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Loperamide hydrochloride as antineoplastic component and uses thereof

A technology of loperamide hydrochloride and antineoplastic drugs, which is applied in loperamide hydrochloride as an antineoplastic drug component and its application field, and can solve the problems of large drug resistance, toxic side effects, and easy drug resistance

Inactive Publication Date: 2009-03-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, while traditional cytotoxic drugs kill tumor cells, they also act on normal cells, causing serious side effects, and tumor cells are prone to develop drug resistance under the pressure of survival.
In recent years, although the molecular targeted drugs developed rapidly in recent years, although the targets are clear and specific, there are also a series of reports of off target effects, and a single drug has a high probability of developing drug resistance in the human body.

Method used

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  • Loperamide hydrochloride as antineoplastic component and uses thereof
  • Loperamide hydrochloride as antineoplastic component and uses thereof
  • Loperamide hydrochloride as antineoplastic component and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1, the inhibitory effect of loperamide hydrochloride on cell proliferation

[0022] Loperamide hydrochloride was dissolved in dimethyl sulfoxide (DMSO) or methanol to prepare a mother solution with a concentration of loperamide hydrochloride of 50 mM for subsequent use.

[0023] Dilute logarithmic growth phase liver cancer cells SMMC7721 to 2×10 5 cells / ml, inoculate in a 96-well culture plate, add 0.1ml to each well, place at 37°C, 5% CO 2 and saturated humidity incubators. After the cells are fully adhered to the wall, carefully aspirate and discard the supernatant. Dilute the mother solution with RPMI1640 culture medium, so that loperamide hydrochloride can reach different final concentrations (100uM, 50uM, 25uM, 12.5uM, 6.25uM), each concentration is parallel to 5 wells, and 0.1ml is added to each well, and the blank control group is added. The same amount of diluted culture solution was placed in an incubator for 24 hours (the culture conditions were ...

Embodiment 2

[0030] Embodiment 2, PI single staining assays the cell apoptosis induced by loperamide hydrochloride

[0031] Take the lung adenocarcinoma cell SPC-A1 in the logarithmic growth phase, and use 2×10 5 The density of each / ml is inoculated on the bottom area of ​​25cm 2 culture flask at 37°C, 5% CO 2 and the incubator of saturated humidity are cultivated, after the cells are fully adhered to the wall, dilute the loperamide hydrochloride mother solution (50mM) in the embodiment 1 with 1640 medium, so that the final concentration of loperamide hydrochloride in the culture bottle is respectively 0, 12.5uM, 25uM, 50uM, continue to culture for 24h (culture conditions as above). Add 200ul of 0.25% trypsin to digest the cells, centrifuge at 1000rpm for 6min, collect the culture supernatant and the adherent digested cells, wash twice with PBS, and centrifuge to remove the supernatant. Add pre-cooled 70% ethanol, and fix the cells at 4°C for more than 3h or overnight. Centrifuge, remo...

Embodiment 3

[0033] Example 3, FITC-Annexin V / PI double staining detection of early cell apoptosis

[0034] Take the lung adenocarcinoma cell SPC-A1 in the logarithmic growth phase, and take 2×10 5 The density of each / ml is inoculated on the bottom area of ​​25cm 2 culture flask at 37°C, 5% CO 2And the incubator cultivation of saturated humidity, after the cells are fully adhered to the wall, dilute the loperamide hydrochloride mother solution (50mM) in the embodiment 1 with 1640 medium, so that the final concentration of loperamide hydrochloride in the culture bottle is 37.5 uM, continue to culture (culture conditions as above), after 0h, 6h, 12h, 24h respectively, add 0.25% trypsin 200ul to digest the cells, centrifuge at 1000rpm for 6min, collect the culture supernatant and adherent digested cells, wash with PBS 2 times, centrifuge to remove the supernatant. According to the instructions of the FITC-Annexin V / PI double staining kit, FITC-Annexin V and PI were added for staining. At ...

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Abstract

The invention discloses a new application of loperamide hydrochloride, which is applicable to the preparation of antineoplastic drugs. Loperamide hydrochloride has great inhibition effect on the proliferation of tumor cells. Loperamide hydrochloride can induce obvious apoptosis of tumor cells.

Description

technical field [0001] The present invention relates to the application of loperamide hydrochloride, especially the application as an antitumor drug. Background technique [0002] Malignant tumors, including carcinoma and sarcoma, are common and intractable diseases that endanger human health. With the development of science and technology, people have developed a variety of chemotherapeutic and biotherapeutic drugs. Biotherapeutics that can be used alone or in combination with chemotherapy include cytokines, hematopoietic growth factors, adoptive immunotherapy, somatic cell therapy, epidermal growth factor tyrosine kinase inhibitors, angiogenesis inhibitors, matrix metalloproteinase inhibitors, Regulators of cell signaling pathways, antisense therapy, farnesyl converting enzyme inhibitors, multi-target antifolate drugs, tumor necrosis factor-related apoptosis-inducing ligand, somatostatin, monoclonal antibody targeted therapy, tumor vaccine therapy , gene therapy, etc. I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/451A61P35/00
Inventor 宫兴文徐宇虹
Owner ZHEJIANG UNIV
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