The influence of various structural parameters of semisynthetic sulfated polysaccharides on the P-selectin inhibitory capacity.
Selectin-mediated leukocyte rolling along the endothelium is of key importance for maintaining the cellular immune response. The anti-inflammatory activities of heparin have partly been related to inhibition of P-selectin binding. Heparin, however, suffers from its heterogeneous variable structure, the animal origin and multiple in vivo effects. As P-selectin is a promising target for anti-inflammatory approaches, we focused on P-selectin inhibition by other sulfated polysaccharides and compared them with six heparins. We examined 15 structurally defined semisynthetic sulfated glucans, non-animal-derived from the linear glucans phycarin, curdlan or pullulan. The derivatives gradually differ in their degree of sulfation, molecular weight, and glycosidic linkage. The inhibitory capacity was analysed in a parallel plate flow chamber, detecting the rolling of U937 cells on P-selectin layers. Unfractionated heparins displayed variabilities between different preparations. Considering fractionated heparins, exceeding of a minimal mass is essential for activity. Comparing the glucan sulfates, charge density is the most important parameter for P-selectin binding. Highly sulfated derivatives are excellent inhibitors, the reduced cell binding up to 16.2+/-6.4% strongly exceeded the heparin activities. Molecular weight is of minor effects, while glycosidic backbone linkage holds certain importance. To check the P-selectin inhibition in vivo, heparin and one phycarin sulfate were tested using intravital microscopy of microvasculature in mice. Both compounds significantly reduced the rolling fractions of activated platelets on endothelium as effective as a blocking P-selectin antibody. Our study indicates that semisynthetic glucan sulfates with optimal structures block P-selectin excellently and might become promising candidates for anti-inflammatory drugs to replace heparin for certain applications.
Fritzsche
Juliane
J
Institute of Pharmaceutical Chemistry, Rheinische Friedrich Wilhelms University Bonn, Germany.
Alban
Susanne
S
Ludwig
Ralf J
RJ
Rubant
Simone
S
Boehncke
Wolf-Henning
WH
Schumacher
Gabriele
G
Bendas
Gerd
G
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000925:Anticoagulants / Q000737:chemistry / Q000494:pharmacology; D001792:Blood Platelets / Q000502:physiology; D002448:Cell Adhesion / Q000187:drug effects; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D005936:Glucans / Q000737:chemistry / Q000494:pharmacology; D006493:Heparin / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D036904:Leukocyte Rolling / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D019007:P-Selectin / Q000276:immunology / Q000494:pharmacology* / Q000502:physiology; D015539:Platelet Activation / Q000187:drug effects; D011134:Polysaccharides / Q000737:chemistry / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013431:Sulfates / Q000737:chemistry; D020298:U937 Cells; D047071:beta-Glucans / Q000737:chemistry / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000925:Anticoagulants; D005936:Glucans; D019007:P-Selectin; D011134:Polysaccharides; D013431:Sulfates; D047071:beta-Glucans; C078962:glucan sulfate; C009109:pullulan; D006493:Heparin; C033363:beta-1,3-glucan; C063130:curdlan sulfate