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An anti-tumor combination drug

An anti-tumor and drug technology, applied in the field of biotherapeutic drugs, to achieve the effect of improving killing, good safety and improving killing ability

Active Publication Date: 2019-09-17
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no report on the enhancing effect of bisphosphonate drugs on the killing ability of CIK cells.

Method used

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  • An anti-tumor combination drug
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1 Induction of CIK cells

[0048] (1) Aseptically collect 50ml of peripheral blood from normal people, anticoagulate with heparin, send it back to the laboratory, and perform follow-up operations in a sterile ultra-clean bench.

[0049] (2) Dilute the blood sample with PBS buffer solution (0.01M, pH 7.4, the same below) equal to the volume of the blood sample. Add human lymphocyte separation medium (Shenzhen Dakowei Biological Engineering Co., Ltd.) equal to the volume of the undiluted blood sample in a sterile centrifuge tube, and then slowly add the blood sample diluted with PBS buffer on top of the separation medium, avoiding the blood sample from mixing as much as possible. The separation solution was mixed to keep the interface of the two liquids clear, wherein the volume ratio of the separation solution, undiluted blood sample and PBS was 1:1:1.

[0050] (3) Centrifuge at 800 g with a horizontal rotor for 20 min at room temperature. After centrifugation,...

Embodiment 2

[0056] Example 2 examines the killing ability of CIK and zoledronic acid alone on prostate cancer cell PC-3.

[0057] The killing effect of CIK cells on tumor cells was detected by MTT method:

[0058] Prostate cancer cell PC-3 (purchased from ATCC, USA) was cultured in F12 medium (Gibco, Life technology) containing 10% FBS (Gibco, Life technology) in a medium containing 5% (v / v) CO 2 , 37°C cell culture incubator.

[0059] PC-3 cells (prostate cancer cell PC-3) were digested with 0.25% trypsin (containing 0.02% (m / v) EDTA (ethylenediaminetetraacetic acid)), counted and inoculated into 96-well plates, 4000 per well cells, the volume of medium (F12 medium) was 100 μL. Place in the incubator for 24 hours until the cells adhere to the wall. Divided into control group, CIK treatment group and zoledronic acid group for experiment.

[0060] Control group: add 100 μL F12 complete medium (containing 10% (v / v) fetal bovine serum (FBS)), without CIK and zoledronic acid;

[0061] CI...

Embodiment 3

[0066] Example 3 detects the enhancing effect of zoledronic acid on the killing ability of CIK cells after pretreatment of target cells.

[0067] In order to detect the increasing effect of zoledronic acid on the killing ability of CIK cells, the killing ability of CIK cells on PC-3 cells was detected after zoledronic acid was pretreated with PC-3 prostate cancer cells by MTT method (the same method as in Experimental Example 2). .

[0068] Prostate cancer cell PC-3 was seeded into 96-well plate at 4000 / well. After the cells adhered to the wall for 24 hours, the culture medium (F12 complete medium) was aspirated, and 100 μL of F12 medium containing 4, 8, and 16 μM zoledronic acid was added for pretreatment for 12 hours, and then CIK cells 20 times the number of target cells were added (ie 80000 / well, volume 100 μL) for 24 hours; treated with zoledronic acid alone and CIK cells alone as a comparison (the method is the same as in Example 2), and then the cell viability was dete...

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Abstract

The invention discloses an antitumor combined medicine. The combined medicine comprises a diphosphonate compound serving as an active ingredient and CIK cells, wherein the diphosphonate compound is at least one of clinical medication etidronate disodium, clodronate disodium, pamidronate disodium, tiludronate disodium, alendronate sodium, neridronate sodium, olpadronate sodium, risedronate sodium, sodium ibandronate, incadronate disodium and zoledronic acid. The method for greatly improving the tumor cell killing capability of CIK cells by utilizing combination of anti-tumor-osseous-metastasis diphosphonate medicines and the CIK cells, so that the number of CIK cells for achieving the treatment effect the same as a conventional method is greatly reduced; and compared with biological treatment of single CIK cells, the combined medicine has the advantages that the relatively high safety of CIK cells can be maintained, and the treatment effect can be remarkably improved.

Description

technical field [0001] The invention belongs to the field of biotherapeutic drugs, and in particular relates to providing an anti-tumor combined drug. Background technique [0002] The incidence of cancer is increasing day by day. According to the World Health Organization, there were about 14 million new cancer cases and 8.2 million cancer-related deaths worldwide in 2012. Cancer is the leading cause of morbidity and mortality worldwide, and the number of new cases is projected to increase by approximately 70% over the next two decades (WTO, World Cancer Report 2014). Although there are more and more treatment methods for cancer and the available drugs are also increasing, most cancers are still incurable and the survival period of patients is short. Therefore, any new safe and effective cancer treatments or means that can improve the effectiveness of existing treatments will have practical implications for the health of cancer patients. [0003] Cytokine-induced killer c...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/17A61K31/675A61P35/00
Inventor 陈填烽曾德龙赵建夫
Owner JINAN UNIVERSITY
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