| BACKGROUNDStypticum is one of vital and indispensable drugs for saving lives or treatingdiseases in the clinic. At present, according to their clotting mechanism, the popularstypticums used clinically can be divided into several groups. The first is Batroxobin,a kind of thrombin-like enzyme from snake, such as Reptilase, Hemocoagulase atrox,etc. The second is a kind of styptic working on the thrombin in blood, such asVitamin K, human prothrombin complex made by.means of freeze drying gelsiccation,thrombin, etc. Antifibrinolytic stypticum is the fourth, such as tranexamic acid andaprotinin. Some stypticums can influence the capillary permeability, such asCarbazochrome Sodium Sulfonate, Etamsylate, etc. Chinese drugs are also involvedin these groups. Among these styptic groups, thrombin-like drugs are used mostcommonly, which are mainly divided into three types: thrombin-like enzyme fromsnake, thrombin and hemocoagulase. According to statistic, thrombin-like enzymefrom snake were used in domestic usually.Some studies present that snake venom contains rich blood coagulatingcomponents. Related active factors of these components aimed at every bloodcoagulation factor in the human intrinsic coagulation system can also be found in thesnake venom. Some elements in the venom of crotalinae, viperinae and elapidase canaffect one or multi key enzymes during the process of coagulating enzyme cascadereaction in blood. Some can activate the blood coagulation factors, such as FⅤ, FⅦ,FⅨ, FⅩand FⅡ; or make fibrin (batroxobin) coagulate directly. The density andspeed of their blood coagulating have big difference, which may be related to their different clotting mechanisms. They can work on one or multi links during the bloodcoagulation path, to make blood clot. Since 1976, when FX blood coagulationelement was found and purified from Russell’s viper by Kisiel et al, multiplecomponents affecting blood coagulation system have been purified and characterizedfrom snake venom.In 1936, a kind of enzyme preparation of high purity was separated and purifiedfrom Batroxobin venom, according to the method which was firstly introduced byProf. V1 Klobusitzky and applied for patent. It is composed of two elements:Batrox-obin, also called as Batroase or Reptilase, and a small amount ofphospholipid-depending factorⅩactivator (FXa). Within the series of thrombin-likeenzymes from snake, the most popular is from the ophiotoxin of bothrops atrox(Batroxobin, Reptilase, made by Solco Basle Ltd, Switzerland), and of Cal losellasma rhodostoma (Knoll AG pharmaceutical factory, Ludwigshafen, Germany),whose representing products is Reptilase.Because of many advantages of immediateeffect, high performance, prolonged action, safe, convenience and impregnability byinhibitor of plasma thrombin, Reptilase has been widely used to treat and prevent allkinds of hemorrhagic diseases. In China, from August of 1988 or in 1992, XiaoChanghua or Guan Jinhua et al has managed to separate the thrombin-likecomponents having the activities of blood coagulation and thrombolysis fromAgkistrodon acutus for many times. But these components were difficult tocharacterize and quantify due to the unavailability of purified products. Since 1993,Tang Songshan et al has started on studying the thrombin-like components fromAgkistrodon acutus venom. In the March of 2001, they obtained Agacutin and made aquality control guideline according to the international standard. At present, only theinjected thrombin-like enzyme from Agkistrodon blomhoffii ussurensis of Changbaimountain (brand name: hemocoagulase atrox) going on the market is investigated anddeveloped by our country. Therefore, it is significant to open up more hemostasisreagents with the advantages of immediate effect, high performance and less sideeffect, for breaking the present situation on the dependence of import and protectingthe national drug factory. Haemocoagulase Acutus for Injection is extracted from the venom ofAgkistrodon acutus, which is unique in China. Its physiological function is similar asthat of Batroxobin from Bothrope jararoca venom, which was demonstrated to havegood hemostasis effect by animal experiments. The national first-class new drug ofinjected thrombin-like enzyme from Agkistrodon acutus, which was co-applied byGuangdong shunfeng pharmaceutical co-ltd and Zhongda tianyi bioengineeringtechnology co-ltd, was separated and purified from the Agkistrodon acutus in theSouthern China area. The results from preclinical study confirmed it a kind of safeand effective hemostasis drugs due to the feasibility of its quality control, definitehemostasis effect and extremely low toxic reaction. And it obtained the approval fromthe state drug administration for the first-phase clinical trial in the December of 2002.From December of 2003 to April of 2004, the first-phase clinical trial of this drug hasbeen carried out at the base of national drugs clinical study in Daping hospital, ThirdMilitary Medical University.ObjectiveTo investigate the clinical effect and safety of a single injection of 1U and 2UHaemocoagulase Acutus for Injection for the hemostasis at surgical incision. Basedon the snake resources in China, it is aimed to study and develop a kind of injectedthrombin-like enzyme from Agkistrodon acutus similar as Reptilase, for breaking thepresent situation depending on import completely.Methods and Results1. The clinical application study atⅡa phaseThe experiment was designed according to the principles of random, single-blindcontrol. 45 people were randomly divided into the three groups: experimental group1U, 2U and control group (15 cases per group). The cases were divided intoexperiment group and control group by the professional biomedicine expertsaccording to the random data generated by test centre, and every case was put into thesealed letter randomly. The drug number was written on the letter and the categorygroup was labelled in the letter, which was prepared for every case. Those caseswhich were distributed randomly according to the circa order and the drug number in the principal from little to big. And fit those cases into experimental group andcontrol group as the number labelled in the letter.The moderately elective operation of the abdomen was selected. In the 1U and2U experimental groups, 1U and 2U Haemocoagulase Acutus for Injection wereintravenously injected at 30min before surgery. No hemostatic was used in controlgroup before operation. The incision was cut into the tissue before aponeurosis ofinvestment. Hemorrhage time was recorded by using the adjusted electronicseconds-counter. Hemorrhage volume was calculated the weight difference of gauzebefore and after it was placed in the surgical incision with a electronic balance. Thelength and depth of incision were also examined and its area was then accounted.Thus hemorrhage volume per mm~3 could be obtained.Before medication, the incision of every case was observed for 10 min.Hemospasia was carried out at ld and 3d after the surgery to examine some bloodcoagulation indexes, such as PT, TT, APTT, FIB and PLG, for evaluating the effectof drug on the blood coagulation. Other things still needed to be investigated beforemedication and at 3d after surgery, including vital sign record, electrocardiogram,routine blood and urine examination, liver and renal function, electrolyte, total bloodviscosity, systemic and local reaction et al, for evaluating the safety of the drugs andcalculating the rate of ill occurrence.In the experimental groups of two doses (1U and 2U), hemorrhage volume andhemorrhage volume per unit area of the surgical incision were both less than those incontrol group, and the difference has the statistical significance (P＜0.05). But thehemorrhage time in both doses was also less, while the difference was not obvious.The above three indexes in 2U dose group were all less than those in 1U group andthe differences were also not evident (P＞0.05). There were no significant differenceof coagulation indexes, routine blood and urine examination, liver and renal functionamong three groups. Most ill occurrence happened in the experimental process werethe abnormal but slight changes about the routine blood examination and bloodcoagulation indexes. There were not obvious different among the three groups.2. The clinical application study atⅡb phase This experiment was designed according to the principles of random,double-blinded and placebo control. 36 cases were randomly divided into 2Uexperimental group and placebo group (18 cases per group). According to the layersof test centre, SAS statistic soft was used to produce the random numbers and theseeding number was then decided. 120 cases with different diseases accepted therandom arrangement and labelled with the number from 001 to 120. After that therelated therapy were allotted. According to 20 percent of total cases, every studyingcentre prepared spare medication numbers and the associated drug boxes, in order tosupply the loosing cases. If the loosing cases happened during the experiment, thespare number could be used after all the cases finished entering groups, till supplyingthe loosing cases and control cases.The moderately elective operation of the abdomen was selected. In theexperimental groups, 2U Haemocoagulase Acutus for Injection were intravenouslyinjected at 30min before surgery. Placebo was used in control group before operation.The incision was cut into the tissue before aponeurosis of investment. Hemorrhagetime was recorded by using the adjusted electronic seconds-counter. Hemorrhagevolume was calculated the weight difference of gauze before and after it was placedin the surgical incision with a electronic balance. The length and depth of incisionwere also examined and its area was then accounted. Thus hemorrhage volume permm~3 could be obtained.Before medication, the incision of every case was observed for 10 min.Hemospasia was carried out at 1d and 3d after the surgery to examine some bloodcoagulation indexes, such as PT, TT, APTT, FIB and PLG, for evaluating the effectof drug on the blood coagulation. Other things still needed to be investigated beforemedication and at 3d after surgery, including vital sign record, electrocardiogram,routine blood and urine examination, liver and renal function, electrolyte, total bloodviscosity, systemic and local reaction et al, for evaluating the safety of the drugs andcalculating the rate of ill occurrence.The results suggested hemorrhage time, hemorrhage volume and hemorrhagevolume per unit area of the surgical incision in experimental groups were all less than those in control group, and the differences were significant (P＜0.05). There were PTprolong lightly in the two groups at 10min and 1d after the surgery, and thedifferences were no significant(P＞0.05). There were FIB step up gently in the twogroups at 3d after the surgery, and the differences were no significant, too (P＞0.05).There were no obvious differences of vital sign record, electrocardiogram, routineurine examination, liver and renal function between the two groups onbefore-and-after medication. Some routine blood examination index have greatchange in the two groups, but the differences were no significant. The differencewere no significant on the type and rate of ill occurrence in experimental group andcontrol group.Conclusion:1. A single injection of 1U and 2U Haemocoagulase Acutus for Injection for thehemostasis at 30min before the moderately elective operation of the abdomend,Whichboth have the definite haemostasis effect. And the dose of 2U is more obvious than1U.2. A single injection of 1U and 2U Haemocoagulase Acutus for Injection for thehemostasis at 30min before the moderately elective operation of the abdomend,Whichboth have no significant effect on hemoagglutination function index.3.A single injection of 1U and 2U Haemocoagulase Acutus for Injection for thehemostasis at 30min before the moderately elective operation of the abdomend,Whichboth have no significant effect on vital sign record and electrocardiogram.4. A single injection of 1U and 2U Haemocoagulase Acutus for Injection for thehemostasis at 30min before the moderately elective operation of the abdomend, Whichhave no significant effect on Liver and Renal function index. |
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