• Journal of Pharmaceutical Investigation
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• 제39권5호
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• pp.373-379
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• 2009

This study was conducted to develop a pharmacokinetic-pharmacodynamic (PK/PD) model of a direct thrombin inhibitor, argatroban to predict the concentration-effect profiles in rats. Argatroban was i.v. injected to rats at 0. 2, 0.8 and 3.2 mg/kg doses (n = 4-5 per dose), and plasma drug levels were determined by a validated LC/MS/MS assay. The pharmacokinetics of argatroban was linear over the i.v. dose range studied. The thrombin time (TT) and the activated partial thromboplastin time (aPTT) were measured in rat plasma and they were found to linearly increase with increasing the dose. A 2-compartment pharmacokinetic model linked with an indirect response pharmacodynamic model was successfully utilized to evaluate the drug concentration-response relationship.

• 생명과학회지
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• 제20권6호
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• pp.922-928
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• 2010

한방 생약재로부터 안전하면서도 신규의 항혈전제를 개발하기 위해, 현재 국내에서 유통되고 있는 국내 및 국외산 한방 생약재 33종을 대상으로 40종의 에탄올 추출물을 조제하고, 이들의 thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (aPTT)을 평가하였다. 생약재중 중국산은 28종으로 대부분이 국외산이었으며, 부위별로는 종자 및 뿌리 부위가 19종을 차지하였다. 평균 수분함량은 $6.85{\pm}2.26%$, 평균 추출율은 $5.27{\pm}4.25%$를 나타내었다. 조제된 시료의 TT평가 결과, 사인(중국), 고본, 팔각향, 계지, 당삼, 계혈등, 행인, 오배자, 괄루근 및 포황에서 우수한 트롬빈 저해활성을 확인하였으며, 다양한 농도에서 평가 결과 계혈등, 행인, 팔각향, 계지, 오배자 추출물 순으로 트롬빈 저해능이 강력함을 확인하였다. PT 평가 결과 오배자, 계혈등, 행인에서, aPTT 평가결과 오배자 및 계혈등에서 우수한 항혈전능을 확인하였다. 한편 추출물의 인간적혈구 용혈활성을 평가한 결과, 계혈등, 팔각향, 계지, 오배자 추출물은 $500\;{\mu}g/ml$ 농도에서도 용혈활성이 없음을 확인하여 이들 4종을 최종 선정하였다. 본 연구결과는 이미 대량생산이 확립된 한방생약제로부터 우수한 항혈전제 개발이 가능함을 제시하고 있으며, 트롬빈 및 혈장 응고 인자에 대한 특이 저해제 개발이 가능함을 제시하고 있다.

• 생명과학회지
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• 제20권11호
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• pp.1640-1647
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• 2010

혈전성 질환을 예방, 개선할 수 있는 안전한 항혈전제를 개발하기 위해, 일반적으로 항혈전 효과가 있다고 알려진 식용 해조류 35종(갈조류 17종, 홍조류 11종 및 녹조류 7종)을 대상으로 메탄올 추출물을 조제하고, 이들의 thrombin time (TT), prothrombin time (PT), activated partial thromboplastin time (aPTT)을 평가하였다. 트롬빈 저해(TT)의 경우 35종 중에서 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 야마다모자반에서 강력한 활성을, 프로트롬빈 저해(PT)의 경우 트롬빈 저해활성이 나타난 7종 중 야마다모자반을 제외한 6종에서 강력한 활성을 확인하였으며, 혈액응고인자 저해(aPTT)에서는 감태, 곰피, 대황, 넓패, 패, 알송이모자반 및 톳에서 강력한 활성이 나타났다. 선별된 8종의 시료를 대상으로 다양한 농도에서 항혈전 활성을 평가한 결과 알송이모자반에서 가장 강력한 활성을 확인하였으며, 곰피, 대황 및 넓패가 효과적이었다. 항혈전 활성과 해조류 추출물의 총 flavonoid, 총 polyphenol, 총당 및 환원당 함량과의 상관 관계를 검토한 결과, 활성물질은 flavonoid성 물질로 추측되었으며, 선별된 8종은 총 flavonoid 함량 및 총 polyphenol 함량이 매우 높은 특징을 나타내었다. 본 연구 결과는 경제적이면서 대량공급이 가능한 식용 해조류로부터 신규의 안전한 항혈전제 개발이 가능함을 제시하며, 야마다모자반과 톳은 트롬빈 또는 혈장내 응고 인자에 대한 특이 저해제로 개발 가능함을 제시하고 있다.

• BMB Reports
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• 제45권4호
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• pp.221-226
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• 2012

Curcumin, a polyphenol responsible for the yellow color of the curry spice turmeric, possesses antiinflammatory, antiproliferative and antiangiogenic activities. However, anticoagulant activities of curcumin have not been studied. Here, the anticoagulant properties of curcumin and its derivative (bisdemethoxycurcumin, BDMC) were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT) as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed that curcumin and BDMC prolonged aPTT and PT significantly and inhibited thrombin and FXa activities. They inhibited the generation of thrombin or FXa. In accordance with these anticoagulant activities, curcumin and BDMC showed anticoagulant effect in vivo. Surprisingly, these anticoagulant effects of curcumin were better than those of BDMC indicating that methoxy group in curcumin positively regulated anticoagulant function of curcumin. Therefore, these results suggest that curcumin and BDMC possess antithrombotic activities and daily consumption of the curry spice turmeric might help maintain anticoagulant status.

• BMB Reports
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• 제47권7호
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• pp.376-381
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• 2014

Enzymatic oxidation of pyrogallol was efficiently transformed to an oxidative product, purpurogallin (PPG). Here, the anticoagulant activities of PPG were examined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), and the activities of thrombin and activated factor X (FXa). And, the effects of PPG on expression of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were evaluated in tumor necrosis factor (TNF)-${\alpha}$ activated human umbilical vein endothelial cells (HUVECs). Treatment with PPG resulted in prolonged aPTT and PT and inhibition of the activities of thrombin and FXa, as well as inhibited production of thrombin and FXa in HUVECs. In addition, PPG inhibited thrombin-catalyzed fibrin polymerization and platelet aggregation. PPG also elicited anticoagulant effects in mice. In addition, treatment with PPG resulted in significant reduction of the PAI-1 to t-PA ratio. Collectively, PPG possesses antithrombotic activities and offers a basis for development of a novel anticoagulant.

• 생명과학회지
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• 제17권4호
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• pp.535-539
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• 2007

식용 및 약용으로 광범위하게 이용되고 있는 산사자의 유용 생리활성 검토를 목적으로 산사자 추출물 및 분획물을 조제하여, 혈액내의 혈전 생성에 직접 관여하는 인간 트롬빈의 thrombin time(TT) 및 activated partial thromboplastin time(aPTT) 증가 활성을 측정하여 항혈전 활성을 평가하였다. 산사자의 항혈전 물질은 물 추출보다는 메탄을 추출이 적합하며(추출효율 40.4%), butanol 분획에서 가장 강력한 활성을 나타내었다. 산사자의 butanol 분획물은 1.25 mg/ml의 농도에서 835%의 TT 증가, 315%의 aPTT 증가를 보여 아스피린보다 2.9배 이상의 강력한 항혈전 활성을 나타내었다. 이러한 항혈전 활성은 기존의 보고된 quercetin, kaempferol, myricetin 및 rutin과는 무관하였다. 또한 산사자의 활성물질은 0.5 N HCl 처리시 120분 이상 안정하였으나, $100^{\circ}C$, 30분 열처리에는 85% 이상의 활성소실이 나타나 열에 매우 민감한 물질로 확인되었다. 본 연구결과는, 비열 처리와 적합한 가공 공정이 가능하다면 산사자 추출물이 혈액순환 장해 및 혈관계 질환 예방 및 치료제로 개발 가능함을 제시하고 있다.

• 대한본초학회지
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• 제26권4호
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• pp.125-132
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• 2011

Objectives : An aim of study is to investigate effects of curculiginis rhizoma in vitro (factor Xa (FXa) inhibitor assay, prothrombinase assay, prothrombin time (PT) assay, activated partial thromboplastin time (aPTT) assay) and in vivo experiment (blood clotting time, thromboxane B2 content assay in serum and weight of thrombus by AV-shunt rat model). Methods : We gained a human serum and used serum in vitro study such as factor X activity (FXa) inhibition, prothrombinase inhibition, prothrombin time (PT) and activated partial thromboplastin time. Fifteen SD rats were divided into three groups (intact control group and two experimental group treated with extract of Curculiginis Rhizoma(ECR)). Rats were orally administrated DW (intact control group), 600 mg/kg concertration of ECR and 200 mg/kg concertration of ECR. After one hour, we anesthetized rats and made arteriovenous (AV) shunt rat models to study weights of thrombus, took a hole blood to study content of thromboxane B2 and blood clotting time. Results : In vitro, ECR increased a inhibitory activity of FXa, prothrombinase and aPTT compared than intact control group. Especially ECR made significant increase of FXa and prothrombinase inhibitory activity (p<0.05, p<0.01). And PT were increased in ECR control group compared with intact control group. In vivo, a blood clotting time of experiment group treated with ECR 600 mg/kg were significantly increased compared with that of intact control group (p<0.05) and content of thromboxane B2 was significantly decreased in group treated with ECR 600 mg/kg in seum. The weight of thrombus were significantly reduced in group treated with ECR 600 mg/kg compared with intact control group (p<0.05). But in vivo experiment study, those of group treated with ECR 200 mg/kg were reduced compared with those of intact control group without statistical significance. Conclusions : ECR has a antithromboic activity in internal course with inhibitory activity of FXa and prothrombinase in vitro, it required to research more study for effective compounds.

• 생약학회지
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• 제35권1호통권136호
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• pp.52-61
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• 2004

Inhibitory activities of 264 methanol extracts, which were prepared from different parts of 210 kinds of wild and medicinal plants, against human thrombin were evaluated. Based on the anti-coagulation activity determined by thrombin time and activated partial thromboplastin time, the 14 extracts were screened. The fibrinolytic activity, heat stability and inhibition of other proteolytic digestive enzymes, such as pepsin, papain, trypsin and chymotrypsin, of the 14 extracts were further determined, and Ginko biloba (herba), Ephedra sinica (radix), Reynoutria elliptica (herba), Amomum tsao-ko Crevost (fructus), and Magnolia officinalis Rehd. et Wils (bark) were finally selected as possible plant sources for anti-thrombosis agent. These results suggested that medicinal and wild plants could be the potential source of thrombin inhibitor.

• BMB Reports
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• 제45권7호
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• pp.390-395
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• 2012

Oleanolic acid (OA), a triterpenoid known for its anti-inflammatory and anti-cancer properties, is commonly present in several medicinal plants but its anticoagulant activities have not been studied. Here, the anticoagulant properties of OA were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT), fibrin polymerization as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed OA prolonged aPTT and PT significantly and inhibited thrombin catalyzed fibrin polymerization. In addition, OA inhibited the activities of thrombin and FXa and inhibited the generation of thrombin or FXa in human endothelial cells. OA also inhibited TNF-${\alpha}$-induced tissue factor expression on human endothelial cells. In accordance with these anticoagulant activities, OA showed an anticoagulant effect in vivo. These results indicate that OA possesses antithrombotic activities and suggest that daily consumption of a herb containing OA may be preventing thrombosis in pathological states.

• BMB Reports
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• 제46권10호
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• pp.484-489
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• 2013

Piperlonguminine (PL), an important component of Piper longum fruits, is known to exhibit anti-hyperlipidemic, antiplatelet and anti-melanogenic activities. Here, the anticoagulant activities of PL were examined by monitoring activated-partial-thromboplastin-time (aPTT), prothrombin-time (PT), and the activities of thrombin and activated factor X (FXa). The effects of PL on the expressions of plasminogen activator inhibitor type 1 (PAI-1) and tissue-type plasminogen activator (t-PA) were also tested in tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) activated HUVECs. The results showed that PL prolonged aPTT and PT significantly and inhibited the activities of thrombin and FXa. PL inhibited the generation of thrombin and FXa in HUVECs. In accordance with these anticoagulant activities, PL prolonged in vivo bleeding time and inhibited TNF-${\alpha}$ induced PAI-1 production. Furthermore, PAI-1/t-PA ratio was significantly decreased by PL. Collectively, our results suggest that PL possesses antithrombotic activities and that the current study could provide bases for the development of new anticoagulant agents.