Abciximab - ReoPro; Platelet monoclonal antibody Fab fragment, recombinant
Status: approved; marketed
Organizations involved:
Centocor B.V. – R&D
Centocor, Inc. – Tech.; USA mark.
Johnson & Johnson Co. – Parent
Lilly, Eli & Co. – USA mark.; Intl. mark.
Novartis AG - Manuf.
State Univ. of New York (SUNY) – R&D; Tech.
Lonza Biologics plc – Tech.
Celltech Biologics plc – Former
Genentech, Inc. – Tech
City of Hope National Medical Center – Tech.; Patent dispute.
Royalty Pharma, AG – Tech.
University of Glasgow – Tech.
Fujisawa Pharmaceuticals, Inc., Ltd. - Japan mark.
Cross ref.: See Monoclonal Antibodies (entry #300).
Description: ReoPro is an aqueous formulation of abciximab, an enzymatically-derived Fab fragment of a recombinant chimeric human-murine monoclonal antibody (c7E3) glycoprotein produced by a transformed murine myeloma Sp2/0 cell line. The Fab fragment is purified from cell culture supernatant by a series of steps including specific viral inactivation and removal procedures, digestion of the monoclonal antibody with papain, and column chromatography. Abciximab binds to (is an agonist for) the glycoprotein IIb/IIIa (GPIIb/IIIa) fibrinogen receptor on human platelets, and inhibits platelet aggregation and resulting thrombus or blood clot formation.
The source chimeric immunoglobulin or monoclonal antibody, c7E3, is a dimer comprised of chimeric heavy and light immunoglobulin chains with a molecular weight of 47,615 Dalton (47.65 kDa). The chimeric heavy chain is comprised of an antigen-binding region derived from the heavy chain of murine monoclonal antibody 7E3 (hybridoma cell line ATCC HB8832), specific for the GPIIb/IIIa receptor, linked to a human heavy chain constant region. The chimeric light chain comprises the same antigen-binding region derived from the light chain of murine monoclonal antibody 7E3, specific for the GPIIb/IIIa receptor, linked to a human-derived light chain constant region. This recombinant monoclonal antibody is expressed in transformed Sp2/0 myeloma cells, recovered, and the Fab fragment (c7E3 Fab; abciximab) is derived by digestion with the proteolytic enzyme papain (e.g., derived from papaya; see entry #622).
ReoPro is a clear, colorless, sterile, non-pyrogenic solution for intravenous use. ReoPro is supplied in single-use 5 mL vials containing 10 mg of abciximab. Each vial of ReoPro contains 2 mg/mL of Abciximab in a buffered solution (pH 7.2) of 0.01 M sodium phosphate, 0.15 M sodium chloride and 0.001% polysorbate 80 in Water for Injection. The product contains no preservatives. Vials should be stored at 2-8˚C (36-46˚F; refrigerated).
Nomenclature: Platelet Mab Fab, rDNA [BIO]; ReoPro [TR reg. by Lilly]; CenteoRx [TR; former in U.S.]; abciximab [FDA USAN INN BAN]; immunoglobulin G, (human-mouse monoclonal c7E3 clone p7E3VHhCgamma4) Fab fragment anti-human glycoprotein IIb/IIIa receptor), disulfide with human-mouse monoclonal c7E3 clone p7E3VkhCklight chain]; [CAS]; 143653-53-6 [CAS RN]; Fab fragment of the chimeric human-murine monoclonal antibody 7E3 [SY]; anti-GPIIb/IIIa monoclonal antibody [SY]; GPIIb/IIIa monoclonal antibody [SY]; human-murine monoclonal antibody 7E3 [SY]; c7E3 Fab fragment [SY]
Note, terms indicative of the parent chimeric (monoclonal) antibody, and even terms simply referring to platelet (monoclonal) or GPIIb/IIIa antibody, are often used to refer to abciximab, the chimeric Fab fragment. It is also common for the term Regranex to be used ambiguously or imprecisely to refer to the active ingredient, and for platelet or GPIIb/IIIa (monoclonal) antibody terms to be used to refer to the formulated product.
Biological.: Thrombosis (blood clot formation) begins with the adhesion of platelets at sites of blood vessel wall injury. The adhesion of platelets is mediated by platelet surface receptors which bind to extracellular matrix proteins in the exposed subendothelium, such as von Willebrand’s factor, collagen, fibronectin, vitronectin, and laminin. The binding of adhesive molecules, such as fibrinogen or von Willebrand’s factor, to GPIIb/IIIa receptor causing aggregation of platelets is a common step in thrombus formation, making the GPIIb/IIIa receptor an attractive target for therapeutic agents. When blood platelets encounter injured tissue site, such as produced by percutaneous transluminal coronary angioplasty (PCTA), they adhere to it, forming a thin protective monolayer. Subsequently, platelet activation occurs in response to agonists such as epinephrine, ADP, collagen, and thrombin. Activation leads to the exposure of the glycoprotein IIb/IIIa receptor (GPIIb/IIIa) on the platelet surface. GPIIb/IIIa on activated platelets is available to bind to fibrinogen, which mediates platelet aggregation and clot formation.
The source murine 7E3 monoclonal antibody, the derived chimeric monoclonal antibody (c7E3), and the c7E3 Fab fragment (abciximab) have binding specificity for the complexed form of the GPIIb/IIIa receptor on the surface of platelet cells. Abciximab binds to (is an agonist for) the glycoprotein IIb/IIIa (GPIIb/IIIa) receptor on human platelets, resulting in inhibition of platelet aggregation and blood clotting (thrombosis). GPIIb/IIIa is a platelet cell surface molecule that normally binds to fibrinogen and von Willebrand’s factor (vWF) and mediates platelet aggregation, a critical step in the clotting mechanism. Abciximab binds to GPIIb/IIIa receptor on both resting and activated platelets (~100,000 GPIIb/IIIa receptors per platelet cell), and inhibits platelet aggregation and resulting thrombus or clot formation without activating platelets. This selective inhibition of platelet aggregation is desirable because platelet adhesion, without aggregation and without interfering with the initial adhesion of platelets, may contribute to maintaining hemostasis.
The platelet GPIIb/IIIa receptor is a member of the integrin family of adhesion receptors and the major platelet surface receptor involved in platelet aggregation. Abciximab also binds with similar affinity to the vitronectin receptor, also known as the alpha integrin receptor. Abciximab does not interfere with GPIb-mediated platelet adhesion, and does not lead to measurable changes in platelet clearance (e.g., through the spleen). The mechanism of action of abciximab is thought to involve steric hindrance and/or conformational effects to block access of large molecules to the GPIIb/IIIa receptor, rather than direct interaction with the RGD (arginine-glycine-aspartic acid) binding site of GPIlb/IIIa. The precise binding site for abciximab is unknown.
GPIIb/IIIa receptor blockade and inhibition of platelet aggregation are highly correlated with 80% occupancy of receptors leading to nearly complete inhibition of platelet aggregation. Studies of the effects of abciximab in several well-defined animal models of thrombosis have shown that doses leading to more than 80% receptor blockade prevent arterial thrombosis. The vitronectin receptor mediates the procoagulant properties of platelets and the proliferative properties of vascular endothelial and smooth muscle cells. At concentrations in vitro providing >80% GPIIb/IIIa receptor blockage (but above the in vivo therapeutic range), abciximab more effectively blocks the burst of thrombin generation that follows platelet activation than other antibodies capable of inhibiting GPIlb/IIIa alone, indicating involvement of vitronectin receptors. Abciximab also binds to the vitronectin alpha(v)beta(3) receptor found on platelets and vessel wall endothelial and smooth muscle cells. The affinity of c7E3 for the GPIIb/IIIa receptor is Kd= 5 nM. The c7E3 monoclonal antibody Fab fragment (abciximab) binds to GPIIb/lIIa receptor on resting and activated platelets, inhibitsing platelet aggregation without activating platelets. Abciximab does not interfere with GPIb-mediated platelet adhesion and does not lead to measurable changes in platelet clearance (e.g., through the spleen).
Companies.: Reopro was developed by Centocor, Inc., subsequently acquired by Johnson & Johnson Co. (J&J). Reopro is manufactured by Centocor B.V. (Leiden, The Netherlands), CBER/FDA est. no. 1178, a subsidiary of Centocor, Inc./J&J. Centocor markets Reopro in the U.S. ReoPro is co-marketed in the U.S. by Centocor and Eli Lilly and Co., and Eli Lilly markets the product internationally. except in Japan, where it is marketed by Fujisawa Pharmaceuticals, Inc., Ltd. In spring 2005, Fujisawa completed its merger with Yamanouchi Pharmaceutical Co. Ltd.. forming Astellas Pharma Inc.
Abciximab has been reporred to be manufactured at Novartis facilities in Vacaville, CA, facilities originally belonging to Chiron. The facility includes a 20,000 L bioreactor.
However, the mAb has been reported to be manufactured using 5-20 bioreactors, 500 L each, at Leiden, operating in perfusion mode using internal spin-filters [Farid, S., Operational & Economic Evaluation of Integrated Continuous Biomanufacturing Strategies for Clinical & Commercial mAb Production, presented at ECI Integrated Continuous Biomanufacturing, Barcelona, Spain, 20-24 October 2013].
Eli Lilly & Co. at least partially funded and collaborated in development of ReoPro through a deal negotiated in July 1992 and renegotiated in June 1996, investing $124.5 million and $50.0 million in equity, and receiving U.S. co-marketing and international marketing rights, excluding Japan, and paying 50% royalties on ReoPro sales (according to Recombinant Capital).
Development of ReoPro was at least partially funded through Centocor Partners III, L.P. In Dec. 1999, Royalty Pharma, AG paid Centocor Partners III (or another limited partnership-type company that assisted in financing Reopro’s development) $27 million to acquire that organization’s future royalty interest in sales of ReoPro.
Manufacture: A vial of stored, frozen, transformed, myeloma cells from the Master Working Cell Bank (MWCB) is expanded in culture, and used to inoculate the production perfusion bioreactor(s). Continuous perfusion fermentation is performed in a defined medium, with the cells expressing the c7E3 (gamma 1 heavy chain, kappa light chain) monoclonal antibody. Recombinant expression includes use of technology (see Tech. transfer section) licensed from Celltech Biologics.
The Fab fragment of c7E3 (abciximab) is produced by enzymatic digestion of the purified chimeric 7E3 immunoglobulin with the proteolytic enzyme papain. The Fab fragment is isolated by a series of purification steps designed to yield a product free of other digestion fragments and other contaminating components (e.g., proteins, nucleic acids, viruses). Purification includes column chromatography, along with a robust virus inactivation step (unspecified), and a virus removal step (unspecified), perhaps referring to the papain digestion and column chromatography steps, respectively.
A battery of biochemical and functional assays has been developed to characterize the product during manufacture and afterwards. The functional assays (not specified) are somewhat unique for a biologic in that they very directly reflect the biological activity in the patient.
FDA class: Biologic PLA/ELA
CBER class: Blood And Blood Derivatives
CBER to CDER: Among the products transferred within FDA on June 30, 2003
Approvals: Date = 19941222; first approval, BLA; Indication = adjunct to percutaneous transluminal coronary angioplasty or atherectomy (PTCA) for prevention of acute cardiac ischemic complications in patients at high risk for abrupt closure of the treated coronary blood vessel
Date = 19971105, BLA supplement; Indication = expanded indications: to provide for revised dosage and patient management guidelines to reduce bleeding, and to include additional information on readministration. indications: included treatment of a broad range of patients undergoing percutaneous coronary intervention (PCI) and patients with unstable angina not responding to conventional medical therapy when PCI is planned within 24 hours.
Date = 19980218; BLA supplement; Indication = extension of clinical benefits through three years and additional clinical pharmacology data [including benefit seen on the primary endpoint of the EPIC trial and new in vitro data on the effects of abciximab on the vitronectin receptor and inhibition of thrombin generation]
Indications: [full text of "INDICATIONS AND USAGE” section from product insert/label]:
Abciximab is indicated as an adjunct to percutaneous coronary intervention for the prevention of cardiac ischemic complications in patients undergoing percutaneous coronary intervention in patients with unstable angina not responding to conventional medical therapy when percutaneous coronary intervention is planned within 24 hours.
Abciximab use in patients not undergoing percutaneous coronary intervention has not been studied.
Abciximab is intended for use with aspirin and heparin and has been studied only in that setting, as described in CLINICAL STUDIES.
Status: The original PLA/ELA was submitted in Dec. 1993, received priority review, and was approved on Dec. 22, 1994; review time = ~1.0 year. The product is now approved and marketed in most major markets worldwide.
The original approval was for bolus administration 10-60 minutes before PCI, followed by a 12-hour continuous infusion. The Nov. 1997 supplemental approval expanded use to patients undergoing percutaneous coronary intervention (PCI; balloon angioplasty, atherectomy, stent placement); and patients with refractory unstable angina could receive ReoPro 18-24 hours before PCI, concluding one hour after the procedure.
ReoPro received MAA approval in the European Union in May 1995.
Tech. transfer: The chimeric monoclonal antibody (c7E3) and its Fab fragment (abciximab) were exclusively licensed by Centocor from the State University of New York (SUNY). U.S. patent 5,770,198, “Platelet-specific chimeric 7E3 immunoglobulin,” issued June 23, 1998, coassigned to SUNY and Centocor, describes the 7E3 monoclonal antibody specific for glycoprotein IIb/IIIa receptor and recombinant chimeric fragments, including c7E3. The monoclonal antibody is produced by murine hybridoma ATCC acc. no. HB 8832. Plasmids p7E3VkappahCkappa and p7E3VH hCgamma4 carry the chimeric gene constructs encoding the light and heavy chains, respectively, of chimeric 7E3 immunoglobulin. Other U.S. patents covering aspects of abciximab and its uses include 5,656,272 (use for treatment of Crohn’s disease); 5,698,195 (use for treatment of rheumatoid arthritis); 5,877,006; and 5,919,452. See also WO 95 12412 and EP418316, assigned to Centocor and SUNY.
Lonza Biologics plc, a subsidiary of Alusuise-Lonza Group, receives unspecified royalties from Centocor/J&J on sales of ReoPro from licensing of its “Boss” patents covering use of the GS (glutamine synthetase) mammalian gene expression system. Related patents include U.S. 5,770,359 and 5,747,308. The technology is coassigned to the University of Glasgow (which presumably receives a share of royalties). This technology involves use of glutamine synthetase as a dominant selectable marker for co-amplification of genes and transforming host cell lines to glutamine independence. The glutamine synthetase gene is used in recombinant vectors as a marker along with the gene(s) for the desired protein, with only successfully transformed cells capable of producing their own GS and surviving in glutamine-deficient culture media. Over forty companies have licensed GS System technology for various uses.
Centocor/J&J has apparently nonexclusivley licensed the “Cabilly” recombinant chimeric/humanized monoclonal antibody design and expression patents, including U.S. 4,816,547, and related technology from Genentech, Inc. (and/or licensed related “Boss” patents from Celltech Group plc). See the Tech. transfer (rDNA) section of the Monoclonal Antibodies entry (#300) for further information about the complex patent, licensing, and patent disputes concerning recombinant chimeric/humanized monoclonal antibodies.
Currently, Genentech currently holds a dominant position in U.S recombinant chimeric/humanized monoclonal patents with its “Cabilly” patents (4,816,567 and 6,113,415), recent revocation a similar U.S. patent assigned to Celltech Group plc, and subsequent cross-licensing of the “Boss” patents held by Celltech. Genentech filed a patent infringement suit against Centocor alleging that ReoPro infringes its Cabilly patent in April 1994, and this was apparently resolved with Centocor taking a license from Genentech.
As discussed in the Recombinant DNA Product entry (at the beginning of this section), Genentech is appealing a $500+ million award to the City of Hope Medical Center (COH) arising from a 1976 contract and patent licensing dispute involving COH developing basic cloning technology for Genentech. Genentech is appealing to the California State Supreme Court.
Based on European patent EP418316, SPC certification (patent extension) was granted in the U.K. in Sept. 2003, effectively extending patent U.K. protection/exclusivity until Nov. 2009. It took nearly 13 years for Centocor/SUNY to obtain this patent, which is why it was received so long after the product’s approval.
Trials: Abciximab was approved based on three pivotal Phase III clinical trials evaluating the effects of ReoPro in patients undergoing percutaneous coronary intervention: in patients at high risk for abrupt closure of the treated coronary vessel (EPIC trial); in a broader group of patients (EPILOG trial); and in unstable angina patients not responding to conventional medical therapy (CAPTURE trial). Percutaneous intervention included balloon angioplasty (PCTA), atherectomy, or stent placement. All trials involved the use of various concomitant heparin dose regimens and, unless contraindicated, aspirin (325 mg) was administered orally two hours prior to the planned procedure and then once daily.
In its four Phase III clinical trials in more than 8,000 patients, ReoPro showed on average >50% improvement at 30 days in reducing the combined endpoint of death and heart attacks compared with placebo. In the EPISTENT trial, the combination of ReoPro and stents led to a 51% relative reduction in the composite primary endpoint of death, heart attack and urgent revascularization compared with stenting alone. This trial, along with the EPILOG, EPIC and CAPTURE trials, demonstrated that ReoPro provides a significant therapeutic benefit across a broad range of patients undergoing percutaneous coronary interventions.
In Oct. 2005, Lilly and J&J halted (permanently discontinued enrollment) in the double-blind, placebo-controlled, international, Phase III AbESTT-II (Abciximab in Emergent Stroke Treatment Trial-II) trial using Reopro for treatment of acute ischemic stroke due to instances of intracerebral hemorrhage and death. The trial had enrolled 811 of a planned 1,800 patients. The companies dropped further development for this indication, and planned to complete review and unblind the trial by March 2006.
Medical: Abciximab reduces complications associated with percutaneous transluminal coronary angioplasty (PCTA) or atherectomy by preventing the formation of blood clots that commonly intensify during the procedure.
Percutaneous transluminal coronary angioplasty (PTCA) is an effective method of enlarging the lumen of stenosed coronary arteries. There is an inherent risk of acute coronary occlusion during and after angioplasty, a major cause of in-hospital morbidity and mortality. Acute coronary occlusion during or immediately after coronary angioplasty appears to be caused by the combination of deep arterial wall injury with resultant occlusive thrombus formation.
Abrupt closure of newly opened arteries occurs in as many as l0-20% of high risk PTCA procedures, leading to death, myocardial infarction, or need for CABG or repeat PTCA. Certain patients are at particularly high risk for complications from angioplasty. These patients include those with certain angiographic lesion patterns (types B and C, defined by the ACC/AHA task force), age > 65, female sex, prior myocardial infarction, diabetes, prior CABG, impaired LVF, or a history of hypertension.
Platelet aggregation and subsequent clot formation (thrombosis) at the site of an atherosclerotic plaque is an important causative factor in the genesis of conditions such as angina, acute myocardial infarction, and reocculusion following successful thrombolysis (e.g., with thrombolytic agents) and angioplasty. Heart attack patients are typically treated with thrombolytic agents, such as a tissue plasminogen activator (tPA), which bread down the fibrin component of clots. A major complication associated with such fibrinolytic treatment is reocclusion based on platelet aggregation, which can result in further heart damage. Since glycoprotein (GP)IIb/IIIa receptors are known to be responsible for platelet aggregation, agents which block these receptors reduce or prevent reocclusion following thrombolytic therapy and accelerate the rate of thrombolysis. Abciximab binds to platelets, can block platelet aggregation, is a potent antithrombotic agent, and prevents or reduces reocclusion following thrombolysis.
Market: Total sales by Lilly were $231.5 million in 2009 and ~$208 million in 2008.
Total sales of ReoPro were estimated at ~$550 million in 2008; $363 million in 2004; $364.4 million in 2003; $384 million in 2002; $446 million in 1999; $365 million in 1998; $254 million in 1997; $149 million in 1996; and $23 million in 1995.
Analysts with Friedman Billings and Ramsey (FRB) have projected sales of $378 million in 2005, $376 million in 2006, and $378 million in 2007.
The 2007 Average Wholesale Price (AWP) is $660.14/5 mL, 2 mg/mL vial, with a Direct Price (discount price) of $550.12 (Red Book, 2007). The 2004 AWP was $584.02.
Fragmin (alteparin sodium; heparin fragments) from Pharmacia Corp./Pfizer Inc. joined ReoPro and Retavase (a recombinant thrombolytic agent) as the third product in Centocor’s (now J&J’s) cardiovascular portfolio in early 1999. Centocor/J&J and Pharmacia/Pfizer co-promote Fragmin. Together, ReoPro and Retavase command more than 50% of the acute coronary syndrome market.
Competition: ISU Abxis has received approval in South Korea for Clotinab, a biogeneric/biosimilar version of abciximab (ReoPro).
Companies involvement:
Full monograph
240 Platelet Mab Fab, rDNA
Nomenclature:
Platelet Mab Fab, rDNA [BIO]
ReoPro [TR (assigned to Eli Lilly & Co.)]
CenteoRx [TR former in U.S.]
Abciximab [FDA USAN INN BAN ]
Immunoglobulin G, (human-mouse monoclonal c7E3 clone p7E3VGHhhCgamma4) Fab fragment anti-human glycoprotein IIb/IIIa receptor), disulfide with human-mouse monoclonal c7E3 clone p7E3VkappahCkappa light chain [Note, the large H and gamma4 in “p7E3VGHhhCgamma4” and the kappa in “E3VkappahCkappa” are subscripted and gamma and kappa are Greek symbols]. [CAS]
143653-53-6 [CAS RN]
7E3, human-murine monoclonal antibody [SY]
anti-GPIIb/IIIa monoclonal antibody [SY]
c7E3 [SY]
c7E3 Fab fragment [SY]
Fab fragment of the chimeric human-murine monoclonal antibody 7E3 [SY]
GPIIb/IIIa monoclonal antibody [SY]
human-murine monoclonal antibody 7E3 [SY]
molecular weight (kDa) = 48
FDA Class: Biologic BLA
Year of approval (FDA) = 1994
Date of 1st FDA approval = 19941222
(in format YYYYMMDD)
Biosimilars/biobetters-related U.S. Patents: | 2015, based on 5,770,198.
Tech. Catalysts Intl., affiliated with Harvest Moon Pharm. reports 2017. |
U.S. Patent Expiration Year: | 2015 |
U.S. Biosimilars Data Exclusivity Expiration: | 2006 |
U.S. Biosimilars Orphan Exclusivity Expiration: | 2001 |
U.S. Biosimilars Launchability Year: | 2015 |
U.S. Biobetters Launchability Year: | 2015 |
Biosimilars/biobetters-related EU Patents: | 2012, for agent based on EP 0610201 and EP 1097945
for Crohn's disease use based on EP 0610201 2021, if EP 1309691 applies
2022, if EP 0418316 applies |
EU Patent Expiration Year: | 2012 |
EU Biosimilars Data Exclusivity Expiration: | 2005 |
EU Biosimilars Orphan Exclusivity Expiration: | 2005 |
EU Biosimilars Launchability Year: | 2012 |
EU Biobetters Launchability Year: | 2012 |
Index Terms:
antibodies (see also immune globulins; monoclonal antibodies)
biopharmaceutical products
bovine materials used<!-- bovinesource -->
exempt from CBER lot release requirements
monoclonal antibodies
monoclonal antibodies, recombinant
murine (mouse) materials used
receptor antibodies
recombinant DNA
ATCC HB8832
glutamine synthetase (GS) expression system
mammalian cell culture
murine (mouse) hybridoma cells
murine myeloma cells
p7E3VHhCg4
p7E3VkhCk
perfusion bioreactors
plasmid p7E3VkappahCkappa
rodent cells <!-- rodentcells -->
Sp2/0 murine hybridoma/myeloma cells
bovine source warning, unknown/undocumented country
calcium
cetyltrimethylammonium bromide (CTAB)
chelation
Factor XIII
papain digestion
polysorbate 80 (Tween 80)
sodium chloride
sodium phosphate
three-demensional cell culture
viral inactivation, unspecified
Water for Injection
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
catheter clearance
priority review status
EU200 Currently Approved in EU
UM001 Marketed Product in US
US200 Currently Approved in US
EM001 Marketed Product in EU
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