pe
pep-04443 v1 CC-BY-SA-4.0

Aprotinin (Trasylol): cuts bleeding during heart surgery

A cow-derived protein dripped into a vein during open-heart surgery to reduce blood loss; pulled from the market in 2007 over safety concerns and later allowed back in Europe for limited use, check current status in your country.

statusbioassayed target? length58 aa refs23
fda-approved
status 5 / 5
sequence58 aa
151015202530354045505558
RPDFCLEPPYTGPCKARIIR YFYNAKAGLCQTFVYGGCRA KRNNFKSAEDCMRTCGGA
overview readme

What this is

Aprotinin (sold as Trasylol) is a small bovine protein, given by intravenous infusion to reduce surgical bleeding — primarily in adults undergoing coronary artery bypass graft (CABG) surgery. It is a 58-amino-acid serine protease inhibitor originally purified from cow tissue, and it works by blocking the body's bleeding-promoting enzymes during the artificial-circulation phase of heart surgery. The drug has had a turbulent regulatory history: widely used from the late 1980s onward, withdrawn worldwide in 2007–2008 over safety concerns, and reinstated in the European Union in 2012–2013 for a restricted indication after the data behind the withdrawal were re-evaluated.

The stored 58-residue sequence is the mature bovine pancreatic trypsin inhibitor (BPTI) backbone. Aprotinin is a Kunitz-type fold stabilised by three disulfide bonds (Cys5–Cys55, Cys14–Cys38, Cys30–Cys51 in the canonical BPTI numbering); those bridges are not visible in the linear single-letter sequence but are essential to the inhibitor's compact pear-shaped structure and its ability to engage active-site serines.

History

Aprotinin was first identified in the 1930s as a bovine inhibitor of trypsin- and kallikrein-like enzymes, and the protein was thoroughly characterised through the 1950s–1970s as bovine pancreatic trypsin inhibitor (BPTI). It became one of the most intensively studied small proteins in structural biology — one of the earliest protein crystal structures solved — and is the archetypal member of the Kunitz protease-inhibitor family.

The drug's modern clinical career began with a landmark 1987 trial: Royston and colleagues at the Royal Brompton Hospital reported that high-dose aprotinin reduced mean blood loss from about 1,500 mL to under 300 mL in 22 patients undergoing repeat open-heart surgery, with parallel reductions in transfusion requirement (Royston et al., The Lancet, 1987). That trial transformed aprotinin (marketed by Bayer as Trasylol) into a routine intra-operative agent in cardiac surgery worldwide over the next two decades, and a 1987 mechanistic paper documented its effects on the hemostatic disturbances of cardiopulmonary bypass (van Oeveren et al., Annals of Thoracic Surgery, 1987).

The story turned in 2006 when Mangano and colleagues published an observational analysis in The New England Journal of Medicine reporting that aprotinin use was associated with renal failure, myocardial events, stroke and encephalopathy compared with lysine antifibrinolytics or no antifibrinolytic (Mangano et al., NEJM, 2006), followed by a 2007 JAMA study reporting increased five-year mortality after CABG in aprotinin-exposed patients (Mangano, JAMA, 2007). The Canadian BART trial — a randomized comparison of aprotinin against tranexamic acid and aminocaproic acid in high-risk cardiac surgery — was halted early in 2008 after an interim analysis showed a higher 30-day mortality rate in the aprotinin arm (6.0% vs ~4%) despite a modest bleeding advantage (Fergusson et al., NEJM, 2008; commentary Ray, NEJM 2008). Bayer suspended marketing worldwide that year.

A subsequent re-analysis judged the BART results unreliable; the European Medicines Agency lifted its suspension in February 2012, and the European Commission formally reinstated marketing authorisation in 2013 for a restricted indication. Royston's 2014 review in Anaesthesia summarised the post-reinstatement evidence and the renewed clinical case for aprotinin in selected patients (Royston, Anaesthesia, 2014).

What it does

In the context of heart surgery on cardiopulmonary bypass, aprotinin blunts the surge of clot-dissolving and inflammatory enzyme activity that the artificial circuit triggers, and the result is less postoperative bleeding and a lower need for donor-blood transfusion. The original 1987 trial showed roughly a five-fold reduction in mean blood loss in repeat heart surgery (Royston 1987); subsequent work extended this to specific high-bleeding-risk populations including patients on clopidogrel presenting for CABG (van der Linden et al., Circulation, 2005) and to profoundly hypothermic perfusion (Westaby et al., European Journal of Cardio-Thoracic Surgery, 1994).

Beyond the antifibrinolytic effect, aprotinin has documented anti-inflammatory actions on the bypass-induced systemic response — Royston reviewed the role of aprotinin and other protease inhibitors in preventing the inflammatory response to open-heart surgery (Royston, International Journal of Cardiology, 1996) — and it preserves coronary bypass-graft vascular reactivity in animal models of cardiac surgery (Allen et al., Journal of Thoracic and Cardiovascular Surgery, 1997).

Mechanism

Aprotinin is a broad-spectrum serine protease inhibitor of the Kunitz family. Its compact, disulfide-stabilised fold presents a "canonical loop" that occupies the active site of target serine proteases as a pseudo-substrate, blocking catalysis. Clinically the most relevant targets are plasmin (the fibrinolytic enzyme that breaks down clots) and plasma kallikrein (the contact-system enzyme that drives kinin generation and contributes to the inflammatory response to cardiopulmonary bypass). The 2014 Royston review draws a sharp mechanistic contrast with the lysine analogues tranexamic acid and ε-aminocaproic acid: those drugs prevent plasminogen from binding fibrin (so they block plasmin formation), whereas aprotinin inactivates free plasmin that has already been generated — a different point of intervention in the same cascade (Royston, Anaesthesia, 2014).

Pharmacokinetically, aprotinin is administered intravenously during surgery; a weight-related dosing regimen has been evaluated for producing more predictable and constant plasma concentrations than fixed dosing (Royston et al., Anesthesia & Analgesia, 2001).

Evidence

  • Human (efficacy on bleeding and transfusion): Multiple randomized and observational studies established that aprotinin reduces perioperative blood loss and transfusion need in cardiac surgery — the original Royston 1987 Lancet trial in repeat open-heart surgery; the van der Linden 2005 Circulation study in clopidogrel-treated CABG patients; the Westaby 1994 trial in hypothermic perfusion; and the BART trial itself confirmed a reduction in massive bleeding (74/781 aprotinin vs 93–94/~770 in each lysine-analogue arm) (Fergusson et al., NEJM, 2008).
  • Human (safety controversy): Mangano's 2006 NEJM observational analysis and 2007 JAMA five-year mortality follow-up reported safety signals against aprotinin in CABG; BART showed a 30-day mortality difference; Henry and colleagues' 2009 CMAJ meta-analysis of aprotinin versus lysine-derived antifibrinolytics quantified the comparative safety profile (Henry et al., CMAJ, 2009); Takagi and colleagues' meta-analysis of head-to-head randomized trials reported increased mortality with aprotinin compared with tranexamic acid (Takagi et al., Interactive CardioVascular and Thoracic Surgery, 2009). Re-analyses and editorial responses contested these conclusions (Bizouarn, Annales Françaises d'Anesthésie et de Réanimation, 2006; Bremerich et al., Der Anaesthesist, 2006; Immer et al., Heart Surgery Forum, 2008; Augoustides, Drug Safety, 2008; Pagano et al., Journal of Thoracic and Cardiovascular Surgery, 2008), and the EMA review that underpinned reinstatement concluded the BART results were unreliable.
  • Cognitive outcomes: Postoperative cognitive dysfunction in aprotinin-exposed cardiac surgery patients has been examined as a separate question from mortality (Murkin, Canadian Journal of Anesthesia, 2004).

Known effects

  • Reduced perioperative blood loss in cardiac surgery — consistently demonstrated since Royston 1987; mechanism is plasmin and kallikrein inhibition.
  • Reduced allogeneic blood transfusion requirement in CABG — including in patients on antiplatelet therapy (van der Linden 2005).
  • Modulation of the systemic inflammatory response to cardiopulmonary bypass — protease-inhibition–mediated, reviewed by Royston 1996.

Safety signals

The published safety record is the central element of aprotinin's regulatory history. Mangano and colleagues' observational NEJM 2006 study reported increased rates of renal dysfunction, myocardial infarction and cerebrovascular events in aprotinin-treated CABG patients compared with controls (Mangano et al., NEJM, 2006), and Mangano's JAMA 2007 follow-up reported a higher five-year all-cause mortality in the aprotinin-exposed cohort (Mangano, JAMA, 2007). The BART randomized trial reported a 30-day all-cause mortality of 6.0% in the aprotinin arm versus 3.9% with tranexamic acid and 4.0% with aminocaproic acid (Fergusson et al., NEJM, 2008). Henry and colleagues' CMAJ 2009 meta-analysis and the Takagi 2009 head-to-head meta-analysis quantified comparative risk relative to lysine analogues (Henry et al., CMAJ, 2009; Takagi et al., Interactive CardioVascular and Thoracic Surgery, 2009).

These findings drove the worldwide marketing suspension in 2007–2008. The subsequent EMA re-review (concluding in February 2012) judged the BART analysis unreliable and supported reinstatement for a restricted population — adults at high risk of major blood loss in isolated CABG — and the Royston 2014 Anaesthesia review summarises the post-reinstatement landscape, including the differing mechanisms and safety profiles of aprotinin and the lysine analogues (Royston, Anaesthesia, 2014).

Regulatory status

  • EU: EMA marketing authorisation reinstated by CHMP recommendation in February 2012 and formally reinstated by the European Commission in 2013, for adults at high risk of major blood loss undergoing isolated CABG. A European post-authorisation safety registry (NAPaR) was established as a condition of reinstatement.
  • US: FDA-approved historically as Trasylol. Bayer suspended US marketing in November 2007 and permanently withdrew the regular marketing authorisation in May 2008 following the BART results. Aprotinin remained accessible in the US only under a restricted, investigational pathway after that point.
  • WADA: Not listed as a prohibited substance — aprotinin is a surgical antifibrinolytic, not a performance-enhancing agent.

Common questions

Is aprotinin still used today? In the European Union and several other jurisdictions, yes — for adults at high bleeding risk undergoing isolated coronary artery bypass surgery, following the EMA's 2012 reinstatement and the European Commission's 2013 decision. In the United States, the regular marketing authorisation has not been restored after the 2008 withdrawal.

Why was aprotinin withdrawn in 2007–2008? Because two observational studies (Mangano 2006 NEJM; Mangano 2007 JAMA) and a randomized trial (BART, Fergusson 2008 NEJM) reported increased mortality and adverse cardiovascular and renal events in aprotinin-treated cardiac surgery patients compared with lysine-analogue antifibrinolytics or no antifibrinolytic.

Why was the European withdrawal reversed? The EMA's 2012 review concluded that the BART trial's mortality result was unreliable on re-analysis, and that the benefit–risk balance favoured aprotinin in a defined high-risk CABG population. The European Commission formally reinstated marketing authorisation in 2013 with a registry requirement (NAPaR) to monitor real-world use.

How is aprotinin different from tranexamic acid? Both reduce surgical bleeding but they intervene at different points in the fibrinolytic cascade. Tranexamic acid (and ε-aminocaproic acid) are lysine analogues that block plasminogen's lysine-binding site so plasmin cannot form efficiently. Aprotinin directly inactivates free plasmin that has already been generated, and also inhibits plasma kallikrein, contributing an anti-inflammatory effect on the cardiopulmonary-bypass response (Royston, Anaesthesia, 2014).

Hypotheses4 directions▾ collapse

Research directions for this peptide, selected from the current sources — hypotheses you can explore and model. None of it is proven yet; tap any one to see the full thinking.

openupdated 2026-06-05

If aprotinin is so tough and compact that it survives almost anything, could scientists attach a tissue-homing signal to it so it only blocks destructive enzymes where they are actually causing disease?

If true, doctors could use aprotinin-like drugs for chronic diseases, not just surgery, because the drug would go exactly where needed instead of affecting the whole body. Patients with inflammatory diseases or slow-healing wounds could benefit without systemic side effects.

The hypothesis
Aprotinin's three-disulfide Kunitz scaffold can tolerate circular permutation or insertion of a targeting peptide without loss of fold integrity, enabling the construction of bifunctional molecules that localize serine-protease inhibition to diseased tissues.
Why it’s plausible
The Kunitz domain is among the most thermodynamically stable small protein folds known, with a melting temperature above 80 degrees C. Its three disulfides create a tightly cross-linked core that tolerates sequence variation in surface loops. Bifunctional fusion proteins and circular permutants of similarly stable scaffolds (e.g., knottins, albumin) have been successfully engineered for targeted delivery. Inserting a tumor-homing or inflammation-targeting peptide into a surface loop of aprotinin, or circularly permuting the termini, could yield a molecule that concentrates protease inhibition at sites of pathological matrix degradation.
Why it matters
This would transform aprotinin from a systemic blunt instrument into a tissue-targeted therapeutic platform, applicable to cancer-associated inflammation, chronic wounds, or fibrotic disease.
Plausibility.65
Novelty.60
Impact.65
Basis · grounding2 computed/notes
[1]
noteAprotinin became one of the most intensively studied small proteins in structural biology and is the archetypal member of the Kunitz protease-inhibitor family, implying that its fold has been exhaustively characterized and is a proven engineering substrate.
[2]
sequenceThe 58-residue sequence contains six cysteines forming three disulfide bonds (Cys5-Cys55, Cys14-Cys38, Cys30-Cys51), creating a highly cross-linked core that should tolerate loop insertions or circular permutation better than single-domain peptides lacking disulfide reinforcement.
openupdated 2026-06-05

If aprotinin works by blocking bleeding enzymes during heart surgery, could it do the same during liver transplants or after major injuries without causing more harm than existing drugs?

If true, surgeons would have a stronger option to control heavy bleeding in operations where current medicines fall short. Patients facing high blood loss could recover faster and need fewer transfusions.

The hypothesis
Aprotinin could reduce bleeding in non-cardiac major surgeries with extracorporeal circulation, such as liver transplantation or trauma resuscitation, if its mechanism of inhibiting contact-phase proteases is broadly transferable beyond the CABG setting.
Why it’s plausible
Aprotinin was reinstated in Europe only for restricted CABG use after a worldwide withdrawal. Its serine-protease inhibition profile (trypsin, kallikrein, plasmin) is not intrinsically heart-specific; contact activation and fibrinolysis occur in any major surgery with large-volume blood loss or circuit exposure. If the safety signal that triggered withdrawal was partly driven by off-label use in higher-risk populations rather than an inherent irreversible toxicity, then narrower patient selection in other surgical arenas might reveal a favorable risk:benefit profile.
Why it matters
Expanding aprotinin beyond CABG could fill an unmet need in surgeries where tranexamic acid is insufficient or contraindicated, without requiring new molecular development.
Plausibility.75
Novelty.35
Impact.60
Basis · grounding1 paper · 1 computed/note
[1]
paper
10.1503/cmaj.081109: meta-analysis of aprotinin vs tranexamic acid showing a summary RR for death of 1.43 (95% CI 0.98-2.08), suggesting the safety concern was a trend rather than definitive harm and that head-to-head comparisons in specific surgical sub-populations remain under-powered.
doi: 10.1503/cmaj.081109
[2]
noteThe drug was withdrawn worldwide in 2007-2008 over safety concerns, then reinstated in the EU in 2012-2013 for a restricted CABG indication after the withdrawal data were re-evaluated, implying that indication-specific risk stratification may alter the benefit:harm balance.
openupdated 2026-06-05

If aprotinin blocks destructive enzymes in the blood, could breathing it in at very low doses protect the lungs during severe pneumonia or ARDS without causing blood clots elsewhere?

If true, hospitals would have a new way to treat life-threatening lung failure using an existing medicine, potentially saving lives during flu seasons or pandemics when ventilators are not enough.

The hypothesis
Low-dose aprotinin, at concentrations below full systemic antifibrinolysis, could limit alveolar-capillary leak in acute respiratory distress syndrome (ARDS) by local inhibition of neutrophil elastase and plasmin-mediated matrix degradation.
Why it’s plausible
Neutrophil elastase and plasmin are both serine proteases implicated in ARDS pathophysiology: they degrade basement membrane proteins, activate matrix metalloproteinases, and amplify inflammation. Aprotinin inhibits both enzymes. If inhaled or delivered at sub-anticoagulant doses, it might attenuate lung injury without the systemic thrombotic risks that complicated its high-dose cardiac-surgery use. The Kunitz fold is highly stable and resists proteolysis, making it suitable for aerosol delivery.
Why it matters
ARDS lacks effective pharmacotherapy; repurposing an existing, well-characterized molecule with a known safety database could accelerate clinical translation.
Plausibility.50
Novelty.55
Impact.70
Basis · grounding1 paper · 1 computed/note
[1]
noteAprotinin is a 58-amino-acid serine protease inhibitor that blocks bleeding-promoting enzymes, and its Kunitz-type fold is stabilised by three disulfide bonds, giving it exceptional proteolytic stability that would survive pulmonary delivery.
[2]
paper
10.1503/cmaj.081109: the safety signal for aprotinin was dose- and indication-dependent (restricted reinstatement in CABG), suggesting that lower or localized dosing in a different organ context might separate efficacy from systemic harm.
doi: 10.1503/cmaj.081109
openupdated 2026-06-05

If the tail end of aprotinin touches different bleeding enzymes in different ways, could changing that tail make the drug stop bleeding without the side effects that got it banned?

If true, doctors could use a safer, customized version of an existing drug instead of inventing an entirely new one. Patients would face lower risks of kidney damage or dangerous clots after surgery.

The hypothesis
The C-terminal region of aprotinin (residues 46-58, containing the RNNFKSAEDCMRTCGGA tail) contributes disproportionately to its inhibition of plasmin relative to trypsin, and swapping or truncating this segment could yield a variant with altered fibrinolysis:coagulation selectivity.
Why it’s plausible
Canonical BPTI/aprotinin structure-function studies mapped the reactive-center loop (around Lys15) as the primary trypsin/kallikrein engagement site, but the C-terminal tail extends away from the core Kunitz domain and makes additional contacts with larger protease exosites. In plasmin, the extended substrate-binding cleft and kringle domains create a larger interaction surface than trypsin's shallow active-site groove. The sequence contains a Cys55 that forms the Cys5-Cys55 disulfide, but residues 50-58 remain flexible; altering them may therefore modulate plasmin inhibition without destroying the core fold.
Why it matters
A plasmin-selective aprotinin variant could preserve antifibrinolytic efficacy while reducing contact-phase suppression that has been linked to renal and thrombotic adverse effects.
Plausibility.45
Novelty.70
Impact.55
Basis · grounding2 computed/notes
[1]
sequenceThe 58-residue sequence RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGA contains Cys at positions 5, 14, 30, 38, 51, and 55, consistent with the three canonical disulfides; residues 46-58 include the unique C-terminal extension beyond the minimal Kunitz domain.
[2]
noteAprotinin is described as a Kunitz-type fold stabilised by three disulfide bonds, with the bridges essential to its compact pear-shaped structure and ability to engage active-site serines, implying that regions outside the disulfide core are available for engineering without destabilising the fold.
details expand to inspect
3-letter notation
Arg-Pro-Asp-Phe-Cys-Leu-Glu-Pro-Pro-Tyr-Thr-Gly-Pro-Cys-Lys-Ala-Arg-Ile-Ile-Arg-Tyr-Phe-Tyr-Asn-Ala-Lys-Ala-Gly-Leu-Cys-Gln-Thr-Phe-Val-Tyr-Gly-Gly-Cys-Arg-Ala-Lys-Arg-Asn-Asn-Phe-Lys-Ser-Ala-Glu-Asp-Cys-Met-Arg-Thr-Cys-Gly-Gly-Ala
citationbibtex
peptidemodel (2026). Aprotinin (Trasylol): cuts bleeding during heart surgery (pep-04443, v1). PeptideModel. https://peptidemodel.com/card/pep-04443
@peptide{pep04443,
  sequence = {RPDFCLEPPYTGPCKARIIRYFYNAKAGLCQTFVYGGCRAKRNNFKSAEDCMRTCGGA},
  target   = {},
  author   = {peptidemodel},
  year     = {2026},
  status   = {bioassayed}
}
clinical trials 61 on ct.gov · 16 on EUCTR · checked 2026-05-22
ct.gov trials 61
with results 10
EUCTR 16
PubMed RCT 346
by phase
2phase 48no phase
by status
4completed1recruiting1not yet recruiting4unknown
references 23 papers
[2]
The Risk Associated with Aprotinin in Cardiac Surgery
Mangano, D. et al. New England Journal of Medicine 2006
supporting
[7] supporting
[8]
Aprotinin in der Kardiochirurgie
Bremerich, D. et al. Der Anaesthesist 2006
supporting
[9]
Judging the Safety of Aprotinin
New England Journal of Medicine 2006
supporting
[11] supporting
[12]
Bleeding in cardiac surgery: The use of aprotinin does not affect survival
Pagano, D. et al. The Journal of Thoracic and Cardiovascular Surgery 2008
supporting
[13] supporting
[14]
The Aprotinin Story — Is BART the Final Chapter?
Ray, W. et al. New England Journal of Medicine 2008
supporting
[16]
Aprotinin: 1 year on
Dietrich, W. Current Opinion in Anaesthesiology 2009
supporting
[18]
Aprotinin revisited
DeAnda, A. et al. The Journal of Thoracic and Cardiovascular Surgery 2012
supporting
[19] supporting
[21]
Aprotinin and bleeding in profoundly hypothermic perfusion
WESTABY, S. et al. European Journal of Cardio-Thoracic Surgery 1994
supporting
[23]
Effect of aprotinin on vascular reactivity of coronary bypass grafts
Allen, S. et al. The Journal of Thoracic and Cardiovascular Surgery 1997
supporting
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