Reteplase - Retavase; Repilysin; tissue plasminogen activator mutein, recombinant
Status - approved; marketed
Organizations involved:
Cornerstone Therapeutics Inc. – USA mark.
Chiesi Farmaceutici SpA – Parent
Actavis Group – Tech.; Intl. mark.
Fuji Diosynth Biotechnology – Manuf.
Hospira – Manuf. other
Scil Proteins GmbH – Tech.; Manuf.
Wacker GmbH – Parent
Protein Design Labs. – Tech.; Former
EKR Therapeutics Inc. – Former
ESP Pharma Inc. – Former
Centocor, Inc. – R&D; Tech.; Former
Johnson & Johnson Co. – Parent; Former
Boehringer Mannheim GmbH – Former
Hoffmann-La Roche Ltd. – Parent; Former
Covance Biotechnology Services Inc. – Former
Akzo Nobel NV – Parent
DuPont Pharmaceuticals – Former
Genentech, Inc. – Patent dispute
Royalty Pharma, AG. – Tech.
Cross ref.: See the Fibrinolytic and Thrombolytic Enzymes entry (#613), and the other tPA product entries below.
Description: Retavase (reteplase) is a lyophilized (freeze-dried) formulation of a recombinant mutein (modified form) of human tissue plasminogen activator (tPA) enzyme produced in transformed Escherichia coli (E. coli) bacteria. Retavase is considered a “third-generation” thrombolytic agent, genetically engineered to retain and delete certain portions of human tPA. Retavase is a deletion mutein of human tPA formed by deleting various amino acids present in endogenous human tPA. Retavase contains 355 of the 527 amino acids of native human tPA (amino acids 1-3 and 176-527), and retains the activity-related kringle-2 and serine protease domains of human tPA. Three domains are deleted from retavase – kringle-1, finger, and epidermal growth factor (EGF). These alternations enable rapid double-bolus injection and rapid initiation of thrombolysis (and associated reopening of blood clot-occluded coronary arteries). Retavase has a molecular weight of 39,571 Daltons (39.6 kDa) and a molecular formula of C1736-H2653-N499-O522-S22.
Nomenclature: tPA, rDNA/Centocor [BIO]; Retavase [TR]; reteplase [FDA INN]; 173-527-plasminogen activator (human tissue-type protein moiety reduced), 173-L-serine-174-L-tyrosine-175-L-glutamine- [CAS]; 133652-38-7 [CAS RN]; tissue plasminogen activator, recombinant [SY]; BM-06022 [SY]; fibrinokinase [SY]; Repilysin [TR used by Roche in Europe]; NDC 57894-040-01; NDC 57894-040-02 [NDC]
Biological.: See also other tPA product entries, particularly the following entry for alteplase (Activase), the first tPA product to enter the market.
Reteplase is a potent thrombolytic/fibrinolytic agent administered for the treatment of acute myocardial infarction (heart attack) to clear clots in coronary arteries and improve blood flow to heart muscle tissue. Potency is expressed in units (U) using a reference standard that is specific for reteplase/Retavase (not comparable with units used for any other thrombolytic agent).
Reteplase catalyzes degradation of endogenous plasminogen to active plasmin by cleaving a single peptide bond of plasminogen between the amino acids arginine and valine. The activation of plasminogen to plasmin is stimulated in the presence of fibrin (the primary structural component of blood clots), and is mediated via the tPA (and retavase) kringle-2 domain. Activated plasmin enzymatically degrades the fibrin matrix of thrombi (blood clots), exerting potent thrombolytic/fibrinolytic (“clot busting”)action.
The deletion of the three domains from human tPA and the lack of glycosylation (oligosaccharide side chains; due to bacterial expression) of reteplase results in faster plasma clearance and shorter half-life (about 11-19 minutes), compared to unmodified tPA, both endogenous and recombinant forms (e.g., alteplase or Activase, from Genentech). Compared to tPA, reteplase does not bind fibrin as tightly. This allows it to diffuse more freely through the clot, rather than binding primarily to the fibrin clot surface as tPA does. Reteplase in high concentrations does not compete with plasminogen for fibrin-binding sites, allowing plasminogen at the clot to be transformed into clot-dissolving plasmin.
The modifications incorporated into reteplase contribute to faster reperfusion in treated patients, compared alteplase or other unmodified recombinant tPA. The shorter half-life of reteplase allows double-bolus dosing– 10 U by intravenous injection in less than 2 minutes, followed 30 minutes later by a second 10 U intravenous bolus. This regimen offers more convenient administration and faster thrombolysis compared to recombinant tPA (alteplase/Activase), which is generally given by a 15 mg bolus followed by a 90-minute IV infusion.
Retavase is packaged as a preservative-free powder in vials containing 10.4 U (18.10 mg) reteplase to ensure administration of 10 U. Each vial is part of a kit with components for reconstitution and administration. Each single-use vial also contains tranexamic acid, 8.32 mg; dipotassium hydrogen phosphate, 136.24 mg; phosphoric acid, 51.27 mg; sucrose 364.00 mg; and polysorbate 80 5.20 mg.
Retavase is supplied in a kit (NDC 57894-040-01) containing two single-use vials of reteplase and two single-use 10 mL diluent vials, with needles and syringes. After reconstitution wit Sterile Water for Injection, USP (without preservatives), provided as part of the kit, Retavase has a pH of 6.0 ± 0.3. Kits also contain two sterile 10 mL syringes with 20 gauge needles attached, two sterile dispensing pins, and two alcohol swabs. In May 2000, Centocor introduced the first shielded needle for a fibrinolytic product, packaged with Retavase kits. The needle has a plastic sheath that covers the needle upon withdrawal from an intravenous line, providing protection from needle sticks.
The dating period for Reteplase is 24 months from the date of manufacture when stored at 2-25˚C. The date of manufacture is the date of the first sterile filtration of final formulated bulk. The dating period for the administration kit is 24 months or less, based on the shortest expiration date of any component in the kit, when stored at 2-25˚C.
Companies.: In Feb. 2005, ESP Pharma Inc. acquired U.S. and Canadian rights to Retavase reteplase from Centocor/J&J. Protein Design Labs. (PDL) announced its plans to acquire EPS in Jan. 2005, and completed its acquisition in March 2005. PDL then marketed Retavase in the U.S. In Feb. 2008, EKR Therapeutics acquired PDL’s cardiovascular assets, primarily Retavase, for $85 million and potentially up to $85 million more in development and sales milestones. International marketing was previously licensed by PDL to Hoffmann-La Roche AG.
PDL continued to manufactures Retavase (according to the online product insert online, 9/9/2008). However, the EKR Therapeutics Web site had reported, "Retavase® is manufactured for EKR Therapeutics Inc. at Hospira, Inc., McPherson, KS," but with this apparently referring to product filling, finishing, packaging, etc.
Retavase/Rapilysin was originally developed and manufactured by and the original FDA approval was granted to Boehringer Mannheim GmbH (BM); CBER/FDA est. no. 1211. BM manufactured reteplase at its facility in Penzberg, Germany, and formulated, filled, lyophilized and labeled the product at its facility in Mannheim, Germany. DuPont Pharmaceuticals Inc. and Boehringer Mannheim Corp. (U.S. subsidiary) co-promoted Retavase in the U.S. BM marketed the product in Europe and internationally.
In late 1997, the privately-held Corange Group, the owner of Boehringer Mannheim, agreed to be acquired by Roche Holding Ltd. (F. Hoffmann-La Roche Ltd.). Due to antitrust (monopoly) concerns, the Federal Trade Commission (FTC) required divestment of Retavase U.S. marketing rights as a condition to approving to the merger. Roche already held controlling interest in Genentech, which then marketed Activase, another form of tPA, for similar indications: (and now also markets its own tPA mutein, TNKase).
Centocor, Inc. (now part of Johnson & Johnson) concluded an agreement with F. Hoffmann-La Roche Ltd. in March 1998 to acquire U.S. and Canadian rights to Retavase for $335 million, with Roche retaining European and other international marketing rights. Roche continued to market the product in Europe under the trade name Repilysin. On Dec. 1, 1998, Centocor withdrew U.S. co-promotion of Retavase from DuPont Pharmaceuticals, leaving Centocor/J&J with exclusive U.S. marketing rights since Jan. 1, 1999. DuPont continued to receive unspecified royalties based on sales in its previous territories.
In April 1999, Centocor announced a long-term agreement with Covance Biotechnology Services Inc., now Fuji Diosynth Biotechnology, for the contract manufacture of Retavase. Under the agreement, Covance (now Fuji Diosynth) exclusively manufactured reteplase (at least for the U.S. and Canada), most likely at its facilities in Oss, The Netherlands. Additional terms were not disclosed.
In July 1999, Johnson & Johnson Co. acquired Centocor, Inc.
Royalty Pharma, AG has purchased rights to certain royalties from sales of Retavase.
On Jan. 25, 2005, PDL announced it would acquire privately held ESP Pharma for about $489 million. In Feb. 2005, ESP Pharma agreed to acquire from Centocor/J&J rights to manufacture, develop, market and distribute Retavase in the U.S. and Canada for $110 million and milestone payments of up to $45 million. PDL then agreed to increase the purchase price paid to ESP Pharma’s shareholders by $25 million, and agreed to assume ESP’s purchase obligations to Centocor.
In 2006, Covance/Diosynth encountered some problems in the manufacture of Retavase.
In Feb. 2007, PDL BioPharma extended its contract with Diosynth for exclusive manufacture of reteplase.
In Aug. 2007, PDL Biopharma announced plans to sell off its marketed products, including Retavase, to concentrate on antibody discovery and development. PDL hired Merrill Lynch & Co. to assist it in finding purchasers for its products. In Feb. 2008, EKR Therapeutics acquired Retavase (U.S. marketing rights).
In Aug. 2007, Actavis acquired exclusive European and worldwide distribution rights for Rapilysin, excluding USA and Canada, from Roche and manufacturing technology from PDL.
In Jan. 2012, Actavis concluded an agreement with Scil Proteins for contract manufacture of reteplase/Rapilysin (for Actavis' international markets). Scil Proteins had originally signed an agreement with Roche in June 2008 for process transfer and manufacture of Reteplase from its Penzberg facilities to Scil in Halle, citing this as a "demanding production process requiring considerable experience in protein refolding that only Scil Proteins could deliver."
In May 2012, Cornerstone Therapeutics Inc. (majority-owned by Chiesi Farmaceutici, SpA) acquired EKR Therapeutics. Cornerstone reported, " In 2013, Cornerstone is targeting FDA approval of a new
active ingredient supplier [Scil] and relaunch of RETAVASE, which could increase
revenues significantly versus 2012."
In Oct. 2012, Scil Proteins' facility in Halle, Germany, received EMA/EU approval for manufacture of reteplase. FDA facility approval was expected in 2013.
In Nov. 2013, Wacker GmbH acquired Scil Proteins.
Manufacture: As manufactured by Boehringer Mannheim (and presumably continued), reteplase is produced by fermentation of plasmid-transformed E. coli K12 bacteria. The coding sequence for reteplase was integrated into an expression vector (plasmid) and the E. coli cell line was transformed along with a helper vector (plasmid) which improves the yield of reteplase and reduces the risk of translation errors. The plasmids were introduced into E. coli by the calcium chloride method. Transformed E. coli cell banks, the Master Cell Bank (MCB), and the manufacturer’s Working Cell Bank (MWCB), have been verified and tested for genetic markers, antibiotic resistance patterns, restriction enzyme analysis, and expression of reteplase. The absence of bacteriophages is demonstrated using a plaque assay.
Fermentation takes place in 1,000 Liter production fermentor(s). A vial of MWCB is successively cultured and expanded into 100 mL and 10 L cultures (both containing ampicillin and kenamycin antibiotics), and this is used as inoculum for the production fermentor. The E. coli is cultured, harvested, cells are disrupted, inclusion bodies containing inactive reteplase are isolated, the protein is converted into its active form by an in vitro folding process, and active reteplase is purified by chromatographic separation.
Refolding is critical to transform the expressed protein into the active enzyme. Under reducing conditions, the cysteine residues or reteplase are modified by glutathione (reducing agent), resulting in a mixed disulfide solution. Inclusion bodies are solubilized and denatured under reducing conditions, and glutathione reducing/refolding agent is removed. This is followed by four purification steps to concentrate retavase and remove incorrectly folded molecules – acidification plus filtration; affinity chromatography on Erythrina trypsin inhibitor (ETI)-Sepharose; and two ion-exchange chromatography steps. This is followed by concentration, diafiltration, and sterile filtration. The active ingredient, reteplase, in an arginine buffer is the bulk drug substance. The manufacturing process has been validated to assure consistency of yield, degree of purity, and quality of the active ingredient. Potency is determined by amidolytic activity and clot lysis assays.
FDA class: Biologic BLA
CBER class: Blood And Blood Derivatives
CBER to CDER: Among the products transferred within FDA on June 30, 2003
Approvals: Date = 19961030, BLA ref. no. 95-1142 and 95-1167; granted to Boehringer Mannheim GmbH
Date = 19980506; approval revoked and granted (reissued) to Centocor, Inc. (est. no. 1242)
Indications: [full text of the "INDICATIONS AND USAGE” section of U.S. product insert/labeling]:
Retavase (Reteplase) is indicated for use in the management of acute myocardial infarction (AMI) in adults for the improvement of ventricular function following AMI, the reduction of the incidence of congestive heart failure and the reduction of mortality associated with AMI. Treatment should be initiated as soon as possible after the onset of AMI symptoms (see CLINICAL PHARMACOLOGY).
Indications: [full text of the "Therapeutic indications" section of the EU SPC]
Status: Retavase is exempt from CBER, FDA, requirements for lot release approval prior to marketing.
On Aug. 29, 1996, European Union approval (MAA) for Ecokinase (Reteplase) was granted to Galenous Mannheim (Boehringer Mannheim), which was later acquired and merged into Hoffmann-La Roche Ltd.
Tech. transfer: Boehringer Mannheim GmbH has received patents related to reteplase including EP 0242836, “Tissue Plasminogen Activator (tPA) Derivative and Its Production,” by Mattes, R., filed April 21, 1987. The patent covers the design and preparation of tPA deletion muteins. No reteplase-specific granted U.S. patents assigned to Centocor or Boehringer Mannheim were readily identified.
In March 1998, Genentech, Inc. filed suit in U.S. District court alleging that Centocor was infringing its U.S. patents, 5,728,565 and 5,728,566, concerning methods for production of recombinant tPA. In Feb. 1999, Genentech filed another suit in federal court alleging that Centocor was infringing its recently issued U.S. patent 5,869,314 relating to variant forms of t-PA, particularly t-PA deletion variants having characteristics different from another thrombolytic agent, urokinase. The patent’s claims include amino acid deletion derivatives and amino acid substitution derivatives. This dispute was apparently settled out-of-court, and without Centocor making substantial continuing royalty payments to Genentech.
Market: Total worldwide sales were $276.9 million in 2005, and $96.0 million in 2004. Total Retavase sales for 1999 (1st two quarters only; prior to merger with J&J) reported by Centocor were $44 million (rate equiv. to about $88 million/year). Total 1998 sales were $65.2 million including $55.4 million sales reported by Centocor (with the other sales by Boehringer Mannheim). Total 1997 sales by Boehringer Mannheim were $44.8 million.
Net product sales for the 12 months ended Sept. 30, 2007, were $21.6 million. Total 2006 sales reported by PDL Biopharma were $30.8 million. This decrease in sales may have been due to a manufacturing-related shortage of product.
With the 2005 acquisition of U.S. rights to Retavase by PDL, the company reported it anticipates compound annual growth rates of ~25% for net product sales of Retavase for each year from 2006 through 2008.
The 2007 Average Wholesale Price (AWP) for Retavase is $1,436.25/i.v. kit with one 18.1 mg vial (Red Book, 2007), unchanged from 2004, and $2,895.48/two 18.1 mg vials.
In Canada. Retavase is reported to cost $1,900.00/treatment (2 x 10.4U vials).
Retavase is used predominantly by cardiologists, cardiothoracic surgeons and emergency room physicians for treatment of acute myocardial infarction (AMI). Retavase, offering benefits over prior products, was launched in the U.S. in Jan. 1997, and has captured significant share of the market. However, it now has competition from another fast-acting t-PA mutein, TNKase, from Genentech.
Ongoing: The ongoing (2005) FINESSE trial was being conducted by Centocor (now, presumably, PDL) and Eli Lilly to study Retavase in combination with Reopro (abciximab) in the setting of facilitated percutaneous coronary intervention (PCI).
Companies involvement:
Full monograph
268 tPA, rDNA/PDL
Rapilysin is indicated for the thrombolytic treatment of suspected myocardial infarction with persistent ST
elevation or recent left Bundle Branch Block within 12 hours after the onset of acute myocardial infarction
AMI symptoms.
Nomenclature:
tPA, rDNA/PDL [BIO]
Retavase [TR]
Reteplase [FDA INN]
173-527-plasminogen activator (human tissue-type protein moiety reduced), 173-L-serine-174-L-tyrosine-175-L-glutamine- [CAS]
133652-38-7 [CAS RN]
BM-06022 [SY]
fibrinokinase [SY]
t-PA [SY]
Tissue plasminogen activator, recombinant [SY]
Rapilysin [TR former used by Roche in Europe]
Ecokinase [TR former used by Galenus Mannheim, Germany
]
NDC 57894-040-01; NDC 57894-040-02 [NUM NDC]
C1736H2653N499O522S22 [MF USAN]
39.57189 kDa [MW USAN]
molecular weight (kDa) = 39.6
FDA Class: Biologic BLA
Year of approval (FDA) = 1996
Date of 1st FDA approval = 19961030
(in format YYYYMMDD)
Biosimilars/biobetters-related U.S. Patents: | 2015, based on 5,728,565 and 5,728,566, cross-licensed by Genentech from J&J |
U.S. Patent Expiration Year: | 2015 |
U.S. Biosimilars Data Exclusivity Expiration: | 2008 |
U.S. Biosimilars Orphan Exclusivity Expiration: | 2003 |
U.S. Biosimilars Launchability Year: | 2015 |
U.S. Biobetters Launchability Year: | 2015 |
Biosimilars/biobetters-related EU Patents: | 2013, based on EP 0093619 |
EU Patent Expiration Year: | 2013 |
EU Biosimilars Data Exclusivity Expiration: | 2006 |
EU Biosimilars Orphan Exclusivity Expiration: | 2006 |
EU Biosimilars Launchability Year: | 2013 |
EU Biobetters Launchability Year: | 2013 |
Index Terms:
biopharmaceutical products
enzymes
enzymes, fibrinolytic/thrombolytic
exempt from CBER lot release requirements
recombinant DNA
bacterial culture <!-- bacterialculture -->
Escherichia coli (E. coli)
fermenters, 1,000 liter
K-12, Escherichia coli (E. coli)
amidolysis assays
arginine
calcium chloride
dipotassium hydrogen phosphate
disulfide solution
Erythrina trypsin inhiibitor (ETI)-Sepharose
fibrin
lyophilized (freeze-dried)
phosphoric acid
plasmin
plasminogen
polysorbate 80 (Tween 80)
Retavase reference standard
Sepharose, Erythrina trypsin inhibitor (ETI)-
Sterile Water for Injection
sucrose
tranexamic acid
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
catheter clearance
exempt from CBER lot release requirements
EU200 Currently Approved in EU
UM001 Marketed Product in US
US200 Currently Approved in US
EM001 Marketed Product in EU
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