Desirudin - Iprivask; Revasc; desulfatohirudin, recombinant; hirudin (Hirudo medicinalis HV1), 1-L-leucine-2-L-threonine-63-desulfo-
Status: approved; marketed
Organizations involved:
Canyon Pharmaceuticals Inc. – US mark.
Sanofi Aventis Pharmaceuticals, Inc. – Former
Sanofi S.A. – Parent; Former
Novartis AG – Manuf.; Tech.
Madaus AG – Manuf. other
Ciba-Geigy Ltd. – Former
University of Washington – Tech
Genentech, Inc. – Tech
Washington Research Foundation (WRF) – Tech.
Cross ref.: See the Hirudin Products entry (#180) above, and the entry for Hirudin, rDNA/Berlex (#181). See also the entry for bivalirudin (Angiomax: #934), a fully synthetic hirudin peptide.
Description: Iprivask is a lyophilized (freeze-dried) formulation of recombinant desulfatohirudin polypeptide (desirudin) expressed by a transformed Saccharomyces cerevisiae (yeast; strain TR 1454) cell line, with post-expression enzymatic desulfation of the tyrosine at position 63 (Tyr-63) by an arylsulfatase enzyme. The amino acid sequence of desirudin is identical to that of natural hirudin except that it lacks a sulfate group on the tyrosine at position 63. Desirudin is composed of a single-chain (monomeric) of 65 amino acids with three disulfide bridges, with an isoeletric point of 3.8-4.3, a chemical formula of C287-H440-N80-O110-S6, and a molecular weight of 6963.52
The biological activity of desirudin is determined through a chromogenic assay which measures the ability of desirudin to inhibit the hydrolysis of a chromogenic peptidic substrate by thrombin in comparison to a desirudin standard. One vial of desirudin contains 15.75 mg desirudin corresponding to approximately 315,000 antithrombin units (ATU) or 20,000 ATU per milligram of desirudin with reference to the WHO International Standard (prepared 1991) for alphathrombin.
Desirudin has a specific activity of 18,000 antithrombin units (ATU) per mg of protein, referenced to the WHO Second International Standard for alpha-thrombin. The HPLC content is ~92%. One vial of desirudin contains 15.75 mg desirudin corresponding to approximately 315,000 antithrombin units (ATU) or 20,000 ATU/milligram of desirudin.
Iprivask 15 mg is packaged in single dose vials containing 15.75 mg desirudin, plus 1.31 mg anhydrous magnesium chloride USP, and sodium hydroxide for injection USP. Each ampule of diluent for Iprivask contains 0.6 mL sterile mannitol USP (3%) in Water for Injection (preservative free) for reconstitution and subcutaneous (SC) injection, preferably at an abdominal or thigh site. Iprivask is marketed in packages containing two and 10 vials each, along with diluent ampules. Iprivask is stored at 25ºC (77ºF), with excursions permitted to 15–30˚C (59-86˚F; room temperature).
Nomenclature: Hirudin, rDNA/Sanofi [BIO]; Iprivask [TR]; desirudin [INN; USAN]; Revasc [TR in Europe; reg. to Sanofi Pharma]; recombinant hirudin sequence variant 1 [EU]; hirudin (Hirudo medicinalis isoform HV1), 63-desulfo- [CAS]; 63-desulfohirudin [CAS]; 120993-53-5 [CAS RN]; desulfatohirudin, recombinant [SY]; CGP39393 [SY]; NDC 0075-2300-02; NDC 0075-2300-10 [NDC]
Note, some sources have mixed up nomenclature terms, particularly CAS names and numbers, for lepirudin (Refludan; see entry below) and Desirudin. The CAS RN for hirudin is 8001-27-2.
Companies.: Desirudin was originally developed and manufactured by Ciba-Geigy, now Novartis AG, at facilities in Basel, Switzerland, with finishing (filling, lyophilization, packaging, etc.) performed by Ciba-Geigy in Stein, Switzerland. About 1996-1997, finishing was moved to Dr. Madaus & Co., now Madaus AG (Cologne, Germany). With this, manufacturing batch size was increased from 40,000 to 60,000 vials.
In 1996, Ciba-Geigy AG merged with Sandoz AG to form Novartis AG. Both U.S. (FTC) and European Union regulators required divestment of the product as a condition for the merger. The companies signed a “Consent Agreement” to divest marketing of desirudin as a condition for the merger. Marketing rights were transferred to Aventis Pharmaceuticals (now Sanofi), which markets Revask internationally. Novartis AG and Madaus AG continue to manufacture desirudin for Sanofi Aventis S.A. Aventis Pharma merged into Sanofi Aventis S.A. in summer 2004. [Sanofi Aventis S.A. also manufactures lepirudin (Refludan) for marketing by Berlex/Schering and Pharmion (see lepirudin entry below)].
Canyon Pharmaceuticals Inc. assumed U.S. marekting in 2010.
Manufacture: Three different methods were used for manufacture of desirudin during its development. The two earlier methods provided low yields and the C-terminal end of the molecule was degraded by host cell carboxypeptidase during fermentation and purification. The commercial-scale method III using host strain S. cerevisiae TR 1454 provided improved yields, less degradation, and an improved impurity profile. Manufacturing methods, including the enzymatic desulfation enabling recombinant manufacture of desulfatohirudins, are further discussed in the “Tech. transfer” section below. Stable continuous constitutive expression of desulfatohirudin by Ciba AG (now Sandoz/Novartis) using S. cerevisiae strain 1456, apparently a pre-commercialization strain, was reported in Appl. Microb. Biotech., vol. 39, p. 526-31, 1993.
Desirudin is produced by S. cerevisiae strain 1454 transformed with plasmid pDP34/GAPFL-YHIR. The inserted sequence for desirudin was synthesized chemically using the known amino acid sequence of hirudin variant I (HVI) from the leech Hirudo medicinalis along with the signal sequence of yeast acid phosphatase (PH05). The expression of desirudin by yeast is controlled by the GAGPL promoter ligated to the coding sequence for the PH05 signal sequence. The resulting 1.1. kb expression cassette was cloned into the high copy number yeast shuttle vector pDP34. The signal sequence targets the expressed desirudin protein to the S. cerevisiae endoplasmic reticulum, where a peptidase enzyme cleaves the signal sequence, and desirudin is secreted into the culture medium.
Purification involves nine steps, including gel filtration to remove host cell proteins and other impurities of higher and lower molecular weight than desirudin. Purified desirudin is dissolved, pH adjusted to 7.4 with sodium hydroxide, sterile filtered, filled, lyophilized, and stoppers inserted and secured with aluminum caps. Magnesium chloride (stabilizer) is added after lyophilization). Vials are overfilled 5% to allow for withdrawal of 15 mg after reconstitution with 0.5 mL mannitol solution (15 mg mannitol in 0.5 mL Water for Injection)
Testing of each batch includes specifications for identity (description, SDS gel electrophoresis, amino acid sequence and peptide mapping), purity (isoelectric point, clarity, pH, water content and absorbance at 420 nm, sulfated ash, heavy metals, dimer and related substances, yeast proteins), DNA, microbial contamination and bacterial endotoxins), and potency (protein content and biological activity/mg).
A single yeast transformant was used for the master and working cell banks. Contamination is ruled out by repeated isolation of single yeast colonies and routine checking of the strain’s auxotrophic markers and specific protease deficiencies. A working cell bank of 50 vials stored in liquid nitrogen originates from 8 doublings relative to the master cell bank, with this considered sufficient to support production for 1-2 years [from European Product Assessment Report].
During its European Union (EU) review (~1996), meat (bovine-derived) peptone in the culture medium was replaced with soya (soybean plant-derived peptone). The resulting desirudin was accepted by the EU as comparable to that produced using the prior meat peptone-containing medium based on peptide mapping, microheterogenicity, amino acid sequencing, physio-chemical analysis, etc.
FDA class: Drug NDA
Approvals: Date = 20030404; NDA 21-271
Indications: [full text of the ‘"Indications and USAGE’ section of product insert/labeling; 4/2012]:
Iprivask is indicated for the prophylaxis of deep vein thrombosis, which may lead to pulmonary embolism, in patients undergoing elective hip replacement surgery.
Status: The NDA was filed on June 28, 2000 and approval was granted on April 4, 2003 (approval time = ~2.75 years). The first IND became effective on May 17, 1990.
Revasc was approved in the European Union on July 9, 1997 for the prevention of deep-vein thrombosis following hip and knee replacement surgery.
Note, Aventis and the companies which merged to form Aventis (now Sanofi Aventis S.A.) were previously involved in the development and marketing of lepirudin (Refludan), and Sanofi Aventis continues to manufacture lepirudin for marketing by Berlex/Schering and Pharmion (see Refludan entry).
Canyon Pharmaceuticals launched the product in the U.S. on March 2, 2010.
Tech. transfer: The FDA Orange Book reports patent expiration dates of May 17, 2005 for U.S. 4,745,177; Jan. 31, 2006 for U.S. 4,801,576; and March 31, 2015 for U.S. 5,733,874. In Europe, the SPC covering the product, based on EP 142860 (Germany), expired on Nov. 20, 2004.
U.S. 4,745,177, “Desulfatohirudins, the preparation thereof and pharmaceutical compositions containing them.” originally assigned to Ciba-Geigy Corp. (merged into Novartis) and Plantorgan Werk Heinrich G.E. Christensen KG (Bad Zwischenahn, Germany), now presumably transferred to Novartis AG, claims desulfatohirudins, including desirudin, irrespective of manufacturing methods (e.g., chemical synthesis, recombinant DNA). The inventors found that the “biological properties of hirudin are also retained if the characteristic sulfuric acid monoester group is removed from the phenolic hydroxyl group of the [Tyr63 ] residue.” They also note that the unusual hydroxysulfonyl (sulfated hydroxy) group in natural hirudins virtually rules out production by recombinant methods, while the absence of this group in desulfatohirudins enables this. Desulfatohirudins can be prepared by hydrolysis (enzymic desulfation) at Tyr-63 using arylsulfatase (e.g., Helix pomatia-derived enzyme from Boehringer Mannheim/Roche), which cleaves phenolic sulfate ester groups to form free phenolic groups under mild conditions.
U.S. 5,733,874, “Stable dry powders,” assigned to Novartis AG, claims lyophilized formulations of desulfatohirudin with water-soluble salts, e.g., magnesium chloride.
U.S. 4,801,576, “Method of inhibiting blood clotting with desulfatohirudins,” assigned to Ciba-Geigy Corp., now Novartis AG, claims methods for use of desulfatohirudins to inhibit clotting of mammalian blood.
U.S. 5,922,569, “Process for the production of polypeptides,” includes claims for use of CUP1 gene as a promoter for incorporation into two micron-derived hybrid plasmid vectors for expression of desulfatohirudin and a other proteins in protease-deficient transformed yeast.
Yeast expression system technology used for manufacture of lepirudin was developed and jointly patented by the University of Washington and Genentech. This has been nonexclusively licensed to Novartis AG by the Washington Research Foundation (WRF), the licensing agent for this family of yeast expression patents. Patents include U.S. 5,854,018 and 5,856,123 (continuations of 5,618,676) concerning recombinant production of proteins in yeast expression systems including Saccharomyces, Kluyveromyces, Pichia and Hansenula. These patents, particularly the more recent ones, broadly claim processes and materials for expression of proteins in yeast. All three U.S. patents will expire in 2014. The new patents are broader, as they claim processes and materials for expression of proteins in recombinant yeast systems general
Note, with both lepirudin and desirudin manufactured by expression in yeast by Sanofi Aventis, it is likely that some recombinant manufure-related patents related to these products originally assigned to one of the original developing companies, e.g., Ciba or Novartis, have now been (cross-) licensed to Sanofi Aventis.
As allowed for by the Drug Price Competition and Patent Term Restoration Act of 1984 (P.L. 98-417), the patent life of certain U.S. patents covering aspects of FDA-approved therapeutics can be extended (patent term restoration) based on time during development and regulatory review. The FDA determined that desirudin, exemplified by U.S. 4,745,177, had a total regulatory review period of 4,707 days, including 696 days during its testing phase, and 1,011 days during the approval/review phase. Patent extension time is based on complex calculations using these data. The applicants sought 5 years of patent term extension (the legal maximum), and this was presumably granted (for 4,745,177).
Trials: More than 2,000 patients were studied in two pivotal, controlled, randomized, multicenter, clinical efficacy trials, in addition to a controlled, double-blind, dose-finding study. Desirudin was compared to an unfractionated heparin and enoxaparin sodium for the reduction of the risk of venous thromboembolic events (VTE) in patients undergoing total hip replacement surgery. Iprivask was well tolerated in its controlled clinical trials, and treated patients had a lower incidence of VTE. The incidence of any hemorrhagic event was 30% with desirudin, 22% with heparin, and 33% with enoxaparin.
Two major studies involving a total of 1,564 patients showed desirudin 15 mg SC bd to be superior to unfractionated heparin 5000 iu tds in preventing deep vein thrombosis after elective hip surgery. Another study in 2,079 patients showed desirudin 15 mg SC bd to be more effective than enoxaparin (a synthetic lower molecular weight heparin) 40 mg SC daily in preventing deep-vein thrombosis after hip surgery. No differences in bleeding complications have been seen in the comparative studies between desirudin and the comparator agents. Thrombocytopenia, which can be associated with heparin and to a lesser extent with low molecular weight heparins, has not been reported after desirudin use.
Desirudin has also been studied in acute myocardial infarction, unstable angina and hemodialysis.
Disease: Deep vein thrombosis (DVT) is a significant and frequent complication of major orthopedic surgery, and patients undergoing such surgery of the pelvis, hip or lower limb are considered at “high risk” for DVT. The Thromboembolic Risk Factors (THRIFT) Consensus Group estimated the incidence of DVT (without prophylaxis) as 40-80% in this “high risk” group; proximal vein thrombosis risk at 10-30%; and fatal pulmonary embolism risk at 1-10%. Current DVT prophylactic regimens include low molecular weight heparin, or adjusted-dose or low-dose unfractionated heparin. Fixed dose standard heparin is known to have lesser efficacy in these high risk patients.
Medical: An initial dose is given 5-15 minutes prior to surgery, but after induction of regional block anesthesia, if used. Iprivasc is post-operatively administered as a 15 mg subcutaneous (SC) injection twice daily for 9-12 days, or until the patient is fully ambulant (whichever occurs first). The approved regimen (15 mg SC bd) prolongs activated prothrombin time (aPTT) to about 1.4 times of the baseline value.
Desirudin is absorbed completely following subcutaneous administration, with maximum plasma concentrations reached between 1-3 hours. Desirudin is metabolized and eliminated by the kidneys. The terminal elimination half-life is ~2 hours.
Desirudin is more effective for the prevention of thromboembolic events after orthopedic surgeries than regular or low molecular weight heparins, but has been associated with a similar incidence of hemorrhagic events.
Market: Average Wholesale Price (AWP) not available (inexplicably, the product is not in either the 2007, 2005 or 2004 Red Book).
The author’s rough guess for 2005 worldwide sales is in the range of ~$50-$100 million.
The cost for a course of treatment (15 mg 2x daily for 10 days) was reported by the National Health Service, U.K., upon approval (1997) to be £230.00, then 5-60 times the cost of low molecular weight and standard heparins.
Companies involvement:
Full monograph
182 Hirudin, rDNA/Sanofi
Nomenclature:
Hirudin, rDNA/Sanofi [BIO]
Iprivask [TR]
Revasc [TR]
recombinant hirudin sequence variant 1 [EU]
desirudin [INN]
63-Desulfohirudin (Hirudo medicinalis isoform HV1) [CAS]
hirudin (Hirudo medicinalis isoform HV1), 63-desulfo- [CAS]
[Leu1-Thr2]-63-desulfohirudin [CAS]
120993-53-5 [CAS RN]
CGP39393 [SY]
desulfatohirudin, recombinant [SY]
NDC 0075-2300-02; NDC 0075-2300-10 [NDC]
Note, some sources have mixed up nomenclature terms, particularly CAS names and numbers, for lepirudin (Refludan) and Desirudin (see entry below).
[[Notes]]
molecular weight (kDa) = 6.963
FDA Class: NDA Drug
Year of approval (FDA) = 2003
Date of 1st FDA approval = 20030404
(in format YYYYMMDD)
Biosimilars/biobetters-related U.S. Patents: | 2015, based on 5,733,874 (Orange Book); 2016, if 5,922,569, a process patent, or 5,733,874, a formulation patent, applies; 4,745,177 had been extended to 2010 |
U.S. Patent Expiration Year: | 2015 |
U.S. Biosimilars Data Exclusivity Expiration: | 2015 |
U.S. Biosimilars Orphan Exclusivity Expiration: | 2010 |
U.S. Biosimilars Launchability Year: | 2015 |
U.S. Biobetters Launchability Year: | 2015 |
Biosimilars/biobetters-related EU Patents: | 2015, based on EP 0665019, a formulation patent |
EU Patent Expiration Year: | 2015 |
EU Biosimilars Data Exclusivity Expiration: | 2007 |
EU Biosimilars Orphan Exclusivity Expiration: | 2007 |
EU Biosimilars Launchability Year: | 2015 |
EU Biobetters Launchability Year: | 2015 |
Index Terms:
biopharmaceutical products
bovine materials used<!-- bovinesource -->
exempt from CBER lot release requirements
recombinant DNA
yeast source materials
1-beta-D-ribofuranosyl-1,2,4-triazole-3-carboxamide
argon
Ga-23-902-530
heat treatment (pasteurization)
high-calcium medium
Hirudo medicinalis (medicinal leech)
leeches, medicinal
pCAVDHFRhuTNFRFc
pepsin digestion
peptides, synthetic
petrolatum gauze
plasmid pCAVDHFRhuTNFRFc
Saccharomyces cerevisiae (yeast)
Saccharomyces cerevisiae (yeast)
Souton medium
XV2181, Saccharomyces cerevisiae (yeast) strain
yeast KexII protease
lyophilized (freeze-dried)
Mab 1129, murine monoclonal antibody
mannitol
sodium hydroxide
Sterile Water for Injection
approval dates uncertain (FDA reports erroneous, conflicting, or simply has lost the original approval dates) (FDAapproved)
Park-William no. 8, Corynebacterium diphtheriae
EU200 Currently Approved in EU
UM001 Marketed Product in US
US200 Currently Approved in US
EM001 Marketed Product in EU
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