Aprotinin
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| Systematic (IUPAC) name |
| Aprotinin |
| Clinical data |
| AHFS/Drugs.com |
monograph |
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Pregnancy
category
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Legal status
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|
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Dependence
liability
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None |
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Routes of
administration
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Intravenous |
| Pharmacokinetic data |
| Bioavailability |
100% (intravenous) |
| Identifiers |
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CAS Registry Number
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9087-70-1 Y 9004-04-0 |
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ATC code
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B02AB01 |
| DrugBank |
DB06692 Y |
| UNII |
04XPW8C0FL Y |
| KEGG |
D02971 Y |
| ChEMBL |
CHEMBL1201619 N |
| Synonyms |
Trasylol, bovine pancreatic trypsin inhibitor |
| Chemical data |
| Formula |
C284H432N84O79S7 |
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Molecular mass
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6511.51 g/mol |
| N (what is this?) (verify) |
The drug aprotinin (Trasylol, previously Bayer and now Nordic Group pharmaceuticals), is the small protein bovine pancreatic trypsin inhibitor (BPTI), an antifibrinolytic molecule that inhibits trypsin and related proteolytic enzymes. Under the trade name Trasylol, aprotinin was used as a medication administered by injection to reduce bleeding during complex surgery, such as heart and liver surgery. Its main effect is the slowing down of fibrinolysis, the process that leads to the breakdown of blood clots. The aim in its use was to decrease the need for blood transfusions during surgery, as well as end-organ damage due to hypotension (low blood pressure) as a result of marked blood loss. The drug was temporarily withdrawn worldwide in 2007 after studies suggested that its use increased the risk of complications or death;[1] this was confirmed by follow-up studies. Trasylol sales were suspended in May 2008, except for very restricted research use. In February 2012 the European Medicines Agency (EMA) scientific committee reverted its previous standpoint regarding aprotinin, and has recommended that the suspension be lifted.[2] Nordic became distributor of aprotinin in 2012.[3]
Contents
- 1 Chemistry
- 2 Mechanism of drug action
- 3 Drug efficacy
- 4 Drug safety
- 5 In vitro use
- 6 History
- 7 Current findings
- 8 References
- 9 External links
Chemistry
| Pancreatic trypsin inhibitor |
|
| BPTI sequence, with its folded 3D structure represented by a ribbon for the secondary structure and a stick model (gray) for the backbone and sidechains. |
| Identifiers |
| Organism |
Bos taurus (domestic cow) |
| Symbol |
PTI |
| Entrez |
404172 |
| PDB |
4PTI More structures |
| RefSeq (mRNA) |
NM_001001554 |
| RefSeq (Prot) |
NP_001001554 |
| UniProt |
P00974 |
| Other data |
| Chromosome |
13: 75.02 - 75.03 Mb |
Aprotinin is a monomeric (single-chain) globular polypeptide derived from bovine lung tissue. It has a molecular weight of 6512 and consists of 16 different amino acid types arranged in a chain 58 residues long[4][5] that folds into a stable, compact tertiary structure of the 'small SS-rich" type, containing 3 disulfides, a twisted β-hairpin and a C-terminal α-helix.[6]
The amino acid sequence for bovine BPTI is RPDFC LEPPY TGPCK ARIIR YFYNA KAGLC QTFVY GGCRA KRNNF KSAED CMRTC GGA.[7] There are 10 positively charged lysine (K) and arginine (R) side chains and only 4 negative aspartate (D) and glutamates (E), making the protein strongly basic, which accounts for the basic in its name. (Because of the usual source organism, BPTI is sometimes referred to as bovine pancreatic trypsin inhibitor.)
The high stability of the molecule is due to the 3 disulfide bonds linking the 6 cysteine members of the chain (Cys5-Cys55, Cys14-Cys38 and Cys30-Cys51).[8] The long, basic lysine 15 side chain on the exposed loop (at top left in the image) binds very tightly in the specificity pocket at the active site of trypsin and inhibits its enzymatic action. BPTI is synthesized as a longer, precursor sequence, which folds up and then is cleaved into the mature sequence given above.
BPTI is the classic member of the protein family of Kunitz-type serine protease inhibitors. Its physiological functions include the protective inhibition of the major digestive enzyme trypsin when small amounts are produced, by cleavage of the trypsinogen precursor during storage in the pancreas.
Mechanism of drug action
Aprotinin inhibits several serine proteases, specifically trypsin, chymotrypsin and plasmin at a concentration of about 125,000 IU/ml, and kallikrein at 300,000 IU/ml.[5] Its action on kallikrein leads to the inhibition of the formation of factor XIIa. As a result, both the intrinsic pathway of coagulation and fibrinolysis are inhibited. Its action on plasmin independently slows fibrinolysis.[4]
Drug efficacy
In cardiac surgery with a high risk of significant blood loss, aprotinin significantly reduced bleeding, mortality and hospital stay.[5] Beneficial effects were also reported in high-risk orthopedic surgery.[5] In liver transplantation, initial reports of benefit were overshadowed by concerns about toxicity.[9]
In a meta-analysis performed in 2004, transfusion requirements decreased by 39% in coronary artery bypass graft (CABG) surgery.[10] In orthopedic surgery, a decrease of blood transfusions was likewise confirmed.[11]
Drug safety
There have been concerns about the safety of aprotinin.[5] Anaphylaxis (a severe allergic reaction) occurs at a rate of 1:200 in first-time use, but serology (measuring antibodies against aprotinin in the blood) is not carried out in practice to predict anaphylaxis risk because the correct interpretation of these tests is difficult.[5]
Thrombosis, presumably from overactive inhibition of the fibrinolytic system, may occur at a higher rate, but until 2006 there was limited evidence for this association.[5][10] Similarly, while biochemical measures of renal function were known to occasionally deteriorate, there was no evidence that this greatly influenced outcomes.[5] A study performed in cardiac surgery patients reported in 2006 showed that there was indeed a risk of acute renal failure, myocardial infarction and heart failure, as well as stroke and encephalopathy.[12] The study authors recommend older antifibrinolytics (such as tranexamic acid) in which these risks were not documented.[12] The same group updated their data in 2007 and demonstrated similar findings.[13]
In September 2006, Bayer A.G. was faulted by the FDA for not revealing during testimony the existence of a commissioned retrospective study of 67,000 patients, 30,000 of whom received aprotinin and the rest other anti-fibrinolytics. The study concluded aprotinin carried greater risks. The FDA was alerted to the study by one of the researchers involved. Although the FDA issued a statement of concern they did not change their recommendation that the drug may benefit certain subpopulations of patients.[14] In a Public Health Advisory Update dated October 3, 2006, the FDA recommended that "physicians consider limiting Trasylol use to those situations in which the clinical benefit of reduced blood loss is necessary to medical management and outweighs the potential risks" and carefully monitor patients.[15]
On October 25, 2007, the FDA issued a statement regarding the "Blood conservation using antifibrinolytics" (BART) randomized trial in a cardiac surgery population. The preliminary findings suggest that, compared to other antifibrinolytic drugs (epsilon-aminocaproic acid and tranexamic acid) aprotinin may increase the risk of death.[16] On October 29, 2006 the Food and Drug Administration issued a warning that aprotinin may have serious kidney and cardiovascular toxicity. The producer, Bayer, reported to the FDA that additional observation studies showed that it may increase the chance for death, serious kidney damage, congestive heart failure and strokes. FDA warned clinicians to consider limiting use to those situations where the clinical benefit of reduced blood loss is essential to medical management and outweighs the potential risks.[17] On November 5, 2007, Bayer announced that it was withdrawing Aprotinin because of a Canadian study that showed it increased the risk of death when used to prevent bleeding during heart surgery.[18]
Two studies published in early 2008, both comparing aprotinin with aminocaproic acid, found that mortality was increased by 32[19] and 64%,[20] respectively. One study found an increased risk in need for dialysis and revascularisation.[20]
No cases of bovine spongiform encephalopathy transmission by aprotinin have been reported, although the drug was withdrawn in Italy due to fears of this.[5]
In vitro use
Small amounts of aprotinin can be added to tubes of drawn blood to enable laboratory measurement of certain rapidly degraded proteins such as glucagon.
In cell biology aprotinin is used as an enzyme inhibitor to prevent protein degradation during lysis or homogenization of cells and tissues.
Aprotinin can be labelled with fluorescein isothiocyanate. The conjugate retains its antiproteolytic and carbohydrate-binding properties[21] and has been used as a fluorescent histochemical reagent for staining glycoconjugates (mucosubstances) that are rich in uronic or sialic acids.[22]
History
Initially named "kallikrein inactivator", aprotinin was first isolated from cow parotid glands in 1930.[23] and independently as a trypsin inhibitor from bovine pancreas in 1936.[24] It was purified from bovine lung in 1964.[25] As it inhibits pancreatic enzymes, it was initially used in the treatment for acute pancreatitis, in which destruction of the gland by its own enzymes is thought to be part of the pathogenesis.[26] Its use in major surgery commenced in the 1960s.[27]
BPTI is one of the most thoroughly studied proteins in terms of structural biology, experimental and computational dynamics, mutagenesis, and folding pathway. It was one of the earliest protein crystal structures solved, in 1970 in the laboratory of Robert Huber,[28] and was the first protein to have its structure determined by NMR spectroscopy, in the laboratory of Kurt Wuthrich at the ETH in Zurich in the early 1980s.[29][30]
Because it is a small, stable protein whose structure had been determined at high resolution by 1975,[31] it was the first macromolecule of scientific interest to be simulated using molecular dynamics computation, in 1977 by J. Andrew McCammon and Bruce Gelin, in the Karplus group at Harvard.[32] That study confirmed the then-surprising fact found in the NMR work[33] that even well-packed aromatic sidechains in the interior of a stable protein can flip over rather rapidly (microsecond to millisecond time scale). Rate constants were determined by NMR for the hydrogen exchange of individual peptide NH groups along the chain, ranging from too fast to measure on the most exposed surface to many months for the most buried hydrogen-bonded groups in the center of the β sheet, and those values also correlate fairly well with degree of motion seen in the dynamics simulations.
BPTI was important in the development of knowledge about the process of protein folding, the self-assembly of a polypeptide chain into a specific arrangement in 3D. The problem of achieving the correct pairings among the 6 Cys sidechains was shown to be especially difficult for the two buried, close-together SS near the BPTI chain termini, requiring a non-native intermediate for folding the mature sequence in vitro (it was later discovered that the precursor sequence folds more easily in vivo). BPTI was the cover image on a protein folding compendium volume by Thomas Creighton in 1992.[34]
Current findings
One scientific study in rats reported that treatment with aprotinin prevents disruption of the blood–brain barrier during the C. neoformans infection. [35]
References
- ^ "Bayer Temporarily Suspends Global Trasylol Marketing" (PDF) (Press release). Trasylol.com. 2007-11-05. Retrieved 2007-12-03.
- ^ "European Medicines Agency recommends lifting suspension of aprotinin". European Medicines Agency. 2012-02-17.
- ^ "The Nordic Group acquires rights to Trasylol® from Bayer HealthCare". The Nordic Group B.V. Retrieved 28 January 2014.
- ^ a b Mannucci PM; Mannucci, Pier Mannuccio (1998). "Hemostatic drugs". N. Engl. J. Med. 339 (4): 245–53. doi:10.1056/NEJM199807233390407. PMID 9673304.
- ^ a b c d e f g h i Mahdy AM, Webster NR (2004). "Perioperative systemic haemostatic agents". British journal of anaesthesia 93 (6): 842–58. doi:10.1093/bja/aeh227. PMID 15277296.
- ^ Richardson, J.S. (1981). "Anatomy and Taxonomy of Protein Structure". Advances in Protein Chemistry. Advances in Protein Chemistry 34: 167–339. doi:10.1016/S0065-3233(08)60520-3. ISBN 978-0-12-034234-1. PMID 7020376.
- ^ Kassell B, Radicevic M, Ansfield MJ, Laskowski M (1965). "The basic trypsin inhibitor of bovine pancreas. IV. The linear sequence of the 58 amino acids". Biochem. Biophys. Res. Commun. 18 (2): 255–8. doi:10.1016/0006-291X(65)90749-7. PMID 14282026.
- ^ Kassell B, Laskowski M (1965). "The basic trypsin inhibitor of bovine pancreas. V. The disulfide linkages". Biochem. Biophys. Res. Commun. 20 (4): 463–8. doi:10.1016/0006-291X(65)90601-7. PMID 5860161.
- ^ Xia VW, Steadman RH (2005). "Antifibrinolytics in orthotopic liver transplantation: current status and controversies". Liver Transpl. 11 (1): 10–8. doi:10.1002/lt.20275. PMID 15690531.
- ^ a b Sedrakyan A, Treasure T, Elefteriades JA (2004). "Effect of aprotinin on clinical outcomes in coronary artery bypass graft surgery: a systematic review and meta-analysis of randomized clinical trials". J. Thorac. Cardiovasc. Surg. 128 (3): 442–8. doi:10.1016/j.jtcvs.2004.03.041. PMID 15354106.
- ^ Shiga T, Wajima Z, Inoue T, Sakamoto A (2005). "Aprotinin in major orthopedic surgery: a systematic review of randomized controlled trials". Anesth. Analg. 101 (6): 1602–7. doi:10.1213/01.ANE.0000180767.50529.45. PMID 16301226.
- ^ a b Mangano DT, Tudor IC, Dietzel C (2006). "The risk associated with aprotinin in cardiac surgery". N. Engl. J. Med. 354 (4): 353–65. doi:10.1056/NEJMoa051379. PMID 16436767.
- ^ Mangano D, Miao Y, Vuylsteke A, Tudor I, Juneja R, Filipescu D, Hoeft A, Fontes M, Hillel Z, Ott E, Titov T, Dietzel C, Levin J (2007). "Mortality associated with aprotinin during 5 years following coronary artery bypass graft surgery". JAMA 297 (5): 471–9. doi:10.1001/jama.297.5.471. PMID 17284697.
- ^ Gardiner Harris (2006-09-30). "F.D.A. Says Bayer Failed to Reveal Drug Risk Study - New York Times". The New York Times. Retrieved 2007-11-05.
- ^ "Facts & Comparisons: Trasylol Public Health Advisory Update". Retrieved 2007-11-05.
- ^ U.S. Food and Drug Administration. "Early Communication about an Ongoing Safety Review Aprotinin Injection (marketed as Trasylol)". Archived from the original on 2007-10-30. Retrieved 2007-10-28.
- ^ U.S. Food and Drug Administration. "Information for Healthcare Professionals; Aprotinin (marketed as Trasylol)". Archived from the original on 2006-10-10. Retrieved 2006-10-30.
- ^ Gardiner Harris (2007-11-05). "Bayer Withdraws Heart Surgery Drug". The New York Times. Archived from the original on 7 November 2007. Retrieved 2007-11-05.
- ^ Shaw AD, Stafford-Smith M, White WD et al. (2008). "N Engl J Med". New England Journal of Medicine 358 (8): 784–793. doi:10.1056/NEJMoa0707768. PMID 18287601.
- ^ a b Schneewiss S, Seeger JD, Landon J, Walker AM (2008). "Aprotinin during coronary-artery bypass grafting and risk of death". N Engl J Med 358 (8): 771–783. doi:10.1056/NEJMoa0707571. PMID 18287600.
- ^ Stoddart RW, Kiernan, JA (1973). "Aprotinin, a carbohydrate-binding protein". Histochemie 34 (4): 275–280. doi:10.1007/BF00306299.
- ^ Kiernan JA, Stoddart RW (1973). "Fluorescent-labelled aprotinin: a new reagent for the histochemical detection of acid mucosubstances". Histochemie 34: 77–84.
- ^ Kraut H, Frey EK, Bauer E (1930). "Über die Inaktivierung des kallikreins". Hoppe-Seyler's Z Physiol Chem (in German) 192: 1–21. doi:10.1515/bchm2.1930.192.1-3.1.
- ^ Kunitz M, Northrup J (1936). "Isolation from beef pancreas of crystalline trypsinogen, trypsin, trypsin inhibitor, and an inhibitor trypsin compound". J Gen Physiol 19 (6): 991–1007. doi:10.1085/jgp.19.6.991. PMC 2141477. PMID 19872978.
- ^ Kraut H, Bhargava N (1964). "Versuche zur Isolierung des Kallikrein-Inaktivators aus Rinderlunge and seine Identifizierung mit dem Inaktivator aus Rinderparotis". Hoppe-Seyler's Z. Physiol. Chem. (in German) 338: 231–7. doi:10.1515/bchm2.1964.338.1-2.231. PMID 14330402.
- ^ Nugent FW, Warren KW, Jonasson H, Garciadeparedes G (1964). "Early experience with trasylol in the treatment of acute pancreatitis". South. Med. J. 57 (11): 1317–21. doi:10.1097/00007611-196411000-00012. PMID 14195953.
- ^ Tice DA, Worth Jr MH, Clauss RH, Reed GH (1964). "The inhibition of trasylol of fibrinolytic activity associated with cardiovascular operations". Surgery, gynecology & obstetrics 119: 71–4. PMID 14179354.
- ^ Huber, R. et al. (1970). "The Basic Trypsin Inhibitor of Bovine Pancreas. I. Structure Analysis and Conformation of the Polypeptide Chain". Naturwissenschaften 57 (8): 389–92. Bibcode:1970NW.....57..389H. doi:10.1007/BF00599976. PMID 5447861.
- ^ Wagner, G. and Wuthrich, K. (1982). "Sequential resonance assignments in protein 1H nuclear magnetic resonance spectra: basic pancreatic trypsin inhibitor". Journal of Molecular Biology 155 (3): 347–366. doi:10.1016/0022-2836(82)90009-2. PMID 6176717.
- ^ Havel, T.F. and Wuthrich, K. (1985). "An evaluation of the combined use of nuclear magnetic resonance and distance geometry for the determination of protein conformations in solution". Journal of Molecular Biology 182 (2): 281–294. doi:10.1016/0022-2836(85)90346-8. PMID 2582141.
- ^ Deisenhofer, J. and Steigemann, W. (1975). "Crystallographic Refinement of the Structure of Bovine Pancreatic Trypsin Inhibitor at 1.5 Angstroms Resolution". Acta Crystallographica B 31: 238. doi:10.1107/S0567740875002415.
- ^ McCammon JA, Gelin BR, Karplus M (1977). "Dynamics of folded proteins". Nature 267 (5612): 585–90. Bibcode:1977Natur.267..585M. doi:10.1038/267585a0. PMID 301613.
- ^ Wuthrich, K. and Wagner, G. (1975). "NMR investigations of the dynamics of the aromatic amino acid residues in the basic pancreatic trypsin inhibitor". FEBS Letters 50 (2): 265–268. doi:10.1016/0014-5793(75)80504-7. PMID 234403.
- ^ Thomas E. Creighton (1992). Protein Folding. W. H. Freeman. ISBN 978-0-7167-7027-5.
- ^ Xu CY (Feb 2014). "permeability of blood-brain barrier is mediated by serine protease during Cryptococcus meningitis.". J Int Med Res 42 (1): 85–92. doi:10.1177/0300060513504365. PMID 24398759.
External links
- The MEROPS online database for peptidases and their inhibitors: I02.001
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Antihemorrhagics (B02)
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Antihemorrhagics
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