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Combined application of ErbB2 antibody and Saracatinib in preparation of medicine for treating breast cancer

A drug and antibody technology, applied in the field of biomedicine, to achieve the effect of inhibiting tumor growth

Active Publication Date: 2020-10-16
ZHEJIANG SHIMAI PHARMACEUTICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Herceptin has achieved great success in clinical treatment, only about 30% of breast cancer patients respond when they receive Herceptin for the first t

Method used

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  • Combined application of ErbB2 antibody and Saracatinib in preparation of medicine for treating breast cancer
  • Combined application of ErbB2 antibody and Saracatinib in preparation of medicine for treating breast cancer
  • Combined application of ErbB2 antibody and Saracatinib in preparation of medicine for treating breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1. Preparation and purification of anti-ErbB2 antibody

[0041] Recombinant human ErbB2 extracellular domain (ErbB2-ECD) protein was used as immunogen, and Balb / c healthy female mice homologous to SP2 / 0 were selected as immunized animals. The immune antigen was dissolved in PBS, and mixed evenly with an equal volume of Freund's adjuvant to form a water-in-oil emulsion. Subcutaneous multi-point immunization was used for the first immunization, and the immunization dose was 100 μg / bird. 7-10 days after the initial immunization, use incomplete Freund's adjuvant to mix and emulsify with the same volume of immune antigen, and immunize mice with the same route and dosage. Afterwards, the immunization was boosted several times every 7-10 days, so that the antibody titer of the mouse serum reached more than 10,000. One week after the last booster immunization, blood was collected from eyeballs, and serum titers were determined by ELISA. If the mouse serum titer does ...

Embodiment 2

[0042] Example 2. Identification of mouse anti-ErbB2 antibody subtypes

[0043] The subtype of the anti-ErbB2 monoclonal antibody obtained in Example 1 was identified by ELISA (the kit was purchased from Proteintech). The ELISA plate provided in the kit has been pre-coated with specific antibodies against mouse IgG1, IgG2a, IgG2b, IgG3, IgA, IgM, kappa light chain, lambda light chain, and the anti-NSE purified in Example 2 Antibody samples E1-7E2 and E2-4F6 were added to the sample wells respectively, 50 μl per well, without incubation. Add 1X goat anti-mouse IgA+IgM+IgG-HRP into the sample wells, 50 μl per well, mix gently, and incubate for 1 h. Remove the liquid in the well and add 1XPBST to wash the well 3 times, and absorb excess water with absorbent paper. Add chromogenic solution, 100 μl per well, and develop color for 15 minutes at room temperature in the dark. Add 100 μl stop solution to stop the color reaction. Detect the OD value at 450nm by microplate reader, th...

Embodiment 3

[0044] Example 3. Antibody Humanization

[0045] The positive hybridoma cells obtained in Example 1 were expanded and cultured, and an appropriate amount of cells were used for TRNzol-A + After lysis, each 1ml of TRNzol-A + Add 200 μl of chloroform, vortex for 15 seconds, and let stand for 3 minutes. Centrifuge at 13,000rpm at 4°C for 10 minutes. The cell solution is divided into three layers: the upper colorless aqueous phase, the middle layer and the lower yellow organic phase. Transfer the aqueous phase containing RNA to a centrifuge tube, and add an equal volume of isopropyl to the aqueous phase. alcohol, mix well, and let stand at room temperature for 25 minutes. Centrifuge at 13,000 rpm at 4°C for 10 minutes, discard the waste liquid to obtain the RNA pellet that sinks to the bottom. After washing the RNA pellet twice with 75% ethanol, the RNA was dissolved with DEPC water. Subsequently, the first strand of cDNA was synthesized using the TransScript One-Step gDNA Rem...

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Abstract

The invention relates to a combined application of ErbB2 antibody and Saracatinib in preparation of a medicine for treating the breast cancer. The ErbB2 antibody and the Saracatinib have the effect ofsynergistically inhibiting growth of ErbB2 positive tumors when being combined for use, thereby having a wide application prospect.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the combined application of anti-ErbB2 antibody and Saracatinib in the treatment of breast cancer. [0002] technical background [0003] In recent years, the incidence of clinical cancer in the world, breast cancer ranks first in the incidence of female malignant tumors, and it is second only to lung cancer in the incidence of cancer in the world. Obviously, breast cancer is the largest malignant tumor that endangers women's health, and surgery is the key to treatment. The main means of breast cancer, chemotherapy and radiotherapy are recognized as adjuvant therapy. However, in recent years, with the deepening of research on targeted therapy and anti-tumor methods, as well as the development of molecular pathology and genotyping, monoclonal antibody drugs against breast cancer have been rapidly developed. Wherein trastuzumab (trade name Herceptin, Herceptin) is the first antibody drug...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61K31/517A61P35/00
CPCA61K31/517A61K39/39558A61P35/00A61K2300/00
Inventor 彭菲顾超
Owner ZHEJIANG SHIMAI PHARMACEUTICAL CO LTD
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