CN109721531B - Novel liposome kinase inhibitor - Google Patents
Novel liposome kinase inhibitor Download PDFInfo
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- CN109721531B CN109721531B CN201711058134.9A CN201711058134A CN109721531B CN 109721531 B CN109721531 B CN 109721531B CN 201711058134 A CN201711058134 A CN 201711058134A CN 109721531 B CN109721531 B CN 109721531B
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract
The present invention provides a novel kinase inhibitor comprising a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof. The invention also provides pharmaceutical compositions comprising compounds of formula (I) and uses and methods of using these compounds and compositions for treating conditions mediated by PI3K kinase or PfPI4K kinase activation, particularly malaria, cancer, and other cell proliferative diseases.
Description
Technical Field
The present application relates to a novel class of kinase inhibitor compounds, pharmaceutical compositions comprising these compounds, and uses and methods of using these compounds and compositions for treating conditions mediated by PI3K kinase or PfPI4K kinase activation, particularly malaria, cancer, and other cell proliferative diseases.
Background
Many viruses affect host lipid metabolism, lipid membrane trafficking, and lipid-mediated signal transduction. Phosphoinositides (PI) are a class of phospholipids that are involved in several cellular processes. Phosphatidylinositol is the basic skeleton of PI, and 7 types of PIs are obtained in total after the 3,4, 5-hydroxyl groups of 5 hydroxyl groups of phosphatidylinositol are reversibly phosphorylated by kinases, including phosphatidylinositol-3-phosphate (phosphatidylinositol 3-phosphate, PI3P), phosphatidylinositol-4-phosphate (phosphatidylinositol 4-phosphate, PI4P), phosphatidylinositol-5-phosphate (phosphatidylinositol 5-phosphate, PI5P), phosphatidylinositol-3, 4-diphosphate (phosphatidylinositol 3, 4-diphosphate, PI (3,4) P2), phosphatidylinositol-3, 5-diphosphate (phosphatidylinositol 3,5) P2), phosphatidylinositol-4, 5-diphosphate (phosphatidylinositol 4, 5-diphosphate) P8628, phosphatidylinositol-4, 5-phosphate (phosphatidylinositol 4, 5-phosphate, PI3, 5-diphosphate) P2, and phosphatidylinositol-4, 5-phosphate (phosphatidylinositol 3, 5-phosphate, 864, 2,3, 864-diphosphate, 5-triphosphate (phosphatydilisinositol 3,4,5-triphosphate, PI (3,4,5) P3). These 7 PIs can be switched over with each other, and different types of phosphatases and kinases participate in their switching over process. The distribution of these PIs in subcellular membranes varies. PI3P was distributed throughout the morning and evening, PI4P was distributed mainly in the golgi apparatus; PI (3,4) P2, PI (4,5) P2, PI (3,4,5) P3 are mainly located in plasma membrane; PI (3,5) P2 was distributed mainly in synaptic secretory vesicles, with a minor distribution in late endosomes.
PIs are a negatively charged phospholipid that is widely present in cell membranes and, although present in low levels in biological membranes, are an important component of biological membranes. It plays an important role in membrane permeability, transport of vesicles, membrane translocation, cytoskeletal regulation, and signaling pathways.
Recent studies have found that phosphatidylinositol-4-kinase (PI 4K) is a major target for plasmodium falciparum and that malaria treatment can be achieved by selectively inhibiting the PI4K protein of plasmodium falciparum, but not inhibiting the human PI4K protein (Nature,2013, 504, 248-253).
The present patent mainly relates to a novel kinase inhibitor, the action targets of which are mainly Plasmodium falciparum PI4K (Plasmodium falciparum phosphatydicolinositol 4-kinase, PfPI4K) and phosphatidylinositol-3-kinase (phosphatydicolinol 3-kinases, PI3K), but there is no obvious inhibition effect on human PI4K, thereby achieving the effect of treating malaria. The compounds of the present patent also have significant inhibitory effects on PI3K protein and thus can be used to treat disorders mediated by PI 3K.
Disclosure of Invention
The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof:
wherein:
x is selected from CH or N;
ring A is selected from the group consisting of pyridyl, pyrrolyl, pyrrolopyridyl, pyrazolyl, pyrazolopyridyl, tetrahydropyridinyl, isoquinolinyl, phenyl, and indolyl, said ring A having its nitrogen atom optionally substituted with an amino protecting group;
R 1 and R 2 Each independently selected from H, C 1-6 Alkyl, halogen, and C 1-6 A haloalkyl group;
R 3 selected from H, halogen, C 1-6 Alkyl, and C 1-6 An alkoxy group;
R 4 selected from H, C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Haloalkoxy, C 1-6 Alkanoyl radical, C 1-6 Alkylaminocarbonyl radical, C 3-6 Cycloalkylaminocarbonyl group, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, phenylalkyl, phenylaminocarbonyl, phenylalkylaminocarbonyl, pyridinylaminocarbonyl, and amino protecting groups, which are optionally substituted with amino, amino protecting group-substituted amino, hydroxy or halogen.
In a preferred aspect, the A ring is selected from the group consisting of pyridin-3-yl, pyridin-4-yl, pyrrol-3-yl, pyrrolo [2,3-b ] pyridin-5-yl, pyrazol-4-yl, pyrazolo [3,4-b ] pyridin-5-yl, tetrahydropyridin-4-yl, isoquinolin-4-yl, phenyl, and indol-2-yl, and the nitrogen atom of the A ring is optionally substituted with an amino protecting group selected from the group consisting of pivaloyl, t-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, and trifluoroacetyl.
Further preferably, R 1 And R 2 At least one of which is H.
Further preferably, R 3 Selected from the group consisting of chlorine, methyl, and methoxy.
In a preferred embodiment, R 4 Is selected from C 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, phenylaminocarbonyl, phenylalkylaminocarbonyl, and pyridylaminocarbonyl, the above substituents being optionally substituted with amino, amino protecting group-substituted amino, hydroxy or halogen. Further preferably, R 4 Selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, acetamido, dimethylaminoacetamido, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl, and 2-pyridylaminocarbonyl.
In yet another aspect, the present invention relates to a compound of formula (II) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof:
wherein:
x is selected from CH or N;
R 1 and R 2 At least one of which is H and the other is selected from H, C 1-6 Alkyl, halogen, and C 1-6 A haloalkyl group;
R 3 selected from H, halogen, C 1-6 Alkyl, and C 1-6 An alkoxy group;
R 4 selected from H, C optionally substituted by amino 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, phenylaminocarbonyl optionally substituted with halogen, phenylalkylaminocarbonyl, and pyridylaminocarbonyl.
Preferably, R 1 And R 2 At least one of which is H and the other is selected from H, methyl, 2-propyl, fluoro, chloro, and trifluoromethyl.
Further preferably, R 3 Selected from the group consisting of chlorine, methyl, and methoxy.
In other embodiments, R 4 Selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, acetamido, dimethylaminoacetamido, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl, and 2-pyridylaminocarbonyl.
In another aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of at least one compound provided herein, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.
In another aspect, the present application relates to methods and uses for inhibiting PI3K and/or PfPI4K activity comprising administering a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, or a pharmaceutical composition comprising a compound of the present invention.
In another aspect, the present application relates to methods and uses for treating, preventing or ameliorating a disease mediated by PI3K and/or PfPI4K, in particular for treating, preventing or ameliorating malaria, comprising administering to a subject a compound of the present invention, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, or a pharmaceutical composition comprising a compound of the present invention.
Detailed Description
Term(s) for
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs.
The present invention employs, unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, within the skill of the art. Unless a specific definition is provided, nomenclature and laboratory procedures and techniques related to the chemistry described herein, such as analytical chemistry, synthetic organic chemistry, and medical and pharmaceutical chemistry, are known to those skilled in the art. In general, the foregoing techniques and procedures may be practiced by conventional methods well known in the art and described in various general and more specific documents that are cited and discussed in this specification.
The term "alkyl" refers to an aliphatic hydrocarbon group, which may be a branched or straight chain alkyl group. Depending on the structure, the alkyl group may be a monovalent group or a divalent group (i.e., alkylene). In the present invention, the alkyl group is preferably an alkyl group having 1 to 8 carbon atoms, more preferably a "lower alkyl group" having 1 to 6 carbon atoms, and even more preferably an alkyl group having 1 to 4 carbon atoms. Typical alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, and the like. It is to be understood that reference herein to "alkyl" includes reference to that alkyl in all configurations and conformations that may exist, for example reference herein to "propyl" includes n-propyl and isopropyl, "butyl" includes n-butyl, isobutyl and tert-butyl, "pentyl" includes n-pentyl, isopropyl, neopentyl, tert-pentyl, and pent-3-yl, and the like.
The term "alkoxy" refers to an-O-alkyl group, wherein alkyl is as defined herein. Typical alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, butoxy, pentoxy, hexoxy, and the like.
The term "cycloalkyl" refers to a monocyclic or multicyclic group that contains only carbon and hydrogen. Cycloalkyl groups include groups having 3 to 12 ring atoms. Depending on the structure, the cycloalkyl group can be a monovalent group or a divalent group (e.g., cycloalkylene). In the present invention, the cycloalkyl group is preferably a cycloalkyl group having 3 to 8 carbon atoms, more preferably a "lower cycloalkyl group" having 3 to 6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, and adamantyl.
The term "aryl" refers to a planar ring having a delocalized pi-electron system and containing 4n +2 pi electrons, where n is an integer. An aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. The aryl group may be optionally substituted. The term "aryl" includes carbocyclic aryl (e.g., phenyl) and heterocyclic aryl (or "heteroaryl") groups (e.g., pyridine). The term includes monocyclic or fused-ring polycyclic (i.e., rings that share adjacent pairs of carbon atoms) groups.
The term "aryl" as used herein means that each of the ring-forming atoms in the aromatic ring is a carbon atom. The aryl ring may be composed of five, six, seven, eight, nine or more than nine atoms. The aryl group may be optionally substituted. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, phenanthryl, anthracyl, fluorenyl, and indenyl. Depending on the structure, the aryl group can be a monovalent group or a divalent group (i.e., arylene).
The term "heteroaryl" refers to an aryl group that includes one or more ring heteroatoms selected from nitrogen, oxygen, and sulfur. An N-containing "heteroaryl" moiety means that at least one of the backbone atoms in the ring in the aryl group is a nitrogen atom. Depending on the structure, heteroaryl groups can be monovalent or divalent (i.e., heteroarylene). Examples of heteroaryl groups include, but are not limited to, pyridyl, imidazolyl, pyrimidinyl, pyrazolyl, triazolyl, pyrazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, thiazolyl, oxazolyl, isothiazolyl, pyrrolyl, quinolyl, isoquinolyl, indolyl, benzimidazolyl, benzofuranyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothienyl, benzothiazolyl, benzoxazolyl, quinazolinyl, naphthyridinyl, and furopyridyl, and the like.
The term "alkyl (aryl)" or "aralkyl" means that an alkyl group, as defined herein, is substituted with an aryl group, as defined herein. Non-limiting alkyl (aryl) groups include benzyl, phenethyl, and the like.
The term "heteroalkyl," as used herein, means that one or more backbone chain atoms in an alkyl group, as defined herein, is a heteroatom, such as oxygen, nitrogen, sulfur, silicon, phosphorus, or combinations thereof. The heteroatom(s) may be located anywhere within the heteroalkyl group or at the position where the heteroalkyl group is attached to the remainder of the molecule.
The term "heterocycloalkyl" or "heterocyclyl" as used herein means that one or more of the ring-forming atoms in the non-aromatic ring is a heteroatom selected from nitrogen, oxygen and sulfur. A heterocycloalkyl ring can be composed of three, four, five, six, seven, eight, nine, or more than nine atoms. The heterocycloalkyl ring may be optionally substituted. Examples of heterocycloalkyl groups include, but are not limited to, lactams, lactones, cyclic imines, cyclic thioimines, cyclic carbamates, tetrahydrothiopyrans, 4H-pyrans, tetrahydropyrans, piperidines, 1, 3-dioxins, 1, 3-dioxanes, 1, 4-dioxins, 1, 4-dioxanes, piperazines, 1, 3-oxathianes, 1, 4-oxathianes, tetrahydro-1, 4-thiazines, 2H-1, 2-oxazines, maleimides, succinimides, barbituric acid, thiobarbituric acid, dioxopiperazines, hydantoins, dihydrouracils, morpholines, trioxanes, hexahydro-1, 3, 5-triazines, tetrahydrothiophenes, tetrahydrofurans, pyrrolines, pyrrolidines, imidazolidines, pyrrolidones, 4-oxazides, 4-dioxanes, 1, 4-dioxanes, piperazines, 1, 4-oxazidines, 1, 4-oxazidines, 2H-oxazines, 2-oxazines, maleimides, succinimide, and pyrrolidine, Pyrazoline, pyrazolidine, imidazoline, imidazolidine, 1, 3-dioxole, 1, 3-dioxolane, 1, 3-dithiole, 1, 3-dithiolane, isoxazoline, isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazolidine, and 1, 3-oxathiolane. Depending on the structure, heterocycloalkyl groups can be monovalent or divalent (i.e., heterocycloalkylene).
The term "halo" or "halogen" refers to fluorine, chlorine, bromine and iodine.
The terms "haloalkyl", "haloalkoxy" and "haloheteroalkyl" include alkyl, alkoxy, or heteroalkyl groups in which at least one hydrogen is replaced with a halogen atom. In certain embodiments, if two or more hydrogen atoms are replaced with a halogen atom, the halogen atoms are the same or different from each other.
The term "amino" refers to the group-NH 2 。
The term "aminoacyl" refers to-CO-NH 2 。
The term "amido" or "amido" refers to the group-NR-CO-R ', wherein R and R' are each independently hydrogen or alkyl.
The term "alkylamino" refers to an amino substituent further substituted by one or two alkyl groups, in particular to the group-NRR ', wherein R and R ' are each independently selected from hydrogen or lower alkyl, with the proviso that-NRR ' is not-NH 2 . "alkylamino" includes the group-NH-thereof 2 The nitrogen of (a) is linked to at least one alkyl group. Examples of alkylamino groups include, but are not limited to, methylamino, ethylamino, and the like. "dialkylamino" includes wherein-NH 2 The nitrogen of (a) is linked to at least two other alkyl groups. Examples of dialkylamino groups include, but are not limited to, dimethylamino, diethylamino, and the like.
The terms "arylamino" and "diarylamino" refer to an amino substituent further substituted with one or two aryl groups, specifically the group-NRR ', where R and R' are each independently selected from hydrogen, lower alkyl, or aryl, where N is attached to at least one or two aryl groups, respectively.
The term "cycloalkylamino" refers to an amino substituent further substituted with one or two cycloalkyl groups as defined herein.
The term "aralkylamino" as used herein refers to the group-NRR 'wherein R is loweraralkyl and R' is hydrogen, loweralkyl, aryl or loweraralkyl.
The term "heteroarylamino" refers to an amino substituent further substituted with one or two heteroaryl groups as defined herein.
The term "alkylaminoalkyl" means that an alkyl group, as defined herein, is substituted with an alkylamino group, as defined herein.
The term "aminoalkyl" refers to an alkyl substituent further substituted with one or more amino groups.
The term "hydroxy" refers to a group of the formula-OH.
The term "hydroxyalkyl" or "hydroxyalkyl" refers to an alkyl substituent further substituted with one or more hydroxyl groups.
The term "acyl" refers to the monovalent radical remaining after removal of the hydroxyl group by an organic or inorganic oxoacid, and has the general formula R-M (O) -where M is typically C.
The term "carbonyl" is an organic functional group consisting of two atoms, carbon and oxygen, connected by a double bond (C ═ O).
The term "alkanoyl" or "alkylcarbonyl" refers to a carbonyl group further substituted with an alkyl group. Typical alkanoyl groups include, but are not limited to, acetyl, propionyl, butyryl, pentanoyl, hexanoyl, and the like.
The terms "alkylaminocarbonyl", "cycloalkylaminocarbonyl", "arylaminocarbonyl", "aralkylaminocarbonyl", "heteroarylaminocarbonyl", respectively, mean that the carbonyl group, as defined herein, is substituted with alkylamino, cycloalkylamino, arylamino, aralkylamino, or heteroarylamino, respectively, as defined herein.
The term "sulfonyl" or "sulfonyl" refers to a functional group of a sulfonic acid after loss of the hydroxyl group, specifically — S (═ O) 2 -a group.
The term "aminosulfonyl" or "aminosulfonyl" refers to-S (═ O) 2 -NH 2 A group.
Term(s) for"Alkylsulfonyl" or "alkylsulfonyl" refers to-S (═ O) 2 -R, wherein R is alkyl.
The term "optionally" means that one or more of the subsequently described events may or may not occur, and includes both occurring events and non-occurring events. The term "optionally substituted" or "substituted" means that the referenced group may be substituted with one or more additional groups each and independently selected from alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, hydroxy, alkoxy, cyano, halo, amido, nitro, haloalkyl, amino, methanesulfonyl, alkylcarbonyl, alkoxycarbonyl, heteroarylalkyl, heterocycloalkylalkyl, aminoacyl, amino protecting groups, and the like. Among them, the amino-protecting group is preferably selected from pivaloyl, t-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl, trifluoroacetyl and the like.
The term "inhibition", "inhibitory" or "inhibitor" of a kinase, as used herein, means that phosphotransferase activity is inhibited.
A "metabolite" of a compound disclosed herein is a derivative of the compound that is formed when the compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when the compound is metabolized. The term "metabolized" as used herein, refers to the sum of processes by which a particular substance is altered by an organism, including, but not limited to, hydrolysis reactions and reactions catalyzed by enzymes, such as oxidation reactions. Thus, enzymes can produce specific structural transformations into compounds. For example, cytochrome P450 catalyzes a variety of oxidation and reduction reactions, while phosphoglucose glycyltransferase catalyzes the conversion of activated glucuronic acid molecules to aromatic alcohols, aliphatic alcohols, carboxylic acids, amines and free sulfhydryl groups. Further information on metabolism can be obtained from The Pharmacological Basis of Therapeutics, ninth edition, McGraw-Hill (1996). Metabolites of the compounds disclosed herein can be identified by administering the compounds to a host and analyzing a tissue sample from the host, or by incubating the compounds with hepatocytes in vitro and analyzing the resulting compounds. Both methods are known in the art. In some embodiments, the metabolite of the compound is formed by an oxidation process and corresponds to the corresponding hydroxyl-containing compound. In some embodiments, the compound is metabolized to a pharmaceutically active metabolite. The term "modulate," as used herein, refers to interacting, directly or indirectly, with a target to alter the activity of the target, including by way of example only, enhancing the activity of the target, inhibiting the activity of the target, limiting the activity of the target, or prolonging the activity of the target.
IC as used herein 50 Refers to the amount, concentration or dose of a particular test compound that achieves 50% inhibition of the maximal effect in an assay that measures such effect.
GI as used herein 50 Refers to the concentration of drug required for 50% growth inhibition of the cells, i.e., the concentration of drug at which 50% of the cancer cells are inhibited or controlled by the drug.
Novel kinase inhibitors of the invention
The present invention relates to a compound of formula (I) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof:
wherein X is selected from CH or N;
the A ring is selected from the group consisting of pyridyl (e.g., pyridin-3-yl, pyridin-4-yl), pyrrolyl (e.g., pyrrol-3-yl), pyrrolopyridyl (e.g., pyrrolo [2,3-b ] pyridin-5-yl), pyrazolyl (e.g., pyrazol-4-yl), pyrazolopyridyl (e.g., pyrazolo [3,4-b ] pyridin-5-yl), tetrahydropyridinyl (e.g., tetrahydropyridin-4-yl), isoquinolinyl (e.g., isoquinolin-4-yl), aryl (e.g., phenyl), and indolyl (e.g., indol-2-yl), the nitrogen atom of which is optionally substituted with an amino protecting group, with the A ring being particularly preferably pyridin-3-yl;
R 1 and R 2 Each independently selected from H, C 1-6 Alkyl, halogen, and C 1-6 Haloalkyl, preferably R 1 And R 2 At least one is H, more preferably both are H;
R 3 selected from H, halogen, C 1-6 Alkyl, and C 1-6 Alkoxy, preferably selected from chloro, methyl, and methoxy;
R 4 selected from H, C 1-6 Alkyl radical, C 1-6 Alkoxy radical, C 1-6 Haloalkoxy, C 1-6 Alkanoyl radical, C 1-6 Alkylaminocarbonyl radical, C 3-6 Cycloalkylaminocarbonyl group, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, hydroxy C 1-6 Alkylaminocarbonyl, phenylalkyl, phenylaminocarbonyl, phenylalkylaminocarbonyl, pyridinylaminocarbonyl, and amino protecting groups, wherein amino is optionally substituted with an amino protecting group, said substituent being optionally substituted with amino, amino protecting group-substituted amino, hydroxy or halogen, R 4 Preferably selected from C 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, phenylaminocarbonyl, phenylalkylaminocarbonyl, and pyridylaminocarbonyl, more preferably selected from methylaminocarbonyl, ethylaminocarbonyl, acetylamino, dimethylaminoacetamino, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl, and 2-pyridylaminocarbonyl.
The amino protecting group is selected from pivaloyl, tert-butyloxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyl, p-methoxybenzyl, allyloxycarbonyl and trifluoroacetyl.
The present invention further relates to a compound of formula (II) or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof:
wherein X is selected from CH or N;
R 1 and R 2 At least one is H and the other is selected from H, C 1-6 Alkyl, halogen, and C 1-6 Haloalkyl, more preferably selected from H, methyl, 2-propyl, fluoro, chloro, and trifluoromethyl, most preferably R 1 And R 2 Are all H;
R 3 selected from H, halogen, C 1-6 Alkyl, and C 1-6 Alkoxy, preferably selected from chloro, methyl, and methoxy;
R 4 selected from H, C optionally substituted by amino 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, amino C 1-6 Alkylaminocarbonyl, phenylaminocarbonyl optionally substituted by halogen, phenylalkylaminocarbonyl, and pyridylaminocarbonyl, preferably selected from methylaminocarbonyl, ethylaminocarbonyl, acetylamino, dimethylaminoacetamino, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl, and 2-pyridylaminocarbonyl.
The invention relates to a chiral compound, the configuration of which can be any configuration or a mixed racemate.
The compounds described herein may be formulated and/or used as pharmaceutically acceptable salts. Types of pharmaceutically acceptable salts include, but are not limited to: (1) acid addition salts, formed by reacting the free base form of the compound with pharmaceutically acceptable inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, metaphosphoric acid, and the like; or with an organic acid such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, malic acid, citric acid, succinic acid, maleic acid, tartaric acid, fumaric acid, trifluoroacetic acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid, 4-methylbicyclo- [2.2.2] oct-2-ene-1-carboxylic acid, 2-naphthalenesulfonic acid, tert-butylacetic acid, glucoheptonic acid, 4' -methylenebis- (3-hydroxy-2-ene-1-carboxylic acid), 3-phenylpropionic acid, trimethylacetic acid, or mixtures thereof, Dodecyl sulfuric acid, gluconic acid, glutamic acid, salicylic acid, hydroxynaphthoic acid, stearic acid, muconic acid, and the like; (2) base addition salts, which are formed when an acidic proton in the parent compound is replaced by a metal ion, such as an alkali metal ion (e.g., lithium, sodium, potassium), alkaline earth metal ion (e.g., magnesium or calcium), or aluminum ion; or with an organic or inorganic base. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, trimethylamine, N-methylglucamine, and the like. Acceptable inorganic bases include aluminum hydroxide, calcium hydroxide, potassium hydroxide, sodium carbonate, sodium hydroxide, and the like.
The corresponding counterion of the pharmaceutically acceptable salt can be analyzed and identified using a variety of methods including, but not limited to, ion exchange chromatography, ion chromatography, capillary electrophoresis, inductively coupled plasma, atomic absorption spectroscopy, mass spectrometry, or any combination thereof.
Recovering the salt using at least one of the following techniques: filtration, precipitation with a non-solvent followed by filtration, solvent evaporation, or lyophilization are used in the case of aqueous solutions.
Screening and characterization of pharmaceutically acceptable salts, polymorphs, and/or solvates can be accomplished using a variety of techniques including, but not limited to, thermal analysis, X-ray diffraction, spectroscopy, microscopy, elemental analysis. Various spectroscopic techniques used include, but are not limited to, Raman, FTIR, UVIS, and NMR (liquid and solid state). Various microscopy techniques include, but are not limited to, IR microscopy and Raman (Raman) microscopy.
The invention relates to a pharmaceutical composition and application thereof
The present application provides pharmaceutical compositions formulated for administration by an appropriate route and manner, comprising an effective concentration of one or more compounds provided herein, or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug thereof, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.
The compounds of formula (I) or formula (II) in free form or in salt form, hereinafter also referred to as "agents of the invention", are useful for the treatment of diseases, disorders or conditions mediated by activation (including normal activity, especially over-activation) of one or more members of the PI3K kinase family, for example proliferative diseases, cancer, inflammatory or allergic diseases, obstructive airways diseases and/or conditions associated with transplantation, due to their inhibitory effect on phosphatidylinositol 3-kinase.
Due to the inhibitory effect of the "substances of the invention" in free form or in salt form on the phosphatidylinositol 4-kinase of plasmodium falciparum, the compounds of formula (I) or formula (II) in free form or in pharmaceutically acceptable salt form are useful for the treatment of diseases, disorders or conditions mediated by activation (including normal activity, especially over activation) of one or more members of the PfPI4K kinase family, such as malaria falciparum.
The "treatment" of the present invention may be therapeutic (e.g., symptomatic treatment) and/or prophylactic.
Preferably for use in the treatment of a proliferative disease selected from benign or malignant tumors, including but not limited to: brain cancer, kidney cancer, liver cancer, adrenal cancer, bladder cancer, breast cancer, lymphatic cancer, stomach cancer, gastric tumor, esophageal cancer, ovarian cancer, colorectal cancer, prostate cancer, pancreatic cancer, lung cancer, vaginal cancer, membrane adenocarcinoma, thyroid cancer, neck cancer, CNS cancer, glioblastoma, myeloproliferative disorders, sarcoma, glioblastoma, multiple myeloma, gastrointestinal cancer, head and neck tumor, brain tumor, epidermal hyperproliferation, psoriasis, prostate hyperplasia, neoplasia of epithelial character, lymphoma, or leukemia. Other diseases include Cowden syndrome (Cowden syndrome), Leelmite-Dudos disease and Bannayan-Zonana syndrome or diseases in which the PI3K/PKB pathway is abnormally activated. Preferably colorectal cancer, gastric cancer, breast cancer, lung cancer, liver cancer, prostate cancer, membrane adenocarcinoma, thyroid cancer, bladder cancer, kidney cancer, brain tumor, neck cancer, CNS cancer, glioblastoma, myeloproliferative disorders, leukemia and lymphoma.
The agents of the invention are useful in the treatment of inflammatory or obstructive airways diseases resulting in, for example, tissue damage, airways inflammation, bronchial hyperreactivity, remodelling or reduction in disease progression. Inflammatory or obstructive airways diseases to which the present invention is applicable include asthma of whatever type or genesis including both intrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mild asthma, moderate asthma, severe asthma, bronchitic asthma, exercise-induced asthma, occupational asthma and asthma induced following bacterial infection. Treatment of asthma is also understood to include treatment of individuals, for example individuals under the age of 4 or 5 years, who exhibit wheezing symptoms, diagnosed or diagnosable as "wheezy infant", a patient category of established major medical concern, now commonly identified as incipient or early asthma. For convenience, this particular asthma condition is known as "wheezy infant syndrome".
Prophylactic efficacy in the treatment of asthma will be manifested as a reduction in the frequency or severity of symptomatic attack, for example a reduction in the frequency or severity of acute asthmatic or bronchoconstrictive attack, an improvement in lung function or an improvement in airway hyperactivity. The efficacy is also manifested as a reduction in the need for treatment of other symptoms, i.e. treatments used or intended to limit or halt the onset of symptoms when they occur, such as anti-inflammatory drugs (e.g. corticosteroids) or bronchodilators. The prophylactic benefit for asthma may be particularly evident in individuals prone to "morning dipping". "morning dipping" is a recognized asthma syndrome, usually a significant proportion of asthma, characterized by attacks, for example, between about 4 and 6 am, i.e., generally at a time remote from any previously administered treatment for asthma symptoms.
Other inflammatory or obstructive airways diseases and conditions to which the present invention is applicable include Acute Lung Injury (ALI), adult/Acute Respiratory Distress Syndrome (ARDS), chronic obstructive pulmonary, airways or lung disease (COPD, COAD or COLD), including chronic bronchitis or dyspnea associated therewith, emphysema, and exacerbation of airways overactivity resulting from other drug therapy, particularly other inhaled drug therapy. The invention is also suitable for the treatment of bronchitis of any type or genesis including, for example, acute bronchitis, arachidic bronchitis, catarrhal bronchitis, fibrinous bronchitis, chronic bronchitis or tuberculous bronchitis. Other inflammatory or obstructive airways diseases to which the present invention is applicable include pneumoconiosis (an inflammatory, usually occupational, disease of the lungs, whether chronic or acute, with airways obstruction frequently encountered and resulting from repeated inhalation of dusts) of any type or genesis, including, for example, aluminosis, anthracosis, asbestosis, chalicosis, ptilosis, siderosis, silicosis, tabacosis and byssinosis.
The agents of the invention are also useful in the treatment of diseases, disorders or conditions mediated by phosphatidylinositol 3-kinase such as: respiratory diseases, allergies, rheumatoid arthritis, osteoarthritis, rheumatic disorders, psoriasis, ulcerative colitis, crohn's disease, septic shock, proliferative disorders, atherosclerosis, allograft rejection following transplantation, diabetes, stroke, obesity or restenosis, leukemia, interstitial tumors, thyroid cancer, systemic mastocytosis, hypereosinophilic syndrome, fibrosis, rheumatoid arthritis, polyarthritis, scleroderma, lupus erythematosus, graft-versus-host disease, neurofibromatosis, pulmonary hypertension, alzheimer's disease, seminoma, dysgerminoma, mast cell tumors, lung cancer, bronchial carcinoma, dysgerminoma, intraepithelial neoplasia, melanoma, breast cancer, neuroblastoma, papillary/follicular thyroid cancer, Malignant lymphoma, non-hodgkin's lymphoma, multiple endocrine neoplasia type 2, pheochromocytoma, thyroid cancer, parathyroid hyperplasia/adenoma, colon cancer, colorectal adenoma, ovarian cancer, prostate cancer, glioblastoma, brain tumor, malignant glioma, pancreatic cancer, malignant pleural mesothelioma, hemangioblastoma, hemangioma, renal cancer, liver cancer, adrenal cancer, bladder cancer, gastric cancer, rectal cancer, vaginal cancer, cervical cancer, endometrial cancer, multiple myeloma, neck and head tumors, neoplasia and other proliferative or proliferative diseases, or a combination thereof.
The substances of the invention are also useful for the treatment of eosinophil-related disorders, such as eosinophilia, in particular eosinophil-related airway disorders (e.g. lung tissue involving pathological eosinophil infiltration), including hypereosinophilia, as it affects the airway and/or lung, as well as eosinophil-related airway disorders, such as those caused by or associated with luxes syndrome, eosinophilic pneumonia, infestation by parasites (particularly metazoans), including tropical eosinophilia, bronchopulmonary aspergillosis, polyarteritis nodosa (including churg-strauss syndrome), eosinophil granuloma, and eosinophil-related disorders affecting the airway resulting from drug reactions.
The substances of the invention can also be used for the treatment of inflammatory or allergic conditions of the skin, such as psoriasis, contact dermatitis, atopic dermatitis, alopecia areata, erythema multiforme, dermatitis herpetiformis, scleroderma, vitiligo, allergic vasculitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphigus (pemphisus), epidermolysis bullosa acquisita and other inflammatory or allergic conditions of the skin.
The substances of the invention may also be used for the treatment of other diseases or conditions, in particular diseases or conditions having an inflammatory component, for example for the treatment of diseases and conditions of the eye, such as conjunctivitis, keratoconjunctivitis sicca and vernal conjunctivitis; diseases affecting the nose, including allergic rhinitis; and inflammatory diseases in which autoimmune reactions are involved or which have an autoimmune component or etiology, including autoimmune hematological disorders (e.g., hemolytic anemia, aplastic anemia, pure red cell anemia, and idiopathic thrombocytopenia), systemic lupus erythematosus, polychondritis, scleroderma, wegener's granulomatosis, dermatomyositis, chronic active hepatitis, myasthenia gravis, sjogren's syndrome, idiopathic sprue, autoimmune inflammatory bowel diseases (e.g., ulcerative colitis and Crohn's disease), endocrine ocular diseases, Graves' disease, sarcoidosis, alveolitis, chronic allergic pneumonia, multiple sclerosis, primary biliary cirrhosis, uveitis (anterior and posterior uveitis), keratoconjunctivitis sicca and vernal keratoconjunctivitis, interstitial pulmonary fibrosis, Psoriatic arthritis and glomerulonephritis (with and without nephrotic syndrome, including, for example, idiopathic nephrotic syndrome or minimal change nephropathy).
Other diseases or conditions which may be treated with the agents of the invention include septic shock, rheumatoid arthritis, osteoarthritis, proliferative diseases such as cancer, atherosclerosis, allograft rejection after transplantation, stroke, obesity, restenosis, diabetes, e.g. type I diabetes (juvenile diabetes) and type II diabetes, diarrheal diseases, ischemia/reperfusion injury, retinopathies such as diabetic retinopathy or hyperbaric oxygen induced retinopathy, and conditions characterised by elevated intraocular pressure or aqueous humor secretion such as glaucoma.
The effectiveness of the agents of the invention in inhibiting inflammatory disorders such as inflammatory airway diseases can be demonstrated in animal models, for example mouse or rat models of airway inflammation or other inflammatory disorders, e.g., as described in Szarka et al, J.Immunol.methods (1997)202: 49-57; renzi et al, am. Rev. Respir. Dis. (1993)148: 932-939; tsuyuki et al, J.Clin.1nvest, (1995)96: 2924-; and Cernadias et al, am.J.Respir.cell mol.biol. (1999)20: 1-8.
The substances of the invention may also be used as co-therapeutic agents for use in combination with other drug substances, such as anti-inflammatory, bronchodilatory or antihistamine drug substances, particularly for the treatment of obstructive or inflammatory airway diseases, such as those mentioned above, for example as potentiators of the therapeutic activity of these drugs or as a means of reducing the required dose of administration or potential side effects of these drugs. The agents of the invention may be mixed with the other drug substance in a fixed pharmaceutical composition or may be administered separately before, simultaneously with or after the other drug substance. The invention includes a combination of an agent of the invention as hereinbefore described with an anti-inflammatory, bronchodilatory or antihistamine drug substance, said agent of the invention and said drug substance may be in the same or different pharmaceutical compositions. Such anti-inflammatory agents include steroids, in particular glucocorticosteroids such as budesonide, beclomethasone dipropionate, fluticasone propionate, ciclesonide or mometasone furoate, and also WO 0200679, WO 0288167, WO 0212266 andcompounds described in WO 02100879, LTB4 antagonists such as those described in US5451700, LTD4 antagonists such as montelukast and zafirlukast, dopamine receptor agonists such as cabergoline, bromocriptine, ropinirole and 4-hydroxy 7- [2- [ [2- [ [3- (2-phenylethoxy) -propyl ] l]-sulfonyl group]-ethyl radical]-amino group]Ethyl radical]-2(3H) -benzothiazolone and its pharmaceutically acceptable salt (hydrochloride is
AstraZeneca), and PDE4 inhibitors such as
(GlaxoSmithKline), roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004(Bayer), SCH-351591(Schering-Plough), Alopeptine (AlmirallProdesfara), PD189659(Parke-Davis), AWD-12-281 (AstaMeica), CDC-801(Celgene), and KW-4490(Kyowa Hakko Kogyo), as well as those described in WO 98/18796 and WO 03/39544. Such bronchodilators include anticholinergics or antimuscarinics, particularly ipratropium, scopolamine bromoacetate and tiotropium salts, as well as those described in WO 01/04118, WO 02/51841, WO 02/53564, WO 03/00840, WO 03/87094, WO 04/05285, WO 02/00652, WO 03/53966, EP 424021, US 5171744, US 3714357 and WO 03/33495, as well as beta-2 adrenoceptor agonists such as albuterol, terbutaline, salmeterol and formoterol and pharmaceutically acceptable salts thereof, and the antihistamine drug substances for the combination treatment of compounds according to PCT International patent publication WO 00/75114 (which is incorporated herein by reference) of formula I (free form or salt form or solvate form), preferably embodiments thereof include cetirizine hydrochloride, Paracetamol, clemastine fumarate, promethazine, loratadine, desloratadine (desloratidine), diphenhydramine, and fexofenadine hydrochloride. Combinations of the agents of the invention with steroids, beta-2 agonists, PDE4 inhibitors or LTD4 antagonists are useful, for example, in the treatment of COPD or in particular asthma. Combinations of a substance of the invention with an anticholinergic or antimuscarinic agent, a PDE4 inhibitor, a dopamine receptor agonist or an LTB4 antagonist may be useful, for example, in the treatment of asthma or, in particular, COPD. Object of the inventionOther useful combinations of lipids with anti-inflammatory agents are combinations with antagonists of chemokine receptors, such as CCR-l, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9, CCR10, CXCRl, CXCR2, CXCR3, CXCR4, CXCR5, in particular CCR-5, such as Schering-Plough antagonists SC-351125, SCH-55700 and SCH-D, Takeda antagonists such as N- [ [4- [ [ [6, 7-dihydro-2- (4-methylphenyl) -5H-benzocyclohepten-8-yl ]]Hydroxy radical]Amino group]Phenyl radical]-methyl radical]tetrahydro-N, N-dimethyl-2H-pyran-4-ammonium chloride (TAK-770), and CCR-5 antagonists as described in US6166037 (especially claims 18 and 19), WO 00/66558 (especially claim 8) and WO 00/66559 (especially claim 9).
The substances of the invention may be administered by any suitable route, for example orally, such as in the form of tablets or capsules; parenteral, e.g., intravenous administration; administration by inhalation, for example in the treatment of inflammatory or obstructive airways diseases; intranasal administration, for example in the treatment of allergic rhinitis; topical application to the skin, for example in the treatment of atopic dermatitis; or rectal administration, for example in the treatment of inflammatory bowel disease.
The present invention also provides a pharmaceutical composition comprising a compound of the invention in free form or in pharmaceutically acceptable salt form, optionally together with a suitable pharmaceutically acceptable diluent or carrier. The composition may contain a combination therapeutic agent such as an anti-inflammatory, bronchodilator or antihistamine as described above. Such compositions may be prepared using conventional diluents or excipients and techniques known in the galenic art. Thus, oral dosage forms may include tablets and capsules.
Formulations for topical administration may take the form of creams, ointments, gels, or transdermal delivery systems such as patches. Compositions for inhalation may include aerosols or other aerosolizable or dry powder formulations.
When the composition comprises an aerosol formulation, it preferably contains, for example, a hydro-fluoro-alkane (HFA) propellant such as HFA134a or HFA227 or a mixture of these, may contain one or more co-solvents known in the art such as ethanol (up to 20% by weight), and/or one or more surfactants such as oleic acid or sorbitan trioleate, and/or one or more bulking agents such as lactose. When the composition comprises a dry powder it preferably contains, for example, a compound of the invention having a particle size of no more than 10 microns, optionally together with a diluent or carrier such as lactose having the desired particle size distribution, and a compound which helps to prevent deterioration of the product properties due to exposure to moisture. When the composition comprises a spray formulation, it preferably contains, for example, a compound of the invention dissolved or suspended in a medium containing water, a co-solvent such as ethanol or propylene glycol, and a stabiliser, which may be a surfactant.
The dosage of the agents of the invention used in the practice of the invention will vary depending upon, for example, the particular condition being treated, the desired effect and the mode of administration. Generally, suitable dosages for oral administration are on the order of 0.1 to 10 mg/kg.
In embodiments of the invention, where a patient is treated according to the invention, the amount of a given drug will depend on factors such as the particular dosing regimen, the type of disease or disorder and its severity, the uniqueness (e.g., body weight) of the subject or host in need of treatment, however, the dosage administered may be routinely determined by methods known in the art depending on the particular circumstances, including, for example, the particular drug that has been employed, the route of administration, the disorder being treated, and the subject or host being treated. In general, for dosages used for adult human therapy, dosages administered will typically range from 0.02 to 5000 mg/day, for example from about 1 to 1500 mg/day. The desired dose may conveniently be presented as a single dose, or as divided doses administered simultaneously (or over a short period of time) or at appropriate intervals, for example two, three, four or more divided doses per day. It will be appreciated by those skilled in the art that, notwithstanding the dosage ranges set forth above, the specific effective amounts may be adjusted as appropriate to the circumstances of the patient and in conjunction with the diagnosis by the physician.
Preparation of the Compounds
The compounds of formula (I) and formula (II) may be synthesized using standard synthetic techniques known to those skilled in the art or using methods known in the art in combination with the methods described herein. In addition, the solvents, temperatures, and other reaction conditions set forth herein may be varied according to the skill in the art. As a further guidance, the following synthesis methods may also be used.
The reactions may be used sequentially to provide the compounds described herein; or they may be used to synthesize fragments that are subsequently added by the methods described herein and/or known in the art.
In certain embodiments, provided herein are methods of making and methods of using PI3K and/or PfPI4K kinase inhibitor compounds described herein. In certain embodiments, the compounds described herein can be synthesized using the following synthetic schemes. The compounds can be synthesized using procedures analogous to those described below, by using appropriate alternative starting materials.
The starting materials for synthesizing the compounds described herein may be synthesized or may be obtained from commercial sources. The compounds described herein and other related compounds having different substituents can be synthesized using techniques and starting materials known to those skilled in the art. General methods of preparing the compounds disclosed herein can be derived from reactions known in the art, and the reactions can be modified by reagents and conditions deemed appropriate by those skilled in the art to incorporate various moieties in the molecules provided herein.
If desired, the reaction product may be isolated and purified using conventional techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like. These products can be characterized using conventional methods, including physical constants and profile data.
Non-limiting examples of compounds of the invention prepared are shown in table 1.
TABLE 1 Structure of example Compounds
Examples
The following specific non-limiting examples are to be construed as merely illustrative, and not limitative of the remainder of the disclosure in any way whatsoever. Without further elaboration, it is believed that one skilled in the art can, based on the description herein, utilize the present disclosure to its fullest extent.
Example 1: (S) - (1- ((4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) amino) -3-methyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester
(4- (5-amino-6-chloropyridin-3-yl) phenyl) carbamic acid tert-butyl ester (2): after 5-bromo-2-chloropyridin-3-amine (219 mg) was added to a round-bottom flask, 1, 4-dioxane (8 ml), water (4 ml), (4- ((tert-butoxycarbonyl) amino) phenyl) boronic acid (300 mg), Pd (PPh) were added 3 ) 4 (37 mg), potassium carbonate (300 mg). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (2), wherein LC/MS: m + H320.12.
(tert-butyl 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) carbamate (3): to a round bottom flask was added tert-butyl (4- (5-amino-6-chloropyridin-3-yl) phenyl) carbamate (240 mg) followed by pyridine (2 ml) and benzenesulfonyl chloride (0.57 ml). The reaction system is reacted for 14 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (3), wherein LC/MS: m + H460.11.
N- (5- (4-aminophenyl) -2-chloropyridin-3-yl) benzenesulfonamide (4): to a round bottom flask was added tert-butyl (4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) carbamate (100 mg) followed by ethyl acetate (4 ml) and methanol (1 ml) solution of hydrochloric acid. The reaction system was reacted at room temperature for 0.5 hour. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain compound (4), LC/MS: m + H360.06.
(S) - (1- ((4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) amino) -3-methyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester (5): after N- (5- (4-aminophenyl) -2-chloropyridin-3-yl) benzenesulfonamide (60 mg) was added to a round-bottomed flask, N, N-dimethylformamide (2 ml), (tert-butoxycarbonyl) -L-valine (38 mg), 2- (7-azobenzotriazol) -N, N, N ', N' -tetramethyluronium hexafluorophosphate HATU (78 mg), and N, N-diisopropylethylamine (0.2 ml) were added. The reaction was stirred at room temperature for 14 hours under argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (5), wherein LC/MS: m + H559.18.
Example 2: (S) -2-amino-N- (4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) -3-methylbutanamide
To a round bottom flask was added tert-butyl (S) - (1- ((4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) phenyl) amino) -3-methyl-1-oxobutan-2-yl) carbamate (50 mg) followed by ethyl acetate (2 ml) and methanol (1 ml) solution of hydrochloric acid. The reaction system was reacted at room temperature for 0.5 hour. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain compound (6) of example 2, LC/MS: m + H459.13.
Example 3: (S) - (1- (5- (6-chloro-5- (phenylsulfonamide) pyridin-3-yl) -1H-pyrrolo [2,3-b ] pyridin-1-yl) -3-methyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester
1- (5-bromo-1H-pyrrolo [2,3-b ] pyridin-1-yl) ethan-1-one (8): after 5-bromo-1H-pyrrolo [2,3-b ] pyridine (5.0 g) was added to a round-bottom flask, anhydrous dichloromethane (80 ml), triethylamine (3.6 ml) were added, and the system was cooled to 0 ℃ with an ice-water bath. Acetyl chloride was then added slowly. The reaction system is reacted for 14 hours at room temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with dichloromethane. The organic phase was washed with water and saturated brine, respectively, and then dried over anhydrous sodium sulfate. And filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a compound (8), wherein the mass ratio of LC/MS: m + H238.98.
1- (5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrrolo [2, 3-b)]Pyridin-1-yl) ethan-1-one (9): adding 1- (5-bromo-1H-pyrrolo [2,3-b ] into a round-bottom flask]Pyridin-1-yl) ethan-1-one (2.5g) was followed by addition of 1, 4-dioxane (80 mL), 4,4,4',4',5,5,5',5' -octamethyl-2, 2' -bis (1,3, 2-dioxaborane) (5.3g), PdCl 2 (dppf) 2 (257 mg) and potassium acetate (3.1 g). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 14 hours. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, evaporating to dryness under reduced pressureObtaining a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (9), wherein LC/MS: m + H287.16.
2-chloro-5- (1H-pyrrolo [2, 3-b)]Pyridin-5-yl) pyridin-3-amine (10): after 5-bromo-2-chloropyridin-3-amine (1.5g) was added to a round-bottomed flask, 1, 4-dioxane (80 ml), water (15 ml), 1- (5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1H-pyrrolo [2,3-b ] was added]Pyridin-1-yl) ethan-1-one (2.5g), Pd (PPh) 3 ) 4 (200 mg), potassium carbonate (2.5 g). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (10), wherein LC/MS: m + H245.06.
N- (2-chloro-5- (1H-pyrrolo [2,3-b ] pyridin-5-yl) pyridin-3-yl) benzenesulfonamide (11): to a round bottom flask was added 2-chloro-5- (1H-pyrrolo [2,3-b ] pyridin-5-yl) pyridin-3-amine (300 mg) followed by pyridine (8 ml) and benzenesulfonyl chloride (0.25 ml). The reaction system is reacted for 14 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressure silica gel column chromatography to obtain a compound (11), wherein LC/MS: m + H385.05.
(S) - (1- (5- (6-chloro-5- (phenylsulfonamide) pyridin-3-yl) -1H-pyrrolo [2,3-b ] pyridin-1-yl) -3-methyl-1-oxobutan-2-yl) carbamic acid tert-butyl ester (12): to a round-bottom flask was added N- (2-chloro-5- (1H-pyrrolo [2,3-b ] pyridin-5-yl) pyridin-3-yl) benzenesulfonamide (80 mg), followed by N, N-dimethylformamide (3 ml), (tert-butoxycarbonyl) -L-valine (48 mg), HATU (95 mg), and N, N-diisopropylethylamine (0.25 ml). The reaction was stirred at room temperature for 14 hours under argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (12), wherein the ratio of LC/MS: m + H584.18.
Example 4: 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -5, 6-dihydropyridine-1 (2H) -carboxylic acid tert-butyl ester
The synthesis of the compound of example 4 was accomplished using procedures analogous to those described in example 1. MS (ESI) M/z (M +1) + ：450.13。
Example 5: n- (2-chloro-5- (1,2,3, 6-tetrahydropyridin-4-yl) pyridin-3-yl) benzenesulfonamide hydrochloride
To a round bottom flask was added tert-butyl 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -5, 6-dihydropyridine-1 (2H) -carboxylate (100 mg) followed by ethyl acetate (3 ml) and methanol (1 ml) solution of hydrochloric acid. The reaction system was reacted at room temperature for 0.5 hour. After the reaction was complete, the system was evaporated to dryness under reduced pressure to afford the compound of example 5, LC/MS: m + H350.08.
Example 6: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylnicotinamide
5- (5-amino-6-chloropyridin-3-yl) -N-methylnicotinamide: to a round bottom flask was added 5-bromo-2-chloropyridin-3-amine (90 mg) followed by 1, 4-dioxane (4 ml), water (1 ml), N-methyl-5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinamide (135 mg), Pd (PPh) 3 ) 4 (16 mg), potassium carbonate (155 mg). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction is finished, the solvent is evaporated by the system under reduced pressure,the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain 5- (5-amino-6-chloropyridine-3-yl) -N-methylnicotinamide, and performing LC/MS: m + H263.07.
5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylnicotinamide: to a round bottom flask was added 5- (5-amino-6-chloropyridin-3-yl) -N-methylnicotinamide (41 mg) followed by pyridine (2 ml) and benzenesulfonyl chloride (0.15 ml). The reaction system is reacted for 14 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. The crude product was purified by pressure silica gel column chromatography to give the compound of example 6, LC/MS: m + H403.06.
Example 7: n- (5- (1-acetyl-1H-pyrrolo [2,3-b ] pyridin-5-yl) -2-chloropyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 7 was accomplished by using procedures analogous to those described in example 3. MS (ESI) M/z (M +1) + ：427.07。
Example 8: n- (2-chloro-5- (1-methyl-1H-pyrazol-4-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 8 was accomplished by using procedures analogous to those described in example 1. MS (ESI) M/z (M +1) + ：349.05。
Example 9: n- (2-chloro-5- (isoquinolin-4-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 9 was accomplished by using procedures analogous to those described in example 1. MS (ESI) M/z (M +1) + ：396.06。
Example 10: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N, N-dimethylnicotinamide
The synthesis of the compound of example 10 was accomplished by using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：417.08。
Example 11: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-cyclopropylnicotinamide
The synthesis of the compound of example 11 was accomplished using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：429.08。
Example 12: n-tert-butyl-5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamide
The synthesis of the compound of example 12 was accomplished using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：445.11。
Example 13: 5- (4-chloro-3- (phenylsulfonylamino) phenyl) -N-methylnicotinamide
N- (5-bromo-2-chlorophenyl) benzenesulfonamide (26): to a round bottom flask was added 5-bromo-2-chloroaniline (3.0g) followed by pyridine (15 ml) and benzenesulfonyl chloride (2.8 g). The reaction system is reacted for 14 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressure silica gel column chromatography to obtain a compound (26), wherein LC/MS: m + H345.93.
5- (4-chloro-3- (phenylsulfonylamino) phenyl) nicotinic acid (27): to a round-bottomed flask was added N- (5-bromo-2-chlorophenyl) benzenesulfonamide (690 mg), followed by 1, 4-dioxane (5 ml), water (5 ml), ethyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (500 mg), Pd (PPh) 3 ) 4 (200 mg), potassium carbonate (570 mg). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (27), LC/MS: m + H389.04.
5- (4-chloro-3- (phenylsulfonylamino) phenyl) -N-methylnicotinamide (28): after 5- (4-chloro-3- (phenylsulfinamido) phenyl) nicotinic acid (50 mg) was added to a round-bottomed flask, anhydrous tetrahydrofuran (2 ml), oxalyl chloride (140 mg), and N, N-dimethylformamide (1 drop) were added to the flask, and the reaction system was reacted at room temperature for 2 hours under an argon atmosphere. Then, methylamine (360 mg) was added to the system, and stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (28), LC/MS: m + H402.07.
Example 14: n-methyl-5- (6-methyl-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamide
N- (5-bromo-2-methylpyridin-3-yl) benzenesulfonamide (30): to a round bottom flask was added 5-bromo-2-methylpyridin-3-amine (1.5g) followed by pyridine (10 mL) and benzenesulfonyl chloride (1.56 g). The reaction system is reacted for 14 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated to dryness under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (30), wherein LC/MS: m + H326.99.
5- (6-methyl-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (31): to a round-bottomed flask was added N- (5-bromo-2-methylpyridin-3-yl) benzenesulfonamide (650 mg), followed by 1, 4-dioxane (5 ml), water (5 ml), methyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (500 mg), Pd (PPh) 3 ) 4 (200 mg), potassium carbonate (570 mg). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (31), wherein LC/MS: m + H370.09.
N-methyl-5- (6-methyl-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamide (32): after 5- (6-methyl-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (50 mg) was added to a round-bottom flask, anhydrous tetrahydrofuran (2 ml), oxalyl chloride (140 mg) and N, N-dimethylformamide (1 drop) were added to the flask, and the reaction system was reacted at room temperature for 2 hours under an argon atmosphere. Then, methylamine (360 mg) was added to the system, and stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (32), wherein LC/MS: m + H383.12.
Example 15: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -1H-pyrazolo [3,4-b ] pyridine-1-carboxylic acid tert-butyl ester
The synthesis of the compound of example 15 was accomplished by using procedures analogous to those described in example 3. MS (ESI) M/z (M +1) + ：486.10。
Example 16: n- (2-chloro-5- (1H-pyrazolo [3,4-b ] pyridin-5-yl) pyridin-3-yl) benzenesulfonamide hydrochloride
The synthesis of the compound of example 16 was accomplished by using procedures analogous to those described in example 15. MS (ESI) M/z (M +1) + ：386.05。
Example 17: 3- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylbenzamide
The synthesis of the compound of example 17 was accomplished using procedures analogous to those described in example 1. MS (ESI) M/z (M +1) + ：402.07。
Example 18: n- (2-chloro-5- (1H-pyrrolo [2,3-b ] pyridin-5-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 18 was accomplished by using procedures analogous to those described in example 7. MS (ESI) M/z (M +1) + ：385.05。
Example 19: 5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylnicotinamide
N- (5-bromo-2-fluoropyridin-3-yl) benzenesulfonamide (38): to a round bottom flask was added 5-bromo-2-fluoropyridin-3-amine (2.0g) followed by pyridine (10 ml) and benzenesulfonyl chloride (1.5 ml). The reaction system is reacted for 20 hours at normal temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (38), LC/MS: m + H330.96.
Ethyl 5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinate (39): in a round bottom flask was charged with N- (5-bromo-2-fluoropyridin-3-yl) benzenesulfonamide (331 mg) followed by 1, 4-dioxane (1 mL), 5- (ethoxycarbonyl) pyridin-3-ylboronic acid (332 mg), PdCl 2 (dppf) 2 (81.6 mg), potassium acetate (294 mg). The reaction system is reacted for 14 hours at 100 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (39), wherein LC/MS: m + H402.09.
5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (40): after ethyl 5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinate (100 mg) was added to a round-bottom flask, ethanol (4 ml) and water (1 ml) were added, and the reaction was reacted at room temperature for 5 hours under argon atmosphere. Then, sodium hydroxide (360 mg) was added to the system, and stirred at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (40), wherein LC/MS: m + H383.12.
5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylnicotinamide (41): to a round bottom flask was added 5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (40 mg) followed by anhydrous tetrahydrofuran (1 ml), methylamine hydrochloride (14 mg), HATU (76 mg), N-diisopropylethylamine (0.1 ml). The reaction system is reacted for 14 hours at room temperature under the protection of argon. The solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (41), wherein the ratio of LC/MS: m + H387.09.
Example 20: 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylbenzamide
The synthesis of the compound of example 20 was accomplished by using procedures analogous to those described in example 18. MS (ESI) M/z (M +1) + ：402.07。
Example 21: 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methylpyridineamide
The synthesis of the compound of example 21 was accomplished by using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：403.07。
Example 22: n- (2-chloro-5- (pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 22 was accomplished by using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：346.04。
Example 23: n- (2-chloro-5- (pyridin-4-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 23 was accomplished by using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：346.04。
Example 24: n- (2-chloro-5- (6-methoxypyridin-3-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 24 was accomplished using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：376.05。
Example 25: n- (5- (1-benzyl-1H-pyrazol-4-yl) -2-chloropyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 25 was accomplished by using procedures analogous to those described in example 8. MS (ESI) M/z (M +1) + ：425.09。
Example 26: n- (2-chloro-5- (2-methoxypyridin-3-yl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 26 was accomplished using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：376.05。
Example 27: n- (5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -2- (dimethylamino) acetamide
N- (5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) acetamide (54): to a round bottom flask was added N- (5-bromo-2-chloropyridin-3-yl) benzenesulfonamide (200 mg) followed by 1, 4-dioxane (5 ml), N- (5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-3-yl) acetamide (165 mg), PdCl 2 (dppf) 2 (47 mg) and potassium acetate (134 mg). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 14 hours. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (54), LC/MS: m + H403.07.
N- (5- (5-aminopyridin-3-yl) -2-chloropyridin-3-yl) benzenesulfonamide (55): to a round bottom flask was added N- (5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) acetamide (150 mg) followed by methanol (5 ml) and 6M hydrochloric acid solution (2 ml). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 6 hours. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain a solid compound (55). LC/MS: m + H361.05.
N- (5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -2- (dimethylamino) acetamide (56): to a round bottom flask was added N- (5- (5-aminopyridin-3-yl) -2-chloropyridin-3-yl) benzenesulfonamide (35 mg) followed by tetrahydrofuran (1 ml), 2- (dimethylamino) acetic acid (18 mg), HATU (67 mg) and N, N-diisopropylethylamine (0.1 ml). The reaction was stirred at room temperature for 14 hours under argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (56), LC/MS: m + H446.11.
Example 28: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-ethylnicotinamide
Ethyl 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinate (57): a round-bottomed flask was charged with N- (5-bromo-2-chloropyridin-3-yl) benzenesulfonamide (1.0g) followed by 1, 4-dioxane (10 mL), ethyl 5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) nicotinate (0.84g), PdCl 2 (dppf) 2 (0.23g) and potassium acetate (0.68 g). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 14 hours. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (57), LC/MS: m + H418.07.
5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (58): to a round bottom flask was added ethyl 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinate (1.2g) followed by ethanol (5 ml) and 1M sodium hydroxide solution (5 ml). The reaction system is heated to 80 ℃ under the protection of argon and reacted for 10 hours. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water (50 ml) and adjusted to pH 3 with 1M hydrochloric acid. The precipitate was filtered and dried to give compound (58), LC/MS: m + H390.03.
5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-ethylnicotinamide (59): to a round bottom flask was added 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinic acid (50 mg) followed by tetrahydrofuran (1 ml), ethylamine hydrochloride (21 mg), HATU (99 mg) and N, N-diisopropylethylamine (0.11 ml). The reaction system is reacted for 14 hours at room temperature under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (59), LC/MS: m + H417.08.
Example 29: 2- (3- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamido) Ethyl carbamic acid Tert-butyl ester
The synthesis of the compound of example 29 was accomplished by using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：532.14。
Example 30: (R) -2- (3- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamido) propylcarbamic acid tert-butyl ester
The synthesis of the compound of example 30 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：546.16。
Example 31: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N- (2-hydroxy-2-methylpropyl) nicotinamide
The synthesis of the compound of example 31 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：461.11。
Example 32: n- (2-aminoethyl) -5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamide hydrochloride
The synthesis of the compound of example 32 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：468.07。
Example 33: n- ((R) -1-Aminopropan-2-yl) -5- (6-chloro-5- (phenylsulphonylamino) pyridin-3-yl) nicotinamide hydrochloride
The synthesis of the compound of example 33 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：482.08。
Example 34: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-phenylnicotinamide
The synthesis of the compound of example 34 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：465.08。
Example 35: n-benzyl-5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) nicotinamide
The synthesis of the compound of example 35 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：479.10。
Example 36: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N- (4-fluorophenyl) nicotinamide
The synthesis of the compound of example 36 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：483.07。
Example 37: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N- (3-fluorophenyl) nicotinamide
The synthesis of the compound of example 37 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：483.07。
Example 38: 5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N- (pyridin-2-yl) nicotinamide
The synthesis of the compound of example 38 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：466.08。
Example 39: n- (5- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) acetamide
The synthesis of the compound of example 39 was accomplished by using procedures analogous to those described in example 27. MS (ESI) M/z (M +1) + ：403.07。
Example 40: 4- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -N-methyl-1H-pyrrole-2-carboxamide
The synthesis of the compound of example 40 was accomplished using procedures analogous to those described in example 28. MS (ESI) M/z (M +1) + ：391.07。
Example 41: 5- (5- (4- (tert-butyl) phenylsulfonylamino) -6-chloropyridin-3-yl) -N-methylnicotinamide
The synthesis of the compound of example 41 was accomplished by using procedures analogous to those described in example 6. MS (ESI) M/z (M +1) + ：459.13。
Example 42: n- (5- (4-chloro-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
N- (5- (4-chloro-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide (80): to a round bottom flask was added N- (5-bromo-2-chlorophenyl) benzenesulfonamide (63 mg), followed by 1, 4-dioxane (1 ml), N- (5- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) pyridin-3-yl) acetamide (50 mg), PdCl 2 (dppf) 2 (15 mg), potassium acetate (42 mg). The reaction system is reacted for 14 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (80), wherein LC/MS: m + H402.07.
Example 43: n- (5- (6-fluoro-5- (phenylsulfonylamino) pyridin-3-yl) acetamide
The synthesis of the compound of example 43 was accomplished by using procedures analogous to those described in example 39. MS (ESI) M/z (M +1) + ：387.09。
Example 44: n- (5- (6-methyl-5- (phenylsulfonylamino) pyridin-3-yl) acetamide
Synthesis of the Compound of example 44 by Using a catalyst analogous toThe procedure described in example 39 was completed. MS (ESI) M/z (M +1) + ：383.12。
Example 45: 5- (6-chloro-5- (4- (isopropyl) phenylsulfonylamino) pyridin-3-yl) -N-methylnicotinamide
The synthesis of the compound of example 45 was accomplished using procedures analogous to those described in example 41. MS (ESI) M/z (M +1) + ：445.11。
Example 46: n- (5- (4-fluoro-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 46 was accomplished by using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：386.10。
Example 47: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 47 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：382.12。
Example 48: n- (5- (3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 48 was accomplished by using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：368.11。
Example 49: n- (5- (6-chloro-5- (2, 6-difluorophenylsulfonylamino) pyridin-3-yl) acetamide
The synthesis of the compound of example 49 was accomplished using procedures analogous to those described in example 39. MS (ESI) M/z (M +1) + ：439.05。
Example 50: n- (5- (6-chloro-5- (2-fluorophenylsulphonamido) pyridin-3-yl) acetamide
The synthesis of the compound of example 50 was accomplished by using procedures analogous to those described in example 39. MS (ESI) M/z (M +1) + ：421.06。
Example 51: n- (5- (4-methoxy-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 51 was accomplished by using procedures analogous to those described in example 39. MS (ESI) M/z (M +1) +: 398.12.
Example 52: n- (5- (4-chloro-3- (2, 6-difluorophenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 52 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：438.05。
Example 53: n- (5- (4-chloro-3- (2-fluorophenyl sulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 53 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：420.06。
Example 54: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) propionamide
N- (5- (5-aminopyridin-3-yl) -2-methylphenyl) benzenesulfonamide (93): to a round bottom flask was added N- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide (1.7 g) followed by methanol (20 ml) and hydrochloric acid (20 ml). The reaction system is reacted for 4 hours at 80 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure to obtain a solid compound (93). LC/MS: m + H340.11.
N- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) propionamide (94): after N- (5- (5-aminopyridin-3-yl) -2-methylphenyl) benzenesulfonamide (30 mg) was added to a round-bottom flask, propionic anhydride (0.5 ml) was added. The reaction system is reacted for 14 hours at 85 ℃ under the protection of argon. After the reaction was completed, the solvent was evaporated under reduced pressure, and the resultant was diluted with water and extracted with ethyl acetate. The organic phase was washed with water and saturated brine, and dried over anhydrous sodium sulfate. Filtering the organic phase, and evaporating to dryness under reduced pressure to obtain a crude product. Purifying the crude product by pressurized silica gel column chromatography to obtain a compound (94), LC/MS: m + H396.14.
Example 55: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) pivaloyl amide
The synthesis of the compound of example 55 was accomplished by using procedures analogous to those described in example 54. MS (ESI) M/z (M +1) + ：424.17。
Example 56: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) isobutyramide
The synthesis of the compound of example 56 was accomplished by using procedures analogous to those described in example 54. MS (ESI) M/z (M +1) + ：410.16。
Example 57: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) butanamide
The synthesis of the compound of example 57 was accomplished using procedures analogous to those described in example 54. MS (ESI) M/z (M +1) + ：410.16。
Example 58: n- (5- (4-methyl-3- (phenylsulfonylamino) phenyl) pyridin-3-yl) pentanamide
The synthesis of the compound of example 58 was accomplished by using procedures analogous to those described in example 54. MS (ESI) M/z (M +1) + ：424.17。
Example 59: 2- (6-chloro-5- (phenylsulfonylamino) pyridin-3-yl) -1H-indole-1-carboxylic acid tert-butyl ester
The synthesis of the compound of example 59 was accomplished using procedures analogous to those described in example 4. MS (ESI) M/z (M +1) + ：484.11。
Example 60: n- (2-chloro-5- (1H-indol-2-yl) pyridin-3-yl) benzenesulfonamide hydrochloride
The synthesis of the compound of example 60 was accomplished by using procedures analogous to those described in example 5. MS (ESI) M/z (M +1) + ：384.06。
Example 61: n- (2-chloro-5- (4- (trifluoromethoxy) phenyl) pyridin-3-yl) benzenesulfonamide
The synthesis of the compound of example 61 was accomplished using procedures analogous to those described in example 1. MS (ESI) M/z (M +1) + ：429.03。
Example 62: n- (5- (4-methyl-3- (3-methyl-phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 62 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：396.14。
Example 63: n- (5- (4-methyl-3- (3-trifluoromethyl-phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 63 was accomplished by using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：450.11。
Example 64: n- (5- (4-methyl-3- (4-methyl-phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 64 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：396.14。
Example 65: n- (5- (4-methyl-3- (2-methyl-phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 65 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) +: 396.14.
example 66: n- (5- (3- (4-isopropyl-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 66 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：424.17。
Example 67: n- (5- (3- (3, 4-dichloro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 67 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：450.05。
Example 68: n- (5- (4-methyl-3- (2-trifluoromethyl-phenylsulfonylamino) phenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 68 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：450.11。
Example 69: n- (5- (3- (2, 4-difluoro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 69 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：418.11。
Example 70: n- (5- (3- (2, 3-dichloro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 70 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：450.05。
Example 71: n- (5- (3- (3-chloro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 71 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：416.09。
Example 72: n- (5- (3- (2, 5-dimethyl-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 72 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：410.16。
Example 73: n- (5- (3- (2-chloro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 73 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：416.09。
Example 74: n- (5- (3- (2, 4-dimethyl-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 74 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：410.16。
Example 75: n- (5- (3- (3, 4-difluoro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 75 was accomplished by using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：418.11。
Example 76: n- (5- (3- (4-chloro-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 76 was accomplished using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：416.09。
Example 77: n- (5- (3- (3, 4-dimethyl-phenylsulfonylamino) -4-methylphenyl) pyridin-3-yl) acetamide
The synthesis of the compound of example 77 was accomplished by using procedures analogous to those described in example 42. MS (ESI) M/z (M +1) + ：410.16。
Example 78
In vitro inhibitory Activity (enzyme Activity) assay
In vitro enzyme activity assays to determine the IC of compounds on PI3K family I kinases (PI3K alpha, PI3K beta, PI3K delta, PI3K gamma), type III kinases (VPS34), and PI4K family kinases (PfpI4K, PI4KB (human source)) 50 The value is obtained. Protein kinases PI3K α, PI3K δ, PI3K γ, VPS34, PI4KB were all purchased from Invitrogen (usa); protein kinase PI3K β was purchased from Sigma (usa); PfPI4K was expressed by this laboratory in the following manner. The Bac-to-Bac expression system (Thermo Fisher, USA) was used to construct the PfPI4K protein expression system. Recombinant baculovirus was first constructed according to the Bac-to-Bac expression system instructions, 1000mL of SF9 cells (purchased from ATC, USA) were cultured in large quantities, the constructed recombinant baculovirus was infected into SF9 cells, cell pellet was collected by centrifugation at 1000 rpm for 5 minutes after 72 hours, cells were lysed by sonication, and supernatant was collected at 12000 rpm for 10 minutes. First purified using his-beads, and then further purified using an FPLC protein purifier (AKTA, GE, USA). The three substrates PIP2: PS, PI and PI: PS were all purchased from Invitrogen (USA).
Protein kinase PI3K alpha 5.4. mu.L (final concentration of 0.16 ng/. mu.L), PI3K beta 5.4. mu.L (final concentration of 6 ng/. mu.L), PI3K delta 5.4. mu.L (final concentration of 1 ng/. mu.L), PI3K gamma 5.4. mu.L (final concentration of 5 ng/. mu.L), VPS 345.4. mu.L (final concentration of 1.2 ng/. mu.L), PI4KB 5.4. mu.L (final concentration of 5 ng/. mu.L), and PfPI4K 5.4. mu.L (final concentration of 5 ng/. mu.L) diluted to a certain concentration were each reacted with 1. mu.L of test drug compound diluted in a gradient for 1h at room temperature (final concentrations of 10. mu.M, 1. mu.M, 0.3. mu.M, 0.1. mu.M, 0.03. mu.01. mu.M, 0.003. mu.M, and 0.001. mu.M, respectively).
Adding 6 μ L of ATP and substrate PIP2: PS mixture into each reaction tube of PI3K family I type kinase (ATP final concentration in kinase PI3K α and PI3K β reaction system is 10 μ M, and kinase PI3K δ and PI are addedThe final concentration of ATP in the 3K gamma reaction system is 50 mu M, the final concentration of substrate PIP2: PS is 50 mu M), and the reaction is carried out for 1h at 37 ℃. The reaction buffer was 50mM HEPES (pH 7.5), 3mM MgCl 2 1mM EGTA, 100mM NaCl, and 0.03% CHAPS.
To each reaction tube of the above PI3K family III type kinase, 6. mu.L of ATP and the substrate PI: PS mixture (final concentrations of 50. mu.M and 100. mu.M, respectively) were added and reacted at 37 ℃ for 1 hour. The reaction buffer was 50mM Hepes (pH 7.5), 0.1% CHAPS, and 1mM EGTA.
To each reaction tube of the above PI4K family kinase, 6. mu.L of ATP and the substrate PI: PS mixture (final concentrations of 50. mu.M and 100. mu.M, respectively) were added and reacted at 37 ℃ for 1 hour. The reaction buffer was 20mM Tris (pH 7.5), 0.5mM EGTA and 0.4% Triton X-100.
mu.L of the kinase solution after the reaction was placed in a 384-well plate (Corning, USA), and 5. mu.L of ADP-Glo was added TM (Promega, USA) reagent, and reacting at room temperature for 40min to stop the kinase reaction and consume the rest ATP.
Add 10. mu.L of kinase assay reagent to convert ADP to ATP, detect newly synthesized ATP using a coupled luciferase/luciferin reaction, and calculate Graphpad 7.0IC using Envision read-out mapping 50 The values, experimental results are shown in table 2.
TABLE 2 measurement of in vitro inhibitory Activity (enzyme Activity) ("-" indicates no detection)
Table 2 follows.
| IC 50 (μM) | Example 6 | Example 47 |
| PIK3α | 0.20 | 0.35 |
| PIK3β | 0.31 | 5.41 |
| PIK3δ | 0.33 | >10 |
| PIK3γ | 0.22 | >10 |
| VPS34 | 0.20 | 4.14 |
| PfPI4K | 0.0015 | 0.0029 |
| PI4KB (human source) | 1.42 | >10 |
Example 79
Detection method of plasmodium falciparum GI50
Giemsa dye liquor (purchased from Sigma, USA) is diluted with double distilled water 1:20 for standby, and is prepared for use at the moment. The lysates contained 3 × SYBR Green I (from invitrogen, USA), 1.5% triton-x-100. Uninfected erythrocytes (isolated from normal human blood) were transferred to a 15ml centrifuge tube at 2000RPM for 5 minutes, the supernatant was removed, the pellet was slowly added to the upper layer of ficoll solution (beijing, solibao), centrifuged at 800g for 20 minutes, and the supernatant was removed. Leukocytes (isolated from normal human blood) and ficoll reagent were added to 10ml pbs, and erythrocytes were washed 2 times for 5 minutes at 2000 RPM. The supernatant was aspirated off leaving a precipitate for use. Mu.l of bottom cell pellet was aspirated from a culture dish for culturing infected erythrocytes, and dropped on a glass slide to prepare a blood sheet, which was stained with Giemsa stain for 19 minutes, and the stain was washed off with double-distilled water to count the infection rate. The infected red blood cell cultures were transferred to 15ml disposable centrifuge tubes. The culture supernatant was removed by rotating at 2000RPM for 5 minutes. And (3) uniformly mixing the infected erythrocyte sediment, adding uninfected erythrocytes, and adjusting the infection rate to 1.8-2.4%. The packed red blood cells were adjusted to 4.8-5% by adding medium (Gibcol, USA) to the infected red blood cells. The infected red blood cell suspension was added to a 96-well plate at 100. mu.l/well. The final concentrations of the drugs of 10, 3, 1, 0.3, 0.1, 0.03, 0.01, 0.003 and 0 μ M were added to each well in sequence, and 3 wells of each 96-well plate were placed to have the same volume of uninfected erythrocytes of 100 μ l. The candle jar culture method was performed for 72 hours. After 72 hours, the 96-well plate was shaken on a shaker for 14 seconds, and then 30. mu.l of lysis buffer was added and shaken for another 10 seconds. The mixture was placed on a shaker at room temperature for 10 minutes at 120 rpm. Envision reading, absorption wavelength of about 497nm, emission wavelength of about 520nm, GI calculated using Graphpad 7.0 50 The value is obtained.
TABLE 3
Claims (19)
1. A compound of formula (II) or a pharmaceutically acceptable salt thereof:
wherein:
x is selected from CH or N;
R 1 and R 2 At least one of which is H and the other is selected from H, C 1-6 Alkyl, halogen and C 1-6 A haloalkyl group;
R 3 selected from H, halogen, C 1-6 Alkyl and C 1-6 An alkoxy group;
R 4 selected from C optionally substituted by amino 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, phenylaminocarbonyl optionally substituted by halogen, phenyl C 1-6 Alkylaminocarbonyl and pyridylaminocarbonyl.
2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 1 And R 2 At least one of which is H and the other is selected from H, methyl, 2-propyl, fluoro, chloro and trifluoromethyl.
3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 3 Selected from chlorine, methyl and methoxy.
4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R 4 Selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, acetamido, dimethylaminoacetamido, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl and 2-pyridylaminocarbonyl.
5. A compound or pharmaceutically acceptable salt thereof, wherein said compound is a compound selected from the following table:
6. a pharmaceutical composition comprising a compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient, and optionally other therapeutic agents.
7. Use of a compound of formula (II) or a pharmaceutically acceptable salt thereof, for the preparation of an inhibitor of PfPI4K kinase against plasmodium falciparum:
wherein:
x is selected from CH or N;
R 1 and R 2 Are all H;
R 3 selected from halogen and C 1-6 An alkyl group;
R 4 is selected from C 1-6 Alkylaminocarbonyl and C 1-6 An alkanoylamino group.
8. The use according to claim 7, wherein said compound is a compound selected from the group consisting of:
9. use of a compound of formula (I) or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment, prevention or amelioration of malaria:
wherein:
x is selected from CH or N;
ring a is selected from the group consisting of pyridyl, pyrrolyl, pyrrolopyridyl, pyrazolyl, pyrazolopyridyl, tetrahydropyridinyl, phenyl and indolyl, said ring a having a nitrogen atom optionally substituted with an amino protecting group;
R 1 and R 2 Each independently selected from H, C 1-6 Alkyl and halogen;
R 3 selected from H, halogen, C 1-6 Alkyl and C 1-6 An alkoxy group;
R 4 selected from H, C 1-6 Alkyl radical, C 1-6 Alkanoyl radical, C 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, phenyl C 1-6 Alkyl, phenylaminocarbonyl, phenyl C 1-6 Alkylaminocarbonyl, pyridylaminocarbonyl and tert-butoxycarbonyl, optionally substituted with amino, tert-butoxycarbonyl substituted amino, hydroxy or halogen.
10. Use according to claim 9, wherein the a ring is selected from pyridin-3-yl, pyridin-4-yl, pyrrol-3-yl, pyrrolo [2,3-b ] pyridin-5-yl, pyrazol-4-yl, pyrazolo [3,4-b ] pyridin-5-yl, tetrahydropyridin-4-yl, phenyl and indol-2-yl, the nitrogen atom of the a ring being optionally substituted by tert-butyloxycarbonyl.
11. The use of claim 9, wherein R 1 And R 2 At least one of which is H.
12. The use of claim 9, wherein R 3 Selected from chlorine, methyl and methoxy.
13. The use of claim 9, wherein R 4 Is selected from C 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, phenylaminocarbonyl, phenyl C 1-6 Alkylaminocarbonyl and pyridylaminocarbonyl, the above substituents being optionally substituted by amino, tert-butoxycarbonyl substituted amino, hydroxy or halogen.
14. The use of claim 9, wherein R 4 Selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, acetamido, dimethylaminoacetamido, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl and 2-pyridylaminocarbonyl.
15. The use of claim 9, wherein the compound has the following structure of formula (II):
wherein:
x is selected from CH or N;
R 1 and R 2 At least one of which is H and the other is selected from H, C 1-6 Alkyl and halogen;
R 3 selected from H, halogen, C 1-6 Alkyl and C 1-6 An alkoxy group;
R 4 selected from H, C optionally substituted by amino 1-6 Alkylaminocarbonyl radical, C 1-6 Alkanoylamino group, C 1-6 Alkylamino radical C 1-6 Alkanoylamino, phenylaminocarbonyl optionally substituted by halogen, phenyl C 1-6 Alkylaminocarbonyl and pyridylaminocarbonyl.
16. The use of claim 15, wherein R 1 And R 2 At least one of which is H and the other is selected from H, methyl, 2-propyl and fluoro.
17. The use of claim 15, wherein R 3 Selected from chlorine, methyl and methoxy.
18. The use of claim 15, wherein R 4 Selected from the group consisting of methylaminocarbonyl, ethylaminocarbonyl, acetamido, dimethylaminoacetamido, phenylaminocarbonyl, fluorophenylaminocarbonyl, benzylaminocarbonyl and 2-pyridylaminocarbonyl.
19. The use according to claim 9, wherein the compound is a compound selected from the following table:
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