EA045102B1 - APPLICATION OF 1-[4-BROMO-5-[1-ETHYL-7-(METHYLAMINO)-2-OXO-1,2-DIHYDRO-1,6-NAPHYRIDIN-3-YL]-2-FLUOROPHENYL]-3-PHENYLUREA AND ANALOGUES FOR THE TREATMENT OF CANCER ASSOCIATED WITH GENETIC DISORDERS IN THE PLATELET GROWTH FACTOR ALPHA RECEPTOR - Google Patents

Description

Field of technology to which the invention relates

The present invention relates to the use of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl]-3 -phenylurea or 1 -(5-(7-amino-1-ethyl-2-oxo-1,2dihydro-1,6-naphthyridin-3-yl)-4-bromo-2-fluorophenyl)-3-phenylurea in the treatment types of cancer. In particular, the present invention is directed to methods of inhibiting PDGFR kinases and methods of treating cancers and disorders associated with inhibition of PDGFR kinases, including lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors tract (GIST), malignant peripheral nerve sheath sarcoma, intimal sarcomas, hypereosinophilic syndrome, acute myeloid leukemia associated with eosinophilia, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, or lymphoblastic T-cell lymphoma.

Prerequisites for creating the invention

Oncogenic genomic alterations of PDGFRa kinase or overexpression of PDGFRa kinase have been shown to cause human cancers.

Missense mutations of the PDGFRa kinase have been shown to cause a subset of GIST tumors. PDGFRa mutations are oncogenic in approximately 8-10% of GIST cases (Corless, Modern Pathology 2014;27:S1-16). The predominant PDGFRa mutation is D842V in exon 18, although other mutations in exon 18 have also been reported, including D846Y, N848K, and Y849K, and insertion-deletion mutations (INDEL) in exon 18, including RD841-842KI, DI842-843-IM and HDSN845-848P. In addition, rare mutations in exons 12 and 14 of PDGFRa have also been reported (Corless et al, J Clinical Oncology 2005;23:5357-64).

Deletion mutations in exon 18 of PDGFRa AD842-H845 and AI843-D846 have been described in GIST (Lasota et al, Laboratory Investigation 2004;84:874-83).

Amplification or mutation of PDGRFa has been described in human malignant peripheral nerve sheath tumor (MPNST) tissues (Holtkamp et al, Carcinogenesis 2006;27:664-71).

PDGFRa amplification has been described in multiple skin lesions in undifferentiated pleomorphic sarcoma (Osio et al, J. Cutan Pathol 2017;44:477-79) and in intimal sarcoma (Zhao et al, Genes Chromosomes and Cancer 2002;34:48- 57;Dewaele et al, Cancer Res 2010;70:730414).

PDGFRa amplification has been associated with a subset of patients with lung cancer. 4q12, containing the PDGFRa gene locus, is amplified in 3-7% of lung adenocarcinomas and 8-10% of lung squamous cell carcinomas (Ramos et al, Cancer Biol Ther. 2009;8:2042-50; Heist et al., J Thorac Oncol 2012;7:924-33).

Mutations in the IDH protein produce a novel oncometabolite, 2-hydroxyglutarate, which interacts with iron hydroxylases, including the TET family of 5'-methylcytosine hydroxylases. TET enzymes catalyze the key step in removing DNA methylation. Flavahan et al. demonstrated that human gliomas involving mutant IDH are characterized by increased levels of methylation at the DNA binding sites of cohesin and CCTC-binding factor (CTCF), impairing the binding of this methylation-sensitive insulator protein (Flavahan et al., Nature 2016;529:110). Reduced CTCF binding is associated with loss of insulation between topological domains and aberrant gene activation. In particular, loss of CTCF at the domain boundary allows the constitutive enhancer to aberrantly interact with the PDGFRA receptor tyrosine kinase gene, which is a major glioma oncogene. Thus, cancers involving mutated IDH may be prone to mediate oncogenic events through activation and overexpression of wild-type PDGFRa.

PDGFRa amplification is common in pediatric and adult high-grade astrocytomas and provides the opportunity to identify a group with poor prognostic features in glioblastoma involving mutant IDH1. PDGFRa amplification was frequently observed in pediatric (29.3%) and adult (20.9%) tumors. PDGFRa amplification has been reported to increase with increasing grade and is particularly associated with a poorer prognosis in de novo GBM involving mutant IDH1 (Phillips et al., Brain Pathology, 2013;23:565-73).

It was shown that the presence of an intragenic deletion rearrangement of PDGFRa exon 8.9 was detected in PDGFRa-amplified gliomas at the PDGFRa locus. This intragenic deletion is common and is present in 40% of glioblastoma multiforme (GBM) cases presenting with PDGFRa amplification. Tumors with this rearrangement demonstrate histological

- 1 045102 features of oligodendroglioma, and intragenic deletion of exon 8.9 of PDGFRa shows persistently increased tyrosine kinase activity (Ozawa et al., Genes and Development 2010; 24: 2205-18).

The FIP1L1-PDGFRA fusion protein is oncogenic in a subgroup of patients suffering from hypereosinophilic syndrome (Elling et al., Blood 2011;117;2935). The FIP1L1-PDGFRa fusion has also been identified in eosinophilia-associated acute myeloid leukemia and lymphoblastic T-cell lymphoma (Metzgeroth et al., Leukemia 2007; 21: 1183-88).

Thus, mutations, deletions, rearrangements and amplifications of the PDGFRa gene are associated with a number of solid and hematological cancers. Given the complex function of the PDGFRa gene and the potential utility of PDGFRa inhibitors in the treatment of various solid and hematological cancers, there is a need for inhibitors with good therapeutic properties.

Brief description of the invention

One aspect of the present invention relates to a method of treating or preventing tumor growth or tumor progression mediated by PDGFR kinase, comprising administering to a patient in need thereof an effective amount of 1-[4-bromo-5-[1-ethyl-7-(methylamino)- 2-oxo1,2-dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl]-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of inhibiting PDGFR kinase comprising administering to a patient in need thereof an effective amount of 1-[4-bromo-5-[1ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1 ,6-naphthyridin-3-yl]-2-fluorophenyl]-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of inhibiting PDGFR kinase or treating tumor growth or tumor progression mediated by PDGFR kinase. The method involves administering to a patient in need 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2dihydro-1,6-naphthyridin-3-yl]-2- fluorophenyl]-3-phenylurea or a pharmaceutically acceptable salt thereof, alone or in combination with other anti-cancer therapeutic agents, anti-cancer biological agents, immune checkpoint inhibitors or chemotherapeutic agents.

Another aspect of the present invention provides a method of treating glioblastoma comprising administering to a patient in need thereof an effective amount of 1-[4-bromo-5-[1-ethyl-7(methylamino)-2-oxo-1,2-dihydro-1 ,6-naphthyridin-3-yl]-2-fluorophenyl]-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of treating PDGFRa-mediated gastrointestinal stromal tumors comprising administering to a patient in need thereof an effective amount of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2- oxo-1,2-dihydro1,6-naphthyridin-3-yl]-2-fluorophenyl]-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of treating or preventing tumor growth or tumor progression mediated by PDGFR kinase, comprising administering to a patient in need thereof an effective amount of 1-(5-(7-amino-1-ethyl-2-oxo-1, 2dihydro-1,6-naphthyridin-3-yl)-4-bromo-2-fluorophenyl)-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of inhibiting PDGFR kinase, comprising administering to a patient in need thereof an effective amount of 1-(5-(7-amino-1ethyl-2-oxo-1,2-dihydro-1,6-naphthyridine-3 -yl)-4-bromo-2-fluorophenyl)-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of inhibiting PDGFR kinase or treating tumor growth or tumor progression mediated by PDGFR kinase. The method involves administering 1-(5-(7-amino-1-ethyl-2-oxo-1,2-dihydro-1,6naphthyridin-3-yl)-4-bromo-2-fluorophenyl) to a patient in need. -3-phenylurea or a pharmaceutically acceptable salt thereof, alone or in combination with other anticancer therapeutic agents, anticancer biological agents, immune checkpoint inhibitors or chemotherapeutic agents.

Another aspect of the present invention provides a method of treating glioblastoma comprising administering to a patient in need thereof an effective amount of 1-(5-(7-amino-1-ethyl-2oxo-1,2-dihydro-1,6-naphthyridine-3- yl)-4-bromo-2-fluorophenyl)-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to a method of treating PDGFRa-mediated gastrointestinal stromal tumors comprising administering to a patient in need thereof an effective amount of 1-(5-(7-amino-1-ethyl-2-oxo-1,2- dihydro-1,6-naphthyridin3-yl)-4-bromo-2-fluorophenyl)-3-phenylurea or a pharmaceutically acceptable salt thereof.

Another aspect of the present invention relates to the in vivo biosynthetic production of 1-(5-(7amino-1-ethyl-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl)-4-bromo-2-fluorophenyl )-3-phenylurea (compound B) after oral administration of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1,6naphthyridine- 3-yl]-2-fluorophenyl]-3-phenylurea (compound A).

The present invention further provides methods for inhibiting PDGFR kinases and treating cancers and disorders associated with inhibition of PDGFR kinases, including lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant sarcoma peripheral nerve sheaths, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma.

The present invention also provides methods for inhibiting PDGFRa kinase, oncogenic PDGFRa missense mutations, oncogenic PDGFRa deletion mutations, oncogenic PDGFRa gene rearrangements resulting in PDGFRa fusion proteins, or oncogenic amplification of the PDGFRa gene.

The present invention also provides methods of using 1-[4-bromo-5-[1-ethyl-7(methylamino)-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl ]-3-phenylurea or 1-(5-(7amino-1-ethyl-2-oxo-1,2-dihydro-1,6-naphthyridin-3 -yl)-4-bromo-2-fluorophenyl)-3 - phenylurea.

Brief description of graphic materials

In fig. 1A-1C show MRI images of the brain of a patient with a glioblastoma tumor, showing PDGFRa amplification. In fig. 1A shows an MRI image of the patient's brain at baseline. In fig. 1B shows evidence of tumor reduction after cycle 9. FIG. Figure 1C shows an MRI image of the same brain after cycle 12.

Detailed Description of the Invention

It was found that 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1,6-naphthyridin-3yl]-2-fluorophenyl]-3- phenylurea (compound A) and 1-(5-(7-amino-1-ethyl-2-oxo-1,2-dihydro-1,6naphthyridin-3-yl)-4-bromo-2-fluorophenyl)-3- Phenylurea (compound B) unexpectedly inhibits wild-type PDGFR kinases and their oncogenic protein forms. The present invention provides a method of treating cancer by inhibiting tumor growth or tumor progression mediated by oncogenic PDGFRa kinase, comprising administering to a patient in need thereof an effective amount of 1 -[4-bromo-5-[ 1 -ethyl-7-(methylamino)- 2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl]-2fluorophenyl]-3-phenylurea, 1 -(5 -(7-amino-1-ethyl-2-oxo-1,2- dihydro-1,6-naphthyridin-3-yl)-4-bromo-2fluorophenyl)-3-phenylurea or a pharmaceutically acceptable salt thereof.

Definition

Compounds A and B as used herein refer to 1-[4-bromo-5-[1-ethyl-7(methylamino)-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl ]-2-fluorophenyl]-3-phenylurea and 1-(5-(7amino-1-ethyl-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl)-4-bromo-2- fluorophenyl)-3-phenylurea. The present invention also provides pharmaceutically acceptable salts, tautomers, hydrates and solvates of compounds A and B. The structures of compounds A and B are presented below:

- [4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl]-3 phenylurea (compound A )

1-(5-(7-amino-1-ethyl-2-oxo-1,2-dihydro-1,6-naphthyirdin-3-yl)-4-bromo-2-fluorophenyl)-3phenylurea (compound B).

Methods for preparing Compound A and Compound B are disclosed in US Pat. No. 8,461,179 B1, the contents of which are incorporated herein by reference. Details of the present invention are set forth below in the accompanying description. Although methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention, exemplary methods and materials are described herein. Other features, objects and advantages of the present invention will be apparent from the description and claims. In the specification and the accompanying claims, the singular form also includes the plural unless the context clearly requires otherwise. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which the present invention belongs.

- 3 045102

Throughout this disclosure, reference is made to various patents, patent applications, and publications. The disclosures of these patents, patent applications, and publications are incorporated herein by reference in their entirety to more fully describe the state of the art known to those skilled in the art as of the date of this disclosure. This invention will govern if there is any inconsistency between patents, patent applications and publications and this invention.

For convenience, certain terms used in the description, examples, and claims are collected herein. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as commonly understood by one skilled in the art to which the present invention belongs. The original definition provided for a group or term given in this disclosure applies to that group or term throughout this disclosure, alone or as part of another group, unless otherwise indicated.

A pharmaceutically acceptable carrier, diluent or excipient includes, without limitation, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, coloring agent, flavor enhancer, surfactant, wetting agent, dispersant an agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the United States Food and Drug Administration and is acceptable for use in humans or domestic animals. The pharmaceutically acceptable salt includes both acid addition salts and base addition salts.

Pharmaceutically acceptable acid addition salt refers to those salts that retain the biological effectiveness and properties of free bases that are not biologically or otherwise undesirable and that are prepared with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid acid, phosphoric acid and the like, 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 кислота, муравьиная кислота, фумаровая кислота, галактаровая кислота, гентизиновая кислота, глюкогептоновая кислота, глюконовая кислота, глюкуроновая кислота, глутаминовая кислота, глутаровая кислота, 2оксо-глутаровая кислота, глицерофосфорная кислота, гликолевая кислота, гиппуровая кислота, изомасляная кислота, молочная кислота, лактобионовая acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucinic 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, pyruvic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p- toluenesulfonic acid, trifluoroacetic acid, undecenoic acid, etc.

A pharmaceutical composition refers to a composition of a compound of the present invention and a medium conventional in the art for delivering a biologically active compound to mammals, such as humans. Such a medium includes all pharmaceutically acceptable carriers, diluents or excipients therefor.

Subjects or patients in need of treatment with a compound of the present invention, or patients in need of PDGFRa inhibition, include patients with diseases and/or conditions that can be treated with the compounds of the present invention to achieve a beneficial therapeutic result. Favorable outcome includes objective response, improved progression-free survival, improved survival, prolonged stable disease phase, and/or decreased severity of symptoms or delayed onset of symptoms. For example, a patient in need of treatment is suffering from tumor growth or tumor progression; the patient suffers from, without limitation, adenocarcinoma of the lung, squamous cell carcinoma of the lung, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant sarcoma of the peripheral nerve sheaths, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma.

As used herein, an effective amount (or pharmaceutically effective amount) of a compound disclosed herein is an amount that results in a favorable clinical outcome for the condition being treated with the compound compared to no treatment. The amount of compound or compounds administered will depend on the extent, severity and type of disease or condition, the amount of therapy required, and the release characteristics of the pharmaceutical composition. It will also depend on the health, size, weight, age, gender and drug tolerance of the subject. Typically, the compound is administered for a sufficient period of time to achieve the desired therapeutic effect.

The terms treat, treat, and treat are intended to include the full range of interventions for a patient with cancer for the purpose of preventing the growth of a tumor from which the patient is suffering and/or preventing the progression of a tumor under a given treatment, such as administering an active compound to alleviate, slow, or reverse one or more symptoms and delay the progression of cancer, even if the cancer is not actually eliminated. Treatment may promote recovery, improvement, or at least partial relief of the disorder.

Cancer, as defined herein, refers to a neoplasm that has the ability to invade surrounding tissue, metastasize (spread to other organs), and which may ultimately lead to the death of the patient if left untreated. The cancer may be a solid tumor or hematologic malignancy.

The term tumor as used herein refers to a tumor formation. The term can refer to benign (generally harmless) or malignant (cancerous) growths. Malignant growth may originate from a solid organ or bone marrow. The latter is often referred to as hemoblastosis.

Tumor growth, as defined herein, refers to tumor growth driven by genomic alterations of the PDGFRa kinase.

Tumor progression, as defined herein, refers to the growth of an existing PDGFRa-dependent tumor, where such growth of an existing tumor lesion is due to additional PDGFRa kinase genomic alterations that are resistant to treatment.

One aspect of the present invention relates to a method of treating or preventing tumor growth or tumor progression mediated by PDGFR kinase, comprising administering to a patient in need thereof an effective amount of 1-[4-bromo-5-[1-ethyl-7-(methylamino)- 2-oxo1,2-dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl]-3-phenylurea (compound A) or a pharmaceutically acceptable salt thereof.

In one embodiment, Compound A or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein tumor growth or tumor progression is due to overexpression of PDGFRa kinase, oncogenic PDGFRa missense mutations, oncogenic PDGFRa deletion mutations, oncogenic PDGFRa gene rearrangements resulting in PDGFRa fusions proteins, intragenic deletions of PDGFRa in frame and/or oncogenic amplification of the PDGFRa gene. In one embodiment, tumor growth or tumor progression is caused by overexpression of PDGFRa kinase. In another embodiment, tumor growth or tumor progression is caused by oncogenic missense mutations of PDGFRa. In another embodiment, tumor growth or tumor progression is caused by oncogenic PDGFRa deletion mutations. In another embodiment, tumor growth or tumor progression is caused by oncogenic rearrangements of the PDGFRa genes resulting in PDGFRa fusion proteins. In another embodiment, tumor growth or tumor progression is caused by intragenic in-frame deletions of PDGFRa. In another embodiment, tumor growth or tumor progression is caused by oncogenic amplification of the PDGFRa gene.

In another embodiment, Compound A or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein tumor growth or tumor progression is caused by mutant PDGFRa with the D842V mutation, mutant PDGFRa with the V561D mutation, deletion mutations in exon 18 of PDGFRa, including mutant PDGFRa with the 842 deletion -845, intragenic deletion in exon 8.9 of PDGFRa, in-frame fusions to PDGFRa, including FIP1L1-PDGFRa, or amplification of PDGFRa.

In another embodiment, Compound A or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein the cancer is lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant sarcoma peripheral nerve sheaths, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma. In one embodiment, the cancer is a glioblastoma. In another embodiment, the cancer is a gastrointestinal stromal tumor.

In another embodiment, Compound A or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, either as a single agent or in combination with other anticancer therapeutics, anticancer biologics, immune checkpoint inhibitors, or chemotherapeutic agents.

Another aspect of the present invention relates to a method of treating or preventing tumor growth or tumor progression mediated by PDGFR kinase, comprising administering to a patient in need thereof an effective amount of 1-(5-(7-amino-1-ethyl-2-oxo-1, 2dihydro-1,6-naphthyridin-3-yl)-4-bromo-2-fluorophenyl)-3-phenylurea (compound B) or a pharmaceutically acceptable salt thereof.

In one embodiment, Compound B or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein tumor growth or tumor progression is due to overexpression of PDGFRa kinase, oncogenic PDGFRa missense mutations, oncogenic PDGFRa deletion mutations, oncogenic PDGFRa gene rearrangements resulting in PDGFRa fusions proteins, intragenic deletions of PDGFRa in frame and/or oncogenic amplification of the PDGFRa gene. In one embodiment, tumor growth or tumor progression is caused by overexpression of PDGFRa kinase. In another embodiment, tumor growth or tumor progression is caused by oncogenic missense mutations of PDGFRa. In another embodiment, tumor growth or tumor progression is caused by oncogenic PDGFRa deletion mutations. In another embodiment, tumor growth or tumor progression is caused by oncogenic rearrangements of the PDGFRa genes resulting in PDGFRa fusion proteins. In another embodiment, tumor growth or tumor progression is caused by intragenic in-frame deletions of PDGFRa. In another embodiment, tumor growth or tumor progression is caused by oncogenic amplification of the PDGFRa gene.

In another embodiment, Compound B or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein tumor growth or tumor progression is caused by mutant PDGFRa with the D842V mutation, mutant PDGFRa with the V561D mutation, deletion mutations in exon 18 of PDGFRa, including mutant PDGFRa with the 842 deletion -845, intragenic deletion in exon 8.9 of PDGFRa, in-frame fusions to PDGFRa, including FIP1L1-PDGFRa, or amplification of PDGFRa.

In another embodiment, Compound B or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, wherein the cancer is lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant sarcoma peripheral nerve sheaths, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma. In one embodiment, the cancer is a glioblastoma. In another embodiment, the cancer is a gastrointestinal stromal tumor. In another embodiment, Compound B or a pharmaceutically acceptable salt thereof is administered to a patient suffering from cancer, either as a single agent or in combination with other anticancer therapeutic agents, anticancer biologics, immune checkpoint inhibitors, or chemotherapeutic agents.

Pharmaceutical compositions and methods of treatment

In addition, it should be noted that the present invention is directed to methods of treatment involving the administration of a compound of the present invention or a pharmaceutical composition containing such a compound. The pharmaceutical composition or preparation described herein can be used in accordance with the present invention for the treatment of various types of cancer, including lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant peripheral nerve sheath sarcoma, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma.

The compounds used in the treatment methods of the present invention, as well as pharmaceutical compositions containing them, may suitably be administered alone or as part of a patient's treatment protocol or treatment regimen that involves the administration or use of other useful compounds (which are further described elsewhere herein). document).

In some embodiments, the present invention relates to a method of using a pharmaceutical composition comprising Compound A or B and a pharmaceutically acceptable carrier providing one or more additional therapeutic agents. Additional therapeutic agents include, but are not limited to, cytotoxic agent, cisplatin, doxorubicin, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, lonafarnib, tipifarnib, 4-((5-((4-( 3chlorophenyl)-3-oxopiperazin-1 -yl)methyl)-1 H-imidazol-1 -yl)methyl)benzonitrile hydrochloride, (R)-1 ((1H-imidazol-5-yl)methyl)-3-benzyl- 4-(thiophen-2-ylsulfonyl)-2,3,4,5-tetrahydro-1H-benzodiazepine-7- 6 045102 carbonitrile, cetuximab, imatinib, interferon alpha-2b, pegylated interferon alpha-2b, combinations with aromatase, gemcitabine , uramustine, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylene thiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, leucovorin, oxaliplatin, pen tostatin, vinblastine, vincristine , vindesine, bleomycin, dactinomycin, daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17a-ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone , megestrol acetate, methylprednisolone, methyltestosterone, prednisolone , triamcinolone, chlorotrianisene, 17a-hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide acetate, flutamide, toremifene citrate, goserelin acetate, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, vinorelbine, ana strozole, letrozole, capecitabine, raloxifene, droloxafine, hexamethylmelamine , bevacizumab, trastuzumab, tositumomab, bortezomib, ibritumomab tiuxetan, arsenic trioxide, sodium porfimer, cetuximab, thiotepa, altretamine, fulvestrant, exemestane, rituximab, alemtuzumab, dexamethasone, bicalutamide, chlorambucil and valrubicin.

In other embodiments, the present invention relates to a method of using a pharmaceutical composition comprising Compound A or B and a pharmaceutically acceptable carrier providing one or more additional therapeutic agents. Additional therapeutic agents may include, but are not limited to, an AKT inhibitor, an alkylating agent, a polytransretinoic acid, an antiandrogen, azacitidine, a BCL2 inhibitor, a BCL-XL inhibitor, a BCR-ABL inhibitor, a BTK inhibitor, a BTK/LCK/LYN inhibitor, a CDK1/2/4/inhibitor. 6/7/9, CDK4/6 inhibitor, CDK9 inhibitor, CBP/p300 inhibitor, EGFR inhibitor, endothelin receptor antagonist, ERK inhibitor, farnesyltransferase inhibitor, FLT3 inhibitor, glucocorticoid receptor agonist, HDM2 inhibitor, histone deacetylase inhibitor, IKKe inhibitor, immunomodulatory drug agent (IMiD), ingenol, ITK inhibitor, JAK1/JAK2/JAK3/TYK2 inhibitor, MEK inhibitor such as but not limited to trametinib, selumetinib and cobimetinib, midostaurin, MTOR inhibitor, PI3 kinase inhibitor, dual PI3/MTOR kinase inhibitor, proteasome inhibitor , protein kinase C agonist, SUV39H1 inhibitor, TRAIL, VEGFR2 inhibitor, Wnt/βcatenin signaling pathway inhibitor, decitabine and anti-CD20 monoclonal antibody.

In other embodiments, the present invention provides a pharmaceutical composition comprising Compound A or B and a pharmaceutically acceptable carrier containing therapeutically effective amounts of one or more additional therapeutic agents, wherein said additional therapeutic agents are immune checkpoint inhibitors, and which are selected from the group, consisting of CTLA4 inhibitors such as, but not limited to, ipilimumab and tremelimumab; PD1 inhibitors such as, but not limited to, pembrolizumab and nivolumab; PDL1 inhibitors such as, but not limited to, atezolizumab (formerly MPDL3280A), MEDI4736, avelumab, PDR001; 4 1BB or 4 1BB ligand inhibitors such as, without limitation, urelumab and PF-05082566; or OX40 ligand agonists such as without limitation MEDI6469; GITR inhibitors such as, but not limited to, TRX518; CD27 inhibitors such as, but not limited to, varlilumab; TNFRSF25 or TL1A inhibitors; CD40 agonists such as, but not limited to, CP-870893; HVEM or LIGHT or LTA or BTLA or CD160 inhibitors; LAG3 inhibitors such as, but not limited to, BMS-986016; TIM3 inhibitors; Siglec inhibitors; ICOS agonists or ICOS ligand; B7 H3 inhibitors such as, but not limited to, MGA271; B7 H4 inhibitors; VISTA inhibitors; HHLA2 or TMIGD2 inhibitors; butyrophilin inhibitors, including BTNL2 inhibitors; CD244 or CD48 inhibitors; inhibitors of members of the TIGIT and PVR families; KTR inhibitors such as, without limitation, lirilumab; ILT and LIR inhibitors; inhibitors of NKG2D and NKG2A, such as but not limited to IPH2201; MICA and MICB inhibitors; CD244 inhibitors; CSF1R inhibitors such as, but not limited to, emactuzumab, cabiralizumab, pexidartinib, ARRY382, BLZ945; IDO inhibitors such as, but not limited to, INCB024360; TGFe inhibitors such as, but not limited to, galunisertib; adenosine or CD39 or CD73 inhibitors; inhibitors of CXCR4 or CXCL12, such as, but not limited to, ulocuplumab and (3 S,6S,9S,12R, 17R,20S,23 S,26S,29S,34aS)-N-((S)-1-amino-5-guanidino- 1-oxopentan-2-yl)-26,29bis(4-aminobutyl)-17-((S)-2-((S)-2-((S)-2-(4-fluorobenzamido)-5-guanidinopentanamido )-5-guanidinopentanamido)-3-(naphthalene-2-yl)propanamido)-6-(3-guanidinopropyl)-3,20-bis(4-hydroxybenzyl)1,4,7,10,18,21,24 ,27,30-nonaoxo-9,23-bis(3-ureidopropyl)triacontahydro-1H,16H-pyrrolo[2,1р][1,2]dithia[5,8,11,14,17,20,23, 26,29]nonaazacyclodotriacontane-12-carboxamide BKT140; phosphatidylserine inhibitors such as, but not limited to, bavituximab; inhibitors of SIRPA or CD47, such as but not limited to CC-90002; VEGF inhibitors such as, but not limited to, bevacizumab; and neuropilin inhibitors, such as but not limited to MNRP1685A.

When using pharmaceutical compositions based on the compounds described herein, pharmaceutically acceptable carriers can be either solid or liquid. Solid forms include powders, tablets, dispersible granules, capsules, coated preparations and soup

- 7 045102 positions. Powders and tablets may contain from about 5 to about 95% active ingredient. Suitable solid carriers are known in the art, for example magnesium carbonate, magnesium stearate, talc, sugar or lactose. Tablets, powders, coated formulations and capsules may be used in solid dosage forms suitable for oral administration. Examples of pharmaceutically acceptable carriers and methods for preparing various compositions can be found in A. Gennaro (ed.), Remington's Pharmaceutical Sciences, 18th Edition, (1990), Mack Publishing Co., Easton, Pa., which is incorporated herein by reference in its entirety. completeness.

Preparations in liquid form include solutions, suspensions and emulsions. For example, aqueous solutions or solutions of water and propylene glycol for parenteral injection, or the addition of sweeteners and opacifiers for oral solutions, suspensions and emulsions. Preparations in liquid form may also include solutions for intranasal administration.

Liquid, in particular injectable, compositions can, for example, be prepared by dissolving, dispersing, etc. For example, the disclosed compound is dissolved or mixed with a pharmaceutically acceptable solvent, such as, for example, water, saline, aqueous dextrose, glycerol, ethanol, and the like, thereby obtaining an isotonic injection solution or suspension. Proteins such as albumin, chylomicron particles or serum proteins can be used to solubilize the disclosed compounds.

Parenteral administration by injection is typically used for subcutaneous, intramuscular or intravenous injections and infusions. Injectable preparations can be prepared in conventional forms, either as liquid solutions or as suspensions or solid forms suitable for dissolution in liquid prior to injection.

Aerosol preparations suitable for inhalation can also be used. These preparations may include solutions and solids in powder form, which may be in combination with a pharmaceutically acceptable carrier such as an inert pressurized gas such as nitrogen.

It also contemplates the use of solid form preparations that are intended to be converted shortly before use into liquid form preparations for either oral or parenteral administration. Such liquid forms include solutions, suspensions and emulsions.

Types of combination therapy

As noted previously, the compounds described herein can be used alone or in combination with other agents. For example, the compounds can be administered together with an anticancer therapeutic agent, an anticancer biological agent, an immune checkpoint inhibitor, or a chemotherapeutic agent. In another embodiment, compound A or B may be the only one used or used alone with another agent. The agent can be administered together or sequentially with a compound described herein in combination therapy.

Combination therapy can be achieved by administering two or more agents, each formulated and administered separately, or by administering two or more agents in a single formulation. Other combinations are also covered by combination therapy. For example, two agents can be formulated together and administered together with a separate formulation containing a third agent. Although two or more agents in combination therapy can be administered simultaneously, this is not required. For example, administration of the first agent (or combination of agents) may precede administration of the second agent (or combination of agents) by minutes, hours, days or weeks. Thus, two or more agents can be administered within a few minutes of each other, or within 1, 2, 3, 6, 9, 12, 15, 18 or 24 hours of each other, or within 1, 2, 3 , 4, 5, 6, 7, 8, 9, 10, 12, 14 days from each other, or within 2, 3, 4, 5, 6, 7, 8, 9 or weeks from each other. In some cases, even longer intervals are possible. Although in many cases it is desirable for two or more agents used in combination therapy to be present in the patient's body at the same time, this is not necessary.

Combination therapy may also include two or more administrations of one or more agents, used in combination, using different sequences of the constituent agents. For example, if agent X and agent Y are used in combination, they may be administered sequentially in any combination one or more times, for example in the order X-Y-X, X-X-Y, Y-X-Y, YY-X, X-X-Y-Y, etc.

In one embodiment, Compound A or B is administered to a patient in need of treatment in combination with a therapeutic agent selected from: a cytotoxic agent, cisplatin, doxorubicin, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, lonafarnib, tipifarnib, 4-((5((4-(3-chlorophenyl)-3-oxopiperazin-1 -yl)methyl)-1 H-imidazol-1 -yl)methyl)benzonitrile hydrochloride, (R)-1 -((1H-imidazol-5-yl)methyl)-3-benzyl-4-(thiophen-2-ylsulfonyl)-2,3,4,5-tetrahydro-1H-benzodiazepine-7- carbonitrile, cetuximab, imatinib, interferon alpha-2b, pegylated interferon alpha-2b, combinations with aromatase, gemcitabine, uramustine, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine,

- 8 045102 lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, leucovorin, oxaliplatin, pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin -C, L-asparaginase, teniposide 17a-ethinylestradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, 17αhydroxyprogesterone, aminogl utetimide, estramustine, medroxyprogesterone acetate, leuprolide acetate, flutamide, toremifene citrate , goserelin acetate, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, vinorelbine, anastrozole, letrozole, capecitabine, raloxifene, droloxafine, hexamethylmelamine, bevacizumab, trastuzumab, tositumomab, bortezomib, and britumomab tiuxetan, arsenic trioxide, sodium porfimer, cetuximab, thiotepa, altretamine, fulvestrant, exemestane, rituximab, alemtuzumab, dexamethasone, bicalutamide, chlorambucil and valrubicin.

In one embodiment, Compound A or B is administered to a patient in need of treatment in combination with immune checkpoint inhibitors selected from CTLA4 inhibitors such as, but not limited to, ipilimumab and tremelimumab; PD1 inhibitors such as, but not limited to, pembrolizumab and nivolumab; PDL1 inhibitors such as, but not limited to, atezolizumab (formerly MPDL3280A), MEDI4736, avelumab, PDR001; 4 1BB or 4 1BB ligand inhibitors such as, without limitation, urelumab and PF-05082566; or OX40 ligand agonists such as without limitation MEDI6469; GITR inhibitors such as, but not limited to, TRX518; CD27 inhibitors such as, but not limited to, varlilumab; TNFRSF25 or TL1A inhibitors; CD40 ligand agonists such as, but not limited to, CP870893; HVEM or LIGHT or LTA or BTLA or CD160 inhibitors; LAG3 inhibitors such as, but not limited to, BMS-986016; TIM3 inhibitors; Siglec inhibitors; ICOS inhibitors or ICOS ligand; B7 H3 inhibitors such as, but not limited to, MGA271; B7 H4 inhibitors; VISTA inhibitors; HHLA2 or TMIGD2 inhibitors; butyrophilin inhibitors, including BTNL2 inhibitors; CD244 or CD48 inhibitors; inhibitors of members of the TIGIT and PVR families; KIR inhibitors such as, but not limited to, lirilumab; ILT and LIR inhibitors; inhibitors of NKG2D and NKG2A, such as but not limited to IPH2201; MICA and MICB inhibitors; CD244 inhibitors; CSF1R inhibitors such as, but not limited to, emactuzumab, cabiralizumab, pexidartinib, ARRY382 and BLZ945; IDO inhibitors such as, but not limited to, INCB024360; TGFe inhibitors such as, but not limited to, galunisertib; adenosine or CD39 or CD73 inhibitors; inhibitors of CXCR4 or CXCL12, such as, but not limited to, ulocuplumab and (3S,6S,9S,12R,17R,20S,23S,26S,29S,34aS)-N-((S)-1-amino-5-guanidino-1oxopentane- 2-yl)-26,29-bis(4-aminobutyl)-17-((S)-2-((S)-2-((S)-2-(4-fluorobenzamido)-5-guanidinopentanamido)- 5-guanidinopentanamido)-3-(naphthalene-2-yl)propanamido)-6-(3-guanidinopropyl)-3,20bis(4-hydroxybenzyl)-1,4,7,10,18,21,24,27, 30-nonaoxo-9,23-bis(3-ureidopropyl)triacontahydro1H,16H-pyrrolo[2,1-p][1,2]dithia[5,8,11,14,17,20,23,26,29 ]nonaazacyclodotriacontin-12-carboxamide BKT140; phosphatidylserine inhibitors such as, but not limited to, bavituximab; inhibitors of SIRPA or CD47, such as but not limited to CC-90002; VEGF inhibitors such as, but not limited to, bevacizumab; or neuropilin inhibitors, such as but not limited to MNRP1685A.

In another embodiment of the present invention, additional therapeutic agents may be used in combination with Compound A or B. Such agents include, but are not limited to, an AKT inhibitor, an alkylating agent, polytransretinoic acid, an antiandrogen, azacitidine, a BCL2 inhibitor, a BCL-XL inhibitor, a BCR- ABL, BTK inhibitor, BTK/LCK/LYN inhibitor, CDK1/2/4/6/7/9 inhibitor, CDK4/6 inhibitor, CDK9 inhibitor, CBP/p300 inhibitor, EGFR inhibitor, endothelin receptor antagonist, ERK inhibitor, farnesyl transferase inhibitor , FLT3 inhibitor, glucocorticoid receptor agonist, HDM2 inhibitor, histone deacetylase inhibitor, IKKe inhibitor, immunomodulatory drug (IMiD), ingenol, radiation exposure, ITK inhibitor, JAK1/JAK2/JAK3/TYK2 inhibitor, MEK inhibitor such as but not limited to trametinib, selumetinib and cobimetinib, midostaurin, MTOR inhibitor, PI3 kinase inhibitor, dual PI3/MTOR kinase inhibitor, proteasome inhibitor, protein kinase C agonist, SUV39H1 inhibitor, TRAIL, VEGFR2 inhibitor, Wnt/β-catenin signaling pathway inhibitor, decitabine and monoclonal antibody to CD20.

Dose

In some embodiments, where Compound A or B is used in combination with another agent for a treatment protocol, the composition may be administered together or in a dual regimen where the two therapeutic agents are dosed and administered separately. If Compound A or B and the additional agent are dosed separately, the typical dose administered to a subject in need of treatment is typically from about 5 mg per day to about 5000 mg per day, and in other embodiments from about 50 mg per day to approximately 1000 mg per day. Other doses may range from about 10 mmol to about 250 mmol per day, from about 20 mmol to about 70 mmol per day, or even from about 30 mmol to about

- 9 045 102 mmol per day.

The amount and frequency of administration of the compounds of the present invention and/or pharmaceutically acceptable salts thereof are adjusted in accordance with the judgment of the attending physician, taking into account factors such as the age, condition and weight of the patient, as well as the severity of the symptoms to be treated. Amounts of effective doses of the disclosed compounds, when used to obtain the indicated effects, range from about 0.5 mg to about 5000 mg of the disclosed compound needed to treat the condition. Compositions for in vivo or in vitro use may contain about 0.5, 5, 20, 50, 75, 100, 150, 250, 500, 750, 1000, 1250, 2500, 3500 or 5000 mg of the disclosed compound, or in the range of one quantities to another quantity in the list of doses. A typical recommended daily dosage regimen for oral administration may range from about 1 mg/day to about 500 mg/day, or from 1 mg/day to 200 mg/day in a single dose or two to four divided doses. In one embodiment, the typical daily dose of the regimen is 150 mg.

The compounds of the present invention, with or without additional agents described herein, can be administered by any suitable route. The compound can be administered orally (eg, with food) in capsules, suspensions, tablets, pills, dragees, liquids, gels, syrups, suspensions, and the like. Methods for encapsulating the compositions (such as coating with hard gelatin or cyclodextran) are known in the art ( Baker, et al., Controlled Release of Biological Active Agents, John Wiley and Sons, 1986, which is incorporated herein by reference in its entirety). The compounds can be administered to a subject in combination with an acceptable pharmaceutical carrier as part of a pharmaceutical composition. The composition of the pharmaceutical composition will vary depending on the chosen route of administration. Suitable pharmaceutical carriers may contain inert ingredients that do not react with the compound. The carriers are biologically compatible, i.e. non-toxic, non-inflammatory, non-immunogenic and free of other adverse reactions at the injection site.

Exemplary pharmaceutical compositions are tablets and gelatin capsules containing a compound of the present invention and a pharmaceutically acceptable carrier such as a) a diluent such as purified water, triglyceride oils such as hydrogenated or partially hydrogenated vegetable oil or mixtures thereof, corn oil, olive oil , sunflower oil, safflower oil, types of fish oils such as EPA or DHA or their esters or triglycerides or mixtures thereof, omega-3 fatty acids or their derivatives, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin , glucose and/or glycine; b) a lubricant, for example silicon dioxide, talc, stearic acid, magnesium or calcium salts thereof, sodium oleic acid, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and/or polyethylene glycol; also for tablets; c) a binder, for example magnesium aluminosilicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, magnesium carbonate, natural sugars such as glucose or beta-lactose, corn sugar, natural and synthetic gums such as acacia, tragacanth or sodium alginate, waxes and/or polyvinylpyrrolidone, if necessary; d) a disintegrant, for example starch, agar, methylcellulose, bentonite, xanthan gum, alginic acid or its sodium salt or foaming mixtures; f) absorbent, coloring matter, flavoring and sweetener; f) an emulsifier or dispersant such as Tween 80, Labrazole, HPMC, DOSS, Caproyl 909, Labrafac, Labrafil, Peceol, Transcutol, Capmule MSM, Capmule PG-12, Captex 355, Gelucir, Vitamin E TGPS or other suitable emulsifier; and/or g) an agent that improves the absorption of the compound, such as cyclodextrin, hydroxypropylcyclodextrin, PEG400, PEG200.

When formulated as a fixed dose, such combination products utilize the compounds of the present invention within the dosage range described herein or known to those skilled in the art.

Since the compounds of the present invention (compounds A and B) are intended for use in pharmaceutical compositions, those skilled in the art will appreciate that they can be provided in substantially pure forms, e.g., at least 60% pure, at least 75% pure. % pure, at least 85% pure and at least 98% pure (w/w). The pharmaceutical preparation can be obtained in a standard dosage form. In such form, the preparation is divided into unit doses of suitable size containing suitable amounts of compounds A or B, for example an effective amount to achieve the desired purpose described herein.

Section 1 - Important structural comparisons with biological activity in WO/2008/034008 and WO/2013/184119

WO/2008/034008 describes various kinases that cause or contribute to the pathogenesis of various proliferative diseases, these kinases include the families BRaf, CRaf, Abl, KDR(VEGFR2), EGFR/HER1, HER2, HER3, c-MET, FLT-3, PDGFR-α, PDGFR-β, p38, with KIT, JAK2. The disclosure of this PCT application shows selective inhibition of Braf and CRaf kinases with

- 10 045102 using analogues of compounds A and B described in this document. Concurrently, WO/2013/184119 describes the inhibition of mutant c-KIT by compounds A and B. However, WO/2013/184119 also discloses that c-KIT and PDGFRa mutations are mutually exclusive in GIST. This is because most GISTs have primary activating mutations in genes encoding the closely related RTKs c-KIT (75-80% of GISTs) or PDGFRa (8% of non-mutated c-KIT GISTs), in a mutually exclusive manner.

Herein, the inexorable mutual exclusivity between c-KIT and PDGFRa mutations in patients with GIST is consistent with the finding that Compounds A and B can treat both patient populations. In fact, unexpectedly, Compounds A and B, which are known to inhibit mutant c-KIT, also inhibit wild-type PDGFR kinases and oncogenic mutated PDGFR kinases, oncogenic PDGFRa kinase fusion protein forms, and PDGFRa amplification-induced cancers, contrary to previous findings. disclosures WO/2008/034008 and WO/2013/184119. Experimental data described below further supports this finding. An immediate application of this discovery is the treatment of subpopulations of cancer patients who exhibit resistant forms of the cancers described herein and which are driven by PDGFR.

Examples

Biological data

Compounds A and B were found to surprisingly inhibit wild-type PDGFR kinases and oncogenic mutated PDGFR kinases, oncogenic PDGFRa kinase fusion proteins, and cancers due to PDGFRa mutations or amplifications thereof. This unexpected discovery was investigated using biochemical assays, cellular assays and in vivo clinical evaluation in cancer patients.

The present invention is further illustrated by the following examples, which should not be construed as limiting the present invention in the scope or spirit of the specific procedures described herein. It should be understood that the examples are provided to illustrate certain embodiments and that the scope of the present invention is not intended to be limited thereby. It should also be understood that, in practice, there may be various other embodiments, modifications and equivalents thereof that may be suggested to those skilled in the art without departing from the spirit of the present invention and/or the scope of the appended claims.

Example 1: Inhibition of the enzymatic activity of wild-type PDGFRa.

Biochemical analysis of PDGFRa (GenBank accession number: NP_006197)

PDGFRa kinase activity was determined spectroscopically using a coupled pyruvate kinase/lactate dehydrogenase assay that continuously monitors ATP hydrolysis-dependent NADH oxidation (eg, Schindler et al. Science (2000) 289: 1938-1942, which is incorporated herein by reference in its entirety). Assays were performed in 384-well plates (100 μl final volume) using 4.8 nM PDGFRA (DeCode Biostructures, Bainbridge Island, WA), 5 units pyruvate kinase, 7 units lactate dehydrogenase, 1 mM phosphoenolpyruvate, 0.28 mM NADH, 2.5 mg/ml PolyEY and 0.5 mM ATP in assay buffer (90 mM Tris, pH 7.5, 18 mM MgCl ₂ , 1 mM DTT and 0.2% octyl glucoside). PDGFRA inhibition was measured after addition of serially diluted test compound (final assay concentration 1% DMSO). The decrease in absorbance at 340 nm was monitored continuously for 6 h at 30°C on a multimode microplate reader (BioTek, Winooski, Vermont). Reaction rates were calculated using a time frame of 1-2 hours. Reaction rates at each compound concentration were converted to percentage inhibition using controls (i.e., reaction without test compound and reaction with known inhibitor) and IC ₅₀ values were calculated by fitting the sigmoidal curve with four parameters on the data using Prism (GraphPad, San Diego, CA).

- 11 045102

PDGFRa protein sequence (residues 550-1089 with N-terminal GST tag;

SEQ ID NO: 1 in Genbank)

MEFIHHHHHHHMAPILGYWKIKGL.VQPTRLI.LEYLF.EKYEEHLYERDEGDKWRNKKFE. LGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYGVS RIAYSKDEETLKVDFLSKLPEMLKMFEDRLCHKTYLN(jDHVTHPDFMLYDA.LDVVLY NiDPMCLDAFPKLVCFKKRIEAIPQIDKVTKSSKYIAWPLQGWQATFGGGDHPPKSDLVP RHNQTSLY KKAGFEGDRTMKQKPRYE1RWRVIES1SPDGHEY1YVDPMQLPYDSRWEFP RDGLVLGRVLGSGAFGKVVEGTAYGLSRSQPVMKVAVKMLKPTARSSEKQALMSELKI MTHLGPHLNIVNLLGACTKSGPIYIITEYCFYGDLWYLHKNRDSFLSHHPEKPKKELDIF GLNPADESTRSY VJLSFENNGDYMDMKQADTEQYVPMLERKEVSKYSDIQRSLYDRPA SYKKKSMLDSEVKNLLSDDNSEGLTIJLDLLSFTYQVARGMEFLASKNCVHRDLAARNV LLAQGKIVKICDFGLARDIMHDSNYVSKGSTFLPVKWMAPESIFDNLYTTLSDVWSYGI LLWEIFSLCKjTPYPGMMVDSITYNKIKSGYRM AKPDH ATSEVYEIMVKCWNSEPEKRP SFYHLSEIVXMT.XGQYKKSYEKffiLDFLKSDin’AVARMRVDSDNAYTGVTYKNEEDKL KDWEGGLDEQRLSADSGYIIPLPDIDPVPEEEDLGKRNRHSSQTSEESAIETGSSSSTFIKR EDETIED 1DMMDDIGIDS SDL VTDSFL

Compound A inhibited the enzymatic activity of recombinant wild-type PDGFRa with an IC ₅₀ value of 12 nM. Compound B inhibited the enzymatic activity of recombinant wild-type PDGFRa with an IC ₅₀ value of 6 nM.

Example 2. Inhibition of enzymatic activity of PDGFRa with the D842V mutation.

Biochemical analysis of PDGFRa D842V (GenBank accession number: NP_006197)

PDGFRA D842V kinase activity was determined spectroscopically using a coupled pyruvate kinase/lactate dehydrogenase assay that continuously monitors ATP hydrolysis-dependent NADH oxidation (e.g., Schindler et al. Science (2000) 289: 1938-1942, which is incorporated herein by references in their entirety). Assays were performed in 384-well plates (100 μl final volume) using 3 nM PDGFRA D842V (Invitrogen, Carlsbad, CA), 5 units pyruvate kinase, 7 units lactate dehydrogenase, 1 mM phosphoenolpyruvate, 0.28 mM NADH, 2.5 mg /ml PolyEY and 0.5 mM ATP in assay buffer (90 mM Tris, pH 7.5, 18 mM MgCl ₂ , 1 mM DTT and 0.2% octyl glucoside). PDGFRA inhibition by D842V was measured after addition of serially diluted test compound (final assay concentration 1% DMSO). The decrease in absorbance at 340 nm was monitored continuously for 6 hours at 30°C on a multimode microplate reader (BioTek, Winooski, Vermont). Reaction rates were calculated using a time frame of 2-3 hours. Reaction rates at each compound concentration were converted to percentage inhibition using controls (i.e., reaction without test compound and reaction with known inhibitor) and IC ₅₀ values were calculated by fitting the sigmoidal curve with four parameters on the data using Prism (GraphPad, San Diego, CA).

Protein sequence PDGFRa D842V (residues 550-1089 with N-terminal HIS-GST tag;

SEQ ID NO: 2 in Genbank)

MAPILGYW’KIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKFELGLEFPNLPYYI DGDVKLTQSMA11RYIADKHNM LGGCPK ERAEISMLEGA VLDIRYGVSRIA YSKDFETLK VDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLYMDPMCLDAFPKL VCFKKRI EAIPQroKYLKSSKYIAWPLQGWQATFGGGDHPPKSDLVPRHNQTSLYKKAG FEGDRTMKQKPRYEIRWRVIESISPDGHEYIYVDPMQLPYDSRWEFPRDGLVLGRVLGS GAFGKVYEGTAYGLSRSQPVMKVAVKMLKPTARSSEKQALMSELKIMTHHLGPHLNrVN LLGACTKSGPIY I]TEYCFYGDLVNYLHKNRDStT.SHHPEKPKKELDIFGLNPADESTRSY YTLSFENNGDYMDMKQADTTQYATMLERKEVSKYSDIQRSLYDRPASYKKKSMIXSEV KNLL SDDN SEGLTLLDLLSFT YQ VARGMEFLA SKNC VHRDLAARN VLL AQGK1VK1CDF GLARVIMHDSNYVSKGST FLPVKWMAPESIFDNLYTTLSDVWSYG1LLWEIFSLGGTPYP GMMVDSTFYNKIKSGYRMAKPDHATSEVYEIMVKCWNSEPEKRPSFYHLSEIVENLLPG QYKKSYEKIHLDFLKSDHPAVARMRVDSDNAYIGVTYKNEEDKLKDWEGGLDEQRLS ADSGYIIPLPDIDPWEEEDLGKRNREISSQTSEESAI ETGSSSSTFIKREDETEDIDMMDDI GIDSSDLVEDSFL

Compound A inhibited the enzymatic activity of recombinant PDGFRa with the D842V mutation with an IC ₅₀ value of 42 nM. Compound B inhibited the enzymatic activity of recombinant PDGFRa with the D842V mutation with an IC ₅₀ value of 20 nM.

Example 3 Inhibition of wild-type PDGFRp enzymatic activity.

Biochemical analysis of PDGFRB (GenBank accession number: NP_002600)

PDGFRP kinase activity was determined spectroscopically using a coupled pyruvate kinase/lactate dehydrogenase assay that continuously monitors ATP hydrolysis-dependent NADH oxidation (e.g., Schindler et al. Science (2000) 289: 1938-1942, which is incorporated herein by reference in its entirety). Assays were performed in 384-well plates (100 µl final volume) using 9 nM PDGFRB (DeCode Bio structures, Bainbridge Island, WA), 5 units pyruvate kinase, 7 units lactate dehydrogenase, 1 mM phosphoenolpyruvate, 0.28 mM NADH, 2.5 mg /ml PolyEY and 0.5 mM ATP in assay buffer (90 mM Tris, pH 7.5, 18 mM MgCl ₂ , 1 mM DTT and 0.2% octyl glucoside). PDGFRB inhibition was measured after addition of serially diluted test compound (final assay concentration 1% DMSO). The decrease in absorbance at 340 nm was monitored continuously for 6 h at 30°C on a multimode microplate reader (BioTek, Winooski, Vermont). The reaction rate was calculated using a time frame of 2-3 hours. The reaction rate at

- 12045102 each compound concentration was converted to percent inhibition using controls (i.e., a reaction without the test compound and a reaction with a known inhibitor) and IC ₅₀ values were calculated by fitting a four-parameter sigmoidal curve to the data using Prism (GraphPad, San -Diego, California).

PDGFRP protein sequence (residues 557-1106 with N-terminal HIS-GST tag: SEQ ID NO: 3 in Genbank)

MEHHIH LI Π Π Π IM APILGYWKIKGLVQPTRLLLEYLEEK YEEHLYERDEGDK W RNK KFE LGLEFPNI,PYY^IDGDVKLTQSMAnRYIADKHNNn.GGCPKERAErSML.EGA\TDIRYGVS RLAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPD FMLYDALDVVLY MDPMCLDARPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQAJTGGGDHPPKSDLVP RHNQTSLYKKAGFEGDRTMQKKPRYEIRWKVIESVSSDGHEYIYVDPMQLPYDSTWEL PRDQL VLGRTLGSGAFGQVVEATAHGLSHSQATMKVAVKMLKS TARS SEKQ ALM SEL KIMSHLGPIR,NVVNLLGACTKGGPIYIITEYCRYGDLVDYLHRNKHTFLQHHSDKRRPP SAELYSNALPVGLPLPSHVSLTGESDGGYMDMSKDESVDYVPMLDMKGDVKYADIESS N YM APYDN YVPSAP ERTCR A FUN ESPVLS YMDL VGF S YQ V ANGMEELASKNC VHRDL AARNVLTCEGKLVKICDFGLARDTMRDSNYISKGSTFLPLKWMAPESIFNSLYTTLSDVW SFGILL WEIFTLGGIPYPELPMNEQF YN AIKRGYRMAQP AHASDE1YEIM QKC WEEKFEI RPPFSQLVLLLERLLGEGYKKKYQQVDEEFLRSDHPA1LRSQARLPGFHGLRSPLDTSSV LYTAVQPN EGDKDYIIPLPDPKPEVADEGPLEGSPSLASSTLNEVNTSSTISCDSPLEPQDE PEPEPQLELQVEPEPELEQLPDSGCPAPRAEAEDSFL

Compound A inhibited the enzymatic activity of recombinant wild-type PDGFRP with an IC ₅₀ value of 9 nM. Compound B inhibited the enzymatic activity of recombinant wild-type PDGFRp with an IC ₅₀ value of 5 nM.

Example 4 Inhibition of the increase in the amount of PDGFRa with the D842V mutation expressed in Ba/F3 cells.

Cell culture BaF3 PDGFRa D842V

BaF3 cells were transfected with a construct encoding PDGFRa D842V and selected independently of IL-3. Briefly, cells were grown in RPMI 1640 medium supplemented with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 1 U/ml penicillin G, 1 μg/ml streptomycin, and 0.29 mg/ml L-glutamine at 37 °C, 5% CO ₂ , 95% humidity.

BaF3 cell proliferation assays with PDGFRa D842V

Serial dilutions of the test compound were dispensed into a 96-well black clear bottom plate (Corning, Corning, NY). Ten thousand cells were added per well to 200 μl of prepared culture medium. The plates were incubated for 67 hours at 37°C, 5% CO ₂ , 95% humidity. At the end of the incubation period, 40 μl of a 440 μM solution of resazurin (Sigma, St. Louis, MO) in PBS was added to each well and the plates were incubated for an additional 5 h at 37°C, 5% CO ₂ , 95% humidity. The plates were read on a Synergy2 reader (Biotek, Winooski, Vermont) using excitation at 540 nm and emission at 600 nm. Data were analyzed using Prism software (GraphPad, San Diego, CA) to calculate _IC50 values.

Compound A inhibited the proliferation of BaF3 cells with PDGFRa with the D842V mutation with an IC ₅₀ value of 36 nM. Compound B inhibited the proliferation of BaF3 cells with PDGFRa with the D842V mutation with an IC50 value of 42 nM.

Example 5: Inhibition of phosphorylation of PDGFRa with D842V mutation expressed in BaF3 cells.

Cell culture BaF3 PDGFRa D842V

BaF3 cells were transfected with a construct encoding PDGFRa D842V and selected independently of IL-3. Briefly, cells were grown in RPMI 1640 medium supplemented with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 1 U/ml penicillin G, 1 μg/ml streptomycin, and 0.29 mg/ml L-glutamine at 37 °C, 5% CO ₂ , 95% humidity.

Western blotting BaF3 PDGFRa D842V

Two million cells per well suspended in serum-free RPMI 1640 were added to a 24-well tissue culture plate. Serial dilutions of the test compound were added to plates containing cells, and the plates were incubated for 4 hours at 37°C, 5% CO ₂ , 95% humidity. Cells were washed with PBS and then lysed. Cell lysates were separated by SDS-PAGE and transferred to PVDF. Phospho-PDGFRa (Tyr754) was detected using an antibody from Cell Signaling Technology (Beverly, MA), ECL Plus developing reagent (GE Healthcare, Piscataway, NJ), and a Molecular Devices Storm 840 fluorescent phosphor imager. fluorescence mode. Blots were stripped and probed for total PDGFRa using an antibody from Cell Signaling Technology (Beverly, MA). IC ₅₀ values were calculated using Prism software (GraphPad, San Diego, CA).

Compound A inhibited phosphorylation of PDGFRa with D842V mutation expressed in BaF3 cells with an IC ₅₀ value of 24 nM. Compound B inhibited the phosphorylation of PDGFRa with the D842V mutation expressed in BaF3 cells with an IC ₅₀ value of 26 nM.

- 13 045102

Example 6. Inhibition of phosphorylation of PDGFRa with the V561D mutation expressed in CHO cells.

Chinese hamster ovary (CHO) cells were transiently transfected with a PDGFRA cDNA construct with the V561D mutation cloned into plasmid pcDNA3.1 (Invitrogen, Carlsbad, CA). Twenty-four hours after transfection, cells were treated with various concentrations of the compound for 90 minutes. Protein lysates from cells were prepared and immunoprecipitated using an anti-PDGFRA antibody (SC-20, Santa Cruz Biotechnology, Santa Cruz, CA) followed by sequential immunoblotting for phosphotyrosine using a monoclonal antibody (PY-20, BD Transduction Labs, Sparks, MD) or generic PDGFRa (SC-20, Santa Cruz Biotechnology, Santa Cruz, CA). Densitometry was performed to quantify drug effect using Photoshop 5.1 software, with phospho-PDGFRa levels normalized to total protein. Experimental densitometry results were analyzed using Calcusyn 2.1 software (Biosoft, Cambridge, UK) to mathematically determine IC ₅₀ values.

Compound A inhibited phosphorylation of the V561D mutant PDGFRa expressed in CHO cells with an IC ₅₀ value of 25 nM.

Example 7. Inhibition of phosphorylation of PDGFRa with deletion mutation 842-845 in exon 18 expressed in CHO cells.

Chinese hamster ovary (CHO) cells were transiently transfected with a PDGFRA cDNA construct with the AD842-H845 mutation cloned into plasmid pcDNA3.1 (Invitrogen, Carlsbad, CA). 24 h after transfection, cells were treated with various concentrations of the compound for 90 min. Protein lysates from cells were prepared and immunoprecipitated using an anti-PDGFRA antibody (SC-20, Santa Cruz Biotechnology, Santa Cruz, CA) followed by sequential immunoblotting for phosphotyrosine using a monoclonal antibody (PY-20, BD Transduction Labs, Sparks, MD) or generic PDGFRa (SC-20, Santa Cruz Biotechnology, Santa Cruz, CA). Densitometry was performed to quantify drug effect using Photoshop 5.1 software, with phospho-PDGFRA levels normalized to total protein. Experimental densitometry results were analyzed using Calcusyn 2.1 software (Biosoft, Cambridge, UK) to mathematically determine IC ₅₀ values.

Compound A inhibited phosphorylation of PDGFRa with the 842-845 deletion mutation in exon 18 expressed in CHO cells with an IC ₅₀ value of 77 nM.

Example 8 Inhibition of the increase in FIP1L1-PDGFRa fusion in EOL-1 cells.

EOL-1 cell culture (FIP1L1/PDGFRa fusion)

EOL-1 cells were grown in RPMI 1640 medium supplemented with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 1 U/ml penicillin G, 1 μg/ml streptomycin, and 0.29 mg/ml L-glutamine at 37°C, 5% CO ₂ , 95% humidity.

EOL-1 Cell Proliferation Assays

Serial dilutions of the test compound were dispensed into a 96-well black clear bottom plate (Corning, Corning, NY). Ten thousand cells were added per well to 200 μl of prepared culture medium. The plates were incubated for 67 hours at 37°C, 5% CO ₂ , 95% humidity. At the end of the incubation period, 40 μl of a 440 μM solution of resazurin (Sigma, St. Louis, MO) in PBS was added to each well and the plates were incubated for an additional 5 h at 37°C, 5% CO ₂ , 95% humidity. The plates were read on a Synergy2 reader (Biotek, Winooski, Vermont) using excitation at 540 nm and emission at 600 nm. Data were analyzed using Prism software (GraphPad, San Diego, CA) to calculate _IC50 values.

Compound A inhibited the increase in FIP1L1-PDGFRa fusion in EOL-1 cells with an IC ₅₀ value of 0.029 nM. Compound B inhibited the increase in FIP1L1-PDGFRa fusion in EOL-1 cells with an IC ₅₀ value of 0.0 1 8 nM.

Example 9 Inhibition of phosphorylation of the FIP1L1-PDGFRa fusion in EOL-1 cells.

EOL-1 cell culture (FIP1L1/PDGFRa fusion)

EOL-1 cells were grown in RPMI 1640 medium supplemented with 10% characterized fetal bovine serum (Invitrogen, Carlsbad, CA), 1 U/ml penicillin G, 1 μg/ml streptomycin, and 0.29 mg/ml L-glutamine at 37°C, 5% CO ₂ , 95% humidity.

Western blot analysis of EOL-1

Two million cells per well suspended in serum-free RPMI 1640 were added to a 24-well tissue culture plate. Serial dilutions of the test compound were added to plates containing cells, and the plates were incubated for 4 hours at 37°C, 5% CO ₂ , 95% humidity. Cells were washed with PBS and then lysed. Cell lysates were separated with

- 14 045102 using SDS-PAGE and transferred to PVDF. Phospho-PDGFRa (Tyr754) was detected using an antibody from Cell Signaling Technology (Beverly, MA), ECL Plus developing reagent (GE Healthcare, Piscataway, NJ), and a Molecular Devices Storm 840 fluorescent phosphor imager. fluorescence mode. Blots were stripped and probed for total PDGFRa using an antibody from Cell Signaling Technology (Beverly, MA). IC ₅₀ values were calculated using Prism software (GraphPad, San Diego, CA).

Compound A inhibited phosphorylation of the FIP1L1-PDGFRa fusion in EOL-1 cells with an IC ₅₀ value of 0.12 nM. Compound B inhibited phosphorylation of the FIP1L1-PDGFRa fusion in EOL-1 cells with an _IC50 value of <0.1 nM.

Example 10 Treatment of human cancer patients with PDGFRa with the D842V mutation.

Clinical Study Protocol DCC-2618-01-001 An open-label, multicenter, Phase 1 study of Compound A to evaluate safety, tolerability, and pharmacokinetics in patients with advanced tumors represents the first human study of Compound A (ClinicalTrials.gov Identifier: NCT02571036). The objectives of this dose escalation study are to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary antitumor activity of Compound A. The study drug was administered orally either once or twice daily at escalating doses ranging from 20 mg. BID up to 200 mg BID. Preliminary antitumor activity was measured by CT scan according to RECIST 1.1 every second cycle (every 56 days). Pharmacodynamic effects were measured as reduction in mutation allele frequency (MAF) in cell-free plasma DNA (cf) and analyzed using Guardant 360 version 2.9 or version 2.10 (Guardant Health, Redwood City, CA), a next-generation sequencing platform for 73 genes.

All patients had disease that progressed with standard treatment and progressed rapidly without treatment. The study included three patients with PDGFRa mutant gastrointestinal stromal tumors (GISTs). The D842V PDGFRa mutation was identified in each patient by tumor biopsy. Based on non-clinical data and available pharmacokinetic data from study DCC-2618-01-001, dose levels >50 mg BID (daily dose equivalent to 100 mg) were sufficient to result in tumor control, i.e. growth arrest in such advanced sarcomas from PDGFRa-dependent tumors with the D842V mutation in patients suffering from GIST. Of the 3 evaluable patients, 2 enrolled at or above the study effective dose level (150 mg QD and 100 mg BID). Another patient was enrolled at 30 mg BID and progress was noted after 2 cycles of treatment for 28 days. The 100 mg BID patient was on cycle 11 (>40 weeks) and continued to benefit from treatment. The most recent tumor evaluation confirmed stable disease according to RECIST 1.1. Tumor assessments throughout the study showed some tumor reduction (5-10%), including the most recent assessment after cycle 9 (36 weeks). The patient who was treated at 150 mg QD was on cycle 6 (>20 weeks) with stable disease according to RECIST and had some tumor shrinkage. 2 patients received 1 and 3 pretreatments with tyrosine kinase inhibitors, respectively.

To date, cfDNA surveillance data regarding the incidence of the PDGFRa D842V allele mutation in plasma are only available for the 100 mg BID patient. The D842V mutation in PDGFRa was not detected by cfDNA at baseline, but a frequency of 0.59% was detected at day 1 of cycle 3 (8 weeks) after treatment. Although failure to detect the D842V mutation at baseline may limit the ability to interpret the data, the fact that the mutation found in tumor tissue is undetectable i.e. below the detection limit at 2 consecutive assay points (cycle 5 day 1 (16 weeks) and cycle 7 day 1 (24 weeks)) strongly supports suppression of this D842V mutation in PDGFRa as a result of treatment of human cancer patients with the compound A.

Example 11 Treatment of a patient suffering from human glioblastoma with PDGFRa amplification.

Clinical Study Protocol DCC-2618-01-001 An open-label, multicenter, Phase 1 study of Compound A to evaluate safety, tolerability, and pharmacokinetics in patients with advanced tumors represents the first human study of Compound A (ClinicalTrials.gov Identifier: NCT02571036). The objectives of this dose escalation study are to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary antitumor activity of Compound A. The study drug was administered orally either once or twice daily at escalating doses ranging from 20 mg. BID up to 200 mg BID. Preliminary antitumor activity was measured by CT scan according to RANO (Revised Assessment in NeuroOncology) criteria every cycle after every 3 ^cycles (every 56 or 84 days). Pharmacodynamic effects were measured as a decrease in the number of circulating tumor cells (CTC). Target

- 15 045102 new blood was enriched for STS in an OncoQuick tube. The CTC layer was incubated with an adenovirus that replicates and expresses GFP in cells with high levels of telomerase (Oncolys BioPharma Inc.). Cells were then incubated with fluorescently labeled antibodies, fixed, and stained with DAPI. Cells positive for DAPI, GFP, PDGFRa, and GFAP fluorescence were counted as circulating glioblastoma tumor cells using a BioTek Cytation 5 imaging device. Gliofibrillary acidic protein (GFAP) is uniquely associated with glial cells.

All patients had disease that progressed with standard treatment and progressed rapidly without treatment. One patient with PDGFRa-amplified glioblastoma (GBM; 6x amplified, 12 copies) was enrolled in the study at 20 mg BID. The patient was initially treated with combination radiochemotherapy followed by temozoloid alone and progressed after 3 months. The GBM patient was in cycle 19 (>17 months in the study) and continued to benefit from treatment. Since tumor evaluation was performed after cycle 12 (48 weeks), the patient had a partial response according to RANO criteria. In fig. 1 shows an MRI image at baseline (FIG. 1A) and after cycle 12 (FIG. 1C). In fig. Figure 1B provides further evidence of tumor reduction after cycle 9.

The significance of PDGFRa amplification was assessed in high-grade astrocytomas (HGA) in children and adults, including glioblastomas. A large study of primary human tissue suggests a significant prevalence of PDGFRa-induced HGA and indicates that PDGFRa amplification increases with grade and is associated with a poorer prognosis in de novo GBM involving mutant IDH1 (Philips et al., Brain Pathol. (2013) 23 (5):565-73, which is incorporated herein by reference in its entirety). Dunn et al. provide further evidence that PDGFRa amplification is a driver of genomic changes in GBM (Dunn et al., Genes Dev. (2012) 26(8):756-84). Based on these results, the pharmacodynamic effect, measured as the reduction in CTC observed in the GBM patient after treatment with Compound A, strongly confirms that the partial response observed in the GBM patient is a result of treatment of the PDGFRa-amplified tumor with the compound A. Double positive STS (PDGFRa+/GFAP+) were first measured during cycle 7 (28 weeks) at a rate of 2.22 STS/ml. The frequency decreased in cycles 13 (52 weeks) and 17 (68 weeks) to 1.11 and 0.58 CTC/ml, respectively.

Example 12 Compound B is produced by biological synthesis after oral administration of Compound A.

Clinical Study Protocol DCC-2618-01-001 An open-label, multicenter, Phase 1 study of Compound A to evaluate safety, tolerability, and pharmacokinetics in patients with advanced tumors represents the first human study of Compound A (ClinicalTrials.gov Identifier: NCT02571036). The objectives of this dose escalation study are to evaluate the safety, tolerability, pharmacokinetics (PK), pharmacodynamics (PD), and preliminary antitumor activity of Compound A. The study drug was administered orally either once or twice daily at escalating doses ranging from 20 mg. BID up to 200 mg BID. Oral administration of Compound A to patients results in systemic exposure to Compound A and biotransformation of Compound A to Compound B via N-demethylation in vivo. For pharmacokinetic (PK) analysis, blood samples were obtained on cycle 1, day 15 immediately before the morning dose of Compound A and at 0.5, 1, 2, 4, 6, 8, and 10-12 hours post-dose. Compound A and its active metabolite, compound B, were analyzed using a validated bioanalytical method. Phoenix WinNonlin version 6.3 was used for plasma concentration-time analysis to calculate standard non-compartmental PK parameters. All PK calculations were performed using nominal sample collection times.

As an example, administration of Compound A to a cohort of patients at doses of 150 mg twice daily or 150 mg once daily resulted in stable exposure to Compound A on day 15 of cycle 1, as well as Compound B, as indicated in the table below.

An oral dose of 150 mg of Compound A was administered BID (twice daily) to a cohort of 5 patients for 15 days, exposed to Compound A with a mean Cmax of 1,500 ng/mL and mean area under the curve (AUC) of 11,400 ng/h /ml. This administration over 15 days resulted in biotransformation to Compound B with an average Cmax=1.520 ng/ml and average AUC=15.100 ng*h/ml. An oral dose of 150 mg of Compound A was administered QD (once daily) to a cohort of 4 patients for 15 days, exposed to Compound A with a mean Cmax of 861 ng/mL and mean area under the curve (AUC) of 8.070 ng* h/ml. This dosing over 15 days resulted in biotransformation to Compound B with an average Cmax=794 ng/ml and average AUC=8,600 ng*h/ml.

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Table

Oral dose of compound A Compound A Stach (ng/ml) Compound A AUC12 h (ng*h/ml) Connection B Stach (ng/ml) Connection B AUC12 4 (ng*h/ml) 150 mg BID 1,500 11,400 1,520 15,100 150 mg QD 861 8,070 794 8,600

Equivalents

One skilled in the art will recognize or be able to ascertain, using no more than routine experimentation, numerous equivalents to the specific embodiments specifically described herein. Such equivalents are intended to be included within the scope of the claims below.

Claims (37)

1. A method of treating or preventing tumor growth or tumor progression mediated by PDGFR kinase in a patient in need thereof, comprising administering to the patient 150 mg once daily or twice daily of a compound represented by the formula or a pharmaceutically acceptable salt thereof. 2. The method according to claim 1, wherein tumor growth or tumor progression is caused by one or more of overexpression of PDGFRa kinase, oncogenic missense mutations of PDGFRa, oncogenic deletion mutations of PDGFRa, oncogenic rearrangements of PDGFRa genes leading to PDGFRa fusion proteins, intragenic deletions in PDGFRa in-frame or oncogenic amplification of the PDGFRa gene. 3. The method according to claim 1 or 2, wherein tumor growth or tumor progression is caused by overexpression of PDGFRa kinase. 4. The method according to claim 1 or 2, wherein tumor growth or tumor progression is caused by oncogenic PDGFRa missense mutations or oncogenic PDGFRa deletion mutations. 5. The method according to claim 1 or 2, wherein tumor growth or tumor progression is caused by oncogenic rearrangements of the PDGFRa genes leading to PDGFRa fusion proteins, or intragenic deletions in PDGFRa in frame. 6. The method according to claim 1 or 2, wherein tumor growth or tumor progression is caused by oncogenic amplification of the PDGFRa gene. 7. Method according to any of pi. 1-6, where the tumor is a lung adenocarcinoma, squamous cell lung cancer, glioblastoma, childhood glioma, astrocytomas, sarcomas, gastrointestinal stromal tumors, malignant peripheral nerve sheath sarcoma, intimal sarcomas, hypereosinophilic syndrome, idiopathic hypereosinophilic syndrome, chronic eosinophilic leukemia, acute myeloid leukemia associated with eosinophilia, or lymphoblastic T-cell lymphoma. 8. Method according to any one of claims 1 to 7, where the tumor is a glioblastoma. 9. Method according to any one of claims 1 to 7, wherein the tumor is a stromal tumor of the gastrointestinal tract. 10. Method according to any of pi. 1-9, where 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1,6naphthyridin-3-yl]-2-fluorophenyl]-3 β-phenylurea or a pharmaceutically acceptable salt thereof is administered as a single agent or in combination with other anticancer therapeutic agents, anticancer biological agents, immune checkpoint inhibitors or chemotherapeutic agents. I. The method according to claim 10, where the therapeutic agent is selected from a cytotoxic agent, cisplatin, doxorubicin, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, lonafarnib, tipifarnib, 4- ((5-((4-(3-chlorophenyl)-3-oxopiperazin-1-yl)methyl)-1H-imidazol-1-yl)methyl)benzonitrile hydrochloride, (R)-l-((lH-imidazol- 5-yl)methyl)-3-benzyl-4-(τthiophene-2-ylcysulfonyl)-2,3,4,5-τeτpahydro-lHbenzodiazepine-7-carbonitrile, cetuximab, imatinib, interferon alpha-2b, pegylated interferon alpha- 2b, combinations with aromatase, gemcitabine, uramustine, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocine, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6thio guanine, fludarabine phosphate, leucovorin, oxaliplatin, pentostatin, vinblastine, vincristine, - 17045102 vindesine, bleomycin, dactinomycin, daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17a-ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanol it is propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, 17a-hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide acetate, flutamide, toremifene citrate, goserelin acetate, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, lion amizole, vinorelbine, anastrozole, letrozole, capecitabine, raloxifene, droloxafine, hexamethylmelamine, bevacizumab, trastuzumab, tositumomab, bortezomib, ibritumomab tiuxetan, arsenic trioxide, sodium porfimer, cetuximab, thiotepa, altretamine, fulvestrant, exemestane, rituximab, alemtuzumab, dexamethasone, bicalutamide, chlorambucil or val rubicin. 12. The method of claim 10, wherein the immune checkpoint inhibitor is selected from CTLA4 inhibitors, ipilimumab and tremelimumab; PD1 inhibitors, pembrolizumab and nivolumab; PDL1 inhibitors, atezolizumab (formerly MPDL3280A), durvalumab (MEDI4736), avelumab and the monoclonal antibody PDR001; 4-1BB ligand inhibitors, urelumab and utomilumab PF05082566; OX40 agonist, monoclonal antibody MEDI6469; glucocorticoid-inducible tumor necrosis factor receptor (GITR) inhibitor monoclonal antibody TRX518; CD27 inhibitor, varlilumab; TNFRSF25-TL1A inhibitors; CD40 agonist, monoclonal antibody CP 870893; inhibitors HVEM-LIGHTLTA and HVEM-BTLA-CD160; LAG3 inhibitors, monoclonal antibody BMS 986016; TIM3 inhibitors; Siglec inhibitors; ICOS ligand agonists; B7-H3 inhibitor, enoblituzumab MGA271; B7-H4 inhibitors; VISTA inhibitors; HHLA2-TMIGD2 inhibitors; butyrophilin inhibitors; BTNL2 inhibitors; CD244---CD48 inhibitors; inhibitors of members of the TIGIT and PVR families; KIR inhibitor, lirilumab; ILT and LIR inhibitors; NKG2D and NKG2A inhibitor, monalizumab IPH2201; MICA and MICB inhibitors; CD244 inhibitors; CSF1R inhibitors, emactuzumab, cabiralizumab, pexidartinib, ARRY382 HBLZ945; IDO inhibitor, (3E)-3-[(3-bromo-4-fluoroanilino)nitrosomethylidene]-4-[2-(sulfamoylamino)ethylamino]-1,2,5-oxadiazole INCB024360; TGFe inhibitor, galunisertib; adenosine-CD39-CD73 inhibitors; CXCR4-CXCL12 inhibitors, ulocuplumab and (3S,6S,9S, 12R, 17R,20S,23S,26S,29S,34aS)-N-((S)-1-amino-5-guanidino-1-oxopentane-2- yl)-26,29-bis(4aminobutyl)-17-((S)-2-((S)-2-((S)-2-(4-fluorobenzamido)-5-guanidinopentanamido)-5-guanidinopentanamido) -3 -(naphthalen-2-yl)propanamido)-6-(3-guanidinopropyl)-3,20-bis(4-hydroxybenzyl)- 1,4,7,10,18,21,24,27,30-nonaoxo-9,23-bis(3-ureidopropyl)triacontahydro-1H,16H-pyrrolo[2,1 p] [1,2]dithia[ 5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-12-carboxamide BKT140; phosphatidylserine inhibitors, bavituximab; SIRPA-CD47 inhibitor, monoclonal antibody CC 90002; VEGF inhibitor, bevacizumab; and or the neuropilin inhibitor, monoclonal antibody MNRP1685A. 13. The method according to claim 11, wherein the therapeutic agent is temozolomide. 14. The method according to claim 1, additionally providing for the use of radioactive irradiation. 15. The method according to claim 1, further comprising the administration of temozolomide and the use of radioactive irradiation. 16. The method of claim 10, wherein the additional therapeutic agent is selected from an AKT inhibitor, an alkylating agent, polytransretinoic acid, an antiandrogen, azacitidine, a BCL2 inhibitor, a BCL-XL inhibitor, a BCR-ABL inhibitor, a BTK inhibitor, a BTK/LCK/LYN inhibitor, CDK1/2/4/6/7/9 inhibitor, CDK4/6 inhibitor, CDK9 inhibitor, CBP/p300 inhibitor, EGFR inhibitor, endothelin receptor antagonist, ERK inhibitor, farnesyltransferase inhibitor, FLT3 inhibitor, glucocorticoid receptor agonist, FIDM2 inhibitor, inhibitor histone deacetylase, IKKe inhibitor, immunomodulatory drug (IMiD), ingenol, radiation exposure, ITK inhibitor, JAK1/JAK2/JAK3/TYK2 inhibitor, MEK inhibitor, midostaurin, MTOR inhibitor, PI3 kinase inhibitor, dual PI3/MTOR kinase inhibitor, proteasome inhibitor , protein kinase C agonist, SUV39H1 inhibitor, TRAIL, VEGFR2 inhibitor, Wnt/β-catenin signaling pathway inhibitor, decitabine and anti-CD20 monoclonal antibody. 17. Method for treating glioblastoma in a patient in need comprising administering to a patient 150 mg once daily or twice daily of a compound represented by the formula, or a pharmaceutically acceptable salt thereof. 18. The method of claim 17, further comprising administering an anticancer therapeutic agent, an anticancer biological agent, an immune checkpoint inhibitor, or a chemotherapeutic agent. 19. The method according to claim 18, where the therapeutic agent is selected from a cytotoxic agent, cispla - 18 045102 tina, doxorubicin, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, lonafarnib, tipifarnib, 4-((5-((4-(3-chlorophenyl) -3-oxopiperazin-1 -yl)methyl)-1 H-imidazol-1 -yl)methyl)benzonitrile hydrochloride, (R)-1 -((1 H-imidazol-5 -yl)methyl)-3-benzyl- 4-(thiophen-2-ylsulfonyl)-2,3,4,5-tetrahydro-1 H-benzodiazepine-7-carbonitrile, cetuximab, imatinib, interferon alpha-2b, pegylated interferon alpha-2b, combinations with aromatase, gemcitabine, uramustine, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylene thiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, leucovorin, oxaliplatin, pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17a-ethinyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone , megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, 17αhydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide acetate, flutamide, toremifene citrate, goserelin acetate, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, vinorel bina, anastrozole, letrozole, capecitabine, raloxifene, droloxafine, hexamethylmelamine, bevacizumab, trastuzumab, tositumomab, bortezomib, ibritumomab tiuxetan, arsenic trioxide, porfimer sodium, cetuximab, thiotepa, altretamine, fulvestrant, exemestane, rituximab, alemtuzumab, dexamethasone, bicalutamide, chlorambucil or valrubicin. 20. The method of claim 18, wherein the immune checkpoint inhibitor is selected from CTLA4 inhibitors, ipilimumab and tremelimumab; PD1 inhibitors, pembrolizumab and nivolumab; PDL1 inhibitors, atezolizumab (formerly MPDL3280A), durvalumab (formerly MEDI4736), avelumab and the monoclonal antibody PDR001; 4-1BB ligand inhibitors, urelumab and utomilumab PF 05082566; OX40 ligand agonist, monoclonal antibody MEDI6469; glucocorticoid-inducible tumor necrosis factor receptor (GITR) inhibitor monoclonal antibody TRX518; CD27 inhibitor, varlilumab; TNFRSF25-TL1A inhibitors; CD40 ligand agonist, monoclonal antibody CP 870893; inhibitors HVEM-LIGHT-LTA and HVEM-BTLA-CD160; LAG3 inhibitors, monoclonal antibody BMS 986016; TIM3 inhibitors; Siglec inhibitors; ICOS ligand agonists; B7-H3 inhibitor, enoblituzumab MGA271; B7-H4 inhibitors; VISTA inhibitors; HHLA2-TMIGD2 inhibitors; butyrophilin inhibitors; BTNL2 inhibitors; CD244---CD48 inhibitors; inhibitors of members of the TIGIT and PVR families; KIR inhibitor lirilumab; ILT and LIR inhibitors; NKG2D and NKG2A inhibitor, monalizumab IPH2201; MICA and MICB inhibitors; CD244 inhibitors; CSF1R inhibitors, emactuzumab, cabiralizumab, pexidartinib, ARRY382 and BLZ945; IDO inhibitor, (3E)-3-[(3-bromo4-fluoroanilino)-nitrosomethylidene]-4-[2-(sulfamoylamino)ethylamino]-1,2,5-oxadiazole INCB024360; TGFe inhibitor galunisertib; adenosine-CD39-CD73 inhibitors; CXCR4-CXCL12 inhibitors ulocuplumab and (3S,6S,9S,12R,17R,20S,23S,26S,29S,34aS)-N-((S)-1-amino-5-guanidino-1-oxopentan-2yl)- 26,29-bis(4-aminobutyl)-17-((S)-2-((S)-2-((S)-2-(4-fluorobenzamido)-5-guanidinopentanamido)-5guanidinopentanamido)-3- (naphthalen-2-yl)propanamido)-6-(3-guanidinopropyl)-3,20-bis(4-hydroxybenzyl)-1,4,7,10,18,21,24,27,30-nonaoxo-9 ,23-bis(3-ureidopropyl)triacontahydro-1H,16H-pyrrolo[2,1p][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-12- carboxamide BKT140; phosphatidylserine inhibitors, bavituximab; SIRPA-CD47 inhibitor, monoclonal antibody CC 90002; VEGF inhibitors, bevacizumab; and or the neuropilin inhibitor, monoclonal antibody MNRP1685A. 21. The method according to claim 19, wherein the therapeutic agent is temozolomide. 22. The method according to claim 17, additionally providing for the use of radioactive irradiation. 23. The method according to claim 17, further comprising the administration of temozolomide and the use of radioactive irradiation. 24. The method of claim 18, wherein the additional therapeutic agent is selected from an AKT inhibitor, an alkylating agent, a polytransretinoic acid, an antiandrogen, azacitidine, a BCL2 inhibitor, a BCL-XL inhibitor, a BCR-ABL inhibitor, a BTK inhibitor, a BTK/LCK/LYN inhibitor, CDK1/2/4/6/7/9 inhibitor, CDK4/6 inhibitor, CDK9 inhibitor, CBP/p300 inhibitor, EGFR inhibitor, endothelin receptor antagonist, ERK inhibitor, farnesyltransferase inhibitor, FLT3 inhibitor, glucocorticoid receptor agonist, HDM2 inhibitor, inhibitor histone deacetylase, IKKe inhibitor, immunomodulatory drug (IMiD), ingenol, radiation exposure, ITK inhibitor, JAK1/JAK2/JAK3/TYK2 inhibitor, MEK inhibitor, midostaurin, MTOR inhibitor, PI3 kinase inhibitor, dual PI3/MTOR kinase inhibitor, proteasome inhibitor , protein kinase C agonist, SUV39H1 inhibitor, TRAIL, VEGFR2 inhibitor, Wnt/β-catenin signaling pathway inhibitor, decitabine and anti-CD20 monoclonal antibody. 25. A method of treating PDGFRa-mediated gastrointestinal stromal tumors in a patient in need thereof, comprising administering to the patient 150 mg once daily or - 19 045102 twice daily compound represented by the formula or a pharmaceutically acceptable salt thereof. 26. The method of claim 25, further comprising administering an anticancer therapeutic agent, an anticancer biological agent, an immune checkpoint inhibitor, or a chemotherapeutic agent. 27. The method according to claim 26, where the therapeutic agent is selected from a cytotoxic agent, cisplatin, doxorubicin, etoposide, irinotecan, topotecan, paclitaxel, docetaxel, epothilones, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, lonafarnib, tipifarnib, 4- ((5 -((4-(3-chlorophenyl)-3-oxopiperazin-1 -yl)methyl)-1 H-imidazol-1 -yl)methyl)benzonitrile hydrochloride, (R)-1-((1H-imidazole -5-yl)methyl)-3-benzyl-4-(thiophen-2-ylsulfonyl)-2,3,4,5-tetrahydro-1H-benzodiazepine-7-carbonitrile, cetuximab, imatinib, interferon alpha-2b, pegylated interferon alpha-2b, combinations with aromatase, gemcitabine, uramustine, chlormethine, ifosfamide, melphalan, chlorambucil, pipobromane, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cytarabine, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate , leucovorin, oxaliplatin, pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17a-ethinylestradiol, diethylstilbestrol, dough sterone, prednisone, fluoxymesterone, dromostanolone propionate , testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, 17αhydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide acetate, flutamide, toremifene citrate, goserelin acetate, carboplatin, hydroxymo chevin, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, vinorelbine, anastrozole, letrozole , capecitabine, raloxifene, droloxafine, hexamethylmelamine, bevacizumab, trastuzumab, tositumomab, bortezomib, ibritumomab tiuxetane, arsenic trioxide, sodium porfimer, cetuximab, thiotepa, altretamine, fulvestrant, exemestane, rituximab, alemtuzumab , dexamethasone, bicalutamide, chlorambucil or valrubicin. 28. The method of claim 26, wherein the immune checkpoint inhibitor is selected from CTLA4 inhibitors, ipilimumab and tremelimumab; PD1 inhibitors, pembrolizumab and nivolumab; PDL1 inhibitors, atezolizumab (formerly MPDL3280A), durvalumab MEDI4736, avelumab and the monoclonal antibody PDR001; 4-1BB ligand inhibitors, urelumab and utomilumab PF05082566; OX40 ligand agonist, monoclonal antibody MEDI6469; glucocorticoid-inducible tumor necrosis factor receptor (GITR) inhibitor monoclonal antibody TRX518; CD27 inhibitor, varlilumab; TNFRSF25-TL1A inhibitors; CD40 ligand agonist, monoclonal antibody CP870893; inhibitors HVEM-LIGHT-LTA and HVEM-BTLA-CD160; LAG3 inhibitors, monoclonal antibody BMS 986016; TIM3 inhibitors; Siglec inhibitors; ICOS ligand agonists; B7-H3 inhibitor, enoblituzumab MGA271; B7-H4 inhibitors; VISTA inhibitors; HHLA2-TMIGD2 inhibitors; butyrophilin inhibitors; BTNL2 inhibitors; CD244---CD48 inhibitors; inhibitors of members of the TIGIT and PVR families; KIR inhibitor, lirilumab; ILT and LIR inhibitors; NKG2D and NKG2A inhibitor, monalizumab IPH2201; MICA and MICB inhibitors; CD244 inhibitors; CSF1R inhibitor, emactuzumab, cabiralizumab, pexidartinib, AMG382 and BLZ945; IDO inhibitor, (3E)-3-[(3-bromo-4fluoroanilino)nitrosomethylidene]-4-[2-(sulfamoylamino)ethylamino]-1,2,5-oxadiazole INCB024360; TGFe inhibitor galunisertib; adenosine-CD39-CD73 inhibitors; CXCR4-CXCL12 inhibitors ulocuplumab and (3S,6S,9S,12R,17R,20S,23S,26S,29S,34aS)-N-((S)-1-amino-5-guanidino-1-oxopentan-2yl)- 26,29-bis(4-aminobutyl)-17-((S)-2-((S)-2-((S)-2-(4-fluorobenzamido)-5-guanidinopentanamido)-5guanidinopentanamido)-3- (naphthalen-2-yl)propanamido)-6-(3-guanidinopropyl)-3,20-bis(4-hydroxybenzyl)-1,4,7,10,18,21,24,27,30-nonaoxo-9 ,23-bis(3-ureidopropyl)triacontahydro-1H,16H-pyrrolo[2,1p][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-12- carboxamide BKT140; phosphatidylserine inhibitors, bavituximab; SIRPA-CD47 inhibitor, monoclonal antibody CC 90002; VEGF inhibitor, bevacizumab; and or the neuropilin inhibitor, monoclonal antibody MNRP1685A. 29. The method according to claim 27, wherein the therapeutic agent is temozolomide. 30. The method according to claim 25, further comprising the use of radioactive irradiation. 31. The method according to claim 25, further comprising the administration of temozolomide and the use of radioactive irradiation. 32. The method of claim 26, wherein the additional therapeutic agent is selected from an AKT inhibitor, an alkylating agent, a polytransretinoic acid, an antiandrogen, azacitidine, a BCL2 inhibitor, a BCL-XL inhibitor, a BCR-ABL inhibitor, a BTK inhibitor, a BTK/LCK/LYN inhibitor, CDK1/2/4/6/7/9 inhibitor, CDK4/6 inhibitor, CDK9 inhibitor, CBP/p300 inhibitor, EGFR inhibitor, anti- - 20 045102 endothelin receptor gonist, ERK inhibitor, farnesyltransferase inhibitor, FLT3 inhibitor, glucocorticoid receptor agonist, HDM2 inhibitor, histone deacetylase inhibitor, ΙΚΚβ inhibitor, immunomodulatory drug (IMiD), ingenol, radiation exposure, ITK inhibitor, JAK1/JAK inhibitor 2/JAK3 /TYK2, MEK inhibitor, midostaurin, MTOR inhibitor, PI3 kinase inhibitor, dual PI3/MTOR kinase inhibitor, proteasome inhibitor, protein kinase C agonist, SUV39H1 inhibitor, TRAIL, VEGFR2 inhibitor, Wnt/β-catenin signaling pathway inhibitor, decitabine and monoclonal antibodies to CD20. 33. The method according to claim 1, including administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl ] -2-fluorophenyl]-3-phenylurea once a day. 34. Method according to and. 17, comprising administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl]-2-fluorophenyl] -3-phenylurea once a day. 35. The method according to claim 25, including administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl ]-2-fluorophenyl]-3-phenylurea once a day. 36. The method according to claim 1, including administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl ]-2-fluorophenyl]-3-phenylurea twice a day. 37. The method according to claim 17, including administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl ]-2-fluorophenyl]-3-phenylurea twice a day. 38. The method according to claim 25, including administering to the patient 150 mg of 1-[4-bromo-5-[1-ethyl-7-(methylamino)-2oxo-1,2-dihydro-1,6-naphthyridin-3-yl ]-2-fluorophenyl]-3-phenylurea twice a day.