Abstract: A purification process of venom blood coagulation enzyme for injection comprises, venom dissolution, compatible chromatography segregation to obtain the coarse solution of batroxobin and thrombokinase, hyperconcentration, ion exchange chromatography purification to obtain the pure solution of batroxobin and thrombokinase, hyperconcentration, gel chromatography to obtain the refined pure solution of batroxobin and thrombokinase, mixing the refined pure solution of batroxobin and thrombokinase according to a finite proportion, filling right amount of excipient, stabilizing agent, and filtering, loading, freeze drying, rolling cap, finally obtaining the venom blood coagulation enzyme for injection

Abstract: A Batroxobin prepared by gene recombination method is disclosed, which can be used as the main component of anastaltic or directly as norfibrosis medicine. Its synthesis, expression and purification of expressed product are also disclosed.

Abstract: Objective This study was designed to use urokinase (UK) in combination with batroxobin in thrombolytic therapy so as to see whether batroxobin(DF-521) would be effective for neuroprotection. Methods The model of right middle cerebral artery occlusion (MCAO) in male SD rats was established. 120 rats were randomized into 9 groups, namely control group, sham control group, and groups that were treated with batroxobin and urokinase together or separately. Each group comprised 15 rats. Intracranial bleeding, infarct volume ratio and neurological function were observed. Results Intracranial bleeding was found in 5 rats of the UK 5000 U/kg group, in 4 rats of the UK 5000 U/kg (2 h) + DF-521 5 BU/kg (2 h) group, and in only 1 rat of the UK 5000 U/kg (2 h) + DF-521 5 BU/kg (1 h) group. Cerebral infarct volume ratio was obviously reduced in 5 BU/kg batroxobin group. No difference was observed in neurological deficit scores. Conclusion 5000 U/kg urokinase increased the risk of intracranial hemorrhage in rat MCAO model. Batroxobin either used separately or in combination with urokinase would not increase the risk of intracranial hemorrhage in rat MCAO model.

Abstract: To investigate the inhibition effect and mechanisms of Batroxobin, a thrombin-like enzyme prepared from snake venom,on tumor growth and metastasis.The models for subcutaneous tumor growth, experimental metastasis and spontaneous metastasis were established by using B16-BL6 melanoma and U14 cervical carcinoma cell lines all with high potential of metastasis.The level of plasma fibrinogen was detected by using enzyme-linked immunosorbant assay (ELISA).The fibrinogen deposition in tumor tissues was determined by immunohistochemical analysis.Batroxobin 40BU/kg significantly inhibited subcutaneous tumor growth of B16-BL6 melanoma and U14 cervical carcinoma,with growth inhibition rates of 85.7%and 77.4%,respectively.In the experimental metastatic model of U14,a significant reduction in the number of pulmonary metastatic lesion was observed in Batroxobin-treated mice when compared with control mice.The number of metastatic lesion in control mice was 133, while the number reduced to 24, 24.5, 26 and 52 after Batroxobin treatment with various regimens.In the spontaneously metastatic model, Batroxobin not only inhibited primary (intramuscular) U14 tumor growth, but also significantly decreased pulmonary micrometastatic lesions and inhibited lymphatic metastasis.Immunohistochemical analysis showed that Batroxobin greatly decreased the deposition of fibrinogen into the extracellular matrix of B16-BL6 tumors.Furthermore,Batroxobin injections were well tolerated; having no harmful effects on body weight and bone marrow or on the general well-being of the animals.[Conclusion]Our data suggest that Batroxobin can inhibit tumor growth and metastasis without bone marrow depression,and will be a promising agent in cancer therapy.