Cell autophagy-based antitumor drug screening method

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The invention belongs to the field of biological pharmacy, and relates to a screening method for screening candidate combination molecules through cell death caused by combination of a tumor cell autophagy inducer and a micro-molecular compound, screened compound molecules may not have antitumor activity, and have the antitumor activity after the screened compound molecules are combined with the cell autophagy inducer, such as an HDAC inhibitor, so a drug combination scheme can be determined. In the invention, cancer cells cultured in vitro are respectively processed by the tumor cell autophagy inductor and single compounds as an object, the degree of the cultured cell death caused by the compounds and the cell autophagy inducer is determined, and cell autophagy-based antitumor drugs are screened. A result of cell experiments proves that the method can be used to screen the combination drugs in order to prepare antitumor medicine compositions of the HDAC inhibitor and the micro-molecular compound, and the medicinal compositions are suitable for all dosage forms.

The present invention belongs to the field of medical technology, anticancer drugs based screening method involves autophagy; in particular to an increased cancer cells sensitive to small molecule HDAC inhibitor compounds and compound screening methods, as well as small molecule HDAC inhibitors compound antitumor composition.

With the deepening of lysosomal study focused on Christian deDuve containing cytoplasmic vesicles and organelles cell research opens up new areas from the sun, these and related lysosomal vesicles, that since the Liao Jie body (Autophagosome ), in vivo widespread, Christian de Duve also because research lysosomal won the Nobel Prize in Physiology or Medicine 1974 year. Over the past decade, the study found a large self-Jie Liao (Macroautophagy), small self Jie Liao (Microautophagy) and chaperone-mediated from the sun (Chaperone-MediatedAutophagy, CMA) from the sun in the form of three types of cells. Studies have shown that in the process of autophagy occurs, there are many proteins directly or indirectly involved in the different phases of the cell autophagy process, such as I, essential for the production of autophagy Vps34 Beclin, through specific autophagic gene RNA interference or compounds, such as 3-MA (3-methyladenine), hydroxychloroquine, bafilomycin Al 5¾ monensin autophagy inhibition in different ways to inhibit autophagy, but not be able to suppress autophagy induced cell death. Since therefore for autophagy autophagy occurs in tumor development role has attracted more and more attention researchers, many anticancer drugs can induce autophagy produce, autophagy can tolerate chemotherapy and radiotherapy given to cancer therapy, to control tumorigenesis, even appropriate treatment strategies have become a means of cancer prevention.

At present there are many on autophagy inducing agents and autophagy inhibitor screening methods, these methods have focused on the process of autophagy to a certain target molecule, such as a change in fluorescence intensity of labeled LC3 etc. screening autophagy related compounds, these compounds do not necessarily have the ability to kill cancer cells. As we all know, autophagy is a normal physiological process of cell self-regulation, general autophagy can protect cells from death threats, but overactive autophagy can also lead to cell death, such a double-edged sword-like cells from impact bite is difficult to become a discovery tool tumor cell death-inducing agent.

The prior art discloses HDAC (histone deacetylase) and protein acetyltransferase group is a regulatory enzyme histone and non-histone acetylation state, under normal physiological conditions, and their regulation in a relatively balanced state, the cells in the event of conversion, increased HDAC activity so that the original gene expression out of balance, associated with cell proliferation and cell cycle regulation of gene expression imbalance, leading to malignant cell transformation, now has become the field of oncology HDAC a hot topic, some of which have been used as a target for cancer prevention, HDAC inhibitors in vivo and in vitro can inhibit tumor cell proliferation and induce tumor cell differentiation or apoptosis, a number of HDAC inhibitors such as LBH589 has entered Phase III clinical test, SAHA was approved in 2012 by the US FDA to market.

Another study showed that, in combination with anticancer drugs often show more than the original drug's anti-tumor effect, the cells from drying inhibitor chloroquine (Chloroquine, CQ), hydrogenated chloroquine (hydroxychloroquine, HCQ) and a number of cytotoxic Drug combinations have entered clinical trials, and the drug compound screening methods combined with SAHA joint programs with in vitro experiments also showed good results, but not yet.