COMPANY PROFILE
Celsus, Inc., a closely-held enterprise, was founded in 1987 to manufacture bulk heparin derivatives and
to pursue the development of other complex macromolecules belonging to the family of the sulfated
glycosaminoglycans. The Company's tradename evokes the name of Aulus Cornelius Celsus, a first century
Roman medical encyclopedist.
Celsus Laboratories, Inc., a wholly-owned subsidiary company, is a USDA-approved, FDA-registered
drug establishment with the capacity to manufacture and test heparin. Heparin chemistry has multiple
applications including the use in the collection and analysis of blood; in non-thrombogenic coatings for
coronary stents and other blood-interacting medical devices; in the prevention and treatment of thromboembolic disorders; in end-stage renal dialysis and in cardiac surgery. Heparin also serves as the affinity ligand in the purification of many bio-engineered heparin-binding proteins, and as the raw material for low molecular weight heparins and heparin lyase, used clinically.
Celsus Biopharmaceuticals, Inc., is an affiliated company dedicated solely to the synthesis and preclinical
development of Intimatan™, an improved anticoagulant for use in cardiac surgery. Intimatan™ is a
patented polysaccharide comprising repeating L-iduronic acid 4,6-O-disulfated N-acetyl-D-galactosamine
disaccharide units. Preclinical pharmacology studies, thus far, have shown Intimatan to be an effective
inhibitor of surface-bound thrombin, complement activation and neointimal hyperplasia in models of
restenosis injury. In the treatment of acute coronary thrombosis, Intimatan reduces the dose of platelet
GPIIb/IIIa inhibitors required to block coronary thrombosis in vivo and prevents recurrent coronary artery
thrombosis in the canine following adjuvant thrombolysis with tissue plasminogen activator. It completely
prevents primary arterial and venous thromboses in the canine model of electrolytic injury. In the pig
model of cardiac surgery, Intimatan maintained extracorporeal patency at one tenth the anticoagulant
dose; generated a 4-fold lower activated clotting time (ACT); reduced chest wall bleeding more than 2-
fold; and did not induce thrombin rebound or require neutralization post-procedure. Intimatan also
prevents inflammation damage to the isolated perfused rabbit heart and may thus protect against the
ravages of ischemic-reperfusion injury in myocardial infarction and stroke. Furthermore, Intimatan blocks
the activation of platelets of patients with heparin-induced thrombocytopenia (HIT) caused by heparin
allergy. As a HIT antagonist, Intimatan may benefit high-risk cardiac patients who experience thrombosis
due to heparin exposure.
Administrative Offices & Drug Establishment
12150 Best Place, Cincinnati, Ohio 45241-1569 USA
For manufacture and sale of its bulk drug substances, Celsus is subject to the requirements of the
Federal Food, Drug and Cosmetic Act and the regulations administered by the FDA. The laws and
regulations require periodic inspections and compliance with current Good Manufacturing Practices.
cGMP is an allowable alternative to ISO-certification under the EU Medical Device Directive.
The Company is FDA-registered (1527761) to manufacture bulk drug substances in an establishment
which has been approved by the Animal & Plant Health Inspection Service of the U.S. Department of
Agriculture to handle restricted animal by-products. Confidential drug master files have been submitted to
regulatory authorities in Australia, Canada, the European Union and the United States and are available
for reference, as appropriate.
The porcine raw material from which Celsus’ products are derived is sourced in accordance with EU
regulatory guidelines prEN 12442-2 from healthy animals, raised and slaughtered for food by man in
compliant member nations of the Office International des Epizooties that are free of bovine transmissible
spongiform encephalopathy (TSE). In addition, the raw material is first screened for the absence of prion
antigen reactivity by immunoassay sensitive to ~ 1ppm detection, and is then purified using a process
validated for the removal of TSE infectious agents and viral infectivity due to retroviruses, enterohepatic
viruses and porcine parvovirus. These studies are summarized below.
To validate the process, model viruses were chosen in compliance with EU regulatory guidelines prEN
12442-3 and the ICH Harmonized Tripartite Guidelines for Viral Safety Evaluation (see Table 1 for study
details). Model virus selection was based on the following criteria from which the robustness of the
manufacturing process on viral elimination was gauged: (i) potential pathogenic relevancy to porcinederived
heparin sodium; (ii) genomic diversity of virus classifications; (iii) physical-chemical diversity of
virus structural and functional properties and; (iv) relative resistance to physical-chemical inactivation
procedures from very low to very high resistance. Two steps of the heparin manufacturing process were
investigated. The first step was an extended exposure to oxidation. The second one involved a limited
exposure to oxidation. The virus spiking protocols utilized intermediate stage heparins derived from
porcine raw material that tested negative for prion antigens. To evaluate these processing steps for virus
reduction, intermediate stage heparins were pulled from production, spiked with the model viruses to a
high titer, subjected to the respective purification steps of the batch record and then evaluated for virus
survival.
Extended Exposure Protocol
For the HIV studies 5 x E6/ml of strain RF was used to test the resistance of HIV to an extended
exposure to heat, alkaline pH and inorganic peroxide. A ≥ 6.2 log reduction was obtained (see Table 2).
For the polio studies, virus titers of poliovirus type 1 (VLS082802) ranged from 0.1–1.2 x E9/ml (average
~ 5 x E8/ml). No virus could be detected after treatment by immunofluorescence method. Relative to the
positive control, a ³ 8log reduction was computed as a conservative determination after adjustment for
toxicity and interference considerations.
Parvovirus BL-006 (NADC-2) involved ~ 1 x E6.8/ml with a ≥ 4.1 log reduction determined relative to nontreated spiked samples after adjustment for cell toxicity and interference considerations. Again, no
infectious virus could be detected using a highly sensitive immunofluorescence plaque assay method.
Limited Exposure Protocol
Three different viral strains were used (RF, IIIB and the human clinical isolate M301) to test the resistance of HIV to a limited exposure to chemical inactivation. Depending on the virus used, titers ranged from about 1.5-5 x E6 infectious units per ml. In these studies, a ≥ 5 log reduction was calculated on the basis of no recovery of infectious particles relative to the positive control taking into account toxicity and interference effects.
For the polio studies, virus titers of poliovirus type 1 (Mahoney) ranged from 0.1 – 1.2 x E8/ml (average ~
5 x E7/ml). No virus could be detected after treatment. Relative to the positive control, a ≥ 5 log reduction was computed as a conservative determination.
Parvovirus BL-006 (NADC-2) involved ~ 1 x E6/ml input of virus. A ≥ 4.3 log reduction was determined
relative to positive control after taking into account cytotoxicity effects. Again, no infectious virus could be detected using a highly sensitive immunofluorescence plaque assay method.
Although not considered a virus under the ICH Guidelines, the strain 263K Scrapie was used as a model
of the Transmissable Spongiform Encephalopathy (TSE) agent to evaluate the ability of the limited
exposure protocol to eliminate this most intractable agent. The process was evaluated by a quantal-based
assay involving the intracerebral innoculation of Syrian hamsters with heparin samples spiked with
TSE either treated or not treated by limited exposure to organic peroxidation. Samples were spiked at ~ 1
x E6/ml and evaluated by serial dilutions down to 1 infectious unit/hamster. The results of the 1 year
study showed that hamsters that received the Celsus heparin that had been treated for TSE removal by
the manufacturing process did not develop the disease. All hamster groups receiving heparin spiked with
TSE that was not treated by the Celsus process developed disease. The brains of non-infected and
infected animals from the two groups were analyzed by monoclonal antibody (Western Blot) for the
presence of the prion antigen PrPsc (data not shown). None of the animals receiving heparin that was
spiked and chemically inactivated by the limited exposure protocol had evidence of TSE 263K prion
antigen, i.e., they were free of infectious prions in brain matter.
The following tables summarize the findings of the above studies. Both the extended and limited
exposure protocols resulted in the complete eradication of all detectable traces of the infective agents
studied.
Celsus uses compendial and other validated laboratory test procedures for the certification of the
strength, quality and purity of its products. With the exception of the determination of nuclear magnetic
resonance spectra, all laboratory tests including the following are performed in-house:
Anticoagulant Activity - The amount that will cause 1 ml of plasma to half-clot when kept for 1 h at 20º C. is determined using the heparin assay, as described in the U.S. Pharmacopeia (USP).
Bacterial Endotoxins - The Limulus Amebocyte Lysate (LAL) assay is universally accepted as a reference test for the presence of bacterial endotoxins and is used to certify product as conforming to USP <85>. In addition, Celsus employs a sensitive and accurate kineticturbidimetric KTA2 assay to test Purified Water used in manufacture of its products and to compare with the LAL gel-clot method.
Prion antigens - An immunoassay for prion antigens has been in place since 1999 and utilizes a monoclonal antibody specific to an epitope conserved among bovine, ovine and porcine species. It crossreacts with both PrPsc and PrPc. Antibody specificity was demonstrated by Celsus using recombinant bovine PrP. Spiking studies with both soluble recombinant bovine PrP and particulate tissue-associated PrP from porcine brain authenticated prion detection in crude heparin to ~ 1 ppm limit.
Celsus has adopted the policy that achievement of high quality manufacture and controls comes from a
rigorous commitment to operations in conformity with current Good Manufacturing Practices (cGMP). No
product is dispatched by Celsus unless certified by Quality Assurance, an independent function duly
authorized to audit and enforce the Company’s conformity with cGMP.
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Anti-Factor Xa and Anti-Factor IIa activities - These tests are performed in vitro using amidolytic assays in a purified system on an ACL 300 Plus to measure the ability of products to accelerate the inhibition of thrombin and factor Xa by antithrombin III and heparin cofactor II.
Molecular Weight - Molecular weight distribution is determined using high-performance size exclusion liquid chromatography and oligosaccharides as calibration standards.