Status: approved; marketed, negligible sales

Organizations involved:

Genzyme General – Manuf.; R&D; Tech.; USA mark.

Genzyme Corp. – Intl. mark.; Parent

Sanofi Pasteur S.A. – Manuf. other

Cross ref: See the entry in the Recombinant DNA section for Imiglucerase (Cerezyme) (#167), a recombinant form of this enzyme, also from Genzyme which has essentially replaced this product, for further background and other information.

Description: Alglucerase or Ceredase is an aqueous formulation of beta-glucocerebrosidase (beta-D-glucosyl-N-acyl-sphingo-sine glucohydrolase; EC 3.2.1.45) glycoprotein enzyme purified from human placenta tissues, with enzymatic modification of the molecule’s glycosylation (carbohydrate side chains). Modification involves replacing the carbohydrate residues at the non-reducing ends of the oligosaccharide chains of the glycoprotein so that they are predominantly terminated with mannose residues, which are specifically recognized by carbohydrate receptors on monocyte/macrophage lineage (e.g., Gaucher) cells. manufacture of alglucerase includes extraction and purification with solvents and detergents, resulting in substantial viral inactivation (8-30 log10 for viruses tested).

This product has substantially been replaced by a side chain-modified recombinant form of beta-glucocere-brosidase, imiglucerase (Ceredase), also manufactured and marketed by Genzyme General.

The source enzyme is purified from a large pool of human placental tissue collected from selected donors (women having just delivered babies). Placental glucocerebrosidase has glycosylation patterns different than that of the enzyme found in human liver, spleen, brain, or macrophages. The glycosylation patterns of the native human beta-glucocerebrosidase glyco-protein are modified by subjecting it sequentially to neuraminidase enzyme to remove terminal sialic acid residues, beta-galactosidase enzyme to remove galactose residues, and beta-N-acetylglucosamidase enzyme to remove N-acetylglu-cos-amine residues. The resulting beta-glucocerebrosidase (alglucerase) has oligosaccharide chains predominantly terminating in mannose residues. This provides a significantly longer in vivo half-life and higher affinity for cellular mannose receptors, e.g., on macrophages including Gaucher cells.

Alglucerase is a monomeric glycoprotein of 497 amino acids (MW = 59,300 as determined by SDS-PAGE) with carbohydrates making up approximately 6% of the molecule. The unmodified human enzyme (beta-glucocerebrosidase) also contains 497 amino acids (MW = 67,000) and has approximately 12% carbohydrate. The carbohydrates on the unmodified enzyme consist of N-linked carbohydrate chains of the complex and high mannose type.

Alglucerase, like native beta-gluco-cere-brosidase and recombinant imiglucerase (Cerezyme), catalyzes the hydrolysis of the glycolipid, glucocerebroside, within the lysosomes of the reticuloendothelial system, particularly macrophages, yielding glucose and ceramide. Alglucer-ase was the first mannose-terminated beta-glucocerebrosidase product (with improved potency and efficacy) for enzyme replacement therapy of Gaucher disease.

Ceredase is packaged in bottles containing 400 and 50 units per bottle, with a fill volume of 5 mL per bottle. The 400 unit bottle contains alglucerase at a concentration of 80 units/mL; and the 50 unit bottle has a concentration of 10 units/mL. One beta-glucocerebrosidase enzyme unit (U) is defined as the amount of enzyme that catalyzes the hydrolysis of one micromole of the synthetic substrate para-nitrophenyl-beta-D-glucopyranoside (pNP-Glc) per minute at 37˚C.

The originally approved product was formulated with Albumin (Human), but this is not used in the currently.

Nomenclature: Glucocerebrosidase [BIO]; Ceredase [TR]; alglucerase [FDA USAN INN BAN]; glucosyl-ceramidase (human placenta isoenzyme protein moiety reduced) [CAS]; ceram-idase, glucosyl- [CAS]; 143003-46-7 [CAS RN]; 37228-64-1 [CAS RN]; beta-D-glucosyl-N-acylsphingosine glucohydrolase [SY]; beta-glucocerebrosidase [SY]; glucocerebrosidase, beta- [SY]; mannose-terminated human placental glucocerebrosidase [SY]; macrophage-targeted beta-glucocerebrosidase [SY]

Companies.: Ceredase was developed and is manufactured by Genzyme General, a subsidiary of Genzyme Corp. Ceredase is marketed by Genzyme General in the U.S. and internationally by Genzyme affiliates. Development of Ceredase was at least partially funded through Ceredase Partnership (a limited partnership or other special purpose funding entity).

Source human placenta tissue is processed and provided under contract by Imedex, a subsidiary (at time of approval) of Institut Merieux, which became, Pasteur Merieux Connaught S.A., now Aventis Pasteur S.A., now Sanofi Pasteur S.A.

Manufacture: Ceredase manufacture includes stringent product testing performed both in process and on the final product. Sophisticated manufacturing, quality control, monitoring and controls have been developed. This includes use of a computerized statistical database analysis/process control system capable of tracking, detecting and adjusting for minute variations in the chemical composition of the enzyme in process and in the manufacturing equipment. The company reports, “In sensitivity and sophistication, this is believed to be unique in the pharmaceutical industry.”

Ceredase is prepared from selected placental tissue from healthy pregnant female donors with normal, full-term deliveries, primarily from North America and Europe. Placentae associated with abortions, excessive meconium, or stillbirths are not accepted. Placentae selection, preliminary processing and enzyme extraction are performed under contract by Imedex, a subsidiary (at time of approval) of Institut Merieux, now Aventis Pasteur S.A., which also processes placentae for preparation of Albumin (Human) and other products. Placentae are collected at the time of delivery, stored frozen at -20˚C, and transported to Imedex in France frozen in refrigerated containers at temperatures below -15˚C. Placentae received by Imedex are inspected and those with signs of defrosting are rejected. Accepted placentae lots are stored in a cold room at below -15˚C. The placentae are cleaned, fluid is removed by pressure filtration, and placentae are dried. All further processing of the enzyme extract, including carbohydrate modification and purification, are performed by Genzyme. Preparation was described in Murray, et al., Biochim. Biophys. Acta 673, 425, 1981; and Meth. Enzymol. 149, 25, 1987.

beta-Glucocerebrosidase is extracted and then purified from dried placentae using mixtures of solvents and detergents at Genzyme facilities. Similar to solvent detergent viral inactivation used for many human blood-derived products (see the Plasma SD entry #799 for further information), exposure to solvents and detergents substantially disrupts and inactivates lipid enveloped viruses (e.g., HIV), and the solvent extraction also removes many viruses. Ceredase is heat labile and cannot withstand heat inactivation (pasteurization). The extracted, virus-inactivated, bulk enzyme is further extracted and purified by precipitation, ultrafiltration, and chromatography.

The oligosaccharide side chains of the enzyme are modified by proprietary sequential treatment with neuraminidase, beta-galactosidase, and beta-N-acetylglucosamidase enzymes to remove terminal sialic acid, galactose and N-acetylglucos-amine residues, respectively. [The recombinant enzyme, imi-glu-cerase, is similarly processed]. Excipients are added and the product is formulated, sterile filtered, and bottled.

Testing, both in-process and final, includes tests for HIV-1, HIV-1 and HIV-2 antibodies, hepatitis B virus surface antigen (HBsAg). Antibody and antigen testing are performed using ELISA immunoassays. Four extraction/purification steps in the manufacturing process have been validated for inactivation/removal of a wide spectrum of human viruses, including retroviruses, DNA, RNA, enveloped and non-enveloped viruses. The panel of viruses consisted of HIV-1, hepatitis B virus (HBV), hepatitis C virus (HCV; then called non-A, non-B hepatitis virus), poliovirus, herpes simplex virus (HSV), and rabies virus. The four solvent/detergent extraction/purification steps cumulatively removed from 8-30 log of virus from spiked samples.

Genzyme discovered in Sept. 1993 that small amounts of human chorionic gonadotropin (hCG; about 3 units of hCG activity per unit Ceredase), a hormone present in placen-tae, were present in Ceredase. The hCG was partially deglyco-sylated by the Ceredase manufacturing process, with deglyco-sylated hCG having a clearance rate in vivo about 40 times that of native human hCG. Genzyme made improvements to the manufacturing process resulting in a 15-fold decrease in hCG. The level of hCG in the product is now less than 1 µg hCG per mg alglucerase protein, as determined by ELISA assay.

FDA class: Drug NDA

Approvals: Date = 19910405, first approval, NDA; orphan designation (expired 4/1998)

Date = 19970108; NDA supplement; Indication = modifications of product manufacture and labeling to include reduction of residual levels of human chorionic gonadotropin (hCG)

Indications: [full text of the "INDICATIONS AND USAGE” section from product insert/labeling]:

Ceredase (alglucerase injection) is indicated for use as long-term enzyme replacement therapy for patients with a confirmed diagnosis of Type 1 Gaucher disease who exhibit signs and symptoms that are severe enough to result in one or more of the following conditions: moderate-to-severe anemia; thrombocytopenia with bleeding tendency; bone disease; [and] significant hepatomegaly or splenomegaly.

Status: No centralized European Union approval ever granted.

Tech. transfer: Patents expired. No entries for Ceredase in the Orange Book.

Medical: See the imiglucerase (Cerezyme) entry #167 for information about Gaucher disease.

Market: The 2007 Average Wholesale Price (AWP) is $1,580.00/vial, with a Direct Price (Manufacturer’s discount price) of $1,480.00 (Red Book, 2007). These prices are unchanged from 2004.

Cerezyme (recombinant enzyme), also from Genzyme General, has largely replaced Ceredase. Ceredase is still approved and manufactured, but the company does not actively promote the product. Ceredase has few, if any, additional benefits compared to the recombinant product; and, theoretically, entails significantly higher risks, e.g., viral contamination. It may still be needed and used by those with allergy or intolerance to components in the recombinant product.