Use of tumor vascular disrupting agent DMXAA in preparation of antiplatelet and antithrombotic drugs
An anti-thrombotic drug and anti-platelet technology, which can be used in drug combinations, cardiovascular system diseases, blood diseases, etc., and can solve problems such as DMXAA, a tumor vascular destroying agent, which has not yet been seen.
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Embodiment 1
[0030] Example 1. The Inhibitory Effect of DMXAA on U46619-Induced Human Platelet Aggregation
[0031] The purpose of this example is to prove that DMXAA can inhibit human platelet aggregation induced by U46619
[0032] In this example, platelets were prepared: the platelets came from healthy volunteers who had signed the informed consent form, and the volunteers had not taken any antiplatelet drugs such as aspirin and clopidogrel within 20 days before blood collection; the anticoagulant was ACD (85mmol L -1 sodium citrate, 71.38 mmol L -1 citric acid, and 27.78mmol L -1 Glucose); centrifuged at 300g for 20 minutes to obtain platelet-rich plasma, after taking the supernatant, centrifuged at 900g for 10 minutes to obtain platelets, and finally resuspended with Tyrode's buffer (138mmol L -1 NaCl, 2.7mmolL -1 KCl, 2 mmol L -1 MgCl 2 ,0.42 mmol L -1 NaH 2 PO 4 ,5mmolL -1 Glucose, 10mmol L -1 HEPES, 0.2% bovine serum albumin, and 0.02unit mL -1 apyrase, pH 7.4),...
Embodiment 2
[0039] Example 2. The inhibitory effect of DMXAA 300 μM on human platelet aggregation induced by other various agonists
[0040] The purpose of this example is to demonstrate that DMXAA can inhibit the aggregation of human platelets induced by arachidonic acid, ADP, collagen or rystokinesis.
[0041] In this example, the preparation of platelets and the assay method of platelet aggregation are the same as in Example 1.
[0042] The results are shown in Table 2. DMXAA at 300 μM significantly inhibited the aggregation of aspirin-untreated human platelets induced by arachidonic acid, ADP, collagen or ristomycin (these four agonists were purchased from Chrono-Log Company) .
[0043] Table 2 shows that DMXAA inhibits the aggregation of aspirin-untreated human platelets induced by arachidonic acid, ADP, collagen or rystofamicin (mean ± standard error) (n=3).
[0044] Table 2
[0045]
Embodiment 3
[0046] Example 3.DMXAA inhibits TXA 2 generate test
[0047] The purpose of this example is to prove that DMXAA can inhibit TXA 2 generate.
[0048] In this example, human washed platelets without aspirin treatment were prepared, and the final concentration was adjusted to 2.5×10 8 cells / ml, with 500 μl / tube as the reaction system, on the platelet aggregometer, add drugs and incubate for 3 minutes, then add agonists to stimulate the reaction until the 8th minute, add 1 / 6 volume of indomethacin (0.55mM) and EDTA (22mM) terminated the reaction, quickly placed on ice, centrifuged at 12000g 4°C for 3 minutes to remove platelets, and the resulting supernatant was stored at -80°C; due to TXA 2 Extremely unstable, quickly transitions to TXB 2 , thus detecting TXB 2 TXA 2 Generate; this embodiment adopts the TXB prepared by the thrombus room of Soochow University School of Medicine 2 Enzyme-linked immunoassay kit for detection of TXB 2 Content, specific steps refer to its inst...
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