A pharmaceutical composition containing anti-vegf antibody
A composition and drug technology, applied in the field of biomedicine, can solve the problems of ineffectiveness, drug resistance, poor effect, etc., and achieve the effect of improving the general state, improving the curative effect, and reducing toxic and side effects
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Embodiment 1
[0041] Example 1: Preparation of the humanized VEGF monoclonal antibody of the present invention
[0042] Rabbit monoclonal antibodies are produced by hybridoma cell technology. See US Patent: 7,429,487, especially Examples 1-4, for experimental protocols.
[0043] First, the IgG Fc-hVEGF-A (Human VEGF165) fusion protein is prepared by recombinant technology, wherein the IgG Fc sequence is of rabbit origin. Clone the DNA sequence of IgG Fc-hVEGF-A into pTT5 plasmid, transiently transfect the plasmid into HEK 293-6E cell line, culture the cells without serum, collect the culture supernatant, and use Protein A column to purify the transiently expressed IgG Fc- hVEGF-A fusion protein.
[0044] New Zealand white rabbits (New Zealand rabbits) were first immunized with purified IgG Fc-hVEGF-A (as an antigen component) mixed with complete Freund's adjuvant for multi-point subcutaneous injection, and purified protein was administered every three weeks thereafter After mixing with i...
Embodiment 2
[0052] Example 2. Efficacy evaluation of servacizumab and Avastin alone on human colon cancer Ls-174t nude mice xenografts
[0053] Experimental materials and animals: Sevacizumab was provided by Jiangsu Simcere Pharmaceutical Research Co., Ltd.; Avastin was purchased from Genentech Co.; BALB / cA-nude mice, 6-7 weeks, ♀, were purchased from Shanghai SLAC Experimental Animals limited liability company.
[0054] Preparation method: Both sevizumab and Avastin are formulated with physiological saline to the required concentration.
[0055] Experimental procedure: nude mice were subcutaneously inoculated with human colon cancer Ls-174t cells, and after the tumor grew to 60-200 mm3, the animals were randomly divided into groups (d0). See Table 1 for dosage and regimen. The tumor volume was measured 2-3 times a week, the mice were weighed, and the data were recorded. The formula for calculating tumor volume (V) is:
[0056] V=1 / 2×a×b 2
[0057] Where a and b represent length and...
Embodiment 3
[0064] Example 3. Evaluation of curative effect of servacizumab alone or in combination with 5-fluorouracil on tumor inhibition in human colon cancer Ls-1 74t nude mice
[0065] Experimental Materials and Animals : 5-Fu provided by Shanghai Hengrui Pharmaceutical Co., Ltd. See Example 1 for animals and other materials.
[0066] Preparation method : 5-prepared with normal saline.
[0067] Experimental procedure : detailed steps are referring to the experimental procedure of embodiment 1. See Table 2 for dosage and regimen.
[0068] Table 2. Efficacy of servacizumab alone or in combination with 5-fluorouracil (5-Fu) on human colon cancer Ls-174t nude mice xenografts
[0069]
[0070] d0: first administration time; P value refers to compared with the contrast. Control group n=1 2, treatment group n=6.
[0071] IV: intravenous injection; IP: intraperitoneal injection.
[0072] result : The application of 5-Fu alone has a certain effect on Ls-174t, and the tumor in...
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