title: Non-Anticoagulant Fractions of Enoxaparin Suppress Inflammatory Cytokine Release from Peripheral Blood Mononuclear Cells of Allergic Asthmatic Individuals.
abstract: BACKGROUND
  Enoxaparin, a low-molecular-weight heparin, is known to possess anti-inflammatory properties. However, its clinical exploitation as an anti-inflammatory agent is hampered by its anticoagulant effect and the associated risk of bleeding.
  
  
  OBJECTIVE
  The aim of the current study was to examine the ability of non-anticoagulant fractions of enoxaparin to inhibit the release of key inflammatory cytokines in primed peripheral blood mononuclear cells derived from allergic mild asthmatics.
  
  
  METHODS
  Peripheral blood mononuclear cells from allergic asthmatics were activated with phytohaemag glutinin (PHA), concanavalin-A (ConA) or phorbol 12-myristate 13-acetate (PMA) in the presence or absence of enoxaparin fractions before cytokine levels were quantified using specific cytokine bead arrays. Together with nuclear magnetic resonance analysis,time-dependent and target-specific effects of enoxaparin fractions were used to elucidate structural determinants for their anti-inflammatory effect and gain mechanistic insights into their anti-inflammatory activity.
  
  
  RESULTS
  Two non-anticoagulant fractions of enoxaparin were identified that significantly inhibited T-cell activation. A disaccharide fraction of enoxaparin inhibited the release of IL-4, IL-5, IL-13 and TNF-α by more than 57% while a tetrasaccharide fraction was found to inhibit the release of tested cytokines by more than 68%. Our data suggest that the observed response is likely to be due to an interaction of 6-O-sulfated tetrasaccharide with cellular receptor(s).
  
  
  CONCLUSIONS
  The two identified anti-inflammatory fractions lacked anticoagulant activity and are therefore not associated with risk of bleeding. The findings highlight the potential therapeutic use of enoxaparin-derived fractions, in particular tetrasaccharide, in patients with chronic inflammatory disorders.
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Madhur D
authors.initials: MD
authors.lastname: Shastri
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Niall
authors.initials: N
authors.lastname: Stewart
authors.affiliation: Central Science Laboratory, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: James
authors.initials: J
authors.lastname: Horne
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Syed Tabish R
authors.initials: ST
authors.lastname: Zaidi
authors.affiliation: Breathe Well Centre of Research Excellence for Chronic Respiratory Disease and Lung Ageing, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Sukhwinder Singh
authors.initials: SS
authors.lastname: Sohal
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia; Breathe Well Centre of Research Excellence for Chronic Respiratory Disease and Lung Ageing, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Peterson
authors.affiliation: Menzies Institute for Medical Research, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Heinrich
authors.initials: H
authors.lastname: Korner
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Nuri
authors.initials: N
authors.lastname: Gueven
authors.affiliation: Pharmacy, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
authors.forename: Rahul P
authors.initials: RP
authors.lastname: Patel
mesh_terms: D000328:Adult; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000302:isolation & purification / Q000494:pharmacology*; D001249:Asthma / Q000276:immunology / Q000473:pathology; D005591:Chemical Fractionation; D003208:Concanavalin A / Q000494:pharmacology; D017984:Enoxaparin / Q000737:chemistry*; D005260:Female; D006801:Humans; D018793:Interleukin-13 / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D015847:Interleukin-4 / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D015848:Interleukin-5 / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000187:drug effects* / Q000276:immunology / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009844:Oligosaccharides / Q000302:isolation & purification / Q000494:pharmacology*; D010835:Phytohemagglutinins / Q000494:pharmacology; D061251:Primary Cell Culture; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000096:biosynthesis
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D017984:Enoxaparin; C578095:IL13 protein, human; C508598:IL4 protein, human; C508599:IL5 protein, human; D018793:Interleukin-13; D015848:Interleukin-5; D009844:Oligosaccharides; D010835:Phytohemagglutinins; D014409:Tumor Necrosis Factor-alpha; D003208:Concanavalin A; D015847:Interleukin-4; D013755:Tetradecanoylphorbol Acetate