particularly the N-acetyl-D-galactosamine 4-sulfate units of chondroitin sulfate and dermatan sulfate. Galsulfase is used for enzyme replacement therapy for treatment of mucopolysaccharidosis VI (MPS VI; Maroteaux-Lamy syndrome; polydystrophic dwarfism). Galsulfase uptake by cells into lysosomes is most likely mediated by the binding of its bisphosphorylated oligomannose7 oligosaccharide chains to specific mannose-6-phosphate receptors.
Glycosaminoglycans (GAGs) are the most abundant heteropolysaccharides in the human body. GAGs are located primarily on the surface of cells or in the extracellular matrix (ECM), and occur in tissues including cartilage, bone and heart valves. GAGs include hyaluronic acid, dermatan sulfate, chondroitin sulfate, heparin, heparan sulfate, and keratan sulfate. GAGs are long unbranched polysaccharides containing a repeating disaccharide unit, each composed of either N-acetylgalactosamine (GalNAc) or N-acetylglucosamine (GlcNAc) and a uronic acid, e.g., glucuronate or iduronate. Chondroitin is the most abundant GAG. Chondroitin 4- and 6-sulfates are composed of D-glucuronate plus GalNAc-4- or 6-sulfate with a beta(1, 3) linkage. GAGs are highly negatively charged molecules. Their extended conformation imparts high viscosity in aqueous solutions and low compressibility, making these molecules ideal for use as a lubricating fluid in the joints (e.g., see the Hyaluronic Acid product entries), while their rigidity provides structural integrity to cells and provides passageways between cells, allowing for cell migration.
The turnover (catabolism) of GAGs depends on their cellular internalization by endocytosis, delivery to the lysosomes, and digestion by lysosomal enzymes. GAG enzyme deficiencies lead to the accumulation of mucopolysaccharides in the lysosomes of the cells in the connective tissue and to an increase in their excretion in the urine. Thus, these diseases are referred to as lysosomal storage diseases.
MPS VI (mucopolysaccharidosis VI; Maroteaux-Lamy syndrome) is an inherited (genetic), debilitating, life-threatening lysosomal storage disease for which no drug therapies are currently available. [See the Medical section below]. MPS VI is caused by the deficiency of functional arylsulfatase B. Over 45 mutations have been associated with deficiencies in functional enzyme activity. Enzyme deficiency leads to the accumulation of GAGs in the lysosomes of cells, giving rise to progressive cellular, tissue, and organ system dysfunction.
Arylsulfatase B precursor (and mature enzyme) cellular uptake is mediated by a high affinity mannose-6-phosphate receptor expressed on most, if not all, cells. Once bound to this receptor, the enzyme is endocytosed through coated pits and transported to the lysosomes. Inside cells, proteases remove the leader sequence of the precursor form, leaving mature, active enzyme. At the pH of lysosomes, the enzyme is active and begins removing sulfate residues from accumulated certain glycosaminoglycans, e.g., dermatan sulfate. In MPS VI fibroblasts, the clearance of stored dermatan sulfate is rapid and easily demonstrated within 92 hours of enzyme exposure. Naglazyme is designed to address the underlying cause of MPS VI, i.e., arylsulfatase B deficiency, and provide the sufficient levels of the enzyme in MPS VI patients.
Deficiency of functional arylsulfatase B has also been linked to cystic fibrosis (CF). The mucus that excessively accumulates in the lungs of patients with CF may be linked to the deficiency of the enzyme, as reported in July 2003 (Chest. 2003;123:2130-2139). Arylsulfatase B deficiency in CF cell lines was first reported by Neufeld, et., NIH, in 1970. The Pseudomonas bacteria that often infect people with CF may utilize the GAGs that are not metabolized because of insufficient arylsulfatase B, with the excess GAGs perhaps helping the bacteria flourish.
Companies.: BioMarin Pharmaceutical Inc. (CBER/FDA est. no. 1649) developed, manufactures and exclusively markets Naglazyme. Filling, finishing and packaging are performed by Hollister-Stier Labs., which was acquired by Jubilant Organosys Ltd. in April 2007.
In Jan. 2007, BioMarin granted AnGes MG exclusive rights to market Naglazyme in Japan for MPS-VI.
In Nov. 2011, BioMarin received FDA approval for expanded manufacturing facilities in Novato, CA, including manufacture of Naglazyme.
In Dec. 2011, the Committee for Medicinal Products for Human Use (CHMP), European Medicines Agency (EMA), recommended approval of the new manufacturing facility expansion in Novato, CA. Final approval by the EMA was expected in the first quarter of 2012, which would allow for products manufactured at the facility to be distributed in the European Union.
Manufacture: BioMarin manufactures Aryplase in its own Galli Drive GMP facilities, which have been approved and used for manufacture of its currently marketed enzyme replacement product, alpha-L-iduronidase (Aldurazyme). This facility is expected to supply the projected needs for Aryplase. Like Aldurazyme, arylsulfatase B is manufactured using 110 liter continuous perfusion bioreactor(s). The manufacturing process is substantially similar to that for Aldurazyme.
Some or all fill and finish operations are (before the company's manufacturing problems started in late 2009) performed under contract by Genzyme Corp. at its Allston (Boston), MA, facilities. In Nov. 2009, with particles reported in Naglazyme vials, BioMarin reportd it would be transitioning to a different fill/finish contractor.
The Master Cell Bank (MCB) and Working Cell Bank (WBC) of CSL4S-342 CHO cells (originally developed by CSL, Inc.; formerly Commonwealth Serum Labs.) were prepared at Tektagen Inc.. The bioreactor can control cell densities up to as high as 37 million cells/mL. The CHO-K1 CSL4S-342 CHO cell line expresses recombinant precursor N-acetylgalactosamine-4-sulfatase (ASB) at levels of ≥40-80 µg per 107 cells per day.
[The following information is from the European Product Assessment Report (EPAR)].
The expression plasmid pEF4S was constructed using the cDNA of rhASB and fused to the rat preproinsulin 5’ UTR to provide strong translation initiation. This plasmid contains a neomycin phosphotransferase marker gene, allowing selection of transfected cell lines using geneticin, the SV40 origin of replication and the ampicillin resistance marker gene for the initial cloning in bacteria. This pFE4S plasmid was electroporated (transfected) into CHO-K1 cells. After selection with geneticin, one stable clone was isolated and sub-cloned to obtain the CHO cell line CSL-4S-342. For the construction of the Master Cell Bank (MCB), CSL-4S-342 cells underwent a series of amplifications, and aliquots of cell suspension were processed for cryogenic storage in cryovials in the vapor phase of liquid nitrogen.
The MCB and the initial Working Cell Bank (WCB1) were prepared by Tektagen (now Charles River Labs.). WCB1 was used to produce galsulfase for all preclinical studies, Phase trials and some Phase II trials. Upon exhaustion of WCB1, WCB2 was prepared by BioReliance Corp. (now a subsidiary of Invitrogen) using different serum-free media. Fetal calf serum (FCS) and trypsin were not used in the growth and passage of the cell line CSL-4S-342, but FCS was used in the culture of the originator cell line CHO-K1. For commercial production, the MCB will only be used in case of emergency; otherwise the company uses WCB2, as it currently does. The supply of WCB2 is expected to last for ten years.
Manufacture is started by thawing a WCB2 (or MCB) vial and inoculation of its content in a cell culture flask. When the target cell density is reached, a series of amplification steps is initiated. Bioreactors are inoculated according to a parallel or sequential seeding procedure. When the target cell density in these is reached, the harvested cell culture fluid (HCCF) is collected, sampled for analysis, and stored until the first step of the purification process is initiated. Purification of rhASB from the HCCF involves: harvesting filtration to remove cells and debris and concentration using an ultrafiltration system, pH adjustment and clarification, and column chromatography. One to three initial column chromatography cycles can be combined to form a single lot, which then passes through two additional orthogonal chromatography steps, followed by low pH (acidic conditions) viral inactivation.
The product is processed by ultrafiltration/dialfiltration for concentration and buffer exchange into the final formulation buffer. Following DNA removal filtration and 0.02 µm viral removal filtration, formulation buffer is added to dilute the rhASB to a final protein concentration of 1 mg/mL and polysorbate 80 (Tween 80) is added. The bulk is transferred through a 0.2 µm filter into sterile storage containers, and can be stored up to 12 months frozen. During the purification process, pooling can occur prior to three key stages: the initial ultrafiltration, pH adjustment (in conjunction with the initial column chromatography), and the second column chromatography step. One lot is defined as the eluate from a single run of the second chromatography column, and then carried down through the process. Purification has been designed to remove process-related impurities, such as host cell proteins, retroviral particles and DNA, product-related impurities, such as aggregates and degraded products, and adventitious contaminants, such as viruses and mycoplasmas.
DNA content has been monitored during the purification of seven consecutive production lots manufactured at the Galli site using the current process. Conservative calculations in the worst case scenario meet the WHO limit of <10 ng/dose. The justification for not including host cell DNA evaluation in the release testing was considered acceptable. Residual host cell proteins (CHOP) have been determined at several points in the purification process of three consecutive production lots, using both qualitative and quantitative methods. Data were provided on the removal of glycerol (residual solvent used in the second column chromatography step) and potential column leachable materials.
With regard to product-related impurities, the three common degradation pathways for proteins such as rhASB – oxidation, aggregation, and deamidation – have been considered. Forced degradation studies have shown that rhASB is resistant to each of these pathways.
The viral safety of rhASB has been demonstrated following a series of studies to characterize the production process, raw materials, and each lot of rhASB. BioMarin is in compliance with the latest version European guidance documents concerning BSE/TSE prion contamination. No animal-sourced materials are used in the commercial-scale rhASB manufacturing process, but were used in the derivation of the host cell strain.
A number of changes were made during product development, which can be grouped in four categories: a) Cell culture: the initial process was conducted using a fed-batch process. Transition to a perfusion process was implemented to increase productivity for Phase II trials and beyond; b) Purification: modifications to the purification process, including the elimination of one chromatography step (DEAE Sepharose), optimization of the remaining chromatography steps, and increase of the diameters of all three remaining columns were made to improve the process capacity and purity of the product; c) Formulation: the addition of polysorbate 80 was implemented for Phase II trials and beyond to prevent particulate formation in the drug product that had been observed during development; and d) Facility: the process was moved from the BioMarin clinical facility to the Galli commercial facility with increased production capacity. The Galli facility process represents a modified version of the perfusion process in that longer cell culture duration and a different HCCF sublot pooling strategy are employed. The purification scheme remained unchanged except for larger column diameters for increased HCCF processing capacity. Product manufactured by the commercial process was used in pre-approval Phase II and later clinical trials.
An issue was raised during the EMEA assessment regarding the comparability between lots manufactured at the Galli site using the current commercial process and those manufactured for the preclinical and clinical studies prior to the final modifications to the manufacturing process. Satisfactory validation data were provided for three qualification lots manufactured at the Galli site with the current commercial process.
Two truncated N-terminal variants of rhASB have been identified. BioMarin considered it unlikely that the observed degree of truncation would affect safety or efficacy. C-terminal amino acid characterization also detected smaller amounts of a variant resulting from the truncation at the C-terminus of rhASB. BioMarin committed to continuing investigation of these C-terminal truncated forms and to providing a formal review once sufficient data are available. IEF studies showed that the charge heterogeneity of rhASB is conferred by oligosaccharides and that the parent protein backbone is essentially homogeneous. It appeared that rhASB has a high phosphorylated oligomannose content compared to that of sialic acid.
Potency has been determined by two methods which are now used in the release testing of the drug substance and drug product. The first is the enzymatic activity assay that utilizes 4-methylumbelliferyl sulphate (4-MUS), a small fluorigenic substrate. This assay was modified and optimized during development. The second method is the fibroblast uptake assay, a cell-based assay.
The bulk drug substance is shipped to Hollister-Stier Labs. for manufacture of the drug product. This involves final sterile filtration using 0.2 µm filters, filling into vials and preliminary labelling. Hollister-Stier was acquired by Jubilant Organosys Ltd. in April 2007.
[Most of the following is based on BioMarin’s U.S. patent application 2004131605, which is less reliable, in terms of reflecting actual manufacturing methods, than the EMEA EPAR information above].
ASB is produced using the transformed cell line cultured in JRH Excell 302 medium supplemented with L-glutamine, glucose and hypoxanthine/thymidine. The medium is further supplemented with folic acid, serine, and asparagine, and there is no G418 present in the medium. The cells are cultured in a 110 L continuous perfusion bioreactor with removal of media (harvest) starting about day nine and continuing 35-90 days. The collection rate is ~800 L per day/bioreactor.
Manufacture starts with expansion of a vial of cells from the Working Cell Bank (WCB) in a 75 mL T flask for 3 days; then a 250 mL spinner flask for 3 days; then a 1 L spinner flask for 3 days; then two 8 L spinner flasks for 1-2 days; then a 110 L bioreactor (90 L working volume) for 9 days; harvest and cell removal by filtration through a 10-membrane 1 µm and 2 µm and filter cartridge, concentration ten-fold by ultrafiltration; diafiltration using a tangential flow filtration (TFF) system; and the bulk is pumped into a 100 L bag.
The rhASB Drug Substance Purification Process involves ultrafiltration/diafiltration (UF/DF); DEAE Sepharose Fast Flow (FF) chromatography; Blue Sepharose 6 FF chromatography; copper (CU+2) Chelating Sepharose FF chromatography; Phenyl Sepharose HP chromatography; concentration by ultrafiltration and diafiltration to a final concentration of 1.5 mg/mL in formulation buffer (150 mM NaCl, 10 mM NaPO4 (pH 5.8) using a TFF system (and diluted with additional formulation buffer, if needed) to 1.0 mg/mL; final formulation with polysorbate 80 (Tween 80); sterile filtration (0.02 µm nanofiltration for virus removal) into a sterile container; storage; and filling into 5 mL Type 1 glass vials, which are manually stoppered and crimped. The four-step column chromatography including using DAEA Sepharose, Blue Sepharose, Cu+2 Chelating Sepharose and Phenyl Sepharose chromatography sequentially results in especially highly purified recombinant enzyme. Recovery yield can be >88%.
Figure 3 of U.S. 2004131605 presents the many sampling, quality control tests performed at various steps. Tests performed on the Formulated Bulk (for storage) include peptide mapping, visual assessment, endotoxin testing; SDS-PAGE; IEF; sialic acid content; oligosaccharide profiling; RP-HPLC; activity assay; in vitro uptake assay; A260; pH; and osmolality.
An important aspect of the manufacturing process is the early separation or inhibition of proteases to minimize the number of molecules of precursor ASB cleaved into the mature or processed form and/or other degraded form(s). This can be accomplished by the reduction, inhibition, or inactivation of protease activity, and the physical separation of the protease(s) from the precursor enzyme during purification. Preferably, this is performed as early as possible during the purification process. The sooner the protease activity present in the cell cultures is reduced or inhibited by pH reduction, the fewer the number of precursor molecules cleaved into the mature or processed form.
Protease activity is reduced or inhibited by adjusting the fluid to a pH value between about 4.8- 5.2 (acidification). Protease activity is found in one or more cysteine proteases, particularly in cathepsin L, that specifically cleave precursor N-acetylgalactosamine-4-sulfatase. Cathepsin L has a molecular weight of about 36 kDa in its inactive form and is converted to its active forms of 21-29 kDa in size upon exposure to pH of less than 5.0.
The purity of precursor N-acetylgalactosamine-4-sulfatase is measured using reverse-phase high performance liquid chromatography (RP-HPLC), which separates proteins based on differences in hydrophobicity. This assay uses a C4 column (Phenomenex Jupiter) as the stationary phase and a gradient of water:acetonitrile as the mobile phase.
Preliminary drug product specifications presented in the patent application include [presented in format Test - Procedure - Specification] – activity - fluorescence assay - 20,000-120,000 M units; adventitious viruses - in vitro assay (tested on harvested supernatant from bioreactor) - pass; appearance - visual - clear, colorless to pale yellow solution; bacterial endotoxin - LAL - ≤2 eu/ml; chloride - atomic absorption - report value; ASB fibroblast uptake - tbd - ≤40 nMol; mycoplasma assay (tested on harvested supernatant from bioreactor) - points to consider - pass; particulates - USP - ≤600/vial at 25 µM and ≤6000/vial at 10µM; pH - USP - 5.5 - 6.8; phosphate - atomic absorption - report value; protein concentration - UV 280 - 0.8-1.2 mg/mL; purity - SDS page - 1 major band between 65-70 kDa; RP - HPLC - >95%; residual blue dye - tbd - report value; residual copper - tbd - report value; sodium - atomic absorption - report value; specific activity - calculation - 40,000-80,000 mUnits/mg; sterility - 21 CFR 610 - pass.
FDA class: Biologic BLA
Approvals: Date = 20050531; original BLA with orphan status; Indication = mucopolysaccharidosis VI
Indications: [Full text of the "Indications AND USAGE” section of product insert/labeling]:
Naglazyme is indicated for patients with Mucopolysaccharidosis VI (MPS VI). Naglazyme has been shown to improve walking and stair-climbing capacity.
Status: On Nov. 29, 2004, BioMarin submitted a digital-format BLA for Naglazyme as an enzyme replacement therapy for the treatment of mucopolysaccharidosis VI (MPS VI). This was accepted on Feb. 1, 2005 and granted priority review (6 month PDUFA/target date of May 31, 2005). Naglazyme had previously received fast track status and orphan drug designation for this indication. Approval was granted on June 1, 2005 (approval time = ~6 months) with orphan status. BioMarin launched Naglazyme in the U.S. on June 21, 2005.
On Dec. 6, 2004, BioMarin submitted a Marketing Authorization Application (MAA) for European Union (EU) approval with orphan medicinal product designation. Approval was granted on Jan. 24, 2006 for long-term enzyme replacement therapy in patients with a confirmed diagnosis of MPS VI with orphan status. Naglazyme become the first specific treatment approved in the European Union for patients with MPS VI.
BioMarin is conducting post-approval studies, including use of Naglazyme in children under the age of 5; monitoring to determine if patients develop antibodies which could affect the response to treatment; and to determine the optimal dose for regular long-term use.
In Aug. 2007, AnGes MG, Inc. filed a BLA in Japan for MPS VI. Approval was granted on March 31, 2008, bringing the first treatment option to MPS VI patients in Japan.
Tech. transfer: Patent applications covering aspects of Aryplase assigned to BioMarin include US2004131605, “Precursor of N-acetylgalactosamine-4-sufatase, methods of treatment using said enzyme and methods for producing and purifying said enzyme,” filed Nov. 7, 2003. U.S. priority extends to May 1, 2000. This concerns highly purified recombinant human precursor N-acetylgalactosamine-4-sulfatase (and active mutants, fragments and analogs), methods for producing and purifying the recombinant precursor enzyme to a highly purified form, and formulations for treatment of MPS VI. Claims include precursor enzyme with a purity ≥99.9% based on total protein by reverse-phase HPLC; use of transfected CHO CSL4S-342 cells in supplemented JRH Excell 302 medium; purification of precursor N-acetylgalactosamine-4-sulfatase from culture fluid by reducing the proteolytic activity of a protease to cleave the molecule; purification using a Cibracon blue dye interaction chromatography resin (Blue Sepharose 6 Fast Flow column), a copper chelation chromatography resin (Chelating Sepharose Fast Flow column) and then a phenyl hydrophobic interaction chromatography resin (Phenyl Sepharose 6 Fast Flow High Sub column), and recovery of precursor enzyme.
Presuming BioMarin’s patent applications in the U.S. and other countries are granted, the company will have relatively long patent-based exclusivity, compared to many products which receive patents years prior to commercialization.
Trials: BioMarin performed three clinical trials with Aryplase and one disease survey study.
In June 2004, BioMarin reported positive results from its 24-week double-blind Phase III trial of Aryplase. The trial enrolled 39 patients ages 5-29 years at six sites around the world. Patients were randomized to receive a weekly intravenous infusion of 1.0 mg/kg of either galsulfase or placebo solution for 24 consecutive weeks. A statistically significant improvement in endurance (p = 0.025) was observed in patients receiving galsulfase compared to patients receiving placebo as measured by the distance walked in 12 minutes. This was the primary endpoint in the trial. A statistically significant reduction in GAGs excreted in the urine (p < 0.001) was reported in patients receiving galsulfase compared to patients receiving placebo, a surrogate marker of enzymatic bioactivity. GAGs reduction was one of two secondary endpoints. The 3-minute stair climb, another measure of endurance and the other secondary endpoint, demonstrated a positive trend (p = 0.053) in patients receiving galsulfase compared to patients receiving placebo. Aryplase was generally well-tolerated. Adverse events during infusions were more common in patients receiving Aryplase, but were generally mild to moderate. The frequency of serious adverse events was more common in the placebo group. Nearly all patients developed galsulfase antibodies as a result of treatment, but the level of the immune response did not correlate with adverse events or impact the improvements in endurance. Following the trial, all patients began receiving weekly infusions of galsulfase in an open-label extension study with continued assessment of primary and secondary efficacy variables as well as assessments related to safety and tolerability.
In March 2005, BioMarin reported results from its extension study. This included all 38 patients who completed the initial 24-week trial. Patients who received galsulfase for an additional 24 weeks, for a total of 48 weeks, continued to experience improved endurance, and patients who initially received placebo, and then received galsulfase for 24 weeks also experienced improved endurance.
As for all lysosomal genetic disorders, including MPS VI, it is of primary importance, especially in severe forms, to initiate treatment as early as possible, before appearance of non-reversible clinical manifestations of the disease. However, a key unresolved issue with Naglazyme is treatment of patients aged <5 years suffering from a severe form of the disease (a population not included in the pivotal Phase 3 study).
Medical: The recommended dosage regimen for galsulfase is 1 mg/kg body weight administered once every week as an intravenous infusion over 4 hours. The initial infusion rate is adjusted so that approximately 2.5% of the total solution is infused during the first hour, with infusion of the remaining volume (~97.5%) over the next 3 hours.
Disease: MPS VI (mucopolysaccharidosis VI; Maroteaux-Lamy syndrome; polydystrophic dwarfism) is an inherited debilitating, life-threatening disease for which no therapies are currently available other than Naglazyme. As discussed in the Biological section above, MPS VI is caused by genetic deficiency of N- acetylgalactosamine 4-sulfatase (arylsulfatase B), a lysosomal enzyme normally required for the breakdown of certain complex carbohydrates known as glycosaminoglycans (GAGs). This enzyme deficiency leads to the accumulation of GAGs in the lysosomes of cells, giving rise to progressive cellular, tissue and organ system dysfunction. In the absence of the enzyme, the stepwise degradation of dermatan sulfate (and other GAGs) is blocked and dermatan sulfate accumulates intracellularly in the lysosome in a wide range of tissues. This causes a progressive disorder with multiple organ and tissue involvement in which infants appear normal at birth, but usually die before puberty. Unlike MPS I, MPS VI is not typically associated with progressive impairment of mental status, although physical limitations may impact learning and development.
The diagnosis of MPS VI is usually made at 6-24 months of age when children show progressive deceleration of growth, enlarged liver and spleen, skeletal deformities, coarse facial features, upper airway obstruction, and joint deformities. Progressive clouding of the cornea, communicating hydrocephalus, and/or heart disease may develop in MPS VI children. Death usually results from respiratory infection or cardiac disease. Debilitating symptoms can include impaired cardiac and pulmonary function, delayed physical development, skeletal and joint deformities, impaired vision and hearing, sleep apnea, and reduced endurance. The majority of people with MPS VI die from disease-related complications between childhood and early adulthood. Patients with a less severe form of the disease may survive for decades.
A consensual estimate is that there are between 50 and 300 patients in the U.S. diagnosed with all forms of MPS VI. An estimated 1,100 people worldwide have the condition. BioMarin has estimated that there are only slightly more than 200 U.S. patients with MPS VI who could potentially benefit from Naglazyme treatment.
Market: Worldwide revenue was $192.7 million in 2010; $168.7 million in 2009; $132.7 million in 2008; and $86 million in 2007. Net sales of Naglazyme were $46.5 million in 2006 (first full calendar year on the market). In early 2011, BioMarin reported that 37% of sales of Naglazyme derive from outside of the U.S. and Europe.
The Average Wholesale Price (AWP) reported in the 2007 edition of the Red Book is $1,812/vial with a Direct Price (DP; bulk discount) of $1,450.
Naglazyme treatment costs an average of over $20,000/year, similar to other ultra-orphan enzyme replacement therapies.
BioMarin markets Naglazyme worldwide using its own sales force, including operating its own marketing subsidiary in Europe. BioMarin has launched BioMarin Physician and Patient Support (BPPS), a free confidential service to assist patients and caregivers in receiving insurance reimbursement for treatment with Naglazyme.
With ~200 MPS VI patients in the U.S., Aryplase is very much an orphan product. It can be presumed that essentially all MPS VI patients receive Naglazyme. Marketing is targeted to those specialists who treat MPS VI patients. The market outside the U.S. can be presumed to primarily be in Europe, Japan and other developed countries affluent enough to afford Naglazyme treatment of MPS VI patients. This market foreign may roughly be presumed to be comparable in size to the U.S. market (~200 patients).
No other MPS VI therapeutics are known to be in clinical trials, so Naglazyme will have no competition for quite some time.
Companies involvement:
Full monograph
105 Arylsulfatase B, rDNA
Nomenclature:
Arylsulfatase B, rDNA [BIO]
Aryplase [TR former]
Naglazyme [TR]
galsulfase [USAN INN]
N-acetylgalactosamine 4-sulfatase (human CSL4S-342 cell) [CAS]
sulfatase, acetylgalactosamine 4- (human CSL4S-342 cell) [CAS]
552858-79-4 [CAS RN]
4-sulfatase [SY]
arylsulfatase B [SY]
ASB [SY]
BM-102 [SY]
chondroitin 4-sulfatase [SY]
chondroitinase [SY]
chondroitinsulfatase [SY]
G4S [SY]
MPS6 [SY]
N-acetylgalactosamine 4-sulfatase [SY]
nitrocatechol sulfatase [SY]
rhASB [SY]
EC 3.1.6.12 [EC]
molecular weight (kDa) = 56
FDA Class: NDA Drug
Year of approval (FDA) = 2005
Date of 1st FDA approval = 20050531
(in format YYYYMMDD)
Biosimilars/biobetters-related U.S. Patents: | 2022-2028, according to BioMarin; 2022 based on 6,866,844) |
U.S. Patent Expiration Year: | 2022 |
U.S. Biosimilars Data Exclusivity Expiration: | 2017 |
U.S. Biosimilars Orphan Exclusivity Expiration: | 2012 |
U.S. Biosimilars Launchability Year: | 2022 |
U.S. Biobetters Launchability Year: | 2022 |
Biosimilars/biobetters-related EU Patents: | 2022-2028, according to BioMarin |
EU Patent Expiration Year: | 2022 |
EU Biosimilars Data Exclusivity Expiration: | 2014 |
EU Biosimilars Orphan Exclusivity Expiration: | 2014 |
EU Biosimilars Launchability Year: | 2022 |
EU Biobetters Launchability Year: | 2022 |
Index Terms:
BHK-21 (C-13)
enzyme replacement therapy (ERT)
exempt from CBER lot release requirements
hamster proteins
murine (mouse) hybridoma cells
recombinant DNA
rodent cells <!-- rodentcells -->
ampicillin
asparaginase, L-
bioreactors, 10,000 Liter
bioreactors, 10,000 Liter
castor oil
chicken source materials
chlortetracycline
chymopapain
Cryoprecipitated AHF (antihemophilic factor)
Eu human monoclonal antibody
Flury LEP (Low Egg Passage) C25, rabies virus
genetic diseases
glutamine synthetase (GS) expression system
glycosaminoglycans
JM101, Escherichia coli (E. coli)
JM101, Escherichia coli (E. coli)
mannose
media, serum-based
neomycin
percutaneous transluminal coronary angioplasty (PTCA)
petrolatum gauze
plasmid pDW27
PrefGel
protamine
radioimmune conjugates<!-- radioconjugates -->
Sepharose, Erythrina trypsin inhibitor (ETI)-
Serum (Human), immunoglobulin-depleted
suspension cell culture
4-acetylphenylacetic acid
blood transfusion
chondrocytes, human
conjugates
DEAE Sepharose
deoxyribonuclease (DNase)
fibrinolysin
glucosamine
glycerinated-water-potato medium
Namalva cells
phenol red
phenthioate ligand
polysorbate 60 (Tween 60)
Sepharose
sodium carboxymethylcellulose
sodium phosphate
sodium phosphate, dibasic
Tac subunit of IL-2 receptor
vesicular stomatitis virus (VSV)
Viral Antigen Free mice
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
octoxynol (Triton X-100)
PrefGel
EU200 Currently Approved in EU
UM001 Marketed Product in US
US200 Currently Approved in US
EM001 Marketed Product in EU
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