The use of the Chandler loop to examine the interaction potential of NXY-059 on the thrombolytic properties of rtPA on human thrombi in vitro

N. J. Mutch, Norma Ross Moore, C. Mattsson, H. Jonasson, A. R. Green, Nuala Ann Booth

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background and purpose: Recombinant tissue-type plasminogen activator (rtPA) is the only globally approved treatment for acute ischaemic stroke. Other potential treatments might be administered with rtPA, making it important to discover whether compounds interfere with rtPA-induced lysis. We evaluated methods for examining the effect of the neuroprotectant NXY-059 on the lytic property of rtPA.
Experimental approach: Plasma clot formation and lysis in the presence of rtPA and NXY-059 was measured as the change in plasma turbidity. The effect of NXY-059 on rtPA-induced lysis was similarly assessed on preformed clots. Lysis of the thrombus formed in a Chandler loop measured release of fluorescent-tagged fibrinogen that had been incorporated during thrombus formation. Thrombi were exposed to both rtPA and NXY-059 throughout lysis in the presence of 80% autologous plasma and the release of label during lysis was measured.
Key results: Data interpretation is limited in the clot lysis experiments because either the rtPA was present during clot formation or the drug was added to a clot formed in static conditions. In contrast, thrombi were formed in dynamic flow conditions in the Chandler loop and the time course of lysis in plasma was examined. rtPA increased thrombolysis and the antifibrinolytic trans-4-(aminomethyl) cyclohexane carboxylic acid (AMCA) inhibited lysis. Lysis induced by rtPA was unaltered by NXY-059.
Conclusions and implications: The Chandler loop method provides a reliable technique for examining the effect of compounds on rtPA-induced lysis in vitro and demonstrated that NXY-059 does not alter rtPA-induced lysis at clinically relevant concentrations of either drug.

Original languageEnglish
Pages (from-to)124-131
Number of pages8
JournalBritish Journal of Pharmacology
Volume153
Issue number1
DOIs
Publication statusPublished - Jan 2008

Keywords

  • recombinant tissue-type plasminogen activator
  • NXY-059
  • rtPA
  • thrombolysis
  • Chandler loop
  • acute ischemic stoke
  • acute myocardial infarction
  • tissue plasmogen activator
  • radical trapping ability
  • cerebral ischemia
  • clot lysis
  • S-PBN
  • alteplase
  • blood
  • pharmacokinetics

Cite this

The use of the Chandler loop to examine the interaction potential of NXY-059 on the thrombolytic properties of rtPA on human thrombi in vitro. / Mutch, N. J.; Moore, Norma Ross; Mattsson, C.; Jonasson, H.; Green, A. R.; Booth, Nuala Ann.

In: British Journal of Pharmacology, Vol. 153, No. 1, 01.2008, p. 124-131.

Research output: Contribution to journalArticle

@article{d140b0ab230c4497a8c1b922365dc83f,
title = "The use of the Chandler loop to examine the interaction potential of NXY-059 on the thrombolytic properties of rtPA on human thrombi in vitro",
abstract = "Background and purpose: Recombinant tissue-type plasminogen activator (rtPA) is the only globally approved treatment for acute ischaemic stroke. Other potential treatments might be administered with rtPA, making it important to discover whether compounds interfere with rtPA-induced lysis. We evaluated methods for examining the effect of the neuroprotectant NXY-059 on the lytic property of rtPA. Experimental approach: Plasma clot formation and lysis in the presence of rtPA and NXY-059 was measured as the change in plasma turbidity. The effect of NXY-059 on rtPA-induced lysis was similarly assessed on preformed clots. Lysis of the thrombus formed in a Chandler loop measured release of fluorescent-tagged fibrinogen that had been incorporated during thrombus formation. Thrombi were exposed to both rtPA and NXY-059 throughout lysis in the presence of 80{\%} autologous plasma and the release of label during lysis was measured. Key results: Data interpretation is limited in the clot lysis experiments because either the rtPA was present during clot formation or the drug was added to a clot formed in static conditions. In contrast, thrombi were formed in dynamic flow conditions in the Chandler loop and the time course of lysis in plasma was examined. rtPA increased thrombolysis and the antifibrinolytic trans-4-(aminomethyl) cyclohexane carboxylic acid (AMCA) inhibited lysis. Lysis induced by rtPA was unaltered by NXY-059. Conclusions and implications: The Chandler loop method provides a reliable technique for examining the effect of compounds on rtPA-induced lysis in vitro and demonstrated that NXY-059 does not alter rtPA-induced lysis at clinically relevant concentrations of either drug.",
keywords = "recombinant tissue-type plasminogen activator, NXY-059, rtPA, thrombolysis, Chandler loop, acute ischemic stoke, acute myocardial infarction, tissue plasmogen activator, radical trapping ability, cerebral ischemia, clot lysis, S-PBN, alteplase, blood, pharmacokinetics",
author = "Mutch, {N. J.} and Moore, {Norma Ross} and C. Mattsson and H. Jonasson and Green, {A. R.} and Booth, {Nuala Ann}",
year = "2008",
month = "1",
doi = "10.1038/sj.bjp.0707543",
language = "English",
volume = "153",
pages = "124--131",
journal = "British Journal of Pharmacology",
issn = "0007-1188",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - The use of the Chandler loop to examine the interaction potential of NXY-059 on the thrombolytic properties of rtPA on human thrombi in vitro

AU - Mutch, N. J.

AU - Moore, Norma Ross

AU - Mattsson, C.

AU - Jonasson, H.

AU - Green, A. R.

AU - Booth, Nuala Ann

PY - 2008/1

Y1 - 2008/1

N2 - Background and purpose: Recombinant tissue-type plasminogen activator (rtPA) is the only globally approved treatment for acute ischaemic stroke. Other potential treatments might be administered with rtPA, making it important to discover whether compounds interfere with rtPA-induced lysis. We evaluated methods for examining the effect of the neuroprotectant NXY-059 on the lytic property of rtPA. Experimental approach: Plasma clot formation and lysis in the presence of rtPA and NXY-059 was measured as the change in plasma turbidity. The effect of NXY-059 on rtPA-induced lysis was similarly assessed on preformed clots. Lysis of the thrombus formed in a Chandler loop measured release of fluorescent-tagged fibrinogen that had been incorporated during thrombus formation. Thrombi were exposed to both rtPA and NXY-059 throughout lysis in the presence of 80% autologous plasma and the release of label during lysis was measured. Key results: Data interpretation is limited in the clot lysis experiments because either the rtPA was present during clot formation or the drug was added to a clot formed in static conditions. In contrast, thrombi were formed in dynamic flow conditions in the Chandler loop and the time course of lysis in plasma was examined. rtPA increased thrombolysis and the antifibrinolytic trans-4-(aminomethyl) cyclohexane carboxylic acid (AMCA) inhibited lysis. Lysis induced by rtPA was unaltered by NXY-059. Conclusions and implications: The Chandler loop method provides a reliable technique for examining the effect of compounds on rtPA-induced lysis in vitro and demonstrated that NXY-059 does not alter rtPA-induced lysis at clinically relevant concentrations of either drug.

AB - Background and purpose: Recombinant tissue-type plasminogen activator (rtPA) is the only globally approved treatment for acute ischaemic stroke. Other potential treatments might be administered with rtPA, making it important to discover whether compounds interfere with rtPA-induced lysis. We evaluated methods for examining the effect of the neuroprotectant NXY-059 on the lytic property of rtPA. Experimental approach: Plasma clot formation and lysis in the presence of rtPA and NXY-059 was measured as the change in plasma turbidity. The effect of NXY-059 on rtPA-induced lysis was similarly assessed on preformed clots. Lysis of the thrombus formed in a Chandler loop measured release of fluorescent-tagged fibrinogen that had been incorporated during thrombus formation. Thrombi were exposed to both rtPA and NXY-059 throughout lysis in the presence of 80% autologous plasma and the release of label during lysis was measured. Key results: Data interpretation is limited in the clot lysis experiments because either the rtPA was present during clot formation or the drug was added to a clot formed in static conditions. In contrast, thrombi were formed in dynamic flow conditions in the Chandler loop and the time course of lysis in plasma was examined. rtPA increased thrombolysis and the antifibrinolytic trans-4-(aminomethyl) cyclohexane carboxylic acid (AMCA) inhibited lysis. Lysis induced by rtPA was unaltered by NXY-059. Conclusions and implications: The Chandler loop method provides a reliable technique for examining the effect of compounds on rtPA-induced lysis in vitro and demonstrated that NXY-059 does not alter rtPA-induced lysis at clinically relevant concentrations of either drug.

KW - recombinant tissue-type plasminogen activator

KW - NXY-059

KW - rtPA

KW - thrombolysis

KW - Chandler loop

KW - acute ischemic stoke

KW - acute myocardial infarction

KW - tissue plasmogen activator

KW - radical trapping ability

KW - cerebral ischemia

KW - clot lysis

KW - S-PBN

KW - alteplase

KW - blood

KW - pharmacokinetics

U2 - 10.1038/sj.bjp.0707543

DO - 10.1038/sj.bjp.0707543

M3 - Article

C2 - 17982476

VL - 153

SP - 124

EP - 131

JO - British Journal of Pharmacology

JF - British Journal of Pharmacology

SN - 0007-1188

IS - 1

ER -