RU2848373C1 - 2h-[1,4]oxazino- and 2h-[1,4]thiazino[3,2-c]quinoline-3(4h)-one derivatives as atm and dna-pk kinase inhibitors for the treatment and/or prevention of cancer - Google Patents

Abstract

FIELD: pharmaceuticals.

SUBSTANCE: invention relates to the use of a pharmaceutical composition having inhibitory activity against ATM and DNA-PK, containing a therapeutically effective amount of a compound defined by formula (I) or a pharmaceutically acceptable salt thereof, wherein Y may be -S- or -SO₂-, then Z₁ and Z₂ are independently selected and represent -CH- or N; or Y may be represent -O-, then either Z₁ or Z₂ represents N and the other represents -CH-; M₁ and M₂ represent hydrogen or M₁ and M₂ together form =O (keto group); n is 0 or 1; R₁ is -O-L-N(Ra,Rb); R₂ is C_1-3 alkyl, C₃ cycloalkyl or phenyl; R₃ is halogen; R₄ is hydrogen or C_1-2 alkyl; each R_a and R_b independently is hydrogen or C_1-3 alkyl; L is ethylene or n-propylene, and a pharmaceutically acceptable excipient for the manufacture of a medicament for the treatment of cancer treatable with a dual ATM and DNA-PK inhibitor, and the use of the composition for the manufacture of a medicament for the treatment of cancer treatable with a dual ATM and DNA-PK inhibitor, in a patient receiving an antineoplastic agent selected from the following list: cisplatin, oxaliplatin, carboplatin, valrubicin, idarubicin, calicheamicin, olaparib, rucaparib, niraparib, velaparib, talazoparib, ipilimumab, ofatumumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab.

EFFECT: new pharmaceutical composition that can be used to produce a drug for the treatment of cancer that is treatable with a dual ATM and DNA-PK inhibitor.

10 cl, 2 tbl

Description

The invention relates to the chemistry of heterocyclic organic compounds and their pharmaceutical compositions. These compounds and their pharmaceutical compositions selectively inhibit the activity of ataxia telangiectasia mutated ("ATM" - mutant protein in ataxia-telangiectasia) and DNA-dependent protein kinase ("DNA-PK" - DNA-dependent protein kinase) serine/threonine protein kinases. In this regard, the invention also relates to a method of using such compounds and their compositions for the treatment and/or prevention of oncological diseases as monotherapy and/or in combination with radiotherapy, chemotherapy and/or immunotherapy.

Members of the phosphatidylinositol 3-kinase-related kinase (PIKK) family—ATM and DNA-PK kinases—are important cellular modulators whose activity is particularly associated with oxidative stress and DNA damage, particularly double-strand breaks. Clinical trials have shown that inhibiting the activity of these kinases (either individually or simultaneously) using small molecule drugs significantly increases the sensitivity of tumor cells to ionizing radiation during radiotherapy and the action of certain classes of anticancer drugs. However, these molecules themselves have no effect on normal cells. Therefore, dual ATM and DNA-PK inhibitors in combination with radiotherapy, chemotherapy, and/or immunotherapy can be used for the effective treatment of cancer, significantly potentiating the effect of these therapeutic approaches. The use of such molecules allows, in particular, for lower doses of radiation or DNA-targeted antitumor drugs, thereby reducing unwanted side effects. Treatment of patients with dual ATM/DNA-PK inhibitors slows or stops DNA repair after radiotherapy. This significantly enhances apoptosis in tumor cells that survive therapeutic treatment. For example, mice with mutations in the ATM and PRKDC genes, which lead to loss of function of the ATM and DNA-PK proteins they encode, respectively, exhibited hypersensitivity to ionizing radiation. Simultaneous inhibition of ATM and DNA-PK is expected to more effectively sensitize tumor cells to radiation or other DNA-damaging agents compared to inhibition of either kinase alone. Minimizing off-target inhibition of related kinases, such as mTOR or PI3Ks, may reduce the toxicity of this class of inhibitors. Within the framework of the present invention, dual inhibitors of ATM and DNA-PK kinases were developed that do not exhibit pharmaceutically significant inhibitory activity against related kinases (PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ, and mTOR). Such molecules are capable of significantly increasing the sensitivity of tumor cells to radiation and/or certain chemotherapeutic agents.

Currently, several small molecule drugs are in early-stage clinical trials as monotherapies (MT), chemo- (CS), and radiosensitizers (RS), including: XRD-0394 (ATM/DNA-PK inhibitor, phase I, XRad Therapeutics, RS), lartesertib (ATM inhibitor, phase I, Merck, MT), M-3541 (ATM inhibitor, phase I, Merck, RS), AZD-1390 (ATM inhibitor, phase I, AstraZeneca, RS+CS), CC-115 (DNA-PK/mTOR inhibitor, phase II, Celgene, MT+RS+CS), BR-101801 (DNA-PK/PI3Kγ inhibitor, phase I, Boryung, MT), M-9831 (DNA-PK inhibitor, phase I, Merck and Vertex, MT+CS), panulisib (DNA-PK/mTOR/PI3Kα/ALK1 inhibitor, phase I, Piramal Enterprises and Piramal Life Sciences, MT), peposertib (DNA-PK inhibitor, phase I, Merck, RS+MT+CS). There are no molecules of this class on the pharmaceutical market. These compounds are being tested against oncological diseases, including: liposarcoma, brain cancer, glioblastoma, non-small cell lung cancer, head and neck cancer, skin cancer, prostate cancer, chronic lymphocytic leukemia, lymphoma (B- and T-cell), hematological blood cancer, neuroendocrine cancer, adenocarcinoma, bladder cancer, breast cancer and colorectal cancer. One of the features of some molecules in this class is the ability to penetrate the blood-brain barrier (BBB), for example, AZD-1390, which may lead to the development of CNS-targeted chemo- and radiosensitizers.

Chinese Published Application No. CN 103936762 A, IPC C07D 498/04, A61K 31/5383, A61P 35/00, A61P 35/02, July 23, 2014, discusses the synthesis and biological testing results for N- (5-(3,4-dihydro-2 H- [1,4]oxazino[3,2-c]quinolin-9-yl)pyridin-3-yl)sulfonamide derivatives as inhibitors of mTOR and PI3K kinases, including their 3-oxo derivatives. Various esters are also indicated as possible substituents at position 2 of the pyridine ring. However, activity against ATM, DNA-PK, and ATR kinases is not provided. Moreover, the examples present only 2-methoxypyridyl and 2-chloropyridyl derivatives. Considering that within the framework of the presented application we obtained selective inhibitors of ATM (IC ₅₀ <10 nM) and DNA-PK (IC ₅₀ <10 nM), which did not show activity against mTOR (IC ₅₀ >1 μM, IC _max =2,000) and PI3K isoforms (IC ₅₀ >1 μM, IC _max >10,000), it is not obvious that the structures described in application CN 103936762 A will inhibit the enzymatic activity of ATM, DNA-PK and ATR and vice versa. In our case, it was shown that 4- N -Me (R ³ =Me) and 5- N (Z ₁ =N - cinnoline) derivatives affect the selectivity profile of molecules of this class. Furthermore, cinnoline derivatives are not declared in CN 103936762 A. The influence of the methyl group on selectivity has been previously noted in a series of 1 H -imidazo[4,5-c]quinolin-2(3 H )-ones (Pike KG et al. J Med Chem 2018; 61: 3823). For the example molecule (8-(6-(3-(dimethylamino)propoxy)pyridin-3-yl)-3-methyl-1-(tetrahydro-2 H -pyran-4-yl)-1 H -imidazo[4,5-c]quinolin-2(3 H )-one), the following activities (IC ₅₀ μM) were obtained: 0.00004 (ATM), 0.14 (DNA-PK), 0.20 (mTOR), 0.32 (PI3Kα), 1.8 (PI3Kβ), 1.1 (PI3Kγ), and 0.27 (PI3Kδ). Despite the high activity against ATM kinase, the molecule inhibited the activities of mTOR, PI3Kα, and PI3Kδ relatively strongly.

Published international application No. WO 2020052688 A1, IPC C07D 487/04, A61K 31/5025, A61P 35/00, March 19, 2020, presents derivatives of 8-(pyridin-3-yl)-1 H -imidazo[4,5-c]cinnolin-2(3 H )-ones as selective ATM kinase inhibitors. In particular, cinnoline analogs substituted with fluorine at position 7 in the 8-(pyridin-3-yl)-1 H -imidazo[4,5-c]cinnolin-2(3 H )-one substructure were shown to selectively inhibit ATM kinase activity (IC ₅₀ = 1.9 nM), while the activity towards ATR, PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ and mTOR was greater than 10 μM. However, WO 2020052688 A1 discusses exclusively fluorine-substituted at position 7 1 H -imidazo[4,5-c]quinolin-2(3 H )-ones or 1 H -imidazo[4,5-c]cinnolin-2(3 H )-ones. The authors note that the cinnoline modification reduces the affinity of molecules for aldehyde oxidase, which prevents their metabolism, but do not associate it with selectivity.

Published international application No. WO 2021022078 A1, IPC A61K 31/444, A61K 31/4545, A61K 31/4745, C07D 471/04, C07D 471/10, C07D 491/20, 04.02.2021, presents ATM and DNA-PK inhibitors of the class of spiro -derivatives of N- (5-(2-oxo-2,3-dihydro-1 H -pyrrolo[2,3-c]quinolin-8-yl)pyridin-3-yl)sulfonamide, where the introduction of a fluorine atom at position 7 of 1 H -pyrrolo[2,3-c]quinolin-2(3 H )-one resulted in a significant loss of activity against mTOR. However, for the molecules given in the examples, the inhibitory activity towards PI3Kα/δ was retained (mainly IC ₅₀ values <100 nM), indicating a relatively low selectivity of this class of molecules. Examples of molecules containing cinnoline in their structure have not been described. We have shown that comparable selectivity can be obtained without the fluorine atom in the analogous position in the structure of 9-(pyridin-3-yl)-2 H -[1,4]oxazino[3,2-c]cinnolin-3(4 H )-ones.

Published international application No. WO 2019201283 A1, IPC C07D 471/10, A61K 31/444, A61K 31/4545, A61K 31/4745, A61K 31/502, A61P 35/00, 24.10.2019, describes spiro -derivatives of 1 H -pyrrolo[2,3-c]quinolin-2(3 H )-ones, 1,2-dihydrobenzo[ f ][1,7]naphthyridine-3(4 H )-ones and 1,2-dihydropyrazino[2,3-c]quinolin-3(4 H )-ones, as well as their cinnoline analogs, as inhibitors of ATM and DNA-PK kinases. Although many spiro derivatives of 1 H -pyrrolo[2,3-c]quinolin-2(3 H )-ones showed activity against ATM and DNA-PK kinases (IC ₅₀ < 0.5 nM), their six-membered analogs: 1 H -spiro[benzo[ f ][1,7]naphthyridine-2,1'-cyclobutan]-3(4 H )-ones (one example in the application) and 1' H -spiro[cyclopropan-1,2'-pyrazino[2,3-c]quinolin]-3'(4' H )-ones (two examples in the application) did not show activity against ATM and DNA-PK kinases (IC ₅₀ ≥ 100 nM). Furthermore, WO 2019201283 A1 does not claim 2 H -[1,4]oxazino[3,2-c]quinolin-3(4 H )-ones and 2 H -[1,4]thiazino[3,2-c]quinolin-3(4 H )-ones, nor their cinnoline analogues, and selectivity for ATR, PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ and mTOR for example molecules in WO 2019201283 A1 is not presented. In Chinese Published Application No. CN 109705139 A, IPC C07D 498/04, 03.05.2019, halogenated 9-position 2 H- [1,4]oxazino[3,2-c]quinolin-3(4 H )-one derivatives are described as inhibitors of c-Met kinase activity, without mentioning ATM and/or DNA-PK kinases.

Published international application No. WO2009155527, IPC C07D 487/04, C07D 487/02, C07D 403/02, A61K 31/437, A61P 35/00, 23.12.2009, presents substituted quinolines as multikinase inhibitors. In particular, 1,2-dihydrobenzo[ f ][1,7]naphthyridin-3(4 H )-ones and 1,2-dihydropyrazino[2,3-c]quinolin-3(4 H )-ones fall under the Markush structure. However, activity against ATM, ATR and DNA-PK is not indicated in the application. Furthermore, 2 H -[1,4]oxazino[3,2-c]quinolin-3(4 H )-ones and 2 H -[1,4]thiazino[3,2-c]quinolin-3(4 H )-ones do not fall within the Markush structure provided in the application. Taking into account the above, it can be stated that the structures claimed in the present invention are new, their activity against the target kinases ATM and DNA-PK, as well as the off-target kinases ATR, PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ and mTOR is not obvious and does not clearly follow from previously published patent applications and scientific articles.

The aim of the present invention is to develop and create new heterocyclic selective dual inhibitors of ATM and DNA-PK kinases that are promising in clinical practice for the treatment and/or prevention of oncological diseases as monotherapy and/or in combination with radiotherapy, chemotherapy and/or immunotherapy.

The technical result of the invention is the activity of chemical compounds in relation to ATM and DNA-PK kinases, and an increase in selectivity in relation to related kinases.

The specified technical result is achieved through the development and creation of compounds of the general formula (I):

or a corresponding pharmaceutical composition or salt, wherein:

- p and q are chosen independently and are equal to 0, 1, 2 or 3;

- B is selected independently and is a fused aromatic or non-aromatic carbocycle or heterocycle;

- M ₁ and M ₂ are selected independently and represent hydrogen or M ₁ and M ₂ together form =O (keto group);

- X, W and V are selected independently and are hydrogen, halogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted haloalkyl, optionally substituted alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl, -R ₆ -CN, -R ₆ -NO ₂ , -R ₆ -OR ₅ , -R ₆ -N(R ₄ )R ₅ , -OR ₆ -N(R ₄ )R ₅ , -N=C(R ₄ )R ₅ , -S(O) _r R ₄ , -OS(O) ₂ CF ₃ , -R ₆ -C(O)R ₄ , -C(S)R ₄ , -R ₆ -C(O)OR ₄ , -C(S)OR ₄ , -R ₆ -C(O)N(R ₄ )R ₅ , -C(S)N(R ₄ )R ₅ , -N(R ₅ )C(O)R ₄ , -N(R ₅ )C(S)R ₄ , -N(R ₅ )C(O)OR ₄ , -N(R ₅ )C(S)OR ₄ , -N(R ₅ )C(O)N(R ₄ )R ₅ , -N(R ₅ )C(S)N(R ₄ )R ₅ , -N(R ₅ )S(O) _t R ₄ , -N(R ₅ )S(O) _t N(R ₄ )R ₅ , -R ₆ -S(O) _t N(R ₄ )R ₅ , -OP(O)(R ₄ )R ₅ , -OP(O)R ₄ O(R ₄ ), -OP(O)R ₄ N(R ₄ )R ₅ , -N(R ₅ )-P(O)(R ₄ )R ₅ , -N(R ₅ )-P(O)R ₄ O(R ₄ ), -N(R ₅ )-P(O)R ₄ N(R ₄ )R ₅ , -N(R ₅ )-P(O)O(R ₄ )N(R ₄ )R ₅ , -N(R ₅ )-P(O)N(R ₄ )R ₅ N(R ₄ )R ₅ , -N(R ₅ )C(=NR ₅ )R ₄ , -N(R ₅ )C(=NR ₅ )N(R ₄ )R ₅ , and -N(R ₅ )C(=N-CN)N(R ₄ )R ₅ , where r is independently selected and equal to 0, 1, or 2, t is independently selected and equal to 1 or 2; or two adjacent substituents V, or W, or X, together with the carbon atoms of the ring to which they are directly bonded, form a fused aromatic or non-aromatic carbocycle or heterocycle;

- Y is independently selected and represents -O-, -S- or -SO ₂ -;

- Z ₁ and Z ₂ are selected independently and represent -C(R _1a )- or N;

- R _1a is independently selected and is hydrogen, alkyl, halogen, -CN, -NR ₂ R ₄ or OR ₂ ;

- R ₂ , R ₄ and R ₅ are selected independently and represent hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted haloalkyl, optionally substituted alkoxyalkyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heterocyclyl, optionally substituted heteroaryl;

- R ₃ is independently selected and is hydrogen or alkyl;

- R ₆ is selected independently and represents a direct bond or a linear or branched optionally substituted alkylene chain, a linear or branched optionally substituted alkenylene chain, a linear or branched optionally substituted alkynylene chain, or an optionally substituted heterocyclylene.

According to the present invention, pharmaceutically acceptable carriers, vehicles, or excipients include, but are not limited to, any adjuvant, vehicle, filler, lubricant, sweetener, diluent, preservative, colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier approved for use in humans or domestic animals.

Pharmaceutically acceptable salts include, but are not limited to, acid addition salts or base addition salts.

Pharmaceutically acceptable acid addition salts include salts that retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable and which are formed with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., and organic acids such as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinous acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, Pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid, undecylenic acid, etc.

Pharmaceutically acceptable base addition salts include salts that retain the biological effectiveness and properties of the free acids, but are not biologically or otherwise undesirable. These salts are prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol, N,N -dimethylaminoethanol, N,N -diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benetamine, benzathine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins and the like. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

Crystallization often results in the formation of a solvate of the compound of the invention. As used herein, the term "solvate" refers to an aggregate that contains one or more molecules of the compound of the invention with one or more molecules of a solvent. The solvent may be water, in which case the solvate may be a hydrate. In some cases, the solvent may be an organic solvent. Thus, the compounds of the present invention may exist as hydrates, including the monohydrate, dihydrate, hemihydrate, sesquihydrate, trihydrate, tetrahydrate, and the like, as well as the corresponding solvated forms. The compound of the invention may be true solvates, while in other cases, the compound of the invention may simply retain extraneous water or be a mixture of water and some extraneous solvent.

A pharmaceutical composition includes a preparation of the compound that is the subject of the invention and a vehicle generally accepted in the art for delivering the biologically active compound to mammals, such as humans. Such a vehicle includes all pharmaceutically acceptable carriers, diluents, or excipients.

Among the compounds of formula ( I ), the following compounds are preferred:

Connection number ID Structure Molecular weight 1 RA-O-00017 547.63 2 RA-O-00018 511.6 3 YAI-22-0025 548.62 4 YAI-22-0025F 566.61 5 YAI-22-0025a 530.58 6 RA-O-00020 533.6 7 RA-O-00019 533.65 8 RA-S-00016 527.66 9 YAI-22-0023 559.65 10 YAI-22-0024 534.59

Compounds of formula ( I ) are inhibitors of the ATM and DNA-PK kinase family.

The invention includes the use of a compound of the present invention for inhibiting the activity of ATM and DNA-PK kinases in cells or in a subject, or in the treatment or prevention of conditions associated with the activity or expression of ATM and DNA-PK kinases.

The compounds of the present invention can be used to treat or prevent cancer in a mammal, preferably a human, wherein the method of treating cancer according to the present invention comprises administering a therapeutically effective amount of the compound of the invention to the body of a mammal in need of stopping, slowing, or reversing the growth, development, or spread of cancer, including solid tumors or other forms of cancer. In some embodiments, the compound is administered to a mammal receiving radiotherapy.

In another aspect, the invention provides methods for treating or preventing cancer in mammals, the methods comprising administering to a mammal in need thereof a therapeutically effective amount of a compound of the invention. In some embodiments, the compound is administered to the mammal in combination with a DNA-damaging agent. Examples of DNA-damaging agents include cisplatin, oxaliplatin, carboplatin, valrubicin, idarubicin, calicheamicin, PARP inhibitors, etc.

In another aspect, the invention provides pharmaceutical compositions comprising the compounds of the invention and pharmaceutically acceptable excipients. In one embodiment, the pharmaceutical composition comprises the compound of the invention in a pharmaceutically acceptable carrier and in an amount effective for the treatment of an oncological disease in a mammal.

The compound of the invention, when used in combination therapy, may enhance the effectiveness of another drug treatment or may reduce the incidence and/or severity of adverse events associated with the other drug treatment. For example, side effects of radiotherapy (e.g., oral or gastrointestinal mucositis, dermatitis, pneumonitis, or fatigue) may be reduced in patients receiving combination therapy comprising the compound of the invention and radiotherapy (e.g., the incidence of side effects may be reduced by at least 1%, 5%, 10%, or 20%) relative to patients receiving radiotherapy without the compound of the invention. In addition, other adverse events that may be reduced in patients receiving combination therapy comprising a compound of the invention and radiotherapy (e.g., the incidence of adverse events may be reduced by at least 1%, 5%, 10%, or 20%) relative to patients receiving radiotherapy without the compound of the invention may include late effects of radiotherapy, such as radiation-induced pulmonary fibrosis, cardiac damage, bowel obstruction, nerve damage, vascular damage, lymphedema, brain necrosis, or radiotherapy-induced cancer. Similarly, when the compound is administered in combination therapy with another anticancer drug (e.g., as described herein), the combination therapy may cause the same or even increased tumor cell death, even if the dose of the other anticancer drug is reduced. Thus, reducing the dosage of other anticancer drugs may reduce the severity of adverse events caused by other anticancer drugs.

In another aspect, the present invention is directed to the use of the compounds of the invention, as set forth above, in the form of their stereoisomer, enantiomer, tautomer, or mixtures thereof, or a pharmaceutically acceptable salt or solvate thereof, or the use of a pharmaceutical composition comprising a pharmaceutically acceptable excipient and a compound of the invention, as set forth above, in the form of its stereoisomer, enantiomer, tautomer, or mixtures thereof, or a pharmaceutically acceptable salt or solvate thereof, in the preparation of a medicament for use in the treatment of a disease. In some embodiments, the compound of the invention is administered in combination with radiotherapy. In other embodiments, the compound of the invention is administered in combination with a DNA damaging agent. In some embodiments, the disease is cancer.

Examples of cancer types treatable using the methods and uses disclosed herein include, but are not limited to, leukemias and lymphomas - acute myeloid leukemia, acute lymphoblastic leukemia, acute megakaryocytic leukemia, promyelocytic leukemia, erythroleukemia, lymphoblastic T-cell leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, hairy cell leukemia, chronic neutrophilic leukemia, plasmacytoma, immunoblastic large cell leukemia, mantle cell leukemia, multiple myeloma, malignant lymphoma, diffuse large B-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lymphoblastic T-cell lymphoma, Burkitt's lymphoma, and follicular lymphoma.

Examples of cancer types that can be treated using the methods and uses disclosed herein include, but are not limited to, brain cancer (e.g., astrocytoma, glioma, glioblastoma, medulloblastoma, ependymoma), bladder cancer, breast cancer, central nervous system tumors, cervical cancer, rectal cancer, colon cancer, endometrial cancer, esophageal cancer, gastrointestinal stromal tumor, gastric cancer, head and neck cancer, oral cancer, hepatocellular carcinoma, cholangiocarcinoma, metastatic liver disease, Merkel cell carcinoma, lung cancer, melanoma, mesothelioma, nasopharyngeal cancer, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, salivary gland cancer, sarcomas, testicular cancer, urothelial cancer, vulvar cancer, and Wilms' tumor.

In additional embodiments, examples of cancers treatable using the methods and applications disclosed herein are not limited to metastases and metastatic cancer. For example, the methods and applications disclosed herein for treating cancer may include the treatment of both primary tumors and metastases.

In some embodiments, the methods and uses disclosed herein involve pre-administering an ATM and DNA-PK inhibitor to a subject prior to radiation therapy or a DNA-damaging agent. Pre-administration of a dual ATM and DNA-PK inhibitor can delay or eliminate DNA damage repair following radiation therapy.

Radiotherapy includes, but is not limited to, external beam radiotherapy using x-rays (photons), cobalt-60 gamma rays or other radioactive isotopes, neutrons, electrons, protons, carbon ions, helium ions, and other charged particles. Radiotherapy also includes brachytherapy and radiopharmaceuticals that emit gamma rays, alpha particles, beta particles, Auger electrons, or other types of radioactive particles from isotopes including iridium-192, iodine-125, cesium-137, palladium-103, phosphorus-32, yttrium-90, gallium-67, astatine-211, radium-223, and other radioactive isotopes. Radiotherapy also includes radioimmunotherapy (RIT) with antibodies or small molecules that are conjugated to radioactive isotopes, including iodine-131, yttrium-90, actinium-225, astatine-211, gallium-67, and other radioactive isotopes.

In some embodiments, the combination therapy comprises administering to the subject an ATM and DNA-PK inhibitor and an anti-tumor agent, such as cisplatin, oxaliplatin, carboplatin, topoisomerase I inhibitors, topoisomerase II inhibitors, anthracyclines, valrubicin, idarucin, calicheamicin, PARP inhibitors (e.g., olaparib, rucaparib, niraparib, veliparib, talazoparib), and other anti-tumor agents known to those skilled in the art.

In some embodiments, the combination therapy comprises administering to the subject an ATM and DNA-PK inhibitor and anti-tumor immunotherapeutic agents, including, but not limited to, ipilimumab, ofatumumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, etc.

In the combination therapy described herein, an ATM and DNA-PK inhibitor may be administered to a subject concurrently or sequentially (e.g., before or after) another drug or radiotherapy.

Organic synthesis

The compounds of this invention can be prepared using the synthetic methods described below. These methods are not exhaustive and allow for reasonable modifications. The reactions described should be carried out using suitable solvents and materials. When implementing these general procedures for the synthesis of specific substances, it is necessary to consider the functional groups present in the substances and their influence on the reaction. To obtain certain substances, it is necessary to change the order of the steps or to choose one of several alternative synthesis schemes. It should be understood that these and all examples provided in the application materials are not limiting and are provided solely to illustrate the present invention.

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3-oxo-3,4-dihydro- 2H -[1,4]oxazino[3,2-c]quinolin-9-yl)pyridin-3-yl)benzenesulfonamide (RA-O-00017):

Compound 3. To a solution of 2.17 g (0.021 mol, 1.25 equiv.) of alcohol ( 2 ) in 50 ml of "dry" THF at 0°C was added portionwise 0.95 g of 80% NaH (0.032 mol, 1.9 equiv.), the mixture was stirred for 15 min and a solution of 4 g (0.017 mol, 1 equiv.) of compound ( 1 ) in 25 ml of THF was added dropwise. The mixture was stirred for 12 h, the reaction mixture was poured into 200 ml of water and extracted with EA (3 times 100 ml). The combined organic phases were dried over anhydrous Na ₂ SO ₄ and evaporated. The resulting residue was purified by chromatography on silica gel, el-nt QM - QM-MeOH 5%. MS m/z 305.2 (M ⁺ H ⁺ ). Yield 3 was: 1 g, 16%.

Compound 4. Compound ( 3 ) 1 g (0.0033 mol, 1 equiv.) was dissolved in 50 ml of a 4:1 mixture of acetic acid and water, and 0.74 g (0.0132 mol, 4 equiv.) of iron powder was added portionwise. The reaction mixture was stirred for 12 hours. 50 ml of EA was added to the reaction mixture, filtered through a zeolite layer, evaporated, dissolved in EA, and filtered again through a zeolite layer. The filtrate was washed with a K ₂ CO ₃ solution, dried over anhydrous Na ₂ SO ₄ , and evaporated. The resulting residue was used in the next step without further purification. The yield of 4 was: 0.8 g, 88%.

Compound 6. To a solution of 0.3 g (0.0011 mol, 1 equiv.) of compound ( 4 ) in 25 ml of pyridine were added 10 mg of DMAP and 0.58 g (0.0033 mol, 3 equiv.) of sulfonyl chloride ( 5 ), and the mixture was stirred at 80°C for 24 hours. Pyridine was evaporated, the residue was poured into water and extracted with EA (3 times 100 ml). The combined organic phases were dried over _{anhydrous Na2SO4} and evaporated. The resulting residue was purified by chromatography on silica gel, el-nt XM-MeOH 25%. MS m/z 415.4 (M ⁺ H ⁺ ). The yield of 6 was: 0.4 g, 88%.

Compound 8. To a solution of 0.5 g (0.00185 mol) of compound ( 7 ) in 50 ml of MeOH, 0.5 g of Raney nickel was added, and 0.8 ml of hydrazine hydrate was added dropwise. The mixture was stirred for 12 hours, then 50 ml of MeOH was added to the reaction mixture, the mixture was stirred for 15 minutes, then filtered through a layer of zeolite, and the filtrate was evaporated. The residue was re-evaporated twice with toluene. 380 mg (yield 86%) of compound ( 8 ) was obtained, which was immediately used in the next step.

Compound 9. To a solution of 0.38 g (0.00158 mol, 1 equiv.) of compound ( 8 ) in 50 ml of DMF was added 0.83 g (0.00347, 2 equiv.) _{of K2CO3} , and then 0.24 g (0.0019 mol, 1.2 equiv.) of chloroacetyl chloride were added dropwise. The mixture was stirred at 80°C for 12 hours, then the reaction mixture was poured into 200 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 280.2 (M ⁺ H ⁺ ). ^1H NMR (400 MHz, DMSO _-d6 ) δ 11.12 (s, 1H), 8.52 (s, 1H), 8.09 (d, J = 1.7 Hz, 1H), 7.86 (d, J = 9.0 Hz, 1H), 7.74 (dd, J = 9.0, 1.8 Hz, 1H), 4.92 (s, 2H). Compound 9 was obtained (0.28 g, 63% yield).

Compound 10. To a solution of 0.2 g (0.00071 mol, 1 equiv.) of compound ( 9 ) in 25 ml of DMF was added 0.043 g (0.0142 mol, 2 equiv.) of NaH, then 0.126 g (0.00088 mol, 1.25 equiv.) of _CH3I were added dropwise. The mixture was stirred for 1 hour, the reaction mixture was poured into 100 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 294.2 (M ⁺ H ⁺ ). Compound 10 (0.16 g, yield 76%) was obtained.

CompoundRA-O-00017.To a solution of 0.15 g (0.51 mmol, 1 equiv.) of compound (10) 0.136 g (0.54 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.16 g (1.63 mmol, 3.2 equiv.) KOAc. The solution was degassed by passing a current of Ar for 10 minutes and 30 mg (0.041 mmol, 0.08 equiv.) Pd(dppf)Cl were added.₂. The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.35 g (1 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.185 g (0.44 mmol, 0.88 equiv.) of the compound (6). The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol, 0.08 equiv.) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 10% Methylpropional (MeOH). MS m/z 548.6 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 9.94 (s, 1H), 9.50 (s, 1H), 8.89 (s, 1H), 8.42 (s, 1H), 8.08 (s, 2H), 7.98-7.93 (m, 2H), 7.77-7.60 (m, 5H), 5.10 (s, 2H), 4.14 (s, 2H), 3.48 (s, 3H), 3.19 (s, 2H), 2.82 (s, 6H), 1.93 (s, 2H). Received connectionRA-O-00017(65 mg, yield 27%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3-oxo-3,4-dihydro-2 H -[1,4]oxazino[3,2-c]quinolin-9-yl)pyridin-3-yl)cyclopropanesulfonamide (RA-O-00018):

Compounds 4 and 10 were prepared according to the procedure described for RA-O-00017.

Compound 12. To a solution of 0.35 g (0.00127 mol, 1 equiv.) of compound ( 4 ) in 25 ml of pyridine were added 10 mg of DMAP and 0.53 g (0.0038 mol, 3 equiv.) _of sulfonyl chloride ( 11 ), then stirred at 80°C for 24 h. Pyridine was evaporated, the residue was treated with 5% _K2CO3 solution and extracted with EA (3 times 100 ml). The combined organic phases were dried over _{anhydrous Na2SO4} and evaporated. The resulting residue was purified by chromatography on silica gel, el-nt QM-MeOH 25%. MS m/z 379.4 (M ⁺ H ⁺ ). The yield of 12 was: 0.4 g (83%).

CompoundRA-O-00018.To a solution of 0.15 g (0.51 mmol, 1 equiv.) of compound (10) 0.136 g (0.54 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.16 g (1.63 mmol, 3.2 equiv.) KOAc. The solution was degassed by passing a current of Ar for 10 minutes and 30 mg (0.041 mmol, 0.08 equiv.) Pd(dppf)Cl were added.₂. The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.35 g (1 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.168 g (0.44 mmol, 0.88 equiv.) of the compound (12). The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol, 0.08 equiv.) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 10% Methylpropional (MeOH). MS m/z 512.4 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 9.54 (s, 1H), 9.35 (s, 1H), 8.89 (s, 1H), 8.48 (d, J = 1.7 Hz, 1H), 8.20 (s, 1H), 8.14 - 8.00 (m, 3H), 5.07 (s, 2H), 4.44 (t, J= 5.6 Hz, 2H), 3.47 (s, 3H), 3.35 (s, 2H), 2.78 (m, 7H), 2.16 (s, 2H), 1.06-0.87 (m, 4H). 55 mg of the compound were obtained.RA-O-00018(yield 24%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3,4-dihydro-2 H -[1,4]oxazino[3,2-c]quinolin-9-yl)pyridin-3-yl)benzenesulfonamide (RA-O-00019):

Compounds 8 and 6 were prepared according to the procedure described for RA-O-00017.

Compound 13. To a solution of 0.65 g (0.0027 mol, 1 equiv.) of compound ( 8 ) in 50 ml of DMF were added 2.44 g (0.017 mol, 6 equiv.) of K ₂ CO ₃ , then 1.5 g (0.008 mol, 3 equiv.) of dibromoethane were added dropwise. The mixture was stirred at 80 °C for 12 hours, then the reaction mixture was poured into 200 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 266.2 (M ⁺ H ⁺ ). The yield of 13 was: 0.63 g (87%).

Compound 14. To a solution of 0.35 g (0.0013 mol, 1 equiv.) of compound ( 13 ) in 30 ml of DMF was added 0.5 ml of 37% formaldehyde solution, then 2.4 g (0.011 mol, 9 equiv.) of sodium triacetoxyborohydride (STAB). The reaction mixture was stirred for 3 h, then washed with a saturated solution of NaHCO ₃ . The organic phase was dried over anhydrous Na ₂ SO ₄ and evaporated. The resulting residue was used in the next step without further purification. MS m/z 280.2 (M+H+). The yield of 14 was: 0.15 g (42%).

CompoundRA-O-00019.To a solution of 0.07 g (0.25 mmol, 1 equiv.) of compound (14) 0.068 g (0.27 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.08 g (0.8 mmol, 3.2 equiv.) KOAc. The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.02 mmol, 0.08 equiv.) Pd(dppf)Cl were added.₂. The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.17 g (1 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.093 g (0.44 mmol, 0.88 equiv.) of the compound (6). The solution was degassed by passing an Ar current for 10 minutes and 15 mg (0.02 mmol, 0.08 equiv.) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 5% Methylpropional-MeOH. MS m/z 534.6 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 8.59 (s, 1H), 8.18 (s, 1H), 7.90 - 7.85 (m, 2H), 7.81 - 7.73 (m, 3H), 7.67 - 7.52 (m, 5H), 4.57 (s, 2H), 4.17 (s, 2H), 3.32 (s, 2H), 3.15 (t, J= 7.0 Hz, 2H), 3.02 (s, 3H), 2.78 (s, 6H), 1.98 (s, 2H). 15 mg of the compound were obtained.RA-O-00019(yield 6%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(3-oxo-3,4-dihydro-2 H -[1,4]oxazino[3,2-c]quinolin-9-yl)pyridin-3-yl)benzenesulfonamide (RA-O-00020):

Compounds 10 and 6 were prepared according to the procedure described for RA-O-00017.

CompoundRA-O-00020.To a solution of 0.15 g (0.53 mmol, 1 equiv.) of compound (10) 0.136 g (0.53 mmol, 1 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.16 g (1.63 mmol, 3 eq.) of KOAc. The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol, 0.08 eq.) of Pd(dppf)Cl were added.₂. The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.35 g (1 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.185 g (0.44 mmol, 0.88 equiv.) of the compound (6). The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol, 0.08 equiv.) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 10% Methylpropional-MeOH. 70 mg of product was obtained, which was additionally recrystallized from EtOH, and the resulting precipitate was washed with diethyl ether. MS m/z 534.6 (M⁺H⁺).¹H NMR (400 MHz, DMSO-d ₆) δ 11.08 (s, 1H), 8.49 (s, 1H), 7.98 (s, 1H), 7.96 (d, J = 8.9 Hz, 1H), 7.87 (s, 1H), 7.84 - 7.72 (m, 3H), 7.63 - 7.48 (m, 4H), 5.00 (s, 2H), 4.25 (t, J= 5.6 Hz, 2H), 3.13 (t,J= 5.8 Hz, 2H), 2.80 (s, 6H), 2.05 (m, 2H). Compound outputRA-O-00020amounted to 20 mg (9%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3-oxo-3,4-dihydro-2 H -[1,4]thiazino[3,2-c]quinolin-9-yl)pyridin-3-yl)cyclopropanesulfonamide (RA-S-00016):

Compound 12 was prepared according to the procedure described for RA-O-00017.

Compound 17. To a solution of 0.225 g (0.00187 mol, 1.2 equiv.) of compound ( 16 ) in 30 ml of acetone was added 0.26 ml of _Et3N (0.00187 mol, 1.2 equiv.) and then 0.45 g (0.00156 mol, 1 equiv.) of compound ( 15 ) was added portionwise. The reaction mixture was stirred for 12 h, the formed precipitate was filtered off, the filtrate was evaporated, poured with water and extracted with EA (3 times ₅₀ ml). The combined organic phases were dried over anhydrous _Na2SO4 and evaporated. The resulting residue was purified by chromatography on silica gel, el-nt toluene. MS m/z 372.4 (M ⁺ H ⁺ ). The yield of 17 was 0.3 g (52%).

Compound18.Connection170.3 g (0.00081 mol, 1 equiv.) was dissolved in 20 ml of acetic acid and 0.226 g (0.00404 mol, 5 equiv.) of iron powder was added in portions and stirred for 1 hour, then 12 hours at 70 °C. The resulting precipitate was filtered off, the precipitate was poured with dioxane, boiled, then filtered. The filtrate was purified by chromatography on silica gel without preliminary evaporation, el-nt dioxane, MS m / z 297.2 (M⁺H⁺). Exit18 amounted to 0.2 g (84%).

Compound 19. To a solution of 0.2 g (0.00068 mol, 1 equiv.) of compound 18 in 25 ml of DMF was added 0.024 g (0.00102 mol, 1.5 equiv.) of 60% NaH, and then 0.120 g (0.00085 mol, 1.25 equiv.) of _CH3I were added dropwise. The reaction mixture was stirred for 1 h, then poured into 100 ml of water, and the formed precipitate was filtered off. MS m/z 309.2 (M ⁺ H ⁺ ). 19 (0.14 g, 67%) was obtained.

CompoundRA-S-00016.To a solution of 0.1 g (0.32 mmol, 1 equiv.) of the compound190.86 g (0.34 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.095 g (0.97 mmol, 3.2 eq.) KOAc. The solution was degassed by passing an Ar current for 10 min and 19 mg (0.025 mmol, 0.08 eq.) Pd(dppf)Cl were added.₂The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.2 g (0.64 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.108 g (0.28 mmol, 0.88 equiv.) of the compound (12). The solution was degassed by passing an Ar current for 10 minutes and 19 mg (0.025 mmol, 0.08 equiv.) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 10% Methylpropional (MeOH). MS m/z 528.4 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 9.49 (s, 1H), 9.37 (s, 1H), 8.91 (s, 1H), 8.49 (d, J = 2.3 Hz, 1H), 8.11 (m, 3H), 8.03 (dd, J = 8.7, 2.0 Hz, 1H), 4.45 (t, J = 5.9 Hz, 2H), 3.76 (s, 2H), 3.55 (s, 3H), 3.35 (m, 2H), 2.84 (m, 6H), 2.79 (m, 1H), 2.16 (t, J = 7.8 Hz, 2H), 0.97 (m, 4H). Received connection RA-S-00016(15 mg, 4%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-1,1-dioxido-3-oxo-3,4-dihydro-2 H -[1,4]thiazino[3,2-c]quinolin-9-yl)pyridin-3-yl)cyclopropanesulfonamide (YAI-22-0023):

Compound 19 was prepared according to the procedure described for RA-O-00018. Compound 12 was prepared according to the procedure presented for compound RA-O-00017.

Compound 20. To a solution of 0.766 g (0.00247 mol) of compound ( 19 ) in 25 ml of CH ₂ Cl ₂ at 0°C was added portionwise meta-chloroperbenzoic acid (mCPBA), the mixture was stirred for 4 h. The reaction mixture was washed with 5% K ₂ CO ₃ solution, the organic phase was dried over anhydrous Na ₂ SO ₄ and evaporated. The resulting residue was purified by chromatography on silica gel, CM-MeOH 10%. MS m/z 360.4 (M ⁺ H ⁺ ). The yield of 20 was 0.275 g (31%).

Compound 21. Compound 20. 0.27 g (0.00075 mol, 1 equiv) was dissolved in 20 ml of acetic acid and 0.167 g (0.003 mol, 5 equiv) of iron powder was added portionwise and the mixture was stirred for 1 hour, then for 12 hours at 70°C. 50 ml of EA was added to the reaction mixture, the precipitate was filtered off. The filtrate was evaporated, the residue was dissolved in 100 ml and washed with 5% K ₂ CO ₃ solution, the organic phase was dried over anhydrous Na ₂ SO ₄ and evaporated. The resulting residue was purified by chromatography on silica gel, CM-MeOH 10%. MS m/z 342.4 (M ⁺ H ⁺ ). The yield of 21 was 0.097 g (37%).

CompoundYAI-22-0023.To a solution of 0.09 g (0.26 mmol, 1 equiv.) of compound (21) 0.070 g (0.27 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.083 g (0.84 mmol, 3.2 equiv.) KOAc. The solution was degassed by passing a current of Ar for 10 min and 19 mg (0.025 mmol) Pd(dppf)Cl were added₂The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.17 g (0.52 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.10 g (0.26 mmol, 1 equiv.) of the compound (12). The solution was degassed by passing an Ar current for 10 minutes and 19 mg (0.025 mmol) of Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, eluent 10% CM-MeOH (MS m/z 560.4 (M⁺H⁺)), then on a preparative chromatograph in both acidic and neutral media. The compound was obtainedYAI-22-0023(12 mg, 14%). Spectrum¹H NMR of the obtained compound corresponded to¹H NMR spectrum for the RA-S-00016 molecule. Compound YAI-22-0023 has relatively low stability.

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3-oxo-3,4-dihydro-2 H -[1,4]oxazino[3,2-c]cinnolin-9-yl)pyridin-3-yl)benzenesulfonamide (YAI-22-0025):

Compound 23. A solution of 6.16 g (0.034 mol) of N -bromosuccinimide (NBS) in 10 ml of dry acetonitrile was added dropwise to a solution of 3.6 g (0.0266 mol) of compound ( 22 ) in 50 ml of dry acetonitrile at 0°C. The mixture was stirred at room temperature for 12 hours, the reaction mixture was evaporated, and the residue was purified by column chromatography on silica gel, eluent CH ₂ Cl ₂ . MS m/z 215.06 (M ⁺ H ⁺ ). The yield of 23 was 5.3 g (93%).

Connection24.To a solution of 5.3 g (0.0247 mol) of compound (23) a solution of 1.7 g (0.0247 mol) of NaNO was added dropwise to 30 ml of aqueous concentrated HCl at 0°C₂in 3 ml of water. The mixture was stirred at room temperature for 12 hours, then heated at reflux for 6 hours. The reaction mixture was cooled, the precipitate was filtered off, dried, and then washed with diethyl ether. MS m/z 226.06 (M⁺H⁺). Exit 24amounted to 4 g (72%).

Compound 25. 4 g (0.0177 mol) of compound ( 24 ) were added to 40 ml of propionic acid and stirred at 110°C for 30 min. Then 5 ml of concentrated _HNO3 were added dropwise, and the reaction mixture was stirred at 110°C for 2 h. Then heating was stopped and the mixture was stirred for 12 h. The formed precipitate was filtered off and dried over solid NaOH in a desiccator. MS m/z 271.2 (M ⁺ H ⁺ ). Compound 25 (2.7 g, 56%) was obtained.

Compound 26. To a solution of 2.7 g (0.001 mol) of compound ( 25 ) in 50 ml of MeOH, 0.5 g of Raney nickel was added, and 0.8 ml of hydrazine hydrate was added dropwise. The reaction mixture was stirred for 12 hours, then 50 ml of MeOH was added, and the mixture was stirred for 15 minutes, then filtered through a layer of zeolite, and the filtrate was evaporated. The residue was re-evaporated twice with toluene. Compound 26 (0.73 g, 30%) was obtained, which was used immediately in the next step.

Compound 27. To a solution of 0.73 g (0.003 mol, 1 equiv.) of compound ( 26 ) in 50 ml of DMF were added 1.57 g (0.0114, 3.8 equiv.) of K ₂ CO ₃ and then 0.4 g (0.0036 mol, 1.2 equiv.) of chloroacetyl chloride were added dropwise. The mixture was stirred at 80°C for 12 h, the reaction mixture was poured into 200 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 281.2 (M ⁺ H ⁺ ). Compound 27 (0.6 g, 70%) was obtained.

Compound 28. To a solution of 0.6 g (0.0021 mol, 1 equiv.) of compound ( 27 ) in 25 ml of DMF was added 0.15 g (0.0042 mol, 2 equiv.) of NaH, and then 0.377 g (0.00267 mol, 1.25 equiv.) of CH ₃ I were added dropwise. The reaction mixture was stirred for 1 h, then poured into 100 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 295.2 (M ⁺ H ⁺ ). Compound 28 (0.14 g, 22%) was obtained.

CompoundYAI-22-0025.To a solution of 0.14 g (0.00047 mmol, 1 equiv.) of compound (28) 0.126 g (0.0005 mmol, 1.05 equiv.) of B was added to 15 ml of “dry” dioxane₂Pin₂and 0.15 g (0.0015 mmol, 3.2 equiv.) KOAc. The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol) Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.35 g (1 mmol) Cs was added.₂CO₃in 1.5 ml of water. Then 0.23 g (0.0057 mmol, 1.2 equiv.) of the compound (6). The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol) of Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e-nt CM-MeOH 10% and preparative chromatography in the presence of trifluoroacetic acid. MS m/z 549.4 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 9.96 (s, 1H), 9.45 (s, 1H), 8.49 (d, J= 2.3 Hz, 1H), 8.37 (d,J = 8.9 Hz, 1H), 8.12 - 8.02 (m, 2H), 7.99 (d, J = 2.3 Hz, 1H), 7.82 - 7.73 (m, 2H), 7.69 (t, J= 7.4 Hz, 1H), 7.60 (t,J = 7.6 Hz, 2H), 5.17 (s, 2H), 4.14 (t, J= 5.9 Hz, 2H), 3.62 (s, 3H), 3.28 - 3.10 (m, 2H), 2.82 (s, 6H), 2.01 - 1.81 (m, 2H). 12 mg of the compound were obtained.YAI-22-0025(CF₃COO^-). The yield was 7%.

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(3-oxo-3,4-dihydro-2 H -[1,4]oxazino[3,2-c][1,5]naphthyridin-9-yl)pyridin-3-yl)benzenesulfonamide (YAI-22-0024):

Compound 30. 7 g (0.0485 mol) of Meldrum's acid ( 29 ) in 25 ml of trimethyl orthoformate was heated at reflux for 2 h. The reaction mixture was evaporated, and the residue was re-evaporated twice with toluene. 9.22 g (yield 99%) of compound ( 30 ) was obtained, which was immediately used in the next step.

Compound 31. To a solution of 9 g (0.048 mol, 1 equiv.) of compound ( 30 ) in 50 ml of isopropanol was added 6 g (0.048, 1 equiv.) of 6-methoxypyridin-3-amine ( 30a ). The mixture was stirred at reflux for 2 h, the reaction mixture was cooled, and the formed precipitate was filtered off. MS m/z 293.2 (M ⁺ H ⁺ ). Compound 31 (9.4 g, 67%) was obtained.

Compound 32. 30 ml of diphenyl ether were heated to 220°C and 9 g (0.03 mol) of compound ( 31 ) were added. The mixture was stirred at this temperature for 15 min, then cooled and 30 ml of hexane was added. The formed precipitate was filtered off, washed with diethyl ether and dried. MS m/z 177.2 (M ⁺ H ⁺ ). Compound 32 (4.5 g, 83%) was obtained.

Compound 33. 4.5 g (0.0255 mol) of compound ( 32 ) were added to 50 ml of propionic acid and the mixture was stirred at 110°C for 30 min, then 5 ml of concentrated HNO ₃ were added dropwise. The reaction mixture was stirred at this temperature for 2 h, heating was stopped, and the mixture was stirred for 12 h. The formed precipitate was filtered off and dried over solid NaOH in a desiccator. MS m/z 222.2 (M ⁺ H ⁺ ). Compound 33 (1.1 g, 19.5%) was obtained.

Compound 34. 0.1 g of 10% Pd/C was added to a solution of 0.7 g (0.0031 mol) of compound ( 33 ) in 25 ml of methanol and the mixture was hydrogenated at atmospheric pressure for 3 hours. The reaction mixture was filtered through a layer of zeolite, the residue was washed with methanol, the filtrate was evaporated, and 0.56 g of compound ( 34 ) was obtained, which was immediately used in the next step.

Compound 35. To a solution of 0.56 g (0.0029 mol, 1 equiv.) of compound ( 34 ) in 50 ml of DMF was added 1.61 g (0.012, 4 equiv.) of K ₂ CO ₃ , then 0.4 g (0.0035 mol, 1.2 equiv.) of chloroacetyl chloride were added dropwise. The mixture was stirred at 80°C for 12 h, then poured into 200 ml of water, the formed precipitate was filtered off and recrystallized from ethanol. MS m/z 232.2 (M ⁺ H ⁺ ). Compound 35 (0.26 g, 38%) was obtained.

Compound 36. 2 ml of concentrated HBr were added dropwise to a solution of 0.26 g (0.00112 mol) of compound ( 35 ) in 6 ml of acetic acid, the reaction mixture was stirred at 50°C for 24 h, evaporated, the residue was poured into 25 ml of 10% _Na2CO3 solution, the formed precipitate _was filtered off and dried over NaOH in a desiccator. MS m/z 218.2 (M ⁺ H ⁺ ). Compound 36 (0.162 g, 66%) was obtained.

Compound 37. 0.16 g (0.00073 mol) of compound ( 36 ) was heated at reflux in 20 ml _{of POCl3} for 4 hours, then the reaction mixture was evaporated, the residue was poured into 25 ml of 10% _Na2CO3 solution, the formed precipitate _was filtered off and dried over NaOH in a desiccator. MS m/z 236.4 (M ⁺ H ⁺ ). Compound 37 (0.093 g, 53%) was obtained.

CompoundYAI-22-0024.To a solution of 0.136 g (0.000328 mmol, 1 equiv.) of compound (37) in 15 ml of “dry” dioxane was added 0.087 g (0.000344 mol, 1.05 equiv.) B₂Pin₂and 0.1 g (0.001 mol, 3.2 equiv.) KOAc. The solution was degassed by passing an Ar current for 10 min and 30 mg (0.041 mmol) Pd(dppf)Cl were added.₂. The mixture was then stirred at 100°C under argon for 12 hours, then cooled to room temperature and a solution of 0.35 g (1 mmol, 2 equiv.) Cs was added.₂CO₃in 1.5 ml of water. Then 0.067 g (0.00028 mmol, 0.88 equiv.) of the compound (6). The solution was degassed by passing an Ar current for 10 minutes and 30 mg (0.041 mmol) of Pd(dppf)Cl was added.₂The mixture was then stirred at 100°C under argon for 8 hours. The reaction mixture was diluted with 15 ml of EtOAc and filtered through a bed of zeolite. 30 ml of water was added to the filtrate. The organic layer was separated, and the aqueous layer was extracted with EtOAc twice. 30 ml. The organic phases were combined, washed with a saturated NaCl solution, dried over anhydrous Na₂SO₄and evaporated. The resulting residue was purified by chromatography on silica gel, e.g., 10% Methylpropional (MeOH). MS m/z 548.6 (M⁺H⁺),¹H NMR (400 MHz, DMSO-d ₆) δ 11.20 (s, 1H), 8.68 - 8.51 (m, 2H), 8.42 - 8.27 (m, 2H), 8.15 (d,J= 9.0 Hz, 1H), 7.83 (d,J = 7.7 Hz, 2H), 7.61 - 7.48 (m, 3H), 5.03 (s, 2H), 4.21 (t, J= 5.8 Hz, 2H), 3.09 (t,J= 6.9 Hz, 3H), 2.74 (s, 6H), 2.04 - 1.87 (m, 2H). 15 mg of the compound were obtained.YAI-22-0024. The yield was 10%.

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(4-methyl-3-oxo-3,4-dihydro-2 H -[1,4]oxazino[3,2-c]cinnolin-9-yl)pyridin-3-yl)cyclopropanesulfonamide (YAI-22-0025a):

Compound 28 was prepared according to the procedure described for YAI-22-0025, compound 12 was prepared according to the procedure described for RA-O-00017. Compound YAI-22-0025a was obtained by a cross-coupling reaction similar to YAI-22-0025. The following analytical characteristics were obtained for compound YAI-22-0025a: MS m/z 513.16 (M ⁺ H ⁺ ), ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.81 (s, 1H), 9.33 (br s, 1H), 8.3 (s, 1H), 8.35 (d, J = 7.2 Hz, 1H), 8.22 (d, J = 7.2 Hz, 1H), 8.17 (s, 1H), 7.75 (s, 1H), 5.21 (s, 2H), 4.22 (t, J = 5.8 Hz), 4.17-3.61 (m, 4H), 3.66 (s, 3H), 3.23 (br s, 2H), 2.33 (s, 6H), 1.92 (br s, 2H). 12 mg of compound YAI-22-0025a (CF ₃ COO ^- ) were obtained. The yield of compound YAI-22-0025F was 10 mg (10%).

Method for obtaining N -(2-(3-(dimethylamino)propoxy)-5-(8-fluoro-4-methyl-3-oxo-3,4-dihydro- 2H -[1,4]oxazino[3,2-c]cinnolin-9-yl)pyridin-3-yl)benzenesulfonamide (YAI-22-0025F):

Compound 39. To a solution of BCl ₃ (5.9 mL, 1 M) in DCM (5.9 mmol) was added dropwise 1-bromo-2-fluoro-4-nitrobenzene ( 38 ) (1 g, 5.26 mmol) in CHCl ₂ CHCl ₂ (10 mL) with stirring at 0 °C. Then MeCN (1 mL) and AlCl ₃ (0.8 g, 5.9 mmol) were added portionwise to the mixture. The resulting mixture was stirred at 120 °C for 16 h. After cooling the mixture, 2N aqueous HCl solution (13 mL) was added dropwise at 0 °C. The resulting mixture was stirred at 100 °C for 2 h, then poured into ice water and extracted with DCM (15 mL x 2). The combined organic phases were washed with brine, dried over _{anhydrous Na2SO4} , filtered, and concentrated. The residue was purified by column chromatography on silica gel. Compound 39 (0.27 g, 22%), MS m/z 232 (M ⁺ H ⁺ ) was obtained.

Compound 40 was prepared by analogy with the procedure described for compound 28 , using 39 as the starting reagent. Compound YAI-22-0025F was prepared under Suzuki-Miyaura reaction conditions by analogy with compound YAI-22-0025. The following analytical characteristics were obtained for compound YAI-22-0025F: MS m/z 567.2 (M ⁺ H ⁺ ), ¹ H NMR (400 MHz, DMSO- d ₆ ) δ 9.89 (s, 1H), 9.47 (br s, 1H), 8.5 (s, 1H), 8.48 (s, 1H), 8.10 (s, 1H), 7.82 (s, 1H), 7.82-7.5 (m, 5H), 5.21 (s, 2H), 4.22 (t, J = 5.8 Hz), 3.66 (s, 3H), 3.23 (br s, 2H), 2.33 (s, 6H), 1.92 (br s, 2H). 10 mg (10%) of compound YAI-22-0025F were obtained.

The compounds of the present invention can be prepared by the above methods or similar methods using appropriate starting materials according to the selected substituents and their positions.

Biological tests

The use of the compounds of the present invention as an inhibitor of ATM and DNA-PK kinases can be determined by the methods of the following examples, and the activity levels of the compounds can be determined by IC ₅₀ values (the concentration of the compound that results in 50% inhibition of the enzyme activity).

The pharmacological tests and results are summarized as follows:

(1) ATM kinase inhibitory activity assay

The inhibitory activity of the synthesized compounds against ATM was determined by fluorescence resonance energy transfer (FRET) and compared with the positive control, KU-55933.

ATM(h) was incubated in a buffer containing 30 nM GST-cMyc-p53 and Mg/ATP (concentration as needed). The reaction was initiated by adding the Mg/ATP mixture. After incubation for 30 min at room temperature, the reaction was stopped by adding a stop solution containing EDTA. Then, a detection buffer containing a d2-labeled monoclonal antibody against GST and a europium-labeled anti-phospho-Ser15 antibody against phosphorylated p53 was added. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence signal (HTRF) was determined using the formula HTRF = 10,000 x (Em665 nm/Em620 nm). IC ₅₀ values were calculated using the Hill equation and a standard dose-response curve.

(2) DNA-PK kinase inhibitory activity assay

The inhibitory activity of the synthesized compounds against DNA-PK was determined by fluorescence resonance energy transfer (FRET) and compared with the positive control, PI-103.

DNA-PK(h) was incubated in a buffer containing 50 nM GST-cMyc-p53 and Mg/ATP (concentration as needed). The reaction was initiated by adding the Mg/ATP mixture. After incubation for 30 min at room temperature, the reaction was stopped by adding a stop solution containing EDTA. Then, a detection buffer containing a d2-labeled monoclonal antibody against GST and a europium-labeled anti-phospho-Ser15 antibody against phosphorylated p53 was added. The plate was read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined using the formula HTRF = 10,000 x (Em665 nm/Em620 nm). IC ₅₀ values were calculated using the Hill equation and a standard dose-response curve.

Illustrative examples of compounds with high inhibitory activity against ATM and DNA-PK kinases are shown in Table 1.

Activity ranges

A IC ₅₀ < 10 nM

B 10 nM < IC ₅₀ < 100 nM

C 100 nM < IC ₅₀ < 1,000 nM

Table 1.

Connection number ID ATM DNA-PK 1 RA-O-00017 B B 2 RA-O-00018 B B 3 YAI-22-0025 A C 4 YAI-22-0025F A B 5 YAI-22-0025a A B 6 RA-O-00020 A A 7 RA-O-00019 C C 8 RA-S-00016 B B 9 YAI-22-0023 B B 10 YAI-22-0024 A C

(3) Analysis of the selectivity of compounds against related kinases of the PIKK and PI3K families

The compounds of the invention were tested for inhibitory activity (IC ₅₀ ) against related kinases of the PIKK and PI3K families.

ATR kinase inhibitory activity assay

The inhibitory activity of the synthesized compounds against ATR was determined by fluorescence resonance energy transfer (FRET) and compared with the positive control, VE-822.

ATR/ATRIP(h) was incubated in a buffer containing 50 nM GST-cMyc-p53 and Mg/ATP (concentration as needed). The reaction was initiated by adding the Mg/ATP mixture. After incubation for 30 minutes at room temperature, the reaction was stopped by adding a stop solution containing EDTA. Next, detection buffer containing a d2-labeled monoclonal antibody against GST and a europium-labeled anti-phospho-Ser15 antibody against phosphorylated p53 was added. The plate was then read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence signal (HTRF) was determined using the formula HTRF = 10,000 x (Em665 nm/Em620 nm). IC ₅₀ values were calculated using the Hill equation and a standard dose-response curve.

mTOR kinase inhibitory activity assay

The inhibitory activity of the synthesized compounds against mTOR was determined radiometrically and compared with the positive control, PI-103.

mTOR/FKBP12 (h) was incubated with 50 mM 2-(4-(2-hydroxyethyl)-1-piperazinyl)ethanesulfonic acid (HEPES), pH 7.5, 1 mM EDTA, 0.01% Tween 20, 2 mg/ml substrate, 10 μM FKBP12, 3 mM _MnCl2 , and [gamma- ^33P ]-ATP (specific activity and concentration as needed). The reaction was initiated by adding the Mn/ATP mixture. After incubation for 40 min at room temperature, the reaction was stopped by adding phosphoric acid to a concentration of 0.5%. An aliquot of the reaction mixture was then applied to a filter and washed four times for 4 min in 0.425% phosphoric acid and once in methanol before drying and scintillation counting. IC ₅₀ values were calculated using the Hill equation and a standard dose-response curve.

PI3K kinase inhibitory activity assay

The inhibitory activity of the synthesized compounds against PI3K kinases was determined by fluorescence resonance energy transfer (FRET) and compared with the positive control, PI-103.

PI3K (p110α/p85α)(h), PI3K (p110α(E542K)/p85α)(h), PI3K (p110β/p85α)(h), PI3K (p110δ/p85α)(h), and PI3K (p120γ)(h) were incubated in a buffer containing 10 μM phosphatidylinositol 4,5-bisphosphate and Mg/ATP (concentration as needed). The reaction was initiated by adding ATP solution or a Mg/ATP mixture. After incubation for 30 min at room temperature, the reaction was stopped by adding a stop solution containing EDTA and biotinylated phosphatidylinositol 3,4,5-triphosphate. Detection buffer containing europium-labeled anti-GST monoclonal antibodies, GST-labeled GRP1 PH domain, and streptavidin-allophycocyanin was then added. The plate was then read in time-resolved fluorescence mode, and the homogeneous time-resolved fluorescence (HTRF) signal was determined using the formula HTRF = 10,000 x (Em665 nm/Em620 nm). _IC50 values were calculated using the Hill equation and a standard dose-response curve.

The activities of illustrative example molecules towards ATR, mTOR and PI3K kinase isoforms are given in Table 2.

Activity ranges

A IC ₅₀ < 10 nM

B 10 nM < IC ₅₀ < 100 nM

C 100 nM < IC ₅₀ < 1,000 nM

D 1,000 nM < IC ₅₀ < 10,000 nM

E IC ₅₀ > 10,000 nM

Table 2.

Connection number PIKK Family PI3K family ATR mTOR PI3Kα PI3Kα (E542A) PI3Kβ PI3Kδ PI3Kγ 1 E C C C D C D 2 E C C C D C D 3 E D D D D C D 4 E D D D D C D 5 E D D D D C D 6 E B C C C B C 7 E C D D D C E 8 E C C C C B D 9 E C D D E D E 10 E D D D D C D

The results indicate that the synthesized compounds exhibit pharmacologically significant activity and selectivity against ATM and DNA-PK, while demonstrating no significant activity against ATR and mTOR, as well as several PI3K kinases, such as PI3Kα, PI3Kβ, PI3Kδ, and PI3Kγ. The compounds exhibit inhibitory activity against ATM and DNA-PK, simultaneously with high reliability, as determined in a direct in vitro assay using a generally accepted and validated method.

Claims (24)

1. The use of a pharmaceutical composition having inhibitory activity against ATM and DNA-PK, containing a therapeutically effective amount of a compound defined by formula (I) or its pharmaceutically acceptable salt, where - Y may be -S- or -SO ₂ -, then Z ₁ and Z ₂ are independently selected and represent -CH- or N; or Y may represent -O-, then one of Z ₁ and Z ₂ represents N and the other represents -CH-; - M ₁ and M ₂ are hydrogen or M ₁ and M ₂ together form =O (keto group); - n is 0 or 1; - R ₁ is -OLN(R _a ,R _b ); - R ₂ is C _1-3 alkyl, C ₃ cycloalkyl or phenyl; - R ₃ is a halogen; - R ₄ is hydrogen or C _1-2 alkyl; - each R _a and R _b independently represents hydrogen or C _1-3 alkyl; - L represents ethylene or n-propylene, and a pharmaceutically acceptable excipient for the manufacture of a medicinal product for the treatment of an oncological disease treatable with a dual inhibitor of ATM and DNA-PK. 2. The use according to claim 1, wherein the oncological disease is acute myeloid leukemia, acute lymphoblastic leukemia, acute megakaryocytic leukemia, promyelocytic leukemia, erythroleukemia, lymphoblastic T-cell leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, hairy cell leukemia, chronic neutrophilic leukemia, plasmacytoma, immunoblastic large cell leukemia, mantle cell leukemia, multiple myeloma, malignant lymphoma, diffuse large B-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, lymphoblastic T-cell lymphoma, Burkitt's lymphoma, follicular lymphoma, brain cancer (e.g. astrocytoma, glioma, glioblastoma, medulloblastoma, ependymoma), bladder cancer, breast cancer, central nervous system tumors, cervical cancer, rectal cancer, colon cancer, endometrial cancer, esophageal cancer, gastrointestinal stromal tumor, gastric cancer, head and neck cancer, oral cavity cancer, hepatocellular carcinoma, cholangiocarcinoma, metastatic liver disease, Merkel cell carcinoma, lung cancer, melanoma, mesothelioma, nasopharyngeal cancer, neuroblastoma, osteosarcoma, ovarian cancer, pancreatic cancer, prostate cancer, kidney cancer, salivary gland cancer, sarcoma, testicular cancer, urothelial cancer, vulvar cancer, and Wilms' tumor. 3. Use of the composition specified in paragraph 1 for the manufacture of a medicinal product for the treatment of an oncological disease treatable with a dual inhibitor of ATM and DNA-PK in a patient undergoing radiotherapy. 4. The use according to claim 3, wherein the composition is administered to the patient simultaneously with radiotherapy. 5. The use according to claim 3, wherein the composition is administered to the patient before radiotherapy. 6. The use according to claim 3, wherein the composition is administered to the patient after radiotherapy. 7. Use of the composition specified in paragraph 1 for the manufacture of a medicinal product for the treatment of an oncological disease treatable with a dual inhibitor of ATM and DNA-PK in a patient who receives an antitumor agent selected from the following list: cisplatin, oxaliplatin, carboplatin, valrubicin, idarucin, calicheamicin, olaparib, rucaparib, niraparib, veliparib, talazoparib, ipilimumab, ofatumumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab. 8. The use according to claim 7, wherein the composition is administered to the patient simultaneously with an antitumor agent. 9. The use according to claim 7, wherein the composition is administered to the patient before the antitumor agent. 10. The use according to claim 7, wherein the composition is administered to the patient after the antitumor agent.