• Archives of Pharmacal Research
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• 제24권2호
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• pp.109-113
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• 2001

Curdlan is a natural $\beta$-1,3-glucan produced by Agrobacterium biovar 1. In this study, the anticoagulant activity of sulfoalkyl derivatives of curdlan was investigated by carrying out activated partial thromboplastin time (APTT) assay and compared with that of o-sulfonated curdlan. Approximately 100-fold higher concentration of o-sulfonated curdlan than heparin was required to obtain the same level of the clotting time. Anticoagulant activity of curdlan derivatives was dependent on the degree of sulfation in prolonging the clotting time. However, the chain length of the substituent did not play a role in prolonging the clotting time. The curdlan derivatives enhanced thrombin inhibition by mediating through antithrombin III. The inhibition of thrombin by o-sulfonated curdlan was found to be approximately 10-fold weaker than that by heparin.

• 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.

• Applied Biological Chemistry
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• 제12권
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• pp.7-11
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• 1969

Mucor-rennin, the crystalline milk-clotting enzyme, isolated from Mucor pusillus var. Lindt, has an acid protease activity. The optimum pH for the digestion of k-casein is 4.5, while that for hemoglobin digestion is 4.0. The skim milk solution was easily clotted acidic solution than alkalin solution, and the milk clotting activated by Ca ion. The enzyme was heat stable against heat from pH 4.0 to 6.0 but was more stable at pH 5.0. The activity of the enzyme at pH 5.0 did not decrease at 30 C for 15 days and the activity was not effected by sodium propionate and salicilic acid. Therefore, the enzyme of liguid type could store for a long time and could be transported from Erzyme production Co. to Manufacture of cheese Co. by adding the antiesptic and by adjusting pH to 5.0.

• Journal of Chest Surgery
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• 제33권7호
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• pp.560-564
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• 2000

배경; 아프로티닌(Aprotinin)은 nonspecific serine protease inhibitor로 개심술에서 항염효과와 혈액 응고효과를 갖고 있다. 또한 아프로티닌은 심폐기 작동 중에는 항트롬빈(antithrombin) 효과가 있다. 본 연구는 아프로티닌이 헤파린(heparin)사용 양 및 ACT(activated clotting time) 에 어떠한 영향을 미치는 가를 알아보기 위하여 시행하였다. 대상 및 방법; 1998년 12월 1일부터 1999년 11월 30일 까지 본원에서 개심술을 받은 환자 중 연령이 18세 이상인 82명을 대상으로 연구하였다. 환자들은 아프로티닌을 사용한 군(Group A)과 사용하지 안흥ㄴ 군(Group C)으로 나누어 연구하였다. 모든 환자에서 수술 전 체중, 신장, 체표면적, pump time, ACC time 등을 조사하였다. ACT의 측정 시기는 헤파린 공급 전, 공급 후 20분, 40분, 60qs과 프로타민 공급 후 20분 등이었다. 또한 ACT 변화 정도를 알아보기 위하여 헤파린 공급 전과 공급 후 20분, 공급 후 40분과 20분 사이의 차이를 조사하였다. 결과; 연구 대상 환자들이 굴변 특징에서는 연령, 펌프시간 및 대동맥차단 사간에서 모두 A군에서 증가된 소견을 보였다. (p<0.05). 헤파린 공급 전과 프로타민 공급 후의 ACT는 두 군에서 차이가 없으며 헤파린 공급 후 20분(607$\pm$22.3, 525$\pm$169초), 40분(889$\pm$315, 546$\pm$103초), 60분 (748$\pm$310, 472$\pm$115 초)에 측정한 ACT는 모두 A군에서 증가된 소견을 보였다. (p<0.05), 헤파린 공급 전과 궁급 후 20분 사이의 ACT 차이는 A한 군에서 증가된 소견을 보였고(p<0.05), 헤파린 공급 후 40분과 20분 사이의 ACT의 차이 또한 A군에서 증가된 소견을 보였다. (p<0.05). 두 군간의 총 헤파린 및 프로타민 사용 양에는 차이가 없었다. (p>0.05). 결론; 결론적으로 아프로티닌은 본 연구에서 CPB time을 고려해 볼 때 헤파린사용 양을 의미있게 줄여 줄 수 있을 것으로 생각되며 또한 ACT를 증가시켜 주기 때문에 추가적인 헤파린 감량이 가능할 것으로 생각된다.

• 대한본초학회지
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• 제26권4호
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• pp.139-147
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• 2011

Objectives : The aim of this study is to research an anti-thrombus effect by Diospyros Kaki Calyx. Methods : The healthy human plasma were gained and used 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 (orally administrated with distilled water 5ml/kg) and two experimental group treated with extract of diospyros kaki calyx (EKC). Experimental rats were orally 600 mg/kg concentration of EKC and 200 mg/kg concentration of EKC. After an hour from administration, we anesthetized rats and made arteriovenous (AV) shunt rat models to study weight of thrombus, took whole blood to study content of thromboxane B2 and blood clotting time. Results : In vitro, EKC significantly increased inhibitory activity of FXa, prothrombinase compared with intact control group ($^*P$ <0.05). PT and aPTT were increased in EKC treated (600 mg/kg) group compared with intact control group ($^*P$ <0.05). In vivo, blood clotting time of experiment group treated with EKC 600 mg/kg were significantly increased compare with that of intact control group (p<0.05) and content of thromboxane B2 was significantly decreased in group treated with EKC 600 mg/kg in serum. The weight of thrombus were significantly reduced in group treated with EKC 600 mg/kg compared with intact control group (p<0.05). But in vivo experiment study, those parameters of group treated with EKC 200 mg/kg were relatively decreased compared with those of intact control group without statistical significance. Conclusions : EKC has an antithrombic activity because of inhibition internal course such as FXa and prothrombin. And EKC inhibited a hole blood clotting in vivo experiment by low content of thromboxane B2.

• Journal of Chest Surgery
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• 제13권4호
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• pp.346-355
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• 1980

It has been proposed that wide individual variation in response to heparin be not considered in the conventional set protocol for the control of heparin and protamine during extracorporeal circulation. In this paper, two protocol of heparin and protamine therapy were compared to assess the role of the Activated Clotting Time [ACT] in relation to heparin, protamine, and postoperative blood loss and transfusion. The study groups consisted of the 31 patients [adults 15 and children 16] anticoagulated with the conventional heparin protocol and the 31 patients [adults 15 and children 16] anticoagulated with ACT protocol during extracorporeal circulation. In the conventional heparin protocol, two mg of heparin per kg was administered initially with an additional 0.75 mg of heparin per kg every 30 minutes of extracorporeal circulation, and reversal was accomplished with protamine in a dose of 1.5 times the total milligram of heparin. In the ACT protocol, two mg of heparin per kg was administered initially with an additional dose of heparin enough to reach an ACT of 480 seconds [within safe zone 300 to 600 seconds] from the patient`s dose response curve every 1 hour of extracorporeal circulation, and reversal was done with protamine in a dose of 1.3 times the milligram of the residual heparin. The results were summarized as follows. After a dose of 2 mg per kg of heparin, the patient`s ACT varied from 240 to 600 seconds in adults and from 240 t~ 660 seconds in children. In the ACT group the total amount of heparin administered was markedly reduced when compared to the conventional group, and less protamine was required to neutralize heparin. The dose of heparin administered decreased from 7.07 [SE 0.42] mg/kg of the conventional group to 4.92 [SE 0.32] mg/k8 of the ACT group in adults and from 10.17 [SE 1.15] mg/kg to 5.23 [SE 0.24] mg/kg in children, which represent 30.4% and 48.6% decrease respectively. The dose of protamine administered for reversal decreased from 10.6 [SE 0.63] mg/kg of the conventional group to 3.35 [SE 0.35] mg/kg of the ACT group in adults and from 15.7 [SE 1.70] mg/kg to 3.26 [SE 0.27] mg/kg in children, which represent 68.4% and 79.2% respectively. The ratio of protamine to heparin administered in the conventional group was 1.50:1 in adults and 1.54:1 in children, but in the ACT group 0.68:1 in adults and 0.62:1 in children. Postoperative blood loss and transfusion revealed no statistically significant difference between the two groups. Although six patients in the conventional group and one in the ACT group needed re-exploration for continuous hemorrhage, no case of generalized oozing was encountered, and in each case a definite bleeding site was identified. Author would like emphasizing the value of the ACT protocol in controlling heparin and protamine administration during extracorporeal circulation.

• Natural Product Sciences
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• 제8권2호
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• pp.66-70
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• 2002

The in vitro anticoagulant and fibrinolytic activities of crude extracts from insects were evaluated in order to find effective therapeutic drugs for the treatment of myocardial and cerebral thrombosis. We prepared three types of extracts (water, methanol and ethylacetate) from 28 insects for use as raw materials for the activity assays. The fibrinolytic activity was tested using the fibrin plate method and the activated partial thromboplastin time and thrombin time were measured for blood clotting activity. With regards to the fibrinolytic system, water extracts of six kinds of insects displayed a remarkable level of activity with a plasmin-like action. The water extracts of [Catharsius molossus, Eupolyphaga sinensis, Huechys sanguinea, Mantidis $o\ddot{o}theca$, Mimela splendens, and Polistes mandarinus (Vespae Nidus)] exhibited the activity. On the other hand, the methanol extracts did not display any fibrinolytic activity. In terms of the coagulation system, an aqueous extract of silkworm Tongchunghacho (Paecilomyces japonica), Oxya japonica japonica and Buthus martensi (Scorpion) increased the clotting time significantly longer (181 times) than the control. These results suggest that crude drugs from insects are useful sources for the development of new drugs for use in treatments involving blood coagulation and fibrinolysis.

• 한국식품영양과학회지
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• 제33권8호
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• pp.1262-1267
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• 2004

한약재의 재료로 사용되는 옥발에서 항응고물질(anticoagulant)을 추출하여 정제하였다. 정제된 항응고물질의 분자량은 135 kDa으로 측정되었고 옥발로부터 정제된 항응고물질의 회수율은 11%이고, 24배 농축되었다. 옥발의 항응고물질은 열에 안정하고 비특이적 단백질 가수분해 효소 처리 후에도 활성을 유지하였으나, periodate 산화 처리시 현저하게 활성이 소실되어 항응고물질의 성분은 당류인 것으로 추정된다. 옥발의 항응고물질을 thin layer chromatography(TLC)와 Bio-LC로 구성당을 분석한 결과 galactose, glucose, mannose, glucosamine, galactosamine, fucose로 구성되어있고, IR을 통하여 확인된 OH, N-H bond가 확인되어 항응고물질이 중성당과 아미노당으로 구성된 것을 뒷받침해주고 있다. 옥발의 항응고물질은 thrombin time(TT)에 가장 민감하고, TT에서 항응고 활성을 2배, 3배 지연시키는 농도는 60 nM와 88 nM이었다. Prothrombin time(PT)에도 민감하였지만, snake venom activity와 activated partial thromboplastin time(APTT)의 clotting activity에는 상대적으로 별 영향을 끼치지 않았다. 옥발의 항응고물질을 정제된 fibrinogen을 이용하여 thrombin과의 반응 시 fibrinogen의 농도가 증가함에 따라 응고 저해 정도가 감소하여 thrombin에 대해fibrinogen과 결합자리를 경쟁하고, 이 때 Ki 값은 23 nM이다.

• Current Research on Agriculture and Life Sciences
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• 제6권
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• pp.157-162
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• 1988

활성화시킨 Succinylaminopropyl glass beads에 glutaraldehyde을 사용하여 고정화시킨 Mucor spp L42 응유효소를 치즈 커드 형성과정에 이용하고져 고정화 효소의 안정성, 재이용성, 반응의 조건, 반응조의 형태등에 대하여 탈지유(pH 5.6, $8^{\circ}C$)를 사용하여 반응시킨후 $30^{\circ}C$ 항온수조내에서 커드의 형성 상태를 조사하였던바 다음과 같은 결과를 얻었다. 1) 고정화 Mucor spp L42 응유효소를 0.2M phosphate buffer(pH 4.6)에 0.06% sodium azide을 첨가한 액에 침지한 상태로 $5^{\circ}C$ 이하에서 보존할 경우 3개월 후에도 활력의 80%가 유지되었다. 2) 고정화 Mucor spp L42 응유효소를 탈지유(pH 5.6, $8^{\circ}C$)로 반응시켰을때 반응 초기에는 고정화 효소의 활력이 급격히 감소하였고, 이것은 bead에 결합되는 질소화합물과 효소의 활력간에 관계가 있는 것으로 나타났다. 3) 고정화 Mucor spp L42 응유효소에 bead와 탈지유의 접촉시간을 60 sec/ml로 하여 탈지유(pH 5.6, $8^{\circ}C$)로 연속적으로 반응시켰을때 6시간 이후에도 bead의 활력의 70%가 유지할 수 있었다. 4) Fluidized bed와 shaking fluidized bed을 사용하는 것이 충진반응조 보다 탈지유의 반응속도가 높았다.

• 약학회지
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• 제44권6호
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• pp.558-565
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• 2000

SK-1080 is one of the newly developed orally active nonpeptide angiotensinII $AT_1-receptor$ antagonist that selectively acts at $AT_1$ receptor with high affinity. The cardiac effect on ischemia/reperfusion injury of SK-1080 was compared with those of losartan, a prototype of this class, in isolated rat hearts. Isolated perfused rat heart was pretreated with drug for 10 min and then subjected to global ischemia for 30 min followed by reperfusion with- or without drug for 30 min. The possible additive effect of SK-1080 on the platelet aggregation and coagulation in human blood was also studied. We investigated whether SK-1080 effects the platelet aggregation induced by ADP, a platelet agonist partially dependent on $thromboxaneA_2$. The clotting times in the prothrombin time (PT) and activated partial thromboplastin time (APTT) were also examined in human plasma in vitro as coagulation screening test. SK-1080 improved reperfusion function (LVDP, left ventricular developed pressure; PRP, rate-pressure product) in a dose-dependent manner. SK-1080 reduced ADP-induced platelet aggregation compared with vehicle but less than losartan, and did not affect clotting times.