KR101971236B1 - Marker for predicting susceptibility to combination therapy - Google Patents

Abstract

The present invention provides a marker composition and kit for predicting susceptibility of cancer cells to a combination of PI3K inhibitor and Src inhibitor, comprising an agent for measuring the phosphorylation level of phosphatidylinositol-3-kinases (PI3K) subunit p85, phosphorylation of p85 in biological samples. Measuring the level; It relates to a method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a. A marker composition comprising an agent for measuring the phosphorylation level of the phosphatidylinositol-3-kinases (PI3K) subunit p85 according to the present invention and a method using the same are used to determine whether to co-administer a PI3K inhibitor and a Src inhibitor to a cancer patient. Cancer patients who may be effective by the co-administration of drugs can be classified quickly and early, and the patient-specific chemotherapy can be efficiently performed.

Description

Marker for predicting susceptibility to combination therapy}

The present invention provides a marker composition and kit for predicting the susceptibility of cancer cells to a combination of PI3K inhibitor and Src inhibitor, comprising an agent for measuring the phosphorylation level of phosphatidylinositol-3-kinases (PI3K) subunit p85, phosphorylation of p85 in biological samples Measuring the level; It relates to a method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a.

Even in today's rapidly evolving medical technologies, cancer is still one of the most threatening diseases of humanity, and many people are diagnosed with cancer and die from cancer or its complications. According to the prevalence rate, the seven major cancers in Korea are gastric cancer, liver cancer, lung cancer, colon cancer, uterine cancer, breast cancer, and head and neck cancer. Two lung cancers and head and neck cancer belonging to upper digestive system cancer are known to be correlated. Upper aerodigestive tract (UADT) cancer can be interpreted as a generic term for tumors that occur in various parts of the body, including the larynx, oral cavity, oropharyngeal, nasal, sinus, nasopharynx, hypopharyngeal, and salivary glands.

The incidence of cancers in the upper gastrointestinal respiratory system, such as head and neck cancers, is increasing gradually, and the limit of continuous resistance to cisplatin despite the use of cisplatin, the most powerful anticancer drug currently used in the clinic for the treatment of cancer. There is this. Therefore, there is a great need for a new anticancer agent that can replace it, and recently, a study for developing a new anticancer agent targeting Phosphatidylinositol 3-kinase (PI3K) / AKT / mTOR signaling system has been conducted.

The PI3K / AKT / mTOR signaling system is an important factor that regulates basic cellular functions such as cancer cell metabolism, growth, survival, and angiogenesis. In general, activation of the PI3K mechanism is responsible for cancer cell progression, metastasis, poor prognostic factors, and radiation. It is known as a factor indicating resistance to treatment. Therefore, the development of drugs that inhibit the PI3K / Akt / mTOR pathway, various drugs are currently in clinical trials. In particular, the development of drugs that target PI3K, which plays a key role in the pathway, can effectively inhibit PI3K-addicted cancer, and these PI3K inhibitors are expected to play a role in increasing the sensitivity of cancer cells. However, in some clinical applications, resistance to PI3K inhibitors has been found in targeted therapies with PI3K inhibitors, indicating technical limitations in the treatment of cancer. Nevertheless, there has not been much research on the exact mechanism of resistance and resistance to PI3K inhibitors and the application of new therapies. Therefore, there is a need for a new method to solve the problem of cancer cell resistance to PI3K inhibitors. There is this.

Therefore, the present inventors have studied the mechanism of resistance to PI3K inhibitors and the appropriate treatment method to solve the resistance, it was confirmed that the mutual regulation of phosphorylation and Src phosphorylation of p85, a subunit of PI3K is related to PI3K inhibitor resistance. The present invention was completed by confirming that phosphorylation of PI3K subunit p85 can be used as a biomarker for determining co-administration of PI3K inhibitor and Src inhibitor.

Accordingly, an object of the present invention is to provide a marker composition and kit for predicting the sensitivity of cancer cells to co-administration of PI3K inhibitor and Src inhibitor, comprising an agent for measuring phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85.

It is also an object of the present invention to measure the phosphorylation level of p85 in a biological sample; To provide a method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a.

It is also an object of the present invention to provide a pharmaceutical composition for preventing or treating PI3K inhibitor resistant cancer, comprising a PI3K inhibitor and a Src inhibitor.

It is also an object of the present invention to process the candidate substance in PI3K inhibitor resistant cancer cells; Selecting a substance that inhibits PI3K p85 phosphorylation and Src phosphorylation; It provides a method for screening a PI3K inhibitor resistance inhibitory substance comprising a.

In order to achieve the above object, the present invention provides a marker composition for predicting the sensitivity of cancer cells to co-administration of PI3K inhibitor and Src inhibitor, comprising an agent for measuring phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85. .

The present invention also provides a kit for predicting susceptibility of cancer cells to a combination of PI3K inhibitor and Src inhibitor, comprising an agent for measuring phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85.

In another aspect, the present invention comprises the steps of measuring the phosphorylation level of p85 in the biological sample; It provides a method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a.

The present invention also provides a pharmaceutical composition for preventing or treating PI3K inhibitor resistant cancer, comprising a PI3K inhibitor and a Src inhibitor.

The present invention also provides a method of treating a PI3K inhibitor resistant cancer cell with a candidate substance; Selecting a substance that inhibits PI3K p85 phosphorylation and Src phosphorylation; It provides a method for screening a PI3K inhibitor resistance inhibitory substance comprising a.

A marker composition comprising an agent for measuring the phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85 according to the present invention and a method using the same are used to determine whether to co-administer a PI3K inhibitor and an Src inhibitor to a cancer patient. It is possible to quickly classify cancer patients that may be effective by the combined administration, so that patient-specific chemotherapy can be efficiently performed.

1 is a diagram showing the results of confirming whether phosphorylation of subunit p85 of PI3K in AMC-HN-3, HN30, and HN31 cell lines.
Figure 2 is a diagram showing the results confirming the effect of cancer cell killing and the phosphorylation level of PI3K p85 according to the treatment of PI3K inhibitors (LY294002, BYL719) in AMC-HN-3, HN30, HN31 cell line (* p <0.05).
Figure 3 shows the results of comparing the degree of phosphorylation of Src in the AMC-HN-3, HN30, HN31 cell line (A) and Src phosphorylation inhibitory effect (B) according to the treatment of PI3K inhibitors (LY294002, BYL719) It is also.
Figure 4 shows the results confirming colony formation and reduction of phosphorylation of Src and p85 in the PI3K inhibitor resistant HN31 cell line, either alone or in combination with the Src inhibitor dasatinib and the PI3K inhibitor (LY294002, BYL719).
5 is a schematic diagram showing the mechanisms involved in p85 phosphorylation and Src phosphorylation interregulation in PI3K inhibitor sensitive cancer cells and PI3K inhibitor resistant cancer cells.

Hereinafter, the present invention will be described in more detail.

The present invention provides a marker composition for predicting the susceptibility of cancer cells to a combination of PI3K inhibitor and Src inhibitor, comprising an agent measuring the phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85.

Through the preparation for measuring phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85 according to the present invention, when phosphorylation of p85 is confirmed in cancer cells, the cancer cells may be predicted to be susceptible to co-administration of a PI3K inhibitor and a Src inhibitor. This can effectively select patients suitable for co-administration.

In addition, the marker composition may be characterized in that it further comprises an agent for measuring the phosphorylation level of Src.

In the present invention, the "agent for measuring phosphorylation level" may include without limitation agents used in conventional phosphorylation measurement methods known in the art, such as antibodies that recognize phosphoryl groups, chemicals that recognize phosphoryl groups, luminescence The agent used for the law etc. can be used. The chemical substance that recognizes the phosphoryl group may be a molecular probe such as a Zn2 + complex-based biotinylated phosphate-specific ligand based on Zn2 + complex, and specifically, may be a phos-tag. More preferably, the agent for measuring phosphorylation level of the present invention may be an antibody or phos-tag for measuring phosphorylation level.

Methods for determining the phosphorylation level using the agent for measuring the phosphorylation level are Western blot, MALDI-TOF / MS, Enzyme linked immunosorbent assay (ELISA), radioisotope labeling method and various known to those skilled in the art It is preferably selected from the group consisting of a labeling method and the like, but is not limited thereto. Specifically, Western blot and ELISA can be performed using a phosphorylated substrate-specific antibody, radioisotope labeling method has the ability to adsorb with [ ³² P] γ-ATP and biotinylated kinase substrate, avidin Phosphorylation can be measured using a membrane adsorbed. In addition, the phosphorylation activity can be measured using a moving car on the electrophoretic phase due to the net charge difference of the substrate after phosphorylation without using radioisotopes. Can be used.

Phosphorylation level measurement according to the present invention can be used as a meaning including both the presence and absence of phosphorylation and phosphorylation, in the determination of phosphorylation level to determine whether or not to administer the Src inhibitor in combination, the determination of the phosphorylation level of PI3K p85 May be to confirm the presence or absence of phosphorylation of p85, the measurement of the Src phosphorylation level may preferably be to confirm that the Src phosphorylation is increased compared to PI3K sensitive cells.

The cancer cells according to the present invention may be characterized as being PI3K inhibitor resistant cancer, and the term "resistance" means that there is no significant change in cell proliferation despite treatment with an anticancer agent. The PI3K inhibitor-resistant cancer according to the present invention may be characterized by exhibiting phosphorylation at PI3K p85 and maintaining resistance by activating Src phosphorylation and p85, thereby maintaining resistance.

On the other hand, in the present invention, "sensitization" refers to a state in which cell proliferation is effectively changed by an anticancer agent, that is, a state in which cancer cells are killed, and a state in which an anticancer effect desired by anticancer agent can be achieved. Therefore, in the present invention, the prediction of the sensitivity of cancer cells to the co-administration of PI3K inhibitor and Src inhibitor is to predict whether cancer cells, preferably PI3K inhibitor-resistant cancer cells can be effectively killed by co-administration of PI3K inhibitor and Src inhibitor. it means.

In the present invention, the cancer may be an upper gastrointestinal respiratory system cancer, and specific examples include head and neck cancer, laryngeal cancer, pharyngeal cancer, oral cancer, oropharyngeal cancer, nasopharyngeal cancer, nasal cancer, mesothelial cancer, lung cancer and esophageal cancer. It may be more than one cancer.

In the present invention, “PI3K inhibitor” is defined as LY294002 (CAS number: 447154447-36-6), BYL719 (CAS number: 1217486-61-7), Quercetin (CAS number: 117-39-5), Wortmannin (CAS number: 19545-26-7), Idelalisib (CAS number: 870281-82-6) and Pictilisib (CAS number: 957054-30-7) It may be one or more selected from the group consisting of, more preferably LY294002, BYL719 or not limited thereto.

Also in the present invention, "Src" refers to a protein corresponding to EC 2.7.10.2. Src is a non-receptor tyrosine kinase and primary cancer gene. As used herein, an "Src inhibitor" inhibits the production of the expression product of the Src gene (by stopping Src gene transcription and / or blocking the translation of mRNA from Src gene expression) and inhibiting Src protein activity. It refers to any molecule capable of completely or partially inhibiting Src gene expression by directly inhibiting it. As a specific example, the Src inhibitor of the present invention may be preferably at least one selected from the group consisting of Dasatinib, Bosutinib, and saracatinib, and most preferably Dasatinib. However, it is not limited thereto.

In another aspect, the present invention provides a kit for predicting the sensitivity of cancer cells to a combination of the PI3K inhibitor and the Src inhibitor comprising the predictive composition.

"Kit" according to the present invention is a combination of a PI3K inhibitor and an Src inhibitor by comprising a composition comprising an agent for measuring the phosphorylation level of phosphatidylinositol-3-kinases (PI3K) p85 and / or an agent for measuring the phosphorylation level of Src Means a tool that can predict the susceptibility of cancer cells to. In addition to the above composition, the kit of the present invention may include other constituent compositions, solutions, and devices necessary for predicting susceptibility, wherein the first compartment measures the phosphorylation level of PI3K p85, and the second compartment measures the phosphorylation level of Src. The agent may be divided and included.

In another aspect, the present invention provides a method of preparing a biocompatible product comprising the steps of: measuring the phosphorylation level of p85 in a biological sample; It provides a method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a; measuring the phosphorylation level of Src; It may further include.

In the present invention, the biological sample may be cancer tissue or cancer cells.

Specifically, the method may be performed by the following method.

Determining the phosphorylation level of p85 in the biological sample can be carried out through the agents and methods for measuring the phosphorylation level of PI3K p85 as described above, if the phosphorylation of p85 in the biological sample, preferably cancer cells is confirmed, Cancer cells and / or patients can be diagnosed as having cancer cells susceptible to co-administration of PI3K inhibitors and Src inhibitors. In addition, if the Src phosphorylation level is measured by an agent and method for measuring the phosphorylation level of Src, and the phosphorylation level is increased compared to control cells susceptible to PI3K inhibitors, the cancer cells and / or cancer patients may use a PI3K inhibitor and a Src inhibitor in combination. It can be diagnosed as being sensitive to administration.

Through the above diagnostic method, cancer patients who can overcome the resistance to the PI3K inhibitor and obtain an anticancer effect through the co-administration of the PI3K inhibitor and the Src inhibitor can be selected early, and patient-specific treatment can be performed.

The present invention also provides a pharmaceutical composition for preventing or treating PI3K inhibitor resistant cancer, comprising a PI3K inhibitor and a Src inhibitor.

Concomitant administration of PI3K inhibitors and Src inhibitors can relieve PI3K inhibitor resistance and induce effective killing of cancer cells. Particularly preferred subjects are those in which PI3K p85 phosphorylation and PI3K inhibitor sensitive cancer cells are identified. In comparison, cancers show a sustained increase in Src phosphorylation.

The PI3K inhibitor and the Src inhibitor are the same as described above, and in one specific example of the present invention, by combining LY294002 or BYL719 as a PI3K inhibitor and dasatinib as a Src inhibitor to PI3K inhibitor resistant cancer cells in which phosphorylation of p85 is confirmed, It was confirmed that phosphorylation of p85 was reduced and Src phosphorylation was effectively inhibited to induce death of cancer cells. Concomitant administration in the present invention is meant to include all dosage acts administered sequentially or simultaneously.

The composition of the present invention comprises a pharmaceutically acceptable carrier. Carriers included in the pharmaceutical composition of the present invention and acceptable carriers are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, Microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, saline, phosphate buffered saline (PBS) or And the like, but are not limited thereto.

In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical compositions of the present invention may be administered orally or parenterally, preferably orally or subcutaneously, and may be used together, simultaneously, and sequentially, including additional ingredients for anticancer treatment.

Suitable dosages of the pharmaceutical compositions of the present invention may be prescribed in various ways depending on factors such as formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to response of the patient. Can be.

The present invention may also include a kit or a commercial package comprising a PI3K inhibitor and a Src inhibitor for the treatment of a PI3K inhibitor resistant cancer, wherein the kit or the commercial package is a simultaneous treatment of a PI3K inhibitor and a Src inhibitor in the treatment of a PI3K inhibitor resistant cancer. It may contain instructions for red, separate or sequential use.

The invention also encompasses the use of Src inhibitors in combination with PI3K inhibitors in the manufacture of a medicament for use in the treatment of PI3K inhibitor resistant cancer.

The present invention also provides a method of treating a PI3K inhibitor resistant cancer cell with a candidate substance; Selecting a substance that inhibits PI3K p85 phosphorylation and Src phosphorylation; It provides a method for screening a PI3K inhibitor resistance inhibitory substance comprising a.

The PI3K inhibitor resistance is due to p85 phosphorylation and increased phosphorylation of Src, and sustained phosphorylation by their mutual regulation. In one specific embodiment of the present invention, the PI3K inhibitor and Src inhibitor are administered to the PI3K inhibitor through co-administration. When the resistance to the problem is resolved and susceptibility to the concomitant treatment, the cancer cells die, the phosphorylation of Src is reduced, it was confirmed that the phosphorylation of PI3K p85 is also reduced. Thus, when identifying substances that inhibit PI3K p85 phosphorylation and Src phosphorylation in PI3K inhibitor resistant cells, they can be screened and developed as materials that can be used as PI3K inhibitor resistance inhibitors, such PI3K inhibitor resistance inhibitors, such as PI3K inhibitors. And combination dose formulations of Src inhibitors are available as therapeutics for PI3K inhibitor resistant cancer.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are merely to illustrate the invention, but the content of the present invention is not limited by the following examples.

Example  One. Phosphatidyl Inositol Identification of p85 Phosphorylation in 3-kinase Inhibitor-resistant Cell Lines

Experiments to compare the degree of PI3K / Akt activation in phosphatidylinositol-3-kinase (hereafter PI3K) resistant head and neck cancer cell lines (HN31) and susceptible cell lines (HN30, AMC-HN-3) Was performed. Among the cell lines used in the experiment, AMC-HN-3 is a head and neck cancer cell line which is separated from the head and neck cancer patients of Asan Hospital, and HN30 and HN31 are Dr. Dr. of the University of Texas MD Anderson cancer center. It was provided by Myer. Specifically, the phosphorylation-expressing protein-specific antibody of the three cells, AMC-HN-3, HN30, HN31, PI3K p85 phosphorylation (p-PI3K p85), AKT phosphorylation (p-AKT), T-AKT activation degree Confirmed using Western blot. Beta actin was used as a control, and the results of confirming the degree of PI3K / Akt activation in the resistant and sensitive cells are shown in FIG. 1.

As shown in FIG. 1, it was confirmed that the activation of PI3K / Akt was increased in the resistant cell line HN31 as compared with the sensitive cell lines HN3 and HN30. In particular, the phosphorylation of p85, a subunit of PI3K, was not found in the sensitive cell lines HN3 and HN30. However, the phosphorylation of p85 subunit was important in resistance to PI3K inhibitors. It was confirmed.

To confirm the association between PI3K inhibitor resistance and p85 phosphorylation, we identified the PI3K inhibitor Ly294002 and BYL719 for the PI3K inhibitor-resistant head and neck cancer cell lines HN31 and susceptible cell lines HN30 and AMC-HN3 that have increased expression of PI3K / Akt. Were treated by concentrations, and cell viability and IC50 values of cancer cells were determined, and phosphorylation levels of PI3K p85 were measured while Ly294002 (25uM) and BYL719 (10uM) were treated for 0 to 6 hours, respectively, and the results are shown in FIG. 2. . Cell survival after drug treatment was confirmed using MTT (3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) dye.

As shown in FIG. 2, the HN31 cell line was shown to be resistant to PI3K inhibitor despite the increased activation of PI3K / Akt. Specifically, the IC50 value of HN31 was about three times higher for LY294002 and BYL719 compared to HN30, a sensitive cell line.

In order to determine the difference in sensitivity, activation of PI3K-related protein was examined using Western blot. Phosphorylation of p85 was not found in the sensitive cell lines HN3 and HN30, but distinct p85 phosphorylation was observed in the resistant cell line HN31. In addition, it was confirmed that phosphorylation was not reduced and sustained activation was maintained even by the treatment of LY294002 and BYL719 over time.

These results confirmed that the resistance to PI3K inhibitors in resistant cell lines with increased PI3K / Akt activation was because the phosphorylation of PI3K p85 was not inhibited and remained active. That is, it can be seen that the resistance of cancer cells to PI3K inhibitors is indicated by the phosphorylation of PI3K subunit p85.

Example  2. p85 phosphorylation and Src  Confirmation of Mutual Regulation of Kinase Phosphorylation

As confirmed in Example 1, it was confirmed that the PI3K inhibitor-resistant cell line did not inhibit the phosphorylation of PI3K p85 and maintained the activity according to the sustained phosphorylation of p85 when the PI3K inhibitor was treated. It was. Activation of p85 is expected to involve an activation mechanism through phosphorylation of a member of a superfamily of membrane-associated nonreceptor protein tyrosine kinases (Src), one of the cross-signals of the PI3K / Akt signaling system. Phosphorylation of Src at, HN30 and HN31 was analyzed using Western blot. In addition, each cell was treated with PI3K inhibitors LY294002 (25uM) and BYL719 (10uM) for 0 to 6 hours, and the results of comparing the degree of phosphorylation of Thr416 (Src activating portion) of Src in cancer cells are shown in FIG. 3.

As shown in FIG. 3, it was confirmed that phosphorylation of Src, in particular, Thr416, was significantly increased in HN31, a cell line resistant to Western blot PI3K inhibitors (A). These results indicate that the resistance to PI3K inhibitors and the phosphorylation at the Thr416 position of Src are related.

In addition, treatment of PI3K inhibitors to HN31 cell lines resistant to PI3K inhibitors confirmed the sustained phosphorylation of Src, that is, maintenance of activation. On the other hand, sensitive AMC-HN-3 and HN30 cell lines significantly inhibited Src phosphorylation when treated with PI3K inhibitors (B).

Through the above results, in the resistance of cancer cells to PI3K inhibitors, continuous phosphorylation by the interregulation of PI3K p85 and Src plays an important role, and the presence or absence of p85 phosphorylation and the increase or decrease of Src phosphorylation to the resistance to PI3K inhibitors. It has been confirmed that it can be a target of marker or resistance reduction. That is, according to the results confirmed in the present invention, since phosphorylation of p85 and phosphorylation of Src plays an important role in PI3K inhibitor resistance, by screening a material that inhibits it, it is possible to suppress the resistance of the PI3K inhibitor and enhance the sensitivity It can be seen that the material can be selected.

Example  3. Src  Bio for inhibitor combination therapy As a marker p85  Check utility

Through Example 2, it was confirmed that continuous phosphorylation by the mutual regulation of PI3K p85 and Src is a mechanism that causes the resistance to PI3K inhibitors, so that PI3K inhibitors and Src inhibitors are used in combination with PI3K inhibitor resistant cells whose phosphorylation of PI3K p85 is confirmed. When treated, it was confirmed whether the ability to overcome the resistance to the PI3K inhibitor and achieve cancer cell killing effect, and thus the change of p85, Src phosphorylation level. To this end, PI3K inhibitors LY294002 (25uM), BYL719 (10uM) and Dasatinib (200nM) as Src inhibitors were administered separately or together, and the phosphorylation of Src, the inhibition of PI3K p85 phosphorylation and the effects of anticancer drugs Colonization assay was performed. For colony formation analysis, 14 days after treatment with PI3K inhibitor and Src inhibitor, colonies were evaluated as colonies, stained with crystal violet, and observed under a microscope. It was. In addition, the changes in Src phosphorylation and PI3K p85 phosphorylation by the drug alone and in combination administration were confirmed by Western blot and the results are shown in FIG. 4.

As shown in Figure 4, 14 days after the drug treatment colony formation assay results in the group treated with only the PI3K inhibitor without dasatinib treatment did not show the effect of anticancer drugs due to PI3K resistance, but administered together with dasatinib and PI3K inhibitor In one group, colony formation was significantly reduced (p <0.0001), suggesting that co-administration of Src inhibitors and PI3K inhibitors could block the mechanism of resistance to PI3K inhibitors and thus restore sensitivity to PI3K inhibitors. It was. These results demonstrate once again that phosphorylation of Src and p85 is involved in resistance to PI3K inhibitors.

In addition, Western blot confirmed the change in phosphorylation of p85 and Src when co-administered with dasatinib and PI3K inhibitors. In the group treated with dasatinib, the phosphorylation of Src and P85 was continuously maintained, while the combination of dasatinib and PI3K inhibitors was used. In the administration group, the phosphorylation at the Thr416 position of Src was clearly lost, and the phosphorylation level of PI3K p85 was also markedly decreased. These results suggest that sustained phosphorylation by the interregulation of phosphorylation of p85 and phosphorylation of Src causes resistance to PI3K inhibitors, and that inhibition of their sustained phosphorylation can restore sensitivity to PI3K inhibitors. will be.

In other words, since phosphorylation of p85 and phosphorylation of Src cause continuous phosphorylation by mutual regulation, treatment with Src inhibitor in PI3K inhibitor-resistant cancer cells that maintains the phosphorylation of p85 continuously blocks the phosphorylation by mutual regulation and the PI3K inhibitor. Sensitivity can be enhanced, such combination treatments include: phosphorylation of p85 in PI3K inhibitor resistant cancer cells; Or phosphorylation of p85 and phosphorylation of Src; Can be determined by identifying with a biomarker. In other words, when the phosphorylation of p85 is confirmed in cancer cells, especially PI3K inhibitor resistant cancer cells, or when the phosphorylation of p85 and increased phosphorylation of Src is confirmed, the anticancer effect when the cells are co-administered with PI3K inhibitor and Src inhibitor It can be determined as a cancer cell that can represent. In addition, when a combination of PI3K inhibitor and Src inhibitor is administered to cancer cells in which PI3K p85 phosphorylation is confirmed, anti-cancer effects may be enhanced by increasing sensitivity in PI3K inhibitor resistant cancer.

The mechanism of resistance to PI3K inhibitors identified through the above examples is shown schematically in FIG. 5.

Claims (11)

A marker composition for predicting susceptibility of cancer cells to co-administration of a PI3K inhibitor and an Src inhibitor, comprising an agent measuring the phosphorylation level of the phosphatidylinositol-3-kinases (PI3K) subunit p85.
The marker composition for predicting susceptibility according to claim 1, further comprising an agent for measuring phosphorylation level of Src.
The marker composition for predicting susceptibility according to claim 1, wherein the cancer cells are PI3K inhibitor resistant cancer cells.
According to claim 1, The cancer cells are head and neck cancer, laryngeal cancer, pharyngeal cancer, oral cancer, oropharyngeal cancer, nasopharyngeal cancer, nasal cancer, mesothelial cancer, lung cancer and esophageal cancer, one or more cancer cells selected from the group consisting of markers for predicting susceptibility .
A kit for predicting susceptibility of cancer cells to a combination of a PI3K inhibitor and a Src inhibitor comprising the composition of any one of claims 1 to 4.
Measuring the phosphorylation level of p85 in the biological sample; Method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor comprising a.
The method of claim 6, further comprising: measuring phosphorylation level of Src; Method for providing information on the susceptibility diagnosis of cancer cells for co-administration of PI3K inhibitor and Src inhibitor further comprising.
delete delete delete Treating the candidate with PI3K inhibitor resistant cancer cells; And
Selecting a substance that inhibits phosphorylation and Src phosphorylation of PI3K subunit p85; Screening method of PI3K inhibitor resistance inhibitory substance comprising a.

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