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Preparation method of intestinal cancer cell-immunized poultry-obtained IgY antibody-coupled broad-spectrum anticancer drug aiming at intestinal cancer and application of drug in prevention and treatment of intestinal cancer

A technology of antibody-drug conjugates and cancer cells, which is applied in the direction of anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, anti-tumor drugs, drug combinations, etc., and can solve problems such as unsatisfactory curative effects

Inactive Publication Date: 2021-03-05
BEIJING WANHUA BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In anti-tumor therapy, antibody drugs have also become standard therapy, but their efficacy alone is often unsatisfactory

Method used

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  • Preparation method of intestinal cancer cell-immunized poultry-obtained IgY antibody-coupled broad-spectrum anticancer drug aiming at intestinal cancer and application of drug in prevention and treatment of intestinal cancer
  • Preparation method of intestinal cancer cell-immunized poultry-obtained IgY antibody-coupled broad-spectrum anticancer drug aiming at intestinal cancer and application of drug in prevention and treatment of intestinal cancer
  • Preparation method of intestinal cancer cell-immunized poultry-obtained IgY antibody-coupled broad-spectrum anticancer drug aiming at intestinal cancer and application of drug in prevention and treatment of intestinal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment example 1

[0016] Preparation of anti-colorectal cancer IgY antibody

[0017] In vitro culture of colorectal cancer cells, according to 10 8 cells / Chicken immunized once, the immunized part is under the wing endothelium, mixed with adjuvant for the first immunization, and then boosted immunization every 15 days, a total of 4 times, eggs were collected, and the binding activity of IgY antibody to colorectal cancer cells was detected.

[0018] Purify the IgY antibody by water dilution method: dilute the yolk body with 10 times of distilled water, adjust the pH of the distilled water to 5.0 with 0.1% hydrochloric acid, stir for 15 minutes, and leave it at 4°C overnight for natural precipitation of lipoproteins, then ultracentrifuge at 10,000 r / min for 25 minutes Lipids were removed, and the supernatant was collected to obtain the IgY antibody against colorectal cancer.

Embodiment example 2

[0020] Anti-colorectal cancer IgY antibody conjugated to 5-fluorouracil (5-FU)

[0021]

[0022] 1) Weigh 20mg of iodoacetylsuccinimidyl ester (NHSIA) and 40mg of 5-FU to react in the presence of potassium carbonate in DMF for 18 hours, add water, extract with ethyl acetate, and evaporate the organic layer to dryness to obtain 5-FU derivatives, weighing Dissolve 50mg of IgY in 1mL of 0.01mol / L PBS with pH7.4, add 20mg of SPDP, stir and react at room temperature for 5 hours to obtain IgY-PDP solution, then add 40mg of dithiothreitol (DTT) for reduction overnight to obtain HS-IgY, The purified HS-IgY solution obtained by dialysis was reacted with 5-FU derivatives overnight, put into a dialysis bag, dialyzed with 0.01moL / L PBS solution at 4°C for 3 days, and changed the solution 3 times a day to obtain 5-FU-IgY.

Embodiment example 3

[0024] Anti-Colorectal Cancer IgY Antibody Conjugated Capecitabine

[0025]

[0026] Dissolve 40 mg of capecitabine in 1 mL of 0.01 mol / L PBS at pH 7.4, add 17 mg of NaIO4 at room temperature, and stir in the dark for 1 hour at room temperature. Excess NaIO4 is eliminated with glycerol with a final concentration of 0.05 mol / L . Add 1ml to dissolve in 0.15mol / L (pH9.5) K 2 CO 3 The reaction of IgY in buffer was stirred at 37°C for 1 hour to form a Schiff base. Add NaBH4 with a slightly excess molarity than NaIO4 of oxidized capecitabine, and co-bath in a 37°C incubator for 2 hours to stabilize the Schiff's base. After centrifugation, dialyze the obtained product into a dialysis bag after centrifugation, dialyze with 0.01moL / L PBS solution for 3 days at 4°C, and change the solution 3 times a day to obtain the capecitabine-IgY conjugate

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Abstract

The invention aims at providing a novel antibody drug conjugate and a preparation method of an Ig gamma antibody-coupled broad-spectrum anticancer drug. Intestinal cancer cells are taken as an immunogen to immunize poultry to obtain an Ig gamma antibody aiming at the intestinal cancer, and a chicken Ig gamma antibody for resisting the intestinal cancer is modified by a plurality of chemotherapeutic drugs and a plurality of different methods, so that a preparation method of an orally-taken bifunctional antibody is provided. After the novel antibody drug conjugate is orally taken, the novel antibody drug conjugate can be combined with the intestinal cancer, and has cytotoxicity for resisting the intestinal cancer, and the drug resistance and toxic and side effects of the chemotherapeutic drugs are reduced.

Description

[0001] Technical field: The present invention relates to the field of therapeutic bifunctional antibodies, and relates to chicken IgY antibodies and modification methods thereof for anti-colorectal cancer. Background technique: [0002] Colorectal cancer (CRC) is one of the most common malignant tumors in the world, and its incidence rate accounts for the forefront of gastrointestinal tumors. The incidence of colorectal cancer in my country is increasing year by year. In 2008, there were about 1.2 million new cases of colorectal cancer in the world, accounting for about 10% of all new cancers, and the related death toll exceeded 600,000. Colorectal cancer is very common in economically developed countries and regions such as North America, Western Europe, and Australia, with a crude incidence rate of 4 / 10,000, accounting for more than two-thirds of all new cases. In addition, the mortality rate of colorectal cancer is very high, mainly due to frequent postoperative metastasis ...

Claims

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Application Information

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IPC IPC(8): C07K16/30C07K16/02A61K47/68A61K31/513A61K31/7068A61K31/555A61K31/4745A61K31/704A61P35/00
CPCC07K16/3046C07K16/02A61K47/6851A61K47/6809A61K31/513A61K31/7068A61K31/555A61K31/4745A61K31/704A61P35/00
Inventor 万积成
Owner BEIJING WANHUA BIOENG
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