Anti-inflammatory activity of c(ILDV-NH(CH2)5CO), a novel, selective, cyclic peptide inhibitor of VLA-4-mediated cell adhesion.
1. Small, N- to C-terminal cyclized peptides containing the leucyl-aspartyl-valine (LDV) motif from fibronectin connecting segment-1 (CS-1) have been investigated for their effects on the adhesion of human T-lymphoblastic leukaemia cells (MOLT-4) to human plasma fibronectin in vitro mediated by the integrin Very Late Antigen (VLA)-4 (alpha4beta1, CD49d/CD29). 2. Cyclo(-isoleucyl-leucyl-aspartyl-valyl-aminohexanoyl-) (c(ILDV-NH(CH2)5CO)) was approximately 5 fold more potent (IC50 3.6+/-0.44 microM) than the 25-amino acid linear CS-1 peptide. Cyclic peptides containing two more or one less methylene groups had similar potency to c(ILDV-NH(CH2)5CO) while a compound containing three less methylene groups, c(ILDV-NH(CH2)2CO), was inactive at 100 microM. 3. c(ILDV-NH(CH2)5CO) had little effect on cell adhesion mediated by two other integrins, VLA-5 (alpha5,beta1, CD49e/CD29) (K562 cell adhesion to fibronectin) or Leukocyte Function Associated molecule-1 (LFA-1, alphabeta2, CD11a/CD18) (U937 cell adhesion to Chinese hamster ovary cells transfected with intercellular adhesion molecule-1) at concentrations up to 300 microM. 4. c(ILDV-NH(CH2)5CO) inhibited ovalbumin delayed-type hypersensitivity or oxazolone contact hypersensitivity in Balb/c mice when dosed continuously from subcutaneous osmotic mini-pumps (0.1-10 mg kg(-1) day(-1)). Maximum inhibition (approximately 40%) was similar to that caused by the monoclonal antibody PS/2 (7.5 mg kg(-1) i.v.) directed against the alpha4 integrin subunit. 5. c(ILDV-NH(CH2)5CO) also inhibited oxazolone contact hypersensitivity when dosed intravenously 20 h after oxazolone challenge (1-10 mg kg(-1)). Ear swelling was reduced at 3 h and 4 h but not at 1 h and 2 h post-dose (10 mg kg(-1)). 6. Small molecule VLA-4 inhibitors derived from c(ILDV-NH(CH2)5CO) may be useful as anti-inflammatory agents.
Haworth
D
D
CM&M Research Department, Zeneca Pharmaceuticals, Macclesfield, Cheshire.
Rees
A
A
Alcock
P J
PJ
Wood
L J
LJ
Dutta
A S
AS
Gormley
J J
JJ
Jones
H B
HB
Jamieson
A
A
Reilly
C F
CF
D000595:Amino Acid Sequence; D000818:Animals; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000494:pharmacology*; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D003877:Dermatitis, Contact / Q000188:drug therapy / Q000276:immunology; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D006968:Hypersensitivity, Delayed / Q000188:drug therapy / Q000276:immunology; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology; D039041:Integrin alpha4beta1; D016023:Integrins / Q000037:antagonists & inhibitors* / Q000502:physiology; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D036341:Intercellular Signaling Peptides and Proteins; D004915:Leukemia, Erythroblastic, Acute / Q000473:pathology; D015458:Leukemia, T-Cell / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000276:immunology; D010081:Oxazolone / Q000276:immunology; D010455:Peptides / Q000494:pharmacology*; D051381:Rats; D016201:Receptors, Lymphocyte Homing / Q000037:antagonists & inhibitors* / Q000502:physiology; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D014162:Transfection
D000894:Anti-Inflammatory Agents, Non-Steroidal; D005353:Fibronectins; D039041:Integrin alpha4beta1; D016023:Integrins; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; D016201:Receptors, Lymphocyte Homing; C076687:CS1 peptide; D018799:Intercellular Adhesion Molecule-1; D010081:Oxazolone; D010047:Ovalbumin