Long duration of anticoagulant activity and protective effects of acharan sulfate in vivo.
BACKGROUND
We previously reported that a new glycosaminoglycan, acharan sulfate (AS) from the African giant snail Achatina fulica showed anticoagulant activity in vitro, but was much less active when compared to heparin. In the present study, the anticoagulant activity of AS was investigated in vivo.


METHODS
AS and heparin were administered to mice and rats in various doses and the anticoagulant activities were measured by aPTT assay. Both were also compared in a thrombin-induced lethality animal model. As one of the possible mechanisms, AS-thrombin interaction was studied by using surface plasmon resonance spectroscopy.


RESULTS
Intravenous administration of AS to mice prolonged the clotting time (aPTT) in a time and dose-dependent manner. Although the anticoagulant activity was low in rats, it steadily increased over 5 h after administration of AS (30 mg/kg). In contrast, the increase in aPTT induced by 5 mg/kg of heparin was restored to a normal level after 3 h. In a thrombin-induced lethality model in mice, AS (20 mg/kg) protected against lethality by 80%, while heparin (20 mg/kg) did not show any protective activity beyond 3.5 h post-administration. AS could be also detected in plasma even 5 h after i.v. administration to rats. The binding constant (K(D)) of AS to thrombin was 7.27 x 10(-6) M, corresponding to moderate binding affinity.


CONCLUSIONS
These results show that the longer duration of AS in blood could prolong the clotting time determined by aPTT and offering protection against thrombin-induced lethality. One possible mechanism may result from AS-thrombin interaction.
Li
Da-Wei
DW
Natural Products Research Institute, College of Pharmacy, Seoul National University, 28 Yeonkun-Dong, Jongno-Ku, Seoul 110-460, South Korea.
Lee
In Sun
IS
Sim
Joon-Soo
JS
Toida
Toshihiko
T
Linhardt
Robert J
RJ
Kim
Yeong Shik
YS
D000818:Animals; D000925:Anticoagulants / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D001777:Blood Coagulation / Q000187:drug effects; D001780:Blood Coagulation Tests; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D006025:Glycosaminoglycans / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use; D006493:Heparin / Q000008:administration & dosage; D007275:Injections, Intravenous; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D051381:Rats; D017207:Rats, Sprague-Dawley; D012908:Snails / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D013997:Time Factors
D000925:Anticoagulants; D006025:Glycosaminoglycans; C106771:acharan sulfate; D006493:Heparin; D013917:Thrombin