Triazine derivatives as reversible and irreversible covalent inhibitors of PI3K

Abstract

The present invention relates to novel triazine compounds that contain chemically reactive groups (warheads) and behave as reversible and irreversible covalent inhibitors. Linkers are introduced to target solvent-exposed cysteines more than 10 Å distal from the core reversible inhibitor. Different exit vectors are explored to modulate the inhibitor's intrinsic reactivity and the efficiency of covalent bond formation. We disclose novel optimized covalent modifiers of phosphoinositide 3-kinase alpha (PI3Kα), an enzyme frequently altered in human malignancies. The compounds of the present invention can be utilized as therapeutic agents and chemical probes useful for investigating the role of PI3K isoforms in cancer and metabolism, and for treating PI3Kα-driven cancers and malformations.

Description

[Technical Field]

[0001] This invention relates to novel triazine compounds containing chemically reactive groups (warheads) that serve as therapeutic agents and chemical probes useful for regulating cellular activities such as signal transduction, proliferation, differentiation, and cell death. The compounds of this invention regulate kinase activity, particularly the activity of phosphoinositide 3-kinase (PI3K). [Background technology]

[0002] Protein kinases are involved in signaling events and regulate cell activation, growth, differentiation, survival, and migration in response to extracellular mediators or stimuli (including growth factors, cytokines, or chemokines).

[0003] Increased protein kinase activity is associated with many diseases, including cancer, inflammatory disorders, and metabolic and immune disorders. These can be caused directly or indirectly by dysfunction of regulatory mechanisms due to mutations, overexpression, or improper control of enzyme activity.

[0004] The phosphoinositide 3-kinase (PI3K) signaling pathway plays a crucial role in many cellular processes, including cell growth, proliferation, and survival. The PI3K family is divided into three classes according to their amino acid sequence, homology, and substrate specificity. Class I PI3Ks are activated downstream of cell surface receptors, including receptor protein tyrosine kinases (RTKs), G protein-coupled receptors (GPCRs), and immunoglobulin receptors. Class IA PI3Ks are obligate heterodimers consisting of a catalytic subunit (p110α, p110β, or p110δ) and associated regulatory subunits (p85α, p85β, p50α, p55α, or p55γ). Class IB PI3Kγ acts downstream of GPCRs and consists of a catalytic subunit (p110γ) and an adapter subunit (p84 or p101). The cell surface receptor activates PI3K, producing PtdIn(3,4,5)P3, which acts as a docking site for protein kinase B (PKB / Akt) and 3-phosphoinositide-dependent protein kinase 1 (PDK1). This leads to phosphorylation of the kinase domain at two regulatory sites: Thr308 by PDK1 and Ser473 by mTOR complex 2 (mTORC2). Overactivation of the PI3K / mTOR pathway can occur at multiple levels of this signaling cascade, ultimately promoting cancer growth and progression. Loss or inactivation of tumor suppressor phosphatases and tensin homologs (PTENs), mutations or amplification of cell surface receptors, and the presence of activating hotspot mutations in PIK3CA play important roles in human carcinogenesis. Furthermore, overactivation of the PI3K / mTOR axis is associated with resistance to multiple cancer treatments. Therefore, PI3K inhibitors are considered to have high value in cancer treatment.

[0005] Considerable efforts have been devoted to the development of drugs targeting PI3K signaling, and many of them are currently being evaluated in clinical trials. Selective PI3Kα inhibitors may be beneficial for PIK3CA mutant tumors and PIK3CA-related overgrowth syndrome (PROS), and can minimize the on-target metabolic side effects of pan-PI3K inhibitors. BYL719 / Alpelisib / PIKRAY from Novartis and GDC-0032 / Taselisib from Genentech act as reversible modifiers and are claimed as PI3Kα-selective inhibitors. However, at the concentrations required for in vivo experiments, they cannot distinguish PI3K isoforms. Only one PI3Kα covalent inhibitor called CNX-1351 is currently available, but it exhibits limited in vitro and cellular potency, low water solubility, and metabolic instability.

[0006] This patent application describes certain triazine derivatives having PI3K inhibitory activity and acting as irreversible and reversible covalent modifiers, as well as their use as pharmaceuticals. The compounds covered herein have significant advantages with respect to potency, metabolic stability, and drug-like properties compared to CNX-1351. Furthermore, this patent application describes linkers for the development of covalent kinase inhibitors.

Summary of the Invention

[0007] The present invention relates to novel triazine-based compounds acting as irreversible modifiers, as well as their use as therapeutic agents and chemical probes.

[0008] The first aspect of the present invention is a compound of formula (IV), particularly formula (IVa)

Chemical Formula

[0009] A second aspect of the present invention relates to a compound according to the first aspect of the present invention for use in the treatment of diseases.

[0010] A third aspect of the present invention relates to compounds according to the first aspect of the present invention for use in the treatment of tumor diseases, overgrowth syndromes, neurological disorders, and immune disorders.

[0011] A fourth aspect of the present invention is the intermediate of formula (VI) [ka] (In the formula, R1, R2, R4, L2, W2, U, n and W1 are defined as above, Z is -OH, Br, COOH, -C(OH)NH2. Regarding.

[0012] Detailed description of the invention Herein, certain embodiments of the present invention are described in detail, and examples thereof are illustrated in the attached structures and formulas. The present invention is described in conjunction with the listed embodiments, but it will be understood that they are not intended to limit the present invention to those embodiments. Rather, the present invention is intended to cover all substitutes, modifications, and equivalents that may fall within the scope of the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is by no means limited to the methods and materials described herein.

[0013] definition As used herein, the term "alkyl" refers to a saturated linear monovalent hydrocarbon group having 1 to 5 carbon atoms (C1 to C5). Examples of alkyl groups include, but are not limited to, methyl, ethyl, 1-propyl (n-propyl), and 1-butyl (n-butyl).

[0014] The terms “heterocyclic,” “heterocyclyl,” and “heterocyclic ring” are used interchangeably herein and refer to a saturated or unsaturated carbocyclic radical of 4 to 6 ring atoms, in which at least one ring atom is a heteroatom, in particular nitrogen, the remaining ring atoms are carbon atoms, and one or more ring atoms are optionally independently substituted with one or more substituents selected in particular from -CH3 and -F.

[0015] The term "chiral" refers to a molecule that cannot be superimposed on its mirror image partner, while the term "achiral" refers to a molecule that can be superimposed on its mirror image partner.

[0016] The term "stereoisomer" refers to compounds that have the same chemical structure but differ in the arrangement of atoms or groups in space.

[0017] A "diastereomer" refers to a stereoisomer of a compound that has two or more chirality centers that are not mirror images of each other. Diastereomers have different physical properties, such as melting point, boiling point, spectral properties, and chemical and biological reactivity. Mixtures of diastereomers can be separated under high-resolution analytical procedures such as electrophoresis and chromatography.

[0018] An "enantiomer" refers to two stereoisomers of a compound that are mirror images of each other and cannot be superimposed.

[0019] The definitions and conventions of stereochemistry used herein generally follow the McRaw-Hiff Dictionary of Chemical Terms (1984) edited by SP. Parker, McGraw-Hill Book Company, New York; and "Stereochemistry of Organic Compounds" by Eliel, E. and Wilen, S., John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain a chiral or asymmetric center and therefore may exist in various stereoisomeric forms. All stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers, and atropisomers, and mixtures thereof, such as racemic mixtures, are intended to form part of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of polarization. When describing optically active compounds, the prefixes R and S are used to indicate the absolute configuration of the molecule around its chiral center. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Certain stereoisomers are sometimes called enantiomers, and mixtures of such isomers are often called enantiomer mixtures. A 50:50 mixture of enantiomers is called a racemic mixture or racemate. The term "tautomer" or "tautomeristic form" refers to structural isomers of different energies that can be interconverted across a low-energy barrier. For example, proton tautomers include interconversions by proton transfer, such as keto-enol isomerization and imine-enamine isomerization.

[0020] The term "enone" refers to an α,β-unsaturated carbonyl, a type of organic compound consisting of an alkene conjugated to a ketone. The simplest enones are methyl vinyl ketone (butenone) or CH2=CHCOCH3. These are electrophilic at both the carbonyl and β-carbons. Depending on the conditions, either site is attacked by a nucleophile. Addition to an alkene is called Michael addition and is used in this invention to covalently modify cysteine ​​862 in PI3Kα.

[0021] The term "acrylamide" is derived from acrylic acid and refers to an amide having the general chemical formula CH2=CHC(O)NH2. Acrylamide is used in the compounds of the present invention and undergoes Michael addition with cysteine ​​862 in PI3Kα.

[0022] The term "PI3K" refers to phosphoinositide 3-kinase.

[0023] The terms "PI3K alpha," "PI3Kα," or "p110a protein" refer to the subunit of PI3K encoded by the PI3KCA gene.

[0024] The terms “irreversible” or “irreversible inhibitor” refer to inhibitors that can covalently bind to PI3 kinase substantially irreversibly, whereas reversible inhibitors can bind to the kinase (but generally cannot form a covalent bond) and are therefore able to dissociate from PI3 kinase. Once covalent bond formation occurs, an irreversible inhibitor remains substantially bound to the kinase. Methods for determining whether a compound is acting as an irreversible inhibitor are known to those skilled in the art. Such methods include, but are not limited to, enzymatic kinetic analysis of the inhibitory profile of a compound by the kinase, the use of mass spectrometry of a protein-drug target modified in the presence of an inhibitor compound, the use of X-ray crystallography to analyze the complex between the protein-drug target and the inhibitor compound, intermittent exposure also known as a “washout” experiment, and other methods known to those skilled in the art.

[0025] The term "reversible covalent bonding" refers to inhibitors that covalently modify target cysteine, but the free energy difference separating non-covalent reactants from the bonded product is close to equilibrium, the activation barrier is relatively low, and as a result, the reverse reaction that cleaves the chemical bond occurs easily (examples include nitrile-based reversible covalent inhibitors).

[0026] The terms "warhead" or "warhead group" refer to a functional group present in the compound of the present invention that covalently binds to an amino acid residue (e.g., cysteine, lysine, histidine, or other residues that can be covalently modified) present in the binding pocket of a target protein, thereby irreversibly inhibiting the protein. The warhead group is essential for covalent and irreversible inhibition of the protein.

[0027] The term "inhibitor" is defined as a compound that binds to and inhibits PI3 kinase with measurable affinity. In certain embodiments, the inhibitor is IC 50 and / or rate constant (k) for irreversible deactivation inact Characterized by:

[0028] The term "CNX-1351" refers to 1-[4-[[2-(1H-indazole-4-yl)-4-(4-morpholinyl)thieno[3,2-d]pyrimidine-6-yl]methyl]-1-piperazinyl]-6-methyl-5-hepten-1,4-dione (CAS 1276105-89-5).

[0029] When used herein, the expression "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of the compound of the present invention. If the compound of the present invention is a base, a desired pharmaceutically acceptable salt can be prepared by any suitable method available in the art, for example, by treating a free base with an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, methanesulfonic acid, or phosphoric acid, or with an organic acid such as pyranosidylic acid such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, glucuronic acid, or galacturonic acid, alpha hydroxy acids such as citric acid or tartaric acid, amino acids such as aspartic acid or glutamic acid, aromatic acids such as benzoic acid or cinnamic acid, or sulfonic acids such as p-toluenesulfonic acid or ethanesulfonic acid.

[0030] The term "protecting group" refers to a substituent commonly used to block or protect a specific functional group during the reaction of other functional groups on a compound. For example, an "amino protecting group" is a substituent attached to an amino group that blocks or protects an amino functional group in a compound. Suitable amino protecting groups include acetyl, trifluoroacetyl, tert-butoxycarbonyl (BOC), benzyloxycarbonyl, and 9-fluorenylmethyleneoxycarbonyl (Fmoc). For a general explanation of protecting groups and their uses, see Protective Groups in Organic Synthesis by TW Greene, John Wiley & Sons, New York, 1991.

[0031] The terms “compounds of the present invention” and “compounds of formula (I, II, III)” include their stereoisomers, geometric isomers, tautomers, solvates, pharmaceutically acceptable salts, and solvates of those salts.

[0032] Detailed explanation The present invention relates to novel triazine-based compounds that act as reversible or irreversible modifiers of PI3 kinase, particularly PI3Kα, and their use as therapeutic agents and chemical probes.

[0033] The compounds of the present invention have higher water solubility (>30-fold), higher potency in vitro and intracellularly (>7-fold), and higher metabolic stability compared to the known inhibitor CNX-1351.

[0034] An important aspect of the present invention relates to the favorable reaction parameters in preferred molecules (particularly inhibitors characterized by a warhead that forms an acrylamide moiety) that result in highly selective target binding and reduced or negligible off-target reactivity, i) The on-target reaction of a covalent inhibitor is described by the inhibitor dissociation constant K that accounts for the reversible equilibrium of complex formation between the enzyme (E) and the inhibitor (I) i E + I ←→ E~I and the reaction rate k that defines the formation of a covalent inhibitor-enzyme complex (EI) between the inhibitor and the enzyme inact as defined by E~I → EI ii) The off-target reaction with ubiquitous sulfhydryls (S) and other nucleophiles present in physiological environments such as cells and body fluids is driven by the intrinsic reactivity of the warhead defined by k chem and results in the formation of sulfhydryl adducts (SI) by driving the conversion of S and I S + I → SI

[0035] A high intrinsic warhead reactivity (high k chem value) leads to the formation of undesirable sulfhydryl adducts that result in compound loss, covalent modification of cellular components, toxicity, and anti-compound immune responses:

[0036] As exemplified in Table 1, preferred molecules have low k chem , low K i , high kinact , high k inact / K i Shows the ratio.

[0037] A first aspect of the present invention relates to compounds of formula (IV), particularly formula (IVa). [ka] [In the formula, X is CH or N, especially N. Y is H or F, especially H. R1 and R2 are independently H, CH3, cyclopropyl, -F, -CH2-F, -CH2-CH2-F, -CN, [ka] (In the formula, R5 is F or CH3, and R6 is C 1~6 It is an alkyl group, and z is selected from 0, 1, or 2. · ·R3 is C 1~3 Either alkyl or two residues R3 are cross-linked -(CH2) r - forms, where r is 1, 2, or 3, in particular r is 1 or 2, v is 0, 1, 2, 3, or 4. R4 is H, F, or -CN. L2 is [ka] (In the formula, R5 is C 1~3 The moiety is selected from alkyl, F, -CH2CN or -CN, where t is 0, 1 or 2. W1 is either CO or CH2. W2 is selected from O, CH2, and CO. U is selected from O, CH2, CO, NH and N(CH3), especially O, CH2, NH and N(CH3), n is either 1 or 2. or relating to its prodrugs, metabolites, tautomers, solvates, or pharmaceutically acceptable salts, in particular to its tautomers, solvates, or pharmaceutically acceptable salts.

[0038] The inhibitor according to the present invention comprises a polycyclic scaffold and a so-called warhead connected to the scaffold via a linker. The scaffold comprises three heterocycles, namely morpholinyl, piperazinyl, and a triazine moiety substituted with a pyridinyl or pyrimidinyl moiety. The pyridinyl or pyrimidinyl moiety is substituted with a fluorinated methyl and amine moiety. The morpholinyl moiety is optionally substituted. Piperazinyl is L2, W2, U, C 1~2 It is bonded to a linker composed of alkyl and W1.

[0039] The molecule presents a specific reactive functional group known as a “warhead.” As used herein, the terms “warhead” or “warhead group” refer to a functional group present in the compound of the present invention that covalently binds to an amino acid residue (e.g., cysteine, lysine, histidine, or other residues that can be covalently modified) present in the binding pocket of a target protein, thereby irreversibly inhibiting the protein.

[0040] The inhibitor according to the present invention may be covalently bonded to Cys862 of PI3Kα. Such inhibitors are characterized by a carbon-carbon double bond within the warhead. The stability of the bond can be modulated by substituents R1, R2, and R4 to achieve inhibitors that form a stable covalent bond or a reversible covalent bond.

[0041] The compounds disclosed herein exhibit good stability, along with good PI3 kinase inhibition, particularly good PI3Kα inhibition.

[0042] In certain embodiments, R1 is H, CH3, or -CH2F.

[0043] In certain embodiments, R2 is H or cyclopropyl.

[0044] In certain embodiments, R2 is cyclopropyl and R4 is -CN.

[0045] In a particular embodiment, R3 is C 1~3 Alkyl groups, especially CH3.

[0046] In certain embodiments, v is 0, 1, or 2, more specifically, 0 or 1.

[0047] In certain embodiments, the compound is a compound of formula (V), particularly (Va). [ka] (In the formula, X, Y, R1, R2, R3, R4, W1, n, U, W2, L2 are defined as above. (v is either 0 or 1) That is the case.

[0048] In a particular embodiment, the compound according to the first aspect of the present invention is a compound of formula (I) or (II). [ka] [In the formula, R is either H or CH3. R1 is either H or CH3. L2 is [ka] (In the formula, R5 is C 1~3 [The part is selected from alkyl, F, -CH2CN or -CN, where t is 0, 1 or 2] [ka] [In the formula, R3 is either H or CH3. L2 is [ka] (In the formula, R5 is C 1~3 The moiety is selected from alkyl, F, -CH2CN or -CN, where t is 0, 1 or 2. If U is CH2, then W2 is O, or if U is O, then W2 is CH2; If W2 is CH2, then U is O, or if W2 is O, then U is CH2; R1 is either H or -CH2-F. R2 is either H or cyclopropyl. R4 is either H or F. These include the reversible analogues, prodrugs, metabolites, tautomers, solvates, and pharmaceutically acceptable salts (I, II) thereof.

[0049] In a particular embodiment, the compound according to the first aspect of the present invention is a compound of formula (Ia). [ka] [In the formula, R is either H or CH3; W1 consists of CO and CH2; L2 is [ka] (In the formula, R5 is C 1~3 [The part is selected from alkyl, F, -CH2CN or -CN, where t is 0, 1 or 2] That is the case.

[0050] Furthermore, the present invention relates to the synthesis of compounds of formula (I, II) as defined above, including tautomers, solvates, intermediates, prodrugs, and salts of the aforementioned compounds.

[0051] A second aspect of the present invention relates to a compound according to the first aspect of the present invention for use in the treatment of diseases.

[0052] Another aspect of the present invention relates to compounds according to the first aspect of the present invention for use in the treatment of diseases caused by activating mutations in the PI3KCA gene or activation of class I PI3K, particularly PI3Kα. Activation of class I PI3K, particularly PI3Kα, can occur by activating mutations in cell surface receptors, upstream overexpression or mutated upstream activators, or PI3K interaction and regulatory proteins, including the PI3KCA gene or the gene products of PIK3R1, PIK3R1, and PIK3R1.

[0053] A third aspect of the present invention relates to compounds according to the first aspect of the present invention for use in the treatment of tumor diseases, overgrowth syndromes, neurological diseases, and immunological diseases.

[0054] In certain embodiments, the tumor is a solid tumor, and / or the tumor disease is selected from lymphoma and leukemia.

[0055] In certain embodiments, the compounds according to the first aspect of the present invention are used for proliferative diseases; benign or malignant tumors; tumors arising from sarcomas; lungs; bronchi; prostate; breast; pancreas; gastrointestinal cancer; colon; rectum; colon cancer; colorectal adenoma; thyroid; liver; intrahepatic bile ducts; hepatocytes; adrenal glands; stomach; gastric; glioma; glioblastoma; endometrium; melanoma; kidney; renal pelvis; bladder; uterine body; cervix; vagina; ovaries; multiple myeloma; esophagus; Treatment of leukemia; acute myeloid leukemia; chronic myeloid leukemia; lymphocytic leukemia; myeloid leukemia; brain; brain cancer; oral cavity and pharynx; larynx; small intestine; non-Hodgkin lymphoma; melanoma; choriocolonic adenoma; neoplasms; epithelial neoplasms; lymphoma; treatment of breast cancer; basal cell carcinoma; squamous cell carcinoma; actinic keratosis; tumor diseases including solid tumors; tumors of the neck or head; polycythemia vera; essential thrombocythemia; myelofibrosis with myelogenesis; and Waldenström macroglobulinemia; sporadic cancers including breast cancer, thyroid cancer, uterine cancer, and other cancers that appear in patients with these syndromes. It is intended for use in the treatment of ocular neovascularization and multiple hamartoma syndrome; fibrolipid hyperplasia (also called fibrolipid hypergrowth), CLOVES syndrome, cerebral capillary malformation syndrome (MCAP syndrome), hemiplegia-multiple lipomatosis syndrome (HHML syndrome), hemimegalencephaly, and PIK3CA-associated hypergrowth spectrum (PROS) related disorders, including facial and infiltrative lipomatosis in other organs; systemic vascular and lymphatic malformations, and tumor angiogenesis; ocular neovascularization and macular degeneration (AMD), proliferative and diabetic retinopathy (PDR), and retinopathy of prematurity (ROP).

[0056] The present invention also relates to such PI3K-targeted compounds and their pharmaceutical formulations as chemotherapeutic agents having anticancer activity that may be potentially useful in treating diseases, conditions, and / or disorders regulated by enhancement of cell division, growth, migration, adhesion, and metastasis. The compounds may inhibit tumor growth in mammals and may be useful in treating human cancer patients.

[0057] The present invention also refers to the ability to modulate the intrinsic reactivity of inhibitors by modifying the exit vector of the warhead (meta, ortho position of a 6, 5-membered ring, or 4-membered ring).

[0058] Furthermore, with respect to cancer treatment, the present invention also relates to the treatment of pathogenic cellular conditions caused by cellular hyperactivation using such compounds. Molecules targeting PI3K can be used to treat a variety of hyperproliferative diseases in order to treat or prevent diseases or conditions regulated by PI3K.

[0059] The present invention also relates to such PI3K target compounds as chemical probes for analyzing the role of PI3K isoforms in cancer and metabolism.

[0060] Furthermore, the present invention relates to methods or production processes for using such compounds for in vitro, in situ, and in vivo diagnostic procedures or treatments of mammalian cells, organisms, or related pathological conditions.

[0061] In further embodiments, the present invention relates to pharmaceutical compositions comprising compounds of formulas (I, II) as defined above, and methods for preventing or treating diseases or disorders modulated by PI3K, particularly for treating hyperproliferative disorders.

[0062] Further aspects of the present invention relate to the use of effective amounts of the compounds of formula (I, II) as defined above, either alone or in combination with standard treatments such as chemotherapy, radiotherapy, targeted therapy, or immunotherapy for diseases or disorders modulated by PI3K, particularly hyperproliferative disorders.

[0063] In a further embodiment, the present invention relates to a compound of formula (III) as a linker in the design and synthesis of covalent compounds targeting protein kinases. [ka] [In the formula, U is equivalent to NH, NCH3, O, or CH2; Y is equal to CO, O, or CH2; L2 is azetidine, pyrrolidine, or piperidine, and the arrow is in formula (III): [ka] [Shows the connection in] Regarding use.

[0064] Another aspect of the present invention includes methods for preparing, separating, and purifying the compounds defined above, including intermediates, prodrugs, and salts of the compounds of formula (I, II, III).

[0065] Another aspect of the present invention includes novel intermediates useful for preparing compounds of formulas (I, II, III) as defined above.

[0066] Another aspect of the present invention involves novel and improved properties of compounds of formula (I, II, III) with respect to CNX-1351. These properties include, but are not limited to, in vitro and cellular potency, metabolic stability, solubility, and drug-like properties.

[0067] Another aspect of the present invention involves modifying the intrinsic reactivity of an inhibitor by modifying the exit vector (ortho or meta of a 6-membered ring and a 5-membered ring, or a 4-membered ring exit vector).

[0068] The cysteine ​​residue of PI3Kα targeted by the irreversible inhibitor of the present invention is the non-conservative Cys862.

[0069] Medical treatments, dosage forms, and salts Similarly, a method for treating tumor diseases, overgrowth syndromes, neurological disorders and / or immune disorders in patients requiring such treatment, comprising the step of administering the compound described above to the patient, falls within the scope of the present invention.

[0070] Similarly, dosage forms are provided for the prevention or treatment of tumor diseases, overgrowth syndromes, neurological disorders and / or immune disorders, comprising compounds according to any of the above embodiments or models of the present invention.

[0071] Those skilled in the art will recognize that any specifically mentioned drug compound referred to herein may exist as a pharmaceutically acceptable salt of the drug. A pharmaceutically acceptable salt includes an ionized drug and an oppositely charged counterion. Non-limiting examples of pharmaceutically acceptable anionic salt forms include acetates, benzoates, besilates, bitatrates, bromides, carbonates, chlorides, citrates, edetates, edisylates, embonates, estrates, fumarates, gluceptates, glucons, hydrobroms, hydrochlorides, iodides, lactates, lactobionates, malates, maleates, mandelates, mesylates, methyl bromides, methyl sulfates, mucinates, napsylates, nitrates, pamosates, phosphates, diphosphates, salicylates, disalicylates, stearates, succinates, sulfates, tartrates, tosylates, triethiozides, and valersates. Non-limiting examples of pharmaceutically acceptable cationic salt forms include aluminum, benzathine, calcium, ethylenediamine, lysine, magnesium, meglumine, potassium, procaine, sodium, tromethamine, and zinc.

[0072] The dosage form may be enteral, e.g., nasal, buccal, rectal, transdermal, or oral, or in inhalation form or as a suppository. Alternatively, parenteral administration such as subcutaneous, intravenous, intrahepatic, or intramuscular injection may be used. Depending on the circumstances, pharmaceutically acceptable carriers and / or excipients may be present.

[0073] Topical administration is also within the scope of the advantageous use of the present invention. Those skilled in the art will know a wide range of possible recipes for providing topical formulations, as exemplified by Benson and Watkinson (eds.), Topical and Transdermal Drug Delivery: Principles and Practice (1st edition, Wiley 2011, ISBN-13: 978-0470450291); Guy and Handcraft, Transdermal Drug Delivery Systems: Revised and Expanded (2nd edition, CRC Press 2002, ISBN-13: 978-0824708610); and Osborne and Amann (eds.), Topical Drug Delivery Formulations (1st edition, CRC Press 1989; ISBN-13: 978-0824781835).

[0074] Pharmaceutical composition and administration Another aspect of the present invention relates to a pharmaceutical composition comprising the compound of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. In further embodiments, the composition comprises at least two pharmaceutically acceptable carriers, such as those described herein.

[0075] In certain embodiments of the present invention, the compounds of the present invention are typically formulated into pharmaceutical dosage forms to provide a drug with an easily controllable dosage and to give the patient a sophisticated and easily manageable product.

[0076] In embodiments of the present invention relating to the topical use of the compounds of the present invention, the pharmaceutical composition is formulated in a manner suitable for topical administration, such as an aqueous solution, suspension, ointment, cream, gel, or sprayable formulation for delivery by aerosol, etc., and comprises the active ingredient together with one or more solubilizers, stabilizers, isotonic enhancers, buffers, and preservatives known to those skilled in the art.

[0077] The pharmaceutical composition can be formulated for enteral administration, particularly for oral or rectal administration. Furthermore, the pharmaceutical composition of the present invention may be in solid form (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories) or liquid form (including, but not limited to, solutions, suspensions, or emulsions).

[0078] The pharmaceutical composition can be formulated for parenteral administration, for example, by IV injection, intradermal, subcutaneous, or intramuscular administration.

[0079] The administration regimen of the compounds of the present invention will vary depending on known factors, such as the pharmacodynamic characteristics of a particular drug and its mode and route of administration; the recipient's species, age, sex, health status, medical condition, and weight; the nature and severity of symptoms; the type of concomitant treatment; the frequency of treatment; the route of administration, the patient's renal and hepatic function, and the desired effect. In certain embodiments, the compounds of the present invention may be administered in once-daily doses, or the total daily dose may be administered in two, three, or four divided doses per day.

[0080] In certain embodiments, the pharmaceutical composition of the present invention may have a unit dose of approximately 1 to 1000 mg of the active ingredient for a subject weighing approximately 50 to 70 kg. The therapeutically effective dose of a compound, pharmaceutical composition, or combination thereof depends on the species, weight, age, and individual condition of the subject being treated, the disorder or disease, or its severity. A physician, clinician, or veterinarian with ordinary skill can easily determine the effective amount of each active ingredient necessary to prevent, treat, or inhibit the progression of the disorder or disease.

[0081] The pharmaceutical compositions of the present invention can be subjected to conventional pharmaceutical operations such as sterilization, and / or may contain conventional inert diluents, lubricants, or buffers, as well as adjuvants such as preservatives, stabilizers, wetting agents, emulsifiers, and buffers. They can be produced by standard processes, for example, by conventional mixing, granulation, dissolution, or freeze-drying processes. Many such procedures and methods for preparing pharmaceutical compositions are known in the art; see, for example, The Theory and Practice of Industrial Pharmacy, 4th edition, 2013 by L. Lachman et al. (ISBN 8123922892).

[0082] Manufacturing method and treatment method according to the present invention The present invention further includes, in a more recent embodiment, the use of compounds according to the first embodiment of the present invention as specified herein, or pharmaceutically acceptable salts thereof as specified in detail above, for use in a method of producing a pharmacopoeia for treating or preventing a condition selected from tumor diseases, overgrowth syndromes, neurological disorders and / or immune disorders.

[0083] Similarly, the present invention encompasses methods for treating patients diagnosed with diseases associated with tumor diseases, overgrowth syndromes, neurological disorders, and / or immune disorders. The method involves administering to the patient an effective amount of a compound specified herein (SPECIFY), or a pharmaceutically acceptable salt thereof as specified herein in detail.

[0084] For example, whenever a substitute for a single separable feature, such as an isotype protein or coding sequence, ligand type, or medical indication, is described herein as an “embodiment,” it should be understood that such substitutes may be freely combined to form a separate embodiment of the invention disclosed herein. Thus, any of the substitute embodiments for detectable labels may be combined with any of the substitute embodiments for ligands, and these combinations may be combined with any medical indicator or diagnostic method referred to herein.

[0085] The present invention is further illustrated by the following examples and drawings, from which further embodiments and advantages can be derived. These examples are illustrative and not intended to limit the scope of the present invention.

[0086] In a particular embodiment, a compound according to the first aspect of the present invention is selected from compounds 1 to 50 shown in the following table.

[0087] In a particular embodiment, a compound according to the first aspect of the present invention, selected from compounds 1 to 52 shown in the following table.

[0088] The most preferred compound is the following, represented by the formula: (The corresponding structure names were created using ChemBioDraw Ultra, version 16.0). [Table 1] [Table 2] [Table 3] [Table 4] [Table 5-1] [Table 5-2] [Table 5-3] [Table 5-4] [Table 5-5] [Table 6] [Table 7]

[0089] The most preferred linkers are the following compounds (and their corresponding enantiomers) shown by the formula: [Table 8] [Brief explanation of the drawing]

[0090] Summary data of several compounds of the present invention compared to CNX-1351: intrinsic reactivity and in vitro data (Table 1); cytotoxicity of some of the compounds of the present invention and CNX-1351 (Table 2). [Figure 1] Metabolic stability of rat liver microsomes 18, 25, and 33, as well as CNX-1351, using rat liver microsomes enhanced with the phase I metabolic cofactor NADPH.

[0091] example Preparation of the compound of the present invention The compounds of the present invention can be synthesized by synthetic routes including methods similar to those well known in the chemical art, particularly in view of the description contained herein. Starting materials are generally available from commercial sources or can be readily prepared using methods well known to those skilled in the art.

[0092] For illustrative purposes, Schemes 1-4 illustrate general methods for preparing the compounds and key intermediates of the present invention. For a more detailed description of the individual reaction steps, see the following examples in this specification. Those skilled in the art will recognize that other synthetic routes may be used to synthesize the compounds of the present invention. While specific starting materials and reagents are shown in the schemes and discussed below, other starting materials and reagents can be readily substituted to provide a variety of derivatives and / or reaction conditions. Furthermore, many of the compounds prepared by the methods described below can be further modified in view of this disclosure using conventional chemistry well known to those skilled in the art.

[0093] When preparing the compounds of the present invention, protection and deprotection of the remote functionality (e.g., primary or secondary amine) of the intermediate may be necessary. The need for such protection will depend on the nature of the remote functionality and the conditions of the preparation method. Suitable amino protecting groups include tert-butyloxycarbonyl (BOC) or N,N-dimethylformamidinyl. The need for such protection will be readily determined by those skilled in the art. For a general description of protecting groups and their use, see Protective Groups in Organic Synthesis by TW Greene, John Wiley & Sons, New York, 1991.

[0094] Scheme 1 [ka] Scheme 1 shows a general method for preparing inhibitor building blocks using nucleophilic aromatic substitution and Suzuki coupling. Reagents and conditions: (i) Et3N, DCM, -50°C, 3 hours; (ii) 1-boc-piperazine, DIPEA, EtOH, 0°C → room temperature, 5 hours; (iii) tert-butyl N-[5-bromo-4-(difluoromethyl)pyrimidine-2-yl]-N-[(tert-butoxy)carbonyl]carbamate, bis(pinacolate)diborone, AcOK, Pd(dppf)Cl2, dioxane, 95°C, 1.5 hours; (2) monochlorotriazine, XPhosPdG2 (catalyst), K3PO4, H2O, 100°C, o / n; (3) HCl, dioxane / H2O, 80°C, o / n.

[0095] Scheme 2 [ka] Scheme 2 shows a general method for preparing a covalent inhibitor of formula (I) having an amide spacer. R1 is H or CH3. Reagents and conditions: (i) building block or HCl salt, HCTU, DIPEA, DMF, 0°C → room temperature, 4-16 hours; (ii) HCl in dioxane (4M), THF, room temperature, 3-16 hours.

[0096] Scheme 3 [ka] Scheme 3 shows a method for preparing the compound of formula (II). First, an acid with an ether in the linker is synthesized by nucleophilic substitution, followed by hydrolysis of the ester using a strong base in water / THF. Reagents and conditions: (i) NaH (60% dispersion in mineral oil), DMF, 0°C, 15 min; (ii) LiOH (5 M in H2O), THF; (iii) Building block or HCl salt, HCTU, DIPEA, DMF, 0°C → room temperature, 2 hours; (iv) HCl (4 M) in dioxane, THF, room temperature, o / n.

[0097] Scheme 4 [ka] Scheme 4 shows a method for preparing the compound of formula (II). First, an acid with an ether in the linker is synthesized by nucleophilic substitution, followed by hydrolysis of the ester using a strong base in water / THF. Reagents and conditions: (i) NaH (60% dispersion in mineral oil), DMF, 0°C, 15 min; (ii) LiOH (5 M in H2O), THF; (iii) Building block or HCl salt, HCTU, DIPEA, DMF, 0°C → room temperature, 2 hours; (iv) HCl (4 M) in dioxane, THF, room temperature, o / n.

[0098] Separation method In the method for preparing the compounds of the present invention, it may be advantageous to separate the reaction products from each other and / or from the starting materials. The desired products from each step or series of steps are separated and / or purified to a desired degree of homogeneity by techniques common in the art. Typically, such separation involves extraction, crystallization from a solvent or solvent mixture, or chromatography. Chromatography may involve, for example, reversed-phase and normal-phase; high-pressure, medium-pressure and low-pressure liquid chromatography methods and apparatus; small-scale analysis; and any number of methods including preparative thin-layer or thick-layer chromatography, as well as techniques for small-scale thin-layer and flash chromatography.

[0099] The selection of an appropriate separation method depends on the properties of the materials involved, such as the presence or absence of polar functional groups in chromatography and the stability of the materials in acidic and basic media in multiphase extraction. Those skilled in the art will apply the technique that is most likely to achieve the desired separation.

[0100] example The examples are intended to illustrate the invention without limiting it.

[0101] The chemical reactions described in the examples can be readily adapted to prepare many other inhibitors of the present invention, and alternative methods for preparing the compounds of the present invention are considered to be within the scope of the invention. For example, the synthesis of compounds not exemplified by the present invention may be successfully carried out by modifications obvious to those skilled in the art, for example, by appropriately protecting interfering groups, by using other suitable reagents known in the art other than those described, and / or by routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as applicable to preparing other compounds of the present invention.

[0102] Reagents were purchased from Acros Organics, Sigma-Aldrich, Apollo Scientific, or Fluorochem at the highest commercial quality and used without further purification. Solvents were purchased from Acros Organics in AcroSeal® bottles for molecular sieving. Grignard, cross-coupling, and peptide coupling reactions were carried out in anhydrous solvents under a nitrogen atmosphere, and glassware was oven-dried before use. Thin-layer chromatography (TLC) plates were purchased from Merck KGaA (Polygram SIL / UV254, 0.2 mm silica with fluorescent indicator), and compounds were visualized using UV light (254 nm). Flash chromatography was performed using an Isco CombiFlash Companion system with a packed silica gel column (RediSep with a particle size of 40-60 μm). 1 H, 19 F and 13 The NMR spectrum of ¹³C was recorded using a Bruker Avance 400 spectrometer. The NMR spectrum was obtained in a deuterated solvent, i.e., CDCl3 or (CD3)2SO. The chemical shift (δ value) is reported in ppm, and for CDCl3, the deuterated solvent (7.26 ppm) was used. 1 (H NMR) and 77.16 ppm ( 13 ¹¹¹ NMR); and for (CD3)2SO, 2.50 ppm (1 (H NMR) and 39.52 ppm ( 13 The signal is corrected to that of 13C NMR. 19 The 1F NMR spectrum is calibrated against CFCl3 (δ=0 ppm) as an external standard. When peak multiplicity is reported, the following abbreviations are used: s (singular), d (double), dd (double of double), t (tripular), td (triple of double), q (quadruplicate), quadruplicate double (dq), m (multiplex), br (broadening signal). Coupling constants are reported in Hertz (Hz) if given. High-resolution mass spectra (HRMS) were recorded using a Thermo Fisher Scientific LTQ Orbitrap XL (nanoESI-MS) spectrometer. MALDI-ToF mass spectra were obtained using a Voyager-De® Pro, measured in m / z. The chromatographic purity of the final compound was determined by high-performance liquid chromatography (HPLC) analysis using a ThermoFisher Ultimate 3000SD system equipped with an LPG-3400SD pump system, ACC-3000 autosampler and column oven, and a DAD-3000 diode array detector. A ThermoFisher Acclaim-120 C18 reversed-phase column was used as the stationary phase. CH3CN / MeOH:H2O (10:90) A mobile phase consisting of [components] was subjected to gradient elution (5:95 for 0.2 minutes, 5:95 to 100:0 for 10 minutes, and 100:0 for 3 minutes) at 40°C with a flow rate of 0.5 mL / min. The purity of all final compounds was higher than 95%.

[0103] The following abbreviations will be used below: DMSO (dimethyl sulfoxide), HCl (hydrochloric acid), M (molar concentration), MALDI (matrix-assisted laser desorption / ionization), HRMS (high-resolution mass spectrometry), MS (mass spectrometry), PBS (phosphate-buffered saline), TLC (thin-layer chromatography).

[0104] Preparation of intermediate compounds: The intermediate compounds used to produce the compounds of formula (I, II) were prepared using the following method.

[0105] Method 1: tert-butyl 4-(4-chloro-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-carboxylate [ka] To a solution of 2,4-dichloro-6-(morpholine-4-yl)-1,3,5-triazine (20.7 g, 87.9 mmol, 0.9 equivalents) in ethanol, DIPEA (16.2 g, 21.4 ml, 125.6 mmol, 1.3 equivalents) and 1-Boc-piperazine (18.0 g, 96.6 mmol, 1.0 equivalent) were added at 0°C. The reaction mixture was stirred at room temperature for 5 hours. The solvent was removed under reduced pressure. Dichloromethane (300 mL) was added, and the resulting organic layer was washed with saturated NaHSO4 aqueous solution (4 × 200 mL). The organic layer was dehydrated with anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The product was recrystallized from dichloromethane / heptane to obtain tert-butyl 4-(4-chloro-6-morpholino-1,3,5-triazin-2-yl)piperazine-1-carboxylate as a colorless solid (22.7 g, 58.9 mmol, 80%). MALDI-MS: m / z = 385.643 [M + H] + HPLC:t R =6.53 minutes (purity 100.0%).

[0106] Method 2: 4-(difluoromethyl)-5-(4-morpholino-6-(piperazin-1-yl)-1,3,5-triazin-2-yl)pyrimidine-2-amine [ka] Step 1. Tert-butyl N-[5-bromo-4-(difluoromethyl)pyrimidine-2-yl]-N-[(tert-butoxy)carbonyl]carbamate (7.00 g, 16.50 mmol, 1.0 equivalent), bis(pinacolate)diborone (6.29 g, 24.75 mmol, 1.5 equivalents), potassium acetate (5.02 g, 51.15 mmol, 3.1 equivalents), and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (Pd(dppf)Cl2, 1.21 g, 1.65 mmol, 0.1 equivalent) were placed in a flask under a nitrogen atmosphere. Anhydrous 1,4-dioxane (40 mL) was added, and the mixture was stirred at 95°C for 1.5 hours. After the reaction was complete, the mixture was cooled to room temperature. Step 2: Monochlorotriazine 4-(4-chloro-6-morpholino-1,3,5-triazin-2-yl)piperazine-1-carboxylate (6.99 g, 18.15 mmol, 1.1 equivalents), chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (XPhos Pd G2, 0.519 g, 0.66 mmol, 0.04 equivalents), tripotassium phosphate (10.51 g, 49.50 mmol, 3.0 equivalents), and deionized H2O (10 mL) were added. The resulting mixture was stirred overnight at 100°C. After the reaction was complete, the mixture was allowed to cool to room temperature, and the crude product was filtered through Celite. Deionized H2O (300 mL) and ethyl acetate (300 mL) were added to the solution. The layers were separated, and the organic layer was washed with deionized H2O and brine (twice). The aqueous layer was extracted with dichloromethane (twice). The combined organic layers were concentrated under reduced pressure. Step 3. The above residue was dissolved in 1,4-dioxane (40 mL), and aqueous HCl (3 M, 40 mL) was added. The reaction mixture was stirred overnight at 80°C. After the reaction was complete, the mixture was cooled to room temperature. Ethyl acetate (200 mL) and deionized H2O (200 mL) were added, and the two layers were separated. The aqueous layer was washed with ethyl acetate (three times). The aqueous layer was basicized to pH=10.The formed solid was filtered and washed with acetonitrile to obtain 4-(difluoromethyl)-5-(4-morpholino-6-(piperazin-1-yl)-1,3,5-triazin-2-yl)pyrimidine-2-amine as a beige solid (4.91 g, 12.48 mmol, 76%). MALDI-MS: m / z = 394.205 [M+H]. + .

[0107] Method 3: tert-butyl3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)(methyl)carbamoyl)azetidine-1-carboxylate [ka] To a solution of 1-(tert-butoxycarbonyl)azetidine-3-carboxylic acid (581 mg, 2.89 mmol, 1.2 equivalents) in anhydrous N,N-dimethylformamide (DMF, approximately 1 mL / 0.18 mmol), O-(1H-6-chlorobenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU, 1.2 equivalents) and N,N-diisopropylethylamine (3.2 equivalents) were added at 0°C under a nitrogen atmosphere. The resulting mixture was stirred at 0°C for 5 minutes, then 2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-N-methyl-2-oxoethane-1-aminium chloride (or their respective HCl salts) were added, and the reaction mixture was stirred at room temperature for 4 to 16 hours. After the reaction was complete, DMF was removed under high vacuum. The crude product was dissolved in DCM, and the organic layer was washed with deionized H2O (twice) and saturated Na2CO3 aqueous solution (three times). The organic layer was dehydrated with Na2SO4, filtered, and concentrated under reduced pressure. Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 96:4:0.04) yielded the compound tert-butyl 3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-2-oxoethyl)(methyl)carbamoyl)azetidine-1-carboxylate as a colorless solid (1.237 g, 1.91 mmol, 79%). MALDI-MS: m / z = 648.7 [M+H] + m / z = 548.4[(Mt-butyl)+H] + .

[0108] Method 4: 3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-2-oxoethyl)(methyl)carbamoyl)azetidine-1-ium chloride [ka] To a solution of each Boc-protecting amine (1.0 equivalent) in THF (approximately 1 mL / 0.10 mmol), a 4 M solution of HCl (17 equivalents) in dioxane was added dropwise. The mixture was stirred at room temperature for 16 hours. The mixture was dried under reduced pressure. The product was precipitated from ACN, filtered, and washed with cold ACN. The HCl salt of the product was used in the next step without further purification. 3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)(methyl)carbamoyl)azetidine-1-ium chloride was obtained as a colorless solid (987 mg, 1.69 mmol, 91%). MALDI-MS: m / z = 548.4 [M + H] + .

[0109] Preparation of the compound of the present invention General procedure 1: To a solution of each carboxylic acid (1.0-1.5 equivalents) in anhydrous N,N-dimethylformamide (DMF, approximately 1 mL / 0.18 mmol), O-(1H-6-chlorobenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HCTU, 1.1 equivalents) and N,N-diisopropylethylamine (3.2 equivalents) were added under a nitrogen atmosphere at 0°C. The resulting mixture was stirred at 0°C for 5 minutes, then the respective HCl salts (1.0 equivalent) were added, and the reaction was stirred at room temperature for 4-16 hours. After the reaction was complete, the DMF was removed under high vacuum. The crude product was dissolved in DCM, and the organic layer was washed with deionized H2O (twice) and saturated aqueous Na2CO3 (three times). The organic layer was dehydrated with Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography and then recrystallized from dichloromethane / pentane.

[0110] Example 1: 1-((R)-3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-yl)propa-2-en-1-one [ka] 1-((R)-3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-yl)propa-2-en-1-one, according to general procedure 1, (R)-3-((2-(4-(4- The compound was prepared from (2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-ium chloride (193 mg, 0.32 mmol, 1.0 equivalent) and acrylic acid (26 mg, 0.35 mmol, 1.1 equivalent). Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 96:4:0.04) yielded the desired compound as a colorless solid (65 mg, 0.11 mmol, 33%). HRMS (m / z): [M+Na]+C 28 H 38 F2N 10 Calculated value for NaO4: 639.2938; Measured value: 639.2948. HPLC (acetonitrile containing 0.1% TFA): t R =7.20 minutes (98.9% purity).

[0111] Example 2: 1-((R)-3-(3-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5-triazine-2-yl)piperazine-1-yl)-3-oxopropoxy)piperidine-1-yl)propa-2-en-1-one [ka] 1-((R)-3-(3-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5triazine-2-yl)piperazine-1-yl)-3-oxopropoxy)piperidine-1-yl)propa-2-en-1-one, according to general procedure 1, (R)-3-(3-(4-(4-(2 The compound was prepared from -amino-4-(difluoromethyl)pyrimidine-5-yl)-6-((S)-3-methylmorpholino)-1,3,5-triazine-2-yl)piperazine-1-yl)-3-oxopropoxy)piperidine-1-ium chloride (150 mg, 0.25 mmol, 1.0 equivalent) and acrylic acid (20 mg, 0.27 mmol, 1.1 equivalent). Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 97:3:0.03) yielded the desired compound as a colorless solid (32 mg, 0.052 mmol, 20%). HRMS (m / z): [M+Na]+C 28 H 38 F2N 10 Calculated value for NaO4: 639.2938; Measured value: 639.2947. HPLC: t R =7.08 min (purity >99.9%).

[0112] Example 3: (R,Z)-2-(3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-carbonyl)-3-cyclopropylacrylonitrile [ka] (R,Z)-2-(3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-carbonyl)-3-cyclopropylacrylonitrile according to general procedure 1, (R)-3-((2-(4-(4-(2-amino The compound was prepared from (226 mg, 0.39 mmol, 1.0 equivalent) no-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethoxy)methyl)piperidine-1-ium chloride and (Z)-2-cyano-3-cyclopropylacrylic acid (118 mg, 0.85 mmol, 2.2 equivalents). Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 92:8:0.08) yielded the desired compound as a colorless solid (148 mg, 0.22 mmol, 57%). HRMS (m / z): [M+Na]+C 31 H 39 F2N 11 Calculated value for NaO4: 690.3047; Measured value: 690.3052. HPLC: t R =7.50 minutes (96.7% purity).

[0113] Example 4: (R)-1-Acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)-N-methylpiperidine-3-carboxamide [ka] (R)-1-acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)-N-methylpiperidine-3-carboxamide was prepared according to General Procedure 1 from (R)-3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)(methyl)carbamoyl)piperidine-1-ium chloride (1.139 g, 1.86 mmol, 1.0 equivalent) and acrylic acid (147 mg, 2.05 mmol, 1.1 equivalent). Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 97:3:0.03) yielded the desired compound as a colorless solid (305 mg, 0.48 mmol, 26%). HRMS (m / z): [M+Na]+C 28 H 37 F2N 11 Calculated value for NaO4: 652.2890; Measured value: 652.2893. HPLC: t R =6.20 minutes (98.3% purity).

[0114] Example 5: (R)-1-Acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-2-oxoethyl)pyrrolidine-3-carboxamide [ka] (R)-1-acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-2-oxoethyl)pyrrolidine-3-carboxamide was prepared from (R)-3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazin-1-yl)-2-oxoethyl)carbamoyl)pyrrolidine-1-ium chloride (274 mg, 0.47 mmol, 1.0 equivalent) and acrylic acid (37 mg, 0.52 mmol, 1.1 equivalent) according to General Procedure 1. Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 96:4:0.04) yielded the desired compound as a colorless solid (134 mg, 0.22 mmol, 47%). HRMS(m / z):[M+Na]+C 26 H 33 F2N 11 Calculated value for NaO4: 624.2577; Measured value: 624.2581. HPLC: t R =5.84 minutes (98.4% purity).

[0115] Example 6: 1-Acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)-N-methylazetidine-3-carboxamide [ka] 1-Acryloyl-N-(2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)-N-methylazetidine-3-carboxamide was prepared from 3-((2-(4-(4-(2-amino-4-(difluoromethyl)pyrimidine-5-yl)-6-morpholino-1,3,5-triazine-2-yl)piperazine-1-yl)-2-oxoethyl)(methyl)carbamoyl)azetidine-1-ium chloride (946 mg, 1.62 mmol, 1.0 equivalent) and acrylic acid (128 mg, 1.78 mmol, 1.1 equivalent) according to General Procedure 1. Purification by silica gel column chromatography (dichloromethane / methanol / ammonia: 100:0:0 → 96:4:0.04) yielded the desired compound as a colorless solid (358 mg, 0.60 mmol, 37%). HRMS(m / z):[M+Na]+C 26 H 33 F2N 11 Calculated value for NaO4: 624.2577; Measured value: 624.2579. HPLC: t R =5.81 minutes (98.4% purity).

[0116] Determination of the apparent dissociation constant of the inhibitor. The apparent dissociation constant of the compound for p110α [K i(app) ] References 21 Determined by LanthaScreen Technology (Life Technologies) as described in [document / document].

[0117] Determination of the velocity constant The maximum potential rate (k) of covalent bond formation in the compound for p110α inact ) and the dissociation constant (K) of the first reversible bond i The parameters were determined by LanthaScreen Technology (Life Technologies). Dynamic parameters were calculated using KinTek Global Kinetic Explorer modeling software. 22~27This was done by global fitting for numerical integration using [a specific method / tool].

[0118] In-Cell Western Cellular PI3K Signaling and IC50 50 decision Protein phosphorylation was detected as follows: Using a rabbit polyclonal antibody (number 4058) from Cell Signaling Technology (CST), pSer473 of PKB / Akt was detected by in-cell Western assay. In this case, 1.2 × 10⁶ wells were detected per well in a 96-well plate. 4 Individual SKOV3 cells, reference 21 As described, the cells were plated for 24 hours (37°C, 5% CO2) (Cell Carrier, PerkinElmer).

[0119] NanoBRET Target Binding Assay The N-terminal NanoLuc-fused PI3K was encoded in a pFN31K expression vector (Promega) containing a flexible Gly-Ser-Ser-Gly-Ala-Ile-Ala linker between NanoLuc and full-length target kinases including PI3Kα, PI3Kα C862S, PI3Kβ, and PI3Kδ. HEK293 cells were co-transfected with NanoLuc / PI3K and its regulatory subunit p85 in a mass ratio of 1:10 using the jetPEI transfection reagent (Polyplus transfection, no. 101B-010N). Table 1. Intrinsic reactivity and in vitro data of some of the compounds of the present invention and CNX-1351. [Table 9-1] [Table 9-2] Table 2. Cellular data for some of the compounds of the present invention and CNX-1351. Table 10

[0120] References JPEG0007870088000047.jpg102164

Claims

1. Compound of formula (IV) 【Chemistry 1】 [In the formula, X is CH or N, Y is either H or F, ・R １ and R ２ H and CH are independent of each other. ３ cyclopropyl, -F, -CH ２ -F, -CH ２ -CH ２ -F, -CN, 【Chemistry 2】 (wherein, R ５ is F or CH ３ and R ６ is C １～６ alkyl, and z is 0, 1 or 2) and is selected from ・R ３ is C １～３ It is either an alkyl group or two residues R ３ is bridged - (CH ２ ) ｒ - forms, where r is 1, 2 or 3, v is 0, 1, 2, 3 or 4, ・R ４ is H, F, or -CN, ・L ２ teeth, 【Transformation 3】 (In the formula, R ５ is C １～３ Alkyl, F, -CH ２ The portion is selected from CN or -CN, where t is 0, 1 or 2. ・W １ is CO or CH ２ And, ・W ２ is O, CH ２ and selected from CO, U is O, CH ２ , CO, NH, and N(CH ３ ) are selected from, n is either 1 or 2] or its tautomers, solvates, or pharmaceutically acceptable salts.

2. Compound of formula (IVa) 【Chemistry 4】 [In the formula, R1 and R2 are independently H, CH3, cyclopropyl, -F, -CH2-F, -CH2-CH2-F, -CN 【Transformation 5】 (wherein R 5 is F or CH 3, R 6 is C 1-6 alkyl, and z is 0, 1 or 2) R3 is C1-3 alkyl, or two residues R3 form a cross-linking -(CH2)r- where r is 1, 2, or 3. v is 0, 1, 2, 3, or 4. R4 is H, F, or -CN. L2 is, 【Transformation 6】 (wherein R5 is a part selected from C1-3 alkyl, F, -CH2CN or -CN, and t is 0, 1 or 2) W1 is CO or CH2, W2 is selected from O, CH2, and CO. U is selected from O, CH₂, CO, NH, and N(CH₃), n is either 1 or 2] or its tautomers, solvates, or pharmaceutically acceptable salts.

3. R １ However, H, CH ３ or -CH ２ The compound according to claim 1 or 2, wherein F.

4. R ２ The compound according to any one of claims 1 to 3, wherein the compound is H or cyclopropyl.

5. R ２ is cyclopropyl, and R ４ The compound according to any one of claims 1 to 4, wherein is -CN.

6. R ３ However, C １～３ The compound according to any one of claims 1 to 5, wherein it is alkyl.

7. The compound according to any one of claims 1 to 6, wherein v is 0, 1, or 2.

8. U is O, CH ２ NH and N(CH ３ A compound according to any one of claims 1 to 7, selected from ).

9. Compound of formula (V) 【Transformation 7】 (In the formula, X is CH or N, Y is either H or F, R1 and R2 are independently H, CH3, cyclopropyl, -F, -CH2-F, -CH2-CH2-F, -CN 【Transformation 8】 (wherein R 5 is F or CH 3, R 6 is C 1-6 alkyl, and z is 0, 1 or 2) R3 is C1-3 alkyl, or two residues R3 form a cross-linking -(CH2)r- where r is 1, 2, or 3. R4 is H, F, or -CN. L2 is, 【Chemistry 9】 (wherein R5 is a part selected from C1-3 alkyl, F, -CH2CN or -CN, and t is 0, 1 or 2) W1 is CO or CH2, W2 is selected from O, CH2, and CO. U is selected from O, CH₂, CO, NH, and N(CH₃), n is either 1 or 2, v is either 0 or 1. The compound according to any one of claims 1 to 8.

10. Compound of formula (Va) 【Chemistry 10】 [In the formula, R1 and R2 are independently H, CH3, cyclopropyl, -F, -CH2-F, -CH2-CH2-F, -CN 【Chemistry 11】 (wherein R 5 is F or CH 3, R 6 is C 1-6 alkyl, and z is 0, 1 or 2) R3 is C1-3 alkyl, or two residues R3 form a cross-linking -(CH2)r- where r is 1, 2, or 3. R4 is H, F, or -CN. L2 is, 【Chemistry 12】 (wherein R5 is a part selected from C1-3 alkyl, F, -CH2CN or -CN, and t is 0, 1 or 2) W1 is CO or CH2, W2 is selected from O, CH2, and CO. U is selected from O, CH₂, CO, NH, and N(CH₃), n is either 1 or 2, - v is either 0 or 1] The compound according to any one of claims 1 to 9.

11. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10, for use in the treatment of a disease.

12. The pharmaceutical composition according to claim 11, wherein the disease is caused by an activating mutation in the PI3KCA gene or activation of class I PI3K.

13. A pharmaceutical composition comprising a compound according to any one of claims 1 to 10 for use in the treatment of tumor diseases, overgrowth syndromes, neurological disorders, and immune disorders.

14. The pharmaceutical composition according to claim 13, wherein the tumor is a solid tumor.

15. The pharmaceutical composition according to claim 13, wherein the tumor disease is selected from lymphoma and leukemia.

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