WO2014203959A1 - PHLDA1又はPIK3C2Bの発現に基づくPI3K/AKT/mTOR阻害剤の治療効果の予測方法 - Google Patents
PHLDA1又はPIK3C2Bの発現に基づくPI3K/AKT/mTOR阻害剤の治療効果の予測方法 Download PDFInfo
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Definitions
- the present invention relates to a method, an antitumor agent, and a kit for predicting a therapeutic effect on chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor.
- Non-Patent Document 1 The PI3K / AKT / mTOR signal pathway controls various important cell functions such as cell proliferation, resistance to apoptosis, and glucose metabolism, but it is known that the activity of the signal pathway is enhanced in a wide range of malignant tumors.
- Non-Patent Document 1 Many clinical trials of PI3K / AKT / mTOR inhibitors (such as PI3K inhibitors, AKT inhibitors, mTOR inhibitors, or PI3K-mTOR dual inhibitors) are ongoing as antitumor agents. In clinical trials in which the targeted inhibitor is used alone, it has not been able to show a sufficient clinical effect (Non-patent Document 2).
- molecular target drugs such as PI3K / AKT / mTOR inhibitors may show strong efficacy against tumor cells in which the target molecule is highly expressed or highly active. It is important to stratify using predictive markers (Non-patent Document 3). Regarding PI3K / AKT / mTOR inhibitors, PIK3CA mutation or PTEN deletion has been clinically verified as a drug efficacy prediction marker (Non-patent Document 4).
- An object of the present invention is to provide a chemotherapy using a PI3K / AKT / mTOR inhibitor that exhibits an excellent antitumor effect on cancer patients.
- PI3K / AKT / mTOR inhibitors especially trans-3-amino-1-methyl-3- (4- (3-phenyl-5H-imidazo [1,2-c] pyrido [3,4-e] [1 , 3] oxazin-2-yl) phenyl) cyclobutanol) has been found to exhibit a significantly strong anti-tumor effect, leading to the completion of the present invention.
- the present invention provides the following method, antitumor agent and kit.
- [1] Based on the expression level of PHLDA1 and / or PIK3C2B in tumor cells of cancer patients, the therapeutic effect of chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patients is predicted Method.
- [2] The method according to [1], comprising the following steps (1) and (2).
- a PI3K / AKT / mTOR inhibitor is represented by the general formula (I)
- R 1a , R 1b , R 1c and R 1d each represent a hydrogen atom, and the other is a halogen atom, a cyano group, or a C1-6 alkyl group optionally having a hydroxyl group as a substituent, C1-6 An alkoxy group, a hydroxyl group as a substituent, an amino group, an optionally substituted mono or di (C1-6 alkyl) amino group or a carbonyl group having a mono or di (C1-6 alkoxy) amino group Represents an unsaturated heterocyclic group, R 2 represents a phenyl group, a pyridyl group, or a thienyl group, R 3 represents a hydrogen atom,
- A, B, C, and D represent CR 1a , CR 1b , CR 1c , and CR 1d , respectively. Or any one or two of A, B, C, and D is substituted with an N atom, At least two of R 1a , R 1b , R 1c and R 1d represent a hydrogen atom, and the other represents a chlorine atom, a fluorine atom, a cyano group, a methyl group, a hydroxymethyl group, a methoxy group, an ethoxy group, a carboxyl group, and a carbamoyl group, respectively.
- An antitumor agent containing a PI3K / AKT / mTOR inhibitor can exhibit a sufficient therapeutic effect by a method comprising the following steps (1) and (2) An antitumor agent characterized by being administered to a cancer patient who is predicted to be highly prone.
- a step of predicting that there is a high possibility that chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patient has a sufficient therapeutic effect.
- PHLDA1 for predicting the therapeutic effect of chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in cancer patients by a method comprising the following steps (1) and (2): / Or kit containing a reagent for measuring the expression level of PIK3C2B: (1) a step of measuring the expression level of PHLDA1 and / or PIK3C2B in a biological sample containing tumor cells collected from the patient, and (2) the expression level of PHLDA1 obtained in the above step (1) is preset.
- the anti-cancer agent containing a PI3K / AKT / mTOR inhibitor in the cancer patient A step of predicting that chemotherapy using a tumor agent is likely to have a sufficient therapeutic effect.
- chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor is likely to have a sufficient therapeutic effect by the method according to any one of [1] to [6] A method for treating a cancer patient, comprising administering the antitumor agent to the treated cancer patient.
- the present invention further includes the following aspects.
- [12] Based on the expression level of PHLDA1 and / or PIK3C2B in tumor cells of cancer patients, the therapeutic effect of chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patients is examined.
- Method. [13] The method according to [12], comprising the following steps (1) and (2).
- a step of determining that chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patient is highly likely to exhibit a sufficient therapeutic effect.
- the prediction method of the present invention can predict a cancer patient remarkably sensitive to a PI3K / AKT / mTOR inhibitor based on the expression of PHLDA1 and / or PIK3C2B. This makes it possible to select an appropriate drug in cancer treatment or avoid unnecessary medication, and to make an appropriate administration plan or change to an appropriate administration plan.
- the correlation of the expression level of PHLDA1 or PIK3C2B and the sensitivity with respect to a PI3K / AKT / mTOR inhibitor is shown.
- the analysis results of PHLDA1 gene knockdown, protein expression level and phosphorylation state are shown.
- the analysis result of the sensitivity change with respect to PI3K / AKT / mTOR inhibitor by PIK3C2B gene knockdown is shown.
- a comparison between the conventional method of measuring PIK3CA mutation / PTEN deletion and the measurement of expression level of PHLDA1 / PIK3C2B in predicting the therapeutic effect of PI3K / AKT / mTOR inhibitor is shown.
- the prediction method of the present invention is based on the expression level of PHLDA1 and / or PIK3C2B in a biological sample containing tumor cells collected from a cancer patient, and contains an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patient. This predicts or examines the therapeutic effect of chemotherapy using.
- cancers subject to the present invention include head and neck cancer, digestive organ cancer (esophageal cancer, gastric cancer, duodenal cancer, liver cancer, biliary tract cancer (eg, gallbladder / bile duct cancer), spleen cancer, and small intestine cancer.
- digestive organ cancer esophageal cancer, gastric cancer, duodenal cancer, liver cancer, biliary tract cancer (eg, gallbladder / bile duct cancer), spleen cancer, and small intestine cancer.
- Colorectal cancer colorectal cancer, colon cancer, rectal cancer, etc.
- lung cancer non-small cell lung cancer, small cell lung cancer, etc.
- breast cancer ovarian cancer
- uterine cancer cervical cancer, endometrial cancer, etc.
- kidney Cancer successful cancer removal, prostate cancer and the like can be mentioned.
- the cancer includes not only the primary lesion but also cancer that has metastasized to other organs (eg, liver).
- cancer patients include not only patients who actually have tumor cells but also patients whose tumor cells have disappeared or cannot be confirmed by surgery or chemotherapy.
- the “PI3K / AKT / mTOR inhibitor” in the present invention is not particularly limited as long as it is an agent that has an activity of inhibiting signal transduction enhancement in the PI3K / AKT / mTOR signal pathway, and is a group consisting of PI3K, AKT and mTOR.
- inhibitors for one or more target molecules selected from A PI3K inhibitor, an AKT inhibitor, an mTOR inhibitor, or a PI3K-mTOR dual inhibitor is preferable. These inhibitors may have inhibitory activity against target molecules other than PI3K, AKT and mTOR.
- the “inhibitor” includes not only an agent that inhibits the activity of the target molecule but also an agent that inhibits the expression of the target molecule. Moreover, there is no restriction
- a specific PI3K / AKT / mTOR inhibitor includes an imidoxoxazine compound represented by the general formula (I) or a salt thereof.
- the imidazooxazine compound represented by the general formula (I) is useful as an enzyme inhibitor for AKT.
- A, B, C, and D represent CR 1a , CR 1b , CR 1c , and CR 1d , respectively, or A, B, C, It shows that any one or two of D are substituted with N atoms.
- Examples of the halogen atom represented by R 1a , R 1b , R 1c , or R 1d include a chlorine atom, a bromine atom, a fluorine atom, and an iodine atom, and preferably a chlorine atom or a fluorine atom.
- the C1-6 alkyl group of the “C1-6 alkyl group optionally having a hydroxyl group as a substituent” represented by R 1a , R 1b , R 1c , or R 1d is a straight chain having 1 to 6 carbon atoms. Or a branched alkyl group, such as methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, sec-butyl group, tert-butyl group, pentyl group, hexyl group, etc. And preferably a C1-3 alkyl group, more preferably a methyl group.
- the number of hydroxyl groups (substituents) is 0 to 2, preferably 0 or 1.
- C1-6 alkoxy group represented by R 1a , R 1b , R 1c , or R 1d represents a linear or branched alkoxy group having 1 to 6 carbon atoms, and includes a methoxy group, an ethoxy group, n-propoxy group, isopropoxy group, n-butoxy group, isobutoxy group, tert-butoxy group and the like are shown, preferably a C1-3 alkoxy group, more preferably a methoxy group or an ethoxy group.
- R 1a , R 1b , R 1c , or R 1d “a hydroxyl group, an amino group, an optionally substituted mono- or di (C 1-6 alkyl) amino group, or a mono- or di-substituent”
- a carbonyl group having any of (C1-6 alkoxy) amino groups means a carbonyl group having a hydroxyl group as a substituent (ie, a carboxyl group), a carbonyl group having an amino group as a substituent (ie, a carbamoyl group), a substituent A mono- or di (C1-6 alkyl) aminocarbonyl group, which may have a mono- or di (C1-6 alkoxy) aminocarbonyl group.
- R 1a , R 1b , R 1c , or R 1d “a hydroxyl group, an amino group, an optionally substituted mono- or di (C 1-6 alkyl) amino group, or a mono- or di-substituent”
- the carbonyl group represents an aminocarbonyl group having one or two C1-6 alkyl groups described above, preferably a mono- or di (C1-3 alkyl) aminocarbonyl group, more preferably a methylaminocarbonyl group, dimethylamino A carbonyl group and an ethylaminocarbonyl group.
- the substituent is preferably a hydroxyl group. When it has a substituent, the number of substituents is preferably one.
- the mono- or di (C1-6 alkoxy) aminocarbonyl group represents an aminocarbonyl group having one or two C1-6 alkoxy groups, preferably a mono- or di (C1-3 alkoxy) aminocarbonyl group. More preferably, it is an ethoxyaminocarbonyl group.
- R 1a , R 1b , R 1c , or R 1d “a hydroxyl group, an amino group, an optionally substituted mono- or di (C 1-6 alkyl) amino group, or a mono- or di-substituent”
- Particularly preferred as the “carbonyl group having any one of (C 1-6 alkoxy) amino group” is a carboxyl group, a carbamoyl group, a methylaminocarbonyl group, an ethylaminocarbonyl group, a hydroxyethylaminocarbonyl group, and an ethoxyaminocarbonyl group.
- the “unsaturated heterocyclic group” represented by R 1a , R 1b , R 1c , or R 1d is a monocyclic or bicyclic group having 1 to 4 heteroatoms of N, S, or O
- a 5- to 10-membered unsaturated heterocyclic group such as imidazolyl group, thienyl group, furyl group, pyrrolyl group, oxazolyl group, isoxazolyl group, thiazolyl group, isothiazolyl group, pyrazolyl group, triazolyl group, tetrazolyl group, pyridyl group , Pyrazyl group, pyrimidinyl group, pyridazinyl group, indolyl group, isoindolyl group, indazolyl group, benzofuranyl group, benzoimidazolyl group, benzoxazolyl group, benzothiazolyl group, purinyl group, quinolyl group, is
- imidazooxazine compound represented by the general formula (I) may be the compounds described in any of the following (a) to (t).
- Acid formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, citric acid, tartaric acid, carbonic acid, picric acid, methanesulfonic acid, paratoluene Acid addition salts with organic acids such as sulfonic acid and glutamic acid, inorganic bases such as sodium, potassium, magnesium, calcium and aluminum, organic bases such as methylamine, ethylamine, meglumine and ethanolamine, or lysine, arginine and ornithine And salts with basic amino acids and ammonium salts.
- the imidazooxazine compound represented by the general formula (I) includes optical isomers and hydrates.
- imidazooxazine compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof can be produced according to the method described in WO2012 / 137870.
- specific PI3K / AKT / mTOR inhibitors include AMG-319, AZD-6482, BYL-719, Copanlist (BAY-80-6946), GDC- 0032, GDC-0084, GSK-1059615, GSK-2126458, GSK-2636771, Idelalisib (CAL-101), IPI-145, MLN-1117 (INK-1117), PA-799 (CH-5132799), Pictilib (GDC) -0941), Polararisib (XL-147), SF-1126, Sonolisib (PX-866), Voxtalisib (SAR-245409, XL-765) and the like are AKT inhibitors.
- Afulerative hydrochloride (GSK-2110183), ARQ-092, AZD5363, Enzastaurin hydrochloride, GDC-0068, GSK-2141795, GSK690693, LY-2780301, PerQ As inhibitors, AZD-2014, AZD-8055, CC-115, CC-223, DS-3078, Everolimus, Temirolimus, ME-344, MLN-0128 (INK-128), OSI-027, PWT-33597, Ridaforolimus, Sirolimus, etc.
- Dactolisib as Al inhibitors BEZ235
- DS-7423 GDC-0980
- NVP-BGT-226, PF-04691502, PF-05212384 PWT-33 597.
- PWT-33 597 Dactolisib as Al inhibitors
- PI3K / AKT / mTOR inhibitors MK-2206, BEZ235, GDC-0941, Sirolimus, or trans-3-amino-1-methyl-3 from the viewpoint of the effect of stratification by PHLDA1 and / or PIK3C2B -(4- (3-phenyl-5H-imidazo [1,2-c] pyrido [3,4-e] [1,3] oxazin-2-yl) phenyl) cyclobutanol is more preferred, trans-3- Amino-1-methyl-3- (4- (3-phenyl-5H-imidazo [1,2-c] pyrido [3,4-e] [1,3] oxazin-2-yl) phenyl) cyclobutanol Particularly preferred.
- the dosage form of the antitumor agent containing the PI3K / AKT / mTOR inhibitor in the present invention is appropriately selected from generally known dosage forms according to the kind of PI3K / AKT / mTOR inhibitor, specifically Examples include oral preparations (tablets, coated tablets, powders, granules, capsules, liquids, etc.), injections, suppositories, patches, ointments and the like.
- the antitumor agent can be prepared by a generally known method using a pharmacologically acceptable carrier according to the administration form.
- Such carriers include various types commonly used for ordinary drugs, such as excipients, binders, disintegrants, lubricants, diluents, solubilizers, suspending agents, isotonic agents, pH.
- examples include regulators, buffers, stabilizers, colorants, flavoring agents, and flavoring agents.
- chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor means a chemotherapy in which an antitumor agent containing at least the PI3K / AKT / mTOR inhibitor of the present invention is administered.
- chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor alone chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in combination with another antitumor agent Is also included.
- the amount of the PI3K / AKT / mTOR inhibitor contained in the antitumor agent is not constant depending on the symptom of the patient to which the antitumor agent is to be applied, or the dosage form thereof. 0.05 to 1,000 mg, about 0.01 to 500 mg for injections, and about 1 to 1,000 mg for suppositories are desirable.
- the daily dose of the drug having the above dosage form varies depending on the patient's symptom, body weight, age, sex, etc., and cannot be determined unconditionally.
- the dose may be 5,000 mg, preferably 0.1 to 1,000 mg, and is preferably administered once a day or divided into 2 to 3 times a day.
- the administration schedule of the chemotherapy is appropriately selected depending on conditions such as the type of PI3K / AKT / mTOR inhibitor, presence or absence of concomitant drugs, pretreatment calendar, stage, presence or absence of metastasis, patient age, and sex.
- the chemotherapy may be performed with or without surgery to remove the tumor during or before or after the implementation period. Even if it is chemotherapy that does not involve removal of the tumor mainly for the purpose of prolonging the life, it is a neoadjuvant chemotherapy that removes the tumor that has become smaller after the chemotherapy mainly for the purpose of reducing the tumor. Alternatively, it may be a postoperative adjuvant chemotherapy in which chemotherapy is performed prophylactically after removal of a tumor mainly for the purpose of suppressing recurrence / metastasis.
- “therapeutic effect” can be evaluated by a tumor shrinking effect, a recurrence / metastasis inhibiting effect, a life prolonging effect and the like as described above.
- “Chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor in the cancer patient exhibits a sufficient therapeutic effect” means that the therapeutic effect in a PHLDA1 low expression patient or a PIK3C2B high expression patient is PHLDA1. It means that it is remarkably superior with a statistically significant difference compared to the therapeutic effect in patients with high expression or patients with low expression of PIK3C2B.
- the biological sample in the present invention is not particularly limited as long as it is a biological sample collected from a cancer patient and containing tumor cells.
- Body fluid blood, urine, hair, etc.
- tissue biological biopsy sample, extracted organ, etc.
- extracts and cultures thereof can be suitably selected according to the kind of biological sample.
- PHLDA1 (Peckstrin homology-like domain family A member 1), which is an index in the present invention, is a kind of nuclear protein having a pleckstrin homologous domain and has been reported to be involved in the anti-apoptotic action of insulin-like growth factors. . In addition, involvement in the PI3K / AKT / mTOR pathway is not known.
- PIK3C2B phosphophatidylinitol-4-phosphate 3-kinase, catalytic subtype type 2 beta
- PIK3C2B phosphophatidylinitol-4-phosphate 3-kinase, catalytic subtype type 2 beta
- the measurement target of the expression level is not particularly limited as long as the expression level can be measured quantitatively or semi-quantitatively, and the expression level of mRNA, the copy number of DNA, the expression level of protein, and the like. Can be mentioned.
- Northern blotting method When measuring mRNA expression level, Northern blotting method, RT-PCR method, real-time PCR method, DNA microarray method, in situ high using probe or primer that specifically hybridizes with mRNA of PHLDA1 or PIK3C2B It can be measured by a commonly used method for measuring the expression level of mRNA, such as a hybridization method.
- the usual DNA copy such as in situ hybridization, array CGH, DNA microarray, Southern blotting, etc. It can be measured by a number measurement method.
- the primer and probe in the present invention are known base sequence information of human PHLDA1 or human PIK3C2B DNA or mRNA (human PHLDA1 DNA: GenBankID NC_000012.11. MRNA: GenBankID NM_007350; human PIK3C2B DNA: GenBankID NC_000001.10, mRNA: Based on GenBankID NM_002646), a polynucleotide that specifically hybridizes with DNA or mRNA of human PHLDA1 or human PIK3C2B is prepared by a generally known method.
- the primer has a base length of 10 to 50 bases, preferably 15 to 50 bases, more preferably 18 to 35 bases.
- the base length of the primer is 15 base length to total base length, preferably 20 to total base length, more preferably 30 to total base length.
- the primer and probe need not be completely complementary as long as they specifically hybridize with the DNA or mRNA of PHLDA1 or PIK3C2B.
- Such primers and probes have a polymorphism of 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 95% or more, particularly preferably 98% or more compared to the corresponding base sequence. It is a nucleotide.
- “specifically hybridizes” means that a specific hybrid is formed and a nonspecific hybrid is not formed under stringent hybridization conditions.
- Stringent hybridization conditions can be determined based on the melting temperature (Tm) of a nucleic acid that forms a hybrid in accordance with a generally known method. As washing conditions capable of maintaining a specific hybridized state, the conditions are usually about “1 ⁇ SSC, 0.1% SDS, 37 ° C.”, more strictly “0.5 ⁇ SSC, 0.1% SDS, 42 ° C. ", And more strictly,” 0.1 ⁇ SSC, 0.1% SDS, 65 ° C ".
- the probe or primer may be labeled with a commonly used radioactive substance, fluorescent substance, chemiluminescent substance, or enzyme so that it can be easily detected.
- the antibody in the present invention is not particularly limited as long as it specifically recognizes PHLDA1 or PIK3C2B protein, and may be either a monoclonal antibody or a polyclonal antibody, such as Fab fragment or F (ab ′) 2 fragment.
- the antibody fragment may also be used.
- the antibody can be produced according to a commonly known method based on the known amino acid sequence information of human PHLDA1 or human PIK3C2B protein (human PHLDA1 GenBankID NP_031376; human PIK3C2B GenBankID NP_002637) (for example, current protocol) (Molecular Biology edit. Ausubel et al. (1987), Publ. John Wiley and Sons. Section 11.12-11.13).
- Abcam cat. # Ab67849, Abnova cat. # H00022822-B01P and anti-human PIK3C2B antibody include Abcam cat. # Ab 55589, Agent cat. # AP3309a can be used.
- the cancer patient when the expression level of PHLDA1 in tumor cells of a cancer patient is low and / or when the expression level of PIK3C2B is high, the cancer patient contains a PI3K / AKT / mTOR inhibitor. It is predicted that there is a high possibility of showing a sufficient therapeutic effect for chemotherapy using an antitumor agent.
- the expression level of PHLDA1 is low refers to the case where the expression level of PHLDA1 of the cancer patient is relatively low compared to the expression level of PHLDA1 in the entire cancer patient, and more specifically, In this case, the expression level of PHLDA1 is equal to or lower than a preset cutoff point.
- the cut-off point varies depending on various conditions such as the type of measurement object and the method of measurement. Therefore, in the present invention, an arbitrary cut-off point that can vary depending on these various conditions is used. It encompasses the invention broadly and is not limited to a particular value.
- a specific cut-off point can be obtained by various statistical analysis methods from the expression level of PHLDA1 in a cancer patient measured in advance.
- the average or median expression level of PHLDA1 in cancer patients a cutoff point that separates the low expression group and high expression group of PHLDA1, and the therapeutic effect (survival period of the low expression group and high expression group of PHLDA1) Etc.) and a value that makes the P value minimum and a value that makes the P value less than a certain level (for example, a value that makes the P value less than 0.1, a value that makes the P value less than 0.05)
- a certain level for example, a value that makes the P value less than 0.1, a value that makes the P value less than 0.05
- the average value and the median value of the expression level of PHLDA1 in cancer patients are preferable, and the average value of the expression level of PHLDA1 in cancer patients is more preferable.
- the expression level of PIK3C2B is high refers to a case where the expression level of PIK3C2B of the cancer patient is relatively high compared to the expression level of PIK3C2B in the entire cancer patient.
- the expression level of PIK3C2B is equal to or higher than a preset cutoff point.
- the cut-off point can be obtained in the same manner as the cut-off point in the expression level of PHLDA1, but the average or median expression level of PIK3C2B in cancer patients is preferable, and the expression level of PIK3C2B in cancer patients The average value of is more preferable.
- the present invention also provides cancer patients that are predicted or determined to be highly likely that chemotherapy using an antitumor agent containing a PI3K / AKT / mTOR inhibitor has a sufficient therapeutic effect by the prediction method of the present invention. Also provided are antitumor agents containing PI3K / AKT / mTOR inhibitors for treatment.
- the present invention also provides such a usage form of the antitumor agent of the present invention.
- the antitumor agent of the present invention may include an instruction manual, a procedure manual, etc. describing a procedure for carrying out the prediction method of the present invention.
- the present invention also provides a kit for predicting or examining the therapeutic effect of chemotherapy using the antitumor agent of the present invention in cancer patients.
- the kit of the present invention is suitably used for predicting or examining the therapeutic effect of chemotherapy by the prediction method of the present invention described above.
- the kit of the present invention is specific to the DNA or mRNA of human PHLDA1 or human PIK3C2B. Primers and / or probes that are hybridizing polynucleotides may be included.
- the kit of the present invention comprises an antibody against PHLDA1 and / or PIK3C2B (enzyme),
- a secondary antibody against the antibody can be included, and the secondary antibody is preferably labeled with a luciferase label, a radioactive label, a fluorescent label, an enzyme label, or the like.
- kit of the present invention may include an instruction manual and a procedure manual describing procedures for carrying out the prediction method of the present invention.
- Example 1 (Correlation between expression level of PHLDA1 or PIK3C2B and sensitivity to PI3K / AKT / mTOR inhibitor) Human breast cancer-derived 37 cell lines (AU565, BT20, BT474, BT549, CAMA-1, DU4475, HCC1187, HCC1419, HCC1428, HCC1500, HCC1569, HCC1599, HCC1806, HCC1937, HCC1954, HCC202, HCC2218, HCC38, MBC70MD -157, MDA-MB-175, MDA-MB-231, MDA-MB-361, MDA-MB-415, MDA-MB-436, MDA-MB-453, MDA-MB-468, UACC-812, UACC -893 (above, obtained from American Type Culture Collection (ATCC)), MCF-7
- PI3K / AKT / mTOR inhibitor serially diluted with dimethyl sulfoxide (AKT inhibitor: Compound-I, MK-2206, PI3K-mTOR dual inhibitor: BEZ235, PI3K inhibitor: GDC-0941, mTOR inhibitor: Sirolimus) And only paclitaxel or dimethyl sulfoxide was added to the medium of each cell line.
- the amount of luminescence derived from living cells in the well at the time of drug addition was measured with a microplate reader (PerkinElmer, ARVOsx).
- the cell line to which only the drug or dimethyl sulfoxide was added was further cultured for 3 days in an incubator containing 37% at 5% carbon dioxide gas. After culturing, the mixture was allowed to stand at room temperature for 30 minutes, and 100 ⁇ L of the supernatant was removed from each well so that 100 ⁇ L of cell culture solution remained. An equivalent amount of CellTiter-Glo Luminescent Cell Viability Assay was added to 100 ⁇ L of the remaining cell culture solution.
- the amount of luminescence derived from living cells in each well was measured with a microplate reader.
- the cell growth rate is calculated from the following formula, and the concentration at which the cell growth rate is 50%, that is, the concentration of each drug that inhibits cell growth by 50% (GI 50 value ( ⁇ M)). Asked.
- the extracted total RNA was checked for concentration and purity using a spectrophotometer (Nanodrop 1000, Thermo Fisher Scientific). Moreover, the decomposition degree was confirmed using Agilent 2100 Bioanalyzer (Agilent Technologies). Using GeneChip 3 ′ IVT Express Kit (Affymetrix), the extracted total RNA was reverse transcribed into cDNA and further subjected to a transcription reaction to biotinylated cRNA.
- the amplified biotinylated cRNA (20 ⁇ g) was then fragmented. Each operation was performed according to the description in the instructions attached to the kit. Using the obtained biotinylated cRNA derived from each cell line as a specimen, GeneChip Human Genome U-133 Plus 2.0 Array (Affymetrix, PIK3C2B probe set ID: 204484_at, PHLDA1 probe set ID: 2179p_atb) It moved to 640 (Affymetrix) and hybridization was performed under the conditions of 45 ° C. and 60 rpm for 16 hours.
- GeneChip Human Genome U-133 Plus 2.0 Array Affymetrix, PIK3C2B probe set ID: 204484_at, PHLDA1 probe set ID: 2179p_atb
- FIG. 1 shows the Pearson correlation between the logarithm of the GI 50 value of each drug and the logarithm of the expression levels of PHLDA1 and PIK3C2B obtained as described above.
- a 240 cell line panel composed of various cancer types such as lung cancer, kidney cancer, liver cancer, pancreatic cancer, colon cancer, bladder cancer, breast cancer, head and neck cancer, prostate cancer, skin cancer, blood cancer, malignant lymphoma, etc.
- OncoPanel TM Ricerca Biosciences
- PHLDA1 or PIK3C2B correlates with the sensitivity to PI3K / AKT / mTOR inhibitors.
- the GI 50 values of a wide range of PI3K / AKT / mTOR inhibitors (Compound-I, MK-2206, BEZ235, GDC-0941, Sirolimus) It was confirmed that the logarithm and the logarithm of the expression level of PHLDA1 and PIK3C2B were significantly correlated.
- Example 2 (PHLDA1 gene knockdown, protein expression level and phosphorylation status analysis) The relationship between PHLDA1 and the PI3K / AKT / mTOR signal pathway was verified by confirming the phosphorylation state of AKT when the PHLDA1 gene was knocked down.
- HCC38, HCC1806, HCC1937, all obtained from American Type Culture Collection (ATCC)) in which PHLDA1 was relatively highly expressed were RPMI medium (Life Technologies Corp., Cat #) containing 10% FBS. A104910-01).
- siRNA ON-TARGET Plus control pool, Thermo Fisher Scientific, Cat # D-001810-10-20-20
- PHLDA1-siRNA ON-TARGET Plus siRNA HumanCHF # J-012389-08
- siRNA was introduced into cells using Lipofectamine RNAiMAX (Life Technologies, Cat # 13778-150), and gene knockdown was performed.
- each cell line was washed twice with cold PBS (Life Technologies, Cat # 10010-023), solubilized buffer (PhosSTOP (Roche, Cat # 4906837) and Complete, EDTA-free (Roche, Cat) Pierce RIPA Buffer (Thermo Fisher Scientific, Cat # 89901)) to which # 1873580) was added and solubilized by cooling with ice for 30 minutes.
- the solubilized extract was centrifuged at 15000 rpm for 30 minutes at 4 ° C., and the supernatant was recovered to obtain a protein extract.
- the obtained protein extract was measured for protein concentration according to the manual using BCA Protein Assay Reagent (Thermo Fisher Scientific, Cat # 23225).
- the protein extract was denatured by adding a reducing buffer (ImmunoPure Lane Maker Reduce Sample Buffer (Thermo Fisher Scientific, Cat # 39000)) and boiling. SDS-PAGE was performed by applying 20 ⁇ g of the reduced protein extract per lane to Criterion TGX Precast Gels (Biorad, Cat # 567-1084).
- a reducing buffer ImmunoPure Lane Maker Reduce Sample Buffer (Thermo Fisher Scientific, Cat # 39000)
- SDS-PAGE was performed by applying 20 ⁇ g of the reduced protein extract per lane to Criterion TGX Precast Gels (Biorad, Cat # 567-1084).
- the sample was transferred to a PVDF membrane (Trans-Blot Turbo Midi PVDF Transfer Pack, Biorad, Cat # 170-4157) using a blotting apparatus (Trans-Blot Turbo Transfer System, Biorad).
- the transfer membrane was blocked by blocking with Blocking One-P (Nacalai, Cat # 05999-84) at room temperature for 1 hour.
- Blocking One-P Nacalai, Cat # 05999-814
- anti-human PHLDA1 goat antibody (Santacruz, Cat # sc-6142) diluted 1000-fold with TBS buffer (hereinafter abbreviated as TBS-T) supplemented with 5% BSA and 0.05% Tween 20, The reaction was carried out overnight at 4 ° C.
- PVDF membrane was washed 3 times with TBS-T at room temperature for 10 minutes, and then diluted with 2000% diluted 5% skim milk / TBS-T (horseradish peroxidase (HRP) labeled anti-goat antibody (Santacruz, Cat # Sc-2020) and an anti-rabbit antibody (GE Healthcare, Cat # NA9340V)) at room temperature for 1 hour.
- HRP horseradish peroxidase
- GE Healthcare Cat # NA9340V
- the PVDF membrane was washed three times with TBS-T at room temperature for 10 minutes, and then chemiluminescence was detected using ECL Prime Western Blotting Detection Reagent (GE Healthcare, Cat # RPN2232) using LAS-3000 (FujiFilm). . The result is shown in FIG.
- PHLDA1 gene knockdown resulted in increased AKT phosphorylation (S473), confirming activation of PI3K / AKT / mTOR signal pathway. It was done. Since the phosphorylation (S473) level of AKT correlates with the in vitro and in vivo anti-tumor effects on PI3K inhibitors (Cancer Research 70, 4982-4994 (2010)), the expression level of PHLDA1 is PI3K / AKT / It has been shown that it can be involved in therapeutic effects on mTOR inhibitors.
- Human ovarian cancer cell line OVCAR-3 (obtained from ATCC) is RPMI medium (Life Technologies Corp., Cat # A104910-) containing 20% FBS, 0.01 mg / mL bovine insulin (Sigma-Aldrich, Cat # 10516). 01).
- Human breast cancer cell line HCC1187 (obtained from ATCC) was cultured in RPMI medium (Life Technologies Corp., Cat # A104910-01) containing 10% FBS.
- each cell line was used to perform cytotoxicity tests against PI3K / AKT / mTOR inhibitor (AKT inhibitor: Compound-I, PI3K inhibitor: GDC-0941) and paclitaxel.
- FIG. 3 shows the results of measurement of the relative number of cells after treatment with each concentration of drug, with the number of cells of control (dimethylsulfoxide added) cultured for 3 days being 100 and no cell being 0.
- Example 4 Comparison of conventional method of measuring PIK3CA mutation / PTEN deletion and measurement of expression level of PHLDA1 / PIK3C2B in predicting the therapeutic effect of PI3K / AKT / mTOR inhibitor.
- the PIK3CA mutation or PTEN deletion which has been verified as a drug efficacy prediction marker in the clinic, and the expression level of PHLDA1 or PIK3C2B were compared from the viewpoint of the correlation with the sensitivity to the PI3K / AKT / mTOR inhibitor.
- FIG. 4 shows the GI 50 value ( ⁇ M) for Compound-I depending on the presence or absence of PIK3CA mutation (E545K or H1047R) or PTEN deletion in these 36 strains.
- PTEN deletion (8 strains) could not be correlated with sensitivity to Compound-I.
- the relationship between the expression level of PHLDA1 or PIK3C2B and the sensitivity to Compound-I was verified.
- FIG. 4 shows the distribution of GI 50 values ( ⁇ M) for Compound-I when the average expression level of PHLDA1 or PIK3C2B is divided into two groups using the cut-off point as an average value.
- the expression level of PHLDA1 or PIK3C2B clearly has a higher correlation with sensitivity to Compound-I than the conventional method of PIK3CA mutation or PTEN deletion, and is useful as a clinical efficacy prediction marker.
Abstract
Description
本願は、2013年6月20日に出願した特願2013-129591号明細書(その全体が参照により本明細書中に援用される)の優先権の利益を主張するものである。
(技術分野)
本発明は、PI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法に対する治療効果を予測する方法、抗腫瘍剤及びキットに関する。
〔1〕 がん患者の腫瘍細胞におけるPHLDA1及び/又はPIK3C2Bの発現量に基づき、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法の治療効果を予測する方法。
〔2〕 下記工程(1)及び(2)を含む、〔1〕記載の方法。
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下である場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上である場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。
〔3〕 PI3K/AKT/mTOR阻害剤が、一般式(I)
R1a、R1b、R1c、R1dのうち少なくとも2つが水素原子を示し、他方がそれぞれ
ハロゲン原子、シアノ基、置換基としてヒドロキシル基を有してもよいC1-6アルキル基、C1-6アルコキシ基、置換基としてヒドロキシル基、アミノ基、置換基を有してもよいモノ又はジ(C1-6アルキル)アミノ基又はモノ又はジ(C1-6アルコキシ)アミノ基のいずれかを有するカルボニル基、不飽和複素環基を示し、
R2は、フェニル基、ピリジル基、又はチエニル基を示し、
R3は、水素原子、メチル基、エチル基、又はシクロプロピル基を示し、
R4は、水素原子、又はヒドロキシ基を示す。)
で表されるイミダゾオキサジン化合物又はその薬学的に許容される塩であるか、
AMG-319、AZD-6482、BYL-719、Copanlisib(BAY-80-6946)、GDC-0032、GDC-0084、GSK-1059615 、GSK-2126458、GSK-2636771、Idelalisib(CAL-101)、IPI-145、MLN-1117(INK-1117)、PA-799(CH-5132799)、Pictilisib(GDC-0941)、Pilaralisib(XL-147)、SF-1126、Sonolisib(PX-866) 、Voxtalisib(SAR-245409、XL-765)、Afuresertib hydrochloride(GSK-2110183 )、ARQ-092、AZD5363、Enzastaurin hydrochloride、GDC-0068、GSK-2141795、GSK690693、LY-2780301、MK-2206、Perifosine、Triciribine phosphate(VQD-002)、AZD-2014、AZD-8055、CC-115、CC-223、DS-3078、Everolimus、Temsirolimus、ME-344、MLN-0128(INK-128)、OSI-027、PWT-33597、Ridaforolimus、Sirolimus、Dactolisib(BEZ235) 、DS-7423、GDC-0980、NVP-BGT-226、PF-04691502、PF-05212384(PKI-587)、又はPWT-33597である〔1〕又は〔2〕記載の方法。
〔4〕 一般式(I)で表されるイミダゾオキサジン化合物が、A、B、C、及びDは、それぞれC-R1a、C-R1b、C-R1c、及びC-R1dを示すか、或いは前記A、B、C、Dのうちいずれか1又は2つがN原子に置換されていることを示し、
R1a、R1b、R1c、R1dのうち少なくとも2つが水素原子を示し、他方がそれぞれ塩素原子、フッ素原子、シアノ基、メチル基、ヒドロキシメチル基、メトキシ基、エトキシ基、カルボキシル基、カルバモイル基、メチルアミノカルボニル基、エチルアミノカルボニル基、ヒドロキシエチルアミノカルボニル基、エトキシアミノカルボニル基、ピラゾリル基を示し、
R2は、フェニル基、ピリジル基、又はチエニル基を示し、
R3は、水素原子、メチル基、エチル基、又はシクロプロピル基を示し、
R4は、水素原子、又はヒドロキシ基を示す化合物である、〔3〕記載の方法。
〔5〕 一般式(I)で表されるイミダゾオキサジン化合物が、次の(a)~(t)のいずれかの化合物である〔3〕又は〔4〕記載の方法。
(b)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-フルオロ-3-(ピリジン-4-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(c)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(d)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(e)トランス-3-アミノ-1-シクロプロピル-3-(4-(9-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(f)トランス-3-アミノ-1-シクロプロピル-3-(4-(8-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(g)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(h)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(i)トランス-3-アミノ-1-エチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(j)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(k)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e] [1,3]オキサジン-2-イル)フェニル)シクロブタノール
(l)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(m)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(n)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,2-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(o)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピラジノゾ[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(p)トランス-3-アミノ-3-(4-(9-(ヒドロキシメチル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)-1-メチルシクロブタノール
(q)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-9-カルボニトリル
(r)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-9-(1H-ピラゾール-5-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(s)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-メチル-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド
(t)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-エトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド
〔6〕 PI3K/AKT/mTOR阻害剤が、MK-2206、BEZ235、GDC-0941、Sirolimus、トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e] [1,3]オキサジン-2-イル)フェニル)シクロブタノールである〔1〕~〔5〕のいずれか1項記載の方法。
〔7〕 腫瘍細胞におけるPHLDA1の発現量が予め設定されたカットオフポイント以下であるか、又は腫瘍細胞におけるPIK3C2Bの発現量が予め設定されたカットオフポイント以上であるがん患者を治療するためのPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤。
〔8〕 PI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤であって、下記工程(1)及び(2)を含む方法により当該抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者に投与されることを特徴とする抗腫瘍剤。
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下の場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上の場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。
〔9〕 下記工程(1)及び(2)を含む方法により、がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法の治療効果を予測するための、PHLDA1及び/又はPIK3C2Bの発現量を測定する試薬を含有するキット:
(1)当該患者から採取された腫瘍細胞を含む生体試料におけるPHLDA1及び/又はPIK3C2Bの発現量を測定する工程、及び
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下の場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上の場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。
〔10〕 〔1〕~〔6〕のいずれか1項記載の方法によりPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者に、該抗腫瘍剤を投与することを含むがん患者の治療方法。
〔11〕 〔1〕~〔6〕のいずれか1項記載の方法によりPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者の治療に用いるための、PI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤。
〔12〕 がん患者の腫瘍細胞におけるPHLDA1及び/又はPIK3C2Bの発現量に基づき、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法の治療効果を検査する方法。
〔13〕下記工程(1)及び(2)を含む、〔12〕記載の方法。
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下の場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上の場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと判定する工程。
本発明における「PI3K/AKT/mTOR阻害剤」とは、PI3K/AKT/mTORシグナルパスウェイにおけるシグナル伝達の亢進を阻害する活性を有する薬剤であれば特に制限されず、PI3K、AKT及びmTORからなる群から選ばれる1あるいは2以上の標的分子に対する阻害剤が挙げられる。好ましくはPI3K阻害剤、AKT阻害剤、mTOR阻害剤又はPI3K-mTORデュアル阻害剤である。なお、これら阻害剤は、PI3K、AKT及びmTOR以外の標的分子に対する阻害活性を併せ持っていても良い。ここで「阻害剤」には、標的分子の活性を阻害する薬剤のみならず、標的分子の発現を阻害する薬剤をも包含する。また、PI3K/AKT/mTOR阻害剤の態様としては、特に制限はなく、低分子化合物、抗体、アンチセンスオリゴヌクレオチド、siRNA及びアプタマー等が挙げられる。
(b)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-フルオロ-3-(ピリジン-4-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(c)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(d)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(e)トランス-3-アミノ-1-シクロプロピル-3-(4-(9-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(f)トランス-3-アミノ-1-シクロプロピル-3-(4-(8-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(g)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(h)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(i)トランス-3-アミノ-1-エチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(j)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(k)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e] [1,3]オキサジン-2-イル)フェニル)シクロブタノール(以下、compound-Iと称す。)
(l)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(m)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール (n)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,2-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(o)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピラジノゾ[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(p)トランス-3-アミノ-3-(4-(9-(ヒドロキシメチル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)-1-メチルシクロブタノール
(q)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-9-カルボニトリル
(r)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-9-(1H-ピラゾール-5-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(s)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-メチル-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド
(t)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-エトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド
一般式(I)で表されるイミダゾオキサジン化合物の薬学的に許容される塩としては、塩酸、臭化水素酸、ヨウ化水素酸、硫酸、硝酸、リン酸等の無機酸や、ギ酸、酢酸、プロピオン酸、シュウ酸、マロン酸、コハク酸、フマル酸、マレイン酸、乳酸、リンゴ酸、クエン酸、酒石酸、炭酸、ピクリン酸、メタンスルホン酸、パラトルエンスルホン酸、グルタミン酸などの有機酸との酸付加塩、ナトリウム、カリウム、マグネシウム、カルシウム、アルミニウムなどの無機塩基や、メチルアミン、エチルアミン、メグルミン、エタノールアミンなどの有機塩基、又はリジン、アルギニン、オルニチンなどの塩基性アミノ酸との塩やアンモニウム塩が挙げられる。また、一般式(I)で表されるイミダゾオキサジン化合物には、光学異性体も含まれ、水和物も含まれる。
抗腫瘍剤中に含有されるPI3K/AKT/mTOR阻害剤の量は、これを適用すべき患者の症状により、或いはその剤形等により一定ではないが、一般に投与単位形態あたり、経口剤では約0.05~1,000mg、注射剤では約0.01~500mg、坐剤では約1~1,000mgとするのが望ましい。また、上記投与形態を有する薬剤の1日あたりの投与量は、患者の症状、体重、年齢、性別等によって異なり一概には決定できないが、通常成人(体重50kg)1日あたり約0.05~5,000mg、好ましくは0.1~1,000mgとすればよく、これを1日1回又は2~3回程度に分けて投与するのが好ましい。
[実施例1]
(PHLDA1又はPIK3C2Bの発現量と、PI3K/AKT/mTOR阻害剤に対する感受性との相関)
ヒト乳癌由来37細胞株(AU565、BT20、BT474、BT549、CAMA-1、DU4475、HCC1187、HCC1419、HCC1428、HCC1500、HCC1569、HCC1599、HCC1806、HCC1937、HCC1954、HCC202、HCC2218、HCC38、HCC70、MDA-MB-157、MDA-MB-175、MDA-MB-231、MDA-MB-361、MDA-MB-415、MDA-MB-436、MDA-MB-453、MDA-MB-468、UACC-812、UACC-893(以上、American Type Culture Collection(ATCC)より入手)、MCF-7、SK-BR-3、T47D、ZR-75-1(以上、大日本住友製薬株式会社より入手)、KPL-1、KPL-3C、KPL-4(紅林博士(川崎医大)より供与))を用いて以下のとおり細胞毒性試験を行った。各細胞株は96ウェル平底マイクロプレートに150μL播種し、37℃、5%炭酸ガス含有の培養器中で1日培養した。ジメチルスルホキシドにて段階稀釈したPI3K/AKT/mTOR阻害剤(AKT阻害剤:Compound-I、MK-2206、PI3K-mTORデュアル阻害剤:BEZ235、PI3K阻害剤:GDC-0941、mTOR阻害剤:Sirolimus)及びパクリタキセル、あるいはジメチルスルホキシドのみを各細胞株の培地に添加した。これを先に述べた96ウェル平底マイクロプレートの各ウェルに50μL加えて、薬剤の最終濃度がそれぞれ10000、3000、1000、300、100、30、3、1nMになるようにした。また、別途用意した、各細胞株を一日間培養した96ウェル平底マイクロプレートを室温に30分間放置後、各ウェルから上清を50μLずつ除き、100μLの細胞培養液が残るようにした。残った細胞培養液100μLに対し、等量のCellTiter-Glo Luminescent Cell Viability Assay(Promega、Cat#:G7573)を各ウェルに添加した。10分間暗所で放置した後、マイクロプレートリーダー(PerkinElmer、ARVOsx)にて薬剤添加時のウェルの生細胞由来発光量を測定した。薬剤あるいはジメチルスルホキシドのみを加えた細胞株は37℃、5%炭酸ガス含有の培養器中でさら3日間培養した。培養後、室温に30分間放置し、各ウェルから上清を100μLずつ除き、100μLの細胞培養液が残るようにした。残った細胞培養液100μLに対し、等量のCellTiter-Glo Luminescent Cell Viability Assayを添加した。10分間暗所で放置した後、マイクロプレートリーダーにて各ウェルの生細胞由来発光量を測定した。ヒト乳癌由来37細胞株において、以下の式より細胞増殖率を算出し、細胞増殖率が50%となる濃度、すなわち細胞増殖を50%阻害する各薬剤の濃度(GI50値(μM))を求めた。
細胞増殖率(%)=(T-C0)/C0×100;T<C0の場合
C0:薬剤添加時のウェルの発光量(count per second)
C:ジメチルスルホキシドのみを添加したウェルの発光量(count per second)
T:被検薬剤を添加したウェルの発光量(count per second)
次いで、ヒト乳癌由来37細胞株におけるPHLDA1及びPIK3C2Bの発現量を測定した。各細胞株は、10cm細胞培養ディッシュで50~80%コンフルエント程度になるまで培養後、RNeasy Mini Kit(QIAGEN、Cat#74106)を用いてマニュアルに従いtotal RNAを抽出した。抽出したtotal RNAは、分光光度計(Nanodrop 1000、Thermo Fisher Scientific)を用いて濃度及び純度を確認した。また、Agilent 2100 Bioanalyzer(Agilent Technologies)を用いて分解度を確認した。GeneChip 3' IVT Express Kit(Affymetrix)を用いて、抽出したtotal RNAをcDNAに逆転写し、さらにビオチン化cRNAへの転写反応を行った。次いで、増幅したビオチン化cRNA(20μg)を断片化した。なお各操作は、キットに添付の説明書の記載に従って行った。得られた各細胞株由来のビオチン化cRNAを検体として、GeneChip Hunman Genome U-133 Plus 2.0 Array(Affymetrix、PIK3C2B probe set ID:204484_at、PHLDA1 probe set ID:217997_at)に添加し、GeneChip Hybridization Oven 640(Affymetrix)に移して、45℃、60rpmの条件下で16時間ハイブリダイゼーションを行った。ハイブリダイゼーション後、GeneChipFluidic Station 450(Affymetrix)を用いて洗浄および蛍光標識を行い、GeneChip Scanner 3000 7G(Affymetrix)を用いてスキャンしてマイクロアレイ・データを取得した。マイクロアレイ・データの解析には、MATLAB(MathWorks)を使用した。遺伝子発現量は、RMA法(Robust Multiarray Average法)により計算した。
(PHLDA1遺伝子ノックダウン、タンパク質の発現量及びリン酸化状態の解析)
PHLDA1遺伝子をノックダウンした場合における、AKTのリン酸化状態を確認することにより、PHLDA1とPI3K/AKT/mTORシグナルパスウェイの関係を検証した。
(PIK3C2B遺伝子ノックダウンによるPI3K/AKT/mTOR阻害剤に対する感受性変化の解析)
ヒト卵巣癌細胞株OVCAR-3(ATCCより入手)は、20% FBS、0.01 mg/mL bovine insulin(Sigma-Aldrich、Cat#I0516)を含むRPMI培地(Life Technologies Corp. 、Cat# A104910-01 )で培養した。ヒト乳癌細胞株HCC1187(ATCCより入手)は、10% FBSを含むRPMI培地(Life Technologies Corp.,Cat# A104910-01)で培養した。
(PI3K/AKT/mTOR阻害剤治療効果予測性における、従来法PIK3CA変異・PTEN欠失測定とPHLDA1・PIK3C2B発現量測定との比較)
臨床において薬効予測マーカーとして検証されているPIK3CA変異あるいはPTEN欠失と、PHLDA1あるいはPIK3C2B発現量について、PI3K/AKT/mTOR阻害剤に対する感受性との相関性の観点から比較した。
Claims (11)
- がん患者から採取した腫瘍細胞におけるPHLDA1及び/又はPIK3C2Bの発現量に基づき、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法の治療効果を予測する方法。
- 下記工程(1)及び(2)を含む、請求項1記載の方法:
(1)当該患者から採取された腫瘍細胞を含む生体試料におけるPHLDA1及び/又はPIK3C2Bの発現量を測定する工程、及び
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下である場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上である場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。 - PI3K/AKT/mTOR阻害剤が、一般式(I)
R1a、R1b、R1c、R1dのうち少なくとも2つが水素原子を示し、他方がそれぞれ
ハロゲン原子、シアノ基、置換基としてヒドロキシル基を有してもよいC1-6アルキル基、C1-6アルコキシ基、置換基としてヒドロキシル基、アミノ基、置換基を有してもよいモノ又はジ(C1-6アルキル)アミノ基又はモノ又はジ(C1-6アルコキシ)アミノ基のいずれかを有するカルボニル基、不飽和複素環基を示し、
R2は、フェニル基、ピリジル基、又はチエニル基を示し、
R3は、水素原子、メチル基、エチル基、又はシクロプロピル基を示し、
R4は、水素原子、又はヒドロキシ基を示す。)
で表されるイミダゾオキサジン化合物又はその薬学的に許容される塩であるか、
AMG-319、AZD-6482、BYL-719、Copanlisib(BAY-80-6946)、GDC-0032、GDC-0084、GSK-1059615 、GSK-2126458、GSK-2636771、Idelalisib(CAL-101)、IPI-145、MLN-1117(INK-1117)、PA-799(CH-5132799)、Pictilisib(GDC-0941)、Pilaralisib(XL-147)、SF-1126、Sonolisib(PX-866) 、Voxtalisib(SAR-245409、XL-765)、Afuresertib hydrochloride(GSK-2110183 )、ARQ-092、AZD5363、Enzastaurin hydrochloride、GDC-0068、GSK-2141795、GSK690693、LY-2780301、MK-2206、Perifosine、Triciribine phosphate(VQD-002)、AZD-2014、AZD-8055、CC-115、CC-223、DS-3078、Everolimus、Temsirolimus、ME-344、MLN-0128(INK-128)、OSI-027、PWT-33597、Ridaforolimus、Sirolimus、Dactolisib(BEZ235) 、DS-7423、GDC-0980、NVP-BGT-226、PF-04691502、PF-05212384(PKI-587)、又はPWT-33597である請求項1又は2記載の方法。 - 一般式(I)で表されるイミダゾオキサジン化合物が、A、B、C、及びDは、それぞれC-R1a、C-R1b、C-R1c、及びC-R1dを示すか、或いは前記A、B、C、Dのうちいずれか1又は2つがN原子に置換されていることを示し、
R1a、R1b、R1c、R1dのうち少なくとも2つが水素原子を示し、他方がそれぞれ塩素原子、フッ素原子、シアノ基、メチル基、ヒドロキシメチル基、メトキシ基、エトキシ基、カルボキシル基、カルバモイル基、メチルアミノカルボニル基、エチルアミノカルボニル基、ヒドロキシエチルアミノカルボニル基、エトキシアミノカルボニル基、ピラゾリル基を示し、
R2は、フェニル基、ピリジル基、又はチエニル基を示し、
R3は、水素原子、メチル基、エチル基、又はシクロプロピル基を示し、
R4は、水素原子、又はヒドロキシ基を示す化合物である、請求項3記載の方法。 - 一般式(I)で表されるイミダゾオキサジン化合物が、次の(a)~(t)のいずれかの化合物である請求項3又は4記載の方法。
(a) トランス-3-アミノ-1-シクロプロピル-3-(4-(10-フルオロ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(b)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-フルオロ-3-(ピリジン-4-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(c)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(d)トランス-3-アミノ-1-シクロプロピル-3-(4-(10-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(e)トランス-3-アミノ-1-シクロプロピル-3-(4-(9-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(f)トランス-3-アミノ-1-シクロプロピル-3-(4-(8-メトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(g)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(h)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(i)トランス-3-アミノ-1-エチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(j)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(k)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e] [1,3]オキサジン-2-イル)フェニル)シクロブタノール
(l)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(m)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[4,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(n)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,2-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(o)トランス-3-アミノ-1-シクロプロピル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピラジノゾ[2,3-e][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(p)トランス-3-アミノ-3-(4-(9-(ヒドロキシメチル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)-1-メチルシクロブタノール
(q)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-9-カルボニトリル
(r)トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-9-(1H-ピラゾール-5-イル)-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-2-イル)フェニル)シクロブタノール
(s)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-メチル-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド
(t)2-(4-(トランス-1-アミノ-3-ヒドロキシ-3-メチルシクロブチル)フェニル)-N-エトキシ-3-フェニル-5H-ベンゾ[e]イミダゾ[1,2-c][1,3]オキサジン-8-カルボキサミド - PI3K/AKT/mTOR阻害剤が、MK-2206、BEZ235、GDC-0941、Sirolimus、トランス-3-アミノ-1-メチル-3-(4-(3-フェニル-5H-イミダゾ[1,2-c]ピリド[3,4-e] [1,3]オキサジン-2-イル)フェニル)シクロブタノールである請求項1~5のいずれか1項記載の方法。
- 腫瘍細胞におけるPHLDA1の発現量が予め設定されたカットオフポイント以下であるか、又は腫瘍細胞におけるPIK3C2Bの発現量が予め設定されたカットオフポイント以上であるがん患者を治療するためのPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤。
- PI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤であって、下記工程(1)及び(2)を含む方法により当該抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者に投与されることを特徴とする抗腫瘍剤:
(1)当該患者から採取された腫瘍細胞を含む生体試料におけるPHLDA1及び/又はPIK3C2Bの発現量を測定する工程、及び
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下の場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上の場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。 - 下記工程(1)及び(2)を含む方法により、がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法の治療効果を予測するための、PHLDA1及び/又はPIK3C2Bの発現量を測定する試薬を含有するキット:
(1)当該患者から採取された腫瘍細胞を含む生体試料におけるPHLDA1及び/又はPIK3C2Bの発現量を測定する工程、及び
(2)上記工程(1)で得られたPHLDA1の発現量が予め設定されたカットオフポイント以下の場合、又は上記工程(1)で得られたPIK3C2Bの発現量が予め設定されたカットオフポイント以上の場合、当該がん患者におけるPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測する工程。 - 請求項1~6のいずれか1項記載の方法によりPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者に、該抗腫瘍剤を投与することを含むがん患者の治療方法。
- 請求項1~6のいずれか1項記載の方法によりPI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤を用いた化学療法が十分な治療効果を示す可能性が高いと予測されたがん患者の治療に用いるための、PI3K/AKT/mTOR阻害剤を含有する抗腫瘍剤。
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WO2017134030A1 (en) * | 2016-02-01 | 2017-08-10 | Bayer Pharma Aktiengesellschaft | Copanlisib biomarkers |
WO2017134000A1 (en) * | 2016-02-01 | 2017-08-10 | Bayer Pharma Aktiengesellschaft | Copanlisib biomarkers |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
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Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010087497A1 (ja) * | 2009-02-02 | 2010-08-05 | 財団法人ヒューマンサイエンス振興財団 | 医薬組成物、および、腫瘍の治療用医薬剤 |
WO2012137870A1 (ja) | 2011-04-06 | 2012-10-11 | 大鵬薬品工業株式会社 | 新規イミダゾオキサジン化合物又はその塩 |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8445198B2 (en) * | 2005-12-01 | 2013-05-21 | Medical Prognosis Institute | Methods, kits and devices for identifying biomarkers of treatment response and use thereof to predict treatment efficacy |
EP2452281B1 (en) * | 2009-07-08 | 2019-10-02 | Worldwide Innovative Network | Method for predicting efficacy of drugs in a patient |
CN104066735B (zh) | 2012-01-10 | 2016-08-31 | 拜耳知识产权有限责任公司 | 作为akt激酶抑制剂的取代的咪唑并吡嗪 |
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Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2010087497A1 (ja) * | 2009-02-02 | 2010-08-05 | 財団法人ヒューマンサイエンス振興財団 | 医薬組成物、および、腫瘍の治療用医薬剤 |
WO2012137870A1 (ja) | 2011-04-06 | 2012-10-11 | 大鵬薬品工業株式会社 | 新規イミダゾオキサジン化合物又はその塩 |
Non-Patent Citations (13)
Title |
---|
ARCARO A. ET AL.: "Two distinct phosphoinositide 3-kinases mediate polypeptide growth factor-stimulated PKB activation", THE EMBO JOURNAL, vol. 21, no. 19, 2002, pages 5097 - 5108, XP055301014 * |
AUSUBEL ET AL.: "Current Protocols in Molecular Biology", 1987, PUBLISH. JOHN WILEY AND SONS |
CANCER RESEARCH, vol. 70, 2010, pages 4982 - 4994 |
DOMIN J. ET AL.: "The class II phosphoinositide 3-kinase PI3K-C2beta regulates cell migration by a PtdIns(3)P dependent mechanism", JOURNAL OF CELLULAR PHYSIOLOGY, vol. 205, no. 3, 2005, pages 452 - 462, XP055301019 * |
LANCET ONCOLOGY, vol. 12, 2011, pages 594 - 603 |
LIU P. ET AL.: "Targeting the phosphoinositide 3-kinase (PI3K) pathway in cancer", NATURE REVIEWS DRUG DISCOVERY, vol. 8, no. 8, 2009, pages 627 - 644, XP055020926 * |
LIU Z. ET AL.: "Phosphatidylinositol 3-kinase-C2? inhibits cisplatin-mediated apoptosis via the Akt pathway in oesophageal squamous cell carcinoma", THE JOURNAL OF INTERNATIONAL MEDICAL RESEARCH, vol. 39, no. 4, 2011, pages 1319 - 1332, XP055301016 * |
NATURE REVIEWS CANCER, vol. 2, 2002, pages 489 - 501 |
NATURE REVIEWS CANCER, vol. 9, 2009, pages 550 - 562 |
NATURE REVIEWS CLINICAL ONCOLOGY, vol. 8, 2011, pages 587 - 596 |
See also references of EP3012327A4 |
TRAER C.J. ET AL.: "Are class II phosphoinositide 3-kinases potential targets for anticancer therapies?", BULLETIN DU CANCER, vol. 93, no. 5, 2006, pages 53 - 58, XP002697164 * |
WANG X. ET AL.: "PHLDA1/2 contribute to tumor suppression in breast and lung cancer as downstream targets of oncogenic HER2 signaling", CANCER RESEARCH, vol. 72, no. ISSUE, 2012, PROCEEDINGS OF THE 103RD ANNUAL MEETING OF THE AACR 2012;CHICAGO, XP008182592 * |
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Publication number | Publication date |
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PL3543355T3 (pl) | 2021-05-31 |
JPWO2014203959A1 (ja) | 2017-02-23 |
HUE053611T2 (hu) | 2021-07-28 |
PT3543355T (pt) | 2021-02-19 |
KR20180069141A (ko) | 2018-06-22 |
DK3543355T3 (da) | 2021-03-01 |
DK3012327T3 (da) | 2021-01-25 |
KR102279579B1 (ko) | 2021-07-21 |
RU2666921C2 (ru) | 2018-09-13 |
AU2014282179A1 (en) | 2016-01-28 |
US10155990B2 (en) | 2018-12-18 |
PL3012327T3 (pl) | 2021-04-19 |
US20160102366A1 (en) | 2016-04-14 |
JP6289459B2 (ja) | 2018-03-07 |
HUE053322T2 (hu) | 2021-06-28 |
ES2844200T3 (es) | 2021-07-21 |
TWI626443B (zh) | 2018-06-11 |
EP3012327A4 (en) | 2017-03-22 |
ES2855698T3 (es) | 2021-09-24 |
PT3012327T (pt) | 2021-01-22 |
RU2016101364A (ru) | 2017-07-26 |
EP3012327B1 (en) | 2020-12-16 |
TW201819923A (zh) | 2018-06-01 |
TW201537174A (zh) | 2015-10-01 |
EP3543355A1 (en) | 2019-09-25 |
EP3012327A1 (en) | 2016-04-27 |
AU2014282179B2 (en) | 2017-09-14 |
EP3543355B1 (en) | 2020-12-23 |
KR20160021836A (ko) | 2016-02-26 |
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